Objective To explore the mechanism of Lacticaseibacillus paracasei E6 for improving vinorelbine-induced immunosuppression in zebrafish.Methods The intestinal colonization of L.paracasei E6 labeled by fluorescein isothiocyanate(FITC)in zebrafish was observed under fluorescence microscope.In a zebrafish model of vinorelbine-induced immunosuppression,the immunomodulatory activity of L.paracasei E6 was assessed by analyzing macrophage and neutrophil counts in the caudal hematopoietic tissue(CHT),the number of T-lymphocyte,and the expressions of interleukin-12(IL-12)and interferon-γ(IFN-γ).The contents of short-chain fatty acids(SCFAs)in L.paracasei E6 fermentation supernatant and the metabolites of L.paracasei E6 in zebrafish were detected by LC-MS/MS-based targeted metabolomics.The immunomodulatory effects of the SCFAs including sodium acetate,sodium propionate and sodium butyrate were evaluated in the zebrafish model of immunosuppression.Results After inoculation,green fluorescence of FITC-labeled L.paracasei E6 was clearly observed in the intestinal ball,midgut and posterior gut regions of zebrafish.In the immunocompromised zebrafish model,L.paracasei E6 significantly alleviated the reduction of macrophage and neutrophil counts in the CHT,increased the fluorescence intensity of T-lymphocytes,and promoted the expressions of IL-12 and IFN-γ.Compared with MRS medium,L.paracasei E6 fermentation supernatant showed significantly higher levels of acetic acid,propionic acid and butyric acid,which were also detected in immunocompromised zebrafish following treatment with L.paracasei E6.Treatment of the zebrafish model with sodium acetate and sodium propionate significantly increased macrophage and neutrophil counts in the CHT and effectively inhibited vinorelbine-induced reduction of thymus T cells.Conclusion L.paracasei E6 can improve vinorelbine-induced immunosuppression in zebrafish through its SCFA metabolites acetic acid and propionic acid.

Objective To investigate the efficacy of Lactobacillus plantarum ZG03(L.plantarum ZG03)for ameliorating oxidative stress in zebrafish.Methods We evaluated the growth pattern of L.plantarum ZG03,observed its morphology using field emission scanning electron microscopy,and assessed its safety and potential efficacy with whole-genome sequencing for genetic analysis.FITC-labeled ZG03 was used to observe its intestinal colonization in zebrafish.In a zebrafish model of 2%glucose-induced oxidative stress,the effect of ZG03 was evaluated by assessing the changes in neutrophils in the caudal hematopoietic tissue(CHT),superoxide dismutase(SOD)activity,reactive oxygen species(ROS)levels,and malondialdehyde(MDA)content.Liquid chromatography-mass spectrometry-based targeted metabolomics was used for analyzing short-chain fatty acids(SCFAs)in the zebrafish,and the antioxidant effects of the key metabolites(acetate,propionate,and caproate)were tested.Results On MRS agar,L.plantarum ZG03 formed circular,smooth,moist,and milky-white colonies with a rod-shaped cell morphology.Genomic analysis revealed abundant sugar metabolism gene clusters.After inoculation of FITC-labeled L.plantarum ZG03 in zebrafish,green fluorescence was clearly observed in the intestinal bulb,mid-intestine,and hind intestine.In zebrafish with glucose-induced oxidative stress,L.plantarum ZG03 significantly reduced ROS levels and the number of neutrophils in the CHT with increased SOD activity.L.plantarum ZG03 significantly increased the content of SCFAs including acetic acid,propionic acid,and caproic acid in zebrafish metabolites.In addition,sodium acetate,sodium propionate,and sodium caproate in the SCFAs significantly increased SOD activity in the zebrafish models.Conclusion L.plantarum ZG03 ameliorates oxidative stress in a glucose-induced zebrafish model through its metabolites,particularly the SCFAs including acetic acid,propionic acid and caproic acid.

