Objective To study an effective method for culturing of human vascular smooth muscle cells (hVSMCs) in tissue engineered blood vessels (TEBVs) in vitro.Methods A 3-cm segment of human long saphenous vein was harvested under sterile conditions.The primary culture and subculture were done by modified tissue-piece inoculation and trypsin digestion respectively.The cells were purified by mechanical treatment and differential attachment.PDGF was combined with high concentration of glucose DMEM as the culture medium.The cultured cells were identified by morphology and immunohistochemistry with contrast microscope and SABC kit,and RT-PCR method detected the expression of ?-SMA and calponin 1.Results The cultured cells possessed"peak and valley"characteristics for VSMCs under microscope.Immunohistochemical staining showed positive expression of intracytoplasmic ?-actin,and RT-PCR detected positve expression of ?-SMA and calponin 1.Conclusions Culture medium of PDGF combined with high concentration of glucose DMEM,and with differential attachment method can provide highly purified hVSMCs with good structure and function.

[Objective] To study the correlation between kidney cell DNA degradation and postmortem interval within the span of 6-48 hour after the subject rats′ death.[Methods] To select 18 healthy mature female SD rats and equally divide them into 6 groups.To execute the rats with cervical spine articulation and put the rats under the incubator temperature of 25.1℃ (the average temperature of the 5 previous Decembers in Guangzhou prefecture).Sample kidney tissue from the rat separately 0 hour,6 hours,12 hours,24 hours,36 hours,48 hours,and 60 hours after the rats′ execution to prepare monoplast suspension,which is committed to comet assay.The comet images were captured by fluorescence CCD.Kinetic Comet 4.0 software was used to analyze images.Relevant data were collected by kinetic Comet 4.0 software and were subjected to Kruskal-Wallis test.[Results] Within the postmortem interval of 6-48 h,the number of SD rat kidney cell DNA fragments increased as the postmortem interval lengthens.So did the comet tail length.The Oliver tail moment and tail DNA of comet also showed sign of increase in positive proportion to the postmortem interval (their values corresponding to 60-hour-postmortem-interval were not obtainable.Kruskal-Wallis test indicated:the discrepancies of TL among the 6 groups were all significant (P ＜ 0.01).The difference of TM between 6 h group and 12 h group was not significant (P ＞ 0.05).The difference of TM between 24 h and 36 h was significant (P ＜ 0.05).The difference of TDNA among 6 h,12 h,and 36 h groups were not significant (P ＞ 0.05).The difference of TDNA between 36 h and 48 h was significant (P ＜ 0.05).[Conclusion] Degradation of nuclear DNA of the rat kidney cells increases as the postmortem interval lengthens and comet assay may provide important empirical evidence for determining the postmortem interval.

Objective To explore the reasonable construction of the bilingual teaching training mode for medical professional teachers so as to improve the effectiveness of the bilingual teacher training,promote the development of faculties and the quality of bilingual teaching in colleges and universities.Methods First,need analysis was done on 45 medical teachers who had taught medical scienc.e in English or intended to do it in Third Military Medical University and four basic modules were formed lectures in class,one on one instruction,self training,simulation teaching.Lectures in class are related to how to improve students' ability of language,make PPT and design teaching plans.One on one instruction focuses on pronunciation,lecture notes writing in English,trial teaching and teaching attitude;self training includes human-computer interaction in listening and speaking,independent preparation for lessons and video learning;simulation teaching is composed of trial lecture in class,teacher-student comments and demo lessons.Then 3 students participating the training was investigated and the analytic hierarchy process (AHP) was applied to calculate the weights of every indices.Results Primary index weights are:0.490 for one-on-one tutoring;0.240 for simulated teaching;0.180 for having English lessons;0.082 for independent learning.Weights of secondary indices are as the following.Trial teaching,the lecture notes writing in English and the training for English pronunciation and teachers' classroom language which are subordinate to the primary index of one-on-one tutoring are 0.180 0,0.150 0 and 0.096 0 respectively;teachers' comments and peer feed-back and demo class under the primary indices of simulated teaching are 0.120 0 and 0.091 0.Conclusion In order to improve the teaching ability in English,it is necessary to strengthen the one-onone tutoring for medical professional teachers,highlight English teachers' comments and peer assessment and excavate the effect of demo classes in the training program.

