Objective The experiment aims at probing the best condition of the isolation and purification of rat islets.Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability.Results The histological staining revealed that the viability and the purity of the purified islets were above 95％and 85％respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There aye many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.

Objective The experiment aims at probing the best condition of the isolation and purification of rat islets. Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability. Results The histological staining revealed that the viability and the purity of the purified islets were above 95% and 85% respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There are many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.

Objective To observe the dynamic changes of urotensionⅡ (U Ⅱ) content in plasma and bronchoalveolar lavage fluid (BALF) of rats with chronic hypoxic pulmonary hypertension and to investigate the effects of hypoxia on the synthesis and secretion of U Ⅱ and the correlation of U Ⅱ with the mean pulmonary artery pressure (mPAP) and the partial pressure of arterial oxygen (PaO 2). Methods Male Wistar rats were randomly divided into control group and hypoxic groups. Model of chronic hypoxic pulmonary hypertension was established by normal barometric and discontinuous hypoxia. mPAP and RVdp/dt max of each rat were measured using right cardiac catheterization. PaO 2 was detected by blood gas analysis. U Ⅱcontents in plasma and BALF were detected by radioimmunoassay. Results During hypoxia, mPAP, RVdp/dt max , U Ⅱ content in plasma, and BALF increased in rats (P

Objective To understand the mechanism of how cardiomyocytes exit from the cell cycle,we examined the expression of polo-like kinase 1(plk1) in the postnatal developmental process of cardiac myocytes. Methods Mitotic Index(MI) of cardiomyocytes was examined in the neonatal,2-week-old,4-week-old,and adult rat hearts(five cases per groups) by double immunofluorescence stained with H3P and ?-sarcomeric actin antibodies.plk1 mRNA and protein expression during the postnatal developmental process of cardiac myocytes were detected by RT-PCR and Western blot analysis in rat hearts. Results The MI of cardiomyocytes in 0-day-old hearts(0.905?0.087%) was approximately 2.4 times over that in 2-week-old hearts(0.372?0.094%)(P

Objective To investigate the activity changes of gelatinase in the formation of ascending aortic aneurysm.Methods Thirty five young Wistar rats were divided into two groups:the control group and the experiment group.The rat models induced by ascending aorta banding were made.The ascending aortas were taken after 3-5 months operation,changes of gelatinase activity was observed by gelatin zymography and film in situ zymography.Results Gelatinase activity of ascending aortic aneurysm was significantly increased compared with that of normal ascending aortic aorta.Conclusion Elevation of gelatinase activity may play a significant role in the formation of ascending aortic aneurysm.

Objective To investigate the expressions and significance of transforming growth factor-beta 1(TGF?1) and its typeⅡ receptor(TGF?RⅡ) in experimental rat ascending aortic aneurysm of rat model.Methods The rat ascending aortic aneurysm models were made by banding ascending aorta of young Wistar rats.The ascending aortas were taken 4 months after operation.Immunohistochemistry staining and Western blotting were used to investigate the expressions of TGF?1 and TGF?RⅡ.Result Immunohistochemistry staining results showed that TGF?1 expressed in all layers of the aortic aneurysm and the control.TGF?RⅡ was extensively located in the hyperplastic intima and tunica media smooth muscle cells in the aortic aneurysm,while there was only a little positive staining in the control group.Western blotting results indicated that the expression levels of TGF?1 and TGF?RⅡ in the aortic aneurysm were stronger than the control,P

Objective To analyze the distribution and pulsed-field gel electrophoresis (PFGE)of pathogenic bacteria causing infectious diarrhea in a district of Beijing from 2011 to 2013,and provide basis for tracing infection sources.Methods A total of 1 179 stool specimens of infectious diarrhea from patients in a diarrhea outpatient department from January 2011 to December 2013 were collected,all isolated pathogens were identified by serotyping and PFGE analysis.Results 330 enteric pathogens were isolated from 1 179 specimens,the top 4 bacteria were Shi-gella spp .(28.18%,n=93),Salmonella spp .(20.91 %,n=69),Vibrio parahaemolyticus (13.33%,n =44),and diarrheagenic Escherichia coli (3.33%,n = 11 ).18 Shigella sonnei isolates were identified as 8 PFGE patterns, clustering similarity was close to 88%;69 Salmonella spp .strains belonged to 18 serotypes and 41 PFGE patterns, Salmonella senftenberg and Salmonella enteritidis had dominant patterns;no dominant PFGE patterns were obviously identified among 23 strains ofVibrio parahaemolyticus .Conclusion The serotypes and PFGE patterns of pathogenic bacteria in infectious diarrhea in past three years showed a wide distribution characteristics,the dominant PFGE patterns of Salmonella spp .and Shigella spp .need to be paid more attention,and outbreak of infectious diarrhea caused by Salmonella spp .and Shigella spp .should be alerted.

