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91.
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
92.
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
93.
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
94.
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
95.
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
96.
通过Pro/E和ANSYS建立下颌骨颏部骨折内固定的三维有限元模型,并模拟了各种功能状态下的边界约束。使用计算机图像处理软件对CT扫描图像进行预处理,然后采用自编程序获取下颌骨解剖结构的三维坐标数据,用ANSYS建立下颌骨的三维有限元模型。采用数值化建模软件Pro/E建立小型接骨板-螺钉固定体系统的实体模型。模拟下颌骨颏部正中骨折坚强内固定治疗,建立该内固定治疗的有限元模型。通过改变各组咀嚼肌力的大小,进行了三种咬合状态的边界约束。获得了形态逼真、相似性好的下颌颏部骨折内固定后的三维有限元模型,并模拟了各功能状态下的边界约束,为后期的生物力学分析奠定了基础。 相似文献
97.
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
98.
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
99.
Shuo Dong Jie-Ping Geng Jia-Hua Tong Yu Wu Jin-Ren Cai Guan-Lin Sun Shu-Rong Chen Zhen-Yi Wang Christian-Jacques Larsen Roland Berger Sai-Juan Chen Zhu Chen 《Genes, chromosomes & cancer》1993,6(3):133-139
DNA studies of the translocation t(15;17) in acute promyelocytic leukemia (APL) have shown that the retinoic acid receptor alpha (RARA) gene on chromosome 17 is juxtaposed to the promyelocytic leukemia (PML) gene on chromosome 15. The PML breakpoints have been mapped to 3 clusters: bcr1, bcr2, and bcr3. We have examined the PML breakpoint distribution in a series of 33 Chinese patients with APL Twenty-two patients fell within bcr1, 2 within bcr2, and 9 within bcr3. The primary structure of the reciprocal chromosome translocation joints of one patient and that of their normal counterparts have been determined and compared to those of 2 previously reported cases. These studies revealed possible topoisomerase II cleavage sites close to the breakpoints and suggested implications of DNA attachment sites to nuclear matrix. We propose that these features are relevant to the process of illegitimate recombination generating the translocation. © 1993 Wiley-Liss, Inc. 相似文献
100.
Fertility clinics worldwide routinely produce a large volume of 'waste' follicular aspirate, which is potentially an abundant source of immature ovarian follicles. Current attempts to cultivate these further in vitro to yield viable mature oocytes for fertility treatment have not yet achieved much success. Instead, recent lines of evidence have emerged that are suggestive of a potential stem cell niche within such immature ovarian follicles. The recent discovery of follicular renewal and putative germ-line stem cells within the postnatal mammalian ovary shook the foundations of reproductive biology by challenging the established dogma that mammalian females lose the capacity for germ cell renewal during fetal life, such that a fixed reserve of germ cells (oocytes) enclosed within follicles is endowed at birth. More intriguingly, another recent study in the Drosophila model provided compelling evidence that somatic progenies (nurse cells) of germ-line stem cells had the ability to revert back to the stem-cell-like state. This introduces the exciting possibility that within the mammalian ovarian follicle, similar somatic progenies of germ-line stem cells may also possess a greater intrinsic ability to revert back into functional stem cells. If this is the case, then a favored candidate would be the cumulus/granulosa of immature ovarian follicles, since such cells are true homologues of nurse cells found within the Drosophila ovary. The successful elucidation of a human germ-line stem cell niche within immature ovarian follicles is likely to have huge ramifications in stem cell biology and regenerative medicine. 相似文献