167 resultados para Lethal Dose 50


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This study aimed at evaluating the ploidy effects on growth performances of Chinese shrimp (Fenneropenaeus chinensis Osbeck, 1765) reared in different salinities under laboratory conditions. In the acute salinity experiment, there was no difference (P > 0.05) in tolerance observed in triploid and diploid shrimp due to abrupt salinity changes. The lethal salinity for 50% of the individuals in 96 h at 23-25 degrees C was about 2 g L-1 in both triploids and diploids. While for the chronic salinity experiment, statistical analyses confirmed that the differences in growth performances including the specific growth rate (SGR), the feeding rate (FR), feed conversion efficiency (FCE) and intermoult period (IP) between triploid and diploid were related to salinity. Diploid shrimp reared in 20 g L-1 exhibited highest SGR (P < 0.05), while triploids performed well in 20 and 30 g L-1 salinities (P < 0.05). Based on the survival and growth data, the optimal salinity for the culture of diploid F. chinensis should be 20 g L-1 and for triploids it should be between 20 and 30 g L-1.

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The high mortality rate of immunocompromised patients with fungal infections and the limited availability of highly efficacious and safe agents demand the development of new antifungal therapeutics. To rapidly discover such agents, we developed a high-throughput synergy screening (HTSS) strategy for novel microbial natural products. Specifically, a microbial natural product library was screened for hits that synergize the effect of a low dosage of ketoconazole (KTC) that alone shows little detectable fungicidal activity. Through screening of approximate to 20,000 microbial extracts, 12 hits were identified with broadspectrum antifungal activity. Seven of them showed little cytotoxicity against human hepatoma cells. Fractionation of the active extracts revealed beauvericin (BEA) as the most potent component, because it dramatically synergized KTC activity against diverse fungal pathogens by a checkerboard assay. Significantly, in our immunocompromised mouse model, combinations of BEA (0.5 mg/kg) and KTC (0.5 mg/kg) prolonged survival of the host infected with Candida parapsilosis and reduced fungal colony counts in animal organs including kidneys, lungs, and brains. Such an effect was not achieved even with the high dose of 50 mg/kg KTC. These data support synergism between BEA and KTC and thereby a prospective strategy for antifungal therapy.

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Various quaternized chitosans (QCSs) were synthesized according to previous method. Their reducing power and antioxidant potency against hydroxyl radicals ((OH)-O-center dot) and hydrogen peroxide (H2O2) were explored by the established systems in vitro. The QCSs exhibited markedly antioxidant activity, especially TCEDMCS, whose IC50 on hydroxyl radicals was 0.235 mg/mL. They showed 65-80% scavenging effect on hydrogen peroxide at a dose of 0.5 mg/mL. Generally, the antioxidant activity decreased in the order TCEDMCS > TBEDMCS > EDMCS > PDMCS > IBDMCS > Chitosan. Furthermore, the order of their (OH)-O-center dot and H2O2 scavenging activity was consistent with the electronegativity of different substituted groups in the QCSs. The QCSs showed much stronger antioxidant activity than that of chitosan may be due to the positive charge density of the nitrogen atoms in QCSs strengthened by the substituted groups. (C) 2009 Elsevier Ltd. All rights reserved.

