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Objective:To investigate the effect of piggyBac transpon,as a carrier of four defined transcription factors Oct4,Sox2,Klf4 and c-Myc,in the reprogramming of mouse embryonic fibroblasts (MEFs)to induced pluripotent stem cells (iPSCs).Methods:The MEFs were isolated from Oct4-GFP fetal mice and transfected by piggyBac transposon with four factors (Oct4,Sox2,Klf4 and c-Myc).The morphological changes of clones were traced with microscope during the process of induction.The chromosomes were analyzed to evaluate the karyotypic variation of iPSCs.The mRNA expressions of Oct4, Nanog and FGF4 associated with embryonic stem cells (ESCs)in the iPSCs of mice were tested by RT-PCR;the protein expressions of SSEA-1,Nanog and Alkaline phosphatase in the iPSCs of mice were determined by flow cytometry,immunofluorescence and AP staining.The iPSCs were transplanted into the NOD-SCID mouse groin,4 weeks later,the teratomas were removed for HE staining and the differentiation of tissue was observed.Results:The iPSCs were successfully obtained from MEFs by piggyBac carrying Oct4,Sox2,Klf4,and c-Myc.The round or oval iPSCs clones were similar to ESCs with clear boundry and large dense nuleus.The iPSCs showed the normal karyotypic and expressed the marker genes (Oct4,Nanog and FGF4)and proteins (SSEA-1,Nanog and AP)of ESCs.Teratomas containing three germ layers were formed in NOD-SCID mice after tanspalantation of iPSCs.Conclusion:The iPSCs are reprogrammed from MEFs by piggyBac transposon with four transcription factors-Oct4,Sox2,Klf4 and c-Myc,and the iPSCs with normal karyotype possess the characteristics of ESCs.
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Objective To study effects of oligo-peptide I-C-F-6 in carapax trionycis on rats with liver fibrosis induced by CCl4;To discuss its anti-liver fibrosis effects and possible mechanisms. Methods Forty-eight SD male rats were randomly divided into normal control group, model group, bifendate group, and oligo-peptide I-C-F-6 group, 12 in each group.CCl4 was injected intraperitoneally to build rat liver fibrosis model.Oligo-peptide I-C-F-6 group and bifendate group were given subcutaneous injection of oligo-peptide I-C-F-6 (0.12μg/g) or bifendate (0.12μg/g). At the same time, normal control group and model group were giventhe same volume of saline for seven weeks. The levels ofALT, AST,MDA, SOD, IL-4, IL-10 and TNF-α were tested.The histomorphology changes were observed under optical microscopeby HE, and the expressions of transforming growth TGF-β1 were determined by immunohistochemistry.Results Compared with model group, serum levels of ALT and AST were reduced evidently in oligo-peptide I-C-F-6 group. Hepatic content of MDA, IL-4 and TNF-α decreased, while SOD activity and IL-10 were found significantly increased. Liver fibrosis was ameliorated significantly. Hepatic expressions of TGF-β1 were weakly positive.Conclusion Oligo-peptide I-C-F-6 can ameliorate hepatocyte damage of model rats, thus it has anti-oxidative and anti-liver fibrosis effects on liver fibrosis in rats.
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[ ABSTRACT] Diabetes is characterized by an absolute or relative deficiency inβ-cell mass, which cannot be re-versed with existing therapeutic strategies.The restoration of the endogenous islet β-cells can stabilize the level of blood glucose.The isletβ-cells can be obtained from the directional differentiation of stem cells, but the process is complex and has the risk of teratomas generation.Cell direct reprogramming, one terminal differentiated cell can transdifferentiate into another kind of terminal differentiated cell, which is other than directional differentiation from stem cells.Direct reprogram-ming gives rise to the generation of isletβ-cells from one terminal differentiated cell, may be preferable for diabetes therapy because of its unique advantage.
