Improving the nitrogen use efficiency (NUE) of Brassica napus is of significant importance for achieving the national goal of zero growth in chemical fertilizer application and ensuring the green development of the rapeseed industry. This study aims to explore the effects of the nitrate transporter gene BnaNRT1.5s on the nitrogen transport and NUE of B. napus, providing excellent genetic resources for the development of nitrogen-efficient B. napus varieties. The spatiotemporal expression of BnaA05.NRT1.5 as a key nitrogen responsive gene was profiled by qRT-PCR at different growth stages and for different tissue samples of B. napus 'Westar'. Subcellular localization was employed to examine its expression pattern in the cells. Additionally, CRISPR/Cas9 was used to create BnaNRT1.5s knockout lines, which were subjected to hydroponic experiments under high nitrogen (12.0 mmol/L) and low nitrogen (0.3 mmol/L) conditions. After the seedlings were cultivated for 21 days, root and shoot samples were collected for weighing, nitrogen content determination, xylem sap nitrate content assessment, and calculation of total nitrogen and NUE. The B. napus nitrate transporter BnaA05.NRT1.5 was localized to the cell membrane. During the seedling and early bolting stages, BnaA05.NRT1.5 was predominantly expressed in roots, while it was highly expressed in old leaves and mature silique skin during the reproductive stage. Compared with the wild type, the mutant BnaNRT1.5s showed significant increases in the dry weight and total nitrogen of seedlings under both high and low nitrogen conditions. Under low nitrogen conditions, NUE in the roots of BnaNRT1.5s significantly improved. Notably, under both high and low nitrogen conditions, the nitrate content in the shoots of BnaNRT1.5s decreased significantly, while that in the roots increased significantly, resulting in a significantly decreased shoot-to-root nitrate content ratio. BnaNRT1.5s is involved in regulating the transport of nitrate from the roots to the shoots, and its mutation enhances nitrogen absorption and utilization in B. napus seedlings, promoting seedling growth. This study not only provides references for understanding the physiological and molecular mechanisms by which BnaNRT1.5s regulates NUE but also offers valuable genetic resources for improving NUE in B. napus.
Brassica napus/genetics* ; Anion Transport Proteins/metabolism* ; Nitrogen/metabolism* ; Nitrate Transporters ; Plant Proteins/metabolism* ; Nitrates/metabolism* ; Gene Expression Regulation, Plant ; Biological Transport

Objective To explore the mediating effect of sleep duration on the relationship between qi stagnation constitution and anxiety/depression in junior high school students.Methods From March 20 to March 31,2020,a total of 10,325 students from 6 junior high schools in Henan,Liaoning,Shaanxi,Shandong,and Fujian provinces were sampled by cluster sampling.Firstly,the general conditions(age,gender,grade,hometown,whether they are an only child)and sleep duration were collected,and their qi stagnation constitution,depression and anxiety status were evaluated by using the Qi Stagnation Subscale in the"TCM Constitution Classification and Judgment Scale",the Patient Health Questionnaire(PHQ-9)and the Generalized Anxiety Disorder(GAD-7).Secondly,Spearman correlation analysis was used to explore the correlation between qi stagnation constitution,sleep duration,and anxiety/depression.Then,linear regression analysis and mediating effect test were used to explore the relationship between qi stagnation constitution and anxiety/depression in junior high school students,and the mediating effect of sleep duration was analyzed,and the results were verified.Results(ⅰ)The detection rate of anxiety in non-only children was higher than that of only children(x2=11.198,P=0.001);The detection rate of anxiety(x2=106.967,P＜0.001)or depression(x2=84.692,P＜0.001)was higher among senior students than those in lower grades,while those of girls with anxiety(x2=100.441,P＜0.001)or depression(x2=71.418,P＜0.001)were higher than those of boys.The detection rates of anxiety(x2=1 092.298,P＜0.001)and depression(x2=866.740,P＜0.001)in qi stagnation constitution students were higher than those of non-qi stagnation constitution students,and the detection rates of anxiety(x2=1 076.716,P＜0.001)and depression(x2=1 099.725,P＜0.001)in students whose sleep duration less than 8 h were higher than those of whose sleep duration more than 8 h,and the shorter the sleep duration,the higher the detection rate.(ⅱ)Qi stagnation constitution positively predicted anxiety/depression(β=0.679,P＜0.001;β=0.718,P＜0.001),and sleep duration had a negative predictive effect on anxiety/depression(β=-0.403,P＜0.001;β=-0.439,P＜0.001).(ⅲ)Sleep duration played a partial mediating role in the relationship between qi stagnation constitution and anxiety/depression in junior high school students,and the mediating effect accounted for 13.40％and 13.79％of the total effect,respectively.Conclusion Although qi stagnation constitution mainly affects anxiety/depression through direct effect,it can still be partially indirectly realized through the mediating variable of sleep duration,that is,the shorter the sleep duration,the higher the risk of anxiety/depression in junior high school students.Therefore,the mental health problems of junior high school students can be prevented and improved by regulating qi stagnation constitution and ensuring adequate sleep duration.

