Objective:To investigate the related genes, signaling pathways and possible mechanisms of malignant transformation of human papillomavirus type 16 (HPV-16) immortalized cervical epithelial cell line H8.Methods:The malignant transformed H8 cell model was constructed, and the changes of cell invasion ability and cell migration ability of H8 cells after malignant transformation were detected by Transwell assay, and the changes of clone formation ability of H8 cells after malignant transformation were detected by plate clone formation assay. Total RNA was extracted from malignant transformed H8 cells and H8 cells, and the two groups of cells were sequenced by transcriptome using Illumina novaseq 6000 sequencing platform, differentially expressed genes (DEGs) were identified and analyzed, and Gene Ontology (GO) function enrichment analysis, Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis and protein-protein interaction were performed.Results:The invasion ability, migration ability and clone formation ability of malignant transformed H8 cells significantly increased as compared to H8 cells. A total of 203 differentially expressed genes were identified in H8 cells before and after malignant transformation, of which 98 were up-regulated and 105 down-regulated. GO enrichment analysis showed that DEGs were mainly involved in biological processes such as cellular processes, biological regulation, and metabolic processes. KEGG pathway enrichment analysis showed that DEGs were mainly enriched in alanine, aspartate and glutamate metabolic pathway, glycine, serine and threonine metabolism pathway, p53 signaling pathway and TGF-β signaling pathway, PI3K-Akt signaling pathway. PPI analysis screened 10 hub genes including DDIT3, TRIB3 and ASNS.Conclusions:Compared with H8 cells, malignant transformed H8 cells have a large number of differentially expressed genes and pathways at the transcriptional level, which could further provide new ideas for the mechanism of malignant transformation and carcinogenesis as well as finding new targets for the prevention of malignant transformation.

Objective To evaluate the protective effect of pterostilbene against ultraviolet B (UVB)-induced acute damage in HaCaT cells,and to explore related mechanisms.Methods The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazo1ium (MTS) assay and flow cytometry were performed to estimate the proliferative activity and the apoptosis and necrosis rate of HaCaT cells treated with different concentrations of pterostilbene respectively,so as to screen the non-toxic concentration of pterostilbene.HaCaT cells were randomly divided into several groups:normal control group receiving no treatment,UVB group irradiated with 57 mJ/cm2 UVB,3 pterostilbene groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours,3 pterostilbene + UVB groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours followed by UVB radiation.Western blot analysis was conducted to detect changes of the transcription factor NF-E2-related factor 2 (Nrf2) expression in cell nuclei and cytoplasm before and after the treatment with pterostilbene and UVB,quantitative PCR to determine the mRNA expression of catalase and superoxide dismutase in the HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to evaluate the activity of catalase and superoxide dismutase.Results MTS assay and flow cytometry showed that 2.44,4.88 and 9.75 μmol/L pterostilbene had non-toxic effect on HaCaT cells.The protein expression of Nrf2 in the nuclei and cytoplasm in the normal control group was 1.03 ± 0.08 and 1.04 ± 0.11 respectively.Compared with the normal control group,the protein expression of Nrf2 in the nuclei and cytoplasm experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups,and the UVB group showed similar protein expression of Nrf2 in the cytoplasm,but significantly increased protein expression of Nrf2 in the nuclei (1.77 ± 0.08,q =17.24,P ＜ 0.01).Compared with the normal control group and UVB group,the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups all showed significantly lower protein expression of Nrf2 in the cytoplasm (0.86 ± 0.10,0.87 ± 0.11 and 0.46 ± 0.11 respectively,all P ＜ 0.05),but significantly higher protein expression of Nrf2 in the nuclei (2.38 ± 0.11,2.57 ± 0.11 and 2.07 ± 0.13,all P ＜ 0.01).As qPCR showed,UVB radiation could significantly inhibit the mRNA expression of CAT (P ＜ 0.05),but had no obvious effect on the mRNA expression of SOD (P ＞ 0.05).The mRNA expression of CAT and SOD experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups compared with the normal control group (P ＞ 0.05).However,2.44,4.88 and 9.75 μmol/L pterostilbene could significantly reduce the inhibitory effect of UVB radiation on the mRNA expression of CAT (P ＜ 0.05) and up-regulate the mRNA expression of SOD in the pterostilbene + UVB groups (P ＜ 0.05).ELISA revealed that UVB radiation could inhibit the activity of CAT and SOD in the HaCaT cells (both P ＜ 0.001),while 2.44,4.88 and 9.75 μmol/L pterostilbene could reduce the inhibitory effect of UVB radiation on the activity of CAT and SOD (all P ＜ 0.05).However,the activity of CAT and SOD were still lower in the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups than in the normal control group (P ＜ 0.05).Conclusion Pterostilbene can prevent UVB-induced acute damage in HaCaT cells by activating the Nrf2 pathway and up-regulating the expression of the downstream antioxidant enzymes CAT and SOD.

