维生素C和E混合饲喂对中华鳖幼鳖抗酸应激能力的影响

来自甲鱼养殖场的60只中华鳖(Pelodiscus sinensis)幼鳖驯养3周后,实验设5组：对照组、处理Ⅰ、Ⅱ、Ⅲ和Ⅳ组。各组设2个平行,依次在饵料中混合添加维生素C(Vc)和E(VE)为0和0、250和50、2500和50、250和250、2500和250mg／kg,喂食4周后,每组取半数幼鳖经酸应激处理24h。取幼鳖血液,用镜检法测定血细胞的吞噬率,透射比浊法测定血清溶菌活力、杀菌活力以及补体C3和C4含量。①经酸应激与未经酸应激处理相比：对照组血细胞吞噬率显著降低,而处理Ⅰ-Ⅳ组无显著变化;对照组和处理Ⅰ组血清溶菌活力和补体C3含量显著下降,而处理Ⅱ-Ⅳ组无显著变化;血清杀菌活力均有显著下降(对照组、处理Ⅰ和Ⅲ组极显著,处理Ⅱ组和Ⅳ组显著);对照组、处理Ⅰ和Ⅲ组血清补体C4显著下降,而处理Ⅱ和Ⅴ组无显著变化。②经酸应激处理,血细胞吞噬率、血清溶菌活力、杀菌活力和补体C3含量,处理Ⅰ～Ⅳ组的均显著高于对照组,处理Ⅳ组显著高于其他4组;血清杀菌活力,处理Ⅱ组又高于处理Ⅰ和Ⅲ组;血清补体C4,对照组显著低于处理Ⅰ-Ⅳ组,而处理Ⅰ-Ⅳ组间无显著相异。Vc和VE混合饲喂对酸应激后中华鳖血细胞吞噬率、血清溶菌活力、杀菌活力和补体C3含量有显著协同促进作用,对血清补体C4的合成无协同作用。说明Vc和VE混合饲喂能显著增强中华鳖抗酸应激能力,缓解或部分缓解酸应激造成的不利影响。     

黄芪和酸应激对中华鳖幼鳖血清补体C3和C4含量的影响

首次探讨了黄芪和酸应激对中华鳖（Pelodiscus sinensis)幼鳖血清补体C3和C4含量的影响。实验设对照组和实验组,实验组在饵料中按5％添加黄芪原料粉。持续饲喂中华鳖幼鳖4周后取一半样,其余作酸处理24h后再取样,测定补体C3和C4的含量。结果表明,黄芪对补体C3和C4的合成有明显的促进作用;酸应激可导致血清补体C3和C4的含量下降,而黄芪能抵抗酸应激所致的下降。提示黄芪具有抗酸应激的作用。     

维生素C和E合用对应激和非应激中华鳖幼鳖生长、肝脏维生素C和E以及血清皮质醇含量的影响

为探讨维生素C (VC)和维生素E (VE)联用对应激和非应激中华鳖幼鳖的生长、肝脏VC 和VE 以及血清皮质醇含量的影响 ,作者使用了 5组饵料 ,VC 和VE 的添加量依次为 0和 0mg/kg (对照组 )、 2 5 0和 5 0mg/kg (实验Ⅰ组 )、2 5 0 0和 5 0mg/kg (实验Ⅱ组 ) ;2 5 0和 2 5 0mg/kg (实验Ⅲ组 ) ;2 5 0 0和 2 5 0mg/kg(实验Ⅳ组 )。中华鳖幼鳖的生长、肝脏VC 和VE以及血清皮质醇分别通过特定生长率、高压液相色谱法和放免法来测定。结果实验Ⅰ -Ⅳ组中华鳖的特定生长率明显高于不加VC 和VE的对照组 ,但实验Ⅰ -Ⅳ组间没有明显不同。非应激中华鳖肝脏VC 和VE的含量随饵料中VC 和VE 含量的增加而明显升高 ,并且实验Ⅱ -Ⅳ组肝脏VC 和VE都明显高于对照组和实验Ⅰ组。酸应激后 ,对照组和实验组中华鳖肝脏VC 和VE都有下降的趋势 ,但无显著差异 ;应激后实验Ⅱ -Ⅳ组肝脏VC 和VE均明显高于对照组 ,实验Ⅳ组明显高于其它 4组。血清皮质醇的含量在实验Ⅰ -Ⅳ组间没有明显不同 ,实验Ⅰ -Ⅲ组与对照组相比虽有降低的趋势但没有变化 ,实验Ⅳ组则明显低于对照组。酸应激后 ,对照组血清皮质醇明显升高 ,其他 4组虽有升高的趋势 ,但没有明显变化。应激后实验Ⅰ -Ⅳ组血清皮质醇的含量均明显低于对照组 ,实验Ⅰ -Ⅳ组间没有明显     

