1.Cloning and expression of MxA gene from chickens in E.coli and identification of biological activity of MxA
Lei CHEN ; Guotuo JIANG ; Weishan CHANG
Chinese Journal of Immunology 2000;0(11):-
Objective:To clone chickens MxA gene, construct its recombinant expression plasmid and induce the expression of fusion protein using a prokaryotic expression system.Methods:The MxA gene fragment was amplified by RT-PCR from CEF cells and subcloned into the pMD18-T vector, filtrated the positive clone and reclaimed the MxA.Subcloned the MxA into the prokaryotic expression plasmid pGEX-6p-1. After recombinant plasmid was induced by IPTG, the expressed proteins were analyzed by SDS-PAGE, the NDV intervence experiment and VSV-CEF restrain experiment.Results:The sequence of MxA gene amplified by RT-PCR was the same as the sequence in gene map of Genebank; SDS-PAGE, the NDV intervence experiment and VSV-CEF restrain experiment showed that a protein was expressed, the molecular weight of this protein was 45 000, which was the same as the fusion protein GST-MxA.Conclusion:The MxA is cloned and its recombinant expression plasmid is constructed successfully.The fusion protein GST-MxA is successfully expressed in the prokaryotic expression system E.coli DH5? induced by IPTG. This research lay a foundation for further studying on ints antiviral effects and exploring new way of antiviral medication.
2.The analysis of clinical features of 140 cases with primary hyperparathyroidism
Xiao'ai YAO ; Hong CHANG ; Tao JIANG ; Lei XIU ; Zhen WEN
Chinese Journal of Clinical Oncology 2016;43(23):1035-1039
Objective:To compare the clinical characteristics in primary hyper(-) parathyroid hormone (PHPT) of the different patholog-ic types. Methods:Clinical data of 140 patients with PHPT proved by operation and pathology during January 2010 to June 2016 were retrospectively analyzed. Results:A total of 140 PHPT patients, including 13 (9.29%) cases of parathyroid carcinoma (PC), 27 (19.29%) cases of parathyroid hyperplasia (PH), and 100 (71.43%) cases of parathyroid adenoma (PA). The duration of the PC group was longer than the PH group and the duration of the parathyroid adenoma (PH) group was longer than the PA group (P<0.05). The percentage of young patients with PC was higher than in the other two groups (P=0.003). The diameters of the PC group were larger than those of the other two groups, and those of the PA group were larger than those of the PH groups (P<0.05). Blood calcium, parathyroid hor-mone (PTH), AKP, fasting blood glucose (FBG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamete transpepti-dase (GGT), blood urine nitrogen (BUN), creatine (CRE), urinary calcium, and phosphorus of the PC group were higher than in the oth-er two groups (P<0.05). The blood calcium, PTH, alkaline phosphatase (AKP), urinary calcium of the PH group were lower than those in PA group (P<0.05). The proportion of ostalgia was 46.15%(6/13), 44.44%(12/27), and 49.00%(49/100). No statistical difference was observed (P>0.05). The postoperative calcium level of PC group was lowest (P<0.001), and the highest was of PTH (P<0.001). The pro-portions of clinical manifestation of the urinary system, digestive system, and nervous system in the PC group were 76.92%(10/13), 76.92%(10/13), and 15.38%(2/13), respectively, and these values were the highest in the three groups (P<0.05). The proportion of the clinical manifestation of the urinary system of the PH group was higher than that of the PA group. The fracture rate (30.77%, 4/13) and constipation rate (38.46%, 5/13) of the PC group were the highest among the three groups (P<0.05). Conclusion:The duration of patients with PC was the longest among the three groups. The percentage of young patients with PC was the highest. The abnormal parathyroid glands in the PC group were the heaviest. The PC group exhibited the lowest postoperative calcium level and the highest PTH level. The biochemistry and clinical manifestations of PC were obvious.
3.Research progress of integrin in tissue repair
Lei CUI ; Chang LIU ; Yanchao JI ; Jinsong LI ; Song XUE ; Long JIANG
International Journal of Surgery 2013;40(10):674-677
As a transmembrane glycoprotein receptor family,integrin family mainly mediats cell-matrix interactions,participates in the function,through the unique bi-directional signal transduction pathways,played an important role in tissue repair process.The purpose of this article is to summarize the role of integrin family in the repair of tissue damage and sum up the current research progress.
