ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

ObjectiveSpinal cord injury (SCI) directly impairs the regulatory function of the autonomic nervous system, induces intestinal dysfunction, and significantly reduces patients’ quality of life. Preclinical studies have shown that electroacupuncture (EA) therapy can regulate the brain-gut axis and is used to treat central nervous system diseases such as major depressive disorder, Alzheimer’s disease and Parkinson’s disease. Recent research has established that fecal microbiota transplantation (FMT) from EA-treated SCI rats restored intestinal motility and colonic morphology. However, it remains unclear whether the regulation of gut microbiota by EA therapy directly contributes to neural repair after SCI. This study aims to explore whether gut microbiota mediates the neuroprotective effect of EA in the treatment of SCI and its possible mechanism. MethodsThe study employed RNA transcriptome analysis of spinal cord tissue to characterize gene expression profiles and to identify key signaling pathways following EA treatment for SCI. Hematoxylin-Eosin (HE) staining and Nissl staining were used to observe the morphological changes in spinal cord tissue. Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) were applied to detect the effects of EA on the expression of proteins related to nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) -dependent pyroptosis. Using 16S rDNA sequencing, the study observed alterations in gut microbiota diversity and community composition in SCI rats. Prior to establishing SCI models, rats were pretreated with an antibiotic cocktail to induce gut dysbiosis, and the effects on intestinal function and spinal cord neural repair were evaluated. FMT was performed to investigate the regulatory effects of post-EA FMT on motor function, general status, liver and spleen indices, and NLRP3-mediated pyroptosis in SCI rats. ResultsEA improved motor function and reduced regulated neuronal cell death in SCI rats. Transcriptomic analysis demonstrated the activation of immune- and inflammation-related pathways post-SCI, including NOD-like receptors, nuclear factor-kappa B(NF-κB), and Toll-like receptor (TLR) pathways. EA primarily influenced intestinal inflammation and autoimmune functions. 16S rDNA sequencing illustrated that EA did not alter the diversity of gut microbiota. However, EA altered the gut microbiota composition in SCI rats, increasing Lactobacillus and Akkermansia genera while rebalancing the Firmicutes/Bacteroidetes ratio. Furthermore, depletion of gut microbiota by antibiotics disrupted the intestinal barrier, reduced the expression of intestinal barrier proteins Zonula Occludens-1 (ZO-1) and Occludin, elevated serum lipopolysaccharide-binding protein (LBP) levels, exacerbated spinal cord tissue damage, and hindered motor function recovery in SCI rats. FMT from donors treated with EA reduced LBP levels in the intestine, blood, and spinal cord of rats, inhibited the TLR4 myeloid differentiation primary response protein 88 (MyD88)-NF‑κB pathway and NLRP3-dependent pyroptosis, and improved motor function. On the other hand, FMT treatment resulted in decreased body weight and food intake, whereas FMT using EA-treated donors effectively alleviated these alterations. ConclusionEA effectively alleviated neuroinflammatory responses in rats with SCI, primarily through regulating the gut microbiota and suppressing the NLRP3-dependent pyroptosis signaling pathway.

AIM To investigate the differences between boiled powder and decoction of Ermiao Powder.METHODS GC-MS was used to identify volatile constituents,after which the content determination of linalool,4-terpineol,α-terpineol,β-eucalyptol and taxifolin in distillate was performed,evaporation rate curve was drawn.The rat model for collagen-induced arthritis was established,then HE and SO/FG staining were conducted,and body weight,footpad swelling degree,arthritis score,immune organ(spleen,thymus)indices,serum inflammatory factors(IL-10,IL-6),ankle joint structure(foot claw swelling,micro-CT)and locomotor ability were detected.RESULTS Total 43 and 26 volatile constituents were identified in boiled powder and decoction,respectively.With the extension of boiling time,the average evaporation rates of 5 volatile constituents in the boiled powder distillate demonstrated the trends of first increase and then decrease,which reached a maximum at 15-20 min;those in the decoction distillate displayed the trends of decrease,which were not detected after 15 min except for β-eucalyptol.Compared with the decoction,the boiled powder exhibited stronger effects on improving foot swelling and arthritis score,alleviating pathological changes in joint tissues,inhibiting inflammatory factors and restoring motor ability.CONCLUSION More volatile constituents are observable in the boiled powder of Ermiao Powder than those in its decoction,along with stronger anti-rheumatoid arthritis activity.The optimal decocting endpoint is determined to be within 15 min in boiling water.

The telomere structure in the cell nucleus is crucial for maintaining the stability and functions of chromosomes.Telomerase is a ribonucleoprotein reverse transcriptase,which catalyzes the elongation of telomeres using its own RNA as a template,thereby counteracting the shortening of telomeres caused by chromosome replication and cell division.Due to its overexpression in over 85％of malignant tumor cells,telomerase has emerged as a highly promising biomarker and a novel target for cancer therapy.In recent years,given the importance of precise quantification of telomerase activity in guiding medical diagnosis and treatment strategies,researchers have developed various high-performance telomerase detection techniques.Among these,electrochemical biosensing technique has cause much attention due to its high sensitivity,operational convenience,rapid response,and ease of miniaturization.This paper focused on the latest advances in electrochemical sensing technique for detection of telomerase activity,aiming to provide inspiration for designing novel telomerase activity detection strategies by elucidating three unique properties of telomerase primer extension products.