维生素C和E合用对中华鳖幼鳖非特异性免疫功能的影响

本研究旨在探讨维生素C和E合用对中华鳖幼鳖非特异性免疫功能的影响。实验设 5组 ,5组饵料中维生素C和E的添加量依次为 0和 0mg/kg(对照组 ) ;2 5 0和 5 0mg/kg(实验Ⅰ组 ) ;2 5 0 0和 2 5 0mg/kg(实验Ⅱ组 ) ;2 5 0和2 5 0mg/kg(实验Ⅲ组 ) ;2 5 0 0和 2 5 0mg/kg(实验Ⅳ组 )。喂食 4周 ,取其血液 ,测定中华鳖血细胞的吞噬率、血清溶菌活力、杀菌活力、补体C3和C4。结果表明 ,实验Ⅱ—Ⅳ组 ,血细胞吞噬率和血清杀菌活力明显高于对照组 ,实验Ⅳ组明显高于其他 4组 ,并且维生素C和E之间有明显的协同作用。血清溶菌活力和补体C3的含量 ,在实验Ⅰ—Ⅳ组明显高于对照组 ,实验Ⅳ组明显高于其他 4组。补体C4在实验Ⅰ—Ⅳ组明显高于对照组 ,但实验Ⅰ—Ⅳ组间没有明显差别 ,维生素C和E间也没有协同作用。     

维生素C和酸应激对中华鳖幼鳖血清补体C3和C4含量的影响

为研究维生素C对中华鳖(Pelodiscus sinensis)血清补体C3和C4的影响及其在酸应激条件下的变化,我们设置了6个实验组,饵料中维生素C的添加量依次为0、250、500、2500、5000和10000mg／kg,喂食4周后取其血清,用透射比浊法测定酸应激前后中华鳖血清补体C3和C4的含量。结果表明,维生素C添加量为250mg／kg时,血清补体C3的含量与对照组间没有明显不同;维生素C添加量为500、2500、5000和10000mg／kg的4组,血清补体C3的含量明显高于对照组和维生素C添加量为250mg／kg组;维生素C添加量为500mg／kg的一组,血清补体CA含量明显高于其它5组;维生素C添加量为250mg／kg组明显高于10000mg／kg组。酸应激后,补体C3的含量没有明显下降,将维生素C添加量为0、250和500mg／kg的三组并为一组处理,则应激后有明显下降。维生素C添加量为0、250和500mg／kg的3组,血清补体CA的含量在酸应激后明显下降,而维生素C添加量为2500、5000和10000mg／kg的3组,应激后血清补体C4没有明显变化。维生素C和酸应激对中华鳖血清补体C3和CA含量的影响没有交互作用。这说明,维生素C在一定剂量范围内,能提高中华鳖血清补体C3和CA的水平,酸应激能导致其含量降低,而高剂量的维生素C对其下降有颉颃作用[动物学报49(6)：769～774,2003]。     