Objective:To establish and verify the prediction model of benign or malignant of solitary pulmonary nodules (SPNs≤20 mm) based on artificial intelligence.Methods:Totally 338 SPNs (≤20 mm) from 279 patients, confirmed by operation and pathology, were selected in Zhongshan Hospital Xiamen University from November 2018 to May 2020. Clinical data (age, gender, smoking history, individual and family history of malignancy), image features (maximum diameter, minimum diameter, solid proportion, volume, lobulation sign, burr sign, vacuole sign, cavity sign, pleural indentation sign, and radiomic features (maximum CT value, minimum CT value, average CT value, median CT value, CT value standard deviation, skewness, peak, energy, entropy) were analyzed retrospectively. All the data of patients were randomly divided into training set (271 SPNs) and test set (67 SPNs). In the training set, the clinical, image features and radiomic features were first selected by the least absolute shrinkage and selection operator (LASSO) regression, then the independent risk factors of SPN (≤20 mm) were screened out by multi-variate logistic regression analysis, and the nomogram prediction models were constructed. Finally, the data of test set were used to verify the prediction model by the ROC curve and calibration curve (CC).Results:In the training set of 271 SPNs, 81 SPNs were benign and 190 malignant. After analysis of LASSO regression and multi-factor logistics regression, the independent predictors of benign or malignant SPN were age, gender, largest diameter, vacuole sign and solid proportion. The prediction model was P=e x/(1+e x), x=-2.583+0.027×age+1.519×gender+0.127×maximum diameter-2.132×solid proportion+1.720×vacuole sign. The results of the model showed that the area under curve (AUC) of ROC was 0.850, and the sensitivity was 73.7%, specificity was 82.7% and accuracy was 82.3%. In the test set of 67 SPNs, 22 SPNs were benign and 45 malignant. The results showed that the AUC of ROC was 0.882, and the sensitivity was 82.2%, specificity was 81.8% and accuracy was 85.1%. The calibration nomogram of prediction model showed that CC from training set or test set well coincided with its individual ideal curve ( Ptraining=0.688, Ptest=0.618). Conclusion:Prediction model of benign or malignant SPN ≤20 mm is established based on AI; it can obtain the prediction probability and has good diagnostic efficiency.

Objective This research is aimed to investigate the efficacy and toxicity of icotinib for lung adenocarcinoma pa-tients with poor performance status and unknown EGFR gene status .Methods A total of 27 lung adenocarcinom patients with poor Eastern Cooperative Oncology Group-Performance status(ECOG-PS) and unknown EGFR gene status referred to Chongqing Canc-er Institute from August 2012 to August 2014 were analyzed .Icotinib (125 mg) was orally administered three times per day .Asess the efficacy and adverse reaction ,calculate survival rates .Results Among the 27 patients ,the objective response rate(ORR) and disease control rates(DCR) were 29 .6% and 81 .5% ,respectively .The median progression free survival time was 6 .0 months .A to-tal of 70 .4% of patients had an significant improvment in ECOG-PS scores ,following icotinib treatment (Z= - 2 .157 ,P= 0 .031) . Fatigue ,anorexia and diarrhea were the most frequent adverse reaction ,which defined as grade 1 to 2 rashes .Conclusion Lung ade-nocarcinoma patients with poor performance status and unknown EGFR gene status may benefit from icotinib therapy ,and patients were tolerated well .

Objective To compare the efficiency of laparoscopic cholecystectomy (LC) and radical cholecystectomy (RC) in the patients with early gallbladder carcinoma. Methods From January 2001 to December 2008, 63 patients who had underwent LC(28 cases) or RC(35 cases) were analyzed retrospectively. The survival and recurrence rate after operation and complication were compared. Results After treatment, the complication in the LC group was less than that in the RC group (7. 1% vs. 48. 6%, χ2 = 12. 675 ,P ＜0.01). In the RC group,the 1,3,5 year overall cumulative survival rate of Nevein phase Ⅰ were all 100%, and 76. 2%, 42. 9%, and 23. 8% respectively in the phase Ⅱ. The overall cumulative survival rate of LC cases were 81.8%, 54. 5%,27.3% in phase Ⅰ and 17.6% ,5.9% ,0.0% in phase Ⅱ, respectively. The survival curves between the two groups were significantly diffierent(phase Ⅰ χ2 =20.74,P＜0.001; phase Ⅱ χ2 = 11.35,P＜0.001) and the survival rate in the RC group was higher than that in the LC group. The 1,3,5 year cumulative recurrence rates in the RC were 20.0%, 48. 6%, 68. 6%, respectively, which is significantly lower than those in the LC group (60.7%,85.7% and92.9%, respectively)(χ2 =10.30,P=0.0013).Conclusions RC is one of the first choices for early gallbladder carcinoma treatment, and will decrease the recurrence rate and improve the long-term life quality.