Anisodine hydrobromide alleviates oxidative stress caused by hypoxia/reoxygenation in human cerebral microvascular endothelial cells predominantly via inhibition of muscarinic acetylcholine receptor 4

Background:Anisodine hydrobromide(AT3),an anti-cholinergic agent,could be delivered to the brain across the blood-brain barrier and has been used clinically for the treatment of cerebral ischemia/reperfusion injury.Endothelial dysfunction can be caused by hypoxia/reoxygenation(H/R)via oxidative stress and metabolic alterations.The present study investigated whether AT3 regulates the production of nitric oxide(NO)and reactive oxygen species(ROS),and the HIF-1αpathway via regulation of muscarinic acetylcholine receptors(mAChRs)in brain microvascular endothelial cells after H/R exposure.Methods:Under H/R conditions,hCMEC/D3 cerebral microvascular endothelial cells were treated with AT3.Specific inhibitors of M2-and M4-mAChRs were used to explore the mechanism by which AT3 influences oxidative stress in endothelial cells.Then,mAChRs expression was detected by western blotting and NO production was detected by Greiss reaction.The intracellular ROS level was measured using DCFH-DA probes.The expression of hypoxia-inducible transcription factor 1α(HIF-1α)was also detected.Results:While H/R induced the expression of M2-and M4-mAChRs,AT3 suppressed the H/R-upregulated M2-and M4-mAChRs.H/R also induced the production of NO,ROS,and apoptosis.AT3 and M4-mAChR inhibitors inhibited the H/R-induced production of NO and ROS and apoptosis.HIF-1αwas induced by H/R,but was suppressed by AT3.Conclusion:Thus,the in vitro evidence shows that AT3 protects against H/R injury in cerebral microvascular endothelial cells via inhibition of HIF-1α,NO and ROS,predominantly through the downregulation of M4-mAChR.The findings offer novel understandings regarding AT3-mediated attenuation of endothelial cell apoptosis and cerebral ischemia/reperfusion injury.

A novel nine gene signature integrates stemness characteristics associated with prognosis in hepatocellular carcinoma

Cancer stem cells(CSCs)are heterogeneous with self-renewal and differentiation ability.The mRNA expression-based stemness index(mRNAsi)described the similarity between tumor cells and CSCs,which is positively associated with the poor prognosis of cancer patients.However,the key prognostic genes related to mRNAsi in hepatocellular carcinoma(HCC)remains unclear.A 9-gene signature related to mRNAsi and HCC prognosis including PSMG3,SNRPD1,DTYMK,PIGU,NME1,TXNL4A,IPO4,PES1,and REXO4 was obtained.High expression of this signature indicates poor prognosis of HCC.PIGU was an independent prognostic factor of HCC,which was significantly associated with progression of HCC.Among them,DTYMK and NME1 enriched in pyrimidine metabolism,SNRPD1 and TXNL4A enriched in spliceosome and PIGU enriched in glycosyl phosphatidylinositol(GPI)-anchor biosynthesis pathways.High levels of IPO4,NME1,PES1,PIGU and SNRPD1 were closely associated with metastasis of HCC,and low levels of IPO4,PIGU and REOX4 were significantly associated with sorafenib resistance of HCC.High expression of the 9-gene signature was negatively correlated with the stromal cell infiltration,and positively correlated with specific immune subtypes-related to angiogenesis,M1/M2 macrophage polarization,and M2 response.The 9-gene signature was negatively correlated with the stroma,and SNRPD1 and TXNL4 were positively correlated with immune infiltrate.NME1 was negatively correlated with tumor purity.Therefore,a 9-gene signature related to mRNAsi and poor prognosis in HCC were identified,which can be used as biomarkers for the diagnosis of HCC and functional mechanism exploration of CSCs in HCC.These genes such as IPO4 and PIGU might drive the transition of tumor cells into CSCs which possibly controls the balance between metastasis and drug resistance in HCC.The challenge on balance between metastasis and drug resistance for tumor therapy was firstly reported by the present study.

Glutamine synthetase-negative hepatocellular carcinoma has better prognosis and response to sorafenib treatment after hepatectomy

