Objective To predict the mechanism of herbal couple Curcumae Rhizoma-Sparganii Rhizoma(CR-SR)in modulation of angiogenesis against lung cancer based on network pharmacology and molecular docking technology,and validate by zebrafish model.Methods The active ingredients and potential targets for anti-lung cancer and antiangiogenesis of CR and SR were screened by network pharmacology.The targets were intersected with those screened from the OMIM database and GeneCards database for lung cancer and antiangiogenesis.Herbal couple-lung cancer and herbal couple-antiangiogenesis of protein-protein interaction(PPI)network was constructed by taking intersecting targets to screen the common and core targets of the herbal couple in lung cancer and anti-angiogenesis.Herbal couple-lung cancer and herbal couple-antiangiogenesis of Gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analyses were performed by Metascape database.The binding ability and the amino acid residues involved of core targets to major components were evaluated by molecular docking technique.In vitro,CCK-8 method was applied to investigate the effects of herbal couple and single drugs on the cell viability of human umbilical vein endothelial cells(HUVECs).Zebrafish embryos were randomly divided into blank control group,different concentration of drug pairs and single drug group,and positive drug control group,and the number of intersegmental vessels of zebrafish in each group was counted after 72 hour.The mRNA expression levels of angiogenesis-related genes,VEGFA,VEGFR2,VEGFR3,EGFR,etc.,were detected by qRT-PCR.Results 106 herbal couple-lung cancer common targets and 130 herbal couple-antiangiogenesis common targets were screened by network pharmacology.Meanwhile,85 of targets were identical.GO function enrichment analyses of herbal couple-lung cancer resulted in 1648 GO analysis entries,KEGG pathway enrichment analyses resulted in 186 signaling pathways.GO function enrichment analyses herbal couple-antiangiogenesis resulted in 1844 GO analysis entries,KEGG pathway enrichment analyses resulted in 188 signaling pathways.The molecular docking results showed a better affinity between the target and the components,and the forces between them mainly included hydrogen bonding and hydrophobic interactions.In vitro cellular experiments demonstrated that the two drugs were used as a drug pair to enhance the inhibitory effect on the cell viability of HUVECs.The zebrafish experiments indicated that the toxicity order of herbal couple and single drugs was CR>CR-SR>SR.The results of transgenic zebrafish vascular fluorescence model confirmed that CR-SR and single drugs had anti-angiogenic activity,with the anti-angiogenic activity order of herbal couple and single drugs was CR-SR>SR>CR.The results of qRT-PCR showed that CR-SR drug pairs and single drugs significantly reduced the expression levels of angiogenesis-related genes VEGFA,VEGFR2,EGFR,MMP9,etc.,and had anti-angiogenic effects.Conclusion CR-SR and single drugs had anti-lung cancer effects on multiple identical targets and regulated multiple identical signaling pathways,and their combination had a synergistic effect.The treatment of lung cancer may be through the regulation of angiogenesis-related target VEGFA,VEGFR2,EGFR,etc.,in order to play an anti-angiogenic effect.

Objective To predict the mechanism of herbal couple Curcumae Rhizoma-Sparganii Rhizoma(CR-SR)in modulation of angiogenesis against lung cancer based on network pharmacology and molecular docking technology,and validate by zebrafish model.Methods The active ingredients and potential targets for anti-lung cancer and antiangiogenesis of CR and SR were screened by network pharmacology.The targets were intersected with those screened from the OMIM database and GeneCards database for lung cancer and antiangiogenesis.Herbal couple-lung cancer and herbal couple-antiangiogenesis of protein-protein interaction(PPI)network was constructed by taking intersecting targets to screen the common and core targets of the herbal couple in lung cancer and anti-angiogenesis.Herbal couple-lung cancer and herbal couple-antiangiogenesis of Gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analyses were performed by Metascape database.The binding ability and the amino acid residues involved of core targets to major components were evaluated by molecular docking technique.In vitro,CCK-8 method was applied to investigate the effects of herbal couple and single drugs on the cell viability of human umbilical vein endothelial cells(HUVECs).Zebrafish embryos were randomly divided into blank control group,different concentration of drug pairs and single drug group,and positive drug control group,and the number of intersegmental vessels of zebrafish in each group was counted after 72 hour.The mRNA expression levels of angiogenesis-related genes,VEGFA,VEGFR2,VEGFR3,EGFR,etc.,were detected by qRT-PCR.Results 106 herbal couple-lung cancer common targets and 130 herbal couple-antiangiogenesis common targets were screened by network pharmacology.Meanwhile,85 of targets were identical.GO function enrichment analyses of herbal couple-lung cancer resulted in 1648 GO analysis entries,KEGG pathway enrichment analyses resulted in 186 signaling pathways.GO function enrichment analyses herbal couple-antiangiogenesis resulted in 1844 GO analysis entries,KEGG pathway enrichment analyses resulted in 188 signaling pathways.The molecular docking results showed a better affinity between the target and the components,and the forces between them mainly included hydrogen bonding and hydrophobic interactions.In vitro cellular experiments demonstrated that the two drugs were used as a drug pair to enhance the inhibitory effect on the cell viability of HUVECs.The zebrafish experiments indicated that the toxicity order of herbal couple and single drugs was CR>CR-SR>SR.The results of transgenic zebrafish vascular fluorescence model confirmed that CR-SR and single drugs had anti-angiogenic activity,with the anti-angiogenic activity order of herbal couple and single drugs was CR-SR>SR>CR.The results of qRT-PCR showed that CR-SR drug pairs and single drugs significantly reduced the expression levels of angiogenesis-related genes VEGFA,VEGFR2,EGFR,MMP9,etc.,and had anti-angiogenic effects.Conclusion CR-SR and single drugs had anti-lung cancer effects on multiple identical targets and regulated multiple identical signaling pathways,and their combination had a synergistic effect.The treatment of lung cancer may be through the regulation of angiogenesis-related target VEGFA,VEGFR2,EGFR,etc.,in order to play an anti-angiogenic effect.

