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BACKGROUNDA recent outbreak of SARS-CoV-2 infection occurs mainly in China, with rapidly increasing the number of cases (namely COVID-19). Abnormal liver functions are frequently present in these patients, here we aimed to clarify the clinical features of COVID-19-related liver damage to provide some references for the clinical treatment. METHODSIn this retrospective, single-center study, we included all confirmed COVID-19 cases in Shanghai Public Health Clinical Center from January 20 to January 31, 2020. The outcomes were followed up until February 19, 2020. A total of 148 cases were analyzed for clinical features, laboratory parameters (including liver function tests), medications and the length of stay. FINDINGSOf 148 confirmed SARS-CoV-2-infected patients, 49.3% were females and 50.7% were males. The median age was 50.5 years (interquartile range, 36-64). Patients had clinical manifestations of fever (70.1%), cough (45.3%), expectoration (26.7%) at admission. 75 patients (50.7%) showed abnormal liver functions at admission. Patients (n = 75) who had elevated liver function index were more likely to have a moderate-high degree fever (44% vs 27.4%; p = 0.035) and significantly present in male patients (62.67% vs 38.36%; p = 0.005). The numbers of CD4+ and CD8+ T cells were significantly lower in abnormal liver function group than those in normal liver function group. There was no statistical difference in prehospital medications between normal and abnormal liver function groups, while the utilization rate of lopinavir/ritonavir after admission was significantly higher in patients with emerging liver injury than that in patients with normal liver functions. Importantly, the emerging abnormal liver functions after admission caused a prolonged length of stay INTERPRETATIONSARS-CoV-2 may cause the liver function damage and the Lopinavir/ritonavir should be applied carefully for the treatment of COVID-19. FUNDINGShanghai Science and Technology Commission Fund Project and National Science and Technology Major Project
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Hepatitis B virus (HBV) infection is still a major health problem worldwide, but the immunological mechanisms by which HBV causes chronic persistent infection remains controversial. HBV has evolved a series of strategies to interfere TLRs-mediated innate immunity in response to the host immune system and the dysfunction of TLRs is an important cause of persistent virus infection. The innate immune system is activated by pathogens recognition receptors (PRRs) that recognize specific pathogen associated molecular? patterns (PAMPs). Toll-like receptors (TLRs) are important PRRs and play a vital role in mediating innate immune responses to HBV, which is associated with the early clearance of HBV and the subsequent activation of specific immune responses. This review focused on the interplay between HBV and TLRs-mediated innate immune responses as well as research findings about immune therapeutic strategies established based on TLRs.
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ObjectiveTo explore the expression of CD3+ CD8+ human leukocyte antigen (HLA)-A2+T lymphocytes with specificity to the different hepatitis B virus (HBV) peptides in the peripheral blood mononuclear cells (PBMC)from the patients with hepatitis B associated hepatocellular carcinoma (HCC).MethodsThe HLA-A2+ PBMC from four patients with hepatitis B associated HCC were incubated with five HBV/HLA-A2 pentamers respectively,which were HBV sAg (FLLTRILTI),HBV sAg (GLSPTVWLSV),HBV sAg (WLSLLVPFV),HBV core (FLPSDFFPSV),and HBV pol (FLLSLGIHL),as well as anti-CD3-pacific blue and anti-CD8-fluorescein isothiocyanate (FITC).Then,HBV/HLA-A2-CD3-CD8 positive cells were detected by flow cytometry. The monoclonal HBV/HLA-A2-CD3-CD8+ cells were acquired by fluorescenceactivated cell sorter,and cultured and identified by flow cytometry.The anti-HBV specific T lymphocytes were then cultured with HepG2 (HLA-A2+ ) cells and the release of interferon γ (IFN-γ)were determined by enzyme-linked immunosorbent assay (ELISA),Res(a)ltsThe percentage of antiHBV T lymphoeytes with specificity to GLSPTVWLSV in total CD8+ T lymphoeytes from four patients with hepatitis B associated HCC was 1.44%±0.04%,which was higher than those to other four HBV antigen peptides (0.68%±0.08% of FLLTRILTI,1.06%±0.09% of FLPSDFFPSV,0.56% ±0.04% of FLLSLGIHL,and 0.46% ±0.08% of WLSLLVPFV) (t=0.001,P<0.05).The two lines of monoclonal cell with specificity to GLSPTVWLSV both exhibited high level of IFN-γ expression after incubated with hepatic carcinoma cell line HepG2 (HLA-A2+)with HBV GLSPTVWLSV peptide.ConclusionsCD3+ CD8+ HLA-A2+ cells with specificity to the different HBV peptides exist in PBMC of patients with hepatitis B associated HCC.The expression level depends on HBV antigen peptide sequences and genomic sites.
