ObjectiveTo explore the potential mechanism of Xiezhuo Jiedu prescription in regulating autophagy in the treatment of ulcerative colitis （UC） by bioinformatics and animal experiments. MethodsThe differentially expressed genes （DEGs） in the colonic mucosal tissue of UC patients was obtained from the Gene Expression Omnibus （GEO）， and those overlapped with autophagy genes were obtained as the differentially expressed autophagy-related genes （DEARGs）. DEARGs were imported into Metascape and STRING， respectively， for gene ontology/Kyoto Encyclopedia of Genes and Genomics （GO/KEGG） enrichment analysis and protein-protein interaction （PPI） analysis. Finally， 15 key DEARGs were obtained. The core DEARGs were obtained by least absolute shrinkage and selection operator （LASSO） regression and receiver operating characteristic curve （ROC） analysis. The CIBERSORT deconvolution algorithm was used to analyze the immunoinfiltration of UC patients and the correlations between core DEARGs and immune cells. C57BL/6J mice were assigned into a normal group and a modeling group. The mouse model of UC was established by free drinking of 2.5% dextran sulfate sodium. The modeled mice were assigned into low-， medium-， and high-dose Xiezhuo Jiedu prescription and mesalazine groups according to the random number table method and administrated with corresponding agents by gavage for 7 days. The colonic mucosal morphology was observed by hematoxylin-eosin staining. The protein and mRNA levels of cysteinyl aspartate-specific proteinase 1 （Caspase-1）， cathepsin B （CTSB）， C-C motif chemokine-2 （CCL2）， CXC motif receptor 4 （CXCR4）， and hypoxia-inducing factor-1α （HIF-1α） in the colon tissue were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction， respectively. ResultsThe dataset GSE87466 was screened from GEO and interlaced with autophagy genes. After PPI analysis， LASSO regression， and ROC analysis， the core DEARGs （Caspase-1， CCL2， CTSB， and CXCR4） were obtained. The results of immunoinfiltration analysis showed that the counts of NK cells， M0 macrophages， M1 macrophages， and dendritic cells in the colonic mucosal tissue of UC patients had significant differences， and core DEARGs had significant correlations with these immune cells. This result， combined with the prediction results of network pharmacology， suggested that the HIF-1α signaling pathway may play a key role in the regulation of UC by Xiezhuo Jiedu prescription. The animal experiments showed that Xiezhuo Jiedu prescription significantly alleviated colonic mucosal inflammation in UC mice. Compared with the normal group， the model group showed up-regulated protein and mRNA levels of caspase-1， CCL2， CTSB， CXCR4， and HIF-1α， which were down-regulated after treatment with Xiezhuo Jiedu prescription or mesalazine. ConclusionCaspase-1， CCL2， CTSB， and CXCR4 are autophagy genes that are closely related to the onset of UC. Xiezhuo Jiedu prescription can down-regulate the expression of core autophagy genes to alleviate the inflammation in the colonic mucosa of mice.

Objective To investigate the mechanism of Kuijie Ankang Decoction in regulating immune,autophagy and intestinal flora in the treatment of ulcerative colitis(UC).Methods UC mouse model was established by free drinking with sodium dextran sulfate.The mice were randomly divided into blank control group,model group,Kuijie Ankang Decoction group,salazine sulfopyridine(SASP)group and 3-methyladenine(3-MA)group,with 12 mice in each group.Each drug group was given corresponding drugs for gavage,the blank control group and model group were given the same volume of distilled water for gavage for 7 days.The general condition of mice was observed and the disease activity index(DAI)was scored,the morphology of colon tissue was observed by HE staining,the contents of TNF-α,IL-1β,IL-6,IL-8 and IL-10 in colon tissue were detected by ELISA,the mRNA and protein expressions of LC3,Beclin-1 and p62 in colon tissue were detected by RT-qPCR and Western blot,respectively.16S rDNA sequencing was used to analyze the structure of intestinal flora.Results Compared with the blank control group,the mice in the model group showed a decrease in body mass,an increase in DAI score,a decrease in colon length,serious mucosal injury and inflammatory cell infiltration,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue significantly increased(P<0.05,P<0.01),the