Animal models of hepatitis E infection: Advances and challenges.

Hepatitis E virus (HEV) is one of the leading causes of acute viral hepatitis worldwide. Although most of HEV infections are asymptomatic, some patients will develop the symptoms, especially pregnant women, the elderly, and patients with preexisting liver diseases, who often experience anorexia, nausea, vomiting, malaise, abdominal pain, and jaundice. HEV infection may become chronic in immunosuppressed individuals. In addition, HEV infection can also cause several extrahepatic manifestations. HEV exists in a wide range of hosts in nature and can be transmitted across species. Hence, animals susceptible to HEV can be used as models. The establishment of animal models is of great significance for studying HEV transmission, clinical symptoms, extrahepatic manifestations, and therapeutic strategies, which will help us understand the pathogenesis, prevention, and treatment of hepatitis E. This review summarized the animal models of HEV, including pigs, monkeys, rabbits, mice, rats, and other animals. For each animal species, we provided a concise summary of the HEV genotypes that they can be infected with, the cross-species transmission pathways, as well as their role in studying extrahepatic manifestations, prevention, and treatment of HEV infection. The advantages and disadvantages of these animal models were also emphasized. This review offers new perspectives to enhance the current understanding of the research landscape surrounding HEV animal models.

Efficacy and safety of Fuzheng Huayu tablet on persistent advanced liver fibrosis following 2 years entecavir treatment: A single arm clinical objective performance criteria trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Antiviral therapy can alleviate liver fibrosis in chronic hepatitis B, but it has a limited effect on advanced liver fibrosis/cirrhosis. Traditional Chinese medicine (TCM), particularly FuZheng HuaYu (FZHY) tablet, appears to have an antifibrotic effect, but its improving resolution of hepatitis b virus (HBV) -associated advanced fibrosis and experienced anti-viral treatment has not been investigated. AIM OF THE STUDY: To observe the safety and efficacy of adjunctive FZHY on the HBV-associated cirrhosis patients who received 2 years of entecavir but still with advanced fibrosis. METHODS: An open-label, multicentre, single arm trial. 251 patients were included and treated with TCM consisted of FZHY tablets 1.6 g and granules, three times a day in addition to entecavir 0.5 mg daily for an additional 48 weeks. Primary outcome was regression of fibrosis (the proportion of patients with a 1-point decrease in the Ishak liver fibrosis score from baseline to week 48). RESULTS: Fibrosis regression occurred in 94 of 184 patients with paired liver biopsy (51.09%, 95% CI: 43.9～58.0). In 132 compensated cirrhosis patients (Ishak score ≥5), 56.06% (74/132, 95% CI: 47.5～64.2) showed fibrosis regression and reached the goal of 54% (15% more than entecavir mono-therapy). 10 patients occurred adverse reaction, most of them were mild, and all recovered or achieved remission. CONCLUSIONS: The combination therapy of FZHY, TCM granules and ETV could regress the liver fibrosis in the patients with HBV cirrhosis, who experienced 2 years of ETV treatment, and it is safe and well tolerated.

Bacterial infection triggers and complicates acute-on-chronic liver failure in patients with hepatitis B virus-decompensated cirrhosis: A retrospective cohort study.

BACKGROUND: Reports on bacterial infection (BI) in decompensated cirrhosis (DC) is mainly from alcoholic cirrhosis. The role of BI as a trigger or complication of acute-on-chronic liver failure (ACLF) in patients with hepatitis B virus decompensated cirrhosis (HBV-DC) remains to be investigated. AIM: To investigate the impact of BI on the outcomes of the patients with HBV-DC admitted into the hospital with or without ACLF. METHODS: This retrospective study included patients with HBV-DC admitted to two tertiary centers in China. In-hospital overall survival, 90-d transplant-free survival, 5-year post-discharge survival, and cumulative incidence of ACLF were evaluated. Risk factors for death were analyzed considering liver transplantation as a competing event. RESULTS: A total of 1281 hospitalized HBV-DC patients were included; 284 had ACLF at admission. The overall prevalence of BI was 28.1%. The patients with BI had a significantly lower in-hospital survival and transplant-free 90-d survival than those without, in both the patients admitted with and without ACLF. The presence of BI significantly increased the risk of developing ACLF [sub-distribution hazard ratio (sHR) = 2.52, 95%CI: 1.75-3.61, P < 0.001] in the patients without ACLF. In the patients discharged alive, those who had an episode of BI had a significantly lower 5-year transplant-free survival. BI was an independent risk factor for death in the patients admitted without ACLF (sHR = 3.28, 95%CI: 1.93-5.57), while in ACLF admissions, the presence of pneumonia, but not other type of BI, independently increased the risk of death (sHR = 1.87, 95%CI: 1.24-2.82). CONCLUSION: BI triggers ACLF in patients with HBV-DC and significantly impairs short-term survival. HBV-DC patients should be monitored carefully for the development of BI, especially pneumonia, to avoid an adverse outcome.

