Liver cancer is the sixth most common type of cancer worldwide and the third leading cause of cancer-related deaths.Approximately 30％of patients are diagnosed at an early stage of liver cancer,at which point cura-tive treatments such as partial liver resection and liver transplantation can be performed,leading to a median overall survival exceeding 60 months.However,most patients are diagnosed at an advanced stage of liver cancer,where treatment options often include targeted therapies and immune checkpoint inhibitors.Systemic treatments including immune checkpoint inhibitors play a crucial role in significantly improving the prognosis of patients with advanced liver cancer.Therefore,in-depth exploration of the immune microenvironment characteristics of liver cancer and ac-tive identification of biomarkers for immunotherapy are crucial to advancing and improving immunotherapy for liver cancer.

Objective To investigate the characteristic genes of Programmed cell death protein 1(PD-1)immuno-therapy sensitivity in Hepatocellular carcinoma(HCC).Methods The common differential genes in GSE202069 and ERP117672 data sets were investigated by Weighted Gene Co-expression Network Analysis(WGCNA)and Difference analysis,and the characteristic genes of PD-1 immunotherapy sensitivity were screened through Lasso regression.The expression levels of characteristic genes in HCC were predicted by GEPIA and Ualcan databases,and their expression was verified by real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR),Western blot(WB)and Immunohistochemistry(IHC).3-hydroxybutyrate dehydrogenase 1(BDH1)over-expressed cell line was constructed,followed by cell counting kit-8(CCK-8),EdU,cell scratches and Transwell experiment to investigate the effects of BDH1 on the proliferation,migration and invasion of HCC cells.Results Total of 118 common differentially expressed genes were identified in two datasets by WGCNA and differential anal-ysis.The characteristic genes associated with PD-1 immunotherapy sensitivity screened through Lasso regression in-cluding Flavin containing dimethylaniline monoxygenase 3(FMO3),Peroxisomal trans-2-enoyl-CoA reductase(PECR),BDH1,Solute carrier family 7 member 1(SLC7A1),Cytochrome b5 type A(CYB5A)and Phos-phoenolpyruvate carboxykinase 1(PCK1).Survival analysis showed that BDH1 was most associated with HCC(Overall survival:P＜0.001,Recurrence:P=0.007).GEPIA and Ualcan databases showed low expression of BDH1 in HCC tissues,while RT-qPCR,WB,and IHC further confirmed this.CCK-8,plate cloning assay,EdU staining,cell scratch,and Transwell experiments showed that compared with the Hep3 B pCDH group,overexpres-sion of BDH1 resulted in a decrease in the absorbance of HCC cells(t=4.766,P＜0.01),a decrease in the num-ber of clone formation(t=16.02,P＜0.000 1),a decrease in the proportion of proliferating cells(t=23.13,P＜0.000 1),a decrease in cell migration rate(t=25.28,P＜0.000 1),and a decrease in the number of small compartments(t=10.78,P=0.004).Conclusion BDH1 is a characteristic gene for observing the sensitivity of PD-1 immunotherapy in HCC patients.BDH1 could inhibit the proliferation,migration,and invasion ability of HCC cells in vitro.

