Diterpenes, an important class of natural compounds, are widely distributed in nature. As the valuable diterpenoids continue to be found, diterpene synthase in the course of diterpene synthesis get as much attention as possible. The multiformity of end-product-diterpenoids were also due to the diversity of diterpene synthase. This paper focuses on the advances in recent biosynthesis pathway of diterpene and types, cloning, catalytic mechanism, synthetic biology application.

The transcription factor of ethylene responsive factor binding protein (ERF) is belonged to AP2/ERF superfamily, which is known to be unique in plants. AP2/ERF proteins have important functions in the transcriptional regulation of a variety of biological processes related to growth and development, as well as various responses to environmental stimuli. An ERF gene from Salvia miltiorrhiza is cloned and divided into ERF gene family group VII of Arabidopsis and Rice. It contains a MCGGAI (I/L) motif referred to as CMVII-1 and a single intron in the 5'-flanking region of the AP2/ERF domain. Sequence analysis reveals that the region of second extron has abundant polymorphism sites. There are 21 single nucleotide polymorphism sites (SNPs) in the 264 bp region, among them, 14 SNPs are synonymous substitutions and 7 SNPs are non-synonymous substitutions. Though analysis of 181 samples from Shandong, Shaanxi and Sichuan Provinces, it reveals that each production area has its own special genotypes, 5 SNPs show significant difference. Cluster based on UPGMA method reveals that different populations from specific province have clustered together. It shows that SmERF gene will be a candidate molecular marker for the identification of Salvia miltiorrhiza from different areas.

Objective A new breed of licorice seeds(Wuxin No.1) was selected from wild licorice,Glycyrrhiza uralensis.Methods Its germination rate,content of soluble protein,and peroxidase(POD) activities were investigated and compared with the wild licorice seeds(control group).The leaves of Wuxin No.1 were collected and its genetic polymorphism was analyzed by inter-simple sequence repeat technique(ISSR).Results The results suggested that the seed vitality in Wuxin No.1 was higher than that in the control group.The change of soluble protein and POD activity also demonstrated that the seeds in Wuxin No.1 have the higher vigor.ISSR Analysis showed that among 22 random primers used in this experiment,six primers generated different DNA band types,which meant that there was genetic polymorphism in Wuxin No.1.Conclusion All these changes indicate that Wuxin No.1 is a prospective domestication species of licorice and may be cultivated widely in the future.

Objective To construct eukaryotic expression plasmid of human B7-H1 extracellular region-hIgG1Fc-pCI-neo eukaryotic expression vector,and express the functional fusion protein in mammalian CHO cell.Methods Full-length human B7-H1 encoding sequence was amplified from human activated T cell cDNA library by PCR,fused with hIgG1Fc,then transformed into pCI-neo expression vector and verified by sequencing.The validated recombinant was transfected into mammalian CHO cell by lipofectamine reagent.The supernatant of the cultured cell was collected and analyzed by the sandwhich ELISA to detect if there was the fusion protein,and the fusion protein was purified by HiTrap recombination protein Protein A affinity chromatography.The concentrated supernatant or purified fusion protein were used for Western blotting after SDS-PAGE to identify the molecular weight and immune activity of the fusion protein of B7-H1.Results The extracellular region of hB7-H1 about 727 bp was cloned from human T cell cDNA library and was inserted with hIgG1Fc into the eukaryotic expression vector pCI-neo.After the transfection of recombinant into mammalian CHO cell by lipofectamine reagent,the expression of B7-H1 fusion protein was detected in the cultured CHO cell supernatant by the sandwhich ELISA.The immune activity of the fusion protein was verified by Western blotting,and its molecular weight was about 51.76?10~(3),very close to the expected value.Conclusion The hB7-H1-Fc chimeric molecule was successfully constructed and the expression of its functional fusion protein in mammalian CHO cells lays a foundation for further research on the role of B7-H1 in immune tolerance,autoimmune diseases.

