Effect of basic fibroblast growth factor on synthesis of C-type natriuretic peptide in cultured human umbilical vein endothelial cells

AIM: To investigate the effect of basic fibroblast growth factor (bFGF) on C-type natriuretic peptide (CNP) production, release and mRNA expression. METHODS: Human endothelial cell cultured;CNP was mea sured by radioimmunoassay method;CNP mRNA expression was determined by RT-PCR technique.RESULTS: bFGF could augment CNP synthesis in human endothelial cells. Compared with control group,25 ng, 50 ng, 100 ng bFGF increased CNP contents in endothelial cells by 88% (P＜0.05), 95% (P＜0.05), 187% (P＜0.01), respectively.100 ng bFGF also stimulated CNP release from cultured human endothelial cell. In addition, 25 ng, 50 ng and 100 ng bFGF stimulated CNP mRNA expression of cultured human endothelial cells in a dose-dependent manner. CONCLUSION: bFGF might regulate CNP synthesis,release and mRNA expression in cultured umbilical human endothelial cells.     

Mechanism of C-type Natriuretic Peptide Regulating Lipid Deposition of Intramuscular Adipocytes in Chickens

The aim of this study was to investigate the regulatory effect and mechanism of C-type natriuretic peptide (CNP) on the proliferation,differentiation and lipolysis of intramuscular preadipocytes (IMPs) in breast muscle of chickens,and lay a foundation on further elucidating the molecular mechanism of intramuscular fat deposition in chickens.The IMPs of breast muscle in Yellow-feathered broilers were used as the model in vitro,the preadipocytes were induced by 10^-7 mol/L CNP.The proliferation,differentiation and lipolysis of IMPs were observed by CCK8,Edu staining,Oil Red O staining and isopropanol extraction,respectively.The concentration of cGMP and glycerin were detected by cGMP and glycerin kit,respectively.The expression of NPRA,NPRB and NPRC genes mRNA was detected by Real-time quantitative PCR.The results showed that compared with control group,the cell proliferation in CNP treatment group was enhanced significantly at 1 and 3 d (P<0.05).The cell differentiation and lipid deposition were extremely significantly decreased at 3 and 6 d (P<0.01),the intracellular cGMP concentration and glycerin concentration in the medium at 3 and 6 d were extremely significantly increased (P<0.01).The expression of NPRB and NPRC genes mRNA in CNP treatment group was extremely significantly or significantly higher than control group (P<0.01;P<0.05),and the expression of NPRA gene mRNA had no significant difference (P>0.05).The results revealed that CNP regulated the lipid deposition of intramuscular adipocytes through NPRB/NPRC-cGMP pathway in chickens.     

Comparison of the amino acid sequences of acorn barnacle lectins showing different inhibitory activities toward the crystal growth of calcium carbonate

ABSTRACT: The amino acid sequence of a D -galactose-binding lectin isolated from the hemolymph of the acorn barnacle Balanus rostratus was determined. The lectin (BRL) ( M _r 120K) is a multimeric protein whose subunit consists of 182 amino acids. The amino acid sequence was compared with those of multiple lectins (BRA-2, BRA-3) from Megabalanus rosa to explore the relationship between the structures and the inhibitory activity toward the crystal growth of calcium carbonate. Although BRL was 46% identical to BRA-2 and 15% identical to BRA-3, the lectin had no inhibitory activity, unlike BRA-2 and BRA-3. Both the number and the localization of acidic amino acid residues and their amide forms were different among them. Observations by scanning electron microscopy revealed modifications in the size and morphology of the calcium carbonate crystals grown in the presence of the lectins.     

绵羊子宫内膜的凝集素结合特性

用５种酶标植物凝集素（ｃｏｎＡ，ＳＢＡ，ＷＧＡ，ＵＥＡ，ＲＣＡ）作探针，研究了间情期和妊娠期绵羊子宫内膜上皮和间质糖复合物的变化。与间情期相比，妊娠期子宫内膜上皮ＣｏｎＡ和ＷＧＡ染色明显增强，特别是ＵＥＡ染色由阴性转变为强阳性，ＳＢＡ染色没有明显变化。在间质石ＢＡ和ＵＥＡ染色由阴性转变为中阳性，而ＣｏｎＡ和ＷＧＡ染色无明显变化。无论间情期或妊娠期，子宫内膜上皮和间质的ＲＣＡ染色均为阴性。子宫内膜上皮和间质糖复合物质和量的变化，可能与子宫各部功能和妊娠有关。     

Effect of combined treatment with GnRHa and GH on linear growth in mid- and late pubertal girls at great bone ages with CPP or EFP and relation to CNP signaling pathway

AIM:To investigate the effect of combined treatment with gonadotropin-releasing hormone analogue (GnRHa) and growth hormone (GH) on the linear growth in mid- and late pubertal girls at great bone ages with central precocious puberty (CPP) or early and fast puberty (EFP), and to determine the relation between C-type natriuretic peptide (CNP) signaling pathway and the accelerative effect of GH on long bone growth in these girls. METHODS:Twenty-two girls were diagnosed as CPP or EFP, whose bone ages were older than 11.5 years with impaired predicted adult height (PAH), and divided into GnRHa treatment group (treated with GnRHa alone, slow-release of triptorelin 60~80 μg/kg every 4 weeks, im) and combined treatment group (treated with GnRHa and GH, 1 U/kg GH every week for 6~7 times, sc). The height, weight and pubertal stage were determined every 3 months. At the beginning and after 6 months of the treatment, the bone age was evaluated and the serum concentrations of amino-terminal pro-C-type natriuretic peptide (NTproCNP), insulin-like growth factor 1 (IGF-1) and procollagen type 1 amino-terminal propeptide (P1NP) were measured. Height velocity (HV), height SD score for bone age (HtSDSBA), PAH and the serum indexes mentioned above were compared at the beginning and the end of the treatment. RESULTS:After 6 months of the treatment, HV, ΔHtSDSBA and ΔPAH of the girls treated with GnRHa+GH were statistically higher than those of the girls given GnRHa alone (P<0.01). Serum concentrations of NTproCNP, P1NP and IGF-1 were not significantly different between the beginning and the end of the 6-month combined treatment. The girls treated with GnRHa alone showed a significant decrease in both serum NTproCNP and P1NP levels (P<0.05) and no significant change of serum IGF-1 level after 6 months of the treatment. CONCLUSION: In the CPP or EFP girls who are in mid- and late puberty and at great bone ages, the combined treatment with GnRHa and GH may accelerate linear growth and improve predicted adult height. This effect of GH is not attributed to the change of serum IGF-1 level, and may be related in part to the acceleration of CNP-mediated long bone growth.     

