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OsRhoGDI2 was isolated as a putative partner of Rho protein family member OsRacD from rice panicles by yeast two-hybrid, but its function remains unknown. In order to identify the function of OsRhoGDI2, OsRhoGDI2 knockout mutants were created by CRISPR/Cas9 technology. The results showed that two different homozygous mutants were obtained in T0 generation, and eight kinds homozygous mutants were identified in T1 generation. Sequence analysis revealed that the base substitution or base deletion occurred near the editing targets of the gene in knockout rice, and it could be expected that the truncated OsRhoGDI2 proteins lacking the RhoGDI conserved domain would be generated. Phenotype analysis showed that the OsRhoGDI2 knockout rice plants were significantly lower than the control plants. Statistical analysis confirmed that the significant decrease of plant height was due to the shortening of the second and third internodes, suggesting that OsRhoGDI2 gene may be related with rice height control.
Subject(s)
CRISPR-Cas Systems , Genes, Plant , Genetics , Oryza , Genetics , Plants, Genetically Modified , rho Guanine Nucleotide Dissociation Inhibitor beta , GeneticsABSTRACT
Objective:To compare the prognosis of fluoroucil combined with cisplatin and paclitaxel combined with cisplatin regimens with concurrent radiotherapy in treatment of esophageal squamous cell carcinoma.Methods:A total of 120 patients with esophageal squamous cell carcinoma who were admitted to Anyang Tumor Hospital of Henan Province from December 2012 to November 2018 were randomly divided into group A and group B by using a random number generator. Group A was given cisplatin combined with 5-fluorouracil, and group B was given cisplatin combined with paclitaxel. Both groups had the same radiotherapy regimen, and both used intensity-modulated radiation therapy (IMRT). Completions of 50 Gy radiotherapy and at least one cycle of chemotherapy were considered to be in line with the plan. Survival data was analyzed in the term of intention-to-treat (ITT) and per-protocol (PP) set.Results:Of the 120 patients, 114 patients were treated and the adverse reactions could be evaluated, including 55 cases in group A and 59 cases in group B. The incidence of grade Ⅲ-Ⅳ leukopenia in group B was higher than that in group A [49.2% (29/59) vs. 25.5% (14/55)], and the difference was statistically significant ( χ2 = 6.805, P = 0.012), and there were no statistical differences in the other adverse reactions between the two groups (All P > 0.05). A total of 113 cases can be analyzed for survival. According to ITT analysis, the median progression-free survival (PFS) time in group A and group B was 28.0 months (95% CI 15.5-34.5 months) and 27.0 months (95% CI 17.0-41.0 months), the median overall survival (OS) time was 28.0 months (95% CI 15.8-34.2 months) and not reached, the differences were not statistically significant (both P > 0.05). According to PP analysis, the median PFS time in group A and group B was 28.0 months (95% CI 15.8-34.2 months) and 29.0 months (95% CI 14.9-45.1 months), the median OS time in group A and group B was 28.0 months (95% CI 3.7-52.3 months) and not reached, the differences were not statistically significant (both P > 0.05). Conclusions:The fluorouracil combined with cisplatin regimen and paclitaxel combined with cisplatin regimen with concurrent radiotherapy have similar PFS and OS time in treatment of esophageal squamous cell carcinoma, the adverse reactions are different, but they are all tolerable. In individualized clinical practice, the toxicities and costs of the two regimens can be comprehensively considered.
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Objective To analyze copy number variance (CNV) in whole genome by using gene chip technology, and to screen the radiosensitivity associated genes on esophageal squamous cell carcinoma (ESCC). Methods The patients with ESCC who received radiotherapy alone in Anyang Tumor Hospital from December 2013 to August 2016 were selected, and biopsy paraffin samples were preserved in the center of pathology. The patients were divided into radiosensitivity group (group S) and radio-resistance group (group R). DNA was extracted from these paraffin samples in both groups. Whole human genome CNV was detected by using genechip from OncoScan Array platform designed by Affymetrix company, and the differences of gene segments were screened in the two groups. Results Nineteen samples of ESCC patients were collected to extract DNA in this study. To balance pair analysis in the two groups, 10 samples were selected from the qualified patients, including 5 cases in group S and 5 cases in group R respectively. There were no statistical differences in gender, age, lesion site, lesion length, radiation dose of the two groups (all P> 0.05). Loss of heterozygosity (LOH) was the main type of CNV. The analysis results showed that LOH in q24.32-q24.33 of chromosome 10 and LOH in q21.2-q21.31 of chromosome 18 had high frequencies (100 ％) in group R, however, none were detected in group S. LOH in q27-q28.1 of chromosome 4 had a high frequency (80％) in group S , however, none were detected in group R. Conclusion LOH in 10q/18q is related to radio-resistance in ESCC, and LOH in 4p is associated with radiosensitivity in ESCC.
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Objective To investigate the effects of radiotherapy of patients with post-operative locoregional recurrence of esophageal carcinoma.Methods Retrospectively analyzed the clinical data of 60 patients with esophageal carcinoma who developed post-operative loco-regional recurrence.There were 8 patients with anastomosis relapse,21 with anastomosis and mediastinal lymph nodes metastasis,5 with abdominal lymph node metastasis,9 with supra clavicular lymph node metastasis,11 with mediastinal lymph node metastasis and 6 with mediastinal and supra-clavicular lynph node metastasis.Three-dimensional conformal radiation therapy was used to treat all the patients,with a total radiation dose of 50-70 Gy.Short-term effect,survival and adverse reactions of acute radiation were observed after radiotherapy.Results The effective rate of short term was 78.3％.The 1st,2nd and 3th year overall survival rates were 100％,50.0％,30.0％,respectively,and the middle survival phase was 15.8 months.Acute radiation side effects include:16 cases of acute radiation esophagitis grade Ⅰ,Ⅱ (26.7％,16/60),4 cases of Ⅲ grade (6.7％,4/60);11 cases of acute radiation pneumonitis (18.3％,11/60);3 cases of bone marrow suppression Ⅰ,Ⅱ grade (5％,3/60) and acute cardiac injury in 1 case (1.7％,1/60).Conclusion Radiotherapy may improve the survival rate and the reduce the therapy phase of esophageal carcinoma patients with postoperative recurrence which is a good method for patients.