1.The Influence of Two Resins on Synthesizing HCV Polypeptides by Solid-phase Method
Li-Shu SUN ; Hong-Kun TANG ; Wu-Ling CHEN ;
China Biotechnology 2006;0(02):-
The solid-phase method to synthesis HCV polypetides was used.The connecting rate and the polypeptides' productivity and purity were compared when Wang resin and 2-chlorotrity choloride resin acting as vectors.By the identification of mass spectrum,the products' molecular weight was analysed.By ninhydrin method we compared to the connecting rate between two resins and amino acid.By RP-HPLC we carried out the analysis and purity of hetero peptides.The result showed the first amino acid's connecting rate along with the purity and productivity which we need when 2-chlorotrityl choloride resin acted as vector was notably higher than those when Wang resin acted as vector.So 2-chlorotrityl choloride as vector is used to synthesize HCV polypeptides,which adapts to the demand of big scale's industrialization production.
2.The function of Farnesol in formation of Staphylococcus epidermidis-Candida albicans mixed species biofilms
Xiaoyan WANG ; Kun YANG ; Yunchao HUANG ; Fengli GUO ; Ying CHEN ; Yujie LEI ; Qi TANG ; Xin LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(2):153-158
(P < 0.05 ).Scanning electron microscopy showed that the three-dimensional structure of mix-biofilm in control group was more complex.As real-time PCR showed,after 6-hour culture,the expressions of icaA,fbe,aap,hwp1, als3 and efg1 genes in Farnesol group were down-regulated compared to those in control group and after 24-hour culture the expressions of aap,hwp1,als3 and efg1 genes in Farnesol group were down-regulated compared to those in control group.Conclusion With the intervention by fungal quorum sensing molecules Farnesol,the structure in control group became denser and more complex than that in Farnesol group.This change may have a closer correlation with the down-regulated expressions of als3,hwp1 and efg1 genes,which are related to the formation of Candida albicans biofilms.
3.Efficiency of two different operation ways for a complex kind of retinal detachment
Heng, LI ; Fang, CHEN ; Wen-Jun, GOU ; Bi-Fang, TANG ; Quan-Kun, LIU
International Eye Science 2016;16(8):1579-1581
Abstract? AIM: To evaluate the efficiency of two different operation ways, vitrectomy and vitrectomy combined with inferior scleral buckling, for a complex kind of retinal detachment.?METHODS:The complex kind of retinal detachment were diagnosed in 100 cases ( 100 eyes ) , with the common features:1)the course of more than 1mo;2) at least one retinal hole located in the inferior marginal retina;3) the detachment of retina was found proliferated, there was at least 1 retinal fold in retinal detachment area. The patients were randomly divided into two groups: the treatment group ( 50 eyes ) who received vitrectomy combined with inferior scleral buckling;the control group ( 50 eyes ) who received vitrectomy without scleral buckling. The retinal anatomic reattachment, visual function recovery, macular central fovea thickness and complications in two groups were observed.? RESULTS: There were 49 eyes ( 98%) with retinal anatomic reattachment in the treatment group while 42 eyes(84%)in control group(χ2=4.2605, P<0.05);the best corrected visual acuity ( BCVA) was 0.254 ±0.238 in the treatment group while it was 0.219±0.229 in control group (t=1.8879,P>0.05);the macular central fovea thickness was 272 ±32.21μm in the treat group while it was 316 ± 33.46μm(t=12.597,P<0.01).Intraocular pressure in 12 eyes ( 24%) was more than 30mmHg in the treatment group while 4 eyes ( 8%) in control group within 1wk postoperation (χ2=4.7619,P<0.05); intraocular pressure in 100 eyes were controlled below 21mmHg 1mo postoperatively.? CONCLUSION: It is a more effective method to vitrectomy combined with inferior scleral buckling than the single vitrectomy for the special kind of retinal detachment.It can increase the rate of retinal anatomic reattachment and reduce macular edema.
