1.Mapping of QTL associated with rice cooking quality and candidate gene analysis.
Qiaona LE ; Ziwen HUANG ; Ruohui DAI ; Sanfeng LI ; Mengjia LI ; Yuan FANG ; Yuexing WANG ; Yuchun RAO
Chinese Journal of Biotechnology 2024;40(1):122-136
Excavating the quantitative trait locus (QTL) associated with rice cooking quality, analyzing candidate genes, and improving cooking quality-associated traits of rice varieties by genetic breeding can effectively improve the taste of rice. In this study, we used the indica rice HZ, the japonica rice Nekken2 and 120 recombinant inbred lines (RILs) populations constructed from them as experimental materials to measure the gelatinization temperature (GT), gel consistency (GC) and amylose content (AC) of rice at the maturity stage. We combined the high-density genetic map for QTL mapping. A total of 26 QTLs associated with rice cooking quality (1 QTL associated with GT, 13 QTLs associated with GC, and 12 QTLs associated with AC) were detected, among which the highest likelihood of odd (LOD) value reached 30.24. The expression levels of candidate genes in the localization interval were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), and it was found that the expression levels of six genes were significantly different from that in parents. It was speculated that the high expression of LOC_Os04g20270 and LOC_Os11g40100 may greatly increase the GC of rice, while the high expression of LOC_Os01g04920 and LOC_Os02g17500 and the low expression of LOC_Os03g02650 and LOC_Os05g25840 may reduce the AC. The results lay a molecular foundation for the cultivation of new high-quality rice varieties, and provide important genetic resources for revealing the molecular regulation mechanism of rice cooking quality.
Quantitative Trait Loci
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Oryza/genetics*
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Plant Breeding
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Cooking
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Genetic Association Studies
2.Exploring the innovative talents training mode in new era.
Li MA ; Siyi SHEN ; Yuchun RAO
Chinese Journal of Biotechnology 2024;40(1):292-303
Innovation is an important way to promote economic development and social progress. Recent years have seen rapid development of biological sciences. In response to social demands and the needs for developing an innovative country, fostering innovative talents in the field of biosciences has become a significant initiative supported by national policies and the needs from talent market. Taking the innovative talent training mode implemented by Zhejiang Normal University in the field of biological sciences as an example, this paper comprehensively introduces several key aspects of the mode. This includes establishing a mentorship system as the foundation, carrying out curriculum reform through project competitions and practical platforms, and promoting synergy among industry, academia, and research in talent training. This training mode has achieved positive results in practice, promoting the training of outstanding innovative talents in biological science majors, and may facilitate the reform of talent training in similar majors.
Humans
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Biological Science Disciplines
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Industry
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Policy
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Universities
3.Application of CRISPR-Cas9 gene editing technology in crop breeding.
Wenjing YIN ; Zhengai CHEN ; Jiahui HUANG ; Hanfei YE ; Tao LU ; Mei LU ; Yuchun RAO
Chinese Journal of Biotechnology 2023;39(2):399-424
The CRISPR-Cas9 system is composed of a clustered regularly interspaced short palindromic repeat (CRISPR) and its associated proteins, which are widely present in bacteria and archaea, serving as a specific immune protection against viral and phage secondary infections. CRISPR-Cas9 technology is the third generation of targeted genome editing technologies following zinc finger nucleases (ZFNs) and transcription activator like effector nucleases (TALENs). The CRISPR-Cas9 technology is now widely used in various fields. Firstly, this article introduces the generation, working mechanism and advantages of CRISPR-Cas9 technology; secondly, it reviews the applications of CRISPR-Cas9 technology in gene knockout, gene knock-in, gene regulation and genome in breeding and domestication of important food crops such as rice, wheat, maize, soybean and potato. Finally, the article summarizes the current problems and challenges encountered by CRISPR-Cas9 technology and prospects future development and application of CRISPR-Cas9 technology.
