Objective:To study the methylation level of miR-5194 promoter in bladder cancer tissues, and explore the effects of miR-5194 on the proliferation and migration of bladder cancer cells by targeting p21-activated protein kinase 2 (PAK2).Methods:The methylation level of miR-5194 promoter in bladder cancer tissues was analyzed using MethHC database. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-5194 in bladder cancer MGH-U3, EJ, J82, and UMUC3 cells. 5-aza-2′-deoxycytidine (5-Aza-CdR) was used to treat bladder cancer cell lines, and RT-qPCR was used to detect the changes in the expression of miR-5194 in bladder cancer cell lines after 5-Aza-CdR treatment. UMUC3 cells were divided into miR-5194 group and NC group, and miR-5194 or miR-NC were transfected into UMUC3 cells, respectively. Colony formation assay and scratch assay were used to detect the effect of overexpression of miR-5194 on the proliferation and migration of UMUC3 cells. The bioinformatics tool miRGator and dual-luciferase reporter gene experiments verified the targeting relationship between miR-5194 and PAK2. The effect of overexpression of miR-5194 on the expression of PAK2 mRNA in UMUC3 cells was detected by RT-qPCR. The effect of overexpression of miR-5194 on the expression of PAK2 protein, proliferation-related proteins (CDK1, Cyclin B) and migration-related proteins (FOXC2, E47) in UMUC3 cells was detected by Western blotting. The measurement data were expressed as mean ± standard deviation ( ± s), the independent sample t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison among multiple groups. Results:The methylation level of miR-5194 promoter in bladder cancer tissues was significantly higher than that in adjacent tissues ( P<0.01). Compared with the immortalized bladder epithelial cells SV-HUC-1, the expression of miR-5194 in bladder cancer cells was significantly down-regulated ( P<0.01). After 5-Aza-CdR treatment, the expression of miR-5194 in bladder cancer cells was significantly increased ( P<0.01). The number of colonies in miR-5194 group and NC group were 31.30 ± 8.09 and 99.98 ± 10.53, respectively, and the proliferation ability of UMUC3 cells in miR-5194 group was weakened ( P<0.01). The migration rates of UMUC3 cells in miR-5194 group and NC group were (31.50 ± 7.17)% and (76.06 ± 4.86)%, respectively, and the migration ability of UMUC3 cells in miR-5194 group was weakened ( P<0.01). miR-5194 can target bind PAK2 gene ( P<0.01). The relative expression of PAK2 mRNA in UMUC3 cells of miR-5194 group and NC group were 1.02 ± 0.34 and 5.43 ± 0.76, respectively, and miR-5194 could negatively regulate the expression of PAK2 mRNA ( P<0.01). Compared with the NC group, the expression of PAK2 protein, the expression of proliferation-related proteins CDK1 and Cyclin B, and the expression of migration-related proteins FOXC2 and E47 were down-regulated in UMUC3 cells with miR-5194 overexpression. Conclusion:The methylation level of miR-5194 promoter in bladder cancer tissue was significantly increased, and miR-5194 inhibited the proliferation and migration of bladder cancer cells by targeting down-regulation of PAK2 expression in bladder cancer UMUC3 cells.

Objective To construct a predictive model for cerebral small vessel disease (CSVD) MRI burden based on β2-microglobulin (β2-MG) and lipoprotein(a) [Lp(a)], analyze its predictive value, and validate the model. Methods A total of 138 CSVD patients admitted to Anhui No.2 Provincial People’s Hospital from February 2023 to August 2024 were enrolled. Patients were divided into a low-burden group (n=63) and a moderate/severe burden group (n=75) according to the CSVD MRI burden scoring criteria. The related clinical data were compared between the two groups. Binary logistic regression analysis was used to identify independent factors for CSVD moderate/severe MRI burden. A nomogram predictive model was constructed based on these factors and its performance was evaluated. Results The proportions of male patients, as well as those with a history of diabetes or hypertension, were significantly higher in the moderate/severe burden group than those in the low burden group. Additionally, the age of patients in the moderate/severe burden group was significantly older, and the levels of β2-MG, Lp(a), and homocysteine (Hcy) were higher than those in the low burden group (P＜0.01). Binary logistic regression analysis revealed that hypertension, diabetes, β2-MG, and Lp(a) were independent factors for CSVD moderate/severe MRI burden (P＜0.05). The nomogram predictive model based on these four factors had a cut-off value of 0.467 0, with an area under curve (AUC) of 0.838 7 (95%CI 0.760 8-0.916 6) in the training set (n＝97) and 0.854 1 (95%CI 0.742 1-0.966 1) in the internal validation set (n＝41) . The calibration curve demonstrated good agreement between predicted and observed values. Decision curve analysis (DCA) indicated that the nomogram model had good clinical utility. Conclusions The nomogram model based on β2-MG and Lp(a) has high predictive performance in assessing the risk of CSVD moderate/severe MRI burden, with good discrimination and calibration.

