Objective. To observe the regulating effects of vascular endothelial growth factor (VEGF) and angiotensinⅡ (ANG II) on the frog's pericardium, lymphatic stomata and angiogenesis so as to reveal their effects and mechanism on the mesothelial permeability, lymphatic stoma regulation and myocardial hypertrophy. Methods. VEGF and ANGⅡ were injected into the frog's peritoneal cavity so as to examine the changes of the pericardial stromata by using transmission electron microscopy, scanning electron microscopy and computerized imaging analysis. Results. Scattered distributed pericardial stomata were found on the parietal pericardium of the frog with a few sinusoid mesothelial cells, whose blood supply was directly from the cardiac chambers flowing into the trabecular spaces of the myocardium (because there are no blood vessels in the myocardium of the frog). The average diameters of the pericardial stomata in VEGF and ANGⅡ groups were 1.50 μ m and 1.79 μ m respectively, which were much larger than those in the control group (0.72 μ m, P< 0.01); the average distribution densities of the stomata were 8.25/0.1 mm2 and 12.80/0.1 mm2 in VEGF and ANGⅡ groups, which were also much higher than those in the control group (3.57/0.1 mm2, P< 0.01); the sinusoid areas in VEGF and ANGⅡ groups were 2442.95 μ m2/0.1 mm2 and 2121.79 μ m2/0.1 mm2, which were larger than that in the control group (995.08 μ m2 /0.1 mm2 , P< 0.01); no angiogenesis was found in the frogs of the experimental groups. Conclusions. VEGF and ANGⅡ could strongly regulate the pericardial stomata by increasing their numbers and openings with larger diameters and higher distribution density. They could also increase the sinusoid areas with the result of the higher permeability of the pericardium, which clearly indicated that VEGF and ANGⅡ could speed up the material transfer of the pericardial cavity and play an important role in preventing myocardial interstitial edema. Yet there was no strong evidence to show the angiogenesis in the myocardium.

Objective. The present study investigated the role of calcineurin in angiotensin II(AngII) induced cardiac myocyte hypertrophy of rats. Method. The primary cardiac myocytes were cultured under the standard conditions. The calcineurin activity in AngII treated cardiomyocytes was tested by using PNPP;protein synethsis rate was assessed by 3H leucine incorporation; atrial natriuretic factor(ANF) Mrna level was determined by Northern blot analysis. Cell viability was estimated by lactate dehydrogenase(LDH) levels in cultured medium and by dyed cell numbers. Result. After stimulation of 10,100 and 1 000nmol/L of AngII, calcineurin activities in the cardiomyocytes were increased by 13％ ,57％ (P< 0.05) and 228％ (P< 0.01) respectively, compared with control group. Cyclosporin A(CsA), a specific inhibitor of calcineurin, markedly inhibited the calcineurin activity and decreased the 3H leucine incorporation in AngII treated cardiomyocytes in a dose dependent manner. It was also found that CsA slightly reduced the Mrna level of ANF gene in AngII stimulated cardiomyocytes. Conclusion. During AngII induced cardiac myocyte hypertrophy, calcineurin signal pathway is activated, and inhibition of the pathway can attenuate AngII induced cardiac myocyte hypertrophy, which suggests that the calcineurin signal pathway may play an important role in AngII induced myocardial hypertrophy of rats.

Objective. To detect spirochetes in sections and to study the therapeutic effect of penicillin in cheilitis granulomatosa (CG) and Melkersson-Rosenthal syndrome (MRS) and the relationship between the spirochetes infection and CG and MRS. Methods. Routine HE sections and Warthin-Starry special staining were carried out in 20 cases of CG and 6 cases of MRS. Meanwhile there were 9 cases of CG and 2 cases of MRS were treated by penicillin (12 000 000u,IV, per day) for two courses (14 days). Results. A kind of spirochete was discovered in the sections of all cases of the CG and MRS .The CG and MRS could be divided into two types histopathologically, that is granuloma type and interstitial inflammatory type (non-granuloma type), those morphological changes tallied with spirochetosis. After treatment by penicillin, the facial and labial swelling of the 11 cases of CG and MRS were abated. Conclusion. CG and MRS probably are infectious diseases caused by spirochetes.

