Objective This study aimed to investigate the expression and prognostic significance of Tubulin Beta 2A Class IIa(TUBB2A)in hepatocellular carcinoma(HCC)and to explore its roles in modulating malignant behavior of HCC cells under hypoxia.Methods The expression of TUBB2A in HCC tissues and its correlation with patient prognosis were analyzed using data from The Cancer Genome Atlas Program(TCGA)and validated by Real-time quantitative PCR(qPCR).HepG2 and Hep3B cells were cultured under both normoxia and hypoxia,and followed by assessment of mRNA and protein levels of TUBB2A by qPCR and Western blot.Stable TUBB2A overexpressing and knockout cell models(HepG2 and Hep3B)were established using lentiviral infection technology.The biological function of TUBB2A in HCC cell proliferation,mobility,and invasion of HCC cells under hypoxia were assessed by CCK-8 assay,colony formation assay,and Transwell migration and invasion assays.Western blot was performed to examine the impact of TUBB2A on the expression of key glycolytic proteins,including glucose transporter 1(GLUT1),pyruvate kinase M2(PKM2),and lactate dehydrogenase A(LDHA)in HCC cells under hypoxia.Results TCGA analysis revealed that TUBB2A expression is highly upregulated in HCC tissues compared to that in adjacent tissues and TUBB2A expression is associated with poorer overall survival(P<0.01).The mRNA expression of TUBB2A in HCC tissues from 30 HCC patients was higher than that in corresponding adjacent tissues,and TUBB2A expression is positively associated with the Ⅰ and Ⅳ clinic stage and AFP level of HCC patients(P<0.05).Hypoxia significantly increased TUBB2A mRNA and protein expression in HepG2 and Hep3B cells.Functional studies demonstrated that TUBB2A overexpression under hypoxia enhanced HCC cells proliferation,mobility and invasion,as well as increased the expression of GLUT1、PKM2 and LDHA(P<0.05 or P<0.01).On the contrary,TUBB2A knockout under hypoxia showed opposite effects,suppressed these malignant phenotypes and downregulated glycolytic markers(P<0.05 or P<0.01).Conclusions TUBB2A is significantly overexpressed in HCC and it is closely associated with poor prognosis.Our findings demonstrated that TUBB2A promotes proliferation,migration,invasion and glycolysis in HCC cells under hypoxia,suggesting its potential as a prognostic biomarker and therapeutic target of HCC.

Objective This study aimed to investigate the expression and prognostic significance of Tubulin Beta 2A Class IIa(TUBB2A)in hepatocellular carcinoma(HCC)and to explore its roles in modulating malignant behavior of HCC cells under hypoxia.Methods The expression of TUBB2A in HCC tissues and its correlation with patient prognosis were analyzed using data from The Cancer Genome Atlas Program(TCGA)and validated by Real-time quantitative PCR(qPCR).HepG2 and Hep3B cells were cultured under both normoxia and hypoxia,and followed by assessment of mRNA and protein levels of TUBB2A by qPCR and Western blot.Stable TUBB2A overexpressing and knockout cell models(HepG2 and Hep3B)were established using lentiviral infection technology.The biological function of TUBB2A in HCC cell proliferation,mobility,and invasion of HCC cells under hypoxia were assessed by CCK-8 assay,colony formation assay,and Transwell migration and invasion assays.Western blot was performed to examine the impact of TUBB2A on the expression of key glycolytic proteins,including glucose transporter 1(GLUT1),pyruvate kinase M2(PKM2),and lactate dehydrogenase A(LDHA)in HCC cells under hypoxia.Results TCGA analysis revealed that TUBB2A expression is highly upregulated in HCC tissues compared to that in adjacent tissues and TUBB2A expression is associated with poorer overall survival(P<0.01).The mRNA expression of TUBB2A in HCC tissues from 30 HCC patients was higher than that in corresponding adjacent tissues,and TUBB2A expression is positively associated with the Ⅰ and Ⅳ clinic stage and AFP level of HCC patients(P<0.05).Hypoxia significantly increased TUBB2A mRNA and protein expression in HepG2 and Hep3B cells.Functional studies demonstrated that TUBB2A overexpression under hypoxia enhanced HCC cells proliferation,mobility and invasion,as well as increased the expression of GLUT1、PKM2 and LDHA(P<0.05 or P<0.01).On the contrary,TUBB2A knockout under hypoxia showed opposite effects,suppressed these malignant phenotypes and downregulated glycolytic markers(P<0.05 or P<0.01).Conclusions TUBB2A is significantly overexpressed in HCC and it is closely associated with poor prognosis.Our findings demonstrated that TUBB2A promotes proliferation,migration,invasion and glycolysis in HCC cells under hypoxia,suggesting its potential as a prognostic biomarker and therapeutic target of HCC.

