Objective To investigate the regulatory role of ubiquitin-specific protease 10(USP10)on the protein expression of Krüppel-like factor 4(KLF4)and its impact on the proliferation and invasion ability of hepatocellular carcinoma(HCC)cells.Methods The protein expression differences of USP10 and KLF4 in normal liver cell line L02 and HCC cell lines,including HepG2,HUH7,HCCLM3 were detected by immunoblotting(Western blot)methods.HCCLM3 and HUH7 cells were selected,and lentiviral particles overexpressing or silencing USP10(oe-USP10 or sh-USP10)was transfected into the cells,and they were designated as the oe-USP10 group and oe-NC group,respectively.Immunoprecipitation(Co-IP)experiments were conducted to examine whether USP10 could di-rectly interact with KLF4 in HCCLM3 or HUH7 cells.The Co-IP assay was repeated in HCC cells transfected with oe-USP10 or sh-USP10,with the addition of the proteasome inhibitor MG132,which used to detect the ubiquitina-tion level of KLF4 protein in the transfected HCC cells.The pcDNA3.1 vector containing overexpressed KLF4 or its negative control plasmid(pc-KLF4 or pc-NC)was co-transfected into cells of the sh-USP10 group or sh-NC group.These cells were designated as the sh-NC+pc-NC group,sh-USP10+pc-NC group,sh-NC+pc-KLF4 group,and sh-USP10+pc-KLF4 group.The cell proliferation activity of each group was measured using the CCK-8 assay,and the cell invasion ability was assessed using the Transwell assay.Results Compared to L02 cells,the protein expres-sion of USP10 and KLF4 significantly decreased in HepG2,HUH7,HCCLM3,and other cells(P＜0.05).In HC-CLM3 and HUH7 cells,USP10 protein directly interacted with KLF4.Furthermore,treatment with MG132 resulted in a time-dependent increase in KLF4 protein expression in HCCLM3 and HUH7 cells.Silencing USP10 increased the ubiquitination of KLF4 in HCCLM3 or HUH7 cells,while overexpressing USP10 decreased the ubiquitination level of KLF4 in cells.Compared to the sh-NC+pc-NC group,both the proliferation activity and invasion ability of HCCLM3 and HUH7 cells significantly increased in the sh-USP10+pc-NC group(P＜0.01),while they signifi-cantly decreased in the sh-NC+pc-KLF4 group and sh-USP10+pc-KLF4 group(P＜0.05).Compared to the sh-USP10+pc-NC group,the proliferation activity and invasion ability of cells significantly decreased in the sh-USP10+pc-KLF4 group(P＜0.05).Conclusion USP10 can promote the stability of KLF4 protein through deubiquiti-nation in HCC cell lines,thereby inhibiting the proliferation and invasion of tumor cells.

Objective To investigate the effect of melanoma associated antigen D1 (Mage-D1)on mouse femoral bone mass and mineralization ability of mouse bone marrow mesenchymal cells (BMSCs)and its potential molecular mechanism.Methods Female Mage-D1 gene knockout heterozygous mice and male wild-type (WT)mice were subjected as parent mice to breed Mage-D1 gene knockout homozygous (Mage-D1 KO)mice.PCR and agarose gel electrophoresis were used to identify male Mage-D1 knockout (Mage-D1 KO)mice and littermate male wild-type (WT)mice.Micro-CT scanning was performed to observe mouse femoral bone mass,and ELISA and chemical assay were employed to detect serum levels of calcium,phosphorus,calcitonin,and parathyroid hormone in mice.After primary cultured BMSCs were identified with flow cytometry,immunofluorescence staining was utilized to detect the expression of Mage-D1 in BMSCs.BMSCs were infected by Mage-D1 silencing lentivirus,and then the cells were divided into negative control group (sh-NC)and silencing group (sh-Mage-D1).Cell scratch assay was conducted to detect the migration ability of BMSCs,and flow cytometry and CCK-8 assay were conducted to detect the cycle change and proliferation ability of BMSCs.After mineralization induction,alkaline phosphatase (ALP) staining and alizarin red staining were performed;RT-qPCR and Western blotting were used to measure the expression levels of ALP,Runx2 and Col1.RT-qPCR was used to detect mineralization-related genes p75NTR and Msx1.Results Compared with the WT mice,the femoral cortical bone thickness,cortical bone mineral content,cancellous bone mineral content,trabecular number,and cancellous bone surface density were decreased,and trabecular separation was increased in the Mage-D1 knockout homozygous mice (P<0.05).There were no significant changes in the serum levels of calcium,phosphorus,calcitonin and parathyroid hormone in mice after Mage-D1 knockout.Mage-D1 was expressed in the whole BMSCs and was highly expressed in the nucleus and perinuclear regions.Compared with the sh-NC BMSCs,the sh-Mage-D1 group had decreased proliferation ability (P<0.01),enhanced migration ability (P<0.01),and decreased expression of ALP,Runx2 and Col1 genes (P<0.05)and protein (P<0.01)after mineralization induction,milder ALP and alizarin red stain,and lower expression levels of p75NTR and Msx1.Conclusion Mage-D1 knockout can significantly reduce femur bone mass in mice.It can promote the proliferation and inhibit migration of BMSCs,and positively regulate their mineralization in vitro,and the p75NTR-Dlx1/Msx1 signaling axis may be involved in the regulation of bone metabolism by Mage-D1.

