2.Construction of human telomerase reverse transcriptase-immortalized rat bone marrow mesenchemal stem cell strains
Huiping LIU ; Xiaolong ZHONG ; Qing ZHAO ; Wenqi HUANG ; Ke AN
Chinese Journal of Tissue Engineering Research 2015;(23):3621-3627
BACKGROUND:Because of convenient source, multi-lineage differentiation and low immunogenicity, bone marrow mesenchymal stem cels are the ideal cel type to serve as vectors of transgenic cels in pain management. However, the replicative senescence and smal amount of cels obtained from the bone marrow restrict the application of bone marrow mesenchymal stem cels in pain research. OBJECTIVE:To construct human telomerase reverse transcriptase (hTERT)-immortalized rat bone marrow mesenchymal stem cels as transgenic celular vectors for pain therapy. METHODS:Bone marrow mesenchymal stem cels were obtained from whole rat bone marrow, and then transfected with a lentivirus containing the hTERT (pLV-Puro-EF1α-hTERT) folowed by puromycin selection. hTERT expression and telomerase activity in these transfected cels were determined by RT-PCR and TRAP. Morphological changes, capacity of cel growth and multi-lineage differentiation, chromosome karyotype and tumorigenicity were observed in vitro. Moreover, the expression of cel surface molecule, Nestin, MHC-I and MHC-II in transfected cels were also detected by flow cytometry and immunocytochemistry. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cels geneticaly modified by hTERT could be cultured and passaged through 30 generations in vitro. Compared to the primary and negative transfected cels, the hTERT-modified bone marrow mesenchymal stem cels showed higher expression of hTERT mRNA, telomerase activity and cel proliferation. Most of transfected cels stayed at G2/M and S stages. The proliferation index of the transfected cels were increased dramaticaly. The positive rates of CD29, CD44 and CD90 were over 70%, but the positive rates of CD34 and CD45 were less than 5%. Transfected cels were positive for Nestin in the cytoplasm, but negative for MHC-1 and MHC-11. In addition, this cel line continued to exhibit the characteristics of fibroblastic bone marrow mesenchymal stem cels, including phenotype, differentiation into osteoblasts, adipocytes and neuron-like cels. No chromosome abnormality and tumor formation were observed in this experiment. Taken together, these data suggests that the rat bone marrow mesenchymal stem cels immortalized by hTERT gene are constructed successfuly and stil maintain major stem cels characteristics, which provide safe and stable cel vectors as research base for pain therapy.
3.The clinical value of detecting the interleukin-6 levels in serum in the patients with sepsis
Youcheng ZHAO ; Qiong NAN ; Qing KE ; Hongjian WEI ; Yong DUAN
Journal of Chinese Physician 2001;0(05):-
Objective To investigate the regularity and significance of serum interleukin-6(IL-6) levels in the patients with sepsis.Methods Forty sepsis patients with APACHEⅡ scores over 12 and 30 normal controls were involved in the study.The IL-6 levels in serum were detected with radio-immunity analysis assay and the correlations among the IL-6 levels,the prognosis and APACHEⅡ score were investigated.Results In the patients with sepsis,the average level of IL-6 in serum was(152.02?55.77) pg/ml,which was significantly higher than that of the control group [(85.79?30.96) pg/ml](P
4.CT Diagnosis of Cerebral Infarction After Subarachnoid Hemorrhage
Ke QING ; Zhenguo ZHAO ; Shaobing DENG ; Zebin LUO
Journal of Practical Radiology 2001;0(07):-
Objective To study the CT diagnosis of cerebral infarction after subarachnoid hemorrhage.Methods Clinical and CT findings of cerebral infarction after subarachnoid hemorrhage were analyzed 16 cases.Results Cerebral infarctions occurred in 7~13 d(average 10.5 d)after subarachnod hemorrhage.Multiple and bilaterial low densities in basal ganglia or/and brain lobes were characteristic of cerebral infarction after subarachnoid hemorrhage.Conclusion CT is an effect method for evaluation and diagnosis of cerebral infarction after subarachnoid hemorrhage.It is important to prevent its occurrence because it have high dead rate and bad prognosis.
5.Effect of Bay K 8644 on arteriole smooth muscle cell membrane potential in rats with severe hemorrhagic shock.
Journal of Southern Medical University 2006;26(4):421-424
OBJECTIVETo investigate the effect of Ca(2+) influx through L-type Ca(2+) channels on normal and hyperpolarized membrane potential of arteriole smooth muscle cells (ASMCs) in rats.
