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Objective To observe the change of JAK-STAT-SOCS signal molecules after interferons alpha acting on the cancerous oral cells in 3 different degrees,namely NOK,DOK and KB cells,and to provide research foundation for the deep understanding of OSCC (oral squamous cell cancer) tumor ceils immune escape mechanism.Methods NOK,DOK,and KB cells were all cultured respectively,and then the third passage cells in the logarithmic growth phase were inoculated in cell culture plate.Blank control group of each hole was added 2 ml complete medium containing 10 ％ FPS.DMSO control group of each hole was added 2 ml complete medium containing 0.1％ DMSO.And in experimental groups containing 10 U/ml,100 U/ml,and 500 U/ml interferons,complete culture medium were added to each hole containing different concentrations of interferons alpha.CP-690550 (100 μmol/L) was added before interferons alpha was added 1 h.All were detected by RT-PCR test and Western blot test after conventional cultured for 24 h.Results RT-PCR detection showed that JAK1 and JAK2 in NOK cells had a small amount of expression,interferons alpha and CP-690550 cells could not influence the expression of JAK1 and JAK2 of NOK group,and there was no statistically significant difference (P ＞ 0.05).Interferons alpha in 100 and 500 U/ml could stimulate the increase of JAK1 and JAK2 expression in DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).CP-690550 could effectively reduce the JAK1 expression of DOK and KB cells,while had no effect on the expression of JAK2,and the differences were statistically significant (P ＜ 0.05).Western blot showed that STAT1,STAT3 and pSTAT3 (Tyr705) all expressed in the control group,while pSTAT1 (Tyr701) didn't express in the control group.Interferons alpha and CP-690550 cells had no effect on STAT1,STAT3 and pSTAT3 (Tyr705) expression of NOK group,and there was no statistically significant difference (P ＞ 0.05).100 U/ml and 500 U/ml of interferons alpha could stimulate the increases of pSTAT3 (Tyr705) expression of DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).While they had no effect on pSTAT1 (Tyr701) expression.CP-690550 could effectively reduce the pSTAT3 DOK and KB cells (Tyr705) expression,and the differences were statistically significant (P ＜ 0.05).Western blot showed that there were expression of SOCS1 and SOCS3 in control group.Interferons alpha and CP-690550 had no effect on SOCS1 and SOCS3 expression of NOK cell group,and there was no statistically significant difference (P ＞ 0.05).100 U/ml and 500 U/ml of interferons alpha could stimulate the increase of SOCS 1 expression of DOK and KB cells,and differences were statistically significant (P ＜ 0.05).For the expression of SOCS3,no influence.CP-690550 could effectively reduce the expression of SOCS1 of DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).Conclusions Interferons alpha activate DOK JAK1 and KB cells and the expression of JAK2,mainly JAK1 activation.Interferons alpha,by activating DOK JAK1 and KB cells and the expression of JAK2,promote STAT3 phosphorylation in Tyr705 locus.Interferons alpha,by promoting STAT3 phosphorylation,further promote the expression of SOCS1,which plays the role in inhibiting interferons alpha and reducing the apoptosis.
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Insulin resistance model of 3T3-L1 adipocytes were prepared with plamotic acid.Adipocytes with generated insulin resistance were cultured with different concentrations of globular domain of adiponectin(gAd:250,500,1 000 ng/ml).The cell culture medium glucose content was detected with the glucose oxidase method,the mRNA expressions of insulin receptor substrate-1 (IRS-1),phosphatidylinositol-3 kinase(PI3K),and protein kinase B(PKB) were detected with real-time quantitative PCR method.The phosphorylation of IRS-1 was detected by Western blot.Compared with the control group,the experimental group showed significantly increased glucose consumption (P ＜ 0.01),and with the increasing gAd concentration,glucose consumption was gradually increasing.IRS-1 phosphorylation was increased gradually with the increasing concentration of gAd.These results suggest that gAd can promote glucose uptake by 3T3-L1 adipocyte model with generated insulin resistance.This may be correlated with promoting insulin signal transduction and improving insulin resistance in adipocytes.
