非阿片类中药强镇痛剂对大鼠脊髓背角WDR神经元电活动的影响

目的：探讨中药痛必定注射液对正常大鼠及慢性痛模型（CCI）大鼠脊髓背角广动力范围（WDR）神经元晚成分放电的影响，旨在研究中药痛必定注射液的镇痛作用机理。方法：成年SD大鼠20只，随机分成（1）假手术组：大鼠仅暴露坐骨神经未结扎；（2）CCI模型组：实行坐骨神经结扎，制成CCI模型。于术后7-10天，用光热测痛仪测试模型组动物的痛阈，痛阈降低者为模型成功。采用单管玻璃微电极细胞外方法记录正常大鼠及CCI大鼠脊髓背角神经元的诱发放电，根据诱发的背角神经元的反应形式，既有A反应又大鼠有B反应的为WDR神经元，观察并记录给予中药痛必定后2min、4min、8min、15min、30min、60min、120min、180min、240min对正常大鼠及CCI慢性痛大鼠的WDR晚成分放电数、潜伏期及自发放电的影响。     

脊髓背角HCN通道在神经病理性疼痛中的作用

目的:观察脊髓背角超极化激活环核苷酸门控阳离子通道(HCN通道)在坐骨神经缩窄性损伤(CCI)所致的慢性神经病理性疼痛中的作用。方法:8周龄成年雄性SD大鼠48只随机分为6组:(1)sham组(假手术组)、(2)CCI组(鞘内注射生理盐水)、(3)~(6)ZD7288+CCI(鞘内分别注射1,10,30,50μg ZD7288),每组8只。CCI及CCI+ZD7288组大鼠在鞘内置管5 d后行CCI术,术后鞘内给药,每日两次,连续14 d;sham组大鼠不进行鞘内置管,仅游离坐骨神经,不结扎。分别于CCI术前1 d,术后1、3、5、7、10、14 d鞘内给药2 h后测定热缩足潜伏期(TWL);术后第7、14 d处死大鼠,取术侧L4~L6脊髓背角,采用Western Blot技术检测脊髓背角HCN1,3,4及磷酸化蛋白激酶A(P-PKA)表达的变化。结果:大鼠CCI术后即形成稳定的热痛敏,TWL明显缩短;与CCI组相比,鞘内给予HCN通道阻滞剂ZD7288可明显延长CCI大鼠的TWL(P0.05)。Western Blot结果显示,与假手术组相比,CCI组大鼠在术后7、14 d术侧脊髓背角HCN1,3,4及P-PKA表达显著增加(P0.05);鞘内给予ZD7288可显著降低CCI大鼠HCN1,3,4及P-PKA的表达(P0.05)。结论:脊髓背角HCN通道的激活可促进CCI所致的神经病理性痛的发生与维持,HCN通道阻滞剂ZD7288具有良好的镇痛效应,ZD7288的镇痛作用可能与其抑制PKA的活性密切相关。     

坐骨神经损伤大鼠模型鞘内移植神经干细胞后脊髓背角和背根神经节脑源性神经营养因子的表达

背景：坐骨神经损伤模型可测试伤害性的热刺激和机械刺激所引发的痛觉过敏及冷、触觉异常。 目的：观察坐骨神经损伤模型大鼠鞘内移植神经干细胞后脊髓背角和背根神经节脑源性神经营养因子的表达。 方法：72只SD大鼠随机均分为假手术组、对照组和实验组。对照组和实验组制作坐骨神经损伤模型,假手术组仅暴露坐骨神经,不结扎。分别于造模后第3,10天进行鞘内移植,实验组注入30 μL的神经干细胞悬液,空白组和对照组注入30 μL的细胞培养液。 结果与结论：与假手术组相比,对照组和实验组移植后3 d机械痛阈和热痛阈逐渐降低,至移植后7 d降低至最低点(P < 0.01),于移植后21 d恢复至移植前水平;实验组移植后7,14 d机械痛阈和热痛阈较对照组明显上升(P < 0.01)。与对照组相比,假手术组移植后7,14,21 d各组大鼠脑源性神经营养因子的表达呈低水平(P < 0.05);移植后14,21 d,实验组脑源性神经营养因子的表达量高于对照组(P < 0.05)。提示鞘内移植神经干细胞可提高脊髓背角和背根神经节中脑源性神经营养因子的表达。从而抑制了周围神经损伤产生的神经病理性疼痛。 关键词：脑源性神经营养因子;神经干细胞;慢性限制损伤;脊髓背角;背根神经节 doi:10.3969/j.issn.1673-8225.2012.10.031     

