Immunocytochemical evidence for an afferent GABAergic neurotransmission in the guinea pig vestibular system. To implicate gamma-aminobutyric acid (GABA) as an afferent neurotransmitter (AN), the localization of GABA synthesizing and degradation enzymes; L-glutamate decarboxylase (GAD) and GABA transaminase (GABA-T) was investigated by light and electron microscopy immunocytochemistry in guinea pig vestibular cristae and ganglion cells (GC). GAD-like immunoreactivity was exclusively confined to the sensory hair cell (HC) cytoplasm, suggesting that GAD synthesizes GABA in the HC. GABA-T like immunoreactivity was found within HC, nerve calyces, nerve fibers, and GC, suggesting its participation in terminating transmitter action. These results demonstrate the existence of a GABAergic system in the guinea pig vestibule and strongly support GABA as a vestibular AN.


Studies on the cellular localization of biochemical responses to catecholamines in the brain. Studies were undertaken to determine the cellular localization of the cyclic adenosine monophosphate (cAMP) response of various forebrain regions to beta-adrenoceptor stimulation. Using brain slices, it was found that the gliotoxin, fluorocitrate (FC), which blocks metabolism selectively in glial cells, virtually abolished the cAMP response to beta-receptor stimulation whereas the neurotoxin, kainic acid (KA), was without effect. FC was confirmed by electrophysiological recording to be selective for glial cells in the brain slices. Similar results were found for these agents on in vivo brain cAMP responses to beta-receptor stimulation using a new microdialysis technique to measure in vivo responses. It is concluded that the cAMP response to beta-adrenoceptor stimulation in various regions of the forebrain occurs predominantly in glia. To determine if this could be correlated with a second biochemical response to beta-receptor stimulation, preliminary studies were undertaken on the localization of the immediate early gene, c-fos, produced in the brain after in vivo stimulation of beta-receptors. It was found that unlike the cAMP responses the c-fos response to beta-receptor stimulation occurs predominantly in neurons. The possible relationship of these two responses is discussed.


[Advantages of the new radiopharmaceutic agent, 99mTC-mercaptoacetyltriglycine (99mTc-MAG3), in the diagnosis of kidneydiseases. A comparative study with 131I-OIH, 99mTc-DTPA and 99mTc-DMSA]. The properties of 99mTc-mercaptoacetyltriglycine (MAG3), a new radiopharmaceutic for dynamic renal scintigraphy, were assessed in a series of 138 patients with different diseases of the uropoietic system. The physiological values of renographic curves were determined as well as the characteristics of MAG3 excretion, which closely resembled the values for 131I-OIH. On comparing the values of separate MAG3 clearance with standard radiopharmaceutics in 26 subjects examined, very good correlation coefficients (r) were obtained: 0.98, 0.96, and 0.92 for 131I-OIH, 99mTc-DMSA, and 99mTc-DTPA, respectively. Global MAG3 clearance reached 0.64 of the OIH value. The higher quality of scinfigraphic imaging than seen with the substances used so far, the favorable pharmacokinetic properties and the low radiation load (H(E) = 0.51 mSv in adults) of MAG3 determine the indication spheres in which it can best be used to advantage. Of the radiopharmaceutics used to date, MAG3 yields the best results. It can well substitute 123I-OIH, which is not readily available in our conditions, and can thus be expected to be widely used in clinical practice.


Ultrastructural relations between beta-adrenergic receptors and catecholaminergic neurons. We performed dual electron microscopic immunocytochemistry to determine the precise cellular relations between beta-adrenergic receptors (beta AR) and catecholaminergic terminals within adult rat brains. An antibody, beta AR404, against a peptide corresponding to the C-terminus of the hamster lung beta AR (beta 2 subtype) together with an anti-tyrosine hydroxylase (TH), a catecholaminergic marker, were used. Results show predominant labeling for beta AR404 within small astrocytic processes (beta-A). This is in sharp contrast to earlier results which showed neuronal labeling when using antibodies against the third intracellular loop of the receptor and of neurons-plus-astrocytes labeled using antibodies against the whole beta AR molecule. beta-A within visual cortex and nuclei of the solitary tracts frequently contacted blood vessel basement membrane and TH-immunoreactive terminals. TH-immunoreactive axons forming axo-axonic juxtapositions with non-TH terminals were also noted to be surrounded by beta-A. In the area postrema, a brain region lacking a blood-brain barrier, few beta-A occurred adjacent to TH-immunoreactive terminals or elsewhere. Thus, 1) catecholamines may act beyond morphologically identifiable synapses; 2) beta-A may mediate interactions between catecholamines and other transmitters; 3) there may be substantial heterogeneity in the structure or the conformation of the beta AR protein between neurons and glia or across CNS regions.


Nutritional and environmental factors involved in egg shell quality of laying hens. 1. Two experiments were conducted to assess the effects of lighting regime, dietary calcium source and sodium bicarbonate on production variables and egg shell quality of White Leghorn hens. In both experiments, hens were assigned to one of three lighting programmes that provided evening, morning or intermittent (midnight) lighting supplements in addition to natural daylight. Experimental diets used in the first study were formulated to contain (1) ground oyster shell flour, (2) limestone flour, or (3) and (4) the same +2/3 of the calcium source as hen-size oyster shell grit. The same 4 diets plus those containing hen-size limestone or hen-size limestone and oyster shells were used in experiment II. Additionally, diets in the first experiment contained either 0 or 10 g/kg sodium bicarbonate. 2. Hen-day egg production and food consumption were not affected by any of the experimental treatments. Hens fed on oyster shell diets or exposed to intermittent lighting regimes laid eggs of the highest specific gravity. Shell quality, as measured by specific gravity, was not affected by the addition of dietary sodium bicarbonate. 3. As expected, elevated temperatures (greater than 32 degrees C) significantly reduced egg shell quality. However, this effect was variable particularly in experiment II which used younger hens. 4. The shell quality of eggs from hens exposed to intermittent lighting in experiment II was significantly higher in each of the 4 sampling periods: morning (08.00-12.00), afternoon (12.00-16.00), evening (16.00-20.00) and night (20.00-08.00). 5. It is suggested that midnight lighting programmes provide a means of supporting egg shell quality of older laying hens during the summer months without a significant reduction in egg production.


Functional significance of cyclic AMP secretion in cerebral cortex. Cyclic AMP secretion in response to beta adrenergic receptor stimulation has been demonstrated in glioma-derived cell lines, in cerebral cortex in dissociated cell culture, and in the frontal cortex of living animals. The possible functions of cAMP secretion are considered; in particular, a role for this phenomenon in mediating some of the actions of norepinephrine as a neuromodulator in cerebral cortex.


Norepinephrine-mediated protein phosphorylation in astrocytes. Studies were conducted to determine if norepinephrine activates both protein kinase C and the cyclic AMP-dependent protein kinase in cultured rat astrocytes using phosphoproteins as markers. Norepinephrine was found to decrease 32P incorporation into an acidic 80,000 M(R) protein. A similar response was observed with isoproterenol and cyclic AMP analogs. In contrast, phorbol myristate acetate (PMA) increased 32P incorporation into this protein. Further studies looked at phosphorylation sites on glial fibrillary acidic protein and vimentin using two-dimensional tryptic phosphopeptide maps. The pattern of phosphorylation of these two proteins by norepinephrine resembles that of 8-bromo cyclic AMP and isoproterenol, and not that of PMA. Additionally, the effect of norepinephrine on the phosphorylation of GFAP and vimentin was blocked by alprenolol. One difference noted between norepinephrine and isoproterenol was the phosphorylation of an 18,000 M(R) protein. Norepinephrine increased, and isoproterenol decreased, 32P incorporation into this protein; however, the mechanism which mediates the norepinephrine effect remains to be determined. Overall, these studies indicate that the most prominent phosphorylation events mediated by norepinephrine are the consequence of the activation of cyclic AMP-dependent protein kinase.


Excess sodium bicarbonate in the diet and its effect on Leghorn chickens. 1. A commercial 62-week-old layer flock experienced an acute drop in egg production and an increase in shell-less egg production within 2 days of consuming feed erroneously formulated to contain over 30 g/kg instead of 3 g/kg sodium bicarbonate (NaHCO3). Other symptoms included increased water consumption, diarrhoea and increased mortality associated with visceral gout. 2. An experiment was conducted to assess the responses of hens under controlled conditions. Twenty Dekalb XL Single Comb White Leghorn hens (50 weeks old) were placed in individual cages, having ad libitum access to water from trough waterers. Ten hens were fed the TEST (High NaHCO3) feed for one week (Test group), and ten hens remained on normal commercial layer ration (Control group). 3. Hens in the Test group had high water consumption and watery droppings, but egg production and mortality were not affected. Physiological evaluations indicated the Test feed caused metabolic alkalosis. Plasma sodium, urine pH and urinary sodium excretion were increased, and glomerular filtration rates were decreased in the Test group. 4. These physiological effects are consistent with known responses to excess sodium intake in domestic fowl. The reduced egg production and increased mortality caused by the Test feed under commercial conditions may be related to more severe dehydration experienced by hens in multi-bird cages supplied by cup-type watering systems.


Effect of selective opiate antagonists on striatal acetylcholine and dopamine release. We investigated the effect of selective opiate antagonists on striatal acetylcholine (ACh) and dopamine (DA) release. The mu-receptor antagonist beta-funaltrexamine (beta-FNA), the delta-antagonist naltrindole (NTI), and the kappa-antagonist norbinaltorphimine (nor-BNI) were used to selectively block different subtypes of opiate receptors. The experiments were carried out on isolated superfused striatal slices of rats, loaded with [3H]choline or [3H]dopamine. beta-FNA and NTI significantly enhanced the electrical field stimulation-evoked release of ACh but only if the dopaminergic input had been impaired either by chemical denervation or D2 dopamine receptor blockade. By contrast, neither the selective nor nonselective antagonists had any modulatory effect on the release of dopamine. It is concluded, therefore, that the release of ACh is tonically controlled by endogenous opioid peptide(s) through the stimulation of mu- and delta-opiate receptors located on cholinergic axon terminals, in addition to the tonic control by DA.


Preserved neurotransmitter receptor binding following ischemia in preconditioned gerbil brain. Preconditioning the brain with sublethal ischemia induces tolerance to subsequent ischemic insult. Using [3H]quinuclidinyl benzilate (QNB), [3H]MK 801, [3H]cyclohexyladenosine, [3H]muscimol, and [3H]PN200-110, we investigated the alterations in neurotransmitter receptor and calcium channel binding in the gerbil hippocampus following ischemia with or without preconditioning. Two-minute forebrain ischemia, which produced no neuronal damage, resulted in no alterations in binding except for a slight reduction in [3H]QNB binding in the CA1 subfield. Three-minute ischemia destroyed the majority of CA1 pyramidal cells and caused, in CA1, reductions in binding of all ligands used. Preconditioning with 2-min ischemia followed by 4 days of reperfusion protected against CA1 neuronal damage and prevented the reductions in binding although [3H]QNB and [3H]PN200-110 binding transiently decreased in the early reperfusion period, suggesting down-regulation. Thus, preconditioning protects against damage to the neurotransmission system as well as histopathological neuronal death.