Background:Glutamine synthetase(GS)and arginase 1(Arg1)are widely used pathological markers that discriminate hepatocellular carcinoma(HCC)from intrahepatic cholangiocarcinoma;however,their clinical significance in HCC remains unclear.Methods:We retrospectively analyzed 431 HCC patients:251 received hepatectomy alone,and the other 180 received sorafenib as adjuvant treatment after hepatectomy.Expression of GS and Arg1 in tumor specimens was evaluated using immunostaining.mRNA sequencing and immunostaining to detect progenitor markers(cytokeratin 19[CK19]and epithelial cell adhesion molecule[EpCAM])and mutant TP53 were also conducted.Results:Up to 72.4%(312/431)of HCC tumors were GS positive(GS+).Of the patients receiving hepatectomy alone,GS negative(GS-)patients had significantly better overall survival(OS)and recurrence-free survival(RFS)than GS+patients;negative expression of Arg1,which is exclusively expressed in GS-hepatocytes in the healthy liver,had a negative effect on prognosis.Of the patients with a high risk of recurrence who received additional sorafenib treatment,GS-patients tended to have better RFS than GS+patients,regardless of the expression status of Arg1.GS+HCC tumors exhibit many features of the established proliferation molecular stratification subtype,including poor differentiation,high alpha-fetoprotein levels,increased progenitor tumor cells,TP53 mutation,and upregulation of multiple tumor-related signaling pathways.Conclusions:GS-HCC patients have a better prognosis and are more likely to benefit from sorafenib treatment after hepatectomy.Immunostaining of GS may provide a simple and applicable approach for HCC molecular stratification to predict prognosis and guide targeted therapy.

Effects of viral load on tenofovir vs.entecavir efficacy in recurrence prevention of hepatitis B virus-related hepatocellular carcinoma

Serum hepatitis B virus deoxyribonucleic acid(HBVDNA)load is a critical index used to reflect the degree of viral replication in hepatocytes and to evaluate host hepatitis viral status.The risk of hepatocellular carcinoma(HCC)incidence or recurrence varies depending on whether the HBV-DNA load is high or low.[1]Consistent with previous studies and expert opinion,a HBV-DNA level>2000 IU/mL was defined as a high HBV-DNA load in the current study.

Tenofovir vs.entecavir on recurrence of hepatitis B virus-related hepatocellular carcinoma beyond Milan criteria after hepatectomy

Background:Hepatectomy for hepatocellular carcinoma(HCC)beyond the Milan criteria is shown to be beneficial.However,a high rate of post-operative HCC recurrence hinders the long-term survival of the patients.This study aimed to investigate and compare the impacts of tenofovir(TDF)and entecavir(ETV)on the recurrence of hepatitis B viral(HBV)-related HCC beyond the Milan criteria.Methods:Data pertaining to 1532 patients who underwent hepatectomy and received antiviral therapy between January 2014 and January 2019 were collected from five centers.Recurrence-free survival(RFS)analysis was performed using the Kaplan-Meier method.Cox proportional hazards regression analysis was performed to determine prognostic factors for HCC recurrence.Results:The analysis incorporates 595 HBV-related HCC patients.The overall 5-year RFS was 21.3%.Among them,533 and 62 patients received ETV and TDF treatment,respectively.The 1-,3-,and 5-year RFS rates were 46.3%,27.4%,and 19.6%,respectively,in the ETV group compared with 65.1%,41.8%,and 37.2%,respectively,in the TDF group(P<0.001).Multivariate analysis showed that TDF treatment(hazard ratio[HR]:0.604,P=0.005),cirrhosis(HR:1.557,P=0.004),tumor size(HR:1.037,P=0.008),microvascular invasion(MVI)(HR:1.403,P=0.002),portal vein tumor thrombus(PVTT)(HR:1.358,P=0.012),capsular invasion(HR:1.228,P=0.040),and creatinine levels(CREA)(HR:0.993,P=0.031)were statistically significant prognostic factors associated with RFS.Conclusions:Patients with HCC beyond the Milan criteria exhibited a high rate of HCC recurrence after hepatectomy.Compared to the ETV therapy,TDF administration significantly lowered the risk of HCC recurrence.

Safety of elective hepatectomy performed on weekend for patients with hepatocellular carcinoma

The incidence of hepatocellular carcinoma(HCC)is increasing year by year.The International Agency for Research on Cancer(IARC)reported that more than 630,000 people were diagnosed with HCC in 2020 and more than half of new cases of HCC were in China.The increasing number of patients overburdens the medical system and the waiting time before the surgery becomes longer and longer.This poses a new challenge to the applicability of traditional surgical management model.The elective surgery performed on weekend could provide an efficient solution to the problem.However,the safety of weekend surgery is controversial because surgical patients have a higher severe complication incidence and postoperative mortality on the weekend compared to the weekday(ie,weekend effect).

Examining the Factors Affecting Personal Income： An Empirical Study Based on Survey Data in Chinese Cities

This paper empirically analyzes the factors affecting personal income in urban China using survey data of the ＂Preference and Life Satisfaction Survey＂ conducted by the Global COE project of Osaka University from 2009 to 2013. We consider education level as an endogenous variable, and both ordinary least squares （OLS） regression and instrumental variable （IV） regression are performed. We find a number of factors, such as sex, age, education, and marriage that significantly affect personal income. In addition, differences between different occupations are also investigated.