鞘内注射MRS2211对慢性坐骨神经结扎大鼠脊髓背角P2X_4受体和P38MAPK表达的影响

目的:观察鞘内注射P2Y13受体阻断剂MRS2211对慢性坐骨神经结扎(CCI)大鼠热痛阈及脊髓背角P2X4受体、P38MAPK表达变化的影响,探讨脊髓P2Y13受体在神经病理性疼痛中的作用。方法:成年SD大鼠随机分为3组(n=8):假手术组、CCI模型组(鞘内注射生理盐水)、MRS2211(100 pmol/L)处理组(CCI+鞘内注射MRS2211)。CCI模型组及MRS2211处理组大鼠均于鞘内置管7 d之后行慢性坐骨神经结扎,连续14 d鞘内注射生理盐水、MRS2211(100 pmol/L),2次/d。分别在术前1 d、术后第1,3,5,7,10,14 d测定给药1h后热缩足潜伏期(TWL);分别在第7,14 d处死大鼠,取脊髓背角做Western Blot检测,观察背角P2X4受体和P38MAPK的表达变化。结果:与假手术组相比,CCI组大鼠第1,3,5,7,10,14 d TWL明显缩短(P0.01);与CCI组相比,MRS2211处理组大鼠术后第1,3,5,7,10,14 d TWL明显延长(P0.05)。Western Blot结果观察到,与假手术组相比,CCI组第7 d P2X4受体表达上调(P0.05),在第14 d P2X4受体表达上调更为明显(P0.05);与假手术组相比,CCI组第7 d P38MAPK表达明显上调(P0.05),在第14 d P38MAPK表达有所下降,但与假手术组相比仍具有统计学意义(P0.05);与CCI组相比,MRS2211处理组大鼠第7 d后背角P38MAPK表达上调明显减弱(P0.05),但在第14 d P38MAPK表达有所上升,但与CCI组相比仍具有统计学意义(P0.05);此外,鞘内注射MRS2211并不影响CCI大鼠背角P2X4受体表达变化。结论:鞘内注射P2Y13受体拮抗剂MRS2211可以明显抑制CCI大鼠热痛敏症状和脊髓背角P38MAPK表达的上调,但并不影响CCI大鼠P2X4受体表达变化。这提示MRS2211可以抑制小胶质细胞P38MAPK活化但不影响小胶质细胞P2X4受体表达,这可能是其在脊髓水平发挥镇痛作用的机制之一。     

