The gut microbiota is regarded as the "eighth organ" of the human body and plays a critical regulatory role in the occurrence and progression of various diseases. Ulcerative colitis （UC） is a chronic inflammatory bowel disease with a complex etiology and a tendency toward recurrent episodes. In recent years， studies have shown that gut microbiota dysbiosis plays a key role in its pathological processes. Meanwhile， an increasing number of studies have demonstrated that imbalances in the gut microbiota and abnormalities in its metabolites are closely associated with the development of atrial fibrillation （AF）. Although UC and AF belong to diseases of the digestive system and cardiovascular system， respectively， both exhibit systemic inflammatory characteristics and are often accompanied by gut microbiota dysregulation and abnormal metabolic products. However， systematic investigations into the mechanisms by which gut microbiota-derived metabolites act in these two diseases remain limited. Based on this， the present study adopts literature review and theoretical analysis methods， taking the "gut microbiota-gut-heart" axis as the entry point， to systematically summarize the signaling networks of three key classes of metabolites， i.e.， short-chain fatty acids （SCFAs）， bile acids （BAs）， and trimethylamine N-oxide （TMAO）， in the comorbidity mechanism of UC and AF. The findings indicate that these metabolites may activate key inflammatory pathways， such as NF-κB and NLRP3， thereby synergistically mediating intestinal barrier dysfunction and systemic inflammation and constructing a potential comorbidity network. On this basis， potential intervention strategies for the treatment of UC-AF comorbidity， including probiotic intervention and fecal microbiota transplantation， are further discussed. This study aims to provide new theoretical evidence and research perspectives for prevention and treatment strategies of cross-system diseases.

ObjectiveTo meet the requirements of Good Laboratory Practice (GLP), Sichuan Institute for Drug Control (Sichuan Testing Center of Medical Devices, Sichuan Institute of Musk Deer Breeding) Safety Evaluation Center (hereinafter referred to as the center) conducts a targeted validation study to address the lack of supporting data for the specified validity periods of disinfectants, feed, and bedding during their use in laboratory animal facilities. The study summarizes practical experience in material management and provides a scientific basis for the standardized management of materials in the facility. MethodsUnder the routine operating conditions of the laboratory animal facility at the center, the disinfection efficacy of four commonly used disinfectants was confirmed using the quantitative suspension kill test, with the disinfectants tested on day 0, day 7, and at 1, 2, and 3 months after preparation and after opening; for laboratory animal feed, nutritional components, chemical contaminants, and microbial indicators were tested 7 days after opening; microbial indicators of laboratory animal corncob bedding were tested after storage for 7 days following opening and sterilization. ResultsUnder the GLP management system of our center, the disinfection efficacy log reduction values of prepared 0.04% sodium hypochlorite solution, 0.1% didecyldimethylammonium bromide solution, and 0.2% benzalkonium bromide solution were all ≥5 on days 0 and 7. The log reduction values of opened 75% ethanol were ≥5 on days 0 and 7 and at 1, 2, and 3 months after opening. All disinfectants were therefore qualified and effective. Compared with the factory inspection data, the nutritional components, chemical contaminants, and microbial indicators of SPF-grade rat and mouse maintenance feed, SPF-grade guinea pig growth and reproduction feed, conventional-grade guinea pig growth and reproduction feed, rabbit maintenance feed, and dog maintenance feed showed a slight overall downward trend but no significant differences 7 days after opening. The microbial indicators of specialized corncob bedding for SPF-grade laboratory animals stored for 7 days after opening and conventional-grade corncob bedding stored for 7 days after sterilization showed no significant fluctuations. All indicators still met the relevant requirements of GB/T 14924.2—2001 Laboratory Animals—Hygienic Standard for Formula Feeds and GB 14924.3—2010 Laboratory Animals—Nutrients for Formula Feeds. ConclusionBased on practical experience in managing the center's GLP laboratory animal facility and validation of core quality indicators, the specified validity periods of disinfectants, feed, and bedding meet the quality requirements, providing a practical reference for the standardized management of laboratory animal facilities.