维生素C多聚磷酸酯对小鼠抗热应激能力的影响

为研究维生素C多聚磷酸酯对小鼠抗热应激能力的影响 ,将 4 8只 3～ 4周龄、体重为 16～ 2 9g的健康雄性小鼠随机分为 4组 (对照组、Ⅰ、Ⅱ和Ⅲ组 ) ,在其饵料中分别添加 0、 5 0 0、 2 5 0 0和 5 0 0 0mg/kg的35 %维生素C多聚磷酸酯。喂食 4周后 ,每组取一半小鼠处死取其肝脏 ,另一半置于 (35± 1)℃条件下 ,2 4h后作同样处理。用硫代巴比妥酸分光光度法测肝脏脂质过氧化物 (LPO)的含量 ,用亚硝酸盐形成法测超氧化物歧化酶 (SOD)的活性 ,用分光光度法测过氧化氢酶 (CAT)和谷胱甘肽过氧化物酶 (GSH Px)的活性。经热应激与未经热应激相比 ,LPO :对照组和Ⅰ组显著升高 ,而Ⅱ和Ⅲ组无显著差异。SOD和GSH Px :对照组显著下降 ,其他 3组无显著差异 ;其中SOD :Ⅱ和Ⅲ组显著高于对照组和Ⅰ组 ,GSH Px :Ⅲ组显著高于其他 3组。CAT :对照组和Ⅰ组显著降低 ,而Ⅱ和Ⅲ组无显著差异 ;Ⅱ和Ⅲ组显著高于对照组和Ⅰ组。表明热应激促进了小鼠肝脏LPO的产生 ,抑制了抗化物酶的活性 ;而维生素C多聚磷酸酯对热应激造成的不利影响有缓解作用。     

维生素C多聚磷酸酯对小鼠生长、淋巴细胞转化以及血清皮质酮含量的影响

目的探讨维生素C多聚磷酸酯对小鼠生长、淋巴细胞转化及血清皮质酮含量的影响以及淋巴细胞转化和血清皮质酮含量在热应激条件下的变化。方法在4组小鼠的饲料中依次添加35%的维生素C多聚磷酸酯,添加剂量分别为0(对照组)、500(实验Ⅰ组)、2500(实验Ⅱ组)和5000mg/kg(实验Ⅲ组),在(21±1)℃下饲养4周,对照组和实验Ⅱ组各取一半小鼠经(35±1)℃热应激处理24h。用特定生长率的方法测定小鼠的生长,用噻唑蓝还原法测定淋巴细胞转化,用荧光法测定血清中皮质酮的含量。结果各组间ICR小鼠的生长、淋巴细胞转化无明显差异;实验Ⅱ组的血清皮质酮含量明显低于对照组,其他组间差异无显著性。热应激后,淋巴细胞转化无明显变化;血清皮质酮含量较应激前有显著上升,但实验Ⅱ组仍显著低于对照组。结论在饲料中添加维生素C多聚磷酸酯对小鼠的特定生长率和淋巴细胞转化均无显著影响,但会降低小鼠血清皮质酮的含量。     

维生素E对中华鳖幼鳖生长、肝脏维生素E以及血清皮质醇含量的影响

为探讨维生素E(VE)对中华鳖(Pelodiscus sinensis)幼鳖的生长、肝脏VE和血清皮质醇的影响，通过特定生长率、高压液相色谱法和放免法，我们测定了中华鳖幼鳖的生长、肝脏VE和血清皮质醇含量。发现VE添加量为1000和5000mg／kg的两组，能明显降低中华鳖幼鳖的生长。维生素E添加量为500、1000和5000mg／kg的三组，肝脏维生素E含量明显高于对照组，VE添加量在0—1000mg／kg的范围时，肝脏VE的含量随着饲料中VE含量的增加呈指数式增加，并且在VE添加量为5000kg／kg的一组基本达到饱和。维生素E添加量为0和50mg／kg的2组，其血清皮质醇的平均值明显高于维生素E添加量为250、500、1000和5000mg／kg的4组的平均值。上述结果表明：高剂量的VE降低了中华鳖幼鳖的生长和血清皮质醇的含量；在一定剂量范围内，肝脏VE随着饲料中VE含量的增加而升高。     

