Objective To investigate the clinical significance of H2A histone family member(H2AX)gene in human lung adenocarcinoma,and to explore its biological role in lung adenocarcinoma.Methods The tran-scriptome data of lung adenocarcinoma were downloaded from the Cancer Genome Atlas and Gene Expression database.The expression of H2AX mRNA in lung adenocarcinoma tissues and adjacent normal lung tissues was extracted by R 4.3.1 software,and the relationship between the expression of H2AX and the clinicopath-ological parameters of the patients was analyzed.Kaplan-Meier and Cox regression analysis were used to ana-lyze the prognostic value of H2AX gene expression.The expression of H2AX gene in lung adenocarcinoma cell lines and normal bronchial epithelial cells was verified by quantitative real-time PCR(qPCR).Gene set enrich-ment analysis,gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were used to explore the biological role of H2AX gene.Results The expression of H2AX mRNA in lung adenocarcinoma tissues was significantly higher than that in adjacent normal tissues(P<0.001).qPCR results showed that compared with normal bronchial epithelial cells,the expression levels of H2AX mRNA in lung adenocarcinoma cell lines A549 and SPC-A1 were significantly increased(P<0.05).H2AX expression was associated with tumor stage(P=0.029)and N stage(P=0.016).Kaplan-Meier curve analysis showed that high expression of H2AX was associated with a lower overall survival rate of patients(P<0.001).Cox regression analysis showed that H2AX was an independent prognostic factor for lung adenocarcinoma patients(HR=1.011,95%CI 1.003-1.020,P=0.006).The results of functional enrichment analysis showed that H2AX gene was involved in molecular functions such as cell cycle,DNA replication,mismatch repair,and P53 signaling pathway.Conclusion H2AX gene is highly expressed in human lung adenocarcinoma,which is relat-ed to the malignant progression of tumor patients.H2AX gene can affect the progression of lung adenocarcino-ma through a variety of pathways,and may be used as a molecular marker for lung adenocarcinoma patients.

Objective To investigate the relationship between Helicobacter pylori (HP) infection and the development of chronic urticaria.Methods Published case-control studies which concerned HP infection related chronic urticaria were searched in Wanfang,CNKI,CQVIP Chinese databanks and PubMed.Meta-analysis was applied to analyze the pooled odds ratio (OR) and 95％ confidence interval (CI).Results 37 studies which comprised 2 909 cases of chronic urticaria and 1 873 persons served as controls were enrolled.When compared with the controls,HP infection significantly increased the risk of chronic urticaria development with a pooled OR of 3.20 (95％CI:2.31-4.43).Results from Meta-regression analyses showed that the distribution of residential areas and detection method being used were potential influential factors.Conclusion HP infection seemed to be associated with an increased risk of developing the chronic urticaria.

Objective To investigate the expression and significance of γH2AX in cervical squamous carcinoma.Methods Firstly,DNA were extracted from 74 cervical squamous carcinoma samples and PCR were tested for HPV infection.Secondly,formalin-fixed paraffin-embedded tissue sections (4 μm)were stained with H&E method to detect cervical lesions grading.Thirdly,HPV16 DNA were examined by in situ hybridization(ISH) and γH2AX,p16 were examined by immunohistochemical (IHC) staining.Then,30 cases typical tissue sections in which including the normal cervical tissue,cervical intraepithelial neoplasia and cervical carcinoma in situ were selected for comparing the HPV DNA loading,and the γH2AX and,pl6 expression.Finally,the feasibility of γH2AX serving as a biomarks in HPV infection-related cervical carcinogenesis were analyzed.Results In this study,HPV infection ratio is 98.65％,and HPV16 is the most common type with 74.32％ infection.In situ hybridization showed no HPV16 DNA exist in normal cervical tissues and CINI.In CIN Ⅱ HPV DNA exist mainly as episomal DNA.With the increasing of cervical lesions grade,HPV DNA was integrated into chromosome steadily.The expression of γH2AX and pl6 were positively associated with grading of cervical lesions.HPV DNA and γH2AX protein co-exist primarily in the prickle cell layer and the granular cell layer.The HPV DNA and p16 protein exist in different cell layer.Conclusion γH2AX may be employed as a biomarker for HPV positive cervical carcinogenesis.

This study was aimed to identify pET21b-HPV16E2/BL21(DE3) strain and to optimize the expression of human papillomavirus type 16 (HPV16) E2 protein by orthogonal analysis. Four influence factors on two levels were selected to increase the target protein quantity. The four factors were induction time, induction temperature, inductor concentration and cell density. The quantity of HPV16 E2 protein was used as the evaluation parameter. Induced by IPTG, HPV16 E2 protein was analyzed by SDS-PAGE and Western Blot. Target protein was analyzed by GIS imaging system to quantify the protein level. SPSS13. 0 software was applied to analyze the result. Data showed that the expression strain pET211rHPV16 E2/BL21(DE3) was identified correctly. HPV16 E2 protein expressed mainly at insoluble form. The 42KD protein band was identified by SDS-PAGE and Western blot. Orthogonal test was applied on influence factor analysis and expression optimization successfully. Main influence factors were inductor concentration and induction temperature. The optimimum condition of maximum expression quantity was 37 degrees C, 7h, 1.0 mmol/L IPTG and OD600 1.0. In this experiment, orthogonal test could not only be used to analyze the influential factors and promote the target protein expression, but also be used to provide a better experiment method for molecular biological study.
DNA-Binding Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; metabolism ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomavirus Infections ; virology ; Recombinant Proteins ; biosynthesis ; genetics

