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111.
Ji-Su Li Shu-Ping Tong Ludmila Vitvitski Christian Trpo 《Journal of medical virology》1995,45(2):151-155
Hepatitis C virus (HCV) exhibits considerable sequence variability and circulates in the blood at extremely low levels. Current methods for detecting HCV RNA are based mostly on nested polymerase chain reaction (PCR), in which part of the first amplification product is reamplified in the second tube by an internal primer pair. A novel nested PCR method was developed in which the two successive amplification processes are carried out in the same tube with a single step of physical manipulation. Careful selection of highly conserved sequences of the 5′ noncoding region as primers enabled successful detection of all three major genotypes circulating in France, including the one with variation in this region. Retaining high sensitivity of the conventional nested PCR, the novel method reduced greatly the risk of carry-over contaminations. It was also cost- and time-saving. The one-step nested PCR method is especially suitable for routine diagnosis of HCV infection in clinical laboratories. © 1995 Wiley-Liss, Inc. 相似文献
112.
Effect of stretch on calcium channel currents recorded from the antral circular myocytes of guinea-pig stomach 总被引:7,自引:0,他引:7
Wen Xie Xu Sung Joon Kim Sang Jeong Kim Insuk So Tong Mook Kang Jong Chul Rhee Ki Whan Kim 《Pflügers Archiv : European journal of physiology》1996,432(2):159-164
The effect of membrane stretch on voltage-activated Ba2+ current (I
Ba) was studied in antral circular myocytes of guinea-pig using the whole- cell patch-clamp technique. The changes in cell volume
were elicited by superfusing the myocytes with anisosmotic solutions. Hyposmotic superfusate (202 mosmol/l) induced cell swelling
and increased peak values of I
Ba at 0 mV (from −406.6 ± 45.5 pA to −547.5 ± 65.6 pA, mean ± SEM, n = 8) and hyperosmotic superfusate (350 mosmol/l) induced cell shrinkage and decreased peak values of I
Ba at 0 mV (to −269.5 ± 39.1 pA, n = 8). Such changes were reversible and the extent of change was dependent on the osmolarity of superfusate. The values of
normalized I
Ba at 0 mV were 1.43 ± 0.04, 1.30 ± 0.06, 1.23 ± 0.04, 1.19 ± 0.04, 1 and 0.68 ± 0.06 at 202, 220, 245, 267, 290 and 350 mosmol/l,
respectively (n = 8). I
Ba was almost completely blocked by nicardipine (5 μM) under hyposmotic conditions. The values of steady-state half-inactivation
voltage (−37.7 ± 3.3 and −36.5 ± 2.6 mV, under control and hyposmotic conditions, respectively) or the half-activation voltage
(−13.6 ± 2.3 and −13.9 ± 1.9 mV) of I
Ba were not significantly changed (P > 0.05, n = 6). Cell membrane capacitance was slightly increased from 50.00 ± 2.86 pF to 50.22 ± 2.82 pF by a hyposmotic superfusate
(P < 0.05, n = 6). It is suggested that cell swelling increases voltage-operated L-type calcium channel current and that such a property
is related to the response of gastric smooth muscle to mechanical stimuli.
Received: 14 November 1995/Received after revision and accepted: 8 January 1996 相似文献
113.