Objective To evaluate the growing condition of human umbilical vein endothelial cells (HUVECs), which were cultured on the membrane of different component, such as Chinese medicine, before or after the alkali treatment of 3-hydroxybutyrate-co-3-hydroxyhexanoate copolyesters (PHBHHx) and the biocompatibility between PHBHHx flim and endothelial cell. Methods The HUVECs were harvested by Baudine method,and identified by immunohistochemical method.Then the HUVECs of third passage were inoculated on the material surface and cultured for 8 hours,12 hours and 24 hours. After that, the morphology of HUVECs on different surfaces were observed by scanning electron microscopy, the distribution condition on different membranes were compared by cell-labeling immunofluorescence, and the cell viabilities of all groups were detected by MTT method. Results The HUVECs were successfully separated,and immunohistochemistry staining of FLK-1 and factor Ⅷ was positive.The result of HUVECs culture showed that cells on the material surface growed well, and proliferated significantly. The MTT analysis showed that the PHBHHx film of surface modification and adding some certain proportion of Chinese medicine could promote the growth and proliferation of HUVECs in vitro,and the cells were thriving, full shape, distribution on the surface by scanning electron microscopy and fluorescence microscopy.Conclusion The PHBHHx film of surface modification and containing certain proportion of Chinese medicine coating had good compatibility of HUVECs, which was favourable to cell growth, adherence and proliferation in vitro.

Objective To assess the interference by calcium dobesilatein 7 peroxidase-baseduric acid assays and to determine its clinical significance.Methods In the in vitro experiments, uric acid in pooled serum with final concentrations of calcium dobesilate additions (0, 2, 4, 8, 16, 32, and 64μg/ml) were measured by 7 peroxidase-based assays.Percent Bias (%) was calculated relative to the drug-free specimen.In the in vivo experiments, changes in serum uric acid and calcium dobesilate concentrations were observed before and after calcium dobesilate administration ( baseline, 0 h, 1 h, 2 h, 3 h, 4 h, 6 h ) involunteers.The interference in different assays was assessed compared with LC-IDMS/MS method. Calcium dobesilate levels in 40 specimens from those taking calcium dobesilate were measured by HPLC method.Of the 40 specimens, 10 were selected to analyse the levels of uric acid by both peroxidase and UV measurement method to assess the impact in clinical status.Results In the in vitro study, concentrations of uric acid measured by 7 peroxidase-based assays were reduced by -6.3%to -21.2%compared with drug-free serum, when theconcentration of calcium dobesilate was16μg/ml.In the in vivo study, comparedto UA levels at 0 h, the biasesof serum uric acid determined by peroxidase method after calcium dobesilate administration(1 h, 2 h, 3 h, 4 h, 6 h) were of -3.33%, -6.79%, -7.49%, -6.07%, -4.09%, respectively.The observed uric acid concentrations for 8 participants measured by enzymatic assays were inhibited by -3.75% to -6.89% at 0 hour and by -16.9% to-22.22% at 2 hours relative to the concentrations measured by the LC-IDMS/MS method. Conclusions Calcium dobesilate produced a clinically significant negative interference with uric acid in all peroxidase-based uric acid assays,which may result in false evaluation of uric acid level in clinical status.Significant differences in the degree of interference were observed among the assays.

Currently, there are two kinds of academic degree and professional degree in viral hepatitis postgraduate degrees. This kind of trainingmethod cannot combine clinical thinking and scientific research thinking. The combination of good research thinking and clinical foundation is beneficial to talent cultivation. In the future, the training of viral hepatitis graduate students requires students to have a clinical basis, and also requires students to have rigorous research thinking.