1.The role of jasmonic acid in stress resistance of plants: a review.
Lehuan ZHANG ; Changyu ZOU ; Tianxiang ZHU ; Meixia DU ; Xiuping ZOU ; Yongrui HE ; Shanchun CHEN ; Qin LONG
Chinese Journal of Biotechnology 2024;40(1):15-34
Jasmonic acid (JA), a plant endogenously synthesized lipid hormone, plays an important role in response to stress. This manuscript summarized the biosynthesis and metabolism of JA and its related regulatory mechanisms, as well as the signal transduction of JA. The mechanism and regulatory network of JA in plant response to biotic and abiotic stresses were systematically reviewed, with the latest advances highlighted. In addition, this review summarized the signal crosstalk between JA and other hormones in regulating plant resistance to various stresses. Finally, the problems to be solved in the study of plant stress resistance mediated by JA were discussed, and the application of new molecular biological technologies in regulating JA signaling to enhance crop resistance was prospected, with the aim to facilitate future research and application of plant stress resistance.
Signal Transduction
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Cyclopentanes
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Oxylipins
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Plant Growth Regulators
2.Correlation Analysis of ADPRT rs1136410 Polymorphism with the Occurrence of Non-small Cell Lung Cancer in Han Nationa- lity from Northern Jiangsu
Weiping HE ; Huaixue JI ; Shuqun HU ; Jingran CAI ; Huizhuo TANG ; Dongsheng PEI ; Xiuping DU ; Yan WANG
China Pharmacy 2019;30(16):2258-2262
OBJECTIVE: To investigate the correlation of ADPRT rs1136410 polymorphism with the occurrence of non-small cell lung cancer (NSCLC) in Han nationality from northern Jiangsu. METHODS: A total of 283 patients with primary NSCLC of Han nationality in Northern Jiangsu were selected from the Affiliated Hospital of Xuzhou Medical University during Nov. 2015-Dec. 2018 as NSCLC group. A total of 210 healthy subjects underwent physical examination were included in control group. PCR-RFLP was utilized to determine the genotypes at ADPRT rs1136410 locus. Logistic regression model was used to evaluate the effect of polymorphism and its interaction with smoking on the occurrence of NSCLC. RESULTS: There was no statistical significance in age and gender between 2 groups (P>0.05). The proportion of smoker in NSCLC group was significantly higher than control group (P<0.05). TT, TC and CC genotypes were detected at rs1136410 locus of ADPRT gene. The frequency of TT, TC and CC genotype were 41.9%,44.8% and 13.3%, and those of allele T and C were 64.3% and 35.7% in control group. The frequency of TT, TC and CC genotype were 21.6%, 50.2% and 28.2%, and those of allele T and C were 46.6% and 53.4% in NSCLC group, respectively. The frequencies of genotypes in 2 groups were in accordance with Hardy-Weinberg equilibrium (P>0.05), while there was significant difference in genotype and allele frequencies between 2 groups (P<0.05). Compared with TT genotype, the risk of NSCLC in individuals carrying TC and CC genotypes raised by 1.179, 3.122 folds [ORTC=2.179, 95%CI (1.435, 3.309), P<0.05; ORCC=4.122,95%CI(2.401,7.075),P<0.05]. Compared with individuals carrying TT genotype, the risk of NSCLC occurrence in non-smokers carrying TC and CC genotypes increased by 0.371, 1.328 fold [ORTC=1.371,95%CI (0.927,3.428),P<0.05; ORCC=2.328,95%CI (1.249,4.622),P<0.05]; and the risk of NSCLC occurrence in smokers carrying TC and CC genotypes increased by 0.928, 2.182 folds [ORTC=1.928,95%CI (1.257,2.957), P<0.05;ORCC=3.182,95%CI (1.760,5.754), P<0.05]. CONCLUSIONS: The rs1136410 locus mutant genotype of ADPRT gene is the risk factor of NSCLC in Han nationality from Northern Jiangsu, and smoking raises this risk of NSCLC occurrence in individuals with mutation genotypes of ADPRT rs1136410.
