Aim To reveal the mechanism of CIP4 homologs protein 1(RICH1)are involved in the regu-lation of myocardial fibrosis.Methods Mouse cardiac fibroblasts(MCFs)cells were treated with transforming growth factor-β(TGF-β1)to induce the formation of a myocardial fibrosis cell model;the level of the target protein was detected by Western blotting;and the RICH1 gene was detected by transfection of the cells with plasmid.The RICH1 gene was overexpressed(RICH 1 OE)using plasmid transfection;the RICH1 gene was silenced using siRNA fragment(siRICH1);and the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3 a1,and Acta2,were de-tected using RT-qPCR.Results RICH1 was signifi-cantly down-regulated in TGF-β1-treated MCFs;the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3a1,and Acta2,were down-regu-lated in the RICH1 OE+TGF-β1 group;and in the siRICH1+TGF-β1 group,myocardial fibrosis marker genes,such as Col1 a1,Col3a1 and Acta2 were up-regulated at the expression level;phosphorylated SMAD2(p-SMAD2)and phosphorylated SMAD3(p-SMAD3)levels were down-regulated in the siRICH1 OE+TGF-β1 group.p-SMAD2 and P-SMAD3 levels were upregulated in the siRICH1+TGF-β1 group.Conclusion RICH1 inhibits TGF-β1-induced myo-cardial fibrosis;RICH1 inhibits TGF-β1-induced myo-cardial fibrosis by negatively regulating the SMAD2/3 signaling pathway.

Aim To investigate the effect of high-fat diet on the tight junction function injury of Sertoli cells through the NF-κB/NLRP3 signaling pathway in mice and to explore the underlying mechanism.Methods Male C57BL/6J mice were fed with high-fat or normal diet for five months.The body and gonadal organ weight of mice were measured,and their indices were calculated.The sperm concentration,the sperm viabili-ty,the testicular histomorphology and the expression levels of tight junction proteins ZO-1,Occludin and Claudin-11 were measured.TM4 cells were treated with palmitic acid(PA)for 24 h.Cell viability was detected by CCK-8 method.Then,TM4 cells were di-vided into different groups treated with PA(0,50,100,200 and 300 μmnol·L-1),and the expression lev-els of tight junction proteins ZO-1,Occludin and Clau-din-11 were detected by Western blot.The tight junc-tion permeability of TM4 cells were detected by transepithelial electrical resistance(TEER)and FITC-dextran.The expression levels of mRNA and proteins for the NF-κB/NLRP3 pathway-related factors were de-tected by RT-qPCR and Western blot.Results The results from animal experiments showed that high-fat diet increased body weight and seminal vesicle weight of mice,and decreased testicular index,epididymal in-dex,sperm concentration and sperm motility of mice.High-fat diet also caused testicular tissue structure damage and down-regulated the expression levels of tight junction proteins ZO-1 and Occludin,without af-fecting the expression of Claudin-11.In vitro,PA sig-nificantly down-regulated the expression levels of ZO-1,Occludin and Claudin-11 in TM4 cells,increased the cell permeability,as well as up-regulated the mRNA and protein expression levels of NLRP3/NF-κB signa-ling pathway-related factors in TM4 cells.Conclusions High-fat diet can impair the function of tight junction of testicualr Sertoli cells,and the machanism may be related to the activation of the NF-κB/NLRP3 signaling pathway,resulting in Sertoli cell inflammation in mice.

