Objective:To investigate the effects of transhepatic arterial chemoembolization(TACE)combined with Endostar on CD4+/CD8+T cells,5-aminoketovalonate synthase 1(ALAS1)and HIPPO-YAP signaling pathway in patients with hepatocellular carcinoma(HCC).Methods:A total of 60 HCC patients admitted to Chengdu Third People's Hospital from January 2018 to December 2021 were enrolled and randomly divided into TACE treatment group(A)and TACE combined with Endostar treatment group(B),with 30 patients in each group.T lymphocyte,ALAS1 and indicators of HIPPO-YAP signaling pathway were observed in the two groups.Results:In group A,1 subject had complete remission,2 subjects had partial remission,2 subjects had stable remission,and 4 subjects had progress;in group B,2 subjects had complete remission,4 subjects had partial remission,4 subjects had stable remis-sion,and 1 subject had progress.The total effective rate in group B(33.33%)was significantly higher than that in group A(10.00%),with significant difference among groups(P=0.028).There were no significant differences in CD4+T cells,CD8+T cells and CD4+/CD8+T cells between the two groups before treatment(P=0.972,0.995,0.917).After treatment,level of CD4+T cells in the two groups was significantly increased(P<0.001),while level of CD8+T cells was significantly decreased(P<0.001).CD4+/CD8+T cells was significantly increased(P<0.001),and the changes in group B were significant compared with group A(P<0.001).There was no significant difference in content of ALAS1 between the two groups before treatment(P=0.975);after treatment,content of ALAS1 in liver cancer tissues of the two groups was significantly increased(P<0.001),which in group B was significantly higher than that in group A(P<0.05).Before treatment,there were no significant differences in mammalian STE20-like protein kinase 2(MST2),large tumor suppressor 1(LATS1)and Yes-associated protein(YAP)between the two groups(P=0.134,0.134,0.134).After treatment,mRNA relative expressions of MST2 and LATS1 were significantly increased(all P<0.001),while mRNA relative expression of YAP were significantly decreased(P<0.001),and changes of group B were significant compared with group A(all P<0.001).After treat-ment,there were 5 more cases of no adverse reactions in group B than in group A,and the total incidence of adverse reactions(16.67%)was significantly lower than that in group A(43.33%),with a significant difference between groups(P=0.024).Conclu-sion:TACE combined with Endostar has significant therapeutic effect on HCC patients,which can effectively regulate CD4+/CD8+T cells,promote ALAS1 secretion,and activate HIPPO-YAP signaling pathway.

Objective:To investigate the effects of transhepatic arterial chemoembolization(TACE)combined with Endostar on CD4+/CD8+T cells,5-aminoketovalonate synthase 1(ALAS1)and HIPPO-YAP signaling pathway in patients with hepatocellular carcinoma(HCC).Methods:A total of 60 HCC patients admitted to Chengdu Third People's Hospital from January 2018 to December 2021 were enrolled and randomly divided into TACE treatment group(A)and TACE combined with Endostar treatment group(B),with 30 patients in each group.T lymphocyte,ALAS1 and indicators of HIPPO-YAP signaling pathway were observed in the two groups.Results:In group A,1 subject had complete remission,2 subjects had partial remission,2 subjects had stable remission,and 4 subjects had progress;in group B,2 subjects had complete remission,4 subjects had partial remission,4 subjects had stable remis-sion,and 1 subject had progress.The total effective rate in group B(33.33%)was significantly higher than that in group A(10.00%),with significant difference among groups(P=0.028).There were no significant differences in CD4+T cells,CD8+T cells and CD4+/CD8+T cells between the two groups before treatment(P=0.972,0.995,0.917).After treatment,level of CD4+T cells in the two groups was significantly increased(P<0.001),while level of CD8+T cells was significantly decreased(P<0.001).CD4+/CD8+T cells was significantly increased(P<0.001),and the changes in group B were significant compared with group A(P<0.001).There was no significant difference in content of ALAS1 between the two groups before treatment(P=0.975);after treatment,content of ALAS1 in liver cancer tissues of the two groups was significantly increased(P<0.001),which in group B was significantly higher than that in group A(P<0.05).Before treatment,there were no significant differences in mammalian STE20-like protein kinase 2(MST2),large tumor suppressor 1(LATS1)and Yes-associated protein(YAP)between the two groups(P=0.134,0.134,0.134).After treatment,mRNA relative expressions of MST2 and LATS1 were significantly increased(all P<0.001),while mRNA relative expression of YAP were significantly decreased(P<0.001),and changes of group B were significant compared with group A(all P<0.001).After treat-ment,there were 5 more cases of no adverse reactions in group B than in group A,and the total incidence of adverse reactions(16.67%)was significantly lower than that in group A(43.33%),with a significant difference between groups(P=0.024).Conclu-sion:TACE combined with Endostar has significant therapeutic effect on HCC patients,which can effectively regulate CD4+/CD8+T cells,promote ALAS1 secretion,and activate HIPPO-YAP signaling pathway.