Objective:To gain a deep understanding of the current cognition and training needs of nursing undergraduates regarding their emergency response teamwork skills, and to provide reference for the development of courses on emergency response teamwork among nursing undergraduates.Methods:From September to October 2023, purposive sampling was used to select 15 senior nursing undergraduates from Peking Union Medical College, Beijing University of Chinese Medicine, and Beijing City University as subjects for semi-structured interviews. Colazizzi 7-step analysis method was used to summarize and extract themes.Results:Three themes were extracted, including insufficient cognition and skill in emergency response, lack of emergency response teamwork cultivation, and the need for systematic and comprehensive training courses.Conclusions:Universities, hospitals, and other training institutions should work together to develop a systematic emergency response teamwork training course for nursing undergraduates, to cultivate the skills of nursing undergraduates and reserve talents for high-quality emergency response nursing teams.

Objective:To explore the factors influencing the pulmonary function of persons with mild to moderate adolescent idiopathic scoliosis (AIS).Methods:Forty-four persons with AIS were tested for their pulmonary ventilation functioning. The indicators were the percentage of forced vital capacity (FVC%), the percentage of forced expiratory volume in the first second (FEV1%) and the percentage of maximum chase volume (MVV%). Data including age, gender, course of disease, dancing habits, exercise habits, body mass index (BMI), Cobb angle, rotation angle, diaphragm thickness, and diaphragm excursion (DE) were also collected. Linear regressions were evaluated to analyze the factors best predicting pulmonary functioning. Spearman correlation coefficients were computed to quantify the correlation between respiratory muscle functioning and pulmonary functioning.Results:The univariate analysis showed that dancing and DE independently influence FVC%, FEV1%, and MVV%. DE was the only independent factor significantly predicting pulmonary functioning for mild to moderate AIS patients. DE was also significantly positively correlated with average FVC%, FEV1% and MVV%.Conclusions:Respiratory training aimed at enhancing DE is necessary to improve the pulmonary ventilation of persons with AIS.

Objective:To investigate the prevalence and risk factors of diabetic peripheral artery disease (PAD) in patients with type 2 diabetes mellitus (T2DM) managed in primary health care in China.Methods:A total of 2 528 T2DM patients were selected using a two-stage cluster random sampling method based on the baseline survey of the "China Diabetic Foot Prevention Model Project." The study was conducted in 2015 among T2DM patients in 8 primary healthcare centers in Changshu county and Jiang'an district of Wuhan, China. Data collection methods included a questionnaire, body measurement, and blood glucose detection. The Ankle-Brachial Index (ABI) is the most widely used noninvasive vascular test. A binary logistic regression model was used to analyze the influence factors.Results:The prevalence of PAD was 11.2% among the diabetic patients managed in primary health care in the two cities. The prevalence of PAD under 55 years old, 55- years old, 65- years old, and ≥75 years old were 7.8%, 6.0%, 12.9% and 22.5%, respectively. Multivariate stepwise logistic regression identified influence factors included older age, higher education level, smoking, drinking, postprandial glucose uncontrol, and prior myocardial infarction or angina. Compared to age <55 years, the odds ratio for PAD were 0.74 for 55- years (95% CI: 0.43-1.28), 1.72 for 65- years (95% CI: 1.05-2.81), 3.56 for 75 years and above (95% CI: 2.07-6.11), respectively. Compared to patients with education in primary school and below, the odds ratio was 1.37 (95% CI: 0.97-1.94), 2.48 (95% CI: 1.73-3.55), 1.99 (95% CI: 1.26-3.13) for those with education levels of junior high school, senior high school, and college, respectively. Current smoking ( OR=1.49, 95% CI: 1.02-2.17), current drinking ( OR=0.45, 95% CI: 0.28-0.71), postprandial glucose uncontrol (2 h postprandial plasma glucose >10.0 mmol/L: OR=1.72, 95% CI: 1.22-2.43), and prior myocardial infarction or angina ( OR=2.32, 95% CI: 1.50-3.61) were influencing factors of PAD. Conclusions:Despite the high prevalence of PAD in diabetes managed in primary health care; multiple risk factors are not effectively aware of and under control. It is urgent to promote ABI screening and standardized management for diabetes, especially in primary health care.

