2.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.
3.Transcatheter arterial chemoembolization and delayed surgery for infant hepatoblastoma
xian-liang, HU ; wen-lv, SHEN ; jian-hong, LI ; xue-wu, JIANG ; zhong-xian, CHEN
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To evaluate the effect of transcatheter arterial chemoembolization (TACE) and delayed surgery for infant hepatoblastoma.Methods TACE was performed with the initial digital subtractive angiography (DSA) under general anesthesia 1-3 times in 8 infants with huge hepatoblastoma, whose age was 2 to 12 months. DSA was done via arterials in hepatoblastoma each time before chemoembolization. The arterials were perfused with chemodrugs and suspensions in ultrasome iodized oil , and were blocked with spring rings. DSA findings indicated that the tumor shrank without new tumorous arterials after 1 month in 6 cases, and 4 of them showed no tumorous staining, and the delayed surgery was performed successfully 1 week later in 6 infants. One boy underwent systemic chemotherapy alone during 6 months after 3 times of TACE. Results TACE therapy did not encounter any major technical problem or toxic reaction caused by chemotherapy. The following DSA test 4 weeks later did not detect any new tumorous vessels in 6 cases. Six children received TACE and surgery had been followed-up with no tumor recurrence for months averagely. The boy underwent TACE and venous chemotherapy for 6 months , without surgery , had been followed-up for 48 months until the present report. CT, AFP and DSA did not show any hints of tumor recurrence. Six cases receiving 3 times TACE combined with surgery survived without tumor recurrence. Conclusions TACE is a very effective, safe and helpful therapy for hepatoblastoma, which stressed the repeated use of spring ring to block tumor vessels lastingly if necessary. If surgery is required, DSA test is needed beforehand to detect new tumorous vessels or neoplasm. If there is any , TACE is repeated. TACE combined with surgery may provide an additional promising choice in the treatment of hepatoblastoma, and repeated TACE alone may cure hepatoblastoma in infants.
4.Exercise based on traditional Chinese medicine for patients with stable chronic obstructive pulmonary disease
Wen ZHANG ; Xian-Qiao JIN ; Wen-Hua CHEN ; Wei-Qing WU ; Rong YANG ; Yan-Yan YU ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(12):-
Objective To make up an exercise prescription based on traditional Chinese medical training (EP-TCMT) for patients with stable chronic obstructive pulmonary disease (COPD).Methods Eighty-five pa- tients with stable COPD were randomly divided into a control group (CG group),a traditional Chinese medicine group ( TC group) and an exercise prescription group ( EP group).The patients in the TC and EP groups were giv- en intensive training for 8 weeks.Their 6 rain walk distance (6MWD) and Borg scale scores were assessed before and after the treatment.Results The 6MWD in the TC group increased from 337.68?59.18 m to 386.14?76.71 m,while those in the EP group improved from 348.00?55.94 m to 425.17?53.22 m.The Borg scale scores in the TC group decreased from 3.14?1.94 to 2.32?1.25,while those in the EP group declined from 3.45?1.84 to 1.72?0.70.Conclusion Making up EP-TCMTs is feasible.Additional treatment was found to improve exercise tolerance and decrease dyspnea in COPD patients.Exercise therapy based on traditional Chinese methods is easy and safe.
5.Contrast Research on Decreasing Rate of Serum ?_2-Microglobulin before and after Children′s Hemodialysis and Hemodiafiltation
cui-hua, LI ; jiang-wei, LUAN ; yan-xiang, WU ; xing-xian, YANG ; xiao-wen, WANG
Journal of Applied Clinical Pediatrics 2006;0(18):-
Objective To explore the influence of hemodialysis(HD) and henodiafiltation on serum ?_2-microglobulin(?_2-MG) of children with acute renal failure and contrast research on declining rate of serum ?_2-MG before and after(children′s) HD and HDF.(Met-)hods By Branc Dialog HD machine of double-pump,HDF for 18 times and HD for 20 times were given to children with acute renal(fai-)lure.The serum ?_2-MG were observed before and after HD and HDF.Results A great difference was observed in level of serum ?_2-MG between before and after HD and HDF.In HDF group,there was significant difference in level of serum ?_2-MG before and after HDF(P
6.Study on Mutation in Exon 8 of ATP7B Gene in Chinese Patients with Wilson Disease
xian-ting, JIAO ; xiao-qing, LIU ; ya-fen, ZHANG ; jie, WU ; lian-wen, WANG
Journal of Applied Clinical Pediatrics 2004;0(08):-
G,the noval insertion mutation of c.2298_2299insC is identified in Chinese patients.
