Toxic Effects of Benzo[a]pyrene(BaP) on the Parameters and Genes Associated with Innate Immune Defense of Pagrus major

苯并[a]芘(BaP)在海洋环境中广泛存在，是一种重要的环境污染物，具有免疫毒性和致癌性。高等生物的免疫系统对异源性物质十分敏感，能够对外界环境变化作出迅速的反应。先天性免疫系统是鱼类重要的防御体系，环境污染物对免疫系统的毒性作用直接影响到鱼类的健康与种群繁衍，因而研究污染物对鱼类免疫毒性效应具有重要的科学意义。本项研究选择典型的环境污染物BaP为受试物，以海水重要养殖鱼类真鲷作为实验材料，利用分子生物学、免疫学和毒理学等技术，从基因、蛋白等不同水平对比研究了真鲷胚胎、仔鱼及幼鱼不同发育阶段不同免疫学参数随BaP暴露剂量、时间的动力学变化，并重点探讨了鱼类先天性免疫中重要的免疫因子抗菌肽hep...Benzo[a]pyrene (BaP) exists widely and is therefore an important environmental contaminant, with carcinogenic and immunotoxic properties. The immune system in higher organisms is under integrated control and has the capacity to rapidly respond to the environment, which is very sensitive to xenobiotic and regarded as a target for environmental xenobiotics. The innate immune system of fish is the fi...学位：理学博士院系专业：海洋与环境学院环境科学与工程系_环境科学学号：2122005140318

The Effects of Benzo [a] Pyrene (BaP) Exposure on the CYP1A1 mRNA and AhR2 mRNA Expression of Red Seabream(Pagrus major)

通过水体暴露方式对海水养殖真鲷进行bAP持续染毒,利用实时定量PCr技术研究了真鲷细胞色素P450基因(CyP1A1)和芳香烃受体基因(AHr2)随bAP暴露剂量、时间的动力学变化。结果发现,0.1~1.5μg/l环境浓度的bAP能够显著性诱导CyP1A1基因和AHr2基因的表达,且AHr2 MrnA早于CyP1A1 MrnA被诱导表达;bAP持续暴露48 H,CyP1A1和AHr2基因的表达水平均随暴露时间的延长而显著升高,染毒72 H后又回复到本底水平,实验表明这两个基因的表达与bAP的暴露剂量和暴露时间之间具有显著性的剂量-效应和时间-效应关系。The gene expression patterns of CYP1A1 and Aryl hydrocarbon receptor(AhR2) of red seabream(Pagrus major) were both measured using real-time quantitative PCR(qPCR) when fish exposed to environmentally relevant concentration of BaP(0.1,0.5 and 1.0 μg/L,respectively).The results showed that CYP1A1 mRNA and AhR2 mRNA could be induced significantly,besides,the time of AhR2 mRNA induced ahead of the time of CYP1A mRNA induced.The two genes were induced markedly at the begining of BaP exposure,and then decreased to the basal levels after 72 h.The results demonstrate that BaP can regulate CYP1A1 and AhR2 transcript in a dose and time dependent manner.国家“863”计划重点资助项目(2007AA091406);国家自然科学基金资助项目(30770391

A Study on the Cytotoxic and Genotoxic Effects of Rare Earth Multi-element Compound Fertilizer (RE-MECF) on Zea mays L.

