Background/Aims: Cholangiocarcinoma (CCA) is a primary malignant neoplasm with an extremely poor prognosis. While combined chemoradiotherapy has been demonstrated to delay CCA progression to a certain extent, the absence of specific molecular biomarkers or targets significantly hinders the diagnosis and treatment of CCA. Methods: Through cross-analysis of proteomics and ADMA modificationomics, we identified DDX1 overexpressed in CCA with elevated R602-ADMA modifications. HPLC-MS/MS identified PRMT1 as the methyltransferase and USP10 as the deubiquitinating enzyme for DDX1. Immunofluorescence and nuclear-cytoplasmic partitioning experiments confirmed DDX1’s nuclear localization. GO and KEGG analyses clarify the biological functions of DDX1 in response to hypoxia. RNA-seq transcriptomics analyzed key pathways influenced by DDX1. A hydrodynamic in situ CCA mouse model was established to validate the chemopreventive effects of the PRMT1-specific inhibitor GSK715 on CCA development. Results: DDX1 promotes CCA progression both in vivo and in vitro and can be inhibited by GSK715. Mechanistically, PRMT1 mediates ADMA modification at position R602 of DDX1. This modification promotes DDX1 nuclear localization by recruiting USP10 to deubiquitinate DDX1, while simultaneously inhibiting PRMT1 degradation. DDX1 promotes the transcription of PRMT1 and USP10 by binding to the mRNA 3’UTR region, establishing a positive feedback regulatory pathway. This mechanism promotes the occurrence and development of CCA and can serve as a target for the inhibitor GSK715 to suppress CCA progression. Conclusions Our study identified DDX1-R602-ADMA modification as a novel ADMA modification in CCA. It further confirmed its pivotal role in CCA progression. Targeting the USP10-PRMT1-DDX1 axis may represent a significant therapeutic approach for CCA.

ObjectiveTo develop and validate a nomogram for predicting the risk of concurrent cognitive impairment in older adults with heart disease. MethodsSelf-reported data from 2 673 respondents in the 2020 China Health and Retirement Longitudinal Study were analyzed. The participants were randomly divided into a training set (n = 1 871) and a validation set (n = 802). Heart disease status was determined using a self-report questionnaire. Logistic regression analysis was performed to identify predictors, and a nomogram for predicting the risk of concurrent cognitive impairment in older adults with heart disease was developed using R software. Bootstrap resampling was used for internal validation, and model performance was further evaluated in the randomly split validation set. The predictive performance of the model was assessed using the area under the receiver operating characteristic curve (AUC) and calibration curves.. ResultsA total of 2 673 older adults with heart disease were included, out of whom 315 participants in the training set had cognitive impairment. Lasso regression analysis identified age, education level, place of residence, instrumental activities of daily living and depression as predictors of concurrent cognitive impairment. A prediction model incorporating these five variables was constructed. The AUC were 0.827 (95%CI 0.804 to 0.850) in the training set and 0.847 (95%CI 0.812 to 0.882) in the validation set. The Hosmer-Lemeshow test yielded P = 0.154 and P = 0.716 in the training and validation sets, respectively. Calibration curves showed good agreement between predicted and observed probabilities. Decision curve analysis indicated that the model provided favorable net benefit and predictive performance. ConclusionThe nomogram may provide a useful reference for risk screening and early intervention for cognitive impairment in older adults with heart disease.