4.Influence of up-regulated expression of HSP70 on myocardial KCa3.1 in rabbits with atrial fibrillation caused by rapid atrial pacing
Wei SONG ; Huikang CUI ; Yushui ZHENG ; Zhenling CHEN ; Xin XU ; Liang TANG ; Kun ZHU
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(6):606-609,610
Objective:To observe influence of up‐regulated expression of myocardial heat shock protein (HSP) 70 in‐duced by heat stress on myocardial calcium‐activated potassium channel (KCa ) 3.1 expression in rabbits with atrial fibrillation (AF) caused by rapid atrial pacing (RAP) .Methods :A total of 24 New Zealand white rabbits were ran‐domly divided into sham operation group (n=8 ,only implant electrode without pacing ) ,pacing group (n=8 ,right atrium (RA) received RAP at 600 times/min for 6h) and heat stress pacing group (heat stress group ,n=8 ,received heat stress preconditioning ,then the same RAP as pacing group ) .Results:Compared with sham operation group and pacing group ,there were significant up‐regulation of HSP70 mRNA and protein expression in different sites of heart [HSP70 protein ,left atrium (LA):(39.00 ± 3.21) vs .(39.75 ± 2.82) vs .(69.75 ± 3.45) ,RA: (38.38 ± 2.92) vs .(39.50 ± 3.89) vs .(69.00 ± 2.93) ,left atrial appendage (LAA):(37.75 ± 3.28) vs .(39.00 ± 3.89) vs . (68.63 ± 3.23) ,right atrial appendage (RAA): (37.00 ± 3.85) vs .(38.38 ± 3.74) vs .(68.75 ± 2.82)] in heat stress group , P<0. 01 all ,but there were no significant difference between pacing group and sham operation group , P>0.05 ;compared with pacing group with down‐regulation of KCa3.1 mRNA and protein expressions ,there were significant up‐regulation of KCa3.1 mRNA and protein expressions in different sites of heart [KCa3.1 protein ,LA:(21.25 ± 1.67) vs .(24.00 ± 2.62) ,RA :(21.13 ± 1.96) vs .(23.75 ± 1.83) ,LAA :(21.00 ± 2.07) vs .(23.75 ± 1.67) ,RAA:(20.88 ± 2.03) vs .(23.50 ± 2.45)] in heat stress group ,P<0.05 all ,and there were no significant difference between heat stress group and sham operation group , P>0. 05. Conclusion:Heat stress may induce up‐regulated expression of myocardial HSP 70 of myocardium ,and HSP 70 may inhibit down‐regulation of KCa 3. 1 mR‐NA and protein expressions in rabbits with atrial fibrillation.
5.Constructing a p53-fused dual luciferase reporter and verifying its function
Jing TAN ; Xianzhen JIANG ; Leye HE ; Zhiqiang JIANG ; Yuxin TANG ; Kun YAO ; Binghai CHEN ; Juan XUE
Journal of Central South University(Medical Sciences) 2013;38(11):1135-1140
Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.
6.Operative treatment of bi-column acetabular fractures
Yuqiang SUN ; Dongxu JIN ; Mingjie TANG ; Kun BAO ; Shengbao CHEN ; Bingfang ZENG
Chinese Journal of Orthopaedic Trauma 2008;10(12):1116-1120
Objective To probe into the operative techniques and clinical outcomes of treating bi-columnar acetabular fractures. Methods From April 2001 to December 2006, 609 cases undergoing operation for aeetabular fractures were followed up. Of them, 193 eases, 147 males and 46 females with an average age of 34.4 years, were of bi-columnar type. Their complications involved the articular cartilage of the femoral head in 37, eranioeerebral trauma in 31, injury to bladder and/or urethra in 27, injury to thorax and/or abdomen in 68, injury to pelvis and/or sacroiliac joint in 59, and injury to sciatic nerve in 11. The interval between injury and surgery was within 1 week in 15 cases, within 2 weeks in 121, within 3 weeks in 34, and beyond 3 weeks in 23. A single ilioinguinal approach was adopted for 4 cases and combined ap-proaches for 189. Results The mean operating time was 238 (150 to 330) minutes, and the blood loss averaged 1453 (450 to 4400) mL. The mean follow-up period was 44.2 (14 to 84) months. All the eases were evaluated by Matta reduction criteria, X-ray manifestations and the modified Merle d'Aubingne and Postel clinical scoring. One hundred and sixty-eight patients got anatomical reduction, 17 unsatisfactory reduction, and 8 poor reduction. The X-ray manifestations were excellent in 162 patients, good in 16, fair in 8, and poor in 7. The clinic results were excellent in 152 patients, good in 27, fair in 9, and poor in 5. The Kendall coefficient correlation between reduction and clinical outcome and that between reduction and X-ray manifes-tation were 0.74 and 0.77, respectively. Ectopic ossification happened in 77 eases and transient post-operative sciatic nerve paralysis occurred in 3. Conclusions The combined approaches facilitate exposure and synergetie reduction of the bi-colunmar fractures of acetabulum. Reduction of the acetabular roof is essential to restoration of the normal contour of the acetabulum. Reduction bears a positive correlation to the clinical results as well as experience of surgeons.