3.Correlation Analysis of XRCC1 rs25487 Polymorphism with the Occurrence of Lung Cancer
Yu MU ; Huaixue JI ; Shuqun HU ; Xing GAO ; Xiuping DU ; Weiping HE ; Rumeng WU ; Yan WANG
China Pharmacy 2018;29(12):1648-1651
OBJECTIVE:To investigate the correlation of XRCC1 rs25487 polymorphism with the occurrence of lung cancer. METHODS:A total of 208 patients with primary lung cancer of Han nationality in Northern Jiangsu selected from the Affiliated Hospital of Xuzhou Medical University during Sept. 2015-Jul. 2016 were included in lung cancer group. A total of 214 healthy volunteers of the hospital underwent physical examination were included in control group. PCR-RFLP was used to detect the genotypes at XRCC1 rs25487 locus,and Logistic regression model was used to evaluate the correlation of genotypes with the occurrence of lung cancer. RESULTS:There was no statistical significance in the distribution of age and gender between 2 groups (P>0.05). The proportion of smoker in lung cancer group was significantly higher than control group,with statistical significance(P<0.05). AA,AG and GG genotypes were detected at rs25487 locus of XRCC1 gene. The frequency of AA,AG and GG genotype were 43.5%,41.1%and 15.4% in control group and 28.8%,48.6% and 22.6% in lung cancer group,respectively. The frequencies of genotypes in 2 groups were in line with Hardy-Weinberg equilibrium(P>0.05),but there was statistical significance in genotype distribution between 2 groups(P<0.05). Compared with AA genotype,the risk of lung cancer in individuals carrying AG genotype increased by 2.265 fold [OR=2.265,95%CI(1.299,3.950),P=0.040;after corrected with gender,age and smoking history OR=2.309,95%CI(1.274, 4.185),P=0.006],with statistical significance. The risk of lung cancer in individuals carrying GG genotype increased by 1.310 fold [OR=1.310,95%CI(0.771,2.228),P=0.318;after corrected OR=1.429,95%CI(0.811,2.518),P=0.217],without statistical significance. CONCLUSIONS:rs25487 locus mutant heterozy-gosity of XRCC1 gene is risk factor of lung cancer in Han nationality from Northern Jiangsu,and smoking can increase the risk of lung cancer.
4.Analysis of Epidermal Growth Factor Receptor Gene Mutation in 274 Cases of Non-Small Cell Lung Cancer
Yan WANG ; Rumeng WU ; Xiaobing XIA ; Xiuping DU ; Huaixue JI
China Pharmacy 2018;29(20):2817-2821
OBJECTIVE:To study the gene mutation status of epidermal growth factor receptor(EGFR)in non-small cell lung cancer(NSCLC)patients and its relationship with clinical indexes,and to provide reference for individualized administration of EGFR-TKI in NSCLC patients. METHODS:Totally of 274 NSCLC patients from the northern of Jiangsu area were selected from our hospital during Jan. 2015-Dec. 2017. Mutation status of EGFR gene in lung tissue was determined by amplification refractory mutation system (ARMS)-TaqMan PCR assay. The relationship of EGFR gene mutation with clinical indexes as gender,age, smoking status, staging, tumor differentiation and pathological type were analyzed retrospectively. Compared with related literatures,the regional differences of EGFR gene mutation were analyzed. RESULTS:Among 274 NSCLC patients,112 patients suffered from EGFR gene mutation with total mutation rate of 40.88%. There were 50,57,3,2 cases of exon 19,exon 21,exon 20 and exon 19+21 mutation,and the types of EGFR gene mutation were delE746-A750,L858R and insH773-V774H,etc. The mutation rates of EGFR gene exon 19,exon 21 in non-smoking,early,well-differentiated and adenocarcinoma patients were 52.50%,47.24%,46.36% and 45.00%,which were significantly higher than smoking (28.57%),advanced (27.03%), poor-differentiated(31.71%)and squamous cell carcinoma(27.66%)patients,with statistical significance(P<0.05). There was no statistical significance in mutation rates of EGFR gene exon 19 and exon 21 between male and female,≥65 year-old and <65 year-old patients (P>0.05). EGFR mutation rate of NSCLC subjects from the northern of Jiangsu area was significantly higher than Shanghai area(P<0.05);there was no statistical significance compared with Yunnan area(P>0.05)but mutation types were different. CONCLUSIONS:There is the highest EGFR gene mutation rate in its exon 21,lesser in exon 19,rare in exon 20 and exon 19+21 among NSCLC patients from the Northern of Jiangsu area. There are obvious regional differences. The mutation rate of EGFR gene mutation exon 19 and exon 21 are associated with smoking status,staging,tumor differentiation and pathological type of NSCLC patients. The non-smoking, early stage, well-differentiated and adenocarcinoma patients are more likely to benefit from EGFR-TKI targeted therapy.