4.Hemocompatible coating of expanded polytetrafluoroethylene vascular graft based on cross-linked alginate/recombinant hirudin multilayer
Chang JIANG ; Xiuli YUE ; Fang MA ; Meng LIU ; Lei XING ; Nanqi REN
Chinese Journal of Tissue Engineering Research 2008;12(19):3765-3769
BACKGROUND: Thrombus formation and neointimal hyperplasia limit its use for revascularization of small-caliber vessels (<6mm diameter) in the coronary or peripheral circulation.OBJECTIVE: To improve hemocompatibility, the luminal surface of a small diameter expanded polytetrafluoroethylene (Eptfe) vascular prosthesis was modified with alginate and recombinant hirudin.DESIGN: Observational experiment.SETTING: This study was performed in Nanomedicine and Biosensor Laboratory, Bio-X Center, Harbin Institute of Technology from Marcy 2006 to June 2007.MATERIALS: The GORE-TEX Epife vascular grafts (W. L Gore & Associates, Inc., Flagstaff, AZ) used in this study were 4mm in internal diameter. rHir was obtained from Calbiochem, Germany. Sodium alginate (also called alginic acid sodium salt; medium viscosity) was purchased from Sigma.METHODS: A p-diazonium diphenyl amine polymer (PA) was used as an interlayer between alginate and recombinant hirudin (rHir). The diazonium moieties were capable of covalently coupling with electron-rich aromatic systems such as histidine and tyrosine residues of hirudin. No need for chemical pretreatment took all advantage by preserving the bulk properties with almost no effect on stability and elasticity of the Eptfe vascular graft. rHir amount on Epife surface Was determined by the Micro BCA (Bicinchoninic acid) Protein Assay kit.MAIN OUTCOME MEASURES: ① Determination rHir amount on Eptfe surface; ② Static water contact angles; ③ Attenuated total reflectance fourier transform infrared spectroscopy (ATR-FTIR) was employed to confirm the change taking place in the Eptfe graft surface modification process; ④ Characterization of the surface morphology and platelet adhesion by SEM; ⑤ APTT and PT; ⑥ Percent hemolysis.RESULTS: ① The amount of rHir adsorbed onto the Eptfe vascular was deduced to be 16.35 μg/cm2. ② Surface analysis ATR-FTTR revealed the presence of new functional groups on the modified graft surfaces. ③ The water contact angle of the modified graft surface decreased. ④ The longer APTT and PT value lower than 5% hemolysis level and dramatically decrease of platelet adhesion assay showed that rHir modified graft had great improved blood compatibility.CONCLUS10N: Cross-linked Alg/rHir onto Eptfe can improve luminal surface and hemocompatibility.
5.Colonization characteristics of endophytic bacteria NJ13 in Panax ginseng and its biocontrol efficiency against Alternaria leaf spot of ginseng.
Chang-Qing CHEN ; Tong LI ; Xin-Lian LI ; Yun JIANG ; Lei TIAN ; Peng XU
China Journal of Chinese Materia Medica 2014;39(10):1782-1787
To reveal the colonization characteristics in host of endophytic biocontrol bacteria NJ13 isolated from Panax ginseng, this study obtained the marked strain NJ13-R which was double antibiotic resistant to rifampicin and streptomycin through enhancing the method of inducing antibiotic. The colonization characteristics in ginseng and its biocontrol efficiency against Alternaria spot of ginseng in the field were studied. The results showed that the strain could colonize in root, stem and leaf of ginseng and the colonization amount was positive correlated with inoculation concentration. Meanwhile, the strain could infect and then transfer in different tissues of ginseng The colonization amount of strain in roots and leaves of ginseng increased first and then decreased. However, the tendency of colonization amount of strain in stems was ascend at first and then descend slowly, and was more than that in roots and leaves along with time, which had a preference to specific tissue of its host. In field experiment, the endophytic bacteria NJ13 was proved to be effective in controlling Alternaria leaf spot of ginseng. The biocontrol efficiency of fermentation broth at the concentration of 0.76 x 10(8) cfu x mL(-1) reached 75.62%, which was close to the controlling level (73.06%) of 0.67 mg x L(-1) 50% cyprodinil WG.
