1.Expression of adenovirus-mediated CTLA4Ig-IRES-CTLA4 gene in bone marrow mesenchymal stem cells
Haiying LUO ; Yunfang WANG ; Xue NAN
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
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.
2.The teaching reform and discipline construction in medical function
Ying AN ; Nan SHEN ; Lijing ZHAO ; Xue CHEN
Chinese Journal of Medical Education Research 2002;0(01):-
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.
3.Assessment and treatment of halitosis.
Chinese Journal of Stomatology 2013;48(10):627-631
Anti-Infective Agents, Local
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therapeutic use
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Biosensing Techniques
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Chlorhexidine
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therapeutic use
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Chlorine Compounds
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therapeutic use
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Chromatography, Gas
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Dehydroascorbic Acid
;
therapeutic use
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Dental Disinfectants
;
therapeutic use
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Halitosis
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diagnosis
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therapy
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Humans
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Hydrogen Peroxide
;
therapeutic use
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Mouthwashes
;
therapeutic use
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Odorants
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prevention & control
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Oils, Volatile
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therapeutic use
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Oral Hygiene
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instrumentation
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Oxides
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therapeutic use
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Sodium Bicarbonate
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therapeutic use
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Sulfur Compounds
;
analysis
4.The mechanism and influencing factors of halitosis.
Chinese Journal of Stomatology 2013;48(9):566-569
5.Two special cerebral hyponatremia.
Xiang-zhen NAN ; Yong-qing WANG ; Chun-xue YAN
Chinese Journal of Pediatrics 2003;41(9):715-717
6.Cytotoxicity of a new type of antibiotic stainless micro-screw implant
Dan ZHANG ; Yang ZHANG ; Li LU ; Ke YANG ; Nan XUE
Chinese Journal of Tissue Engineering Research 2010;14(16):2916-2920
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.
8. “Spectral toxicity” relationship of acute toxicity of Sophora alopecuroides extracted by different extraction methods to mice
Chinese Traditional and Herbal Drugs 2019;50(11):2656-2664
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.
9.Function and structure analysis of premembrane and envelope proteins of Zika virus
Chang LU ; Na LI ; Pu XUE ; Nan LI ; Ping CHEN
Chinese Journal of Zoonoses 2017;33(1):81-84
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.
10.Effect of coenzyme Q10 on the expression of interleukin-17 and interleukin-23 in gingival tissue of diabetic rats with periodontitis
Yanli XU ; Yi XUE ; Zhongyin WU ; Nan ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(5):707-711
BACKGROUND:Coenzyme Q10 participates in the electron transport of respiratory chain and possesses antioxidant, anti-apoptotic and anti-inflammatory properties. It has achieved good outcomes in cardiovascular disease, diabetes and cancer. Coenzyme Q10 may also have a certain application value in the fields of diabetes and periodontitis. OBJECTIVE:To observe the effect of coenzyme Q10 on the expression of interleukin-17 and interleukin-23 in gingival tissue of type 2 diabetic rats with periodontitis.METHODS:Forty-eight healthy male Wistar rats were randomly divided into 3 groups: control, periodontitis+ diabetes+physiological saline and periodontitis+diabetes+coenzyme Q10. Rats in the control group were fed with normal diet and water. Rats in the periodontitis+diabetes+physiological saline and periodontitis+diabetes+ coenzyme Q10 groups were subjected to induction of periodontitis using the method of silk ligation and type 2 diabetes by feeding a high-fat and high-sugar diet and intraperitoneal injection of streptozotocin. After successful modeling, rats in the periodontitis+diabetes+coenzyme Q10 group were intragastricaly administered coenzyme Q10 for 8 successive weeks. Rats in the periodontitis+diabetes+physiological saline group were administered equal amount of physiological saline. At the end of 2nd, 4th and 8th weeks after drug administration, four rats were randomly selected and sacrificed. The expression levels of interleukin-17 and interleukin-23 in gingival tissue were detected by immunohistochemistry. RESULTS AND CONCLUSION: At the end of 8th week, interleukin-17- and interleukin-23-positive expression in the periodontitis+diabetes+physiological saline group was significantly higher than that in the periodontitis+ diabetes+coenzyme Q10 group (P < 0.05). Coenzyme Q10 can reduce the expression levels of interleukin-17 and interleukin-23 in gingival tissue of type 2 diabetic rats with periodontitis, and aleviate periodontal tissue inflammation of type 2 diabetic rats with periodontitis.