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.

Objective To explore the reliability and validity of driver coping questionnaire (DCQ) Chinese version. Methods Three hundred and forty drivers were investigated at random by DCQ and positive affect nagative affect scale (PANAS). Results Four factors were extracted, namely active appraisal and coping, confrontive coping, emotion-focused coping and avoidance coping. The correlation coefficients between the questionnaire and the four factors were 0.317 ～ 0. 813 (P ＜ 0.01 ), and the correlation coefficients among the four factors were 0.057 ～0.294. Positive appraisal and coping were correlated with PA positively( r=0. 171, P＜0. 01 ) ,and with NA negatively( r= -0. 349, P＜0.01 ),and the other three coping styles were positively correlated with NA( r=0. 242 ～0.262, P＜0.01 ). The split-half reliability was 0.734,the Cronbach α coefficients of the total questionnaire and four factors were 0. 807,0. 890,0. 621,0. 571and 0. 687 respectively. Conclusion DCQ Chinese version has good reliability and validity, and is an effective instrument to test drivers' coping behavior.

Objective To observe the effect of tanshinoneⅡA (TⅡA) on arrhythmia and levels of ischemia modified albumin (IMA) and heart type-fatty acid binding protein (H-FABP) in rats with myocardial ischemia-reperfusion injury.Methods Forty Wistar rats were equally randomized into 5 groups:pseudo-operation group (normal saline),model group (normal saline),and high-,middle-and low-dose TⅡA groups (in the dose of 20,10 and 5 mg?kg-1?d-1 respectively).The rats were treated for 7 days,and received operation 2 hours after the last medication.Rats in pseudo-operation group received threading without ligation,while animal model of myocardial ischemia-reperfusion injury was established by ligating the left-anterior coronary artery for 15 minutes and then loosening for 30 minutes.During the operation,electrocardiogram was traced continuously to observe the arrhythmia.HE staining was used to observe the pathological changes of myocardium.Levels of IMA and H-FABP in myocardium were determined by enzyme-linked immunosorbent assay (ELISA).Results Ischemic changes of the myocardium and arrhythmia occurred,myocardial IMA level increased and H-FABP level decreased in the model rats.TanshinoneⅡA obviously attenuated arrhythmia caused by ischemia-reperfusion injury (P

BACKGROUND: siRNA transfection is a key step in RNA interference. The methods of cardiomyocytes transfection were most use of plasmids as vector to transfect long-chain shRNA. However, the processes were complex. It was a simple efficient and low-toxic method that liposomes-mediatad chernosynthesis siRNA transfection. It was useful for expanding RNA interference application.OBJECTIVE: To choice the optimal concentration of Iiposomes-mediated chemosynthesis siRNA transfection, and to discover a simple efficient RNA interference application.METHODS: CY3-Negative siRNA was mediated by lipid-besed agent siPORT~(TM) NeoFX~(TM) to transfect cardiomyocytes. A blank control group was set. After 24 hours, the transfection efficiency and apoptotic rate were evaluated by fluorescent microscope and flow cytometer to select an optimal concentration. Based the best concentration, siRNA PHB was transfected to cerdiomyocytes. 48 hours later, the expression of PHB was tested.RESULTS AND CONCLUSION: With the increased concentration of CY3-Negative siRNA, the number of cells emitted red fluorescence grew under fluorescence microscope, and the transfection efficiency was also increased (P＜0.05). The best concentration was 30 nmol/L (P＜0.05). There was no significant difference in apoptotic rate between transfected groups and the control group (P ＞ 0.05). The PHB expression of cardiomyocytes transfected siRNA PHB was dropped by 74.11% on average (P＜0.05). The results indicated that lipid based agent siPORT~(TM) NeoFX~(TM) was suitable to transfect chemosynthesis siRNA to cardiomyocytes, and the best transfection concentration of siRNA was 30 nmol/L.

