Objective:To investigate the clinical efficacy and safety of antiplatelet combined with anticoagulation therapy for restenosis prophylaxis after stent placement in diabetic patients with leg atherosclerosis obliterans(LASO). Methods:83 diabetic LASO patients with stent placement were collected and randomly divided into the treatment group(42 cases)and the control group (41 cases)with 49 sick limbs in each. The control group was treated with clopidogrel(75mg,for 1 year)plus bayaspirin(100 mg),while the treatment group was treated with low molecular heparin(withdrawal when INR = 2. 0- 3. 0)plus warfarin(for 6 months)additionally. All cases were followed up for 12 months and the efficacy and safety were compared and evaluated. Results:After therapy,there were no differences in FPG,GHbA1C,TC and LDL-C at each time point between the two groups(P >0. 05). Compared with that in the control group,MLD in the treatment group was improved obviously(P < 0. 05)in the 12th month after the surgery,while restenosis rate and level of D-D in plasma were declined significantly(P < 0. 05). And there was no difference in bleeding rate between the two groups(P > 0. 05). Conclusion:Compared with antiplatelet therapy,antiplatelet combined with anticoagulation therapy for restenosis prophylaxis after stent placement in diabetic LASO patients is more effective and safer,which is worthy of clinical promotion.

Objective To explore the method and efficiency of percutaneous transluminal septal tunnet myocardial ablation(PTSTMA) in treatment of 26 cases hypertrophic obstructive cardiomyopathy (HOCM) who were not suitable for conventional technology.Methods Firstly,we used a monorail Balloon which was slightly bigger than the interventricular septal branch of coronary artery and dilated it until posterior septal.After that,an OTW Balloon with larger size than the monorail was used to dilate again until made aventricular septum tunnel.Then,some alcohol was injected and PTSTMA was performed.Finally,we did the other and/or another interventricular septal branch by above method until the left ventricular outflow tract pressure gradient (LVOTPG) reduced ≥50％.The clinical indexes of the 26 cases HOCM immediately pest-operation of PTSTMA were observed and the follow up data during short term and metaphase were analyzed.Results The LVOTPG reduced ≥50％ in the26 cases HOCM,immediately after PTSTMA,the LVOTPG reduced from (75.6 ±22.4)mm Hg to (21.4 ± 5.8) mm Hg (t =11.94,P ＜ 0.01).At three months after ablation,the thickness of septal myocardium reduced from (22.8 ± 5.8) ram before ablation to (16.8 ± 4.2) mm(t =4.27,P ＜ 0.01),left atrium dimension reduced from(48.0 ±7.0) mm to (42.0 ±8.6) mm (t =2.76,P ＜0.01).Followed up 6.0to 60.0 months,the patients suffering from chest pain reduced from 14 cases before to 4 cases after the procedure(53.8％ (14/26) vs 15.4％ (4/26),x2 =8.49,P ＜ 0.01),the patients with expiratory dyspnea reduced from 26 cases to 5 cases(100％ (26/26) vs 19.2％ (5/26),x2 =35.22,P ＜ 0.01),NYHA functional class improved from (2.4 ± 0.6) to (1.4 ± 0.7) (t =5.53,P ＜ 0.01).Conclusion The PTSTMA was a supplemental method of PTSMA on treating HOCM,which was safe and useful during the short term and metaphase.

Objective To construct a fusion protein that used for treatment of resistance and palindromia in leukemia and studied its biological activity. Methods IL-3 and LP gene fragments were amplified by PCR. After enzymatic digestion and T4 ligation, the fusion gene was cloned into expression vector pAYZ. The product was purified by exchange chromatography and anti-Etag affinity chromatography. IL3-G4SLP fusion protein was analyzed by SDS-PAGE and Western blot. Protein biological activity was detected by FACS. Results The fusion protein was expressed as soluble protein by E.Coli 16C9. The protein expression level was about 1 mg/L, its purity was over 95 %, and the expression level was about 1 mg/L. The fusion protein can combined specificely with CD123 on leukemia stem cells. Conclusion Fusion protein IL-3-G4S-LP can target on leukemia stem cells and maybe as a potential drug used for treatment of resistance and palindromia in leukemia.

OBJECTIVE To investigate the inhibitory effect of eight lignan compounds of Fructus Schisandrae chinensis in vitro on carboxylesterase 2 (CES2) and to estimate the herb-drug interaction (HDI) risks of strong CES2 inhibitors selected from the above compounds. METHODS Fluorescein diacetate (FD) was employed as a specific fluorescent probe of CES2. The residual activity of CES2 was detected in human liver microsomes after the intervention with deoxyschizandrin, schisanhenol, schisantherin E, schisandrol A, schisandrol B, gomisin J, gomisin G, and gomisin O at 37℃ for 10 min, respectively. 1% DMSO served as control. Residual activity of CES2 was assessed with metabolite production of FD detected by fluorescent intensity, combined with IC50 values of the above compounds to predict HDI risks between lignans and CES2-metabolizing drugs. RESULTS Compared with control group, the activity of CES2 was significantly inhibited by deoxyschizandrin and schisanhenol (P<0.01), with IC50 values of 8.06 μmol · L- 1 and 8.91 μmol · L- 1, respectively. The other six lignans compounds exhibited mild inhibitory effect on CES2. HDI risk prediction of deoxyschizandrin or schisanhenol indicated that exposure of CES2-metabolizing drugs might increase 11.24 and 0.40 times, respectively. CONCLUSION Deoxyschizandrin and schisanhenol exhibit strong inhibitory effects against CES2 in vitro so that potential HDI risks should be taken into account during administration of drugs containing Fructus Schisandrae chinensis.

