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Aim of Study: There were many reports published on the relationship between glutathione S-transferase T1 (GSTT1) null/presence gene polymorphism and the risk of lung cancer in these years. In previous, we conducted a meta-analysis to evaluate the relationship between GSTT1 null/presence gene polymorphism and the risk of lung cancer. This study was conducted to update it. Materials and Methods: The association studies were identified from PubMed and Cochrane Library on March 1, 2016. Results: Sixty-three reports were recruited into this meta-analysis for the association of null genotype of GSTT1 with lung cancer susceptibility, consisting of 21,220 patients with lung cancer and 21,496 controls. There was a marked association between GSTT1 null genotype and lung cancer risk in overall populations and in Asians (overall populations: Odds ratio [OR] = 1.17, 95% confidence interval [95% CI]: 1.07–1.28, P = 0.006; Asians: OR = 1.41, 95% CI: 1.23–1.62, P < 0.00001). However, GSTT1 null genotype was not associated with the risk of lung cancer in Caucasians, Brazilian population, and Africans. Conclusion: GSTT1 null genotype is associated with the lung cancer risk in overall populations and in Asians
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BACKGROUND: L-tert-Leucine has been widely used in pharmaceutical, chemical, and other industries as a vital chiral intermediate. Compared with chemical methods, enzymatic methods to produce L-tert-leucine have unparalleled advantages. Previously, we found a novel leucine dehydrogenase from the halophilic thermophile Laceyella sacchari (LsLeuDH) that showed good thermostability and great potential for the synthesis of L-tertleucine in the preliminary study. Hence, we manage to use the LsLeuDH coupling with a formate dehydrogenase from Candida boidinii (CbFDH) in the biosynthesis of L-tert-leucine through reductive amination in the present study. RESULT: The double-plasmid recombinant strain exhibited higher conversion than the single-plasmid recombinant strain when resting cells cultivated in shake flask for 22 h were used. Under the optimized conditions, the double-plasmid recombinant E. coli BL21 (pETDute-FDH-LDH, pACYCDute-FDH) transformed 1 mol·L-1 trimethylpyruvate (TMP) completely into L-tert-leucine with greater than 99.9% ee within 8 h. CONCLUSIONS: The LsLeuDH showed great ability to biosynthesize L-tert-leucine. In addition, it provided a new option for the biosynthesis of L-tert-leucine.
Subject(s)
Leucine Dehydrogenase/metabolism , Bacillales/enzymology , Leucine/biosynthesis , Temperature , Recombinant Proteins , Escherichia coli , Hydrogen-Ion ConcentrationABSTRACT
@#Objective: To explore the mechanism of miR-103 targeting PTEN (gene of phosphate and tension homology deleted on chromsome ten) and activating PI3K/AKT signaling pathway to promote dasatinib (DASA) resistance in lung cancer cells. Methods: DASA-resistant tissues and non-resistant tissues (35 samples for each) from patients treated in Department of Thoracic Surgery, the First Affiliated Hospital of Kunming Medical University from April 2014 to January 2018 were collected for this study. Expression of miR-103 was detected in DASA-resistant tissues and cell lines of lung cancer by quantitative Real-time polymerase chain reaction (qPCR). The effect of miR-103 knock-down on the proliferation, invasion and epithelial mesenchymal transition (EMT) ofA549/DASA cells were measured by CCK-8 assay, Transwell and Wb, respectively. Subsequently, the dual luciferase reporter gene assay was used to verify whether PTEN was a target gene of miR-103. CCK-8, Transwell and Wb assay were further used to investigate the effect of miR103 on malignant biological behaviors of A549/DASA cells via regulating PTEN-PI3K/AKT signaling pathway. Results: miR-103 was highly expressed in DASA-resistant tissues andA549/DASAcells (P<0.01). Knockdown of miR-103 significantly inhibited the proliferation, invasion and EMT ofA549/DASAcells (P<0.05 or P<0.01).Additionally, dual luciferase reporter gene assay confirmed that miR103 directly targeted PTEN and down-regulated its expression (P<0.01). Mechanistically, over-expression of miR-103 targeted and down-regulated PTEN to promote cell viability, invasion and EMT via activating PI3K/AKT pathway (P<0.05 or P<0.01), and further up-regulated the DASA-resistance inA549/DASAcells. Conclusion: miR-103/PTEN/PI3K/AKT signaling pathway plays a certain role in regulating DASA resistance of lung cancer, and knockdown of miR-103 expression may reverse the resistance of A549/DASA cells to DASA.