METHODSThe ASMCs isolated from normal rats and those with severe hemorrhagic shock were labeled with DiBAC4 (3) for membrane potential detection.
RESULTSCa(2+) influx caused hyperpolarization of the membrane potential in the normal ASMCs but depolarization in the cells from rats with hemorrhagic shock, and this effect could be inhibited by TEA.
CONCLUSIONCa(2+)-activated potassium channels activated by Ca(2+) influx through L-type Ca(2+) channels in normal ASMCs to cause hyperpolarization but leads directly to membrane potential depolarization in ASMCs from rats with severe hemorrhagic shock. This finding can be meaningful for treatment of vascular hyporeactivity in advanced stage of severe shock.
3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester ; pharmacology ; Animals ; Arterioles ; physiopathology ; Calcium Channel Agonists ; pharmacology ; Calcium Channels, L-Type ; metabolism ; Female ; Male ; Membrane Potentials ; drug effects ; Muscle, Smooth, Vascular ; physiopathology ; Potassium Channels, Calcium-Activated ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; physiopathology
7.Clinical analysis on 175 cases of occupational brucellosis.
Yi-wen JIANG ; Qing WANG ; Ruo-xin ZHAO ; Shu-ke GE ; Xin-wei GUO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(11):861-863
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diagnosis
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therapy
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8.AnaIysis of choroid radiographic characteristics of diabetics
Ke-Jun, LI ; Xiao-Bin, ZHAO ; Qing-Min, MA ; Zhi-Yang, JIA
International Eye Science 2015;(3):478-481
· AlM: To observe the characteristic of choroidal circulation in diabetics and investigate its changes as well as the relationship between it and the development and progression of diabetic retinopathy ( DR) .
·METHODS:All 45 diabetics were divided into 3 groups:no diabetic retinopathy ( NDR), nonproliferative diabetic retinopathy ( NPDR ) , proliferative diabetic retinopathy ( PDR);and 20 health people were selected to be control group.All subjects were examined by FFA and indocyaine green angiography ( lCGA ) ( Heidelberg retina tomography, Germany ) at the same time. The characteristics of angiograph results were comparatively observed and the feature of diabetic choroidapathy were analyzed.
· RESULTS: ( 1 ) There were no significant differences between DR groups and control group in the central retinal artery ( CRA ) filling time.There were significant decreases of the choroidal artery filling time in DR groups, compared to the control group (P<0.05).(2) ln all DR groups, the more serious DR, the bigger proportion of reverse filling.( 3 ) With more serious DR, the ratio increased gradually in early phase of choroidal filling deficiency angiograph and in the late phase of spot hyperfluorescence.
·CONCLUSlON:lCGA may be a useful adjunct to FFA in the evaluation of choroidal vascular changes in DR.The research provides that the diabetic choroidal circulation was abnormal before the occurrence of DR, which fully proved the presence of diabetic choroidopathy.
9.The strength of the nodes of the brain white matter weighted networks and its relationship with the course of disease in male schizophrenia
Jianhuai CHEN ; Zhijian YAO ; Jiaolong QIN ; Ke ZHAO ; Rui YAN ; Lingling HUA ; Qing LU
Chinese Journal of Nervous and Mental Diseases 2014;(12):721-725
Objective To explore the differences in the strength of the nodes in the brain white matter weighted networks between the male patients with paranoid schizophrenia and male healthy controls, and to analysis the integrity of the white matter fiber tracts that connected to the different brain regions and its relationship with the course of disease. Methods Diffusion tensor imaging (DTI) data were obtained from 25 male patients with paranoid schizophrenia and 26 male healthy controls. The whole brain was parcellated into 90 regions by using the anatomical label map. Tractography was performed in the whole brain of each subject to reconstruct white matter tracts using the FACT algorithm. The brain white matter weighted networks were then constructed using the complex network theory. Results The strength of the nodes in the networks of schizophrenia significantly decreased in the right thalamus (P=0.03, corrected) and the right hes?chl gyrus (P=0.04, corrected). Negative correlation was found between the strength of the right thalamus and the course of disease (r=-0.45, P=0.03). Conclusion The integrity of the white matter fiber tracts connected to the thalamus and tem?poral lobes in the male paranoid schizophrenia is impaired. The lesion of fiber tracts connected to the thalamus is related with the course of disease.