裙带菜凝集素的分离及性质研究

裙带菜（Undaria pinnatifida)经Tris-HCl缓冲液粗抽提，然后进行硫酸铵分级实验，测定盐析范围为50％硫酸铵饱和度，经50％硫酸铵饱和度分级后，裙带菜凝集素（UPL）纯化倍数为7．7，总活力回收为167％，UPL粗抽提液中糖的质量分数为5．72％。该凝集素在pH为4．0－10．14时均为活性，但在pH为5．0－9．51时活性较高，pH等于4．0时活力保留25％，pH等于10．14时活力保留50％，该凝集素不被D－果糖，葡萄糖，γ－球蛋白所抑制。     

Structure and possible function of <Emphasis Type="Italic">N</Emphasis>-glycans of an invertebrate C-type lectin from the acorn barnacle <Emphasis Type="Italic">megabalanus rosa</Emphasis>

ABSTRACT:   A C-type lectin (BRA-2) isolated from the acorn barnacle Megabalanus rosa , which was a glycoprotein having an N -linked sugar chain, was deglycosylated by N -glycopeptidase F. The structure of the released sugar chains was determined by a 2-D mapping method after derivatization with a fluorescent reagent, 2-aminopyridine, to be Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc and Manα1-6(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc. The structures were confirmed by matrix-assisted laser desorption ionization mass spectrometry and a comparison with authentic sugar chains by high-pressure liquid chromatography. Various properties of BRA-2 were examined before and after deglycosylation. The susceptibility of BRA-2 to protease digestion was increased by deglycosylation. However, the inhibitory activity toward calcium carbonate crystallization as well as the hemagglutinating activity of deglycosylated BRA-2 was significantly decreased. These results suggest that the sugar chains of BRA-2 are important to both its structural stability and its function.     

重组乳酸乳球菌对模拟胃肠道的耐受力研究

[目的]研究重组菌株在不同p H胃液和胆盐浓度肠液中的生存情况,验证重组菌株能否在肠道中发挥作用,为其制备基因工程活载体疫苗、新型免疫型微生态制剂及功能发酵食品提供理论依据。[方法]人工模拟人体胃肠道环境,研究表达金黄色葡萄球菌纤黏蛋白原凝集素A(Clf A)的重组乳酸乳球菌MG1363/p MG36c-clf A菌株在不同p H胃液和胆盐浓度的肠液中的耐受力。[结果]重组菌株在p H 3.0的人工胃液中处理2 h后,存活率约为88.6%;在0.3%胆盐浓度的人工肠液中处理2和4 h后,存活率约为83.6%和68.2%。[结论]重组菌株对胃肠道环境有较强的耐受力,适合制备基因工程活载体疫苗、新型免疫型微生态制剂及功能发酵食品。     

植物凝集素SMLII 基因启动子的克隆及序列分析

根据已克隆的丹参植物凝集素SMLII基因cDNA序列,利用染色体步移技术克隆出SMLII 启动子区序列,并进行生物信息学预测分析该基因的功能,结果得到了该基因上游1023 bp的启动子序列（GenBank AccessionNo: KF193867）。分析表明,在该序列中不但含有真核生物典型的核心启动子区TATA-box、CAAT-box,而且含有一些特有的上游启动子元件ABRE、ARE、ASF1MOTIFCAMV、HSE、IBOXCORE、WBOXATNPR1等重要的顺式调控元件及一些光反应相关元件。     

Characterization of glycoconjugates and sialic acid modification in the olfactory bulb of the Chinese fire-bellied newt (Cynops orientalis)

Diverse glycoconjugates are expressed in the vertebrate olfactory bulb and serve as guidance cues for axons of nasal receptor neurons. Although the involvement of glycoconjugates in the segregation of the olfactory pathway has been suggested, it is poorly understood in salamanders. In this study, lectin histochemistry was used to determine glycoconjugate distribution in the olfactory bulb of the Chinese fire-bellied newt (Cynops orientalis). Succinylated wheat germ agglutinin (sWGA), Ricinus communis agglutinin-I and Lens culinaris agglutinin showed different bindings in the nerve fibre layer or glomerular layer, or both, between the main and accessory olfactory bulbs. We then investigated the lectin-binding pattern after the removal of terminal sialic acids using neuraminidase. Desialylation resulted in a change in the binding reactivities with seven lectins. Wheat germ agglutinin, sWGA, soybean agglutinin (SBA) and peanut agglutinin showed different degrees of binding between the main and accessory olfactory bulbs. In addition, SBA showed a heterogeneous labelling of glomeruli in the rostral region of the main olfactory bulb. Our results suggest that terminal sialic acids mask the heterogeneity of glycoconjugates in the olfactory bulb of C. orientalis.