Objective To construct adenovirus vector harboring CTLA4Ig-IRES-CTLA4 gene, and to express the gene in bone marrow mesenchymal stem cells (BMMSCs). Methods The cDNA fragments of CTLA4, and the extracellular domains of CTLA4 and IgGFc were cloned to pT-Easy vectors by RT-PCR from the activated splenocytes of Wistar rats. Recombinant adenovirus vector harboring CTLA4Ig-IRES-CTLA4 was produced from AdeasyTM Adenoviral Vector System by homologous recombination between the Adtrack vector (pAdtrack-GFP-CTLA4Ig-IRES-CTLA4) and the plasmid pAdEasy-1 containing most of the human adenovirus serotype 5 (Ad5) genome in E. coli BJ5183 strain, and then packaged and propagated in 293 cells. BMMSCs were infected with the recombinant adenovirus, and the co-expression of both CTLA4Ig and CTLA4 was then detected by RT-PCR and Western blotting. The immunosuppression function of the transfected BMMSCs was investigated by mixed lymphocyte response (MLR). Results Fusion gene CTLA4Ig was constructed successfully, and the recombinant CTLA4Ig-IRES-CTLA4 adenovirus was then generated by homologous recombination and packaged in 293 cells. The transduction efficiency of recombinant adenovirus in BMMSCs was elevated up to 98.67% determined by flow cytometry using a GFP as marker. CTLA4Ig and CTLA4 were expressed at the same time in BMMSCs detected by RT-PCR and Western blot, and the transfected stem cells showed stronger immunosuppression function than that of BMMSCs or BMMSCs transduced by Ad-CTLA4Ig. Conclusion The BMMSCs transfected by the recombinant adenovirus can co-express CTLA4Ig and CTLA4, which improved the immunosuppression function of BMMSCs in the way of costimulatory pathway blockade.

In constructing the system of medical functional teaching and enhancing the quality and ability of students,we adjust the content and method of teaching in practice,and try to find students'problems in study through questionnaire survey and promote teaching reform and disciplines construction.

Alkaloids ; Antineoplastic Agents, Phytogenic ; Chemical Phenomena ; Chemistry ; Harringtonines

Anti-Infective Agents, Local ; therapeutic use ; Biosensing Techniques ; Chlorhexidine ; therapeutic use ; Chlorine Compounds ; therapeutic use ; Chromatography, Gas ; Dehydroascorbic Acid ; therapeutic use ; Dental Disinfectants ; therapeutic use ; Halitosis ; diagnosis ; therapy ; Humans ; Hydrogen Peroxide ; therapeutic use ; Mouthwashes ; therapeutic use ; Odorants ; prevention & control ; Oils, Volatile ; therapeutic use ; Oral Hygiene ; instrumentation ; Oxides ; therapeutic use ; Sodium Bicarbonate ; therapeutic use ; Sulfur Compounds ; analysis

Halitosis ; etiology ; metabolism ; microbiology ; Helicobacter Infections ; complications ; Helicobacter pylori ; Humans ; Oral Hygiene ; Periodontal Diseases ; complications ; Smoking ; Volatile Organic Compounds ; metabolism

Diagnosis, Differential ; Humans ; Hyponatremia ; diagnosis ; etiology ; therapy ; Inappropriate ADH Syndrome ; diagnosis ; etiology ; therapy

Objective To investigate the effects of different doses of propofor on ATP-sensitive K~+currents(I_KATP)in human atrial myocytes and the underlying mechsnism.Methods A small piece of myocardiumwas obtained from right atrium in patients undergoing atrial septal defect or ventricular septal defect surgery.Themyocardium specimen was placed in cold Ca~(2+)-free cardioplegic solution aerated with 100% oxygen.Themyocardium specimen was cut into small chunks(1 mm~3).Atrial myocytes were isolated by enzymatic dissociationtechnique.The effects of propofol on I_KATP in atrial myocytes were studied using the whole-cell configuration ofpatch-clamp technique.Results The outward currents were recorded with a pipitte solution containing 0.3mmol?L~(-1) ATP.The currents were inhibited by glibendamide 10 ?mol?L~(-1),a specific K_ATP channel inhibitor,suggesting that the outward currents were I_KATP.I_KATP aws activited by propofol in a dose-dependent manner.Conclusion Propefol can activate the I_KATP in human myocytes in a concentration-dependent manner and themechanism of its myocardial depressant action may be partly explained.

