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Objective To investigate the effect of cyclic stretch on Src and Runt-related transcription factor 2 (RUNX2), and their pivotal roles in migration of vascular smooth muscle cells (VSMCs). Methods The 5% cyclic stretch (to simulate normotensive physiological condition) or 15% cyclic stretch (to simulate hypertensive pathological condition) was applied to VSMCs by FX-5000T system. Western blotting was used to detect the expression of RUNX2 and phosphorylation of Src in VSMCs. The Ingenuity Pathway Analysis (IPA) bioinformatic software was used to analyze the potential regulatory effect of Src on RUNX2. Small interfering RNA (siRNA) was transfected to decrease the expression of RUNX2. Src inhibitor-1 was used to repress Src kinase activity; Wound-healing assay was applied to detect VSMC migration. Results Compared with 5% cyclic stretch, 15% cyclic stretch significantly increased RUNX2 expression in VSMCs. Under both static and 15% cyclic stretch conditions, VSMC migration was significantly inhibited after reducing RUNX2 expression with siRNA transfection. IPA indicated that Src kinase might be the upstream modulator of RUNX2, and Western blotting validated that RUNX2 expression was significantly decreased after inhibiting Src. Furthermore, under 15% cyclic stretch, Src inhibitor-1 markedly repressed RUNX2 expression and VSMC migration.Conclusions High cyclic stretch increased phosphorylation of Src kinase and expression of RUNX2, which subsequently induced VSMC abnormal migration. Exploring the mechanobiological mechanism of VSMC migration regulated by cyclic stretch may contribute to further revealing the mechanism of vascular physiological homeostasis and vascular pathological remodeling, as well as providing new perspective for the translational research of vascular remodeling upon hypertension.
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Objective@#To investigate the expression of TTK (tyrosine and threonine protein kinase) in the process of colorectal cancer (CRC) development and its relationship to prognosis in CRC patients.@*Methods@#Colitis-associated colon cancer model was induced by azoxymethane (AOM) and dextran sulfate sodium (DSS) in C57BL/6 mice. Mice at four different stages of colon cancer development were obtained, named AD1 (inflammation of the recovery), AD2 (mild dysplasia), AD3 (adenoma) and AD4 (adenocarcinoma), as well as negative control (no treatment). The expression of TTK was measured by real time fluorescent quantitative PCR (qPCR) and immunohistochemical staining in mouse colon tissues and 24 pairs of CRC specimens. The relationship between TTK and prognosis was analyzed in a set of CRC genome-wide gene expression microarray data that was obtained from Gene Expression Omnibus (GEO) of National Center for Biotechnology Information (NCBI).@*Results@#The genome-wide microarray data from mouse AOM-DSS model indicated that the expression of TTK mRNA was gradually elevated during the development of colon cancer. The subsequent qPCR results showed that TTK mRNA levels in negative control, AD1, AD2, AD3 and AD4 groups were 1.05±0.42, 1.10±0.03, 1.38±0.15, 1.33±0.17 and 2.12±0.22, respectively. And TTK expression in AD2, AD3 and AD4 groups were significantly higher than that in negative control (P<0.05). The protein expression of TTK showed by immunohistochemical staining had similar tendency as the results of TTK mRNA. Besides that, the TTK mRNA levels in tumor tissues (0.71±0.10) from 24 CRC patients were significantly higher than those in paired adjacent normal tissues (0.18±0.04; P<0.001). The positive expression rate of TTK protein in 5 pairs of CRC clinical samples was 80.0%, and it was significantly higher than that in adjacent normal tissues (30.8%, P=0.014). Furthermore, according to a public transcriptomic data (GSE17536), the high levels of TTK were associated with poor prognosis in CRC patients.@*Conclusions@#Elevated expression of TTK is related to colonic carcinogenesis in both of mouse model and human CRC samples. TTK is a poor prognostic factor in CRC.
