Flowering and bolting are important agronomic traits in cruciferous crops such as Brassica juncea. Timely flowering can ensure the crop organ yield and quality, as well as seed propagation. The WRKY family plays an important role in regulating plant bolting and flowering, while the function and mechanism of WRKY12 in B. juncea remain unknown. To explore its function and mechanism in bolting and flowering of B. juncea, we cloned and characterized the BjuWRKY12 gene in B. juncea and found that its expression levels were significantly higher in flowers and inflorescences than in leaves. BjuWRKY12 belonged to the Ⅱc subfamily of the WRKY family, and subcellular localization indicated that the protein was located in the nucleus. Ectopic overexpression of BjuWRKY12 in transgenic lines promoted bolting and flowering, leading to significant increases in the expression levels of flowering integrators SOC1 and FUL. Furthermore, yeast one-hybrid and dual luciferase reporter system assays confirmed that BjuWRKY12 directly bound to the promoters of BjuSOC1 and BjuFUL, undergoing protein-DNA interactions. This discovery gives new insights into the regulation network and molecular mechanisms of BjuWRKY12, laying a theoretical foundation for the breeding of high-yield and high-quality varieties of B. juncea.
Mustard Plant/metabolism* ; Flowers/growth & development* ; Plant Proteins/physiology* ; Promoter Regions, Genetic/genetics* ; Gene Expression Regulation, Plant ; Plants, Genetically Modified/genetics* ; Transcription Factors/metabolism* ; MADS Domain Proteins/metabolism*

To rapidly detect and differentiate Mycoplasma gallisepticum(MG)and Mycoplasma synovialis(MS),two sets of specific primers and TaqMan probes were designed in this study based on the conserved regions of the 16S rRNA gene of two pathogens in NCBI.A dual TaqMan fluorescence quantitative PCR method for simultaneous detection of MG and MS was established by optimizing the reaction conditions,and the specificity,sensitivity,repeatability,and reliability of the method were verified.The results showed that this method could specifically amplify MG and MS without cross reactivity with 21 pathogens.The sensitivity experiment results showed that the detection limits of this method for MG and MS were 5.40×10 1 copies/μL and 6.60 × 10 1 copies/μL,and the sensitivity was 10 to 100 times higher than that of known methods.In addition,the re-sults of repetitive experiments showed that the coefficient of variation within and between groups was less than 1％.Compared with the single PCR amplification method in NY/T 553-2015,the positive sample detection coincidence rate,negative sample detection coincidence rate,and total sample detection coincidence rate were all 100.00％,indicating the strong reliability of this method.Using this method to detect 84 suspected Mycoplasma infected chicken samples,the results showed that the MG positivity rate was 32.14％(27/84),the MS positivity rate was 22.62％(19/84),and the positivity rate of samples infected with MG and MS was 16.67％(14/84).Concurrent-ly,182 healthy chicken cloacal swab samples,118 healthy chicken nasal swab samples,and 74 chicken farm environmental samples were detected,and the results showed that all samples were positive for Mycoplasma.The above results indicate that this method can be applied to the detec-tion of various clinical samples.In summary,the method established in this study had the advanta-ges of high specificity,high sensitivity,and good reproducibility.It could be used for clinical differ-ential diagnosis,epidemiological investigation,and pathogen purification of MG and MS infections.

