Amphiphilic sodium alginate-vinyl acetate microparticles for drug delivery

To overcome the fast or burst release of hydrophilic drugs from hydrophilic alginate-based carriers,hydrophobic molecule(vinyl acetate,VAc)was grafted on alginate(Alg),which was further used to prepare drug carriers.Amphiphilic Alg-g-PVAc hydrogel beads were firstly prepared by emulsification/internal gelation technique for the loading of bovine serum albumin(BSA).Then,chitosan was coated on the surface of beads to form novel amphiphilic Alg-g-PVAc/chitosan(Alg-g-PVAc/CS)microcapsules.The BSA-loading amphiphilic Alg-g-PVAc/chitosan(Alg-g-PVAc/CS)microcapsules display similar morphology and size to the hydrophilic alginate/chitosan(AC)microcapsules.However,the drug loading and loading efficiency of BSA in Alg-g-PVAc/CS microcapsules are higher,and the release rate of BSA from Alg-g-PVAc/CS microcapsules is slower.The results demonstrate that the introduction of hydrophobic PVAc on alginate can effectively help retard the release of BSA,and the higher degree of substitution is,the slower the release rate is.In addition,the complex membrane can also be adjusted to delay the release of BSA.As a whole,amphiphilic sodium alginate-vinyl acetate/CS microparticles could be developed for macromolecular drug delivery.     

日本刺参的抗肿瘤及免疫调节作用研究

采用日本刺参（Apostichopus japonicus）浆灌胃移植性肿瘤模型小鼠,探讨了日本刺参的抗肿瘤作用及其对免疫功能的调节作用。结果表明,日本刺参各剂量组均能显著抑制小鼠S180肉瘤的生长,提高小鼠脾指数及胸腺指数（P〈0．05）;显著提高荷瘤小鼠血清溶血素含量（P〈0．01）和抗体形成细胞数（P〈0．05）;提高迟发型变态反应水平和脾淋巴细胞转化能力（P〈0．05,P〈0．01）;促进NK细胞活性和巨噬细胞的吞噬能力（P〈0．05。P〈0．01）。提示日本刺参能全面调节机体的特异性和非特异性免疫功能．控制和杀灭肿瘤细胞。     

重组抗菌肽海藻酸钠微囊制备与体外释放特征研究

对重组表达的海洋生物抗菌肽对虾素3-2进行亲和层析纯化,以海藻酸钠为壁材,采用凝聚法制备了重组抗菌肽海藻酸钠微囊,以微囊的形态和包封率为指标优化制备工艺,对制备的微囊进行体外释放特征的初步研究。结果显示,在氯化钙浓度为1.5%,海藻酸钠浓度为2.0%时,制备的微囊为完整的球形,冷冻干燥后的直径约为1.1 mm,包封率为83.87%。微囊在模拟胃液(pH 2.0)中2 h左右释放量趋于稳定,释放量低于14%;微囊在模拟肠液(pH 7.8)中不断释放,5 h时释放量达98%,表明微囊具有良好的肠溶性而可以抵抗胃液的破坏,可以用作重组抗菌肽缓释/控释制剂,为抗菌肽在水产病害防治过程的口服给药提供实验基础。     

Effects of organochlorine compounds on cytochrome P450 aromatase activity in an immortal sea turtle cell line

Many classes of environmental contaminants affect the reproductive function of animals through interactions with the endocrine system. The primary components affected by endocrine active compounds (EACs) are the steroid receptors and the enzymes responsible for steroidogenesis. This study sought to develop an in vitro model for assessing EAC effects in sea turtles by examining their ability to alter cytochrome P450 aromatase (CYP19) activity. Aromatase is the enzyme responsible for the conversion of testosterone to estradiol. This enzyme is critical in the sexual differentiation of reptiles which demonstrate temperature-dependent sex determination. An immortal testis cell line GST-TS from a green sea turtle was grown in culture at 30 degrees C in RPMI 1640 media. The cells were exposed to three known aromatase inducers; dexamethasone (Dex), 8Br-cyclic AMP, or human chronic gonadotropin (HCG) and one aromatase inhibitor 4-androstenol-dione (4-OHA). In addition, the GST-TS cells were exposed to 0.1-30 microM atrazine and 3-100 microM 4,4'-DDE. The inducing compounds that have been shown to increase aromatase activity in other systems failed to induce aromatase activity in the GST-TS cells, yet exposure to the inhibiting compound, 4-OHA, did result in a significant reduction. Atrazine (0.1, 1.0 and 10 microM) significantly induced aromatase activity following a 24 h exposure, and 4,4'-DDE inhibited the activity but only at cytotoxic concentrations (100 microM). Based on these results, this in vitro model can be useful in examining the endocrine effects of EACs in sea turtles.     

