Expression of PDE11A in normal and malignant human tissues.

Cyclic nucleotide phosphodiesterase 11A (PDE11A) is the newest member in the PDE family. Although the tissue distribution of PDE11A mRNA has been shown, its protein expression pattern has not been well studied. The goal of this report is to investigate the distribution of PDE11A proteins in a wide range of normal and malignant human tissues. We utilized a polyclonal antibody that recognized all four PDE11A isoforms. Its specificity was demonstrated by Western blot analysis on a recombinant human PDE11A protein and native PDE11A proteins in various human tissues. Immunohistochemistry showed that PDE11A is widely expressed. Various degrees of immunoreactivity were observed in the epithelial cells, endothelial cells, and smooth muscle cells of all tissues examined. The highest expression was in the epithelial, endothelial, and smooth muscle cells of the prostate, Leydig, and spermatogenic cells of the testis, the tubule epithelial cells in the kidney, the epithelial and endothelial cells in the adrenal, the epithelial cells and macrophages in the colon, and the epidermis in the skin. Furthermore, PDE11A expression was also detected in several human carcinomas. Our results suggest that PDE11A might be involved in multiple physiological processes in various organs via its ability to modulate intracellular cAMP and cGMP levels.     

Plasma glycoprotein profiling for colorectal cancer biomarker identification by lectin glycoarray and lectin blot

Colorectal cancer (CRC) remains a major worldwide cause of cancer-related morbidity and mortality largely due to the insidious onset of the disease. The current clinical procedures utilized for disease diagnosis are invasive, unpleasant, and inconvenient; hence, the need for simple blood tests that could be used for the early detection of CRC. In this work, we have developed methods for glycoproteomics analysis to identify plasma markers with utility to assist in the detection of colorectal cancer (CRC). Following immunodepletion of the most abundant plasma proteins, the plasma N -linked glycoproteins were enriched using lectin affinity chromatography and subsequently further separated by nonporous silica reversed-phase (NPS-RP)-HPLC. Individual RP-HPLC fractions were printed on nitrocellulose coated slides which were then probed with lectins to determine glycan patterns in plasma samples from 9 normal, 5 adenoma, and 6 colorectal cancer patients. Statistical tools, including principal component analysis, hierarchical clustering, and Z-statistics analysis, were employed to identify distinctive glycosylation patterns. Patients diagnosed with colorectal cancer or adenomas were shown to have dramatically higher levels of sialylation and fucosylation as compared to normal controls. Plasma glycoproteins with aberrant glycosylation were identified by nano-LC-MS/MS, while a lectin blotting methodology was used to validate proteins with significantly altered glycosylation as a function of cancer progression. The potential markers identified in this study for diagnosis to distinguish colorectal cancer from adenoma and normal include elevated sialylation and fucosylation in complement C3, histidine-rich glycoprotein, and kininogen-1. These potential markers of colorectal cancer were subsequently validated by lectin blotting in an independent set of plasma samples obtained from 10 CRC patients, 10 patients with adenomas, and 10 normal subjects. These results demonstrate the utility of this strategy for the identification of N -linked glycan patterns as potential markers of CRC in human plasma, and may have the utility to distinguish different disease states.     

Parvovirus infection-induced cell death and cell cycle arrest

The cytopathic effects induced during parvovirus infection have been widely documented. Parvovirus infection-induced cell death is often directly associated with disease outcomes (e.g., anemia resulting from loss of erythroid progenitors during parvovirus B19 infection). Apoptosis is the major form of cell death induced by parvovirus infection. However, nonapoptotic cell death, namely necrosis, has also been reported during infection of the minute virus of mice, parvovirus H-1 and bovine parvovirus. Recent studies have revealed multiple mechanisms underlying the cell death during parvovirus infection. These mechanisms vary in different parvoviruses, although the large nonstructural protein (NS)1 and the small NS proteins (e.g., the 11 kDa of parvovirus B19), as well as replication of the viral genome, are responsible for causing infection-induced cell death. Cell cycle arrest is also common, and contributes to the cytopathic effects induced during parvovirus infection. While viral NS proteins have been indicated to induce cell cycle arrest, increasing evidence suggests that a cellular DNA damage response triggered by an invading single-stranded parvoviral genome is the major inducer of cell cycle arrest in parvovirus-infected cells. Apparently, in response to infection, cell death and cell cycle arrest of parvovirus-infected cells are beneficial to the viral cell lifecycle (e.g., viral DNA replication and virus egress). In this article, we will discuss recent advances in the understanding of the mechanisms underlying parvovirus infection-induced cell death and cell cycle arrest.     

Overexpression of fucosyltransferase IV in A431 cell line increases cell proliferation

Fucosyltransferase IV is an essential enzyme that catalyzes the synthesis of fucosylated oligosaccharides by transferring GDP-fucose to the terminal N-acetylglucosamine with the alpha1,3-linkage. Lewis Y oligosaccharide has a terminal alpha1,3-linked fucose residue and elevation of Lewis Y level is seen in many epithelial cancers. The mechanism of Lewis Y elevation in neoplastic cells is still largely unknown. To study the impact of fucosyltransferase IV on Lewis Y expression and its role on neoplastic cell proliferation, a pEGFP-N1-FUT4 recombinant plasmid was developed and stably transfected into A431 cells. We found that fucosyltransferase IV overexpression promoted cell proliferation and increased the expression of proliferating cell nuclear antigen that correlated with Lewis Y augmentation. Cell cycle analysis demonstrated that fucosyltransferase IV overexpression facilitated cell cycle progression. In conclusion, fucosyltransferase IV overexpression augments Lewis Y expression to trigger neoplastic cell proliferation. These studies suggest that fucosyltransferase IV may serve as a potential therapeutic target for the treatment of Lewis Y-positive epithelial cancers.     

