This study aims to establish an antibody detection method for porcine deltacoronavirus (PDCoV). The recombinant proteins PDCoV-N1 and PDCoV-N2 were expressed via the prokaryotic plasmid pColdII harboring the N gene sequence of the PDCoV strain CH/XJYN/2016. The reactivity and specificity of PDCoV-N1 and PDCoV-N2 with anti-PEDV sera were analyzed after the recombinant proteins were analyzed by SDS-PAGE and purified by the Ni-NTA Superflow Cartridge. Meanwhile, Western blotting and indirect immunofluorescence assay were carried out separately to validate the recombinant proteins PDCoV-N1 and PDCoV-N2. Finally, we established an indirect ELISA method based on the recombinant protein PDCoV-N2 after optimizing the conditions and tested the sensitivity, specificity, and reproducibility of the method. Then, the established method was employed to examine 102 clinical serum samples. The recombinant protein PDCoV-N2 showed low cross-reactivity with anti-PEDV sera. The optimal conditions of the indirect ELISA method based on PDCoV-N2 were as follows: the antigen coating concentration of 1.25 μg/mL and coating at 37 ℃ for 1 h; blocking by BSA overnight at 4 ℃; serum sample dilution at 1:50 and incubation at 37 ℃ for 1 h; secondary antibody dilution at 1:80 000 and incubation at 37 ℃ for 1 h; color development with TMB chromogenic solution at 37 ℃ for 10 min. The S/P value ≥ 0.45, ≤0.38, and between 0.45 and 0.38 indicated that the test sample was positive, negative, and suspicious, respectively. The testing results of the antisera against porcine epidemic diarrhea virus (PEDV), porcine circovirus 2 (PCV2), transmissible gastroenteritis virus (TGEV), foot-and-mouth disease virus (FMDV), and African swine fever virus (ASFV) showed that the S/P values were all less than 0.38. The testing results of the 800-fold diluted anti-PDCoV sera were still positive. The results of the inter- and intra-batch tests showed that the coefficients of variation of this method were less than 10%. Clinical serum sample test results showed the coincidence rate between this method and neutralization test was 94.12%. In this study, an ELISA method for the detection of anti-PDCoV antibodies was successfully established based on the truncated N protein of PDCoV. This method is sensitive, specific, stable, and reproducible, serving as a new method for the clinical diagnosis of PDCoV.
Animals ; Enzyme-Linked Immunosorbent Assay/methods* ; Swine ; Antibodies, Viral/blood* ; Recombinant Proteins/genetics* ; Deltacoronavirus/isolation & purification* ; Coronavirus Infections/virology* ; Swine Diseases/diagnosis* ; Coronavirus Nucleocapsid Proteins ; Sensitivity and Specificity

Ankylosing spondylitis （AS） is an inflammatory autoimmune disease with chronic low back pain as the main clinical manifestation， which mainly affects the axial joints， peripheral joints and various organs. In severe cases， the spine is stiff or deformed， which affects the quality of life and health of patients. The pathogenic factors of AS are complex， which are related to heredity， immunity and intestinal flora. The pathogenesis of AS is not clear yet. Among them， inflammatory reaction， bone destruction and heterotopic ossification are the main pathological features of AS， which play an important role in the disease process of AS. Traditional Chinese medicine has multi-target， multi-channel and multi-component pharmacological effects， which can prevent and treat AS by anti-inflammation， inhibiting bone destruction and preventing heterotopic ossification， and the clinical effect is remarkable， but there is no relevant literature report. Therefore， this review expounds the relationship between inflammatory reaction， bone destruction and heterotopic ossification and the occurrence and development of AS， and summarizes the latest research reports of traditional Chinese medicine in treating AS from anti-inflammatory， inhibiting bone destruction and preventing heterotopic ossification， aiming at providing reference and new ideas and directions for further research on the prevention and treatment of AS by traditional Chinese medicine.

Objective To screen Cuproptosis genes and characteristic genes for differential prognosis in acute mye-loid leukemia(AML)and explore their prognosis in AML as well as their biological roles and correlations in the immune and tumor microenvironment.Methods AML clinical,transcriptome,genomic,and copy number data were downloaded from three major databases,TCGA,GEO,and UCSC,and Cuproptosis genes were collected from published studies.From the perspective of multiomics,the effects of Cuproptosis gene and characteristic gene on survival,immunity,tumor microenvironment,stem cell correlation and drug sensitivity were studied by various bioinformatics methods,meta-analysis and secondary typing.Results One Cuproptosis gene was identified as a differential prognostic gene in AML and five characteristic genes were identified as influencing the prognosis of AML patients by influencing Cuproptosis,and a prognostic model was established.The differential genes were mainly concentrated in mitochondrial activity,REDOX enzyme and energy metabolism.In terms of immunity,macrophage M0,neutrophils,activated memory CD4 T cells and Tregs were positively correlated with risk score,while macro-phage M2,resting mast cells,immature CD4 T cells,helper follicular T cells and memory B cells were negatively correlated with risk score.In terms of tumor microenvironment,the immune cell score of the low-risk group was lower than that of the high-risk group,and in the total score,the tumor microenvironment score of the low-risk group was also lower than that of the high-risk group,indicating that the tumor purity of the high-risk group was lower than that of the low-risk group.However,there was no significant association between stem cells in the high-risk and low-risk groups,and a total of 14 drugs were found to be sensitive to treat AML.Conclusion Cuproptosis gene and characteristic gene are closely related to immune and tumor microenvironment in AML by constructing a prognostic model of AML.

