1.Design and implementation of auto-control system for high-throughput and multi-channel DNA synthesis
Xiaohui LIANG ; Chuanwei HUANG ; Jian GUO ; Xiaojiang XIONG ; Xiaoning WANG ; Guiqiang WANG ; Xiaowei FAN
Military Medical Sciences 2015;(8):571-576
Objective To establish an auto-control system for high-throughput and multi-channel DNA synthesis which can simultaneously and quickly synthesize up to 96 different oligonucleotides in a 96-well microtiter format.Methods The PLC and its extended modules is used as the main-control unit, which executes the DNA automatic synthesis process according to the synthesis sequences and steps set by the user,and the manual injecting reagent etc.And the configuration software and VC6.0 were used for programming the man-machine interface sofeware to set synthesis parameters, position calibration,flux calibration data etc, and communicated with PLC.Results The synthesis application of about 150 000 DNA chains has proved that the synthesis cycle time for 96 couplings was 4 min,the average coupling efficiency was 99%across the entire 96-well plate,the monomer reagent usage was reduced by 50 percent,and the synthesis configuration was more flexible.Conclusion A reliable and simple auto-control system is provided for parallel synthesis of 96-channel oligonucleotide chains,which can meet the demands of high-throughput and multi-channel DNA synthesis.
2. Study of the correlation between elevated level of T-bet expression and liver damage in peripheral plasma cells of patients with autoimmune hepatitis
Kangan TAN ; Wanwan SHI ; Liang MIAO ; Xiaoqin DONG ; Wanna YANG ; Hong ZHAO ; Yan WANG ; Guiqiang WANG ; Fengqin HOU ; Yuan HONG
Chinese Journal of Hepatology 2019;27(7):541-546
Objective:
To study the correlation between the level of T-bet expression and liver damage in peripheral plasma cells of patients with autoimmune hepatitis (AIH) in order to provide reference for the study of pathogenesis and development of diseases.
Methods:
The peripheral venous blood and clinical examination data of 29 cases with AIH and 6 healthy volunteers were collected. The percentage of subpopulations of peripheral blood B cells and the proportion of T-bet+ cells in each subgroup were detected by flow cytometry. Plasma cells (CD19+CD10-CD27hiCD38hi), primary B cells (CD19+CD10-CD27-IgD+), transitional B cells (CD19+CD10+), and memory B cells (CD19+CD10-CD27+IgD-) were the included subsets of B cells. Serum immunoglobulin G (IgG) and alanine aminotransferase (ALT) levels, the proportion of B cells in peripheral blood subsets and IgG level, the proportion of T-bet+ cells in each subset and the proportion of T-bet+ plasma cells in each subset in B cells, the proportion of T-bet+ plasma cells and the level of serum ALT were analyzed for correlation analysis. Statistical analysis was performed using two independent sample t-tests and linear regression.
Results:
The serum IgG level of AIH patients with abnormal ALT (19.47 ± 1.039)g/L was significantly higher than that of normal ALT patients (15.5 ± 1.069)g/L, and the difference was statistically significant (
3. Sub-chronic manganese exposure leads to persistent damage of learning and memory ability in rats
Yingnan LÜ ; Qijun WU ; Yuman HUANG ; Pingjing WEN ; Huiyan QIN ; Yumeng FENG ; Jie YANG ; Yunfeng ZOU ; Guiqiang LIANG
China Occupational Medicine 2020;47(01):30-34
OBJECTIVE: To investigate the persistent damage of learning and memory ability after the cessation of sub-chronic manganese(Mn)-exposure in rats. METHODS: Specific pathogen free weaning male SD rats were randomly divided into control group and low-, medium-and high-dose groups based on body weight, with 6 rats in each group. Rats were intraperitoneally injected with Mn chloride(MnCl_2·4 H_2O) at the concentrations of 0, 5, 10, or 20 mg/kg body weight, 5 days per week for 6 weeks and continued to be observed for 12 weeks after the cessation of Mn-exposure. During the experiment, the body mass of the rats was weighed. Learning and memory ability was evaluated by a Morris water-maze task at the 6 th weeks of Mn-exposure(cessation of Mn-exposure of week 0), the 6 th and 12 th week of the cessation of Mn-exposure. The organ coefficients of heart, liver, spleen, kidney and testicles were evaluated after the cessation of Mn-exposure on week 12. RESULTS: The body mass of the high-dose group was lower than that of the other 3 groups(P<0.05) at the 4 th and 6 th week of Mn-exposure and the 2 nd week of the cessation of Mn-exposure. There was no significant difference in body mass between the groups(P>0.05) on the 12 th week of the cessation of Mn-exposure. The escape latency of high-dose group was higher than that of the control group(P<0.05), and the number of platform crossings in the low-, medium-and high-dose groups were fewer than that in the control group(P<0.05) after the cessation of Mn-exposure. The escape latency was shorter and the numbers of platform crossings were higher on the 6 th and 12 th week of the cessation of Mn-exposure(P<0.05) when compared with that of the 6 th week of Mn-exposure rats. There was no statistical significance in the organ coefficients of heart, liver, spleen, kidney and testicles among the 4 groups at the 12 th week of the cessation of Mn-exposure in rats(P>0.05).CONCLUSION: Sub-chronic Mn exposure can impair learning and memory ability of rats, and the damage persists after the cessation of Mn-exposure.