Objective:To evaluate the Caprini risk assessment model (Caprini RAM) for predicting lower extremity deep vein thrombosis (DVT) after a spinal cord injury (SCI).Methods:Five hundred and one SCI patients were divided into low- (0-1 points, n=130), medium- (2 points, n=43), and high-risk (≥3 points, n=328) groups according to their Caprini scores. The data covering all 501 included gender, age, cause of injury, injury site, American Spinal Injury Association classification, smoking history, surgical history, concurrent pulmonary or urinary tract infections, indwelling catheterization, comorbid type 2 diabetes or hypertension, D-dimer level, fibrinogen level, prothrombin time (PT), activated partial thromboplastin time, international normalized ratio), red blood cell (RBC) count, white blood cell count, platelet (PLT) count and neutrophil percentage. DVTs were detected using lower extremity ultrasound. The Caprini scores were treated as both categorical and continuous variables alongside the other risk factors in multivariate logistic regressions predicting DVT incidence. Generalized additive models were used for curve fitting and threshold saturation analysis, and log-likelihood ratio tests were applied to evaluate the linear relationships observed between Caprini scores and DVT incidence. Results:When the Caprini score was used as a categorical variable, a high risk score ( OR=7.264), age ( OR=1.050), D-dimer ( OR=1.11) , PT ( OR=1.494), PLT count ( OR=1.004) and lung infection ( OR=1.83) were found to be significant independent predictors of DVT. RBC was a protective factor ( OR=0.509). When the Caprini score was used as a continuous variable, lung infection lost its predictive utility. After adjusting for the risk and protective factors, the Caprini scores and the incidence of DVT showed a significant curvilinear increasing association, with a breakpoint of 3. When the Caprini score was less than 3, the incidence of DVT increased 1.83 times with a 1 point increase in the Caprini score. Beyond 3 the multiplier was 1.06. Conclusions:With the independent risk factors accounted for, Caprini scores demonstrate a curvilinear increasing relationship with DVT risk. Clinically, special attention should be given to SCI patients with Caprini scores ≥3.

Objective:To evaluate the Caprini risk assessment model (Caprini RAM) for predicting lower extremity deep vein thrombosis (DVT) after a spinal cord injury (SCI).Methods:Five hundred and one SCI patients were divided into low- (0-1 points, n=130), medium- (2 points, n=43), and high-risk (≥3 points, n=328) groups according to their Caprini scores. The data covering all 501 included gender, age, cause of injury, injury site, American Spinal Injury Association classification, smoking history, surgical history, concurrent pulmonary or urinary tract infections, indwelling catheterization, comorbid type 2 diabetes or hypertension, D-dimer level, fibrinogen level, prothrombin time (PT), activated partial thromboplastin time, international normalized ratio), red blood cell (RBC) count, white blood cell count, platelet (PLT) count and neutrophil percentage. DVTs were detected using lower extremity ultrasound. The Caprini scores were treated as both categorical and continuous variables alongside the other risk factors in multivariate logistic regressions predicting DVT incidence. Generalized additive models were used for curve fitting and threshold saturation analysis, and log-likelihood ratio tests were applied to evaluate the linear relationships observed between Caprini scores and DVT incidence. Results:When the Caprini score was used as a categorical variable, a high risk score ( OR=7.264), age ( OR=1.050), D-dimer ( OR=1.11) , PT ( OR=1.494), PLT count ( OR=1.004) and lung infection ( OR=1.83) were found to be significant independent predictors of DVT. RBC was a protective factor ( OR=0.509). When the Caprini score was used as a continuous variable, lung infection lost its predictive utility. After adjusting for the risk and protective factors, the Caprini scores and the incidence of DVT showed a significant curvilinear increasing association, with a breakpoint of 3. When the Caprini score was less than 3, the incidence of DVT increased 1.83 times with a 1 point increase in the Caprini score. Beyond 3 the multiplier was 1.06. Conclusions:With the independent risk factors accounted for, Caprini scores demonstrate a curvilinear increasing relationship with DVT risk. Clinically, special attention should be given to SCI patients with Caprini scores ≥3.

