Expression of IL-2/IL-15Rβ on memory CD3 + CD8 + CD45RO + T cells in chronic hepatitis B patients undergoing antiviral treatment and its significance / 中华临床感染病杂志

Objective:To investigate the expression of IL-2/IL-15 receptor β subunit (IL-2/IL-15Rβ) on memory CD3 + CD8 + CD45RO + T cells in patients with chronic hepatitis B (CHB) receiving antiviral treatment and its significance. Methods:Sixty-eight patients with chronic active hepatitis B (CAHB) and 47 asymptomatic hepatitis B virus (HBV) carriers attending in the Department of Infectious Diseases, the First Affiliated Hospital of Wannan Medical College from March 2019 to December 2020 were enrolled in the study; and 30 health subjects were also enrolled as healthy control group. Among 60 CAHB patients there were 30 cases with positive HBeAg and 30 cases with negative HBeAg. All CAHB patients received nucleos(t)ide analogue therapy, the HBV-related markers, Alanine aminotransferase (ALT) and the expression of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were determined and compared between HBeAg-positive and negative patients, before and after treatment. Normal distribution measurement data among 3 groups were compared with One-way ANOVA; normal distribution measurement data between 2 groups were compared with paired samples t test; non-normal distribution measurement data between the two groups were compared with Mann-Whitney U test; Pearson’s correlation coefficient was performed for correlation analysis. P<0.05 was considered statistically significant. Results:The proportion of CD8 + CD45RO + T cells on PBMC CD3 + T cells in CAHB group [(8.6±3.7)%] was higher than that of asymptomatic HBV carriers group [(5.7±2.5)%] and healthy control group [(5.5±1.5)%] (all P<0.05). The expression percentage of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group [(6.8±4.7)%] was higher than that of asymptomatic HBV carriers group [(4.7±2.8)%] and healthy control group [(4.3±2.2)%] (all P<0.05). The MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group (243±168) was higher than those of asymptomatic HBV carriers group (160±91) and healthy control group [160±63] (all P<0.05). The expression percentage and MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells were positively correlated with the percentage of CD3 + CD8 + CD45RO + T cells in CAHB patients ( r=0.33 and 0.28, all P<0.05). The proliferation percentage of PBMC CD3 + CD8 + CD45RO + T cells in CAHB group[ (43.7±16.0)%] was higher than that of asymptomatic HBV carriers group [(29.1±9.4)%] and healthy control group [(26.8±9.6)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the proliferation percentage of CD3 + CD8 + CD45RO + T cells was decreased [(11.2±6.3)%] compared with the untreated CAHB group ( P<0.05). The percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ, IL-2 and TNF-α in CAHB group were (13.8±5.4)%, (14.0±4.3)% and (12.3±4.6)% respectively, which were higher than those of asymptomatic HBV carriers [(8.4±2.6)%, (9.4±3.2)% and (6.8±3.3)%] and healthy control group [(6.9±2.7)%, (9.9±3.0)% and (7.7±3.8)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ [(2.4±1.6)%], IL-2 [(4.1±1.9)%] and TNF-α [(4.1±1.8)%] were decreased compared with the untreated CAHB group (all P<0.05). HBeAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were 521.4 (68.9, 1 339.0) COI, 292 (160, 528) U/L, (6.4±3.2)% and (239±136) in 30 HBeAg-positive CAHB patients before treatment, which were higher than those after treatment [3.5(1.5, 17.5)COI、20(14, 31) U/L, (4.1±2.4)% and (134±58)] ( Z=5.337 and 6.403, t=3.229 and 3.892, all P<0.05). HBsAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were (5 310±2 851) COI, (328±207) U/L, (7.1±5.8)% and (252±110) in 30 HBeAg-negative CAHB patients before treatment, which were higher than those after 48 weeks of treatment [(3 811±2 495) COI, (33±14) U/L, (4.6±2.9)% and (154±73)] ( t=2.167, 5.595, 2.116 and 2.383, all P<0.05). Conclusion:The study suggests that up-regulated expression of IL-2/IL-15Rβ is associated with elevated frequency, proliferation and secretion function of memory CD3 + CD8 + CD45RO + T cells in CAHB patients.

Research progress on antiviral therapy in severe hepatic impairment patients and special population with HCV infection / 中华临床感染病杂志

The research and development of direct-acting antiviral agents (DAAs) for hepatitis C achieved significant results in the last decade .DAAs is highly effective in treatment of hepatitic C patients with convenience and safety , and the sustained virological response of DAAs is over 90％, which will greatly contribute to achieving the WHO's goal of eliminating hepatitis C by 2030.The European Association for the Study of the Liver (EASL), American Association for the Study of Liver Diseases (AASLD) and WHO all updated guidelines for diagnosis and treatment of hepatitis C in 2018, which will play a positive role for the management of hepatitis C globally .This article reviews the related guidelines and literature on prevention and treatment of hepatitis C , and summarizes the advantages of antiviral therapy for severe hepatic impairment patients and special population with HCV infection .