Objective Cathepsin K (CTSK) played an important role in adipocyte differentiation.The activation of CTSK needs to convey by mannose-6-phosphate receptors (M6PR) in osteoclasts. The aim of the present study was to identify the effects of mannose-6-phosphate (M6P) in adipocyte differentiation and its underlying molecular mechanism. Methods Oil red O staining, accumulation of cytoplasmic triglycerides and glycerine release were used to assess its effects on adipocyte differentiation in the 3T3-L1cell line. The enzyme activity of CTSK was observed by laser confocal microscopy. The proliferation of 3T3-L1 preadipocytes was detected by MTT methods. mRNA expression of M6PR was determined by RTPCR. Results M6P could prevent adipocyte differentiation in a dose-dependent manner as evidenced by absence of triglyceride accumulation and glycerol content. Statistical significance was showed when the concentrations of M6P were 5.0 mmol/L and 8. 0 mmol/L respectively(P ＜0. 05). The mRNA expression of M6PR was detected during the whole process of adipocyte differentiation. With the increase of M6Pconcentration, enzyme activity of CTSK was inhibited in a concentration-dependent manner. MTT method showed that the absorbance at 570 nm of 3T3-L1 preadipocytes was 0. 057 ±0. 091, increased about 62. 9%at 10. 0 mmoL/L compared with the control group (P ＜ 0. 05 ). Conclusion M6P inhibits the terminal differentiation of adipocyte, which may be associated with its effect of blocking CTSK activity by competitive binding with M6PR.

BACKGROUND: Neuroscience and brain science researches have paid attention to the effect of astragalus membranaceus in the treatment of neurologic impairment disease and neural regeneration. Studying astragalus membranaceus effects on neural stem cells are becoming a new research direction. OBJECTIVE: To explore the effects of astragalus injection on biological viability of rat neural stem cells. METHODS: Neural stem cells of Wistar rats were separated and cultured. Immunofluorescence staining was applied to identify the neural stem cells. The purified cells were gained by the second subcultivation in vitro, and then the cells were randomly divided into control group and astragalus injection groups with various concentrations (50, 200, 400 g/L) to culture for 6, 12 and 24 hours. The activity of cells was tested by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, and then the immunohistochemistry was applied to detect the expressions of neuron-specific enolase and glial fibril ary acidic protein in the 50 g/L astragalus injection group after induced for 7 days. RESULTS AND CONCLUSION: The viability of neural stem cells increased significantly after intervention with different concentrations of astragalus injection for 6 hours as compared with the control group (P < 0.05). However, there was no difference in the cel viability after treated with different concentrations of astragalus injection for 24 hours (P > 0.05). Compared with the control group, the cells in the 50 g/L astragalus group differentiated rapidly, and the number of positive cells for neuron-specific enolase was increased significantly (P < 0.05). The neural stem cells proliferation was hastened, and its differentiation was promoted by the interference of astragalus injection.

Objective To screen and identify the colonic adaptation-related proteins.Methods Totally 20 SD rats were equally randomized into 2 groups:ultra-short bowel group (90％ of intestine was resected) and control group (transection and then anastomosis of the intestine).After 21 days of feeding,two-dimensional electrophoresis (2-DE) was performed to separate the total protein from the colon mucous epithelial tissues.Then Image Master 2D Platinum software was use to analyze differential expressional proteins,which were further analyzed by MALDI-TOF-MS and Mascot software to identify these proteins.Results The 2-DE patterns with high resolution were obtained from the colon mucous epithelial tissues of both groups.Four differentially expressed proteins were identified by MALDI-TOF-MS:M2-PK,AGR2,proteasome activator complex subunit 1,and pancreatic triacylglycerol lipase.These proteins were involved in sugar and fat metabolism and cell proliferation.Conclusion The identified proteins may play important roles in the process of colonic adaptation through substance metabolism and by stimulating cell proliferation.

