ObjectiveTo construct a risk prediction model for frequent acute exacerbations of chronic obstructive pulmonary disease （COPD） under disease-syndrome combination， thus providing decision support for precise clinical intervention. MethodsA total of 2 029 patients with acute exacerbations of COPD admitted to the First Affiliated Hospital of Anhui University of Chinese Medicine from January 2020 to August 2024 were retrospectively included. These patients were classified into groups of frequent acute exacerbations （≥2 times/year） and infrequent acute exacerbations （<2 times/year） according to the hospitalization times per year. Risk factors were screened by LASSO regression combined with logistic regression， and a nomogram model was constructed. The model performance was assessed based on the area under the curve （AUC）， calibration curves， and decision curve analysis （DCA）. ResultsThe differences in baseline characteristics between the frequent acute exacerbations group （1 196 cases） and infrequent acute exacerbations group （833 cases） were not statistically significant. LASSO regression combined with multivariate logistic regression screened the following independent risk factors： body mass index （BMI）， hospitalization days， number of smoking years， place of residence， use of noninvasive ventilators， oxygen-demanding therapy， liver cirrhosis， use of systemic glucocorticosteroids， and traditional Chinese medicine syndrome （phlegm and stasis obstructing the lung）. The nomogram model showed good discrimination and calibration in both the training set （AUC=0.748） and validation set （AUC=0.774）. ConclusionThe risk prediction model for frequent acute exacerbations of COPD， integrating traditional Chinese medicine syndrome， constructed in this study has high accuracy. It can provide a scientific basis for early clinical identification of high-risk patients and individualized intervention.

Objective:To explore the ultrasound changes,clinical efficacy and safety of ultrasound-guided compound betamethasone injection in the treatment of first metatarsophalangeal joint(MTP1)in acute gouty arthritis.Methods:This study was a prospective randomized controlled trial,71 patients diagnosed with MTP1 in acute gouty arthritis in Weifang People's Hospital from June 2022 to June 2024 were selected,they were divided into puncture treatment group of 36 cases and oral medication group of 35 cases according to the random number table method.The puncture treatment group was treated with ultrasound-guided compound betamethasone injection,and the oral medication group was treated with etocoxib combined with colchicine.Visual Analogue Scale(VAS),inflammatory indicators[erythrocyte sedimentation rate(ESR),C-reactive protein(CRP)],and semi-quantitative scores of musculoskeletal ultrasound of the two groups were compared,and the medication safety of the two groups was observed.Result:1,3 and 7 days after treatment,VAS score in the puncture treatment group decreased significantly compared with that in the oral medication group(P＜0.05).7 days after treatment,ESR and CRP in the puncture treatment group decreased more significantly,and semi-quantitative scores of musculoskeletal ultrasound showed that the improvement of joint effusion,synovial hyperplasia and blood flow signal was better than that in the oral medication group(P＜0.05).All 36 cases of puncture were successful at the first attempt without complications.Conclusion:Ultrasound-guided compound betamethasone injection in the treatment of MTP1 in acute gouty arthritis has a high success rate,it can rapidly reduce inflammation,relieve pain,restore joint function,and is safe to operate,it can be used as an effective treatment option.