ObjectiveTo investigate the effect of α-Zearalanol(α-ZAL) on lipometabolism and hemorheology in ovariectomized(OVX) hyperlipidemia rabbits.Methods44 adult virgin female rabbits were divided into 5 groups,group A: normal control;group B: sham+CHO;group C: OVX+CHO;group D: OVX+CHO+17βE_2;group E: OVX+CHO+α-ZAL.Cholesterol(CHO) was fed to rabbits for 12 weeks.Before and after feeding CHO,the serum lipid(TC,TG,LDL-C,HDL-C) were measured;Blood viscosity,plasma viscosity,aggregation index of RBC(AIRC) and fibrinogen were also assayed respectively.ResultsThe serum levels of TC,TG and LDL-C in group B and E were significantly decreased compared with those in group C(P<0.05);the level of blood viscosity,plasma viscosity and AIRC platelet aggregation rate in group D and E were also significantly decreased compared with those in group C(P<0.05).Conclusionα-ZAL can improve vascular function through the adjustment of lipometabolism and hemorheology.

Objective To investigate the effectiveness and safety of preoperative intravenous administration of tranex-amic acid in the hemiarthroplasty treatment of elderly hip fracture. Methods Seventy patients who received hemiarthro-plasty due to femoral neck fracture (Garden Ⅲ/Ⅳ) in our hospital from January 2008 to December 2012 were analyzed retrospectively, of which 35 patients were given preoperative intravenous administration of tranexamic acid and 35 pa-tients were not. The patients'preoperative and postoperative hemoglobin and hematocrit values were collected. The to-tal blood loss amount was calculated through Gross equation and the postoperative drainage amount, number of people receiving blood transfusion, blood transfusion amount and occurrence of thrombus events were recorded. Results The total blood loss amount of the tranexamic acid group was (867.79±76.93) mL, which was significantly lower than (1207.07±403.83) mL of the control group, with statistically significant difference(P=0.036). The postoperative drainage amount was (305.67±103.68) mL, which was lower than the (393.00±66.29) mL of the control group, with statistically significant difference (P<0.01). The application of tranexamic acid decreased the blood transfusion rate from 42.86% to 20.00%(P=0.039). The complications of postoperative thrombus events did not increase (P=0.643). Conclusion In the treatment of elderly femoral neck fracture, the preoperative intravenous administration of tranexamic acid can effec-tively and safely reduce the blood loss amount and blood transfusion rate of hemiarthroplasty.

Objective To investigate the effectiveness of the affinity adsorption material developed by our team for the specific removal of exogenous endotoxin in the blood circulation. Methods Fifteen beagle dogs were intravenously injected with endotoxin to establish a dog model of endotoxemia, and then they were randomly divided into the treatment group(n=10)and the control group(n=5). The treatment group received an extracorporeal perfusion to remove the endotoxin using the self-made disposable hemoperfusion device,while the control group using routine perfusion device. The levels of endotoxin, tumor necrosis factor α(TNF-α), interleukin 1β(IL-1β), interleukin 6(IL-6)and interleukin 8 (IL-8)in the blood of the dogs were measured at the beginning and 120 min after hemoperfusion for 120 minutes. The vital signs of the dogs were monitored during the hemoperfusion. Results After successful establishment of the endotoxemia model,the level of endotoxin at the beginning of hemoperfusion in the treatment group and control group was 118.63 ± 27.98 EU/mL and 117.16 ± 22.95 EU/mL,respectively. After hemoperfusion for 120 min,it was 0.039 ± 0.01 EU/mL and 131.98 ± 7.01 EU/mL, showing a significant difference(P﹤0.05). The clearance rate of hemoperfusion in the treatment group was 94.07%. At the beginning of hemoperfusion, the levels of TNF-α, IL-1β, IL-6 and IL-8 in the treatment group were 1.53 ± 0.27 ng/mL,12.82 ± 1.66 ng/mL,54.77 ± 3.98 ng/mL and 0.25 ± 0.32 ng/mL, and the levels in the control group were 1.53 ± 0.06 ng/mL,13.05 ± 0.18 ng/mL,54.58 ± 0.19 ng/mL and 0.28 ± 0.06 ng/mL, respectively. After hemoperfusion for 120 min, the levels of TNF-α, IL-1β, IL-6 and IL-8 in the treatment group were 0.13 ± 0.06 ng/mL, 0.70 ± 0.36 ng/mL, 1.62 ± 0.80 ng/mL and 0.01 ± 0.00 ng/mL, respectively, and as for the control group,the levels were 2.26 ± 0.15 ng/mL,15.12 ± 0.18 ng/mL,62.54 ± 0.93 ng/mL and 0.73 ± 0.93 ng/mL. There were significant differences between the beginning and after perfusion for 120 min in those two groups(P< 0.05). Conclusions This affinity adsorption material can effectively remove endotoxin and the inflammatory mediators in the blood of experimental dogs,with a clearance rate of 94.07%.