Objective:To screen T-cell exhaustion-related signature genes as the prognostic marker for osteosarcoma and establish a prognostic model for osteosarcoma patients based on weighted gene co-expression network analysis(WGCNA)and Least absolute shrinkage and selection operator(LASSO)-COX regression analysis. Methods:GSE21257 dataset was downloaded from Gene Expression Omnibus(GEO)database for the establishment of the prognostic model for osteosarcoma.4 T-cell exhaustion-related gene sets were downloaded from The Molecular Signatures Database(MisgDB)and their enrichment scores in GSE21257 samples were calculated by single sample gene set enrichment analysis(ssGSEA).WGCNA was carried out to screen the gene module that is highly associated with T-cell exhaustion based on ssGSEA results followed by GO(Gene Ontology)and KEGG(Kyoto Encyclopedia of Genes and Genomes)analysis of the biological processes and signaling transduction pathways that those genes are involved in.The signature genes that are highly associated with the prognosis of osteosarcoma patients were obtained through LASSO-COX regression and a prognostic model was established based on these signature genes.Osteosarcoma-related expression profile data from the GSE21257 and TAEGET datasets on XENA were downloaded from the Gene Expression Omnibus.Clinical information for the training and validation sets was obtained.T-cell exhaustion-related genes were screened using a weighted correlation network analysis.Realtime fluorescence quantitative PCR,COX regression analysis,external dataset and nomogram were used to evaluate the reliability and accuracy of the prognostic model.A immunotherapy-related dataset was used to assess the efficacy of this prognostic model for the prediction of patients'responses to immunotherapy. Results:Analysis results based on the ssGSEA scores showed that T-cell exhaustion-related genes were related to the metastasis and age of osteosarcoma patients.Many T-cell exhaustion-related genes were found to be differentially expressed in metastatic and non-metastatic osteosarcoma patients.1 256 T-cell exhaustion-related genes were identified through WGCNA and these candidate markers were mainly distributed in structures like secretory granule membranes and endocytic vesicles and were involved in T-cell activation.COX regression analysis screened 68 significant prognostic markers out of the 1 256 genes,and 12 signature genes were further confirmed with LASSO-COX regression analysis.A prognostic model was established based on the 12 signature genes.Results of real-time fluorescence quantitative PCR showed a similar trend in the expression of most of the signature genes in different osteosarcoma cell lines.COX regression analysis of the internal and external datasets verified that the risk score calculated with the prognostic model was an independent prognostic factor for osteosarcoma patients,and high-risk score was associated with poor prognosis of the patients.Receiver operating characteristic(ROC)curves demonstrated excellent prognostic efficacy of the model.Nomogram analysis verified the prognostic model is highly accurate and reliable in predicting the prognosis of osteosarcoma patients.Analysis using the immunotherapy-related dataset indicated that this prognostic model could also be used to predict patients'responses to immunotherapy. Conclusion:The 12 signature gene(CD300LB,TRO,SNX3,VENTX,PPM1M,DOT1L,CDC37,NAT9,TRMT1,PPP1R3C,CHTF18 and NSUN5)-based prognostic model can effectively predict the prognosis and responses to immune check-point inhibitors for osteosarcoma patients,which may provide evidence for the prediction of prognosis as well as the selection of immunotherapy plans in clinical practice.

Objective To observe the clinical therapeutic effects and evaluate the security of Huganjiexian decoction combined with conventional therapy on hepatic cirrhosis.MethodsBy the randomized and prospective study method,34 patients with liver cirrhosis were divided into experimental group and control group.The experimental group was treated with Huganjiexian decoction combined with conventional therapy while the control group was treated with conventional therapy alone.Patients in both groups were treated six months.At the beginning and 6 months after treatment,levels of alanine transaminase (ALT),aspartate transaminase (AST),albumin (ALB),albumin/globulin (A/G),total bilirubin (TBiL),blood urea nitrogen (BUN),serum creatinine (Scr) were determined.Results Levels ofALT、AST、TBiL decreased in both groups after being treated for six months,and the differences of downward trend of the experimental group were more significant than control group (F=36.63,40.31,38.65,P＜0.05).Levels ofALT、AST、TBiL of the experimental group were lower than those of control group significantly (F=8.67,7.62,4.36,P＜0.05 ).The A/G raised in both groups after treatment,and the upward trend of the experimental group was greatly different from that of control group (F=24.10,P＜0.05),the value of A/G of the experimental group was higher than that of control group (F=4.78,P＜0.05).The ALB raised in both groups after treatment,while the upward trend of the experimental group was no different from that of control group (F=0.89,P＞ 0.05).Thevalue of ALB had no significant changes in both groups (F=3.15,P＞0.05).Conclusion Huganjiexian decoction possessed therapeutic effect on hepatic cirrhosis,it had no obvious toxicity and side