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151.
152.
R. T. Hayden J. R. Uhl X. Qian M. K. Hopkins M. C. Aubry A. H. Limper R. V. Lloyd F. R. Cockerill 《Journal of clinical microbiology》2001,39(7):2618-2626
We developed a rapid thermocycling, real-time detection (also known as real-time PCR) method for the detection of Legionella species directly from clinical specimens. This method uses the LightCycler (Roche Molecular Biochemicals, Indianapolis, Ind.) and requires approximately 1 to 2 h to perform. Both a Legionella genus PCR assay and Legionella pneumophila species-specific PCR assay were designed. A total of 43 archived specimens from 35 patients were evaluated, including 19 bronchoalveolar lavage (BAL) specimens and 24 formalin-fixed, paraffin-embedded open lung biopsy specimens. Twenty-five of the specimens were culture-positive for Legionella (9 BAL specimens and 16 tissue specimens). BAL specimens were tested by LightCycler PCR (LC-PCR) methods and by a direct fluorescent antibody (DFA) assay, which detects L. pneumophila serogroups 1 to 6 and several other Legionella species. Tissue sections were tested by the two LC-PCR methods, by DFA, by an in situ hybridization (ISH) assay, specifically designed to detect L. pneumophila, and by Warthin-Starry (WS) staining. The results were compared to the "gold standard" method of bacterial culture. With BAL specimens the following assays yielded the indicated sensitivities and specificities, respectively: Legionella genus detection by Legionella genus LC-PCR, 100 and 100%; Legionella genus detection by DFA assay, 33 and 100%; and L. pneumophila detection by L. pneumophila species-specific LC-PCR, 100 and 100%. With open lung biopsy specimens the following assays yielded the indicated sensitivities and specificities, respectively: Legionella genus detection by LC-PCR 68.8 and 100%; Legionella genus detection by DFA assay, 44 and 100%; Legionella genus detection by WS staining, 63 and 100%; L. pneumophila species-specific detection by LC-PCR, 17 and 100%; and L. pneumophila species-specific detection by ISH, 100 and 100%. The analytical sensitivity of both LC-PCR assays was <10 CFU/reaction. LC-PCR is a reliable method for the direct detection of Legionella species from BAL specimens. The Legionella genus LC-PCR assay could be performed initially; if positive, L. pneumophila species-specific LC-PCR could then be performed (if species differentiation is desired). The speed with which the LC-PCR procedure can be performed offers significant advantages over both culture-based methods and conventional PCR techniques. In contrast, for the methods evaluated, culture was the best for detecting multiple Legionella species in lung tissue. WS staining, Legionella genus LC-PCR, and L. pneumophila species-specific ISH were useful as rapid tests with lung tissue. 相似文献
153.
154.
利用木瓜蛋白酶分别制备了抗汉坦病毒鼠源性单克隆抗体(mAb)3G1和3D8F(ab′)2片段。通过WaterDEAE40HR阴离子交换色谱柱纯化后,非还原型SDSPAGE呈现单一条带,相对分子质量(Mr)约100000。两种F(ab′)2片段均具有良好的血凝抑制活性、病毒中和活性及对汉坦病毒感染乳鼠的保护作用 相似文献
155.
一种可生物降解温度敏感型聚乙二醇-聚己内酯-聚乙二醇水凝胶的合成和表征 总被引:1,自引:0,他引:1
合成了一系列分子量较低的聚乙二醇.聚己内酯-聚乙二醇(Poly(ethylene glycol)-Polycaprolactone-Poly(ethylene glycol),PEG-PCL—PEG)三嵌段共聚物。分别采用FTIR和1H—NMR对其结构进行了表征。所合成的PEG-PCL-PEG共聚物具有良好的水溶性,当水溶液浓度高于临界凝胶浓度(Critical gel concentration,CGC)时,随着温度的变化聚合物水溶液会呈现特有的凝胶-溶胶转变。研究了共聚物亲水疏水链段的比例和长度,以及热历史等对凝胶-溶胶转变行为的影响。通过调节上述条件,可以在一定程度上拓宽凝胶-溶胶转变温度范围,有助于PEG—PCL-PEG水凝胶在可注射药物控制释放系统等方面的应用。 相似文献
156.
SH2A 基因对细胞信号转导的影响及其亚细胞定位 总被引:3,自引:0,他引:3
目的 研究Src同源域2(src homology 2,SH2)A基因在细胞信号转导中的作用并进行亚细胞定位。方法 通过RT—PCR方法扩增SH2A cDNA编码序列,构建真核重组表达载体pcDNA3.1-SH2A,利用脂质体转染肝癌Bel7402细胞、COS7细胞,检测蛋白酶C(protein kinaseC,PKC)、酪氨酸蛋白激酶(tyrosine protein kinase,TPK)、丝裂原激活蛋白激酶(mitogen activated protein kinase,MAPK)活性的改变;另构建pEGEP—SH2A,转染同前,荧光显微镜观察荧光定位。结果 测序结果显示真核重组表达载体pcDNA3.1-SH2A及pEGFP—SH2A中均含有SH2AcDNA编码序列;肝癌Bel7402细胞、COS7细胞转染pcDNA3.1-SH2A后,胞浆PKC的活性下降了40%左右,MAPK和TPK活性未见明显改变。荧光显微镜观察发现SH2A基因在细胞质中表达。结论 SH2A基因编码蛋白在PKC信号转导通路中起抑制作用;SH2A基因编码蛋白定位于细胞质。 相似文献
157.
