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.

Objective To investigate the compartmentalization of human immunodeficiency virus (HIV)infection and CD4~+ T lymphocytes between gastric mueosa and peripheral blood from patients with acquired immune deficiency syndrome(AIDS).Methods Thirty-five AIDS patients and 10 HIV seronegative subjects were enrolled in this study.The gastric mucosal biopsies were done by gastroscope and peripheral blood samples were collected.The digoxin labeled HIV-1 double-stranded cDNA probes for the long terminal repeat(LTR)and gag gene of HIV-1 genome were prepared by polymerase chain reaction(PCR).In situ hybridization was used to detect HIV infection in gastric mucosal tissues and peripheral blood mononuclear cells(PBMC)of AIDS patients.CD4~+ T lymphocytes were determined by immunohistochemistry method.The data were analyzed using t test.Results HIV positive rates were(1.67±1.48)% in gastric mucosa and(19.37±9.23)% in PBMC of AIDS patients who had not received highly active antiretroviral therapy(HAART).The HIV positive rates in gastric mucosa were not significantly different between AIDS patients without treatment and those treated with HAART(t=-0.996,t=-0.794,t=-0.461;P＞0.05).HIV positive rate on PBMC film preparation in patients who had received HAART for 1-4 years was (4.25±3.47)%,which was significantly lower than untreated group[(19.37±9.23)%](t=3.000,P＜0.05).The positive rate of CD4~+ T lymphocytes was(12.53±8.14)% in gastric mucosal mononuclear cells(MMC)and(19.00±9.55)% in PBMC of AIDS patient who had not received HAART.Even after received HAART for 1-4 years,the positive rate of CD4 ~+ T lymphocytes in MMC were(37.44±18.00)%,which was still lower than control group[(50.35±3.41)%](t=-4.620,P＜0.01);while the positive rate of CD4~+ T lymphocyte in PBMC was similar with that of control group(t=-2.094,P＞0.05).Conclusion The compartmentalization of HIV infection and CD4~+ T lymphocyte counts exists between gastric mucosa and peripheral blood from AIDS patients.

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.

OBJECTIVETo investigate human cytomegalovirus (HCMV) and Epstein-Barr virus-type 1 (EBV-1) in GCF and saliva during experimental gingivitis in Chinese young subjects and to evaluate the effect of the virus in the initial stage of gingival inflammation.

METHODSGCF of 14 and 45 and saliva without stimulating in 11 Chinese young males with healthy gingiva were collected at baseline (day 0), day 7, 14 and 21 after stopping oral hygiene and day7 after reestablishing oral hygiene (day 28). DNA of HCMV and EBV-1 were detected by nested-polymerase chain reaction (n-PCR) at the times mentioned above.

RESULTSHCMV was detected in GCF of 4 subjects at baseline, 4 subjects at day 7, 3 subjects at day 14 and 2 subjects at day 21 while the subjects were different. At day 28 HCMV could not be detected. EBV-1 was not detectable in GCF during the experimental gingivitis. HCMV was detected in saliva in 4 subjects and EBV-1 was in 3 subjects. And there is no relationship between the detection of the herpesviruses and the clinical parameters as well.

CONCLUSIONWe suggest that HCMV and EBV-1 are not the important factors during the initial stage of gingival inflammation.

Adult ; Cytomegalovirus ; isolation & purification ; Gingival Crevicular Fluid ; virology ; Gingivitis ; virology ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Young Adult

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.