Objective To investigate the role of Glutathione peroxidase 2(GPX2)in the occurrence and progres-sion of intrahepatic cholangiocarcinoma(ICC).Methods The Omicshare website was used to analyze GPX2 ex-pression levels in ICC.The expression levels in ICC were validated using quantitative real-time reverse transcription PCR(RT-qPCR),Western blot,and immunohistochemistry.Stable GPX2 knockdown and overexpression HuC-CT1 cell lines were constructed.The effects of GPX2 on ICC cell proliferation,migration,apoptosis,and epithelial-mesenchymal transition(EMT)were investigated using colony formation assays,cell counting kit-8(CCK-8)as-says,wound healing assays,Transwell assays,and flow cytometry.A mouse model of cholangiocarcinoma was con-structed to assess GPX2 expression in mouse cholangiocarcinoma tissues.Results Based on the analysis results from the Omicshare website,GPX2 was generally upregulated in intrahepatic cholangiocarcinoma(ICC)(P＜0.001).Western blot(P＜0.000 1),RT-qPCR(P＜0.001),and immunohistochemistry experiments showed that,compared to adjacent non-cancerous tissues,the expression of GPX2 was significantly elevated in ICC.When GPX2 was knocked down,the colony formation rate of cells decreased significantly(P＜0.01),and the prolifera-tion capacity was reduced(P＜0.001).Conversely,overexpression of GPX2 led to a significant increase in colony formation rate(P＜0.01)and enhanced proliferation capacity(P＜0.01).Results from wound healing and Tran-swell assays demonstrated that GPX2 knockdown slowed down cell wound healing(P＜0.01)and reduced migra-tion ability(P＜0.01).Additionally,GPX2 knockdown resulted in an increase in E-cadherin(P＜0.01)and a decrease in N-cadherin(P＜0.01)and Vimentin(P＜0.05).On the other hand,overexpression of GPX2 accel-erated wound healing(P＜0.05)and enhanced migration ability(P＜0.05),while E-cadherin expression de-creased(P＜0.05)and N-cadherin(P＜0.01)and Vimentin(P＜0.001)expression increased.Flow cytometry for apoptosis and Western blot experiments indicated that GPX2 knockdown increased the apoptosis rate(P＜0.001),decreased the expression of Bcl-2(P＜0.001),and increased the expression of BAX(P＜0.01).But overexpression of GPX2 reduced the apoptosis rate(P＜0.01),increased Bcl-2 expression(P＜0.000 1),and decreased BAX expression(P＜0.001).Finally,elevated levels of GPX2 were observed in a mouse model of cholangiocarcinoma.Conclusion GPX2 is highly expressed in human and mouse cholangiocarcinoma tissues,and it may enhance cholangiocarcinoma cell proliferation and migration,promote tumor cell EMT,and inhibit tumor cell apoptosis.

Objective To investigate the expression of urocanase domain containing 1(UROC1)in Hepatocellular carcinoma(HCC)and its effect on the development of HCC.Methods UROC1 expression and prognostic data in tumor and para-tumor tissues from protein mass spectrometry and TCGA database were analyzed.Immunohistochem-ical staining,real time quantitative polymerase chain reaction(qPCR)and Western blot experiments were utilized to verify the expression of UROC1 in HCC.The effect of UROC1 expression level on tumor differentiation was ana-lyzed by HE staining to determine the tumor differentiation level.Hep3B and LM3 cell lines stably overexpressing UROC1 and its mutants were constructed for in vitro phenotyping experiments.Effects of UROC1 on HCC cell pro-liferation and migration were explored by cell counting kit-8(CCK-8)assay,colony formation,EdU assay,scratch assay and Transwell assay.Results The results of database analysis showed that UROC1 was generally downregu-lated in HCC tissues,and patients in the UROC1 low-expression group had a worse prognosis.Immunohistochemi-cal staining and scoring,qPCR and Western blot experiments verified the low expression of UROC1 in HCC.Im-munohistochemical staining of tumor tissues with different differentiation levels demonstrated that the poorer the dif-ferentiation of HCC tissues,the lower the expression level of UROC1.CCK-8,colony formation and EdU assays suggested that overexpression of UROC1 inhibited the proliferation of HCC cells.The results of scratch and Tran-swell assays showed that overexpression of UROC1 inhibited the migration of HCC cells.However,the results of the above experimental phenotypes after active site mutation converged with those of the control group.Conclusion UROC1 is lowly expressed in HCC tissues,and overexpression of UROC1 in HCC cells may inhibit the ability of cell proliferation and migration.