Myeloid cell-like transcript 2 is related to liver inflammation and the pathogenesis of hepatitis B via the involvement of CD8+T cell activation.

This study analysed the biological significance of TLT-2 on CD8+T cells in hepatitis B patients and provided a theoretical basis for the potential role of TLT-2 as an immune regulator. Flow cytometry sorting, isobaric tags for relative and absolute quantitation and short hairpin RNAs were used to analyse the function of TLT-2 on CD8+T cells in hepatitis B patients. The TLT-2 expression levels in the acute hepatitis B and chronic hepatitis B groups were significantly higher than that in the healthy control group and were positively correlated with ALT and AST. The CD8+TLT-2+T cells exhibited stronger immune function and greater cell proliferation ability and secreted higher levels of cytokines than the CD8+TLT-2-T cells. An analysis of the proteome differences between the TLT-2+CD8+T and TLT-2-CD8+T cells revealed that TLT-2 affected CD8+T cell activation by regulating Granzyme B expression and by further action on the NF-κB signalling pathway. This study first elucidated the mechanism by which TLT-2 influences the activation of CD8+T cells, improved the understanding of the TLT-2 signalling pathway and clarified the role of the TLT-2+CD8+T cell subset in hepatitis B virus infection. The study proposed a novel subset of CD8+T cells that could be useful for understanding the immune function of patients with hepatitis B and further elucidating the pathogenesis of hepatitis B by analysing changes in this subpopulation with the goal of providing a new target for the treatment of hepatitis B.

Clinical Significance of B7-H3 Expression During the Progression of Hepatitis B Virus Infection.

B7-H3, one of the costimulatory members participating in checkpoint pathway, has been shown to be upregulated after hepatitis B virus (HBV) infection. To further explore the clinical significance of dynamic B7-H3 expression during the progression of HBV infection, we systematically investigated the expression pattern of B7-H3 and the correlation of B7-H3 expression with the ratio of T lymphocyte subsets and clinical parameters at different stages in the course of the disease. Flow cytometry and enzyme-linked immunosorbent assay data showed that soluble form of B7-H3 (sB7-H3) was positively correlated with the frequency of Treg cells in acute hepatitis B (AHB), chronic hepatitis B (CHB), and hepatocellular carcinoma patients with HBV infection (HBV-HCC). Membrane form of B7-H3 (mB7-H3) expressed on Treg cells and monocytes was positively correlated with the frequency of Treg cells in CHB. SB7-H3 had relationship with mB7-H3 expressed on Treg cells and monocytes at different stages during HBV infection, except for HBV-HCC. MB7-H3 expressed on Treg cells was positively correlated with that on monocytes in AHB, CHB, HBV-liver cirrhosis, and HBV-HCC. The B7-H3 expression was positively correlated with aspartate aminotransferase and alanine aminotransferase levels in CHB and sB7-H3 level was higher in late tumor/node/metastasis (TNM) stage in HCC. Higher mB7-H3 expression was associated with greater tumor size, later TNM stage, and worse prognosis in HBV-HCC indicated by immunohistochemistry. Taken together, these results suggested that B7-H3 might contribute to the progression of HBV infection by triggering inhibitory signals in effector T cells and it was closely associated with the progression and poor prognosis during HBV infection. B7-H3 could be utilized as a potential clinical indicator and a potential target for therapeutic strategies against HBV infection.

The Expression of B7-H3 in Circulating CD4+CD25high T Cells, Circulating CD14+ Monocytes and Plasma during Hepatitis B Virus Infection Progression.

