ObjectiveTo explore the mechanism of Xianfang Huomingyin （XFHMY） in the treatment of type Ⅲ prostatitis （CP/CPPS） through network pharmacology， molecular docking， and animal experiments. MethodsThe traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Swiss Target Prediction database were used to screen and sort out the active ingredients and corresponding targets of XFHMY. The potential therapeutic targets of CP/CPPS were collected from online databases， such as the Online Mendelian Inheritance in Man （OMIM）， GeneCards， and DisGeNET. The potential core targets of XFHMY for treating CP/CPPS were further screened by constructing a protein-protein interaction （PPI） network and performing topological analysis. Meanwhile， the DAVID database was chosen to perform enrichment analysis on the intersection targets. On this basis， the AutoDock software was used for molecular docking， and the data was subsequently imported into the GraphPad Prism 8 software to generate a heat map. SD rats were divided into seven groups： A blank group， a sham operation group， a model group， low-， medium-， and high-dose XFHMY groups （3.645， 7.29， 14.58 g·kg^-1）， and a tamsulosin hydrochloride group （0.018 mg·kg^-1）. Hematoxylin-eosin （HE） staining was used to evaluate the pathological changes in prostate tissue. The inflammatory factor indicators of rats in each group were detected via enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative reverse transcription polymerase chain reaction （Real-time PCR） and Western blot were used to evaluate the mRNA and protein expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， and nuclear transcription factor-κB （NF-κB） p65 in prostate tissue. ResultsThe HE staining showed no significant signs of inflammatory cell infiltration in the prostate of the sham operation group compared to the blank group， while the model group had significantly inflammatory cell infiltration. The ELISA results showed that compared to the blank group， TNF-α， IL-1β， and COX-2 in the sham operation group had no significant differences. However， they were significantly higher in the model group （P<0.01）， indicating successful CP/CPPS modeling in rats. Compared with the model group， the low-，medium-and high-dose XFHMY group and the tamsulosin hydrochloride group showed significant decreases in TNF-α， IL-1β， and COX-2 （P<0.05，P<0.01）. The Real-time PCR analysis revealed that compared to the model group， the low-dose XFHMY group had reduced Akt and NF-κB p65 mRNA expression（P<0.05，P<0.01）. In the medium-and high-dose XFHMY group and tamsulosin hydrochloride group， PI3K， Akt， and NF-κB p65 mRNA levels decreased significantly（P<0.05，P<0.01）. Western blot analysis showed that compared to the model group， the low-dose XFHMY group had lower p-NF-κB p65/NF-κB p65 （P<0.05）. The medium- and high-dose XFHMY group and the tamsulosin hydrochloride group showed significant decreases in p-PI3K/PI3K， p-Akt-ser473/Akt， p-Akt-thr308/Akt， and p-NF-κB p65/NF-κB p65 （P<0.01）. ConclusionXFHMY may exert therapeutic efficacy on CP/CPPS by inhibiting the PI3K/Akt/NF-κB signaling pathway and reducing inflammatory responses. Additionally， NF-κB activation may be related to the activation of ser473 and thr308 sites.