Objective To investigate the anti-apoptotic mechanism of tanshinone ⅡA and the function of prohibitin(PHB)onmyocardial cells apoptosis induced by hydrogen peroxide(H2O2).Methods Myocardial cells were primary culturedneonate rat were cultured in medium with 200 μmol/L H2O2,and the medium was supplemented with tanshinone ⅡA(1 × 10-4 mol/L)in advance for 24 h.PHB in myocardial cells was knocked down by RNA interference,and theexpression level of PHB was determined by Western blotting analysis.Flow cytometric analysis was used to detectapoptosis rate,intracellular calcium concentration([Ca2+]i),and mitochondrial membrane potential(MMP).ResultsH2O2-mediated cell apoptosis resulted in activation of PHB,increasing of[Ca2+]i,and decreasing of MMP.TanshinoneⅡA profoundly inhibited myocardial cell apoptosis induced by H2O2,decreased[Ca2+]i,and increased MMP.Specificsilence of PHB by siRNA down-regulated the expression level of PHB,increased apoptosis rate and[Ca2+]i,and decreasedMMP.Conclusion The results demonstrate that tanshinone ⅡA could attenuate apoptosis induced by H2O2,and theactivation of PHB induced by H2O2 is the major regulatory pathway of cyto-protective gene expression against oxidativestress.

10-15 years were 7 cases(6.3%),they were randomly divided into treatment group and contol group.The treatment group with 60 patients were given INF-? and polyresistin,age 3 years,given IFN-? 200 000 U per day;and polyresistin 5 mg per time,3 times every day,the course of therapy were 5 days.The control group was given traditional Chinese medicine at the same time.Results After therapy,in treatment group,53 cases(88.3%) of children′s cooling time were lower than 5 days,47 cases(78.3%) of children′s rash subsided time were lower than 6 days,48 cases(75%) of children′s Catarrhal symptoms disappeared time were lower than 5 days,and the rate of complications′ occurrence were 15 cases(18.3%).In control group,28 cases(53.8%) of children′s cooling time were lower than 5 days,14 cases(26.9%) of children′s rash subsided time were lower than 6 days,28 cases(53.8%) of children′s Catarrhal symptoms disappeared time were lower than 5 days,and the rate of complications′ occurrence were 37 cases(71.5%),there were significant differences between both groups(Pa

Corneal endothelial cell(CEC)is the most critical part for the cornea, of which activity can influence the postoperative vision.It is very important for the clinical cornea preservation considering the function and its self-purification of donor cornea.There are a variety of classical methods, which can significantly prolong the saving time of donor cornea with its good quality of CEC.We reviewed the published papers about present preservation methods of cornea, which can give us many suggestions for the clinical cornea preservation.

Psoriasis is a chronic,refractory,inflammatory skin disease that occurs in young adults.The traditional animal model cannot simulate the skin characteristics of patients with psoriasis effectively, so it is difficult to be used for in-depth study of psoriasis mechanism. Immortalized human epidermal cells (HaCaT)is a non-tumor,immortalized human epidermal cell which is widely used in the study of dermatosis.HaCaT cells are the best choice for the study of psoriasis mechanism because their immu-nological characteristics and reproductive ability are coincide with the pathological features of psoriasis. This article reviews the specific methods such as establishment of cell method, cytokine and chemo-kine analysisin the pathogenesis study of psoriasis based on HaCaT cells, hoping to provide some thoughts for drug′s pharmacological activity research.

Objective To reproduce and validate the hexokinase reference method for glucose detection and compare other 5 routine glucose kits with this reference method. Methods The CDC hexokinase reference method for glucose detection was established and the performance was validated through testing a standard reference material (SRM) and participating in the IFCC ring-trial for reference laboratories for glucose evaluation. The CLSI EP 9-A2 protocol was used to compare the 5 routine glucose kits with the hexokinase reference method. Forty serum samples were analyzed by 5 routine kits and the hexokinase reference method. Results When SRM 965a was determined by the reference method,the bias of level 2 and level 3 were 0. 93%, -0. 23% respectively. The results for IFCC ring trial were within the accepted range. For the 5 routine kits, the confidence intervals of the predictive bias at the medical decision point Xc (Xc = 6. 11 mmol/L) were all within the range of defined acceptable error (10%) and the range of biological variation bias (6.9%). Conclusions The hexokinase reference method for serum glucose was reproduced in our lab. The serum glucose results measured with 5 routine kits were different from results detected with the reference method, but the bias was acceptable,and it will not affect the detection results.