The WRKYs are a group of plant-specific transcription factors that play important roles in defense responses. In this study, we silenced 2 GmWRKY33B homologous genes using a bean pod mosaic virus (BPMV) vector carrying a single fragment from the conserved region of the GmWRKY33B genes. Silencing GmWRKY33B did not result in morphological changes. However, significantly reduced resistances to Pseudomonas syringae pv. glycinea (Psg) and soybean mosaic virus (SMV) were observed in the GmWRKY33B-silenced plants, indicating a positive role of the GmWRKY33B genes in disease resistance. Kinase assay showed that silencing the GmWRKY33B genes significantly reduced the activation of GmMPK6, but not GmMPK3, in response to flg22 treatment. Reverse transcriptase PCR (RT-PCR) analysis of the genes encoding prenyltransferases (PTs), which are the key enzymes in the biosynthesis of glyceollin, showed that the Psg-induced expression of these genes was significantly reduced in the GmWRKY33B-silenced plants compared with the BPMV-0 empty vector plants, which correlated with the presence of the W-boxes in the promoter regions of these genes. Taken together, our results suggest that GmWRKY33Bs are involved in soybean immunity through regulating the activation of the kinase activity of GmMPK6 as well as through regulating the expression of the key genes encoding the biosynthesis of glyceollins.
Glycine max/genetics* ; Disease Resistance/genetics* ; Biological Assay ; Dimethylallyltranstransferase ; Gene Silencing

Objective:The quality of domestic diagnostic accuracy research was evaluated to explore the quality level of domestic diagnostic accuracy research based on Standards for Reporting of Diagnostic Accuracy 2015 (STARD2015).Methods:The Chinese core journals of CNKI and Wanfang Database were searched, and research literature on diagnostic accuracy published from 2017 to 2022 were collected. The main search terms are diagnostic test, sensitivity, specificity, receiver operating characteristic (ROC) curve, etc. The literature was selected according to the inclusion criteria, and the quality of the included literature was evaluated independently by two reviewers, and the conformity rate of the STARD2015 report and the STARD2015 article was calculated. The report quality of the literature was divided into three levels (low, medium and high) according to the conformity rate of the STARD2015 report. The proportion of literature at each level and the proportion of literature at medium and high level in each year were counted. According to quality analysis on the contents of the included articles, articles were divided into standardized reports and unstandardized reports. Intragroup correlation coefficient ( ICC) was used to analyze the consistency of two reviewers. The trend Chi-square test was used to analyze the trend of the proportion of medium and high level literature in each year. One-way analysis of variance was used to compare the coincidence rates evaluated by STARD2015 for each year. Results:A total of 6 771 studies on diagnostic accuracy published from 2017 to 2022 were included. The compliance rate sccording to STARD2015 was 39.56%±4.90%, and the reported compliance rate ranged from 17.65% to 64.71% (the number of reported items ranged from 6 to 22), and 93.53% (6 333/6 771) literatures were in the middle level. Compliance rate of STARD2015 reports varied significantly among different years ( F=25.023, P<0.01), and the compliance rate of 2021 was significantly higher than that of other years ( P<0.01). The proportion of medium and high level literatures according to STARD2015 showed an increasing trend ( χ 2=14.099, P<0.01). The reporting situation of each item varied significantly, and the conformity rate of items raned from 0 to 100%. According to report item, non-standard report rate was 10.34% (569/5 503) for item 6, 4.15% (277/6 677) for item 8, 21.84% (1 447/6 626) for item 20, 66.67% (24/36) for item 22, and 26.03% (877/3 369) for item 26. Conclusions:The overall report quality of published domestic literature on diagnostic accuracy from 2017 to 2022 is at a medium level according to STARD2015, and the reports conformity rate of each item vary significantly, indicating significant knowledge gap on STARD2015 among domestic researchers. The promotion of STARD2015 needs to be strengthened.