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Effects of various concentrations of two heavy metals, namely Cd and Cu, on gametophytes of Laminaria japonica Aresch were determined by recording morphological changes of gametophytes, determining pH values and the heavy metal content of the culture solution, calculating the germination rate of sporophytes, and observing heavy metal (Cd) distribution using a fluorescence microscope. The results showed that heavy metals damaged the gametophytes, and were even lethal, and that the higher the concentration of heavy metal ions, the greater the injury to gametophytes. Gametophytes could not survive in culture solutions containing more than 100 mg/L Cd and 50 mg/L Cu and were only able to survive in culture solution containing a mixture of Cd and Cu tip to a concentration of 10 mg/L, which indicates that gametophytes have a higher tolerance to Cd than Cu and that multiple heavy metal ions in solution markedly aggravate the damage to gametophytes compared with individual heavy metal ions. With increases in the concentration of the heavy metal, the burgeoning rate of sporophytes decreased acutely, and solutions containing multiple heavy metal ions caused even more marked harm to sporophytes than solutions containing a single heavy metal ion, because most sporophytes died in mixed solutions. The pH value of the culture medium dropped immediately at the beginning (the first day) of treatment, increased over the following days, and then decreased again. The pH of culture media containing multiple heavy metal ions showed greater variation than media containing a single heavy metal ion, with the extent of the decrease in pH of culture media containing multiple ions being greatest during the last period of the experiment. With increases in the concentration of heavy metals, the capacity of gametophytes to accumulate these ions increased. The blue fluorescent light emitted by the Cd- and Cd-binding protein complex existing in gametophytes in media containing different concentrations of Cd showed clearly the distribution of the ion in gametophytes and the results obtained were consistent with distribution determined using other methods. All results of the present study showed that gametophytes of L. japonica play a remarkable role as heavy metal decontaminators, especially with regard to Cd.

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Oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs in certain contexts are known to be immunostimulatory in vertebrate systems. CpG ODNs with immune effects have been identified for many fish species but, to our knowledge, not for turbot. In this study, a turbot-effective CpG ODN, ODN 205, was identified and a plasmid, pCN5, was constructed which contains the CpG motif of ODN 205. When administered into turbot via intraperitoneal (i.p.) injection, both ODN 205 and pCN5 could (i) inhibit bacterial dissemination in blood in dose and time dependent manners, and (ii) protect against lethal bacterial challenge. Immunological analyses showed that in vitro treatment with ODN 205 stimulated peripheral blood leukocyte proliferation, while i.p. injection with ODN 205 enhanced the respiratory burst activity, chemiluminescence response, and acid phosphatase activity of turbot head kidney macrophages. pCN5 treatment-induced immune responses similar to those induced by ODN 205 treatment except that pCN5 could also enhance serum bactericidal activity in a calcium-independent manner. To examine whether ODN 205 and pCN5 had any effect on specific immunity, ODN 205 and pCN5 were co-administered into turbot with a Vibrio harveyi subunit vaccine, DegQ. The results showed that pCN5, but not ODN 205, significantly increased the immunoprotective efficacy of DegQ and enhanced the production of specific serum antibodies in the vaccinated fish. Further analysis indicated that vaccination with DegQ in the presence of pCN5 upregulated the expression of the genes encoding MHC class II alpha, IgM, Mx, and IL-8 receptor. Taken together, these results demonstrate that ODN 205 and pCN5 can stimulate the immune system of turbot and induce protection against bacterial challenge. In addition, pCN5 also possesses adjuvant property and can potentiate vaccine-induced specific immunity. (C) 2010 Elsevier Ltd. All rights reserved.

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Twenty microsatellite markers (Po1, Po13, Po33, Po35, Po42, Po48, Po56, Po89, Po91, kop6, kop7, kop8, kop12, kop15, kop18, kop21, kop22, kop23, kop26, Po-strl) were used to assess the meiogynogenetic and mitogynogenetic stocks of the left-eyed flounder, Paralichthys olivaceus, which were derived from single pair crossing. Twelve of the 20 loci utilized showed heterozygosity in the female and were mapped in relation to their centromeres in the meiogynogenetic diploid flounder. Microsatellite-centromere map distance, calculated under the assumption of complete interference, ranged from 15.8 cM for kop22 to 50 cM for Po13, Po56 and Po89. Excluding the kop22, the heterozygosities of the rest of the loci were close to 100%, suggesting the occurrence of near complete interference on the chromosome arms that carried these loci. In the mitogynogenetic diploid flounder, each individual showed exact homozygousity and the segregation profiles did not deviate from the Mendelian 1: 1 pattern. The results indicated that there was no lethal gene linked with the loci analyzed. Such high interference accounted for the high recombination rates and large map distances. The Po13 and Po56 loci, Po91 and kop18 loci, kop15 and kop21 loci are tightly linked on the same chromosome arm in pairs.