7.A meta-analysis of factors influencing the development of gastric cancer in Chinese populations
Dandan YANG ; Xuecheng YAO ; Xinhan ZHANG ; Mengling TANG ; Jianbing WANG ; Mingjuan JIN ; Kun CHEN
Journal of Preventive Medicine 2022;34(6):561-570
Objective:
To investigate the factors influencing the development of gastric cancer in Chinese populations, so as provide insights into creating a model for predicting gastric cancer incidence among Chinese populations.
Methods:
The case-control and cohort studies pertaining to factors affecting the development of gastric cancer were retrieved in electronic Chinese and English databases, including CNKI, Wanfang Data, VIP, PubMed, Web of Science and Embase from their inception until September 30, 2021. A meta-analysis was performed using R package version 4.1.0. Sensitivity analysis was performed using the “leave-one-out” evaluation procedure, and the publication bias was evaluated using the Egger regression test and the trim-and-fill procedure.
Results:
A total of 5 301 publications were screened and 116 eligible studies were included in the final analysis, including 103 case-control studies and 13 cohort studies, which covered approximately 3.23 million study subjects. A total of 45 factors affecting the development of gastric cancer were collected, and there were less than 4 publications reporting 7 factors, which were only qualitatively described. There were 38 factors included in the final meta-analysis. A total of 21 factors were identified as risk factors of gastric cancer, including a history of gastrointestinal diseases (pooled OR=4.85, 95%CI: 3.74-6.29), H. pylori infection (pooled OR=3.18, 95%CI: 2.35-4.32), binge eating and drinking (pooled OR=2.88, 95%CI: 2.09-3.97) and a family history of tumors (pooled OR=2.78, 95%CI: 2.17-3.56), and 10 factors as protective factors, including vegetable intake (pooled OR=0.48, 95%CI: 0.38-0.61), tea consumption (pooled OR=0.55, 95%CI: 0.47-0.64), administration of aspirin (pooled OR=0.53, 95%CI: 0.31-0.92) and administration of statins (pooled OR=0.59, 95%CI: 0.44-0.80). Sensitivity analyses of eating moldy food frequently, white meat intake, favoring spicy food and administration of sulfonylureas were not robust. Following correction with the trim-and-fill procedure, there was still a publication bias pertaining to high income, diabetes, administration of stains, alcohol consumption, tea consumption and white meat intake.
Conclusions
The development of gastric cancer is associated with a medical history of gastrointestinal disease, H. pylori infection, family history of tumors and poor dietary habits. Risk and protective factors of gastric cancer are recommended to be included in models used to predict gastric cancer incidence among Chinese populations.
