Objective To study the effects E.coli purine nucleoside phosphorylase suicide gene on human gastric cancer MKN-45 cells as well as the bystander effect.Methods The ePNP gene was subcloned to pcDNA3.1 to construct recombinant eukaryotic expression vector pcDNA3.1-ePNP.The recombinant plasmid was transfected into MKN-45 cells by lipofectamin method and positive cell clones were screened with G418.Then MePdR was added to these gene-modified cells and the sensitivity of the cells to MePdR was studied as well as the bystander effect.Results MePdR treatment of MKN-45/ePNP cells induced cytotoxic and antiproliferative effects in a concentration-dependent manner with 100% cell death since 1 mg?L-1.Bystander effect was strong in vitro as 100% of tumor cells were killed by MePdR with only 10% MKN-45/ePNP cells.The gene-modified gastric cancer cells MKN-45/ePNP were sensitive to the treatment of MePdR compared with unmodified tumor cells,and also a remarkable bystander effect was observed.Conclusion The efficiency of ePNP/MePdR system and the potential feasibility of suicide gene strategy for clinical therapy of human gastric cancer are demonstrated.

Objective To clone human cell division cycle gene 2(CDC2)from human liver cancer tissue and express CDC2 protein in E.coli BL-21(DE3).Methods The total RNA was extracted from liver cancer tissue and amplified by reverse transcription polymerase chain reaction(RT-PCR).The PCR products were cloned into the prokaryotic expression vector pET28a(+)followed by DNA sequencing.The recombinant pET28a(+)/CDC2(rCDC2)plasmid was transformed into E.coli.The rCDC2 was induced with IPTG and characterized by SDS-PAGE.Results The cloned CDC2 gene was composed of 894 nucleotides,and is accordance with that reported in GenBank.The prokaryotic expression vector was constructed successfully.The CDC2 protein was successfully expressed in E.coli.Conclusion The CDC2 cDNA is successfully cloned from human liver cancer tissue,and can express in E.coli.

Objective:To investigate the in vitro effect of Bifidobacterium BB12 on IL-8 secretion by Caco-2 cells induced with S.enteritidis.Methods:The coculture systems of Caco-2 cells with S.enteritidis alone or combined with the strain of Bifidobacterium BB12 were established. The expression of IL-8 mRNA and NF-?B p65 in the cells were examined by reverse transcription PCR, real-time PCR and EMSA, and IL-8 in the supernatant was measured by ELISA.Results:In control group, there was little IL-8 mRNA expression in Caco-2 cells, NF-?B p65 was seldom in Caco-2 cells’ nuclei, and the concentration of IL-8 in the supernatant was as few as 126 ng/L. After stimulated by the S.enteritidis, the expression of IL-8 mRNA and activated NF-?B p65 was found in Caco-2 cells (P

Objective To study the relationship between activity and virulence of secretory acid proteinase and extracellular phosphlipase of different Candida albicans isolated from various specimens.Methods 190 strains of Candida albicans were isolated from various specimens(sputum,blood,thrush,wound,secretion of vaginitis) of patients with Candida albicans infection in hospital.Milk-plate medium and egg-yolk medium were used respectively to test the activities of secretory acid proteinase and extracellular phosphlipase of Candida albicans.The suspension of two forms of Candida albicans(5?106CFU?mL-1) were injected to vein of tail in mice respectively in virulence test.The mortality and mean survival time of mice were observed in 1 month.The virulence was appreciated with the mean survival time of mice.Results All of 190 strains,the positive detectable rates of secretory acid proteinase and extracellular phosphlipase were 83.68% and 85.26%,respectively.Animal experimental results showed the activities of secretory acid proteinase and extracellular phospolipase in hyphal forms were significantly higher than that in spore form(P

Objective To clone the rat heme oxygenase-1(RHO-1) from rat spleen and express RHO-1 in E.coli BL-21.Methods The total RNA was extracted from rat spleen and amplified by reverse transcription polymerase chain reaction(RT-PCR).PCR products were cloned into pMD18-T(TA)vector followed by DNA sequencing.RHO-1 cDNA fragments in TA vector were subcloned into the prokaryotic expression vector pET28a(+).The recombinant pET28a(+)/RHO-1(rRHO-1) plasmid was transformed into E.coli.The rRHO-1 was induced with IPTG and characterized by SDS-PAGE.Results The cloned RHO-1 gene was composed of 870 nucleotides,and was accordance with the sequence reported in GenBank.The prokaryotic expression vector was constructed successfully.The RHO-1 protein was successfully expressed in E.coli.Conclusion The prokaryotic expression vector of rRHO-1 has been constructed,and the fusion protein has been successfully expressed.