Objective:To explore differences in the clinical and pathological features and postoperative survival after radical resection of biliary tract cancer in different locations such as intrahepatic cholangiocarcinoma,perihilar cholangiocarcinoma,distal cholangiocarcinoma,and gallbladder cancer.Methods:This is a retrospective case series study. The clinical and pathological data of 4 852 patients with biliary tract cancer admitted to the Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xi ′an Jiaotong University from January 2013 to December 2022 were retrospectively analyzed. Among them, 2 110(43.49%%) patients were male and 2 742(56.51%) patients were female,aged from 26 to 88 years with age of (61.3±10.8) years. Observation indicators: (1) The distribution,diagnosis and treatment of patients with biliary tract cancer; (2) Comparison of clinical and pathological features of patients with biliary tract cancer after curative-intent resection; (3) Survival analysis of patients with biliary tract cancer after curative-intent resection; (4) Analysis of effect on adjuvant therapy for patients with biliary tract cancer after curative-intent resection. One-way analysis of variance,Kruskal-Wallis H test and χ 2 test were used for among-group comparisons,respectively. Survival univariate analysis was performed using the Kaplan-Meier method and Log-rank test. Results:Among the 4 852 patients with biliary tract cancer,there were 2 303 cases (47.46%) of gallbladder cancer,952 cases (19.62%) of intrahepatic cholangiocarcinoma,892 cases (18.38%) of perihilar cholangiocarcinoma,and 705 cases(14.53%) of distal cholangiocarcinoma. From the perspective of the year of diagnosis and treatment,the overall number of patients diagnosed and treated for biliary tract cancer has shown an upward trend. From the perspective of diagnosis and treatment,the curative-intent resection rate was 33.37%(1 619/4 852),and the curative-intent resection rate of distal cholangiocarcinoma was higher than that of other biliary tract cancer ( χ2=23.897, P<0.01). Univariate analysis showed that there were statistical differences in gender,age,bile duct stones,total bilirubin at admission,carcinoembryonic antigen,CA19-9,CA125,the degree of pathological differentiation,vascular invasion,microvascular invasion,perineural invasion,surgical margins,pT staging,and pN staging among patients for biliary tract cancer in different locations (all P<0.05). Survival comparison analysis showed that recurrence-free survival and overall survival of patients with gallbladder cancer after curative-intent resection were significantly better than those of intrahepatic cholangiocarcinoma,perihilar cholangiocarcinoma,and distal cholangiocarcinoma ( χ 2=87.780,83.717,both P<0.01). Comparing the postoperative prognosis of patients with biliary tract cancer between the two periods of 2013 to 2017 and 2018 to 2022, the results showed that recurrence-free survival and overall survival of patients with biliary tract cancer from 2018 to 2022 were significantly better than those from 2013 to 2017 ( χ 2=31.202,25.615, both P<0.01),and the proportion of early recurrence and short-term death after curative-intent resection was significantly reduced ( χ 2=21.588,9.623, both P<0.01),with gallbladder cancer being the most significant ( P<0.01). Postoperative adjuvant therapy for patients with biliary tract cancer can effectively prolong recurrence-free survival and overall survival ( χ 2=5.033,11.273,both P<0.05). Conclusions:Gallbladder cancer remains the most common biliary tract cancer with a relatively favorable prognosis after radical resection. There are significant differences in the clinical and pathological features of biliary tract cancer in different locations,and patients with adjuvant therapy effectively improving prognosis.

Objective To explore the risk factors for very early recurrence (VER) after curative-intent resection for gallbladder cancer (GBC) patients and construct prediction models for VER based on various machine learning (ML) algorithms. Methods A retrospective study was conducted on 329 GBC patients who underwent curative-intent surgery at our hospital between January 2016 and December 2020. Risk factors for VER were identified, and prediction models were constructed, validated and compared with multiple ML algorithms[logistic regression (LR), support vector machine (SVM), naive Bayes (NB), random forest (RF), light gradient boosting machine (LGB), and extreme gradient boosting (XGB)]based on independent associated factors for VER. Results Among the 329 patients who underwent curative-intent resection in patients with GBC, 162 (49.2%) patients experienced recurrence, including 69 (42.6%) with VER（<6 months） and 93 (57.4%) with non-VER（≥6 months）. Survival analysis showed that patients with VER had significantly worse median overall survival compared to those with non-VER (6 months vs. not arrived，χ2=398.2, P<0.001). Univariate analysis showed that carcinoembryonic antigen (CEA), carbohydrate antigen (CA)19-9, CA-125, tumor differentiation, pathological type, liver involvement, vascular invasion, perineural invasion, TNM stage, T stage and N stage were risk factors of VER (P<0.05), whereas adjuvant chemotherapy was protective factor (P<0.05). Multivariate analysis confirmed CA-125, tumor differentiation, pathological type, vascular invasion and N stage as independent risk factors (P<0.05), whereas adjuvant chemotherapy was independent protective factor (P<0.05). XGB model achieved the best performance with an area under curve (AUC) of 0.841 and an accuracy (ACC) of 83.0% in the validation set. Shapley additive explanations (SHAP) bar plots highlighted tumor differentiation, N stage, pathological type of tumor, and CA-125 the top four features contributing to the model, each positively influencing the predicted probability of VER. Conclusions CA-125, tumor differentiation, pathological type, vascular invasion, N stage and adjuvant chemotherapy are independent factors associated with VER of GBC following curative-intent resection. ML-based prediction models incorporating these factors have the potential to some extent to effectively identify high-risk patients, providing a valuable reference for VER surveillance in GBC.