不同C/N生物絮团对洛氏鱥急性铜暴露保护作用

实验旨在探索不同C/N生物絮团对急性铜暴露洛氏鱥免疫抑制、炎症反应与氧化应激的保护作用。挑选480尾洛氏鱥(Rhynchocypris lagowskii Dybowski)幼鱼(10±0.15) g, 随机分成4组, 每组3个重复, 每个重复40尾鱼。对照组饲喂商品料(C/N 10.8﹕1), 实验组选择葡萄糖为外添碳源调节C/N 15﹕1(Ⅱ组)、C/N 20﹕1(Ⅲ组)和C/N 25﹕1(Ⅳ组), 生长实验为56d, 之后进行为期96h的急性铜暴露胁迫实验。结果表明, 各实验组中免疫、抗氧化酶活性与炎症因子含量随着C/N的增加先升高后下降。其中, 与对照组相比, Ⅲ和Ⅳ组血清碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、溶菌酶(LSZ)、一氧化氮合酶(NOS)、补体C3、C4和免疫球蛋白M(IgM)水平显著升高(P<0.05), 而Ⅲ和Ⅳ组之间补体C3和C4水平差异不显著(P>0.05); Ⅲ和Ⅳ组血清过氧化氢酶(CAT)、抗坏血酸(ASA)、超氧化物歧化酶(SOD)、总抗氧化能力(T-AOC)、谷胱甘肽过氧化物酶(GSH-PX)与谷胱甘肽还原酶(GR)酶活性显著高于对照组(P<0.05), 而丙二醛(MDA)含量显著低于对照组(P<0.05)。同时Ⅱ组的SOD和CAT酶活性也有所升高, 且与对照组差异显著(P<0.05); Ⅲ和Ⅳ组血清肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)与白介素-6(IL-6)含量显著低于对照组(P<0.05), 且它们两组之间无显著差异(P>0.05)。然而, Ⅱ、Ⅲ和Ⅳ组白介素-2(IL-2)含量均显著高于对照组, 且第Ⅲ组含量最高。在实验条件下, 当C/N≥15: 1时, 生物絮团能有效地增强急性铜暴露下洛氏鱥的免疫与抗氧化酶活力, 且在C/N为20﹕1时效果最为显著。     

过敏性紫癜合并肺炎支原体感染患儿呼吸道菌群多样性与补体C3、C4及免疫指标的相关性

目的研究儿童过敏性紫癜(HSP)合并肺炎支原体(MP)感染患儿呼吸道菌群多样性与补体C3、C4及免疫指标的相关性。方法选择2017年3月至2020年3月于我院接受治疗的HSP患儿96例,其中48例合并MP感染的患儿设为合并MP组,48例未合并MP感染的患儿设为HSP组。另外选择同期我院健康体检儿童45例为对照组。比较3组对象呼吸道菌群多样性,血清免疫球蛋白IgG、IgA、IgM、IgE及补体C3、C4水平,并分析呼吸道菌群多样性与免疫指标间的相关性。结果合并MP组患儿呼吸道菌群Simpson指数显著高于对照组,Shannon指数显著小于对照组(均P<0.05)。HSP组患儿呼吸道菌群Simpson指数及Shannon指数与对照比较差异无统计学意义(均P>0.05)。合并MP组患儿血清IgA水平较对照组显著上升,IgM水平显著下降;HSP组患儿血清IgA、IgE水平较对照组显著上升;合并MP组患儿血清IgE、IgM水平显著低于HSP组(均P<0.05);而3组对象血清IgG水平比较差异无统计学意义(均P>0.05)。合并MP组和HSP组患儿血清补体C3水平显著低于对照组;合并MP组患儿血清补体C4水平显著低于对照组;合并MP组患儿血清补体C3、C4水平均显著低于HSP组(均P<0.05)。IgM、IgE及补体、C3、C4水平与Simpson指数呈负相关,与Shannon指数呈正相关(均P<0.05)。结论HSP合并MP感染患儿呼吸道菌群多样性与补体C3、C4及IgM、IgE水平具有显著相关性。     

Complement C3 opsonization of Chlamydia trachomatis facilitates uptake in human monocytes