Objective To investigate the differential expression of proteins in the serum of diabetic patients quantitative proteomics technologies, and to identify specific protein biomarkers. Methods Serum samples from three diabetic patients and three healthy controls in Shenzhen in 2023 were collected. Immunoprecipitation and nano liquid chromatography-quadrupole-electrostatic orbitrap-linear ion trap mass spectrometry (easy-nanoLC-Q-orbitrap-fusion) were employed to perform fraction and no-fraction-based quantitative analysis of the serum proteome in diabetic patients and healthy controls. The mass spectrometry data were analyzed by MaxQuant1.6.1.0, and the differential proteins identified by the fraction-based method were subjected to gene ontology (GO) functional clustering and kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Results A total of 251 reliable proteins were identified by the no-fraction-based method, among which 17 differentially expressed proteins were selected. In contrast, 494 reliable proteins were identified by the fraction-based method, and 74 differentially expressed proteins were selected. Among them, seven proteins showed consistent changes in both methods. Five proteins were downregulated, including pregnancy zone protein (PZP), carbonic anhydrase 1 (CAH1), mannose-binding protein C (MBL2), fibrinogen alpha chain (FIBA), and haptoglobin (HPT). Two proteins were upregulated, including apolipoprotein(a) (LPA) and complement factor H-related protein 4 (CFH4). GO analysis revealed that the differentially expressed proteins were mainly involved in molecular functions such as glycosaminoglycan binding and heparin binding. KEGG pathway analysis showed that the differentially expressed proteins were mainly associated with complement and coagulation cascades, cell adhesion, PI3K-Akt signaling pathway, and other pathways. Conclusions Fraction-based no-fractional quantitative proteomics analysis provides an effective approach for analyzing differential protein expression in the serum of diabetic patients, and offers a valuable technique for screening specific protein biomarkers of serum samples in large-scale cohorts of disease.

OBJECTIVES: To explore the role of mitochondrial CYB 15024G>A mutation in the development of essential hypertension. METHODS: Mitochondrial genome sequences of hypertensive patients were obtained from previous studies. Clinical and genetic data of a hypertensive patient with mitochondrial CYB 15024G>A mutation and its pedigree were analyzed. Lymphocytes derived from patient and family members were transformed into immortalized lymphoblastoid cell lines, and the levels of adenosine triphosphate (ATP), mitochondrial membrane potential and intracellular reactive oxygen species (ROS) were detected. RESULTS: The penetrance of this essential hypertension family was 42.9%, and the age of onset was 46-68 years old. Mitochondrial genome sequencing results showed that all maternal members carried a highly conserved mitochondrial CYB 15024G>A mutation. This mutation could affect the free energy of mitochondrial CYB for secondary and tertiary structure and protein folding, thereby changing its structural stability and the structure of the electron transfer function area around the mutation site. Compared with the control, the cell line carrying the mitochondrial CYB 15024G>A mutation showed significantly decreased levels of mitochondrial CYB, ATP and mitochondrial membrane potential, and increased levels of ROS (P<0.01). CONCLUSIONS Mitochondrial CYB 15024G>A mutation may affect the structure of respiratory chain subunits and mitochondrial function, leading to cell dysfunction, which suggests that the mutation may play a synergistic role in essential hypertension.
Humans ; Middle Aged ; Aged ; Reactive Oxygen Species ; Essential Hypertension/genetics* ; Adenosine Triphosphate ; Cell Line ; Mutation

