Metabolic interventions improve HBV envelope-specific T-cell responses in patients with chronic hepatitis B.

BACKGROUND: Restoration of HBV-specific T cell immunity is a promising approach for the functional cure of chronic Hepatitis B (CHB), necessitating the development of valid assays to boost and monitor HBV-specific T cell responses in patients with CHB. METHODS: We analyzed hepatitis B virus (HBV) core- and envelope (env)-specific T cell responses using in vitro expanded peripheral blood mononuclear cells (PBMCs) from patients with CHB exhibiting different immunological phases, including immune tolerance (IT), immune activation (IA), inactive carrier (IC), and HBeAg-negative hepatitis (ENEG). Additionally, we evaluated the effects of metabolic interventions, including mitochondria-targeted antioxidants (MTA), polyphenolic compounds, and ACAT inhibitors (iACAT), on HBV-specific T-cell functionality. RESULTS: We found that HBV core- and env-specific T cell responses were finely coordinated and more profound in IC and ENEG than in the IT and IA stages. HBV env-specific T cells were more dysfunctional but prone to respond to metabolic interventions using MTA, iACAT, and polyphenolic compounds than HBV core-specific T-cells. The responsiveness of HBV env-specific T cells to metabolic interventions can be predicted by the eosinophil (EO) count and the coefficient of variation of red blood cell distribution width (RDW-CV). CONCLUSION: These findings may provide valuable information for metabolically invigorating HBV-specific T-cells to treat CHB.

Serum cytokine change profile associated with HBsAg loss during combination therapy with PEG-IFN-α in NAs-suppressed chronic hepatitis B patients.

Objective: The aim of this study was to explore the profile of cytokine changes during the combination therapy with pegylated interferon alpha (PEG-IFN-α) and its relationship with HBsAg loss in nucleos(t)ide analogs (NAs)-suppressed chronic hepatitis B patients. Methods: Seventy-six patients with chronic hepatitis B with HBsAg less than 1,500 IU/ml and HBV DNA negative after receiving ≥ 1-year NAs therapy were enrolled. Eighteen patients continued to take NAs monotherapy (the NAs group), and 58 patients received combination therapy with NAs and PEG-IFN-α (the Add-on group). The levels of IFNG, IL1B, IL1RN, IL2, IL4, IL6, IL10, IL12A, IL17A, CCL2, CCL3, CCL5, CXCL8, CXCL10, TNF, and CSF2 in peripheral blood during treatment were detected. Results: At week 48, 0.00% (0/18) in the NAs group and 25.86% (15/58) in the Add-on group achieved HBsAg loss. During 48 weeks of combined treatment, there was a transitory increase in the levels of ALT, IL1RN, IL2, and CCL2. Compared to the NAs group, CXCL8 and CXCL10 in the Add-on group remain higher after rising, yet CCL3 showed a continuously increasing trend. Mild and early increases in IL1B, CCL3, IL17A, IL2, IL4, IL6, and CXCL8 were associated with HBsAg loss or decrease >1 log, while sustained high levels of CCL5 and CXCL10 were associated with poor responses to Add-on therapy at week 48. Conclusions: The serum cytokine change profile is closely related to the response to the combination therapy with PEG-IFN-α and NAs, and may help to reveal the mechanism of functional cure and discover new immunological predictors and new therapeutic targets.

Feasibility of machine learning-based modeling and prediction using multiple centers data to assess intrahepatic cholangiocarcinoma outcomes.

BACKGROUND AND AIMS: Currently, there are still no definitive consensus in the treatment of intrahepatic cholangiocarcinoma (iCCA). This study aimed to build a clinical decision support tool based on machine learning using the Surveillance, Epidemiology, and End Results (SEER) database and the data from the Fifth Medical Center of the PLA General Hospital in China. METHODS: 4,398 eligible patients from the SEER database and 504 eligible patients from the hospital data, who presented with histologically proven iCCA, were enrolled for modeling by cross-validation based on machine learning. All the models were trained using the open-source Python library scikit-survival version 0.16.0. Shapley additive explanations method was used to help clinicians better understand the obtained results. Permutation importance was calculated using library ELI5. RESULTS: All involved treatment modalities could contribute to a better prognosis. Three models were derived and tested using different data sources, with concordance indices of 0.67, 0.69, and 0.73, respectively. The prediction results were consistent with those under actual situations involving randomly selected patients. Model 2, trained using the hospital data, was selected to develop an online tool, due to its advantage in predicting short-term prognosis. CONCLUSION: The prediction model and tool established in this study can be applied to predict the prognosis of iCCA after treatment by inputting the patient's clinical parameters or TNM stages and treatment options, thus contributing to optimal clinical decisions.KEY MESSAGESA prognostic model related to disease staging and treatment mode was conducted using the method of machine learning, based on the big data of multi centers.The online calculator can predict the short-term survival prognosis of intrahepatic cholangiocarcinoma, thus, help to make the best clinical decision.The online calculator built to calculate the mortality risk and overall survival can be easily obtained and applied.

