2.Effect of VIM in EV71 infection in human microvascular endothelial cells
Yanyun ZHONG ; Bao ZHANG ; Mingliang HE ; Yujuan CAO ; Xianbo WU
The Journal of Practical Medicine 2016;32(10):1560-1563
Objectives To study the effect of VIM in Enterovirus 71 (EV71) infection of (human brain microvascular endothelial cells (HBMEC) and elaborating the mechanism of EV71 infection in the nervous system. Methods Knocked out the VIM by CRISPR technology , the differences in EV71 absorption , replication , release between wild VIM and VIM knocked-out (VIM-KO) HBMEC were detected by fluorescence quantitative PCR. Results 4 ℃ absorption experiment conformed that EV71 adsorption in VIM- KO is 40% less than in the normal HBMEC. After EV71 infect HBMEC for 48 h (48 h p. i.), the quantitative PCR result showed intracellular viral RNA in VIM-KO was only 1/12 of that in the normal HBMEC. Also the extracellular viral RNA was quantified, and the number of cells in VIM-KO had been reduced 1.4 times compared with the normal HBMEC. Conclusions Once VIM knocking out, EV71 attachment has been obviously reduced. Meanwhile, the level of viral RNA replication and release are decreased compared with the normal HBMEC. VIM may be an attachment receptor of EV71 in HBMEC , when the virus invades HBMEC with the binding of VIM. Moreover , VIM plays an important role in the replication and release of EV71.
3.Overview of CDISC standard and implementation in China.
Victor WU ; Wen-Jun BAO ; John WANG ; Rui-Ling PENG ; Ya-Zhong DENG ; Zi-Bao ZHANG
Acta Pharmaceutica Sinica 2015;50(11):1428-1433
CDISC standard has become a set of global data standards that can be used in clinical study, covering the full life cycle of clinical researches. After nearly 20 years of development and continuous version upgrades, CDISC standard can improve the quality and efficiency of clinical research and drug review, and to facilitate all stakeholders involved in researches to exchange the study data and communicate the outcomes. CDISC standard has been or is to be adopted as standard format in data submission by multiple regulatory authorities, and more widely implemented by the global pharmaceutical community. CDISC standard is gradually adopted in China. The feasibility and roadmap of CDISC standard as the Chinese data submission format requirements are undergoing exploration and piloting further.
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4.Surgical treatment of the tumors of posterior part of third ventricle and pineal region
Liang, GUAN ; Bao-min, ZHANG ; Wei-Jian, SUN ; Wei-zhong, YANG ; Bao-guo, LIU ; FU-yu, WANG
Bulletin of The Academy of Military Medical Sciences 2001;25(1):54-56
Objective:To summarize the experiences of the surgical treatment of tumors in the posterior part of the third ventricle and pineal region. Methods: Twenty-seven patients with tumors in this region treated microsurgically from 1990 to 2000 were analyzed. The surgical indications, approaches, and operative key points were discussed. The prognostic factors were also analyzed. Results:Tumors were removed totally in 11 patients, removed subtotally in 7 and removed partially in 5. Biopsy and ventriculo-peritoneal shunt was performed in 4 patients. Of 17 patients in follow-up, 13 patients survived longer than 5 years. Conclusions: Most tumors in the posterior part of the third ventricle and pineal region can be surgically removed.Sufficient specimen obtained in the operation can confirm pathologic property of the tumor,guiding next chemotherapy and radiotherapy.
5.Eukaryotic expression and DNA immunization of hemolysin HlyX gene from Leptospira serovar Lai
Bi HUANG ; Lang BAO ; Qi ZHONG ; Zhengling SHANG ; Huidong ZHANG ; Ying ZHANG
Chinese Journal of Microbiology and Immunology 2008;28(2):134-138
Objective To study on the expression of the eukaryotic recombinant vector carrying HlyX gene of Leptospira serovar Lai in mammalian cell and explore the humoral immune response in BALB/c mice immunized with the recombinant plasmid. Methods The HlyX gene was amplified from Leptospira serovar Lai genomic DNA by PCR and inserted into pcDNA3.1 vector. After transformed into E. coli DH5α,the recombinant plasmid was assayed for identification by PCR analysis,restriction nuclease enzyme digestion and sequencing. The recombinant plasmid was transfected into COS-7 cells,then RT-PCR and Western blot were performed to test the expression of the target gene. The recombinant plasmid was injected intramuscularly into BALB/c mice for three times at intervals of two weeks,and the antibody titer was measured by ELISA. Results PCR showed the full length HlyX gene was about 1100 bp. PCR analysis,restriction nuclease enzyme digestion and sequencing indicated the recombinant vector was constructed successfully. After the plasmid Was transfected into COS-7 cells,a fragment about 1100 bp was found by RT-PCR and a specific band relative molecular mass(Mr)about 40×103,which was consistent with the expected size of the target proteins was showed by Western blot. ELISA showed the antibody titer in BALB/c mice immunized by the HlyX gene of Leptospira serovar Lai can elicit high-titer antibody in BALB/c mice,which has laid the foundation for the application of the DNA vaccine.