Objective:To investigate the reparative effects and underlying mechanisms of human umbilical cord mesenchymal stem cells (hUCMSCs) in conjunction with hyperbaric oxygen (HBO) on ovarian dysfunction in mice exposed to simulated high-altitude conditions.Methods:A total of 64 six-week-old female C57BL/6 mice were randomly allocated according to random number table, assigned to control group, model group, hUCMSCs group and hUCMSCs+HBO group, with 16 mice in each group. Mice in model group, hUCMSCs group and hUCMSCs+HBO group were placed in a hypobaric oxygen chamber to simulate a high-altitude environment at an elevation of 6 500 m for 15 d, thereby establishing a model of ovarian dysfunction. Beginning on the first day following model establishment, the control and model groups received intravenous injections of 0.2 mL saline via the tail vein. In contrast, the hUCMSCs group and the hUCMSCs+HBO group received 0.2 mL hUCMSCs (1×10 6 cells) per mouse through tail vein injection, administered once a week for three weeks, with continuous intervention lasting 15 d. Furthermore, the hUCMSCs+HBO group were subjected to daily hyperbaric oxygen treatment. The recorded variables included general health status, body weight, estrous cycle changes, serum hormone levels, bilateral ovarian wet weight, ovarian index, follicular development assessment, pathological alterations of ovarian tissue, ultrastructural changes, and the expression levels of transforming growth factor beta 1 (TGF-β1) and Smad family member 3 (Smad3) detected by Western blotting in ovarian tissue. Additionally, litter size and offspring body weight were measured across all groups to evaluate the reproductive capacity of the mice. Results:1) Compared with the hUCMSCs group, the hUCMSCs+HBO group exhibited no statistically significant differences in estrous cycle, body weight, bilateral ovarian wet weight, or ovarian index (all P>0.05). Furthermore, serum levels of anti-Müllerian hormone (AMH), estradiol, progesterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH), as well as counts of various stages of follicles in hUCMSCs+HBO group, did not demonstrate statistically significant differences compared with hUCMSCs group (all P>0.05). Notably, in the hUCMSCs+HBO group, the integrity of the nuclear membrane in granulosa cells of the ovarian tissue was preserved, with only mild swelling observed in individual mitochondria, while no expansion of the rough endoplasmic reticulum or swelling of the Golgi apparatus were noted. Additionally, expression levels of TGF-β1 and Smad3 proteins in the hUCMSCs+HBO group were significantly elevated compared with the hUCMSCs group ( P=0.010, P<0.001). The conditions of the offspring and litter size showed no statistically significant differences between the two groups (all P>0.05). 2)Compared with control group, the hUCMSCs+HBO group had slightly lower values for body weight, bilateral ovarian wet weight, ovarian index, serum levels of AMH, estradiol, and progesterone, while had slightly higher serum levels of FSH and LH without statistically significant differences (all P>0.05). Importantly, the ultrastructural characteristics of granulosa cells in the ovarian tissue of the hUCMSCs+HBO group closely resembled those of control group, displaying intact structures of the nuclear membrane, mitochondria, rough endoplasmic reticulum, and Golgi apparatus. There was no significant difference in TGF-β1 protein level between hUCMSCs+HBO group and control group ( P=0.253), but Smad3 protein level in hUCMSCs+HBO group was higher than that in control group ( P<0.001). No statistically significant differences were detected in offspring body weight, litter size, or behavioral tendencies (all P>0.05). Conclusion:Both hUCMSCs and the combination of hUCMSCs with HBO intervention demonstrated a safe and effective promotion of functional repair in damaged ovarian tissue under hypoxic conditions. Notably, the combination treatment of hUCMSCs with HBO exhibited a synergistic effect compared with hUCMSCs alone. The potential mechanism underlying this enhanced functional repair may involve the upregulation of the TGF-β1/Smad3 signaling pathway, which could ultimately improve fertility outcomes in mice subjected to hypoxic environments.