Objective:To screen for microRNAs(miRNAs)highly expressed in the serum exosomes(Exo)of esophageal squamous cell carcinoma(ESCC)patients and analyze their relationship with the clinicopathological characteristics of the patients,and to explore the potential of Exo-derived miRNAs as clinical auxiliary diagnostic markers for ESCC.Methods:Serum and relevant clinical data of 50 healthy subjects and 45 newly diagnosed ESCC patients admitted to the Fourth Hospital of Hebei Medical University between December 2021 and June 2023 were collected,serving as the control group and the ESCC group respectively.The Gene Expression Omnibus(GEO)database and qPCR were used to screen and identify the candidate miRNA for increased expression in the serum of ESCC patients-miR-1246.The diagnostic efficacy of serum miR-1246 for ESCC was analyzed by the receiver operating characteristic curve.The relationship between miR-1246 and the clinical feature progression of ESCC patients was analyzed by Logistic regression,and the relationship between miR-1246 and the clinicopathological characteristics of ESCC patients was analyzed by the χ2 test.Exosomes in the serum of the subjects were isolated,purified and characterized for verification.The expression of miR-1246 in Exo was detected by qPCR.ESCC KYSE150 and KYSE30 cells were routinely cultured.mimics-NC and miR-1246 mimics were transfected respectively into KYSE150 cells using Lipofectamine 2000.Inhibitor-NC and miR-1246 inhibitor were transfected into KYSE30 cells,which were respectively denoted as the minics-NC,miR-1246 mimics,inhibitor-NC and miR-1246-inhibitor groups.KYSE150 and KYSE30 cells were treated with Exo derived from KYSE150 cells in the mimics-NC and miR-1246 mimics groups.The proliferation,migration and invasion abilities of cells in each group were detected by the CCK-8 assay,scratch wound healing assay and Transwell chamber assay respectively.The expressions of Exo markers,epithelial-mesenchymal transition-related proteins,TET family methylcytosine dioxygenase 2(TET2)and cell adhesion molecule 1(CADM1)proteins in each group of cells were detected by WB assay.The targeting binding relationship between miR-1246 and TET2 and CADM1 was verified by the dual-luciferase reporter gene assay.Results:Bioinformatics screening showed that the miRNA with the most significant differential expression in the serum of ESCC patients was miR-1246.The serum Exo extracted from the patients conformed to the typical Exo characteristics.The expression level of serum Exo-miR-1246 in ESCC patients at stages Ⅰ-Ⅱ was significantly higher than that in healthy subjects(P<0.01);the level of serum Exo-miR-1246 in ESCC patients at stages Ⅲ-Ⅳ was significantly higher than that in patients at stages Ⅰ-Ⅱ(P<0.01).ROC curve analysis showed that Exo-miR-1246 in serum had a high value for auxiliary differential diagnosis of ESCC(P<0.05),and the auxiliary diagnostic efficacy of Exo-miR-1246 for the clinical progression of ESCC patients was higher than that of CEA and SCC-Ag(P<0.05).The combined detection of the three could further improve the efficacy of auxiliary diagnosis of patient staging(P<0.01).Exo-miR-1246 might be an independent risk factor for the clinical progression of ESCC patients(P<0.05).The expression level of serum Exo-miR-1246 was associated with the T-stage,N-stage and clinical stage of ESCC(P<0.01).Overexpression of miR-1246 could promote the proliferation,migration,invasion,epithelial-mesenchymal transition and inhibit apoptosis of ESCC cells,while inhibition of miR-1246 had the opposite effect.Database data analysis found that TET2 and CADM1 were the target genes of miR-1246.The dual-luciferase reporter gene assay confirmed that miR-1246 could directly bind to TET2 and CADM1 mRNA and inhibit their expressions(P<0.01).Treatment of KYSE150 and KYSE30 cells with Exo derived from cells overexpressing miR-1246 had the same effect as overexpressing miR-1246 in these cells.Conclusion:Exo-derived miR-1246 has the potential to be a clinical auxiliary diagnostic marker for ESCC.It may affect the occurrence and development of ESCC by regulating the expression levels of TET2 and CADM1.