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Objective@#To understand the active screening of tuberculosis in schools in Guizhou province and analyze the results to provide reference for the prevention and control of tuberculosis in schools in Guizhou province.@*Methods@#In 2017, the initiative screening and entrance examination were carried out among students in Guizhou in 2017, with "the provincial school tuberculosis screening questionnaire" issued by the provincial level.A total of 373 679 students(18.31%) from 290 schools(41.31%)were tested as strong positive by PPD test.@*Results@#Rate of positive PPD skin test showed significant differences according to different types of school(χ2=679.62,P=0.00). the abnormal rate of X-ray chest had statistical significant difference between the students in boarding school or not(χ2=14.07,P=0.00), but had no statistical significant difference between the students in private schools and public schools(χ2=0.28,P=0.59). For the rate of suspicious symptom screening, statistical significant differences were found between the private schools and public schools(χ2=4.79,P=0.03) and boarding schools or not(χ2=23.47,P=0.00). PPD test screening was carried out among 166 691 students, 4 667 were tested as strong positive, 191 cases were found as tuberculosis (4.09%); X-ray chest X-ray screening of 104 024 people, abnormal chest radiograph of 298 people, 200 found that the number of tuberculosis cases, the detection rate was 67.11%. Symptom screening was carried out among 102 964 students, 2 272 had suspicious symptoms, 229 cases were pulmonary tuberculosis patients, the detection rate of 10.08%, three methods of screening for difference was statistically significant(χ2=262.44，P=0.00).@*Conclusion@#The school tuberculosis screening work in Guizhou province needs to be further improved, and the tuberculosis screening for college and boarding high school students should be strengthened to control the outbreak of school tuberculosis.
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Objective To investigate the clinical manifestations,renal pathology and prognosis of antineutrophil cytoplasmic antibody-associated small-vessel vasculitis (AAV) accompanied with renal glomerular IgA deposition.Methods A retrospective analysis was performed at the First Affiliated Hospital of Zhejiang University College of Medicine.Patients diagnosed with AAV associated renal injury by renal biopsy from February 2004 to February 2017 were enrolled.Patients with antiglomerular basement membrane antibody-mediated nephritis,systemic lupus erythematosus nephritis,Henoch Schonlein purpura nephritis,hepatitis B virus associated nephritis and other known etiology were excluded.According to immunofluorescence examination,the patients were divided into IgA deposition group and pauci-immune complex deposition group.The differences in clinical manifestation,pathological features and prognosis were compared between groups.Results A total of 150 AAV cases were included,among which 25 cases were with IgA deposition and 125 cases with pauci-immune complex deposition.The level of serum albumin in IgA deposition group was higher than that in pauci-immune complex deposition group [(35.0±6.2) g/L vs (32.6±5.3) g/L,P=0.049],but the titer of MPO-ANCA was lower [24.8(10.4,71.8) U/ml vs 63.0(21.9,100.0) U/ml,P=0.044] in IgA deposition group.There was no significant difference between two groups in other laboratory indexes and renal pathological findings.The median follow-up time was 15.2 months in IgA deposition group and 8.9 months in pauci immune complex deposition group.During the follow-up there were 8 patients (32.0％) in IgA deposition group and 29 patients (23.2％) in pauci immune complex deposition group on maintaining dialysis;2 patients (8.0％) in IgA deposition group and 7 patients (5.6％) in pauci immune complex deposition group died.There was no significant difference between two groups in patients' outcomes.Conclusions AAV patients with glomerular IgA deposition and AAV patients with typical glomerular immunoglobulin complex deposition are similar as regards clinical appearance and prognosis.
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OBJECTIVE@#To study the effects of Transient receptor potential cation channel, subfamily V, member 1 (TRPV1) combined with lidocaine on status and apoptosis of U87-MG glioma cell line, and explore whether local anesthetic produces neurotoxicity by TRPV1.@*METHODS@#U87-MG cells were divided into control group, gene silencing group, empty vector group and TRPV gene up-regulation group. For cells in each group, flow cytometry was employed to detect the intracellular calcium ion concentration and mitochondrial membrane potential at different time point from cellular perspective. Cell apoptosis of U87-MG was assayed by flow cytometry and MTT from a holistic perspective.@*RESULTS@#Calcium ion concentration increased along with time. The concentration in TRPV1 gene up-regulation group was significantly higher than those in other groups at each time point (P < 0.05). After adding lidocaine, mitochondrial membrane potential in U87-MG significantly increased (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Flow cytometry result and MTT result both showed that cell apoptosis in each group significantly increased after lidocaine was added (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Moreover, apoptosis was more severe along with the increasing concentration of lidocaine (P < 0.05).@*CONCLUSIONS@#In this study, it was proved that lidocaine could dose-dependently induce the increase of intracellular calcium ion concentration, mitochondrial membrane potential and apoptosis in U87-MG glioma cell line. The up-regulation of TRPV1 enhanced cytotoxicity of lidocaine, which revealed the correlations between them. Lidocaine might have increased intracellular calcium ion concentration by activating TRPV1 gene and induced apoptosis of U87-GM glioma cell line by up-regulating mitochondrial membrane potential.
