Formate dehydrogenase(FDH)coding gene was amplified from genomic DNA of Pichia pastoris by polymerase chain reaction, and the codon TAG(bases 649-651)was mutated to GAG using site-directed mutagenesis.The recombinant plasmid pET-FDH was con- structed by inserting the mutated DNA fragment into expression vector pET-22b(+),and transformed into E.coli BL21(DE3).FDH was expressed as a form of soluble prutein fused with 6?His tag at high level through IPTG induction.The amount of FDH was up to about 30% of the total cell protein.The cells-free crude extract was purified by one affinity chromatographic step,and resulting enzyme preparation revealed a specific activity of 6.45 U/mg.

Adenoviridae ; genetics ; Cell Line ; Genetic Vectors ; Humans ; Nitric Oxide Synthase Type III ; genetics ; Virus Replication

Objective · To compare the short and medium-term efficiency and safety of 1470 nm laser with foam sclerotherapy with ligation of great saphenous vein and 808 nm laser with foam sclerotherapy to treat varicose veins of lower extremity. Methods · Seventy-seven hospitalized patients patients and twenty-one day-surgery patients were retrospectively analyzed. Results · There was no obvious difference in the closure of great saphenous vein for short to medium-term efficiency between the day-surgery group and hospitalized group. The postoperative complications and recurrence rate of day-surgery group was not higher than that of the hospitalized group, but the degree of perioperative comfort, economic cost and the time returning to normal life of day-surgery group were superior to that of the hospitalized group. Conclusion · 1470 nm laser with foam sclerotherapy with ligation of great saphenous vein and 808 nm laser with foam sclerotherapy to treat varicose veins of lower extremity had similar clinical effect, but the degree of safety and comfort was higher and trauma was less. The day surgery with 1470 nm laser to treat varicose vein of lower extremity could also simplify the procedure of hospitalization and reduce the cost of health care, which could be a new option for patients.

Objective To investigate the molecular mechanism of the effect of CYP2C19 polymorphism on the efficacy of clopidogrel in patients with stable coronary artery disease (SCAD).Methods A total of 208 patients with coronary artery disease undergoing percutaneous coronary intervention (PCI) were included.The CYP2C19 variant alleles were detected by gene sequencing.Platelet reactivity was assessed by thrombelastograph at 24 h after 300 mg clopidogrel loading.P2Y12-Gi signaling was measured by flow cytometric analysis of vasodilator-stimulated phosphorylation-platelet reactivity index (VASP-PRI) and Akt phosphorylation (P-Akt) index.Results Among 208 patients,40.9％ were classified as CYP2C19 * 1/* 1 (non-carriers) with no loss-of-function (LOF),44.7％ as CYP2C19 * 1/* 2 or CYP2C19 * 1/* 3 (one LOF carriers) and 14.4％ as CYP2C19 * 2/*2 or CYP2C19 * 2/* 3 (two LOF carriers).Both postclopidogrel platelet aggregation and VASP-PRI increased by the presence of one LOF allele,and were even more pronounced with the presence of two LOF alleles.In contrast,P-Akt index only increased in two LOF carriers.VASP-PRI was positively associated with postclopidogrel platelet aggregation (r =0.661,P＜0.001),but not with P-Akt index.Conclusions The two CYP2C19 LOF carriage was associated with more active state of P2Y12-Gi signaling in postclopidogrel platelet in Chinese patients with SCAD.

Objective Meta-analysis the efficacy and safety of Exenatide and insulin therapy oral hypoglycemic drugs effect of obesity with type 2 diabetes .Methods According to the research purpose to set up the screening of related literature and exclusion criteria; formulatethe searching strategy, through PubMed、the Chinese Biological Medicine Datebase(CBM)、 CNKI、Wanfang Data Knowledge Service Platform, VIP to retrieve all theliterature selection of efficacy and safety by Exenatide oral hypoglycemic drugs and insulin therapy of obese type 2 diabetes mellitus.Choose met inclusion exclusion criteria, the complete data information randomized controlled trial (RCT) as the research object; Apply to the international commonly used Jadad score method to evaluate quality included in the test; To process the relevant data in the test ; Apply the ReviewManager 5.1 software to analysis the extracted research data.Analysis the results and put forward conclusions.Results Participants included 11 RCT , meta analysis results showed that compared with the Exenatide, in terms of reducing fasting glucose ,insulin effect more apparent [MD = 0.35, 95％CI: (0.11, 0.59), P = 0.004)]. In control effect of glycosylated hemoglobin, there was no statistically significant difference[MD=-0.04 ,95％CI:(-0.20,0.11), P=0.58],between Exenatide and insulin. Compared with the insulin, Exenatide reduce BMI more apparent[MD=-2.77,(95％CI: -3.34,-2.20),P<0.00001]; Compared with the insulin, Exenatide reduce insulin resistance index, the effect is more obvious[MD=-1.67,95％CI:(-1.93,-1.41), P<0.00001]; Adverse reaction in the process of treatment, the insulin is more likely to lead to hypoglycemia, [OR = 0.32, 95％ CI: 0.19, 0.54), P<0.0001]; While Exenatide are more likely to lead to gastrointestinal adverse reaction [OR = 4.04, 95％ CI: 2.35, 6.93), P<0.00001).Conclusion According to the Meta-analysis: Exenatide can be used in the treatment of oral hypoglycemic drugs of adult obesity with type 2 diabetes, and obvious effects of treatment of insulin resistance, long-term results still needs a large number of samples of high quality RCT to verify.