Arterial catheterization.

Arterial catheterization. Arterial catheterization is used frequently in the management of critically ill patients, both for continuous blood pressure monitoring and access to the arterial circulation to obtain frequent blood gas measurements. The procedure is usually easily accomplished at the bedside using percutaneous methods such as the Seldinger technique to cannulate the radial, brachial, axillary, femoral, or dorsalis pedis artery. Meticulous attention to aseptic technique is necessary during insertion and catheter maintenance to minimize the risk of catheter-related infection. Other potential complications include hemorrhage, ischemia, arteriovenous fistula, and pseudoaneurysm formation.


Pericardiocentesis.

Pericardiocentesis. Cardiac tamponade is a life-threatening condition resulting from compression of the cardiac chambers by a pericardial effusion. The principal cause of pericardial effusion is malignant disease of the pericardium, but infectious causes and cardiac trauma are common as well. The patient with cardiac tamponade demonstrates an abnormal pulsus paradoxus, and clinical signs of shock and impending cardiovascular collapse occur with very severe cardiac compression. Relief of the increased intrapericardial pressure is mandatory to establish adequate cardiac output. The definitive treatment of cardiac tamponade is emergent removal of enough pericardial fluid to acutely lower intrapericardial pressure. Echocardiographic guidance may be used if immediately available, but is not required to perform pericardiocentesis in a critical situation. Placement of a pulmonary artery catheter prior to pericardiocentesis is not indicated in cardiac tamponade. Once cardiac output and tissue perfusion have been restored, further drainage procedures such as pericardial catheter placement or surgical drainage are indicated. Therapeutic measures to address the underlying disease process should be initiated after pericardial drainage is accomplished.


[Surveillance of fetal growth and fetal cerebellar transverse diameter by ultrasonographic measurement]. Measured fetal transverse cerebellar diameters were obtained by ultrasonography in 325 women during 20-42 weeks of gestation. A nomogram for transverse cerebellar diameter with respect to gestational age was generated. Curvilinear relationship was found between the transverse cerebellar diameter and the gestational age (R = 0.99624, P less than 0.0005). Fetal transverse cerebellar diameter, biparietal diameter, head circumference, abdominal diameter, abdominal circumference and femur length were measured by ultrasound in 39 women not more than one week before delivery. Correlation coefficients between the birth weights and these parameters were studied. It was found that the correlation between the transverse cerebellar diameter and the birth weight was the highest. The function of the transverse cerebellar diameter in the evaluation of fetal growth and development is better than any other parameter. The transverse cerebellar diameters of 20 IUGR cases were obtained. It was indicated that the growth of the transverse cerebellar diameter slowed down in primary symmetric IUGR, and was unaffected in secondary asymmetric IUGR. Thus it has clinical diagnostic value in symmetric IUGR. When combined with the abdominal circumference, the fetal cerebellar transverse diameter may help to differentiate the types of growth-retarded fetuses.


[Antihypertensive effects of highly active atrial natriuretic peptide (haANP) infusion in patients with pregnancy-induced hypertension]. To study the antihypertensive effects of a new synthetic highly active atrial natriuretic peptide (haANP), the effects were observed in patients with moderate and severe pregnancy-induced hypertension (PIH). There were significant decreases in blood pressure (BP) for 10 min after haANP infusion (from 20.46 +/- 1.04/14.15 +/- 1.59 kPa to 18.17 +/- 0.60/11.08 +/- 1.36 kpa; P less than 0.05). 90 min after haANP infusion, BP decrease was the lowest (to 16.00 +/- 1.21/10.5 +/- 0.71 kPa; P less than 0.01), while plasma ANP levels increased 11 folds (from 47.5 +/- 5.5 ng/L to 525.4 +/- 15.6 ng/L). The effects continued for 300 minutes. Antihypertensive effects of magnesium sulphate were relatively relaxed (compared with those of haANP). The results showed strong and rapid antihypertensive effects of haANP. We found that mean RA and ALD levels were significantly decreased after haANP.


[Regularity of recurrent sites of cervix carcinoma after radical operation]. Radical operations were performed in 872 cases of cervical carcinoma from April 1964 to December 1985. The concrete ways of operation and auxiliary therapy were different according to clinical stage of each case. All the cases were followed-up for more than 5 years. 114 cases showed definite single or multiple lesions. Analysis of 92 cases of single lesions, the revealed that recurrent sites may be related to clinical stage, pathology, pelvic node metastases and auxiliary treatment. The local recurrence rate of single lesions was 59.8% and that of distant lesions was 40.2%.


[Intrathyroidal T cell activation and HLA-DR antigen expression on thyroid follicular cells in autoimmune thyroid diseases]. Thyroid specimens from 19 patients with Hashimoto's thyroiditis (HT), 11 with Graves' disease (GD), 4 with nontoxic goiter (NTG), 1 with subacute thyroiditis (SAT), 1 with thyroid adenoma and 4 from normal thyroids were investigated by alkaline phosphatase anti-alkaline phosphatase (APAAP) immunocytochemical technique. A group of monoclonal antibodies against the corresponding T cell activation antigens were used. The positive rates of all the four activation antigens in thyroid gland mononuclear cells (TG-MNC) were significantly higher in HT than in NTG (P less than 0.05-0.01). However, the differences between HT and GD were insignificant (P greater than 0.05) except for HLA-DR antigen. The activation antigen-positive (especially TLiSA 1+) TG-MNC were often seen intruding into thyroid lumens of HT. All the abnormal specimens expressed HLA-DR antigens on thyroid follicular cells (TFC) in different degrees (+/- to +3), and the degree in HT was significantly higher than that in GD (P less than 0.01) or NTG (P less than 0.05). The level of DR expression on TFC correlated significantly with the infiltrating degrees of T-activation-antigen-positive cells (P less than 0.01). This indicates that aberrant DR expression in vivo is closely related to the activation of intrathyroidal T cells.


[Model of atrophic gastritis in rats by various method]. Model of atrophic gastritis was made in rats by synthetical method which comprised twice of active immunization and continual oral administration of bile and hot water for 70 days. There was a highly significant difference (P less than 0.01) between the normal group and testing group in the number of rats with changes in body-weight, PH value of gastric mucous membrane, gastric membrane blood flow (GMBF) and its potential difference (PD), and pathology (thinned mucous membrane, decreased gland, infiltration of inflammatory cells and intestinal metaplasia). By observations for additional 45 days, the above-mentioned values still showed significant or highly significant differences (P less than 0.05 or P less than 0.01) between the testing group and the normal group. The animal model of atrophic gastritis made by this method can be used for the valuation of drug efficacy.


[Protective effects of ginsenosides on warm ischemic damages of the rabbit kidney]. Renal function and morphology were studied before and after 60 min of renal ischemia and contralateral nephrectomy in two groups of rabbits. The animals were pretreated with ginsenosides (n = 22) and saline (n = 22) respectively, the latter as control. Results showed that ginsenosides (30 mg/kg body wt.) pretreatment by intravenous injection 10 min before warm ischemia resulted in the survival of all the animals with better renal function, 1, 3 and 7 days after blood urea nitrogen, fraction of excreted sodium and urine protein were observed in the control rabbits and a less pronounced increase was noted (P less than 0.05) after pretreatment with ginsenosides. The appearance of kidney tissue taken from surviving rabbits with Ginsenosides pretreatment was found to be normal under light microscope. Severe tubular necrosis was observed in kidneys of the control group. Tissues were examined with a transmission electron microscope. ginsenosides have protective effects on the epithelial cells of the proximal convoluted tubules, and microvilli and mitochondria were less damaged by ischemia than those of the control animals. There was also a large amount of ribosome on rough surfaced endoplasmic reticulum in the cells of ginsenosides-treated kidney, reflecting their ability to stimulate ribonucleic acid and protein synthesis. This is considered to be the basis of improvement of renal function.


[Immunohistochemical study of transferrin receptor expression in hepatocellular carcinoma]. By means of immunohistochemical technique ABC, using monoclonal anti-transferrin receptor (TFR) antibodies WuT9 and OKT9, TFR expression in 30 cases of hepatocellular carcinoma (HCC) and in 6 cases of organs and tissues of normal human bodies was studied. It was revealed that large amount of TFR were expressed in liver cancer cells, but not in the surrounding mesenchymal cells as demonstrated by intense immunostaining in cancer nests, and even not in the surrounding mesenchyma of those HCC patients with negative AFP in their serum. In normal human body, only small amount of TFR in limited sites was found without free antigen in blood stream. Thus, it followed that TFR as a structural antigen of HCC was expressed with higher relative specificity than AFP, and TFR may be considered a tumor marker and therapeutic target of HCC.


[Epidemiological study of selective IgA deficiency among 6 nationalities in China]. The frequency of selective IgA deficiency (SIgAD) was determined in 33,171 Chinese distributed in six nationalities by Ouchterlony's double diffusion with 5 mg/dl as the limit of detection. Those who were found IgA-deficient were retested by single radial diffusion technique. Gel diffusion analysis of sera revealed apparent lack of IgA in 8 samples, all with normal levels of IgG and IgM (age-matched). The incidence was 0.024% (1:4,100). The incidences among the six nationalities were Han 1:2,600, Hui 1:5,000, Mongolia 1:2,700, Tujia 1:4,300, Zhuang 0:5,300, and Bai 1:4,800. All of the individuals, who had low levels of IgA or no IgA detectable, were below 16 years of age. Of these, 2 sera were autoantibody (anti-smooth muscle antibody) positive Delayed hypersensitivity skin tests were negative in one individual. The chief clinical manifestations were those of respiratory tract infections, of which about 50% were in subclinical state.


[Changes in T cell subsets and the expression of receptors of transferrin and interleukin-2 in chronic active hepatitis]. The changes of T-cell subsets and expression of transferrin receptor (TFR) and interleukin-2 receptor (IL 2 R) of peripheral blood lymphocyte from patients with chronic active hepatitis (ACH) and chronic serious hepatitis (CSH) were studied. The results showed that T3+ cells were decreased. (P less than 0.01) and T8+ cells were significantly increased (P less than 0.01) in patients with CAH and CSH. TH/TS ratio was markedly decreased in both groups of patients (P less than 0.01). The data also showed that TFR and IL 2 R were in a state of high-expression in resting T-cell, but no obvious changes could be found in the expression of receptor of PHA-stimulated T cells of patients, as compared the controls. The results suggested that some inhibitory factor may be present in the serum of CAH and CSH patients.