Objective To investigate the therapeutic effects and mechanisms of Jianyang Tablets(mainly with the functions of tonifying the kidneys,replenishing essence,invigorating yang,and alleviating erectile dysfunction)on erectile dysfunction(ED)in rats with liver depression and kidney deficiency syndrome.Methods The 60 male SD rats were randomly divided into a normal group,a model group,a Tadalafil group,and low-,medium-,and high-dose Jianyang Tablets groups.Except for the normal group,all other groups underwent a 14-day composite modeling protocol combining intramuscular hydrocortisone injections and limb immobilization to induce ED with liver depression and kidney deficiency syndrome.Treatments were administered for 28 consecutive days after successful modeling.The body mass differences were recorded and compared before and after the experiment.Behavioral assessments included open-field test and mounting test were conducted.Enzyme-linked immunosorbent assay(ELISA)was used to measure nitric oxide synthase(NOS),cyclic adenosine monophosphate(cAMP),and cyclic guanosine monophosphate(cGMP)levels in penile cavernous tissues.Nitric oxide(NO)level was quantified using the Griess reagent colorimetric method.Results Compared with the normal group,the model group exhibited reduced horizontal activity grid counts in the open-field test,prolonged mounting latency,decreased mounting frequency,decreased body mass,and reduced levels of NOS,NO,cGMP,and cAMP in penile cavernous tissues,with statistical significance(P＜0.05 or P＜0.01).Compared with the model group,the Tadalafil group and medium-and high-dose Jianyang Tablets groups showed increased horizontal activity grid counts and vertical activity counts in the open-field test,shortened mounting latency,elevated mounting frequency,increased body mass,and upregulated levels of NOS,NO,cGMP,and cAMP in penile cavernous tissues,with statistical significance(P＜0.05 or P＜0.01).No significant differences were observed among the intervention groups for the aforementioned indices(P＞0.05).Conclusion Jianyang Tablets effectively improves erectile function in rats with liver depression and kidney deficiency syndrome,potentially mediated by modulation of the NO-cGMP signaling pathway.

Objective To investigate the expression and clinical significance of lncRNA insulin growth factor 2 anti-sense IGF2-AS in colorectal cancer;To verify the biological function of IGF2-AS in cells and to screen protein mole-cules regulated by IGF2-AS.Methods Using bioinformatics to predict the expression and clinical significance of IGF2-AS in colorectal cancer;Using RT-qPCR to detect the expression of IGF2-AS mRNA;Using small interfering RNA to knockdown IGF2-AS;Using CCK8,Transwell assay and flow cytometry to detect proliferation,migration,and apoptosis,respectively;Using label free quantitative proteomics to screen for factors regulated by IGF2-AS;Using databases such as GenCards,OMIM,TTD,and proteomics results screening for core targets regulated by IGF2-AS in colorectal cancer.Results Bioinformatics analysis showed that IGF2-AS was significantly upregulated in colorectal cancer(P＜0.001),its expression might be related to TNM stage(P＜0.01)and the presence of lymph node metastasis(P＜0.001);RT-qPCR results showed that as compared to the control group,the expression level of IGF2-AS mRNA was significantly up-regulated in colorectal cancer tissues(P＜0.01)and most colon cancer cells(P＜0.000 1);Clinical pathological data indicated that the expression of IGF2-AS was potentially asso-ciated with the lymph node metastasis(P＜0.05);The proliferation rate and migration rate of colorectal cancer cells transfected with IGF2-AS siRNA were significantly reduced(P＜0.000 1),and the apoptosis rate was signifi-cantly increased(P＜0.05);There were 13 proteins including KRAS,TCF7L2 and CASP3,that were potentially core targets regulated by IGF2-AS.Conclusions IGF2-AS is significantly up-regulated in colorectal cancer and may play a cancer promoting role in colorectal cancer development.