营养盐亏缺与恢复对威氏海链藻(Thalassiosira weissflogii)生长和生化组成的影响

通过室内纯培养,研究了营养盐亏缺以及解除亏缺对威氏海链藻 ( Thalassiosira weissflogii ) 的细胞形态、细胞增殖、胞内蛋白质、可溶性糖含量变化的影响,考察了藻细胞生长和生化组成的恢复情况.结果表明,在多种营养盐亏缺 ( N 亏缺、P 亏缺、Si 亏缺、N + P 亏缺、贫营养 ) 的条件下,威氏海链藻生长受到不同程度的抑制,全部受试细胞形态均出现不同程度变异,细胞增殖显著低于正常营养供给.恢复营养,细胞形态的损伤能得以有效恢复,细胞增殖能力得以一定程度的有限恢复,其中氮、磷亏缺的恢复效应最为明显.营养盐亏缺,各处理组细胞生化组成显著低于正常培养状态.恢复营养,氮、磷亏缺组藻细胞生化组成仍显著低于正常营养供给,生化组成的恢复效应不明显,生理活性无法恢复,而硅盐亏缺的藻细胞生长和生化组成均能恢复到正常水平.实验证明,微藻细胞的生化组成比形态变化与增殖更能反映营养盐亏缺,可作为微藻受氮、磷营养盐限制的指标.     

从培养的微藻中初步筛选抗菌和抗肿瘤活性物质

从8种微藻中制备水溶和脂溶性的提取物,用来筛选抗菌和抗肿瘤活性.抗菌活性是用抑菌豳法(细菌用金黄色葡萄球菌,枯草芽孢杆菌和大肠杆菌为指示菌,真菌用黑曲霉和产黄青霉为指示菌).实验选用的所有的微藻均有不同程度的抗一种或多种菌活性,抗金黄色葡萄球菌微藻提取物中的57.14%有抗耐甲氧西林金黄色葡萄球菌活性,采用MTT法体外...     

Cellular responses in the mussel Mytilus edulis following exposure to diesel oil emulsions: Reproductive and nutrient storage cells

The objectives of this study were to examine, and quantify with the aid of stereological techniques,^1,2 the effects of exposure to hydrocarbons on the reproductive and nutrient storage cell systems in the marine mussel, Mytilus edulis. In addition, the capacity for recovery of these cell systems was quantified following a period of hydrocarbon depuration. The second phase of these investigations examined the significance of nutritional reserves at the time of exposure to hydrocarbons and how this may influence the animal's ability to survive the insult. The results (Table 1) indicated that, first, hydrocarbons had a deleterious effect on the nutritional storage cells leading to reduced fecundity and oocyte atresia (degeneration). Given a period of depuration, however, there were indications of a recovery in both the reproductive and storage cell systems. Second, the ability of mussels to survive hydrocarbon exposure was dependent upon the nutrient reserves at the onset of exposure.     

