氟化钠对小鼠全血乙酰明碱酯酶影响的动态观察

氟化钠对小鼠全血乙酰明碱酯酶影响的动态观察吴南屏，赵忠良，康富敏，韩宝红氟中毒患者有明显的乙酰胆碱蓄积症状，表明氟可抑制乙酰胆碱酯酶（ＡＣｈＥ）的活性，但文献报道染氟早期对ＡＣｈＥ活性影响结果不一［１－３］。本实验拟通过动态观察氟对小鼠全血ＡｃｈＥ活...

两种胆碱酯酶试剂的测定评价

本文对国产及进口ＣＨＥ试剂进行评价，两种试剂的组成及不同温度下的稳定性相同，精密度良好（ＣＶ均＜５％），相关系数为０．９８２６～１．０５４．国产试剂吸收峰为４１０ｎｍ，线性１６５００ｕ／Ｌ，回收率９７．５％～１０１．１％，平均９８．６％，正常参考范围６６３４±±２９５６ｕ／Ｌ，进口试剂吸收峰为４０５ｎｍ，线性１４５００ｕ／Ｌ，回收率９９．００％～１０２．３％，平均１００．３％，正常参考范围６６．７９±３０９２ｕ／Ｌ，结果表明，国产ＣＨＥ试剂完全可以替代进口ＣＨＥ试剂。

石杉碱甲治疗增龄相关记忆减退症的双盲对照研究

目的 探讨石杉碱甲胶囊治疗与增龄相关的记忆减退症 (AAMI)的疗效和安全性。方法 采用多中心随机双盲对照研究 ,对照剂为石杉碱甲片剂 (双益平 )。结果 试验组 6 3例服石杉碱甲胶囊 30 μg/d ,对照组 6 1例服石杉碱甲片剂 30 0 μg/d ,疗程 8周。试验过程中进行韦氏记忆量表 (WMS)、简易智力状态检查 (MMSE)、临床疗效总评量表 (CGI)、不良反应症状量表 (TESS)评定及多项实验室检查。两组在治疗后WMS和MMSE评分均显著提高 (P <0 0 5 ) ,两组之间治疗后的WMS和MMSE增分值无显著差异 (P >0 0 5 )。胶囊组总有效率为 6 5 0 8% ,片剂组总有效率为 6 2 30 % ,两组无差异 (P >0 0 5 )。两组的不良反应都很少。结论 石杉碱甲胶囊是一种有效的益智药 ,能改善AAMI,安全性高。

石杉碱甲治疗后阿尔茨海默病患者甲状腺功能的变化

目的:分析阿尔茨海默病与甲状腺功能的关系,观察胆碱酯酶抑制剂石杉碱甲对甲状腺功能的影响并分析可能的机制。方法:阿尔茨海默病组选择2004-08/2006-01解放军总医院第一附属医院的住院及门诊患者27例,健康老龄组为健康志愿者及来本院体检的健康老年人30例。采用放射免疫分析方法,测定阿尔茨海默病患者与健康老龄组血清三碘甲状腺原氨酸(T3)、甲状腺素(T4)、游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺素(TSH)的水平;将阿尔茨海默病患者随机分为石杉碱甲治疗组及吡拉西坦对照组,石杉碱甲治疗组给予石杉碱甲0.1mg,口服,2次/d;吡拉西坦对照组给予吡拉西坦1.2g,口服,3次/d。疗程6个月,在治疗前、治疗后3个月、6个月时分别测定各组甲状腺功能指标水平。结果:阿尔茨海默病组27例和健康老龄组30例均完成各项指标的测定,全部进入结果分析。①阿尔茨海默病组T3、FT3均较健康老龄组显著降低[T3(0.54±0.17),(0.79±0.19)μg/L,t=2.932,P<0.01];[FT3(1.66±0.43),(1.94±0.27)pmol/L,t=5.237,P<0.01]。②重度痴呆患者FT3水平较轻度痴呆患者明显降低[FT3(1.39±0.35),(1.96±0.43)pmol/L,t=2.977,P<0.01];血清FT3水平与痴呆程度呈显著相关(r=0.539,P<0.01)。③石杉碱甲治疗组FT3水平在治疗3月时较治疗前明显增加[(1.82±0.22),(1.67±0.32)pmol/L,t=3.489,P<0.01],治疗6月时较治疗前亦明显增加[(1.79±0.21)pmol/L,t=2.392,P<0.05]。④石杉碱甲组治疗3个月时FT3与吡拉西坦对照组同期相比明显增加[(1.82±0.22),(1.62±0.29)pmol/L,t=2.269,P<0.05],治疗6个月时与对照组同期相比各甲状腺功能指标差异无显著性意义(P>0.05)。结论:甲状腺功能与阿尔茨海默病所致认知功能障碍有关,胆碱酯酶抑制剂石杉碱甲对于认知功能的改善可能归功于通过提高乙酰胆碱的水平,上调甲状腺功能而完成。