8.Study on the complex hepatitis C virus antigen with multi-genotype E1 epitopes
Feng HE ; Xiaoguo SONG ; Li TANG ; Guohua WANG ; Heqiu ZHANG ; Kun CHEN ; Cuixia ZHU ; Libo XU ; Shigan LING
Chinese Journal of Microbiology and Immunology 2009;29(1):46-52
Objective To design a complex hepatitis C vires(HCV)=E1 antigen,and to search its application in HCV vaccine and diagnosis test.Methods Through consulting the database and widely comparison of sequences from HCV E1 of different genotype,the representative immunodominant epitope sequences were selected from all the six genotypes.Their genes were deduced according to the preference codon in E.coli and three fragments were designed to contain the six epitopes.They were chemically synthesized and cloned into pBVIL1 vector separately.The cloned fragments were conjugated together profiting from pBVIL1's property,and then a doubled pan-DR helper T cell epitopes(PADRE)gene was inserted to form a complex expression plasmid.The transformed E.coli cells with this plasmid were cultured and induced to express recombinant protein and the antigenic activity of the product was tested.Results An expressing plasmid containing 8 epitopes from HCV genotype 1a,1b,2a,3a,4a,6a and a doubled PADRE sequences was constructed successfully and the engineering E.coli transformed with this plasmid highly expressed after inducing culture at 42℃ in an inclusion manner.The immunological activity of the purified recombinsnt multi-epitope antigen shows that:(1)It can react with a great part of sera from HCV positive patients by indirect ELISA.(2)It can induce notable specific humoral immunity in injected mice.Conclusion The novel constructed expressing plasmid and its product may be useful in study of a newly HCV vaccine as well as an antigen for HCV immunoassays.
9.Study on occupational exposure limit of dimethoxymethane in workplace air.
Hao CHEN ; Lin LIN ; Mao-tang ZHANG ; Qu LIU ; Gang LI ; Bi-kun YU ; Li CUI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(10):787-791
OBJECTIVETo study the occupational exposure limit of dimethoxymethane (DMM) in workplace air.
METHODSLiterature research was conducted on DMM; information on its physicochemical properties, toxicology, and occupational epidemiology was collected. Upon analyzing these data, an occupational exposure limit of DMM in workplace air was proposed according to the guidelines and principles of developing occupational exposure limits for harmful factors in workplaces.
RESULTSA total of 165 air samples were collected from DMM manufacturers, 14 of which had a DMM concentration over 3000 mg/m(3), accounting for 8.48% of all samples. Compared with the control group, the exposed employees had a significantly increased incidence of discomfort symptoms when exposed to more than 6000 mg/m(3) DMN (P < 0.01).No significant differences in the constituent ratios of abnormal results in physical examinations were found between the exposure groups and control group (P > 0.05).
CONCLUSIONWe recommend to set the permissible concentration-time weighed average to 3100 mg/m(3) as the occupational exposure limit of DMM in workplace air in China. This occupational exposure limit standard meets the relevant requirements of toxicology and occupational epidemiology, as well as fits well with our country's current economical and technological levels. This standard has high applicability in China's domestic settings.
Air Pollutants, Occupational ; analysis ; China ; Humans ; Inhalation Exposure ; Methyl Ethers ; analysis ; Occupational Exposure ; analysis ; Threshold Limit Values ; Workplace
10.Application of mutated inbred animal models in the experimental teaching of medical genetics
qing Zuo TANG ; yun Xue HUO ; ying Yu HAN ; Jing WANG ; hong Hai YE ; Tian ZHAO ; kun Zhen LI ; wen Zhen CHEN
Chinese Journal of Comparative Medicine 2017;27(10):123-126
Objective Mutated inbred animal model is introduced to the practical course of genetic diagnosis in the hope that medical students are able to apply what they have learned to clinical cases, based on a deep understanding of principle and technology on gene mutation detection. Methods We integrated DNA extraction, polymerase chain reaction, agarose gel electrophoresis, and gel imaging analysis into a comprehensive experiment and arranged 4-year-programme undergraduates majoring in preclinical medical sciences to conduct it with the purpose of investigating the internal relations between phenotype and genotype in a hairless Uncv mouse model. Subsequently, the questionnaire aimed at evaluating learning effect on the part of students was handed out and their feedbacks were analyzed. Results More than 90% of respondents are satisfied with the general learning effect. Especially, 98. 7% of students support the enhancing effect of the new teaching mode on their research skills and 96% consider the practical course helpful to their problem-solving ability. Conclusions The introduction of mutated inbred animal model to the practical system of molecular diagnostics proves beneficial to boost students' learning effect and scientific research quality. Our practice also provokes thoughts on the further utilization of animal models in teaching system of medical sciences.