Objective Prepubertal mice are administered subcutaneously with letrozole sustained-release pellets behind the neck and treated with a high-fat diet to establish a mouse model of polycystic ovary syndrome(PCOS).The liver transcriptomes of the model mice are compared with those of the placebo control mice to investigate the underlying mechanisms of liver involvement in the pathogenesis of PCOS.Methods A customized 2 mg dose of letrozole sustained-release pellets with a 40-day release period was used.The control placebo and letrozole pellets were implanted subcutaneously in the dorsal cervical region of 3-4-week-old C57BL/6J mice(8 mice per group)to establish the control group and letrozole-induced PCOS model group.Both groups were treated with a high-fat diet starting the day after administration.The modeling period lasted for 5 weeks,during which body weight and 24-hour food intake were monitored in each group every week.When samples were collected,liver weight was recorded.Pathological changes in ovarian and hepatic tissues were examined by hematoxylin-eosin(HE)staining,while hepatic lipid deposition was observed by Oil Red O staining.The extent of macrophage infiltration in the liver was evaluated via F4/80 immunohistochemical staining,and hepatic fibrosis levels were observed by Masson's trichrome staining.Transcriptomic sequencing was performed to analyze differentially expressed genes(DEGs)in liver tissues between the control and model groups,followed by enrichment analysis of significant DEGs.Quantitative real-time fluorescent quantitative PCR(qPCR)was subsequently used to validate the expression of significant DEGs in liver tissues of both groups.Results Compared with the control group,the model group which received subcutaneous letrozole sustained-release pellets combined with a high-fat diet exhibited significantly increased body weight(P＜0.001),prominent polycystic ovarian morphology,and significantly decreased liver-to-body weight ratio(P＜0.05).However,no significant changes were observed in absolute liver weight(P＞0.05),hepatic histomorphology,or lipid deposition.Transcriptome sequencing identified 119 upregulated and 217 downregulated DEGs in the liver tissues of letrozole-treated mice,which were predominantly enriched in pathways related to cholesterol and steroid biosynthesis,steroid hormone metabolism,and inflammatory responses.qPCR validation demonstrated that mRNA expression of HSD3B2 and HMGCR was significantly upregulated in liver(P＜0.01),while mRNA expression of IL4,CCL2 and COL1A1 was downregulated(P＜0.05)in the model group compared with the control group.However,Masson's trichrome staining and F4/80 immunohistochemical analysis showed no significant changes in hepatic fibrosis or macrophage infiltration.Conclusion Subcutaneous administration of letrozole sustained-release pellets combined with a high-fat diet successfully establishes a mouse model of PCOS.The model mice exhibited significant changes in hepatic gene expression.Liver may contribute to PCOS pathogenesis through regulating cholesterol and steroid metabolism.

In recent years,the incidence and mortality of heatstroke have been increasing annually alongside global warming,with a marked rise in cases exhibiting atypical symptoms.To address the increasingly complex challenges in heatstroke prevention and treatment,Heatstroke Prevention and Treatment Research Center of Chinese PLA,Expert Group of Heatstroke Prevention and Treatment of Chinese PLA,and Chinese PLA Professional Committee of Critical Care Medicine have jointly developed this guideline(2025 edition).Utilizing the Grading of Recommendations Assessment,Development and Evaluation(GRADE)system,Appraisal of Guidelines for Research and Evaluation(AGREE)criteria,and Reporting Items for Practice Guidelines in Healthcare(RIGHT)standards,and based on the 2015 draft"Expert Consensus on the Standardized Diagnosis and Treatment of Heatstroke"and the 2019"Chinese Expert Consensus on the Diagnosis and Treatment of Heatstroke",this guideline has been crafted.This guideline provides 25 evidence-based recommendations to guide the prevention,treatment and research of heatstroke,which thoroughly covers 8 critical domains:clinical classification,pathophysiological mechanisms,clinical manifestations,diagnostic criteria,differential diagnosis,treatment protocols,rehabilitation and return to work,and prevention.

Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25％hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25％HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90％,specificity of 100％,positive prediction rate of 100％,negative prediction rate of 83％,false positive rate of 0％,false negative rate of 17％and accuracy of 93％.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.