Objective:To investigate the effect of laparoscopic high ligation of spermatic cord vein in patients with chronic prostatitis and varicocele prostatitis.Methods:A total of 90 varicocele patients were selected from January 2016 to December 2020,including 33 patients with chronic prostatitis.Changes of white blood cell count,National Institutes of Health chronic prostatitis symp-tom index(NIH-CPSI)score and serum testosterone level in the expressed prostate secretion(EPS)were observed before and after the operation of laparoscopic high ligation of spermatic vein.Results:All patients were followed up three months after the surgery.There was no significant difference in the white blood cell counts in EPS,NIH-CPSI score,and serum testosterone level in patients with vari-cocele-only who underwent high ligation surgery after the operation.However,the white blood cell count in the EPS of patients with chronic prostatitis was lower than that before 3 months of operation([12.39±4.23]× 10 9/L vs[21.36±5.05]× 10 9/L).The NIH-CPSI score was significantly lower than that before operation([12.71±6.21]vs[26.76±8.43]).And the serum testosterone level was higher than that before operation([4.34±1.77]ng/ml vs[2.36±1.05]ng/ml).Conclusion:Laparoscopic high liga-tion of the spermatic vein in patients with chronic prostatitis and varicocele could effectively reduce the number of white blood cells in the EPS,boost the level of serum testosterone and improves symptoms of chronic prostatitis.

support the diagnosis.The treatment of PHCC is mainly based on surgery.Due to the characteristics of intact capsule,surgical resection is relatively easy and the cure rate is higher than that of ordinary hepatocellular carcinoma,and the postoperative survival rate is relatively ideal.For unresectable PHCC,palliative treatment based on transcatheter arterial chemoembolization can be used.This article reviews the progress on diagnosis and treatment of PHCC in order to provide reference for clinical practice.

AIM To investigate the differences between boiled powder and decoction of Ermiao Powder.METHODS GC-MS was used to identify volatile constituents,after which the content determination of linalool,4-terpineol,α-terpineol,β-eucalyptol and taxifolin in distillate was performed,evaporation rate curve was drawn.The rat model for collagen-induced arthritis was established,then HE and SO/FG staining were conducted,and body weight,footpad swelling degree,arthritis score,immune organ(spleen,thymus)indices,serum inflammatory factors(IL-10,IL-6),ankle joint structure(foot claw swelling,micro-CT)and locomotor ability were detected.RESULTS Total 43 and 26 volatile constituents were identified in boiled powder and decoction,respectively.With the extension of boiling time,the average evaporation rates of 5 volatile constituents in the boiled powder distillate demonstrated the trends of first increase and then decrease,which reached a maximum at 15-20 min;those in the decoction distillate displayed the trends of decrease,which were not detected after 15 min except for β-eucalyptol.Compared with the decoction,the boiled powder exhibited stronger effects on improving foot swelling and arthritis score,alleviating pathological changes in joint tissues,inhibiting inflammatory factors and restoring motor ability.CONCLUSION More volatile constituents are observable in the boiled powder of Ermiao Powder than those in its decoction,along with stronger anti-rheumatoid arthritis activity.The optimal decocting endpoint is determined to be within 15 min in boiling water.

This study investigated the infection status,drug resistance,and molecular characteristics of Shiga toxin-producing Escherichia coli(STEC)in domestic goats in Chengkou county,Chongqing.In August 2023,283 fecal samples were collected from households in Chengkou county.After enrichment with EC broth and inoculation onto selective media,samples that tested positive for stx1/stx2 were selected for further isolation.The positive strains were investigated with antimicrobial susceptibility testing and whole genome sequencing.According to the whole genomic sequences,the stx subtypes,serotypes,multi-locus sequence types,virulence genes,drug resistance genes,and phylogenetic relationships of the STEC strains were analyzed.Forty-six strains of STEC were isolated from 283 goat fecal samples,thus resulting in a detection rate of 16.25%.The 46 STEC strains were categorized into 12 O∶H serotypes,among which O76∶H19 and O8∶H7 predominated,each represented by 9 strains.Five STEC strains were identified as serotype O157∶H7.The 46 STEC strains were categorized into 11 sequence types(STs),among which ST675 and ST196 predominated,each represented by nine strains,accounting for a 19.57%proportion.The strains were categorized into 7 stx subtypes,among which stx1c(26/46,56.52%),followed by stx2k(9/46,19.57%)predominated.All nine Stx2k-STEC strains were identified as serotype O8∶H7 and sequence type ST196.In antimicrobial susceptibility testing,2 STEC strains were resistant to ampicillin,one strain was resistant to ampicillin/sulbactam,one strain was resistant to cefazolin,and one strain was resistant to cefoxitin.Nine Stx2k-STEC strains were found to carry the beta-lactam resistance gene blaEC-18.Antimicrobial sensitivity tests revealed that the nine Stx2k-STEC strains were sensitive to all 15 tested antibiotics.Moreover,phylogenetic analysis indicated that the 9 Stx2k-STEC strains were remarkably similar but showed high genetic diversity with respect to that of the Stx2k-STEC strains isolated from other regions in China.Goatsare an important animal reservoir for STEC in theChengkou district of Chongqing,and novel sequence type Stx2k-STEC strains distinct from those found in other regions of China were identified in this region.