Alternaria
;
physiology
;
Antibiosis
;
Bacillus
;
growth & development
;
isolation & purification
;
physiology
;
Endophytes
;
growth & development
;
isolation & purification
;
physiology
;
Panax
;
growth & development
;
microbiology
;
Plant Diseases
;
microbiology
6.Utility of different body composition for the predicting myocardial ischemia in the type 2 diabetes mellitus (T2DM) patients
Lei JIANG ; Xianbo ZHANG ; Mingxiao SUN ; Dongni YU ; Lijuan WANG ; Shanshan REN ; Bo CHENG ; Jing WANG ; Xinmiao CHANG
Chinese Journal of Health Management 2016;10(5):361-365
Objective To explore the utility of different body composition for the prediction of myocardial ischemia in exercise in type 2 diabetes mellitus (T2DM) patients. Methods T2DM outpatients were selected from Jul. 2013 to Nov. 2014 (male 83, female 57) to measure body composition and clinical biochemical parameters, and divided them into two groups according to treadmill exercise testing results. Group A cases were positive (27 patients) and Group B cases were negative (113 patients). SPSS19.0 was used to make statistical analysis. Results (1) The age, gender, diabetes mellitus (DM) duration, medical history of hypertension and dyslipidemia, body mass index (BMI), waist circumference, the waist-hip fat ratio of the two groups had no significant difference (P>0.05). (2) The fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C) between the two groups had no significant difference. Compared with group B, the low density lipoprotein cholesterol (LDL-C) of group A [(2.48 ± 0.75) mmol/L vs. (2.81 ± 0.77) mmol/L, t=2.009, P<0.05] decreased significantly. (3) Overall body fat percentage, skeletal muscle weight of the two groups had no significant difference. Compared with group B, the waist-hip fat ratio (0.93±0.06 vs. 0.88±0.08, t=-2.790, P<0.01) andvisceral fat area [(99.47 ± 32.84) cm2 vs. (81.10 ± 25.47) cm2, t=-2.822, P<0.01] of group A increased significantly, while this difference mainly existed in male patients visceral fat area [(100.99±39.33) cm2 vs. (79.91 ± 27.09) cm2, t=-2.231, P<0.05], and waist-hip fat ratio was different in female patients of the two groups (0.91±0.06 vs. 0.85±0.09, t=-2.043, P<0.05) . Conclusion Abdominal obesity, especially visceral fat accumulation had close correlation with myocardial ischemia in exercise in T2DM patients. Waist-hip fat ratio and visceral fat area may be more effective indicators of myocardial ischemia in exercise in T 2DM patients than BMI.
7.Mahuang Zhisou Capsule in treatment of acute upper respiratory tract infection of external wind-cold syndrome: a multi-center, randomized controlled, and double-blind trial
Ying ZHANG ; Jing CHANG ; Ruiming ZHANG ; Liangli LIU ; Fengsen LI ; Xinyue JIANG ; Lei WANG ; Bin MAO ; Tingqian LI
Journal of Integrative Medicine 2008;6(6):581-5
OBJECTIVE: To evaluate the efficacy and safety of Mahuang Zhisou Capsule (MHZSC), a compound traditional Chinese herbal medicine, in treatment of acute upper respiratory tract infection of wind-cold syndrome. METHODS: A multi-center, double-blind, double dummy, randomized controlled trial was conducted. A total of 240 patients were randomized into treatment group (120 cases, treated with MHZSC) and control group (120 cases, treated with Mahuang Zhisou Pill). The therapeutic course was 3 days. RESULTS: Seven cases were excluded and three cases lost to follow-up. Two hundred and thirty-three cases entered intention-to-treat (ITT) analysis and 230 cases fitted in per-protocol population set (PPS) analysis. After treatment, the therapeutic effect of upper respiratory tract infection was calculated by ITT and PPS analysis respectively. The significant response rate and total response rate in the treatment group were 78.07% and 98.25% (ITT and PPS), while the significant response rates in the control group were 70.59% (ITT) and 70.69%(PPS), and the total response rates were 95.80% (ITT) and 95.69% (PPS) respectively. There were no significant differences in clinical efficacy between the two groups (P>0.05). CONCLUSION: MHZSC is effective and safe in treatment of acute upper respiratory tract infection of external wind-cold syndrome.
8.Effect of collagens and growth factors (TGFß1, PDGF-BB) on cytoskeletal reorganization and cell migration in hepatic stellate cells.
Lei LI ; Wei JIANG ; Ji-yao WANG ; Chang-qing YANG
Chinese Journal of Hepatology 2010;18(8):581-584
OBJECTIVETo evaluate the effects of collagens and growth factors TGFß1 and PDGF-BB on the cytoskeletal components and migration in cultured rat hepatic stellate cells (HSCs).
METHODSPrimary rat hepatic stellate cells were isolated and cultured. A Transwell Chamber system was used to observe the changes of serum starved HSCs haptotactic migration (direct stimulation) and chemotactic migration (indirect stimulation) after collagens and growth factors treatment. Changes in actin cytoskeletal organization were visualized by fluorescence staining using FITC-labeled phalloidin and the fluorescence images were recorded using confocal microscopy.