IL-1β对大鼠胆碱能神经元的影响

目的 探讨 IL- 1β对大鼠胆碱能神经元的损伤作用及机制。方法 将 IL- 1β缓慢释放颗粒 (IL- 1βpellet)植入大鼠右侧 Meynert核 (nb M,基底核 ) ,复制胆碱能系统受损的动物模型 ;将不同剂量的人重组白细胞介素 - 1β(rh IL- 1β)加入培养大鼠前脑基底神经元中 ,复制 rh IL- 1β诱导神经元损伤模型 ,检测 (乙酰胆碱酯酶 ) Ach E活性和脂质过氧化物 (L PO)含量的变化。结果 IL- 1β损伤侧皮质 Ach E活性较未损伤侧、正常组及假手术组均显著降低 (P <0 .0 1)。 5 0 - 5 0 0 U/ ml rh IL- 1β可引起培养的神经元 Ach E活性明显下降 (P <0 .0 5 ) ;10 0 - 5 0 0 U/ ml rh IL-1β可引起神经元 L PO含量明显升高 (P <0 .0 1)。结论 IL- 1β慢性作用可以造成大鼠中枢胆碱能系统的损害 。

老年痴呆相关蛋白靶标结构与功能关系的分子动力学模拟(英文)

主要运用常规和拉伸分子动力学模拟方法,研究β-淀粉样多肽的构象变化过程、乙酰胆碱酯酶和其抑制剂石杉碱甲的结合与解离过程,以及烟碱乙酰胆碱受体的离子门控构象变化过程,从而为老年痴呆症的病变机理,阐述和设计发现新的抗老年痴呆药物提供线索.

Effect of Transgenic Bt plus CpTI Cotton on Carboxylesterase and Acetylcholinesterase of Cotton Aphid Aphis gossypii

[Objective] The research aimed to assess the effect of transgenic Bt plus CpTI cotton variety SGK321 on carboxylesterase and acetylcholinesterase of cotton aphid Aphis gossypii and provide theoretical basis for studying the biosafety of transgenic cotton.[Method] Cotton aphids were fed with SGK321 and Shiyuan321（normal parental varieties） for over 40 generations.Enzyme activities were compared between cotton aphids feeding on SGK321 for 1,2,3,41,42 and 43 generations with those on Shiyuan321.[Result] The carboxylesterase activity of cotton aphids feeding on SGK321 for 1 generation was significantly higher than those feeding on Shiyuan321.Acetylcholinesterase activity of cotton aphids feeding on SGK321 for 1,2 and 3 generations were significantly higher than those feeding on Shiyuan321 in the same generation.But there was no significant difference of enzyme activity between cotton aphids feeding on SGK321 for a long term and those feeding on parental cotton.[Conclusion] The cotton aphid that feeding on transgenic Bt plus CpTI cotton SGK321 for a long time has adaptivity to SGK321 by regulating the detoxifying enzyme.

Cloning and Sequencing of Two Acetylcholinesterase cDNA Fragments from Cotton Aphid,Aphis gossypii Glover

Two acetylcholinesterase (AChE) genes cDNA fragments,Ag.acel and Ag.ace2,have been cloned from cotton aphid,Aphis gossypii Glover using degenerate primers with RT-PCR technique.Ag.acel gene cDNA fragment is of 282?bp encoding 94 amino acids,and Ag.ace2 gene cDNA fragment is of 264?bp encoding 88 amino acids.Both two putative AChE genes cDNA fragments share numerous similarities with those cloned from other insects.This is the first report of two AChE cDNA fragment sequences in the insect species,which provided the direct evidence of multiple AChE existence in insects.