Objective To investigate the effect of N-myc downstream regulatory gene 1(NDRG1)on Hepatocellu-lar carcinoma(HCC)and whether NDRG1 affects the sensitivity of HCC to Sorafenib.Methods The expression level of NDRG1 in HCC was predicted by TCGA database,and verified by Western blot(WB)and Immunohisto-chemistry(IHC).NDRG1 knockout cell lines were constructed,followed by cell counting kit-8(CCK-8),EdU,cell scratches and Transwell experiment to investigate the effects of NDRG1 and its combination with Sorafenib on the proliferation,migration and invasion of HCC cells.In addition,flow cytometry was used to detect the apoptosis of HCC cells.The effect of NDRG1 and Sorafenib on HCC tumor formation was studied in vivo by subcutaneous tumor bearing in nude mice.WB and IHC were used to determine the pathway regulating the sensitivity of HCC to Sorafenib.Results WB and IHC confirmed that NDRG1 is highly expressed in HCC,consistent with the results of TCGA data.Tumor functional experiments showed that NDRG1 knockout or Sorafenib stimulation weakened the proliferation,migration,and invasion ability of HCC cells,and increased tumor cell apoptosis.However,NDRG1 knockout combined with Sorafenib further weakened the proliferation,migration and invasion ability of HCC cells,and further increased tumor cell apoptosis(P＜0.000 1).Mouse subcutaneous tumor model showed that NDRG1 knockout or Sorafenib stimulation reduced the tumor volume and quality,while NDRG1 knockout combined with Sorafenib further reduced the tumor volume and quality(P＜0.000 1).The WB and IHC results indicated that NDRG1 knockout combined with Sorafenib could reduce the phosphorylation level of Erk1/2.Conclusion NDRG1 is highly expressed in HCC,which promotes the proliferation,migration and invasion of HCC cells,and restricts apoptosis.NDRG1 knockout enhances HCC sensitivity to Sorafenib by reducing ERK signaling pathway phosphorylation.

Objective To investigate the mechanism by which human hypoxia up-regulation 1(HYOU1)regulates pancreatic cancer development.Methods Bioinformatic and immunohistochemical staining analyses of HYOU1 ex-pression level in pancreatic tumor tissues,normal and paracancerous tissues and its correlation with patients'surviv-al.Quantitative real-time PCR(qPCR)and Western blot were used to clarify the expression level of HYOU1 in a number of human pancreatic ductal adenocarcinoma cell lines and a normal human pancreatic ductal cell line.Two cell lines with the highest expression levels of HYOU1,BXPC-3 and Panc-1,were selected to knock out HYOU1 by CRISPR-Cas9,and then cell survival,proliferation and migration of these cells were examined by cell counting kit-8(CCK-8),colony formation assay and wound healing experiment separately,as well as apoptosis was detected by flow cytometry.Subsequently,the protein levels of the PI3K/Akt signaling including PI3K,p-PI3K,Akt,and p-Akt were detected by Western blot in parental and HYOU1-ablated BXPC-3 and Panc-1 cells.Cell proliferation was also examined in HYOU1-ablated cells after treatment of recilisib,an activator of the PI3K/Akt pathway.Results The expression of HYOU1 in pancreatic tumor tissues was significantly higher than that in normal tissues,and the patients with high expression of HYOU1 had a much shorter survival compared to the patients with low HYOU1(P＜0.01).Immunohistochemical staining of pancreatic cancer specimens showed that the expression of HYOU1 was higher in tumor tissues than in paracancerous tissues(P＜0.01).The mRNA and protein levels of HYOU1 were higher in all pancreatic cancer cell lines compared to the human normal pancreatic ductal cell(P＜0.001,P＜0.01).HYOU1 ablation inhibited BXPC-3 and Panc-1 cells survival,proliferation and migration,and promoted early cell apoptosis.In addition,loss of HYOU1 decreased PI3K/Akt signaling activity,whereas the PI3K/Akt ac-tivator Recilisib reversed the effects of HYOU1 ablation on cell survival and proliferation.Conclusion HYOU1 promotes pancreatic cancer progression by activating the PI3K/Akt signaling pathway.

