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.

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

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.

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.

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.

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.

Objective To systematically evaluate the value of narrow band imaging(NBI)and iodine staining in early diagnosis of esophageal cancer and precancerous lesions.Methods The databases of PubMed,Embase,CNKI,VIP technological journals and Wanfang digital journals were retrieved by computer for searching the literatures on the value of NBI and iodine staining in diagnosing early esophageal cancer and precancerous lesions.The Meta-Disc 1.4 software was used to conduct the meta analysis and calculate the pooled sensitivity,specificity and 95％ confidence interval(95％ CI).Summary receiver operating characteristic(SROC)curve was drawn and the area under curve(AUC)was calculated,thus the diagnostic values of the two methods were evaluated.Results Totally 13 articles were included,including 975 patients.The pooled sensitivity and specificity for NBI were 0.88(95％CI=0.86 to 0.90)and 0.79(95％CI=0.76 to 0.82).The pooled sensitivity and specificity for iodine staining were 0.95(95％CI=0.93 to 0.97)and 0.45(95％CI=0.40 to 0.49).AUC of SROC for the two methods were 0.938 6 and 0.952 9 respectively.There was no statistically difference in the AUC values between the two methods(Z=0.519,P>0.05).Conclusion NBI and iodine staining are both effective ways in the diagnosis of early esophageal cancer and precancerous lesions.And both of them have a certain clinical value.

Objective To study pharmacodynamic changes of combined anesthesia with remifentanil at different target plasma concentrations and isoflurane at different minimal alveolar concentrations(MAC) in laparoscopic operations. Methods Forty-five patients with ASA status Ⅰ~Ⅱ,receiving selective laparoscopic cholecystectomy or laparoscopic oophorocystectomy,were included.The patients were randomly divided into three groups with 15 patients in each group.In the Group A,anesthesia was maintained with isoflurane inhalation at the MAC of 1.3;in the Group B,anesthesia was maintained with both isoflurane inhalation at the MAC of 0.6 and remifentanil target-controlled infusion at 4 ng/L;and in the Group C,anesthesia was maintained with both isoflurane inhalation at the MAC of 0.4 and remifentanil target-controlled infusion at 6 ng/L.Pharmacodynamic parameters and recovery characteristics,including bispectral index(BIS),heart rate variability(HRV),mean arterial pressure(MAP),heart rate(HR),time to additional administration of muscle relaxants,time to the recovery of spontaneous breathing,time to eye opening,time to endotracheal extubation,time to the recovery of orientation,and follow-up survey of "awareness during operation",were compared among the 3 groups.Results ①The BIS value was lower in the Group A than in the Group B and C at the time of pneumoperitoneum for 2 min and removal of the gallbladder or the ovarian cyst.The measures of HRV were higher in the Group A than in the Group B and C at the time of pneumoperitoneum for 2 min and removal of the gallbladder or the ovarian cyst.The measures of MAP and HR in the Group A were higher than those in the Group C at the time of pneumoperitoneum for 2 min,and were lower than those in the Group C at the time of the endotracheal extubation and the recovery of consciousness.②Either isoflurane inhalation at the MAC of 0.6 combined with remifentanil target-controlled infusion at 4 ng/L or isoflurane inhalation at the MAC of 0.4 combined with remifentanil target-controlled infusion at 6 ng/L provided satisfactory anesthetic maintenance.③Significantly shorter time to endotracheal extubation,to eye opening,and to the recovery of orientation were recorded in the Group B and C than in the Group A.Conclusions Anesthesia with target-controlled infusion of remifentanil combined with inhalation of isoflurane can be used for laparoscopic surgery.Pharmarcodynamic parameters and recovery characteristics show that combined use of remifentanil infusion at 4 ng/L and isoflurane inhalation at the MAC of 0.6 provides optimal anesthetic maintenance.

Anaerobic threshold (AT) and maximal oxygen uptake (Vo_2max) were determined according to ventilation and gas exchange measurements during graded cycling in 26 athletes and were compared with cardiac condition and function measured by 2-dimensional echocardiography exercise test in corresponding with HR at and above AT. There are significant correlations between Dd and VO_2max and between cardiac function (such as SV, CO, UMO and so on) and VO_2, but the correlations are higher in the same than in different subjects of the same HR. It suggests that the physiological individualities including cardiac function and other peripheral factors are so different that the VO_2max, AT, cardiac condition and function should be measured respectively to evaluate aerobic capabilities of each athlete, thus guiding the training and selection of athletes.