为了研究常乐稀土多元复合肥对主要农作物--玉米(Zea mays L.)是否产生细胞毒性和遗传毒性效应,采用不同梯度质量浓度的稀土复合肥溶液对玉米进行处理,观察其根尖生长状况,统计分析细胞死亡率、微核率、有丝分裂指数以及对细胞DNA的损伤情况.结果表明,质量浓度为10mg·L-1以下时,常乐稀土复合肥能够促进玉米根尖的生长,100mg·L-1以上时抑制根尖生长;随着试验浓度的递增,细胞死亡率、微核率、DNA损伤率逐渐上升并具有正相关性,表现出明显的剂量-效应关系,而有丝分裂指数却随着浓度的增加逐渐下降.以上结果表明,常乐稀土复合肥对玉米确有一定的细胞毒性和遗传毒性效应,其细胞毒性阈值和遗传毒性阈值皆为100mg·L-1.The root tips were treated with different concentrations of Rare Earth Multi-element Compound Fertilizer(RE-MECF)in order to evaluate the effect on maize in terms of the growth of root-tip, the mortality, the frequency of micronucleus(FMN),mitosis index(MI)and DNA damage. The results indicated that the dosage below 10mg·L-1 (expressed by concentration of RE-MECF) could accelerate the growth of root tips of Zea mays L., but the groups which were treated by the dosages above 100mg·L-1 were repressed. The mortality, frequency of micronucleus(FMN)and DNA damage increased while mitosis index(MI)decreased with increasing concentrations, from which, the dosage-effect relationship was clearly demonstrated. These results suggested that the RE-MECF had certain cytotoxic and genotoxic effect, and that the cytotoxic and genotoxic threshold dosages were both 100mg·L-1.国家自然科学基金项目(No.30470421);; 江苏省高等学校自然科学基金项目(No.D180215

A male-specific expression gene, encodes a novel anionic antimicrobial peptide, scygonadin, in Scylla serrata

【英文摘要】Scygonadin is a novel antimicrobial peptide, which was originally isolated from the seminal plasma of the mud crab, Scylla serrata. Based on the partial 20-residue NH2-terminal sequence of the peptide, H-Gly-Gln-Ala-Leu-Asn-Lys-Leu-Met-Pro-Lys-Ile-Val-Ser-Ala-Ile-Ile-Tyr-Met-Val-Gly-OH, scygonadin was cloned from the gonads of S. serrata using a degenerated reverse transcriptase (RT)-PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA sequence contains an open reading frame of 539 bases (excluding polyA) with a coding capacity of 126 amino acids, which constitutes a putative NH2-terminal signal sequence (1–24) and a mature peptide (25–126). Analysis of the genomic DNA sequence revealed that scygonadin consists of 2300 bp containing two introns (1569 and 120 bp) and three exons (187, 131 and 218 bp) and this sequence is different from any other reported antimicrobial peptide. The theoretical pI of the mature peptide is 6.09, which suggests that it is an anionic molecule. The sex and tissue-specific expression of the scygonadin gene was revealed using RT-PCR and Northern-blot analysis of multiple tissues of S. serrata males and females and this demonstrated that the scygonadin gene was predominantly expressed in the male reproductive tract of S. serrata and was restricted to the ejaculatory duct. This suggests that scygonadin might be one of the antibacterial peptides responsible for protection of the male crab reproductive tract from invading pathogenic microorganisms, so as to maintain a sterile environment leading to successful fertilization

硅双极晶体管的低温h_（FE）

低温下h_(FE)主要决定于发射效率γ和集电区倍增因子M。影响γ的是禁带收缩和基区费米能级。M则主要由中性杂质被碰撞电离而造成的电流倍增效应决定，并且这是一种自抑制效应。以上观点与实验结果基本相符合

Comparative study of 17 β-estradiol on endocrine disruption and biotransformation in fingerlings and juveniles of Japanese sea bass Lateolabrax japonicus

Estrogenic contaminants in the aquatic environment are associated with endocrine disruption and feminization in fish. The effects of endocrine disrupting chemicals (EDCs) on fish have been well documented. However, very few studies have focused on 17 β-estradiol (E2) and its effects on endocrine system and biotransformation in a single prolonged exposure. This study investigated changes in the levels of serotonin (5-hydroxytryptamine) and acetyl choline esterase (AchE) in brain, cortisol in plasma and Ethoxyresorufin-O-deethylase (EROD) activity in gill of two different size groups (fingerlings and juveniles) of Japanese sea bass (Lateolabrax japonicus) upon exposure to two sub-lethal concentrations (200 and 2000 ng L-1) of E2 for 30d. The results indicate that cortisol level and EROD activity significantly increased in both groups, whereas serotonin level increased in juveniles and decreased in fingerlings due to E2 exposure. The correlation analysis revealed that E2 significantly affected the endocrine and biotransformation systems in both age groups. ? 2014 Elsevier Ltd

