复合益生菌制剂“海生元”治疗实验性肠菌群失调症疗效机制探讨

目的 :通过观察复合益生菌制剂“海生元”各重要组分对实验性肠菌群失调症的治疗效果 ,初步探讨“海生元”纠正肠菌群失调症的疗效机制。方法 :(1)制备SPF BALB/c小鼠肠菌群失调症模型 ;(2 )处理因素分组 :对照组设自然恢复组 (空白对照 )和生理盐水组 (安慰剂对照 )。治疗组按“海生元”制剂中的主要药效学干预成分 (组成成分 )分为 :T9菌液 (双歧杆菌 )组 ,M9菌液 (乳杆菌 )组 ,螺旋藻组 ,组方中药组 ,复方制剂 (“海生元”)组。 (3)比较观察各组疗后第 2、4、6天腹泻症状恢复情况及全程治疗后 (共 6d)肠菌群定量分析结果。结果 :(1)各处理组对模型小鼠腹泻恢复的影响 :疗前各组 10 0 %为Ⅱ～Ⅲ (中～重 )度腹泻。疗后第 2天 ,两对照组Ⅱ～Ⅲ度腹泻为 99%和 93% ,而 5个治疗组依次为 5 3%、5 1%、5 8%、6 7%和4 7% ;疗后第 4天 ,两对照组Ⅱ～Ⅲ度腹泻为 6 5 %和 5 7% ,而 5个治疗组依次为 15 %、2 1%、19%、2 4 %和16 % ;疗后第 6天 ,两对照组和组方中药组Ⅱ度腹泻率分别为 36 %、2 9%和 18% ,其余各组均正常或为I度(轻度 )腹泻。上述 7个处理组大便正常率分别为 2 1%、2 1%、86 %、79%、77%、4 5 %和 89% ,以复方制剂(“海生元”)组最高。 (2 )各处理组肠菌群定量分析 :两对照组疗后第 2、4、6天 。

复合益生菌制剂“海生元”对实验性肠菌群失调症调整作用的研究

目的 :观察及评价复合益生菌制剂“海生元”对实验性肠菌群失调症的纠正作用。方法 :1)制备复合益生菌制剂“海生元” ;2 )制备SPF·BALB/c小鼠肠菌群失调症模型 ;3)观察“海生元”对模型小鼠 (复方制剂 )组灌胃治疗效果 ;设自然恢复对照组和生理盐水对照组 (安慰组 ) ;动态比较疗后第 2、4、6天各组腹泻症状恢复情况和肠菌群定量分析。结果 :1)小鼠肠菌群失调症造模效果分析 :氨苄青霉素造模前 ,10 0 %小鼠为正常大便 (无腹泻 ) ;造模第 3天 ,6 0 %为重度 (Ⅲ度 )腹泻 ,4 0 %为中度 (Ⅱ度 ) ,即 10 0 %模型小鼠出现中、重度腹泻 ,表明造模成功。此外 ,模型小鼠肠菌群中双歧杆菌、乳杆菌未检出 ,肠杆、肠球、类杆菌数量大大低于正常小鼠 (P <0 0 1)证实出现了菌群失调症。 2 )各组腹泻恢复情况 :疗前各组均为中、重度腹泻。疗后第 2天 ,复方制剂组Ⅱ～Ⅲ度腹泻降至 4 7% ,而自然恢复组和生理盐水组分别为 99%和93% ;疗后第 4天 ,复方制剂组Ⅱ～Ⅲ度仅 16 % ,两个对照组则分别为 6 5 %和 5 7% ;疗后第 6天 ,复方制剂组完全正常者为 89% ,而两对照组分别为 2 1%。 3)各组肠菌群动态定量分析 :自然恢复和生理盐水两个对照组 ,造模后第 2、4、6天五种肠道正常生理菌数量逐渐递升 。

