P-glycoprotein,multidrug resistance and tumor progression

P-glycoprotein (Pgp) is a plasma membrane protein that was first characterised in multidrug resistant cell lines. The occurrence of Pgp in clinical tumors has been widely studied. Recent investigations have begun to focus on the relationship between Pgp detection in tumors and treatment outcome. In several types of tumors, detection of Pgp correlates with poor response to chemotherapy and shorter survival. P-glycoprotein overexpression often occurs upon relapse from chemotherapy but may also occur at the time of diagnosis. Studies of experimental rat liver carcinogenesis have shown that Pgp expression increases in late stages of carcinogenesis, suggesting that Pgp may be involved in tumor progression. While some of the Pgp isoforms are known to transport hydrophobic chemotherapeutic drugs out of tumor cells, the biologic effects of Pgp overexpression in tumor cells are not fully understood, because the spectrum of substrates for Pgp-mediated transport has not been determined. In the rat liver carcinoma model, strong expression of Pgp is associated with a highly vascular stroma, suggesting that Pgp in tumor cells may affect the connective tissue stroma. The regulation of Pgp appears to be complex, and little is known about how it is up-regulated during carcinogenesis. Further studies of the role of Pgp in malignancy may contribute to our understanding of molecular mechanisms which underlie tumor progression.     

Global gene profiling reveals a downregulation of BMP gene expression in experimental atrophic nonunions compared to standard healing fractures.

Nonunion is a challenging problem that may occur following certain bone fractures. However, there has been little investigation of the molecular basis of nonunions. Bone morphogenetic proteins (BMPs) play a significant role in osteogenesis. However, little is known about the expression patterns of BMPs in abnormal bone healing that results in nonunion formation. These facts prompted us to investigate and compare the gene expression patterns of BMPs and their antagonists in standard healing fractures and nonunions using rat experimental models. Standard closed healing fractures and experimental atrophic nonunions produced by periosteal cauterization at the fracture site were created in rat femurs. At postfracture days 3, 7, 10, 14, 21, and 28, total RNA was extracted from the callus of standard healing fracture and fibrous tissue of nonunion (n=4 per each time point and each group). Gene expression of BMPs, BMP antagonists, and other regulatory molecules were studied by methods including Genechip microarray and real-time quantitative RT-PCR. Gene expression of BMP-2, 3, 3B, 4, 6, 7, GDF-5, 7, and BMP antagonists noggin, drm, screlostin, and BAMBI were significantly lower in nonunions compared to standard healing fractures at several time points. Downregulation in expression of osteogenic BMPs may account for the nonunions of fracture. The balance between BMPs and their endogenous antagonists is critical for optimal fracture healing.     

MDR1特异性核酶逆转肝癌多药耐药的实验研究

目的探讨MDR1核酶（N2A＋tRNAi^met-iMDRl-sRz，sRz）在裸鼠体内逆转人肝癌组织多药耐药（MDR）的可行性。方法将原发性MDR人肝癌组织裸鼠原位移植模型第2代随机分为A组（空白对照组：生理盐水40μl＋Lipofect AMINE^TM2000 10μ1）、B组（阴性对照组：N2A＋tRNAi^met 10μg／40μl＋Lipofect AMINE^TM2000 10μl）和C组（核酶组：sRz 10μg／40μl＋LipofectAMINE^TM2000 10μl），均开腹瘤内注射。瘤内注药1周后用表阿霉素15mg／kg腹腔注射，每周1次，连续4周。彩色B超测量肿瘤体积。化疗结束后1周处死裸鼠，RT-PCR、Western blot法检测肿瘤中MDR1 mRNA及其蛋白P-gp的表达。结果C组每次化疗后肿瘤体积均较前缩小（F=659．99，P〈0．05）。除第1次化疗外，其余各次化疗后C组的抑制率均高于A、B组（F=35．36，12．77，97．60，P〈0．05）。化疗结束后，C组与瘤源以及A、B组相比，肿瘤组织中MDR1 mRNA和P-gp的表达明显降低（F=45．36，3590．40，P〈0．05）。结论sRz可有效降低肝癌细胞表达MDR1 mRNA和P-gp，一定程度上逆转MDR，提高E-ADM的化疗效果。单纯E-ADM化疗可使肝癌组织MDR1 mRNA和P-gp表达升高，诱导MDR的产生。     