Background To date,pharmacologic therapy is considered the standard first-line treatment for insomnia disorder,but there are still some concerns over the adverse reactions.Repetitive transcranial magnetic stimulation(rTMS)and cognitive behavioral therapy for insomnia(CBT-I)as an alternative to pharmacologic therapy have the advantages of fewer side effects and better patient tolerance in the treatment of chronic insomnia disorder.Objective To explore the clinical efficacy of rTMS and CBT-I on chronic insomnia disorder,so as to provide a novel therapeutic option for the treatment of chronic insomnia disorder.Methods A total of 50 patients with chronic insomnia disorder attending the outpatient clinic of Inner Mongolia Autonomous Region Mental Health Center or community hospital from September 21,2020 to December 16,2021 and fulfilling the International Classification of Sleep Disorders,third edition(ICSD-3)diagnostic criteria were enrolled.Additionally,16 age-and sex-matched healthy controls recruited from the community were set as control group.Patients were randomly divided into rTMS group and CBT-I group,25 cases in each group,and received rTMS or CBT-I intervention for 6 weeks respectively.At enrollment and completion of intervention,patients were subjected to Polysomnography(PSG),Pittsburgh Sleep Quality Index(PSQI),Insomnia Severity Index(ISI)and Repeatable Battery for the Assessment of Neuropsychological Status(RBANS).All participants underwent resting-state functional magnetic resonance imaging(rs-fMRI)scans,and amplitude of low-frequency fluctuation(ALFF)was calculated.The brain regions with statistically different ALFF values between patient group and control group were chosen as regions of interest(ROIs),and whole-brain seed-based functional connectivity analyses were conducted.Results After a 6-week intervention in the two groups,the main effect of time was significant for PSQI(F=41.160,P<0.05),ISI(F=69.615,P<0.05)and RBANS immediate memory(F=47.923,P<0.05),language(F=12.090,P<0.05)and delayed memory indices(F=28.193,P<0.05).A significant main effect of time for total sleep time(F=8.995,P<0.05),a significant main effect of time for sleep efficiency(F=12.414,P<0.05),a significant main effect of group for sleep efficiency(F=4.342,P<0.05)and a significant main effect of time for N1%(F=7.806,P<0.05)were observed.Sleep efficacy in CBT-I group improved significantly from pre-to post-test(t=-2.785,P<0.05).Patients in rTMS group showed increased functional connectivity between the orbital superior frontal gyrus and other regions including left lentiform nucleus putamen(t=4.991,P<0.05),right median cingulate and paracingulate gyri(t=4.471,P<0.05)and right postcentral gyrus(t=4.922,P<0.05),and increased functional connectivity between the orbital superior frontal gyrus and left middle frontal gyrus was found in CBT-I group(t=6.586,P<0.05).Conclusion rTMS and CBT-I may help alleviate insomnia and improve cognitive function of patients with chronic insomnia disorder.

Background The mental health problems of employees in administrative organs and public institutions are highly valued.However,there is a lack of understanding regarding the mental health status of employees in provincial and municipal administrative organs and public institutions in Qinghai region.Objective To analyze the anxiety and depression of employees in provincial and municipal administrative organs and public institutions in Qinghai region,so as to provide references for promoting their mental health and intervening in psychological problems.Methods From October 23 to 28,2022,a total of 3 096 employees in provincial and municipal administrative organs and public institutions in Qinghai region were enrolled using stratified cluster sampling technique,and were assessed using Self-rating Anxiety Scale(SAS)and Self-rating Depression Scale(SDS).Results A total of 1797(58.04％)employees in municipal and above administrative organs and public institutions in Qinghai region completed a valid questionnaire survey.Depressive symptoms were noted in 579(32.22％).SDS scores yielded statistical difference among the selected employees of different ethnicities,types of job positions and educational levels(F=9.074,101.488,4.477,P<0.01),and the severity of depressive symptoms also demonstrated statistical difference among the selected employees with different ethnicities,educational levels and types of job positions(H=49.196,17.028,160.848,P<0.01).Anxiety symptoms were detected in 711(39.57％)employees.SAS scores exhibited statistical difference among the employees with different genders,ages,ethnicities,educational levels,marital status and types of job positions(t=-4.571,F=6.648,3.950,5.243,12.008,22.253,P<0.01).Statistical differences were also illustrated in the severity of anxiety symptoms among the selected employees with different ages,genders,ethnicities,educational levels,marital status and types of job positions(H=24.361,-3.788,28.365,24.268,25.976,56.204,P<0.01).Conclusion About one-third of employees in municipal and above administrative organs and public institutions in Qinghai region may have symptoms of depression and anxiety.The depression symptoms are more severe among Tibetan people,those with low education levels and ordinary civil servants.The anxiety symptoms are more severe among employees aged 30 and below,those with low education levels,unmarried and those in technical positions.

