Plasma biochemical responses of the planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) to prolonged toxic cyanobacterial blooms in natural waters

The planktivorous filter-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) are the attractive candidates for bio-control of plankton communities to eliminate odorous populations of cyanobacteria. However, few studies focused on the health of such fishes in natural water body with vigorous toxic blooms. Blood parameters are useful and sensitive for diagnosis of diseases and monitoring of the physiological status of fish exposed to toxicants. To evaluate the impact of toxic cyanobacterial blooms on the planktivorous fish, 12 serum chemistry variables were investigated in silver carp and bighead carp for 9 months, in a large net cage in Meiliang Bay, a hypereutrophic region of Lake Taihu. The results confirmed adverse effects of cyanobacterial blooms on two phytoplanktivorous fish, which mainly characterized with potential toxicogenomic effects and metabolism disorders in liver, and kidney dysfunction. In addition, cholestasis was intensively implied by distinct elevation of all four related biomarkers (ALP, GGT, DBIL, TBIL) in bighead carp. The combination of LDH, AST activities and DBIL, URIC contents for silver carp, and the combination of ALT. ALP activities and TBIL, DBIL. URIC concentrations for bighead carps were found to most strongly indicate toxic effects from cyanobacterial blooms in such fishes by a multivariate discriminant analysis. (C) 2009 Elsevier B.V. All rights reserved.

EFFECTS OF IRON ON PHYSIOLOGICAL AND BIOCHEMICAL CHARACTERISTICS OF Microcystis wesenbergii (Kom.) Kom. (Cyanobacterium)

Iron is an essential trace element for biological requirements of phytoplankton. Effects of iron on physiological and biochemical characteristics of Microcystis wesenbergii were conducted in this study. Results showed that 0.01 mu M [Fe3+] seriously inhibited growth and chlorophyll synthesis of M. wesenbergii, and induced temporary increase of ATPase activities, however, NR. ACP and ALP activities were restrained by iron limitation. Interestingly, iron addition on day 8 resulted in the gradual restoration of structures and functions of above enzymes and resisted a variety of stresses from iron limitation. M. wesenbergii in 10 mu M [Fe3+] treatment group grew normally. enzymes maintained normal levels, and residual phosphate contents in cultures first sharply decreased, then smoothly as M. wesenbergii has a characteristic of luxury consumption of phosphorus. Above parameters in 100 mu M [Fe3+] treatment group were almost same with those in 10 mu M [Fe3+] treatment group except for NR, ACP and ALP activities. In 100 mu M [Fe3+] treatment group, activities of ACP and ALP had temporary increase because phosphate and ferric iron could form insoluble compound - ferric phosphate (Fe3PO4) through adsorption effect. resulting in lack of bioavailable phosphate in culture media. The experiment suggested that too low or too high iron can affect obviously physiological and biochemical characteristics of M. wesenbergii.

Plasma biochemical responses of the omnivorous crucian carp (Carassius auratus) to crude cyanobacterial extracts

Healthy crucian carp (Carassius auratus) were treated by intraperitoneal (i.p.) injection of crude cyanobacterial extracts at two doses, 50 and 200 mu g MC-LR equiv kg(-1) BW. High mortality (100%) was observed within 60 h post injection in the high-dose group. In the treated fish, activities of four plasma enzymes, alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH), all showed substantial increases, with both dose and time-dependent effects. These increases of enzyme activity indicate severe impairment occurred in the liver of crucian carp over time. Plasma concentrations of energy-related biomolecules including glucose (GLU), cholesterol (CHO), triglyceride (TG), and total protein (TP) showed marked changes in the high-dose group, possibly a nutritional imbalance correlated with the liver injury caused by intraperitoneal exposure to crude cyanobacterial extracts.

