Choroid plexus carcinoma cells in the cerebrospinal fluid of a Staffordshire Bull Terrier

An 11-year-old female intact Staffordshire Bull Terrier was referred to the Queen's Veterinary School Hospital at the University of Cambridge with sudden onset of episodic behavioral changes, a mammary mass, and papilledema in the right eye. On physical examination the dog appeared depressed and had a head tilt to the right with anisocoria. Using magnetic resonance imaging, a broad-based lesion that obliterated the fourth ventricle was detected in the right brainstem. There was no evidence of pulmonary metastasis. Cerebrospinal fluid (CSF) was then obtained; fluid analysis showed an increased cell count (165 cells/μL, reference interval 0-7 cells/μL) and total protein (0.30 g/L, reference value <0.25 g/L). Cytologic evaluation revealed a population of atypical epithelial cells arranged in cohesive rafts and characterized by moderate to occasionally marked anisocytosis and anisokaryosis. The appearance was highly suspicious of a malignant epithelial neoplasm. The dog was euthanized and on postmortem examination an asymmetrical nonencapsulated cerebellar mass was found within the choroid plexus of the fourth ventricle with local extension into the cerebellopontine angle. Histologic sections of the cerebellar mass contained arborizing papillary structures covered by a single layer of atypical epithelial cells that showed local infiltration into the adjacent neuropil. The diagnosis was choroid plexus carcinoma. The atypical epithelial cells were negative for pancytokeratin and strongly positive for vimentin. The finding of clusters of choroid plexus epithelial cells in the CSF demonstrates the value of utilizing a relatively noninvasive diagnostic technique for diagnosis of choroid plexus tumors.     

Ganoderma lucidum extract protects rat cerebral cortical neurons from hypoxia/reoxygenation injury in vitro

AIM: To investigate the neuroprotective effect of Ganoderma lucidum extract (GLE) in an in vitro model of primary cultured neurons with oxygen and glucose deprivation (OGD). METHODS: Neuronal injury was induced by oxygen and glucose deprivation/reoxygenation (OGD/R). The neuronal injury and viability were determined by LDH leakage and XTT assay at 0 h,3 h,6 h,12 h,24 h,48 h and 72 h after OGD/R. Neuronal apoptosis was detected by flow cytometry (FCM). The expression of apoptosis-related proteins was analyzed by Western blotting.RESULTS: The viability of the neurons increased with exposure to GLE (0.1 mg/L,1 mg/L and 10 mg/L)after OGD/R. The LDH releases were also significantly reduced. GLE significantly inhibited OGD/R-induced apoptosis of cultured rat cortical neurons in a concentration-dependent and time-dependent manner(P<0.05). GLE at concentrations of 0.1 mg/L,1 mg/L and 10 mg/L inhibited the expression of caspase-3 and caspase-8 proenzyme. Additionally,GLE at concentration of 10 mg/L suppressed the expression of caspase-9 proenzyme.CONCLUSION: Our findings provide the evidence that the GLE has neuroprotective effect on cerebral ischemia. The mechanisms are related to the inhibition of caspase-3,-8 and-9 activations. GLE may be a novel and effective reagent for treating ischemic stroke.     