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Objective To investigate the relationship among Helicobacter pylori(H.pylori),CD4 positive cells and CD8 positive cells in gastric mucosa of the AIDS patients with gastritis.Methods Fiftyeight AIDS patients with upper abdominal pain were diagnosed with chronic gastritis through gastroscopy.The gastric biopsies from them were used for H.pylori detection with rapid urease test and Giemsa staining,pathology examination with HE staining,and immunohistochemistry analysis for CD4,CD8 positive cells in Gastric mucosa.And the application of flow cytometry was for the detection of peripheral blood CD4 and CD8 lymphocytes from the patients.Results H.pylori was positive in 26 cases,and negative was in 32 cases.CD8 cell expression in gastric mucosa of the AIDS patients with H.pylori positive was significantly higher than H.pylori negative patients(P<0.05).There is no difference CD4 cell expression in gastric mucosa between the AIDS patients with H.pylori positive and H.pylori negative patients.Moreover,CD8 positive lymphocytes in gastric mucosa of those patients with H.pyloriinfection were significantly stronger than the CD4 positive lymphocytes.However,the peripheral blood CD4 lymphocytes from the patients with H.pylori infection were more than those from H.pylorinegative patients significantly(P<0.05).Conclusion The expression level of CD8 cells in gastric mucosal tissues of AIDS patients with H.pylori infection were higher than those without H.pylori infection.The CD4 lymphocytes from the peripheral blood of the patients with H.pylori infection were more than those without H.pylori negative patients.
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Objective To explore renin expression in cytomegalovirus(CMV) infected juxtaglomerular cells(JG) and its biological significance. Methods JG model cell line As4.1 cells derived from kidney tissue were respectively incubated with murine CMV at multiplicities of infection(MOI) of 10, 0.1 and 0 for 5 d, and the control was mock infection with the same amount of ultraviolet inactive CMV as MOI 10, then the cells were harvested. CMV immediate early gene(IE1) mRNA in the cells was tested by RT-PCR. The renin positive cells and the renin fluorescence granules in the cells were examined by immunofluorescence stain. Whether or not re-nin antigen and CMV antigen were showed in the same cells by FITC and TRITC immunofluorescence. The renin gene expression in the cells was individually detected by real-time RT-PCR and Western blot. Results The cells infected by CMV showed typical cytopathic effect(CPE) and viral plaques in the cell monolayer. CMV IE1 mRNA was found in the viral infected cells by RT-PCR. The mass or ring granules of renin positive fluorescence appeared in the cytoplasm of the CPE cells. The renin positive cells congregated around the viral plaques. Renin positive granules and CMV positive granules showed in the same cells. Renin expression in the CMV infected cells exhibited in a dependent manner of ratio of infectious virus particles to cells. Conclusion CMV infection of the cells derived from kidney tissue induces renin expression related to a new pathogenesis of cardiovascular diseases.
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Objective To study the persistence and replication of hepatitis C virus (HCV) RNA in Epstein Barr virus (EBV) transforming peripheral blood mononuclear cell (PBMC) from patients with he patitis C as well as variation of hypervariable regions (HVR) of the HCV genome. Methods PBMC from one patient with hepatitis C was infected by EBV and then transformed into lymphoblasts capable of being propagated indefinitely. Then, HCV RNA of the cultured cells and supernatants was detected by reverse transcriptase polymerase chain reaction (RT PCR) every month and in situ PCR to identify the location of HCV RNA in the cells. The HVR genome sequences of HCV in the first and the ninth month subculture cells were identified by sequence analysis. Results HCV plus strand RNA could be detected in the cultured cells for as long as 1 year. The HCV plus strand RNA could be identified in supernatants and the minus strand RNA were also observed in the cultured cells intermittently. In situ PCR showed that the blue black positive signals of HCV RNA located mainly in cytoplasmas of EBV transforming B cell but negative in nucleous. The positive signal was not found in negative control cells by in situ RT PCR. HCV HVR genome sequence after half year was found to have two regions of a high degree of variability in 1491 to 1583 nucleotide (384 to 414 amino acid) and 1761~1781 nucleotide (473 to 479 amino acid). But, compared the HCV HVR genome sequence in first month subcultured cells with that in ninth month, there was not significant difference. Conclusion HCV may exist in the cultured cell line for a prolonged period wihtout HVR genome sequence changed.