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Planktonic foraminiferal delta O-18 record for core DGKS9603 from the Okinawa Trough shows a series of climatic fluctuations and sudden cooling events in short time scale during 50 kaBP, which appear to correlate closely to the Younger Dryas and Heinrich events H1-5 recorded in Chinese loess, the South China Sea, the North Atlantic cores and the Greenland ice cores. Three polarity reversal events, correlating to Gothenburg, Mungo and Laschamp events, approximately correspond to Heinrich events H1, H3 and H5 respectively, which could be a cause of global climate changes. The delta O-18 curve of the Okinawa Trough is well associated with the grain size record of the Lijiayuan loess profile in northwestern China and is somewhat different from the climate fluctuations documented in the Greenland ice cores. These correlation results indicate that regional factors play an important role in controlling the climate changes in the East Asia, and the East Asian Monsoon could be the prominent regional controlling factor.

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海带根是一种治疗糖尿病的民间中药,在沿海地区有很长的民间用药历史。食用海带根能够有效降低糖尿病患者的血糖,起到治疗作用。本文目的在于发现海带根中抗糖尿病的天然活性物质并分析它们在糖尿病治疗中的靶点;进一步开发一种低价且无毒副作用的化学类新药或中药新药。 α-glucosidase和 PTP-1B是II型糖尿病的两个重要靶点,海带根提取物能同时作用于这两个靶点。通过抑制这两种酶,降低血糖水平,85%乙醇粗提物对两种酶的IC50分别为1589ug/ml、IC50 1271ug/ml。乙酸乙酯相和石油醚相分别抑制α-glucosidase和 PTP-1B,IC50分别为380ug/ml和220ug/ml。因此以α-glucosidase和 PTP-1B的抑制活性为导向,用天然产物化学的方法对活性成分进行追踪分离,寻找单体活性物质进而鉴定其结构。由于乙酸乙酯相具有α-glucosidase抑制活性,用硅胶柱层析(石油醚:丙酮5:1、1:1),(二氯甲烷:甲醇60:1、20:1、5:1),凝胶柱层析Sephadex LH20(二氯甲烷:甲醇1:1),HPLC (80% 甲醇-水),对α-glucosidase抑制剂进行分离,得到组分IC50 为3.6ug/ml。用质谱仪和核磁共振确定结构。 生物活性测定结果表明α-glucosidase和 PTP-1B是两种不同的物质,分别位于乙酸乙酯相和石油醚相。光照实验和高温实验表明抑制α-glucosidase的活性成分对光照和温度敏感。光照48h或者50℃ 12h而且对α-glucosidase的抑制活性显著降低,TLC检测并用FeCl3显色初步表明抑制α-glucosidase的活性成分可能是多数酚类物质。动物实验显示在1450ug/kg剂量下,乙酸乙酯相能够显著降低糖尿病小鼠血糖,与阴性对照组差异极显著(P<0.01)。表明,海带根提取物在体内和体外均呈现出抗糖尿病活性,是一种潜在的抗糖尿病药物。

