Folic acid treatment regulates C2C12 myoblast diferentiation via JNK/p38 MAPK signaling pathway / 中国病理生理杂志

AIM:To observe the effect of folic acid(FA)on C2C12 myoblast proliferation and differentia-tion,and to explore its mechanism.METHODS:During the proliferation stage,C2C12 myoblasts were treated with vari-ous concentrations of FA(0,2.5,5,10 and 20 μmol/L).The cell status was observed under a microscope,cell viability was detected using the MTT method,and cell proliferation was assessed using the EdU method.In the differentiation stage,C2C12 cells were divided into control(Ctrl)group(0 μmol/L FA)and FA group(10 μmol/L FA).On day 2 or 4 of differentiation,immunofluorescence staining and Western blot were employed to detect the expression of myoblast differen-tiation-related proteins,myoblast determination protein 1(MyoD),myogenin(MyoG)and myosin heavy chain(MyHC).The myotubule formation in each group was analyzed.On day 4 of differentiation,C2C12 cells were treated with FA for 0,1,3 and 6 h,and the protein levels of p-JNK,JNK,p-p38 MAPK and p38 MAPK at each time point were detected by Western blot.Additionally,C2C12 cells after 4-day differentiation were divided into Ctrl group,FA group,FA+ SP600125(specific inhibitor of JNK)group,and FA+SB203580(specific inhibitor of p38)group.The cells in FA+ SP600125 and FA+SB203580 groups were treated with 10 μmol/L SP600125 or SB203580 for 1 h,followed by treatment with 10 μmol/L FA for 24 h.The cells in FA group were treated with 10 μmol/L FA for 24 h,while the cells in Ctrl group were left untreated.The protein levels of p-JNK,JNK,p-p38 MAPK,p38 MAPK and MyHC were detected by Western blot.RESULTS:(1)Compared with 0 μmol/L FA group,the number of the cells in other concentration groups in-creased,cell viability was raised(P＜0.05 or P＜0.01),and the rate of EdU positive cells increased(P＜0.05).(2)Com-pared with Ctrl group,the expression levels of MyoD,MyoG and MyHC in FA group were increased(P＜0.05),and the myotube fusion index was raised(P＜0.05 or P＜0.01).(3)Compared with 0 h group,the ratios of p-JNK/JNK and p-p38 MAPK/p38 MAPK were elevated after FA treatment for 1,3 and 6 h(P＜0.05 or P＜0.01),and showed a trend of gradual increase with the extension of treatment time.(4)After FA treatment,the ratios of p-JNK/JNK and p-p38 MAPK/p38 MAPK,and the expression of MyHC were elevated(P＜0.01).Treatment with SP600125 decreased the ratio of p-JNK/JNK and the expression of MyHC(P＜0.05),while SB203580 intervention cut down the ratio of p-p38 MAPK/p38 MAPK and the expression of MyHC(P＜0.05 or P＜0.01).CONCLUSION:Folic acid can promote the differentiation of C2C12 myoblasts by activating the JNK/p38 MAPK signaling pathway.

Effect of Liraglutide on Toll-like Receptor 4 and Na+/K+-ATPase Activity in Proximal Tubular Epithelial Cells of Rats Induced by High Glucose / 医药导报

Objective To explore the protective effect of liraglutide ( Li) on the proximal tubular cells ( PTECs) of rats induced by model control. Methods The PTECs of rats were obtained by primary culture and divided into normal control group ( NC) , model control group ( HG) , HG+low-dose Li group, HG+medium-dose Li group, and HG+high-dose Li group ( n-6 in each group) . After 72 h of incubation, the activity of Na+/K+-ATPase in PTECs was measured by the liquid scintillation counter. Protein expression of Toll-like receptor 4 ( TLR4 ) was measured by Western blotting. Interleukin-6 ( IL-6 ) , tumor necrosis factor-α (TNF-α) and plasminogen activator inhibitor-1 (PAI-1) in culture supernatants were measured by the enzyme linked immunosorbent assay (ELISA). Results Compared with NC, activity of Na+/K+-ATPase [(2 737. 88±317. 22)μmol·g-1 ·h-1 ] and the protein expression of TLR4 in PTECs, IL-6, TNF-α and PAI-1 in culture supernatants were significantly increased in HG (P<0. 05, or P<0. 01). Compared with HG, activity of Na+/ K+-ATPase and the protein expression of TLR4 in cells, IL-6, TNF-αand PAI-1 in culture supernatants reduced significantly in HG+medium-or high dose Li groups (P<0. 05, or P<0. 01). Conclusion Liraglutide can inhibit the expression of inflammatory cytokines and stabilize the Na+/ K+-ATPase’ s activity in PTECs in high glucose environment. Therefore, it may play a role in kidney protection.