The theory of four properties (Qi) and five tastes (Wei) is the core of the property theory of Chinese materia medica. It is known that Qi and Wei are associated with the pharmacological effects (Xiao) of herbs. This study took records of all 365 Chinese herbs in Shennong's Classic of Materia Medica (Shennong Ben Cao Jing) as the data resource and established a three-dimensional data cube, in the purpose of finding out and analyzing the frequent patterns and valued association rules of Qi, Wei and Xiao based on Apriori algorithm. The results of this study may give rise to innovative ideas and methods in research of traditional Chinese materia medica.

Hydrocortisone-induced yang-deficiency animal model has now become the generally accepted model of yang deficiency. However, assessing the most appropriate dose of hydrocortisone is a long-term challenge. For analyzing the modeling dose, the authors have built several kinds of yang-deficiency models induced by hydrocortisone at different doses, and analyzed the experimental data with various mathematical statistical methods. In order to discuss the effects of the modeling dose on the basis of previous research, the authors introduced Ridit analysis.

Objective To investigate histopathological changes and expression of integrin ?1 of sternomastoid muscle,and probe the mechanism and significance during disease process in congenital muscular torticollis(CMT).Methods Histopathological changes of sternomastoid muscle section stained with HE and Gomori silver staining were observed and the expression of integrein ?1 with immunohistochemistry was detected,and the expressive quantity and distribution with image analysis system was quantitive analyzed.Results 1.With light microscopy observation,the results showed that the fibrous degeneration of sternomastoid muscle could be summed up 2 kinds: A category displayed the myocytes atrophyed,and there were lots of connective tissue hyperplasy around myocytes,and the direction of fibrous arrangement was disordered,meanwhile there were lots of vessels and nervers hyperplasy,and eventually the myocytes shrank back and disappeared.B category displayed that the structure of cross striation or sarcomere disappeared or changed,and myocytes could maintained the outline and the sarcolemma were integrated,and then fibrous pathological changes of myocyte took place,and there were lots of fibroblast-like that had much more enations between fiber bundles.With Gomori silver staining,the major changes of fibrotic sternomastoid muscle showed that there were lots of collagenous fibers hyperplasy.The arrangements of collagenous fibers were disordered in A category and were well-arranged in B ca-tegory.2.With immunohistochemistry,the results showed the expression of integrin ?1 was weak positive in normal control group(125.7?5.167).In diseased groups,the results showed 3 different extents:the expression of integrin ?1 displayed stronger positive in A category myocytes(30.15?6.543),and the level of expression was significantly different from normal controls(P0.05).Conclusions The fibrous pathological changes of sternomastoid muscle are a complicated and gradually process,which may has different mode,and ingetrin ?1 may participated the process of pathological changes.

Objective:To compare the efficacy of bilateral and unilateral metallic stenting in the treatment of unresectable malignant hilar biliary obstruction.Methods:From January 2012 to December 2018, a total of 300 patients with malignant hilar biliary obstruction (Bismuth type Ⅱ-Ⅳ) were treated with endoscopic metallic stenting in Oriental Hepatobiliary Surgery Hospital. Ninety-four patients with bilateral metallic stenting (bilateral metallic stent group) and 94 patients with unilateral metallic stenting (unilateral metallic stent group) were matched and analyzed by propensity score matching. The clinical success rate, the total number of interventions, the stent patency and overall survival time were compared between the two groups.Results:The clinical success rate in the bilateral metallic stent group was significantly higher than that of the unilateral metallic stent group [98.9% (93/94) VS 78.7% (74/94), χ2=19.352, P<0.001], and the mean number of intervention in the bilateral metallic stent group was significantly less than that of the unilateral metallic stent group (1.2±0.5 times VS 1.7±0.9 times, t=-4.345, P<0.001). The stent patency time in the bilateral metallic stent group was significantly longer than that of the other group [10.0 (8.0, 12.1) months VS 5.7 (5.2, 6.3) months, χ2=19.789, P<0.001]. The median survival time of patients in the bilateral group was numerically longer than that in the unilateral group, but did not reach statistical significance [7.6 (6.3, 8.9) months VS 4.6 (3.7, 5.7) months, χ2=3.628, P=0.057]. Conclusion:Endoscopic bilateral metallic stenting is superior to unilateral metallic stenting in the clinical management of malignant hilar biliary obstruction unsuitable for surgery.

Objective:To investigate the feasibility and efficacy of endoscopic placement of transpapillary parallel-style bilateral metal stents for malignant hilar biliary stricture.Methods:A total of 55 patients with malignant hilar biliary stricture (Bismuth Ⅱ-Ⅳ) were treated with an improved endoscopic drainage technique (endoscopic transpapillary parallel-style bilateral metal stenting) from January 2012 to December 2018 in Shanghai Oriental Hepatobiliary Surgery Hospital. The technical success rate, clinical success rate, incidence of complications, success rate of endoscopic re-intervention, stent patency and overall survival were analyzed.Results:The technical success rate of endoscopic placement of bilateral metal stents was 96.4% (53/55), the clinical success rate was 96.2% (51/53), and the early complication incidence was 13.2% (7/53). The median stent patency was 9.2 months (95% CI: 8.0-10.3 months), the success rate of endoscopic re-intervention of bilateral biliary systems was 92.3% (12/13), and the median overall survival was 6.7 months (95% CI: 4.7-8.8 months). Conclusion:This modified technique of bilateral metal stent placement is safe and effective for malignant hilar biliary stricture that is unsuitable for surgery.