The epididymis is divided into caput, corpus and cauda regions, organized into intraregional segments separated by connective tissue septa (CTS). In the adult rat and mouse these segments are highly differentiated. ...The epididymis is divided into caput, corpus and cauda regions, organized into intraregional segments separated by connective tissue septa (CTS). In the adult rat and mouse these segments are highly differentiated. Regulation of these segments is by endocrine, lumicrine and paracrine factors, the relative importance of which remains under investigation. Here, the ability of the CTS to limit signaling in the interstitial compartment is reviewed as is the effect of 15 days of unilateral efferent duct ligation (EDL) on ipsilateral segmental transcriptional profiles. Inter-segmental microperifusions of epidermal growth factor (EGF), vascular endothelial growth factor (VEGFA) and fibroblast growth factor 2 (FGF2) increased phosphorylation of mitogen activated protein kinase (MAPK) in segments 1 and 2 of the rat epididymis and the effects of all factors were limited by the CTS separating the segments. Microan'ay analysis of segmental gene expression determined the effect of 15 days of unilateral EDL on the transcriptome-wide gene expression of rat segments 1-4. Over 11 000 genes were expressed in each of the four segments and over 2 000 transcripts in segment 1 responded to deprivation of testicular lumicrine factors. Segments 1 and 2 of control tissues were the most transcriptionally different and EDL had its greatest effects there. In the absence of lumicrine factors, all four segments regressed to a transcriptionally undifferentiated state, consistent with the less differentiated histology. Deprivation of lumicrine factors could stimulate an individual gene's expression in some segments yet suppress it in others. Such results reveal a higher complexity of the regulation of rat epididymal segments than that is generally appreciated. (Asian J Androl 2007 July; 9: 565-573)展开更多
目的探讨门静脉高压症术后门静脉系统血栓(portal venous system thrombosis,PVST)形成的原因及其防治措施。方法对132例门静脉高压症患者行脾切除联合贲门周围血管离断术,术中均经胃网膜右静脉置入导管测压并留置导管,其中36例患者(设...目的探讨门静脉高压症术后门静脉系统血栓(portal venous system thrombosis,PVST)形成的原因及其防治措施。方法对132例门静脉高压症患者行脾切除联合贲门周围血管离断术,术中均经胃网膜右静脉置入导管测压并留置导管,其中36例患者(设为试验组,其余设为对照组)术中增加脾静脉近端结扎。术后早期经导管滴注肝素盐水预防血栓,或发生血栓后滴注尿激酶溶栓,出院后口服华法林,使预防和治疗血栓的效果分别达到国际标准化比值(INR)维持于1.5~2.0和2.0~3.0,并直至血小板数量恢复正常。结果术后2周内发生PVST共132例(100%),血栓分布:残余脾静脉血栓132例(100%),门静脉血栓(PVT)39例(29.5%)。在39例PVT中,门静脉主干血栓33例,主干血栓均与残余脾静脉血栓相连,其中15例血栓最大横截面积<50%,14例血栓最大横截面积≥50%,4例为完全性血栓;肠系膜上静脉血栓6例,其中4例合并门静脉主干血栓并与肠系膜上静脉血栓相连,2例存在门静脉分支血栓且肠系膜上静脉血栓与脾静脉血栓相连。39例PVT中,门静脉左支血栓21例,门静脉右支血栓18例;PVT 2处及以上者25例。在试验组36例脾静脉近端结扎的患者中,发生门静脉主干血栓1例(2.8%);在对照组未采用脾静脉近端结扎的96例中发生PVT 38例(39.6%),两者对比差异有统计学意义(P<0.001)。对39例PVT采用抗凝及溶栓治疗,其中33例在术后6个月获得随访并进行CT检查,发现血栓消失、机化再通和海绵样变各23例、7例和3例。结论肝硬化门静脉高压症行脾切除联合贲门周围血管离断术后早期残余脾静脉内易形成血栓,残余脾静脉血栓向门静脉内蔓延是发生术后PVT的主要原因。脾静脉近端结扎的预防效果显著,经胃网膜右静脉留置导管,术后滴注肝素盐水和溶栓剂兼具预防和治疗双重作用,口服华法林效果确切但需检测凝血功能。展开更多
A novel method has been developed to monitor the nucleic acids ligation process. Molecular beacon was employed here to convert the ligation information into fluo-rescence signal quickly and quantitatively. This method...A novel method has been developed to monitor the nucleic acids ligation process. Molecular beacon was employed here to convert the ligation information into fluo-rescence signal quickly and quantitatively. This method pro-vides effective and original approach to researching the dy-namic ligation process and the interactions between nucleic acids and ligase. An analytical method for T4 DNA ligase based on this way has been built up with a linear detection range from 2.3×10-4 U/mL to 0.23 U/mL. It is rapid and sensitive to detect 2.8×10-5 U T4 DNA ligase in 10 min.展开更多