[Mechanism of the photosensitive effects of methylene blue in the inhibition of DNA synthesis in cancer cells]. In the present study, we found that various concentrations of methylene blue (38, 3.8, 0.38, 0.038, 0.0038 mumol/L) significantly inhibited DNA synthesis in ascites hepatoma (AH) cells. The inhibition increased with the concentration of methylene blue as well as with irradiation time. Photosensitization of methylene blue was not due to 1O2, OH., or O2.-, but was closely related to H2O2. The mechanism of inhibition of DNA synthesis may be attributed to the damage of DNA replication template.


[Interleukin 1, transforming growth factor beta and messenger ribonucleic acid expression during the course of glomerulonephritis and glomerulosclerosis]. The accumulation of extracellular matrix is a prominent feature of progressive glomerulonephritis and glomerulosclerosis. This study was designed to evaluate the coeffective role of interleukin 1 (IL-1), transforming growth factor beta (TGF beta), and accumulation of collagen IV, fibronectin (FN), laminin (LN) in the progress of glomerulonephritis. Rat mesangial proliferation and sclerosis was induced by injection of anti-thymocyte serum (ATS). Total RNA of glomeruli were extracted from rat kidney at 3, 7, 14, 28, 60 days after ATS administration for northern blots. The results showed that IL-1, TGF beta mRNA expression was 2 to 3 folds higher than in the controls on the 7th to 14th day after injection of ATS. At the same time, there were increased accumulations of collagen IV, FN, and LN. Increased IL-1, TGF beta mRNA expression and accumulation of collagen IV, FN, LN were closely associated with mesangial proliferation, matrix expansion and focal glomerulosclerosis. These results suggest that increased IL-1, TGF beta mRNA expression and accumulation of collagen IV, FN, LN may play an important role in the progression of glomerulonephritis.


[Cytomegalovirus-DNA in serum and renal tissue of patients with IgA nephropathy]. Cytomegalovirus (CMV) has been suspected to be involved in the pathogenesis of IgA nephropathy (IgAN). In this study, IgAN both with and without recurrent gross hematuria (GH), cases of MsPGN, MN were included, 10 cases in each group. Sera were obtained and analyzed for the presence of CMV-DNA with polymerase chain reaction. Renal biopsies of the 20 cases of IgAN were also analyzed for the presence of CMV-DNA with in situ hybridization. The positive rates of serum CMV-DNA were 7/10 in IgAN (GH), 2/10 in IgAN (non-GH), 1/10 in MsPGN and 1/10 in MN. It was much higher in IgAN (GH) than in the other groups. There was only 1 case which showed CMV-DNA in renal tissue in IgAN (GH) and IgAN (non-GH) respectively. The high frequency of CMV-DNA present in the serum but not in the renal tissue of IgAN implicated that there was a close association but not etiological relationship between them.


[HLA-DR gene frequencies in IgA nephropathy patients obtained by oligonucleotide genotyping]. HLA-DR alleles of 255 healthy northern Chinese donors and 30 IgA nephropathy (IgAN) patients were determined by using a set of 30 different sequence specific oligonucleotide (SSO) probes directed to various DRB alleles. We found that SSO typing gave high gene frequencies for the alleles DR2, DR7 and DR 9 in the northern Chinese Han donors, while we obtained significantly high gene frequency for DRw12 in IgAN, especially in those with massive proteinuria and recurrent gross hematuria, indicating that those who have DRw12 may form a high risk population of IgAN.


[Gene deletion of X-linked ichthyosis].

[Gene deletion of X-linked ichthyosis]. The characterization of steroid sulfatase (STS) gene mutation from seven X-linked ichthyotic patients was performed by multiple polymerase chain reaction (MPCR) which amplified two specific regions at the 5' and 3' end of STS gene. The results indicated that five out seven patients were found to have entire STS gene deletion. Two other cases and five patients' mothers showed two amplification fragments. So did two cases of dominant ichthyosis vulgaris. It was further ascertained that entire gene deletion is the main mutation of STS locus in Chinese population. MPCR is a rapid and simple method for gene diagnosis of X-linked ichthyosis.


[Characterization of monoclonal antibody (BIU-H) directed against bladder cancer and its pathologic feature]. The characteristics of one of a group of monoclonal antibodies named BIU-H6 are reported. Using McAb BIU-H6, by ABC-immunohistochemical staining directed against 34 bladder cancers and 5 normal bladders. It was found that BIU-H6 does not react with normal bladder epithelial and adenocarcinoma of the bladder, but with transitional cell carcinoma, squamous cell carcinoma and CIS of the bladder. The staining characters were variant according to the different grade/stage of the cancer. The result shows that BIU-H6 antigen is a tumor associated antigen of poorly differentiated and invasive carcinoma of the bladder.


[Immunogold dot assay for diagnosis of early pregnancy]. Two monoclonal antibodies anti-HCG were prepared. One anti-alpha HCG was directly labeled with colloidal gold, and the other, anti-beta HCG was coated on nitrocellulose membrane as dot. After determination of optimal conditions of the system, an immunogold dot blot assay for testing HCG in urine sample was established. The sensitivity of the assay was 50 mIU HCG/ml, corresponding to the content of HCG in the urine of women about 30 days after last menstruation. The assay has been evaluated in clinical trial. All the testing results accorded with clinical diagnosis. No false positive results appeared. This simple, rapid and stable assay is ideal for women to diagnose early pregnancy at home.


[Expression of hepatitis B virus X protein in tumor and nontumor tissues of patients with hepatocellular carcinoma (HCC)]. PAP technique and rabbit anti-X serum were used to detect the X protein in tumor and nontumor liver tissues from 34 patients with HCC. The positive rate of the X protein in both tissues were 94.1% and 84.4% respectively. Of the 34 patients with HCC, 27 were complicated by liver cirrhosis, in whom 92.6% were X protein positive in liver cells. It was found that almost all of the liver cells adjacent to the tumor tissue showed strong positive staining. The high frequency and predominant expression of X protein in HCC and liver cirrhosis tissues indicated that X protein may play an important role in hepatocarcinogenesis. X protein was detected in 17.2% of the patients with CAH, which suggested the risk of transformation from CAH to cirrhosis and/or HCC. X protein was first found in bile duct epithelial cells in 59.4% of the patients with HCC, and 6 of 34 HCC were combined with bile duct carcinoma, and some cancer cells were found positive for X protein. It seems that X protein may also be a potential factor in the oncogenesis of bile duct carcinoma.


[Changes in plasma glucagon concentration in patients with diabetes mellitus and its clinical significance]. To investigate the role of glucagon in the pathogenesis of diabetes, we observed change of plasma glucagon concentrations during glucose loading in normal subjects and patients with impaired glucose tolerance (IGT) and non-insulin dependent diabetes mellitus (NIDDM) using specific radioimmunoassay. The results showed that the fasting plasma glucagon levels in patients with IGT and NIDDM were similar to those of the normal subjects. The nadir of plasma glucagon level in the normal control occurred at 1-h after glucose loading and the changes of glucagon, glucose and insulin levels synchronized; but peaks of plasma glucose and insulin levels in the IGT and NIDDM patients were delayed at 2-h after glucose loading with the lowest glucagon level at 3-h. It was suggested that there were relative hyperglucagonemia and decrease of sensitivity of islet A cell to glucose in IGT and NIDDM patients. The present study also showed that hyperglucagonemia is related to the reduction of insulin secretion and might play an important role in the development of postprandial hyperglycemia in NIDDM.


[Immunocytochemical study of CuZn superoxide dismutase in the myocardium of normal subjects and patients of rheumatic heart disease]. Using the methods of immunocytochemistry and immune electron microscopy, we studied the subcellular localization of GuZn superoxide dismutase (CuZn SOD) in the myocardium of 8 normal subjects and the changes of CuZn SOD of the myocardium in 51 patients of rheumatic heart disease, and at same time, we also determined the CuZn SOD contents in the myocardium with radioimmunoassay. This study demonstrated that most normal myocardial cells contained CuZn SOD. The ventricular myocardial cells stained somewhat stronger than the atrial myocardial cells. CuZn SOD was mainly distributed in the cytoplasm and the nucleus, less in mitochondria, a little in lysosomes and almost no gold particle was seen on the membranous structures of sarcoplasmic reticula and Golgi's complex. The positive level of CuZn SOD in rheumatic myocardium had remarkable correlation with myocardial fibrosis and arteriopathy and the recovery of the heart functions after valve replacement. The changes of CuZn SOD in various organelles in the myocardium of patients with rheumatic heart disease may be possibly related to the injuries of their membranous structure.


[Retinoic acid receptor alpha gene rearrangement as specific marker of acute promyelocytic leukemia and its use in the study of cell differentiation]. The chromosomal translocation t (15; 17), hallmark of human acute promyelocytic leukemia (APL), has been shown to disrupt the retinoic acid receptor alpha (RARA) gene. Using a panel of probes covering the whole RARA gene generated in our own laboratory, we detected the gene rearrangement in 23 out of 25 cases of APL. It is conceivable that RARA gene rearrangement may serve as a highly specific marker of APL. The RARA gene configuration was investigated in two APL patients during the course of treatment with all-trans retinoic acid (ATRA). It was shown that phenotypically differentiated bone marrow myelocytes under ATRA therapy could still carry RARA gene rearrangement. This provides further evidence that ATRA induce complete remission of APL through differentiation induction of the leukemic cells.


[Mechanical principles of L-trapezoid compression plate in the fixation of trochanter fractures]. Three pairs of human cadaver femur were used for simulation electric test with load on both legs and single leg. An Evans III osteotomy was made, and stabilized with L-trapezoid compression plate (L-TCP). L-angled plate (L-AP) 95 degrees and 130 degrees respectively. The results demonstrated that the stability of fixation of L-TCP was significantly greater than L-AP. Since the cancellous screw of L-TCP can hold the proximal fragment and forms a stable triangular truss with the L end, the bony support of the medial posterior part can be reconstructed by holding the lesser trochanter with the lag screw. From June 1980 to August 1990, 68 patients with 71 trochanter fractures and osteotomies were treated with L-CTP. 54 patients with 57 fractures and osteotomies were followed up for 6-52 months. All fractures and osteotomies were healed. No bone cutting off failure was encountered. The hips and knees of 51 patients (54 limbs) (94.8%) showed excellent and good functions. Infection occurred in 3 patients (4.4%). Varus deformity developed in 2 patients with an Evans IV fracture, and plate fracture occurred in one of them.