To rapidly detect and differentiate Mycoplasma gallisepticum(MG)and Mycoplasma synovialis(MS),two sets of specific primers and TaqMan probes were designed in this study based on the conserved regions of the 16S rRNA gene of two pathogens in NCBI.A dual TaqMan fluorescence quantitative PCR method for simultaneous detection of MG and MS was established by optimizing the reaction conditions,and the specificity,sensitivity,repeatability,and reliability of the method were verified.The results showed that this method could specifically amplify MG and MS without cross reactivity with 21 pathogens.The sensitivity experiment results showed that the detection limits of this method for MG and MS were 5.40×10 1 copies/μL and 6.60 × 10 1 copies/μL,and the sensitivity was 10 to 100 times higher than that of known methods.In addition,the re-sults of repetitive experiments showed that the coefficient of variation within and between groups was less than 1％.Compared with the single PCR amplification method in NY/T 553-2015,the positive sample detection coincidence rate,negative sample detection coincidence rate,and total sample detection coincidence rate were all 100.00％,indicating the strong reliability of this method.Using this method to detect 84 suspected Mycoplasma infected chicken samples,the results showed that the MG positivity rate was 32.14％(27/84),the MS positivity rate was 22.62％(19/84),and the positivity rate of samples infected with MG and MS was 16.67％(14/84).Concurrent-ly,182 healthy chicken cloacal swab samples,118 healthy chicken nasal swab samples,and 74 chicken farm environmental samples were detected,and the results showed that all samples were positive for Mycoplasma.The above results indicate that this method can be applied to the detec-tion of various clinical samples.In summary,the method established in this study had the advanta-ges of high specificity,high sensitivity,and good reproducibility.It could be used for clinical differ-ential diagnosis,epidemiological investigation,and pathogen purification of MG and MS infections.

Objective:To explore differences in the clinical and pathological features and postoperative survival after radical resection of biliary tract cancer in different locations such as intrahepatic cholangiocarcinoma,perihilar cholangiocarcinoma,distal cholangiocarcinoma,and gallbladder cancer.Methods:This is a retrospective case series study. The clinical and pathological data of 4 852 patients with biliary tract cancer admitted to the Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xi ′an Jiaotong University from January 2013 to December 2022 were retrospectively analyzed. Among them, 2 110(43.49%%) patients were male and 2 742(56.51%) patients were female,aged from 26 to 88 years with age of (61.3±10.8) years. Observation indicators: (1) The distribution,diagnosis and treatment of patients with biliary tract cancer; (2) Comparison of clinical and pathological features of patients with biliary tract cancer after curative-intent resection; (3) Survival analysis of patients with biliary tract cancer after curative-intent resection; (4) Analysis of effect on adjuvant therapy for patients with biliary tract cancer after curative-intent resection. One-way analysis of variance,Kruskal-Wallis H test and χ 2 test were used for among-group comparisons,respectively. Survival univariate analysis was performed using the Kaplan-Meier method and Log-rank test. Results:Among the 4 852 patients with biliary tract cancer,there were 2 303 cases (47.46%) of gallbladder cancer,952 cases (19.62%) of intrahepatic cholangiocarcinoma,892 cases (18.38%) of perihilar cholangiocarcinoma,and 705 cases(14.53%) of distal cholangiocarcinoma. From the perspective of the year of diagnosis and treatment,the overall number of patients diagnosed and treated for biliary tract cancer has shown an upward trend. From the perspective of diagnosis and treatment,the curative-intent resection rate was 33.37%(1 619/4 852),and the curative-intent resection rate of distal cholangiocarcinoma was higher than that of other biliary tract cancer ( χ2=23.897, P<0.01). Univariate analysis showed that there were statistical differences in gender,age,bile duct stones,total bilirubin at admission,carcinoembryonic