Obestatin对血浆ghrelin和nesfatin-1水平及胃排空的影响

目的:探讨侧脑室注射obestatin对大鼠血浆酰基化ghrelin、去酰基化ghrelin、nesfatin-1水平的影响以及对胃排空的调控。方法:侧脑室注射obestatin,采用酶免疫测定(EIA)法检测血浆酰基化ghrelin、去酰基化ghrelin、nesfatin-1水平以及胃排空率的变化。结果:侧脑室分别注射0.1、0.3或1.0 nmol obestatin,大鼠血浆酰基化ghrelin、去酰基化ghrelin以及nesfatin-1水平无显著改变(P0.05),且酰基化ghrelin与去酰基化ghrelin比率无显著改变(P0.05);侧脑室注射obestatin,大鼠摄食量无显著改变,但胃排空率明显增加(P0.05);胃排空率明显延迟(P0.05)。与侧脑室注射1.0 nmol Obestatin组相比,注射1.0 nmol Obestatin+CRF,大鼠摄食量无显著改变,胃排空率明显延迟(P0.05)。各组摄食量及进入十二指肠内食物量无明显差异(P0.05)。结论:中枢obestatin促进大鼠的胃排空,可能与h/r CRF通路有关。     

姜辣素对链霉素诱导糖尿病大鼠ghrelin表达及胃轻瘫的影响

目的:探究姜辣素对链霉素诱导糖尿病大鼠胃肠功能的影响。方法:链霉素诱导糖尿病大鼠连续灌胃姜辣素(100,200,400 mg/kg)28天,检测胃部超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性、谷胱甘肽(GSH)和丙二醛(MDA)含量以及胃排空。结果:糖尿病大鼠SOD、CAT以及GSH水平降低(SOD:6.29±1.03 vs.3.41±1.21;CAT:27.43±5.27 vs.10.52±2.37,GSH:1091.27±170.09 vs.685.07±75.24,P0.05),MDA水平增高(9.79±2.41 vs.46.38±12.59,P0.05),姜辣素灌胃后SOD、CAT以及GSH水平增高(SOD:3.41±1.21 vs.4.36±1.01,5.62±1.18,7.05±1.48,6.48±1.82;CAT:10.52±2.37 vs.15.27±4.59,19.29±5.42,23.79±6.35,GSH:10.52±2.37 vs.15.27±4.59,19.29±5.42,23.79±6.35,P0.05-0.01),MDA水平降低(46.38±12.59 vs.34.61±9.27,28.01±8.34,19.17±5.19,P0.05-0.01);糖尿病大鼠胃ghrelin水平下降(381.26±94.37 vs.195.07±57.42,P0.01)血浆ghrelin水平上升(76.86±21.81 vs.108.83±27.75,P0.05)胃ghrelin m RNA表达增多,给予中高剂量姜辣素后胃ghrelin水平上升(195.07±57.42 vs.301.43±81.24,328.93±76.59,P0.05~0.01)胃ghrelin m RNA表达减少,呈量效依赖关系;糖尿病大鼠胃排空降低(82.24±19.74 vs.45.37±11.23,P0.01),给予姜辣素后胃排空增强(45.37±11.23 vs.52.43±15.42,49.89±9.84,74.39±20.79,P0.05-0.01)。结论:姜辣素通过抗氧化性改善糖尿病胃轻瘫。     

中枢nesfatin-1对大鼠夜间摄食和胃排空的影响

目的:观察中枢nesfatin-1对大鼠夜间摄食和胃排空的影响。方法:大鼠经腹腔注射硫酸仲丁巴比妥(100~150 mg/kg)麻醉,侧脑室、第四脑室或小脑延髓池注射nesfatin-1或CRF受体拮抗剂astressin-B或astressin2-B,观察对摄食、胃排空的影响。结果:侧脑室注射nesfatin-1后大鼠第3-6 h夜间进食量(t=3.05~3.58,P0.01)和3 h和6 h的累积进食量(t=5.90~12.1,P0.01)明显减少,nesfatin-1的该抑制效应可被预先侧脑室注射astressin-B或astressin2-B阻断(t=1.06~2.22,P0.05)。第四脑室或小脑延髓池注射nesfatin-1后大鼠夜间摄食量在第1h就明显减少(t=2.59~6.26,P0.05~0.01),持续减少至5-6h(t=1.69~7.42,P0.05~0.01)。侧脑室注射不同剂量nesfatin-1(0.05或0.5μg)20 min后GE率明显降低,且随注射剂量增大,GE率越低(t=3.25~4.67,P0.01)。若预先给予大鼠CRF受体拮抗剂astressin2-B(30μg)再注射nesfatin-1(0.5μg),nesfatin-1抑制大鼠胃排空效应明显减弱(t=2.45~2.85,P0.05)。禁食24 h后再喂食2 h,大鼠下丘脑中nesfatin-1表达明显增加(t=2.87,P0.05),禁食24 h后血浆nesfatin-1水平明显降低(t=1.51,P0.05)。结论:Nesfatin-1抑制摄食作用可能由nesfatin-1和CRF2信号系统共同调节。     

