In plant genetic engineering, selectable marker is needed to distinguish transformant. As the commercialization of transgenic plants, people are more and more paying close attention to their safety, among which mainly refers to the safety of selectable marker. In order to increase the safety of transgenic plants, biologists began to search for biosafe selectable marker.Current research progress of taking key enzyme in metabolic pathways were overviewed,which include glycometabolism, amino acid metabolism, hormone metabolism ,nucleotide metabolism and protein metabolism et cetera, as selectable marker in transgenic plants.

The rodipicolinate synthase gene was cloned from Nicotiana tabacum L.cv,which encodes the key enzyme in the synthesis of lysine,and then a vitro molecular reform reliefing feedback suppression was made. Some resistant seedlings were acquired using the mutated gene as selectable marker and analog of lysine as selectable reagent,and the detection results were positive by means of PCR and Real-Time PCR,however the phenotype is somewhat abnormal.

Heart ; drug effects ; Heart Failure ; drug therapy ; Humans ; Hydrogen Bonding ; Nitrogen Oxides ; pharmacology ; therapeutic use

Malignant biliary tract obstruction is a common disease.Nowadays,endoscopic biliary drainage (EBD) and percutaneous transhepatic biliary drainage (PTBD) have been used to treat such disease.EBD is a first-line intervention for lower segment of biliary duct obstruction caused by pancreatic cancer or other tumors.PTBD is a first-line treatment for gallbladder cancer or other tumors which lead to the hilar and intrahepatic bile duct obstruction and abnormal digestive anatomy.In a word,the proper selection of the drainage should be based on the patients' condition with minimal trauma and maximum therapeutic effect.

Objective To develop a one-step PCR assay for rapid discrimination of six Brucella species and some intraspecific biovars.Methods Using 6 pairs of primers in one-step PCR to differentiate six classical Brucella species and some biovar in ordinary PCR instrument.The tested strains including 27 reference strains of six Brucella species and 239 Brucella strains were estimated by the PCR assay and biological identification methods.Results The six Brucella species could be precisely differentiated by the onestep PCR assay from the tested strains.Five biovars and vaccine strain of B.suis species could be determined,and biovars 1,3,4 and biovars 2,5,6,7,9 of B.abortus species could be identified at the level of their biovar,moreover,biovars 1,2 and 3,and vaccine strain Rev 1 of B.melitensis species were also discriminated at the biovar and strain level.The accurate rates of the biological identification method and the PCR assay were 98.33％ and 100％ respectively.Conclusion One-step PCR assay was a rapid,specific,and low cost method for identification of Brucella species and discriminating biovars in ordinary PCR instrument.

AIM:To put forward three-level design(Box-Behnken design)for fitting response surfaces to optimize the extraction technology of total flavonoids from Hypericum ascyron L.METHODS:Four factors,including temperature,reflux time,concentration of ethanol,ratio of solvent to raw material were used to examine the yield of total flavonoids.Prediction was carried out through comparing the observed and predicted values.RESULTS:The results suggested that ethanol concentration and extraction temperature were two statistically significant factors.The optimum conditions of extraction process consisted of the ratio(mL:g)of solvent to material(13.3:1),ethanol concentration(53.2%),extraction temperature(78.7℃)and extraction time(2.3h).Regression coefficient of binomial fitting complex model was as high as 0.984 6.Bias between observed and predicted values was-4.01%.CONCLUSION:Box-Behnken design is success in optimizing the extaction in close agreement with the predicted values of the mathematic model.

Objective:To analyze the biomechanical stress distribution on the sagittal fractured of the mandibular condyle(SFMC) fixed by double plates in 3 different ways.Methods:The fixation finite element models with double straight plates(SS),one straight and one L-form plates(SL),and double L-form plates(LL)were established.The biomechanical stresses on condyle were calculated with finite element analysis.Results:In the model of SS fixation the maximum equivalent stress (MES),maximum total displacement (MTD)and maximum total corner(MTC)of condylar stump were 525.475 MPa,0.902 2 mm and 0.260 1°;MES,MTD,and MTC on fractured free-end were 4.425 MPa,0.882 7 mm and 0.019 9°,respectively.In the model of SL fixation MES,MTD and MTC on condylar stump were 1 135 MPa,0.9367 mm and 0.126 1°;MES,MTD and MTC on fractured free-end were 2.656 MPa,0.887 1 mm and 0.016 9°,respectively.In the model of LL fixation MES,MTD and MTC on condylar stump were 2 208 MPa,0.923 8 mm and 0.172 2°;MES,MTD and MTC on fractured free-end were 14.66 8 MPa,0.876 6 mm and 0.018 1°,respectively.Conclusion:Double L plates fixation is a proper way for SFMC.

Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality ofAR. This study supplied a new method for the comparison of extract of herbal drugs.

Objective To observe the effects of different quantities of cake-separated moxibustion on blood cells and immunoglobulin in immunosuppressive rabbits; To study the possible mechanism of quantity-effect difference. Methods Immunosuppressive rabbit model was established by intraperitoneal injection of cyclophosphamide. Rabbits were randomly divided into blank group, model group, cake-separated moxibustion 3 Zhuang group, 5 Zhuang group, 7 Zhuang group.Liuwei Dihuang Decoction was used to make herbal-cake, and was put on Shenque (RN8), Guanyuan (RN4), and other acupoints, once for every other day, 10 times in total. Venous blood was taken on the next day after finishing treatment, and the contents of blood cells series and IgG, IgM, complement C3 and complement C4 were detected.Results Compared with blank group, the levels of WBC, NEU, RBC, HGB, HCT, PLT, IgG, IgM, and complement C3 of model group significantly decreased, while the levels of LYM and MONO significantly increased (P<0.01). Compared with model group, the increase of HCT and IgM in the cake-separated moxibustion 3 Zhuang group was not significant, the levels of WBC, NEU, RBC, HGB, HCT, PLT, IgG, IgM, and complement C3 in all treatment groups significantly increased, and LYM and MONO significantly decreased (P<0.01). Compared with cake-separated moxibustion 3 Zhuang group, the levels of WBC, IgG, and complement C3 of cake-separated moxibustion 5 Zhuang group significantly increased, and the levels of WBC, HGB and complement C3 of cake-separated moxibustion 7 Zhuang group significantly increased (P<0.01).Conclusion Cake-separated moxibustion can improve the immune function under immunosuppressive state induced by cyclophosphamide, and different quantities have differences in efficacy.

Embryo Transfer ; Endometrium ; Female ; Fertilization in Vitro ; Humans ; Integrative Medicine