Objective:To investigate the impact of preoperative sarcopenia on chronic postsurgical pain(CPSP)after cardiac surgery in elderly patients.Methods:Elderly patients undergoing elective open-chest cardiac surgery at the Affiliated Hospital of Xuzhou Medical University from September 2022 to May 2024 were collected.According to the updated diagnostic criteria and revised by the Asian Working Group for Sarcopenia(AWGS2019)in 2019, patients were classified into sarcopenia and non-sarcopenia groups Elderly patients were divided into two groups based on the occurrence of chronic pain at 3 months postoperatively: CPSP group and non-CPSP group.Indicators with statistically significant differences in univariate regression analysis were included in multifactorial regression to analyze the influencing factors of chronic pain after cardiac surgery in elderly patients.The receiver operating characteristic(ROC)curve was plotted and the area under the curve(AUC)was calculated to compare the predictive efficacy of sarcopenia, commonly used clinical pain assessment tools(gender+ acute postoperative pain), and(gender+ acute postoperative pain+ sarcopenia)in predicting CPSP after cardiac surgery in elderly patients.Results:The study ultimately included 379 patients, consisting of 238 males(62.8%), with an average age of(66.6 ± 5.3)years.Among them, 83 patients had sarcopenia, and 119 patients developed CPSP.Univariate regression analysis showed that gender, history of atrial fibrillation, acute postoperative pain, American Society of Anesthesiologists(ASA)Physical Status Classification System, New York Heart Association(NYHA)Classification of Cardia Function, sarcopenia, and duration of extracorporeal circulation were associated with the occurrence of CPSP after cardiac surgery in elderly patients.However, after adjusting for all possible confounders, multifactorial regression analysis showed that gender, acute postoperative pain, and sarcopenia were independent risk factors for CPSP after cardiac surgery in elderly patients(all P<0.05), with sarcopenia patients having a 2.913-fold risk of developing CPSP compared with non-sarcopenia patients.The AUCs of the ROC curves for commonly used clinical perioperative pain assessment tools and those with the addition of sarcopenia determination were 0.731 and 0.802, respectively. Conclusions:Preoperative sarcopenia is an independent risk factor for the development of chronic pain after cardiac surgery in elderly patients, and the inclusion of sarcopenia determination in commonly used clinical pain assessment tools can significantly improve the predictive efficacy for chronic pain.

Objective To investigate the correlation between serum amyloid A(SAA)level and disease se-verity in children infected with severe acute respiratory syndrome coronavirus 2(COVID-19).Methods A to-tal of 116 children infected with COVID-19 admitted to the Department of Pediatrics of the hospital from De-cember 2022 to April 2023 were included and divided into asymptomatic/mild group and moderate/severe group according to the severity of the disease.In addition,65 healthy children who received health examination during the same period were selected as the control group.Serum SAA levels in children in acute stage and convalescent stage were detected by enzyme-linked immunosorbent assay.Results Compared with the control group,the serum SAA level in the children infected with COVID-19 was significantly increased in the acute stage(P＜0.05).The area under the receiver operating characteristic(ROC)curve(AUC)of SAA levels in the acute stage for diagnosing children with COVID-19 infection was 0.926(95％CI:0.886-0.966).In SARS-CoV-2 infected children,the SAA levels in the acute stage in the asymptomatic/mild group and the moderate/severe group were 2.71(1.29-10.86)mg/L and 37.78(18.58-92.62)mg/L,the differences were statistically significant between the two groups(Z=5.782,P＜0.001).In addition,serum SAA was pos-itively correlated with severity of SARS-CoV-2 by Spearman analysis(r=0.657,P＜0.001).Serum SAA lev-els were also significantly positively correlated with C-reactive protein(CRP),immunoglobulin(Ig)M,IgG,IgA and neutralizing antibody(NAb)in acute stage(P＜0.05).The AUC of serum SAA level in acute stage for diagnosing the moderate/severe children with SARS-CoV-2 was 0.889(95％CI:0.842-0.955),which was higher than that of CRP(P＜0.05).Compared with serum antibodies(IgM,IgG,IgA and NAb),the rate of serum SAA positive(≥5.55 mg/L)in children in acute stage was significantly higher(P＜0.05).The positive rate of serum SAA in convalescent children was significantly lower than that of serum antibody(P＜0.05).Conclusion Elevated serum SAA in acute phase is associated with increased risk of SARS-CoV-2 infec-tion and disease severity in children.Serum SAA is promising as a good biomarker for monitoring SARS-CoV-2 infection and severity in children.

