3.Gene Coloning,Expression and Enzymatic Assay of Human sPLA2-IIA
Xiu-Fei YIN ; Yu-Hua LIAO ; Jia-Quan WANG ;
China Biotechnology 2006;0(08):-
Objective: To clone the cDNA of human sPLA2-IIA,construct the engineered Escherischia coli expressing human sPLA2-IIA and identify the expressed human sPLA2-IIA. Methods: Total RNAs were purified from human fetal spleen. The cDNA of human sPLA2-IIA was cloned by RT-PCR and inserted into plasmid pET32a(+) between NcoI and EcoRI sites for expressing the recombinant human sPLA2-IIA in Escherischia coli BL21(DE3). The recombinants were screened by SDS-PAGE. The engineered Escherischia coli expressing trxA-human sPLA2-IIA fusion protein was established. The expressed human sPLA2-IIA exists in the form of inclusion body and accounts for about 25% of the total proteins of Escherischia coli BL21(DE3). Conclusion: the engineered E. coli methods are suitable for preparing plenty of human sPLA2-IIA which has laid base for the large-scale expression,purification and basic studies of human sPLA2-IIA.
4.Effect of nanometer and micron silicon dioxide dusts on oxidation and anti-oxidation index in mice.
Su-hua WANG ; Mao-lin DU ; Dian-quan ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2009;27(10):637-638
Animals
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Dust
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Male
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Malondialdehyde
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blood
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Mice
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Nanoparticles
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Oxidation-Reduction
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drug effects
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Particle Size
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Silicon Dioxide
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toxicity
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Superoxide Dismutase
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blood
5.Clinical efficacy on vertebrobasilar insufficiency treated with auricular acupuncture.
Zun-Hua SHU ; Guo-Qiang WANG ; Ming-Quan LI ; Xiu-Ge WANG
Chinese Acupuncture & Moxibustion 2014;34(12):1161-1164
OBJECTIVETo compare the efficacy on vertebrobasilar insufficiency (VBI) between auricular acupuncture therapy and oral administration of medicine.
METHODSSixty patients of VBI were randomized into an auricular acupuncture therapy group and a medicine group, 30 cases in each one. In the auricular acupuncture group, acupuncture was applied bilaterally to gan (CO12) and jiejie (HX8) on the ears and needles were retained for 15 min. After needle withdrawal, the vaccariae semen were fixed with plaster at naogan (AT3, 4i), zhen (AT3), jing (AH12), shen (CO10) and pi (CO13) on the ears. In the medicine group, flunarizine hydrochloride capsules (Sibelium), 5mg were prescribed for oral administration, once every night. The treatment lasted continuously for 2 weeks (14 days) in the two groups. In 2 weeks, the clinical efficacy was assessed and the transcranial doppler (TCD) examination was performed.
RESULTSAfter treatment, the symptom scores were all apparently reduced in the patients of the two groups (P < 0.01, P < 0.05). Compared with the medicine group, the reduced score was much more obvious in the auricular acupuncture group (P < 0.05), indicating the significant difference. After treatment, with TCD examination, the blood velocity was increased to different degrees in the patients of low velocity type in the auricular acupuncture group and the medicine group; that was reduced to different degrees in the patients of high velocity type in the auricular acupuncture group and the medicine group. All of them were different significantly as compared with those before treatment (all P < 0.05). But the difference was not significant between the two groups (both P > 0.05). In comparison of clinical efficacy between the two groups, the effective rate was 93.3% (28/30) in the acupuncture group and better than 76.7% (23/30) in the medicine group, indicating the significant difference in comparison (P < 0.05).
CONCLUSIONThe auricular acupuncture therapy achieves the definite efficacy on VBI and the efficacy is better than flunarizine hydrochloride capsules.
Acupuncture Points ; Acupuncture, Ear ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Vertebrobasilar Insufficiency ; therapy
7.Screening of the pORF3 binding protein of HEV in human liver cells by using yeast double hybridization system
Yanjun KANG ; Quan SHEN ; Wen ZHANG ; Xiuguo HUA ; Xueqin RAN ; Jiafu WANG
Chinese Journal of Zoonoses 2010;(2):111-114
HEV is a positive strand RNA virus containing 3 open reading frames(ORFs), while the function of its binding protein pORF3 is poorly understood. In our study, the interactive protein of the pORF3 from human liver cDNA library was screened by the SOS recruitment system, and the construction of the pSOS bait-ORF3 plasmid was identified. The expression of the pORF3 was analyzed by Western blotting, and the detection result of the self activation of bait protein was obtained as expected. Totally, 8 putative interacting proteins had been successively screened. Some of these cellular binding proteins of pORF3, especially the important immunological pathway kinase p110δ, may advance our understanding of the role in ORF3 protein of HEV, and the possible implications of these candidate proteins for the further research are discussed.