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Objective To explore the expression level of thioredoxin (Trx) in acute myeloid leukemia (AML) cells,and its association with clinical features and prognosis,as well as the effects of diamide,an inhibitor of Trx,in inducing AML cells apoptosis.Methods Expression of Trx on AML cells and fresh bone marrow cells from healthy adults were analyzed by Western-blotting. The inhibition of proliferation was measured by MTT assay.The anti-leukemia effect of diamide was observed by morphology and agarose gel electrophoresis.Results 75 ％ (15/20) of AML patients expressed Trx,while no expression was observed in control group.Higher expression level of Trx was associated with higher WBC counts (x2 =9.375,P ＜ 0.05),which suggested that overexpression was associated with leukemogenesis.The inhibition of diamide on AML cells showed time and dose dependent by MTT assay.The IC50 values of diamide at 24 h,48 h and 72 h were 98.26 mg/ml,47.53 mg/ml and 8.34 mg/ml,respectively.After AML cells were treated with diamide,the apoptotic body appeared by morphology,and the typical DNA “ladder” bands were confirmed by agarose gel electrophoresis.Conclusion Trx expression level in AML cells is significantly higher than that of control group.Diamide inhibites the proliferation of AML cells by inducing apoptosis,which might be a potential agent for AML.
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Objective To study the signal transduction mechanism of nicotine induced periodontal ligament fibroblasts (PDLFs) apoptosis. Methods This study used 1 μg/ml, 10μg/ml and 100μg/ml nicotine to intervene PDLFs cells for 24h separately. NF-κB, p53, I-κB and Caspase3 expression were detected. Results After Nicotine was done on PDLFs cells for 24h, the transcription of p53, and Caspase3,and the translation of Caspase 3 protein were increased, while NF-κB was decreased. At the same time, the transcription of NF-κB decreased gradually with the concentration of nicotine increased ( r = 0. 707, F =33. 705, P ＜0. 01 ), nevertheless, I-κB was reversed ( r =0. 964, F =374. 883, P ＜0. 01 ). p53 expression was increased gradually with the concentration of nicotine increased ( r =0. 957, F = 153. 377, P ＜0. 01).Both Caspase3 mRNA (r =0.935, F =318.371, P ＜0.01) and protein (r =0.677, F =8. 459, P ＜ 0. 05 )increased gradually. Conclusion Nicotine induced PDLFs apoptosis was mediated through NF-κB and p53 pathway.
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Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.
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Objective:To master the information of the communication between undergraduate students and parents,so as to provide basis for strengthening university student's psychology work.Methods:We used the investigation questionnaire designed by ourselves,investigated 841 undergraduate students in ShanXi Medical University and established SPSS 11.5 database carrying on statistical analysis.Results:62.2% students ring up their parents once a week,5% students call once a day,22.7% students call once a month,and 10.1% students wait for their parents to call them.8.4% students think that they have generation gap of communication with their parents,10.3% students fully obey their parents,81.3% students use parental words to encourage themselves.Conclusion:Family is an important factor for students' psychological development.
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AIM: To study the effects of [Gly~(14)]-humanin,one of the strongest derivate of humanin,on proliferation and differentiation of neural stem cells(NSCs) and the protective action of cell death or apoptosis induced by ?-amyloid protein 1-42.METHODS: NSCs were treated with different concentrations of [Gly~(14)]-humanin and ?-amyloid protein 1-42.RESULTS: 10 nmol/L [Gly~(14)]-humanin made NSCs resistant to the apoptotic action induced by A?P1-42 and prevented NSCs from death induced by 25 ?mol/L ?-amyloid protein 1-42.The differentiated neural stem cells yield more neuronal cells than that in control groups when 10 nmol/L [Gly~(14)]-humanin was added to the culture media.The number of cells increased and the cultures grown with a manner of floating cell clones likes that cultured in the presence of mitogen when 100 nmol/L [Gly~(14)]-humanin was added to the differentiation culture media.CONCLUSION: The [Gly~(14)]-humanin significantly promoted the proliferation and neuronal differentiation of neural stem/progenitor cells and also inhibited the toxic action of ?-amyloid protein 1-42 on cultured neural stem/progenitor cells.