Gene Editing
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CRISPR-Cas Systems/genetics*
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Plant Breeding
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Crops, Agricultural/genetics*
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Technology
4.Construction and preliminary validation of a risk prediction model for the recurrence of diabetic foot ulcer in diabetic patients
Qingjiao GUO ; Jing OUYANG ; Jiaqin RAO ; Yizhi ZHANG ; Lihong YU ; Wanying XU ; Jinhua LONG ; Xiuhua GAO ; Xiaoyan WU ; Ying GU
Chinese Journal of Burns 2023;39(12):1149-1157
Objective:To develop a risk prediction model for the recurrence of diabetic foot ulcer (DFU) in diabetic patients and primarily validate its predictive value.Methods:Meta-analysis combined with retrospective cohort study was conducted. The Chinese and English papers on risk factors related to DFU recurrence publicly published in China Biology Medicine disc, China National Knowledge Infrastructure, Wanfang Database, VIP Database, and PubMed, Embase, Cochrane Library, and Web of Science, and the search time was from the establishment date of each database until March 31 st, 2022. The papers were screened and evaluated, the data were extracted, a meta-analysis was performed using RevMan 5.4.1 statistical software to screen risk factors for DFU recurrence, and Egger's linear regression was used to assess the publication bias of the study results. Risk factors for DFU recurrence mentioned in ≥3 studies and with statistically significant differences in the meta-analysis were selected as the independent variables to develop a logistic regression model for risk prediction of DFU recurrence. The medical records of 101 patients with DFU who met the inclusion criteria and were admitted to Affiliated Hospital of Guizhou Medical University from January 2019 to June 2022 were collected. There were 69 males and 32 females, aged (63±14) years. The receiver operating characteristic (ROC) curve of the predictive performance of the above constructed predictive model for DFU recurrence was drawn, and the area under the ROC curve, maximum Youden index, and sensitivity and specificity at the point were calculated. Dataset including data of 8 risk factors for DFU recurrence and the DFU recurrence rates of 10 000 cases was simulated using RStudio software and a scatter plot was drawn to determine two probabilities for risk division of DFU recurrence. Using the β coefficients corresponding to 8 DFU recurrence risk factors ×10 and taking the integer as the score of coefficient weight of each risk factor, the total score was obtained by summing up, and the cutoff scores for risk level division were calculated based on the total score × two probabilities for risk division of DFU recurrence. Results:Finally, 20 papers were included, including 3 case-control studies and 17 cohort studies, with a total of 4 238 cases and DFU recurrence rate of 22.7% to 71.2%. Meta-analysis showed that glycosylated hemoglobin >7.5% and with plantar ulcer, diabetic peripheral neuropathy, diabetic peripheral vascular disease, smoking, osteomyelitis, history of amputation/toe amputation, and multidrug-resistant bacterial infection were risk factors for the recurrence of DFU (with odds ratios of 3.27, 3.66, 4.05, 3.94, 1.98, 7.17, 11.96, 3.61, 95% confidence intervals of 2.79-3.84, 2.06-6.50, 2.50-6.58, 2.65-5.84, 1.65-2.38, 2.29-22.47, 4.60-31.14, 3.13-4.17, respectively, P<0.05). There were no statistically significant differences in publication biases of diabetic peripheral neuropathy, diabetic peripheral vascular disease, glycosylated hemoglobin >7.5%, plantar ulcer, smoking, multidrug-resistant bacterial infection, or osteomyelitis ( P>0.05), but there was a statistically significant difference in the publication bias of amputation/toe amputation ( t=-30.39, P<0.05). The area under the ROC curve of the predictive model was 0.81 (with 95% confidence interval of 0.71-0.91) and the maximum Youden index was 0.59, at which the sensitivity was 72% and the specificity was 86%. Ultimately, 29.0% and 44.8% were identified respectively as the cutoff for dividing the probability of low risk and medium risk, and medium risk and high risk for DFU recurrence, while the corresponding total scores of low, medium, and high risks of DFU recurrence were <37, 37-57, and 58-118, respectively. Conclusions:Eight risk factors for DFU recurrence are screened through meta-analysis and the risk prediction model for DFU recurrence is developed, which has moderate predictive accuracy and can provide guidance for healthcare workers to take interventions for patient with DFU recurrence risk.
