Inhibition of ICAM-1 expression on endothelial cells in hypoxia/reoxygenation by antisense oligodeoxynucleotides

AIM: To investigate the effect of antisense oligodeoxynucleotides (AS-ODN) on the intercellsular adhesion molecule-1(ICAM-1) expression on endothelial cells in hypoxia/reoxygenation(H/R). METHODS: With cultured glomerular vascular endothelial cell in H/R, the positive percentage of ICAM-1 expression was measured by flow cytometry before and after giving AS-ODN. RESULTS: The ICAM-1 expression did not increase on glomerular vascular endothelial cell in 10 hours hypoxia compared to control group, it increased in 6 hours reoxygenation, and decreased by 40.6% after giving AS-OND. CONCLUSION: AS-ODN may decrease the expression of ICAM-1 on endothelial cells in H/R.     

Expression of phenylalanine ammonia-lyase cDNA from rice in E. coli BL21DE3

AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-β-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78.6 kD on SDS-PAGE analysis, but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique.     

Inducible nitric oxide synthase and arterial pressure regulation

AIM and METHODS:To clarify the role of inducible nitric oxide synthase (iNOS) in the regulation of blood pressure,in the present study, we examined the effect of aminoguanidine (AG), a selective inhibitor of iNOS on the hemodinamical response of Dahl salt-sensitive (DS) and Dahl salt-resistant (DR) rats to low (0.3%) or high (8%) sodium chloride (NaCl) infusion by chronical in vivo hemodynamic experiment, and the effect of NaCl or NaCl plus AG infusion on urinary nitrate (NO₃)/nitrite (NO₂), the end product of nitric oxide (NO), excretion by Greiss Reaction. Furthermore, NOS activity assay was also carried out to probe the effect of NaCl and AG on calcium-dependent or independent NOS activity in renal tissue. RESULTS:1. High or low NaCl-infused DR rats and low NaCl-infused DS rats have no hemodinamical response to AG, however, the hypertensive effect of high NaCl (8%) infusion on DS rats were greatly amplified by co-infusion of AG. 2. Administration of high NaCl significantly elevated the iNOS activity of renal tissue, and greatly increased urinary NO₃/NO₂ excretion. CONCLUSION:Inducible NOS is an important modulator of arterial pressure, especially in case of higher blood pressure.     

Role and regulation of calcineurin in angiotensin Ⅱ-stimulated cardiac myocyte hypertrophy of rats

AIM: To study the role and regulation of calcineurin(CaN) in angiotensin II(AngⅡ)-stimulated cardiacmyocyte hypertrophy of rats. METHODS: Using AngⅡ to induce the cultured cardiac myocyte hypertrophy of rats, and investigating the effect of CaN inhibitor on [³H]-leucine incorporation of AngⅡ-stimulated cardiomyocytes and the regulation of various factors on CaN activity in cardiomyocytes.RESULTS: AngⅡ can stimulate the CaN activity in cultured neonatal rat cardiomyocytes in a dose- and time-dependent manner. In cardiac myocytes incubated with 10, 100, 1000 nmol·L^-1 of AngⅡ for 12h, the CaN activities increased respectively by 13%,57%(P＜0.05) and 228%(P＜0.01) compared with that in non-stimulated cardiomyocytes. The CaN activities in AngⅡ-stimulated cardiomyocytes were significantly inhibited by losartan(50 μmol·L^-1), H₇(50 μmol·L^-1)and Fura-2/AM(4 μmol·L^-1),while no effect was observed with PD98059(50 μmol·L^-1).The [³H]-leucine incorporation in AngⅡ-stimulated cardiomyocytes increased by 46%(P＜0.01) compared with that in control group, which was dramatically inhibited by cyclosporin A(0.5~5μg/mL). CONCLUSIONS: Calcineurin, a Ca²⁺/calmodulin-dependent protein phosphatase, may play an important role in AngⅡ-induced cardiac myocyte hypertrophy. The activation of CaN may dependent on the sustained increases of [Ca²⁺]i and be regulated by some protein kinases (such as PKC,etc.).     

Regulation of‘Tiao Gan Fang Yao’on neuroendocrine-immuno-function of bandage-stressed rats

AIM: To investigate the regulation of ‘Tiao Gan Fang Yao’(TGFY) on neuroendocrine-immuno-function of bandage-stressed rat . METHODS:The stressed rat model was made by bandage. RIA was adopted to measure the function of hypothalamus-pitutary-adrenal gland axis (HPAA) of stressed rat. Meanwhile, the immunity of stressed rat and the regulation of TGFY were observed.RESULTS:Bandage stress increased the contents of serum corticosterone(CORT), and ACTH, and hypothalamus corticotropin releasing hormone (P＜0.01 or 0.05), which suggested that the excitability of HPAA was enhanced. In addition, bandage stress reduced spleen lymphocyte proliferation (P＜0.01) and decreased H₂O₂ releasing from the macrophages significantly (P＜0.01). While TGFY could decrease HPAA excitability of bandage-stressed rat and strengthen its immunity. CONCLUSION:TGFY could regulate disorder of neuroendocrine-immuno-function of bandage-stressed rat.     

