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Overexpression of Krüppel like factor 2 (Klf2) or Klf7 inhibits adipocyte formation. However, it remains unclear whether Klf2 regulates klf7 expression in adipose tissue. In this study, oil red O staining and Western blotting were employed to study the effect of Klf2 overexpression on the differentiation of chicken preadipocytes. The results showed that Klf2 overexpression inhibited the differentiation of chicken preadipocytes induced by oleate and the expression of pparγ, while promoted klf7 expression in chicken preadipocytes. Spearman correlation analysis was used to study the correlation between the expression data of klf2 and klf7 in the adipose tissue of both human and chicken. The results showed that there was a significantly positive correlation between the expression of klf2 and klf7 in adipose tissues (r > 0.1). Luciferase reporter assay showed that overexpression of Klf2 significantly promoted the activity of chicken klf7 promoter (-241/-91, -521/-91, -1 845/-91, -2 286/-91, -1 215/-91; P < 0.05). In addition, the activity of klf7 promoter (-241/-91) reporter in chicken preadipocytes was significantly positively correlated with the amount of klf2 overexpression plasmid transfected (Tau=0.917 66, P=1.074×10-7). Moreover, Klf2 overexpression significantly promoted the mRNA expression of klf7 in chicken preadipocytes (P < 0.05). In conclusion, upregulation of klf7 expression might be one of the pathways that Klf2 inhibits chicken adipocyte differentiation, and the sequence from -241 bp to -91 bp upstream chicken klf7 translation start site might mediate the regulation of Klf2 on klf7 transcription.
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Animals , Humans , Chickens/genetics , Kruppel-Like Transcription Factors/metabolism , Transcription Factors/metabolism , Adipocytes/metabolism , Adipose Tissue/metabolismABSTRACT
Objective@#To explore the effects of Kruppel⁃like factor 7 (KLF7) on the proliferation , migration and cycle of esophageal squamous cell carcinoma cell line Eca109.@*Methods@#Bioinformatics analysis was used to study the expression pattern of KLF7 in esophageal squamous cell carcinoma and its relationship with prognosis and pathological grade. The expression pattern of KLF7 in esophageal squamous cell carcinoma cells was studied by real⁃time PCR , Western blot and cellular immunofluorescence in vitro. The KLF7 overexpression in Eca109 cells was carried out by transfection of the plasmid of pcDNA⁃3. 10⁃KLF7 , and the effects of KLF7 on the proliferation and migration of ESCC cells were detected by CCK⁃8 assay , plate clone formation assay and Transwell assay , and the cell cycle was studied by Flow cytometric analysis.@*Results@#KLF7 was expressed in the tissues and cells of esophageal cancer, and its expression was significantly associated with the pathological grade of esophageal cancer patients ( P <0. 05) , however, there was no significant associaition of KLF7 expression with the survival rate of patients. The expression level of KLF7 in esophageal cancer cells ( Eca109 and EC9706) was significantly higher than that in normal esophageal epithelial cells (Het⁃1A , P < 0. 05) . In addition , the expression of KLF7 was higher in the cytoplasm and nucleus of Het⁃1A cells , while only in the nucleus of esophageal cancer cells (Eca109 and EC9706) Additionally , the overexpression of KLF7 in Eca109 cells changed the cell cycle , promoted cell proliferation and migration , and up⁃regulated the expression levels of CCND1 and P53 (P < 0. 05) .@*Conclusion@#Overexpression of KLF7 might promote the proliferation and migration of esophageal cancer cells.
