End-stage liver disease (ESLD) covers the end-stage of acute and chronic liver diseases, mainly involving decompensated cirrhosis, various types of liver failure, and advanced liver cancer. Hepatocyte transplantation has shown promise in treating ESLD, but its clinical application is hampered by the shortage of donor hepatocytes. Cell therapy, an emerging effective treatment for ESLD, faces the same limitation due to scarce hepatocyte availability. Hepatocyte-like cells (HLCs), which are terminally differentiated cells, can be induced from both stem cells and somatic cells. As HLCs exhibit the morphology and function of primary hepatocytes, they offer a promising supplementary source of hepatocytes for cell therapy. First, we provide a background for the differentiation and maturation of primary hepatocytes. Subsequently, based on the current insights into the molecular pathways that regulate hepatocyte differentiation in vivo, we describe a strategy for establishing HLC derived from either stem cells or somatic cells. The key characteristics of these HLCs are also detailed. Furthermore, HLC offers therapeutic potential for liver failure, and HLC-based liver organoids and bioartificial liver systems have demonstrated the ability to provide liver functions, offering an innovative approach to treating various types of ESLD. Despite their promise, challenges such as efficiency in differentiation and functional maturation need to be addressed to improve the clinical application of HLCs. This review discusses these advancements and outlines the therapeutic potential and current challenges of HLC therapy for ESLD.

Japanese encephalitis virus(JEV)belongs to the genus Flavivirus and the family Flavi-viridae,and is classified as a single-stranded,positive-sense RNA virus.The disease known as Japa-nese encephalitis(JE),which results from JEV infection,is a viral zoonosis that is prevalent worldwide and poses a significant public health concern.JEV infection activates a variety of signa-ling pathways,leading to a series of changes that are crucial to the virus's pathogenesis.Among these pathways,Toll-like receptors(TLRs)are particularly significant,and their diverse range and complex signal transduction mechanisms present substantial challenges for the prevention and con-trol of JEV.Currently,there is no specific treatment for JEV.Although some vaccines have been developed to prevent JE,eradicating JEV remains difficult due to its zoonotic transmission cycle and the limited efficacy of the available vaccines.This article reviews the alterations in various TLR signaling pathways induced by JEV infection in the host,aiming to provide insights into the patho-genic mechanisms of JEV and to identify potential new antiviral targets.

Porcine reproductive and respiratory syndrome virus(PRRSV)is the pathogen of porcine reproductive and respiratory syndrome(PRRS),and its infection mainly causes abortion,stillbirth in sows and piglet respiratory infections,which are widely prevalent in the world and seriously jeopardize the development of the world's animal husbandry industry.PRRSV infection of the host is capable of inducing significant immunosuppression,and in recent years,the study of the mecha-nism of PRRSV immunosuppression has become a hot topic,with studies showing that numerous PRRSV proteins are involved in the regulation of host innate immunity and elucidating the mecha-nism by which PRRSV proteins modulate host innate immunity.In this paper,we reviewed the progress of research on the interaction mechanism between PRRSV proteins and host innate im-munity to provide a theoretical basis for a comprehensive understanding of the pathogenesis of PRRSV and the prevention and control of PRRS.

Purpose To investigate the predictive value of placental microflow combined with serum cystatin C(Cys C)for late-onset preeclampsia(LOP).Materials and Methods This was a prospective study including 188 singleton pregnant women who underwent prenatal ultrasound examination and delivered in the First Affiliated Hospital,School of Medicine,Shihezi University from October 2021 to October 2022.The placental microvascular indexes,including vascularization index,flow index and vascularization-flow index(VFI),were detected by three-dimensional power Doppler ultrasound in the second trimester,and the serum Cys C level was detected.The pregnancy outcome of the pregnant women was divided into LOP group and normal group(NLOP group),and the predictive value of placental microflow combined with serum Cys C for LOP was analyzed.Results Placental vascularization index and VFI of the LOP group were lower than those of the NLOP group(29.77±11.97 vs.44.44±9.86,9.23±3.51 vs.15.05±4.38).However,serum Cys C in the LOP group was higher than that in the NLOP group[(1.13±0.17)mg/L vs.(0.84±0.18)mg/L],and the differences were statistically significant(t=-3.616,-4.790,4.682,all P<0.05).The area under the curve of placental VFI combined with serum Cys C for predicting LOP was 0.899,which had the highest predictive value.Conclusion Placental VFI combined with serum Cys C detection can help to improve the prediction efficiency of LOP,and it is better than each single item.