antigen,CA19-9,CA125,the degree of pathological differentiation,vascular invasion,microvascular invasion,perineural invasion,surgical margins,pT staging,and pN staging among patients for biliary tract cancer in different locations (all P<0.05). Survival comparison analysis showed that recurrence-free survival and overall survival of patients with gallbladder cancer after curative-intent resection were significantly better than those of intrahepatic cholangiocarcinoma,perihilar cholangiocarcinoma,and distal cholangiocarcinoma ( χ 2=87.780,83.717,both P<0.01). Comparing the postoperative prognosis of patients with biliary tract cancer between the two periods of 2013 to 2017 and 2018 to 2022, the results showed that recurrence-free survival and overall survival of patients with biliary tract cancer from 2018 to 2022 were significantly better than those from 2013 to 2017 ( χ 2=31.202,25.615, both P<0.01),and the proportion of early recurrence and short-term death after curative-intent resection was significantly reduced ( χ 2=21.588,9.623, both P<0.01),with gallbladder cancer being the most significant ( P<0.01). Postoperative adjuvant therapy for patients with biliary tract cancer can effectively prolong recurrence-free survival and overall survival ( χ 2=5.033,11.273,both P<0.05). Conclusions:Gallbladder cancer remains the most common biliary tract cancer with a relatively favorable prognosis after radical resection. There are significant differences in the clinical and pathological features of biliary tract cancer in different locations,and patients with adjuvant therapy effectively improving prognosis.

Objective:To investigate the efficacy of liver wedge resection and liver Ⅳb and Ⅴ segmentectomy for T2 gallbladder carcinoma (GBC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 168 patients who underwent radical resection of T2 GBC in The First Affiliated Hospital of Xi′an Jiaotong University from January 2011 to December 2021 were collected. There were 59 males and 109 females, aged (65±10)years. Of 168 patients, there were 112 cases in T2a stage and 56 cases in T2b stage. Of 112 patients in T2a stage, 73 cases underwent liver wedge resection and 39 cases underwent liver Ⅳb and Ⅴ segmentectomy. Of 56 patients in T2b stage, 27 cases underwent liver wedge resection and 29 cases underwent liver Ⅳb and Ⅴ segmen-tectomy. Measurement data with normal distribution were represented as Mean± SD, and measure-ment data with skewed distribution were represented as M(range). Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the Mann-Whitney U test. The Kaplan-Meier method was used to calculate survival rate and draw survival curve, and the Log-rank test was used for survival analysis. The COX proportional risk model was used for univariate and multivariate analyses. Results:(1) Clinical data analysis of patients undergoing different extent of hepatic resection for T2 GBC. There was no significant difference in gender, age, cholecystoli-thiasis, preoperative total bilirubin, carcinoembryonic antigen, CA19-9, CA125, incidental GBC, perineural invasion, microvascular invasion, pathological differentiation, histopathological subtypes, N staging, TNM staging between patients with T2a and T2b GBC who underwent different extent of hepatic resection ( P>0.05). (2) Prognostic analysis of T2 GBC patients undergoing different extent of hepatic resection. The 1-, 3- and 5-year cumulative disease-free survival rates of T2 GBC patients undergoing liver wedge resection were 78.0%, 60.1% and 51.4%, respectively, versus 86.8%, 80.0% and 68.0% of T2 GBC patients undergoing liver Ⅳb and Ⅴ segmentectomy, showing a significant difference between them ( χ2 =5.205, P<0.05). The 1-, 3-, and 5-year cumulative overall survival rates of T2 GBC patients undergoing liver wedge resection were 85.0%, 62.5%, and 55.1%, respectively, versus 92.6%, 81.6%, and 68.8% for T2 GBC patients undergoing liver Ⅳb and Ⅴ segmentectomy, showing a significant difference in cumulative overall survival rate between them ( χ2=4.351, P<0.05). The 1-, 3-, and 5-year cumulative disease-free survival rates of T2b GBC patients undergoing liver wedge resection were 70.4%, 45.9% and 39.2%, respectively, versus 89.7%, 71.3% and 54.0% of T2b GBC patients undergoing