鞘内注射p38MAPK抑制剂SB203580对CCI大鼠脊髓背角多种嘌呤受体表达的影响

目的:观察鞘内注射p38MAPK磷酸化抑制剂SB203580对慢性坐骨神经结扎(chronic constriction injury,CCI)大鼠热痛阈及脊髓背角P2X_4、P2X_7、P2Y_(12)、P2Y_(13)受体mRNA表达变化的影响。方法:成年SD大鼠随机分为5组(n=8):假手术组(sham组)、CCI模型组(CCI+鞘内注射0.005%DMSO生理盐水)、CCI+鞘内注射SB203580 1μmol/L组、CCI+鞘内注射SB203580 10μmol/L组、CCI+鞘内注射SB203580 50μmol/L组。CCI模型组及CCI+SB203580处理组大鼠均于鞘内置管7 d后行慢性坐骨神经结扎,连续14 d鞘内注射0.005%DMSO生理盐水或SB203580(1,10,50μmol/L),2次/d,分别在术前1 d、术后第1,3,5,7,10,14 d测定给药1 h后热缩足潜伏期(TWL);并在第3,7,14 d取大鼠脊髓背角进行荧光定量PCR,观察脊髓背角P2X_4、P2X_7、P2Y_(12)、P2Y_(13)受体mRNA的表达变化。结果:CCI术后大鼠即形成稳定的热痛敏,与sham组相比,CCI模型组大鼠术后TWL明显缩短(P0.05);与CCI模型组相比,CCI+SB203580 10,50μmol/L处理组大鼠随药物剂量增加,TWL延长更为明显(P0.05)。荧光定量PCR结果显示:与sham组相比,CCI模型组第3,7,14 d,P2X_4、P2X_7、P2Y_(12)、P2Y_(13)受体mRNA的表达上调(P0.05);给予SB203580 50μmoL/L后,大鼠脊髓背角P2X_7、P2Y_(13)受体mRNA表达有所下调(P0.05),但P2X_4和P2Y_(12)受体mRNA表达没有明显变化。结论:鞘内注射p38MAPK磷酸化抑制剂SB203580可以明显抑制CCI大鼠热痛敏行为学表现,其机制与抑制脊髓背角P2X_7、P2Y_(13)受体mRNA表达有关。这提示脊髓背角小胶质细胞p38MAPK活化后上调P2X_7和P2Y_(13)受体基因转录水平可能是p38MAPK在脊髓水平参与伤害性信息传递的机制之一。     

内质网应激介导的JNK通路在大鼠神经病理性疼痛中的作用

目的:检测内质网应激介导的JNK通路在坐骨神经慢性压迫(CCI)模型大鼠中的作用。方法:SD雄性大鼠随机分为假手术组(sham)和模型组(CCI)。CCI前、后1、3、5、7、14 d测定大鼠的热痛敏和机械痛敏;CCI后14 d,免疫组织化学检测大鼠脊髓背角内葡萄糖调节蛋白78(GRP78)、p-JNK、caspase-3和胶质纤维酸性蛋白(GFAP)的表达;TUNEL法检测脊髓背角内的细胞凋亡。结果:与假手术组相比,模型组的热痛敏和机械痛敏在术后均明显下降,脊髓背角内GRP78、p-JNK、caspase-3和GFAP的表达均增高,凋亡染色阳性细胞数目也较多。结论:内质网应激介导的p-JNK途径参与了神经病理性疼痛模型大鼠脊髓背角内神经元凋亡,可能与星形胶质细胞的活化有关。     

神经干细胞移植治疗坐骨神经半切断神经病理性疼痛的最佳剂量

背景：有研究表明神经干细胞移植治疗神经病理性疼痛有一定效果,但神经干细胞移植数量对神经病理性疼痛的疗效并不确切。 目的：观察不同数量神经干细胞鞘内注射对坐骨神经半切断大鼠神经病理性疼痛及脊髓背角、背根神经节胶质源性神经营养因子表达的影响。 方法：选取1只孕14-16 d SD大鼠制备神经干细胞悬液,分别以细胞数1×103,1×104,1×105,1×106,1×107经坐骨神经半切断伤模型大鼠鞘内注入30 μL,并设置模型组和假手术组作对照。记录术前1 d,术后1,3,7,14,21 d 机械痛及热痛阈;术后7,21 d免疫组织化学与RT-PCR检测同侧脊髓背角及背根神经节胶质源性神经营养因子的表达。 结果与结论：与假手术组比较,其他各组术后1 d 痛阈下降,术后7,14 d最低(P < 0.05);与模型组比较,术后7 d,随着移植神经干细胞的浓度不断增加,神经干细胞1×104,1×105,1×106,1×107组痛阈逐渐升高,脊髓背角及背根神经节胶质源性神经营养因子表达逐渐上调(P < 0.05);术后21 d,神经干细胞1×105,1×106,1×107组痛阈均与术前差异无显著性意义,但其胶质源性神经营养因子表达在神经干细胞1×105组最高(P < 0.05)。说明鞘内注射神经干细胞能缓解大鼠的神经病理性疼痛,注射剂量以1×105神经干细胞效果最佳。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