胃转流术后远端肠黏膜形态及生长因子表达的变化

目的:探讨胃转流术后远端肠管黏膜的适应性变化及生长因子的表达情况。方法:将8周龄的Wistar大鼠随机分为对照组、假手术组和胃转流术组,术后8周取吻合口远端肠管行常规病理切片检查,测量肠黏膜厚度和绒毛高度,采用免疫荧光法检测肠粘膜中表皮生长因子(EGF)和胰岛素样生长因子-1(IGF-1)的表达。结果:胃转流术组黏膜厚度(672±39与500±31μm,P<0.01)和绒毛高度(445±19与342±15μm,P<0.01)均显著高于假手术组(P<0.01),而对照组和假手术组间均无显著差异。与假手术组比较,胃转流术组肠粘膜中EGF和IGF-1的表达显著升高(P<0.01),而对照组和假手术组间无显著差异。结论:胃转流术后远端肠管粘膜发生适应性增生,同时伴随着EGF和IGF-1表达水平的升高。     

顺铂化疗对下丘脑、血浆ghrelin和orexin 表达的影响

目的:观察顺铂化疗对下丘脑、血浆ghrelin、orexin表达和摄食量的影响。方法:Real-time PCR、ELISA法观察顺铂对大鼠下丘脑、血浆ghrelin、orexin表达及摄食量的影响;19名接受顺铂经导管动脉灌注化疗(TAI)的肝细胞患者(HCC),ELISA法检测化疗前和化疗后血浆ghrelin、orexin的变化,用直观类比标度(VAS)(0-10)评估食欲和摄食量。结果:每日腹腔注射顺铂6 mg/kg,1-5 d大鼠摄食量均显著减少(P0.05),且1-4 d血浆酰化ghrelin显著降低(P0.05),5d时浓度仍低于对照组,但无统计学意义。血浆非酰化ghrelin和总的血浆ghrelin没有明显变化(P0.05),而1-5天血浆orexin水平均明显降低(P0.05);顺铂注射1 d后,大鼠下丘脑ghreilin和orexin的mRNA表达量均显著减少(P0.05),ghrelin mRNA变化持续3 d,orexin mRNA在化疗后5 d仍低于对照组(P0.05);肝细胞癌患者化疗后1至8 d的摄食量明显降低,1 d和2 d时的血浆酰化ghrelin显著低于化疗前水平(P0.05)。3 d时逐渐恢复,化疗后3 d、4 d和7 d时血浆酰化ghrelin浓度与化疗前无统计学差异(P0.05)。血浆非酰化ghrelin和总的血浆ghrelin没有明显变化(P0.05);化疗后1~4 d时血浆orexin浓度均显著降低(P0.05),化疗后7 d时orexin基本恢复到化疗前水平(P0.05)。结论:顺铂可降低大鼠下丘脑和血浆ghrelin、orexin的mRNA表达,HCC的TAI会降低血浆酰化ghrelin、orexin、和摄食量。     

下丘脑Nesfatin-1/NUCB2对糖尿病小鼠摄食的影响

目的:探讨下丘脑神经肽NUCB2与Tsumura Suzuki(TS)多基因突变2型糖尿病(T2DM)小鼠摄食过多的关系。方法:将动物分为Tsumura Suzuki糖尿病(TSD)小鼠、正常小鼠;监视器监测小鼠摄食量;分析血生化指标;定量RT-PCR分析摄食相关神经肽m RNA表达水平;放射免疫分析法检测nesfatin-1蛋白水平。结果:与年龄匹配的TSN小鼠相比,TSD小鼠在1月龄就存在体重增加(P<0.05)和高瘦素血症(P<0.05),3-12月龄出现贪食(P<0.05)、高血糖(P<0.05)、高血脂(P<0.05)和高胰岛素血症(P<0.05),且3-12月龄时厌食肽nesfatin-1前体核连蛋白2(NUCB2)m RNA和nesfatin-1蛋白水平均显著降低(P<0.05~0.01);TSD小鼠下丘脑甘丙肽、黑色素浓集素、神经肽Y及前黑素细胞皮质素原m RNA水平也有显著改变(P<0.05)。结论:下丘脑NUCB2介导信号通路破坏可能导致TSD小鼠摄食过多。     