复合益生菌制剂“海生元”的卫生安全性检验

目的 :监测和评估复合益生菌制剂“海生元”在制备、保存和使用过程中微生物污染状况 ,消除对健康造成潜在不良影响的危险。方法 :在有效期 (1年 )和保存 2 4个月 (2年 )时 ,定期随机对“海生元”口服液和胶囊取样 ,按常规方法进行微生物限度检验 ,包括细菌总数、大肠菌群、致病菌检查及霉菌、酵母菌计数。结果 :(1)贮存 12个月时微生物限度检验结果显示 :口服液各项指标均为阴性。胶囊剂分别为细菌数 <2× 10CFU/ g ,大肠菌群数 <3MPN/ 10 0 g、真菌和酵母菌总数分别 <5CFU/g ,未检出致病菌。 (2 )储存 2 4个月时的微生物限度检验结果显示 :口服液细菌总数、霉菌数和酵母菌数均 <1CFU/ g ,大肠菌数 <3MPN/ 10 0 g。胶囊剂细菌总数 <4× 10 3 CFU/ g ,大肠菌数 <30MPN/ 10 0g ,霉菌和酵母菌数 <10CFU/ g ,未检出致病菌。结论 :复合益生菌制剂“海生元”2种剂型 (口服液和胶囊 )在效期内的染菌情况均符合规定限度要求。在普通条件下储存 2 4个月 (2年 ) ,仅对胶囊的细菌总数略有影响 ,但结果仍符合中国药典规定的限度标准。本实验结果证实了复合益生菌制剂制备、储存过程中卫生安全的可靠性。

复合益生菌制剂“海生元”制备各阶段效力稳定性观察

目的 :检测复合益生菌制剂“海生元”制备储存各阶段益生菌的存活数量及其生物表型的稳定性 ,以判定该制剂的效力稳定性。方法 :在有效期内 (1年 ) ,每月定期随机取 5个“海生元”胶囊 ,按常规对M9(乳杆菌QJ4 0 5 )和T9(双歧杆菌QJ4 0 5 )两株益生菌进行活菌培养计数和菌种鉴定。结果 :(1)在常温保存条件下的一年效期内 ,制剂中的 2种益生菌 (M9和T9)存活数量略有下降趋势 ,但各次结果均符合产品的效力要求 (活菌数 >10 8/ g)。 (2 )M9和T9各次纯培养的形态学、生化反应模式及终末代谢产物气相色谱定量分析结果均分别符合其乳杆菌和双歧杆菌的菌种鉴定特征。结论 :复合益生菌制剂“海生元”在常温保存的 1年有效期内 ,可保持制剂中益生菌的活菌数量及稳定的生物表型特征 ,从而保证该制剂的效力稳定性和疗效要求。

复合益生菌制剂“海生元”急性毒性试验研究

目的:观察复合益生菌制剂"海生元"(HSY)及其核心成分-益生菌(M9、T9)经两种途径给药的急性毒性反应,评价其安全性.方法:以健康小鼠为对象,将HSY、M9、T9三种受试物按i.p(腹腔注射)和p.o(口服灌胃)两种给药方式进行急毒试验.每种给药方法分5个剂量组.i.p小鼠用Bliss法测得受试物的LD50和95%可信限,p.o小鼠用安全限度试验测得受试物的最大耐受量.结果:1)i.p法测得各受试物LD50和95%可信限:M9为898.31 mg/ml(770.18～925.79);T9为1 856.95 mg/ml(1 521.48～2 010.36);HSY为7 605.23 mg/ml(6 980.52～8 304.11).2)p.o法测得各受试物最大耐受量及耐受倍数:健康小鼠最大耐受量分别为M9>4 000 mg/ml,T9>6 000 mg/ml,HSY>15 000 mg/ml;最大耐受量倍数(即耐受成人每日用量的倍数)分别为133倍、200倍和500倍.结论: HSY、M9、T9口服给药均非常安全.三种受试物比较,又以HSY安全程度最高.