BACKGROUND: B7-H3 is frequently upregulated in response to autoantigens and pathogens during host T cell immune responses. However, the role of B7-H3 which express in CD14+ monocytes and CD4+CD25high T cells had not been investigated. METHODS: Cytometry and ELISA were used in this study. RESULTS: The study showed that B7-H3 expression in CD14+ monocytes, CD4+CD25high T cells, and plasma was significantly increased in AHB, CHB, HBV-LC, and HBV-HCC group. In CHB group, the expression of B7-H3 was positively correlated with the ALT and AST levels. CONCLUSIONS: B7-H3 expression in peripheral CD14+ monocytes, CD4+CD25high T cells, and plasma changed with HBV infection progression and had a significant correlation with liver function in CHB. B7-H3 expression could be utilized as a potential clinical indicator to determine the extent of liver injury.

Interferon alpha on expression of hTERT mRNA in peripheral blood mononuclear cells of patients with chronic hepatitis B.

Cell division is closely related to telomerase activity (hTERT mRNA). Lower expression of lymphocitic hTERT mRNA may easily cause cell aging, which is not beneficial to maintaining a durable lymphocyte division. To date, there is no study to investigate IFNα therapy on hTERT mRNA expression in PBMCs of patients with chronic hepatitis B (CHB). We quantitatively detected hTERT mRNA from study subjects and made each hTERT mRNA normalized (NhTERT mRNA). Mean NhTERT mRNA level was lower in either CHB group, but it significantly increased in IFNα-treated group compared with CHB control group, and a longer duration of IFNα therapy could increase the level. Moreover, the mean NhTERT mRNA in subgroup with HBeAg loss was significantly higher than that in subgroup without. NhTERT mRNA was markedly correlated with CD3(+) T lymphocyte count and CD4(+)/CD8(+) ratio. The results showed that IFNα therapy could upregulate the expression of hTERT mRNA in PBMCs.

[Effects of Wulongdan on learning and memory abilities of rats with chronic cerebral ischemia].

OBJECTIVE: To evaluate the effect of Wulongdan, a tradition Chinese medicinal preparation, on the learning and memory abilities of rats with chronic cerebral ischemia and explore the mechanisms. METHODS: Male rats with chronic cerebral ischemia induced by permanent ligation of the bilateral carotid arteries were randomized into sham-operated group, chronic cerebral ischemia (model) group, and high-, middle-, and low-dose Wulongdan groups and Yinxingye group. The corresponding treatments were administered in the rats 24 h after the operation once daily for 8 consecutive weeks. Morris water maze and step-through tests were performed after 7 weeks of drug administration. The brain tissues were then taken to observe the morphological changes in the hippocampal neurons with Nissl staining by transmission electron microscopy. RESULTS: Compared with the model group, the latency of finding the platform in Morris water maze test was significantly shortened (P<0.05 or 0.01), and that in step-through test significantly prolonged (P<0.05 or 0.01) in high-, middle-, and low-dose Wulongdan groups and Yinxingye group. In the model group, Nissl staining of the hippocampal CA1 region visualized obvious pathological changes in the neurons, showing a significant difference from the sham-operated and high-dose Wulongdan groups. CONCLUSION: Wulongdan can enhance the learning and memory abilities of rats with chronic cerebral ischemia possibly through a mechanism in relation to neuronal protection in the hippocampus CA1 region.

[CD4+CD25+high regulatory T cells in patients with autoimmune hepatitis].

OBJECTIVE: To investigate the role of CD4+CD25+high regulatory T cells in the pathogenesis of autoimmune hepatitis. METHODS: CD4+CD25+ high regulatory T cells and CD4+ T cells were measured by using flow cytometry in 16 patients with autoimmune hepatitis, 22 patients with chronic hepatitis B and 20 healthy blood donors. Foxp3 protein was detected by immunohistochemical assay in liver tissues from the patients with autoimmune hepatitis or chronic hepatitis B. RESULTS: The percentage of CD4+CD25+high/CD4+ in patients with autoimmune hepatitis was significantly lower than that in healthy controls and patients with chronic hepatitis B. Meanwhile, the percentage of CD4+CD25+high/CD4+ highly increased in patients with chronic hepatitis B, compared with healthy controls; Foxp3 positive cells were mostly located in the hepatic lobular perisinusoidal spaces and the portal tract, and there was a significant difference in the quantity of Foxp3 positive cells between patients with autoimmune hepatitis and chronic hepatitis B. CONCLUSION: Patients with autoimmune hepatitis harbor a decreased percentage of CD4+CD25+ high regulatory T cells, which may be associated with development of autoimmunity.