Objective:To isolate Yersinia enterocolitica phage and study its biological and genomic characteristics, and explore its application value. Methods:Phages were isolated from cecum of Apodemus chevrieri in the plague focus of wild rodent in Lijiang employing Yersinia enterocolitica (CMCC52301) as the host strain. After purification, proliferation and concentration, the morphology was observed by transmission electron microscope, and the host spectrum of the phage was determined by double-layer plate method. Phage DNA was extracted and sequenced, and the raw sequences data were assembled and visually analyzed, also performed genome functional annotation, phage classification status, phylogenetic tree, genome collinearity analysis, etc. Results:One strain of Yersinia enterocolitica phage was isolated and named vB_Yen_YN301-151, which had a head and a contractile tail. The phage only lysed Yersinia enterocolitica, and could not lyse other tested strains. And at a lysis temperature of 37 ℃, more transparent plaques were observed compared to at a lysis temperature of 21 ℃. The gene structure of this phage was double-stranded DNA, with a genome length of 51 176 bp and a GC content of 46.96%, and 95 genes were predicted to encode 15 structural morphological related proteins, 9 proteins involved in replication and metabolism, 3 DNA assembly functional proteins, 1 phage lysis related protein, and 67 hypothetical proteins. It was identified that the phage belong to the Drexlerviridae family of the Caudoviricetes class, clustered with vB_YenM_534, vB_Yen_X1, vB_YenM_281, and vB_YenM_531 in the phylogenetic tree and exhibited good collinearity with PY100, vB_YenM_ 25, and vB_Yen_X1. Conclusions:The successfully isolated vB_Yen_YN301-151 is a novel phage belonging to the Drexlerviridae family, and its gene encoded proteins are mostly hypothetical proteins. It only lyses Yersinia enterocolitica, with host specificity and has the potential to be used as diagnostic phage for screening Yersinia enterocolitica.

Aim To reveal the mechanism of CIP4 homologs protein 1(RICH1)are involved in the regu-lation of myocardial fibrosis.Methods Mouse cardiac fibroblasts(MCFs)cells were treated with transforming growth factor-β(TGF-β1)to induce the formation of a myocardial fibrosis cell model;the level of the target protein was detected by Western blotting;and the RICH1 gene was detected by transfection of the cells with plasmid.The RICH1 gene was overexpressed(RICH 1 OE)using plasmid transfection;the RICH1 gene was silenced using siRNA fragment(siRICH1);and the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3 a1,and Acta2,were de-tected using RT-qPCR.Results RICH1 was signifi-cantly down-regulated in TGF-β1-treated MCFs;the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3a1,and Acta2,were down-regu-lated in the RICH1 OE+TGF-β1 group;and in the siRICH1+TGF-β1 group,myocardial fibrosis marker genes,such as Col1 a1,Col3a1 and Acta2 were up-regulated at the expression level;phosphorylated SMAD2(p-SMAD2)and phosphorylated SMAD3(p-SMAD3)levels were down-regulated in the siRICH1 OE+TGF-β1 group.p-SMAD2 and P-SMAD3 levels were upregulated in the siRICH1+TGF-β1 group.Conclusion RICH1 inhibits TGF-β1-induced myo-cardial fibrosis;RICH1 inhibits TGF-β1-induced myo-cardial fibrosis by negatively regulating the SMAD2/3 signaling pathway.

Aim To investigate the effect of high-fat diet on the tight junction function injury of Sertoli cells through the NF-κB/NLRP3 signaling pathway in mice and to explore the underlying mechanism.Methods Male C57BL/6J mice were fed with high-fat or normal diet for five months.The body and gonadal organ weight of mice were measured,and their indices were calculated.The sperm concentration,the sperm viabili-ty,the testicular histomorphology and the expression levels of tight junction proteins ZO-1,Occludin and Claudin-11 were measured.TM4 cells were treated with palmitic acid(PA)for 24 h.Cell viability was detected by CCK-8 method.Then,TM4 cells were di-vided into different groups treated with PA(0,50,100,200 and 300 μmnol·L-1),and the expression lev-els of tight junction proteins ZO-1,Occludin and Clau-din-11 were detected by Western blot.The tight junc-tion permeability of TM4 cells were detected by transepithelial electrical resistance(TEER)and FITC-dextran.The expression levels of mRNA and proteins for the NF-κB/NLRP3 pathway-related factors were de-tected by RT-qPCR and Western blot.Results The results from animal experiments showed that high-fat diet increased body weight and seminal vesicle weight of mice,and decreased testicular index,epididymal in-dex,sperm concentration and sperm motility of mice.High-fat diet also caused testicular tissue structure damage and down-regulated the expression levels of tight junction proteins ZO-1 and Occludin,without af-fecting the expression of Claudin-11.In vitro,PA sig-nificantly down-regulated the expression levels of ZO-1,Occludin and Claudin-11 in TM4 cells,increased the cell permeability,as well as up-regulated the mRNA and protein expression levels of NLRP3/NF-κB signa-ling pathway-related factors in TM4 cells.Conclusions High-fat diet can impair the function of tight junction of testicualr Sertoli cells,and the machanism may be related to the activation of the NF-κB/NLRP3 signaling pathway,resulting in Sertoli cell inflammation in mice.