5.A multi-center clinical research of diagnostic value of serum gastrin-17 combined with pepsinogen for gastric cancer
Chunping ZHU ; Jianye ZHAO ; Xiaojun SHEN ; Wei QIAN ; Yingcai MA ; Shuo ZHANG ; Jianming XU ; Xiuping WAN ; Yiqi DU ; Zhaoshen LI
Chinese Journal of Digestive Endoscopy 2017;34(1):19-23
Objective To evaluate the diagnostic value of gastrin?17( G?17) and pepsinogen( PG) for gastric cancer. Methods A multicenter cross?sectional study of patients with continuous stomach discomfort from four centers including Changhai Hospital Affiliated to Second Military Medical University, the First Hospital Affiliated to Anhui Medical University, Qinghai Provincial People′s Hospital and the First Hospital Affiliated to Zhejiang University of Chinese Medicine from May 2014 to September 2015 was conducted. Before gastroscopy, fasting serum gatrin?17 and pepsinogen were analyzed by enzyme?linked immunosorbent assay(ELISA). The efficacy of G?17 and PG were evaluated according to endoscopic and pathological results. Results Based on the results of the pathological diagnosis, 1 122 cases were enrolled and divided into chronic atrophic gastritis group ( 548 cases ) , chronic non?atrophic gastritis group ( 370 cases), and gastric cancer group(204 cases). Serum G?17 and PGⅡ levels significantly increased(P<0?05) and PGR significantly decreased( P<0?05) in gastric cancer group compared with other groups. There was no significant difference in PGⅠlevel among three groups. The cut?off value of G?17 to diagnose gastric cancer was 7 pmol/L. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of G?17 for gastric cancer were 59?31%, 70?59%, 68?54%, 30?95% and 88?65% respectively. The cut?off value of PG Ⅰ/PG Ⅱ( PGR ) to diagnose gastric cancer was 7. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of PGR for gastric cancer were 41?18%, 83?01%, 75?40%, 35?00% and 86?39% respectively. The cut?off value of PGⅡto diagnose gastric cancer was 10 μg/L. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of PGⅡfor gastric cancer were 73?53%, 53?05%, 56?77%, 25?82% and 90?02% respectively. If G?17>7 pmol/L and PGR<7 was regarded as the cut?off value of diagnosis of gastric cancer, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 25?00%, 91?29%, 79?23%, 38?93%and 84?56%respectively. If G?17>7 pmol/L and PGⅡ>10μg/L was regarded as the cut?off value, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 48?04%, 79?74%, 73?98%, 34?51% and 87?35% respectively. If PGR<7 and PGⅡ>10 μg/L was regarded as the cut?off value, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 33?82%, 84?86%, 75?58%, 33?17% and 85?23% respectively. Based on logistic regression analysis of the independent variables of high serum G?17 value(>7 pmol/L), low serum PGR value(<7) and high serum PGⅡvalue(>10 μg/L), their OR value were 2?592, 2?237 and 1?864 respectively, and high serum G?17 value showed the highest risk of gastric cancer. Conclusion High serum G?17 and PGⅡ, low PGR are indicators of gastric cancer. Combination of G?17 and PGR has the best diagnostic value for gastric cacer. Gastric cancer can be screened in large scale by combining G?17 and PGR in order to improve the early diagnostic rate of gastric cancer and reduce the mortality of gastric cancer in our country.
