小鼠呼肠孤病毒Ⅲ型标准化血清制备及ELISA检测方法的建立

目的制备标准化小鼠呼肠孤病毒III型（reovirus 3,Reo-3）免疫血清,建立小鼠Reo-3抗体ELISA检测方法。方法使用动物来源的Reo-3病毒感染BALB/c小鼠,获得高效价免疫血清;以BHK-21细胞制备Reo-3病毒抗原,同时制备做正常对照抗原。滴定抗原和标准化小鼠Reo-3血清的最佳工作浓度,进行特异性、敏感性、重复性、稳定性等实验。结果制备18 mL抗Reo-3血清,经检测,IFA效价达1∶640,IEA效价达1∶160;Reo-3抗原和标准化小鼠Reo-3免疫血清最佳工作浓度分别为5μg/mL和1∶2400;该ELISA检测体系Reo-3特异抗原批内变异系数为3.8%,批间变异系数为4.0%;检测灵敏度〉1∶4400;与仙台病毒（SV）、小鼠肝炎病毒（MHV）、小鼠腺病毒（Mad）、小鼠脑脊髓炎病毒（TMEV）、小鼠多瘤病毒（POLY）均无交叉反应。经37℃破坏性试验,2 d稳定性试验相对偏差〈27%。结论本研究制备的Reo-3抗血清达到同批次大量高滴度的水平,可作为检测实验小鼠Reo-3病原的标准化质控血清。本研究建立的ELISA方法重复性、稳定性好,特异性、敏感性强。该体系可用于大、小鼠等实验动物Reo-3抗体的检测。     

Enhanced production of succinic acid by metabolically engineered<Emphasis Type="Italic">Escherichia coli</Emphasis> with amplified activities of malic enzyme and fumarase

Apfl ldhA double mutantEscherichia coli strain NZN111 was used to produce succinic acid by overexpressing theE. coli malic enzyme gene (sfcA). This strain, however, produced a large amount of malic acid as well as succinic acid. After the analyses of the metabolic pathways, thefumB gene encoding the anaerobic fumarase ofE. coli was co-amplified to solve the problem of malic acid accumulation. A plasmid, pTrcMLFu, was constructed, which contains an artificial operon (sfcA-fumB) under the control of the inducibletrc promoter. From the batch culture of recombinantE. coli NZN111 harboring pTrcMLFu, 7 g/L of succinic acid was produced from 20 g/L of glucose, with no accumulation of malic acid. From the metabolic flux analysis the strain was found under reducing power limiting conditions by severe reorientation of metabolic fluxes.     

含铬重组液激活部分缺失金属原子簇的钼铁蛋白的研究

棕色固氮菌（Ａｚｏｔｏｂａｃｔｅｒ ｖｉｎｅｌａｎｄｉｉ）固氮酶钼铁蛋白经邻菲口罗啉和Ｏ２ 处理后,变为部分缺失ＦｅＭｏｃｏ 和Ｐ－ｃｌｕｓｔｅｒ的失活蛋白。与由Ｋ２ＣｒＯ４、高柠檬酸铁、Ｎａ２Ｓ和二硫苏糖醇组成的重组液保温后,处理蛋白对乙炔和质子还原的活性都得以显著恢复;然而,它的吸收光谱和圆二色谱虽有明显恢复,但仍与还原钼铁蛋白有所不同。这表明,激活蛋白中也许存在功能与钼铁蛋白相似,而结构则有所差异的含铬（ＣｒＦｅ）蛋白     

氮磷对污水净化中藻类叶绿素含量的影响

在室内模拟生物净化槽中比较研究了氮磷对污水中藻类叶绿素含量和污水净化的影响。污水中磷含量均为７ｍｇ／Ｌ左右,氮含量分别为７７．４、４４．４、２４．８和１３．５ｍｇ／Ｌ,结果发现ＴＮ／ＴＰ＝７７．４／７．０１ｍｇ／Ｌ组,污水经７ｄ净化,藻类叶绿素含量最高,污水净化效果较好。四个实验组比较,叶绿素含量随ＴＮ／ＴＰ比例的上升而上升,呈显著正相关。     