Objective:To isolate Yersinia enterocolitica phage and study its biological and genomic characteristics, and explore its application value. Methods:Phages were isolated from cecum of Apodemus chevrieri in the plague focus of wild rodent in Lijiang employing Yersinia enterocolitica (CMCC52301) as the host strain. After purification, proliferation and concentration, the morphology was observed by transmission electron microscope, and the host spectrum of the phage was determined by double-layer plate method. Phage DNA was extracted and sequenced, and the raw sequences data were assembled and visually analyzed, also performed genome functional annotation, phage classification status, phylogenetic tree, genome collinearity analysis, etc. Results:One strain of Yersinia enterocolitica phage was isolated and named vB_Yen_YN301-151, which had a head and a contractile tail. The phage only lysed Yersinia enterocolitica, and could not lyse other tested strains. And at a lysis temperature of 37 ℃, more transparent plaques were observed compared to at a lysis temperature of 21 ℃. The gene structure of this phage was double-stranded DNA, with a genome length of 51 176 bp and a GC content of 46.96%, and 95 genes were predicted to encode 15 structural morphological related proteins, 9 proteins involved in replication and metabolism, 3 DNA assembly functional proteins, 1 phage lysis related protein, and 67 hypothetical proteins. It was identified that the phage belong to the Drexlerviridae family of the Caudoviricetes class, clustered with vB_YenM_534, vB_Yen_X1, vB_YenM_281, and vB_YenM_531 in the phylogenetic tree and exhibited good collinearity with PY100, vB_YenM_ 25, and vB_Yen_X1. Conclusions:The successfully isolated vB_Yen_YN301-151 is a novel phage belonging to the Drexlerviridae family, and its gene encoded proteins are mostly hypothetical proteins. It only lyses Yersinia enterocolitica, with host specificity and has the potential to be used as diagnostic phage for screening Yersinia enterocolitica.

BACKGROUND: Asthma is a common chronic inflammatory airway disease and intermittent hypoxia is increasingly recognized as a factor that may impact disease progression. The present study investigated whether intermittent hypoxia (IH) could aggravate asthma by promoting hypoxia-inducible factor-1α (HIF-1α)/nucleotide-binding oligomerization domain (NOD)-like receptor pyrin domain-containing protein 3 (NLRP3)/interleukin (IL)-1β-dependent pyroptosis and the inflammatory response and further elucidated the underlying molecular mechanisms involved. METHODS: A total of 49 patients diagnosed with severe bronchial asthma and diagnosed by polysomnography were enrolled at Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, between January 2022 and December 2022, and their general data and induced sputum were collected. BEAS-2B cells were treated with IL-13 and subjected to IH. An ovalbumin (OVA)-treated mouse model was also used to assess the effects of chronic intermittent hypoxia (CIH) on asthma. Pyroptosis, the inflammatory response, and related signalling pathways were assessed in vivo and in vitro . RESULTS: In this study, as the apnoea and hypopnea index (AHI) increased, the proportion of patients with uncontrolled asthma increased. The proportions of neutrophils and the levels of IL-6, IL-8, HIF-1α and NLRP3 in induced sputum were related to the AHI. NLRP3-mediated pyroptosis, which could be mediated by the HIF-1α signalling pathway, was activated in IL-13 plus IH-treated BEAS-2B cells and in the lungs of OVA/CIH mice. HIF-1α downregulation significantly reduced lung pyroptosis and ameliorated neutrophil inflammation by modulating the NLRP3/IL-1β pathway both in vitro and in vivo . Similarly, pretreatment with LW6, an inhibitor of HIF-1α, effectively blocked the generation of inflammatory cytokines in neutrophils. In addition, administration of the NLRP3 activator nigericin obviously increased lung neutrophil inflammation. CONCLUSIONS Obstructive sleep apnoea-hypopnea syndrome (OSAHS) is a risk factor for asthma exacerbation. IH aggravates neutrophil inflammation in asthma via NLRP3/IL-1β-dependent pyroptosis mediated by the HIF-1α signalling pathway, which should be considered a potential therapeutic target for the treatment of asthma with OSAHS.
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Humans ; Asthma/metabolism* ; Animals ; Pyroptosis/physiology* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Mice ; Signal Transduction/physiology* ; Male ; Hypoxia/metabolism* ; Female ; Interleukin-1beta/metabolism* ; Adult ; Inflammation/metabolism* ; Middle Aged ; Mice, Inbred C57BL