Objective:To investigate the status of hepatitis E virus infection in unpaid blood donors in Wenzhou.Methods:A total of 5 241 blood samples were selected from donors who successfully donated blood between October 2022 and March 2023. These samples were tested for hepatitis E virus (HEV) through HEV-IgG antibodies, HEV-IgM antibodies, and nucleic acid tests (HEV RNA). Samples that tested positive for HEV RNA underwent quantitative detection and sequencing. Data regarding the blood donors were collected, the positive rates of each test result were recorded, and the HEV infection status among different populations was analyzed.Results:The positive rate of the HEV-IgG antibody test was 23.60% (1 237/5 241), while the positive rate of the HEV-IgM antibody test was 0.29% (15/5 241). The positive rate of the HEV RNA test was 0.08% (4/5 241). The positive rate of the HEV-IgG test increased with age ( χ2 = 379.493, P < 0.05), and males had a higher positive rate than females ( χ2 = 5.12, P < 0.05). Both the positive rates of HEV-IgG and HEV-IgM tests increased as the number of blood donations increased ( χ2 = 40.87, 8.30, both P < 0.05). Furthermore, the positive rates were significantly higher in 2023 than in 2022 ( χ2 = 51.03, 13.35, both P < 0.05), and they were also significantly higher in spring than in winter ( χ2 = 51.03, 13.35, both P < 0.05). Only one HEV RNA-positive blood donor tested positive for both HEV-IgG and HEV-IgM antibodies, while the remaining three donors tested negative. Only one case was genotyped, and it belonged to genotype Ⅳ. Conclusion:The incidence of HEV infection among unpaid blood donors in Wenzhou of Zhejiang Province is relatively high, with the prevalent HEV genotype being type Ⅳ. Characteristics such as sex, age, and the number of blood donations can be utilized as auxiliary screening reference indicators.

Ferroptosis is a non-apoptotic regulated cell death caused by iron accumulation and subsequent lipid peroxidation. Currently, the therapeutic role of ferroptosis on cancer is gaining increasing interest. Baicalin an active component in

DNA methylation is a prevalent epigenetic modification in vertebrates, and it has been shown to be involved the regulation of gene expression and embryo development. However, it remains unclear how DNA methylation regulates sexual development, especially in species without sex chromosomes. To determine this, we utilized zebrafish to investigate DNA methylation reprogramming during juvenile germ cell development and adult female-to-male sex transition. We reveal that primordial germ cells (PGCs) undergo significant DNA methylation reprogramming during germ cell development, and the methylome of PGCs is reset to an oocyte/ovary-like pattern at 9 days post fertilization (9 dpf). When DNA methyltransferase (DNMT) activity in juveniles was blocked after 9 dpf, the zebrafish developed into females. We also show that Tet3 is involved in PGC development. Notably, we find that DNA methylome reprogramming during adult zebrafish sex transition is similar to the reprogramming during the sex differentiation from 9 dpf PGCs to sperm. Furthermore, inhibiting DNMT activity can prevent the female-to-male sex transition, sug-gesting that methylation reprogramming is required for zebrafish sex transition. In summary, DNA methylation plays important roles in zebrafish germ cell development and sexual plasticity.

Objective To study the effects of eliminating free radical and increasing antioxidative capacity of glutathione peroxidase 1(GPX1) on PC12 cells. MethodsGPX1 recombinant plasmid and Plncxplasmid were transfected into PC12 cells and PC12 cells highly-expressing GPX1 stably were sieved by G418 solution. PC12 ceils were treated with different concentrations of amyloid β-protein (Aβ25-35) for 48 h, to decide the optimal concentration of Aβ25-35 and construct ideal cell model. GPX1/pLNCX/PC12 group, pLNCX/ PC12 group and PC12 group were treated with optimal concentration of Aβ25-35 ,respectively for 48 h, and their absorbance (A) value by MTT conversion was compared among three groups.ResultsCell clone highly expressing GPX1 stably were obtained by G418 selection. The increment of cell inhibition ratio was 24.7 ％ after 20 μmol/L Aβ25-35 treatment for 48 h, compared with control group (P＜0.01). Thus the optimization concentration of Aβ25-35 was 20 μmol/L. After treatment with 20 μmol/L for 48 h, the A value was significantly higher in GPX1/pLNCX/PC12 group than in pLNCX/ PC12 group and in PC12 cells group(0.53±0. 02 vs. 0.44±0.02;0.53±0.02 vs. 0.39±0.07, P＜0.01). Conclusions Transfection of GPX1 recombinant plasmid may protect cell against injury from free radical and reverse the decrease of PC12 cell survival rate impaired by Aβ25-35.