LPS-induced NO inhibition and antioxidant activities of ethanol extracts and their solvent partitioned fractions from four brown seaweeds

The nitric oxide inhibitory (NOI) and antioxidant (ABTS and DPPH radical scavenging effects with reducing power) activities of the ethanol (EtOH) extracts and solvent partitioned fractions from Scytosiphon lomentaria, Chorda filum, Agarum cribrosum, and Desmarestia viridis were investigated, and the correlation between biological activity and total phenolic (TP) and phlorotannin (TPT) content was determined by PCA analysis. The yield of EtOH extracts from four brown seaweeds ranged from 2.6 to 6.6% with the highest yield from D. viridis, and the predominant compounds in their solvent partitioned fractions had medium and/or less polarity. The TP and TPT content of the EtOH extracts were in the ranges of 25.0–44.1 mg GAE/g sample and 0.2–4.6 mg PG/g sample, respectively, which were mostly included in the organic solvent partitioned fractions. Strong NOI activity was observed in the EtOH extracts and their solvent partitioned fractions from D. viridis and C. filum. In addition, the EtOH extract and its solvent partitioned fractions of D. viridis exhibited little cytotoxicity to Raw 264.7 cells. The most potent ABTS and DPPH radical scavenging capacity was shown in the EtOH extracts and their solvent partitioned fractions from S. lomentaria and C. filum, and both also exhibited strong reducing ability. In the PCA analysis the content of TPT had a good correlation with DPPH (r = 0.62), ABTS (r = 0.69) and reducing power (r = 0.65), however, an unfair correlation was observed between the contents of TP and TPT and NOI, suggesting that the phlorotannins might be responsible for the DPPH and ABTS radical scavenging activities.     

微小卡罗藻溶血活性的生长期特征及其溶血毒素种类分析

研究微小卡罗藻的生长曲线及不同生长期藻细胞提取液和去藻上清液溶血活性的大小,通过乙酸乙酯和Bligh-Dyer两种方法提取溶血毒素,利用柱层析和薄层层析对溶血毒素成分进行初步的分离与分析。结果表明,藻细胞提取液在4个生长阶段中都具有明显的溶血活性,去藻上清液在平稳期和衰亡期具有明显的溶血活性,延滞期及对数生长期溶血活性很低,表明微小卡罗藻的溶血物质是其自身正常生理代谢产物,且在细胞老化过程中可以释放至水体环境中。两种提取方法都可以将溶血毒素提取出来,进一步的色谱分离和生物活性检测发现微小卡罗藻主要含有3种不同性质的溶血毒素,其溶血作用可能与微小卡罗藻较高含量的多不饱和脂肪酸18:5n3和22:6n3有关。     

文蛤多肽的体外抗癌活性研究

采用硫酸铵沉淀、有机溶剂分离、Sephadex G-25分子筛层析等实验方法,从文蛤肉中提取了一种低分子量的多肽,命名为Mer2．本文以溴化二苯偶氮盐（MTT）法检测Mer2对体外培养的癌细胞的抑制作用．结果表明Mer2对体外培养的人肝癌细胞株（HepG2）、宫颈癌细胞株（Hela）、胆管癌细胞株（QBC939）、肺癌细胞株（SPC—A-1和LTEP—a-2）的生长均有很强的抑制作用,且抑制效果随着Mer2含量的增高和处理时间的延长而增强,证明Mer2具有广谱的抗癌活性．其中Mer2对人肝癌HepG2细胞株抑制作用最为显著,光学显微镜观察经Mer2细胞培养液处理后的细胞形态发生明显的改变,流式细胞仪实验的结果表明处理后的细胞周期发生明显的改变,并在G0／G1期前出现凋亡峰．     

5-氟尿嘧啶壳聚糖缓释微囊制备工艺研究

5-氟尿嘧啶 (5- Fu)以壳聚糖为载体 ,采用乳化化学交联技术 ,制备了 5- Fu壳聚糖微囊。用正交设计安排试验 ,以微囊载药量、药物包封率、粒子分布百分数为指标优化微囊的制备工艺 ,并以最佳制备工艺条件制得含药微囊 ,方法重现性好 ,制备工艺稳定。体外释放度试验表明 ,该微囊具有较好的缓释作用。     

Field methods for amending marine sediment with activated carbon and assessing treatment effectiveness