人钠/二羧酸协同转运蛋白3基因调控肾小管上皮细胞线粒体膜电位的研究

目的 观察人钠／二羧酸协同转运蛋白3(hNaDC3，)对人肾脏近曲小管上皮细胞(HKC)线粒体膜电位的变化及其对细胞能量代谢的影响。方法 应用亚克隆技术构建正义pcDNA3-hNaDC3和反义pcDNA3-AhNaDC3两个真核表达载体，通过脂质体LipofectAMINE将pcDNA3-hNaDC3及pcDNA3-AhNaDC3转染至HKC细胞。克隆筛选后，用RT—PCR、Northern印迹及Western印迹鉴定外源基因的整合和表达。荧光探针JC-1观察各细胞系线粒体膜电位的变化。结果 外源hNaDC3基因稳定整合到HKC细胞基因组中，并获得高、低表达。转染正义hNaDC3cDNA的HKC细胞线粒体膜电位降低，JC-1在线粒体内形成单体，发出绿色荧光；而转染反义hNaDC3cDNA的HKC细胞线粒体膜电位略微升高，JC-1形成聚合体，发出红色荧光。结论 hNaDC3过表达引起线粒体膜电位降低，反义hNaDC3则使线粒体膜电位略微升高。提示NaDC3可能通过使线粒体膜电位下降，参与了细胞能量代谢。     

血清蛋白变化对鼻咽癌患者预后判断的意义

本文研究了155例鼻咽癌血清蛋白电泳α_2/β的比值变化。结果表明,正常组(α_2/β<1)5年生存率明显高于异常组(α_2/β≥1)。尽管两组在治疗结束时疗效相似(完全缓解率为66.1％和65.2％),但5年内复发率前者为30.6％.后者为80.0％。因此,我们认为血清蛋白电泳不同于临床分期或其他预后因素,对判断鼻咽癌病人的预后方面更有价值。     

地塞米松对哮喘豚鼠肺组织Giα、Gqα的抑制作用

目的　研究糖皮质激素对豚鼠哮喘模型肺组织G蛋白α亚族表达的影响。方法　 18只豚鼠随机分为正常对照组(NS) ,哮喘组 (AS)和地塞米松治疗组 (DEX)。AS组用卵蛋白皮下及腹腔注射致敏 ,卵蛋白重复雾化吸入激发复制豚鼠哮喘模型。DEX组在每次激发前腹腔内注射地塞米松 2mg/kg进行干预。用Westernblot分析法测定豚鼠肺组织G蛋白α亚族的表达水平。结果　三组豚鼠肺组织Giα水平分别为NS组 10 0 % ,AS组 (15 3± 18) % ,DEX组 (113± 18) % ,AS组与NS组比较、DEX组与AS组比较差别显著 (P <0 .0 5 )。三组豚鼠肺组织Gqα水平分别为NS组 10 0 % ,AS组 (15 1± 19) % ,DEX组 (98± 4 ) % ,AS组与NS组比较、DEX组与AS组比较差别显著 (P <0 .0 5 )。在同样的条件下Gsα水平无显著变化 (P >0 .0 5 )。地塞米松能够显著抑制哮喘豚鼠肺组织中Giα、Gqα的高表达。结论　肺组织Giα ,Gqα的高表达变化参与了豚鼠哮喘模型的病理性信号传导 ,抑制哮喘肺组织Giα、Gqα的高表达可能是糖皮质激素治疗哮喘的机制之一。     