Objective. To evaluate the role of glucose transporter-l (GLUT1) in the glucose uptake of glomerular mesangial cells. Methods. Cultured C57/SJL mouse mesangial cells were used in the study. The expression of GLUT1 mRNA was detected by RT-PCR. The expression of GLUT1 protein was detected by immunofluorescence and flow cytometry. The uptake of glucose and its kinetics were determined by 2-deoxy-[3H] -D-glucose uptake. Results. Both GLUT1 mRNA and protein were found in mouse glomerular mesangial cells. 2-deoxy-D-glucose uptake and kinetics assay showed that this glucose transporter had high affinity for glucose and the glucose uptake specificity was further confirmed by phloretin. Conclusion. Functional GLUT1 did present in mouse mesangial cells cultured in vitro and it might be the predominant transporter mediated the uptake of glucose into mesangial cells.

Objective. To evaluate the effects of epidermal growth factor (EGF) on intestinal permeability and bacterial translocation in rats with acute pancreatitis during total parenteral nutrition (TPN). Methods. Thirty-two male Sprague-Dawley rats that underwent injection of 3.5％ sodium taurocholate solution into the pancreatic duct were randomly divided into one of the following two groups: (1) received only TPN (control group) or (2) received TPN with EGF at a dose of 0.2 mg· kg-1· day-1 (Egf group). On fifth day of total parenteral nutrition, samples from mesenteric lymph nodes, pancreas, liver and spleen were harvested for cultures. Water, protein and DNA content in jejunal mucosa were determined. D-xylose and fluorescein isothiocyanate (FITC)-dextran were instilled into the lumen of a ligated segament of small intestine. Thirty minutes later, superior mesenteric vein D-xylose and plasma FITC-dextran concentration were measured. Results. Positive cultures in liver and spleen, as well as FITC-dextran concentration in the Egf group were significantly lower than in the control group. Protein and DNA content in jejunal mucosa in the Egf group were significantly higher than in the control group. Conclusion. The results indicate that EGF may prevent increased intestinal permeability and bacterial translocation in rats with acute pancreatitis during TPN.

Objective. To study the redistribution of endothelin-1 (ET-1) receptors in two subcellular organelles , the sarcolemmal membrane and the light vesicle, of rat heart during the progression of sepsis. Methods. Sepsis was induced by cecal ligation and puncture (CLP). ET1 receptor was assayed by using [125I]-ET1 binding. Marker enzyme activities, protein yield, and dry-to-wet weight ratio of cardiac membranes were measured. Results. Septic rat heart exhibited two distinct phases: an initial hyperdynamic phase( 9h after CLP; early stage of sepsis) followed by a hypodynamic (18h after CLP, late stage of sepsis) phase. [125I]-ET1 binding study showed that during early stage of sepsis, the Bmax of ET1 receptors was increased by 30％ in sarcolemma but decreased by 19％ in light vesicles, while during late stage of sepsis, the Bmax was decreased by 24％ in sarcolemma but increased by 38％ in light vesicles.The total binding of sarcolemma and light vesicles was increased by 25％ during early stage of sepsis but decreased by 17％ during late stage of sepsis. Conclusions. These data indicated that ET1 receptors in the rat heart were externalized from light vesicles to sarcolemmal membranes during early hyperdynamic phase while internalized from surface membranes to intracellular compartment during late hypodynamic phase of sepsis.

Objective. To improve clinical knowledge of Peutz-Jeghers syndrome. Methods. Eight patients with Peutz-Jeghers syndrome from 1984 to 1998 in our hospital were retrospectively reviewed and analyzed in the present study. Result. The result of this analysis showed that there were 4 patients appeared with family histories of Peutz-Jeghers syndrome. All of the included patients admitted to the hospital with various complications, and eventually received surgical interventions for these complications, among which, 6 of them had intestinal obstructions mostly (5/6) due to small bowel intussusception, and 2 of them suffered with hemafecia. Post-operative recoveries were generally satisfactory with zero mortality. Conclusion. Peutz-Jeghers syndrome is an uncommon digestive dominant hereditary disease. The diagnosis of it with history, symptoms, signs, and proper examinations usually is not difficult. Surgical interventions are necessary once complications occur.