[The effects of early stage load-bearing on bone ingrowth of porous coated implant in rabbits]. To understand the effects of load-bearing on cementless fixation of porous coated implant, we inserted Co-Cr-Mo alloy implants into the distal femur of rabbits. Specimens were studied by means of histology, anti-collagen type I and III immunohistochemical staining, colloidal gold labeled immunoelectron microscopy, and computerized graphic analysis. The results revealed that stimulations of load-bearing benefited bone ingrowth, but the newly formed bone in the pores was absorbed and replaced by fibrous tissues after 12 weeks without stimulations of load-bearing. The fibrous tissues in the pores of the implant were primarily type III collagen after one week and type I collagen after two weeks. After 6 weeks, the residual tissue between ingrowth bone and metal bound antibodies was type I collagen. Fibroblasts participated in and possessed the action of osteogenesis.


[Experimental reconstruction on intrinsic hand muscle function by anterior interosseous nerve transference]. The anterior interosseous nerve was transferred to the recurrent branch of the median nerve and the deep branch of the ulnar nerve respectively to restore the function of the intrinsic hand muscles. Twelve Rhesus monkeys were used with 24 nerves for transference and 24 for direct suturing. Gross, histological, and ultrastructural observations, electrophysiological evaluation and computer-imaging analysis system were employed to assess the nerve regeneration process in 1,3,6, and 12 months after operation. The results showed that the rate of maturity and the number of regenerating nerves in the distal segment approached the normal level and the passing rate of the myelinated axons reached to 156.16 +/- 14.45% 12 months after operation. According to the histological findings and the recovery of the intrinsic hand muscle function, reinnervation in the transferred nerves occurred 1 to 3 months earlier than that in the directly sutured ones.


[Iatrogenic injuries of the peripheral nerves: analysis of 226 cases]. In 226 patients with iatrogenic peripheral nerve injuries, the causes were analyzed in detail. The iatrogenic nerve injuries were incurred through injection of therapeutic agents in 84 patients, inadvertent division by the surgeon in 44, traction during orthopaedic operation in 27, and undue compression from an ill-fitting splint or plaster cast in 26. The prophylaxis of such iatrogenic lesions is discussed. It cannot be over-stressed to exercise special care to prevent the nerve from being injured in the course of dissection or in any therapeutic maneuver, and to grasp a precise knowledge of pertinent anatomy on the part of the surgeon. Iatrogenic nerve injury is avoidable and, once incurred, satisfactory results can be expected and its consequences minimized when recognized and treated early and properly.


[Erythrocyte membrane band 3 protein and HCO3-/Cl- exchange function in cor pulmonale patients]. The changes of erythrocyte membrane band 3 protein, blood gases and electrolytes of intraerythrocyte and extra-erythrocyte were investigated in 3 groups: type I respiratory failure (group I, n = 36), type II respiratory failure (group II, n = 33) and control group (CG, n = 50). The distribution of band 3 protein was narrow and the staining intensity of band 3 protein was lower in the electrophotogram of group II. The relative composition of band 3 protein in group II was significantly lower than that in group I and CG (P less than 0.01). The intraerythrocyte HCO3- in group II was significantly higher than that in group I and CG (P less than 0.01), but the extra-erythrocyte Cl- in group II was significantly lower than that in group I and CG (P less than 0.01). These findings suggested that (1) The relative composition reduction of erythrocyte membrane band 3 protein and HCO3-/Cl- exchange restrain may be one of the reasons that aggravated CO2 retention and respiratory acidosis in cor pulmonale patients with type II respiratory failure. (2) Because there was hypochloremia in the most cor pulmonale patients with type II respiratory failure, it was necessary to supply them enough chloride in time, which could not only correct hypochloremia, but also accelerate the rate of HCO3-/Cl- exchange and promote to eliminate CO2.


[Dynamic study on superficial blood flow in patients with acute myocardial infarction]. Fourteen points superficial bloodflow (SBF) of the skin and tongue in 55 patients with acute myocardial infarction (AMI) were measured at 12, 24, 36, 48, 60 hours and 3, 5, 7, 10, 14, 21, 28 days after AMI by LDF (PF2). The dynamic study on multiple points SBF of 576 times showed that (1) The mean SBF of skin was 1.0 +/- 0.04 (V) at 12 hours after AMI. It occupied 70.5% in the normal control. After therapy at 48 hours of AMI, the mean SBF was increased to 1.20 +/- 0.03 (V), and approximated 85.9% of the control. (2) The mean SBF in patients with cardiogenic shock was 1.04 +/- 0.05 (V), and it was significantly lower than that without complications (P less than 0.01). The mean SBF showed a negative correlation with the nailfold microcirculatory values (P less than 0.0025). There was a negative correlation between "Tanzhong" SBF and cardiac muscle enzyme CPK, GOT, LDH (P less than 0.05). (3) The SBF of acupuncture point "Tanzhong, Erxin" related to the heart might more sensitively represent the cardiac condition in AMI. (4) The SBF of tongue was negatively correlated with GOT (P less than 0.05). (5) Continual peripheral microcirculatory observation and electrocardiographic monitoring would be helpful in the diagnosis and treatment of earlier complications of AMI in order to reduce the mortality.


[Experimental studies on the antitumor activity of glioma-infiltrating lymphocytes against autologous glioma]. In the present study, antitumor activity of glioma-infiltrating lymphocytes (GILs) was compared to that of lymphokine-activated killer (LAK) cells. Results suggested that killing activity of GILs against autologous glioma cells was significantly higher than that of LAK cells (P less than 0.05), but their activity against allogeneic glioma cells was not different from that of LAK cells (P greater than 0.05). Analysis of cell surface phenotypes showed that CD4+ cells were a main portion of LAK cells, and CD8+ were predominant in the GILs. In the beginning, growth of GILs was slower than that of LAK cells. As time went on, generation of GILs was faster than that of LAK cells. The results suggested that GILs were superior to LAK cells for adoptive immunotherapy in patients with brain glioma.


Quality of life assessment in end-stage renal disease patients with maintenance hemodialysis therapy. This is a pilot study assessing the quality of life for a total of twenty uremic patients being treated with maintenance hemodialysis. It includes eight males and twelve females with a mean age of fifty three years, having been on hemodialysis for an average of forty-five months (from five to 143 months). The quality of life study was obtained from a structured questionnaire, which contained 10 categories including: sense of well-being, sense of mood, family life, marriage life, neighborhood relationships, friend relationships, working ability, job condition, intelligence condition and life satisfaction. Each category consisted of 3 to 5 items, with a maximum score of 100 points. Among these 20 patients, the highest score on the quality of life study is marriage life with 80.4 +/- 10.0 points, the lowest score is life satisfaction with 59.7 +/- 15.3 points. There was no significant difference in the scores of quality of life between different sex groups. The scores in the sense of well-being group greater than 60 years were lower than those less than 60 years with a significant difference (P less than 0.05). A significant difference was also found in both the sense of well-being and sense of mood depending on the duration of hemodialysis therapy. Those who received hemodialysis therapy more than 3 years produced better scores in the sense of well-being and sense of mood category than those who received hemodialysis less than 3 years.


Ca2+ uptake by endoplasmic reticulum of renal cortex. I. Ionic requirements and regulation in vitro. A subcellular fraction enriched in cytochrome c reductase (7.9-fold) and relatively de-enriched (0.64-fold) in Na+/K(+)-ATPase was prepared from canine kidney cortex by sucrose density gradient ultracentrifugation. It was shown by electron microscopy to consist primarily of a light fraction of endoplasmic reticulum (LER). LER vesicles displayed ATP-dependent 45Ca2+ uptake that was insensitive to 10 mM KCN or NaN3, and was promptly released by 20 microM A23187 or ionomycin. Inositol-1,4,5-trisphosphate (IP3) appeared to produce a time-dependent release of 45Ca2+. Vanadate inhibited 45Ca2+ uptake with a Ki approximately 0.3 mM, further suggesting that the activity resided in the ER rather than the plasma membrane. 45Ca2+ uptake by LER, at 5 microM total [Ca2+], displayed a strong dependence on divalent cations (Mg2+ greater than Co2+ greater than Mn2+ much greater than Ba2+ greater than or equal to Cd2+ greater than or equal to Sr2+, present at 2 mM) as well as on monovalent cations (Na+ greater than or equal to K+ + Na+ greater than K+ greater than Li+ greater than choline +), and anions (Cl- greater than acetate- greater than or equal to NO3- greater than or equal to F- greater than H2PO4- much greater than gluconate- greater than or equal to oxalate= much greater than SO4=). It had a fairly narrow pH optimum (7.25-7.50). Preincubation (10 min) of LER vesicles with 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated LER Ca2+ uptake; this effect was enhanced in the presence of renal cytosol [5% (vol/vol)].(ABSTRACT TRUNCATED AT 250 WORDS)


Shoulder computed arthrotomography: role of internal and external rotation. Computed arthrotomography of the shoulder is the best investigation for instability of the glenohumeral joint. The purpose of this paper is to evaluate the role of internal rotation and external rotation in detecting glenoid and capsular lesions. From October 1989 to December 1991, 74 double-contrast CT arthrograms of the shoulder were performed with both internal and external rotation. Most of these patients were referred for instability problems or shoulder pain of vague origin. Of the 49 abnormalities of the anterior portion of glenoid labrum, 44 (89.8%) were identified on the scans with internal rotation; however, 5 (10.2%) was seen only in external rotation view. Most of the anterior capsular abnormalities occurred on the internal rotation. Only one of 74 examination was evidenced solely on external rotation. There were relatively few posterior labral or capsule abnormalities. Of the five, two were seen solely on external rotation. The results suggest that internal rotation scans detect most of the lesions; however, with the help of external rotation view, 10% increase in diagnostic yield can be expected for anterior labral abnormalities. Although few posterior labral or capsule lesions occur, external rotation examination does play an important role in this respect.


A new autoperfusion technique for aortic reconstruction of suprarenal aortic aneurysm. Visceral ischemia is a serious factor in the postoperative morbidity and mortality of suprarenal aortic reconstruction. We reported two patients of suprarenal aortic aneurysm involving visceral arteries, who received successful Dacron graft replacement by using Pruitt-Inahara balloon catheters as an autoperfusion for preservation of the visceral organs. No visceral organ ischemia occurred postoperatively except in patient 2 who had preoperative chronic renal failure and persistent renal failure after the operation. The renal function recovered gradually during the follow-up period. Both patients are doing well at the present time. The new autoperfusion technique can directly deliver normothermic blood from the arterial cannula at proximal aorta to the individual visceral arteries by using the balloon perfusion catheters. It is simple, safe, easily instituted and the used products are readily obtainable. It allows the surgeon to provide an effective protection of visceral organs for the suprarenal aortic reconstruction.