ObjectiveTo observe the effects of Yangjing Zhongyutang （YJZYT） on mitochondrial biogenesis and oxidative stress damage mediated by the silent information regulator 1 （SIRT1）/peroxisome proliferator-activated receptor gamma coactivator-1alpha （PGC-1α） signaling pathway in cyclophosphamide （CTX）-induced rats with diminished ovarian reserve （DOR）， and to explore its mechanism in improving ovarian reserve function and follicular development. MethodsForty-two 8-week-old female SD rats with normal estrous cycles were randomly divided into a blank control group （n=7） and a model group （n=35）. Rats in the model group received a single intraperitoneal injection of CTX （90 mg·kg^-1） to establish the DOR model. After modeling， estrous cycles were monitored for 7 consecutive days， and model success was confirmed based on criteria for estrous cycle disruption. After successful modeling， rats were divided into groups for intervention： estradiol valerate group （0.09 mg·kg^-1）， and YJZYT high-， medium-， and low-dose groups （19.98， 9.99， 5.00 g·kg^-1）. The blank control group and model group were given an equal volume of distilled water by gavage. All groups received daily gavage once for 4 consecutive weeks. The general state， body weight， and ovarian wet weight of rats were observed and recorded， and the ovarian organ index was calculated. Enzyme-linked immunosorbent assay （ELISA） was used to measure serum levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E₂）， anti-Müllerian hormone （AMH）， superoxide dismutase （SOD）， and glutathione peroxidase （GSH-Px）. Hematoxylin-eosin （HE） staining was performed to observe ovarian histomorphological changes and follicular development status. Immunofluorescence was used to detect reactive oxygen species （ROS） expression levels. Colorimetric assays were employed to measure adenosine triphosphate （ATP） and malondialdehyde （MDA） content in ovarian tissues. Quantitative Real-time polymerase chain reaction （Real-time PCR） was used to detect mitochondrial DNA （mtDNA） copy number and the mRNA expression levels of key genes including SIRT1， PGC-1α， nuclear respiratory factor 1 （NRF1）， and mitochondrial transcription factor A （TFAM）. Western blot was performed to detect the protein expression levels of SIRT1， PGC-1α， NRF1， and TFAM. ResultsCompared with the blank group， rats in the model group exhibited disrupted estrous cycles， obviously reduced body weight， and decreased ovarian index （P<0.05）. Ovarian histopathology revealed cortical thinning， loose structure， and a significant reduction in both primordial and growing follicles （P<0.01）. Serum FSH and LH levels were significantly elevated （P<0.01）， while E₂ and AMH levels were obviously reduced （P<0.05， P<0.01）. ATP content and mtDNA copy number decreased in ovarian tissue （P<0.01）， ROS expression increased， MDA levels rose， while SOD and GSH-Px activities obviously decreased （P<0.05， P<0.01）， mRNA and protein expression levels of SIRT1， PGC-1α， NRF1， and TFAM were obviously downregulated （P<0.05， P<0.01）. After treatment， compared with the model group， body weight and ovarian index obviously recovered in rats administered various doses of YJZYT （P<0.05）， serum E₂ and AMH levels increased， while FSH and LH levels obviously decreased （P<0.05， P<0.01）， ovarian tissue ATP content and mtDNA copy number were up-regulated， ROS and MDA levels decreased， and antioxidant enzymes SOD and GSH-Px activity obviously increased （P<0.05， P<0.01）， Gene and protein expression levels related to the SIRT1/PGC-1α /NRF1/TFAM signaling pathway were obviously up-regulated compared to the model group （P<0.05， P<0.01）， HE staining revealed that ovarian structure gradually recovered to integrity in all treatment groups， with a obviously increase in the number of primordial and growing follicles （P<0.05， P<0.01）. Granulosa cells were neatly arranged， indicating marked improvement in ovarian function. ConclusionYJZYT may improve ovarian function and follicular development in rats with diminished ovarian reserve by activating the SIRT1/PGC-1α signaling pathway， promoting mitochondrial biogenesis， enhancing mitochondrial function， and alleviating oxidative stress damage.

Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.

