1.Research advances in new anti-HBV targets and related drugs
Qiang LI ; Qibin ZHUO ; Yuxian HUANG ; Liang CHEN
Chinese Journal of Clinical Infectious Diseases 2016;9(4):379-384
Interferon and nucleotide/nucleoside analogues are currently widely used in the management of chronic hepatitis B virus ( HBV) infections, but are facing problems such as poor sustained virologic response, low HBsAg clearance rate and a high risk of recurrence after drug withdraw.Exploring new target of anti-HBV agent has become a hot topic in recent years.This paper reviews the research progress on new anti-HBV targets and related drugs.
3.Polymorphisms in p.S267F of SLC10A1 gene is associated with clinical outcomes of HBV infection
Qiang LI ; Qibin ZHUO ; Shuye ZHANG ; Yuxian HUANG ; Liang CHEN
Chinese Journal of Clinical Infectious Diseases 2015;(5):425-428
Objective To investigate the association of restriction fragment length polymorphisms (RFLP) in p.S267F of SLC10A1 gene with clinical outcomes of hepatitis B virus (HBV) infection. Methods Clinical data of 1 268 patients with HBV infections admitted in Public Health Clinical Center Affiliated to Fudan University during July 2014 and February 2015 were collected.Polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP) method was used to genotype the p .S267F of SLC10A1 gene in all patients, and the potential association between variants in p .S267F of SLC10A1 gene and the clinical outcomes of HBV infection was analyzed .Results Among 1 268 patients with HBV infections, 1 226 were of genotype CC, and 42 were of genotype CT, so the variation rate in p.S267F was 3.31%(42/1 268).Compared with patients with genotype CC , patients with genotype CT had a higher incidence of acute HBV infections (13.6%vs.28.6%,χ2 =19.819, P<0.05) and a lower incidence of HBV-related liver cirrhosis or hepatocellular carcinoma (13.9% vs.4.8%, χ2 =18.945, P <0.05). Conclusion RFLP in p.S267F of SLC10A1 gene may be associated with chronicity and aggravation of HBV infection, and genotype CT is possibly a protective factor .
4.Magnetic Affinity Immunoassay Based Enzyme-Labeled Phage Displayed Antibody
Xihui MU ; Zhaoyang TONG ; Qibin HUANG ; Bing LIU ; Zhiwei LIU ; Lanqun HAO ; Jinping ZHANG
Chinese Journal of Analytical Chemistry 2014;(6):785-790
A new magnetic affinity immunoassay (MAIA) strategy based on enzyme-labeled phage displayed antibody was developed. The assay consisted of a sandwich format in which immobilized polyclonal antibody (pcAb) on magnetic microparticle was used for capture probe, and enzyme-labeled phage displayed antibody for specific detection probe to increase enzyme amount and enhance detection signal. By the proposed method,β-bungarotoxin (β-BGT) was successfully detected. A linear relationship between absorbance value and the concentration of β-BGT in the range of 0. 016-62. 5 μg / L was obtained. The linear regression equation was Y=0. 641X+1. 355 (R =0. 9925, n = 13, p<0. 0001) with a detection limit of 0. 016 μg / L. In comparison with the traditional ELISA, this method gave a 10-fold better sensitivity in β-BGT detection. This strategy also gave a 4-fold better sensitivity comparing with the MAIA based on enzyme labeled monoclonal antibody (mcAb). Due to low detection limit, acceptable reproducibility and high specificity, this method holds great promise in toxin trace detection.
