人IFN-g体外释放检测法的建立及其在结核病诊断中的应用

本研究旨在建立人IFN-g体外释放检测法, 用于人结核病的特异性诊断。克隆表达了人IFN-g基因, 利用纯化的重组IFN-g免疫小鼠, 获得两株高效价的单克隆抗体。用所获得的单克隆抗体及兔抗IFN-g多克隆抗体建立了检测人IFN-g的夹心ELISA, 检测灵敏度达到31.25 pg/mL。采集111位结核病阳性病人与292位临床健康对照者肝素抗凝全血,利用结核菌特异性抗原ESAT-6/CFP-10融合蛋白体外刺激外周血淋巴细胞释放IFN-g, 用所建立的夹心ELISA及商品化试剂盒平行检测所有样本, 结果表明两种方法的检测结果相符。结核患者的检测灵敏度为95.5%, 健康对照的阳性检出率为16.7%, 患者与健康对照的阳性检出率差异极显著(P<0.01), 证实所建立的方法灵敏度与特异性均很高, 具有良好的应用前景。     

Microbial metabolism of 1-aminoanthracene by Beauveria bassiana

The carcinogen and mutagen, 1-aminoanthracene, was efficiently metabolized by the fungal strain Beauveria bassiana ATCC 7159 to yield three new metabolites identified as 1-acetamido-5-[(4′-O-methyl-β-d-glucopyranosyl)oxy]anthracene, 1-acetamido-8-[(4′-O-methyl-β-d-glucopyranosyl)oxy]anthraquinone, and 1-acetamido-6-[(4′-O-methyl-β-d-glucopyranosyl)oxy]anthraquinone, together with 1-acetamidoanthracene and 1-acetamidoanthraquinone. Formation of these metabolites suggests that the metabolic pathways of 1-aminoanthracene in B. bassiana ATCC 7159 involve acetylation, oxidation, hydroxylation, and O-methylglucosylation.     

Variations of enzyme activities in the haemocytes of scallop Chlamys farreri after infection with the acute virus necrobiotic virus (AVNV)

Acute virus necrobiotic virus (AVNV) is one of the main pathogens for large scale mortality of Chinese scallop Chlamys farreri. In this paper, C. farreri were infected by different dilutions of AVNV supernatant (5(0), 5(-1), 5(-2), 5(-3), 5(-4), 5(-5), 5(-6), 5(-7), respectively), and dead individuals were counted every day for 15 days. Samples from groups of 5(-3) and 5(-5) were taken every day till 15 days and the activities of acid phosphatase (ACP), alkaline phosphatase (ALP), superoxide dismutase (SOD), myeloperoxidase (MPO), phenoloxidase (PO), peroxidase (POD) and catalase (CAT) in haemocytes were measured. The results of virus challenge showed that survival rates of scallops in groups of 5(0) and 5(-1) decreased sharply after the first day and died out completely on the 5th and 4th day, respectively. In other groups (5(-2), 5(-3), 5(-4), 5(-5), 5(-6) and 5(-7)), survival rates decreased gradually till 6 or 7 days, then kept steady till 15 days, and they were dose-dependent, increasing from 12% to 80% as the dose decreased from 5(-2) to 5(-7) viral supernatant. In the control group, survival rate was 88%. Enzyme activities for groups of 5(-3) and 5(-5) illustrated that activities of ACP, SOD, MPO, PO in groups of 5(-3) and 5(-5) were significantly higher than the control group in the first 9 or 10 days, and went back to the control group levels gradually after 10 days. Moreover, their activities in group of 5(-3) varied more than that in the group of 5(-5), especially activities of MPO, PO. Differently, the activities of POD and CAT were reduced or induced by virus infection and showed no regular trends in the experiments. The activity of ALP was not detected.     

城市污水处理系统的菌群分析和除氮基因工程出发菌的选择

目的:对脱氮污水处理工艺的活性污泥的菌群组成进行分析,以期获得适合于脱氮基因工程改良的出发菌.方法:首先采用平板稀释法对活性污泥进行菌落计数,并对分离到的菌落进行详细的生化鉴定,对其中的优势菌-假单胞菌进行脱氮能力测定,并分析其对常作为筛选标志的抗生素的药物敏感性.结果:发现在采用该工艺的活性污泥中,优势菌为假单胞菌、肠杆菌、莫拉菌和不动杆菌,分别占总菌数的23%、16%、16%和12%.根据菌群分析的结果,从中选择了两株耐药性弱、脱氮能力强的菌作为基因改良的出发菌.结论:本研究阐明了活性污泥的菌群构成,获得了两株适合基因工程改良的菌株,为日后脱氮基因工程菌的构建奠定基础.     

多氯联苯对剑尾鱼超氧化物岐化酶活性的影响

目的　研究多氯联苯 (PCBs)暴露对剑尾鱼 (Xiphophorushelleri)超氧化物歧化酶 (SOD)活性的影响 ,探讨剑尾鱼器官组织内SOD活性变化作为环境风险评价 (ERA)的有效生物学标记的可行性。方法　测定了PCBs对剑尾鱼 30d的半致死浓度 (LC50 ) ;使用浸浴法以 0、2、5 0 μg L三个PCBs浓度为剑尾鱼染毒 ,定量测定了 72h内肝、鳃及卵巢组织中的SOD的活性变化。结果　PCBs对剑尾鱼的 30dLC50 为 1 0 5 80 μg L ;PCBs对剑尾鱼肝脏和卵巢的SOD活性有明显 (P <0 0 1 )的影响。在最初染毒的 1 2h内 ,SOD活性略有上升 ,但随暴露时间延长 ,浓度增加 ,肝和卵巢的SOD活性均呈下降趋势。此外 ,结果还显示肝组织的SOD活性较高于卵巢的SOD活性 ,表明不同器官组织的SOD活性对PCBs胁迫的敏感性存在一定的差异。实验中鳃组织SOD活性在PCBs暴露后其变化不明显 (P >0 0 5 )。结论　表明剑尾鱼肝细胞SOD活性可作为环境风险评价 (ERA)的有效生物学标记     