Aim To explore the mechanism of Lycium barbarum glycopeptide(LbGP)improving aging in rat primary ovarian granulosa cells.Methods This study divided the cells into a normal group,a DOX group,and four different LbGP concentration treatment groups post-DOX intervention.Results Cell proliferation was assessed using CCK-8,EDU,and Ki67 assays,while aging markers and mitochondrial function-related fac-tors were detected using immunofluorescence and West-ern blotting.The results showed that,compared to the DOX group,LbGP treatment significantly increased cell viability(P＜0.05)and promoted proliferation(P＜0.05).Post LbGP treatment,the β-galactosidase-posi-tive area in cells was significantly reduced compared to the DOX group(P＜0.05).Immunofluorescence re-sults indicated that,compared to the DOX group,levels of p21 and γH2AX significantly decreased(P＜0.05),while pRB increased(P＜0.05)after LbGP treatment.Western blot results showed that,compared to the DOX group,the aging phenotype proteins p21 and p53 significantly decreased(P＜0.05),and pRB notably increased(P＜0.05)in the LbGP treatment group.The release of cytC into the cytoplasm and the activated caspase-9 significantly decreased(P＜0.05);levels of CAMKK2,pAMPK,and mitochondrial calcium homeostasis regulator MCU increased(P＜0.05);nuclear energy metabolism-related proteins SirT1,PGC1α/β and ATP5A1 significantly increased(P＜0.05);compared to the DOX group,ROS levels significantly decreased after LbGP treatment(P＜0.05).Conclusions The results suggest that LbGP can ameliorate DOX-induced aging in rat primary ovar-ian granulosa cells,potentially through the upregulation of the CAMKKβ/AMPK signaling pathway,thereby im-proving mitochondrial calcium homeostasis and increas-ing the expression levels of cell energy metabolism-re-lated regulatory proteins.This provides an experimen-tal basis for LbGP's potential role in supporting the im-provement of ovarian function.

Objective To explore the mechanism of electroacupuncture in treating rats with urinary incontinence after suprasacral spinal cord injury based on EP3-HCN1 signaling pathway.Methods Totally 48 female SD rats were divided into sham-operation group(12 rats)and modeling group(36 rats,prepared as urinary incontinence model rats after suprasacral spinal cord injury)according to the random number table method.The molded rats were randomly divided into the model group,electroacupuncture group and electroacupuncture+EP3 agonist group after successful modeling,with 11 rats in each group.The electroacupuncture group stimulated"Guanyuan","Zhongji"and"Sanyinjiao"acupoints,and the electroacupuncture+EP3 agonist group injected EP3 agonist sulprostone into the tail vein before electroacupuncture treatment once a day for 7 days.The urodynamic indexes of rats were detected,the morphology of the bladder tissues were observed by HE staining,Western blot and RT-qPCR were used to detect the protein and mRNA expressions of c-Kit,EP3 and hyperpolarized activated cyclic nucleotide gated channel protein(HCN)1 in bladder tissue,respectively.Immunofluorescence was used to detect the positive expressions of c-Kit,EP3 and HCN1 in bladder tissue.Results Compared with the sham-operation group,the model group rats showed an increase in bladder basal pressure and leak point pressure(P<0.01),and a decrease in bladder maximum capacity and compliance(P<0.01);the arrangement of epithelial cells in bladder tissue was disordered,leading to pathological phenomena such as inflammatory cell infiltration,tissue edema and bleeding;the protein and mRNA expressions of c-Kit,EP3 and HCN1 in bladder tissue significantly increased(P<0.05,P<0.01).Compared with the model group,the electroacupuncture group showed a decrease in bladder basal pressure and leak point pressure(P<0.01),while the bladder maximum capacity and compliance increased(P<0.05,P<0.01);the morphology of bladder tissue was relatively normal;the protein and mRNA expressions of c-Kit,EP3 and HCN1 in bladder tissue were all decreased(P<0.05,P<0.01).Compared with the electroacupuncture group,the electroacupuncture+EP3 agonist group showed an increase in bladder basal pressure and leak point pressure(P<0.05,P<0.01),a decrease in bladder maximum capacity and compliance(P<0.05,P<0.01);aggravated pathological damage to bladder tissue;the protein and mRNA expressions of c-Kit,EP3 and HCN1 in bladder tissue increased(P<0.05,P<0.01).Conclusion Electroacupuncture of"Zhongji","Guanyuan"and"Sanyinjiao"alleviates excessive contraction of the bladder detrusor muscle by inhibiting the activation of the EP3-HCN1 pathway,thereby improving urinary incontinence symptoms after suprasacral spinal cord injury in rats.