Protein-mediated chromatin interactions can be revealed by coupling proximity-based ligation with chromatin immunoprecipitation.However,these techniques require complex experimental procedures and millions of cells pe...Protein-mediated chromatin interactions can be revealed by coupling proximity-based ligation with chromatin immunoprecipitation.However,these techniques require complex experimental procedures and millions of cells per experiment,which limits their widespread application in life science research.Here,we develop a novel method,Hi-Tag,that identifies high-resolution,long-range chromatin interactions through transposase tagmentation and chromatin proximity ligation(with a phosphorothioate-modified linker).Hi-Tag can be implemented using as few as 100,000 cells,involving simple experimental procedures that can be completed within 1.5 days.Meanwhile,Hi-Tag is capable of using its own data to identify the binding sites of specific proteins,based on which,it can acquire accurate interaction information.Our results suggest that Hi-Tag has great potential for advancing chromatin interaction studies,particularly in the context of limited cell availability.展开更多
文摘The epididymis is divided into caput, corpus and cauda regions, organized into intraregional segments separated by connective tissue septa (CTS). In the adult rat and mouse these segments are highly differentiated. Regulation of these segments is by endocrine, lumicrine and paracrine factors, the relative importance of which remains under investigation. Here, the ability of the CTS to limit signaling in the interstitial compartment is reviewed as is the effect of 15 days of unilateral efferent duct ligation (EDL) on ipsilateral segmental transcriptional profiles. Inter-segmental microperifusions of epidermal growth factor (EGF), vascular endothelial growth factor (VEGFA) and fibroblast growth factor 2 (FGF2) increased phosphorylation of mitogen activated protein kinase (MAPK) in segments 1 and 2 of the rat epididymis and the effects of all factors were limited by the CTS separating the segments. Microan'ay analysis of segmental gene expression determined the effect of 15 days of unilateral EDL on the transcriptome-wide gene expression of rat segments 1-4. Over 11 000 genes were expressed in each of the four segments and over 2 000 transcripts in segment 1 responded to deprivation of testicular lumicrine factors. Segments 1 and 2 of control tissues were the most transcriptionally different and EDL had its greatest effects there. In the absence of lumicrine factors, all four segments regressed to a transcriptionally undifferentiated state, consistent with the less differentiated histology. Deprivation of lumicrine factors could stimulate an individual gene's expression in some segments yet suppress it in others. Such results reveal a higher complexity of the regulation of rat epididymal segments than that is generally appreciated. (Asian J Androl 2007 July; 9: 565-573)
文摘A novel method has been developed to monitor the nucleic acids ligation process. Molecular beacon was employed here to convert the ligation information into fluo-rescence signal quickly and quantitatively. This method pro-vides effective and original approach to researching the dy-namic ligation process and the interactions between nucleic acids and ligase. An analytical method for T4 DNA ligase based on this way has been built up with a linear detection range from 2.3×10-4 U/mL to 0.23 U/mL. It is rapid and sensitive to detect 2.8×10-5 U T4 DNA ligase in 10 min.
基金supported by the National Natural Science Foundation of China(32221005)the Earmarked Fund for CARS(CARS-35)+1 种基金the National Natural Science Foundation of China Outstanding Youth(32125035)Major Project of Hubei Hongshan Laboratory(2021hszd003)。
文摘Protein-mediated chromatin interactions can be revealed by coupling proximity-based ligation with chromatin immunoprecipitation.However,these techniques require complex experimental procedures and millions of cells per experiment,which limits their widespread application in life science research.Here,we develop a novel method,Hi-Tag,that identifies high-resolution,long-range chromatin interactions through transposase tagmentation and chromatin proximity ligation(with a phosphorothioate-modified linker).Hi-Tag can be implemented using as few as 100,000 cells,involving simple experimental procedures that can be completed within 1.5 days.Meanwhile,Hi-Tag is capable of using its own data to identify the binding sites of specific proteins,based on which,it can acquire accurate interaction information.Our results suggest that Hi-Tag has great potential for advancing chromatin interaction studies,particularly in the context of limited cell availability.