Objective: To investigate the perinatal mortality and its causes in Lishui City, Zhejiang Province, so as to provide insights into the reduction of perinatal mortality. Methods: The perinatal mortality data were collected from three designated monitoring counties (districts) in Lishui City from 2015 to 2020. The changing trend in perinatal mortality and causes of death were analyzed, and the differences of perinatal causes of death and maternal conditions in urban and rural areas were compared. Results: There were 406 perinatal deaths in three monitoring counties (districts) of Lishui City from 2015 to 2020. The perinatal mortality showed a downward trend (χ2trend=5.078, P=0.024), and the average perinatal mortality rate was 5.93‰. The top five causes of perinatal mortality were birth defects, unknown causes, umbilical cord factors, maternal factors and preterm delivery. There was significant difference in the causes of perinatal death between urban and rural areas (χ2=25.574, P<0.001); birth defects, umbilical cord factors and unknown causes were predominant in urban areas, accounting for 75.00% (138 deaths); while birth defects, unknown causes and maternal factors were predominant in rural areas, accounting for 67.57% (150 deaths). The proportion of junior high school education and below was significantly higher in rural women than in urban women with perinatal death (54.50% vs. 26.63%; χ2=32.117, P<0.001), and the proportion of more than 5 antenatal examinations was significantly lower in rural women than in urban women with perinatal death (46.85% vs. 59.24%; χ2=6.195, P=0.012). Conclusions The perinatal mortality appeared a tendency towards a decline in Lishui City from 2015 to 2020, and birth defect was the main cause. Prenatal and postnatal care and tertiary prevention should be strengthened.

Background: Fenbendazole, a dewormer drug, is used widely in the clinical treatment of parasite infections in animals. Recent studies have shown that fenbendazole has substantial effects on tumor growth, immune responses, and inflammatory responses, suggesting that fenbendazole is a pluripotent drug. Nevertheless, the antiviral effects have not been reported.Fenbendazole can disrupt microtubules, which are essential for multiple viruses infections, suggesting that fenbendazole might have antiviral effects. Objectives: This study examined whether fenbendazole could inhibit bovine herpesvirus 1 (BoHV-1) productive infection in cell cultures. Methods: The effects of fenbendazole on viral production, transcription of the immediate early (IE) genes, viron-associated protein expression, and the cellular signaling PLC-γ1/Akt pathway were assessed using distinct methods. Results: Fenbendazole could inhibit BoHV-1 productive infections significantly in MDBK cells in a dose-dependent manner. A time-of-addition assay indicated that fenbendazole affected both the early and late stages in the virus replication cycles. The transcription of IE genes, including BoHV-1 infected cell protein 0 (bICP0), bICP4, and bICP22, as well as the synthesis of viron-associated proteins, were disrupted differentially by the fenbendazole treatment. The treatment did not affect the cellular signaling pathway of PLC-γ1/Akt, a known cascade playing important roles in virus infection. Conclusions Overall, fenbendazole has antiviral effects on BoHV-1 replication.

Objective To evaluate the scavenging effect of crude polysaccharides extracted from Lycium barbarum (LBP) on reactive oxygen species in ultraviolet radiation-induced HaCaT cells,and to explore its possible mechanism.Methods Cultured immortalized human keratiuocyte HaCaT cells were divided into 6 groups:blank control group receiving no treatment,LBP group treated with crude LBP alone,ultraviolet A (UVA) group treated with UVA radiation alone,ultraviolet B (UVB) group treated with UVB radiation alone,UVA + LBP group treated with crude LBP for 24 hours followed by UVA radiation,and UVB + LBP group treated with crude LBP for 24 hours followed by UVB radiation.MTT colorimetry was performed to evaluate the cellular proliferative activity,UV spectrophotometric method to measure the UVA and UVB absorption of crude LBP,dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe assay to detect the level of ROS,enzymatic-biochemical method to estimate the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px),as well as to detect the leakage of lactate dehydrogenase (LDH).Results Crude LBP at different concentrations of 0,100,200,300,400,500,600,1 500,2 000 mg/L had no obvious effects on the proliferative activity of HaCaT cells.Crude LBP had a high transmittance of ultraviolet rays at 280-400 nm.Compared with the blank control group,the UVA group and UVB group both showed significantly higher LDH leakage and ROS level,lower activities of SOD and GSH-Px (P ＜ 0.001 or 0.05).Pretreatment with crude LBP before the ultraviolet radiation could significantly increase the activities of SOD and GSH-Px,decrease the LDH leakage and ROS level in the UVA + LBP group and UVB + LBP group compared with the UVA group or UVB group (P ＜ 0.05).Conclusion Crude LBP have no effect of sunscreening agents,but can effectively scavenge ROS,decrease LDH leakage,inhibit ultraviolet radiation-induced photodamage in HaCaT cells,which may be associated with the enhancement of antioxidant enzyme activity.