In recent years, the development of bispecific antibodies (bsAbs) has been rapid, with many new structures and target combinations being created. The boom in bsAbs has led to the successive issuance of industry guidance for their development in the US and China. However, there is a high degree of similarity in target selection, which could affect the development of diversity in bsAbs. This review presents a classification of various bsAbs for cancer therapy based on structure and target selection and examines the advantages of bsAbs over monoclonal antibodies (mAbs). Through database research, we have identified the preferences of available bsAbs combinations, suggesting rational target selection options and warning of potential wastage of medical resources. We have also compared the US and Chinese guidelines for bsAbs in order to provide a reference for their development.

Objective:To study the inhibitory effect of ezetimibe in an experimental model of human hepatoma cell line (HepaRG) infected with hepatitis B virus (HBV) positive human serum in vitro.Methods:Mature HepaRG cells were divided into a treatment group (received drugs) and a control group (did not receive drugs). In the ezetimibe prevention experiment, the cells in the treatment group was treated with drugs 2 h before infection and 24 h during infection. In the ezetimibe treatment experiment, the cells in the treatment group were treated with drugs for 6 ~ 10 days continuously after 24 hours of HBV infection. The expression of HBV DNA and intracellular cccDNA in the supernatant was detected by fluorescence quantitative PCR. Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) content in the cell supernatant were detected by chemiluminescence. Analysis of variance was used to compare the differences between multiple groups. Pairwise comparisons among groups were followed by t- test with normal distribution. P < 0.05 was considered as statistically significant. Results:Ezetimibe prevention experiment showed that compared with control group, the treatment group was added with 20, 60, and 100 μmol/L ezetimibe before and during infection, and the HBV DNA content in the supernatant 2 days before was significantly reduced ( P < 0.05) in the treatment group. Compared with the control group, the HBsAg expression level 2 days before was significantly reduced ( P < 0.05) with the addition of 60 μmol/L ezetimibe in the treatment group. Compared with the control group, the expression level of intracellular cccDNA was significantly reduced ( P < 0.05) after 10 days with the addition of 100μmol/L ezetimibe in the treatment group. Ezetimibe treatment experiment showed that cccDNA content in the cells were significantly lowered with the immediate addition of 60μmol/L ezetimibe 24 hours after infection for 10 days when compared to control group ( P < 0.05). Conclusion:Ezetimibe, as a cytosolic inhibitor, has a certain inhibitory effect on hepatitis B virus infection in both prevention and treatment experiment.

BACKGROUND:Liposomes as a new drug delivery system are characterized by few adverse reactions, no immunogenicity and biodegradation. Furthermore, methotrexate-loaded liposomes can significantly reduce drug toxicity and improve anti-tumor effect. OBJECTIVE:To prepare long-circulating methotrexate-loaded liposomes and to evaluate its antitumor activity in MG-63 in vitro. METHODS:The methotrexate-loaded liposomes were prepared using the film dispersion method, and the long-circulating ones were prepared using the post-insertion method. The initial concentrations of methotrexate were 9.1, 1.82, 0.364 g/L. The ultracentrifugation method and spin column method with Sephadex G-10 or G-50 as packing were employed to separate free drugs from the methotrexate-loaded liposomes. Their recovery, size, morphology, encapsulation efficiency and drug-to-lipid ratio were evaluated. The cytotoxity of the long-circulating methotrexate-loaded liposomes purified with ultracentrifugation method and spin column G-50 method under three dose levels (0, 1, 5, 25 mg/L) were determined by MTS method. RESULTS AND CONCLUSION:According to the recovery rates of three methods, the spin column G-50 method was considered as optimal for the long-circulating methotrexate-loaded liposomes. The long-circulating liposomes were spherical or el ipsoidal under transmission electron microscope, about 200 nm in size. At the certain initial concentration of methotrexate, the encapsulation efficiency and drug-to-lipid ratio of the liposomes purified using the spin column G-50 method were remarkably higher than those purified using the other two methods. At the same mass concentration, the cytotoxity of the liposomes purified with ultracentrifugation or spin column G-50 was significantly higher than that of free methotrexate, and furthermore, the cytotoxity of the liposomes purified with spin column G-50 was higher than that of the liposomes purified with ultracentrifugation method. To conclude, the long-circulating methotrexate-loaded liposomes show a higher antitumor activity than free methotrexate in MG-63 cel s in vitro, providing the basis for further investigation of its antitumor effect on human osteosarcoma in vivo.