Objective:To analyze the clinical characteristics of frailty in elderly patients with chronic obstructive pulmonary disease (COPD).Methods:COPD patients aged ≥65 years registered in the RealDTC study from June 2023 to March 2024 were included. Demographic data, history of exacerbations in the past year, exposure to risk factors (smoking, biomass fuel exposure, occupational exposure), modified Medical Research Council (mMRC) dyspnea score, COPD Assessment Test (CAT) score, forced expiratory volume in the first second predicted of percentage (FEV 1%pred), forced expiratory volume in one second (FEV 1) to forced vital capacity (FVC), and comorbidities (bronchial asthma, bronchiectasis, pulmonary tuberculosis, cardiovascular disease, diabetes mellitus) were collected. According to Fried′s frailty phenotype, patients meeting any 3 of the 5 criteria were defined as frail and divided into a frailty group and a non-frailty group. Multivariate regression analysis was used to screen the related factors of frailty in elderly COPD patients, and the receiver operating characteristic (ROC) curve was used to calculate the area under the curve (AUC) of related factors for frailty assessment. Results:A total of 496 elderly COPD patients were included, of which 144(29.0%) had comorbid frailty. The frailty group had lower mass body index (BMI), FEV 1%pred, and FEV 1/FVC, higher mMRC and CAT scores, more exacerbations and hospitalizations in the past year (all P<0.001), and higher proportions of patients with junior high school education or below, Global Initiative for Chronic Obstructive Lung Disease (GOLD) group E, and GOLD grades 3 and 4 (all P<0.05). Multivariate regression analysis showed that low education level ( OR=2.117, 95% CI: 1.119-4.003), low BMI ( OR=0.927, 95% CI: 0.867-0.991), GOLD grade 4 ( OR=4.251, 95% CI: 1.477-12.235), high CAT score ( OR=1.174, 95% CI: 1.127-1.224), and high mMRC score ( OR=4.578, 95% CI: 3.364-6.231) were independent risk factors for frailty in elderly COPD patients (all P<0.05). The ROC curve showed that CAT score (AUC=0.78) and mMRC score (AUC=0.81) had the highest AUC for assessing frailty in elderly COPD patients. Conclusions:Elderly COPD patients with frailty have lower BMI, worse lung function, and more severe symptom burden. The results provide clinical reference for the management of frail elderly COPD patients.

Sepsis is a life-threatening organ dysfunction disease caused by a dysregulated host response to infection.It has com-plex pathophysiological changes,and in severe cases,it can rap-idly develop into septic shock and multiple organ dysfunction or multiple organ failure.At present,the pathological mechanism of sepsis and its induced organ dysfunction is complex and the in-fluencing factors are numerous.So far,there is still a lack of specific and effective treatment strategies.RNA modify-N6-methyladenosine(m6 A)is one of the most common post-tran-scriptional modifications on eukaryotic RNAs.It is involved in the regulation of the occurrence and development of a variety of inflammatory diseases,including sepsis,and even multiple organ dysfunction induced by sepsis by affecting the metabolism of RNAs.It includes cardiac dysfunction,acute lung injury(ALI)and acute kidney injury(AKI).Therefore,this article will dis-cuss the effect of m6A modification on the function of immune cells,and its important role in sepsis and its induced multiple or-gan dysfunction diseases by regulating inflammatory signals,py-roptosis,mitochondrial damage and ferroptosis.This will provide new therapeutic targets and strategies for the clinical prevention and treatment of sepsis and its induced multiple organ dysfunc-tion diseases.