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Objective: To study the effects of Transient receptor potential cation channel, subfamily V, member 1 (TRPV1) combined with lidocaine on status and apoptosis of U87-MG glioma cell line, and explore whether local anesthetic produces neurotoxicity by TRPV1. Methods: U87-MG cells were divided into control group, gene silencing group, empty vector group and TRPV gene up-regulation group. For cells in each group, flow cytometry was employed to detect the intracellular calcium ion concentration and mitochondrial membrane potential at different time point from cellular perspective. Cell apoptosis of U87-MG was assayed by flow cytometry and MTT from a holistic perspective. Results: Calcium ion concentration increased along with time. The concentration in TRPV1 gene up-regulation group was significantly higher than those in other groups at each time point (P < 0.05). After adding lidocaine, mitochondrial membrane potential in U87-MG significantly increased (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Flow cytometry result and MTT result both showed that cell apoptosis in each group significantly increased after lidocaine was added (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Moreover, apoptosis was more severe along with the increasing concentration of lidocaine (P < 0.05). Conclusions: In this study, it was proved that lidocaine could dose-dependently induce the increase of intracellular calcium ion concentration, mitochondrial membrane potential and apoptosis in U87-MG glioma cell line. The up-regulation of TRPV1 enhanced cytotoxicity of lidocaine, which revealed the correlations between them. Lidocaine might have increased intracellular calcium ion concentration by activating TRPV1 gene and induced apoptosis of U87-GM glioma cell line by up-regulating mitochondrial membrane potential.
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Background: Enzymatic decolourization has been recently proposed as a promising and eco-friendly method for treatment of synthetic dye-contaminated wastewaters. However, the processes require large quantities of enzymes, attracting significant attention in developing efficient methods for mass production of multifunctional enzymes. Several methods such as response surface methodology (RSM) and orthogonal experiment have been applied to optimize the parameters in bioprocesses for enzyme production. Results: In the present study, a laccase-like enzyme, phenoxazinone synthase (PHS) originated from Streptomyces antibioticus was recombinantly expressed in Escherichia coli BL21 (DE3). The production of PHS in E. coli BL21 was optimized by response surface methodology based on Box-Behnken design. A full third-order polynomial model was generated by data analysis with Statistica 8.0 in which the optimal conditions for PHS production were calculated to be 1.525 mM CuSO4 and 16.096 hrs induction at temperature of 29.88ºC. The highest PHS production under optimal conditions was calculated to be 4098.51 U/l using the established model. Average PHS production obtained from actual production processes carried out under the calculated optimal conditions was 4052.00 U/l, very close to the value predicted by the model. Crude PHS was subsequently tested in Congo red decolourization which exhibited a low decolourization rate of 27% without mediator. Several mediators were found to improve PHS-catalyzed Congo red decolourization, with the highest rate of 73.89% obtained with 2,2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) as mediator under optimized conditions of 4000 U/l PHS activity, 10 μM ABTS, 100 μM Congo red, and 8 hrs reaction time. Conclusion: Our results indicated that PHS recombinantly produced in E. coli BL21 was a prospective enzyme for decolorizing reactive dye Congo red.
Subject(s)
Oxidoreductases/metabolism , Congo Red/metabolism , Coloring Agents/metabolism , Streptomyces antibioticus/enzymology , Laccase/metabolism , Escherichia coli , WastewaterABSTRACT
The aim of this study is to synthesize the ordered mesoporous silica (OMS) as drug carrier to improve release property of insoluble drug and investigate the dissolution profile of insoluble drug from the porous carrier. The OMS was obtained by using cetyltrimethyl ammonium bromide as the template and resveratrol was selected as the model drug. The resveratrol-loaded OMS (Res-OMS) were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), N2 adsorption-desorption, X-ray diffraction (XRD) and FT-IR spectroscopy. In vitro drug release behavior was also investigated. It was found that the synthesized OMS showed a large surface area, a narrow pore size distribution and an important mesoporosity associated to hexagonally organized channels. Compared with physical mixture and crystalline powder, resveratrol was in amorphous or molecular form after loading into OMS. The release rate ofresveratrol from drug-loaded OMS was significantly increased suggesting the great potential application of OMS for the formulation of poorly soluble drugs.
