As human beings ascend to high altitude, a number of reactions may occur against hypoxic injuries. These hypoxic responses are related to intake, transportation and utility of the oxygen. As a crucial subcellular organelle of oxygen utility, mitochondrion is a central link of high altitude acclimatization, adaptation and mountain sicknesses. In this review, we discussed the recent advances in researches on hypoxic mitochondrial responses at high altitude.
Adaptation, Physiological ; Altitude ; Altitude Sickness ; Animals ; Humans ; Hypoxia ; Mitochondria ; pathology ; Oxygen ; physiology

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

Objective To construct the serine protease gene(sprE)mutant and to study the pathogenicity of sprE gene of Enterococcus faecalis.Methods Recombinant suicide vector pCQ001 of Enterococcus faecalis with pTX4577,was constructed.Then,created isogenic sprE-deficient mu- tant(*sprE)by allelic replacement was constructed.Moreover,the growth ability and the virulence of the mutant were compared with those of the wide type in vitro and in vivo respectively.A mouse peritonitis model and a rabbit endocarditis model were utilized in the study.Results The *sprE was selected by kanamycin and identified by polymerase chain reaction(PCR),pulsed field gel electropho- resis(PFGE)and Southern blot.The evidences showed that the sprE gene had a major role in helping bacteria to resist the elevated temperature and oxidative stress.The virulence of mutant decreased af- ter sprE gene was knocked out.Conclusions The *sprE of Enterococcus faecalis is constructed suc- cessfully,sprE gene is important in the pathogenesis of Enterococcus faecalis,which probably is a major virulence factor of Enterococcus faecali.

Objective To investigate the effect of video-based teaching in the training of nursing operation for newly-contracted nurses. Methods One hundred and twenty one nurses newly recruited in September 2011 to September 2012 were set as the control group, another 128 in October 2012 to October 2013 were set as the experiment group. The former were trained and assessed with traditional training method and the latter were trained for 1 year in video-based teaching methodology. After training, both groups were examined about their operation skills and meanwhile a survey on video-based teaching was conducted. Result After training, the results in operating skills evaluation in the experiment group were significantly better those of the control group (Z=2.82, P<0.05). Conclusion Compared with traditional training method, the video-based teaching can raise the level of nursing operation skills and significantly improve the quality of nursing, thus worthy of popularization and application.

Objective: Changes in the ?2 integrin of adhesion molecules between cells are closely associated with the invasion and migration of tumor cells.This study aimed at the effect of anti-?2 integrin on the invasion and migration of osteosarcoma MG63 cells. Methods: The osteosarcoma MG63 cell line was cultured in the DMEM medium.The effect of the anti-?2 integrin monoclonal antibody on the migration and invasion of tumor cells were measured by scratch assay and Transwell assay.The migration and invasion cells were stained by Crystal violet staining and counted under the hundredfold microscope.Results: Compared with the control group,the migration and invasion abilities of the MG63 cells were significantly decreased in the anti-?2 treatment groups.Conclusion: Anti-?2 integrin may inhibit the migration and invasion of osteosarcoma cells.

Three hundred and twenty-one bacteria strains were obtained from rice leaves,stem,root tissue and paddy field soil,of which the number of strains which can inhibit mycelium of Magnaporthe grisea growth markedly was fifty-seven through fermentation in 2.0 mL Eppendorf tube,and among these fifty-seven strains,five strains were strongly antagonistic to Magnaporthe grisea.These five strains was identified for their morphologic,physiological and biochemical characteristics,and the results showed that one strain(No.156)was bacillus subtilis,two strains(No.171 and No.177)were Bacillus pumillus and two strains(No.192 and No.279)were Bacillus ploymyxa.

Objective To construct a recombinant expressing plasmid of the hy1 gene of Enterococcus faecium and to express the recombinant Hy1 protein in E.coil.To explore the immune response in mice fed orally with Hyl protein.Methods hy1 gene was amplified by polymerase chain reaction(PCR)and inserted into a prokaryotic expression vector pQE-30.The recomhinant plasmids were transfected into DH5_?to express Hy1 fusion proteins,which were purified by Ni~--column. Western blot was employed to confirm the immunogenicity of the purified protein.Mice were immu- nized by feeding with the fusion protein.The concentrations of antigen-specific antibody in the serum, mucosal fluid and faces were detected by enzyme-linked immunosorbent assay(ELISA).The role of these antibodies in the anti-infection response was evaluated after the mice were challenged with TX0016.Results hy1 gene was sequenced as 1662 bp,the fusion protein encoding polypeptides of 553 amino acid residues.The relative molecular weight was 60 000 when it was determined by sodium dodecylsulfatepo-lyacry-lamide gel electropboresis(SDS-PAGE).The dissolvable expression protein accounted for 38% of total cell protein.After processed by affinity chromatography,the purity of fusion protein was above 92%.Western blot analysis confirmed that fusion protein could be specifically recognized by the anti-TX0016 serum.The concentrations of serum IgA,serum IgG,faeces sIgA and intestmucosal fluid sIgA was 0.365?0.048,0.431?0.064,0.743?0.056 and 1.112?0.113 respectively in hy1 groups and 0.051?0.013,0.098?0.019,0.102?0.032 and 0.187?0.051 respectively in control group.The differences were statistically significant.The mice survival rate after TX0016 challenge was 70% in hyl group and 50% in control group.There was significant difference between these two groups.Conclusion The results indicate that oral immunization with hyl can induce effective mueosal immune response and produce high level sIgA.

Objective To explore the expressions of Survivin and VEGF and relationship between them in hepatocellular carcinoma(HCC).Methods The expressions of Survivin protein and VEGF protein in 50 HCC.30 cirrhosis and 10 normal tissues were assessed by immunohistochemical method.The expressions of Survivin mRNA and VEGF mRNA in 50 HCC,30 cirrhosis and 10 normal tissues were assessed by in situ hybridization.Results The expressions of Survivin and VEGF in cancer tissues,cirrhosis tissues,normal tissues weresignificantly different. The expression of Survivin in HCC tissues was stronger than that in cirrhosis,but the expreesion of VEGF in cirrho- sis was stronger than that in HCC tissues.Conclusion The expression of survivin.is closely associated with the ex- pression of VEGF in HCC and they take positive correlation.The abnormal expressions of Survivin and VEGF are closely associated with the development of HCC.They may play important roles in the development of HCC.