Objective:To describe the current situation of mental health service utilization of community pa-tients with five mental disorders in Inner Mongolia Autonomous Region and provide reference for health education and formulating relevant policies.Methods:The multistage stratified sampling method with unequal probability was used to select a total of 12 315 community residents aged 18 and over in Inner Mongolia Autonomous Region.Using Composite International Diagnostic Interview,mood disorders,anxiety disorders,substance use disorders,intermit-tent explosive disorders,and eating disorders,and health service utilization were investigated.Descriptive statistics was completed by single factor analysis method.Results:The lifetime rates of consultation and treatment of any mental disorder were 18.7％and 10.2％,respectively.The highest proportion of patients received treatment by non-medical professionals was 31.4％,followed by psychiatrists in psychiatric hospital or psychologists in general hospitals.Among the patients,1.1％of them received medication,and 2.5％received psychotherapy.Conclusion:The utilization rate of mental health services in community patients with five mental disorders is relatively low.It is necessary to conduct health education for medical help seeking properly.

Objective:To explore the effects of internet-based group cognitive-behavioral therapy(IGCBT)on the depressive and anxiety symptoms in patients with major depressive disorder(MDD).Methods:Eighty pa-tients with MDD were recruited and randomly divided into IGCBT combined drug therapy group and control group with drug therapy.Blind assessments were performed using the Hamilton Depression Scale(HAMD)and Hamilton Anxiety Scale(HAMA)three times at baseline,4 weeks after treatment and 8 weeks after treatment,respectively.Results:The effective rates of HAMD and HAMA at the end of the 4th and 8th week,and the effective rates of HAMA at the end of the 8th week were higher in the combined treatment group than those in the control group(Ps＜0.05).The total scores of HAMD and HAMA in the combined treatment group and the control group were lower at the end of the 4th and 8th week than those at baseline(Ps＜0.001).The total scores of HAMD and HAMA were lower in the combined treatment group than those in the control group at the baseline,4 weeks and 8 weeks after treatment(Ps＜0.01).Conclusion:It suggests that intemet-based group cognitive-behavioral therapy combined with antidepressant medication has a better effect on relieving depressive and anxiety symptoms in patients with ma-jor depressive disorder than drug therapy alone,and especially has a better effect on improving anxiety symptom.

This paper presents a systematic review of extant research investigating the effectiveness of Internet-based group cognitive behavioral therapy （IGCBT） for depression and anxiety symptoms with an emphasis on the treatment modalities， treatment protocols， efficacy and mechanism， together with a summary of research limitations and suggestions for future research. Cognitive behavioral therapy （CBT） is a structured， short-term and present-oriented psychotherapeutic treatment， its Internet- and group-delivered modalities have led to its wider application， and has proven to be clinically effective in alleviating depression and anxiety symptoms caused by different diseases， whereas some drawbacks have been reported. Thus this paper elaborates the clinical application， therapeutic effect and underlying mechanisms of IGCBT intervention for depression and anxiety symptoms， so as to provide references for its future application. ［Funded by the Natural Science Foundation of the Inner Mongolia Autonomous Region （number， 2019MS03038）