Abstract OBJECTIVE To conduct a quality markers(Q-Markers) study on the treatment of cough with Feire Zhike granules based on the “five principles” of Q-Markers, combined with mass spectrometry, network pharmacology and cobweb model. METHODS Identification of the main chemical components in Feire Zhike granules by liquid mass spectrometry. The TCMSP database was searched to collect the active ingredients and corresponding targets of 11 herbs in the prescription of Feire Zhike granules. The GeneCards database was also searched to collect genes encoding cough-related proteins, and a herb-component-target network map was constructed. Assigning effectiveness to active ingredients based on the number of target sites. Check the pharmacopoeia and literature to obtain the measurable components of each herb. Determine the monarchical, ministerial and adjuvant attribution of the original herbs in the compound formula based on the principle of compatibility. The literature and TCMSP database were searched to screen the original herb for specificity components. RESULTS A total of 43 chemical components in the Feire Zhike granules were identified to be “delivered” to the formulation based on UHPLC-Q-TOF-MS/MS. The 124 active compounds were obtained from the TCMSP database and acted on 120 targets, 93 of which were associated with coughing. There were 6 components in the pharmacopoeia that specify quantitative methods. A total of 26 endemic components were found in 11 herbs in the compound formula. In addition, the above components were ranked according to the compatibility rules of monarch and minister and the scoring rules of cobweb model. The top 10 components were wogonin, baicalein, licochalcone A, nobiletin, acacetin, hesperidin, liquiritin, tuberstemonine, β-sitosterol and galangin, respectively. CONCLUSION The above 10 components can be identified as Q-Markers of Feire Zhike granules.

Objective To observe the value of multimodal imaging for diagnosis of cardiac space-occupying lesions.Methods Data of 70 patients with cardiac space-occupying lesions who underwent echocardiography and cardiac CT(CCT)were retrospectively analyzed,among them 35 also underwent cardiac MRI(CMRI).The value of multimodal imaging for diagnosis of cardiac space-occupying lesions were explored according to the results of surgical pathology or clinical diagnosis.Results Among 70 cases,benign tumors were confirmed by surgical pathology in 43 cases,while malignant tumors were confirmed by surgical pathology in 3 cases and clinically diagnosed in 1 case.Meanwhile,non-tumor-occupying lesions were clinically diagnosed in 23 cases,all obviously shrunken after treatments.Among 70 cases,echocardiography correctly diagnosed 57 cases,misdiagnosed 8 cases and unclearly diagnosed 5 cases,with diagnostic accuracy rate of 81.43%(57/70).CCT correctly diagnosed 63 cases,misdiagnosed 4 cases but missed 3 cases,with diagnostic accuracy rate of 90.00%(63/70).CMRI outcomes in all 35 cases were consistent with surgical pathologic results,with diagnostic accuracy rate of 100%(35/35).Conclusion Multimodal imaging might provide objective evidences for diagnosis and treatment of cardiac space-occupying lesions.

Objective:To investigate the proteins interacting with Mycobacterium tuberculosis Rv1705c in human body. Methods:Rv1705c was prokaryotically expressed and inclusion bodies were collected for further lysis and the purification of Rv1705c. ELISA assay was used to detect the secretion of IFN-γ after stimulating macrophages with Rv1705c protein. Purified and biotin-labeled Rv1705c sample was incubated on the HuProt? human proteome microarray to screen the interacting proteins. GenePix Pro 6.0 software was used to extract all features of the data obtained from the scanned images and further analysis was performed based on bioinformatics databases such as GO and KEGG. GST pull-down was performed to verify the interaction of Rv1705c with PSMA3 and RSAD2.Results:The purification results showed that Rv1705c was expressed in endosomes. The secretion of IFN-γ increased significantly after stimulating macrophages with Rv1705c. A total of 29 potential Rv1705c-interacting proteins were screened, and nine of them showed signal-to-noise ratio (SNR)>1.6, namely PSMA3, NLN, THOP1, UPF3A, RSAD2, OMG, PNKD, STEAP3 and MED8. Further bioinformatics analysis revealed that PSMA3, RSAD2 and C1QBP were involved in innate immune signaling pathway, and there were interactions of PSMA3 and RSAD2 with IFN. GST pull-down assay validated that PSMA3 and RSAD2 interacted with Rv1705c.Conclusions:This study showed that PSMA3 and RSAD2 interacted with Rv1705c, providing reference for further investigation on the mechanism of Mycobacterium tuberculosis infection.