Previous laboratory studies have shown reductions in PCB bioavailability for sediments amended with activated carbon (AC). Here we report results on a preliminary pilot-scale study to assess challenges in scaling-up for field deployment and monitoring. The goals of the preliminary pilot-scale study at Hunters Point Shipyard (San Francisco, USA) were to (1) test the capabilities of a large-scale mixing device for incorporating AC into sediment, (2) develop and evaluate our field assessment techniques, and (3) compare reductions in PCB bioavailability found in the laboratory with well-mixed systems to those observed in the field with one-time-mixed systems. In this study we successfully used a large-scale device to mix 500kg of AC into a 34.4m(2) plot to a depth of 1ft, a depth that includes the majority of the biologically active zone. Our results indicate that after 7 months of AC-sediment contact in the field, the 28-day PCB bioaccumulation for the bent-nosed clam, Macoma nasuta, field-deployed to this AC-amended sediment was approximately half of the bioaccumulation resulting from exposure to untreated sediment. Similar PCB bioaccumulation reductions were found in laboratory bioassays conducted on both the bivalve, M. nasuta and the estuarine amphipod, Leptocheirus plumulosus, using sediment collected from the treated and untreated field plots one year after the AC amendment occurred. To further understand the long-term effectiveness of AC as an in situ treatment strategy for PCB-contaminated sediments under field conditions, a 3-year comprehensive study is currently underway at Hunters Point that will compare the effectiveness of two large-scale mixing devices and include both unmixed and mixed-only control plots.     

利用酒精酵母细胞表面展示技术制备活疫苗及其潜在应用

最近几年,出现把有关的基因在宿主表达之后的目的蛋白分泌并固定在细菌、病毒和酵母菌细胞表面上的技术一表面展示技术。在这些宿主中,酒精酵母菌是最佳的微生物。本文对利用酒精酵母细胞表面展示技术制备活疫苗的基本原理,在酵母细胞表面展示蛋白质的优点,以及哈维氏弧菌溶血素蛋白在酵母菌细胞表面上展示及其作为活疫苗的潜在应用作了综述。研究结果表明在酒精酵母菌细胞表面上展示的哈维氏弧菌溶血素蛋白在海洋动物中具有很好的免疫原性和免疫保护性。     

Catalytic function of avian cytochrome P450 1A4 and 1A5 (CYP1A4 and CYP1A5) enzymes heterologously expressed using in vitro yeast system

The present study clarifies the enzymatic properties of two avian cytochrome P4501A (CYP1A) paralogs, CYP1A4 and 1A5, using a yeast-based vector system. Recombinant CYP1A4 and 1A5 proteins from common cormorant (Phalacrocorax carbo) were expressed in yeast cells, and showed typical reduced CO-difference spectra with a peak at 446 nm. Kinetic analysis of O-dealkylase of methoxy-, ethoxy-, pentoxy- and benzyloxyresorufin catalyzed by the CYP1A enzymes revealed that Vmax value for ethoxyresorufin-O-deethylase (EROD) activity was much higher than that for the other three O-dealkylase activities for both isozymes. Interestingly, remarkable substrate specificity of the CYP1As was observed for O-dealkylation of benzyloxyresorufin and methoxyresorufin; CYP1A4 was highly specific for catalyzing benzyloxyresorufin-O-debenzylase activity, whereas CYP1A5 was more efficient in catalyzing methoxyresorufin-O-demethylase activity. The present study also measured CYP1A-dependent EROD activity in the presence of 2,3,7,8-tetrachlorodibenzofuran (TCDF) to evaluate the ability of this dioxin-like congener to inhibit the EROD activity. One hundred nanomolar TCDF noncompetitively inhibited CYP1A5-dependent EROD activity, although no inhibitory effect was detected for CYP1A4-dependent EROD activity. These results indicate that the avian CYP1A paralogs have different affinities for substrate and inhibitor, thus suggesting their distinct physiological and toxicological roles.     