Subject(s)
Drug Carriers , Drug Compounding , Drug Delivery Systems , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Porosity , Silicon Dioxide , Chemistry , Solubility , Spectroscopy, Fourier Transform Infrared , Stilbenes , Chemistry , X-Ray DiffractionABSTRACT
To study the effect of micro (mi)RNA on cellular proliferation induced by hepatitis B x protein, HBx, in human liver cells and to investigate the underlying molecular mechanism of this cancer-related effect. The human L02 hepatocyte cell line was stably transfected with HBx (L02/HBx) or an HBx mutant (L02/HBx-d382) that induces higher levels of cellular proliferation. The differential miRNA expression profiles were determined by microarray analysis and confirmed by real-time PCR. Two miRNAs, miR-338-3p and miR-551b, that were found to be significantly down-regulated in the L02/HBx-d382 cells were selected for further study and transfected individually into cells using the lipofectamine procedure. The cell survival rate was analyzed by MTT assay, and cell cycles were assessed by flow cytometry. Expressions of cyclinD1, cyclinG1, and E2F1 were assessed by real-time PCR and Western blotting. Compared with the microarray miRNA profile of L02/pcDNA3.0 cells, six miRNAs were up-regulated and five miRNAs were down-regulated in the L02/HBx-d382 cells, while four miRNAs were up-regulated and 12 were down-regulated in the L02/HBx cells. The microarray results were consistent with real-time PCR results. Transfection of miR-338-3p and miR-551b significantly inhibited the cell survival rates (P less than 0.001) and induced G0/G1 phase cycle arrest. According to MTT results: for L02/HBx-d382 cells, compared with lipofectamine or non-transfected (NC) controls, the t value of miR-338-3p was 10.402, 9.133 and the t value of miR-551b was 8.763, 7.403; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 9.105, 8.074 and the t value of miR-551b was 7.673, 7.52. According to flow cytometry results: for L02/HBx-d382 cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 12.173, 11.107 and the t value of miR-551b was 15.364, 13.377; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 15.416, 13.378, and the t value of miR-551b was 13.276, 13.109. The protein levels of cyclinD1, cyclinG1, and E2F1 were significantly reduced by both miR-338-3p and miR-551b ( P less than 0.001). For L02/HBx-d382 cells, compared with lipofectamine or NC controls: E2F1 had t = 11.132, 10.031 and 12.017, 10.973, respectively; cyclinD1 had t = 15.654, 15.013 and 15.447, 14.733, respectively; cyclinG1 had t = 8.017, 7.661 and 7.402, 7.417, respectively. For L02/HBx cells, compared with lipofectamine or NC controls: E2F1 had t = 14.244, 13.331 and 15.022, 14.468, respectively; cyclinD1 had t = 8.695, 8.137 and 7.877, 7.503, respectively; cyclinG1 had t = 7.73, 7.471 and 7.596, 7.41, respectively. In contrast, the mRNA levels for E2F1, cyclinD1, and cylcinG1 showed no significant differences between the miRNA transfected cells and controls. Wild-type HBx and the high proliferation-inducing mutant HBx can influence the miRNA expression profile of L02 cells. HBx down-regulates miR-338-3p and miR-551b in L02 cells, and the high proliferation-inducing mutant has a more robust effect. The mechanism of miR-338-3p- or miR-551b-mediated cell growth inhibition appears to be related to the direct modulation of cyclinD1, cyclinG1, and E2F1.