liver Ⅳb and Ⅴ segmentectomy, showing a significant difference between them ( χ2=5.047, P<0.05). The 1-, 3-, and 5-year cumulative overall survival rates of T2b GBC patients undergoing liver wedge resection were 81.5%, 53.2%, and 41.0%, respectively, versus 89.7%, 77.0%, and 60.7% of T2b GBC patients undergoing liver Ⅳb and Ⅴ segmentectomy, showing no significant difference in cumulative overall survival rate between them ( χ2=4.014, P<0.05). (3) Analysis of factors influencing prognosis of patients undergoing radical resection for T2 GBC. Results of multivariate analysis showed that CA19-9>39.0 U/mL, perineural invasion, N1 and N2 stage were independent risk factors influencing disease-free survival time of patients undergoing radical resection for T2 GBC ( hazard ratio=2.736, 3.496, 2.638, 17.440, 95% confidence interval as 1.195-6.266, 1.213-10.073, 1.429-4.869, 8.362-36.374, P<0.05). Liver Ⅳb and Ⅴ segmentectomy was an independent protective factor influencing disease-free survival time of patients undergoing radical resection for T2 GBC ( hazard ratio=0.418, 95% confidence interval as 0.230-0.759, P<0.05). CA19-9 >39.0 U/mL, perineural invasion, ⅡB stage, ⅢB stage and ⅣB stage of TNM staging were independent risk factors influencing overall survival time of patients undergoing radical resection for T2 GBC ( hazard ratio=2.740, 3.210, 2.037, 3.439, 24.466, 95% confidence interval as 1.127-6.664, 1.049-9.819, 1.004-4.125, 1.730-6.846, 10.733-55.842, P<0.05). Liver Ⅳb and Ⅴ segmentectomy was an independent protective factor influencing overall survival time of patients undergoing radical resec-tion for T2 GBC ( hazard ratio=0.476, 95% confidence interval as 0.261-0.867, P<0.05). (4) Analysis of postoperative complications in patients undergoing different extent of hepatic resection for T2 GBC. There was no significant difference in postoperative complications of patients with T2a and T2b GBC undergoing liver wedge resection or liver Ⅳb and Ⅴ segmentectomy ( P>0.05). Conclusions:Compared to liver wedge resection, liver Ⅳb and Ⅴ segmentectomy can effectively prolong the disease-free survival overall survival time of T2b GBC patients. There is no significant difference in the major complications. Liver Ⅳb and Ⅴ segmentectomy is an independent protective factor for prognosis of patients undergoing radical resection for T2 GBC.

Objective:To explore the value of a new modified T3 sub-staging for the prognosis evaluation in gallbladder cancer patients.Methods:This is a retrospective case-series study. The clinical data of patients with pathologically confirmed stage T3 gallbladder cancer who were admitted to the Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xi′an Jiaotong University from January 2011 to December 2021 were retrospectively analyzed. A total of 190 patients were enrolled in this study, 67 males and 123 females, with an age ( M(IQR)) of 63(14) years (range:17 to 88 years). The stage T3 was divided into four sub-stages according to the site of tumor invasion: (1) T3a:tumor perforates the serosa,but not invading the liver and one other adjacent structure; (2) T3b:tumor perforates the serosa and invades one other adjacent structure,but not the liver; (3) T3c:tumor perforates the serosa and invades the liver,but not one other adjacent structure; (4) T3d:tumor perforates the serosa,invades the liver and one other adjacent structure. To evaluate the application value of this modified sub-staging,the Kaplan-Meier method was used to draw the survival curve,univariate analysis and multivariate analysis were done using the Log-rank test and Cox proportional hazard model respectively. Results:According to the modified T3 sub-staging method,34 patients (17.9%) were in stage T3a,24 cases(12.6%) were in stage T3b, 97 cases (51.1%) were in stage T3c, and 35 cases (18.4%) were in stage T3d. The median survival time of patients in stages T3a,T3b,T3c and T3d after radical resection was 72.0 months, 32.0 months, 12.0 months and 10.0 months, respectively. The 1-, 3-, and 5-year survival rates of patients in stage T3a, T3b, T3c and T3d were 79.4%, 53.3%, and 53.3%; 79.2%, 44.6%, and 26.0%;49.5%,27.5%,and 18.1%;42.9%,15.9%, and 15.9% ( χ2=18.349, P<0.01),respectively. Univariate analysis showed that gallbladder