Coronavirus disease 2019(COVID-19),which is caused by SARS-CoV-2,varies with regard to symptoms and mortality rates among populations.Humoral immunity plays critical roles in SARS-CoV-2 infection and recovery from COVID-19.However,differences in immune responses and clinical features among COVID-19 patients remain largely unknown.Here,we report a database for COVID-19-specific IgG/IgM immune responses and clinical parameters(named COVID-ONE-hi).COVID-ONE-hi is based on the data that contain the IgG/IgM responses to 24 full-length/truncated proteins corresponding to 20 of 28 known SARS-CoV-2 proteins and 199 spike protein peptides against 2360 serum samples collected from 783 COVID-19 patients.In addition,96 clinical parameters for the 2360 serum samples and basic information for the 783 patients are integrated into the database.Furthermore,COVID-ONE-hi provides a dashboard for defining samples and a one-click analysis pipeline for a single group or paired groups.A set of samples of interest is easily defined by adjusting the scale bars of a variety of parameters.After the"START"button is clicked,one can readily obtain a comprehensive analysis report for further interpretation.COVID-ONE-hi is freely available at www.COVID-ONE.cn.

Objective:To retrospectively analyze the clinical characteristics and drug resistance among COVID-19 patients with bacterial and fungal infections.Methods:Clinical data of COVID-19 patients whose blood, urine, sputum and alveolar lavage fluid samples were positive for bacteria and fungi were collected in Tongji Hospital from February 10 to March 31, 2020. WHONET5.6 software was used to analyze drug susceptibility test results.Results:A total of 95 COVID-19 patients positive for pathogenic bacteria were enrolled and among them, 23 were non-critical patients and 72 were critical patients. The main symptoms in these patients included fever, cough with sputum, fatigue and dyspnea. Male and female critical patients accounted for 63.89% and 36.11%, respectively. Most of the patients with bacterial and fungal infections were critical type, accounting for 23.61%. The mortality rates of non-critical and critical patients were 13.04% and 61.11%, respectively. A total of 179 strains of pathogenic bacteria were isolated. The positive rate of Escherichia coli in non-critical patients was 37.50%, which was higher than that in critical patients. However, the positive rates of Acinetobacter baumannii and Klebsiella pneumoniae in critical patients were both 29.87%, higher than those in non-critical patients. There was no significant difference in the positive rate of gram-positive bacteria or fungi between non-critical and critical patients. It was noteworthy that the positive rate of Candida parapsilosis in blood samples of critical patients was relatively high, reaching 36.40%. Drug susceptibility test results showed that no carbapenem-resistant Escherichia coli stains were detected and 60.87% of Klebsiella pneumoniae strains were resistant to carbapenems. Acinetobacter baumannii strains were 100% resistant to three antimicrobial drugs. Methicillin-resistant Staphylococcus aureus strains accounted for 71.43%, but no vancomycin-resistant gram-positive cocci were found. Conclusions:Critical COVID-19 patients were mostly male and prone to multiple bacterial and fungal infections. The mortality of critical patients was higher than that of non-critical patients. Critical COVID-19 was often complicated by hospital acquired infections caused by bacteria including Acinetobacter baumannii and Klebsiella pneumoniae with high drug resistance.

Objective:To study the value of metastatic lymph node radio (rN) and pathological lymph node stage (pN) in evaluating the prognosis of patients after radical gastric cancer.Methods:The clinicopathological data of 491 patients who underwent radical gastrectomy in Tantai Yantaishan Hospital from Jan. 2013 to Dec. 2017 were retrospectively analyzed. X-tile software was used to group the metastatic lymph node radio by rN. According to the number of lymph node metastasis, pN stage was performed. The correlation between metastatic lymph node radio and other clinicopathological factors was assessed. The metastatic lymph node radio and the pathological lymph node stage in evaluating the prognosis of patients after radical gastric cancer were compared.Results:(1) X-tile analysis showed that the best cut-off values for the metastatic lymph node radio in this study were 0.14 (14%) and 0.63 (63%) . (2) According to the cut-off value, the 491 patients included in the study were divided into rN1 (256 cases) , rN2 (160 cases) , and rN3 (75 cases) three subgroups. The results of the analysis of differences showed that there were significant differencesbetween the groups in terms of tumor diameter, tumor location, surgical resection range, stage, lauren classification, degree of differentiation, pT, pN, vascular cancer emboulus, nerve invasion, and pathological TNM staging groups. (3) Comparison of rN and pN staging in evaluation of the prognosis of patients after radical gastric cancer: ①Kaplan-Meier survival analysis results showed that rN was better than pN. ② Both single factor and multivariate cox analysis showed that rN was an independent risk factor for the prognosis of gastric cancer. In univariate analysis, rN group HR=3.18 (95% CI 2.63-3.84, P<0.001) , pN stage HR=1.88 (95% CI 1.66-2.15, P<0.001) ; rN group HR=2.21 in multivariate analysis (95% CI 1.73-2.82, P<0.001) , pN staging HR=1.31 (95% CI 0.95-1.79, P=0.095) . ③The time-dependent ROC analysis showed that the prognostic ability of rN was better than pN staging before 52 months of postoperative follow-up, and pN staging was more advantageous after 52 months. ④The Lauren classification was used as a stratification factor for stratified analysis. The Kaplan-Meier survival curve indicated that rN was better than pN staging in intestinal, mixed and diffuse gastric cancer, and the AUC curve showed the prediction of rN in patients with mixed and diffuse gastric cancer was better than pN staging, while pN staging performance was slightly better in patients with intestinal gastric cancer. Conclusions:rN is an independent factor affecting the prognosis of patients after radical gastric cancer surgery. When judging the prognosis of patients within 52 months after radical gastric cancer, rN has a better prognostic value than pN. In patients with mixed and diffuse gastric cancer in the Lauren classification, rN shows better prognostic value.