Objective To identify genes regulated by ME2 and to explore their roles as well as underlying mecha-nisms in liver cancer progression.Methods RNA-seq data of siME2-transfected cells were subjected to differential expression analysis,clustering,GO and KEGG enrichment analyses.The mRNA level of the potential target gene SHCBP1 was measured by quantitative real-time PCR(qPCR)following ME2 knockdown or overexpression in HepG2 cells.The effect of ME2 and SHCBP1 on the downstream pathway was examined by Western blot.Cell pro-liferation,wound healing,and colony formation assays were conducted to evaluate SHCBP1's role in liver cancer cell proliferation and migration.Survival analysis of the TCGA-LIHC cohort was performed to determine the prog-nostic value of SHCBP1 in liver cancer patients.Results Differentially expressed genes in siME2-transfected cells were significantly enriched in biological processes including the PI3K-Akt signaling pathway,cell cycle,and serine phosphorylation.In HepG2 cells,ME2 knockdown led to a reduction in SHCBP1 mRNA level,whereas ME2 over-expression resulted in enhanced SHCBP1 mRNA level,demonstrating a positive correlation between ME2 and SHCBP1 expression.Western blot analysis revealed that ME2 enhanced PI3K-Akt signaling pathway activation through SHCBP1.qPCR results confirmed that SHCBP1 was significantly over-expressed in liver cancer cells and promoted both proliferation and migration,contributing to poor prognosis in liver cancer patients.Conclusions ME2 promotes liver cancer progression by regulating SHCBP1 to activate the PI3K-Akt signaling pathway,presen-ting a novel therapeutic target for liver cancer treatment.

Objective To investigate the value of constructing a risk prediction model of brain metastasis in lung cancer based on clinical-CT imaging.Methods The clinical and CT imaging data of 208 patients with lung cancer confirmed by surgical pathology or puncture biopsy were analyzed retrospectively,including 98 patients in the metastasis group and 110 patients in the non-metastasis group.Univariable and binary logistic regression analyses were performed between the two groups,and the clinical,CT imaging,and clinical-CT imaging models were constructed according to the selected independent risk factors.Prediction model performance was eval-uated with receiver operating characteristic(ROC)curve,calibration curve and decision curve analysis(DCA).Results Multivariate analysis showed that T stage,pathological type,radiotherapy and chemotherapy,surgery,long diameter(LD),short diameter(SD),minimum CT value(CTmin)were the independent risk factors for predicting brain metastasis in lung cancer(P<0.05).The area under the curve(AUC)of clinical,CT imaging and clinical-CT imaging models were 0.925,0.764,0.941,respectively.DeLong test analysis showed that the AUC of clinical-CT imaging model,clinical model and CT imaging model was statistical difference(Z=2.093,5.777,all P<0.05).The calibration curve suggested a good fit of the clinical-CT imaging model.The DCA suggested that the clinical-CT imaging model demonstrates good clinical benefits.Conclusion The clinical-CT imaging model can effectively predict the occurrence of brain metastasis in lung cancer,which is helpful to guide the development of accurate diagnosis and treatment plan.

BACKGROUND:Mammary stem cells are vital for the development and homeostasis of mammary gland tissue.The occurrence of breast cancer has a close relationship with the mammary stem cells.Recent studies have shown that Hopx,as an important transcriptional regulator of morphogenesis and cell differentiation,has been confirmed to be expressed in a variety of adult stem cells such as nerves,intestines,hair follicles and lungs.However,its role in mammary stem cells has not been reported so far.OBJECTIVE:To investigate whether Hopx expression marks mammary stem cells.METHODS:(1) Female Hopx-LacZ transgenic mice aged 8 weeks were selected to detect the background expression of Hopx in breast tissue by β-galactosidase staining.(2) Female wild-type mice at 4,6,and 8 weeks of age and 14.5 days of gestation were selected for whole-tissue magenta staining and K14 and K8 immunofluorescence staining,respectively.(3) Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks and 17.5 days of gestation were selected and stained with breast β-galactosidase.(4) The 4-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times),and breast β-galactosidase staining was performed 4 weeks after injection.The 8-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times),and breast β-galactosidase staining was performed 4 and 10 weeks after the last injection.(5) Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen (once every other day,three times).Hopx-CreERT2;Rosa26LacZ transgenic mice were pregnant 2 weeks after injection.The mammary tissue of mice at 17.5 days of the first pregnancy and 17.5 days of the third pregnancy was stained with β-galactosidase.RESULTS AND CONCLUSION:(1) The results of β-galactosidase staining showed that the mammary ducts of Hopx-LacZ transgenic mice at 8 weeks of age did contain Hopx-positive cells and were located in the basal epithelia,with a small number.(2) Whole-mount staining of mammary glands and immunofluorescence staining results exhibited that the mammary glands of mice had different characteristics with corresponding developmental stages such as puberty,maturity,and pregnancy,and underwent a series of complex epithelial remodeling processes.(3) The results of β-galactosylase staining showed that Hopx-labeled positive cells in the mammary duct of Hopx-CreERT2;Rosa26LacZ transgenic mice at 17.5 days of gestation increased compared with female Hopx-CreERT2;Rosa26LacZ transgenic mice at 8 weeks of age.(4) The results of β-galactosylase staining showed that the Hopx-labeled positive cells in the mammary glands of 4-and 8-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice after tamoxifen injection were located in the basal epithelium with a small number.(5) The results of β-galactosidase staining showed that Hopx-labeled positive cells in the mammary glands of mice at 17.5 days of the first and third gestation were located in the basal epithelia around the alveoli,and the number of Hopx-labeled positive cells at 17.5 days of the third gestation was more.(6) In conclusion,Hopx reporter-marked basal epithelial cells belong to dormant mammary stem cells,which are responsible for the growth of the mammary glands during pregnancy and contribute to acinar formation.

Eosinophilic gastroenteritis(EG)is a rare disease characterized by abnormal infiltration of eosinophils(Eos)in gastrointestinal tissues.Due to the unclear pathogenesis of EG and the lack of effective therapeutic drugs,research on its novel mechanisms,targets and drugs is critical.This article starts by outlining the research progress in the pathogenesis of EG,involving IgE mediated typeⅠimmediate allergic reactions and T helper 2 cell(Th2)mediated delayed allergic reactions.Then,the related targets of EG are summarized,including Th2 cytokines and factors regulating Eos function,but there has been no breakthrough in the treatment of these targets.Finally,the therapeutic drugs for EG are reviewed,such as glucocorticoids,antiallergic drugs and biologics.The advantages and disadvantages of various drugs are also described.However,these drugs cannot meet the current demands of clinical treatment and there is an urgent need to develop novel therapeutic drugs.It is believed that multi-target therapy is an ideal treatment for EG,and that traditional Chinese medicine and natural products should be the priorities of research and development for EG therapeutic drugs in the future.This review is expected to provide new ideas for the clinical treatment strategies and drug development of EG.