Objective To investigate the clinical value of modified BI?RADS classification by using the coronal plane of automatic breast volume scaner. Methods The total of 201 BI?RADS 3~5 classification of breast masses were retrospectively analyzed. All masses underwent conventional ultrasound and ABVS examination. Using BI? RADS classification standard terms to describe various information of breast masses, and record the coronal image of the masses on the complete interface echo, convergence sign, angle, burr, which classified BI?RADS ultimately. Results The coronal plane of convergence sign, complete interface echo, angulation and burr were significantly different between benign and malignant tumors (P<0.0001). The sensitivity of ABVS convergent sign in diagnosing breast malignant tumors was 68.2%, specificity was 93.4% and accuracy was 82%. The conventional ultrasound combined with the coronal feature of ABVS modified by BI?RADS classification showed that 3 kinds of malignant rate reduced from 8.5%to 3.2%. The rate of malignant 4a decreased from 25.2%to 12.1%and the rate of malignant 5 increased from 94.2% to 98%. Conclusion The convergence sign of ABVS can be used as a significant independent predictor of breast malignant tumors;ultrasound combined with ABVS is helpful to improve the accuracy of ultrasound BI?RADS classification.

BACKGROUND:Fentanyl transdermal system has analgesic effect similar to oral sustained-release morphine and has been widely used in advanced cancer pain management in several years. However, recent literatures about some serious adverse events associated with fentanyl transdermal system have been published, and the long-term safety of fentanyl transdermal system treatment is stil chalenged. OBJECTIVE:To observe the long-term clinical efficacy and safety of fentanyl transdermal system for pain management in patients with advanced cancer. METHODS:A total of 309 patients with advanced cancer pain were enroled, including 166 females and 143 males. The age ranged from 26-72 years old. Patients received oral sustained-release morphine for 2 weeks, and then were subject to fentanyl transdermal system for pain management at the 3rd week until the 12th week. A prospective study with self-contrast method was conducted to compare the analgesic effects of these two drugs, as wel as patient's acceptability, adverse events and toxicity in the administration course. RESULTS AND CONCLUSION: Stable pain relief was harvested throughout the oral administration of both sustained-release morphine and fentanyl transdermal system therapy. During the use of oral sustained-release morphine, adverse reactions appearing in sequence were constipation, nausea, fatigue and anorexia. After converting to fentanyl transdermal system, the symptoms of constipation (χ2=5.22,P=0.02) and nausea (χ2=4.38, P=0.04) significantly reduced, and vomiting was abated but showed no significant difference (χ2=2.7,P=0.10). 2.3% of patients had skin reactions to the patches, and regressed after replacing the patch area. Skin reactions were aleviated at 2-10 weeks after oral administration of fentanyl transdermal system. Some uncommon adverse events including headache, diarrhea, dyspnoea, excessive sweating or other symptoms often occurred at the time of the initial dosage increase. Preference or strong preference for fentanyl transdermal system in comparison to previous oral sustained-release morphine was reported by 91% of patients. These results demonstrate that fentanyl transdermal patches can provide stable pain relief for advanced cancer patients with good acceptability, and mitigate the incidence of adverse events due to oral drugs.

Objective To investigate the relationship between stent thrombosis and obstructive sleep apnea-hypopnea syndrome (OSAHS) in patients after coronary artery stent implantation (CASI). Methods One hundred and seventeen patients were rolled after CASI, of which 29 suffering from OSAHS were enrolled into the research group, 88 patients without OSAHS were enrolled into the control group. The incidence rate of stent thrombosis between the above two groups was analyzed. Results Three patients suffered stent thrombosis in the research group, with the incidence rate of 10.3%,which was significantly higher than that in the control group, with the incidence rate of 1.10%(P = 0.047). Conclusions After CASI, patients with OSAHS had higher risk of stent thrombosis compared with patients without OSAHS. OSHAS should be considered as a risk factor for stent thrombosis.