Objective To investigate the association between serum magnesium levels,serum phosphorus concentrations,vascular calcification,and cardiovascular disease mortality in maintenance hemodialysis patients.Methods This study enrolled 200 hemodialysis patients admitted to Nanjing Drum Tower Hospital from May 2020 to May 2022 as subjects,with an additional 200 healthy individuals from the same period selected as a control group.The biochemical indicators between the two groups were compared;their correlations were analyzed.Binary logistic regression was used to investigate the independent factors of serum magnesium and phosphorus levels in relation to vascular calcification and cardiovascular events in maintenance hemodialysis patients.ROC curve analysis was employed to assess the predictive value of serum magnesium and phosphorus for vascular calcification and cardiovascular events.Results The research group's patients exhibited significantly elevated levels of blood phosphorus,calcium-phosphorus product,iPTH,AACS,and 25-(OH)-VitD compared to the control group.In contrast,their blood magnesium and BMP-7 levels were notably lower than those of the control group,with statistical significance(P<0.05).Pearson correlation showed positive correlations between serum magnesium and serum calcium,phosphorus,calcium-phosphorus product,25-(OH)-VitD3,and BMP-7(r=0.385,0.183,0.141,0.131,0.458,P<0.05);between serum calcium and serum phosphorus,calcium-phosphorus product,iPTH,AACS,25-(OH)-VitD3,and BMP-7(r=0.318),correlation(r=0.318,0.311,0.098,0.170,0.277,0.485,P<0.05);between serum phosphorus and calcium-phosphorus product,iPTH,AACS,25-(OH)-VitD3(r=0.362,0.506,0.367,0.461,P<0.05);between calcium-phosphorus product and iPTH,AACS,25-(OH)-VitD3(r=0.542,0.373,0.434,P<0.05);between iPTH and AACS,25-(OH)-VitD3 showing positive correlations(r=0.553,0.616,P<0.05)and a negative correlation with BMP-7(r=-0.373,P<0.05);between AACS and 25-(OH)-VitD3 showing a positive correlation(r=0.402,P<0.05),and a negative correlation with BMP-7(r=-0.155,P<0.05),with statistically significant differences(P<0.05).Statistically significant differences were noted between the two groups in age,diabetes,serum magnesium,serum calcium,serum phosphorus,calcium-phosphorus product,25-(OH)-VitD3,and hs-CRP(P<0.05).Logistic regression analysis showed that age,serum magnesium,serum calcium,serum phosphorus,calcium phosphate product,25-(OH)-vitamin I were all risk factors for cardiovascular disease(CVD)mortality in maintenance hemodialysis(MHD)patients(P<0.05).ROC curve analysis showed that serum magnesium,serum calcium,and serum phosphorus had predictive areas under the curve(AUC)of 0.895,0.802,and 0.851 for CVD mortality in MHD patients,with sensitivities and specificities of 87.5%/98.7%,66.7%/90.8%,and 72.9%/100%,respectively.The combined prediction for CVD mortality in MHD patients showed an AUC of 0.921,with a sensitivity of 81.3%and specificity of 93.4%.Conclusion MHD patients exhibit low blood magnesium levels,elevated serum phosphorus concentrations,and increased calcium-phosphorus product,with complex correlations among these biomarkers.Age,magnesium,calcium,and phosphorus levels were all associated with CVD mortality.ROC curve analysis demonstrates that magnesium,calcium,and phosphorus,both individually and in combination,have high predictive value for CVD mortality risk.

Objective To develop and validate a scoring system to predict the possibility of laparoscopic cholecystectomy (LC) conversion to laparotomy based on preoperative clinical data, and to establish a grading management model of surgery. Methods A retrospective analysis was conducted on the clinical data of 9 414 patients who underwent LC at Renhe Hospital and Huashan Hospital from June 2013 to June 2018. The patients were divided into two groups: the LC group (9 246 patients who successfully underwent LC) and the conversion to laparotomy group (168 patients who required conversion to open surgery). The data of two groups were compared, and the risk factors affecting conversion to laparotomy were screened out by single factor analysis of Chi-square test. Then, the risk factors were analyzed by multiple Logistic regression, and the pre-coefficient of each variable of the risk factors was assigned according to the established conversion to laparotomy possibility function. After calculating the score of each case, the difference in the actual conversion rate of each group was compared. The area under receiver operating characteristic (ROC) curve was calculated to evaluate the performance of the scoring system. According to the scoring system, LC surgical grading management model was created and verified. Results The following factors were identified as significant risk factors for conversion to laparotomy (P < 0.001): body temperature ≥ 38.5℃, frequency of acute cholecystitis ≥3 times, maximum thickness of gallbladder wall ≥ 5 mm, gallbladder neck stone incarceration, diameter of common bile duct ≥8 mm, and surgical experience ≤50 cases were the risk factors for conversion to laparotomy (P < 0.001). A score >3 points was associated with a high risk of conversion to laparotomy. Conclusions The LC scoring system and surgical grading management are reliable and effective tools for predicting and reducing the conversion rate of LC to laparotomy.