Objective To construct a extracellular matrix-like collagen mimetic peptide-PEG hybrid hydrogel and to study the usage of this hydrogel in 3D culture of rabbit bone marrow mesenchymal stem cells (rBMSCs).Methods The hybrid hydrogel was synthesised by conjugating the cysteine at the end of the collagen mimetic peptide with the maleimine-modified multi-arm PEG.The circular dichroism spectra were used to characterize the triple helix structure and thermal stability of the collagen mimetic peptides.The rheology test and scanning electron microscopy were used to study the gelation process,mechanical strength and internal structure of the hydrogel.The rBMSCs were embedded in the hybrid hydrogel for 3D culture.The cell compatibility of the hydrogel and its effect on differentiation of the cells was studied.Results Collagen mimetic peptides could promote spontaneous formation of triple helix structure in the natural collagen,and the thermal transition temperature was 49.4 ℃.The formation process of the collagen mimetic peptides-PEG hybrid hydrogel was rapid,in which the porous network-like fibrous structure was formed.After the encapsulation of rBMSCs within the hydrogel for 24 h,most of the cells remained viable.Gene expression analysis showed that the hybrid hydrogel could affect the differentiation of rBMSCs.Conclusions The collagen mimetic peptide-PEG hybrid hydrogel possesses the characteristics of mild preparation condition,good mechanical strength and good cell compatibility,and is favorable to chondrocyte differentiation of rBMSCs.

Objective To improve the level of diagnosis and differentiation of renal benign mass with renal cell carcinoma(RCC),so as to lower the misdiagnosis rate.Methods This study included 9 cases of benign renal mass,whose age ranged from 30 to 76 years with a mean of 54 years and included 52 patients with RCC.Three subtypes of RCC were noted,including clear cell in 37 cases,papillary RCC in 10 cases and chromophobe RCC in 5 cases.Plain scan and three phase CT(corticomedullary,nephrographic and excretory phases)were done in all patients.The CT features of RCC and benign mass were compared.Results All the cases were underwent radical nephrectomy as RCC,while they were postoperatively diagnosed as benign renal mass.There were 4 cases of angiomyolipoma (AML)with minimal fat,two cases of oncocytoma,one case of leiomyoma,one case of inflammatory pseudotumor,and one case of cyst with hematoma and organization.Fifty-two cases of RCC showed homogenous or inhomogeous,equal,slightly lower,slightly higher or mixed density on unenhanced scan,inhomogenous obvious enhancement after administration of contrast media.And the most obviously enhanced portion of renal carcinomas were isodense or slightly hyperdense relative to adjacent renal cortex in corticomedullary phase.Conclusion CT is an important radiologic approach to diagnose and differentially diagnose malignant or benign kidney mass.For those patients with benign mass that is not a typical case on radiology,the preoperative needle biopsy or intraoperative frozen section pathological diagnosis is the key to avoid misdiagnose and mistake resection of the kidney,and choose the proper treatment approach to avoid unnecessary kidney radical resection.

Objective To investigate the influence factors on the graft hemodynamics after liver transplantation by CT perfusion(CTP).Methods Thirty three liver recipients received CT angiography (CTA)and CTP after liver transplantation.The cases would be excluded when their peak values of the aorta enhancement on time-density curves were out of 95%confidence level.The 95% confidence levels of the hepatic artery perfusion(HAP),portal vein perfusion(PVP),total liver perfusion(TLP)and hepatic perfusion index(HPI)were calculated based on the recipients without postoperative complications and named them as references to those with complication.Results Twenty nine recipients were enrolled in the study.15 of them had no postoperative complication while the other 14 had.The 95% confidence levels of HAP,PVP,TLP and HPI on the 15 recipients without complications were(0.1509-0.3183),the 14 cases with complications.HAP decreased in 7 cases,5 of them had hepatic artery stenosis and 3 of them had splenomegaly.HAP increased in 2 cases.both of them had portal vein stenosis.PVP decreased in 13 cases,8 of them had portal vein stenosis,portal vein thrombosis or occlusion,4 of them had splenorenal shunts and 2 of them had fatty liver.TLP decreased in 12 cases and coincident with PVP decreasing.Only 2 cases had HPI decreasing accompanied with HAP decreasing.Conclusion The hepatic blood perfusion through the hepatic artery and portal vein could be quantitatively measured non-invasively by CTP.The severity and the subtypes of the hepatic ischemia could be evaluated objectively,which is helpful for treatment guidance.