Macaca irus bone mesenchymal stem cells differentiate into neuron-like cells induced by cryptotanshinone in vitro

AIM: To determine the optimal protocol and condition in which macaca irus mesenchymal stem cells (MSCs) are induced to differentiate into neuron-like cells by cryptotanshinone in vitro. METHODS: MSCs from macaca irus bone marrow were generated in vitro and induced with cryptotanshinone. The morphological changes of MSCs were evaluated by microscope. The positive percentages of neurofilament (NF), neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression were measured by immunocytochemistry with ABC staining. RESULTS: The result showed that MSCs were positive for CD29, CD44, CD105, CD166, and negative for CD34, CD71, CD80 and CD86. After induced with cryptotanshinone, MSCs began to display neuronal morphologies, such as contracted multipolar cell body and formed extensive networks. The percentages of positive NSE, NF expression were 68.3%±3.5%, 70.3%±1.5%， respectively. CONCLUSION: Macaca irus MSCs could be induced to differentiate into neuron-like cells in vitro by cryptotanshinone and might be applied in cell transplantation and gene therapy in nervous system disorders.     

Significance of expression of extracellular matrix metalloproteinase inducer and endothelin converting enzymes in lymphoid metastasis and prognosis in non-small-cell lung carcinoma

AIM: To investigate the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) and endothelin converting enzymes (ECE) in non-small-cell lung carcinoma (NSCLC), and their relationship with lymphoid metastasis and prognosis.METHODS: Expression of EMMPRIN and ECE in 77 cases of patients with NSCLC was detected immunohistochemically.The relationship of the expression of EMMPRIN and ECE with tumor size, histological type, differentiation, lymphoid metastasis, clinical stage, and prognosis were analyzed.RESULTS: The expressive rate of EMMPRIN and ECE was 66% and 45%, respectively.The expression of EMMPRIN and ECE was associated positively with lymphoid metastasis (both P<0.01, r=0.371 and 0.467, respectively), and inversely with survival time (P<0.01 and P<0.01).No relationship was found in the expression of EMMPRIN and ECE with tumor size, histological type, and differentiation (P>0.05).The expression of EMMPRIN was associated with the expression of HGF in NSCLC (P<0.05, r=0.243).CONCLUSION: The expression of EMMPRIN and ECE is associated with lymphoid metastasis and prognosis in NSCLC.Overexpression of EMMPRIN and ECE implies infavourable prognosis in NSCLC.     

Effect of diet with high fats and cholesterol on PPARα gene expression and body cholesterol level in mice

AIM:To investigate the effects of dietary fats and cholesterol on liver PPARα gene expression and body cholesterol (Chol) level in C57BL/6J mice. METHODS:The animals (n=75) were randomly divided into five groups and respectively received formula mash for 6 weeks. RESULTS:As compared to Chol diet, Chol+PUFA diet produced significantly higher liver cholesterol (P<0.01), serum total cholesterol (TC), focusing on HDL-C. While Chol+MUFA diet resulted in unchanged serum TC and lower liver cholesterol (P<0.01). Chol+SFA diet rsulted in higher liver cholesterol (P<0.01) and serum TC, focusing on LDL-C. Furthermore, Chol+PUFA diet increased the mRNA and protein content of PPARα (P<0.01) in liver, while Chol+MUFA and Chol+SFA diets decreased the mRNA content of PPARα significantly (P<0.01). CONCLUSIONS:These results indicated that addition of fats containing PUFA to diet high in cholesterol increased PPARα mRNA and protein expression, addition of fats containing MUFA or SFA to diet high in cholesterol decreased PPARα mRNA expression. The change of PPARα gene expression may further affect body cholesterol level.     

Role of neutrophil elastase and myeloperoxidase in asthma of rats

AIM:To observe the changes of polymorphonuclear leukocytes (PMN), neutrophil elastase (NE) and myeloperoxidase (MPO) expression in rat asthma. METHODS:Eighteen rats were randomly divided into two groups on average, including asthma group and control group. The rat model of asthma was established by the ovalbumin (OVA) challenge methods. Blood PMN were isolated andpurified. The protein expression of MPO were detected by immunohistochemistry and chromatometry techniques. NE was detected by ELISA. RESULTS:(1) Immunohistochemistry showed that the levels of MPO expression around bronchus and in purified PMN in asthma group were significantly increased compared with those in control group (P<0.01). Activities of MPO were significantly increased in bronchoalveolar lavage fluid (BALF) and lung tissue in asthma group compared with those in control group (P<0.05, P<0.01). (2) Levels of NE were significantly increased in BALF and PMN in asthma group compared with control group (P<0.01). (3) Number of PMN were significantly higher in BALF, bronchus and lung tissue in asthma group than those in control group (P<0.01). CONCLUSION:Levels of PMN counting and expression of MPO and NEare significantly increased in experimental asthma rats. PMN may be involved in inflammation in asthma via increasing the levels of NE and MPO.     

Effect of mitochondrial ATP sensitive potassium channels opening on translocation of protein kinase C epsilon in cultured adult rat ventricular myocytes

AIM: To investigate the effects of mitochondrial ATP sensitive potassium (MitoKATP) channel opening on translocation of protein kinase C epsilon (PKCε) and the relationship between the translocation of PKCε and the production of reactive oxygen species. METHODS: The expression of PKCε in cultured adult rat ventricular myocytes was investigated with immunofluorescence and Western blotting techniques. RESULTS: （1） Diazoxide, a selective MitoKATP channel opener, caused a significant translocation to myofibrillar-like structures in cultured adult rat ventricular myocytes. (2) The N-2-mercaptopropionylglycine (MPG), a free radical scavenger, partly inhibited the translocation of PKCε caused by diazoxide. (3) Chelerythrine, a selective protein kinase C (PKC) inhibitor, completely blocked the translocation of PKCε caused by diazoxide. CONCLUSION: Opening of MitoKATP channel might activate PKCε and make it translocation to myofibrillar-like structures. PKCε activation occurs downstream of MitoKATP channel, and might be caused by production of reactive oxygen species after opening of MitoKATP channel.