stones,pathological differentiation,perineural invasion, N stage,postoperative adjuvant therapy and modified T3 substage were factors affecting patient prognosis(all P<0.05). Cox multivariate analysis showed that modified sub-stages with T3c ( HR=2.043, 95% CI:1.176 to 3.549) and T3d( HR=2.419, 95% CI:1.284 to 4.555), accompanied by gallbladder stones ( HR=1.661,95% CI:1.150 to 2.398),pathological differentiation with poorly differentiated( HR=1.709,95% CI:1.198 to 2.438), and the N stage with N1 and N2( HR=1.602, 95% CI:1.090 to 2.355, 2.714, 95% CI: 1.621 to 4.544) were independent prognostic risk factors for patients in stage T3,while postoperative adjuvant chemotherapy( HR=0.351) was a protective factor for prognosis. There was no statistically significant difference in survival between patients with stage T3a and T3b who underwent hepatic wedge resection and liver segment or major resection ( P=0.402). For patients with stage T3c and T3d with liver invasion,the survival difference after hepatic wedge resection and segmental or major resection was statistically significant ( P=0.008). Conclusion:The modified T3 sub-staging system based on the depth and direction of tumor invasion maybe helpful to further stratify the prognosis of patients with gallbladder cancer.

Objective To monitor the antifungal resistance of clinical isolates of Candida spp.in the East China region.Methods MALDI-TOF MS or molecular methods were used to re-identify the strains collected from January 2018 to December 2022.Antifungal susceptibility testing was performed using the broth microdilution method.The susceptibility test results were interpreted according to the breakpoints of 2022 Clinical and Laboratory Standards Institute(CLSI)documents M27 M44s-Ed3 and M57s-Ed4.Results A total of 3 026 strains of Candida were collected,65.33％of which were isolated from sterile body sites,mainly from blood(38.86％)and pleural effusion/ascites(10.21％).The predominant species of Candida were Candida albicans(44.51％),followed by Candida parapsilosis complex(19.46％),Candida tropicalis(13.98％),Candida glabrata(10.34％),and other Candida species(0.79％).Candida albicans showed overall high susceptibility rates to the 10 antifungal drugs tested(the lowest rate being 93.62％).Only 2.97％of the strains showed dose-dependent susceptibility(SDD)to fluconazole.Candida parapsilosis complex had a SDD rate of 2.61％and a resistance rate of 9.42％to fluconazole,and susceptibility rates above 90％to other drugs.Candida glabrata had a SDD rate of 92.01％and a resistance rate of 7.99％to fluconazole,resistance rates of 32.27％and 48.24％to posaconazole and voriconazole non-wild-type strains(NWT),respectively,and susceptibility rates above 90％to other drugs.Candida tropicalis had resistance rates of 29.55％and 26.24％to fluconazole and voriconazole,respectively,resistance rates of 76.60％and 21.99％to posaconazole and echinocandins non-wild-type strains(NWT),and a resistance rate of 2.36％to echinocandins.Conclusions The prevalence and species distribution of Candida spp.in the East China region are consistent with previous domestic and international reports.Candida glabrata exhibits certain degree of resistance to fluconazole,while Candida tropicalis demonstrates higher resistance to triazole drugs.Additionally,echinocandins resistance has emerged in Candida albicans,Candida glabrata,Candida tropicalis,and Candida parapsilosis.

Pancreatobiliary maljunction(PBM)represents a congenital anatomical abnormality of the pancreaticobiliary ductal system,frequently predisposing individuals to recurrent cholangitis and pancreatitis.Accumulating evidence indicates that PBM is a precancerous lesion,and PBM plays an important role in the development and progression of gallbladder cancer(GBC).GBC arising from PBM is designated as PBM-associated GBC.Consequently,early diagnosis and treatment of PBM is paramount in mitigating the risk of GBC.This review outlined the epidemiology and advancements in the diagnosis and management of PBM,along with the clinical features,underlying mechanisms,and therapeutic progressions pertaining to PBM-associated GBC,in order to underscore the clinical significance of early intervention in PBM,so as to reduce the incidence of PBM-associated GBC.