CdTe quantum dot inhibits cell survival and induces mitochondrial dysfunction in primary human umbilical vein endothelial cells / 中国药理学与毒理学杂志

OBJECTIVE To elucidate the toxicological properties of CdTe quantum dots (CdTe QD) by investigating their effect on mitophagy in human umbilical vein endothelial cells (HUVECs).METHODS The purity of primarily cultured HUVECs was detected by immunofluorescence.Then,HUVECs were incubated with CdTe QD 0.1-100 mg· L-1 for 24 h.After treatment,the cell viability of HUVECs was detected with MT-T assay.The mitochondrial morphology was observed under a laser scanning confocal microscope after labeling with Mitotracker.The treated HUVECs were also labeled with JC-1 probe,and the mitochondrial membrane potential (MMP) was then examined by flow cytometry.The expression of mitophagy-related proteins including microtubule-associated protein 1 light chain 3 Ⅰ /Ⅱ (LC3 Ⅰ /Ⅱ),moesin-like BCL2-induced protein1(Beclin1),phosphatase and tensin (PTEN) homologinduced putative kinase 1(PINK1) and dynamin-related protein Ⅰ (DRP1) was determined by Westem blotting.RESULTS More than 95％ of the cultured cells expressed vascular endothelial cadherin and herein were vascular endothelial cells.The MTT result showed that the cell survival of HUVECs was significantly decreased after incubation with CdTe QD (0.1-100 mg,L-1) for 24 h (P＜0.05,P＜0.01).CdTe QD also induced extensive fragmentation of the mitochondrial network.The results of JC-1 assay showed that CdTe QD (0.1-100 mg· L-1) caused the disruption of MMP.The percentage of HUVECs with higher MMP was reduced from (91.8±0.6)％ in cell control group to (90.2±1.1)％,(84.4±0.9)％ (P＜0.05) and (78.1 ±1.3)％ (P＜0.01),respectively.The Western blotting data suggested that CdTe QD 10 mg·L-1 significantly increased the expression of autophagy-related protein beclin 1 and the ratio of LC3 Ⅱ/LC3 Ⅰ (P＜0.05,P＜0.01),CdTe QD 1 mg· L-1 also raised the level of mitophagy-related proteins like PINK1 and DRP1.CONCULSION CdTe QD can induce mitochondrial dysfunction as well as mitophagy in HUVECs.

Detection of HIV nucleic acid and subtype analysis in 15 intravenous drug abusers / 中华传染病杂志

Objective To investigate the molecular epidemiology as well as the subtypes of HIV in intravenous drug abusers, which can help pursue the source of infection in Zhejiang province and predict the epidemic strain in the future. Methods Such as ELISA, Western blot, nest PCR, DNA and sequencing technologies were used to analyze HIV subtypes of 15 strains isolated from intravenous drug abusers(IVDU) from Xinjiang autonomous region. Results All of the 15 IVDUs were infected with HIV 1, 2 of which were infected by subtype E and the others were infected by subtype C. Conclusions Subtype C is main HIV subtype infecting IVDUs. The epidemic subtype may have changed in China.

DIFFERENT FACTORS AFFECTING ANTIBODY RESPONSES IN MICE IMMUNIZED BY GENE RECOMBINANT OF HCV STRUCTURE REGION / 免疫学杂志

To seek the optimum experiment methods of animal immunization with HCV gene and to explore the effect on antibody responses in mice immunized by pCD-HCV1 recombinant in different administration, recombinant pCD-HCV1 was constructed by technique of molecular biology and was injected into muscles of Balb/c of mice with different times, routes and dosage of inoculations as well as different treatment. The results showed that the serum antibody level reached 0.183±0.06,0.428±0.05,0.707±0.08 and 0.773±0.07(OD410 value) respectively after recombinant pCD-HCV1(100μg/mouse) were injected into mice once, twice, three times and four times. The antibody level of mice (n＝12) with four times inoculation was the highest; pCD-HCV1 was perfused into stomach orally in mice or were into mice by i.p, s.c and i.m(100μg/mouse, three times) in different routes (n＝6), and the antibody levels were 0.138±0.05, 0.178±0.07, 0.233±0.08 and 0.691±0.05 respectively; after the mice (n＝8) were inoculated with the pCD-HCV1 of different dosage(10μg, 50μg and 100μg) the antibody levels of three groups were 0.11±0.09, 0.33±0.04, and 0.700±0.07, and the results showed a significant difference (P＜0.01); Mice was injected with procaine (100μl, 0.4mg) by i.m or s.c. Then pCD-HCV1 was injected into mice and antibody levels were higher than that of mice immunized directly with recombinant pCD-HCV1 of same dosage. The results may provide a reference data deserved for screening the optimum immunization method of development HCV-DNA-based vaccine in mice model.