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琼胶是一种从石花菜等红藻中提取的,目前生产工艺和结构等方面研究比较成熟的海藻多糖,广泛应用于医药、仪器等行业。但是,海藻多糖因为具有分子量大,粘度大,溶解度较小的等特点,而使其应用范围受到限制。利用降解的手段对其进行修饰,降低分子量和粘度,改善溶解性,可以拓展其应用范围。并且根据文献报道,琼 胶寡糖具有一些特殊的生物活性,如抗氧化性,抗炎症等。因此,对琼胶降解的研究具有生要意义。本研究中,为了选择一种合适的降解方法,进行了几种水解方法的尝试,其中包括在不同湿度和酸度下盐酸水解,过氧化氢和醋酸催化水解,Fenton体系羟基自由基降解。对于酸水解和Fenton体系氧化还原降解方法,通过粘度法对反应的速度进行了比较,表明氧化还原降解反应中琼胶的粘度降低比较快,并且具有代表性和新意,确定为本实验的降解琼胶的方法并对氧化还原降解所得的产物进行了活性实验。通过模仿自然界普遍存在的氧化还原降解反应,利用Vc诱导的Fenton体系产生的羟基自由基氧化还原降解琼胶得到低分子量的琼胶。降解产物经过高速离心、60%乙醇沉淀,除去分子量比较大的降解产物和磷酸盐,得到可溶于60%乙醇的分子量估计小于3000的降争产物,其产率为85%。利用经Sephadex-G25凝胶色谱分离所香的不同分子量的级分进行分子量和α-葡萄糖苷酶抑制活性关系的实验。降解产物对α-葡萄糖苷酶的抑制率和各级分的浓度呈线性正相关,并且各级分的IC_(50)则随着分子量的降低而降低。另外,对所得的降解产物混合物进行了红外吸收光谱、质子去偶核磁共震碳谱和负离子基质辅助激光诱导-飞行时间质谱结构分析。结果表明,氧化还原降解反应的专一性差,在得到寡糖的同时,在光谱图中出现一些比较复杂的副产物的结构信息。最后,根据MTT法的原理,以有体皮肤成纤维细胞为材料,通过紫外线辐射产生自由基造成氧化损伤,研究降解产物对成纤维细胞的保护作用。当无紫外线辐射时,降解产物对成纤维细胞具有显著的促进生长增殖作用:当经UVa、UBb辐射时则可以显著地表现出对损伤的保护作用,并且这种促进生长和保护作用呈显著的量效关系,表明降解产物具有清除基自由基的作用。但是,因为氧化还原降解以应的机理尚不十分明的以及琼羟胶的特殊结构,使得反应的副产物很难预测,也就使得分离工作难以进行,所以,根据目前所得的信息,尚不能确定是降解产物的什么级分产生的以上两种生物活性。

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血管内皮生长因子(vascular endothelial growth factor, VEGF)是一种多功能的细胞因子,其主要作用是促进血管内皮细胞增殖和增加血管通透性,是肿瘤及正常组织血管生成的中心调控因素,以VEGF为靶点的肿瘤血管靶向性治疗成为近几年肿瘤治疗的新途径。RNAi是近年来新发展的一项反向遗传学技术,是一种研究基因功能的有力工具。斑马鱼作为一种重要的模式生物,被广泛用于胚胎的分子发育机制、疾病模型的构建以及药物筛选等研究中。然而在斑马鱼中运用RNAi技术进行基因功能研究是一个相对较新的领域,研究资料较少,并且目前进行的斑马鱼RNAi实验中,siRNA大都是通过化学方法或体外转录合成的。体外合成的siRNA在进入体内后会被降解而无法达到持久阻抑基因表达的目的。因此本研究旨在探讨VEGF特异性siRNA表达载体对斑马鱼VEGF基因的沉默作用,通过分析表型及相关细胞因子的变化,阐明VEGF对斑马鱼胚胎血管生成的影响及作用机制。 研究通过计算机辅助设计软件,针对斑马鱼VEGF mRNA不同位点设计合成了4段含siRNA特异序列的DNA单链,经退火,克隆入pSilencer 4.1-CMV neo载体CMV启动子下游,构建了重组质粒pS1-VEGF、pS2-VEGF、pS3-VEGF及pS4-VEGF。 通过显微注射的方法将载体导入1-2细胞期斑马鱼体内,于胚胎发育的48 h采用RT-PCR的方法检测VEGF基因的表达量,研究不同干扰序列对VEGF基因表达的干涉作用。结果显示,针对不同位点的表达载体对VEGF基因表达的抑制效率有显著差异。它们对VEGF mRNA的抑制率分别为80.5%,42.8%,12.5%,40.7%。通过筛选我们得到了一条具有高效抑制作用的载体pS1-VEGF,该载体的相应序列靶向斑马鱼两个主要异构体VEGF165和VEGF121的共有外显子序列。 形态学检测结果显示,注射了pS1-VEGF的胚胎出现了心包膜水肿、血流速度减慢、循环红细胞堆积等症状。定量碱性磷酸酶染色显示,注射pS1-VEGF能够抑制斑马鱼胚胎新生血管的形成,当注射剂量为0.4 ng时,血管生成的抑制率为31.8%。NBT/BCIP血管染色显示,注射该载体后72 h,50%的斑马鱼肠下静脉、节间血管以及其它血管的发育受到不同程度的抑制。随着注射剂量的加大,血管发育受抑制的情况也随之加重,当注射剂量为1 ng时,只有心脏、头部及卵黄有血液循环。对干扰效果的特异性进行了研究,结果表明pS1-VEGF对斑马鱼内源基因胸苷酸合成酶(thymidylate synthase, TS)基因的表达没有明显的抑制作用。针对TS基因的shRNA表达载体及与斑马鱼没有同源性的对照载体对VEGF基因表达也没有明显的抑制作用。浓度梯度实验表明在0-1.2 ng的范围内干扰效果具有剂量依赖性。 以胚胎整体原位杂交的方法检测质粒对VEGF基因受体NRP1基因表达的影响,发现VEGF特异性shRNA表达载体能够引起NRP1基因表达的降低,说明斑马鱼中VEGF所介导的血管生成作用至少在部分上是依赖于NRP通路所调节的。 本研究工作为进一步研究斑马鱼基因功能、VEGF调控网络提供了一个快速、有效的手段,为阐明斑马鱼的血管生成机制提供了新的资料,为采用RNAi技术,以VEGF为靶点,以斑马鱼为模型对肿瘤进行基因治疗研究奠定了基础。