Morphological changes of cholinergic nerve fibers in the urinary bladder after establishment of artificial somatic-autonomic reflex arc in rats

Objective To establish an artificial somatic-autonomic reflex arc in rats and observe the following distributive changes of neural fibers in the bladder. Methods Adult Sprague-Dawley rats were randomly divided into three groups： control group, spinal cord injury （SCI） group, and reinnervation group. DiI retrograde tracing was used to verify establishment of the model and to investigate the transport function of the regenerated efferent axons in the new reflex arc. Choline acetyltransferase （CHAT） in the DiI-labeled neurons was detected by immunohistochemistry. Distribution of neural fibers in the bladder was observed by acetylcholine esterase staining. Results DR-labeled neurons distributed mainly in the left ventral horn from L3 to L5, and some of them were also CHAT-positive. The neural fibers in the bladder detrusor reduced remarkably in the SCI group compared with the control （P 〈 0.05）. After establishment of the somatic-autonomic reflex arc in the reinnervation group, the number of ipsilateral fibers in the bladder increased markedly compared with the SCI group （P 〈 0.05）, though still much less than that in the control （P 〈 0.05）. Conclusion The efferent branches of the somatic nerves may grow and replace the parasympathetic preganglionic axons through axonal regeneration. Acetylcholine is still the major neurotransmitter of the new reflex arc. The controllability of detrusor may be promoted when it is reinnervated by the pelvic ganglia efferent somatic motor fibers from the postganglionic axons.

Effects of Diazepam on Rat Cholinesterase andMembrane Stability of Red Cell in vitro

The results confirmed that diazepam inhibits the cholinesterase in rat serum,red cell,corpus striatum and diaphragm in vitro,that the higher the diazepam concentration,the stronger the cholinesterase inhibition,and that diazepam is a reversible inhibitor to acetylcholinesterase and diazepam has a stabilizing action on red cell membranes.The role of these effects of diazepam in the treatment of organic phosphate poisoning is discussed

Intestinal pseudo-obstruction:An uncommon condition with heterogeneous etiology and unpredictable outcome

Intestinal pseudo-obstruction (IPO) either acute or chronic is a condition including features of intestinal ileus in absence of mechanical obstruction. Our paper presents such a rare case of idiopathic IPO in a 53-year-old male patient with recurrent episodes of pseudo-obstruction, which were successfully resolved by anticholinesterase agents, motilin agonists or colonic decompression. However, the patient finally underwent total colectomy. Huge colonic dilatation was identified intraoperatorily, while histology showed a neuropathic variant of chronic intestinal pseudo-obstruction. Etiologic mechanisms and current therapeutic methods are reviewed in this paper, which concludes that IPO is a condition in which conservative treatment usually fails. Total colectomy with ileoanal pouch may be the only solution in these situations.

Hirschsprung's disease:Is there a relationship between mast cells and nerve fibers?

AIM:To define the topography of mast cells and their numbers in cases of Hirschsprung's disease(HD)and non-HD,assess neural hypertrophy using imaging software and to study the relationship between mast cells and nerve fibers.METHODS:HE stained sections of 32 cases of chronic constipation in the age group of 0-14 years were reviewed for ganglion cells.AChE staining was performed on frozen sections of colonic and rectal biopsies.Based on their findings cases were divided into HD and non-HD and mast cells stained by toluidine blue were evaluated.Image analysis by computerized software was applied to S-100 stained sections for assessment of neural hypertrophy.RESULTS:Difference between number of mast cells in HD group(mean=36.44)and in non-HD group(mean =14.79)was statistically significant.Image analysis morphometry on S-100 stained sections served as a useful adjunct.The difference between number,size,and perimeter of the nerve fibers between HD and non-HD group was statistically significant.CONCLUSION:Mast cells are significantly increased in HD and their base line values are much higher in Indian children than that reported in Western literature.Their role in HD needs further research.Morphometry of S-100 stained nerve fibers is a useful adjunct to conventional methods for diagnosis of HD.

Development of a Zebrafish Model for Rapid Drug Screening against Alzheimer＇s Disease

Alzheimer＇s disease, the leading cause of dementia in the elderly, is a complex neurodegenerative disorder which leads to a progressive decline in cognitive functions. A rapid screening model is highly demanded for identification and evaluation of novel anti-Alzheimer＇s disease drugs from a large numbers of compounds. Until now, numerous studies utilized zebrafish model for drug discovery. Since aluminum can induce a similar biological activity in zebrafish as in Alzheimer patients, in this study, we developed a novel animal model using 3 to 5 day post-fertilization larval zebrafish by optimizing the doses and duration of aluminum chloride exposure. Six anti-Alzheimer＇s disease drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model. Importantly, Rivastigmine, ThT, Flurbiprofen and AM-117 could increase the value of Dyskinesia Recovery Rate by 53.4-64%, 169.4-200%, 54.5-96% and 70.9-121%, respectively. Rivastigmine, Memantine, ThT, Flurbiprofen, Rosiglitazone and AM-117 improved the value of Response Efficiency by 86.6-175.1%, 28.2-66.6%, 127.2-236.5%, 118.3-323.7%, 26.6-140.8% and 70.2-161.4%, respectively. Our results suggest that the zebrafish model developed in this study could be a useful tool for high throughput screening of potential novel anti-Alzheimer＇s disease leading compounds targeting acetylcholinesterase, N-methyl-D-aspartic acid receptor, γ-secretase, peroxisome proliferator-activated receptor-γand amyloid-β.