Dietary phosphorus requirement of juvenile black seabream, Sparus macrocephalus

A growth trial was conducted to estimate the optimum requirement of dietary available phosphorus (P) for black seabream (Sparus macrocephalus) in indoor net cages (1.5x1.0x1.0 m). Triplicate groups of black seabream (11.45 +/- 0.02 g) were fed diets containing graded levels (0.18, 0.36, 0.54, 0.72, 0.89 and 1.07%) of available P to satiation for 8 weeks. The basal diet (diet 1), containing 0.18% available P, was supplemented with graded levels of monosodium phosphate (NaH2PO4 2H(2)O) to formulate five experimental diets. The fish were fed twice daily (08:00 h and 16:00 h) and reared in seawater (salinity, 26-29 g l(-1)) at a temperature of 28 +/- 1 degrees C. Dissolved oxygen during the experiment was above 5 mg l(-1). The specific growth rate (SGR), weight gain (WG), feed efficiency (FE) and protein efficiency ratio (PER) were all significantly improved by dietary phosphorus up to 0.54% (P<0.05) and then leveled off beyond this level. Hepatosomatic index (HSI) was inversely correlated with dietary phosphorus levels (P< 0.05). Efficiency of P utilization stabled in fish fed diets containing 0.18%-0.54% available P and then decreased dramatically with further supplementation of dietary phosphorus. Body composition analysis showed that the whole-body lipid, ash, calcium and phosphorus contents were all significantly affected by dietary available P concentration (P<0.05), however, no significance were found in whole-body calcium/phosphorus (Ca/P) ratios among all the treatments (P>0.05). Dietary phosphorus levels also affected the mineralization of vertebrae, skin and scale (P<0.05). Ca/P ratios in vertebrae and scale were not influenced by dietary P supplementation, while skin Ca/P ratio increased statistically with dietary available P levels (quadratic effect, P<0.001). The blood chemistry analysis showed that dietary available P had distinct effects on enzyme activities of alkaline phosphatase (ALP) and plasma lysozyme (LSZ), as well as contents of triacyglycerol (TG) and total cholesterol (T-CHO) (P<0.05). Broken-line analysis showed maximum weight gain (WG) was obtained at dietary available P concentrations of 0.55%. Quadratic analysis based on P contents in whole fish, vertebrae or scale indicated that the requirements were 0.81, 0.87 and 0.88%, respectively. Signs of phosphorus deficiency were characterized by poor growth, slightly reduced mineralization and an increase in body lipid content. (C) 2008 Published by Elsevier B.V.

Primary cultured cells as sensitive in vitro model for assessment of toxicants-comparison to hepatocytes and gill epithelia

In an effort to develop cultured cell models for toxicity screening and environmental biomonitoring, we compared primary cultured gill epithelia and hepatocytes from freshwater tilapia (Oreochromis niloticus) to assess their sensitivity to AhR agonist toxicants. Epithelia were cultured on permeable supports (terephthalate membranes, "filters") and bathed on the apical with waterborne toxicants (pseudo in vivo asymmetrical culture conditions). Hepatocytes were cultured in multi-well plates and exposed to toxicants in culture medium. Cytochrome P4501A (measured as 7-Ethoxyresorufin-O-deethylase, EROD) was selected as a biomarker. For cultured gill epithelia, the integrity of the epithelia remained unchanged on exposure to model toxicants, such as 1,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), benzo(a)pyrene B[a]P, polychlorinated biphenyl (PCB) mixture (Aroclor 1254), and polybrominated diphenyl ether (PBDE) mixture (DE71). A good concentration-dependent response of EROD activity was clearly observed in both cultured gill epithelia and hepatocytes. The time-course response of EROD was measured as early as 3 h, and was maximal after 6 h of exposure to TCDD, B [alp and Aroclor 1254. The estimated 6 h EC50 for TCDD, B [a]P, and Aroclor 1254 was 1.2x10(-9), 5.7x10(-8) and 6.6x10(-6) M. For the cultured hepatocytes, time-course study showed that a significant induction of EROD took place at 18 h, and the maximal induction of EROD was observed at 24 h after exposure. The estimated 24 It EC50 for TCDD, B[a]P, and Aroclor 1254 was 1.4x10(-9), 8.1x10(-8) and 7.3x10(-6) M. There was no induction or inhibition of EROD in DE71 exposure to both gill epithelia and hepatocytes. The results show that cultured gill epithelia more rapidly induce EROD and are slightly more sensitive than cultured hepatocytes, and could be used as a rapid and sensitive tool for screening chemicals and monitoring environmental AhR agonist toxicants. (c) 2006 Elsevier B.V. All rights reserved.