Objective: To establish the chemical fingerprint of Sophora alopecuroides extracts based on high performance liquid chromatography (HPLC), and determine the LD50 of different extracts of S. alopecuroides to analyze its “spectrum toxicity” relationship. Methods: A series of extracts were prepared by 75% ethanol reflux (ER), water decoction (WD), 75% ethanol ultrasound (EU) and water ultrasound (WU), and their fingerprints were established to determine the acute toxicity LD50 of different extracts. The relationship between chemical composition and acute toxicity LD50 of S. alopecuroides extracts were studied by means of fingerprint similarity evaluation system. Results: The LD50 of ER, WD, EU, and WU extracts were 38.397, 24.994, 18.536, and 19.957 g/kg, respectively. The ocular lesions of mice viscera were mainly manifested in liver and kidney, and the toxicity of ER extracts was the greatest. The 10 common peaks of S. alopecuroides extracts can be divided into two categories; Peaks 4 and 10, oxymatrine and sophocarpidine were negatively correlated with acute toxicity LD50. Conclusion: The spectral toxicity relationship analysis method of S. alopecuroides was constructed. The unidentified peaks 4, 10 and oxymatrine and sophocarpidine were the main chemical components of the toxicity reaction, which laid a good foundation for clinical application and scientific and rational development of S. alopecuroides.

BACKGROUND: Prevention of implant from inflammation was an effective method to reduce expulsion rate of stainless steel micro-screw implant, and develop new type of antibiotic material.OBJECTIVE: To evaluate the cytotoxicity of a new type of antibiotic stainless steels.METHODS: Metal test samples (antibiotic stainless steel, medical stainless steel, and medial pure titanium) were made into rectangular solids with length of 15 mm × 10 mm × 3 mm. Samples were cleaned with high temperature and high pressure. Alloy leaching liquor was prepared with DMEM culture media according to the ratio betwean surface area and volume of culture solution (3 cm~2/mL). The leaching liquor was maintained in incubator at 37 ℃ for 96 hours, and then degerming was performed using microporous membrane. 6.4% phenol was added, which was considered as the positive control group, and DMEM culture media was considered as the blank control group. Growth of MG63 cells was observed under inverted phase contrast microscope;absorbanca of cells cultured for 24, 48, 72, 96, and 120 hours was detected using MTT test; cytotoxicity of antibiotic stainless steels was evaluated.RESULTS AND CONCLUSION: ① At 24 hours after culture, calls in the positive control group was abnormal; while, cells in other groups were well adherent-grew. ② After 48 hours of culture, with the culture time increased,cytotoxicity was detected out in the positive control group; cells in other groups and blank control groups were normal and grew well. Afew of cells in stainless steels group showed karyopyknosis. ③ The absorbance was the highest of medical pure titanium, and then of antibiotic stainless steel and of medical stainless steel, while there was no significant difference between the three materials. ④ The level of cytotoxicity was grade 0. The results suggested that the antibiotic stainless steel which had the same cytotoxicity grade as medical stainless steel and pure titanium was in line with the requirement of its clinical application.

The premembrane and envelope proteins (prM-E),which contains the mainly protective antigen related with virulence and tropism,are the primary structural protein of flavivirus.However,prM-E in ZIKV is rarely understood.We have analyzed the structure and biological effects of prM-E in ZIKV by bionformatics methods.The prM-E proteins virus-like particles of dengue virus was introduced in the present.Then,the prM-E proteins virus like particles of ZIKV was prospected.