To explore the influence of low light on the synthesis of carotenoids, chlorophyll and the adaptability of transgenic plants with tomato Solanum lycopersicon L. GGPS2 gene, we constructed a vector containing a GGPS2 gene with green fluorescent protein (GFP) as report gene under the control of a cauliflower mosaic virus 35S promoter and introduced it into tobacco Nicotiana tabacum L. cv. Wisconsin 38 by Agrobacterium tumefaciens-mediated transformation. PCR analysis of the DNA from kanamycin resistant tobacco indicated that the transgenic tobacco containing the nptII gene, SlaGGPS2 gene and without contamination of Agrobacterium. We also detected the root tip of kanamycin resistant tobacco showing characteristic fluorescence. The contents of carotenoid, chlorophyll and photosynthesis of transgenic tobacco increased in comparison with wild tobacco after low light treatment. In addition, leaf mass per unit area, total dry weight, ratio of root to shoot in transgenic tobacco were all higher than that of the wild tobacco, which proved that the transgenic tobacco could increase the accumulation of biomass and promote it transport to root. The transgenic tobacco with SlaGGPS2 gene can increase the contents of carotenoid, chlorophyll, enhance the photosynthetic rate, promote the biomass accumulation and its distribution to root. Hence, the transgenic tobacco with SlaGGPS2 gene had increased low light tolerance and the SlaGGPS2 gene maybe can be used in other crops.
Agrobacterium tumefaciens ; Carotenoids ; analysis ; Chlorophyll ; analysis ; Gene Expression Regulation, Plant ; Genetic Vectors ; Light ; Lycopersicon esculentum ; genetics ; Photosynthesis ; Plants, Genetically Modified ; metabolism ; radiation effects ; Tobacco ; metabolism ; radiation effects

Objective To investigate the influence of ocular residual astigmatism (ORA) on the correction of astigmatism by FS-LASIK with vector analysis.Methods The records of 182 patients who had accept FS-LASIK between January,2016 and April,2016 were retrosepectively reviewed.The patients whose ORA ≥ refractive cylinder were assigned to high ocular residual astigmatism group (HORA group),ORA ＜ refractive cylinder were assigned to low ocular residual astigmatism group (LORA group).All of the patients were followed 6 months or more.The visual acuity,error ratio and correction ratio were compared between HORA group and LORA group.Results The preoperative ORA of all patients was (0.61 ± 0.27) D,in which ＞ 0.75 D were 58 cases (31.9％),and the HORA group was more than the LORA group (P ＜ 0.05).At postoperative 6 months,there was no statistically significant difference in vision acuity between the HORA group (1.06 ± 0.15) and LORA group (1.08 ± 0.15) (t =0.97,P =0.35).There was statistically significant difference in the error ratio between the HORA group (58.11 ± 63.23) ％ and LORA group (26.12 ± 35.37) ％ (t =3.43,P ＜ 0.05).There was statistically significant difference in the correction ratio between the HORA group (146.45 ± 86.63) ％ and LORA group (122.56 ± 36.31) ％ (t =2.81,P ＜ 0.05).Conclusion The error ratio and correction ratio of astigmatic correction by FSLASIK is significantly higher in eyes with high ORA than in eyes with low ORA.Vector analysis should been carried out before the FS-LASIK.

Objective:Use the droplet digital PCR (ddRCR) technology to establish, optimize and evaluate the method of EGFR-T790M mutation detection.Methods:The relevant probes and primers were designed for EGFR-T790M mutations. The ddPCR reaction system was established, the optimal annealing temperature was set and the basic performance of the method was tested. On this basis, from January 2019 to October 2019, 72 cell-free DNA (cfDNA) samples from NSCLC patients were collected from Shandong Cancer Hospital Affiliated to Shandong First Medical University, and clinically verified. The consistency of the gene mutation detections with Bole ddPCR products was analyzed using Kappa test.Results:The ddPCR reaction system was established and optimized. Linear evaluation showed the R2 value was greater than 0.99. Using ddPCR, the blank detection limit was determined to be the numbers of mutant droplets≥2, with excellent specificity. For the sensitivity analysis, the lower limit of mutation detection was determined to be at least 0.05%. In the repeatability and inter-assay precision tests, the results had a coefficient of variation( CV)<20%. The relative deviation of the results was within the range of±10% for the accuracy analysis. Using the established T790M mutation detection method, 72 samples from the NSCLC patients were tested for genetic mutation in cfDNA, and the overall agreement with the Bole ddPCR products was 91.67% (66/72, Kappa=0.749; P<0.001). Conclusion:Using ddPCR, the method of EGFR-T790M mutation detection for NSCLC was successfully established.

Objective To summarize the clinical efficacy of renal transplantation from donors of donation after brain death (DBD) complicated with acute kidney injury (AKI). Methods Fifty-nine DBD donors successfully undergoing renal transplantation were recruited in this investigation. According to the Scr level upon admission of intensive care unit (ICU), DBD donors were divided into the AKI group (n=14) and control group (n=45). A total of 101 recipients were assigned into the AKI group (n=23) and control group (n=78) correspondingly. The organ donation conditions of 59 donors were summarized. Main parameters of the donors before organ procurement were statistically compared between two groups. Postoperative kidney function, hospitalization condition and clinical outcomes of the recipients were statistically compared between two groups. Results Among 59 donors, 14 cases (24%) suffered from AKI. Two donors received continuous renal replacement therapy during organ maintenance. Compared with the donors in the control group, the APACHE Ⅱ score of the donors was significantly higher (P<0.05), the incidence of central diabetes insipidus was considerably higher (P<0.01), the Scr levels at admission of ICU and before organ procurement were significantly higher (both P<0.01) and the amount of urine at 24 h before organ procurement was dramatically less in the AKI group (P<0.01).Compared with the recipients in the control group, the Scr levels at postoperative 2 and 3 d were significantly higher (both P<0.05), the length of hospital stay was considerably longer (P<0.01) and the hospitalization expanse was significantly higher in the AKI group (P<0.05). No statistical significance was observed in the postoperative delayed recovery of renal graft function, incidence of acute rejection, infection and rehabilitation dialysis in the recipients between two groups (all P>0.05). At 3 months after transplantation, the recipients in two groups were discharged and the graft survival rate was 100%. Conclusions For renal transplantation from DBD donors complicated with AKI, active measures should be taken to maintain the organ and relieve the AKI, which yields similar clinical efficacy to renal transplantation from non-AKI donors and widens the origin of kidney graft.