灵芝免疫调节蛋白(Lz-8)在毕赤酵母中的表达及其免疫活性鉴定

真菌免疫调节蛋白家族(Fungi immunoregulatory proteins,FIPs)各成员所具有的免疫调节和抗肿瘤活性已被广泛研究。本研究利用毕赤酵母表达系统对其成员Lz-8进行了重组表达。以毕赤酵母突变株GS115为表达宿主细胞,PCR和DNA测序结果均显示Lz-8的cDNA已被成功地整合入酵母基因组。聚丙烯酰胺凝胶电泳(SDS-PAGE)、激光解析飞行时间质谱(MALDI-TOF-MS)和免疫学实验均被用于重组表达蛋白的检测。实验结果表明Lz-8在毕赤酵母表达系统中得到成功表达,在SDS-PAGE中可观察到分子量为14000D的单一条带,MALDI-TOF-MS的实验结果显示rLz-8的分子量为12722D。在相关的免疫学实验中,rLz-8可引起绵羊血红细胞凝集,但对人血4种血型的红细胞并没有凝集作用,rlz-8还可诱导巨噬细胞吞噬作用,均与其他报道中的实验结果吻合。以上结果表明,本实验已成功地利用毕赤酵母表达系统对Lz-8进行重组表达。     

TSG101 promotes the proliferation,migration and invasion of hepatocellular carcinoma cells by regulating the PEG10

The tumour susceptibility gene 101 (TSG101) is reported to play important roles in the development and progression of several human cancers. However, its potential roles and underlined mechanisms in human hepatocellular carcinoma (HCC) are still needed to be further clarified. In the present study, we reported that knock down of TSG101 suppressed the proliferation, migration and invasion of HCC cells, while overexpression of TSG101 facilitated them. Molecularly, the results revealed that knock down of TSG101 significantly decreased the cell cycle related regulatory factor p53 and p21. In another point, knock down of TSG101 also obviously decreased the level of metallopeptidase inhibitor TIMP1 (Tissue inhibitors of metalloproteinases 1), which results in inhibition of MMP2, MMP7 and MMP9. In contrast, overexpression of TSG101 had opposite effects. The iTRAQ proteomics analysis identified that oncogenic protein PEG10 (Paternally expressed gene 10) might be a potential downstream target of TSG101. Further investigation showed that TSG101 interacted with PEG10 and protected it from proteasomal degradation thereby regulating the expression of p53, p21 and MMPs. Finally, we found that both TSG101 and PEG10 proteins are up‐regulated and presented a direct correlation in HCC patients. In conclusion, these results suggest that TSG101 is up‐regulated in human HCC patients, which may accelerate the proliferation, migration and invasion of HCC cells through regulating PEG10.     

ZNRF3 acts as a tumour suppressor by the Wnt signalling pathway in human gastric adenocarcinoma

E3 ubiquitin ligases regulate a variety of biological processes through the ubiquitin–proteasome system, together with ubiquitin activating enzyme E1 and ubiquitin-conjugating enzyme E2. Previous studies have demonstrated that zinc and ring finger 3 (ZNRF3), which belongs to the E3 ubiquitin ligases family is involved in the Wnt signalling pathway, which plays an important role in causing cancer. However, the expression and function of ZNRF3 in human gastric adenocarcinoma still remains unclear. Immunohistochemical and western blot analysis showed a significant down-regulation of ZNRF3 protein in gastric adenocarcinoma tissues compared with adjacent normal gastric tissues. In addition, there was a correlation between the down-regulation of ZNRF3 and poor tissue differentiation in gastric adenocarcinoma. To investigate the potential function of ZNRF3 in cell proliferation and apoptosis, a gastric cell line SGC7901 was employed. The over-expression of wild-type ZNRF3, which was accomplished by the transient transfection of recombinant pEGFP-ZNRF3 (or empty plasmids as control) into the cell line SGC7901, was confirmed by western blot analysis. Flow-cytometry-based and Cell Counting Kit-8 assays showed that over-expression of wt ZNRF3 induced apoptosis and suppressed proliferation. ZNRF3-overexpressing gastric cells displayed partly attenuated protein levels of beta-catenin and TCF-4 compared with those transfected with the empty plasmid. Our study demonstrates a novel gastric adenocarcinoma suppressor and reveals that ZNRF3 inhibits gastric cancer cell growth and promotes the cell apoptosis by affecting the Wnt/beta-catenin/TCF signalling pathway.     