Objective To investigate the correlation between serum sex hormone-binding globulin(SHBG)level and metabolically associated fatty liver disease(MAFLD)in the elderly.Methods A total of 852 patients aged≥60 years who were admitted to the Geriatric Department of Nanjing Drum Tower Hospital were enrolled in this study from June 2015 to October 2023 and divided into MAFLD group(n=426)and non-MAFLD group(n=426).General data,metabolic indexes and serum SHBG were compared between the two groups.The correlation between serum SHBG and various metabolic indexes,the mediating effects of serum SHBG and MAFLD risk,BMI,T2DM and metabolic dysfunction in the association between serum SHBG and elderly MAFLD were analyzed.Results Body mass index(BMI),systolic blood pressure(SBP),diastolic blood pressure(DBP),alanine aminotransferase(ALT),aspartate aminotransferase(AST),glutamyl transpeptidase(γ-GT),serum uric acid(SUA),triglycerides(TG),fasting plasma glucose(FPG),fasting insulin(FIns),glycosylated hemoglobin(HbA1c),insulin resistance index(HOMA-IR),prevalence of T2DM were higher,while high density lipoprotein cholesterol(HDL-C),testosterone(TT)and SHBG were lower in MAFLD group than in non-MAFLD group(P<0.05).Partial correlation analysis showed that SHBG was negatively correlated with BMI,γ-GT,SUA,TG,FIns,HbA1c and HOMA-IR(P<0.05),and positively correlated with LDL-C and HDL-C(P<0.05).Logistic regression analysis showed that after adjusting for confounding factors,a stepwise increase in the OR for MAFLD risk in patients across SHBG 38.00～51.25 nmol/L,27.90～37.90 nmol/L and 6.19～27.85 nmol/L compared with 51.30～147.00 nmol/L.The analysis of mediating effect showed that BMI mediated the effect of SHBG on MAFLD(effect value=-0.0015,P<0.05),and the mediating effect accounted for 25.22%.Conclusions SHBG level is significantly associated with MAFLD in the elderly,and the reduction of SHBG level increases MAFLD risk.

Japanese encephalitis virus(JEV)belongs to the genus Flavivirus and the family Flavi-viridae,and is classified as a single-stranded,positive-sense RNA virus.The disease known as Japa-nese encephalitis(JE),which results from JEV infection,is a viral zoonosis that is prevalent worldwide and poses a significant public health concern.JEV infection activates a variety of signa-ling pathways,leading to a series of changes that are crucial to the virus's pathogenesis.Among these pathways,Toll-like receptors(TLRs)are particularly significant,and their diverse range and complex signal transduction mechanisms present substantial challenges for the prevention and con-trol of JEV.Currently,there is no specific treatment for JEV.Although some vaccines have been developed to prevent JE,eradicating JEV remains difficult due to its zoonotic transmission cycle and the limited efficacy of the available vaccines.This article reviews the alterations in various TLR signaling pathways induced by JEV infection in the host,aiming to provide insights into the patho-genic mechanisms of JEV and to identify potential new antiviral targets.

Porcine reproductive and respiratory syndrome virus(PRRSV)is the pathogen of porcine reproductive and respiratory syndrome(PRRS),and its infection mainly causes abortion,stillbirth in sows and piglet respiratory infections,which are widely prevalent in the world and seriously jeopardize the development of the world's animal husbandry industry.PRRSV infection of the host is capable of inducing significant immunosuppression,and in recent years,the study of the mecha-nism of PRRSV immunosuppression has become a hot topic,with studies showing that numerous PRRSV proteins are involved in the regulation of host innate immunity and elucidating the mecha-nism by which PRRSV proteins modulate host innate immunity.In this paper,we reviewed the progress of research on the interaction mechanism between PRRSV proteins and host innate im-munity to provide a theoretical basis for a comprehensive understanding of the pathogenesis of PRRSV and the prevention and control of PRRS.

Purpose To investigate the predictive value of placental microflow combined with serum cystatin C(Cys C)for late-onset preeclampsia(LOP).Materials and Methods This was a prospective study including 188 singleton pregnant women who underwent prenatal ultrasound examination and delivered in the First Affiliated Hospital,School of Medicine,Shihezi University from October 2021 to October 2022.The placental microvascular indexes,including vascularization index,flow index and vascularization-flow index(VFI),were detected by three-dimensional power Doppler ultrasound in the second trimester,and the serum Cys C level was detected.The pregnancy outcome of the pregnant women was divided into LOP group and normal group(NLOP group),and the predictive value of placental microflow combined with serum Cys C for LOP was analyzed.Results Placental vascularization index and VFI of the LOP group were lower than those of the NLOP group(29.77±11.97 vs.44.44±9.86,9.23±3.51 vs.15.05±4.38).However,serum Cys C in the LOP group was higher than that in the NLOP group[(1.13±0.17)mg/L vs.(0.84±0.18)mg/L],and the differences were statistically significant(t=-3.616,-4.790,4.682,all P<0.05).The area under the curve of placental VFI combined with serum Cys C for predicting LOP was 0.899,which had the highest predictive value.Conclusion Placental VFI combined with serum Cys C detection can help to improve the prediction efficiency of LOP,and it is better than each single item.