Objective To explore the correlation between pre-transplantation donor kidney biopsy and short-term renal function after transplantation.Methods This study include 240 kidney transplantation of donation after cardiac death (DCD) from July 2016 to April 2018.Banff's score of donor kidney biopsy was employed for estimating kidney status.Results No significant correlation existed between rate of glomerulosclerosis and delayed graft function (DGF) (P =0.815).The rate of glomerulosclerosis was significantly correlated with 1-week estimated glomerular filtration rate (eGFR) and discharge eGFR (P＜0.05).Based upon the glomerulosclerosis rate,the patients were divided into two groups ＜ 20％ (n =220) and ≥20％ (n =20),there was no significant inter-group difference in DGF,1-week eGFR or discharge eGFR (P＞0.05).Arterial fibrosis was significantly positively correlated with DGF and negatively with 1-week eGFR and discharge eGFR (P＜0.05).Statistically significant inter-group differences existed in 1-week eGFR and discharge eGFR that arterial fibrosis scores ＜ 2 (n =19) and ≥2 (n =41) (P＜0.05).Arteriolar hyalinosis score was negatively correlated with 1-week eGFR and discharge eGFR (P＜0.05).Based upon arteriolar hyalinosis scores,they were divided into two groups ＜ 2 (n =193) and ≥2 (n =47).There were significant inter-group differences in DGF,1-week eGFR and discharge eGFR (P＜0.05).Remuzzi scores were negatively correlated with 1-week eGFR and discharge eGFR (P＜0.05).Interstitial fibrosis was significantly positively correlated with DGF (P＜0.05) and negatively with 1-week eGFR and discharge eGFR (P＜0.05).Conclusions Donor kidney glomerulosclerosis rate affects short-term renal function of recipients after transplantation.However,using 20％ as a threshold value is limited in clinical practice.Arterial intimal fibrosis and arteriolar hyalinosis are important factors affecting short-term eGFR.Recipient kidneys with Remuzzi score ＞ 4 had poor renal function after transplantation.Interstitial fibrosis score may be used as a predictor of postoperative DGF and shortterm renal function recovery.It is expected to be discussed more extensively in literature.

Objective To investigate the characteristics of myocardial injury and its underlying mechanism in rats resuscitated from cardiac arrest. Methods Forty-two male Wistar rats were randomly(random number) assigned into the post-resuscitation (PR) 4 h, PR 24 h, PR 48 h, and sham groups. Ventricular fibrillation was induced by transcutaneous electrical epicardium stimulation and untreated for 6 min, followed by cardiopulmonary resuscitation (CPR). Myocardial function, glucose metabolism, myocardial ultrastructure, the status of mitochondrial permeability transition pore (MPTP) and mitochondrial membrane potential (MMP) were evaluated at different time points. Results Myocardial dysfunction was found at 4 h after restoration of spontaneous circulation (ROSC). The ejection fraction and cardiac output were decreased (all P<0.01), the diastole left ventricular posterior wall became thicker (P<0.01), and the end-diastolic volume was reduced (P<0.05). However, cardiac function was recovered almost completely at 48 h after ROSC. The PR 4 h group had a higher SUVmax, a more obvious decreased absorbance, and a lower MMP than the sham group (all P<0.01), but no statistically significant differences were noted between the PR 48 h group and the sham group (P>0.05). At 4 h and 24 h after ROSC, the mitochondria was swollen and the mitochondrial crista was sparse, but the myocardial ultrastructure was complete. Conclusions Post resuscitation myocardial dysfunction occurs after ROSC and the myocardial dysfunction is completely reversible at 48 h after ROSC, which may be related to the reversibility of myocardial injury and the gradual recovery of mitochondrial structure and function.