复合益生菌制剂“海生元”长期毒性试验研究

目的:观察及评价复合益生菌制剂"海生元"(HSY)的长期用药安全性.方法:以正常健康Wistar大鼠为受试对象,HSY及其核心成分M9、T9菌株为受试物,改良TPY和生理盐水(NS)为对照物,用M9、T9、HSY三种受试物的最大耐受剂量(分别为成人每日用量的125倍、200倍、500倍)口服给药4个月,给药前后及恢复期满(3周后)分别测定受试大鼠的体重、血液学及血生化指标、脏器系数并做病理学检查.结果:长期口服M9、T9及HSY,大鼠的外观、活动、食欲、大便、毛发等无明显异常变化.在给药的4个月内,除极少数指标外,三种受试物对大鼠的体重增长、外周血象、血液生化学指标、脏器系数及各组织器官的病理形态均未见有显著影响,与对照组基本一致.结论:HSY及其核心成分M9和T9菌株毒性极低,经口服长期应用是十分安全的.

Analysis on Morphological Variations among Six Wild Groups of Anthocidaris crassispina from South China Sea

In order to study the morphological variations among wild Anthocidaris crassispina groups in the southeast coast of China, six groups collected from Daya Bay, Egong Bay, Hailing Island, and Nan＇ao Island in Guangzhou Province, Weizhou Island in Guangxi Province and Ningde City in Fujian Province were analyzed in this paper. Six morphometric traits were measured and the degrees of morphological variations among wild Anthocidaris crassispina groups were compared using four multivariate analyses, one-way ANOVA analysis, variation coefficient method, cluster analysis and discriminant analysis. The results of one-way ANOVA analysis showed that there were significant differences in the six morphometric traits between Weizhou Island＇s population and the other five groups. The variation coefficient analysis showed that the difference between the Weizhou Island＇s population and Hailing Is- land＇s population reached the interspecific level. The results of cluster analysis showed that the morphometric characters of Daly Bay＇s population, Egong Bay＇s population and Nan＇ao＇s population were more similar, but the groups from Weizhou Island and Ningde City were the most divergent. The discriminant analysis showed that the identification accuracy of A. crassispina was 47.8%-86%, and the total dis- criminant accuracy was 65.3%. All the analysis results above suggest that certain morphological variation has been generated among A. crassispina groups from differ- ent origins due to geographical isolation and different environmental factors.

Benthic nutrient recycling in shallow coastal waters of the Bohai Sea

Sediment-water fluxes of N and P species in the Bohai Sea were investigated in Septem-ber-October 1998 and April-May 1999. The benthic fluxes of nutrient species were determined by incubating sediment core samples with bottom seawater bubbled with air or nitrogen. -2NO,+4NH, dissolved organic nitrogen (DON) and phosphorus (DOP), total dissolved nitrogen (TDN) and phosphorus (TDP), and -34PO showed a net exchange flux from seawater to sediment, while -3NO, dissolved inorganic nitrogen (DIN) and -23SiO were released from sediment to seawater in the Bohai Sea. Sediment-water nutrient exchange increases DIN and reduces the phosphorus load in the Bohai Sea. The release of silicate from sediment to overlying seawater reduces potential silicate limitation of primary production resulted from decrease of riverine discharge. The exchange flux of nutrients showed no obvious seasonal variation. The present study showed that the concentrations and composition of nutrients in the water column were affected by suspended sediment, and that not all the exchangeable phosphate in sediment could be released via sediment resuspension.

Influence of Dietary Probiotic Bacillus TC22 and Prebiotic Fructooligosaccharide on Growth,Immune Responses and Disease Resistance against Vibrio splendidus Infection in Sea Cucumber Apostichopus japonicus