Subject(s)
Humans , Blotting, Western , Carcinoma, Hepatocellular , Genetics , Metabolism , Pathology , Cell Cycle , Cell Line , Cell Proliferation , Cyclins , Genetics , Metabolism , Gene Expression Regulation, Neoplastic , Genes, Viral , Hepatitis B virus , Genetics , Metabolism , Hepatocytes , Metabolism , Pathology , Liver Neoplasms , Genetics , Metabolism , Pathology , MicroRNAs , Genetics , Metabolism , Mutation , Oligonucleotide Array Sequence Analysis , RNA, Messenger , Genetics , Real-Time Polymerase Chain Reaction , Trans-Activators , Genetics , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To analyze the characteristics of pneumoconiosis cases in Zhejiang province and to provide the evidence for pneumoconiosis control and prevention measures in Zhejiang province.</p><p><b>METHODS</b>The data of new pneumoconiosis cases were from national surveillance system of occupational disease in Zhejiang province during 2006-2009, and were analyzed for distribution, age, exposure duration, pneumoconiosis phases and enterprise types.</p><p><b>RESULTS</b>During 2006-2009, 819 new pneumoconiosis cases (173, 157, 209 and 280 cases, respectively) were reported, 86.9% cases suffered from silicosis. Most of pneumoconiosis cases were distributed in Ningbo, Wenzhou areas and in building materials, machinery, coal, geological and mining, light industries and construction enterprise. The average ages of new pneumoconiosis cases were (47.8 +/- 10.0), (52.5 +/- 13.1), (55.5 +/- 11.2) and (55.9 +/- 12.2) years old, respectively and showed a significant increase trend (P<0.05). The average exposure duration of new pneumoconiosis cases were (12.4 +/- 8.6), (12.9 +/- 9.4), (12.4 +/- 8.6) and (15.7 +/- 10.0) years. The average exposure duration of phase I, phase II, phase III new pneumoconiosis cases were (14.3 +/- 9.87), (12.4 +/- 8.7) and (11.4 +/- 7.1) years, respectively and there were significant differences (P<0.05).</p><p><b>CONCLUSION</b>New pneumoconiosis cases in Zhejiang province are increasing year by year, the main type of pneumoconiosis is silicosis, the distribution of pneumoconiosis cases is associated with the areas and enterprises, and the exposure duration of new pneumoconiosis cases is relatively shorter.</p>
Subject(s)
Adult , Aged , Humans , Middle Aged , China , Epidemiology , Occupational Diseases , Epidemiology , Pneumoconiosis , EpidemiologyABSTRACT
<p><b>OBJECTIVES</b>To study the relationship between quasispecies of hepatitis B virus and the clinical manifestations of their infection, and to find the answer of why different quasispecies of HBV with the same genotype can induce different clinical situations.</p><p><b>METHODS</b>Sixty serum samples, in all of which HBVs of genotype B exist, taken from 32 chronic asymptomatic carriers and 28 severe chronic hepatitis patients, were collected to detect quasispecies of HBV DNA by melt curve approach. Then the relationship between quasispecies of HBV of the same genotype and the clinical situation of their infection was studied by comparing the wave crests of the two sample groups.</p><p><b>RESULTS</b>The data of the 60 serum samples of HBV of genotype B detected by melt curve showed that HBV DNA in severe chronic hepatitis patients had more wave crests than that in chronic asymptomatic carriers (P < 0.05), suggesting that HBV in severe chronic hepatitis patients had more quasispecies than in the chronic asymptomatic carriers.</p><p><b>CONCLUSION</b>The numbers of quasispecies of HBV correlate with the clinical situations of their infection. In the patients infected by HBV of the same genotype, those who have more HBV quasispecies would have more severe clinical manifestations.</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Genotype , Hepatitis B virus , Classification , Genetics , Hepatitis B, Chronic , VirologyABSTRACT
<p><b>OBJECTIVE</b>To establish a cell model of secreted alkaline phosphatase (SEAP) co-controlled by HCV 5'NCR and NS3 serine protease in an effort to develop new antiviral agents.</p><p><b>METHODS</b>The fragments of HCV 5'NCR and NS3/4A-SEAP were amplified by PCR. They were fused into pBluescript SK+ to generate 5'NCR-NS3/4A-SEAP chimeric plasmid. The resulting chimeric gene was subcloned into HindIII/Bsu36 I site of pSEAP2-Control (a SEAP eukaryotic expression plasmid), to generate pNCR-NS3/4A-SEAP, in which the SEAP was fused in-frame to the downstream of NS4A/4B cleavage site. The SEAP activity in the culture media of transiently transfected cells was monitored quantitatively. The regulatory effect of HCV 5'NCR and NS3 serine protease on SEAP expression was measured by treatment of transfected cells with antisense oligodeoxynucleotide (ASODN) against HCV 5'NCR and TPCK, a irreversible serine protease inhibitor.</p><p><b>RESULTS</b>The SEAP activity in the culture media reached 80801+/-4794 RLU, and was significantly inhibited by 5 micromol/L, 10 micromol/L of ASODN (t=4.315, p<0.01; t=6.985, p<0.001) and 100 micromol/L of TPCK (t=6.949, P<0.001).</p><p><b>CONCLUSION</b>A cell model of SEAP co-controlled by HCV 5'NCR and NS3 serine protease has been successfully established. This might promote the screening of anti-viral drugs</p>