内质网应激特有的caspase-12在神经病理性疼痛大鼠脊髓背角的表达

目的:检测内质网应激特有caspase-12凋亡途径与神经病理性疼痛模型大鼠脊髓背角神经元凋亡之间的关系。方法:SD雄性大鼠随机分为假手术组(sham)和坐骨神经慢性压迫(CCI)模型组。检测大鼠在手术前及术后21 d的热痛敏和机械痛敏;CCI术后21 d,免疫组织化学检测大鼠脊髓背角内葡萄糖调节蛋白78(GRP78)、caspase-12和caspase-3的表达;TUNEL法检测脊髓背角内细胞的凋亡。结果:与假手术组相比,模型组的热痛敏和机械痛敏在术后均明显下降,脊髓背角内GRP78、caspase-12和caspase-3的表达均增高,凋亡染色阳性细胞数目也较多。结论:坐骨神经慢性压迫模型大鼠脊髓背角内神经元凋亡的途径之一是内质网应激特有caspase-12介导的,可能与神经病理性疼痛的发生、发展有关。     

鞘内注射丹参酮ⅡA对坐骨神经慢性压迫性痛大鼠脊髓背角N-甲基-D-天冬氨酸受体表达的影响

目的:研究丹参酮ⅡA对坐骨神经慢性压迫性痛大鼠脊髓背角内N-甲基-D-天冬氨酸受体2B亚基(NR2B)表达的影响,探讨丹参酮ⅡA的镇痛机制。方法:SD雄性大鼠随机分为假手术组和模型组。模型组又分为生理盐水组和处理组。模型组在手术当日及术后每日在大鼠鞘内注射生理盐水0.1 ml和丹参酮ⅡA 20 mg/kg,连续注射14 d。检测各组大鼠在手术前及术后14 d的机械痛阈和热痛阈;术后第14天,免疫组织化学检测大鼠脊髓背角内NR2B的表达。结果:与假手术组比较,生理盐水组大鼠的机械痛阈和热痛阈明显降低,NR2B的表达增多;与生理盐水组比较,处理组的机械痛阈和热痛阈明显升高,脊髓背角内NR2B的表达下降,差异均有统计学意义。结论:鞘内注射丹参酮ⅡA对坐骨神经慢性压迫性痛模型大鼠的镇痛作用可能与降低脊髓背角内NR2B的表达有关。     

鞘内注射丹参酮ⅡA对坐骨神经慢性压迫性痛大鼠脊髓背角CHOP的影响

目的 :研究丹参酮ⅡA对坐骨神经慢性压迫性痛大鼠脊髓背角CCAAT/增强子结合蛋白同源蛋白(CHOP)和caspase-3表达的影响,探讨丹参酮ⅡA的镇痛机制。方法 :SD雄性大鼠随机分为假手术组和模型组。模型组又分为生理盐水组和处理组,分别在手术当日及术后每日鞘内注射生理盐水0.1 ml和丹参酮ⅡA 20 mg/kg,连续注射14 d。检测各组大鼠在手术前及术后14 d的机械痛阈和热痛阈;术后第14天,免疫荧光组织化学检测大鼠脊髓背角内CHOP和caspase-3的表达。结果 :与假手术组比较,生理盐水组大鼠的机械痛阈和热痛阈降低,CHOP和caspase-3的表达增多;与生理盐水组比较,处理组的机械痛阈和热痛阈升高,脊髓背角内CHOP和caspase-3的表达下降,差异均有统计学意义。结论 :鞘内注射丹参酮ⅡA对坐骨神经慢性压迫性痛模型大鼠的镇痛作用可能与降低脊髓背角内CHOP的表达有关。     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Intestinal helminthiasis in school children in Haiti in 2002