Objective:To analyze Hepatitis B virus(HBV)drug resistance mutations in patients with chronic hepatitis B(CHB)infection who have undergone long-term monotherapy with Entecavir(ETV)and those receiving combination therapy with ETV and Lamivudine(LAM), and to explore the related factors affecting HBV drug resistance mutations.Methods:The study retrospectively analyzed patients with CHB, compensated cirrhosis, decompensated cirrhosis, and liver cancer who received long-term nucleotide analogue antiviral therapy at the Fifth Medical Center of PLA General Hospital from August 2012 to August 2019.The patients were divided into an ETV monotherapy group and a combined LAM+ETV therapy group.Chi-square tests, independent sample t-tests, and Wilcoxon rank-sum tests were used to compare the clinical baseline characteristics and HBV drug resistance mutation features between the two therapy groups.A multivariate logistic regression model was used to analyze the factors related to HBV drug resistance mutations. Results:A total of 533 patients were enrolled in this study, 357 in the ETV monotherapy group and 176 in the LAM+ETV group. The ETV monotherapy group had 122 (34.17%) patients with resistance mutations, while the LAM+ETV group had 126 (71.59%).In general, the difference in gene mutation rate between the two therapy groups was statistically significant( χ2=66.337, P<0.001). The median age and alanine aminotransferase levels of patients with drug resistance mutations in the two therapy groups were higher than those in the non-mutation group[( t=-4.743, P<0.001)/( Z=-4.809, P<0.001), ( Z=-2.667, P=0.007)/( Z=-2.001, P=0.045)].Age( OR=1.044, 95% CI:1.023-1.066), compensated cirrhosis( OR=2.163, 95% CI:1.193-3.922), liver cancer( OR=4.017, 95% CI:2.170-7.436) and the treatment regimen( OR=6.075, 95% CI:3.889-9.489) were associated with drug resistance gene mutations( P<0.001).The mutation rates in different stages of chronic liver disease(CHB, cirrhosis, and liver cancer)showed statistically significant( χ2=41.038, P<0.001; χ2=15.894, P<0.001).The overall mutation rates of ETV-related genes in the two therapy groups were 25.49% and 32.39%, respectively.Additionally, 10 mutation sites and 38 variant combinations were identified, containing five common combinations being rtL180M, rtM204V, rtS202G;rtL180M, rtM204V, rtT184A; rtL180M, rtM204V, rtT184L;rtM204I and rtL180M, rtM204V. Conclusion:In CHB patients undergoing long-term therapy, the rate of HBV resistance mutations is higher in those receiving ETV and LAM combination therapy than in those receiving ETV monotherapy.Monitoring older patients and those with cirrhosis or liver cancer is especially important for preventing resistance mutations.

To establish a TaqMan-based multiplex RT-qPCR method for the identification of Japa-nese encephalitis virus(JEV),Porcine reproductive and respiratory syndrome virus(PRRSV),and Classical swine fever virus(CSFV),this study designed and synthesized three pairs of specific primers and probes based on the conserved sequences of JEV E,PRRSV ORF6,and CSFV E2 a-vailable in the NCBI GenBank.By optimizing the reaction system and protocol,a multiplex RT-qPCR method for detecting these three viruses was developed and applied to the detection of clini-cal samples.The results showed that the established TaqMan multiplex RT-qPCR specifically am-plified the gene fragments of JEV,PRRSV,and CSFV,and did not amplify other non-target genes,indicating good specificity of the method.Intra-assay and inter-assay repeatability tests showed that the coefficient of variation(Cv)values were all below 3％,demonstrating that the method has ex-cellent repeatability.Sensitivity tests revealed that the minimum detectable amount for the recom-binant plasmids of the three viruses was 100 copies/pL.Using the established method,a total of 969 samples,including blood,aborted fetuses,semen,and deceased pigs,from 26 pig farms in Guizhou Province were tested.The detection rates were 34.3％(332/969)for JEV,28.3％(274/969)for PRRSV,and 19.8％(192/969)for CSFV.The co-infection rates were 10.1％(98/969)for JEV and PRRSV,12.1％(117/969)for JEV and CSFV,and 14.6％(141/969)for CSFV and PRRSV.Additionally,the triple co-infection rate of JEV,PRRSV,and CSFV was 7.9％(77/969).These results indicate that the TaqMan multiplex RT-qPCR method developed in this study is ef-fective for detecting these three viruses in pig farms,providing technical support for identifying vi-ral causes of reproductive disorders.