近红外量子点表皮生长因子单克隆抗体荧光探针对金黄地鼠颊黏膜癌变过程的动态可视化荧光成像研究

目的 利用近红外量子点表皮生长因子单克隆抗体荧光探针对由9,10-二甲基1,2苯并蒽（9,10-dimethylen-1,2-benzanthracene,DMBA）诱导建立的金黄地鼠颊黏膜癌变过程进行动态可视化荧光成像研究。方法 （1）用0.5%DMBA丙酮液诱导建立金黄地鼠颊黏膜癌变各期模型;（2）将水溶性近红外荧光量子点（QD800）表面功能性的修饰藕连表皮生长因子受体单克隆抗体（EGFR mAb）后,形成具有靶向功能性的QD800-EGFR mAb荧光探针;（3）将QD800-EGFR mAb经金黄地鼠各期癌变模型尾静脉注射入血液循环系统以靶向适配原理显示其颊黏膜动态癌变过程荧光图像。结果 （1）QD800-EGFR mAb能够通过血液循环系统与金黄地鼠颊黏膜癌变细胞上的EGFR特异性的靶向适配并荧光显影;（2）QD800-EGFR mAb显示的荧光图像随着金黄地鼠颊黏膜癌变程度的加深而增强,且能准确地显示癌灶的形状及浸润深度。结论 QD800-EGFR mAb通过与口腔癌不同癌变阶段癌细胞表面的EGFR靶向性适配结合来显示个体化荧光图像,可能对以后临床研究口腔癌的发生发展及转移规律发挥巨大的影响。     

基因打靶与体细胞克隆技术制备乳腺生物反应器的研究进展

在动物乳腺生物反应器研究中,传统方法不可避免地造成基因表达调控元件的人工拼接和外源基因在动物基因组中随机整合所带来的“位置效应”,致使转基因动物外源基因的表达水平不高且差异较大。而基因打靶在外源基因定点整合、消除位点效应方面具有明显优势。体细胞核移植技术可以提前在细胞水平对转基因动物进行筛选,不但可以节省时间,更降低了制备乳腺生物反应器的成本。作者简述了基因打靶与体细胞核移植技术结合制备乳腺生物反应器的优越性、存在问题、解决办法及其应用前景和展望。     

一种用于水稻点突变检测的PCR引物设计方法

介绍了一种用于水稻点突变检测的PCR引物设计方法。利用该方法根据12个候选基因序列设计了12对引物,其中10对引物可以正常工作,能从基因组中扩增出特异性条带,合格率为83%,并且成功地在突变体中检测到点突变。这一方法不仅可以用于水稻,也可以用于其他动植物。同时,对引物设计过程中常见问题进行了讨论并给出了解决办法。     

Protective effect of Yiqi-Wenyang-Huoxue-Huatan formula on HHPH rat brain by suppressing excessive endoplasmic reticulum stress

AIM: To investigate whether Yiqi-Wenyang-Huoxue-Huatan formula (YWHHF) attenuats brain injury induced by hypoxia-hypercapnia pulmonary hypertension (HHPH) in the rats by inhibiting excessive endoplasmic reticulum stress response. METHODS: Healthy SPF male SD rats (n=50) were randomly divided into 5 groups: control group, hypoxia-hypercapnia group, low-dose YWHHF group, middle-dose YWHHF group and high-dose YWHHF group. The rats in control group lived in normal environment, while the rats in other 4 groups were raised for 4 weeks in oxygen tank with low oxygen concentration and high CO₂ concentration. YWHHF was perfused in the rats of low-, middle-and high-dose groups at 0.15, 0.3 and 0.6 g/kg daily, respectively. The rats in hypoxia-hypercapnia group were given isometric distilled water. The surgery was performed on the rats after 4 weeks, and the brain and lung tissues were quickly collected to detect brain water content and observe the morphological changes after mean pulmonary artery pressure recording and heart perfusion. The caspase-3 activity and the apoptotic index of the brain cells were determined. The expression of c-Jun N-terminal kinase (JNK), caspase-12, C/EBP homologous protein (CHOP) and glucose-regulated protein 78 (GRP78) at protein and mRNA levels in brain tissues was detected by Western blot and RT-PCR. RESULTS: Compared with control group, mean pulmonary artery pressure, brain water content, brain apoptotic index, caspase-3 activity, and the protein and mRNA levels of JNK, caspase-12, CHOP and GRP78 in the rest 4 groups were increased, and the brain and lung tissues had obvious damage under light microscope. Compared with hypoxia-hypercapnia group, mean pulmonary artery pressure, brain water content, brain apoptotic index, caspase-3 activity, and the protein and mRNA expression of JNK, caspase-12, CHOP and GRP78 in low-, middle-and high-dose YWHHF groups were decreased, and the pathological damage of the brain and lung tissues was obviously reduced under light microscope. These changes in middle-dose YWHHF group were the most significant. CONCLUSION: YWHHF effectively relieves the brain injury induced by HHPH in rats, which may be associated with inhibiting excessive endoplasmic reticulum stress response.     