A survey on intestinal helminths in school children was conducted in Haiti in 2002. This first nationwide study involving the entire country was stratified by department according to urban and rural zones using the cluster method. Focusing on elementary school children (n=5792; age range 3 to 20 years), it involved 26 urban and 49 rural schools randomly selected. Stools were preserved in formalin and examined by the Ritchie technique. Thirty-four per cent of stools (1981/5792) tested positive for intestinal helminths with the following parasites identified: Ascaris lumbricoides (27.3%), Trichuris trichiura (7.3%), Necator americanus (3.8%), Hymenolepsis nana (2%), Taenia sp. (0.3%) and Strongyloides stercoralis (0.2%). The helminth prevalence was higher in rural (38.4%) compared to urban areas (30%). There was no significant difference in prevalence by sex and age. The importance of geohelminths changed from one department to another with the highest prevalence found in the Southern department of Grande Anse (73.7%) and the lowest prevalence in the Center department (20.6%). Five out of the country's nine departments had a similar prevalence varying from 25.5% to 28.2%. Intestinal helminthic polyparasitism was observed in a percentage of infested school children comprise between 3.4% and 28.6% according in relation to the geographical area. A program to fight against geohelminths in school children should be initiated as a public health priority. Albendazole is the drug of choice. Frequency of drug distribution should be based on the prevalence of geohelminths in each department.     

Experience in adult population in dengue outbreak in Delhi

A dengue outbreak has recently hit the Indian capital. We studied the clinical profile of adult patients. Five hundred and sixty patients of dengue infection were admitted in a specially created ward according to the criteria laid down by WHO. Haematemesis (28.28%), epistaxis (26.78%) and malena (14.28%) were some of the common presentations. Similarly lymphadenopathy, especially cervical (30.89%), palatal rashes (26.96%) and hepatomegaly (23.75%) were the most commonly encountered findings on physical examination. Most of the cases were of dengue fever with haemorrhage and only 2.5% cases were classified under dengue haemorrhagic fever or dengue shock syndrome. The average hospital stay was 3.4 days but only 9.8 hours in the eleven patients who died, suggesting their late arrival in preterminal situation giving little time for resuscitation. Thrombocytopenia was not a feature and only 12.85% patients had platelet count less than 70,000/cmm. Most of the patients who were admitted with thrombocytopenia, showed normalization in their platelet counts in next few days. Serological examination demonstrated evidence of recent dengue infection in 41.17% patients. Few patients required blood or platelet concentrate transfusion. Eleven patients died, three due to DIC, one of intracranial haemorrhage and seven due to massive gastric haemorrhage. Rest of the patients recovered completely. Thus we can conclude that recent outbreak in Delhi was of dengue fever with haemorrhage and mortality was very low in patients who came early to the hospital.     

Changes in the hypothalamic nuclei in in experimental allergy

Summary In rabbits subjected to prolonged sensitization and in which the Arthus phenomenon was induced there was a marked reaction of the hypothalamic nuclei. Staining by Gomori's method indicated a cellular swelling, loss of granules, and protoplasmic vacuolization in the supraoptic nucleus. There was a considerable increase in the size of the cross-sectional area of the cells. The same effects were much less well shown in the paraventricular nucleus. These results show that marked signs of increased neurosecretion developed in the animals at the height of the Arthus phenomenon.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten Éksperimental'noi biologii i Meditsiny, Vol. 55, No. 4, pp. 110–113, April, 1963.     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.Originally presented at ECCP 93.