6.Effects of apelin-13 on myocardium metabolism in diabetic rats
Mengze DU ; Jinghui FENG ; Hang ZHAO ; Ying LU ; Xiuping WU
Chinese Journal of Geriatrics 2017;36(4):454-458
Objectives To investigate the effects of Apelin 13 on myocardial metabolism in diabetic rats.Methods A total of 40 male Wister rats were randomly divided into normal control group (NC,n=8) and experimental group (n =32).Diabetic rats model were induced by high-sugar and high-fat diet combined with low-dose intraperitoneal injection of streptozotocin (STZ).The wellestablished 28 diabetic model rats were randomly divided into diabetic model group (DM,n=14) and apelin-13 treated group (n=14).In the Apelin-13 group,diabetic rats were administered Apelin-13 [0.1 μmol/(kg · d)]by intraperitoneal injection for 10 weeks,while the control group and diabetic model group were given an equal volume of 0.9% NaCl.At the end of the 10th week,all rats were sacrificed after fasting glucose measurement.Levels of serum free fatty acids (FFA) and myocardial FFA were measured by ELISA.Expression of myocardial glucose transporter member 4 (GLUT4) were detected by immunohistochemistry.The mRNA expressions of myocardial PPARα,CD36 and CPT-1 were detected by real-time fluorescence quantitative PCR.Results Fasting blood glucose,serum FFA and myocardial FFA were significantly higher in DM group than in NC group (all P< 0.05).The level of plasma glucose and myocardial FFA were significant lower(P>0.05) in Apelin-13 treated group than in DM group;but serum FFA was not significantly lower(P<0.05).The mRNA expressions of PPARα,CD36,CPT-1 in cardiac myocyte were higher in DM group and Apelin-13 treated group than in control(P<0.05),and lower in Apelin-13 treated group than in DM group(P< 0.05).The expression of myocardial GLUT4 was significantly lower in DM group(1.138±0.316)and in Apelin-13-treated group (4.631 ± 1.832) than in NC group(9.132 ± 2.156),(F=65.507,P< 0.05),and higher in Apelin-13-treated group than in DM group(P<0.05).Conclusions Apelin-13 increases myocardial expression of GLUT4,improves utilization of FFA,and it can effectively reduce the expression of PPARα,CD36 and CPT-1.Therefore,it may play a vital role in the improvement of myocardial metabolism in diabetic rats.
7.Prokaryotic expression and purification of GST-NRP-1 fusion protein
Zhengxiang HAN ; Mengjin ZHANG ; Jie XU ; Hongmei WANG ; Xiuping DU ; Chong CHEN ; Kailin XU
Chinese Journal of Immunology 2015;(10):1370-1374
Objective:To construct GST-tagged human NRP-1 fusion protein expression vector and induce its expression in Escherichia coli ( E.coli) ,then carry on inclusion body refolding and purification so as to obtain GST-NRP-1 fusion protein.Methods:NRP-1 gene was amplified by RT-PCR and inserted into pCR-blunt vector.Then the reconstructed plasmid was inserted into prokaryotic expression vector pGEX-4T-1.The constructed pGEX-4T-1-NRP-1 expression vector was transformed into BL21 cells and induced by i-sopropyl-β-D-thiogalactoside ( IPTG).Bacterial bodies were disrupted by sonication.Then the soluble fraction of fusion proteins were verified by Western blot and purified by Glutathione Sepharose 4B after inclusion body refolding.Results: The NRP-1 gene fragment was amplified by RT-PCR and inserted into pCR-blunt vector.Fusion protein expression vector pGEX-4T-1-NRP-1 was constructed suc-cessfully.After transformation, GST-NRP-1 expression vector was detected in BL21 cells and obtained purifying protein after refolding.Conclusion:The plasmid GST-NRP-1 was constructed successfully and laid basis for subsequent studies.
8.Effects on proliferation and apoptosis of T-cell leukemia cells by silencing NRP-1
Hongmei WANG ; Zhenyuan XU ; Xiuping DU ; Taotao LI ; Zhengxiang HAN
Chinese Journal of Immunology 2015;(11):1447-1452
Objective:To investigate the effect on proliferation and apoptosis of T-cell leukemia cells by silencing NRP-1 ( Jurkat cells).Methods:The lentivirus plasmid which expresses NRP1 gene specific shRNA was constructed in our preliminary ex-perimental.We transfected the lentivirus plasmid to human T-cell Lymphoma cells.The proliferation of Jurkat cells different groups and effect on cell proliferation after chemotherapy drug EPI-treated were found by CCK-8 kit.The proliferation level and apoptosis rate of the cells were detected by flow cytometry and Annexin-V-FITC/PI method.Results:The proliferation level of NRP-1 /shRNA interference group was decreased significantly in 48 h,72 h,96 h,which was compared with the control groups.The apoptosis rate of the NRP-1/shRNA interference group was increased compared with control groups.The chemotherapy drug sensitivity of epirubicin ( EPI ) test results showed that EPI concentration was 0.025,0.05,0.1,0.2,0.4 μg/ml,the NRP-1/shRNA interference group of cell growth inhibition rate was increased,the corresponding control group difference had statistical significance(P<0.05).We choose the drug con-centration of the EPI IC50 for next experiments.NRP-1/shRNA interference group cell apoptosis rate increased significantly after induction,compared with the control groups difference was statistically significant ( P<0.05 ).Compared with control group, the expression level of Bcl-2 protein was decreased and the expression level of bax protein was increased significantly after EPI induction.The percentage of cells at G0/G1 phase increased significantly,while those at S phase decreased significantly.Conclusion:Plasmid shRNA-NRP1 inhibited the expression of NRP1 in Jurkat cells and decreased the proliferation level of Jurkat cells and promote their apoptosis and enhance their drug sensitivity;the molecular mechanism may relate to down-regulation of Bcl-2 and up-regulation of Bax.and arrested the cell cycle at G0/G1 phase.
