Histochemical studies on the spinal cord of squirrel monkey (Saimiri sciureus)

Summary Detailed histochemical observations have been made on the various components of the spinal cord. The gray matter showed much greater enzymatic activity than the white matter. The neuropil and substantia gelatinosa showed mild G6PD, AChE and BChE activity; moderate SDH, LDH, SE and AC activity; and strong AP, ATPase, and AMPase activity. The pia-arachnoid showed moderate to strong activity of dehydrogenases and phosphatases. The cytoplasm of the neurons and neuronal processes gave moderate to very strong reaction for most of the enzymes except BChE. The areas of synaptic endings show the presence of oxidative enzymes, phosphorylases, ATPase and AMPase. It has been interesting to note that phosphorylases, which have been mostly observed in the cytoplasm, nucleoli and synaptic areas of various neurons and in the neuropil, also show intensely strong reactions in the nuclei of ependymal cells lining the central canal as well as in nuclei of some glial cells.List of Abbreviations used AC Acid phosphatase - AChE Acetyl cholinesterase - AK Alkaline phosphatase - AMPase Adenosine monophosphatase (5 nucleotidase) - AP Amylophosphorylase - ATPase Adenosine triphosphatase - BChE Butyrylcholinesterase - G6PD Glucose-6-phosphate dehydrogenase - LDH Lactic dehydrogenase - MAO Monoamine oxidase - PAS Periodic acid Schiff - SDH Succinic dehydrogenase - SE Simple esterases - UDPG Uridinedephosphoglucose glycogen transferase T.R. Shanthaveerappa in previous publications     

PTPN22在溃疡性结肠炎中的表达及其临床意义

背景:溃疡性结肠炎(UC)的发生、发展与T细胞过度活化有关,蛋白酪氨酸磷酸酶非受体型22(PTPN22)参与T细胞活化的负性调节。目的:探讨PTPN22在UC中的表达及其临床意义。方法:纳入2010年7月～2012年7月武汉市中心医院、武汉大学中南医院就诊的UC患者60例,同时纳入35例IBS患者作为对照组。采用实时定量PCR法检测患者肠黏膜PTPN22 mRNA表达水平。采用全自动红细胞沉降系统分析仪检测血清ESR水平。采用速率散射比浊法检测血清CRP水平。结果:活动期UC患者肠黏膜PTPN22 mRNA表达水平与缓解期UC患者和对照组相比显著升高(P=0.007;P=0.021)。UC患者肠黏膜PTPN22 mRNA表达水平与血清ESR和CRP水平呈正相关(r=0.63,P=0.005;r=0.58,P<0.01)。UC患者疾病严重程度和病变范围与肠黏膜PTPN22 mRNA表达水平呈正相关(r=0.51,P<0.01;r=0.44,P<0.01)。中重度UC患者肠黏膜PTPN22 mRNA表达水平与轻度UC患者相比显著升高(P=0.005),病变位于广泛结肠者肠黏膜PTPN22 mRNA表达水平与病变位于左半结肠和直肠者相比显著升高(P=0.029;P=0.008)。结论:PTPN22 mRNA表达水平与UC疾病活动性、严重程度和病变范围相关。PTPN22可能在UC发病机制中发挥重要作用。     

持久饥饿对涡虫转氨酶、磷酸酶、蛋白水解酶和增殖细胞核抗原的影响

目的探讨持久饥饿对涡虫转氨酶、磷酸酶、蛋白水解酶和增殖细胞核抗原(PCNA)的影响。方法采用全自动生化分析仪测定转氨酶的活性变化,用复性电泳技术分析磷酸酶和蛋白水解酶的变化,用半定量RT-PCR技术分析PCNA mRNA的表达变化。结果饥饿过程中谷丙转氨酶和谷草转氨酶的活性显著升高,是对照水平的10倍以上,恢复喂食后逐渐降低到正常水平;碱性磷酸酶(ALP)的活性在饥饿过程中明显减弱,而酸性磷酸酶(ACP)活性却显著增强;受饥饿影响,140kD和40kD蛋白水解酶的活性均明显增加;然而饥饿过程中PCNA的表达没有明显变化,说明饥饿对该基因的影响很小。结论持久饥饿对涡虫的生理和生化代谢有重要影响,酶活性的变化反应了涡虫适应持久饥饿的能量代谢变化。     

