Objective To explore the potential cuproptosis-related genes(CRGs)in patients with multiple myeloma(MM)and develop a prognostic model for improving prognosis and revealing features of the MM immune microenvironment.Methods ① Transcriptome sequencing data and clinical information were retrieved from the GSE4581 dataset in the Gene Expression Omnibus(GEO)database and the Cancer Genome Atlas-Multiple Myeloma Research Foundation(TCGA-MMRF)database.The 859 patients from the TCGA-MMRF database were assigned into a training set,and the other 414 ones from the GSE4581 dataset into a validation set.LASSO-Cox and multivariate Cox regression analyses were used to construct prognostic models and calculate risk scores.Based on the median risk score,they were categorized into high-and low-risk cohorts.Time-dependent receiver operating characteristic(ROC)and calibration curves were plotted to assess the predictive performance and accuracy of the model.The differences between the high-and low-risk cohorts were explored using Kaplan-Meier survival curve analysis and immune microenvironment correlation analysis.② RT-qPCR and Western blotting were used to verify the expression of prognostic model genes in MM cell lines and normal bone marrow single-nucleated cells,and CCK-8 assay,flow cytometry,and Western blotting were applied to verify the biological function of UBE2D1 in MM cells.Results ① LASSO-Cox and multivariate Cox regression analyses revealed that the model consisted of 4 genes,CDKN2A[HR=1.60(95%CI:1.24～2.05),P=2.5e-4],PDE3B[HR=1.33(95%CI:1.09～1.62),P=4.2e-3],UBE2D1[HR=1.65(95%CI:1.20～2.26),P=2.1e-3]and COA6[HR=1.35(95%CI:1.07～1.71),P=0.01].In the training set,the time-dependent ROC curves predicted that the area under curve(AUC)value of 1-,3-,and 5-year survival rate was 0.63,0.71,and 0.78,respectively,and in the validation set,the AUC value was 0.656,0.657,and 0.797,respectively.Calibration curve analysis showed excellent agreement in predicting 1-,3-,and 5-year prognosis.In the training set,Kaplan-Meier curves showed that patients in the high-risk cohort had a significantly shorter overall survival(OS)than the low-risk cohort[HR=2.18(95%CI:1.58～3.02),P<0.001],and in the validation set,the high-risk cohort still had a shorter OS than the low-risk cohort[HR=2.45(95%CI:1.49～4.05),P<0.001].Immune correlation analysis revealed that the ratios of immune cells,such as plasma cells and CD4+T cells were significantly lower in the high-risk cohort(P<0.05),and the risk scores were positively correlated with the expression of immune checkpoint CTLA-4,tumor-targeted therapeutic sites TNFSF4 and ENTPD1,and microenvironmental chemokines CXCL16,CCL8,and CCL16(P<0.05).② Remarkable differences were observed in the expression of all 4 prognostic model genes between the MM cell lines and normal bone marrow single-nucleated cells(P<0.05),and knockdown of UBE2D1 notably inhibited the proliferation of MM cells(P<0.05).Conclusion Our prognostic models based on CDKN2A,PDE3B,UBE2D1,and COA6 genes can predict the prognosis of MM patients.The risk scores of the genes are significantly correlated with immune infiltration in the tumor microenvironment,which providing new molecular markers for individualized therapy.

Humans ; Asthma/drug therapy* ; Biological Therapy

Objective:To survey the knowledge levels about diagnosis and management of asthma among general practitioners in community health centers in Shanghai and to analyze influencing factors.Methods:A questionnaire survey was conducted from May to June 2022 among general practitioners from 80 community health care centers in Shanghai. The questionnaire contained the basic information of the responders; knowledge about the diagnosis of asthma (symptoms, diagnostic methods, interpretation of bronchodilation and fractional exhaled nitric oxide, FeNO tests); and treatment of asthma (choosing and usage of therapeutic drugs). Logistic regression was used to analyze.Results:A total 324 general practitioners completed the questionnaire survey. Among them 221 (68.21%) had bachelor degree; 200 (61.73%) were attending doctors; the median working duration was 11 years; 174 (53.70%) had training on asthma knowledge in last 3 years. The survey results showed that 55 (16.98%) responders misinterpreted results of bronchodilation test, and 90(5.86%)misinterpreted results of FeNO test for diagnosis of asthma; 244 responders (75.31%) suspected implication of long-term cough for diagnosis of asthma; 277(85.49%)would order bronchodilation text to suspected patients; 273(84.26%)prescribed inhaled corticosteroid plus long-acting beta-2 agonist to asthma patients. For mild asthma patients, 144 responders(44.44%)suggested to use budesonide formoterol as needed; for moderate to severe asthma, 174(53.70%)suggested to use budesonide formoterol regularly and as needed, 100(30.86%)suggested to use salmeterol fluticasone regularly and salbutamol as needed. Logistic analysis showed that longer working duration was correlated with higher misinterpretation rate of bronchodilation and FeNO tests( OR=0.798,95% CI:0.694-0.918, P=0.002; OR=0.859,95% CI:0.739-0.998, P=0.047). The accuracy rate of all the questions was significantly higher in general practitioners who had training on asthma knowledge than those who didn′t receive training in last 3 years(all P<0.05). Conclusions:Most general practitioners in community health service centers in Shanghai have relatively high levels of knowledge about diagnosis and treatment of asthma. Years of work experience and training experience can affect the levels of asthma knowledge among community general practitioners.

Objective To establish a porcine allogeneic left lung orthotopic transplantation model to closely simu-late human lung transplantation.Methods Twelve Huanjiang mini-pigs were used as donors and 12 Bama mini-pigs as recipients.The left lung orthotopic transplantation was completed by the left fourth intercostal thoracotomy.At 1 h, 2 h, 4 h, 6 h, 12 h after transplantation, the left and right pulmonary artery pressure were measured, the left and right pulmonary vein blood gas was analyzed, and samples of the left and right lung tissues were taken to determine the water content and for pathological examination.Results All animals survived, and the transplanted pulmonary vein blood PaO2/FiO2 and PAP were rised along with the prolonged postoperative time, compared with those of the recipient normal lung showing a signifi-cant difference (P<0.05).With the pass of time, there were increasing edema, inflammatory cell infiltration, RBC ooze, thickening of alveolar wall in the transplanted lung tissue, and some alveolar lumen occlusion and lung tissue consolidation. The water content of the transplanted lung tissue was increased significantly compared with that in the recipient lung tissue ( P<0.05 ) .Conclusions The established method in this study provides an ideal animal model for research on lung transplantation ischemia-reperfusion injury and immune rejection mechanism.