Immunogenicity of inactivated coronavirus disease 2019 vaccines in patients with chronic hepatitis B undergoing antiviral therapy.

Objectives: To investigate the effect and its mechanisms of different antiviral agents on the immunogenicity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines in patients with chronic hepatitis B (CHB). Methods: A total of 125 patients with CHB receiving nucleos(t)ide analogs (NAs) monotherapy or combined with Peg-interferon-alpha (Peg-IFNα) therapy and 29 healthy controls (HCs) were enrolled. Adverse reactions (ADRs) and levels of neutralizing antibody (NAb), immunoglobulin G (IgG), immunoglobulin M (IgM), and peripheral cytokines post-vaccination were analyzed. Results: All ADRs were tolerable in CHB patients. Overall, no significant difference was observed in the antibody levels between patients and HCs after two doses of vaccination. An inverse correlation between NAb, IgG titers and the days after two doses was found in non-IFN group but not in IFN group. Correspondingly, peripheral interferon-γ levels were significantly higher in IFN group than in non-IFN group. After a booster dose, NAb and IgG antibodies were maintained at high levels in NA-treated patients. Conclusion: Peg-interferon-alpha-based therapy may be beneficial for maintaining the immunogenicity of SARS-CoV-2 vaccines in CHB patients, which may be related to the high levels of IFN-γ induced by Peg-IFNα therapy. A booster dose can effectively recall the robust and long-lasting immunogenicity of SARS-CoV-2 vaccines.

A New Prognostic Model Covering All Stages of Intrahepatic Cholangiocarcinoma.

Background and Aims: Intrahepatic cholangiocarcinoma (ICC) is the second most common primary hepatic malignancy that causes a poor survival. We aimed to identify its prognostic factors and to develop a nomogram that will predict survival of ICC patients among all stages. Methods: A total of 442 patients with pathology-proven ICC registered at the Fifth Medical Center of PLA General Hospital between July 2007 and December 2019 were enrolled. Subjects were followed for survival status until June 30, 2020. A prognostic model visualized as a nomogram was constructed in the training cohort using multivariate cox model, and was then validated in the validation cohort. Results: The median age was 55 years. With a median follow-up of 50.4 months, 337 patients died. The median survival was 11.6 months, with 1-, 3- and 5-year survival rates of 48.3%, 22.7% and 16.2%, respectively. Factors associated with overall survival were multiple tumors, lymph node involvement, vascular invasion, distant metastasis, decreased albumin, elevated lactate dehydrogenase (LDH), decreased iron, elevated fibrinogen, elevated CA125 and elevated CA19-9. A nomogram predicting survival of ICC patients at the time of diagnosis achieved a Harrel's c-statistic of 0.758, significantly higher than the 0.582 of the TNM stage alone. Predicted median survivals of those within the low, mid and high-risk subgroups were 35.6, 12.1 and 6.2 months, respectively. Conclusions: A nomogram based on imaging data and serum biomarkers at diagnosis showed good ability to predict survival in patients with all stages of ICC. Further studies are needed to validate the prognostic capability of our new model.

Skewed CD39/CD73/adenosine pathway contributes to B-cell hyperactivation and disease progression in patients with chronic hepatitis B.

BACKGROUND: The mechanisms underlying B-cell hyperactivation in patients with chronic hepatitis B virus (HBV) infection remain largely undefined. The present study assessed the clinical characteristics of the CD39/CD73/adenosine pathway in patients with chronic hepatitis B (CHB). METHODS: We examined CD39 and CD73 expression and adenosine production by B-cells from 202 HBV-infected patients. B-cell-activation phenotypes were assessed by flow cytometry after CpG+CD40 ligand stimulation with or without blockade and activation of the adenosine pathway. RESULTS: CD39 and CD73 expression on circulating B-cells was decreased in CHB patients with high HBV DNA, HBeAg positivity, high HBsAg levels, and active liver inflammation, and was hierarchically restored in complete responders according to HBeAg seroconversion or HBsAg reduction. However, CD39 and CD73 expression on activated memory and tissue-like memory B-cell subsets in complete responders was not increased despite effective antiviral treatments. Furthermore, CD39 and CD73 expression on intra-hepatic B-cells was decreased in inflammatory livers. In vitro, B-cells from CHB patients showed a markedly reduced capacity to generate CD39/CD73-dependent extracellular adenosine and expressed increased levels of activation markers after adenosine-production blockade. Contrastingly, metformin significantly reduced activation-marker expression via regulating AMP-activated protein kinase. CONCLUSIONS: The skewed CD39 and CD73 expression on B-cells was associated with a high viral burden, liver inflammation, and antiviral efficacy in CHB patients, and the skewed CD39/CD73/adenosine pathway contributed to B-cell hyperactivation. Regulation of the CD39/CD73/adenosine pathway using metformin may represent a therapeutic option to reverse HBV-induced immune pathogenesis.