ObjectiveTo investigate effect of conducting training of autism spectrum disorder （ASD） early screening skill on improving the ability to early identify ASD of medical staffs in primary care hospitals. MethodsIn September 2021， the training of ASD early screening skills was carried out for medical staffs from 20 primary care hospitals in Chengdu. After training， the training effect was evaluated. The numbers of referrals from primary care hospitals to superior hospitals， confirmed ASD as well as their average diagnostic age of children with ASD before and after training were used as evaluation indicators. ResultsAfter training， the number of children with suspected ASD referred by primary care hospitals was more than that before training ［（16.65±11.60） vs. （3.40±2.23）， t=5.431， P<0.01］， the number of children diagnosed with ASD was more than that before training［（6.85±4.93） vs. （2.45±1.67）， t=4.171， P<0.01］， and the differences were statistically significant. As for the diagnosed age of ASD children， after training， the average age was lower than that before training ［（34.95±11.67） vs. （42.2±14.64）， t=-2.553， P=0.019］. ConclusionTraining of ASD early screening skills for medical staffs in primary care hospitals may help to improve their ability to early screening ASD children.

Objective To retrospectively analyzed the echocardiographic characteristics of ventricular diverticula in fetuses ,children and adults ,and discuss the main points of diagnoses and prognoses of ventricular diverticula . Methods Echocardiographic characteristics ,clinical data and intraoperative findings of ventricular diverticula of 8 fetuses ,3 children and 14 adults were summarized . Results The ventricular diverticula in 8 fetuses and 3 children were all single . Twenty five diverticula were found in the 14 adult patients ;in other words ,there were 3 multiple diverticula found in adult group . A statistical difference was found in ratio of diverticula area and corresponding ventricular area ( P = 0 .021 ) ,and corresponding ventricular fractional shortening ( P =0 .003) between fetal and single adult ventricular diverticula . There were 2 cases with ventricular septal defects ,1 with pericardial effusion ,1 with single umbilical artery ,and 2 with atrioventricular valvular regurgitation in fetal cases . Congenital complex cardiac disease and dextrocardia were found in 1 child . There were 1 case with supravalvular aortic stenosis , 1 with nonobstructive hypertrophic cardiomyopathy ,1 with left ventricular noncompaction ,and 1 with mitral reguigitation in adult group . All the 7 diverticula with ventricular arrhythmias ,ST‐T changes and abnormal Q wave were fibrous type . And 2 multiple diverticula were accompanied with ventricular dysfunction . Among the fetal cases ,2 cases were born ,3 cases were terminated pregnancy ,and 3 cases were lost to follow up . Three children and 2 adults were underwent cardiac operations for diverticula with good outcomes . No such complications as cardiac rupture ,thromboembolism ,and sudden death were found . Conclusions Echocardiography is a useful method to diagnose ventricular diverticulum ,especially for fetuses ,and provide the basis for prognosis consultation .

Objective To investigate the changes and correlation of mitral valve coaptation length index CLI and coaptation area index CAI after mitral valvuloplasty MVP Methods A total of 30 subjects undergoing MVP for mitral regurgitation MR were studied Coaptation length CL CLI coaptation area CA and CAI were determined before and after surgery by 2-dimensional transoesophageal echocardiography 2D-TEE and 3-dimensional transoesophageal echocardiography 3D-TEE Results Compared with preoperative measurements CL CLI CA and CAI were significantly increased in postoperative studies CL 4 7±0 7 mm vs 9 4± 1 1 mm CLI 9 1 ±3 3 vs 38 5 ±4 1 CA 148 9 ± 65 3 mm 2 vs 371 9 ± 144 3 mm 2 CAI 9 3 ±3 1 vs 35 9 ± 7 5 all P < 0 05 CLI was significantly correlated with CAI both preoperatively r = 0 770 P < 0 01 and postoperatively r = 0 771 P <0 01 Furthermore CLI and CAI were significantly negative correlated with the degree of MR r =-0 897 P <0 01 r =-0 886 P <0 01 Conclusions Coaptation variables increased significantly in subjects after MVP CLI by 2D-TEE was related to CAI by 3D-TEE and both were useful for the assessment of mitral valve coaptation But CLI by 2D-TEE was more simple and feasible in clinic.