微绿球藻固定化培养及其对对虾养殖水质调控

用褐藻酸钙作固定化载体,初步探讨了胶球直径、不同接种量、CaCl2溶液浓度、褐藻酸钠浓度等固定化条件对微绿球藻(Nannochloris oculata)生长及其对养殖水质的影响。结果表明,微绿球藻固定化培养的最好条件是胶球的直径为2.5mm、微藻接种量为1×106 Cell/mL, CaCl2溶液的质量分数为3%,褐藻酸钠溶液质量分数为2%。在此条件下制备的固定化藻珠, 微绿球藻生长率较高,生长周期较长。实验期间固定化微绿球藻的生物量增加了约17倍,证明了它的生理活性不会因固定化而受干扰。引入固定化的微绿球藻不但可以降低养殖水体中氨氮、亚硝酸氮等有害因子的浓度,还能提高水中溶解氧含量,使水体环境长时间处于良好的动态平衡状态。     

In vitro screening for endocrine disruptive activity in selected South African harbours and river mouths

Various waterborne anthropogenic contaminants disrupt the endocrine systems of wildlife and humans, targeting reproductive pathways, among others. Very little is known, however, regarding the occurrence of endocrine disruptive activity in South African freshwater ecosystems, and coastal ecosystems have not been studied in this regard. In a first attempt to investigate endocrine disruptive activity in South African coastal waters, surface water samples collected from harbours, river mouths and estuaries in three metropolitan municipalities, eThekwini (which includes Durban), Nelson Mandela (specifically Port Elizabeth Harbour) and City of Cape Town, were screened for (anti) oestrogenicity and (anti)androgenicity using recombinant yeast bioassays. Moreover, levels of the female hormone 17β-(o)estradiol (E2) were determined in all samples, as well as a selection of hydrocarbons in the eThekwini samples. A high proportion of samples collected from eThekwini were oestrogenic, whereas none from Port Elizabeth Harbour and only a single river mouth sampled in the City of Cape Town were oestrogenic. E2 was detected in all the samples tested, but at higher concentrations at the eThekwini and City of Cape Town localities than Port Elizabeth Harbour. In addition, the recombinant yeast assays revealed that anti-androgenicity was widespread, being detected in the majority of samples screened apart from those representing Port Elizabeth Harbour. Conversely, no anti-oestrogenic or androgenic activity was detected. Anti-androgenicity did not associate with hydrocarbon loads, providing evidence that other anti-androgens were responsible for the observed activity. The present data suggest potential reproductive disruption in marine and estuarine fauna inhabiting the eThekwini and City of Cape Town regions.     

Microencapsulation of okadaic acid as a tool for studying the accumulation of DSP toxins in mussels

The possibility and effectiveness of microencapsulation of okadaic acid (OA) in gelatin-acacia microcapsules has been studied. The encapsulation efficiency was higher than 33%. The microcapsules were shown to be very stable, not leaching more than 9% of the encapsulated OA in a 20-h period. OA from the microcapsules was absorbed by the mussels very efficiently, accumulating--after 3 days of feeding and one of depuration--65% of the OA in microcapsules and 22% of the total OA used at the beginning of the microencapsulation process. These efficiencies and the possibility of encapsulating single DSP toxins and derivatives constitute a valuable tool for the study of the accumulation and biotransformation of DSP toxins in bivalves.     

不同氮源对微小亚历山大藻生长和毒素产生的影响

通过尿素、氯化铵、酵母浸出粉和硝酸钠等氮源对微小亚历山大藻(Alexandrium minutum)生长及毒素产生的影响研究,分析了微小亚历山大藻对不同氮源利用状况的差异.结果表明,在氮饥饿条件下,加入硝酸钠和酵母浸出粉能显著促进微小亚历山大藻的生长;高浓度的氯化铵在加入后对微小亚历山大藻有一定的毒性效应,表现为生长停滞,但毒性效应在5 d后消失,并得到与添加硝酸钠及酵母浸出粉相似的增长速率0.21 d-1;添加尿素对微小亚历山大藻的生长没有显著促进作用.在四种氮源中,尿素对微小亚历山大藻毒素产生的刺激作用也最弱,在稳定期每个细胞藻细胞毒素含量维持在6.00~8.00 fmol;添加硝酸钠、氯化铵和酵母浸出粉的藻细胞在稳定期毒素含量分别达到11.85,12.86和14.64 fmol.硝酸钠和氯化铵刺激藻毒素产生的效果比酵母浸出粉更为直接.四种含氮营养盐对微小亚历山大藻毒素组成的影响都很小.