Shikonin reverses HGF-induced resistance to gefitinib in lung cancer cells HCC827

AIM: To investigate the effect of shikonin on reversing hepatocyte growth factor(HGF)-induced resistance to gefitinib in lung cancer HCC827 cells, and to explore its possible mechanisms.METHODS: The gefitinib-resistant HCC827 cells induced by HGF were treated with shikonin and gefitinibthe alone or in combination. The inhibition rates of cell viability were determined by MTT assay. The invasive ability of HCC827 cells with HGF-induced resistance to gefitinib was determined by Transwell assay. The protein levels of epithelial-mesenchymal transition (EMT) and related signaling pathway in the HCC827 cells were detected by Western blot.RESULTS: The results of MTT assay showed that the cell activity of HCC827 cells was significantly inhibited by shikonin in a dose dependent manner. The IC₅₀ of shikonin in HCC827 cells was 3.06 μmol/L. And the IC₅₀ of gefitinib in HCC827 cells was 0.51 μmol/L. Under the condition of combined treatment with shikonin and gefitinib in the presence of HGF (20 μg/L), the IC₅₀ of gefitinib was 7.36 μmol/L, significantly lower than that treated with gefitinib alone (P<0.01), so did the result of the cell migration (P<0.01). HGF induced EMT, while shikonin reversed this effect. The protein expression level of p-AKT was significantly up-regulated by HGF, while markedly down-regulated treatment with shikonin and gefitinib compared with gefitinib alone (P<0.01).CONCLUSION: Shikonin reverses HGF-induced resistance to gefitinib in lung cancer HCC827 cells, and the mechanism may be likely related to the preventon of EMT and the inhibition of HGF-induced activation of p-AKT signaling pathway.     

鸡流行性腹泻病原的分离鉴定及特性研究

用胰蛋白酶处理发病鸡的粪便和肠内容物，接种Marc145细胞，盲传数代后，从山东不同地区发生流行性腹泻的鸡群中分离到轮状病毒，并对分离的轮状病毒的生物学特性和理化特性进行了部分研究。结果表明病毒粒子的形态呈车轮状、大小为70-80nm；其病毒基因组的电泳图谱为5：1：3：2；病毒在Marc145细胞上传到第9代时的TCID50为10^-4.62／0．1mL；该分离株病毒对氯仿、乙醚有抵抗力；对pH3．0处理60min稳定；50℃ 30min能使其感染力下降10^2；1mol／L MgCl2不能增强其对50℃ 60min的抵抗力。动物回归试验中接种两周龄SPF鸡，24h后陆续发病，表现为持续性水样腹泻，与自然发病相同；剖检可见病鸡脱水、小肠内有大量的液体和气泡、肠粘膜变薄；组织学变化为肠绒毛上皮坏死、脱落，绒毛平均长度减少而隐窝深度增加，固有层中淋巴细胞浸润。其临床症状及病理组织学变化与自然发病相同。因此确定发生在山东鸡流行件腹泻的病原为轮状病毒．     

法氏囊活性肽在大肠杆菌中的克隆与表达

法氏囊活性肽（Bursin,BS)作为一种免疫增强剂，临床应用广泛，为了大量制备BS，按大肠杆菌惯用密码子合成BS基因。并用色氨酸将5个BS基因串联，克隆于质粒pU57的SmaI位点，经测序证明序列正确后，以PCR扩增，克隆于表达载体pBV220的EcoRI,HinDⅢ位点，温敏诱导表达，其表达水平占全菌总蛋白的13.4%。     

多重PCR检测猪瘟病毒、猪细小病毒、猪伪狂犬病病毒

根据猪瘟病毒(CSFV)、猪细小病毒(PPV)和猪伪狂犬病病毒(PRV)3种病原体的基因保守序列，分别设计了与CSFV的E2基因、PPV的VP2基因和PRV的gⅡ基因序列互补的3对引物。用这3对引物对人为混合的样品中的PPV和PRV的DNA模板及CSFV反转录后的cDNA模板进行多重PCR扩增和多重PCR反应奈件的优化，结果同时得到3务与试验设计相符的288bp(CSFV)、575bp(PPV)700bp(PRV)特异性条带；敏感性检测结果表明，该多重PCR可以检测到14．5ng／L的CSFV、27．1pg／L的PPV、31．4ng／L的PRV的核酸模板。     