The effects of probiotic Bacillus TC22 (isolated from intestine of infected sea cucumber) and prebiotic fructooligosaccharide (FOS) on growth, immunity and disease resistance in sea cucumber Apostichopus japonicus were studied. Six experimental diets were formulated with combinations of three levels of TC22 (0, 107 and 109 CFU g-1 diet) and two levels of FOS (0 and 0.5%) in a 3 × 2 factorial experiment. At the end of the 8-week feeding trial,animals were challenged by injecting Vibrio splendidus. The results revealed that the specific growth rates (SGR) of sea cucumbers were not affected by TC22 and FOS, or the interaction between TC22 and FOS (P > 0.05). However,there were significant interactions between TC22 and FOS for immune response and disease resistance in sea cucumbers (P < 0.05). When sea cucumbers were fed with TC22 at 109 CFU g?1 feed and 0.5% FOS alone or in combination, the phagocytosis, respiratory burst and phenoloxidase activity of sea cucumber coelomocytes were significantly enhanced;the disease resistance against V. splendidus infection was also increased significantly (P < 0.05).However, dietary combination of TC22 at 109 CFU g-1 diet and 0.5% FOS did not yield significantly higher efficiency than either TC22 at 109 CFU g-1 diet or 0.5% FOS used alone (P > 0.05). Therefore, further studies should examine the effects of combinations of other levels of FOS(> 0.5% or < 0.5%) and TC22 on the immunity and disease resistance of sea cucumbers.

Analysis of the Comprehensive Physical Field for a New Flywheel Energy Storage Motor/Generator on Ships

A novel flywheel energy storage （FES） motor/generator （M/G） was proposed for marine systems. The purpose was to improve the power quality of a marine power system （MPS） and strengthen the energy recycle. Two structures including the magnetic or non-magnetic inner-rotor were contrasted in the magnetostatic field by using finite element analysis （FEA）. By optimally designing the size parameters, the average speed of FEA results of was 17 200 r/m, and the current was controlled between 62 and 68 A in the transient field. The electrical machine electromagnetism design was further optimized by the FEA in the temperature field, to find the local overheating point under the normal operation condition and provide guidance for the cooling system. Finally, it can be concluded from the comprehensive physical field analysis that the novel redundant structure M/G can improve the efficiency of the M/G and maintain the stability of the MPS.

EphA3 functions are regulated by collaborating phosphotyrosine residues

Ephrin ligands interact with Eph receptors to regulate a wide variety of biological and pathological processes. Recent studies have identified several downstream pathways that mediate the functions of these receptors. Activation of the receptors by ephrin binding results in the phosphorylation of the receptor tyrosine residues. These phospho- rylated residues serve as docking sites for many of the downstream signaling pathways. However, the relative contributions of different phosphotyrosine residues remain undefined. In the present study, we mutated each individual tyrosine residues in the cytoplasmic domain of EphA3 receptor and studied the effects using cell migration, process retraction, and growth cone collapse assays. Stimulation of the EphA3 receptor with ephrin-A5 inhibits 293A cell mi- gration, reduces NG108-15 cell neurite outgrowth, and induces growth cone collapse in hippocampal neurons. Muta- tion of either Y602 or Y779 alone partially decreases EphA3-induced responses. Full abrogation can only be achieved with mutations of both Y602 and Y779. These observations suggest a collaborative model of different downstream pathways.

Potential Applications of X-ray Fluorescence Core Scanner in Elemental Analyses of the Muddy Sediments on the Coastal Shelves of China and in Ecological Study

Large areas of muddy sediments on the coastal shelves of China provide important samples for studying climate and ecological changes. Analysis of a large number of such samples, which is essential for systematic study on environmental information recorded in mud areas because of complicated sedimentary environment and variable sedimentary rate, requires a fast and economical method. In this study, we investigated the potential of X-ray fluorescence core scanner （XRFS）, a fast analytical instrument for measuring the elemental concentrations of muddy sediments, and observed a significant correlation between the element concentrations of muddy sediments determined by regular X-ray fluorescence spectrometer （XRF） and XRFS, respectively. The correlations are mainly determined by excitation energy of elements, but also influenced by solubility of element ions. Furthermore, we found a striking link between A1 concentrations and marine-originated organic carbon （MOC）, a proxy of marine primary productivity. This indicates that MOC is partly controlled by sedimentary characteristics. Therefore, XRFS method has a good potential in fast analysis of a large number of muddy sediment samples, and it can also be used to calibrate MOC in ecological study of coastal seas.