瓦氏雅罗鱼生殖洄游过程中离子调节相关生理变化研究

为了解达里湖瓦氏雅罗鱼(Leuciscus waleckii)生殖洄游过程中血清离子调节相关生理变化, 对比了达里湖和贡格尔河瓦氏雅罗鱼血清离子(Na+、K+、Cl?、Ca2+和Mg2+)水平, 鳃、肠和肾组织Na+/K+-ATPase和鳃Ca2+/Mg2+-ATPase活性、血清催乳素(PRL)、生长激素(GH)和类胰岛素生长因子-1(IGF-1)水平及鳃组织结构差异; 并利用实验生态学方法, 研究达里湖中瓦氏雅罗鱼转入贡格尔河水24h后上述生理参数的响应。研究结果显示, 与达里湖未洄游的瓦氏雅罗鱼相比, 洄游到贡格尔河后其血清Na+含量显著降低(P<0.05), Cl?含量显著升高(P<0.05), 肾脏和肠组织中Na+/K+-ATPase活性显著升高(P<0.05), 而鳃组织中Na+/K+-ATPase活性无显著变化; 血清K+、Ca2+、Mg2+水平和GH、IGF-1、PRL含量无显著变化。将达里湖瓦氏雅罗鱼转入河水中24h后, 其血清Cl?含量显著升高(P<0.05)、K+含量显著降低(P<0.05), 且在鳃、肠和肾组织中Na+/K+-ATPase及鳃Ca2+/Mg2+-ATPase活性均显著升高(P<0.05), 血清PRL和IGF-1水平显著升高(P<0.05); 比较湖中和河中瓦氏雅罗鱼鳃组织形态结构, 显示湖中瓦氏雅罗鱼鳃基底膜分布着大量黏液细胞, 洄游到河水中后黏液细胞数量明显减少, 鳃基底膜上氯细胞体积增大而数量未见明显变化。本研究结果表明: 瓦氏雅罗鱼从达里湖洄游到贡格尔河后通过提高血清PRL和IGF-1水平, 进而介导鳃、肠和肾组织中Na+/K+-ATPase活性增加, 从而维持鱼体较高或稳定的血清离子水平。     

糖尿病大鼠的SOD活性与抗氧化剂治疗对机体抗氧化状态的影响

目的探究糖尿病大鼠主要脏器SOD活性及蛋白表达水平的变化,并观察抗氧化剂N-乙酰半胱氨酸(N-acetylcysteine,NAC)短期治疗(4周)后对机体抗氧化状态的影响。方法 STZ诱导的糖尿病大鼠(D组,n=8)每天给予NAC 1.5 g/kg灌胃治疗(D+N组,n=8),正常对照组(C组,n=8)同时给予同体积生理盐水。4周后,获取心、肺、肝、肾组织,试剂盒检测血浆总SOD、总抗氧化物浓度、脂质过氧化特异性标志物15-F2t-isoprostane及各组织总超氧化物歧化酶(superoxide dismutase,SOD)活性,Western blotting分析SOD亚型Cu/Zn-SOD及MnSOD蛋白表达水平。结果与C组相比,D组大鼠血浆15-F2t-isoprostane与总抗氧化物浓度及心肌组织中总SOD活性显著升高,而血浆、肺、肝、肾组织总SOD活性显著降低;心、肺组织中Cu/Zn-SOD蛋白表达水平明显升高,而肝、肾组织中明显降低;肺、肾组织中Mn-SOD蛋白表达水平明显降低,而肝组织明显升高,但心肌组织变化不明显。NAC干预能不同程度逆转上述改变,但进一步降低肾组织Mn-SOD表达。结论糖尿病大鼠各组织中总SOD活性、Cu/Zn-SOD及Mn-SOD蛋白表达水平具有组织差异性,抗氧化剂NAC能不同程度恢复糖尿病大鼠各组织抗氧化水平,从而起到阻止或延缓糖尿病相关的靶器官功能损害的作用。     