Objective To investigate the effects of 15-deoxy-?12,14-prostaglandin J2(15d-PGJ2) and DDP on the growth of human pulmonary carcinoma PLA-801D cells and the mechanisms of apoptosis.Methods The human pulmonary carcinoma PLA-801D cells were selected and added to each well of 96-well place and cultivated for 24 h.Then the cells were treated with different concentrations of 15d-PGJ2(0,5,10,20,40 and 80 ?g?L-1) or 15d-PGJ2 combined with DDP(3 mg?L) for 24 h.0 ?g?L-1 15d-PGJ2 group was control group.The morphological changes of cells were observed under inverted microscope.Microculture tetrazolium(MTT)dye was applied to detect the proliferation of the human pulmonary carcinoma PLA-801D cells treated with 15d-PGJ2 and DDP.Diphenylamine assay(DPA) was used to evaluate the activation.Flow cytometry assay(FCM) was used to detect the apoptosis proportion and the changes of cell cycle.Results When the human pulmonary carcinoma PLA-801D cells were treated with low-concentration 15d-PGJ2 alone(5,10 and 20 ?g?L-1),no significant difference was observed in the inhibitory rate of cell growth and the apoptotic indexes such as the apoptosis proportion,the percent of DNA fragmentation and the activity of caspase-3 compared with control group(P

Objective:To investigate the association of human leucocyte antigen-DRB1 alleles with chronic hepatitis B and hepatic cirrhosis in Han population of Jilin area.Methods:HLA-DRB1 alleles in 61 patients with chronic hepatitis B,44 patients with hepatic cirrhosis,32 spontaneous rehabilitees after HBV infectin,and 50 healthy controls were analyzed by using polymerase chain reaction/sequence specific primer technique.Results:The frequency of HLA-DRB1*1201-3 allele in chronic hepatitis B group was higher than that in healthy control group(17.21% vs.8.00%,P=0.0427,RR=2.391).The frequency of HLA-DRB1*0701 allele in chronic hepatitis B group and hepatic cirrhosis group was markedly higher than that in healthy controls group(11.48 % vs.2.00%,P=0.006 6,RR=6.35;12.50% vs.2.00%,P=0.004 6,RR=7.00)and in spontaneous rehabilitee group (11.48% vs.1.56%,P=0.0183,RR=8.17;12.50% vs.1.56%,P=0.0136,RR=9.00).No significant differences frequencies of the other HLA-DRB1 alleles were observed among the four groups.Conclusion:Alleles of HLA-DRB1*1201-3 and HLA-DRB1*0701 are closely associated with chronic hepatitis B and maybe susceptible genes or linkage genes of chronic hepatitis B.HLA-DRB1*0701 is also closely associated with hepatic cirrhosis and maybe a susceptible gene or linkage gene-posthepatitis B-cirrhosis in Han population of Jilin area.

Purpose:To study the expression level of bcl 2 gene in breast cancer tissues, paracancer tissues, metastatic lymph nodes and normal tissues. To study the relation of the gene expression and tumor formulation, development and clinical significance. Methods:Semi fixed quantity RT PCR was performed in 16 patients to detect the level of bcl 2 gene expression in cancer tissues, paracancer tissues, metastatic lymph nodes and normal tissues. Results:The relative values of mRNA in cancer tissues was 0.264?0.137 and 0.272?0.076 in metastatic lymph nodes and 0.131?0.083 in paracancer tissues were higher than those in normal tissue (0.033?0.031), so the difference was significant( P

BACKGROUNDLung cancer is one of the most common cancers worldwide and its mortality rate has increased year after year. Molecular biology has contributed to make people's understanding to the disease at the gene level. A new idea will be given to the early diagnosis and treatment for lung cancer with the method of molecular biology which can be used to find the association between some genes and lung cancer. The aim of this study is to evaluate the relationship between polymorphism of codon 72 (BstU I single nucleotide polymorphism, BstU I SNP) in p53 gene and susceptibility and radiosensitivity of non-small cell lung cancer (NSCLC) in Chinese population.

