Objective:To obtain DNA of human Helicobacter pylori heat shock protein A, and construct a recombinant vector containing gene encoding HspA for nucleotide sequence analysis.Methods:The target gene was amplified from Hp chromosome by PCR, and then digested by restricted endonuclease enzyme of kpn I , BamH I simultanously, and inserted into the prokaryotic expression vector pET32a(+) digested by corresponding restricted endonuclease enzyme. The recombinant vector was selected and transformed for nucleotide sequence analysis.Results:Enzyme digestion analysis and sequencing showed that the target gene had been inserted into recombinant vector, but as compared with gene reported by GenBank. 1.4 % of the gene mutation and 1.6% of amino acid residues change in Hp happened respectively. The DNA sequence analysis showed the sequence of HspA DNA was almost the same as that published by GenBank.Conclusion:The gene coding for Hp HspA is cloned successfully.The results obtained lay the foundation for research on development of Hp protein vaccine and a quickly diagnostic kit applying to detection of Hp infection.

Objective:To construct a recombinant vector containing gene encoding heat shock protein A (HspA) and VacA with Mr 13 000 and 26 000 simultaneously from human Helicobacter pylori(Hp)for study on expression in E.coli BL21,and analysis of their antigenicity.Methods:The target genes encoding HspA and VacaA were amplified from Hp chromosome by PCR,digested by restricted endonuclease enzyme respectively,and inserted into the corresponding enzyme digested prokaryotic expression vector pET32a(+).The recombinant vectors pET32a(+)/HspA and pET32a(+)/VacA were used to select and transform for sequence analysis.After recombinant vectors digested by restricted enzyme of Xhol,BamH Ⅰ simultaneously,the pET32a(+)/HspA and VacA were taken out of agarose electrophoresis,and connected by T4 ligase again.The recombinant vector pET32a(+)/HspA-VacA was used to select and identified by PCR and restricted endonuclease enzyme.Results:Enzyme digestion analysis and sequencing showed that the target genes were found in 1080 base pairs,and had been inserted into recombinant vector,but as compared with gene reported by GenBank,3.40% of the gene mutation and 1.11% of amino acid residues change in Hp happened respectively.Conclusion:The genes coding HspA and VacA with Mr 13 000 and 26 000 respectively are cloned successfully.The results obtained lay the foundation for research on development of Hp protein and DNA vaccine applying to prevention of Hp infection.

Achievements for schistosomiasis control have been gained by implementation of integrated control strategies according to local conditions since the founding of the People′s Republic of China.By the end of 2008,5 of the 12 provinces reached the criteria of transmission interruption.Among 454 endemic counties,transmission was interrupted in 265 counties while 97 counties reached the criteria of transmission control.The number of schistosomiasis cases decreased from 10 million in history to 413 000.Currently,there still left 92 counties where the disease is endemic and mainly distributed in lake and mountainous regions.Limited by the environmental and socio-economic factors,integrated control strategies could not be carried out in these places.Although the strategies based on reducing the roles of humans and cattle as resources of infection decreased the infection rate and intensity quickly,re-infection occurred frequently due to the spread of snails and numerous animal reservoirs.Chemotherapy alone could not interrupt the transmission.By changing the traditional bio-medical control model,applying integrated control strategy mainly with social measures,integrating disease control with local social and economic development programs,changing the traditional life styles and agricultural production patterns,and eliminating the risk factors for schistosmiasis transmission,schistosomiasis control can be developed sustainably and reach the criteria of transmission interruption finally.

Objective:To explore the intramuscular nerve effect on the recovery of lacerated muscle. Methods:thirty (30) healthy New Zealand rabbits were involved in the study and randomly divided into groups A and B.The right musculus rectus femoris of the animal was chosen as the lacerated skeletal muscle model with the contralateral muscle as sham control. Group A underwent muscle laceration with the intramuscular nerve microsurgical repaired and muscle repaired;group B only with muscle repaired. After a postoperative period of 28weeks,measured the muscle wet weight and the muscle strength of musculus rectus femoris by electrical stimulation test and observed the change of morphology by Hematoxyline/eosin stain,Masson stain,NADH-TR stain and mATPase stain. Compared the difference of two groups through t-test and ANOVA to decide the effects of intramuscular nerve effect on muscle injury and repair. Results:At 28 weeks postoperative,Compared with the controls,wet weight and isometric contraction of musculus rectus femoris reduced in the intramuscular neurotmesis group (P