[The relationship between the efficacy of interferon-alpha treatment and the variation of perforin protein expression in peripheral blood mononuclear cells in chronic hepatitis B patients].

OBJECTIVE: To study the relationship between the efficacy of interferon-alpha and the variation of perforin protein expression in the peripheral blood mononuclear cells in 35 patients with chronic hepatitis B (CHB). METHODS: The perforin protein in the peripheral blood mononuclear cells was detected by immunocytochemistry technique. RESULTS: The level of the perforin protein expression in the peripheral blood mononuclear cells was significantly higher in the post-treatment group with interferon-alpha than in the pre-treatment group. At the end of the treatment with interferon-alpha, there were 12 cases of complete responders, 14 cases of partial responders, and 9 cases of non-responders. After interferon-alpha treatment, the mean level of the perforin protein expression in the peripheral blood mononuclear cells was 12.1%, 6.9% and 3.9% respectively, and there existed significant differences among the three groups. Moreover, before treatment, the level of the perforin protein expression in the complete responder group was significantly higher compared to the partial responder group or the non-responder group. CONCLUSION: Treatment with interferon-alpha can increase the perforin protein expression in the peripheral blood mononuclear cells in patients with CHB. The variation of perforin protein expression in the peripheral blood mononuclear cells may be closely related to the efficacy of interferon-alpha treatment against hepatitis B virus.

[Effects of Buyanghuanwu decoction on nerve proliferation in rats with sequelae of ischemic stroke].

OBJECTIVE: To examine the effect of Buyanghuanwu decoction (BYHWD) in inducing nerve proliferation in rats with sequelae of ischemic stroke. METHODS: A rat model of ischemic stroke sequelae was established by means of craniectomy in which the right common carotid artery was ligated with 4-0 silk thread followed by cauterization of the right middle cerebral artery. Programmed electric shock was administered 24 h after the onset of ischemic stroke for 2 h daily for 20 consecutive days. The rats in sham operation group were not subjected to ligation of the right common carotid artery or right middle cerebral artery occlusion. The rats in the treatment groups were given oral BYHWD for 15 consecutive days. All the rats received repeated intraperitoneal injections of the cell proliferation-specific marker 5-bromodeoxyuridine (BrdU), and the intake of BrdU in the cerebral tissues was determined by immunohistochemistry. RESULTS: The number of BrdU-immunoreactive cells in the cerebral tissues of BYHWD-treated rats was significantly greater than that in the untreated model group. CONCLUSION: BYHWD can promote nerve proliferation in rats with ischemic stroke sequelae.

[Effect of interferon-alpha therapy on the capacity of antigen presenting of peripheral blood dendritic cells from patients with chronic hepatitis B].

OBJECTIVE: To study the effect of interferon -alpha therapy on the capacity of antigen presenting of peripheral blood dendritic cells from patients with chronic hepatitis B (CHB). METHODS: The peripheral blood samples were obtained from 23 patients who were given interferon-alpha therapy just before treatment and after treatment for 4 months, respectively. The peripheral blood mononuclear cells (PBMC) were isolated and cultured, and recombinant human IL-4 and GM-CSF were added to the cultures. After cultured for 7 days, dendritic cells (DC) were harvested and then incubated with HBsAg for 3 hours, then mixed with autogenous PBMC and cocultured for additional 72 hours. Before ending the culiration, 7.4 x 10(4) Bq (3)H-TDR was added to the culture for 12 hours, and then all cells were collected and detected for cpm values. Eight healthy individuals were used as controls. RESULTS: After treatment for 4 months with interferon-alpha, the proliferating level of DC markedly increased in posttreatent group when compared with that in the pretreatment group and the total number of DC proliferation averagely increased 2.8 times in the same culture condition. The capacity of antigen presenting of DC in the pretreatment group markedly decreased compared with that either in posttreatment group or in healthy group, respectively (P < 0.001), and there was no significant difference between the posttreatment group and the healthy group (P > 0.05). Both before and after treatment the capacities of DC antigen presenting in interferon-alpha complete responder group were significantly stronger than those in the nonresponder group (P < 0.01 and P < 0.001). There was no significant difference between the partial responder group and nonresponder one (P > 0.05). CONCLUSION: The results indicate the capacity of antigen presenting of peripheral blood DC from CHB patients is dysfunctional. Interferon-alpha therapy may markedly improve the capacity. The potential of antigen presenting of DC in CHB patients may be closely correlated with the response to interferon-alpha therapy.