酪氨酸磷酸酶PRL-3单抗的制备及特性鉴定

 目的 制备并鉴定抗肝再生磷酸酶-3（Phosphatase of Regenerating Liver-3,PRL-3）单克隆抗体,为临床检测和以 PRL-3为靶点的肿瘤治疗提供可能。方法 运用杂交瘤融合技术制 备PRL-3单克隆抗体,通过Western blot检测和免疫沉淀鉴定其与 原核及真核PRL-3蛋白的反应性; 重组并诱导表达PRL-3的6个截短 体,Western blot分析单抗结合的大致抗原表位。结果 共得到9 株单抗,3株与PRL-1、PRL-2和PRL-3均反应,6株(9D8、9E2、9F4 、11B2、4D3和4D10)只与PRL-3反应,其中4株特异性单抗可以与哺 乳动物细胞表达的PRL-3反应;单抗9D8、9E2、9F4和11B2 可以和 PRL-3羧基末端（162-173位氨基酸）的短肽结合;4D3和4D10与中 间部（69~95位氨基酸）的短肽结合。结论 抗体9D8、9E2、9F4、 11B2、4D3和4D10特异性强、亲和力高,为临床检测提供了可靠工 具,并为进一步的功能研究奠定了基础。     

PTEN与信号转导及肿瘤

ＴＥＮ[1] (ｐｈｏｓｐｈａｔａｓｅａｎｄｔｅｎｓｉｎｈｏｍｏｌｏｇｙｄｅｌｅｔｅｄｏｎｃｈｒｏｍｏｓｏｍｅｔｅｎ)又名ＭＭＡＣ1 [2 ] (ｍｕｔａｔｅｄｉｎｍｕｔｉｐｌｙａｄａｎｃｅｄｃａｎｃｅｒ 1 )和ＴＥＰ1 [3 ] (ＴＧＦ -βｒｅｇｕｌａｔｅｄａｎｄｅｐｉｔｈｅｌｉａｌｃｅｌｌ -ｒｉｃｈｅｄｐｈｏｓｐｈａｔａｓｅ 1 ) (以下均称为ＰＴＥＮ) ,是 1 997年由 3个研究小组先后发现的一个具有双特异磷酸酶活性的抑癌基因。ＰＴＥＮ基因异常广泛存在于人类多种恶性肿瘤 ,如恶性神经胶质瘤、前列腺癌、子宫内膜癌、黑色素瘤等…     

人钙调磷酸酶-B真核表达质粒的构建

目的 为研究钙振荡现象,构建人钙调磷酸酶-B(CNB)基因的真核表达质粒.方法 采用RT-PCR技术,从MCF-7细胞中扩增出人CNB基因序列;利用分子克隆技术将其插入到真核表达载体pcDNA3.0中,并进行酶切鉴定和DNA测序.结果 琼脂糖凝胶电泳分析显示RT-PCR产物为预期分子量大小,酶切鉴定图谱正确,DNA测序结果经核实完全符合.结论 成功构建人CNB基因的真核表达质粒,为进一步研究钙振荡现象打下了基础.     