人X染色体间期细胞遗传学研究

应用人X染色体α卫星DNA探针进行X染色体正常或异常个体的外周血淋巴细胞染色体和间期核的原位杂交,在R显带的中期分裂相上,绝大部分杂交颂粒位于X染色体着丝粒区(p11→q11);在间期核内则显现与X染色体数相一致的银颗粒簇,其中相当部分位于核边缘区。实验结果表明,用原位杂交来检测X染色体数目,比记数Barr小体的方法可靠。本文还就α卫星DNA探针在间期细胞遗传学方面广泛的应用做了讨论。     

猪产仔数分子标记及其效应分析

初步确定了2个新的猪产仔数分子标记,雌激素受体基因ESR的第8外显子处的ESRB位点、催乳素受体基因的第7外显子的FSHRB位点。通过比较和分析多个产仔数的效应,初步确定了4个有利于产仔数提高的分子标记基因型,基因位点ESR、FSHRB的基因型BB的产仔数显著地高于AB、AA型;位点ESRB、PRLR的基因型AA的产仔数显著地高于AB、BB型;4个基因位点多态性与仔猪生长性能、母猪乳头数不存在显著的影响。     

基于免疫纳米颗粒强化的压电免疫传感器检测大肠杆菌O157︰H7

摘要：【目的】结合纳米技术建立检测大肠杆菌（Escherichia coli）O157︰H7高灵敏检测技术。【方法】采用化学共沉淀法制备出核心粒径约为10 nm的免疫纳米磁颗粒,柠檬酸钠还原法制备粒径约为20 nm的免疫胶体金。压电免疫传感器通过金黄色葡萄球菌蛋白A（Protein A from Staphylococcus aureus SPA）法将抗体固定于石英晶振上,两种免疫纳米颗粒借助不同的抗体连接于传感器上对检测频率信号进行放大。【结果】SPA在石英晶振上的最佳固定浓度和时间为1.2 mg/mL和40 min,抗体的最佳固定浓度和时间为1.0 mg/mL和60 min。压电免疫传感器通过两种免疫纳米颗粒的放大作用,使其对大肠杆菌O157︰H7的检测限从104 cfu/mL提高到101 cfu/mL。【结论】免疫纳米颗粒强化对压电免疫传感器的检测频率信号具有很好的放大效应,可以明显提高其检测灵敏度。     

禄丰古猿地点的小哺乳动物化石

过去十年,在禄丰古猿地点作了多次的发掘和筛洗,采集到包括食虫类,翼手类,啮齿类和兔形类在内的一千余件标本,使这一小哺乳动物的化石组合从1979年首次报道的6种增加到了38种,成为我国南方新第三纪中期最丰富而有代表性的小哺乳动物群。材料的初步鉴定表明:动物群中有代表我国首次发现的化石科和属;并有相当数量与华北、印巴次大陆和欧洲有密切亲缘关系的类型;其地质时代属最晚中新世保德期,相当于欧洲陆相哺乳动物分期的土洛里期(Turolian)。     

2014生物质炼制专刊序言

生物质是自然界最丰富的含碳有机大分子功能体,它有望通过"生物炼制"实现"石油炼制"的辉煌。但是由于生物质资源本身及其转化过程的复杂性,生物质产业虽备受关注,却被认为是遥远的未来产业。传统的生物质资源化利用思路都是先耗费一定的能量破坏生物质结构,然后再进行转化,不仅没有考虑到产品的功能需求,而且过程的原子经济性不高。如何实现化学键更加复杂的固相木质纤维素生物质炼制是实现生物质产业的关键和难点。理想的生物质炼制的目的是以最大得率分离木质纤维原料中各个组分,以尽可能地保持分子的完整性,最大可能地优化利用和最终实现最大价值。这就要求生物质炼制应当是基于原料结构、过程转化和产品特点三者的关联,面向原料、面向过程、面向产品的炼制过程。本期专刊报道了我国生物质炼制技术领域专家学者在原料炼制、炼制技术、组分转化等领域取得的最新研究进展。     

毕赤酵母发酵生产颗粒性乙肝表面抗原的条件优化

乙型肝炎病毒的流行对人们的生命健康造成了极大的威胁, 而有效准确的诊断和预防性疫苗是阻止其流行的主要手段, 乙肝表面抗原是诊断试剂和疫苗的主要成分。本试验在构建稳定表达HBsAg的毕赤酵母菌株后, 对其发酵条件进行了研究。采用摇瓶分批培养方法, 探讨了不同培养基、溶解氧、诱导物甲醇的浓度以及pH值等因素对菌体生长与重组蛋白表达的影响。在10 L发酵罐上采用分批补料培养的方法研究了进行扩大培养生产重组HBsAg。结果表明, FBS无机盐合成培养基是理想的工业发酵培养基, 溶解氧对菌体的生长与表达有显著的影响, 甲醇诱导最佳终浓度为1% (V/V), 发酵的最适pH值为5.4~6.0。发酵罐放大培养后, ELISA和 SDS-PAGE分析表明重组HBsAg获得了高效表达, 最终菌体生物量达到310 OD600, 表达量达到27 mg/L。电子显微镜观察表达重组乙肝抗原可以自组装为22 nm类病毒颗粒, 为HBV的新一代早期血清学诊断和疫苗的大规模生产提供了一定的参考。