180例慢性粒细胞白血病骨髓染色体畸变及意义 总被引:3,自引:0,他引:3
目的 研究骨髓染色体畸变与慢性粒细胞白血病 (慢粒 )不同病期的相关性及其临床意义。方法 采取新鲜骨髓 ,进行短期培养 (48小时或 12小时预加秋水仙素培养 )制备染色体标本 ,分析 180例慢粒患者慢性期、加速期、急变期骨髓染色体的变异情况。结果 180例慢粒患者 ,其中 16 4例Ph染色体阳性 (Ph ) ,占 91 1% ;16例Ph染色体阴性(Ph - ) ,占 8 9%。慢性期 133例Ph ,占总数的 73 9% ;加速期 9例Ph ,占总数 5 0 % ;急变期 2 2例Ph ,占总数12 2 %。 16 4例Ph 慢粒患者中 2 5例 (15 2 % )伴有额外染色体异常 ,其中慢性期占 4 2 7%、加速期占 2 4 %、急变期占8 5 3%。结果表明随着病情的进展 ,Ph 阳性率明显增高 ,其它染色体异常率也明显增高。结论 骨髓染色体畸变与慢粒白血病的病程进展、疾病预后有着十分重要的相关性 ,随着病情的演变 ,骨髓染色体畸变趋于复杂化。进行染色体分析对临床诊断及治疗慢粒具有十分重要的指导意义。 相似文献
114.
Chan PK To KF Lo AW Cheung JL Chu I Au FW Tong JH Tam JS Sung JJ Ng HK 《Journal of medical virology》2004,74(1):1-7
Severe acute respiratory syndrome coronavirus (SARS-CoV) can produce gastrointestinal symptoms. The intestinal tract is the only extrapulmonary site where viable viruses have been detected. This study examined seven established human intestinal cell lines, DLD-1, HCT-116, HT-29, LoVo, LS-180, SW-480 and SW-620, for their permissiveness to SARS-CoV infection. The results showed that only LoVo cells were permissive to SARS-CoV infection as evident by positive findings from indirect immunofluorescence staining for intracellular viral antigens, in situ hybridization for intracellular viral RNA, and electron microscopy for intracellular viral particles. In contrast to Vero cells, SARS-CoV did not produce cytopathic effects on LoVo cells. However, LoVo cells were found to be highly permissive for productive infection with a high viral titre (>3 x 10(7) viral copies/ml) produced in culture supernatant following a few days of incubation. SARS-CoV established a stable persistent chronic infection that could be maintained after multiple passages. Being a cell line of human origin, LoVo cells could be a useful in vitro model for studying the biology and persistent infection of SARS-CoV. Our results on the expression of angiotensin-converting enzyme 2 (ACE2), a recently identified cellular receptor for SARS-CoV, in these cell lines indicated that it might not be the sole determinant for cells to be susceptible to SARS-CoV infection. 相似文献
115.
骨形成蛋白-7对人肾小管上皮细胞株HK-2凋亡的影响 总被引:1,自引:0,他引:1
研究不同浓度的骨成形蛋白 7(BMP 7)对人近曲小管上皮细胞株HK 2凋亡的影响并初步探讨其可能的机制。应用Hoechst 332 5 8荧光染色、DNA电泳检测凋亡细胞的形态学和生化改变 ,流式细胞仪定量检测细胞的凋亡率 ,并观察凋亡特异性蛋白酶Caspase 3活性变化的情况。结果显示 ,HK 2细胞加入 1 0ng/mlTGF β处理 2 4h后 ,可明显抑制细胞生长并诱导细胞凋亡 ,呈现明显的凋亡形态学改变 ,胞核或核质内可见浓度致密的颗粒状荧光 ,DNA电泳呈现典型的“梯形条带” ,而预先加入 5 0ng/ml、 1 0 0ng/mlBMP 7共处理 2 4h后 ,可减轻TGF β对细胞的抑制作用 ,凋亡细胞亦减少 ,Caspase 3蛋白酶活性明显下降 ,细胞凋亡率下降。提示BMP 7对人肾小管上皮细胞凋亡的具有抑制作用 ,可能是通过抑制Caspase蛋白酶活性而实现的 相似文献
116.
目的测定氧化修饰低密度脂蛋白(oxLDL)在子痫前期中的变化,探讨其意义。方法采用酶联免疫吸附法(ELISA)测定子痫前期重度组、子痫前期轻度组及对照组血清oxLDL水平。结果(1)3组孕妇血清oxLDL比较,见子痫前期重度组升高(49.35±4.22)μg/d l,与对照组(31.71±3.51)μg/d l比较有显著差异(P<0.01);子痫前期轻度组(39.12±2.37)μg/d l升高,与对照组比较有显著差异(P<0.05);子痫前期重度组与轻度组比较无显著差异(P>0.05)。(2)子痫前期重度组血清oxLDL与收缩压、舒张压呈正相关(r=0.51,P<0.01;r=0.47,P<0.05)。结论研究提示oxLDL可能是子痫前期发病的重要因素。 相似文献
117.