5.Relationship between the status of BRAF V600 and EGFR gene mutation and clinicopathological features in patients with non-small cell lung cancer
Yiming HAN ; Lan RAO ; Li DING ; Jie ZHENG ; Wan YANG ; Jinhua SHEN ; Xianjin ZOU
Chinese Journal of Clinical and Experimental Pathology 2017;33(4):375-378
Purpose To study the status of BRAF V600 and EGFR mutations in patients with non-small cell lung cancer (NSCLC) and to examine the relations between them.Methods BRAF V600 and EGFR mutations were detected with DNA sequencing.The relationship between BRAF V600,EGFR mutations and the clinicopathological features were analyzed.Results BRAF V600 mutations were detected in 11 (7.5%) of the 146 specimens.BRAF V600 mutations were found morelfrequently in non-smokers (P =0.045).There were no significant differences in age,gender,histological subtype and differentiation between patients with and without BRAF V600 mutations (P > 0.05).EGFR mutations were detected in 68 (46.6%) of the 146 specimens.EGFR mutations were found more frequently in women,non-smokers and adenocarcinoma (P < 0.05).Four tumors with BRAF V600 mutations (three V600 and one V600D) showed concomitant EGFR mutations (two DEL and two L858R).Conclusion BRAF V600 mutations in patients with NSCLC are found more frequently in non-smokers.There are no significant differences in age,gender,histological subtype and differentiation between patients with and without BRAF mutations.
6.Analysis on spatial distribution of tuberculosis in China, 2012-2014
Jin FAN ; Huaxiang RAO ; Peng WU ; Jie ZHANG ; Yanping WU ; Jinhua PAN ; Wenhan LI ; Lixia QIU
Chinese Journal of Epidemiology 2017;38(7):926-930
Objective To analyze the spatial distribution of the incidence of tuberculosis (TB)in China from 2012 to 2014 and provide evidence for the prevention and control of TB.Methods The database of TB in China from 2012 to 2014 was established by using geographical information system,the spatial distribution map was drawn,trend analysis and spatial autocorrelation analysis were conducted to explore the spatial distribution pattern of TB and identify hot areas.Results The trend surface analysis showed that the incidence of TB decreased gradually from the west to the east in China,and the U type curve could reflect the TB distribution from the south to the north;Global spatial autocorrelation analysis showed the 2012-2014 global Moran's I were 0.366,0.364 and 0.358(P<0.01),suggesting that the incidence of TB had a spatial clustering in China;Local Getis-OrdGi spatial autocorrelation analysis by ArcGIS software showed that there was 11 cluster areas,3 high incidence areas (Xinjiang,Tibet,Qinghai) and 8 low incidence areas (Beijing,Tianjin,Shanghai,Hebei,Inner Mongolia,Shanxi,Shandong,Jiangsu).Conclusion The incidence of TB had obviously spatial clustering characteristic,the areas at high risk were mainly in the northwestern and plateau area in China.
7.EGFR gene mutation status, ERCC1 and Ki-67 protein expression in non-small cell lung cancer and relationship with clinicopathologic features
Yiming HAN ; Jie ZHENG ; Yunhui JIANG ; Jinhua SHEN ; Lan RAO ; Wanting FAN ; Junhua XIE ; Xianjin ZOU
Chinese Journal of Clinical and Experimental Pathology 2015;(7):759-763
Purpose To study the status of EGFR mutations and the expression of excision repair cross-complementation group 1 ( ER-CC1) and Ki-67 protein in patients with non-small cell lung cancer (NSCLC) and to examine the relationship between their expression and clinicopathologic features. Methods EGFR mutations were analyzed with DNA sequencing, and the expression of ERCC1 and Ki-67 protein was examined by immunohistochemistry EnVision. The relationship of EGFR mutations with the expression of ERCC1and Ki-67 and the clinicopathological features were analyzed. Results EGFR mutations were detected in 143 (143/291, 49. 1%) of the 291 specimens. EGFR mutations were found more frequently in women, non-smokers and adenocarcinoma. The difference of EGFR muta-tion rate between the histological subtypes according to the IASLC/ATS/ERS classification of lung adenocarcinoma was significantly ( P=0. 008). The mean tumor diameter was smaller in patients with EGFR mutations than in those with wild-type EGFR (P=0. 020). EGFR mutations were not related to age, lymph node metastasis. However, EGFR mutations were not related to the expression of ER-CC1 and Ki-67 protein (P>0. 050). Conclusions EGFR mutation is closely linked to several clinicopathological factors, such as gender, differentiation, and histological subtype. There is heterogeneity of EGFR mutation in patients with NSCLC. EGFR mutations were not related to the expression of ERCC1 and Ki-67 protein.