Silicon-mediated alleviation of Mn toxicity in Cucumis sativus in relation to activities of superoxide dismutase and ascorbate peroxidase

The effects of exogenous silicon (Si) on plant growth, activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and catalase, and concentrations of ascorbate and glutathione were investigated in cucumber (Cucumis sativus L.) plants treated with excess manganese (Mn) (600 microM). Compared with the treatment of normal Mn (10 microM), excess Mn significantly increased H2O2 concentration and lipid peroxidation indicated by accumulation of thiobarbituric acid reactive substances. The leaves showed apparent symptoms of Mn toxicity and the plant growth was significantly inhibited by excess Mn. The addition of Si significantly decreased lipid peroxidation caused by excess Mn, inhibited the appearance of Mn toxicity symptoms, and improved plant growth. This alleviation of Mn toxicity by Si was related to a significant increase in the activities of SOD, APX, DHAR and GR and the concentrations of ascorbate and glutathione.     

中华鲟YY肽基因电子克隆表达与生物信息学分析

以日本鳗鲡的PYY基因cDNA序列为信息探针,搜索中华鲟EST文库,得到中华鲟PYY的EST序列,经过生物信息学分析。结果表明,此cDNA序列包含完整的开放读码框,所编码的蛋白质包含97个氨基酸,前28个氨基酸为信号肽,分子量为11．03kD,理论等电点为5．54。该蛋白序列和牙鲆、河豚以及斑马鱼的PYY肽同源性较高。其中36个氨基酸构成PAH结构域,由一个α螺旋结构和一个无规则卷曲区组成。包含中华鲟PYY在内的139条真核生物神经肽Y蛋白质序列比对结果显示,有6个氨基酸位点高度保守,其中有5个氨基酸位于α螺旋上,另外1个脯氨酸位于无规则卷曲区,提示α螺旋也可能起到重要作用。     

Association of basal serum testosterone levels with ovarian response and in vitro fertilization outcome

Background  

To evaluate basal testosterone (T) levels during follicular phase of the menstrual cycle as a predictor for ovarian response and in vitro fertilization (IVF) outcome.     

新生儿乙型肝炎血源疫苗的免疫策略和成本效益分析

比较了6种新生儿血源乙型肝炎疫苗的免疫方案,其中一种只免疫HBsAg阳性母亲的婴儿效果最差,群体有效率仅17.79％,成本效益比值亦低。其它5种方案都是对全部新生儿进行免疫,效果均好。这5种方案是10μg与3μg两种剂量的不同组合,其中用10μgg3JI疫者成本效益比值最高,有效率为78.61％。HBsAg阳性母亲新生儿按30μg×3,而阴性母亲婴儿首剂用30μg,第2、3剂各10μg,有效率高达87.47％,但成本效益比值最低。应根据我国经济发展的水平与群众的承受能力,对各地区选用不同的新生儿免疫方案。     

Glutamate transporter type 3 knockout leads to decreased heart rate possibly via parasympathetic mechanism

Parasympathetic tone is a dominant neural regulator for basal heart rate. Glutamate transporters (EAAT) via their glutamate uptake functions regulate glutamate neurotransmission in the central nervous system. We showed that EAAT type 3 (EAAT3) knockout mice had a slower heart rate than wild-type mice when they were anesthetized. We design this study to determine whether non-anesthetized EAAT3 knockout mice have a slower heart rate and, if so, what may be the mechanism for this effect. Young adult EAAT3 knockout mice had slower heart rates than those of their littermate wild-type mice no matter whether they were awake or anesthetized. This difference was abolished by atropine, a parasympatholytic drug. Carbamylcholine chloride, a parasympathomimetic drug, equally effectively reduced the heart rates of wild-type and EAAT3 knockout mice. Positive immunostaining for EAAT3 was found in the area of nuclei deriving fibers for vagus nerve. There was no positive staining for the EAATs in the sinoatrial node. These results suggest that EAAT3 knockout mice have a slower heart rate at rest. This effect may be caused by an increased parasympathetic tone possibly due to increased glutamate neurotransmission in the central nervous system. These findings indicate that regulation of heart rate, a vital sign, is one of the EAAT biological functions.