Histochemical study of the pre- and postnatal development of acetylcholinesterase in the rat spinal cord

The distribution of acetylcholinesterase(AChE)-positive structures in the developing rat spinal cord was studied with AChE-histochemistry.AChE-positive perikarya were first seen on embryonic day 14(E14) in the ventrolateral portion of the spinal cord.From that time onward.AChE=containing cells appeared gradually in the intermediate gray,dorsal horn and lateral spinal nucleus of the spinal cord in a ventral-to-dorsal,and lateral-to-medial order.No obvious rostral-to-caudal sequence was found.At birth,the distribution pattern of AChE-positive perikarya was basically similar to that in adults.After birth a dramatic increase in the AChE staining intensity extended from postnatal day 5(P5) to postnatal day 21(P21),In addition,two phases of transient AChE staining were observed in the external surface of the dorsal horn from embryonic day 15(E15) to embryonic day 21(E21) and in the marginal layer from embryonic day 21(E21) to postnatal day 14(P14),respectively.

Acute toxicity of organochlorine insecticide endosulfan to the giant freshwater prawn Macrobrochium rosenbergii

Endosulfan, an organochlorine pesticide, is highly toxic and effective at controlling pests in agriculture, horticulture, and public health programs. In this study, static bioassays were used to evaluate the toxicity of endosulfan to freshwater prawns( Macrobrachium rosenbergii) of various lengths(1.5±0.03,4±0.08, and 7±0.06 cm). Additionally, the activities of peroxidase(POD), acid phosphatase(ACP),alkaline phosphatase, acetylcholinesterase(AChE), and Na + /K +-ATPase were analyzed to refl ect the effects of endosulfan exposure. The 96 h LC 50 of endosulfan for prawns 1.5, 4, and 7 cm long were 1.86, 4.53,and 6.09μg/L, respectively, improved tolerance to endosulfan with growth. The POD activities of test organisms exposed to low concentrations of endosulfan were inhibited, indicating the presence of oxygen damaged tissue. Moreover, a notable decrease in AChE activity was observed due to overstimulation of neurotransmission, which might result in abnormal behavior. The effect caused by endosulfan on phosphatase production in the hepatopancreas of prawns 1.5, 4, and 7 cm long was different because the ability of nonspecifi c immune regulation increased with growth. The 96 h LC 50 values obtained in this study could be used in the formulation of water-quality criteria in China. Moreover, the changes in enzymes activities of M. rosenbergii under stress of endosulfan could be applied in the establishment of early warning indicators for bio-safety.

Pros and Cons in Therapeutic Evaluation of Paraoxonase 1 in Nerve Agent Toxicity

PON 1 （Paraoxonase 1） has been proposed as an efficient catalytic bioscavenger to combat against OP （organophosphate） and CWNA （chemical warfare nerve agent） toxicity. Unlike stoichiometric bioscavengers such as butyrylcholinesterase, catalytic bioscavengers are cost effective with the advantage of eliminating all the OPs/CWNAs at low doses. Analysis of catalytic bioscavenger efficacy of PONI showed promising results by various group of researchers. Still, there are large numbers of grey areas which are not addressed so far. One of the major areas of interest is the pharmacokinetic analysis of infused PON 1 in multiple animal models. It is shown that previous studies in mice significantly increased half-life of PONI, while recent studies in guinea pigs from our group showed reduced half-life of PON1. Similar results were reported by other research groups in guinea pigs and non-human primates. The short half-life of exogenously administered PON1 in multiple animal models may be due to poor association of PON1 with its endogenous carrier, high density lipoprotein or lower doses of PON 1 or a reflection of species difference. These observations warrant the significance of thorough pharmacokinetic analysis of infused PON 1 and the development of alternative approaches for successful utility of PON 1 as an efficient medical countermeasure against OP/CWNA toxicity.