烧伤患者血浆IGFBP-1水平的变化及临床意义

目的:探究烧伤患者血浆胰岛素样生长因子结合蛋白-1(IGFBP-1)水平的变化及其临床意义。方法:选择2013年1月-2015年1月我院收治的70例烧伤患者作为研究组,依据总体表烧伤面积(TBSA)将患者分为4个亚组:Ⅰ组(1%~15%)34例、Ⅱ组(16%~30%)9例、Ⅲ组(31%~50%)17例、Ⅳ组(51%~95%)10例;选取同期于我院体检健康者50例作为对照组。测定研究者IGFBP-1、胰岛素样生长因子(IGF-1)水平,分析其与TBSA的相关性。结果:在10d观察期内,Ⅲ组和Ⅳ组患者的IGF-1水平均显著低于对照组(P0.05),而Ⅱ组患者IGF-1水平仅在烧伤后1 d时与对照组存在差异(P0.05),且在烧伤后2 d,随着分级的增加,血浆IGF-1水平有下降的趋势,组间差异具有统计学意义(P0.05);研究组患者的IGFBP-1水平均显著高于对照组(P0.05);随着分级的增加,血浆IGFBP-1有上升的趋势,组间差异具有统计学意义(P0.05);线性相关性分析显示,烧伤患者血浆IGF-1水平与TBSA值呈负相关(r=-0.693,P0.05);血浆IGFBP-1水平与TBSA值呈正相关(r=0.377,P0.05)。结论:烧伤患者血浆IGF-1水平显著下降,而IGFBP-1水平则显著升高,与烧伤严重程度密切相关,二者可作为判断烧伤患者预后的重要预警指标。     

新生儿窒息与血管内皮生长因子和胰岛素样生长因子-1的关系

目的:探讨血管内皮生长因子(VEGF)及胰岛素样生长因子-1(IGF-1)在窒息新生儿脐血水平变化的临床意义.方法:采用双抗体夹心酶联免疫吸附法(ELISA)及放射免疫分析法(RIA)测定正常新生儿(67例,对照组)和窒息新生儿(50例,窒息组)脐血血管内皮生长因子(VEGF)及胰岛素样生长因子-1(IGF-1)的水平.结果:新生儿窒息时脐血VEGF比正常脐血对照组显著升高(t=-9.944,p<0.01),IGF-1比正常脐血对照组显著下降(t=-15.943,p<0.01).结论:窒息新生儿VEGF水平的上升与IGF-1水平的下降,提示两者均可能参与新生儿窒息的病理生理过程,同时检测新生儿脐血VEGF与IGF-1可望成为反映新生儿窒息程度的一个敏感和特异的指标.     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Cytoskeleton and mitochondrial morphology and function

It has been well established that the cytoskeleton is an essential modulator of cell morphology and motility, intracytoplasmic transport and mitosis, however cytoskeletal linkage to the organelles has not been unequivocally demonstrated. Indeed, cytoskeleton appears to be essential in determining and modulating gene phenotype as a function of cellular environment. According to recent studies, the organization of the cytoskeleton network together with associated protein(s) could be essential in regulating mitochondrial function and particularly the permeability of the mitochondrial outer membrane to ADP. The aim of this chapter is to summarize the main properties of the cytoskeletal environment of mitochondria and the possible role(s) of this network in mitochondrial function in myocytes.     

人血管能抑素基因的克隆、表达及其表达产物的纯化和生物活性测定

以人胎盘脐带组织为材料,提取组织总RNA,用netRTPCR方法合成人血管能抑素cDNA基因,将该cDNA克隆进pSP72载体获得重组质粒pSP72C, DNA序列分析结果与预期序列一致。用BamHⅠ和NdeⅠ双酶切,切下pSP72C上的血管能抑素cDNA,插入pET3c载体的相应位点获得重组表达质粒pETC, 转化E. coli BL21(DE3), SDSPAGE分析显示：在IPTG诱导下,血管能抑素基因获得了高效表达,表达量约占菌体总蛋白的 27.9 %,主要以包涵体形式存在。包涵体经过洗涤、裂解、蛋白复性以及Sephadex G75凝胶过滤层析等步骤后,获得了纯度达91.4 %的人血管能抑素。CAM实验证明10 μg纯化蛋白就能显著抑制鸡胚新生血管生成。