content of IL-10 decreased(P<0.01),the mRNA expressions of LC3 and Beclin-1 in colon tissue decreased(P<0.05,P<0.01),the mRNA and protein expression of p62 increased(P<0.01),while the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins decreased(P<0.05,P<0.01).Compared with the model group,the body mass of mice in Kuijie Ankang Decoction group and SASP group increased(P<0.05),DAI score decreased(P<0.05),the colon length increased(P<0.05),the pathological damage of colon mucosa was alleviated,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue decreased(P<0.05,P<0.01),the content of IL-10 increased(P<0.01),the expressions of LC3 and Beclin-1 mRNA in colon tissue increased(P<0.05,P<0.01),the expression of p62 mRNA and protein decreased(P<0.05,P<0.01),the expressions of LC3Ⅱ/LC3Ⅰand Beclin-1 protein increased(P<0.05,P<0.01).16S rDNA sequencing results showed that the diversity and evenness of the intestinal flora in the model group mice decreased,with a decrease in the relative abundance of Firmicutes,Actinobacteria and Patescibacteria(P<0.05),and an increase in the relative abundance of Bacteroidota,Verrucomicrobiota and Proteobacteria(P<0.05);the relative abundance of Bacilli and Coriobacteriia decreased(P<0.05),the relative abundance of Bacteroidia,Clostridia and Verrucomicrobiae increased(P<0.05);the relative abundance of Ligilactobacillus and Dubosiella decreased(P<0.05),the relative abundance of unclassified Muribaculaceae,Lachnospiraceae NK4A136_group,Akkermansia and unclassified Lachnospiraceae increased(P<0.05).Compared with the model group,the diversity and evenness of intestinal flora increased in Kuijie Ankang Decoction group and SASP group,with an increase in the relative abundance of Firmicutes(P<0.05),a decrease in the relative abundance of Bacteroidota and Verrucomicrobiota(P<0.05),the relative abundance of Bacteroidia and Bacilli increased(P<0.05),the relative abundance of Verrucomicrobiae decreased(P<0.05);the relative abundance of unclassified Muribaculaceae and Ligilactobacillus increased(P<0.05),while the relative abundance of Lachnospiraceae NK4A136_group and Akkermansia decreased(P<0.05).Conclusion Kuijie Ankang Decoction can significantly improve the intestinal mucosal injury of UC mice,and the mechanism may be related to the regulation of colon autophagy level and intestinal flora disorder.

Colorectal inflammatory cancer transformation poses a major risk to patients with colitis.Patients with chronic intestinal inflammation have an approximately 2-3 fold increased risk of developing colorectal cancer(CRC).Unfortunately,there is currently no effective intervention available.Huangqin decoction(HQD),a well-known traditional Chinese medicine(TCM)formula,is frequently clinically prescribed for treating patients with colitis,and its active ingredients have effective antitumour efficacy.Nonetheless,the mechanism of HQD-mediated prevention of colorectal inflammatory cancer transformation remains unclear.A strategy integrating metagenomic,lipidomic,and messenger RNA(mRNA)sequencing analysis was used to investigate the regulatory effects of HQD on the gut microbiome,metabolism and potential mechanisms involved in colorectal inflammatory cancer transformation.Our study revealed that HQD suppressed colorectal inflammatory cancer transformation,which was associated with enhanced in-testinal barrier function,decreased the inflammatory response,and regulation of the gut microbiome.Notably,cohousing experiments revealed that the transfer of the gut microbiome from HQD-treated mice largely inhibited the pathological transformation of colitis.Moreover,gut microbiome transfer from HQD-treated mice primarily resulted in the altered regulation of fatty acid metabolism,especially the remodeling of arachidonic acid metabolism,which was associated with the amelioration of pathological transformation.Arachidonic acid metabolism and the key metabolic enzyme arachidonic acid 12-lipoxygenase(ALOX12)were affected by HQD treatment,and no obvious protective effect of HQD was observed in Alox12-/-mice,which revealed that ALOX12 was a critical mediator of HQD protection against colorectal inflammatory cancer transformation.In summary,multiple omics analyses were applied to produce valuable data and theoretical support for the application of HQD as a promising intervention for the transformation of inflammatory CRC.