ObjectiveTo explore the clinical safety of Feilike Mixture （FLK） in the real world. MethodsThe safety of all children who received FLK from 29 institutions in 12 provinces between January 21，2021 and December 25，2021 was evaluated through prospective centralized surveillance and a nested case control study. ResultsA total of 3 035 juveniles were included. There were 29 research centers involved，which are distributed across 12 provinces，including one traditional Chinese medicine （TCM） hospital and 28 general hospitals. The average age among the juveniles was （4.77±3.56） years old，and the average weight was （21.81±12.97） kg. Among them，119 cases （3.92%） of juveniles had a history of allergies. Acute bronchitis was the main diagnosis for juveniles，with 1 656 cases （54.46%）. FLK was first used in 2 016 cases （66.43%），and 142 juvenile patients had special dosages，accounting for 4.68%. Among them，92 adverse drug reactions （ADRs） occurred，including 73 cases of gastrointestinal system disorders，10 cases of metabolic and nutritional disorders，eight cases of skin and subcutaneous tissue diseases，two cases of vascular and lymphatic disorders，and one case of systemic diseases and various reactions at the administration site. The manifestations of ADRs were mainly diarrhea，stool discoloration，and vomiting，and no serious ADRs occurred. The results of multi-factor analysis indicated that special dosages （the use of FLK）［odds ratio （OR） of 2.642， 95% confidence interval （CI） of 1.105-6.323］，combined administration： spleen aminopeptide （OR of 4.978， 95%CI of 1.200-20.655），and reason for combined administration： anti-infection （OR of 1.814， 95%CI of 1.071-3.075） were the risk factors for ADRs caused by FLK. Conclusion92 ADRs occurred among 3 035 juveniles using FLK. The incidence of ADRs caused by FLK was 3.03%，and the severity was mainly mild or moderate. Generally，the prognosis was favorable after symptomatic treatment such as drug withdrawal or dosage reduction，suggesting that FLK has good clinical safety.

Idiopathic pulmonary fibrosis (IPF), a chronic interstitial lung disease, is characterized by aberrant wound healing, excessive scarring and the formation of myofibroblastic foci. Although the role of alternative splicing (AS) in the pathogenesis of organ fibrosis has garnered increasing attention, its specific contribution to pulmonary fibrosis remains incompletely understood. In this study, we identified an up-regulation of serine/arginine-rich splicing factor 7 (SRSF7) in lung fibroblasts derived from IPF patients and a bleomycin (BLM)-induced mouse model, and further characterized its functional role in both human fetal lung fibroblasts and mice. We demonstrated that enhanced expression of Srsf7 in mice spontaneously induced alveolar collagen accumulation. Mechanistically, we investigated alternative splicing events and revealed that SRSF7 modulates the alternative splicing of pyruvate kinase (PKM), leading to metabolic dysregulation and fibroblast activation. In vivo studies showed that fibroblast-specific knockout of Srsf7 in conditional knockout mice conferred resistance to bleomycin-induced pulmonary fibrosis. Importantly, through drug screening, we identified lomitapide as a novel modulator of SRSF7, which effectively mitigated experimental pulmonary fibrosis. Collectively, our findings elucidate a molecular pathway by which SRSF7 drives fibroblast metabolic dysregulation and propose a potential therapeutic strategy for pulmonary fibrosis.