Objective. To evaluate the respective value of dual-phase helical CT arterial portography (CTAP) and conventional angiography in preoperative predicting resectability of pancreatic ductal adenocarcinoma. Subjects and methods. Tumor resectability was prospectively evaluated in 54 patients with pathologically proven pancreatic ductal adenocarcinoma who later underwent surgery. Both dual-phase helical CT scanning and selective angiography were obtained in each patient preoperatively. For optimal enhancement of pancreas and major peripancreatic vessels, two catheters connected to an automatic injector via a Y-shaped tube were placed after selective angiography,one in celiac trunk, the other in superior mesenteric artery. Then the patient underwent dual-phase helical CTAP of pancreas and liver. The criteria of irresectability for CTAP include: tumor diameter≥ 5 cm,extrapancreatic invasion, distant metastases and vascular involvement(occlusion, stenosis or semicircular encasement of superior mesenteric artery, hepatic artery, splenic artery, celiac axis; portal vein, superior mesenteric vein or splenic vein). The results of both modalities were correlated with findings from surgery or pathology. Results. Thirty-eight of 54 patients had nonresectable disease. In prediction the irresectability, sensitivity, specificity, positive predictive value, negative predictive value and overall accuracy were 94.7％ ,100％ ,100％ ,88.9％ ,96.3％ respectively for helical CTAP and 63.2％ ,93.8％ ,96.0％ ,51.7％ ,72.2％ respectively for selective angiography. In assessing vascular involvements, dual-phase helical CTAP was also superior to selective angiography. Conclusion. Dual-phase helical CTAP is superior to angiography in assessing resectability of pancreatic ductal adenocarcinoma. The combination of the two modalities may further improve overall accuracy of assessment.

Objective. To evaluate the Plasmodium falciparum CTL epitope vaccines in HLA class I allele specific human cell lines that have high frequency among Chinese population. Methods. Synthesized oligonucleotides encoding for P.f. CTL epitope genes, constructed eukaryotic expression plasmids, transfected the minigenes into HLA class I allele specific human cell lines and identified endogenous expressing of the minigenes by RT-PCR and HLA stabilization assay. Results. Two mini-genes encoding Plasmodium falciparum CTL epitopes were designed and cloned, respectively, into an eukaryotic expressing vector to form TR26 which was restricted to HLA-B51, SH6 which was restricted to HLA-A2.1, and TS, which had the two aforementioned mini-genes fused in tandem. All of these CTL epitope genes were transfected and endogenously expressed in respective cell lines containing appropriate HLA molecules. The obviously increased expressions of HLA class I molecules were detected in the transfected cell lines. It was demonstrated that the two discrete Plasmodium falciparum epitope genes were effectively processed and presented, and the close proximity of the two epitope genes in one chain as in mini-gene TS did not interfere with the processing and presenting of each epitope gene in corresponding cell line. Conclusion. A successful expression and presentation of multiple CTL epitope mini-gene in MHC class I allele specific human cell lines were demonstrated by an in vitro assay, which could be corresponding to the vaccination of CTL vaccines in people with different MHC I molecules. This work also suggested the possibility of constructing a multiple CTL epitope plasmodium falciparum DNA vaccine that could cover most of Chinese population.

Objective. To investigate the morphological properties of pig cerebral bridging vein. Methods. The morphology and fibre arrangement of 15 cerebral bridging veins obtained from 7 Danish Yorkshire landrace pigs were observed. Results. There was a narrow region at the junction of the cerebral bridging veins and superior sagittal sinus termed “ outflow cuff segment". The diameter and length of outflow cuff segment were much smaller and the thickness was higher than those of the cerebral bridging veins (P< 0.01), and circumferential collagen fibres were most dense in the outflow cuff segment. The opening angle of the outflow cuff segment and the cerebral bridging veins were 115± 4° and 120± 4° (P >0.05). Conclusions. There were differences in fibre arrangement and morphological properties between the outflow cuff segment and the cerebral bridging vein, just like a resistance valve, the outflow cuff segment may play an important role in stabilizing cerebral venous outflow and regulating intracranial pressure.

Objective. To investigate the association of carotid arterial intima media thickness (IMT) with principal cardiovascular risk factors in the elderly. Methods. Carotid arterial IMT was measured by high resolution B mode ultrasound in 94 elderly subjects (old aged group), and compared with subjects aged < 60 (middle-aged group). Results. In comparison with the middle-aged group, the prevalence of coronary heart disease, cerebral vascular disease, hypertension and diabetes mellitus was significantly higher, and serum cholesterol and systolic blood pressure were also significantly higher in old aged group. Although there was no obvious difference in IMT between the two groups, carotid plaque and carotid wall thickening were more frequently found in old aged group. Age, systolic blood pressure and serum cholesterol were shown as the independent determinants for carotid IMT in the total participants, whereas no such independent relation was found in old-aged group. Conclusion. Age is the major risk factor for carotid atherosclerosis in the elderly. In other words, the occurrence of carotid atherosclerosis is the result of advancing age combined with the effect of multiple cardiovascular risk factors.