Objective To determine the baseline echocardiographic characteristics and the time course and degree of recovery of left ventricular (LV) systolic dysfunction in patients with tachycardia-induced cardiomyopathy ( TCM ) . Methods Seven hundred and fifteen patients received radiofrequency cather ablation ( RFCA ) for tachycardiarrhymias from July 2010 to July 2013 were screened in this study. Only 33 patients with reduced left ventricular ejection fraction (LVEF) (LVEF<50% and improved≥15%) were diagnosed with tachycardia-induced cardiomyopathy and were included in the study. Patients with early improvement (over 25%increase in LVEF at 1-week follow-up compared to the baseline ) were enrolled in the improved group , and the rest patients were enrolled in the improved group. All Patients received transthoracic echocardiography for LV size and function detection at 1 week and at 3,6,12 months follow-up. Results The average baseline of the LV end-diastolic diameter, and the LVEF were (55 ± 10.7)mm and (38 ± 4.6)%, repectively. Early improvement ( over 25%increase in LVEF at 1-week follow-up compared to the baseline ) in the improved group was observed in 16 patients. Patients with early improvement had higher LVEF at 12-month follow-up compared to the patients without early improvement [(69.2± 4.2)% vs (58.1 ± 6.9)%, P < 0.001]. Conclusions RFCA is proved to be a relatively safe and effecient treatment method. Atrial fibrillation related to TCM , rhythm control is superior to the rate control. The early improvement in LVEF may potentially predict the complete reversibility of LV systolic dysfunction.

Objective To investigate hemorrhagic transformation (HT) and outcomes in acute ischemic stroke.Methods The demographics,vascular risk factors,imaging and other clinical data in patients with acute ischemic acute were collected retrospectively and compared.Using the susceptibility weighted imaging (SWI) to diagnose HT,and the patients were divided into either a HT group or a non-HT group.The modified Rankin scale was used to evaluate the clinical outcomes.Multivariate logistic regression analysis was used to determine the independent risk factors for HT and poor outcome in HT patients.Results A total of 96 patients with acute ischemic stroke were enrolled and 34 of them had HT (35.4％).The age (66.21 ± 7.04 years vs.61.21 ±13.42 years; t =2.020,P=0.046) and infarct volume (3.88 ±2.20 cm3 vs.1.96 ± 1.37 cm3; t =5.67,P=0.001) in the HT group were significantly older or larger than those in the non-HT group.The proportions of hypertension (58.8％ vs.30.6％;x2 =7.228,P=0.007),diabetes (29.4％ vs.6.5％;x2 =9.293,P=0.002),atrial fibrillation (35.3％ vs.3.2％;x2=18.128,P=0.000),and cardiogenic cerebral embolism (35.3％ vs.3.2％ ; P =0.000) were significantly higher than those in the non-HT group,while the proportion of small arterial occlusive stroke was significantly lower than that in the non-HT group (38.2％ vs.62.9％ ;P =0.032).Multivariate logistic regression analysis showed that age (odds ratio [OR] 1.168,95％ confidence interval [CI] 1.059-3.412; P =0.021),infarct volume (OR 3.461,95 ％ CI 1.317-6.270; P =0.044) and atrial fibrillation (OR 1.284,95％ CI 1.117-2.903; P =0.015) were the independent risk factors for HT.In the HT patients,age (69.46 ±7.17 years vs.64.19 ±6.31 years; t =2.248,P =0.032) in the poor outcome group was significantly older than that in the good outcome group.The proportions of hypertension (84.6％ vs.42.9％ ;x2 =781,P =0.016),diabetes (50.0％ vs.14.3％ ;x2 =6.053,P =0.014),cardiogenic cerebral embolism (61.5％ vs.19.0％ ; P =0.025) and hematoma HT (76.9％ vs.19.0％ ;x2 =11.104,P =0.001) were significantly higher than those in the good outcome group.Multivariate logistic regression analysis showed that the diabetes (OR 2.151,95％ CI 1.179-3.218; P =0.023),atrial fibrillation (OR 4.136,95％ CI1.010 to 8.413; P =0.046) and hernatoma HT (OR 2.134,95％ CI 1.219-4.452; P =0.039) were the independent risk factors for the poor outcomes of HT patients at 3 months after symptom onset.Conelusions The incidence of HT in patients with acute ischemic stroke was 35.4％.Age,infarct volume and atrial fibrillation were the independent risk factors for HT,and diabetes,atrial fibrillation and hematoma HT were the independent risk factors for the poor outcomes in HT patients at 3 months after symptom onset.