Ultrasonically guided needle aspiration biopsy in the diagnosis of advanced superior vena cava syndrome. From July 1990, 11 cases with advanced superior vena cava (SVC) syndrome were undergone ultrasonically guided needle aspiration or biopsy. Ten cases were diagnosed by thin-needle aspiration cytology. Two cases with cytologic proven lymphoma and one case without cytologic diagnosis were undergone needle biopsy, and two of them were proven to be lymphoma. The diagnostic rate of aspiration cytology was 91.0% (10/11) and of aspiration biopsy was 66.6% (2/3). The diagnostic rate of combination of aspiration cytology and biopsy was 100%. For reducing the incidence of pneumothorax, we strongly recommend this two-stage procedure that needle biopsy was performed only when cytologic diagnosis was not conclusive. Only one case developed pneumothorax in performing needle biopsy. In conclusion, ultrasonically guided needle aspiration biopsy is a safe, convenient, and high-yield diagnostic procedure for selected cases of advanced SVC syndrome.


Roentgenographic determination of total lung capacity in normal Chinese children. Roentgenographic determination of total lung capacity (TLC) in children, rarely mentioned in the past, differs from the determination of TLC using helium dilution method in that it does not need the measurement of functional residual capacity. Therefore, it is useful for hospitals where pulmonary function testing (PFT) is not available. In this study, 87 out of 125 normal children completed both chest radiographs and PFT. Ellipsoid method which divides the lung field into several sections was used to calculate TLC from chest radiographs. The correlation coefficient of TLC measured by both chest radiographs and PFT was 0.7680, and paired Student's t-test of the mean values of the two groups showed P value greater than 0.05. Among the variables of height, weight, age and body surface area, height alone was able to predict the normal reference value of TLC. Line of regression between TLC and height was expressed as "TLC = 52.776 x Height- 4404.998", and correlation coefficient was 0.8013. Intersubject reproducibility was also verified. It is concluded that chest radiographs can be used to determine TLC in children.


[Latent structure model of physicians' opinions on National Health Insurance]. The purpose of this study is to construct the latent structure models of physicians' opinions on National Health Insurance (NHI). The data for the analysis came from a mail survey of 1619 physicians in January of 1990. Five latent structure models were established as follows: Structure 1. Physicians' viewpoint on NHI. Hospital physicians: Steady, 69%; Turn-Over, 26%; Career-change, 5%. Clinic physicians: Steady-with Insurance Contract, 55%; Steady-without Insurance Contract, 41%; Career-Change, 4%. Structure 2. Physicians' expected impact of NHI. Hospital physicians: Pessimistic, 45%; Disadvantaged, 38%; Constant, 17%. Clinic physicians: Pessimistic, 72%; Constant, 12%; Ambitious, 16%. Structure 3. Physicians' expected workload change due to the implement of NHI. Hospital physicians: Decreasing, 30%; No-Change, 18%; Increasing, 52%. Clinic physicians: Decreasing, 30%; Non-Change, 23%; Increasing, 48%. Structure 4. Ideal practice pattern. Hospital physicians: Traditional, 47%; Transitional, 42%; Practice Abandoned, 12%. Clinic physicians: Traditional, 20%; Transitional, 21%; Rural-Orientated 59%. Structure 5. Expected payment methods for physicians. Hospital physicians: Credentiality-Specialty-based, 44%; Specialty-Equal Pay-based, 11%; Equal Pay-Specialty-based, 42%; Equal Pay-Credentiality-based, 4%. Clinic physicians: Credentiality-based, 24%; Mixed, 11%; Equal Service-Equal Pay-based, 60%; Urbanization Level-based, 5%.


[Acute abdomen with intraperitoneal bowel perforation-demonstration by CT scan]. Ninety-eight cases of surgically proved bowel perforation in VGH-Kaohsiung within one year were reviewed. Among them, 25 patients received abdominal CT scans before operation. Free gas or fluid could be demonstrated with CT scan in 80% of cases though sometimes the fluid was too scanty to be well recognized. Local abscess formation with or without tumor mass lesion (n = 3), local fluid accumulation (n = 1) and negative CT findings (n = 1) constituted the remaining 20% of cases. The recognition of perforation sites depended on direct evidences of perforation such as ulceration, abscess formation and tumor mass lesion (32%), or indirect evidences including hematoma formation, dependent extraluminal gas accumulation and phlegmonous reaction (12%). An inexplicable localized extraluminal fluid accumulation might also suggest a possibility of hollow organ perforation and lead to the recognition of perforation site in the absence of free gas (4%). Absence of extraluminal fluid or gas can not completely rule out the possibility of hollow organ perforation (4%). Massive free air, especially with an air-fluid level, suggested a possibility of upper abdominal perforation. The perforation hole located at the anterior wall of stomach produced more free gas than that located at posterior wall. Air-fluid levels could never be identified in the cases of lower gastrointestinal perforations. Local bowel wall thickening and the distribution of free gas contributed little to the recognition of perforation site. Extravasation of oral contrast media sometimes led to the diagnosis of gastrointestinal perforation but was not necessary to be noted around the perforation hole and did little contribution to the perforation site recognition.(ABSTRACT TRUNCATED AT 250 WORDS)


[Second hip fracture].

[Second hip fracture]. During the period from 1982-1990, a total of 1644 patients were admitted to our hospital with proximal femoral fractures. Trochanteric fractures were encountered in 54 per cent of the cases (887/1644) with a mean age of 68 years. Cervical fractures were encountered in 46 per cent of the cases (757/1644) with a mean age of 62 years. We found 28 patients who had been readmitted with a second hip fracture in this retrospective study, including twenty three males and five females. Two of these second hip fractures occurred on the same side as the initial fracture and twenty six occurred on the opposite side. The time interval between the first and the second hip fracture was shorter in males than that in females (1.7 years V.S. 2.7 years) and the interval also was shorter in those with a previous trochanteric fracture as compared to those with a previous cervical fracture (1.5 years V.S. 2.6 years). It appeared that a hip fracture reduced the risk of a subsequent hip fracture on the same side, and almost all second hip fractures were located on the opposite side of the patient (93 per cent). A possible reason for this "protective mechanism" is increased strength of the bone due to callus formation. A significant symmetry (trochanteric-trochanteric or cervical-cervical) was found between the first and the second hip fractures (69 per cent). We believe the symmetrical fracture pattern is due to senile osteoporosis, a kind of ongoing systemic weakening of the skeletal system among the elderly.


[Projected supply and geographic distribution of dentists in Taiwan for the year 2000]. Using the data from the Dentist Masterfile of the Department of Health, Executive Yuan, and the reports from all Colleges of Medicine in Taiwan and the Ministry of Defense, this study projected the future supply of dentists as well as their geographic distribution. The components of projection included studying current supply, future increment and loss. The results of this study showed that there would be 7897 active dentists in Taiwan by the year 2000, or 36 dentists per 100,000 population. It also showed the poor geographical distribution of dentists will not substantially ameliorate in response the rapidly increasing supply of dentists. The 14 to 1 ratio of the highest to the lowest (Taipei city V.S. Penghu county) in terms of dentist-population will remain in the coming decade. Finally, this study discussed the projection process as well as the implications for dentist manpower policy in Taiwan.


Adrenal hemangioma: computed tomogram and angiogram appearances. Adrenal hemangiomas are rare. To our knowledge, about 22 cases have been reported in the literature, of which 13 cases were surgically removed. We report probably the first case of CT and angiographically diagnosed and surgically confirmed adrenal hemangioma in Taiwan. We concluded that characteristic appearances on computed tomogram and angiogram associated with phlebolith-like calcification in the tumor may allow the radiologists to make correct preoperative diagnosis.


Solitary metastasis of renal cell carcinoma to the ovaries: a case report. Though renal cell carcinoma may metastasize to unusual sites via hematogenous spread, ovarian metastasis is very uncommon. This is explained by atrophy of the ovaries with decreased blood perfusion in most postmenopausal patients who are at the age of peak incidence of renal cell carcinoma. We report a 28-year-old woman with metastatic renal cell carcinoma to the ovaries. Special attention to the differential diagnosis and the management should be taken.


Acute cerebellitis with ocular flutter and truncal ataxia: a case report. This report entails a case of acute cerebellitis manifested with ocular flutter and truncal ataxia on a 24-year-old man. He became dizzy and unsteady beginning October 7, 1991. Transient horizontal to-and-fro movement of objects while gazing, and prominent gastrointestinal tract symptoms were also present. A neurological examination showed ocular flutter and truncal ataxia. Cerebrospinal fluid studies showed mild lymphocytic pleocytosis. Computed tomogram and magnetic resonance imaging of the head were normal. Electronystagmography showed typical ocular flutter in various eye positions. The ocular flutter and truncal ataxia subsided gradually and disappeared completely after October 28th.


[Percutaneous ultrasound-guided thick catheter drainage of retroperitoneal abscess: report of 2 cases]. Retroperitoneal abscess have been a difficult diagnostic and therapeutic challenge in the past. With the advent of precise, noninvasive imaging modalities, they now are defined more easily. Percutaneous guided catheter drainage has changed our traditional therapeutic approach to this disease, in the majority of cases. In general, massive semilytic fatty tissue of retroperitoneal abscess can not pass through the ordinary percutaneous catheter. Herein we report 2 cases of retroperitoneal abscess successfully treated with percutaneous guided thick catheter drainage combined with medical control. Percutaneous guided thick catheter drainage can be performed in most uroradiological facilities. There are many advantages to percutaneous ultrasound-guided thick catheter drainage compared with surgical drainage of retroperitoneal abscess. They include: earlier diagnosis and treatment, avoidance of general anaesthesia, operations which involve risks and complications, shorter and less expensive therapy, easier nursing care, better infection control with the closed drainage system, and greater patient acceptance. Therefore, we believe that a trial of thick catheter percutaneous drainage of retroperitoneal abscess, without a complex septum, should be the initial mode of therapy, and surgical intervention should be undertaken only when percutaneous drainage fails or is contraindicated.