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血管生成是肿瘤生长、发展的必经之路,并且与实体瘤的发生、转移有着密切的关系,抑制肿瘤新生血管生成具有特异性高、疗效好、不易产生耐药性以及毒副作用低等特点,因此抑制肿瘤血管形成可望成为治疗癌症的一个突破点,以抗血管生成为主的肿瘤生物治疗研究已成为近十年的研究热点。玻璃海鞘(Coina intestinalis)属于内性目、玻璃海鞘科,因其进化上的独特地位常作为研究神经发育、免疫系统进化的材料。在其它种属的海鞘中分离发现了多种具有抗肿瘤活性的多肽,但关于玻璃海鞘抗血管生成活性多肽的分离纯化和活性研究未见报道。本文利用多种分离纯化手段,采用活性追踪的方法首次从玻璃海鞘中分离得到具有抗新生血管生成作用的多肽,并对其抗血管生成活性做了初步研究。 本研究利用冷丙酮分级沉淀,超滤截留分子量小于5kDa的蛋白,再经SephadexG25、Superdex75柱层析,µRPC C2/C18反相柱层析等分离手段,采用活性追踪的方法,由玻璃海鞘(Coina intestinalis)中分离纯化出抗血管生成多肽PCI,据保留时间计算其分子量为1.8 kDa。MTT检测表明其对人脐静脉血管内皮细胞(HUVEC)具有强烈的抑制作用,IC50为7.5 μg/ml,并对多种肿瘤细胞有直接的抑制作用。斑马鱼胚胎体内实验进一步表明PCI在40 μg/ml的浓度下作用12h,斑马鱼胚胎新生血管生成受到显著抑制,肠下静脉血管长度为正常组的30%,斑马鱼胚胎血管生成率为正常组的45%。

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The toxicity of seven major HAB (harmful algal bloom) species/strains, Prorocentrum donghaiense, Phaeocystis globosa, Prorocentrum micans, Alexandrium tamarense (AT-6, non-PSP producer), Alexandrium lusitanicum, Alexandrum tamarense (ATHK) and Heterosigma akashiwo were studied against rotifer Brachionus plicatilis under laboratory conditions. The results show that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6), or A. lusitanicum could maintain the individual survival and reproduction, as well as the population increase of the rotifer, but the individual reproduction would decrease when exposed to these five algae at higher densities for nine days; H. akashiwo could decrease the individual survival and reproduction, as well as population increase of the rotifer, which is similar to that of the starvation group, indicating that starvation might be its one lethal factor except for the algal toxins; A. tamarense (ATHK) has strong lethal effect on the rotifer with 48h LC50 at 800 cells/mL. The experiment on ingestion ability indicated by gut pigment change shows that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6) and A. lusitanicum can be taken by the rotifers as food, but A. tamarense (ATHK) or H. akashiwo can be ingested by the rotifers. The results indicate that all the indexes of individual survival and reproduction, population increase, gut pigment change of the rotifers are good and convenient to be used to reflect the toxicities of HAB species. Therefore, rotifer is suggested as one of the toxicity testing organisms in detecting the toxicity of harmful algae.