武功山国家森林公园的自然生态化规划设计实践

森林是重要的自然资源，在开发方式上应十分慎重。以自然生态化规划设计思想作为指导思想，进行了武功山国家森林公园的规划和设计，进行了森林公园可持续规划设计的探索。     

腐蚀管道剩余寿命的性能衰减模型研究

管道在长期的运行过程中，由于介质和土壤的腐蚀性，管材的微塑性变形、结构转变和渗氢等。不仅会使管道出现各种腐蚀损伤，而且还存在着老化效应和氢化作用，导致管材的脆变并改变材料的塑性，使机械性质参数发生衰退，从管材性能衰减和腐蚀损伤角度研究了管道的剩余寿命，提出了根据结构承载能力的极限值确定塑性变形时工作寿命的方法，为了确定剩余使用寿命，对剪切损伤率和屈服性能比等概念进行定义，在掌握管材金属剪切损害率和屈服性能比的时间特性基础上，建立了剩余寿命计算模型。并给出了算例。     

聚丁二炔链中的光吸收和光感应键结构改变

1985年Koda等人发现聚丁二炔在光的照射下,会发生从兰色到红色的变化;掺杂不同时,其光吸收出现了一系列新的特征,并在有机光导,非线性光学层,电子束复印材料等方面得到了应用.本文首次提出一个包括电子—电子,电子—声子相互作用的模型,成功地解释了有关实验现象.     

不同方法构建磺胺二甲氧嘧啶免疫抗原的比较研究

为探讨合成磺胺二甲氧嘧啶(SDM)免疫抗原的有效途径,采用重氮偶合法和戊二醛法分别合成SDM免疫抗原,并对其结构特征、光谱特征和SDM结合比进行比较.结果表明,在合成SDM-BSA中采用重氮偶合法比戊二醛法好.     

Genotype evaluation,muscle histopathology and ultrastructure in a Duchenne muscular dystrophy animal model

AIM: To evaluate the genotype , muscle histopathology and ultrastructure in dko mice. METHODS: Dystrophin/Utrophin-deficient double knockouts (dko) mice were obtained from university of Oxford, UK. Genotype of filial generation of heterozygote was evaluated by PCR-SSP. HE staining and fluorescent immunohistochemistry by SABC-Cy3 were used to detect striated muscle of dko mouse, and the muscle ultrastructure was observed by transmission electron microscope(TEM). RESULTS: In 112 filial generation mice, there were 28 mdx (25.0%), 26 dko (23.2%) and 58 heterozygote (51.8%), which coincided with the law of Mendelian inheritance. HE staining showed that the myocytes were not very uniform, there were phenomenon of round outline, centrally nucleated fibers, widening interspace, inflammatory cell infiltration and connective tissue proliferation in dko mice. There were no any immunofluorescent expression of dystrophin and utrophin in sarcolemma in dko mice. TEM showed sarcolemma breakage, separation and edema, and loose myofibril texture, inflammatory cell infiltration and connective tissue proliferation in dko mice. CONCLUSION: PCR-SSP is a very quick and accurate way for genotype evaluation of filial generation. The pathophysiology of dko mouse was very similar to Duchenne muscular dystrophy (DMD), and dko mouse is an ideal animal model for study of DMD clinical therapy.     

中国上市公司多元化经营绩效的实证分析

通过随机抽样,计算我国57家上市公司1998年-2002年的平均盈利指标和风险指标,根据各公司多元化熵值的大小对样本总体进行分组,分析多元化程度对企业绩效的影响.研究发现1.多元化程度越高,公司的平均净资产收益率越高;2.多元化经营降低了公司收益率的波动水平;3.资产规模越小的公司越倾向于选择多元化经营.