Long-term follow-up of hepatitis B vaccination in infants born to noncarrier mothers in Taiwan. Two hundred and seventy-seven infants born to hepatitis B surface antigen (HBsAg)-negative noncarrier mothers had been divided into 3 groups to receive 1 microgram, 2 micrograms or 5 micrograms of plasma derived hepatitis B vaccine and were followed up annually to the age of 6. At the end of the follow-up period, the antibody to HBsAg (anti-HBs) positive rates in these 3 groups were 63.0% (58/92), 77.9% (74/95) and 83.3% (75/90), respectively (group 1 vs. group 2, p less than 0.05; group 1 vs. group 3, p less than 0.01; group 2 vs. group 3, p greater than 0.05). Among the initial responders, 25 of 38 (65.8%) with low initial anti-HBs titers (10-100mIU/ml) and 18 of 79 (22.8%) with medium initial anti-HBs titers (101-1,000mIU/ml) were found to be anti-HBs seronegative, while only 2 of 136 (1.5%) with high initial anti-HBs titers (greater than 1,000 mIU/ml) lost their anti-HBs (P less than 0.01). None of the vaccinees became HBsAg positive but 1 initial responder became positive for anti-HBc at the age of 4. Twenty-five initial nonresponders and 18 initial responders who lost their anti-HBs at the age of 2 received a second booster of vaccination at the age of 3. Five (20.0%) of the initial nonresponders and 17 (94.4%) of the initial responders responded to the second booster. The annual incidence of natural booster was 2.1%. In conclusion, the standard doses of 5 micrograms plasma-derived vaccine tend to be the best choice for immunization of infants born to noncarrier mothers in Taiwan.(ABSTRACT TRUNCATED AT 250 WORDS)


The establishment of radioimmunoassay of cyclic AMP. Succinyl cyclic AMP, coupled with human albumin, was injected into rabbit to elicit antibodies to the cyclic nucleotide hapten. The succinyl cyclic nucleotides were conjugated to human albumin through a carbodiimide coupling procedure. The radioligand was prepared essentially by the modified method of Hunter and Greenwood. The resulting radiolabeled 125-ScAMP-TME was subsequently purified by reverse phase chromatography on Seppak C18 cartridge and used as tracers. Free and bound form of labeled antigens in the assay system were separated according to their differences in adsorption to solid material, i.e., charcoal. The antisera in routine use had 8% cross-reactivity with cyclic GMP, but had no cross-reactivity with ADP, 5'-AMP or ATP. The sensitivity of the assay in standard was around 0.05 pmol/ml. The sequential saturation analysis system was superior to the equilibrium study in terms of its sensitivity in standard curve determination. The media dilution and recovery studies showed good parallelism with the standard curve. The intra-assay and inter-assay coefficient variations were 5% and 7%, respectively. It showed that a significant increase in tracer binding to the antibodies in equilibrium incubating system was obtained when they were incubated at 4 degrees C for up to 72 hours. When the standard curve was derived for Scatchard plot analysis, Kd of the curve decreased inversely with the increase of the antibodies diluted, the total binding capacity of the antibody would not alter. A thyroid-stimulating hormone produced a normal response of cyclic AMP concentrations in FRTL-5 tissue cultures.


Epithelial neoplasm of the colorectum: correlation of histopathological features with cytogenetic studies. Seven cases of primary rectosigmoid epithelial neoplasm, including a tubular adenoma, a villous adenoma with malignant change and five adenocarcinomas, were cytogenetically studied with short-term cultivation and GTG-banding. The tubular adenoma showed karyotypic normality. The other six malignant tumors were hyperdiploid in chromosomal modal number and showed multiple aberrations. The complexity of chromosomal aberrations increased proportionally to the pathologic stage. No common chromosome change to all of the tumors was found. The most common recurrent chromosome changes were extra gains of chromosomes #7 and #8q, which were found respectively in four of the six malignant tumors. Two recurrent structurally abnormal chromosomes were i(8q) and der(10)t(10;?)(p15;?). No case showed sex chromosome change, except one tumor from a male patient who obtained an extra X chromosome. All these chromosome changes found were consistent with the results reported by other investigators. Deletions of #5q and #17 were nevertheless only found in one and two of our six malignant cases, respectively. No cases showed a deletion of #18q. The possible molecular mechanisms of these recurrent chromosome aberrations were also discussed.


Mutational analysis of genes determining antagonism of Alcaligenes sp. strain MFA1 against the phytopathogenic fungus Fusarium oxysporum. Alcaligenes sp. strain MFA1 inhibits microconidial germination and germination-tube elongation of Fusarium oxysporum f.sp. dianthi and reduces the severity of fusarium wilt of carnation, presumably as a result of its production of a siderophore (G.Y. Yuen and M.N. Schroth. 1986. Phytopathology, 76:171-176). Derivative strains of MFA1, deficient in antagonism against F. oxysporum and in iron-limited growth, were obtained by Tn5 mutagenesis. The presence of a single Tn5 insertion in the genomic DNA of each derivative strain was detected by Southern analysis. Marker-exchange mutagenesis of strain MFA1 with DNA fragments, containing Tn5 and flanking sequences cloned from representative mutants, confirmed the association of single Tn5 insertions with the loss of antifungal activity and iron-independent growth of MFA1. These results are consistent with the involvement of siderophore biosynthesis by MFA1 in the inhibition of F. oxysporum.


Chromosome mobilization of Legionella pneumophila with RK2::Mu and Tn5-Mob. RK2::Mu plasmids and transposon Tn5-Mob were used to mobilize the Legionella pneumophila chromosome. Plate matings between L. pneumophila donors that contained RK2::Mu plasmids and auxotrophic recipients yielded recombinants at frequencies ranging from 10(-6) to 10(-7) per recipient for the markers tested. The presence of a Mu insertion in the chromosome of donors that harbored RK2::Mu plasmids increased the frequency of chromosome transfer of certain selected markers as compared with strains that contained RK2::Mu alone. Cotransfer experiments with Mu-containing donors and a thymidine and tryptophan auxotroph failed to reveal any linkage between the thy and trp loci in L. pneumophila. A strain that contained a chromosomal Tn5-Mob insertion and helper plasmid pRK24.4 transferred chromosomal markers at frequencies of 10(-7) per recipient. These findings suggest that RK2::Mu plasmids and Tn5-Mob may be useful for genetic mapping experiments with L. pneumophila.


Antitumor effect of deferoxamine on human hepatocellular carcinoma growing in athymic nude mice. Iron is essential for the growth of all living cells. One of the important intracellular roles for iron is in the activation of ribonucleotide reductase, the enzyme that catalyzes the first step in DNA synthesis. Thus, the intracellular iron level may serve as a regulator of cell growth. The authors tested the hypothesis that lowering body iron concentration inhibits the growth of human-derived hepatocellular carcinoma (HCC) cells by depleting these cells of iron. Deferoxamine (DFO), an iron-chelating agent, was used to lower intracellular iron level. HCC cells, PLC/PRF/5 (7 x 10(6) cells/mouse), were transplanted subcutaneously into athymic nude mice. When tumors reached 200-300 microliters in size, mice with comparable tumor sizes were paired; one was treated with DFO (300 mg/kg body weight/day, 5 days/week) intraperitoneally while the other received no treatment. Eight pairs of mice with HCC were observed for 5-18 weeks. Mean tumor growth rates (TGR) (mean +/- standard error) for the untreated and treated mice were 30.5 +/- 3.7 microliters/week and 11.9 +/- 1.5 microliters/week. The difference was significant (P < 0.02). In the second set of studies, DFO treatment was begun when the tumor size was smaller (100-200 microliters). Four pairs of mice were observed for 4-15 weeks; mean TGR for the four untreated mice was 18.1 +/- 5.1 microliters/week. In two mice treated with DFO, tumors regressed completely by the seventh week after initiation of treatment. The two remaining mice on DFO therapy had much slower growing tumors, with a mean TGR of 1.8 +/- 0.5 microliters/week. Thus, our results suggest that (1) reduction of intracellular iron concentration by DFO may be useful as antitumor therapy in HCC and (2) the favorable effects of DFO treatment are best seen when treatment is begun when the tumor is small.


Lymphatic flow in carcinoma of the head of the pancreas. The lymphatic pathway from the head of the pancreas to the para-aortic lymph nodes was examined on the basis of the frequency of lymph node involvements. Forty-four patients were examined. All patients had extended radical operations. Thirty-one of 44 (70.5%) patients had lymph node involvement. The lymph nodes that had a high metastatic rate included the following: (1) lymph nodes around the common hepatic artery (number 8 lymph node); (2) lymph nodes of the hepatoduodenal ligament (number 12 lymph node); (3) the posterior pancreaticoduodenal lymph node (number 13 lymph node); (4) lymph nodes around the superior mesenteric artery (number 14 lymph node); (5) para-aortic lymph nodes (number 16 lymph node); and (6) the anterior pancreaticoduodenal lymph node (number 17 lymph node). Twenty-eight of these 31 patients had disease in the posterior pancreaticoduodenal lymph node. The patterns of lymph node involvement consisted of four combinations: number 13-number 17, number 13-number 14, number 14-number 16, and number 17-number 8. All of the patients with number 16 nodal involvement had number 14 lymph node metastasis. However, there was no relationship between tumor size and lymph node involvement. Based on these results, the main lymphatic pathway from the head of the pancreas to the para-aortic lymph nodes was thought to be via the lymph nodes around the superior mesenteric artery, assuming that lymphatic flow is anterograde. In addition, this study demonstrates that it is necessary to perform an extensive lymph node dissection, including the para-aortic lymph node, even in patients with small tumors.


Mucinous eccrine carcinoma of the eyelid. Mucinous eccrine carcinoma (MEC) is a rare malignant tumor that typically arises in the periorbital area. The authors report the 73rd case of primary MEC of the skin. This patient's clinicopathologic findings and the world literature are reviewed. Patient ages range from 8-84 years (median, 63 years). The male-to-female ratio was approximately 2:1. The racial distribution was 67% white, 32% black, and 4% Asian. Primary MEC originates in the head and neck region in approximately 75% of patients. The most common location was the periorbital area (40% or 29 of 73 patients). The local recurrence rate after conventional surgery was: eyelid, 34%; scalp, 36%; and face, 33%. Primary MEC is often a slow-growing tumor that may recur after traditional surgical excision. Recurrent eyelid MEC tends to be locally destructive with a regional metastatic rate of 3.5% (1 of 29). The regional metastatic rate for all sites was 11%, and the distant metastatic rate was 3%.


Immunocytochemical localization of estrogen and progesterone receptors in imprint preparations of breast carcinomas. The role of mammography as an effective means of early detection of breast cancer is well established. Needle localization of occult tumors under mammographic guidance has become the principle method of sampling impalpable breast lesions. As a result, the number of clinically inapparent breast carcinomas sampled has increased rapidly. Under these circumstances, as a result of the size and the nature of these "early carcinomas," it may not be possible to assess the hormone receptor expression of these tumors by conventional biochemical assay and/or immunocytochemical analysis. To determine the usefulness of imprint preparation for detecting hormone receptors, 214 examples of primary, recurrent, and metastatic breast cancers were studied. Immunostaining of imprint preparations with monoclonal antibodies against the estrogen (H222SPy) and progesterone receptors (KD68) showed a high degree of concordance with immunocytochemical assay and biochemical analysis done on frozen tissue of the same tumors. This technique can be done easily and is well suited for tumors that are too small and/or ill defined to permit separate sampling for hormone receptor analysis.