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Objective @#To explore the effects of the nerve growth factor neutralizing antibody ( anti⁃NGF) and γ⁃secretase inhibitor 3,5⁃difluorophenylacetyl⁃L⁃alanyl⁃S⁃ph ⁃enylglycine⁃t⁃butyl ester (DAPT) on the nuclear expression of p75 neurotrophin receptor (p75NTR ) in esophageal cancer Eca109 cells and on cell proliferation and invasion.@*Methods @#Immunofluorescence cytochemistry was used to detect the expression changes of p75NTR and Ki6 in Eca109 cells before and after treatment with the anti⁃NGF and γ⁃secretase inhibitor DAPT; CCK⁃8 kit and Transwell cell invasion experiment were used to detect the changes of cell proliferation and invasion ability before and after treatment of Eca109 cells with the anti⁃NGF and γ⁃secretase inhibitor DAPT alone or in combination.@*Results@#Immunofluorescence cytochemistry showed that after treatment of cells with the anti⁃NGF and γ⁃secretase inhibitor DAPT, the number of cells expressing p75NTR in the nucleus decreased, and after the combined treatment of cells with the anti⁃NGF and γ⁃secretase inhibitor DAPT, the number of cells expressing p75NTR in the nucleus was the least, and the difference was statistically significant (P < 0. 05); CCK⁃8 method and Transwell cell invasion experiment showed that after treatment of cells with the anti⁃NGF and γ⁃secretase inhibitor DAPT, cell proliferation and invasion ability were correspondingly weaker than those before treatment, and the difference was statistically significant (P < 0. 05) .@*Conclusion @#The anti⁃NGF and γ⁃secretase inhibitor DAPT not only inhibit the nuclear transfer of p75NTR , but also reduce the nuclear expression rate of p75NTR and weaken the cell proliferation and invasion ability accordingly.
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Objective To study and explore Glucose-reducing effect of induced human bone mes-enchymal stem cells on diabetic mice.Methods The diabetic model of rats was duplicated by injection of STZ intraperitoneally.The induced cells were implanted into diabetic mice.Blood glucose levels were moni-tored every 3 days after implantation for 14 days.Results The mice receiving the treated Cells began to decrease their blood glucose levels after 3days.But control Animals that did not receive induced cells exhib-ited persistent hyperglycemia.Conclusions Induced cells by hBMSCs can decrease blood glucose levels on diabetic mice.
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Objective To observe the expression of tumor stem cell markers P 75NTR,Oct-4,Sox-2,Lin28 and Nanog in the tumor sphere from esophageal squamous cells carcinoma Eca 109 and identify the esophageal squamous cell cancer stem cell marker .Methods The serum-free culture method was used for generating tumor spheres: proliferation was observed in enrichment culture tumor spheres .Small tumor spheres were obtained after 5 days culture and big and round tumor spheres appeared after 14 days culture which were collected for experiments and passaged .The expression and location of P75NTR,Oct-4,Sox-2,Lin28, and Nanog were detected by immunofluorescence cytochemistry .Results The expressions of P75NTR,Oct-4 and Lin28 were positive in the center of tumor spheres and some on cytoplasm and other in nuclei of Eca109 monolayer cells.However, Oct-4 fluorescence intensity was weaker than P75NTR.The expressions of Sox-2 and Nanog were positive in cytoplasm of tumor spheres and Eca 109 monolayer cells .Conclusion The cells expressing P75NTR, Oct-4, and Lin28 in the center of the tumor sphere may be esophageal cancer stem cells .
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Spillage of cyst contents during surgical operation is the major cause of recurrence after hydatid cyst surgery. Instillation of a scolicidal agent into a hepatic hydatid cyst is the most commonly employed measure to prevent this complication. SB202190 is a pyridinyl imidazole derivative and is known to be a specific inhibitor of p38 MAPK. In the present study, the scolicidal effect of SB202190 was investigated. Freshly isolated Echinococcus granulosus protoscolices were subjected to SB202190 treatment (10, 20, 40, and 80 microM), and the effects on parasite viability were monitored by trypan blue staining. Corresponding effects were visualized by scanning and transmission electron microscopy. Dose-dependent protoscolex death within a few days of SB202190 treatment was observed. Although the in vitro scolicidal effect of SB202190 was satisfactory, the in vivo efficacy of this drug and also possible side effects remain to be further investigated.