7.Cloning and expression of the hy1 gene of Enterococcus faecium and immunologenicity of the Hy1 fusion protein
Li-Xian WU ; Wen-Xiang HUANG ; Tao LUO ; Xiao-Yang JIAO ;
Chinese Journal of Infectious Diseases 2007;0(10):-
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.
8.The quality of life and its influential factors in patients with 3 major rheumatic diseases
Shao-Xian HU ; Wen-Ze XIAO ; Fang KONG ; Jin-He XIONG ; Jin MAO ; Ji-Ping WU ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(10):-
Objective To explore the quality of life (QOL) and its influential factors among patients with 3 major rheumatic diseases. Methods A total of 216 patients with rheumatic diseases (84 patients with systemic lu- pus erythematosus, SLE, 83 with rheumatoid arthritis, RA, and 49 with ankylosing spondylitis, AS) were recruited. The information with regard to their quality of life, sociopsychological factors and the evaluation of disease activity were obtained by using the medical outcomes study short form-36 (SF-36) and clinic documents. Results Patients with rheumatic diseases scored significantly lower with each subscale of SF-36 as compared to those of a healthy popu- lation in China (P
9.FERMENTATION CONDITIONS OF SOYMILK-CLOTTING ENZYME PRODUCTION BY BACILLUS SP.AND STUDIES ON THE PROPERTIES OF THE ENZYM E
Yong-Mei JIANG ; Wen-Xian ZHANG ; Qiao-Qin SHI ; Song-Gang WU ;
Microbiology 1992;0(04):-
The optimum condition of shaking-flask p roducing enzyme were the tempe rature 26℃,initial pH 6 4,fermentation period 19 hours,medium volume 15mL m e dium/300mL Flask.soymilk-clotting enzyme was obtained from ammonium sulfate p r ecipitation.The optimum temperature and pH for the soymilk-clotting activity wa s 70℃and 5 8.The enzyme was easy to lose activity in acid or alkaline circumst a nce.About 60% of the original activity remained after 1 hour at 60℃.Ca 2+ ,Fe 2+ , Mg 2+ ,Na +increased the clotting activity,whereas Zn 2+ ,Al 3+ ca use inhibition.
10.Optimization of expression and purification of recombinant Salvia miltiorrhiza WRKY1 protein in Escherichia coli.
Yu-Zhong LIU ; Ye SHEN ; Qi-Xian RONG ; Wen-Yan WU ; Rui-Bo LI ; Zhi-Gang WU ; Min CHEN
China Journal of Chinese Materia Medica 2014;39(7):1214-1219
WRKY transcription factor is one of the Zinc finger proteins which contains a highly conserved WRKY domain and is a family of the plant-specific transcription factor. The plasmid pET28a-SmWRKY1 harboring Salvia miltiorrhiza WRKY1 (SmWRKY1) gene was successfully transformed and expressed in Escherichia coli BL21 (DE3). The conditions on protein expression of SmWRKY1 in E. coli, including induction duration, temperature, IPTG concentration and the E. coli concentration were optimized. The results showed that the highest protein expression of SmWRKY1 was obtained at 24 hours after the E. coli was cultured in the presence of 0.2 mol x L(-1) IPTG at 20 degrees C with A600 values of 1.0-1.5. This recombinant histidine-tagged protein was expressed at 2.454 g x L(-1) as inclusion body, which was first extracted using urea, and then purified by Ni2+ affinity chromatography and identified by SDS-PAGE. The expression of SmWRKY1 in E. coli was further confirmed by western blotting analysis.
Blotting, Western
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Cloning, Molecular
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DNA-Binding Proteins
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chemistry
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genetics
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isolation & purification
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metabolism
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli
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chemistry
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genetics
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metabolism
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Molecular Weight
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Plant Proteins
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chemistry
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genetics
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isolation & purification
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metabolism
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Recombinant Proteins
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chemistry
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genetics
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isolation & purification
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metabolism
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Salvia miltiorrhiza
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genetics