Background: Condyloma acuminatum (CA) is a common sexually transmitted disease caused by human papillomavirus (HPV). In recent years, research on anal CA has primarily focused on treatment rather than underlying mechanisms. The mechanism of HPV persistence and recurrence in CA require further exploration. It needs multiple researches in mechanisms to focalize treatment targets. Objective: To investigate the relationship between intestinal mucosal immunity and the relapse of anal CA and persistent infection. Methods: Levels of interferon gamma (IFN-γ) and secretory immunoglobulin A (sIgA) were measured using enzyme-linked immunosorbnent assay in anal mucosal cells obtained from patients treated at Tianjin Union Medical Center from September 2022 to December 2024. All the participants signed Informed Consent and the whole plan was approved by Institutional Review Board in Tianjin Union Medical Center (No. B155). Results: The levels of IFN-γ and sIgA significantly decreased after infection, and persistent infection exhibited even lower levels. These two factors increased following treatment, reaching peak concentrations at 4 weeks before decreasing again. Conclusion These findings demonstrate a significant association between persistent anal CA infection and dysregulation of intestinal mucosal immunity.

Objective:To systematically evaluate the clinical efficacy and safety of Tonifying kidney and activating blood thera-py for the treatment of diabetic mellitus erectile dysfunction.Methods:China National Knowledge Infrastructure(CNKI),Wanfang Data,VIP,Chinese Biomedical Database(CBM),PubMed,Cochrane Library,Embase and Web of Science were searched from incep-tion until October 20th of 2024,for randomized controlled trials of Tonifying kidney and activating blood therapy for the treatment of dia-betic erectile dysfunction.Literature screening,quality evaluation,and data extraction were carried out in accordance with relevant standards.The software of RevMan5.4 was used for the analysis of publication bias.And meta-analysis was conducted to assess the im-pact of this therapy on IIEF-5,total effective rate,adverse reactions.The evidence levels according to the analysis results were evalua-ted.Results:Totally 19 RCTs were included,involving 1 612 patients.The result of meta-analysis indicated that Tonifying kidney and activating blood therapy had advantages on the improvement of IIEF-5 scores(MD=3.59,95％CI[2.14,5.03],P＜0.01),total effective rate(OR=4.30,95％CI[3.29,5.32],P＜0.000 01).However,there was no statistically significant difference in the inci-dence of adverse reactions(OR=0.98,95％CI[0.48,2.01],P=0.96)between the two groups.Conclusions:Tonifying kidney and activating blood therapy can improve the clinical efficacy and IIEF-5 score for the patients with diabetic erectile dysfunction.But considering the limited quantity of included studies,more high-quality studies still be needed to validate the therapeutic effect.

Objective:To explore the mechanism of hypertension inducing erectile dysfunction(ED)using transcriptomics and network pharmacology.Methods:We randomly divided 12 male rats with spontaneous hypertension(SHT)into an L-arginine(LA)group(n=6)and an SHT model control(MC)group(n=6),took another 6 Wistar Kyoto male rats as normal controls(NC),and treated the animals in the LA group by intraperitoneal injection of LA at 400 mg/kg and those in the latter two groups with physio-logical saline,once a day,all for 7 days.Then we observed the blood pressure and penile erection of the rats,and determined the ex-pressions of the cGMP/PKG signaling pathway-related proteins and mRNAs in different groups using ELISA,Western blot and RT-qPCR.Results:Transcriptomics combined with network pharmacology showed that the cGMP/PKG signaling pathway played a key role in hypertension-induced ED.In vivo animal experiments revealed a significantly lower frequency of penile erections in the MC than in the NC group(1.33±0.52 vs 2.67±0.51,P＜0.05).The protein expressions of eNOS,PKG and sGC were markedly de-creased in the model controls compared with those the normal controls(P＜0.05),but remarkably upregulated in the LA group com-pared with those in the MC group(P＜0.05).Conclusion:Hypertension decreases the expressions of eNOS,NO,sGC,cGMP and PKG proteins and the level of testosterone by inhibiting the cGMP/PKG signaling pathway,which consequently suppresses the relaxa-tion of the penile vascular smooth muscle and reduces erectile function.