Differential Distribution of MAP1a and Aldolase c in Adult Mouse Cerebellum

MAP1a is a microtubule-associated protein with an apparent molecular weight of 360 kDa that is found in the axonal and dendritic processes of neurons. Two monoclonal anti-MAP1a antibodies, anti-A and anti-BW6, revealed different epitope distributions in the adult mouse cerebellum. Anti-A stained Purkinje and granule cells uniformly throughout the cerebellum. In contrast, anti-BW6 selectively stained the dendrites of a subset of Purkinje cells, revealing parasagittal bands of immunoreactivity in the molecular layer. The compartmentation of the BW6 epitope was compared to the Purkinje cells as revealed by immunostaining with anti-zebrin II, a well known antigen expressed selectively by bands of Purkinje cells. The anti-BW6 staining pattern was complementary to the zebrin II bands, the zebrin II^- Purkinje cells having BW6⁺ dendrites. These results demonstrate that MAP1a is present in two forms in the mouse cerebellum, one of which is segregated into parasagittal bands. This may indicate a unique MAP1a isoform or may reflect differences in the metabolic states of Purkinje cell classes, and regional differences in their functions.     

核仁形成区嗜银蛋白定量对结肠癌分型、分级的诊断意义

对１１４例不同类型和级别的结肠癌进行银染核仁形成区嗜银蛋白（Ａｇ－ＮＯＲｓ）形态定量研究．结果表明，Ａｇ－ＮＯＲｓ数量、大小和分布在结肠未分化癌和印戒细胞癌组与管状腺癌和粘液腺癌组比较，有明显差异．管状腺癌随分化程度的降低，Ａｇ－ＮＯＲｓ的数量、形态、大小和分布也发生等级性变化．结果提示，Ａｇ－ＮＯＲｓ形态定量的数量、形态、大小和分布４项指标对结肠癌分型有一定意义，对管状腺癌分级有较好的诊断价值。     

急进高原时尿中微量蛋白的变化

本文观察了从西宁地区(2260m)快速进入高海拔地区(3700m(5天)和4600m(10天)时,尿中微量蛋白的排泄变化,结果表明,在西宁地区尿中微量蛋白水平与平原无异,到达3700m时,尿中AIb排出明显增多(P<0.01),至4600m时AIb排出更多(与3700m相比P<0.001),IgM也有增多趋势,而世居藏族除AIb明显增多外,尿中IgM亦较西宁地区明显增高(P<0.05),作者以为尿中微量蛋白的排泄变化有高度和时间界限,发生机理可能与低氧环境所致的交感、肾上腺内分泌系统的应激反应以及血液流变学改变有关。     

Cytochemical characteristics of neurons in the trigeminal mesencephalic nucleus of hatchling chicks

The goal of the present study was to identify cytochemical markers characteristic of muscle afferents in hatchling chicks. To this end, we stained neurons in the trigeminal mesencephalic nucleus with a variety of markers that label subsets of neurons in avian dorsal root ganglia. We found that trigeminal mesencephalic neurons are surprisingly heterogeneous in their cytochemical make-up, expressing, to varying degrees, substance P, cholecystokinin, carbonic anhydrase, calbindin D-28k, parvalbumin, and S-100β. Calbindin D28k and S-100β appeared to be expressed equally in medial and lateral divisions of the trigeminal mesencephalic nucleus. In contrast, substance P- and cholecystokinin-immunoreactive neurons were more abundant in the medial division, whereas carbonic anhydrase activity and parvalbumin immunoreactivity were stronger in the lateral division. We were unable to detect met-enkephalin, neuropeptide Y, calcitonin gene-related peptide, vasoactive intestinal peptide, somatostatin, γ-aminobutyric acid, or tyrosine hydroxylase in the trigeminal mesencephalic nucleus. Moreover, these neurons did not appear to bind the lectin Dolichos biflorus agglutinin. The heterogeneity of expression of markers among trigeminal mesencephalic nucleus neurons, especially between neurons in the medial and lateral divisions, suggests that these neurons are functionally diverse.