Epithelial--stromal interactions in tumors. A morphologic study of fibroepithelial tumors of the breast. The known spatial interaction between normal breast epithelium and its surrounding stroma prompted an investigation of the spatial relationship between stromal mitoses and the epithelial component of fibroepithelial tumors of the breast. The authors applied a novel computerized morphometric technique to routinely processed histologic sections of 23 fibroepithelial tumors (13 fibroadenomas and 10 phyllodes tumors). The proportional area of epithelium in successive concentric annuli surrounding stromal mitoses was measured, and its distribution was compared with that around suitable control points. The authors found that stromal mitotic activity in these tumors was significantly more likely to occur close to rather than remote from the epithelial component, with a significant excess of epithelium around mitoses compared with control points within a range of 79 microns. Essentially similar findings were obtained when randomly identified fibroblast nuclei were used as control points, thus obviating variations in stromal cell density with distance from epithelium as an explanation for the findings. These findings support the hypothesis that stromal growth in fibroepithelial tumors depends, to a variable extent, on the epithelial component. An interaction in the opposite direction (i.e., the stroma providing the growth support to the epithelium) also may occur, but this was not investigated. It is suggested that there is an interdependence of growth between the epithelial and stromal components in these tumors that explains their complex morphology and that stromal dependence on epithelium is lost with increasing malignancy of the stromal elements.


Human papillomavirus 16 exhibits a similar integration pattern in primary squamous cell carcinoma of the penis and in its metastasis. Human papillomaviruses (HPV) are among the most common causes of sexually transmitted viral infections in the United States, and HPV types 16, 18, and others have been strongly linked with the development of cervical cancer. DNA from these oncogenic HPV types also has been detected in biopsy specimens of penile intraepithelial and invasive neoplasms, indicating a causal role of these viruses in the malignant transformation of these tissue. Southern blot analysis and two-dimensional gel electrophoresis were used to investigate the presence and physical state of HPV in a patient with metastatic penile carcinoma. The presence of HPV 16 DNA integrated into the host's genome was documented in a primary penile squamous cell carcinoma and its lymph node metastasis. The identical restriction endonuclease cleavage patterns for HPV 16 in both the primary tumor and its lymph node metastasis indicate that both tumors arose from a single clonal event. This finding provides evidence of a causal role of HPV in squamous cell carcinoma of male genitalia.


Detection of human papillomavirus DNA in esophageal carcinoma in Japan by polymerase chain reaction. Human papillomaviruses (HPV) have been implicated strongly in the pathogenesis of human squamous cell carcinomas, especially of anogenital carcinomas. Some pathologic changes of the esophagus may be one of the candidates for HPV etiology, but the role of HPV infections in the carcinogenesis of the esophagus remains to be clarified. To elucidate the association of HPV with carcinogenesis of the esophagus, 45 biopsy samples of esophageal squamous cell carcinomas were examined for the presence of HPV DNA by polymerase chain reaction (PCR). Primers for PCR were (1) consensus primers (CP) for the simultaneous amplification of the E6-E7 regions of cancer-associated HPV types (HPV 16, 18, 31, 33, 52b, and 58), which have been shown to have transforming activities; (2) type-specific primers (SP16, SP18) for the E7 regions of HPV 16 and HPV 18, respectively; and (3) general primers (GP) for the simultaneous amplification of the L1 regions of HPV 6, 11, 16, 18, 31, and 33. PCR using CP first was done for screening and showed that 3 (6.7%) of 45 specimens contained HPV 16 or HPV 18 DNA, the oncogenic high-risk HPV types. This was confirmed by SP16 and SP18 PCR. However, no HPV DNA was detected by PCR using GP. These results suggested that the HPV DNA detected might be integrated into the cell genome with their transforming genes retained and their late regions deleted. Most oncogenic types of HPV (HPV 16 and HPV 18) were detected by PCR in carcinomas of the esophagus. Thus, HPV might play a role, although at a low frequency, in carcinogenesis of the esophagus.


Effects of preoperative chemotherapy on gastric adenocarcinomas. A morphologic study of 25 cases. Surgical neoadjuvant therapy for gastric adenocarcinoma affords the opportunity to evaluate critically the histologic effects of preoperative chemotherapy. Morphologic alterations in gastric adenocarcinomas were examined in the surgical-resection specimens from 25 patients after 6 weeks of preoperative chemotherapy. The group included 1 patient with a complete response; 4, with subtotal responses; 4, with partial responses; and 16, with no response to preoperative chemotherapy. Histologic manifestations of preoperative chemotherapy included mucosal edema, aggregates of histiocytes in the submucosa and muscularis externa, and stromal fibrosis of the submucosa and muscularis externa. Cytologic manifestations were uncommon and included a single case of signet ring cell carcinoma with diminution of the cytoplasmic vacuoles after preoperative chemotherapy. Clinical follow-up was limited, but 3 of the 25 patients died within 5-8 months after the diagnosis of gastric adenocarcinoma. The gastric-resection specimens from these three patients did not show any histologic manifestations of preoperative chemotherapy. As in tumors at other sites, the efficacy of surgical neoadjuvant therapy for gastric adenocarcinoma can be assessed, based on the histologic response of the resected tumor to preoperative chemotherapy.


Mantle zone lymphoma in a gastric glomus tumor. A 61-year-old man had an ileocolectomy for resection of an obstructing lesion of the terminal ileum, which proved to be a mantle zone variant of intermediate lymphocytic lymphoma. At laparotomy, an intramural nodule in the gastric antrum was observed; on resection, this was found to be a typical gastric glomus tumor, focally infiltrated by lymphoma. This combined tumor has not been described previously, to the knowledge of the authors, and could be misdiagnosed easily, although both components should be considered in the differential diagnosis of small cell gastric neoplasms and can be identified readily by immunohistochemical studies.


A phase I dose escalation trial of yttrium-90 microspheres in the treatment of primary hepatocellular carcinoma. Ten patients with primary hepatocellular carcinoma were treated with intraarterial instillation of yttrium-90 (Y-90) microspheres, including eight men and two women (median age, 52 years; range, 29-69 years). Four patients were treated at a targeted hepatic dose of 50 Gy, two at 75 Gy, and four at 100 Gy. In 8 of the 10 patients, there was a significant concentration of Y-90 in localized tumor masses with tumor-to-liver perfusion ratios from 1.0:1-10.0:1. No patient had a complete or partial response, but 10 patients had stable disease (median duration, 10 weeks; range, 5-64 weeks). The median survival was 18 weeks (range, 2-150 weeks), and three patients lived longer than 1 year. Significant bone marrow or hepatic toxicity was not seen. One patient had a radiation-induced duodenal ulcer that required surgical management. Intraarterial instillation of Y-90 microspheres appears to be safe and deserves additional evaluation to determine whether there is meaningful activity in patients with primary hepatocellular carcinoma.


Hepatitis C virus infection in hepatocellular carcinoma. Detection of plus-strand and minus-strand viral RNA. Although serum antibody to hepatitis C virus (anti-HCV) is found in many patients with hepatocellular carcinoma, the actual roles of HCV in carcinogenesis are unknown. With reverse transcription followed by the polymerase chain reaction (RT-PCR), HCV RNA was examined in the sera and liver tissues of 16 patients with hepatocellular carcinoma who did not have hepatitis B virus markers, 13 of whom had anti-HCV. In the 13 patients with anti-HCV, the HCV genomic plus-strand RNA was detected more frequently in noncancerous tissues (8 patients, 61.5%) and in sera (6 patients, 46.2%) than in cancerous tissues (3 patients, 23.1%). No viral RNA was found in either sera or tissues in the three patients without anti-HCV. By using a sense primer for the RT in the RT-PCR assay, amplification was attempted of a putative complementary minus-strand RNA that is believed to reflect viral replication in the eight patients with the plus-strand RNA. The minus-strand RNA was found in the noncancerous tissues of six patients; it was not detected in the cancerous tissues. These results suggest that HCV can infect and replicate predominantly in noncancerous cells but rarely in tumor cells.


Do the tumor cells of hepatocellular carcinomas dislodge into the portal venous stream during hepatic resection? The current study was undertaken to investigate whether or not tumor cells are dislodged into the portal venous stream during hepatic resection for hepatocellular carcinomas. A catheter was placed using echo guidance into the portal branch through the mesenteric vein in 31 patients. Cytologic examinations were done on multiple blood samples at various operative stages. Tumor cells were recovered in 7 of 31 patients in whom the tumor sizes were more than 5 cm and portal invasions were found microscopically and/or macroscopically. By contrast, the remaining 24 tumors were less than 5 cm in size and showed negative portal invasions. Recovery of the tumor cells was found, not during the earlier operative stage of mobilization or rotation of the hepatic lobe, but during the later stages of hilar dissection or hepatic parenchymal dissection. The portal pedicles should be divided before hepatic dissection in segmentectomy and lobectomy to lessen the chance of dissemination of intravasated tumor cells.


Neuropeptide Y secretion from a malignant extraadrenal retroperitoneal paraganglioma. A patient with a malignant extraadrenal retroperitoneal paraganglioma had elevated levels of immunoreactive neuropeptide Y (NPY) in the peripheral blood (5988 pg/ml; normal, 123 +/- 30 pg/ml [mean +/- standard error of the mean]). A 6-month course of chemotherapy allowed surgical removal of the previously unresectable primary tumor. Postoperatively, the plasma NPY level initially fell to 1089 pg/ml; continued chemotherapy caused an additional decrease to 440 pg/ml. Four months after surgery, the plasma NPY level increased to 940 mg/ml, coincident with hepatic metastases. This case is the first report of a NPY-secreting clinically nonfunctional malignant extraadrenal paraganglioma. Determination of circulating NPY levels may be useful in the diagnosis and follow-up of patients with neuroendocrine tumors.


Molecular characterization of congenital mesoblastic nephroma and its distinction from Wilms tumor. Congenital mesoblastic nephroma (CMN) is a rare tumor of the neonatal kidney. It was once thought to be a variant of Wilms tumor that also arises from primitive renal cells. Molecular characteristics of two CMN were studied to clarify their potential relationship to Wilms tumors. Patterns of gene expression were assayed by Northern blot hybridization analysis. Tumors were tested for loss of heterozygosity (LOH) at chromosomes 11p13 and 11p15 using Southern blot analysis. The CMN, like Wilms tumors, demonstrated high-level expression of insulin-like growth factor II. Unlike Wilms tumors, however, the CMN expressed neither the N-myc oncogene nor the putative Wilms tumor suppressor gene, WT1. Using a panel of probes spanning 11p13 and 11p15, no LOH was detected in the CMN, nor was there evidence of deletion or rearrangements of WT1. Although Wilms tumor and CMN both arise from the developing kidney, molecular characterization suggests that different factors are involved in the pathogenesis of these two tumors.