Objective To investigate the mechanism of Kuijie Ankang Decoction in regulating immune,autophagy and intestinal flora in the treatment of ulcerative colitis(UC).Methods UC mouse model was established by free drinking with sodium dextran sulfate.The mice were randomly divided into blank control group,model group,Kuijie Ankang Decoction group,salazine sulfopyridine(SASP)group and 3-methyladenine(3-MA)group,with 12 mice in each group.Each drug group was given corresponding drugs for gavage,the blank control group and model group were given the same volume of distilled water for gavage for 7 days.The general condition of mice was observed and the disease activity index(DAI)was scored,the morphology of colon tissue was observed by HE staining,the contents of TNF-α,IL-1β,IL-6,IL-8 and IL-10 in colon tissue were detected by ELISA,the mRNA and protein expressions of LC3,Beclin-1 and p62 in colon tissue were detected by RT-qPCR and Western blot,respectively.16S rDNA sequencing was used to analyze the structure of intestinal flora.Results Compared with the blank control group,the mice in the model group showed a decrease in body mass,an increase in DAI score,a decrease in colon length,serious mucosal injury and inflammatory cell infiltration,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue significantly increased(P<0.05,P<0.01),the content of IL-10 decreased(P<0.01),the mRNA expressions of LC3 and Beclin-1 in colon tissue decreased(P<0.05,P<0.01),the mRNA and protein expression of p62 increased(P<0.01),while the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins decreased(P<0.05,P<0.01).Compared with the model group,the body mass of mice in Kuijie Ankang Decoction group and SASP group increased(P<0.05),DAI score decreased(P<0.05),the colon length increased(P<0.05),the pathological damage of colon mucosa was alleviated,the contents of TNF-α,IL-1β,IL-6 and IL-8 in colon tissue decreased(P<0.05,P<0.01),the content of IL-10 increased(P<0.01),the expressions of LC3 and Beclin-1 mRNA in colon tissue increased(P<0.05,P<0.01),the expression of p62 mRNA and protein decreased(P<0.05,P<0.01),the expressions of LC3Ⅱ/LC3Ⅰand Beclin-1 protein increased(P<0.05,P<0.01).16S rDNA sequencing results showed that the diversity and evenness of the intestinal flora in the model group mice decreased,with a decrease in the relative abundance of Firmicutes,Actinobacteria and Patescibacteria(P<0.05),and an increase in the relative abundance of Bacteroidota,Verrucomicrobiota and Proteobacteria(P<0.05);the relative abundance of Bacilli and Coriobacteriia decreased(P<0.05),the relative abundance of Bacteroidia,Clostridia and Verrucomicrobiae increased(P<0.05);the relative abundance of Ligilactobacillus and Dubosiella decreased(P<0.05),the relative abundance of unclassified Muribaculaceae,Lachnospiraceae NK4A136_group,Akkermansia and unclassified Lachnospiraceae increased(P<0.05).Compared with the model group,the diversity and evenness of intestinal flora increased in Kuijie Ankang Decoction group and SASP group,with an increase in the relative abundance of Firmicutes(P<0.05),a decrease in the relative abundance of Bacteroidota and Verrucomicrobiota(P<0.05),the relative abundance of Bacteroidia and Bacilli increased(P<0.05),the relative abundance of Verrucomicrobiae decreased(P<0.05);the relative abundance of unclassified Muribaculaceae and Ligilactobacillus increased(P<0.05),while the relative abundance of Lachnospiraceae NK4A136_group and Akkermansia decreased(P<0.05).Conclusion Kuijie Ankang Decoction can significantly improve the intestinal mucosal injury of UC mice,and the mechanism may be related to the regulation of colon autophagy level and intestinal flora disorder.