Objective To explore the effect of tea polyphenols on the growth of human papillomavirus 16 (HPV16) subgenes-immortalized human cervical epithelial cells (H8 cells).Methods Cultured H8 cells were divided into 5 groups to be treated with 0 (control group),6.25,12.5,25 and 50 mg/L tea polyphenols respectively for 24,36,and 48 hours,and then cell counting kit-8 (CCK8)assay was performed to detect cell proliferation.After 24 hours of incubation,flow cytometry was conducted to detect cell apoptosis and cell cycle,and fluorescence microscopy to observe the morphology of apoptotic cells.Results After incubation with tea polyphenols at different concentrations for 24,36 and 48 hours,the proliferation of H8 cells was inhibited,and 12.5 mg/L tea polyphenols could inhibit the relative growth rate of H8 cells in a time-dependent manner.Flow cytometry showed that there was a significant difference in cell apoptosis rate among the 6.25-,12.5-,25-,50-mg/L tea polyphenols groups and the control group (52.62％ ± 0.62％,52.22％ ± 0.72％,42.52％ ± 0.90％,45.96％ ± 2.11％,29.96％ ± 0.70％ respectively,F =272.0,P ＜ 0.05).Moreover,all the tea polyphenol groups showed significantly increased cell apoptosis rate compared with the control group (all P ＜ 0.05).Fluorescence microscopy showed karyopyknosis,nuclear fragmentation and other typical apoptotic morphological changes in H8 cells in tea polyphenols groups.There were significant differences in the percentage of cells in G1,G2 phase and cell proliferation index among the 5 groups (all P ＜ 0.05).Compared with the control group,the 6.25-,12.5-,25-mg/L tea polyphenols groups showed significantly increased percentage of cells in G1 phase (55.96％ ± 0.72％,54.12％ ± 3.20％,65.30％ ± 1.51％ respectively,all P ＜ 0.05),but significantly decreased percentage of cells in G2 phase (3.17 ± 1.82％,4.94 ± 1.46％,4.65 ± 4.26％ respectively,all P ＜ 0.05) and lower cell proliferation index(0.44 ± 0.01,0.46 ± 0.02,0.36 ± 0.01 respectively,all P ＜ 0.05).Conclusion Tea polyphenols can inhibit the proliferation of H8 cells,induce cell apoptosis,and block cell cycle progression.

Objective To investigate the expression and prognostic value of serum antimicrobial peptide LL37 and human soluble scavenger receptor cysteine-rich domain-containing protein(SSC5D)in elderly patients with chronic heart failure(CHF).Methods A total of 100 CHF patients admitted to Suzhou Kowloon Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2017 to January 2022 were selected as the CHF group.According to the prognosis,these patients were divided into good group(n=60)and poor prognosis group(n=40).Another 100 healthy examinees were selected as the control group.Enzyme-linked immunosorbent assay was used to detect serum levels of LL37 and SSC5D.The differences in serum LL37 and SSC5D levels between CHF group and control group were compared.Spearman correlation analysis was used to analyze the correlation between serum LL37 and SSC5D.Kaplan-Meier survival curve was used to analyze the relationship between serum LL37 and SSC5D expression and the prognosis of CHF patients.COX regression analysis was used to analyze the factors affecting the prognosis of CHF patients.ROC curve was used to analyze the predictive value of serum LL37 and SSC5D for poor prognosis in elderly CHF patients.Result The levels of serum LL37(771.38±158.25 ng/ml)and SSC5D(15 789.35±1 306.25 pg/ml)in CHF group were higher than those in control group(526.23±115.58 ng/ml,8 938.72±858.29 pg/ml),and the differences were significant(t=12.510,43.830,all P＜0.001).Spearman correlation analysis showed that there was a significant positive correlation between serum LL37 and SSC5D in CHF patients(r=0.629,P＜0.001).The serum LL37 and SSC5D levels in CHF patients were positively correlated with NYHA cardiac function classification(r=0.776,0.751,all P＜0.001).The survival rate of the high-level LL37 group was lower than that of the low-level LL37 group(38.18％vs 86.67％),and the difference was significant(Log rankx2=24.242,P＜0.001).The survival rate of the high level SSC5D group was lower than that of the low level SSC5D group(37.74％vs 85.10％),and the difference was significant(Log rank x2=23.291,P＜0.001).Compared with the good prognosis group,the poor prognosis group had a higher proportion of patients over 70 years old,proportion of patients with NYHA cardiac function class Ⅲ+Ⅳ,and serum LL37 and SSC5D levels,and the differences were significant(x2=10.774,4.118,t=4.723,14.059,all P＜0.05).Multivariate COX analysis showed that age＞70 years(OR=1.515,95％CI:1.224～1.858),NYHA cardiac function class Ⅲ+Ⅳ(OR=1.236,95％CI:1.198～1.963),high level of LL37(OR=1.705,95％CI:1.163～2.582)and high level of SSC5D(OR=1.591,95％C1:1.052～1.916)were independent risk factors for the prognosis of CHF patients.ROC curve analysis showed that the area under the curve of serum LL37 combined with SSC5D for predicting poor prognosis of CHF in the elderly was larger than that of serum LL37 and SSC5D alone,and the differences were significant(Z=2.834,2.168,P=0.005,0.030).Conclusion The serum LL37 and SSC5D levels are increased in patients with CHF,and both are risk factors for poor prognosis in patients with CHF,which can be used as clinical indicators to evaluate the prognosis of CHF.

Objective To evaluate the effects of trimetazidine on cardiac function and autonomic function in patients with coronary heart disease and chronic heart failure. Methods 73 patients with coronary heart disease and chronic heart failure were randomly divided into control group (conventional therapy, n=35) and trimetazidine group (conventional therapy plus oral trimetazidine, n=38). All the patients underwent echocardiography for left ventricular ejection fraction (LVEF) and left ventricular end-diastolic diameter (LVd), and 24-hour Holter electrocardiogram for heart rate turbulence (HRT) indexes, turbulence onset (TO) and turbulence slope (TS) were calculated before and 3 months after treatment. Results 3 months after treatment, LVEF increased, and LVd shrank in both groups (P<0.01). LVEF improved better in the trimetazidine group than in the control group (P<0.001). TO decreased and TS increased in the trimetazidine group (P<0.05). Only TO decreased in the trimetazidine group (P<0.05). TO and TS improved better in the trimetazidine group than in the control group (P<0.05). Conclusion Trimetazidine could improve cardiac function and autonomic function in patients with coronary heart disease and chronic heart failure.