1.Pathophysiological changes in mitochondria of mammalian exposed to hypoxia at high altitude.
Wen-xiang GAO ; Gang WU ; Yu-qi GAO
Chinese Journal of Applied Physiology 2014;30(6):502-505
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
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Altitude
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Altitude Sickness
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Animals
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Humans
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Hypoxia
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Mitochondria
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pathology
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Oxygen
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physiology
2.Prokaryotic expression and purification of human Smith D1 antigen
Wen-Bing WU ; Xiao-Peng LAN ; Xiang-Yue YANG ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
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.
3.Eukaryotic Expression and Primarily Application of Human Smith D1 Antigen in Methylotrophic Yeast Pichia pastoris
Xiang-Yue YANG ; Wen-Bing WU ; Xiao-Peng LAN ;
Microbiology 1992;0(01):-
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.
4.Application of video-based teaching in the training of nursing operation skills for newly-contracted nurses
Xiang PENG ; Wen ZHOU ; Guojv SONG ; Wenyan WANG ; Hanjun WU
Modern Clinical Nursing 2015;14(10):54-57
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.
5.Anti-?2 integrin inhibits invasion and migration of osteosarcoma MG63 cells
Wen GUO ; Xiang LU ; Xin SHI ; Sujia WU
Journal of Medical Postgraduates 2003;0(12):-
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.
6.Construction of serine protease gene mutant and study on pathogenicity of serine protease gene of Entero-coccus faecalis
Li-Xian WU ; Wen HUANG ; Sun XIANG ; Ping XIAO ;
Chinese Journal of Infectious Diseases 2000;0(02):-
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.
8.The investigation of risk factors of post-polypectomy emergency hemorrhage
Yadi ZHANG ; Wensheng PAN ; Hong SHEN ; Wanjun ZHANG ; Liangqin WU ; Wen HU ; Yimiao ZHU ; Xiang XU
Chinese Journal of Emergency Medicine 2012;21(1):74-78
Objective To explore the risk factors in post-polypectomy hemorrhage in rectum and to discuss the appropriate interventions.Methods A total of 313 patients with 373 polypi were included in this study. The clinical data were analyzed by SPSS 16 software. Results There were 313 patients with colorectal polypus curatively resected and 373 polypi in total.There were 11 (3.5%) patients subjected to post-polypectomy hemorrhage in rectum.Regression analysis showed that the independent risk factor of postpolypectomy hemorrhage in rectum was the hypertension of patients (P < 0.01 ) and this hemorrhage had no significant correlations with patientg'ender,age,size of polypus,pathological characteristics and the methods of polypectomy.Conclusions Hypertension of patients is an independent risk factor in post-polypectomy hemorrhage.
9.Intermittent porta hepatis occlusion is a risk factor of postoperative intra-hepatic recurrence of hepatocellular carcinoma
Shenghua HAO ; Xiang-wu YANG ; Wen-long YANG ; Chi-dan WAN
Chinese Journal of Hepatobiliary Surgery 2013;(5):321-324
Objective To investigate the impact of intermittent porta hepatis occlusion on postoperative intra-hepatic recurrence of hepatocellular carcinoma.Methods We retrospectively reviewed the records of 335 patients who underwent partial hepatectomy.The patients were classified into 2 groups:(1) the study group (n=97):porta hepatis was occluded with intermittent Pringle maneuver with 2-3 cycles of clamp/unclamp time of 15 min/5 min,repeated 2-3cycles; (2) the control group (n=238):including using Pringle maneuver,preconditioning occlusion of porta hepatis and selective occlusion of portal blood inflow.Patients were followed-up in the Outpatient Department once every 2-3 weeks in the 1st year,and once every 3-6 weeks in the 2nd year with US/CT/MRI and serum AFP test.The mean duration of follow-up was 26.5 months.Results The perioperative mortality was 1.8% (6/335).Tumour recurrence in the study group was 31.6% and 48.4% in the 1st and 2nd year,respectively.The recurrence rates were significantly higher,than the 21.4% and 38.0% in the control group (P<0.05).To exclude the miscellaneous factors which were involved in intra-hepatic recurrence of HCC,we set up 3 criteria to include patients for subgroup analysis:tumor ≥5 cm; serum AFP decreased to normal level within 4 weeks; negative intra-operative US scan.The number of patients included were 79 and 155 in the study and the control groups,respectively.There were significant differences in recurrence rate between the study and the control groups in the 1st and 2nd year (29.1% vs 18.7%,46.8% vs 35.5%,P<0.05).There were no significant differences in overall survival rate between the two groups.Conclusions Intermittent porta hepatis occlusion is a risk factor of postoperative intra hepatic recurrence of hepatocellular carcinoma.
10.Expression and significance of peripheral blood T helper cells in human leucocyte antigen-B27-anterior uveitis patients
Wen-jun, ZOU ; Zhi-feng, WU ; Xiao-li, XIANG ; Song, SUN ; Jie, ZHANG
Chinese Journal of Experimental Ophthalmology 2012;(12):1118-1121
Background Human leucocyte antigen (HLA)-B27-associated uveitis is one of the most common causes of non-infectious uveitis.T helper 17 (Th17) cells play an important role in human autoimmune diseases,but the pathology research on the production of Th17 cells in acute anterior uveitis patients positive for HLA-B27 was rarely reported.Objective The aim of this study was to investigate the expression and significance of the peripheral blood T helper cell subsets (Th1,Th2,Th17) in acute anterior uveitis patients positive for HLA-B27.Methods This study meets the criteria of the Helsinki Declaration.Informed consent was obtained from all the participants.A prospective cohort design was used in this study.Twenty-two patients with acute anterior uveitis positive for HLA-B27 were enrolled from Affiliated Second Hospital of Nanjing Medical University,and 16 normal healthy subjects with matched gender and age were enrolled as controls.The expression of interferon-γ (IFN-γ),interleukin-4 (IL-4) and IL-17 on lymphocytes (CD4+) in blood were assessed by flow cytometry,and immunoturbidimetry was used to detect the C reactive protein (CRP) level in blood.The degree of the severity of disease was evaluated by clinical scoring.The correlations between the percentage of IFN-γ+Th1,IL-4+Th2,or IL-17+Th17 with clinical factors and CRP were analyzed.Results The percentages of IFN-γ+Th1 and IL-17+Th17 in the peripheral blood were (23.11 ±9.69) % and (3.96±2.92) % in the patient group,showing a significant increase in comparison with (16.00±4.26)% and (1.68±0.60) % in the control group (P=0.041,P=0.002).However,the IL-4+Th2 level was not significantly different between the patient group (0.33% ±0.36%) and the control group (0.56% ±0.34%) (P=0.122).No significant correlations were found between the percentage of IFN-γ+ Th1 with disease severity and CRP (r =0.197,P =0.500 ; r =0.253,P =0.383),between the percentage of IL-4+ Th2 with disease severity and CRP (r =0.068,P =0.817 ; r =0.439,P =0.116) as well as between the percentage of IL-17 + Th17 with CRP (r =0.226,P =0.436).However,a positive correlation was seen between the percentage of IL-17+ Th17 with disease severity (r =0.805,P =0.001).Conclusions IFN-γ and IL-17 in human CD4+T cells are significantly elevated in the blood of HLA-B27-related acute anterior uveitis patients.Disease severity is associated with the percentage of IL-17 +Th17,suggesting that Th1 cells together with Th17 cells participate in the pathogenesis of the disease and Th17 cells might play a dominant role in the disease.