Aim To explore the mechanism of hispidu-lin in the treatment of cervical cancer by using network pharmacology and molecular docking methods and veri-fy it by in vitro experiments.Methods Cervical canc-er HeLa and SiHa cells were cultivated in vitro,and CCK-8 assay,cloning assay,scratch assay,transwell as-say,and flow cytometry were used to detect the effects of hispidulin on cell proliferation,migration,invasion,and apoptosis.SwissTarget Prediction was used to ob-tain predicted targets for hispidulin.Potential targets for cervical cancer were screened in GeneCards disease database.R software Venn package was used to obtain the intersection target genes of hispidulin and cervical cancer,STRING website and Cytoscape software were used to obtain protein-protein interaction(PPI)net-work,and the core targets were screened.The GEIPA data analysis platform was employed to analyze the dif-ferential gene expression levels of core targets in cervi-cal cancer.Gene Ontology(GO)and Kyoto Encyclo-pedia of Genes and Genomes(KEGG)enrichment a-nalysis were performed,and molecular docking was car-ried out on key targets.Western blot was used to detect the regulatory effects of hispidulin on the expression of key proteins PI3K,p-Akt,as well as core target pro-teins MMP9 and RARP1 in the PI3K/Akt signaling pathway.Results Cell experiments showed that after treatment with hispidulin,the proliferation and colony formation abilities of HeLa and SiHa cells significantly decreased in a concentration-and time-dependent man-ner.At the same time,the lateral and longitudinal mi-gration and invasion abilities of HeLa cells decreased,and the level of apoptosis significantly increased.A to-tal of 87 intersection targets between hispidulin and cervical cancer were obtained,and eight core targets,namely,Akt1,EGFR,SRC,ESR1,PTGS2,GSK3β,MMP9,and PARP1,were selected based on the degree values in network topology analysis.KEGG enrichment screening identified PI3K/Akt signaling pathway,canc-er pathway,and other signaling pathways.The molecu-lar docking results showed that hispidulin had strong affinity activity with AktⅠ,P13K,MMP9,and RARP1.Western blot results showed downregulation of PI3K,p-Akt expression,as well as MMP9 and RARP1 expres-sion.Conclusions Hispidulin can inhibit the prolif-eration,migration,invasion,and promote apoptosis of cervical cancer cells by downregulating the PI3K/Akt signaling pathway and the expression of MMP9 and RARP1.

AIM To investigate the repair effects of tauroursodeoxycholic acid(TUDCA)combined with bone marrow mesenchymal stem cells(BMSCs)transplantation on spinal cord injury(SCI)in rats.METHODS The rats were randomly divided into the sham operation group,the model group,the TUDCA group,the BMSCs transplantation group and the combination therapy of TUDCA and BMSCs transplantation group,with the SCI rat model established by Allen's method.The next day after modeling,the rats of TUDCA and combination therapy groups were given 200 mg/kg TUDCA by gavage.On the 3rd day after modeling,rats in BMSCs transplantation group and combination therapy group were injected with 1 mL tuned bone marrow BMSCs(the 3rd generation,1× 106/mL)via tail vein.Rats in the sham operation group and the model group were given gastric perfusion of normal saline and injection of 1 mL PBS through tail vein.On the 3rd,7th and 14th day after modeling,the rats had their motor function of hind limbs observed and BBB score determined.After the corresponding drug administration,the rats had their movement track of hind limbs recorded by footprint experiment;their the protein expressions of IL-6,IL-10,Arg-1,PI3K and Akt in spinal cord tissue detected by Western blot;their pathological changes of spinal cord tissue observed by HE staining and Nissl staining;and their expressions of MAP2,GAP43 and GFAP detected by immunofluorescence staining.RESULTS Compared with the model group,the groups intervened with TUDCA,or BMSCs transplantation,or combination therapy shared improved hind limb function and spinal cord histomorphology(P＜0.05);increased fluorescence intensity of MAP2 and GAP43,and protein expressions of IL-10,Arg-1,p-PI3K and p-Akt(P＜0.05);decreased fluorescence intensity of GFAP and IL-6 protein expressions(P＜0.05);among which the combination therapy group took the lead(P＜0.05).CONCLUSION The combination therapy of TUDCA and BMSCs transplantation may restore the function of the rat model of SCI by reducing inflammatory reaction,alleviating secondary injury,and promoting axon and myelin regeneration via PI3K/Akt signaling pathway.