Effects of oxidized high-density lipoprotein on membrane fluidity and the expression of lymphocyte function associated antigen-1 of monocyte

AIM: To investigate the effects of oxidized high-density lipoprotein (oxHDL) on membrane fluidity and the expression of lymphocyte function associated antigen(LFA-1) of monocyte. METHODS: The membrane fluidity of THP-1 cells was assayed by fluorescence anisotropy with DPH (1,6-dipheny-1,3,5-hexatriene), a fluorescent probe; The LFA-1 expression on THP-1 cells were assayed by flow cytometry with indirect immunofluorescence.RESULTS: The membrane fluidity of THP-1 cells was reduced by 45% and 52% respectively (P＜0.01) after incubation with 100 μg protein/mL oxHDL and oxLDL for 20 h; oxHDL could stimulated the expression of LFA-1 on THP-1 cells, the positive cell rate and mean fluoresence intensity (MFI) increased by 39% and 45% respectively versus control group (P＜0.01) after incubation with cells for 24 h. CONCLUSIONS: By increasing the expression of LFA-1 on monocyte, oxHDL can promote the monocyte-endothelium adhesion; oxHDL also can reduce the membrane fluidity of monocyte, that will limit the returning of monocytes from subendothelium back to the circulation. This suggested that oxHDL may play an important role in atherogenesis.     

Experimental research on cardiac fibrosis induced by recombinant mouse interleukin-11 in mice

AIM To observe the effect of recombinant mouse interleukin-11 （rmIL-11）injected subcutaneously into mice on heart structure and function and to determine its pro-fibrotic effect. METHODS C57BL/6 mice were randomly divided into experimental group and control group.The mice in experimental group were injected subcutaneously with recombinant mouse IL-11 at the dose of 100 μg·kg^-1·d^-1 for 3 consecutive weeks, while the control group were given equal volume of normal saline in the same way. After the experiment was finished, the parameters of heart function were measured by echocardiography.The heart weight was weighed and the cardiac weight index (CWI) was calculated. HE staining and Masson's trichrome staining were performed to observe the pathological changes and the extent of myocardial fibrosis in mouse myocardia respectively, and the cardiac collagen volume fraction (CVF) was calculated. The expression levels of extracellular matrix proteins in the myocardial tissues of mice, including type Ⅰ collagen, type Ⅲ collagen and fibronectin, were determined by Western blot. RESULTS Left ventricular ejection fraction and left ventricular fraction shortening in experimental group were obviously lower than those in control group (P<0.01), however left ventricular end-diastolic diamension and left ventricular end systolic dimension were significantly higher than those in control group (P<0.05).Compared with control group, the CWI was increased (P<0.01), the myocardial arrangement was disorder, the necrosis of cardiac myocytes was increased, and excessive deposition of collagen was observed in the myocardial tissues in experimental group. Correspondingly, the CVF and protein levels of type Ⅰ collagen, type Ⅲ collagen and fibronectin in the left ventricle in experimental group were increased significantly (P<0.05). CONCLUSION Injection of rmIL-11 into the mice subcutaneously induces fibrogenesis in the heart, which implies that IL-11 is likely a novel pro-fibrotic factor.     

Progress in roles of MNX1 antisense RNA 1 in malignant tumor

MNX1 antisense RNA 1 （MNX1-AS1） is a newly discovered long non-coding RNA （lncRNA）, which is highly dysregulated in various carcinomas and its expression level is closely related to the overall survival and prognosis of patients. MNX1-AS1 regulates the occurrence and development of carcinomas by endogenous competitive adsorption of miRNA, regulating cell cycle, inducing epithelial mesenchymal transformation and activating multiple signaling pathways. The in-depth study of the carcinogenesis of MNX1-AS1 is useful for the early diagnosis, targeted therapy and prognostic assessment of relevant carcinomas. This article reviews the roles of MNX1-AS1 in malignant tumor.     

全子叶生物活性豆腐凝胶特性的研究

通过因子筛选实验,确定了接种量、微生物转谷氨酰胺酶(MTG)用量和水豆比是影响全子叶生物活性豆腐凝胶特性的关键因子。在此基础上,采用响应曲面法(Response SurfaceMethodology,RSM)分别建立了豆腐硬度和持水率的二次多项数学模型,验证了模型的有效性并探讨了上述3个因子的交互作用。从产品成本和凝胶特性综合考虑,选择出最佳配比,即乳酸菌接种量为105cfu/mL,MTG用量为347 U/L,水豆比为8.5,其硬度和持水率实测值分别为44.8g和89.2%,可生产出富含乳酸菌和几乎全部大豆营养的健康豆腐。     

水稻免耕栽培的优化效应与调控技术

水稻免耕栽培主要包括水稻免耕抛秧、免耕直播和免耕插秧等技术。试验示范结果表明:该项技术能提高水稻产量0.9%~5.1%,节省工本,提高纯收入达19.2%~37.0%,并省水节电,增加土壤表层孔隙度,增加土壤速效养分含量。介绍了水稻免耕栽培的调控技术。