Aggressive inflammatory pseudotumor of the abdomen 9 years after therapy for Wilms tumor. A complication, coincidence, or association? Inflammatory pseudotumor (IPT) is a benign lesion that occurs in various organs and tissues. It is usually sharply demarcated from the surrounding tissue and surgery is considered to be the best treatment. This article discusses a 15-year-old boy with an aggressive IPT of the abdomen occurring 9 years after therapy for Wilms tumor. IPT widely involved the esophagus, stomach, and liver, producing severe dysphagia. Histologically, it showed classic features of IPT and, also, areas of metaplastic bone, a new feature recently described in the same lesion of the kidney. It mimicked malignant tumor clinically and led to extensive surgery, but early follow-up has shown no recurrence. Although many complications of surgery and chemotherapy are well known, the authors believe that it is unlikely to be the cause of IPT in the case presented. Therefore, the possibility of coincidence or association of Wilms tumor and IPT remains open.


Ultrastructural study of experimental myocarditis induced by cardiovirus (EMCV-M) in swine. Eight 6-week-old piglets were inoculated with a strain of encephalomyocarditis virus (EMCV) isolated from an outbreak which occurred naturally in the Po Valley in 1988. Two non-infected animals, kept in the same cage, were used as controls. Out of the eight inoculated piglets, two died and two were suppressed on the 2nd post infection day (PID), the four remaining were killed on the 5th, 7th, 11th and 15th PIDs. Control animals were killed at the end of the experiment. The pathogenesis of myocarditis has been studied using routine methods (Alcian-PAS, Masson's trichrome, Gomori's for reticulin and Mallory's stain), histochemical techniques (ATPase and NADH-TR reactions) and ultrastructural observations (TEM). All the inoculated piglets showed macro and/or microscopic lesions of lymphocytic myocarditis, only in one case associated with fibrinous exudation. One control piglet also showed myocarditic lesions, probably due to a contact infection. An early myocardial fibrosis was already present on the 5th PID. Ultrastructurally the cardiac muscle cells showed severe myofibrillar losses and other regressive alterations. Only on the 15th PID did we observe calcification of the degenerating myocytes, while ultrastructurally we detected needle-like calcium deposits in the mitochondria from the 5th PID. From the 5th PID in the areas of myocarditis the myocytes showed a reduction and/or absence of ATPase and NADH-TR reactions. On TEM, one or more aggregates of viral particles in crystalline array were detected in the cytoplasm of many endothelial cells.


Demonstration of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity in the rat cardiac muscle by cobalt-based cytochemistry. We have utilized cobalt-reaction technique for histochemical and cytochemical demonstration of ouabain-sensitive, K(+)-dependent p-NPPase (Na(+)-K(+)-ATPase). The incubation medium consisted of: cobalt chloride, tricine buffer, p-nitrophenylphosphate, KCl, and phenylalanine. Final pH 7.4. Ultracytochemically, reaction products were localized along the internal side of sarcolemma and its vesicles, T-tubule membrane, and capillary endothelial cells. These results suggest that the method is reliable and can be used to investigate the localization of Na(+)-K(+)-ATPase activity.


Micropharmacology of monoclonal antibodies in solid tumors: direct experimental evidence for a binding site barrier. Monoclonal antibodies (MAbs) often distribute nonuniformly in tumors. In part, that observation reflects intrinsic heterogeneity within the tumor; in part, it reflects poor penetration through tumor substance. Several years ago, we proposed the "binding site barrier" hypothesis (J.N. Weinstein, R.R. Eger, D.G. Covell, C.D.V. Black, J. Mulshine, J.A. Carrasquillo, S.M. Larson, and A.M. Keenan, Ann. NY Acad. Sci., 507: 199-210, 1987; K. Fujimori, D.C. Covell, J.E. Fletcher, and J.N. Weinstein, Cancer Res., 49: 5656-5663, 1989), the idea that antibodies (and other ligands) could be prevented from penetrating tumors by the very fact of their successful binding to target antigen. Calculations suggested that this might be a significant factor in the therapy of even microscopic nodules. The higher the affinity and the higher the antigen density, the greater the barrier. Here, we provide direct experimental evidence of such a barrier to the percolation of D3 MAb through intradermally implanted line 10 carcinoma of guinea pigs. After affinity purification using glutaraldehyde-fixed line 10 cells, the D3 had an average immunoreactivity of 88%, a binding constant of 1.6 +/- 0.3 (SEM) x 10(10) M-1, and saturation binding of 355,000 +/- 15,000 molecules/cell. Using a combination of double-label autoradiography and double-chromagen immunohistochemistry, we determined simultaneously the distribution of (a) i.v. injected D3 MAb; (b) coinjected isotype-matched control IgG (BL3); (c) D3 antigen; (d) blood vessels. The previously developed mathematical models aided in the design of these experiments. Double immunochemical staining of the tumors showed antigen-rich patches 100-800 microns across, surrounded by blood vessels. At a low MAb dose (30 micrograms), binding to antigen severely hindered penetration into antigenic patches as small as 200 microns, even at 72 h. Explanation of this finding by a physical barrier was ruled out by the observation that BL3 distributed uniformly in the same patches. At a higher dose (1000 micrograms), the binding site barrier could be partially overcome. The same general principles of micropharmacology may apply to biological ligands other than antibodies, including those secreted by genetically modified cells.


Development of liver tumors in transforming growth factor alpha transgenic mice. We studied the development of liver tumors in male transforming growth factor alpha (TGF-alpha) transgenic mice of the CD1 strain and examined the expression of the transgene by immunohistochemistry and in situ hybridization. Livers of 4-5-week-old transgenic mice contained areas of centribobular hypertrophy with low glucose-6-phosphatase activity. These areas progressively expanded, and hypertrophy and dysplasia became generalized in livers of mice at 10-12 months of age. The expression of the transgene, determined by either immunohistochemistry or in situ hybridization, was uneven in animals that were 10 weeks old or older. The positive hepatocytes formed patches with a predominant centrilobular distribution. We studied a total of 23 liver tumors (7 hepatocellular carcinomas and 16 adenomas) obtained from 11 mice at 13-15 months of age and from one 7-month-old animal which received zinc sulfate to induce the transgene. The carcinomas were well differentiated tumors, without glucose-6-phosphatase or gamma-glutamyltranspeptidase activity, that developed from the dysplastic parenchyma and occasionally within an adenoma. In all carcinomas and in 56% of the adenomas there was overexpression of the transgene in relationship to the surrounding tissue. The majority of the tumors that overexpressed TGF-alpha were alpha-fetoprotein positive, while alpha-fetoprotein staining was not detected in tumors (all adenomas) that did not show excessive transgene expression. We conclude that TGF-alpha functions as a promoter of liver carcinogenesis through its effect as an autocrine inducer of hepatocyte proliferation. Further, the data indicate that TGF-alpha overexpression may favor tumor progression.


Adoptive transfer of V beta 2-deleting activity with host cells from mice implanted with C4 preneoplastic hyperplastic alveolar nodules. Selective deletion of mature peripheral V beta 2+ T-cells was observed in BALB/c mice implanted with the syngeneic C4 preneoplastic hyperplastic alveolar nodule (HAN) but not in mice with sham surgery (W. Z. Wei, R. Ficsor-Jacobs, S. J. Tsai, and R. Pauley, Cancer Res., 51:3331-3333, 1991). We now report the participation of host cells in that process. Peripheral V beta 2+ T-cells were reduced by 50% or more 4 weeks after an i.v. injection of 10 x 10(6) spleen cells from C4 HAN-bearing mice. Both T- and B-cell-enriched splenocytes mediated V beta 2 deletion. Secondary adoptive transfer of splenocytes from the recipients of the primary adoptive transfer also resulted in V beta 2+ T-cell deletion. The splenocytes lose V beta 2-deleting activity after 500 rad irradiation. V beta 2 deletion induced by either C4 HAN or splenocytes was more profound in CD4+ than in CD8+ T-cells. Loss of V beta 2+CD4+ T-cells was observed 5 days after the adoptive transfer of splenocytes, whereas V beta 2+ CD8+ cells were not reduced until day 9 or later. The differential rate of V beta 2+ CD4+ and CD8+ T-cell reduction continued for at least 7 weeks after the adoptive transfer. The pattern of V beta 2 deletion and the sequence of T-cell loss is similar in the recipients of C4 HAN or of adoptively transferred splenocytes. Southern blot analysis demonstrates non-germ line Mtv or MMTV proviral DNA in C4 HAN. Splenocytes of C4 HAN-bearing mice express a higher level of 1.7-kilobase long terminal repeat transcript than normal BALB/c splenocytes, suggesting a role for a unique Mtv/MMTV provirus in V beta 2 deletion.


CD24, a signal-transducing molecule expressed on human B cells, is a major surface antigen on small cell lung carcinomas. Cell lines derived from human small cell carcinoma of the lung express high levels of a surface polypeptide termed the cluster-w4 antigen, which was previously identified as a potential target for toxin-based immunotherapy of lung cancer. We have cloned a complementary DNA encoding the cluster-w4 antigen from COS-1 fibroblasts transfected with a SW2 small cell carcinoma library, by panning with a mixture of the cluster-w4-specific monoclonal antibodies SWA11, SWA21, and SWA22. The sequence of the cluster-w4 complementary DNA encodes an unusually short (80-amino acid) protein identical to that recently reported for the leukocyte activation molecule CD24 except for a single valine-alanine substitution due to a single-base polymorphism within the region of the gene coding for the extracellular domain. Biochemical analyses of the cloned cluster-w4 antigen confirmed both the presence of the phosphatidylinositol tail and the extensive glycosylation reported for the CD24 molecule. Furthermore, the cloned cluster-w4 antigen expressed on COS cells was shown to react with a comprehensive panel of CD24-specific monoclonal antibodies, as assessed by indirect immunofluorescence staining. Northern blot hybridization indicated the presence of several transcript sizes for the cluster-w4 antigen that were greatly overexpressed in small cell carcinoma cell lines, compared with normal hemopoietic cells and CD24-positive cell lines. Southern blot hybridization of restriction digests of genomic DNA identified a complex pattern of bands consistent with either a complex gene structure containing many exons or the presence of a family of closely related genes.


Endogenous tumor necrosis factor exerts its protective function intracellularly against the cytotoxicity of exogenous tumor necrosis factor. Based on the findings that expression of endogenous tumor necrosis factor (enTNF), which is not present in TNF-susceptible cells, was generally observed in TNF-resistant cells and that TNF gene transfection gives rise to TNF resistance, the assumption was made that enTNF may be a protective protein against the cytotoxicity of exogenous TNF. However, it remains unknown whether the protection by enTNF is exerted in an intracellular or extracellular (autocrine) manner. We therefore transfected a nonsecretory human TNF gene (pTNF delta pro) into highly TNF-sensitive mouse tumorigenic fibroblasts (L-M cells) and investigated their TNF susceptibility. The transfectants expressed enTNF which was not secreted into the medium and acquired an appreciable degree of resistance to exogenous TNF. A significant increase in the manganous superoxide dismutase level was also noted in the transfectants. These findings suggest that enTNF exerts its protective function intracellularly by inducing manganous superoxide dismutase production.