Objective:To observe the effect of reconstruction of Achilles insertion with Suture bridge technology after extirpation of avulsed bones block in the treatment of Beavis type Ⅲ avulsion fracture of the calcaneal tuberosity.Methods:The retrospective analysis was used. From January 2013 to January 2023, 78 patients with Beavis type Ⅲ avulsion fracture of the calcaneal tuberosity, treated in the Department 1 of Foot and Ankle and Department 2 of Foot and Ankle of the second Hospital of Tangshan were selected as research objects. According to different operation performed, 41 patients with the reconstruction of Achilles insertion with Suture bridge technology after extirpation of avulsed bones block were divided into the observation group and 37 patients with the open reduction internal fixation (ORIF) were divided into the control group. The delayed wound healing rate and the Haglund malformation rate, Maryland foot score, the American Orthopedic Foot and ankle Society (AOFAS) ankle-hindfoot score, the pain Visual Analogue Scale/Score (VAS) score, the Victorian Institute of Sports Assessment-Achilles (VISA-A) score, Arner-Lindholm scale of One-year postoperative were compared between these two groups. The measurement data with normal or approximate normal distribution were analyzed using t test, count data using chi-square test for comparison of between groups.Results:One year after surgery, the incidence of Haglund malformation in the observation group was 4.88% (2/41), which was lower than the control group's 29.73% (11/37). The difference between the two groups was statistically significant ( χ2=8.65,P=0.003). The excellent and good rate of Maryland foot function assessment in the observation group was 85.37% (35/41) higher than that in the control group (56.76% (21/37), and the difference between the two groups was statistically significant ( χ2=7.86, P=0.005). The AOFAS ankle hind foot score of the observation group ((90.44±6.66) points) was higher than that of the control group ((82.84±7.43) points), and the difference between the two groups was statistically significant ( t=4.77, P<0.001). The pain score of the observation group ((1.51±1.05) points) was lower than that of the control group ((2.95±1.13) points), and the difference between the two groups was statistically significant ( t=-5.81, P<0.001). The Achilles tendon score of the observation group ((81.05±5.87) points) was higher than that of the control group ((71.62±8.60) points), and the difference between the two groups was statistically significant ( t=5.70, P<0.001). The excellent and good rate of Arner Lindholm treatment efficacy evaluation for Achilles tendon in the observation group was 87.80% (36/41), which was higher than that in the control group (67.57% (25/37)), and the difference between the groups was statistically significant. Conclusion:The treatment of Beavis Ⅲ type calcaneal nodule avulsion fracture by removing the bone fragment can simultaneously remove the hypertrophic osteophyte, hardened bone, and Haglund deformity of the calcaneus, and clean the degenerated Achilles tendon and inflammatory tissue around the insertion point; The use of suture bridge technology to reconstruct the Achilles tendon insertion point has the advantages of high fixed strength, allowing early functional exercise, avoiding secondary removal of internal fixation, and achieving satisfactory therapeutic effects, which is worthy of clinical promotion.

Objective This study aims to develop an early in-hospital temperature management protocol for heat stroke patients and assess its effectiveness,providing guidance for rapid cooling and precise target temperature control.Methods The protocol was developed through a Delphi expert consultation combined with expert panel meetings.A multi-center,non-randomized,historical control study was conducted,utilizing convenience sampling to select heat stroke patients from the emergency departments of 7 tertiary hospitals in Zhejiang Province,China,between June and August 2024 as an experimental group.The protocol was implemented in this group,while the control group consisted of heat stroke patients treated between June and August 2022,prior to protocol implementation.Cooling rates,target temperature attainment rates,and clinical outcomes were compared between the 2 groups.Results The final protocol included 6 primary indicators,23 secondary indicators,and 56 tertiary indicators.After protocol implementation,the experimental group achieved a cooling rate of 0.08(0.05～0.09)℃/min within 0.5 hours,significantly higher than the control group,which had a rate of 0.04(0.02～0.06)℃/min(P＜0.001).The target temperature attainment rates at 0.5 hours and 2.0 hours were 55.93％and 98.31％,respectively,significantly higher than the rates of 15.87％and 61.11％in the control group(P＜0.001).The mechanical ventilation rate,hospitalization rate,ICU admission rate,and mortality rate in the experimental group were 25.42％,61.02％,44.07％,and 8.47％,respectively.Logistic regression analysis revealed that the early in-hospital temperature management protocol significantly reduced the risk of mechanical ventilation and hospitalization in heat stroke patients,with odds ratios(ORs)of 0.294 and 0.300,respectively(both P＜0.05).Conclusion The developed protocol for early in-hospital temperature management in heat stroke patients is scientific,systematic,and practical.It improves cooling rates and target temperature attainment,thereby enhancing the prognosis of heat stroke patients.

This study aims to prepare monoclonal antibody to S1 protein of porcine epidemic diar-rhea virus(PEDV).E.coli expression system and affinity chromatography were used to success-fully obtain purified recombinant PEDV S1 protein.After immunizing BALB/c mice,hybridoma technology and indirect ELISA were used to prepare and screen positive hybridoma cells.Finally,ascites antibodies were prepared by in vivo induction method.ELISA results showed that a total of 4 hybridoma cell lines with anti-PEDV S1 monoclonal antibody were screened,and they were named E6,G3,H6 and F2.The supernatant titers of all 4 hybridoma cell lines reached 1∶6 400.The monoclonal antibody H6 with higher antibody titers and more stable antibody secretion was selected for antibody type identification.It was found that monoclonal antibody H6 belongs to the IgG1 subclass and the light chain is the λ chain.The antibody titers that induced mouse ascites were 1∶106 and without cross-reaction with other proteins.Western blot results showed that the monoclonal antibody exhibited specific bands at 38 kDa with the recombinant S1 protein,PEDV QY2016,and PEDV CV777 strains.The IFA results also showed that the monoclonal antibody reacted with cells infected with PEDV QY2016 and PEDV CV777 strains,exhibiting a green fluo-rescent signal.The affinity constant of monoclonal antibody H6 was K=1.75×107 moL/L,indica-ting that the H6 strain had a good affinity and could be used for the development of subsequent di-agnostic antibodies.In summary,this study successfully prepared monoclonal antibodies that can specifically recognize PEDV S1 protein,which can be used for the antigen detection of PEDV and providing important test materials for the research of PEDV detection methods.