Analysis on clinical characteristics of 10 patients with granulocytic sarcoma / 白血病·淋巴瘤

Objective To explore the clinical characteristics,diagnosis,therapy and prognostic features of granulocytic sarcoma (GS).Methods Retrospective analysis was used to analyse 10 cases who were diagnosed with GS.Results Five patients were diagnosed with primary GS.GS was accompanied by APL in one case,CML in 3 cases and MDS in one case.Patients were treated with resection and chemotherapy,or imatinib treatment.Follow-up data were available from 9 patients,with 2 patients being still alive.The survival of one patient who received high-dose cytarabine chemotherapy was 66 months.The other 7patients died of tumor-related diseases,and their survival ranged from 4 to 17 months.Conclusion GS is a rarely diagnosed disease.A correct diagnosis of GS depends on detailed morphological examination and immunohistochemical study.The clinical outcome of patients is poor,and AML-type chemotherapy is the proper treatment for GS.High-intensity chemotherapy and hematopoietic stem cell transplantation might improve long-term survival.

Bone marrow mononuclear cell transplant therapy in mice with CCl4-induced acute liver failure.

BACKGROUND/AIMS: Stem cell transplantation has theoretical potential for the treatment of certain liver diseases. However, the use of bone marrow mononuclear cells as a therapy for liver disease has received little attention. The present study was to examine whether bone marrow mononuclear cells might be useful in the management of acute liver failure in an animal model. MATERIALS AND METHOTS: Bone marrow mononuclear cells were harvested from BALB/c mice and then labeled with the fluorescent dye PKH26. The labeled cells were subsequently infused into the tail veins of mice in which hepatic injury had been induced by CCl4 toxicity. After transplantation, the labeled cells in the liver were studied by fluorescent microscopy, and the levels of proliferating cell nuclear antigen and albumin were quantified in bone marrow mononuclear cell-treated and untreated groups. Serum aminotransferase activity was also monitored at various time points post-liver injury. RESULTS: Transplanted bone marrow mononuclear cells labeled with PKH26 were found to populate the damaged liver around the portal and centrolobular regions, and they appeared to differentiate into albumin-producing hepatocyte-like cells. Animals that received bone marrow mononuclear cells also showed a trend toward improved liver enzymes as well enhanced survival rates, relative to controls. CONCLUSIONS: These findings suggest that systemically delivered bone marrow mononuclear cells may relocate to and be retained by the injured liver; transplantation of bone marrow mononuclear cells showed an overall beneficial effect in a murine model of acute liver failure.

Expression and clinical implication of HIF-1α mRNA in mononuclear cells of bone marrow of multiple myeloma / 白血病·淋巴瘤

Objective To analyze the correlation between the expression of HIF-1α in multiple myeloma (MM) and clinical indexes in order to illustrate the expression and implication of HIF-1α gene in MM. Methods RQ-PCR method was used to amplify HIF-1α mRNA of bone marrow mononuclear cells isolated from 28 cases of MM patients and the control group. β-actin was used as internal standard. HIF-1α mRNA expression was analyzed by SDS software and the ratios of HIF-1α/ β-actin were calculated. Results The relative expression level of HIF-1α mRNA in MM bone marrow mononuclear cells was 12.68 times as that in control group. HIF-1α mRNA was positively correlated with β2-MG (r =0.575, P =0.000), ESR (r =0.522,P =0.000), LDH (r=0.286, P=0.044) and CRP (r =0.356, P =0.011). There was a negative correlation between HIF-1α mRNA and Hb (r =-0.556, P =0.000). Conclusion The expression of HIF-1α mRNA was up-regulated and HIF-1α was related to a number of clinical indexes. HIF-1α may be used to estimate the progress of MM and hopeful to be a new molecular target in cancer therapy.

Expressions of BAG-1 and NF-?B P65 in the mononuclearcell of multiple myeloma patients / 中国临床药理学与治疗学

AIM:To investigate the expressions of BAG-1 and NF-?B P65 in the multiple myeloma(MM)and approach the effects of BAG-1 and NF-?B P65 in the pathogenesis of MM and their clinical significance,which would be useful to find a theoretical basis for the treatment of MM.METHODS:The patients diagnosed MM in Yijishan hospital of wannan medical college from 2006.10 to 2008.10 as the experimental group,while the bone trauma patients selected as the control group.The mononuclearcells of bone marrow in the patients were extracted and the BAG-1and NF-?B P65 mRNA RT-PCR were detected.RESULTS:The expression of BAG-1 mRNA(71.4%)in the MM patients was higher than that in the control group(P=0.007).The expression of NF-?B P65 mRNA(75.0%)in the MM patients was higher than that in the control group(P=0.004).There was a positive correlation between the BAG-1 and NF-?B P65 mRNA.The tumor load was high in the MM patients with overexpression of BAG-1 and NF-?B P65,which was ralated to a number of clinical parameters indexes.BAG-1 and NF-?B P65 were positive expressed of MM tumor load,relatived with a number of clinical indicators.CONCLUSION:The expressions of BAG-1 and NF-?B P65 in MM are high,there are no difference between the experimental group and control group.BAG-1 and NF-?B P65 were related to a number of clinical indexes.BAG-1 and NF-?B P65 may be involved in the occurrence and the development of disease,and they can be used as one of the prognosis indexes.