簇毛麦3V染色体短臂特异分子标记的开发与应用

普通小麦野生近缘物种簇毛麦(Haynaldia villosa L. 2n=14, VV) 3V染色体短臂（3VS）携带抗小麦条锈病基因。开发高密度的3VS特异标记是准确鉴定和追踪3VS、推进抗病基因转移和利用的重要基础。为了开发更多的均匀分布于3VS染色体臂不同区段的分子标记，本研究利用簇毛麦和小麦品种中国春参考基因组序列与基因注释信息，通过比较3VS与3AS、3BS、3DS同源基因序列内含子序列差异，选择同一内含子长度差异大于10%的区域开发了104个跨越内含子(intron targeting, IT)的分子标记，在簇毛麦、簇毛麦-硬粒小麦双二倍体、硬粒小麦中引1286、中国春、普通小麦-簇毛麦二体异附加系DA1V-DA7V、小麦-簇毛麦T3VS·3DL易位系中进行扩增，筛选出3VS特异、扩增条带清晰且稳定性好的IT分子标记43个。为了验证上述标记的有效性，选择位于3VS不同区段的12个标记对［DS3V(3D)×中国春ph1b］的F₃群体中的148个ph1b纯合的单株进行分析，共筛选得到6种类型涉及3VS的结构变异体。进一步利用GISH/FISH技术对上述材料进行鉴定，结果与分子标记结果相吻合，证明利用外源染色体特异分子标记可有效筛选涉及外源染色体结构变异体。     

脑珊瑚白化率与海水微量元素的相关性分析——美属维尔京群岛珊瑚科学考察（英文）

脑珊瑚(Diploria strigosa)主要分布在大西洋的加勒比海地区的美属维尔京群岛圣约翰岛Great Lameshur Bay,种类很少,珊瑚白化现象严重。该文采用样带场地调查脑珊瑚数量和白化率统计法,结合电感耦合等离子体质谱仪测试技术对海水中微量元素检测分析,对脑珊瑚白化率与微量元素的相关性进行分析,结果表明:水温27.83~28.88℃、p H值8.5~8.7、盐度32.74%~34.04%,海水深度0~4m,微量元素有V、Cr、Rb、Sr、Ag。钒(V)含量低于99.1 ng/ml,脑珊瑚白化率与钒(V)元素含量有弱相关性;铬(Cr)含量低于23.5 ng/ml,脑珊瑚白化率与铬(Cr)元素含量有相关性;银(Ag)含量在34.1~56.3 ng/ml范围,脑珊瑚白化率与银(Ag)元素含量具有很强的相关性。     

Distribution of the Shadoo protein in the ovine brain assessed by immunohistochemistry

Shadow of prion protein is a gene potentially involved in the pathogenesis of prion diseases. However, the Shadoo protein encoded by this gene has not yet been studied in sheep, an important species in prion matters. Therefore, we developed a polyclonal antibody against ovine Shadoo and assessed the presence and distribution of this protein in the ovine brain by immunohistochemistry. The strongest staining level was found in the cerebellum (especially in the Purkinje cells) and in the pons, but cerebrum, hippocampus, pituitary gland, medulla oblongata, thalamus and hypothalamus were also immunopositive. Remarkably, a typical granular pattern was seen in most of the tested brain tissues, which might indicate that Shadoo is primarily expressed at synapses. The results of this study and the availability of an ovine anti-Shadoo antibody can contribute to future research on the function of Shadoo and on its potential involvement in prion diseases.