4. Combined anluohuaxianwan and entecavir treatment significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection
Liang MIAO ; Wanna YANG ; Xiaoqin DONG ; Zhanqing ZHANG ; Shibin XIE ; Dazhi ZHANG ; Xuqing ZHANG ; Jun CHENG ; Guo ZHANG ; Weifeng ZHAO ; Qing XIE ; Yingxia LIU ; Anlin MA ; Jun LI ; Jia SHANG ; Lang BAI ; Lihua CAO ; Zhiqiang ZOU ; Jiabin LI ; Fudong LYU ; Hui LIU ; Zhijin WANG ; Mingxiang ZHANG ; Liming CHEN ; Weifeng LIANG ; Hui GAO ; Hui ZHUANG ; Hong ZHAO ; Guiqiang WANG
Chinese Journal of Hepatology 2019;27(7):521-526
Objective:
To explore the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection who received entecavir alone or in combination with anluohuaxianwan for 78 weeks.
Methods:
Patients with chronic HBV infection were randomly treated with entecavir alone or in combination with anluohuaxian for 78 weeks. Ishak fibrosis score was used for blind interpretation of liver biopsy specimens. The improvement in liver fibrosis condition before and after the treatment was compared. Student's t test and non-parametric test (Mann-Whitney U-Test and Kruskal-Wallis test) were used to analyze the measurement data. The categorical variables were analyzed by Chi-square test method and Spearman’s rank correlation coefficient was used to test bivariate associations.
Results:
Liver fibrosis improvement rate after 78 weeks of treatment was 36.53% (80/219) and the progression rate was 23.29% (51/219). The improvement of liver fibrosis was associated to the degree of baseline fibrosis and treatment methods (
5. Clinical effect and safety of pegylated interferon-α-2b injection (Y shape, 40 kD) in treatment of HBeAg-positive chronic hepatitis B patients
Fengqin HOU ; Yalin YIN ; Lingying ZENG ; Jia SHANG ; Guozhong GONG ; Chen PAN ; Mingxiang ZHANG ; Chibiao YIN ; Qing XIE ; Yanzhong PENG ; Shijun CHEN ; Qing MAO ; Yongping CHEN ; Qianguo MAO ; Dazhi ZHANG ; Tao HAN ; Maorong WANG ; Wei ZHAO ; Jiajun LIU ; Ying HAN ; Longfeng ZHAO ; Guanghan LUO ; Jiming ZHANG ; Jie PENG ; Deming TAN ; Zhiwei LI ; Hong TANG ; Hao WANG ; Yuexin ZHANG ; Jun LI ; Lunli ZHANG ; Liang CHEN ; Jidong JIA ; Chengwei CHEN ; Zhen ZHEN ; Baosen LI ; Junqi NIU ; Qinghua MENG ; Hong YUAN ; Yongtao SUN ; Shuchen LI ; Jifang SHENG ; Jun CHENG ; Li SUN ; Guiqiang WANG
Chinese Journal of Hepatology 2017;25(8):589-596
Objective:
To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control.
Methods:
This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (