Over the past two decades, genome-wide association study(GWAS) has identified numerous genetic variants and loci associated with heritable diseases. With the gradual maturation and saturation of GWAS methodologies, transcriptome-wide association study(TWAS) offers a novel perspective by linkinggenetic phenotypes to gene expression levels. By integrating TWAS with other multi-omics analyses, researchers can gain a deeper understanding of heritable diseases. This article provides an overview of recent groundbreaking and representative TWAS methods and tools, analyzes their strengths and limitations, and discusses future trends in TWAS development.

ObjectiveTo observe the expression of miR-139/Notch1 axis and macrophage polarization in the homing changes of bone mesenchymal stem cells （BMSCs） in asthmatic rats， and to explore the possible mechanism of immune regulation by BMSCs during asthma. Methods30 male SD rats were randomly divided into three groups： normal control group， model control group and BMSCs implantation group， with 10 rats in each group. BMSCs labeled with CFSE were infused into the body of asthmatic rats through the tail vein， and the homing status of BMSCs in asthmatic lung tissue was detected by flow cytometry. Changes in the proportion of inflammatory cells in alveolar lavage fluid were detected by Wright-Giemsa Stain； the levels of macrophage polarization cytokines IFN⁃γ，IL-13，CD80 and CD206 in rat serum were detected by ELISA； the miR-139， Notch1， NOS2， Arg1 and CXCR4 in lung tissue were detected by RT-qPCR. ResultsCompared with the NC group， the expression of serum CD80 and IFN⁃γ in the MC group decreased， while the expression of IL-13 and CD206 increased （P<0.01）. The expression of miR⁃139 in lung tissue of MC group rats decreased， and the expression of macrophage polarization markers NOS2， Arg1， and homing marker CXCR4 genes increased （P<0.01）. Compared with the MC group， the expression of IFN-γ of rats in BMSCs group increased， while the expression of IL-13 and CD206 decreased （P<0.01）. The expression of miR⁃139， CXCR4， and SDF⁃1 mRNA in the lung tissue of rats of BMSCs group increased， while the expression of Notch1， NOS2， and Arg1 decreased （P<0.01）. Correlation analysis showed that CXCR4 was positively correlated with miR⁃139 （P<0.05）， while CXCR4 was negatively correlated with Notch1 （P<0.05）. SDF⁃1 and IFN⁃γ was a positively correlated （P<0.05）， while SDF⁃1 was negatively correlated with Arg1 and CD206 （P<0.05）. ConclusionThe miR⁃139/Notch1 axis can promote BMCs homing in asthmatic rats by affecting macrophage polarization in asthma.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

AIM To investigate the impact of varying dosages of Supplemented Wendan Decoction on the PI3K/Akt/FOXO1 glycolipid metabolic pathway in 3T3-L1 adipocytes.METHODS The CCK-8 assay was used to determine the concentration of Supplemented Wendan Decoction-medicated serum.The mature adipocytes differentiated from 3T3-L1 preadipocytes after induction were further divided into the blank control group,the model group,the rosiglitazone group(10 mg/L),and the Supplemented Wendan Decoction groups(5％,10％,and 20％),followed by the sample collections after 48 hours of treatment.Oil red O staining quantified lipid accumulation in 3T3-L1 adipocytes;extracellular glucose levels were measured using glucose oxidase(GOD)assay;RT-qPCR analyzed mRNA expressions of IRS-1,PI3K,Akt,GLUT4,IL-6,TNF-α and IL-1β;Western blot assessed protein expressions of INSR,IRS-1,PI3K-p85,Akt,FOXO1 and GLUT4.RESULTS No significant changes in cell viability(P＞0.05)were observed in 3T3-L1 preadipocytes exposed to serum containing supplemented Wendan Decoction at different concentrations for 24,48,or 72 hours.The 3T3-L1 preadipocytes held the capacity to differentiate into mature adipocytes within a 14-day induction period.Compared to the model group,all supplemented Wendan Decoction groups exhibited reduced lipid accumulation in adipocytes and downregulated mRNA expression of IRS-1,IL-6,TNF-α and IL-1β(P＜0.01);the low-dose group demonstrated increased mRNA expressions of PI3K and GLUT4(P＜0.05,P＜0.01),alongside elevated protein expressions of INSR,IRS-1,PI3K-p85,Akt and GLUT4(P＜0.05,P＜0.01);the medium-dose group showed enhanced GLUT4 mRNA expression,and upregulated protein expressions of INSR and FOXO1(P＜0.01).After 24 hours intervention,the high-dose Supplemented Wendan Decoction group exhibited increased glucose consumption in adipocytes(P＜0.01),and elevated protein expression of INSR,Akt and FOXO1(P＜0.05,P＜0.01).CONCLUSION Supplemented Wendan Decoction reduces lipid accumulation in adipocytes,regulates glucose and lipid metabolism,and promotes metabolic homeostasis through PI3K/Akt/FOXO1 signaling pathway.

AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.

To formulate an expert consensus on the principles of preoperative hair removal and provide scientific guidance for standardized removal of hair before surgical procedures so as to reduce the incidence of surgical site infections.METHODS Led by the Hospital Management Institute of National Health Commission of the People's Republic of China,this consensus was reached with the joint efforts from the expects of relevant fields such as surgeries,interventional therapies,nursing,and infection prevention and control.The consensus facilitates the classification and evaluation of literatures by following the evidence grade formulated by Oxford Evidence-based Medicine Center and focuses on the association of preoperative hair removal with surgical site infection,it reaches the evidence grade of expert consensus and recommendation intensity by integrating with discussions on meetings and clinical experience of the expects from relevant fields.RESULTS A total of 6 items of consensus were reached by summarizing the latest evidence on the aspects including the indications for preoperative hair removal,tools,range,timing and places.CONCLUSION The consensus,to some extent,make supplements to and complete the exiting regulations and standards.It provides guidance for the medical institutions to carry out the preoperative hair removal.

The health-associated infection in the department of stomatology is an important issue for global public health and is closely associated with the medical quality and the health of health care workers and patients.Al-though certain progress has been achieved in the prevention and control of the health-associated infections in de-partment of stomatology,considering the diversity of influencing factors,the joint effort of the global public health institutions,medical institutions and scientific research institutions remains necessary for the effective con-trol of the hospital-associated infections.The study focuses on the current status of hospital-associated infections in department of stomatology,risk factors and progress of research on prevention and control strategies in recent years so as to formulate more effective prevention and control strategies and provide reference for better prevention and control of the infections and protection of public heath.

Objective:To investigate the prevalence of scoliosis among middle school students through school-based screening, and to analyze the correlations between axial trunk rotation (ATR) angle and Cobb angle among those with scoliosis considering gender, age, body mass index (BMI) and curve type, so as to provide references for early prevention and treatment.Methods:Random cluster sampling was used to enroll 9501 middle school students from 11 schools in the Hanyang District of Wuhan. They underwent a six-step scoliosis screening with a forward bending test, scoliometer measurements, and confirmatory X-ray examinations. The students exhibiting scoliosis were stratified by gender, age, BMI, and curve type. The correlation between ATR angle and Cobb angle was subsequently analyzed within each subgroup.Results:Scoliosis was observed in 358 of the students (150 boys and 208 girls), a positive screening rate of 3.8%. Moreover, the positive screening rate was significantly higher among the girls (4.8%) than among the boys (2.9%). There was a moderate Spearman correlation ( r=0.69) between the ATR and Cobb angles in 358 of the students with scoliosis, both the boys ( r=0.69) and the girls ( r=0.70). When it came to age, there was a positive correlation between the ATR and Cobb angles for students aged 13 to 16 irrespective of their BMI classification, except that there was no significant correlation between the two angles for those classified as obese. In terms of Peking Union Medical College (PUMC) classification, there was a positive correlation for those in types Ia, Ib, Ic, IIa, IIb, IIc and IId, but there was no significant correlation between the two angles for those in type III. Conclusions:Scoliosis is prevalent among the group of adolescents studied. Society, schools and parents need to prioritize prevention, heightened awareness, timely diagnosis and early treatment, particularly for girls. The ATR angle may serve as a practical surrogate for estimating Cobb angle severity in scoliosis screening for single-curve or double-curve cases. However, ATR measurements may yield false-negative results among those with triple-curve deformities or the obese, underscoring the need for supplementary diagnostic methods among such populations.

Aim To investigate the promotive effects and mechanisms of the combined use of brucine(Bru)and lumbrokinase(LK),active ingredient derived from Longma formula,in promoting chondrocyte proliferation via the Wnt/β-catenin signaling pathway.Methods The extracted primary rat chondrocytes were divided in-to the following groups:Control group,Bru,LK alone group,and Bro+LK combination group.The optimal drug concentration and intervention time were deter-mined using CCK-8 assay,followed by cell proliferation validation through EdU and phalloidin staining.The expression levels of collagen Ⅱ,aggrecan and SRY-re-lated high-mobility group box gene 9(SOX9)in chon-drocytes following intervention with the combination of Bru and LK were detected by Western blotting.Addi-tionally,the regulatory effects of these proteins on the Wnt/β-catenin signaling pathway were also investiga-ted.Results The optimal combination concentration of Longma formula active ingredients(Bru 0.025 mg·L-1+LK 5 mg·L-1)significantly enhanced chondro-cyte viability compared to control,Bru,or LK alone at 48 h.This combination increased the S-phase ratio,promoted the aggregation of intracellular actin fila-ments,and upregulated the expression of collagen Ⅱ and aggrecan.Furthermore,it activated the Wnt/β-catenin pathway,leading to increased SOX9 expres-sion.Conclusions The optimal combination of Bru and LK(Bru 0.025 mg·L-1+LK 5 mg·L-1)de-rived from Longma formula significantly maintains chondrocyte phenotype and promotes cellular prolifera-tion through the activation of the Wnt/β-catenin signa-ling pathway,which subsequently upregulates the downstream target SOX9.

Objective To establish the reference interval of urine Alzheimer-associated neuronal thread protein(AD7c-NTP)for healthy adults in Mianyang area using the indirect method.Methods The detection results of urine AD7c-NTP from 5 093 healthy in-dividuals were collected from the information management database of Medical Laboratory Department of Sichuan Science City Hospital from March 2017 to March 2022.Skewness-kurtosis and Kolmogorov-Smirnov tests were used to determine whether the data followed a normal distribution.After removing outliers using the Box Plots method,the enrolled subjects were grouped by gender and age.The Mann-Whitney U or Kruska-Wallis H tests were used to analyze the between-group differences of urine AD7c-NTP in healthy individu-als with different genders and ages.The adjacent age groups without statistically significant difference(P＞0.05)were combined,and the indirect method(non-parametric test method)was used to calculate the reference intervals for different gender and age groups.Results Skewness-kurtosis and Kolmogorov-Smirnov tests showed that the data followed a non-normal distribution.After removing 293 outliers using the Box Plots method,a total of 4 800 subjects,including 3 199 males and 1 601 females,were enrolled.The enrolled subjects were grouped by gender and age,and the non-parametric test method were used to establish the reference intervals of urine AD7c-NTP in healthy populations with different genders.The Mann-Whitney U test confirmed that urine AD7c-NTP levels existed gen-der differences(Z=14.09,P＜0.01),and the reference intervals for males and females were≤1.10 ng/mL and≤1.40 ng/mL,re-spectively.There were also statistical differences in urine AD7c-NTP levels among different age groups of the same gender.After combi-ning adjacent age groups without statistically significant difference(P＞0.05),the reference intervals of urine AD7c-NTP in healthy populations with different genders and ages were established by the non-parametric test method,which were≤1.00 ng/mL for male 20-39 years old group,≤1.10 ng/mL for male 40-79 years old group,≤1.60 ng/mL for male≥80 years old group,≤1.30 ng/mL for female 20-69 years old group,and≤1.60 ng/mL for female≥70 years old group,respectively.The established reference intervals of urine AD7c-NTP were further verified by healthy individuals,and the results met the standards.Conclusion The reference intervals of urine AD7c-NTP in healthy populations with different genders and ages in Mianyang area are established successfully using the indi-rect method,which may help to predict the risk of Alzheimer's disease in clinical practice and provide support for the diagnosis and treatment of the disease.