Objective:To investigate the effect of pterostilbene on the growth, apoptosis and autophagy of a human papillomavirus type 16 (HPV-16) -immortalized cervical epithelial cell line H8.Methods:H8 cells were treated with pterostilbene at different concentrations of 0 (control group) , 25, 50, 75, 100 μmol/L for 24 and 48 hours. Cell counting kit-8 (CCK8) assay was performed to evaluate the cellular proliferative activity, flow cytometry was conducted to detect apoptosis and cell cycle, monodansylcadaverine (MDC) staining and fluorescence microscopy were performed to detect autophagy, and Western blot analysis was conducted to determine the expression of the cell cycle-related protein cyclinD1, apoptosis-related proteins caspase-3 and caspase-9, autophagy-related proteins Beclin1, microtubule-associated protein 1 light chain 3 (LC3) -Ⅱ/Ⅰ, ATG5 and P62, as well as HPV oncoproteins E6 and E7. Statistical analysis was carried out by using one-way analysis of variance, repeated measures analysis of variance and least significant difference- t test. Results:After 48-hour treatment with pterostilbene at different concentrations of 0, 25, 50, 75, 100 μmol/L, the relative cellular proliferation rate significantly differed among the groups (100.00% ± 1.56%, 99.02% ± 4.97%, 93.59% ± 2.01%, 81.28% ± 4.90%, 69.17% ± 7.56%, respectively; F = 77.22, P < 0.05) , and gradually decreased along with the increase in the concentration of pterostilbene; compared with the control group, the pterostilbene groups all showed significantly decreased cellular proliferation rate (all P < 0.05) . After 24-hour treatment with pterostilbene, the proportions of H8 cells at G1, G2 and S phases significantly differed among the above groups ( F = 7 845.00, 51.14, 266.50, respectively, all P < 0.05) ; compared with the control group, the pterostilbene groups showed significantly increased proportions of H8 cells at G1 and G2 phases (all P < 0.05) , but significantly decreased proportions of H8 cells at S phase ( P < 0.05) . After 48-hour treatment with pterostilbene, the apoptosis rate was significantly higher in the 25-, 50-, 75- and 100-μmol/L pterostilbene groups (14.66% ± 0.22%, 13.50% ± 0.49%, 14.56% ± 0.19%, 15.30% ± 0.76%, respectively) than in the control group (11.58% ± 0.50%, all P < 0.05) . After 24-hour treatment with pterostilbene, MDC staining showed only a small number of H8 cells with bright dot-like fluorescence in the control group, but increased number of autophagosome-positive H8 cells with bright dot-like fluorescence in the pterostilbene groups. Western blot analysis revealed that there were significant differences in the protein expression of cyclin D1, caspase-3, caspase-9, Beclin1, LC3-Ⅱ/Ⅰ, ATG5, P62, E6 and E7 among the control and pterostilbene groups after 24- and 48-hour treatment with pterostilbene (all P < 0.05) . The treatment with pterostilbene could down-regulate the expression of cyclin D1, E6 and E7, and up-regulate the expression of caspase-3, caspase-9, Beclin1, LC3-Ⅱ/Ⅰ, ATG5 and P62, with significant differences between the control group and most pterostilbene groups in expression of the above proteins (all P < 0.05) . Conclusion:Pterostilbene can inhibit the proliferation of H8 cells, promote their apoptosis and autophagy, and down-regulate the expression of oncogenes E6 and E7.

Objective To investigate the protective effect of lycium barbarum polysaccharide (LBP) on DNA damage of HSF cells induced by UV.Methods We established the model of UV induced photo damage in HSF cells.We detected the viability of HSF cells by using MTT colorimetry.The UV absorption spectrum of LBP was also measured by UV spectrophotometer.The level of ROS was detected by DCFH-DA fluorescent probe method.Comet assay was employed to evaluate the DNA strand breakage damage.Results When the concentration of LBP was less than or equal to 300μg/ml,there was no significant effect on the proliferation of HSF cells (P＞0.05).When the concentration was more than 300 μg/ml,it could inhibit the cell proliferative activities (P＜0.05).Compared to the UV groups,UV+LBP groups can respectively improve the cell proliferation activity (P＜0.05).The absorbance was slight range 280 from 400 nm.Compared with the UV group,the relative fluorescence intensity and the migration distance of UV+ LBP groups were significantly decreased (P＜0.05).Conclusions Lycium barbarum polysaccharide can effectively inhibit the proliferation activity and protect the breakage of DNA strand induced by UV,which is probably due to its action of removing free radicals.

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 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 explore the delayed discount and personality characteristics of impulsive and premeditated male prisoners and their correlation.Methods A total of 120 male prisoners with aggressive behaviors were tested by Self-made General Situation Questionnaire,Impulsive / Premeditated Aggression Scale (IPAS),NEO Five-Factor Inventory (NEO-FFI),Delay Discount Task (DDT).The effective participants were ninety,including 51 in impulsive group and 39 in premeditated group.Analyzed the difference of delayed discount and personality characteristics between the two group.Results (1) In the delayed discount task,the delay discount rat (K) of the impulsive violence group (0.0278 ± 0.0555) was significantly higher than that of the premeditated violence group (0.0042±0.0078).The difference was statistically significant (t=3.004,P =0.004).(2) The scores of agreeableness (35.84 ± 4.08) and conscientiousness (37.04±4.83) in impulsive violence group were lower than that in premeditated violence group(38.46±5.53,42.31±5.96),and the difference was statistically significant (t=-2.587,P=0.011;t =-4.634,P＜0.01).(3) Correlation analysis showed that K value was not significantly correlated with all dimensions of the big five personality(P＞0.05).Conclusion Male impulsive prisoners have faster tendency to delay discounting compared with male premeditated prisoners.Lower agreeableness and conscientiousness are the personality characteristics of male impulsive prisoners.