Objective This study aims to investigate the effects of transcatheter arterial chemoembolization(TACE)using EqualSpheres,CalliSpheres,and Lipiodol loaded with idarubicin on VX2 liver cancer in rabbits.Methods Twelve New Zealand white rabbits were randomly assigned to three groups:EqualSpheres group,CalliSpheres group,and Lipiodol group(n=4 per group).The VX2 liver cancer animal model was successfully established through ultrasound-guided percutaneous puncture.EqualSpheres,CalliSpheres,and Lipiodol were employed as embolization agents loaded with idarubicin for the embolization procedure.Peripheral blood samples were collected at intervals of 5 minutes,0.5,1,4,12 and 24 hours following embolization and were centrifuged to obtain serum.At 24 hours post-TACE treatment,the rabbits in both the experimental and control groups were euthanized,and both liver cancer tissues and normal liver tissues were collected.UPLC-MS/MS was used to measure the drug concentration of idarubicin in peripheral blood and tissue samples,and Graphpad software was used to construct drug concentration-time curves in peripheral blood.The pharmacokinetic curves were constructed to evaluate the dynamic in vivo distribution characteristics.Results The average drug concentration in the EqualSpheres group(920.06 ng/mL)was significantly higher than that in both the CalliSpheres group(79.47 ng/mL)and the Lipiodol group(118.71 ng/mL).However,the average drug concentration in normal liver tissue of all the three groups was lower,and the difference was not statistically significant.The peripheral blood drug concentration of the three groups decreased at 5 minutes post-TACE and increased over the next 24 hours.The average blood concentration curve of EqualSpheres group increased more steadily compared to the CalliSpheres group and the Lipiodol group.The Cmax of the Lipiodol group was reached at 0.5 hours,measuring 11.54 ng/mL.Both the CalliSpheres group and the EqualSpheres group achieved their Cmax at 5 minutes,with values of 7.82 and 8.36 ng/mL,respectively.Conclusion EqualSpheres loaded with idarubicin achieve a high drug concentration at the tumor site while maintaining a low concentration in peripheral blood over 24 hours.This study demonstrates the stable drug release capability of idarubicin-loaded EqualSpheres.

Breast cancer remains a leading cause of mortality in women worldwide.Triple-negative breast cancer(TNBC)is a particularly aggressive subtype characterized by rapid progression,poor prognosis,and lack of clear therapeutic targets.In the clinic,delineation of tumor heterogeneity and development of effective drugs continue to pose considerable challenges.Within the scope of our study,high hetero-geneity inherent to breast cancer was uncovered based on the landscape constructed from both tumor and healthy breast tissue samples.Notably,TNBC exhibited significant specificity regarding cell prolif-eration,differentiation,and disease progression.Significant associations between tumor grade,prog-nosis,and TNBC oncogenes were established via pseudotime trajectory analysis.Consequently,we further performed comprehensive characterization of the TNBC microenvironment.A crucial epithelial subcluster,E8,was identified as highly malignant and strongly associated with tumor cell proliferation in TNBC.Additionally,epithelial-mesenchymal transition(EMT)-associated fibroblast and M2 macrophage subclusters exerted an influence on E8 through cellular interactions,contributing to tumor growth.Characteristic genes in these three cluster cells could therefore serve as potential therapeutic targets for TNBC.The collective findings provided valuable insights that assisted in the screening of a series of therapeutic drugs,such as pelitinib.We further confirmed the anti-cancer effect of pelitinib in an orthotopic 4T1 tumor-bearing mouse model.Overall,our study sheds light on the unique characteristics of TNBC at single-cell resolution and the crucial cell types associated with tumor cell proliferation that may serve as potent tools in the development of effective anti-cancer drugs.

BACKGROUNDTo elucidate relationship between amino acid sequence of non-structural protein 5A (NS5A) and outcome of HCV (1 b) patients after interferon (IFNa) therapy.

METHODSSera of 24 patients were collected before, during and after IFNa therapy. Pretreatment RNA levels and the sequences of HCV NS5A interferon sensitivity determining region (ISDR) were determined. NS5A full-length sequences of 5 HCV isolates from 3 patients with different response types were also analyzed. Phylogenetic tree analysis and protein secondary structure prediction were undertaken.

RESULTSPretreatment RNA levels of sustained response group were significantly lower than that of non-response group and relapse group (4.50X104 copies/ml versus 1.82X107 copies/ml, P < 0.01).ISDR sequences of NS5A from pretreatment sera were compared with HCV-J strain (prototype). Thirteen of 24 isolates were wild type,11 of 24 were intermediate type and none of them was mutant type. 3 of 6 sustained responders were infected with wild-type isolates, the rest with intermediate type isolates. Phylogenetic tree based on NS5A full-length sequences classified 5 isolates with 3 different response types into 3 groups. Non-response isolates belonged to the same group as HCV-J. Secondary structure prediction of 5 isolates revealed significant differences existing in 2 255- 2 289. This region was partly overlapped with PKR-binding domain.

CONCLUSIONSLow HCV RNA levels in serum are associated with favorable outcome of IFNa therapy. ISDR sequence alone could not predict outcome of IFN treatment. Combination of determination of HCV RNA levels in serum with sequence analysis of PKR-binding domain may be helpful in predicting the efficacy of IFN therapy.

Amino Acid Sequence ; Antiviral Agents ; therapeutic use ; Hepacivirus ; drug effects ; genetics ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; RNA, Viral ; blood ; Viral Nonstructural Proteins ; genetics

Objective To study the correlation of replication efficiency and antigen expression of hepatitis B virus (HBV) strains in patient sera and transfected cells. Methods Quantification of five maternal serum HBV DNA was carried out using dot hybridization and polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA), hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels in the sera were determined by ELISA. Full-length genomes of HBV strains cloned from five pregnant women were separately used to transfect HepG2 cells. The levels of HBsAg and HBeAg in the supernatant of transfected cells were determined with Abbott EIA kits, the replication efficiency of intracelluar replicative intermediates and extracellular HBV DNA of viral particles was analyzed by Southern blot and hybridization. Results Intracellular and extracellular HBV replication and antigen expression of cloned HBV strains showed positive correlation tendency with the HBV DNA and antigen levels in serum samples. Conclusions Virus replication efficiency and expression of HBsAg and HBeAg by full-length HBV clones are in relative coincidence with the biological characteristics of HBV strains in patients. Cell transfection can be used to study the biological characteristics of individual isolates.