INTERVALLEY-GAMMA-CHI DEFORMATION POTENTIALS IN III-V ZINCBLENDE AND SI SEMICONDUCTORS BY AB-INITIO PSEUDOPOTENTIAL CALCULATIONS

Intervalley GAMMA - X deformation potential constants (IVDP's) have been calculated by first principle pseudopotential method for the III-V zincblende semiconductors AlP, AlAs, AlSb, GaP, GaAs, GaSb, InP, InAs and InSb. As a prototype crystal we have also carried out calculations on Si. When comparing the calculated IVDP's of LA phonon for GaP, InP and InAs and LO phonon for AlAs, AlSb, GaAs, GaSb and InSb with a previous calculation by EPM in rigid approximation, good agreements are found. However, our ab initio pseudopotential results of LA phonon for AlAs, AlSb, GaAs, GaSb and InSb and LO phonon for GaP, InP and InAs are about one order of magnitude smaller than those obtained by EPM calculations, which indicate that the electron redistributions upon the phonon deformations may be important in affecting GAMMA - X intervalley shatterings for these phonon modes when the anions are being displaced. In our calculations the phonon modes of LA and LO at X point have been evaluated in frozen phonon approximation. We have obtained, at the same time, the LAX and LOX phonon frequencies for these materials from total energy calculations. The calculated phonon frequencies agree very well with experimental values for these semiconductors.

INTERVALLEY GAMMA-CHI DEFORMATION POTENTIALS IN III-V ZINCBLENDE SEMICONDUCTORS BY ABINITIO PSEUDOPOTENTIAL CALCULATIONS

Intervalley GAMMA-X deformation-potential constants (IVDP's) have been calculated by use of a first-principles pseudopotential method for the III-V zinc-blende semiconductors AlP, AlAs, AlSb, GaP, GaAs, GaSb, InP, InAs, and InSb. When the calculated IVDP's of LA phonons for GaP, InP, and InAs and of LO phonons for AlAs, AlSb, GaAs, GaSb, and InSb are compared with results of a previous calculation that used the empirical pseudopotential method (EPM) and a rigid-ion approximation, good agreement is found. However, our ab initio pseudopotential results on IVDP's of LA phonons for AlAs, AlSb, GaAs, GaSb, and InSb and of LO phonons for GaP, InP, and InAs are about one order of magnitude smaller than those obtained by use of EPM calculations, indicating that the electron redistribution accompanying crystal-lattice deformation has a significant effect on GAMMA-X intervalley scattering for these phonon modes when the anions are being displaced. In our calculations the LA- and LO-phonon modes at the X point have been evaluated in the frozen-phonon approximation. We have also obtained the LAX- and LOX-phonon frequencies for these materials from total-energy calculations, which agree very well with experimental values for these semiconductors. We have also calculated GAMMA-X hole-phonon scattering matrix elements for the top valence bands in these nine semiconductors, from which the GAMMA-X IVDP's of the top valence bands for the longitudinal phonons and transverse phonons are evaluated, respectively.

1ST PRINCIPLES PSEUDOPOTENTIAL CALCULATIONS OF ZONE BOUNDARY PHONON FREQUENCIES OF III-V-ZINCBLENDE SEMICONDUCTORS

Longitudinal zone boundary X phonon frequencies have been calculated by a first principles pseudopotential method for III-V zincblende semiconductors AlP, AlAs, AlSb, GaP, GaAs, GaSb, InP, InAs and InSb. The phonon frequencies have been evaluated from total energy calculations in the frozen phonon approximation. The calculated phonon frequencies agree very well with the experimental values.