Objective:To assess the association between diabetes mellitus and the risk of sudden cardiac death (SCD), and to identify potential contributing factors.Methods:This meta-analysis was an updated version of the original study Diabetes mellitus and the risk of sudden cardiac death： a systematic review and meta-analysis of prospective studies. The original review included all eligible case-control and cohort studies published in PubMed and Embase up to 2017 that investigated the association between diabetes and SCD risk. In this updated study, newly published studies were added, including those available in PubMed, Embase, China National Knowledge Infrastructure (CNKI), and WANFANG MED ONLINE up to December 3, 2023. Search terms included "diabetes""glucose""sudden cardiac death" "cardiac arrest" and their Chinese equivalent. The primary outcome was the risk of SCD, while factors such as country, ethnicity, skin color, follow-up duration, left ventricular ejection fraction (LVEF), baseline comorbidities, and other relevant variables were analyzed as potential influencing factors. Relative risk ( RR) was used as the summary measure. A random-effects model was used when significant heterogeneity was detected, otherwise a fixed-effects model was used. Cochran′s Q test was used for subgroup analysis to assess the influence of factors such as region, baseline diseases, LVEF, and ethnicity (based on skin color) on the outcomes. Results:A total of 32 cohort/case-control studies with a combined sample size of 3 252 954 individuals were included. The meta-analysis showed that the risk of SCD in patients with diabetes was double that of non-diabetics ( RR=2.00, 95% CI: 1.83-2.19, P<0.001). In Asian populations, the risk of SCD in diabetic patients was 1.78 times that of non-diabetic individuals ( RR=1.78, 95% CI: 1.51-2.10), 2.05 times that of in European populations ( RR=2.05, 95% CI: 1.79-2.34), and 2.12 times that of in American populations ( RR=2.12, 95% CI: 1.82-2.47), with no statistically significant heterogeneity between regions ( P=0.287). Among individuals without other baseline comorbidities, the risk of SCD was 2.12 times higher in diabetic patients than in those without diabetes ( RR=2.12, 95% CI: 1.89-2.38). In patients with baseline coronary heart disease, the risk was 1.75 times that of non-diabetics ( RR=1.75, 95% CI: 1.45-2.11). In those with baseline heart failure, the risk was 1.92 times that of non-diabetics ( RR=1.92, 95% CI: 1.51-2.43). In patients with baseline atrial fibrillation, the risk was 4.00 times that of non-diabetic individuals ( RR=4.00, 95% CI: 1.38-11.56). In patients undergoing hemodialysis due to renal failure, the risk was 1.76 times that of non-diabetic individuals ( RR=1.76, 95% CI: 1.25-2.48), with no statistically significant heterogeneity between groups ( P=0.262). In cardiac patients with LVEF>50%, the risk of SCD in diabetic patients was 2.08 times that of non-diabetic individuals ( RR=2.08, 95% CI: 1.57-2.75), and in those with LVEF<50%, the risk was 1.69 times that of non-diabetic individuals ( RR=1.69, 95% CI: 1.30-2.18), with no statistically significant heterogeneity between groups ( P=0.277). In yellow-skinned populations, the risk of SCD in diabetic patients was 1.80 times that of healthy individuals ( RR=1.80, 95% CI: 1.73-1.87), and in white-skinned populations, it was 2.18 times that of healthy individuals ( RR=2.18, 95% CI: 1.88-2.54), with statistically significant heterogeneity between groups ( P=0.014). Conclusions:Diabetes mellitus significantly increased the risk of SCD, and this effect may be more pronounced in white-skinned populations, while region, baseline comorbidities, and LVEF had no further effect.

Objective:To investigate the efficacy and safety of TC (paclitaxel and carboplatin) regimen combined with bevacizumab in the treatment of advanced ovarian cancer.Methods:A prospective randomized controlled study was conducted. A total of 102 patients with advanced ovarian cancer admitted to Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University from January 2017 to April 2021 were selected and divided into the observation group and the control group according to random number table method, 51 cases in each group. The observation group was treated with TC regimen combined with bevacizumab, and the control group was treated with TC regimen. The clinical efficacy, carbohydrate antigen 125 (CA125) level, human epididymal protein 4 (HE4) level, thymidine kinase 1 (TK1) level, adverse reactions, the overall survival time, recurrence rate and mortality were compared between the 2 groups.Results:There were no statistically significant differences in baseline data such as age, tumor staging and tumor type between the observation group and the control group (all P > 0.05). The effective rate and the disease control rate of the observation group were 58.82% (30/51) and 88.24% (45/51), respectively, which were higher than those of the control group [37.25% (19/51) and 64.71 (33/51)], and the differences were statistically significant ( χ2 = 4.75, P = 0.029; χ2 = 7.85, P = 0.005). Before treatment, there were no statistically significant differences in CA125, HE4 and TK1 levels between the 2 groups (all P > 0.05). After treatment, CA125, HE4 and TK1 levels in the observation group were lower than those in the control group (all P < 0.05). There were no statistically significant differences in the incidence of myelosuppression, liver function damage, gastrointestinal reaction, proteinuria, diarrhea and abdominal pain, high blood pressure between the observation group and the control group (all P > 0.05). The 2-year overall survival rate of the observation group was higher than that of the control group (82.0% vs. 64.7%), while the recurrence rate of the observation group was lower than that of the control group [21.57% (11/51) vs. 45.10% (23/51)], and the differences were statistically significant (all P < 0.05). Conclusions:TC regimen combined with bevacizumab has favorable therapeutic effects and good safety in the treatment of advanced ovarian cancer, which is beneficial to prolong the survival time of patients.