An 80-kilodalton protein that binds to the pre-S1 domain of hepatitis B virus

It has been suggested that hepatitis B virus (HBV) binds to a receptor on the plasma membrane of human hepatocytes via the pre-S1 domain of the large envelope protein as an initial step in HBV infection. However, the nature of the receptor remains controversial. In an attempt to identify a cell surface receptor for HBV, purified recombinant fusion protein of the pre-S1 domain of HBV with glutathione S-transferase (GST), expressed in Escherichia coli, was used as a ligand. The surface of human hepatocytes or HepG2 cells was biotinylated, and the cell lysate (precleared lysate) which did not bind to GST and glutathione-Sepharose beads was used as a source of receptor molecules. The precleared lysate of the biotinylated cells was incubated with the GST-pre-S1 fusion protein, and the bound proteins were visualized by Western blotting and enhanced chemiluminescence. An approximately 80-kDa protein (p80) was shown to bind specifically to the pre-S1 domain of the fusion protein. The receptor binding assay using serially or internally deleted segments of pre-S1 showed that amino acid residues 12 to 20 and 82 to 90 are essential for the binding of pre-S1 to p80. p80 also bound specifically to the pre-S1 of native HBV particles. Analysis of the tissue and species specificity of p80 expression in several available human primary cultures and cell lines of different tissue origin showed that p80 expression is not restricted to human hepatocytes. Taken together the results suggest that p80 may be a component of the viral entry machinery.     

Effects of macrophage-specific adiponectin expression on lipid metabolism in vivo

Epidemiological studies have associated low circulating levels of the adipokine adiponectin with multiple metabolic disorders, including metabolic syndrome, obesity, insulin resistance, type II diabetes, and cardiovascular disease. Recently, we reported that adiponectin selectively overexpressed in mouse macrophages can improve insulin sensitivity and protect against inflammation and atherosclerosis. To further investigate the role of adiponectin and macrophages on lipid and lipometabolism in vivo, we engineered the expression of adiponectin in mouse macrophages (Ad-TG mice) and examined effects on plasma lipoproteins and on the expression levels of genes involved in lipoprotein metabolism in tissues. Compared with the wild-type (WT) mice, Ad-TG mice exhibited significantly lower levels of plasma total cholesterol (-21%, P < 0.05) due to significantly decreased LDL (-34%, P < 0.05) and VLDL (-32%, P < 0.05) cholesterol concentrations together with a significant increase in HDL cholesterol (+41%, P < 0.05). Further studies investigating potential mechanisms responsible for the change in lipoprotein cholesterol profile revealed that adiponectin-producing macrophages altered expression of key genes in liver tissue, including apoA1, apoB, apoE, the LDL receptor, (P < 0.05), and ATP-binding cassette G1 (P < 0.01). In addition, Ad-TG mice also exhibited higher total and high-molecular-weight adipnection levels in plasma and increased expression of the anti-inflammatory cytokine IL-10 as well as a decrease in the proinflammatory cytokine IL-6 in adipose tissue. These results indicate that macrophages engineered to produce adiponectin can influence in vivo gene expression in adipose tissue in a manner that reduces inflammation and macrophage infiltration and in liver tissue in a manner that alters the circulating lipoprotein profile, resulting in a decrease in VLDL and LDL and an increase in HDL cholesterol. The data support further study addressing the use of genetically manipulated macrophages as a novel therapeutic approach for treatment of cardiometabolic disease.     