Chlamydia trachomatis is an obligate intracellular bacterium that causes severe infections, which can lead to infertility and ectopic pregnancy. Although both innate and adaptive immune responses are elicited during chlamydial infection the bacterium succeeds to evade host defense mechanisms establishing chronic infections. Thus, studying the host–pathogen interaction during chlamydial infection is of importance to understand how C. trachomatis can cause chronic infections. Both the complement system and monocytes play essential roles in anti-bacterial defense, and, therefore, we investigated the interaction between the complement system and the human pathogens C. trachomatis D and L2.Complement competent serum facilitated rapid uptake of both chlamydial serovars into monocytes. Using immunoelectron microscopy, we showed that products of complement C3 were loosely deposited on the bacterial surface in complement competent serum and further characterization demonstrated that the deposited C3 product was the opsonin iC3b. Using C3-depleted serum we confirmed that complement C3 facilitates rapid uptake of chlamydiae into monocytes in complement competent serum. Complement facilitated uptake did not influence intracellular survival of C. trachomatis or C. trachomatis-induced cytokine secretion. Hence, C. trachomatis D and L2 activate the complement system leading to chlamydial opsonization by iC3b and subsequent phagocytosis, activation and bacterial elimination by human monocytes.     

Mapping of the complement C9 binding domain in paramyosin of the blood fluke Schistosoma mansoni

Schistosomes are believed to evade complement-mediated damage by expression of complement inhibitory proteins. Our previous results [Deng, J., Gold, D., LoVerde, P.T., Fishelson, Z., 2003. Inhibition of the complement membrane attack complex by Schistosoma mansoni paramyosin. Infect. Immun. 71, 6402-6410.] have demonstrated that paramyosin (Pmy) of the blood fluke S. mansoni binds to the human complement proteins C8 and C9, inhibits complement activation at the terminal stage and protects the parasite from complement-mediated damage. In order to locate the Pmy binding site to C8 and C9, various fragments of Pmy cDNA were PCR-cloned into a pET28a bacterial expression vector. Recombinant His-tagged Pmy fragments were expressed in BL21 Escherichia coli and purified over a nickel-nitrilotriacetic acid column. Binding assays by Western blotting with monoclonal anti-His antibody demonstrated that PmyCC (Pmy amino acids (744)Asp-(866)Met) was the only Pmy fragment that bound to human C8 and C9. Functional analyses demonstrated that PmyCC inhibited hemolysis of rabbit erythrocytes and of antibody-sensitized sheep erythrocytes by human complement. Importantly, PmyCC inhibited in vitro killing of trypsin-sensitized schistosomula of S. mansoni by human complement. In the presence of PmyCC, Zn(2+)-induced C9 polymerization was inhibited. Most of the immunodominant B-cell antigenic epitopes of Pmy are present in the PmyCC region, as antibodies collected from mice immunized with recombinant Pmy bound primarily to PmyCC. Taken together, this study has mapped the complement regulatory domain in Pmy, capable of binding to C8 and C9 and preventing polyC9 formation, to its C-terminal region.     

中国湖北地区汉族家系补体第四成分（C4）单倍型的检测

用我室仿国际标准C4定型程序改进后建立的方法及羧肽酶B处理后C4分型方法对湖北地区93个无血缘关系的汉族家系进行C4单倍型的检测,对310个C4单倍型分析可见,我国汉族以A3B1频率最高(0.4194),A3B2次之(0.1161),A2B1与A4B2均为0.0903。以下依次是AQOB1(0.0645)、A3BQO(0.0548)、AQOBQO(0.0322)、A2B2(0.0256)、A4B1(0.0161)、A2B92(0.0129)等。从连锁不平衡参数(Δ)的卡方数值可见,A4B2、AQOBQO及A2B92具有极显著意义的阳性△值;而A4B1与AQOB2则具有极显著意义的阴性△值。将我们的结果与日本人、美国及德国白人,南非黑人的资料进行了对比,并进行了一些讨论。     

Adiponectin binds C1q and activates the classical pathway of complement

The adipose-specific protein adiponectin binds to a number of target molecules, including damaged endothelium and the surface of apoptotic cells. However, the significance of this binding remains unclear. This study demonstrates the binding of purified C1q to recombinant adiponectin under physiological conditions, and the dependence of this upon Ca⁺⁺ and Mg⁺⁺. Binding was enhanced by metaperiodate-mediated destruction of glucosylgalactosyl sugars on adiponectin. Adiponectin was bound by the globular domain of the A chain of collagenase-digested C1q, and C1q binding induced deposition of C4 and C3 through activation of the classical complement pathway. After Western blotting, affinity-purified adiponectin from human serum bound C1q, whereas adiponectin in whole serum did not, unless pre-treated with metaperiodate. These results suggest adiponectin is member of the pattern-recognition family of defence collagens, able to bind target molecules and activate complement. It may therefore play an important role in innate immunity and autoimmune phenomena.     