Objective:To investigate the effect of interleukin-12(IL-12)on liver lesions and oxidative stress pathway in Sjo-gren's syndrome mice,and to clarify the possible mechanism of liver lesions in Sjogren's syndrome.Methods:Saliva flow rate,liver function,liver pathology and IL-12 level were measured in NOD,IL-12 knockout(IL-12KO)NOD and C57BL/6(B6)mice.Different concentrations of IL-12 and JAK2,TYK2 inhibitors were applied to mouse hepatoma cells.The oxidative stress index of mouse serum and cell culture supernatants of each group were determined.Results:Compared with the other two groups of mice,the NOD mice had significantly higher GOT and GPT(P<0.05),and decreased serum GSH-PX,SOD and CAT(P<0.05).CAT,GSH,GSH-PX and SOD were decreased,while MDA was increased in mice treated with different concentrations of IL-12.JAK2/TYK2 inhibitors reversed regulatory effects of IL-12 on SOD,GSH and MDA in hepatoma cells(P<0.05).Conclusion:IL-12 may aggravated liver damage of Sj?gren's syndrome through promoting oxidative stress of hepatocytes.

Objective:To establish a mortality risk prediction model of severe bacterial infection in children and compare it with the pediatric early warning score (PEWS), pediatric critical illness score (PCIS) and pediatric risk of mortality score Ⅲ (PRISM Ⅲ).Methods:A total of 178 critically ill children were selected from the PICU of the Children's Hospital of Nanjing Medical University from May 2017 to June 2022. After obtaining the informed consent of the parents/guardians, basic information such as sex, age, height and weight, as well as indicators such as heart rate, systolic blood pressure and respiratory rate were collected from all children. A standard questionnaire was used to score the child 24 h after admission to the PICU. The children were divided into the survival and death groups according to their survival status at 28 d after admission. A mortality risk prediction model was constructed and nomogram was drawn. The value of the mortality risk prediction model, PEWS, PCIS and PRISM in predicting the risk of death was assessed and compared using the receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC).Results:Among the 178 critically ill children, 11 cases were excluded due to severe data deficiencies and hospitalization not exceeding 24 h. A total of 167 children were included in the analysis, including 134 in the survival group and 33 in the death group. A mortality risk prediction model for children with severe bacterial infection was constructed using pupillary changes, state of consciousness, skin color, mechanical ventilation, total cholesterol and prothrombin time. ROC curve analysis showed that the AUCs of mortality risk prediction model was 0.888 ( P<0.05). The AUCs of PEWS, PCIS and PRISM Ⅲ in predicting death in children with severe bacterial infection were 0.769 ( P< 0.05), 0.575 ( P< 0.05) and 0.759 ( P< 0.05), respectively. Hosmer-Lemeshow goodness-of-fit test showed the best agreement between risk of death and PEWS predicted morbidity and mortality and actual morbidity and mortality (χ 2 = 5.180, P = 0.738; χ 2 = 4.939, P = 0.764), and the PCIS and PRISM Ⅲ predicted mortality rates fitted reasonably well with actual mortality rates (χ 2= 9.110, P= 0333; χ 2 = 8.943, P= 0.347). Conclusions:The mortality risk prediction model for predicting the death risk has better prognostic value than PEWS, PCIS and PRISM Ⅲ for children with severe bacterial infection.

Objective:To evaluate the efficacy and safety of apatinib in combination with chemoradiotherapy for head and neck squamous cell carcinoma (HNSCC).Methods:37 patients orally received apatinib at 250 mg/d during concurrent chemoradiotherapy until completion of radiotherapy, complete remission assessed by imaging examination, the onset of unacceptable toxicity or death. Baseline characteristics, objective response rates (ORR) and adverse events were assessed in all enrolled patients with complete baseline and safety data. Progression-free survival (PFS) and overall survival (OS) were calculated by Kaplan-Meier method. Prognostic factors were statistically identified using Cox regression models.Results:The ORR was 85%(95% CI: 72%-98%). The median PFS was 17.9 months and the 2-year OS rate was 62%(95% CI: 48%-80%). Ineffective short-term efficacy ( HR=0.035, 995% CI: 0.02-0.652, P=0.025) was an independent risk factor for poor OS. In addition, ineffective short-term efficacy ( HR=0.104, 95% CI: 0.017-0.633, P=0.014) and lymphocytopenia ( HR=17.539, 95% CI: 2.040-150.779, P=0.009) were independent risk factors for poor PFS. Common adverse events (>60%) included lymphocytopenia (76%), leukopenia (68%) and irradiation-induced mucosal injury (65%). The most common treatment-associated grade 3 adverse event was lymphopenia (49%). Conclusions:Apatinib combined with chemoradiotherapy yield significant anti-tumor activity for HNSCC with controllable toxicity. For patients with advanced HNSCC, short-term efficacy and lymphocytopenia may be potential predictors for clinical efficacy of apatinib combined with chemoradiotherapy.

