1.BnaNRT1.5s mediates nitrate transporter to regulate nitrogen use efficiency in Brassica napus.
Shilong CHEN ; Lei YAO ; Rumeng WANG ; Jian ZENG ; Jianghe LI ; Shiyao CUI ; Xu WANG ; Haixing SONG ; Zhenhua ZHANG ; Pan GONG
Chinese Journal of Biotechnology 2025;41(7):2954-2965
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*
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Anion Transport Proteins/metabolism*
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Nitrogen/metabolism*
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Nitrate Transporters
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Plant Proteins/metabolism*
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Nitrates/metabolism*
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Gene Expression Regulation, Plant
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Biological Transport
2.Application value of dual amplification method for nucleic acid detection of seven respiratory pathogens by throat swab samples in diagnosis of acute upper respiratory tract infections in children
Yongqing ZHOU ; Jianghe WANG ; Hengyan LIN ; Kaiqi YANG ; Nan RAO ; Man WANG ; Hongjuan CHU
Journal of Clinical Medicine in Practice 2025;29(3):114-117
Objective To compare the diagnostic values of different detection methods for respir-atory pathogens in children with acute respiratory infections.Methods A total of 862 children with a-cute respiratory infections were enrolled,and their throat swab samples were tested for seven common respiratory pathogens by the dual amplification method and a self-built nucleic acid detection system.For samples with inconsistent results between the two methods,nested polymerase chain reaction(PCR)was performed for verification.Results The positive detection rate of the dual amplification method was 57.75%,which was significantly higher than 30.14%of the self-built nucleic acid detec-tion system,and the detection rate of mixed infections was 10.14%,which was also significantly high-er than 1.97%of the self-built nucleic acid detection system(P<0.05).The sensitivity of the dual amplification method was 91.63%,which was significantly higher than 72.61%of the self-built nu-cleic acid detection system,and the specificity was 92.31%,which was also significantly higher than 75.62%of the self-built nucleic acid detection system(P<0.05).Conclusion The dual amplifica-tion method can simultaneously detect the ribonucleic acid of seven respiratory pathogens with high sensitivity and specificity,demonstrating significant clinical application value.
3.Relationship between qi stagnation constitution and anxiety/depression in 10,325 junior high school students,and in which the mediating effect of sleep duration
Mengshu WANG ; Yi LI ; Yun DU ; Jinhe DAI ; Jianghe CHEN ; Miao QU
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1457-1465
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.
4.Establishment and verification of prediction model for benign or malignant of≤20 mm solitary pulmonary nodules
Hua ZHONG ; Anqi LI ; Jianghe KANG ; Jin′an WANG ; Shaoyin DUAN
Chinese Journal of Radiology 2021;55(7):745-750
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.
5.Targeting castration-resistant prostate cancer with a novel ROR
Jianwei ZHENG ; Junfeng WANG ; Qian WANG ; Hongye ZOU ; Hong WANG ; Zhenhua ZHANG ; Jianghe CHEN ; Qianqian WANG ; Panxia WANG ; Yueshan ZHAO ; Jing LU ; Xiaolei ZHANG ; Songtao XIANG ; Haibin WANG ; Jinping LEI ; Hong-Wu CHEN ; Peiqing LIU ; Yonghong LIU ; Fanghai HAN ; Junjian WANG
Acta Pharmaceutica Sinica B 2020;10(12):2313-2322
Prostate cancer (PCa) patients who progress to metastatic castration-resistant PCa (mCRPC) mostly have poor outcomes due to the lack of effective therapies. Our recent study established the orphan nuclear receptor ROR
6.Detection of transgenic components in animal feeds on Shanxi markets.
Jianqin YUAN ; Hong CHANG ; Jianghe ZHAO ; Zhongwei TANG ; Zongyong SHI ; Jundong WANG
Chinese Journal of Biotechnology 2016;32(11):1576-1589
To assess the presence of genetically modified (GM) maize and soybean in a range of commercialized feed in Shanxi province of China in 2015, improved hexadecyltrimethy ammonium bromide (CTAB) method was used to extract DNA. The screening of packed feeds was carried out by qualitative PCR. Then positive feeds were unpacked and detected by the CaMV 35S promoter, NOS terminator, zSSIIb, Lectin and CryIA (b) genes. The identified maize and soybean events were confirmed by event-specific MON810 and GTS40-3-2. Results showed that 83.3% of the feeds was tested positive for GMOs, in which positive rates of maize, soybean, pig and layer feeds were 6.67%, 100%, 93.3% and 73.3%, respectively. The results of real-time PCR were consistent with qualitative PCR. These results indicated that commercialized GM feed had a wide positive product scope in Shanxi province of China. Further studies are necessary to study effects of feeding livestock and poultry with feed containing GM ingredients on animals and their products.
7.Analysis of exogenous gene and protein digestion and absorption of SD rats (Rattus norvegicus) fed roundup ready soybean meal.
Jianqin YUAN ; Hong CHANG ; Jianghe ZHAO ; Zongyong SHI ; Jundong WANG
Chinese Journal of Biotechnology 2016;32(5):657-668
Metabolism and deposition of exogenous gene and protein from transgenic glyphosate herbicide-tolerant soybean meal in SD rats were studied in the experiment. The transgenic soybean GTS40-3-2 meal and its non-transgenic counterpart (parent A5403) were fed to the generation and the second generation Sprague-Dawley (SD) rats (Rattus norvegicus). The study added the genetically modified (GM) soybean meal and its non-transgenic control soybean meal (parent A5403) in a ratio of 20% respectively to the feeds. By using qualitative, quantitative PCR and ELISA methods to detect transgenic soybean residues of metabolism ingredients in rats, the safety and influence of GM soybean were evaluated. The results showed that the intestinal fecal and cecum contents of rats were detected with residues of GM ingredients, intestinal flora and organs were not found related genes and protein. These results indicated that transgenic glyphosate herbicide-tolerant soybean GTS40-3-2 meal was as safe as its non-GM soybean meal in long-term feeding study.
Animal Feed
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Animal Nutritional Physiological Phenomena
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Animals
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Digestion
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Glycine
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analogs & derivatives
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Herbicide Resistance
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Herbicides
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Plants, Genetically Modified
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Proteolysis
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Rats
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Rats, Sprague-Dawley
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Soybean Proteins
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Soybeans
8.Clinical effects of icotinib on lung denocarcinoma patients with unknown EGFR gene status and poor performance status
Xiaohui JI ; Lumi HUANG ; Chunmei WANG ; Jianghe SHAO ; Donglin WANG ; Dairong LI
Chongqing Medicine 2016;(2):189-191,195
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 .

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