miR-96的表达对肝细胞癌细胞迁移和侵袭的影响

目的:探讨miR-96在肝细胞癌(HCC)细胞中的表达及作用。方法:用qRT-PCR测定miR-96在不同HCC细胞系(HepG2、7721、huh7)及正常肝细胞系L02中的表达;将HepG2细胞分别转染miRNA随机序列(阴性对照组)、miR-96模拟物(miR-96模拟物组)和miR-96抑制物(miR-96抑制物组)后,用细胞划痕实验及Transwell细胞侵袭实验分别检测细胞迁移及侵袭能力,qRT-PCR及Western blot分别测定PTPN9 mRNA及蛋白的表达。结果:miR-96在各肝癌细胞系中的相对表达量均明显高于其在正常肝细胞系L02的相对表达量(均P0.01)。与阴性对照组比较,细胞划痕愈合率在miR-96模拟物组明显升高,而在miR-96抑制物组明显降低(均P0.05);侵袭细胞数在miR-96模拟物组明显增多,而在miR-96抑制物组明显减少(均P0.05);PTPN9 mRNA与蛋白相对表达量在miR-96模拟物组均明显下调,而在miR-96抑制物组均明显上调(均P0.05)。结论:miR-96在HCC细胞中表达升高,并可能通过下调PTPN9表达促进HCC细胞的迁移与侵袭。     

实验性兔脑水肿脑组织的组织化学研究——Ⅱ.大脑磷酸酶的动态变化

观察伤寒内毒素引起实验兔脑水肿时脑内磷酸酶的动态变化,用自动图象分析仪作定量测定,结果脑毛细血管壁和脉络丛的三磷酸腺苷酶活性减低;大脑皮质神经细胞内酸性磷酸酶活性明显减弱;注伤寒内毒素后1小时脉络丛的碱性磷酸酶活性升高,提示伤寒内毒素引起的脑水肿为混合型(血管源性和细胞毒性脑水肿)。     

Tooth development in Ambystoma mexicanum: phosphatase activities, calcium accumulation and cell proliferation in the tooth-forming tissues

Prerequisites of tooth formation, cell proliferation in the tooth-forming tissues, calcium accumulation and the enzymatic activities of alkaline (ALP) and acid phosphatases (ACP) were investigated by immunohistochemical and histochemical methods in various developmental stages of the Mexican Axolotl, Ambystoma mexicanum. During the growth of replacement teeth, the tooth-forming tissues continually recruit cells from the surrounding regions. The basal layer of the oral epithelium, the dental lamina and sometimes even the outer enamel epithelium provide cells for the differentiated inner enamel epithelium, in which the active ameloblasts are localized. The differentiating odontoblasts are derived from proliferating cells situated basally to the replacement teeth in the mesenchymal tissue. When differentiation has started and the cells have become functional, proliferative activity can no longer be observed. Calcium is accumulated close to the site of mineralization in the inner enamel epithelium and in the odontoblasts as it is in mammals, elasmobranchii and teleostei. The activities of ACP and ALP related to the mineralization of the replacement teeth are separated spatially and not sequentially as they are in mammals. However, the results indicate a similar function of these enzymatic components in relation to tooth formation and maturation of mineral deposition. Most of the substantial processes related to tooth formation reported from other vertebrates occur in a manner similar to that in Ambystoma mexicanum, but there also seem to be basic mechanisms present that are realised in a unique way in this urodele.     

Assessment of biochemical components and enzyme activities in the estuarine crab Scylla tranquebarica from naphthalene contaminated habitants

To document the impact of naphthalene, a comparative toxicological research was performed in the estuarine crab Scylla tranquebarica habitant of Ennore creek (polluted site) and Kovalam creek (pristine site) for a period of 6 months. The biochemical constituents, such as protein, carbohydrate, lipid and enzyme activities like acid and alkaline phosphatase, aspartate and alanine transaminase were analysed in hepatopancreas, heamolymph and ovary of the female crabs collected from the two creeks. The results revealed that there was a significant measurable difference in these parameters in the tissues of crabs sampled from the polyaromatic hydrocarbon (PAH) polluted site (Ennore creek) when compared with the reference site (Kovalam creek). In addition naphthalene was detected in gill, hepatopancrease, heamolymph and ovary of the crabs sampled from the polluted creek. The results indicate that the indicators selected for toxic impact in S. tranquebarica may be related to the uptake of naphthalene in the tissues examined and support the feasibility of employing these types of analyses as biochemical biomarkers for PAH contaminant-exposed organisms.