Primary liver cancer, especially hepatocellular carcinoma, poses a serious threat to the life and health of the Chinese people. Given the insidious onset of liver cancer, less than 30% of hepatocellular carcinoma patients are considered for radical treatment at the initial diagnosis. Systemic anti-tumor therapy plays an important role in the treatment of advanced hepatocellular carcinoma. Immunotherapy of hepatocellular carcinoma has developed rapidly, and an increasing number of immunotherapy drugs, which can better control the progress of hepatocellular carcinoma and prolong the survival of patients, have become first- and second-line treatment options. This article reviews briefly the progress of immunotherapy for hepatocellular carcinoma in recent years.

Objective:To explore the risk factors of acute kidney injury(AKI)after liver transplantation(LT), examine its prognostic impact and construct a clinical prediction model.Methods:Clinical data are retrospectively reviewed for 220 LT recipients.They are divided into two groups of AKI(93 cases)and non-AKI(127 cases)according to the occurrence of AKI post-LT.Clinical data of two groups are compared.The variables with statistically significant inter-group differences in univariate analysis are included for multivariate analysis for obtaining the independent risk factors for AKI post-LT.Then the independent risk factors are employed for fitting a prediction model and a visual nomogram is constructed.At the same time, discrimination and calibration of the prediction model are evaluated.Extubation time, length of intensive care unit(ICU)stay, continuous renal replacement therapy(CRRT)rate, length of hospital stay, in-hospital mortality, estimated glomerular filtration rate(eGFR)at discharge, incidence of chronic renal failure(CRF)and readmission times are compared between two groups.Survival analysis is also performed between AKI and non-AKI groups and AKI 0/1 and AKI 2/3 stages.Results:The incidence of AKI post-LT is 42.3%.Age( OR=1.036, 95% CI: 1.001~1.073), preoperative serum creatinine level( OR=1.030, 95% CI: 1.011~1.049), platelet count( OR=0.992, 95% CI: 0.985~0.999), Child-Pugh class C( OR=2.678, 95% CI: 1.031~6.952), postoperative abdominal infection( OR=2.271, 95% CI: 1.120~4.603)and abdominal hemorrhage( OR=3.869, 95% CI: 1.016~14.72)are independent risk factors for AKI post-LT.The AUC/C-index of nomogram prediction model is 0.789 with a Brier score of 0.183, showing decent discrimination and calibration.According to the nomogram score, the recipients with a risk of AKI>50% are included into high-risk group while those with a risk of AKI<50% into low-risk group.Postoperative survival of low-risk group is better than that of high-risk group( P<0.001).Compared with non-AKI group, AKI group had a later extubation time( P=0.003), a longer length of ICU stay( P<0.001)and hospital stay( P=0.001), a higher rate of CRRT usage( P<0.001)and in-hospital mortality( P<0.001), a lower eGFR at discharge( P<0.001)and a higher incidence of CRF( P<0.001).Postoperative survival of non-AKI group was better than that of AKI group( P=0.048).Postoperative survival of patients with AKI 0/1 is better than that of those with AKI 2/3( P=0.002). Conclusions:Advanced age, high preoperative serum creatinine, low preoperative platelet, poor preoperative liver function, postoperative abdominal infection and abdominal hemorrhage may elevate the risks of AKI post-LT.And the nomogram prediction model based upon the above risk factors has a high value of clinical application.

The caudate lobe of liver is anatomically divided into three parts: Spiegel portion, inferior vena cava portion and caudate process. The caudate lobe of the liver is located in the dorsal side of the liver, adjacent to the inferior vena cava, the three hepatic veins, and the left and right portal veins. The location of the caudate lobe depends on the location of anatomical landmarks and the location of staining, especially negative staining techniques. The left approach is suitable for Spiegel resection, and the right approach is suitable for paracentral resection of the inferior vena cava and caudate process. The dorsal approach and anterior approach combined with other approaches can achieve complete caudate resection. This article showed the combination of multimodal approach with total caudate lobectomy, partial caudate lobectomy and laparoscopic caudate lobectomy.