Objective To investigate whether Dauricine(Dau)can ameliorate acute kidney injury induced by renal ischemia-reperfusion(IR)in mice.Methods C57BL/6 mice were randomly assigned to three experimental groups:sham operation,ischemia-reperfusion injury(IRI),and IRI treated with daunorubicin(IRI+Dau),with 12 animals in each group.Following oral administration of Dau(15 mg/kg),renal ischemia-reperfusion was induced,and blood and kidney tissue samples were collected 24 hours post-surgery.Histopathological changes were assessed using hematoxylin and eosin(HE)staining.Renal function was evaluated by measuring serum creatinine and blood urea nitrogen(BUN)levels.Protein expression related to lipid peroxidation was analyzed using western blotting and immunofluorescence.Inflammatory gene expression was determined via quantitative polymerase chain reaction(qPCR).Nuclear translocation of nuclear factor κB(NF-κB),a key inflammatory marker,was assessed using immu-nofluorescence.Statistical comparisons between groups were performed using t-tests.Results The administration of Dau significantly ameliorated IR-induced acute kidney injury compared to the Sham group.Serum creatinine(P<0.001)and urea nitrogen(P<0.000 1)levels were markedly decreased in Dau-treated mice relative to those in the IRI group.Furthermore,Dau significantly suppressed lipid peroxide production in renal tissues(P<0.001),without significantly affecting the expression levels of Gpx4(P=0.919)and Acsl4(P=0.086),two key proteins involved in lipid peroxidation.In addition,Dau effectively inhibited IR-induced nuclear translocation of NF-κB(P<0.001)and reduced apoptosis in kidney cells(P=0.004).Conclusion Dau mitigates IR-induced kidney damage by reducing the accumulation of lipid peroxides and inhibiting the nuclear translocation of NF-κB,thereby attenuating inflammation and renal cell apoptosis.

AIM:To establish a stable and efficient rat model of alcoholic liver disease(ALD),we investigat-ed the effects of different alcoholic beverage concentrations and alcohol dosing regimens.METHODS:(1)SPF-grade male SD rats were randomized into 5 groups(n=10):blank,ALD1,ALD2,ALD3,and ALD4.Except for the blank group,rats received intragastric administration of 56%alcohol(6 mL/kg twice daily with an 8-hour interval)for 4 weeks,along with free access to 0%,5%,10%,or 15%alcoholic beverage to evaluate concentration-dependent effects.(2)An-other cohort was divided into three groups(n=10):blank,ALD5,and ALD6.Rats(except blank)were gavaged with 56%alcohol twice daily for 9 weeks(8 mL/kg for ALD5;6 mL/kg in week 1,increasing by 0.5 mL/kg weekly for ALD6),with 10%alcoholic beverage available ad libitum to assess dose-dependent effects.Serum biochemical markers[alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglycerides(TG),high-density li-poprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)]and inflammatory cytokines[interleu-kin-6(IL-6),IL-1β and tumor necrosis factor-α(TNF-α)]were analyzed after modeling,complemented by imaging(B-ultrasound,CT,and MRI).Success and survival rates were calculated.RESULTS:(1)ALD1～4 groups exhibited sig-nificantly elevated ALT,AST,TC,TG,LDL-C,IL-1β,IL-6 and TNF-α(P<0.05 or P<0.01)and reduced HDL-C and liver-to-spleen CT density ratio vs blank.ALD3(10%alcoholic beverage)showed the highest modeling success rate with low mortality.(2)ALD5 and ALD6 groups also had siginificant differin terms(P<0.01),with ALD6(gradually increas-ing dose)displaying more severe liver injury,higher success rate,and better survival.CONCLUSION:The optimal ALD model was induced by intragastric administration of 56%alcohol(6 mL/kg twice daily in week 1,increasing by 0.5 mL/kg weekly for 9 weeks)combined with 10%alcoholic beverage.This protocol offers a reliable approach for ALD re-search and drug development.