Expression of HBcAg in hepatocytes and its association with the efficacy of antiviral therapy / 临床肝胆病杂志

ObjectiveTo investigate the effect of the expression of HBcAg in hepatocytes on the serum level of HBcAb and seroconversion of HBeAg after antiviral therapy with nucleos(t)ide analogues (NUCs). MethodsSerum samples and liver tissue paraffin sections were collected from 101 chronic hepatitis B (CHB) patients who received antiviral therapy with NUCs in Nanfang Hospital and Panyu Central Hospital from January 2015 to June 2018. ELISA was used to measure the serum level of HBcAb, and immunohistochemistry was used to measure the expression of HBcAg in the liver. The GEO database (GSE96851) was analyzed to obtain differentially expressed genes in the liver of patients with HBcAg-positive hepatitis. The two-independent-samples t test was used for comparison of continuous data between two groups; the multiple-independent-samples nonparametric Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and Dunnett method was used for further comparisons; the chi-square test was used for comparison of categorical data between groups. ResultsThe expression pattern of HBcAg in hepatocytes was classified as absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression, and according to expression level, HBcAg expression was classified as grades Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression. HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 16.67%, 22.73%, and 24.24%, respectively, in the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression (χ2=4753, P=0.037), and HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 13.04%, 27.59%, and 26.67%, respectively, in the patients with grade Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression (χ2=6.580, P=0.016). There were significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression of HBcAg (HBcAb-IgM: H=9.760, P=0.021; total HBcAb: H=21.46, P＜0.001), and there were also significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with grade Ⅰ, Ⅱ, Ⅲ, and IV expression of HBcAg (HBcAb-IgM: H=18.80, P＜0.001; total HBcAb: H=26.03, P＜0.001). The analysis of differentially expressed genes in the liver showed that the expression of antibody-related genes was upregulated in the liver of patients with HBcAg-positive acute liver failure. ConclusionThe expression pattern and level of HBcAg in the cytoplasm of hepatocytes are associated with serum HBcAb, and the measurement of HBcAg may help to predict the efficacy of antiviral therapy with NUCs.

Construction of full-length human bladder cancer-specific antibody libraries based on mammalian display technology / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.</p><p><b>METHODS</b>The total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.</p><p><b>RESULTS</b>The libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).</p><p><b>CONCLUSION</b>We have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.</p>

Study on Correlativity of TCM Syndrome Types with Sexual Hormones in Patient of Hyperplasia of Mammary Glands / 中医杂志

Objective:To explore relation of TCM syndrome types in patient with sexual hormones of hyperplasia of mammary glands,so as to establish objective indexes of TCM syndrome types of hyperplasia of mammary glands.Methods:112 cases of hyperplasia of mammary glands were divided into 3 types:stagnation of liver-Qi,intermingled phlegm and blood stasis,disharmany of thoroughfare vessel and conception vessel.Level of estradiol(E_2),progestin(PT),testosterone(TT),prolactin(PRL),follicle- stimulating hormone(FSH)and luteotropic hormone(LH)during ovulatory period in the patients were determined.The relation of hormone level with TCM syndrome types was analyzed.Results:Level of LH and E_2 decreased,PRL and PT increased and level of TT and FSH did not significantly change during ovulatory period in the patient of hyperplasia of mammary glands.LH and E_2 level in the patients with stagnation of liver-Qi or intermingled phlegm and blood stasis was significantly lower than normal values,and the level of PRL and PT was significantly higher than that of normal values.The level of LH and E_2 was also lower than the normal value in the patient with disharmany of thoroughfare vessel and conception vessel,but the level of FSH was significantly higher than normal value.Conclusion:The patient of hyperplasia of mammary glands during ovulatory period manifests endocrine dysfunction,and the patients with different TCM syndrome are different in secretion of sexual hormones; Changes of sexual hormones can be used as objective indexes of TCM syndrome types and basis of clinical medicinal application.