RESULTSTGFß1 enhanced significantly the motility of primary HSCs at 5 ng/ml: for haptotactic migration cells: 131.37+/-3.15 vs 102.93+/-1.01, F=40.84, P<0.05; for chemotactic migration cells: 210.17+/-1.78 vs 102.93+/-1.01, F=64.53, P<0.05. PDGF-BB enhanced significantly the motility of primary HSCs at 10 ng/ml (haptotactic migration cells: 203.67+/-7.54 vs 102.93+/-1.01, F=40.90, P<0.05; chemotactic migration cells: 319.56+/-11.71 vs 102.93+/-1.01, F=54.57, P<0.05); Both chemotactic and haptotactic stimuli with 100 microg/ml collagen type I significantly increased HSCs migration: for haptotactic migration cells: 127.20+/-6.47 VS 102.93+/-1.01, F=41.01, P is less than 0.05; for chemotactic migration cells: 201.52+/-11.28 vs 102.93+/-1.01, F=36.49, P<0.05; however, collagen type IV had no effect on HSCs migration. Serum-starved, untreated cells had a rounded-up morphology. PDGF-BB and TGFß1 induced a rapid morphological change concomitant with a robust reorganization of actin cytoskeleton in HSCs.
CONCLUSIONSCollagen type I, PDGF-BB and TGFß1 could induce cell migration in HSCs, but collagen type IV has no such effect. PDGF-BB and TGFß1 could induce the actin cytoskeleton reorganization in HSCs.
Actins ; drug effects ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Collagen Type I ; pharmacology ; Collagen Type IV ; pharmacology ; Cytoskeleton ; drug effects ; Hepatic Stellate Cells ; cytology ; drug effects ; Male ; Platelet-Derived Growth Factor ; pharmacology ; Proto-Oncogene Proteins c-sis ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; pharmacology
9.Screening and identification of indoleacetic acid producing endophytic bacterium in Panax ginseng.
Yun JIANG ; Lei TIAN ; Chang-qing CHEN ; Guan-jun ZHANG ; Tong LI ; Jing-xiu CHEN ; Xue WANG
China Journal of Chinese Materia Medica 2015;40(2):213-217
Endophytic bacteria which was producing indoleacetic acid was screened from Panax ginseng by using the Salkowski method. The active strain was also tested for its ability of nitrogen fixation by using the Ashby agar plates, the PKV plates and quantitative analysis of Mo-Sb-Ascrobiology acid colorimetry was used to measure its ability of phosphate solubilization, for its ability of potassium solubilization the silicate medium and flame spectrophotometry was used, for its ability of producing siderophores the method detecting CAS was used, for its ability of producing ACC deaminase the Alpha ketone butyric acid method was applied. And the effect on promoting growth of seed by active strain was tested. The results showed that the indoleacetic acid producing strain of JJ5-2 was obtained from 118 endophytes, which the content of indoleacetic acid was 10.2 mg x L(-1). The JJ5-2 strain also had characteristics of phosphate and potassium solubilization, nitrogen fixation, producing siderophores traits, and the promoting germination of ginseng seeds. The JJ5-2 strain was identified as Bacillus thuringiensis by analyzing morphology, physiological and biochemical properties and 16S rRNA gene sequences.
Bacteria
;
isolation & purification
;
metabolism
;
Endophytes
;
isolation & purification
;
metabolism
;
Indoleacetic Acids
;
metabolism
;
Panax
;
microbiology
10.Expression of recombinant human lysozyme-tachyplesin I (hLYZ-TP I) in Pichia pastoris and analysis of antibacterial activity.
Yu GAO ; Hong Lei ZHAO ; Xin FENG ; Rui Dong ZHAI ; Seng ZHU ; Chong Tao DU ; Chang Jiang SUN ; Lian Cheng LEI
Biomedical and Environmental Sciences 2013;26(4):319-322
Antimicrobial Cationic Peptides
;
biosynthesis
;
genetics
;
DNA-Binding Proteins
;
biosynthesis
;
genetics
;
Humans
;
Microbial Sensitivity Tests
;
Muramidase
;
biosynthesis
;
genetics
;
Peptides, Cyclic
;
biosynthesis
;
genetics
;
Pichia
;
metabolism
;
Recombinant Proteins
;
biosynthesis
;
genetics
;
isolation & purification