8.Changes in serum interleukin-1 beta,interleukin-6,and tumor necrosis factor alpha in patients with lumbar disc herniation after Shenshi Tongluo therapy:Randomized controlled study
Jie YU ; Li-Guo ZHU ; Jing-Hua GAO ; Shang-Quan WANG ;
Chinese Journal of Tissue Engineering Research 2007;0(07):-
AIM:To observe the changes in serum cytokines in patients with lumbar disc herniation(LDH) following Shenshi Tongluo therapy(discharging damp and dredging collaterals).METHODS:①Forty-three patients with LDH(cold-damp arthralgia type) were selected from Wangjing Hospital,China Academy of Chinese Medical Sciences from February to December 2005,and randomly divided into experimental group(n =22) and control group(n =21).The informed consent was obtained from all subjects.Meanwhile,30 healthy volunteers were selected as controls.②The patients in the experimental group orally took modified Semen Coicis decoction(Semen Coicis,eucommia bark,Radix dipsaci,cocculi,Chinese clematis root,caulis millettiae,angelica root,achyranthis,and paeoniae radix) according to Shenshi Tongluo therapy,one dose daily for 4 weeks.The patients in control group took Yaotong Ning capsule,4 pills per day,once daily for 4 weeks.③ Contents of serum interleukin-1 ?(IL-1?),IL-6,tumor necrosis factor-?(TNF-?) in patients were detected using radioimmunoassay before and after treatment.Clinical outcome was evaluated using visual analog scale(VAS) and assessment criteria for low lumbar pain of Japanese Orthopedic Association(JOA).RESULTS:①Contents of serum IL-1?,IL-6,and TNF-? in patients were higher than healthy people(P
9.The Extraction Method of Bacterial DNA from Activated Sludges
Wei ZHENG ; Chun-Shan QUAN ; Yong-Zhe PIAO ; Jun-Hua WANG ; Sheng-Di FAN ;
China Biotechnology 2006;0(07):-
Methods for studying the population diversity of microorganism in activated sludge usually require enrichment of bacterial genome.The efficient information on microbial species composition provided and shifted in diversity revealed are dependent on the effective DNA recovery technique.The method was based on washing by alkaline phosphate buffer and digestion with extended heating of the activated sludge suspension in the presence of lysozyme and freeze-thawing in high-salt-SDS buffer.The extraction was tested for four activated sludge differing in places and dates.The DNA fragment from all sludge was integrity.DNA yields ranged from 105 to 823 ?g/g sludge and were of sufficient purity for PCR-based 16S ribosomal DNA analysis and restriction digested.In general,all methods produced DNA pure were not enough for PCR amplification and libraries construction.As basis of experimental goals,the study provides an appropriate extraction method of microbial DNA in sludge.
10.Proliferation and identification of dendritic cells from peripheral blood of patients with bladder cancer in vitro
Dan CAI ; Zhi-Hua WANG ; Zhi-Quan HU ; Xu ZHANG ; Si-Wei ZHOU ; Zhang-Qun YE
Chinese Journal of Urology 2001;0(07):-
Objective To investigate the proliferation and identification of dendritic cells(DC)de- rived from peripheral blood of patients with bladder cancer in vitro.Methods The mononuclear cells were prepared from peripheral blood of patients with bladder cancer by Ficoll-Hypaque centrifugation method,and were induced by the recombinant cytokines hGM-CSF(50 ng/ml),hlL-4(10 ng/ml)and hTNF-?(50 ng/ ml)for 2 weeks.The growth and morphology of DC were observed through the phase contrast or electron mi- croscope,and their pheuotypes were determined by flow cytometry.The capacity of DC to activate T cell-de- pendent anti-tumor immune responses was tested by MTT method.Results The DC cultured in vitro turned into suspensive growth from adhesive situation on the 6th day,then the number of DC increased con- tinuously and the cells showed the irregular morphologic appearance of DC with veiled edges on the 8th day. Flow cytometry showed that the mature DC expressed high levels of specific markers such as CD_(1a),CD_(83), CD_(86)and HLA-DR.T cells activated by DC showed strong cytotoxicity to bladder cancer cell line BIU87 with a killing rate of(48.8?3.7)%,while the killing rate of T cells which were not activated by DC was(25.7?1.5)%;the difference of the rate between them was significant(P<0.01). Conclusions The DC can be cultured from peripheral blood of patients with bladder cancer by induction of rhGM-CSF,rhIL-4 and hT- NF-?in vitro.This may lay an experimental foundation for further research on DC vaccine.