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To investigate harmful effects of the dinoflagellate Alexandrium species on microzooplankton, the rotifer Brachionus plicatilis was chosen as an assay species, and tested with 10 strains of Alexandrium including one known non-PSP-producer (Alexandrium tamarense, AT-6). HPLC analysis confirmed the PSP-content of the various strains: Alexandrium lusitanicum, Alexandrium minutum and Alexandrium tamarense (ATHK, AT5-1, AT5-3, ATC102, ATC103) used in the experiment were PSP-producers. No PSP toxins were detected in the strains Alexandrium sp1, Alexandrium sp2. Exposing rotifer populations to the densities of 2000 cells ml(-1) of each of these 10 Alexandrium strains revealed that the (non-PSP) A. tarnarense (AT-6) and two other PSP-producing algae: A. lusitanicum, A. minutum, did not appear to adversely impact rotifer populations. Rotifers exposed to these three strains were able to maintain their population numbers, and in some cases, increase them. Although some increases in rotifer population growth following exposures to these three algal species were noted, the rate was less than for the non-exposed control rotifer groups. In contrast, the remaining seven algal strains (A. tamarense ATHK, AT5-1, AT5-3, ATC102, ATC103; also Alexandrium sp1 and Alexandrium sp2) all have adverse effects on the rotifers. Dosing rotifers with respective algal cell densities of 2000 cells ml-1 each, for Alexandrium spl, Alexandrium sp2, and A. tamarense strains ATHK and ATC103 showed mean lethal time (LT50) on rotifer populations of 21, 28, 29, and 36h, respectively. The remaining three species (A. tamarense strains AT5-1, AT5-3, ATC102) caused respective mean rotifer LT50S of 56, 56, and 71 h, compared to 160 h for the unexposed "starved control" rotifers. Experiments to determine ingestion rates for the rotifers, based on changes in their Chlorophyll a content, showed that the rotifers could feed on A. lusitanicum, A. minutum and A. tamarense strain AT-6, but could graze to little or no extent upon algal cells of the other seven strains. The effects on rotifers exposed to different cell densities, fractions, and growth phases of A. tamarense algal culture were respectively compared. It was found that only the whole algal cells had lethal effects, with strongest impact being shown by the early exponential growth phase of A. tamarense. The results indicate that some toxic mechanism(s), other than PSP and present in whole algal cells, might be responsible for the adverse effects on the exposed rotifers. (C) 2004 Elsevier B.V. All rights reserved.

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Protein tyrosine phosphatase 1B (PTP1B) plays an important role as a negative regulator in insulin signaling pathways. PTP1B is an effective target for the treatment of type 2 diabetes mellitus. Four bromophenol derivatives from red algae Rhodomela confervoides, 2,2',3,3'-tetrabromo-4,4',5,5'-tetra-hydroxydiphenyl methane (1), 3-bormo-4,5-bis(2,3-dibromo-4,5-dihydroxybenzyl) pyrocatechol (2), bis(2,3-dibromo-4,5-dihydroxybenzyl) ether (3) and 2,2',3-tribromo-3',4,4',5-tetrahydroxy-6'-ethyloxy-methyldiphenylmethane (4) showed significant inhibitory activity against PTP1B (IC50 were 2.4, 1.7, 1.5 and 0.84 mu mol/L, respectively) as potential therapeutical agents for the treatment of type 2 diabetes mellitus. The anti-hyperglycemic effects of the ethanol extracts from R. confervoides on streptozotocin-induced diabetes (STZ-diabetes) in male Wistar rats fed with high fat diet were investigated. The STZ-diabetic rats treated with medium-dose and high-dose alga extracts showed remarkable reductions in fasting blood glucose (FBG) as compared with the STZ-diabetic control. The results indicate that the in vivo anti-hyperglycemic activity of the R. confervoides extracts can be partially attributed to the inhibitory actions against PTP1B of the bromophenol derivatives and that may be of clinical importance in improving the management of type 2 diabetes mellitus.