【Objective】 To investigate the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (M-TESE) in patients with non-obstructive azoospermia (NOA) caused by different causes. 【Methods】 A retrospective analysis was performed on 225 NOA patients during Jan.2020 and Dec.2022. The relation between SRR and patients’ age,body mass index (BMI),testicular volume,endocrine hormones and different etiological classifications were analyzed. 【Results】 According to whether sperm was obtained by surgery,the patients were divided into two groups,including 107 cases in the sperm group and 118 cases in the non-sperm group. There were no significant differences in patients’ age,testicular volume and levels of endocrine hormones between the two groups (P>0.05). According to the different causes,NOA patients with mumps history,cryptorchidism history,AZFc deletion or Klinefelter syndrome (KS) had higher SRR,while idiopathic NOA patients had the lowest SRR (P<0.05). 【Conclusion】 M-TESE is an effective treatment of NOA. There is no correlation between SRR and patients’ age,MBI,testicular volume and levels of endocrine hormones. NOA caused by different etiological classifications may have different SRR.

Objective To study the effect of mitochondrial reactive oxygen species(ROS)on male fertility and its application value in assisted reproduction so as to provide a new method for fertility assessment and etiological treatment of male infertility.Methods The relationship between sperm ROS and male infertility,sperm nuclear DNA integrity and sperm membrane function was analyzed.Then the relationships of the ROS of sperms for fertiliza-tion with fertilization rate,cleavage rate,embryo rate and pregnancy outcome were analyzed.Results(1)The percentage of the high ROS in sperm membrane of the male infertility group was significantly higher than that of the normal fertility group(P<0.01),but there was no statistical difference in the percentage of the high ROS in the sperm mitochondria between the two groups.(2)The normal rate of sperm membrane function in the normal group was significantly higher than that in the abnormal group(P<0.01),but there was no significant difference in the sperm DNA fragmentation index(DFI)between the groups.(3)There was significant difference in the fertilization rate between the IVF control group and IVF observation group,but no significant difference was seen in the cleavage rate,excellent embryo rate and clinical pregnancy rate between the groups.Conclusion Sperm ROS is related to male fertility to some extent.Abnormal ROS may lead to impaired sperm membrane function and affect sperm fertilization ability,thus affecting male fertility.The detection of ROS in sperm can be used as a new method to evaluate male fertility and provide a basis for diagnosis or treatment of male infertility.For patients with ART or infertility,the appropriate time to conceive can be selected according to the sperm ROS level.

To optimize the quantitative detection method for Salmonella enterica and analyze the quantitative contamination level of Salmonella enterica in raw pork samples from farmer′s markets in Chengdu. Based on qualitative detection standard method of Salmonella enterica in China (GB 4789.4-2016) and the quantitative detection method of FSIS in the United States (MLG 4.08 and MLG appendix 2.05 MPN), the selective enrichment broth, screening plate, identification method and quantitative dilution ratio in quantitative detection of Salmonella enterica were optimized using 70 samples of raw pork. The optimized method compared by student′s t-test was used to detect 40 samples of raw pork collected from farmer′s markets in Chengdu from June to October 2020. For isolation of Salmonella from raw pork samples, the coincidence degree of TTB enrichment solution was significantly higher than that of RV enrichment solution (0.93±0.32 vs 0.35±0.62, t=8.324, P=0.001) and the consistency of suspicious colonies screened by XLT4 plate was significantly higher than that of Salmonella chromogenic medium (0.77±0.09 vs 1.00±0.00, t=2.971, P =0.017). The MPN method used 4 successive gradient dilutions, namely 12 tube method, could obtain more accurate quantitative value for Salmonella enterica. The combined use of selective enrichment broth TTB, XLT4 plate, Real-time PCR and MALDI-TOF mass spectrometry could get better results for screening and identifying Salmonella enterica. The detection rate for Salmonella enterica isolated from raw pork in farmer′s markets was 92.5% (37/40). The most of the Salmonella positive samples (83.8%, 31/37) were detected with a contamination level ranged from 0.1 to 55 MPN/g. The optimized quantitative detection method for Salmonella enterica in raw pork in this study can effectively screen the target bacteria and obtain more accurate quantitative value.

