2.Construction of pEGFP-N1-TGF-β1 recombinant plasmid and transfected it into primary cultured AEC-Ⅱ of neonatal piglet using lipofectamine 2000
Chinese Journal of Emergency Medicine 2016;25(1):50-56
Objective To construct pEGFP-N1-TGF-β1 recombinant plasmid and transfect it into primary cultured neonatal piglet type Ⅱ alveolar epithelium cell (AEC-Ⅱ) by using lipofectamine 2000,in order to provide basis of methodology for producing recombinant plasmids for transplantation of transfected AEC-Ⅱ into ALI/ARDS animal model lungs.Methods PCR primers were designed to amplify the human TGF-β1 cDNA fragment from plasmid.XhoI and EcoRI were used for double digesting the empty plasmid pEGFP-N1 and cDNA fragment of human TGF-β1.Then the products of double enzyme digestion by using T4 DNA ligase were connected and transformed into DH5α and cultured over night for 16 hours.The structure of recombinant plasmid was identified by using PCR and base sequencing to verify the correctness of pEGFP-N1-TGF-β1 recombinant plasmid.It was then transfected into primarily cultured AEC-Ⅱ by lipofectamine2000 mediated transfection and cultured for another 48 hour.Plasmid DNA (pEGFP-N1-TGF-β1 recombinant plasmid) and lipofectamine 2000 were added into serum-free DMEM respectively,then DNA suspension and Lipofectamine 2000 suspension were blended together and added into cells.After 24-48 hours later,the expression level of enhanced green fluorescent protein (EGFP) was evaluated under fluorescence microscope.Results The structure of vector was verified as pEGFP-N1-TGF-β1 recombinant plasmid by using PCR and base sequencing.Green fluorescence found in some cells showed that the pEGFP-N1-TGF-β1 recombinant plasmids had been successfully transfected into primary cultured AEC-Ⅱ,however,the transfection efficiency still need tobe further improved such as repeating the transfection procedure once again or using adenovirus mediated transfection method to improve the efficiceny and to transplant the cells into animal lungs eventually.Conclusions pEGFP-N1-TGF-β1 recombinant plasmid was successfully constructed and,for the first time,transfected into primarily cultured AEC-Ⅱ of newborn piglets.This established method should be useful for investigation of therapeutic effect and outcomes of lung with experimental acute lung injury.
3.Relationship of hypoxia-inducible factor-1 with the occurrence and development of tumor
International Journal of Surgery 2009;36(7):491-494
Hypoxia-inducible factor-I is the critical factor existed in mammal and human cells which plays an important role in maintaining the oxygen balance. Many hypoxia-responding genes can be activated by H1F-1. So, the relationship between HIF-land matrix matalloproteinases/stromal-derived factor-I/special CXC chemokin receptor/angiogenesis related factors is the hot spot in research of the tumor invasion and me-tastasis.
4.Removal of Excessive Arsenic From Drinking Water by Electric Absorb Technology
Journal of Environment and Health 1992;0(02):-
Objective To control the hazardous effects of excessive arsenic in drinking water on human health. Methods The experiment on the removal of excessive arsenic from drinking water by electric absorb technology (EST) was performed. Results The pilot experiment and experiment in situ showed that, after treatment by EST, the contents of arsenic in source water(0.06-0.33 mg/L) decreased to the levels of
5.The effect of sports training on endothelin-1 and endothelin-1 receptor and its subtypes in the myocardium
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(11):-
Objective To explore the effects of sports training on endothelin-1 ( ET-1 ) and endothelin-1 receptor (ETR) and its subtypes in the myocardium in rats. Methods After sports training, levels of endothelin-1 were measured using radioimmunoassay, and changes in ETAR and ETBR levels by radioligand binding assay. Results The endothelin-1 level in myocardial cells was significantly reduced ( P
6.Galectin-9 and inflammation
International Journal of Pediatrics 2011;38(6):575-578
Galectin-9 is a novel eosinophil chemoattractant,which belong to Galectin family.Galectin-9exhibits a variety of biological functions,such as cell differentiation,maturation,adhesion and aggregation,cell chemotaxis,activity,and cell death.Galectin-9 may play important roles in the development of acute inflammation as well as chronic inflammation associated with allergies,autoimmune diseases,infectious processes.This review summarizes the advances in Galectin-9 study.
7.Diagnostic significance of procalcitonin for postoperative infections in patients with bone fractures
Chinese Journal of Clinical Infectious Diseases 2009;2(3):159-161
Objective To investigate the significance of procalcitonin(PCT),C-reactive protein (CRP)and WBC counts in predicting the postoperative jnfections in patients with bone fractures.Methods Clinical data of 49 patients with open fractures were retrospectively analyzed,and the patients were grouped into the infeeted(n=24)and non.infected(n=25).PCT,CRP and WBC values were detected 1,4,7and 10 days after the operations,and the results were compared with those taken from the healthy controls.Areas under the receiver operating characteristic(ROC)curves were used to determine the value in predicting postoperative infections.Results PCT,CRP and WBC values in the infected group and the noninfected group were higher than those in health controls(F=19.84,57.71 and 35.44,P<0.01).PCT,CRP and WBC values in the infected group were higher than those in the non-infected group 4.7 and 10 days after the operations(tPCT=7.31,7.74 and 4.59;tCRP=2.23,5.75 and 8.01;tWBC=2.34,2.51 and 4.07,P<0.05).ROC curves suggested that PCT value was more sensitive and had higher specificity than CRP and WBC values in the diagnosis of postoperative infections.Conclusions PCT,CRP and WBC values arise in bone fracture patients with postoperative infections.Detection of PCT has higher sensitivity and specificity.which can be used in early diagnosis of postoperative infections.
9.History and current status of cervical intervertebral disc replacement
Chinese Journal of Tissue Engineering Research 2007;0(35):-
Cervical intervertebral disc replacement effectively tackles limited motion range of the neck and long-term adjacent degeneration acceleration following traditional anterior cervical fusion and fixation. This paper reviews history of cervical intervertebral disc replacement,current prosthesis types,related indications and complications,and further investigates the feasibility of minimally invasive replacement.