ObjectiveTo investigate the isolation effect of influenza virus by recombinant MDCK cells(MTY6 cells)stably expressing trypsinogen.MethodsAccording to the virus isolation method recommended by the World Health Organization(WHO)Global Influenza Surveillance Network(GISN)and the National Influenza Centers(NICs),a total of 20 throat swab specimens containing positive nucleic acid for H1N1,H3N2 and B influenza virus were isolated simultaneously using MDCK and MTY6 cells. Guinea pig red blood cells and chicken red blood cells were used for agglutination test respectively and the agglutination effects of different types of red blood cells,the positive rate of virus and the titer of hemagglutinin isolated from different cells were statistically compared.ResultsThe agglutination effect of the same virus isolate on the two types of red blood cells was different. The complete agglutination time of guinea pig red blood cells was about 2 times that of chicken red blood cells,and the deposition shape showed a ring shape. The average hemag-glutinin titer was 23. 6 ± 1. 2times that of chicken red blood cells. Under the same conditions,3 samples were negative for both types of cells,11 samples were positive for both types of cells,and the other 6 samples were negative for MDCK cells while positive for MTY6 cells. The positive rate of MTY6 cells was 30% higher than that of MDCK cells. The isolated positive samples included 8 cases of H1N1 subtype and the hemagglutinin titer of virus isolated by MTY6 cells was significantly higher than that by MDCK cells[13. 0(1. 7,23. 0)times on average]. 2 cases of H3N2 and 2 cases of B were isolated,the hemagglutinin titer of each virus isolated by MTY6 cells was 11. 3 and 32. 0 times higher than that by MDCK cells on average respectively.ConclusionIn conclusion,guinea pig red blood cells were superior to chicken red blood cells for influenza virus detection by cell isolation. Under the same conditions,MTY6 cells were more sensitive than MDCK cells for influenza virus isolation,and had the potential to be used as a high-quality cell matrix for influenza virus isolation.

AIM:To determine whether imperatorin would enhance the effect of doxorubicin therapy on cervi -cal cancer in vitro.METHODS:The viability of HeLa cells treated with imperatorin and doxorubicin was determined by MTT assay in vitro.The expression of Bcl-2 protein family ( Mcl-1, Bcl-2, Bcl-xL, Bad and Bax ) in HeLa cells treated with imperatorin and doxorubicin was evaluated by Western blot analysis .The apoptosis and mitochondrial membrane poten-tial (ΔΨm ) in the HeLa cells treated with imperatorin and doxorubicin were analyzed by flow cytometry .A Mcl-1 expres-sion vector was constructed , and its role in the cytotoxicity of imperatorin plus doxorubicin to HeLa cells was detected by MTT assay.RESULTS: Addition of imperatorin significantly enhanced the cytotoxicity of doxorubicin to HeLa cells in vitro.Mcl-1 expression was down-regulated by imperatorin but was not influenced by doxorubicin in the HeLa cells .A com-bination of imperatorin and doxorubicin induced apoptosis and ΔΨm collapse more significantly compared with the treatment with imperatorin or doxorubicin alone .Furthermore, the imperatorin-induced sensitization for doxorubicin cytotoxicity to HeLa cells was abolished by the transfection with Mcl-1 expression plasmid .CONCLUSION:The combination of doxoru-bicin with imperatorin enhances the antitumor effect of doxorubicin on cervical cancer cells via targeting Mcl -1.

Objective To investigate the association between diabetes and pancreatic cancer to provide the basis for the early di‐agnosis and prevention of pancreatic cancer .Methods By formulating the retrieval stategy ,the databases of PubMed ,EMbase ,Ov‐id 、EBSCO ,CNKI and CBMdisc ,VIP ,Wanfang/database etc .were retrieve by computer from their establishment to July of 2013 .At the same time ,the relevant literature references were also retrieved .All case‐control studies(CCS) on the association between diabe‐tes and pancreatic cancer were included and their quality was assessed .The meta analysis was conducted by using Review Manager 5 .0 software .Results A total of 21 CSC were included ,finally 18 case‐control studies met the inclusion criteria for conducting meta analysis ,including 6 184 cases in the pancreatic cancer group and 10 296 cases in the control group .The meta analysis on the pan‐creatic cancer group and the control group showed that the result of heterogeneity test from all research data was I2 = 74% ,χ2 =65 .38 ,P< 0 .01 ,the random effects model was adopted ,the pooled odds ratio(OR) was 3 .60 and 95% confidence interval(95% CI) was 3 .28 - 3 .94 ,the difference showed the statistic significance(P< 0 .05) ,which indicating that the risk suffering diabetes in the patients with pancreatic cancer was 3 .6 times of the normal population .Conclusion Diabetes is associated with pancreatic cancer and diabetes may be an early clinical manifestation of pancreatic cancer .

Biliary atresia(BA) is a destructive inflammatory obliterative cholangiopathy of neonates affecting both intrahepatic and extrahepatic bile ducts.BA is more common in east Asia.Genetic susceptibility,viral infections,and immune dysregulation may be related to BA,but immune dysregulation may play a key hole in the pathogenesis of BA.A current view of the pathogenesis of BA is that it may involve both a primary perinatal hepatobiliary viral infection and a secondary generation of an autoimmune-mediated bile duct epithelial injury.The etiology of BA is unknown,but there is evidence for the involvement of immunologic dysregulation mechanisms,which will be discussed in this review.

This article introduces the development history of ultrasound-assisted thrombolysis technology and operating points.It rainly focuses on the research status and application prospects of the technology in the treatment of acute cerebral infarction.

Computer science is not important enough in the medical university curriculum and students do not like this course. Therefore, game-teaching methods were employed to enrich teaching methods,to make medical teaching more vivid and to promote medical students' learning interests. In this paper,application of game-learning idea in the medical teaching process was discussed from multiple aspects including the story of the game,incentives of the game and the punitive measures of the game.