To optimize the quantitative detection method for Salmonella enterica and analyze the quantitative contamination level of Salmonella enterica in raw pork samples from farmer′s markets in Chengdu. Based on qualitative detection standard method of Salmonella enterica in China (GB 4789.4-2016) and the quantitative detection method of FSIS in the United States (MLG 4.08 and MLG appendix 2.05 MPN), the selective enrichment broth, screening plate, identification method and quantitative dilution ratio in quantitative detection of Salmonella enterica were optimized using 70 samples of raw pork. The optimized method compared by student′s t-test was used to detect 40 samples of raw pork collected from farmer′s markets in Chengdu from June to October 2020. For isolation of Salmonella from raw pork samples, the coincidence degree of TTB enrichment solution was significantly higher than that of RV enrichment solution (0.93±0.32 vs 0.35±0.62, t=8.324, P=0.001) and the consistency of suspicious colonies screened by XLT4 plate was significantly higher than that of Salmonella chromogenic medium (0.77±0.09 vs 1.00±0.00, t=2.971, P =0.017). The MPN method used 4 successive gradient dilutions, namely 12 tube method, could obtain more accurate quantitative value for Salmonella enterica. The combined use of selective enrichment broth TTB, XLT4 plate, Real-time PCR and MALDI-TOF mass spectrometry could get better results for screening and identifying Salmonella enterica. The detection rate for Salmonella enterica isolated from raw pork in farmer′s markets was 92.5% (37/40). The most of the Salmonella positive samples (83.8%, 31/37) were detected with a contamination level ranged from 0.1 to 55 MPN/g. The optimized quantitative detection method for Salmonella enterica in raw pork in this study can effectively screen the target bacteria and obtain more accurate quantitative value.

Objective To investigate clinical effect of kidney essence granule on DNA intergrity in patients with idiopathic weak sperm.Methods 110 cases of idiopathic weak sperm disease in outpatient service of male infertility were selected, and divided into 3 groups using random number table method, treatment group (n=40) treated with strong kidney essence granule and control group (n=40) by Wuzi Yanzong pill treatment, blank control group (n =30) given placebo treatment.Semen parameters and sperm DNA fragmentation index (DFI) differences were compared within groups. Results The efficacy of treatment group was significantly higher than control group and blank control group, and control group was significantly higher than blank control group, the differences were all statistically significant (P<0.05).There was no significant difference among the 3 groups of patients within semen volume, liquefaction time, a,(a+b),(a+b+c) grade sperm percentage, seminal plasma fructose and acid phosphatase level pre-and post-treatment.Pre-treatment, sperm density, sperm abnormality rate, α-glycosides and DFI were not significantly different among the 3 groups.Post-treatment, sperm density in treatment group was significantly higher than that in control group, and sperm deformity rate and DFI were significantly lower than that in control group, and that in control group was significantly lower than that in blank control group (P <0.05) .Pearson correlation analysis showed that the sperm density was negatively correlated with DFI(r=-0.354), and the sperm deformity rate was positively correlated with DFI(r=0.324).Conclusion The main reason for abnormal DNA integrity are renal dysfunction, and kidney essence granule can be used as an effective medicine to improve the integrity of sperm DNA.