8.Analysis of risk factors related to the pathogenesis of urolithiasis of migrant workers in Zhongshan City
Jinhua HE ; Shan HUO ; Huiping RAO ; Ruiwen HUANG ; Bin YANG ; Chunchun WEI
Chinese Journal of Primary Medicine and Pharmacy 2013;20(15):2266-2268
Objective To explore the risk factors of urolithiasis incidence of migrant workers in Zhongshan,in order to provide the scientific evidence for preventing urolithiasis.Methods 1 630 workshop migrant workers and 1630 office white collar workers were selected as the subjects of this study.The questionnaire was conducted according to the age,gender,genetic factors,occupation,past illness factors,living environment,work environment factors,diet habit,the habit of drinking water survey,B ultrasound examination,the detection of blood uric acid.Results The incidence rate of urolithiasis was 6.53%,the incidence rate of workshop migrant workers was 8.28%,which was significantly higher than that of white collar 4.72% (x2 =16.972,P <0.01).Conclusion Work environment,eating habits,drinking habits,smoking and sweating are related risk factors of urolithiasis.
9.Determination of evodiamine by high performance liquid chromatography-tandem mass spectrometry and pharmacokinetic studies in rats
Jihua XU ; Wenying LIU ; Feng ZHENG ; Di SUN ; Qian YANG ; Jinhua RAO
Chinese Journal of Clinical Pharmacology and Therapeutics 2007;12(4):427-433
AIM: To establish an LC-MS/MS method for determination of evodiamine concentration in rat plasma and to study its pharmacokinetic profile in rats. METHODS: Six rats were administrated (i.g.) evodiamine at the dose of 100 mg/kg. Blood samples were collected from eye socket. Evodiamine concentration in rat plasma was determined by LC-MS/MS method. The pharmacokinetic parameters were calculated using DAS program. RESULTS: A good linear relationship was obtained in the concentration range (0.2-50.0 ng/mL) studied (r2=0.9997). Average recoveries ranged from 96.12% to 99.46%. Intra-and inter-day relative standard deviations were 4.61%-13.51% and 5.65%-11.49%, respectively. The main pharmacokinetic parameters of evodiamine were as follows: Cmax (5.3±1.5) ng/mL; tmax (22±8) min; t1/2 (451±176) min. CONCLUSION: A selective and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the quantification of evodiamine in rat plasma is developed and validated. This method is successfully applied for the pharmacokinetic studies of evodiamine in rats.
10.A rapid and sensitive method for determination of escitalopram in human plasma and its application in pharmacokinetic study by liquid chromatography-tandem mass spectrometry
Qian YANG ; Wenying LIU ; Feng ZHENG ; Jihua XU ; Jinhua RAO ; Di SUN ; Shu GAO
Chinese Journal of Clinical Pharmacology and Therapeutics 2006;11(10):1148-1153
AIM: To determine the concentration of escitalopram in human plasma by HPLC-MS/MS and investigate the pharmacokinetics of escitalopram. METH-ODS: The method involved protein precipitation with methanol. The chromatographic separation was achieved within 6.0 min by using methanol-water with 15 mmol·L-1 ammonium acetate-formic acid (72:28:O.1, v/v/v) as mobile phase and a Lichrospher CN 150 mm×4.6 mm analytical column. The analytes were detected using an electrospray ionization tandem mass spectrometry in SRM mode. Detection of the ions was performed by monitoring the transitions of m/z 325.0 to 234.0 for escitalopram and m/z 409.1 to 238.1 for amlodipine (intemal standard), respectively. RESULTS:The standard curve was linear ( r = 0. 999) over the concentration range of 0.20 - 50.00 ng· ml- 1. Accuracy and precision were below the acceptance limits of 15%. The recoveries of escitalopram ranged from 96.0% to 103.6%. The lower limit of quantification for escitalopram was 0.20 ng· ml-1 using 200 μl plasma sample.The pharmacokinetic parameters of escitalopram after a single oral dosing of escitalopram oxalate tablet (10 rog)to ten healthy male volunteers were achieved. The Cmax, Tmax, AUC0-t, AUC0-∞, t1/2 and Ke of escitalopram were 9.21±2.10 ng·ml-1 , 3.75±1.04 h, 514.6±152.3 ng·h·ml-1 ,540.5±162.3 ng·h·ml-1 , 34.06±7.71 h and 0.021±0.004 h-1,respectively. CONCLUSION:The determination of concentration of escitalopram in human plasma by HPLC-MS/MS method was repid, sensitive and reliable. It can be used for clinical pharmacokinetic study of escitalopram.

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