Objective To explore the characteristic manifestations of breast malignant isodense masses using mammography. Methods 121 breast isodense masses with pathological confirmations were collected.Compared indicators with pathological findings were the tumor size,shape,edge,structure changes around the mass,axillary lymph nodes,skin changes,nipple changes,suspi-cious malignant calcification.χ2 test and Logistic regression analysis were used to evaluate the diagnostic values of each indicator.Re-sults In the 121 isodense masses,73 were benign,48 were malignant tumors.According to χ2 test analysis,factors impacted the nature of mass including:tumor morphology (χ2 =14.376,P =0.002),tumor edge (χ2 = 21.555,P =0.000),structure twisted surrounding the mass (χ2 =26.939,P =0.000),axillary lymph nodes (χ2 =1 6.285,P =0.000),skin thickening (χ2 =4.698,P =0.030).According to multivariate binary Logistic regression analysis,risk factors of the Logistic regression model included:edge infiltrating of mass (P =0.014),structure twisted surrounding the mass (P =0.003),axillary lymph nodes (P =0.026).Conclusion The characteristic manifestations of breast malignant isodense masses include edge infiltrating,structure twisted surrounding the mass,axillary lymph nodes.These manifestations are of great significance in differential diagnosis of breast isodense masses.

OBJECTIVE A surveillance study was performed for nosocomial infections in order to investigate the change in antimicrobial resistance of Klebsiella pneumoniae,especially the strains isolated from 1999 to 2004.METHODS K-B test was used for the antibiotics susceptibility test and the results were read based on National Committee for Clinical Laboratory Standards(NCCLS) of the USA.The situation of ESBLs-producing strains of K.pneumoniae was investigated.RESULTS Totally 326 K.pneumoniae strains showed the highest susceptibility to imipenem.Ceftazidime,cefepime,and cefoperazone/sulbactam also showed excellent activity against K.pneumoniae.The prevalence of ESBLs from 326 strains was 20.2%.CONCLUSIONS It is important to study the drug resistance in nosocomial infections by K.pneumoniae.

This research adopted silt as the sample,and the five highest hydrogen production performing strains contained in the sample were isolated. The strain whose hydrogen production was the highest was identified as Enterobacter cloacae by the analysis of 16S rDNA sequencing and comparison. It is showed by Plackett-Burman Experimental Design that only glucose,citric buffer and reducing agent had significant effects on hydrogen production by Enterobacter cloacae FML-C1. The path of steepest ascent was undertaken to approach the optimal response region of those three factors. Central Composite Design(CCD) and Response Surface Methodology(RSM) were employed to investigate the interaction of the variables and to ascertain the optimal values of the factors,which finally led to the maximum hydrogen production(VH2) . The theoretical optimal medium conditions were:glucose 21.5 g/L,citric buffer 13.6 mL/L,reducing agent10.0 mL/L. The five tentative tests matched this model well. The final VH2 was up to 2347.4 mL/L,which was 127.42% enhanced in comparison to the original. The result shows that PB experiment design and RSM analytical method work well in selecting factors which have significant influences on the hydrogen production and,moreover,achieve the ideal optimal result.

Objective To investigate the glomerular microvascular injury and repair in patients with IgA nephropathy (IgAN) as well as its relationship with intermedin (IMD).Methods Eighty cases of renal tissue taken from patients first diagnosed as IgAN in Shanxi Provincial People's Hospital Affiliated to Shanxi Medical University and 15 cases of normal renal tissue were detected by the expression of glomerular IMD,CD31,and VE-cadherin through immunohistochemical method.ELISA method was used to detect VEGF and IMD of plasm from 31 normal subjects and 36 cases chosen from the IgAN patients.Their changes and internal relationship were analyzed according to Lee's and chronic kidney disease (CKD) classification.Results (1) Compared with the control group the expressions of CD31,IMD,and VE-cadherin in IgAN patients were statistically significant (P ＜0.01).Compared with the control group the levels of IMD and VEGF in plasma of IgAN patients in early stage of CKD group and late stage of CKD group were statistically significant (P ＜ 0.01).(2)Correlation analysis:the expression of glomerular CD31 and Lee's classification were negatively correlated (r=-0.232,P ＜ 0.05);glomerular IMD was negatively correlated with Lee's classification (r=-0.241,P＜0.05),while positively correlated with glomerular VE-cadherin (r=0.417,P＜ 0.01).VEGF in plasma of IgAN patients was positive correlated with CKD classification,BUN (r=0.458,0.409,P＜0.05),and negatively correlated with serum ALB (r=-0.532,P＜0.01).Conclusion Microvascular injury exists in patients with IgAN.The expression of VE-cadherin and IMD are positively correlated,suggesting that IMD may be involved in the progression of vascular protection and angiogenesis in IgAN.The contents of IMD and VEGF in plasma of IgAN patients increase,indicating that they may play a role in the progression of IgAN.