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@#Objective To explore the effect of the dexamethasone on the levels of cytokine in the brain of endotoxin shock rats. Methods 54 Sprague-Dawley rats were randomized to the sham group, LPS group (lipopolysaccharide 8 mg/kg, i.v.) and DEX group (dexamethasone 5 mg/kg, i.v., in addition). The blood pressure was measured dynamically. The contents of interleukin (IL)-4, IL-10 and tumor necrosis factor (TNF)-α were detected with ELISA 1 h, 3 h, 6 h after shock. Results As the occurrence and development of shock, blood pressure went down gradually, IL-4 and IL-10 decreased (P<0.01), and TNF-α increased (P<0.01) in LPS group, while the TNF-α decreased (P<0.01), the IL-4 and IL-10 increased (P<0.01) in the DEX group compared with the LPS group. Conclusion Dexamethasone may protect the brain from the endotoxin shock injury in rats through regulating the inflammatory factor.
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Objective:To observe the changes of glyeemia, urine protein, serum creatinine, blood urea nitrogen and glycosylated hemoglobin in diabetic nephropathy rats and treating effects of Xiaokekang. Methods: Streptozotocin (STZ) was abdominally administered and was feeded by high lipid diet to establish diabetic nephropathy model in rats. Animals were divided into three groups: model group, Xiaokekang treatment group and normal group. Changes in glycemia, serum lipids, serum creatinine, blood urea nitrogen were measured at the 8th, 12th and 16th week after STZ injection. Glycosylated hemoglobin was measured at the 16th weeks after STZ injection. Results: Glycemia, serum lipids, serum creatinine were higher at the 8th, 12th and 16th week compared with normal group. Serum creatinine was higher at the 12th and 16th week, and glycosylated hemoglobin was also highter than that of the normal group. Xiaokekang reduced these changes. Conclusion: Xiaokekang can reduce glycemia, serum lipids and protect renal function.
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Objective: To investigate selected cardiovascular responses of post myocardial infarction patients(PMIP)during exercise within the context of different levels of ST segment depression. Method: Forty-six male recent PMIP performed a graded exercise test on a motorized treadmill during which time blood pressure and heart rate were measured and a 12 lead electrocardiogram (ECG) was monitored continuously. They were then subdivided into those ST segment depression ＜1.0 mm at exercise stage Ⅱ (group 1) and those who had ST segment depression ＞1.0 mm at this stage (group 2). Result: The results showed that the patients with a larger degree of ST segment depression at the given work load exhibited higher a value of rate-pressure product (RPP).Conclusion: RPP may be of particular value in estimating the clinical response to exercise when ECG monitoring is not available.
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AIM: To explore the mechanism of Nao-re-qing oral liquid (NRQ) decreasing endotoxin (ET)-induced fever in rabbits. METHODS: (1) The ET-induced fever model was established in rabbits. Febrile response of rabbits was observed. (2) The arginine vasopressin (AVP) content in the ventral septal area (VSA),and cAMP content in hypothalarmus (HP) and CSF were determined by radioimmunoassay.RESULTS: (1) In ET group,the maximal increment in body temperature (?T) [(1.80?0.16) ℃],6 h thermal respone index (TRI_6)(11.31?0.20),the cAMP content in the HP [(1.35?0.21)nmol/g],the cAMP content in CSF [(66.69?1.82) nmol/L] and AVP content in the VSA [(30.80?9.59)ng/g ] were significantly higher than those in NRQ+ET group[?T(0.82?0.08) ℃,TRI_6(5.73?0.09),HP: cAMP(0.70?0.50)nmol/g,CSF: cAMP(56.86?1.34),AVP:(11.91?3.47)ng/g]( P
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AIM: To investigate the antipyretic mechanism of Naoreqing (NRQ) oral liquid, a Chinese medicine. METHODS: ① Fever models of rabbits were established by intravenous. injection of endogenous pyrogen (EP). ② The antipyretic action of NRQ were observed. ③ cAMP contents in the hypothalamus (HP) and the cerebrospinal fluid (CSF) were determined by radioimmunoassay. RESULTS: ① NRQ obviously reduced body temperature of febrile rabbits induced by EP (P