Protein A and protein G are two well-defined immunoglobulin (Ig)-binding proteins (IBPs), which show affinity for specific sites on Ig of mammalian hosts. Protein A and protein G contained several highly homologous IgG-binding domains which had been demonstrated to have function to bind to IgG. Whether combinations of Ig-binding domains of various IBPs could produce useful novel binding properties remains interesting. We constructed a combinatorial phage library which displayed randomly-rearranged A, B, C, D and E domains of protein A, B2 and B3 domains of protein G. Four rounds molecular evolution of this library directed by all four human IgG subclasses respectively generated a common arrangement of D-C respectively which didn't exist in SpA. The dynamic loss of control phages and increase of the phages displaying two or more binding domains, especially the selective enrichment of D-C and strict selection of its linking peptides demonstrated the efficient molecular evolutions and the significance of the selected D-C arrangement. The phage binding assays confirmed that D-C possessed a binding advantage with four human IgG subclasses compared to SpA. In this work, a novel combination of Ig-binding domains, D-C, was obtained and presented the novel Ig binding properties which provided a novel candidate molecule for the purification, production and detection of IgG antibodies and a new approach for the further study of structures and functions of IBPs.
Amino Acid Sequence ; Antibody Specificity ; Bacterial Proteins ; immunology ; metabolism ; Binding Sites ; Binding, Competitive ; Evolution, Molecular ; Immunoglobulin G ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Sequence Alignment ; Staphylococcal Protein A ; immunology ; metabolism

We constructed a phage-displayed random mutation library of Tat38-61(51N/55N), for studying the molecular evolution screening of HIV-1 Tat38-61 epitope. We used primers containing the random nucleotide sequences, and introduced the random mutations at the sites of 51 and 55 amino acids coding sequences into full-length Tat sequences by overlapping PCR. With the randomly mutated full-length Tat as template, the Tat38-61(51N/55N) mutants which contained recognition sequences for the Xba I in both ends were amplified by PCR using the designed primers. The mutants were cloned into Xba I site in the phagemid vector pCANTAB5S, then the recombinants were transformed into E. coli TG1, a phage-displayed the random mutation library of Tat38-61(51N/55N) was constructed by the rescue of help virus M13KO7. The results showed that the library consisted of about 5.0 x 10(6) colonies and the phage library titer was 2.65 x 10(12) TU/mL. More than 56.50% colonies in the library were positive for insertion. Sequence analysis showed that the nucleotides encoding amino acids at the sites of 51 and 55 distributed randomly. The constructed mutation library could meet the requirements for the following molecular evolution screening, and might prepare the Tat mutants for the further study of new Tat vaccine candidates.
AIDS Vaccines ; immunology ; Escherichia coli ; genetics ; metabolism ; HIV-1 ; genetics ; Humans ; Mutation ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; tat Gene Products, Human Immunodeficiency Virus ; biosynthesis ; genetics ; immunology

Through bioinformatics predictions, we identified that GTF2I and FAT1 were downregulated in thyroid carcinoma (TC). Further, Pearson's correlation coefficient revealed a positive correlation between GTF2I expression and FAT1 expression. Therefore, we selected them for this present study, where the effects of bone marrow mesenchymal stem cell-derived EVs (BMSDs-EVs) enriched with GTF2I were evaluated on the epithelial-to-mesenchymal transition (EMT) and stemness maintenance in TC. The under-expression of GTF2I and FAT1 was validated in TC cell lines. Ectopically expressed GTF2I and FAT1 were found to augment malignant phenotypes of TC cells, EMT, and stemness maintenance. Mechanistic studies revealed that GTF2I bound to the promoter region of FAT1 and consequently upregulated its expression. MSC-EVs could shuttle GTF2I into TPC-1 cells, where GTF2I inhibited TC malignant phenotypes, EMT, and stemness maintenance by increasing the expression of FAT1 and facilitating the FAT1-mediated CDK4/FOXM1 downregulation. In vivo experiments confirmed that silencing of GTF2I accelerated tumor growth in nude mice. Taken together, our work suggests that GTF2I transferred by MSC-EVs confer antioncogenic effects through the FAT1/CDK4/FOXM1 axis and may be used as a promising biomarker for TC treatment.
Mice ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Mice, Nude ; Epithelial-Mesenchymal Transition ; Thyroid Neoplasms/pathology* ; Extracellular Vesicles/pathology* ; Mesenchymal Stem Cells ; Transcription Factors, TFIII/metabolism* ; Neoplastic Stem Cells/pathology*