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Objective To explore the effects of ZNF330 on erythroid differentiation of K562 cells and underlying mechanism .Methods Realtime PCR was performed to detect the expression of ZNF 330 in K562 cells induced by hemin .After CD34 +cells being infected by the recombination lentivirus ZNF 330-RNAi, Realtime PCR was applied to detect the expression of CD235a and γ-globin.The luciferase report assay was performed to examine if ZNF 330 could act as a trans-acting factor in 293T/17 cells.Co-Immunoprecipitation (Co-IP) was applied in 293T/17 cells to detect the interaction between ZNF 330 and ZNF408 which was involved in mRNA degradation .Results The ex-pression of ZNF330 was up-regulated after hemin treatment .The expression of CD235a andγ-globin decreased after inhibition expression of ZNF 330 had no effect on report gene .Co-Ip in two ways confirmed the direct binding be-tween ZNF330 and ZNF408 .Conclusions ZNF330 can promote erythroid differentiation , and a possible mecha-nism is that ZNF330 inhibits the function of ZNF 408 , a factor that is involved in mRNA degradation , through the interaction between the two proteins .
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Objective To study the stability of sodium nitroprusside for injection in 5% glucose injection and 0 .9%sodium chloride injection ,and provide reference for clinical rational drug use .Methods The content of sodium nitroprusside for injection in different dissolvants(5% glueose ,sodium chloride injection) within 26 h was determined by HPLC .The change of micro particle was measured by light blockage method in accordance with China pharmacopeia(2010 edition) ,and the appearance and UV visible absorption spectroscopy and pH ,the osmotic pressure of the mixtures were observed as well .Results The changes in appearnace ,UV visible absorption spectroscopy ,pH value ,content and microparticle were not significant .Conclusion Sodium nitroprusside for injection were stable within 26 h after mixing with different dissolvants (5% glueose ,sodium chloride injection) .
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Objective:To investigate the best compatibility plan for nitroprusside sodium for injection. Methods: An L16 (43 × 26 ) orthogonal design was used with four factors:temperature, solvent, solvent volume and time. The content of sodium nitroprusside was detected by HPLC, the insoluble particles were observed by a GWF-5J type particle analyzer, pH was detected by a PHS-2C preci-sion acidity meter, and a comprehensive scoring method was used to optimize the compatibility plan. Results:The optimized compati-bility plan was as follows:50mg sodium nitroprusside was dissolved in 250ml sodium chloride injections, avoiding light and used up in 26h. Conclusion:The plan provides scientific basis for the clinical medication.
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Objective:To evaluate the compatible stability of sodium nitroprusside for injection ( SNP) in two kinds of solutions (5% glucose and sodium chloride injections) . Methods:A, B, C and D solution of SNP was respectively prepared according to the concentration of clinical use. The stability of SNP solution was studied in different environment with different storing time, and the as-sessment indices included the appearance, insoluble particles, pH value, ultraviolet-visible absorption spectrum and SNP content. Re-sults:The SNP solutions in 5% glucose and sodium chloride injections were stable in 26 hours in photophobic condition at 20℃ and 35℃. Under an incandescent lamp with or without lightproof, the solutions were very unstable. Conclusion:Both sodium chloride and 5% glucose injections can be used as the solvent for SNP with 26h stability. SNP injections prepared and used in clinics should be pre-served away from light to ensure the safety and effectiveness.
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Twenty kinds of amino acids are simplified into 3 types: hydrophobic amino acids (H), hydrophilic amino acids (P) and neutral amino acids (N). Each residue is reduced to a bead which locates in the position of the C?琢 atom. The off-lattice model is adopted and the relative entropy is used as a minimization function to predict the tertiary structure of a protein. A new contact intensity function is given to consist with protein design research based on the relative entropy. Testing on several real proteins from Protein Data Bank (PDB) shows the good results obtained with the model and method. The root mean square deviations (RMSD) of the predicted structures relative to the native structures range from 0.30 to 0.70 nm. A foundation for studying protein design using the HNP model and the relative entropy was made.