Complete sequencing and expression of three complement components,C1r,C4 and C1 inhibitor,of the classical activation pathway of the complement system in rainbow trout<Emphasis Type="Italic"> Oncorhynchus mykiss</Emphasis>

Three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway have been fully sequenced and their expression investigated in rainbow trout (Oncorhynchus mykiss). Trout C1r cDNA encodes a 707-amino-acid (aa) protein with a theoretical M _r of 77,200. The trout translation shows highest homology with carp C1r/s, and lower, equal homologies to mammalian C1r and C1s, and MASPs from other vertebrate species. However, phylogenetic analysis and structural features suggest that the trout sequence, together with the two carp sequences, are the orthologues of mammalian C1r. The trout C4 cDNA encodes a 1,724-aa protein with a theoretical M _r of 192,600. The trout translation shows higher homologies to the carp C4B and medaka C4, but lower homologies to C4 from other species and the carp C4A. It has a predicted signal peptide of 22 aa, a -chain of 773 aa, a -chain of 635 aa and a -chain of 288 aa. Trout C1 inhibitor cDNA encodes a 611-aa protein with a theoretical M _r of 68,700. The trout translation has a C-terminal serpin domain with high homologies with mammalian counterparts (~37% identities), and a longer N-terminus, with no significant homology to other serpins, which contains two Ig-like domains. A molecule containing two Ig-like domains followed by a serpin domain, has also been found in an EST clone from another bony fish, the Japanese flounder. This suggests a unique structural feature of C1 inhibitor in fish. The functional significance of the Ig domains is discussed. The liver is the major site of expression of the three trout complement components, C1r, C4 and C1 inhibitor, although their expression is also detectable in other tissues. The extra-hepatic expression of complement genes may be important for local protection and inflammatory responses. Low-level constitutive expression of the three components was also detectable in a trout monocyte/macrophage cell line RTS-11, but only the expression of C4 could be upregulated by LPS.The nucleotide sequence data will appear in the EMBL/DDBJ/GenBank nucleotide sequence database under the following accession numbers: AJ519929 (trout C1r), AJ519930 (trout C1 inhibitor), AJ544262 (trout C4) and BN000290 (flounder C1 inhibitor)     

Design and optimization of aniline-substituted tetrahydroquinoline C5a receptor antagonists

A series of aniline-substituted tetrahydroquinoline C5a receptor antagonists were discovered. A functionality requirement of ortho substitution on the aniline was revealed. Secondary anilines, in general, outperformed tertiary analogs in inhibition of C5a-induced calcium mobilization. Further enhancement of activity was realized in the presence of an ortho hydroxyalkyl side chain. The functional IC₅₀ of selected analogs was optimized to the single-digit nanomolar level.     

Effects of zinc on factor I cofactor activity of C4b-binding protein and factor H

Complement inhibition is to a large extent achieved by proteolytic degradation of activated complement factors C3b and C4b by factor I (FI). This reaction requires a cofactor protein that binds C3b/C4b. We found that the cofactor activity of C4b-binding protein towards C4b/C3b and factor H towards C3b increase at micromolar concentrations of Zn(2+) and are abolished at 2 mM Zn(2+) and above. 65Zn(2+) bound to C3b and C4b molecules but not the cofactors or FI when they were immobilized in a native form on a nitrocellulose membrane. Zn(2+) binding constants for C3met (0.2 microM) and C4met (0.1 microM) were determined using fluorescent chelator. It appears that higher cofactor activity at low zinc concentrations is due to an increase of affinity between C4b/C3b and cofactor proteins as assessed by surface plasmon resonance. Inhibition of the reaction seen at higher concentrations is due to aggregation of C4b/C3b.