Nutrient removal from slaughterhouse wastewater in an intermittently aerated sequencing batch reactor

The performance of a 10 L sequencing batch reactor (SBR) treating slaughterhouse wastewater was examined at ambient temperature. The influent wastewater comprised 4672+/-952 mg chemical oxygen demand (COD)/L, 356+/-46 mg total nitrogen (TN)/L and 29+/-10 mg total phosphorus (TP)/L. The duration of a complete cycle was 8 h and comprised four phases: fill (7 min), react (393 min), settle (30 min) and draw/idle (50 min). During the react phase, the reactor was intermittently aerated with an air supply of 0.8L/min four times at 50-min intervals, 50 min each time. At an influent organic loading rate of 1.2g COD/(Ld), average effluent concentrations of COD, TN and TP were 150 mg/L, 15 mg/L and 0.8 mg/L, respectively. This represented COD, TN and TP removals of 96%, 96% and 99%, respectively. Phase studies show that biological phosphorus uptake occurred in the first aeration period and nitrogen removal took place in the following reaction time by means of partial nitrification and denitrification. The nitrogen balance analysis indicates that denitrification and biomass synthesis contributed to 66% and 34% of TN removed, respectively.     

Ornamental Expression of Red Fluorescent Protein in Transgenic Founders of White Skirt Tetra (<Emphasis Type="Italic">Gymnocorymbus ternetzi</Emphasis>)

Although the transgenic technology has been successfully used to generate fluorescent zebrafish and medaka for ornamental purposes, the practicability of the technology has not been demonstrated in other ornamental fish species. In the present study, we have tested the transgenic technology in a bona fide ornamental fish species, the white skirt tetra (Gymnocorymbus ternetzi). First, its embryonic development was briefly described. Second, we successfully introduced an rfp (red fluorescent protein) gene construct driven by a strong muscle-specific mylz2 promoter from the zebrafish into the white skirt tetra and demonstrated muscle-specific expression of the RFP reporter protein. Importantly, the vivid red fluorescent color was prominently visible in adult transgenic founders under the normal daylight, like the currently marketed red fluorescent transgenic zebrafish. Thus, our current study demonstrated the feasibility of using the well-characterized zebrafish mylz2 promoters to produce useful fluorescent ornamental fish in other fish species by the transgenic technology.     

Comparison of Gene Expression Profiles of Fenneropenaeus chinensis Challenged with WSSV and Vibrio

Microarray technique was used to analyze the gene expression profiles of shrimp when they were challenged by WSSV and heat-inactivated Vibrio anguillarum, respectively. At 6 h post challenge (HPC), a total of 806 clones showed differential expression profile in WSSV-challenged samples, but not in Vibrio-challenged samples. The genes coding energy metabolism enzyme and structure protein were the most downregulated elements in 6 h post WSSV-challenged (HPC-WSSV) tissues. However, a total of 155 clones showed differential expression in the Vibrio-challenged samples, but not in WSSV-challenged samples. Serine-type endopeptidase and lysosome-related genes were the most upregulated elements in tissues 6 h post Vibrio challenge (HPC-Vibrio). Totally, 188 clones showed differential expression in both 6 and 12 HPC-WSSV and HPC-Vibrio samples. Most of the differentially expressed genes (185/188) were downregulated in the samples of 12 HPC-WSSV, whereas upregulated in the samples at 6 and 12 HPC-Vibrio and 6 HPC-WSSV. The expression profiles of three differentially expressed genes identified in microarray hybridization were analyzed in hemocytes, lymphoid organ, and hepatopancreas of shrimp challenged by WSSV or Vibrio through real-time PCR. The results further confirmed the microarray hybridization results. The data will provide great help for us in understanding the immune mechanism of shrimp responding to WSSV or Vibrio. Wang and Li contributed equally to this work.     

Accurate methods of DNA extraction and PCR-based genotyping for single scallop embryos/larvae long preserved in ethanol

Marine scallops are sessile as adult but have a long planktonic larval phase showing great possibility to migrate in marine realm lacking of obvious barriers. Genetic analysis of scallop embryos/larvae based on molecular markers is very essential to clarify the spatial and temporal gene flow and the unique population and community structure. However, the technical challenges, such as single embryos/larvae isolation and low quantity and poor quality of DNA extracted, make genotyping for a single embryo/larva long preserved in ethanol to be a really difficult task. In this study, we analysed the factors that might affect the DNA quantity and quality for simple sequence repeat‐based genotyping for single embryos/larvae. Based on the factors analysed, we developed a LoTEPA buffer‐based method, of which the accuracy, stability and reproducibility were evaluated by controlled inter‐ and intraspecies and self‐fertilized scallop families. The genotyping results showed the high success rate of more than 90% in total for embryos/larvae preserved in ethanol for 1–5 years. Furthermore, the successful genotyping for the larvae sampled from a natural habitat well demonstrated the potential use of this method in practical ecological analysis.