Objective. To assess the effectiveness of the flashlamp-pumped pulsed dye laser (Photogeneca V, Synosure Corp, Boston, United States) in the treatment of port-wine stains. Methods. One hundred and ninety-four consecutive patients with port-wine stains were treated with a flashlamp-pumped pulsed dye laser in Peking Union Medical College Hospital from January 1998 to August 1999. Results. Of 194 patients who completed treatment, 56.2％ had more than 60％ fading of the lesion and only 6.7％ had less than 20％ fading. An average of 3.6 treatments were needed to achieve more than 60％ fading. The response was better in children than in adults, although the difference was not significant. Pigmentary change (usually transient) occurred in 3.1％ of patients. Conclusions. This study confirms the efficacy of the flashlamp-pumped pulsed dye laser in the treatment of port-wine stains in children and adults.

Objective. To determine the effects of HOE 694, a new and potent Na＋ -H＋ exchanger blocker, on transient inward current (Iti) and Na＋ -Ca2＋ exchange during hypoxia-reoxygenation in guinea pig cardiomyocytes. Methods. Cardiomyocytes were isolated from adult guinea pig ventricle. Experiment was performed in an experimental chamber that allowed the cells to be exposed to a sufficiently low O2 pressure. The cells were subjected to hypoxia and reoxygenation. The ionic currents were studied with patch clamp technique. Results. In the absence of HOE 694, hypoxia-reoxygenation induced Iti in 12 of 15 experiments; but in cardiomyocytes pretreated with HOE 694 (10～ 50 μ mol/L), the incidence of Iti observed during reoxygenation was reduced to 5 of 11 experiments and 3 of 10 experiments, P0.05 vs control respectively. The Na＋-Ca2＋ exchange current was unaffected by HOE 694 under normoxic condition. However, when cells were pretreated with 10 μ mol/L HOE 694 for 10 min, then subjected to hypoxia condition, the Na＋  Ca2＋ exchange current was significantly inhibited. Conclusions. Blockade of the Na＋ -H＋ exchange by HOE 694 could reduce Ca2＋ overload upon hypoxia-reoxygenation, and inhibition of Na＋ -H＋ exchange may also indirectly decrease Na＋ -Ca2＋ exchange activity during hypoxia.

T cell receptors (TCR) are a collection of several proteins located in the outer membrane of the T cell. Mature T cell has definitive TCR genes,which may be used as a clonal marker. To investigate whether TCR gene analysis may be useful for diagnosis of myasthenia gravis (MG) with thymoma, we used the cDNA of the β－ subunit of the TCR served as probes to analyze the structure of the TCR β gene and to determine the extent of diversity between MG with thymoma and MG without thymoma. Eighteen patients were clinically diagnosed generalized MG, eight of them had thymoma. Peripheral blood lymphocytes (PBL) were obtained at or shortly before thymectomy (except one, 2 years after thymectomy). When DNA was digested with EcoRI and hybridized with the 32P labeled probes, two germline bands of 10.5kb and 4.0 kb appeared in DNAs from all 18 patients. The 10.5 kb band contains the C β 1 region and the 4.0kb fragment the C β 2 region, respectively. The 4.0 kb band is usually reduced as compared with the 10.5kb band. This can be explained by the preferential rearrangement of the C β 2 gene in T cell with MG. We were able to demonstrate a rearranged band of 9kb in EcoRI－ digested DNAs of four patients. All the four patients had been found to have thymoma , of which 2 were lymphocyte and 2 mixed type. But rearranged bands could not be detected in the DNAs of another four patients with thymoma, of which 2 were epithelial and 2 mixed type. When DNA was digested with HindIII and BglII, germline bands appeared in DNAs from all patients. No rearranged bands could be observed . Single rearranged bands were detected in four patients with thymoma, this showed that specific TCR gene rearrangement may be related to certain types of thymoma . Detection of TCR gene rearrangement might be helpful to the early diagnosis of MG with thymoma . But we were unable to find gene rearrangement in another four thymoma (2 were epithelial and 2 mixed type). There are three possible explanations for the results. First, thymoma has heterogeneity. Second, TCR gene rearrangement only reflects the clonal origin of the T cell tumor. Third, in one patient, peripheral blood lymphocytes were obtained two years after thymectomy, neoplastic T cells maybe disappear. Thus, our results suggested that TCR gene rearrangement in patients with MG may be helpful for diagnosing certain type of thymoma.

DNA chip technology employs light-directed in situ oligonucleotide synthesis and/or DNA microarray printing device to produce arrays of large number of probes in the tiny surface of silicon substrates, which makes it possible that the gene detection be conducted efficiently with high speed and sensitivity. The DNA chip may take important part in genome research, gene diagnoses and so on.