Objective : A convolutional neural network (CNN) model was established to automatically analyze flow cytometry (FCM) data to achieve the preliminary diagnosis of acute myeloid leukemia(AML) , and explore the feasibility of applying CNN model to FCM data analysis. Methods : The exploratory study of CNN application was carried out using the bone marrow FCM data obtained by the FlowRepository database and the Clinical Testing Center of Xinjiang Uygur Autonomous Region People ′ s Hospital , and the data had been clinically confirmed whether AML was present. Among them , the public data was divided into training sets , validation sets and test sets according to 6 ∶ 2 ∶ 2 , and local data was used for external test; In order to adapt the FCM data to the CNN model , an FCM data structure based on the image matrix principle was proposed , and after preprocessing the original data , the variables related to the preliminary diagnosis of AML were extracted , including sidescattered light and the expression levels of CD45 , CD13 , CD33 , HLA⁃DR , CD117 , CD34 , and each variable was written into the matrix. Cell sampling and data augmentation methods were used to increase the sample size of the training set , the keras software package was used to build the LeNet⁃5 CNN model in Python , and the training set and the validation set were used for model training and parameter tuning respectively to evaluate the performance of the model on the test set. Results : The accuracy of CNN to identify AML on the two test sets was 0. 931 , 0. 851 , the sensitivity was 0. 667 , 0. 636 , the specificity was 0. 968 , 0. 940 , and the area under the receiver operating characteristic curve was 0. 940 and 0. 917. Conclusion Based on the proposed FCM data structure , the CNN model can realize the preliminary diagnosis of AML , indicating that CNN has certain application value in FCM data analysis.

Objective: To investigate the short-term clinical efficacy of laparoscopic common bile duct exploration(LCBDE) with primary suture or T tube drainage in the management of choledocholithiasis. Methods: The retrospective cohort study was conducted from January 2014 to December 2018 with the clinical data of patients with choledocholithiasis being analyzed. A total of 863 patients were enrolled in this study. There were 431 males and 432 females. The median age was 60 (range 11 to 94). These patients had received LCBDE with primary suture (n=287) and T tube drainage (n=576) in the Department of General Surgery, the First Affiliated Hospital of Nanjing Medical University. Observation indicators: (1)Preoperative blood biochemistry, including blood serum levels of total bilirubin, direct bilirubin, ALT, AST, GGT. (2) Intraoperative conditions, including operation time, blood loss, diameter of common bile duct, number of common bile duct stone.(3)Short-term postoperative conditions, including postoperative hospital stay, postoperative complications. Measurement data with non-normal distribution were described as M (P₂₅, P₇₅), and comparison between groups was done using Mann-Whitney U test. Comparison of count data between groups were analyzed using the chi-square test. Univariate analysis and subsequent multivariable logistic regression were used to investigated the factors affecting the selection of surgical methods. Results: (1) Blood serum levels of total bilirubin and direct bilirubin in the T tube drainage group were 17.0(12.2, 36.0) μmol/L, 7.6(4.9, 19.0) μmol/L, which were significantly higher than those of the primary suture group[15.7(11.8, 29.7) μmol/L, 6.7(4.4, 16.5) μmol/L)](Z=-2.023, -2.468, P<0.05). Preoperative blood serum levels of ALT, AST and GGT in the T tube drainage group were 56.7 (26.6, 128.8) U/L, 38.0(24.3, 75.8) U/L and 179.7(50.8, 394.4) U/L, the primary suture group were[68.2(24.8, 165.3) U/L, 35.5(22.8, 96.9) U/L and 235.2(74.9, 459.1) U/L], with no difference between the two groups (Z=-0.985, -0.437, -1.740, P>0.05). (2)The operation time of the primary suture group was 85(70, 100) min, which was significantly shorter than that of the T tube drainage group[97(75, 120) min](Z=-5.532, P<0.05). The diameter of common bile duct in the primary suture and T tube drainage group were 1.0(0.8, 1.2) cm and 1.0(0.8, 1.2) cm, respectively. Significant difference was observed between the two groups(Z=-2.071, P<0.05). The intraoperative blood loss in the primary suture and T tube drainage group were 20(10, 50) ml and 20(20, 50) ml, with no difference between the two groups (Z=-0.477, P>0.05). 61.32%(176/287) and 67.36%(388/576) of patients in the primary suture group and T tube drainage group were found with multiple stones in the common bile duct, with no difference between the two groups (χ²=3.083, P>0.05). (3)The primary suture group showed shorter postoperative hospital stay compared with the T tube drainage group[4(3, 5) d vs 6(5, 6) d, Z=-12.057, P<0.05]. The primary suture group showed more patients with bile leakage (2.09%) compared with that of the T tube drainage group (0.35%). Multivariable logistic regression showed that the number of common bile duct stone, diameter of common bile duct, time period of surgery, surgery group were significant factors affecting the selection of surgical methods(OR=1.687, 2.423, 0.587, 4.632, 95%CI: 1.152-2.470, 1.519-3.865, 0.511-0.675, 3.698-5.802, P<0.05). Conclusions Although different surgeons showed different opinions with the method of primary suture, laparoscopic common duct exploration with primary suture is safe and reliable in the management of choledocholithiasis with shorter operation time and faster postoperative recovery. T tube drainage is not absolutely necessary in the management of choledocholithiasis. Patients with multiple common bile duct stone or large diameter of common bile duct are likely to receive T tube drainage.