5.Expression of telomeric-repeat binding factor-1 and telomeric-repeat binding factor-2 mRNA and protein in the peripheral blood mononuclear cells of patients with systemic lupus erythematosus
Mengyun ZHANG ; Yufeng QING ; Shuyue PAN ; Mengjun PU ; Qibin YANG ; Cuipin HUANG
Chinese Journal of Rheumatology 2016;20(9):597-603
Objective To investigate the mRNA and protein expression levels of telomeric-repeat binding factor-1 (TRF1) and TRF2 in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE),and the relations between these gene expression levels and clinical data of SLE patients were explored.Methods According to disease activity,these SLE patients were divided into the active group (40 cases) and the stable group (67 cases).These patients were also grouped as renal damage group (46 cases) and renal damage-free group (61 cases) based on their renal conditions.Healthy individuals (41 cases) were also included as control.Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to study the mRNA expression of TRF1 and TRF2.The protein levels of TRF1 and TRF2 were measured by Western Blot (WB).Independent-Samples t test or one-way analysis of variance (ANOVA) in conjunction with the Least-Significant Difference method (LSD method) wasperformed if the data were in normal distributions;otherwise,the Kruskal-Wallis test was applied.Spearman's correlation analysis was also used for statistical analysis.Results The mRNA and protein expression levels of TRF1 and TRF2 in the PBMCs of the active group (TRF1:0.003 1±0.003 3;TRF2:0.010 5±0.064 8) and renal damage group (TRF1:0.002 3 ±0.002 6;TRF2:0.004 3 ±0.003 3) were significantly increased compared to the stable group (TRF1:0.001 2±0.001 1;TRF2:0.004 2±0.008 6),the renal damage-free group (TRF1:0.001 3±0.001 8;TRF2:0.003 4±0.007 2) and healthy (TRF1:0.001 2±0.003 0;TRF2:0.003 4±0.002 7) individuals respectively (P<0.05).In SLE patients,the expression levels of TRF1 mRNA were correlated with erythrocyte sedimentation rate (r=0.365,P<0.05);the expression levels of TRF2 mRNA were correlated with SLEDAI score (r=0.270,P<0.05),erythrocyte sedimentation rate (r=0.304,P<0.05),creatinine (r=0.258,P<0.05) and 24-hour urinary protein (r=0.344,P<0.05).Conclusion Altered expression of TRF1 and TRF2 might be involved in the pathogenesis of Systemic lupus erythematosus.The positive correlation between TRF2 and SLEDAI score,24-hour urinary protein suggest that TRF2 might be usedas a biomarker for disease activity or renal damage in
6.Expression and purification of mIL-21-hIgGFc fusion protein in 293E cells and its effects on CD8+T cell phenotype
Qibin HUANG ; Mingyue LIU ; Shaoyue FU ; Qiao XING ; Xiaoqi LIU ; Shengdian WANG ; Faping YI
Chinese Journal of Immunology 2014;(7):884-887,892
Objective:To express recombinant protein mIL-21-hIgGFc in 293E cells,and investigate its effect on CD8+T cell.Methods:Total RNA was extracted from the mouse spleen cells ,and then IL-21 gene was amplified by RT-PCR and inserted into expression vector PTT3-hIgGFc.PTT3-mIL-21-hIgGFc were transfected into 293E cells by calcium phosphate method.The supernatants were collected at 48 hours and 72 hours and concentrated by MOLLIPORE Labscale TM TFF system ( 5 kD membrane ).The mIL-21-hIgGFc fusion protein was purified with HiTrap TM Protein G column.The protein was quantified by SDS-PAGE and ELISA.The biological activity of the protein was determined by detecting the change of the phenotypes of CD 8+ T cells treated with the protein.Results: The constructed recombinant plasmid PTT 3-mIL-21-Fc was confirmed by sequencing.PTT3-mIL-21-Fc was transfected into 293E cells,mIL-21-Fc protein in culture supernatant was collected after 48 hours and 72 hours.The protein in cell su-pernatant reached a concentration of 787 ng/ml which was determined by ELISA.The protein was purified by Protein G chromatography column.P1A-specific T cells were treated with mIL-21-hIgGFc, and found that the CD44low CD62Lhi CD8+ population increased compared to the control.Conclusion:We built PTT3-mIL-21-hFc recombinant plasmid, expressed mIL21-hFc fusion protein in 293E cells,and purified by Protein G column.By treating mIL-21-hFc ,the antigen-primed CD8+T cells prefer to differentiate into CD44low CD62Lhi CD8+T cells which had been reported as a memory stem phenotype .This protein may be used to improve the effectness of adoptive T cell cancer therapy.