IgA vasculitis is a common childhood vasculitis disorder.Its primary clinical manifestation is cutaneous purpura,which is often accompanied by gastrointestinal and joint symptoms.Renal involvement can present as hematuria or proteinuria.This condition is recurrent and protracted,significantly affecting the health of children.The pathogenic factors in pediatric IgA vasculitis are diverse."Wind-toxin damaging collaterals"is presented as the core pathogenesis based on clinical practice and theoretical exploration.This concept essentially entails the latent attack of wind-toxin,collateral damage leading to blood extravasation,and healthy qi deficiency and lingering toxins,with the disease primarily located in the collateral vessels.The disease course is divided into three stages:acute,lingering,and recovery.Clinical practice should adhere to the pathogenesis principles and apply stage-based pattern differentiation and treatment.The acute stage involves wind-toxin attacking collaterals with dampness-heat accumulation.Treatment focuses on dispersing wind,clearing heat,and eliminating dampness to expel wind-toxin from the muscular exterior.The self-prescribed Qufeng Xiaodian Formula is a frequently selected modified formula.The lingering stage features latent wind-toxin in the collaterals and congealed accumulations in the kidney collaterals.Treatment aims to resolve wind-toxin,disperse stasis,and eliminate accumulation to remove densely accumulated wind-toxin.A modified Taohong Siwu Decoction is selected.The recovery stage involves deficient healthy qi with persistent wind assault and lingering toxins.Emphasis should be placed on cultivating the fundamental,strengthening the root,and nourishing yin,as well as supporting the healthy qi to dispel wind and expel toxins,while conditioning the body to prevent recurrence.A modified Guomin Decoction is selected.Clinical application emphasizes the combination of characteristic medicines,such as wind medicines to expel pathogens,insect medicines to identify collaterals,and vine medicines to unblock the meridians,to enhance the effects of identifying wind,resolving toxins,unblocking collaterals,and dispersing congealed accumulations.This study aims to systematically elaborate on the stage-based treatment strategy from the perspective of"wind-toxin damaging collaterals,"providing a theoretical basis and clinical practice reference for the traditional Chinese medicine diagnosis and treatment of pediatric IgA vasculitis.

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 explore the therapeutic effect of Shenwuyishen tablets(SWYST)in treating chronic kidney disease(CKD)and the underlying mechanism.Methods The pharmacological action of SWYST was evaluated in folic acid(FA)-induced mouse models by levels of serum creatinine and blood urea nitrogen,in-dexes of tubulointerstitial inflammation and tubular in-jury,and degrees of renal fibrosis.Differential genes from 22 types of cells in kidney tissue of CKD were i-dentified by single-cell and single-nuclei RNA sequen-cing datasets.Network pharmacology was used to pre-dict key ingredients,cells,and targets.The binding mode between the key ingredients and key targets was elucidated using molecular docking and molecular dy-namics simulation.Results Serum creatinine and blood urea nitrogen levels,tubulointerstitial inflamma-tion and tubular injury indexes,and renal fibrosis de-grees were significantly reduced by SWYST administra-tion.Quercetin,kaempferol,luteolin,baicalein and wogonin were considered as key components that main-ly acted through FOS and JUN of endothelial cells,neu-trophils and thick ascending limb cells to ameliorate CKD.Molecular docking and molecular dynamics sim-ulation revealed that quercetin stably occupied the cross pocket of FOS-JUN heterodimers to inhibit the binding between FOS-JUN heterodimers and DNA.Conclusion SWYST inhibits the binding of FOS-JUN heterodimers and DNA of endothelial cells,neutrophils and thick ascending limb cells to ameliorate CKD.