AIM To investigate the clinical effects of Mahuang Fuzi Xixin Decoction combined with non-immunosuppressive treatment on patients with focal segmental glomerulosclerosis due to Spleen-Kidney Yang Deficiency.METHODS Sixty patients were randomly assigned into control group(30 cases)for 24-week intervention of non-immunosuppressive treatment,and observation group(30 cases)for 24-week intervention of both Mahuang Fuzi Xixin Decoction and non-immunosuppressive treatment.The changes in clinical effects,TCM syndrome effects,TCM syndrome score,24 h UTP,ALB,eGFR,Nephrin,Podocin and safety indices were detected.RESULTS The observation group demonstrated higher total effective rate and TCM syndrome effective rate than the control group(P＜0.05).After the treatment,the two groups displayed decreased TCM syndrome score,24 h UTP,Nephrin,Podocin(P＜0.05),and increased ALB(P＜0.05),especially for the observation group(P＜0.05).No obvious adverse reactions occured in the two groups.CONCLUSION For the patients with focal segmental glomerulosclerosis due to Spleen-Kidney Yang Deficiency,Mahuang Fuzi Xixin Decoction combined with non-immunosuppressive treatment can improve TCM syndromes,whose action mechanism may be contribute to the alleviation of podocyte injury.

AIM To optimize the extraction process for total polyphenols from Conioselinum vaginatu(Spreng.)Thell.,make component analysis,and evaluate their anti-oxidant,hypoglycemic activities.METHODS The effects of ultrasound,enzymatic hydrolysis,acid hydrolysis,alcohol extraction and hydrolysis processes on the extraction quantity of total polyphenols were investigated,respectively.With extraction temperature,extraction time,ethanol concetration and liquid-solid ratio as influencing factors,extraction quantity of total polyphenols as an evaluation index,the extraction process was optimized by response surface method.HPLC was adopted in the identification of polyphenolic composition and determination of their contents.Subsequently,total polyphenols' scavenging capacities on DPPH,ABTS,OH free radicals,total reducing power and inhibitory capacity on α-glucosidase were determined.RESULTS The highest extraction quantity of total polyphenols was observable when extraction process was employed.The optimal conditions were determined to be 62 ℃ for extraction temperature,54 min for extraction time,69％for ethanol concentration,and 50∶1 for liquid-solid ratio,the extraction quantity of total polyphenols was(9.51±0.2)mg GAE/g.Seven constituents existed in C.vaginatu,among which ferulic acid demonstrated the highest content,followed by that of myricetin,while D-tryptophan content was the lowest.At the concentration of 7.61 mg/L,total polyphenols displayed the scavenging rates on DPPH,ABTS,OH free radicals of 80.70％,85.97％,28.60％,total reducing power of 0.22,and inhibition rate on α-glucosidase of 77.23％,respectively.CONCLUSION This stable and reliable method can be used for the extraction of total polyphenols from C.vaginatum with strong anti-oxidant,hypoglycemic activities.

Objective To evaluate the safety and efficacy of valve-in-valve transcatheter mitral valve replacement(ViV-TMVR)in patients with bioprosthetic valve degeneration who are at high surgical risk.Methods This study is a multi-center,retrospective cohort analysis of 20 consecutive patients who underwent transseptal ViV-TMVR using the Edwards SAPIEN 3 transcatheter heart valve(THV).The primary endpoints include technical success and procedural success,both defined according to the Mitral Valve Academic Research Consortium(MVARC)criteria,as well as mortality and functional change assessed based on New York Heart Association(NYHA)classification at 30-days and six months post-procedure.Clinical follow-up assessments are conducted at 30-days and six months.Results From February 2021 to October 2022,a total of 20 patients with symptoms of bioprosthetic valve degeneration were enrolled across nine sites in China.The patients had a mean age of(73.5±5.5)years,with 85.0％being females and 70.0％classified as NYHA class Ⅲ/Ⅳ.The study achieved a 100.0％technical success rate and a 90.0％procedural success rate finally.All patients remained alive during the 30-day follow-up period.However,six months post-intervention,two patients(10.0％)were re-hospitalized due to heart failure,and sadly,one of them(5.0％)died.None of the patients reported any adverse events related to ViV-TMVR during the follow-up period.Notably,there was a significant improvement in NYHA class compared to baseline(P=0.0004)at six-month follow-ups.Conclusions The transseptal ViV-TMVR technique proved to be highly successful and was associated with significant improvement in NYHA class function.These findings strongly suggest that it serves as a safe and efficient treatment alternative for high-risk patients suffering from bioprosthetic valve degeneration.