[Effects of light intensity on growth, survival, metabolism and related enzyme activities of Sepia pharaonis.] / å ‰ç §å¼ºåº¦å¯¹è™Žæ–‘ä¹Œè´¼ç”Ÿé•¿ã€å­˜æ´»ã€ä»£è°¢åŠç›¸å ³é ¶æ´»æ€§çš„å½±å“.

An experiment with single-factor design was conducted to investigate the effects of light intensity on growth and survival of cuttlefish (Sepia pharaonis). The specific growth rate, survival rate, oxygen consumption rate, ammonia excretion rate, lactic acid content in muscle, respiratory metabolic enzymes (including hexokinase, pyruvate kinase, and lactate dehydrogenase), supero-xide dismutase, and malondialdehyde in liver were measured in five constant light intensity treatments (10, 30, 50, 70, 90 µmol·m-2·s-1). The main results were as follows: The specific growth rate and survival rate remained steady initially and then decreased gradually with the increases of light intensity. There was no significant difference between groups 10 and 30 µmol·m-2·s-1, but they were significantly higher than those of the other groups. Exposed to light intensities of 10 and 30 µmol·m-2·s-1, the specific growth rates were (8.43±0.22)%·d-1 and (8.47±0.17)%·d-1, and the survival rates were (79.2±5.9)% and (80.0±4.9)%, respectively. Oxygen consumption rates and ammonia excretion rates increased first slowly and then sharply, and reached the maximum value when light intensity was 90 µmol·m-2·s-1, which was significantly higher than those of the other groups. Lactic acid content in muscle firstly decreased and then increased, with the minimum value at 30 µmol·m-2·s-1. The acid content of 10 µmol·m-2·s-1 was significantly lower than those of the other groups except 30 and 50 µmol·m-2·s-1. With the increases of light intensity, the activities of HK and PK in gills remained steady initially and then decreased gradually, and reached the highest level when exposed to 10 and 30 µmol·m-2·s-1, which were significantly higher than those of the other groups. LDH activity in muscle had the lowest level at the light intensity of 10 and 30 µmol·m-2·s-1, which was significantly lower than those of the other groups. SOD activity in liver firstly increased and then decreased, and reached the highest level ((104.93±4.17) U·mg-1 pro) when exposed to 70 µmol·m-2·s-1, which was significantly higher than those of the other groups. MDA content in liver first remained steady and then increased gradually, and reached the highest level ((5.06±0.35) nmol·mg-1 pro) when exposed to 90 µmol·m-2·s-1, which was significantly higher than those of the other groups. In conclusion, the optimum light intensities for growth, survival and metabolism of S. pharaonis were 10 and 30 µmol·m-2·s-1, beyond which S. pharaonis would be under stress. Therefore, sunproof measures should be taken to keep weak light condition in culture practice.

Circulating CXCR3-CCR6-CXCR5+CD4+ T cells are associated with acute allograft rejection in liver transplantation.

Circulating T follicular helper (cTFH) cells have been demonstrated to be involved in B-cell-mediated alloreactive responses in kidney and liver transplantation; however, whether these cells are involved in acute liver allograft rejection after liver transplantation, and which subsets are involved, remains to be clarified. The present study aimed to investigate the profiles of cTFH cells in acute liver allograft rejection, including the CXC motif receptor 3 (CXCR3)+ chemokine receptor 6 (CCR6)- subset, the CXCR3-CCR6- subset, and the CXCR3-CCR6+ subset. Twelve liver transplant patients with acute rejection (AR) and 20 with no acute rejection (NAR) were enrolled in the study. The results showed that the proportion of CXCR3-CCR6-CXCR5+CD4+ T cells was significantly increased and the proportion of CXCR3-CCR6+CXCR5+CD4+ T cells was significantly decreased in patients with AR compared with patients with NAR. In addition, the proportion of CXCR3-CCR6-CXCR5+CD4+ T cells was positively correlated with the proportion of B cells in patients with AR. The level of serum interleukin (IL)-21 was higher in the AR group than in the NAR groups. Furthermore, the proportion of CXCR3-CCR6-CXCR5+CD4+ T cells was positively correlated with alanine amino transferase (ALT), whereas the proportion of CXCR3-CCR6+ CXCR5+CD4+ T cells was negatively correlated with ALT. B cells and TFH cells were detected in follicular-like structures in liver allograft tissues from patients with AR. These results suggest that CXCR3-CCR6-CXCR5+CD4+ T cells may be involved in acute allograft rejection after liver transplantation.