联合检测血清肿瘤四项对恶性肿瘤早期诊断与动态检测的意义

目的　研究血清肿瘤四项标志物联合检测 ,为早期诊断、早期治疗恶性肿瘤提供有价值的临床线索和可靠的依据。方法　对 30 0例患者联合检测血清肿瘤四项标志物 (AFP ,CEA ,γ -GT ,ALP) ,检测结果经临床随访观察 ,验证结论。结果　联合检测血清肿瘤四项标志物对恶性肿瘤早期诊断有效率为 91 3% ,灵敏度为 90 4 % ;对恶性肿瘤患者病情动态监测有效率为 88 0 %。结论　联合检测多种肿瘤标志物明显提高了肿瘤诊断的敏感性。尤其对肝、胆、胰腺肿瘤及肺癌、乳腺癌、胃癌、大肠癌等消化道肿瘤诊断灵敏度高。同时 ,对恶性肿瘤患者的病情进行持续、有效、动态监测与复查 ,利于早期发现肿瘤的复发、转移倾向 ,为早期诊断 ,早期治疗提供有价值的线索和可靠的依据

Synthesis and Metal-ion Binding Properties of New N2S4O2- and N2S5O2-Donor Macrocycles

Synthetic procedures for new mixed-donor macrocycle compounds were reported. The macrocyclic compounds were used in solvent extraction metal picrates such as Ag+, Hg2+, Cd2+, Zn2+, Cu2+, Ni2+, Mn2+, Pb2+, and Co2+. The metal picrate extractions were investigated at 25±0.1°C with the aid of UV-visible spectrometry. It was found that 6,7,9,10,12,13,23,24-octahydro-19H,26Hdibenzo[h,t](1,4,7,13,16,22,10,19) dioxatetrathiadiazasiclotetracosine-20,27(21H,28H)-dione showed selectivity towards Ag+, Hg2+, and Cd2+ among the other metals. The extraction constants (Log Kex) and complex compositions were determined for the Ag+ and Hg2+ complexes for this compound and 9,10,12,13,23,24,26,27,29,30-decahydro-5H,15H-dibenzo-[h,w][1,4,7,13,16,19,25-,10,22] dioxapentathiadiazacycloheptacosine-6,16(7H,17H)-dione.

内蒙古鄂温克族牧民砖茶型氟铝联合中毒调查

目的调查鄂温克族牧民砖茶型氟铝联合中毒病情。方法随机对鄂温克族16~55岁男女均衡62名牧民进行发、血清、尿氟铝水平及血清电解质、生物化学检查,同时检测饮水、牛奶、砖茶、奶茶等氟铝水平,并进行临床检诊和拍摄前臂、骨盆正位X线片,不饮用砖茶的16~55岁男女均衡41名居民为对照组。结果发、血清、尿铝水平牧民组分别为(24.60±9.82)mg/Kg、(0.78±0.25)、(2.84±0.65)mg/L,对照组分别为(12.50±4.55)mg/kg、(0.18±0.11)、(2.09±0.64)mg/L;发、血清、尿氟水平牧民组分别为(1.00±0.52)mg/kg、(0.10±0.10)、(2.74±1.42)mg/L,对照组分别为(1.39±1.34)mg/kg、(0.04±0.03)、(0.87±0.43)mg/L;牧民发、血、尿铝及血、尿氟显著高于对照组(P<0.01)。牧民组血清AST、LDH、HBDH、ALP显著高于对照组(P<0.01)。X线摄片牧民组氟骨症检出率50.00%,关节退行性变检出率61.29%,骨间膜骨化33.87%,疏松型改变检出率29.03%,硬化型改变检出率8.06%;对照组氟骨症检出率2.44%,退行性变检出率34.15%,疏松型改变检出率21.95%,骨间膜骨化检出率2.44%,硬化型改变2.44%,X线氟骨症、关节退行性变、骨间膜骨化牧民组均高于对照组(P<0.01)。结论内蒙古鄂温克族牧民饮茶型氟中毒病情较为严重,其发病机理较为复杂,并导致多组织、器官损伤,可能为砖茶型氟铝联合中毒。