Regulation of metallothionein gene expression in response to benzo[a]pyrene exposure and bacterial challenge in marine cultured black porgy (Acanthopagrus schlegelii)

Metallothioneins (MTs) are normally considered to be sensitive indicators of heavy metal pollution, but it is less clear whether the MT gene can actively respond to other environmental stresses. In this study, an MT mRNA molecular sequence of 471 bp (full length) was identified in marine cultured black porgy (Acanthopagrus schlegelii), encoding 60 amino acids containing 20 cysteine residues. The MT sequence was highly homologous to that of other fish belonging to the MT superfamily type 1 family. The three dimensional structure of the deduced MT peptide was composed of two metal-binding domains capable of ligating divalent heavy metals. The MT mRNA transcripts were detected in the 11 tested tissues and the highest quantity was present in the liver. Stresses by two factors, benzo[a]pyrene (B[a]p) exposure and bacterial challenge, were evaluated on MT gene expression. The level of MT gene transcripts in the liver significantly declined 24 h post B[a]p exposure and the quantity was significantly correlated with the exposure time during a 24 h period. In contrast, MT gene expression in the liver was significantly increased 48 h post bacterial infection and the quantity was significantly correlated with the infection time during this period of 48 h. Our results indicated that MT gene expression in black porgy liver was sensitive to environmental stresses other than just the heavy metal pollution reported, suggesting that the development of a reliable biomarker for heavy metal pollution will be more complex than expected

Modulation of immune-associated parameters and antioxidant responses in the crab (Scylla serrata) exposed to mercury

Organic and inorganic contaminants can suppress immune function in molluscs and crustaceans. It was postulated that metals could modulate immune function in marine crabs. To test this hypothesis, sublethal effects of mercury (Hg) on cellular immune and biochemical responses of crabs were determined. When crabs were exposed for 14. d to environmentally-relevant concentrations of Hg, changes in immune-associated parameters including, total haemocyte count, lysosomal membrane stability, phenoloxidase, super oxide generation and phagocytosis were observed. Oxidative stress, as measured by lipid peroxidation, antioxidant responses, including superoxide dismutase and catalase activities and glutathione-mediated antioxidant enzymes in serum, haemocyte lysate, gills, hepatopancreas and muscle were assessed in crabs exposed to Hg. Exposure to Hg resulted in significantly lesser immune-associated parameters in haemolymph and antioxidants in all tissues studied. Conversely, GST and phenoloxidase activity, were greater in crabs exposed to Hg. Responses of antioxidant parameters (SOD, CAT and GP. x) were positively correlated with immune responses, including THC, superoxide and phagocytosis. These results were postulated to be due to an immediate response of antioxidant defense to oxygen radicals generated. Overall, the results suggest that 14. d exposure to environmentally realistic concentrations of Hg causes immunomodulation and potentially harmful lessened antioxidant defenses of crabs. ? 2012 Elsevier Ltd

动态惩罚函数非线性规划遗传算法及在汽车变速器中的应用

针对传统遗传算法在求解非线性规划问题时局部搜索能力较弱,惩罚函数求解精度不高的缺陷,将非线性规划算法引入到遗传算法中,提出一种基于动态惩罚函数的非线性规划遗传算法,将遗传算法的全局寻优能力和非线性规划算法的局部寻优能力结合起来,并引入动态惩罚函数,根据不可行点到可行域的距离和可行度自适应的调整惩罚项的值,从而能够快速求出全局最优解。介绍了动态惩罚函数的设计、改进遗传算法的关键技术和流程。最后,以某型号汽车变速器的优化设计验证了算法的合理性。与传统遗传算法相比,改进后的遗传算法解的质量、收敛速度明显提高,因而为遗传算法的改进提供了一种新的思路