Objective To detect the expression of metallothionein-3 (MT-3)mRNA in human esophageal squamous cell carcinoma. Methods Five cell lines of human esophageal cancer,TE-1,TE-13,TTN,ECA-109 (cell lines of esophageal squamous cell carcinoma) and OE33 (cell lines of esophageal adenocarcinoma),were used in this study. RT-PCR was employed to detect the expression of MT-3 mRNA. Peripheral blood monouclear cells from normal subjects were served as controls. Results Sequencing of RT-PCR product certified the gene of MT-3 mRNA. It was revealed by gel electrophoresis that there was expression of MT-3 mRNA in each cell line. The relative expression of MT-3 mRNA was 0.230?0.023,0.516?0.020,0.140?0.009,0.176?0.015 and 0.085?0.011 in cell lines of TE-1,TE-13,TTN,ECA-109 and OE33,respectively,significantly lower than that in controls (0.762?0.026) (P

OBJECTIVETo explore the biological effective markers, we investigated DNA strand breaks in peripheral lymphocytes from occupational population with broad ranges of soluble chromate exposure.

METHODSWe conducted a cross-sectional study in the chromate exposed workers employed at a chromate factory in a district of Jinan, Shandong Province. The studied population contained 114 workers from different processes of the chromate plants, in addition, 30 farmers in the countryside about one hundred kilometers away from the factory, without exposure to chromate were matched with the exposed subjects by age, gender and smoking status being identified as a control group. Personal information on age, chromate exposure, medical history (including acute infection and medicine usage), smoking habit and alcohol consumption was obtained by questionnaire. DNA strand breaks in lymphocytes were detected by single-cell gel electrophoresis assay (comet assay) and the DNA damaged degree was evaluated by the score weighted by comet type. The air concentration of chromate was determined by individual sampling for 8 hours per day as shift work and chromium was assayed by atomic absorption spectrometry. The chromium content in the erythrocytes from peripheral blood was determined by graphite furnace atomic absorption spectrometry. The data were analyzed by SPSS10.0 software for statistical significance.

RESULTS(1) The results showed that the score for DNA strand breaks in lymphocytes were 54.52 +/- 23.51 in the exposed group, which was significantly higher than those in the control group (24.70 +/- 11.84) (P < 0.01). (2) The degree of DNA strand breaks in lymphocytes was increased in a dose-dependent manner ranging from 0 microg/m(3) to 106.00 microg/m(3). (3) Correlation analysis showed that there was a significant positive correlation between airborne chromate concentration and the degree of DNA strand break in lymphocytes (P < 0.01). (4) By multiple regression analysis, it was found that the airborne concentrations, chromium contents in red blood cells and smoking habits were factors which might affect the degree of DNA breaks.

CONCLUSIONOur findings suggest that DNA strand break in lymphocytes should be an effective biomarker for occupational chromate-exposed population and be applied in biological monitoring and health risk assessment for occupational chromate-exposed population.

Adult ; Air Pollutants, Occupational ; analysis ; Causality ; Chromates ; analysis ; Chromium ; blood ; Comet Assay ; DNA Damage ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Smoking ; Surveys and Questionnaires

The phyA(m) gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyA(m)-phyCs gene was successfully overexpressed in P. pastoris as an active and extracellular phytase. The yield of total extracellular fusion phytase activity is (25.4+/-0.53) U/ml at the flask scale and (159.1+/-2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 degrees C and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 degrees C to 95 degrees C for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoH(f)), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyA(m).
6-Phytase ; genetics ; isolation & purification ; metabolism ; Amino Acid Sequence ; Fermentation ; Genetic Vectors ; Molecular Sequence Data ; Pichia ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification

Objective To better understand the prevalence and geographic distribution of genotypes/subtypes on HCV and the relationship between HCV genotypes/subtypes and HIV infection disease progression in the HIV-1/HCV co-infected individuals living in high HIV-1 prevalent areas in China. Methods 186 plasma samples were collected from HIV-1 seropositive individuals infected through paid blood donors (PBD), injecting drug users (IDUs) or sexual contact, living in most severely affected provinces, Henan, Yunnan, Xinjiang, Jilin and Liaoning provinces. Samples with HCV viral load >1000 cop/ml were amplified by RT-nested PCR, sequenced and phylogenetically analyzed for genotyping/subtyping of HCV. HIV-1, HCV viral loads and CD4 T lymphocytes were measured for all subjects. Results (1) HCV were identified as 1 a (1.7%), 1 b (39.9%), 2a (17.9%), 3a (10.4%), 3b (15.6%), 6a (1.2%), 6n (6.4%), and a newly unclassified subtype (7.5%). HCV 2a and lb subtypes predominated in PBD in Henan, 3a and 3b in IDUs in Xinjiang and Yunnan, and 6 genotype/subtypes in IDU in Yunnan. (2) There were no significant differences in CD4 T cell counts among the different HCV subtypes. (3) The viral load of HCV RNA in lb subtype was higher than that of non-1b subtype, however, no significant differences in HIV-1 viral loads and CD4 T cell counts were found between Ib and non-1b subtype. Both HIV and HCV viral loads were lower in 2a than non-2a subtype. Conclusion The prevalence of HCV genotype/subtype in HIV-1/FICV co-infected individuals was associated with geographic areas and transmission routes. HCV subtypes had no direct correlation with HIV infection disease progression.

Superior vena cava syndrome(SVCS)is a group of clinical syndromes caused by obstruction of the superior vena cava and its major branches from various causes.Pulmonary artery stenosis(PS)is a complication of lung cancer or mediastinal tumours.SVCS combined with PS due to pulmonary metastases from bladder cancer is extremely rare and has not been reported in the literature.Here we reported an old male patient with pulmonary metastases from bladder cancer presenting with swelling of the head,neck and both upper limbs.SVCS combined with PS was clarified by pulmonary artery computed tomography angiography(CTA)and digital subtraction angiography(DSA).Endovascular stenting was used to treat SVCS.Angiography also showed that PS had not caused pulmonary hypertension and did not need to be treated.The swelling of the patient's head,neck and upper limbs was gradually reduced after the procedure.

Adult ; China ; Databases, Factual ; Female ; Humans ; Male ; Middle Aged ; Pituitary Diseases ; classification ; epidemiology ; Registries ; Software

The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining spermatogenesis. The single-cell RNA sequencing (scRNA-seq) analysis of the testis was performed to identify genes upregulated in spermatogonia. Using scRNA-seq analysis, we identified the spermatogonia upregulated gene origin recognition complex subunit 6 (Orc6), which is involved in DNA replication and cell cycle regulation; its protein expression in the human and mouse testis was detected by western blot and immunofluorescence. To explore the potential function of Orc6 in spermatogonia, the C18-4 cell line was transfected with control or Orc6 siRNA. Subsequently, 5-ethynyl-2-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays, flow cytometry, and western blot were used to evaluate its effects on proliferation and apoptosis. It was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 cells. Bulk RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated (Wnt)/ β-catenin signaling. Western blot revealed that the expression of β-catenin protein and its phosphorylation (Ser675) were significantly decreased when silencing the expression of ORC6. Our findings indicated that Orc6 was upregulated in spermatogonia, whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.