Aim To explore the mechanism of Jiawei Shaofu Zhuyu decoction in antagonizing endometriosis fibrosis by regulating mitophagy.Methods After the animal model was constructed,the syndrome was evalu-ated by general condition,organ water content and ther-mal imaging.The curative effect was evaluated by the weight of ectopic focus and the degree of adhesion.The pathological changes were compared using HE stai-ning,transmission electron microscopy,Masson and Sir-ius red staining.The expression of PINK1 and Parkin was detected by immunohistochemistry.The expression of mRNA and protein was determined by qPCR and Western blot,and the level of serum ROS was detected by ELISA.Results The autonomic activity of model mice was weakened,the water content of organs rose,and the temperature of limbs and lower abdomen was reduced by thermal imaging.HE staining showed obvi-ous hyperplasia of ectopic epithelium and glands.Transmission electron microscopy showed mitochondrial and endoplasmic reticulum structure damage,and nor-mal autophagy structure disappeared.Masson and Siri-us red staining showed increased collagen deposition;immunohistochemistry showed decreased expression of PINK1 and Parkin in ectopic foci.qPCR and Western blot showed that the expression of PINK1,Parkin,Bec-lin1,LC3 mRNA and protein in ectopic foci of model mice decreased,the expression of p62 mRNA and pro-tein increased,and serum ROS increased.The syn-drome performance of model mice was improved after the intervention of Jiawei Shaofu Zhuyu decoction;the inflammatory infiltration of ectopic foci was relieved,the morphology of mitochondria and endoplasmic retic-ulum was restored,and normal autophagy structure ap-peared.The degree of collagen deposition and fibrosis was reduced;the mRNA and protein expression of PINK1,Parkin,Beclin1 and LC3 increased.The ex-pression of p62 mRNA and protein decreased,and the level of ROS decreased.Conclusions Jiawei Shaofu Zhuyu decoction can improve the fibrosis of ectopic le-sions in mice with endometriosis of cold-dampness sta-sis syndrome,which may be related to the regulation of mitophagy.

Objective:To compare the therapeutic efficacy and safety of local anesthesia and spinal anesthesia for the patients with varicocele(VC)who underwent microsurgical varicocelectomy(MV).Methods:We retrospectively analyzed the data of VC patients who underwent MV treatment at the Andrology Department of the Affiliated Ruikang Hospital of Guangxi University of Chinese Medicine from May 2020 to March 2023.Cases with complete clinical data and follow-up evaluation were selected and divided into a control group(spinal anesthesia)and an observation group(local anesthesia)according to different anesthesia methods.The surgical time(including anesthesia time),visual analogue scale(VAS)score for pain,hospital stay,treatment cost,sperm concentration,for-ward motile sperm rate,and normal sperm morphology rate after three months of surgery,as well as postoperative complications and re-currence rate were compared between the two groups.Results:A total of 107 eligible cases were included,with 56 cases in the con-trol group and 51 cases in the observation group.There was no significant difference in the VAS score for pain during and after four hours of surgery,as well as postoperative complications,and recurrence rate between the two groups(P＞0.05).There was an signif-icant increase in sperm concentration,forward motile sperm rate,and normal sperm morphology rate in both of two groups after three months of surgery(P＜0.05).However,there was no significant difference between the two groups three months after surgery(P＞0.05).The surgical time and hospital stay were shorter than those of the control group(P＜0.05).And the treatment cost in observa-tion group was lower than that of the control group(P＜0.05).Conclusion:Both local anesthesia and lumbar anesthesia for MV treatment of VC have good efficacy and safety.However,patients treated with MV under local anesthesia for VC have obvious advanta-ges in terms of operation time(including anesthesia time),hospital stay,and treatment cost,which is worthy of clinical promotion and application.