Qian Wei Donatella Chionna Mariano Pracella 《Macromolecular chemistry and physics.》2005,206(7):777-786
Summary: Blends of polyamide‐6 (PA6) and low density polyethylene (LDPE) were compatibilized by melt mixing with various polyolefins functionalized with glycidyl methacrylate (GMA), i.e., GMA grafted LDPE (LDPE‐g‐GMA), GMA grafted styrene‐ethylene/butylene‐styrene block copolymer (SEBS‐g‐GMA) and ethylene‐co‐glycidyl methacrylate copolymer (E‐GMA). Blends with PA6/LDPE composition ratios of 25/75 and 75/25 wt.‐%/wt.‐% were prepared in a Brabender internal mixer and their properties were evaluated by SEM, rheological measurements and DSC. Morphological investigation by SEM showed a neat improvement of phase dispersion and interfacial adhesion in all compatibilized blends when compared to PA6/LDPE binary blends. The variation of the dispersed phase size was analyzed as a function of blend composition, compatibilizer concentration and GMA content. The emulsification curves of compatibilized blends showed that the equilibrium size of dispersed particles at the saturation concentration of copolymer was lower when PA6 was the major component. The finest dispersion of the LDPE phase (<0.25 μm) was observed in the presence of SEBS‐g‐GMA copolymer. LDPE‐g‐GMA and E‐GMA displayed a similar compatibilizing efficiency. In all cases, the blends with a polyamide matrix presented a marked rise in torque and melt viscosity with increasing compatibilizer content. These effects were accounted for by a reaction between the epoxide groups of LDPE‐g‐GMA and the carboxyl/amine end‐groups of PA6, leading to the formation of an interchain graft copolymer. The phase transition processes of PA6 in the blends were influenced by the compatibilizer content and the interfacial interactions between the polymer components, suggesting a different role for the compatibilizer at the PA6/LDPE interface.
158.
Pituitary Adenylate-Cyclase-Activating Polypeptide (PACAP) Binding Sites and PACAP/Vasoactive Intestinal Polypeptide Receptor Expression in Human Pituitary Adenomas 下载免费PDF全文
159.
Xin-Tong Wei Gui-Juan Feng Hong Zhang Qian Xu Jing-Jing Ni Min Zhao Xiao-Lin Yang Qing Tian Hui Shen Rong Hai Hong-Wen Deng Lei Zhang Yu-Fang Pei 《European journal of human genetics : EJHG》2021,29(4):553
Osteoporosis and obesity are two severe complex diseases threatening public health worldwide. Both diseases are under strong genetic determinants as well as genetically correlated. Aiming to identify pleiotropic genes underlying obesity and osteoporosis, we performed a bivariate genome-wide association (GWA) meta-analysis of hip bone mineral density (BMD) and total body fat mass (TBFM) in 12,981 participants from seven samples, and followed by in silico replication in the UK biobank (UKB) cohort sample (N = 217,822). Combining the results from discovery meta-analysis and replication sample, we identified one novel locus, 17q21.31 (lead SNP rs12150327, :g.44956910G > A, discovery bivariate P = 4.83 × 10−9, replication P = 5.75 × 10−5) at the genome-wide significance level (ɑ = 5.0 × 10−8), which may have pleiotropic effects to both hip BMD and TBFM. Functional annotations highlighted several candidate genes, including KIF18B, C1QL1, and PRPF19 that may exert pleiotropic effects to the development of both body mass and bone mass. Our findings can improve our understanding of the etiology of osteoporosis and obesity, as well as shed light on potential new therapies.Subject terms: NC_000017.11Genome-wide association studies, Gene expression profiling 相似文献
160.
Pkd2 haploinsufficiency alters intracellular calcium regulation in vascular smooth muscle cells 总被引:7,自引:0,他引:7
Qian Q Hunter LW Li M Marin-Padilla M Prakash YS Somlo S Harris PC Torres VE Sieck GC 《Human molecular genetics》2003,12(15):1875-1880
Autosomal-dominant polycystic kidney disease is a multiorgan disease and its vascular manifestations are common and life-threatening. Despite this, little is known about their pathogenesis. Somatic mutations to the normal PKD allele in cystic epithelia and cyst development associated with the unstable Pkd2(WS25) allele suggest a two-hit model of cystogenesis. However, it is unclear if this model can account for the cardiovascular pathology or if haploinsufficiency alone is disease-associated. In the present study, we found a decreased polycystin-2 (PC2, protein encoded by Pkd2 gene) expression in Pkd2( +/-) vessels, roughly half the wild-type level, and an enhanced level of intracranial vascular abnormalities in Pkd2 (+/-) mice when induced to develop hypertension. Consistent with these observations, freshly dissociated Pkd2 (+/-) vascular smooth muscle cells have significantly altered intracellular Ca(2+) homeostasis. The resting [Ca(2+)](i) is 17.1% lower in Pkd2 (+/-) compared with wild-type cells (P=0.0003) and the total sarcoplasmic reticulum Ca(2+) store (emptied by caffeine plus thapsigargin) is decreased (P<0.0001). The store operated Ca(2+) (SOC) channel activity is also decreased in Pkd2 (+/-) cells (P=0.008). These results indicate that inactivation of just one Pkd2 allele is sufficient to significantly alter intracellular Ca(2+) homeostasis, and that PC2 is necessary to maintain normal SOC activity and the SR Ca(2+) store in VSMCs. Based on these findings, and the fact that [Ca(2+)](i) signaling is essential to the regulation of contraction, production and secretion of extracellular matrix, cellular proliferation and apoptosis, we propose that the abnormal intracellular Ca(2+) regulation associated with Pkd2 haploinsufficiency is directly related to the vascular phenotype. 相似文献