0.05).Genotype B with YMDD mutations in 10 samples were found,including 2 YIDD(M+I) mutations and 8 YVDD(M+V) mutations.Genotype C with YMDD mutations in 68 samples were found,including 44 mutations I(M+I) and 24 mutations V(M+V).There was significant difference in YMDD mutation types between genotypes B and C(?2=5.5,P

This study was intended to establish a method of purification of HPV16 L1 protein expressed in a prokaryotic system and to obtain the purified protein. The prokaryotic expression vector pGEX-4T-1-HPV16 L1 was constructed and transformed into E. coli BL21 cell, and induced by 1 mM IPTG to express HPV16L1 protein. The inclusion bodies were isolated and solubilized with 8 M urea. After the urea was removed by gradual dialysis, the denatured L1 protein were renatured and then were purified by affinity chromatography. The results showed that HPV16L1 protein formed inclusion bodies in bacterial expression system, suggesting that this assay can be used to purify HPV16L1 protein and hence provide a basis for studying the applications of HPV16 L1 protein.
Capsid Proteins ; biosynthesis ; Escherichia coli ; metabolism ; Genetic Vectors ; Human papillomavirus 16 ; Oncogene Proteins, Viral ; biosynthesis

Objective To evaluate a new gene therapy for the treatment of experimental occlusive arterial disease Methods Magnetic nanospheres were produced, VEGF gene was cloned for subsequent construction of eukaryotic expression plasmid The magnetic gelatin microspheres used in targeted gene therapy were prepared by emulsion crosslinking method The microspheres were injected intrafemorally in rabbits through contralateral femoral artery, and the ischemic limb was placed in a magnetic field Angiography was performed on day 10 and day 30 respectively The capillary density and the capillary to muscle fiber ratio were determined histochemically Results Compared to the controls there was significant collateral artery development in VEGF transfected group The capillary density and the capillary to muscle fiber ratio were significantly higher for the VEGF transfected group than for the control group. The capillary density of control was (125?23)/mm 2, and in VEGF group was (298?27)/mm 2, P

Objective:To study the effect of the cytokines, tumor necrosis factor-?(TNF-?), interferon-?(IFN-?), interferon-?(IFN-?), platelet-derived growth factor BB(PDGF-BB), basic fibroblast growth factor(bFGF),on promoter activity of human transforming growth factor-?1(TGF-?1) gene.Methods:A construct of phTGF2.14 containing sequence from -1 328 bp to +812 bp of human TGF-?1 gene,linking with chloramphenicol acetyltransferase(CAT) as reporter gene,was transiently transfected into rat hepatic stellate cell line nFSC. The cells were subsequently treated with TNF-?,IFN-?,IFN-?,PDGF-BB,bFGF, and the CAT activity was assessed 48 hours after stimulation with each cytokines.Results:TNF-? of 10 ng/ml can increased the CAT activity of phTGF2.14 to 3.24 fold compared to control. IFN-? and IFN-? at 1 000 U/ml decreased CAT activity to (42?12)% and (58?6)% of control respectively.PDGF-BB,bFGF of 10 ng/ml had no effect on promoter activity of human TGF-?1 gene;Combined application of IFN-?,IFN-? and TNF-?,the promoter TGF-?1 gene were 1.32 and 1.46 fold compared with control,respectively.Conclusion:These data indicate that TNF-? can increase the promoter activity of human TGF-?1 gene, but IFN-?,IFN? can downregulate the CAT activites of phTGF2.14, and IFN-?,IFN-? can interdict the upregulate effect of TNF-? on phTGF2.14. We did not find PDGF-BB,bFGF have any effect on TGF-?1 promoter. These provided an essential evidence for study the interaction mechanism of cytokines in fibrogenisis diseases.

Measuring the length of the proportional units (PU) of different truncal parts in males and females and analyzing the relationship between these measurements showed that there was nosignificant gender difference in the length of one cun respectively for the perpendicular proportional units (PPU) of prothorax, epigastrium and flank but there were significant differences in the PPU of the hypogastrium, lateral thorax and hypochondrium and the horizontal proportional unit (HPU) of the back; within the same gender, there was no significant difference in the length ofone cun between the PPU of the prothorax and that of epigastrium and no significant difference betweenthe PPU of the flank and the HPU of the intermammillary region in males but there was a significant difference between PU of other parts.

Measuring the length of the proportional units of the lower limb and analyzing their interrelations showed that between genders there were significant differences in a length of one can of perpendicular proportional unit for various regions of the lower limb; within the same gender, there were different results in comparing the lengths of one of perpendicular proportional unit among various regions of the lower limb.