9.Preliminary study of the differences of related gene expression in cancer stem cells of human breast cancer cell lines MCF-7 and MDA-MB-231
Zhengxiang HAN ; Jie ZHANG ; Jie XU ; Xiuping DU
Cancer Research and Clinic 2015;27(6):402-405
Objective To enrich breast cancer stem cells of breast cancer cell lines MCF-7 and MDA-MB-231 through culturing mammospheres,and to detect the expression differences of gene of breast cancer stem cells makers.Methods The MCF-7 and MDA-MB-231 cell lines were cultured by serum and serum-free medium.The proportion of CD44+/CD24-and CD133+ cancer stem cells was measured in cells derived from mammosphere cells or monolayer culture cells by flow cytometry,and the expression of CD44,CD24,CD133,ALDH3A1,ABCG2 and CXCR4 mRNA were detected by RT-PCR.Results Flow cytometry analysis indicated that the CD44+/CD24-low proportion of the MCF-7 mammosphere cells was higher than its adherent culture cells (P < 0.05),and CD133+ proportion had no difference between them (P > 0.05).However,the CD44+/CD24-/low proportion of the MDA-MB-231 mammosphere cells was lower than its adherent culture cells (P < 0.05),while CD133+ proportion was significantly higher than its adherent cultured cells (P < 0.05).RT-PCR analysis suggested that the expression of CD44 and ABCG2 increased obviously in MCF-7 microspheres (P< 0.05),and the expression of CD24,CD133,ALDH3A1 and CXCR4 had no significant difference between the mammosphere cells and adherent culture cells (P > 0.05).The CD24,CD133,ABCG2,ALDH3A1 and CXCR4 increased obviously in MDA-MB-231 microspheres.On the contrary,the CD44 decreased (P < 0.05).The expression of CD44 and CD24,CD133 and ALDH3A1 had significant differences in the microspheres of MCF-7 and MDA-MB-231 cells (P < 0.05).Conclusion The related cancer stem cells gene expression is different in the microspheres of human breast cancer cell line,which indicates that the different subtypes of breast cancer may be derived from different origins.
10.Analysis of clinical effect of recombinant human granulocyte-macrophage colony-stimulating factor combined with recombinant bovine basic fibroblast growth factor in the treatment of oral ulcer caused by chemotherapy
Xuan SUN ; Xiuping DU ; Zhengxiang HAN
Cancer Research and Clinic 2015;27(4):263-265
Objective To investigate the effect of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) combined with recombinant bovine basic fibroblast growth factor (rb-bFGF) in the treatment of oral ulcer caused by chemotherapy.Methods 108 patients of oral ulcer caused by chemotherapy were randomly divided into two groups.54 cases in the control group were treated with cydiodine buccal tablets at first,then received the aerosol treatment which was prepared by mixing gentamicin,dexamethasone,2 % lidocaine and physiologic saline,three times per day.54 cases in the treatment group firstly received the gargle which was prepared by mixing rhGM-CSF,dexamethasone and physiologic saline,then were treated with rb-bFGF by spraying on the oral ulcer surface,three times per day.Results The effective rate of the treatment group was 96.30 % (52/54),which was significantly higher than that of the control group [64.81% (35/54)],there was a significant difference between the two groups (x2 =17.08,P < 0.05).Conclusion The effect of rhGM-CSF combined with rb-bFGF in the treatment of oral ulcer caused by chemotherapy is very significant.

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