6.Effect of lumbar nerve dorsal roots section on the rat bone structural changes of lower limb
Feng XU ; Zhong-Guo FU ; Dian-Ying ZHANG ; Ya-Ming CHU ; Bao-Guo JIANG ;
Chinese Journal of Microsurgery 2000;0(03):-
Objective To investigate the rat bone structural changes of lower limb following lumbar nerve dorsal roots section.Methods Forty-eight mature female Wistar rats were divided into posterior radi- cotomy(PR)and comtrol groups randomly.The bilateral femoral bone mineral density(BMD)and biome- chanics characteristics were analyzed 2,4 and 8 weeks after the radicotomy.The same operation except the radicotomy was done in the sham group.Results In PR group,2,4,and 8 weeks after the radicotomy,the BMD of femur was(0.221?0.008)g/cm~3,(0.213?0.015)g/cm~3 ,and(0.216?0.105)g/cm~3 ,respective- ly;while that was(0.223?0.005)g/cm~3,(0.218?0.014)g/cm~3 ,and(0.208?0.111)g/cm~3 in control group.No significant difference was observed between the two groups(P>0.05).In PR group,2,4,and 8 weeks after the operation,the mean maximum load in three-point bending test of femun midshaft was(93.64?8.76)N,(89.77?11.18)N and(93.21?8.74)N,respectively,and was lower than the values of the con- trol group at the same time point(95.94?6.29)N,(91.63?9.43)N,(95.57?8.64)N,However,there was no significant difference between the two groups(P>0.05).Accordingly,there was no significant difference in the energy absorption in femun midshaft between the two groups(P>0.05).Conclusion The selective rhizotomies of part lumbar never dorsal roots might not cause the loss of the femur BMD and the change of bio- mechanics property significantly in short period.
7.Recombinant human interleukin-6 inhibits mRNA expressions of adiponectin,its receptor 1 and adiponectin secretion in SW872 adipocytes
Long-Jiang ZHANG ; Han-Hua LIN ; Hong-Wei WANG ; Zhong-Xian BAO ; Xiong-Hui XIAN ;
Chinese Journal of Endocrinology and Metabolism 2001;0(05):-
SW872 preadipocytes were cultured and induced to differentiate by oleic acid in vitro.The levels of adiponectin and its receptors (AdipoR1 and AdipoR2) mRNA were measured by semiquantitative RT- PCR.The concentration of adiponectin in the culture medium was assayed by ELISA.The results showed that rhIL- 6 could inhibit adiponectin,AdipoR1 mRNA expressions and adiponectin secretion in SW872 adipocytes in a time- and dose-dependent manner,but did not influence adipoR2 mRNA expression.
8.Initial study of quantitative analysis of fatty liver by ~1H-MR spectroscopy imaging
Chang-Hong LIANG ; Yu-Bao LIU ; Zhong-Lin ZHANG ; Shu-Fei XIE ; Qiu-Shi WANG ;
Chinese Journal of Radiology 2001;0(01):-
Objective To investigate the feasibility of ~1H-MR spectroscopy(~1H-MRS)imaging to quantitatively detect fatty liver.Methods Twenty patients with fatty liver and 11 healthy volunteers underwent plain CT scan,conventional MR imaging and ~1H-MRS analysis.The blood lipid and liver function were tested on the same day as the MR examination.~1H-MRS sequence measured the peaks of H_2O and lipid,and the areas under the peaks.The relative contents of the lipid compound were calculated,and compared with the results of CT scan and liver function tests.Results The CT values of the normal group and the fatty liver group were(59?9)HU and(24?11)HU respectively.On ~1H-MRS a protruding high H_2O peak and a flat low lipid peak were observed in the normal group,while the protruding high H_2O peak and a high lipid peak appeared in the fatty liver group.The values of lipid peak in the normal group and the fatty liver group were(0.05?0.01)?10~5,(0.70?0.24)?10~5 respectively(t=4.32,P0.05),the areas under the lipid peak were(1.36?0.73)?10~9、(2.35?1.15)?10~9 respectively(t=5.21,P0.05).Conclusion ~1 H-MRS imaging is feasible to quantitatively detect liver fat and is a non-invasive method for detecting early fatty liver.
9.Overexpression of Alcohol DehydrogenaseⅠ in Saccharomyces cerevisiae
Li-Na QIN ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.
10.Cloning and Expression of Acyl Carrier Protein Gene from Schizochytrium
Zhi-Ping LI ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.