Lung cancer is currently one of the most common malignant tumors in the world. The occurrence and development of lung cancer, especially non-small cell lung cancer (NSCLC), are closely related to the abnormal expression of long non-coding RNA (lncRNA). lncRNA with a transcript of more than 200 nucleotides is involved in chromatin modification, transcription activation, transcription interference and other regulatory processes, and has varying degrees of regulation on the proliferation, migration, and invasion of tumor cells. It is characterized by up-regulation or down-regulation of expression. At present, there are a large number of studies on lncRNA, because lncRNA has considerable application prospects in the diagnosis, clinical treatment, drug resistance research and prognosis evaluation of NSCLC. In this paper, the overview of lncRNA, the up-regulation or down-regulation of NSCLC-related lncRNA expression, NSCLC clinical treatment and drug-resistant lncRNA were summarized.

Objective::Observational studies indicate that insomnia may increase the risk of developing and/or dying from cardiovascular diseases, especially coronary artery disease (CAD). Our purpose is to explore the underlying causal relationship between genetic variants susceptible to insomnia and the risk of CAD by Mendelian randomization analysis.Methods::The study was conducted using publicly available statistical data on genetic variants identified from a genome-wide association meta-analysis of insomnia ( n = 113,006 individuals) and a genome-wide association meta-analysis of CAD ( n = 184,305 individuals), which consisted of both cases and non-cases. The genetic association between variants and CAD was assessed by the variants’ association with insomnia, and estimations were integrated by an inverse-variance weighted meta-analysis. Results::Among the Mendelian randomized analytical sample, 8 variants were associated with insomnia complaints and CAD. And there was no pleiotropic association with the latent confounders. In addition, in the inverse-variance weighted meta-analysis (the estimations combined from the 8 variants), the odds ratio was 1.15 (95% CI: 1.05-1.25; P= 0.002) for CAD, and in the weighted method analysis, the odds ratio was 1.14 (95% CI: 1.03-1.27; P= 0.015) for CAD. Conclusions::All of the data indicated that some valuable variants might involve in the development of CAD by leading the insomnia. Therefore, insomnia might be a causal factor for CAD, and improving the quality of sleep might be a new way for populations with insomnia to prevent CAD.

Objective::Observational studies indicate that insomnia may increase the risk of developing and/or dying from cardiovascular diseases, especially coronary artery disease (CAD). Our purpose is to explore the underlying causal relationship between genetic variants susceptible to insomnia and the risk of CAD by Mendelian randomization analysis.Methods::The study was conducted using publicly available statistical data on genetic variants identified from a genome-wide association meta-analysis of insomnia ( n = 113,006 individuals) and a genome-wide association meta-analysis of CAD ( n = 184,305 individuals), which consisted of both cases and non-cases. The genetic association between variants and CAD was assessed by the variants’ association with insomnia, and estimations were integrated by an inverse-variance weighted meta-analysis. Results::Among the Mendelian randomized analytical sample, 8 variants were associated with insomnia complaints and CAD. And there was no pleiotropic association with the latent confounders. In addition, in the inverse-variance weighted meta-analysis (the estimations combined from the 8 variants), the odds ratio was 1.15 (95% CI: 1.05-1.25; P= 0.002) for CAD, and in the weighted method analysis, the odds ratio was 1.14 (95% CI: 1.03-1.27; P= 0.015) for CAD. Conclusions::All of the data indicated that some valuable variants might involve in the development of CAD by leading the insomnia. Therefore, insomnia might be a causal factor for CAD, and improving the quality of sleep might be a new way for populations with insomnia to prevent CAD.