118.
Fa-Ten Kao Suhong Tong Yiping Shen Jingwei Yu 《Somatic Cell and Molecular Genetics》1996,22(3):191-199
Three region-specific libraries for the entire human chromosome 18 were constructed using microdissection and MboI linker-adaptor
microcloning techniques. The libraries included 18pter-p11.1 (designated 18P library), 18q 11.1-q12.3 (18Q1 library), and
18q21.1-qter (18Q2 library). Samples of the microclones from each library were analyzed in detail. The insert sizes ranged
between 50–600 bp, with a mean of 180–220 bp for the three libraries. The libraries contained approximately 40–60% microclones
with unique sequence inserts. More than 30 unique sequence microclones from each library were analyzed by Southern blot hybridization
to demonstrate that they are human specific and were derived from chromosome 18. The human gemomic HindIII fragments hybridized
to each microclone were determined and microclones crosshybridized to rodent species were identified. These region-specific
libraries and the unique sequence microclones from the libraries are useful reagents for (1) isolating hughly polymorphic
microsatellite markers for refined linkage analysis, (2) identifying corresponding YAC, BAC or other clones with large inserts
for contig assembly and high resolution physical mapping, (3) isolating cDNA clones from the dissected region, and (4) convenient
sequencing of the microclones to prepare high density markers and sequence-tagged sites (STSs). Such applications have been
demonstrated in a series of similarly constructed microdissection libraries from other regions of the human genome. 相似文献
119.
Liu J Juo SH Terwilliger JD Grunn A Tong X Brito M Loth JE Kanyas K Lerer B Endicott J Penchaszadeh G Gilliam TC Baron M 《American journal of medical genetics》2001,105(2):189-194
Evidence for linkage between bipolar affective disorder (BP) and 21q22 was first reported by our group in a single large pedigree with a lod score of 3.41 with the PFKL locus. In a subsequent study, with denser marker coverage in 40 multiplex BP pedigrees, we reported supporting evidence with a two-point lod score of 2.76 at the D21S1260 locus, about 6 cM proximal to PFKL. For cost-efficiency, the individuals genotyped in that study comprised a subset of our large pedigree sample. To augment our previous analysis, we now report a follow-up study including a larger sample set with an additional 331 typed individuals from the original 40 families, improved marker coverage, and an additional 16 pedigrees. The analysis of all 56 pedigrees (a total of 862 genotyped individuals vs. the 372 genotyped previously), the largest multigenerational BP pedigree sample reportedly analyzed to date, supports our previous results, with a two-point lod score of 3.56 with D21S1260. The 16 new pedigrees analyzed separately gave a maximum two-point lod score of 1.89 at D21S266, less than 1 cM proximal to D21S1260. Our results are consistent with a putative BP locus on 21q22. 相似文献
120.
慢性癫痫模型动物海马内GABA能神经元的超微结构改变 总被引:10,自引:3,他引:10
用马桑内酯诱发小鼠性性癫痫模型,借助免疫电镜方法对海马的GABA能神经元的超微结构进行观察,可见小鼠海马内GABA免疫反应阳性神经元弥散分布于除锥体细胞层和颗粒细胞层外的各部。慢性癫痫上鼠海马内的GABA免疫反应性神经元胞体和末梢均呈现程度不同的结构损伤,包括线粒体肿胀,细胞器崩解和神经末梢变性,GABA样轴突末梢可与免疫反应阴性的树突构成传出性轴-树突触,也可接受免疫反应阴性轴末梢的传入性轴-轴 相似文献