Subject(s)
Animals , Anthelmintics/pharmacology , Dose-Response Relationship, Drug , Echinococcus granulosus/drug effects , Imidazoles/pharmacology , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Parasitic Sensitivity Tests , Pyridines/pharmacology , Survival AnalysisABSTRACT
Objective To study the effects of low dose sodium arsenite to human bone marrow mesenchymal stem cells(hBMSCs) differentiation during establishing of arsenic-resistant cell model. Methods hBMSCs were prepared in conventional method and continuously exposed to 1μmol/L sodium arsenite for ≥12weeks inv vitro. Forty-eight hours acute arsenite toxicity test was drived to assay if the cells acquired arsenic-resistance. The proliferation capacity of CAsE-hBMSCs was observed by the rate of colony formation.The expression of Oct-4 in CAsE-hBMSCs was assayed by RT-PCR and immunocytochemistry. The expression of ABCG2 in CAsE-hBMSCs was analyzed by RT-PCR. Results hBMSCs continuously exposed to 1μmol/L sodium asenite for ≥12 weeks exhibited dramatic resistance to acute arsenite toxicity. The LC 50 for acute arsenite exposure in CAsE-hBMSCs was 35.59μmol/L versus 18.04μmol/L in control cells. Compared to control cells, the CAsE-hBMSCs didn't show malignant proliferation ability. Expression of Oct-4 gene was positive in 4th, 18th passage hBMSCs and the hBMSCs induced by arsenite for 4 weeks but negative in CAsE-hBMSCs. The expression of Oct-4 protein was positive and weakly positive in 4th passage hBMSCs and CAsE-hBMSCs respectively, and the positive granules of Oct-4 distributed in cytoplasm. The expression of ABCG2 gene in CAsE-hBMSCs was obviously lower than that in control cells ( P <0.001). Conclusion Human bone marrow mesenchymal stem cells could be induced to differentiation by low dose sodium arsenite.
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To accomplish the teaching target on clinical practice in Gynecology and Obstetrics for seven-year program,we should consider the features of clinical practice in Gynecology and Obstetrics,choose appropriate teaching methods and make best use of entrance guidance and teaching ward-round and so on to improve students'comprehensive ability.
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Objective To obtain stable arsenic-resistance cells,the fetal bone marrow mesenchymal stem cells(BMSCs)were exposed to low-level arsenite for 18 weeks. Methods Cells from 4 months fetal bone marrow were cultured in ?-MEM medium to obtain BMSCs.After 24h cytotoxicity test of the fetal BMSCs,we chosed 1?mol/L NaAsO_2 to be the dose under which the cell death-rate was 5%-10%.The fetal BMSCs were exposed to low-level arsenite.MTT was used to detect the survival rate and IC_(50) of arsenic-exposed cells and the control cells,which can reflect the change of arsenic tolerance.A hydride generationatomic fluorescence spectrometry method was used to detect arsenic in the arsenic-resistance cells and the control cells.In order to study the mechanism of arsenic-resistance,we also examined the intracellular GSH and GST content in the arsenic-resistance cells and the control cells. Results The fetal BMSCs were continuously exposed to 1?mol/L NaAsO_2.Parallel cells were cultured in medium without arsenic provided passage-matched control.After the fetal BMSCs were continuously exposed to low level NaAsO_2 for 18 weeks,cells exhibited dramatic resistance to acute arsenite toxicity.Compared to control cells,arsenic-resistance cells showed reduction in arsenic accumulation in cells.The GSH levels and GST activity of arsenic-resistance cells were higher than that of control cells.Conclusion Acquisition of stable arsenic-resistance in BMSCs was available by using cell culture and adding low-level arsenic.Our research established a solid basis for further study about the mechanisms of arsenic-resistance in human being.
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Objective To compare the level of interleukin 6 (IL 6),interleukin 8 (IL 8),human chorionic gonadotropin(hCG) with transvaginal ultrasonographic measurement in prediction of the cervix ripening and the time of term labor Methods The 79 cases of primiparous women of term pregnancy were chosen as the research subjects The maternal level of IL 6,IL 8,hCG in cervicovaginal secretions were measured The cervical length, internal cervical os wedge width and forebag length were measured by transvaginal ultrasonography The cervical Bishop score was also determined Results (1) The levels of IL 6,IL 8,hCG in cervicovaginal secretions were significantly higher in women they are in labor than that of women at term not in labor (782?508) ng/L, (10 539?8 680) ng/L,(114?86) IU/L, versus (155?75) ng/L, (7 113?6 050) ng/L,(35?21) IU/L, respectively (2) The levels of cervicovaginal secretions IL 6,IL 8,hCG and the length of cervical, forebag were significant correlation with the cervical Bishop score ( P