7.Expression and relationship between matrix metalloproteinase-7 and E-cadherin in gastric cancer
Rongkun DONG ; Du ZHANG ; Jianpeng HUANG ; Mingfu HAN ; Qibin WANG ; Jun FU ; Lin WANG
Cancer Research and Clinic 2011;23(1):25-27
Objective To evaluate the expressions of MMP-7, E-cadherin in gastric carcinoma, and their relationship with tumor cell invasion and metastasis. Methods The MMP-7 mRNA and E-CD mRNA were detected by hybridization in situ in 78 gastric cancer tissues. Then analyze the relationship between the expression of MMP-7, E-CD and clinicopathological features. Results The expressions of MMP-7 and Ecadherin were significantly related with the invasion and metastasis of gastric cancer cell. The E-CD mRNA expression rate in MMP-7 mRNA positive gastric carcinoma tissue was much lower than that in negative carcinoma tissue (9.09 % vs 66.70 %). Conclusion Expression of MMP-7 mRNA and E-CD mRNA is associated with invasion, metastasis. Detection of MMP-7 mRNA and E-CD mRNA in bioptic specimens may be helpful in predicting tumor metastatic and recurrent potential and prognosis for patients with gastric carcinoma.
8.Diagnosis and treatment of hepatocellular carcinoma from a new perspective of miRNAs
Qiang LI ; Qibin ZHUO ; Yuxian HUANG
Journal of Clinical Hepatology 2015;31(6):977-
The hepatocellular carcinoma (HCC) diagnosed by serum biomarkers is usually at late stages, and the five-year survival rate of HCC is extremely low. Therefore, it is necessary to develop an efficient and noninvasive biomarker that can detect HCC at an early stage and to explore a new strategy for the treatment of HCC. Abundant evidence has shown that microRNAs(miRNA)s is involved in the development and progression of HCC, and it can be a sensitive biomarker for the detection of HCC and a new target for the treatment of HCC. The article introduces the application of miRNAs in the diagnosis and treatment of HCC, and believes that the diagnosis and treatment strategy for HCC based on miRNAs has great potential in clinical application, yet remains to be studied.
9.Research advances in therapeutic strategies against covalently closed circular DNA of hepatitis B virus
Qiang LI ; Qibin ZHUO ; Yuxian HUANG
Journal of Clinical Hepatology 2015;31(9):1520-1523
Chronic infection with hepatitis B virus (HBV) often leads to the development of liver cirrhosis and liver cancer, creating immense sociological, clinical, and economic burdens worldwide. Although current anti-HBV drugs can control the disease progression, they often fail to eliminate the virus because of the presence of covalently closed circular DNA (cccDNA) in the hepatocyte nucleus. We review the research advances in therapeutic strategies against cccDNA from the aspects of interfering with cccDNA synthesis, promoting cccDNA degradation, and silencing cccDNA. Targeting cccDNA is a promising approach that may lead to a cure of chronic HBV infection.
10.Clinical features of acute hepatitis B and acute exacerbation of chronic hepatitis B: a comparative study
Qiang LI ; Qibin ZHUO ; Yuxian HUANG
Journal of Clinical Hepatology 2016;32(4):706-710
ObjectiveTo investigate the clinical features of acute hepatitis B (AHB) and acute exacerbation of chronic hepatitis B (CHB) for differential diagnosis. MethodsA retrospective analysis was performed on the clinical data of 96 AHB patients and 124 patients with acute exacerbation of CHB, who were admitted to the Public Health Clinical Center Affiliated to Fudan University from June to December, 2014. Comparison of continuous data between the two groups was made by Mann Whitney U test, while comparison of categorical data was made by chi-square test. ResultsThere were no significant differences in the age of onset and sex between the AHB group and the acute exacerbation of CHB group; the incidence was higher in males than in females. Sexual transmission and iatrogenic transmission were the main routes of transmission for AHB, while mother-to-child transmission was the main route of transmission for acute exacerbation of CHB. The sensitivity and specificity of alanine aminotransferase (ALT) level ≥1072 U/L for diagnosing AHB were 78.6% and 79.2%, respectively. The sensitivity and specificity of S/CO ≥13.6 in the anti-HBc-IgM test for diagnosing AHB were 94.5% and 89.3%, respectively. At week 2 after admission, the AHB group showed significantly greater decreases in levels of HBsAg, HBeAg, and hepatitis B virus (HBV) DNA than the acute exacerbation of CHB group (P<0.05). At week 8 after admission, the AHB group had significantly higher HBsAg clearance rate, anti-HBs seroconversion rate, HBeAg clearance rate, anti-HBe seroconversion rate, and HBV DNA clearance rate than the acute exacerbation of CHB group (P<0.05). ConclusionIt is helpful for making the differential diagnosis between AHB and acute exacerbation of CHB to know the route of transmission, ALT level, anti-HBc-IgM test result (S/CO value), HBV DNA clearance rate, and the seroconversion rates of HBV markers.