METHODSThe BstU I single nucleotide polymorphic sites at condon 72 in exon 4 of p53 gene in 50 patients with NSCLC as well as 50 healthy controls were inspected by polymerase chain reaction-restricted fragment length polymorphism assay and the relationship between BstU I SNP and susceptibility and radiosensitivity of NSCLC was analysed by case-control test.

RESULTSThe allelic distribution of the three genotypes (A1/A1, A1/A2, A2/A2) in healthy controls was 32.0%, 42.0% and 26.0% respectively, which differed slightly from that of lung cancer patients, which was 28.0%, 32.0% and 40.0%. These allelic and genotype differences between control and lung cancer groups (A1/A1 OR=0.83, 95% CI 0.36-1.27 and A2/A2 OR=1.90, 95% CI 0.82-4.42) were insignificant. The patients with A1/A1 or A2/A2 genotype were sensitive to radiotherapy and the patients with A1/A2 were not sensitive to radiother-apy (Chi-square=9.2, P < 0.05), the effective rate to radiotherapy were 71.4%, 70.0% and 25.0% respectively.

CONCLUSIONSThere is no significant relationship between the BstUI SNP in p53 and susceptibility in NSCLC .The p53 BstU I SNP is closely associated with radiosensitivity of NSCLC in the Northern Chinese population.

Objective:To evaluate whether posterior cruciate ligament (PCL) avulsion fracture was correlated with intercondylar notch and tibial slope.Methods:A total of 48 patients with PCL avulsion fracture were compared with 48 age- and sex-matched healthy controls without PCL avulsion fracture from January 2012 to October 2018 in our hospital. There were 25 males and 23 females with an average age of 54.35±14.06 years (range 27-82 years) in PCL avulsion fracture group. MRI of the knee joint were acquired in the sagittal, coronal, and axial sequences. The measurements included intercondylar notch's angle, width, and height, medial/lateral condylar widths, condylar width, medial/lateral posterior tibial slopes and coronal tibial slope. Notch shape index and notch width index were also calculated to adjust for size variations. The predictive accuracy of Risk factors were analyzed using the area under the receiver operating characteristic curve (AUC). The differences of morphological measurements between the two groups were compared. The morphological measurements with statistical significance were included for further multivariate analysis. Risk factors of PCL avulsion fracture were determined by binary logistic regression analysis. The inter- and intra-observer reproducibility of the measurements was determined by the intraclass correlation coefficients (ICC).Results:Except for coronal intercondylar notch height (0.709), medial posterior tibial slope (0.699) and lateral posterior tibial slope (0.705), the other intra-observer ICC ranged from 0.802 to 0.980, which indicated that the parameters had desirable reproducibility. Similarly, except for coronal intercondylar notch height (0.700), the other intra-observer ICC ranged from 0.778 to 0.971. Parameters of patients with PCL avulsion fracture, including the axial notch angle (52.56°±6.56°), coronal notch angle (54.81°±7.29°), axial intercondylar notch width (19.62±2.32 mm), axial notch shape index (0.68±0.08) and medial posterior tibial slope (8.27°±3.53°), were all significantly larger than those of the control group. The binary logistic regression analysis showed that the coronal notch angle ( β=0.102, OR=1.108, P=0.030) and medial posterior tibial slope ( β=0.151, OR=1.163, P=0.046) were the risk factors or PCL avulsion fracture. The AUC for coronal notch angle (0.678) and medial posterior tibial slope (0.631) exhibited good predictive power for the occurrence of PCL avulsion fracture. Conclusion:The larger coronal notch angle and medial posterior tibial slope were positively correlated with the incidence of PCL avulsion fracture.