胆总管探查后内置可降解支架并行Ⅰ期缝合的研究

目的　探讨胆总管 (CBD)探查后内置可降解支架并行Ⅰ期缝合在动物实验中的可行性和临床应用价值。方法　用 160只Wistar大鼠模拟CBD探查术 ,将其随机分为两组 :Ⅰ期缝合组 (对照组 ) ,内置可降解支架后行Ⅰ期缝合组 (实验组 )。观察手术前后大鼠碱性磷酸酶 (ALP)、体重、CBD外径的变化 ;记录缝合时间和总手术时间 ;术后观察胆漏的发生率、支架降解情况 ,并取肝组织做病理学检查。结果　两组在缝合时间、总手术时间及胆漏的发生率上差异无显著性 (P >0 .0 5 ) ;术后第 5周时内支架完全降解 ;对照组术后ALP持续性增高 ,实验组术后ALP一过性增高 ;实验组术后体重较对照组明显增加 ;对照组术后CBD较实验组明显扩张 ;对照组术后肝损伤的发生率 (8/15 )显著高于实验组 (1/15 )。结论　CBD探查后内置可降解支架并行Ⅰ期缝合的方法在动物实验中是较为安全有效的。

混合稀土对大鼠肝脏中7种酶的小剂量刺激作用

研究了常乐(用于肥料添加剂的一种混合稀土硝酸盐)对Wistar大鼠肝脏中7种常见酶(ALT,ICD,AST,LAP,ALP,γGT ,CHE)活性的变化,并用电感耦合等离子体质谱(ICP MS)测定了稀土在肝脏中的累积情况。结果表明,稀土对多种酶表现出低剂量激活、高剂量抑制的小剂量刺激作用 (Hormesis),但产生刺激作用的剂量随酶的种类不同而不同 ,并表现出明显的性别差异