Objective To establish a rapid,sensitive,and specific multiplex PCR detection method for the simultaneous detection of Cryptosporidium,Eimeria,and bovine parvovirus.Methods Specific primers targeting the SSU rRNA genes of Cryptosporidium and Eimeria,as well as the VP2 gene of bovine parvovirus were designed and the corresponding recombinant plasmid standards were constructed.To establish the multiplex PCR method,the reaction conditions were optimized using temperature gradient PCR and single-variable control methods.The sensitivity,specificity,reproducibility,and clinical application of the protocol were evaluated.Results The optimal annealing temperature was found to be 60.5℃,and the forward and reverse primer concentrations were determined to be 0.2 μmol/L for Eimeria,and 0.4 μmol/L for Cryptosporidium and bovine parvovirus.The assay demonstrated high sensitivity,with detection limits of 243,260,and 3 110 copies for the recombinant plasmid standards of Cryptosporidium,Eimeria,and bovine parvovirus,respectively.Specificity testing showed no cross-reactivity with ten common bovine pathogens,including Salmonella,bovine viral diarrhea virus,and bovine rotavirus.Consistent intra-and inter-batch results confirmed the strong reproducibility of the method.Clinical application to 81 diarrhea samples from various regions in the Ganzi Prefecture,Sichuan,revealed positivity rates of 18.52%(15/81)for Cryptosporidium,34.57%(28/81)for Eimeria,and 18.52%(15/81)forbovineparvovirus,withamixedinfectionrateof3.7%(3/81).Conclusions Themultiplex PCR method established in this study offers a reliable tool for differential diagnosis and epidemiological investigation of the three common diarrheal pathogens in yaks.

This study was aimed at determining the molecular characteristics of Yersinia pestis in the natural plague foci around Qinghai Lake through single nucleotide polymorphism technology,to lay a foundation for molecular epidemiological and source-tracing analysis of Y.pestis in this area.Using the whole genome sequencing technology,we obtained the whole genome sequences of 84 representative Y.pestis strains.Using the sequences of Y.pestis and Yersinia pseudotuberculosis IP32953 from the NCBI database as references,we compared and analyzed the 2 298 SNP loci of these strains.From 1957 to 2020,84 representative strains of Y.pestis from the natural plague foci around Qinghai Lake were divided into two clades:1.IN2 and 3.ANT1.The 1.IN2 clade was the characteristic population of Y.pestis throughout all epidemic years in this area.Additionally,analysis of the SNP distribution and hosts in the region indicated that the 1.IN2 clade was located in five counties except Wulan,whereas the 3.ANT1 clade was isolated from Himalayan marmot and dog in two counties.In conclusion,the population structure of SNP of Y.pestis in the natural plague foci around Qinghai Lake is relatively simple,and SNP analysis of Y.pestis provided a scientific basis for tracing plague epidemic sources and formulating plague prevention and control measures in this area.

Objective:To explore the incidence status,influencing factors and short-term prognosis characteristics of early onset coronary heart disease in women in Wansheng District of Chongqing,and to provide scientific basis for formulating regional prevention and treatment strategies.Methods:This study was a single-center retrospective study,100 coronary heart disease in women from January 2022 to December 2023 at Chongqing Wansheng Economic and Technological Development Zone People's Hospital were prospective selected,and they were divided into early onset group of 40 cases(≤ 65 years old)and late onset group of 60 cases(＞65 years old)based on their age of onset.Another 60 healthy women who underwent physical examinations during the same period to exclude coronary heart disease were selected as the control group.Univariate factor and multiple factor logistic regression analysis were used to identify independent risk factors for early onset coronary heart disease in women.Draw receiver operating characteristic(ROC)curve for the subjects,the efficacy of risk factors in predicting early onset coronary heart disease based on the area under the curve(AUC)of ROC curve were evaluated.Patients were followed up for 1 year to observe the occurrence of major adverse cardiovascular events(MACE).Result:Among 100 fcoronary heart disease in women,the early onset group accounted for 40.00％(40/100).Univariate analysis showed that age,hyperlipidemia history,smoking history,hypertension history,family history,diabetes history,total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C)were related to the early onset coronary heart disease.Multivariate analysis showed that,hyperlipidemia history(OR=4.124,95％CI:2.343-7.217),smoking history(OR=3.564),hypertension(OR=3.253),family history(OR=2.981),diabetes history(OR=2.873)were independent risk factors.ROC curve analysis results showed that joint evaluation had the best predictive value,with AUC of 0.829,which was higher than the AUC of individual evaluation for each factor.The incidence of MACE in the early onset group(45.00％)was significantly higher than that in the late onset group(P＜0.05).Conclusion:Early onset coronary heart disease in women in Wansheng District of Chongqing is related to the hyperlipidemia history,smoking,hypertension history,family history and diabetes history.The incidence of MACE in early-onset patients followed up for 1 year is higher than that in late-onset patients.