Dynamic distribution of spindlin in nucleoli, nucleoplasm and spindle from primary oocytes to mature eggs and its critical function for oocyte-to-embryo transition in gibel carp

Spindlin (Spin) was thought as a maternal-effect factor associated with meiotic spindle. Its role for the oocyte-to-embryo transition was suggested in mouse, but its direct evidence for the function had been not obtained in other vertebrates. In this study, we used the CagSpin-specific antibody to investigate CagSpin expression pattern and distribution during oogenesis of gibel carp (Carassius auratus gibelio). First, the oocyte-specific expression pattern and dynamic distribution was revealed in nucleoli, nucleoplasm, and spindle from primary oocytes to mature eggs by immunofluorescence localization. In primary oocytes and growth stage oocytes, CagSpin accumulates in nucleoli in increasing numbers along with the oocyte growth, and its disassembly occurs in vitellogenic oocytes, which implicates that CagSpin may be a major component of a large number of nucleoli in fish growth oocytes. Then, co-localization of CagSpin and β-tubulin was revealed in meiotic spindle of mature egg, indicating that CagSpin is one spindle-associated factor. Moreover, microinjection of CagSpin-specific antibody into the fertilized eggs blocked the first cleavage, and found that the CagSpin depletion resulted in spindle assembly disturbance. Thereby, our study provided the first direct evidence for the critical oocyte-to-embryo transition function of Spin in vertebrates, and confirmed that Spin is one important maternal-effect factor that participates in oocyte growth, oocyte maturation, and oocyte-to-embryo transition.     

种子老化诱导小麦染色体畸变及大麦醇溶蛋白带型频率变化的研究

通过时20份小麦种质发芽率和根尖细胞染色体畸变的测定,结果表明同一品种贮藏于中期库的低发芽率种质,其染色体畸变率明显高于贮藏于长期库的高发芽率种质.从总体上看,种子根尖细胞染色体畸变率与发芽率呈负显著相关,而与贮藏年限无显著相关.但对长期库和中期库的种质分别进行统计分析,种子根尖染色体畸变率与发芽率和贮藏年限均无显著相关.对大麦地方品种"普乃干木"的醇溶蛋白分析表明,该品种有4种biotype,随着发芽率的下降,其群体中的4种bio-type的频率发生了变化,当发芽率从95%降到34%时,其中一种biotype出现了消失,而有一种biotype频率从83%上升至95%,此结果表明,种子生活力下降和不同biotype种子存活能力存在差异,可能导致了异质种质材料遗传选择和漂变的发生.     

杉木与固氮和非固氮树种混交对林地土壤质量和土壤水化学的影响

第 1代人工杉木林皆伐后 ,3种不同的经营模式 ,即连载杉木纯林、杉木与固 N阔叶树混交林和杉木与非固 N阔叶树混交林 ,对林地土壤质量和土壤水化学的影响进行了比较。结果表明 ,在杉树与阔叶树混交经营模式下 ,土壤养分含量增加 ,物理性状改善 ,土壤生物活性提高 ,微生物商 (Cmic:Corg)上升 ,代谢商 (q CO2 )稍有下降 ,但杉木与固 N树种的混交对土壤质量的改善效果比杉木与非固 N树种混交好 ;相反 ,杉木连载只能导致林地土壤质量的逐渐恶化 ;土壤溶液中 ,主要来自于大气中的一些离子浓度 ,如 SO2 - 4,Cl- ,Na 和 Mg2 ,在杉木纯林中显著高于混交林 ,而主要受系统内影响较大的一些离子 ,如 K 和NH 4,NO- 3,在经营模式间变异较小 ;H 和 Al3 浓度也是杉木纯林比混交林高。另外 ,研究结果还表明 ,总有机 C、CEC和微生物 C与其它土壤理化性质与生物学性质之间存在着较好的相关性 ,所以可以将总有机 C、CEC和微生物 C作为红黄壤地区亚热带人工林土壤质量的指示指标     

An Imported Case of Cystic Echinococcosis in the Liver

A 25-year-old Uzbek male presented with right upper abdominal pain for 20 days. On radiologic studies, a huge cystic mass was noticed in the right liver which was suspected as parasitic. The patient received right hepatic segmentectomy (segment 7), and the surgically resected mass was confirmed as cystic echinococcosis (CE), measuring 10.5 cm in its diameter. The inner surface of the cyst was bile-stained. The patient was discharged on the 8th hospital day, and was rechecked 6 months after the surgical intervention without any evidence of recurrence. The present report describes findings of an imported case of CE which represented ultrasound images of the ''ball of wool''.