四角蛤蜊Mactra veneriformis对镉和汞污染胁迫的生理响应

本论文基于对我国沿海重金属污染加剧以及滩涂贝类资源衰退现象的关注，围绕典型滩涂贝类四角蛤蜊生物化学、细胞生物学和免疫学特征开展研究，调查了渤海湾天津近岸海域四角蛤蜊体内重金属含量和分布规律，以及四角蛤蜊对沉积物重金属的富集能力。室内模拟研究了四角蛤蜊对重金属镉和汞的富集能力及同化机制（assimilation mechanism）差异性，四角蛤蜊各组织对镉和汞的解毒机理和负载能力，以及重金属胁迫对四角蛤蜊血细胞结构和功能的损伤效应。研究结果可望为探讨四角蛤蜊受重金属胁迫细胞水平上的响应机制，揭示重金属污染和贝类资源衰退的关系，建立快速可靠的重金属污染生物标志物指标体系，开展生态健康评估和生境修复等提供科学依据。主要研究结果如下： 1．查明了渤海湾天津近岸四角蛤蜊体内和表层沉积物中Cd、Pb、Cu、Zn、Mn、Cr和Ni七种重金属的含量和周年时空分布特征；表层沉积物中Cd具有较强的污染程度和潜在生态风险，四角蛤蜊对重金属元素Cd和Pb具有较高的富集能力，对环境重金属污染具有很好的指示作用。 2．查明了Cd对四角蛤蜊在24h，48h和96h的半致死浓度（LC50）分别为15.96mg/L、5.15mg/L和2.38mg/L，汞的24h，48h和96h LC50分别为3.71mg/L，0.61mg/L和0.21mg/L。镉和汞对四角蛤蜊的安全质量浓度分别为0.0238mg/L和0.0021mg/L。 3．四角蛤蜊软体部对Cd和Hg的富集能力有显著差异。暴露过程中，四角蛤蜊软体部Cd和Hg的增加量分别为0.12-7.7µg/g和0.002-0.024µg/g，富集率分别为0.3-6.2%和0.11-0.68%，吸收率常数分别为0.07-1.10和0.001-0.005。 4．四角蛤蜊受Cd和Hg胁迫后，外套膜、鳃和肝胰腺的金属硫蛋白（MT）含量均在暴露浓度和胁迫时间上都有极显著变化。组织内MT含量的大小关系为：肝胰腺＞鳃＞外套膜。肝胰腺可以作为双壳贝类MT重金属污染指示研究的目标组织。四角蛤蜊对Cd和Hg不同的解毒机制，导致组织中MT的表达含量和Cd胁迫显著相关，而与Hg胁迫无显著相关性。 5．四角蛤蜊肝胰腺和鳃中抗氧化系统酶、脂质过氧化产物、酸性磷酸酶（ACP）和碱性磷酸酶（ALP）在不同Cd胁迫浓度和暴露时间下有显著差异。与对照组相比，超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPx）的活力和丙二醛（MDA）含量均随着暴露时间的增加而增加，达到一个峰值，然后减低。而ACP和ALP活力则表现出先被抑制，然后升高的变化趋势。肝胰腺中CAT和GPx活力高于鳃，鳃中SOD和MDA含量高于肝胰腺，这种差异与两种组织不同的解毒机理有关。 6．重金属胁迫能够造成四角蛤蜊血细胞超微结构的损伤、溶酶体膜稳定性改变、微核和总畸形核生成，各种损伤均表现出明显的浓度依赖效应。2μg/L Hg暴露14天后，血细胞溶酶体膜中性红保持时间（NRR）、微核生成率（MN）和总畸形核生成率（TNA）与对照组相比没有显著差异。相同暴露时间下，25 μg/L Hg处理组血细胞NRR值低于25 μg/L Cd处理组，而MN和TNA值则相反，说明Hg胁迫对四角蛤蜊血细胞溶酶体膜具有较强的损伤作用。研究发现，NRR、MN和TNA三种指标对于衡量重金属污染对四角蛤蜊血细胞的毒性效应有很好的协同检测作用，可望作为有效的生物标志物在环境监测技术中得以应用。

Modulatory effects of ammonia-N on the immune system of Penaeus japonicus to virulence of white spot syndrome virus

To study response to white spot syndrome virus (WSSV) under ammonia stress, Penaeus japonicus were exposed to 5 mg l(-1) ammonia-N and challenged orally with WSSV (NW). Controls consisted of an ammonia-N-exposed control group (N), a WSSV-challenged positive control group (W), and an untreated control group (control). Immune parameters measured were total haemocyte count (THC), haemocyte phagocytosis, plasma protein content and haemolymph enzymatic activities for prophenoloxidase (proPO), alkaline phosphatase (ALP), and nitric oxide synthase (NOS). THC and plasma protein had downward trends with time in all treatment groups (NW, N, and W) in contrast to the untreated control group (control). The percentage phagocytosis, NOS activity, and ALP and proPO activity of W and NW decreased initially then increased from 6 to 78 h (except for NOS and ALP, from 6 to 54 h) before declining thereafter until the end of the experiment. Compared with untreated controls (control), there was a downward trend for all measured parameters in the treatment groups (N, NW, and W), but the degree was W > NW > N. WSSV was detected at 78 h postchallenge in both W and NW. In conclusion, 5 mg l(-1) ammonia-N reduced the immunocompetence of P japonicus and may have decreased the virulence of WSSV (C) 2004 Elsevier B.V. All rights reserved.