[Effects of light intensity and photoperiod on the embryonic development of Sepia pharaonis.] / å ‰ç §å¼ºåº¦ä¸Žå ‰å‘¨æœŸå¯¹è™Žæ–‘ä¹Œè´¼èƒšèƒŽå‘è‚²çš„å½±å“.

We investigated the effects of lumination on hatching of fertilized eggs of Sepia pharaonis, to reveal the best light conditions for its embryonic development. A single-factor experiment was carried out to examine the effects of different light intensities (10, 30, 50, 70, 90 µmol·m-2·s-1) and different photoperiod L:D (24 h:0 h, 18 h:6 h, 12 h:12 h, 6 h:18 h, 0 h:24 h) on the embryonic development. The results showed that the effects of light intensity on the hatching rate, fractured yolk sac rate, incubation period, mass of newly hatched larvae and mantle length was significant. There was no significant effect on hatching period and survival rate after hatching 7 days. With the increases of light intensity, the hatching rate, incubation period, mass of newly hatched larvae and mantle length first increased and then decreased, while the fractured yolk sac rate gradually increased. The optimum light intensity was 30 µmol·m-2·s-1. Exposed to this light intensity, the hatching rate, fractured yolk sac rate, incubation period, hatching period, mass of newly hatched larvae, mantle length and survival rate after hatching 7 days were (90.0±4.1)%, (7.3±1.5)%, (25.50±0.35) d, (8.10±0.89) d, (0.213±0.011) g, (1.013±0.022) cm, (97.1±4.0)%, respectively. The effects of photoperiod on the hatching rate, incubation period, hatching period were significant, but there was no significant effect on fractured yolk sac rate, mass of newly hatched larvae, mantle length and survival rate after hatching 7 days. With the increases of illumination time, the hatching rate and hatching period first increased and then decreased. The optimum photoperiod was L:D (12 h:12 h). When exposed to this photoperiod environment, the hatching rate, fractured yolk sac rate, incubation period, hatching period, mass of newly hatched larvae, mantle length and survival rate after hatching 7 days were (88.7±1.8)%, (8.7±1.8)%, (25.00±0.50) d, (7.00±3.20) d, (0.209±0.005) g, (0.998±0.026) cm, (96.8±7.1)%, respectively. In conclusion, embryo hatchability of S. pharaonis preferred to low light intensity (30 µmol·m-2·s-1) and normal photoperiod L:D (12 h:12 h). In production practice, sunproof measures should be taken to keep the eggs in weak light condition.

A case of de novo autoimmune hepatitis.

In this study, we reported a case of de novo autoimmune hepatitis. In this case, liver puncture biopsy was carried out and the result showed autoimmune hepatitis. In this report, we described the characteristics of this patient.

[Clinical analysis of liver transplantation for autoimmune hepatitis].

OBJECTIVE: To describe the outcomes and recurrence of autoimmune hepatitis (AIH) after liver transplantation. METHODS: Clinical data of 16 patients with AIH who underwent liver transplantation were analyzed retrospectively. The postoperative cumulative survival rate of the patients was calculated. The postoperative rejections and AIH recurrence were analyzed. The Kaplan-Meier method was used for statistical analysis of survival. RESULTS: All patients were female, with an average age of 52.6 years (range: 41-66 years), and an average MELD score of 21.4. According serological analysis, 15 patients were AIH type 1 and 1 patient was AIH type 2. Three patients died, including 2 of pulmonary infection and 1 of graft dysfunction.The 1-, 2-and 5-year cumulative survival rates were 93.8%, 87.1% and 79.1%, respectively. Five cases (31.3%) of recurrent AIH were diagnosed based on histological evidence. Acute rejection occurred in 6 (37.5%) patients, and de novo HBV infection occurred in 1 (6.3%) patient. CONCLUSION: Liver transplantation is an effective treatment for end-stage AIH. Recurrence and rejection were commonly associated with AIH, but did not negatively impact patient survival.