8Hz次声作用后大鼠脑热休克蛋白70的表达

目的探讨次声作用后大鼠脑的热休克蛋白７０（ｈｅａｔｓｈｏｃｋｐｒｏｔｅｉｎ７０,ＨＳＰ７０）的表达。方法大鼠暴露于８Ｈｚ的次声,２ｈ／ｄ,点杂交检测次声作用１次后多器官ＨＳＰ７０ＭＲＮＡ平;免疫组织化学方法检测次声作用１、３、７、１４、２１ｄ后脑ＨＳＰ７０的表达和分布。结果次声作用后脑、心、肝、肺等器官ＨＳＰ７０ｍＲＮＡ明显增加,脑中有２４个部位出现ＨＳＰ７０阳性神经元,且随声压增强、作用时间延长,阳性表达增强。结论这些部位对次声敏感,次声的作用效应与声压、作用时间有关。     

次声对脑组织血栓素A2,前列环素代谢的影响

目的 探讨次声作用后脑皮层组织血栓素Ａ２（ＴＸＡ２）、前列环素（ＰＬＩ２）代谢改变及及代谢性谷氨酸受体拮抗剂ＭＣＰＧ的作用。方法 ４０只ＳＤ大鼠随机分为正常对照,次声作用１次、７次、１４次及代谢性谷氨酸受体拮抗剂ＭＣＰＧ治疗５组,采用第四军医大学研制的次声压力仓,用８Ｈｚ、１２０ｄＢ的次声按规定次数。每次作用２ｈ。采用蛋白定量和放免法行脑ＴＸＡ２、ＰＬＩ２稳定代谢产物血栓素Ｂ２（ＴＸＢ２）及６－酮     

人FⅧ基因高效表达质粒的构建及其在Cos-7细胞中的表达

目的：构建人ＦⅧ真核表达质粒——ｐＡｄＣＭＶＬｉｎｋＦⅧＤＢ并观察其在Ｃｏｓ－７细胞中的表达活性。方法：采用缺失大部分Ｂ结构域的人凝血因子ⅧｃＤＮＡ（ＦⅧＤＢ），长度为４．６ｋｂ，将其插入含腺病毒序列表达质粒——ｐＡｄＣＭＶＬｉｎｋ１，构建了ＦⅧ真核表达质粒——ｐＡｄＣＭＶＬｉｎｋＦⅧＤＢ，用脂质体介导法将其转染Ｃｏｓ－７细胞，转染后２４，４８，７２小时分别用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）、ＥＬＩＳＡ法及一期法测定培养细胞中ＦⅧＤＢｍＲＮＡ及ＦⅧ的含量与活性。结果：ＲＴ－ＰＣＲ法可检测到ＦⅧＤＢｃＤＮＡ转录的ｍＲＮＡ，ＥＬＩＳＡ法测得７２小时后ＦⅧ的含量为每２４小时１８ｎｇ／１０６细胞，一期法测得活性为每２４小时０．６Ｕ／１０６细胞，相当于正常人血浆中１００μｇ／ＬＦⅧ所产生的凝血活性的６０％。结论：所构建的ＦⅧ真核表达质粒在Ｃｏｓ－７细胞中具有一定的表达活性。     

Fos蛋白和Jun蛋白在侧位液压冲击脑损伤大鼠大脑皮质的表达

目的：研究大鼠中度（０．２ＭＰａ）侧位液压冲击脑损伤时大脑皮质立即早期基因ｃ－ｆｏｓ和ｃ－ｊｕｎ表达产物Ｆｏｓ蛋白和Ｊｕｎ蛋白的变化规律。方法：雄性ＳＤ大鼠,随机分为正常对照物、手术对照组和损伤组。损伤组动物均给以０．２ＭＰａ液压冲击脑损伤,按冲击后处死时间不同再分为５、１５、３０、６０、１２０、２４０、４８０和７２０分钟组。应用免疫组织化学方法观察Ｆｏｓ和Ｊｕｎ蛋白在大脑皮质的表达特点。结果：冲击后３０分钟,双侧大脑皮质Ｆｏｓ阳性细胞数逐渐增多,冲击后７２０分钟达高峰。Ｆｏｓ阳性细胞面积在冲击后１２０分钟逐渐增大,７２０分钟达高峰;冲击后６０分钟双侧大脑皮质Ｊｕｎ阳性细胞数逐渐增多。冲击后１２０分钟阳性细胞面积逐渐增大,冲击后７２０分钟阳性细胞数和面积均达高峰。结论：中度侧位液压冲击脑损伤后Ｆｏｓ蛋白和Ｊｕｎ     

酸枣仁汤对ＲＥＭ 睡眠剥夺老年大鼠脑神经 细胞凋亡及ｃ ｆｏｓ基因表达水平的影响

目的　观察酸枣仁汤对快动眼（ＲＥＭ）睡眠剥夺导致的老年大鼠脑皮层和海马部位神经细胞凋亡及ｃｆｏｓ 基因表达Ｆｏｓ蛋白水平变化的影响。方法　将自然衰老２４月龄Ｗｉｓｔａｒ大鼠随机分成空白对照组（等容生理盐水）， 老年ＲＥＭ睡眠剥夺组（等容生理盐水），阳性对照组（舒乐安定０１８ｍｇ／（ｋｇ·ｄ）），酸枣仁汤低、高剂量组（１２９６， ２５９２ｇ／（ｋｇ·ｄ））。各组灌胃给药２周后，除空白对照组外其余各组用自制改良多平台法剥夺大鼠睡眠４８ｈ制作老年大 鼠ＲＥＭ睡眠剥夺模型。通过原位末端标记法（ＴＵＮＥＬ）测定各组大鼠大脑皮层和海马部位神经细胞凋亡水平，用免疫 组化法检测ｃ ｆｏｓ基因表达Ｆｏｓ蛋白的水平变化，探讨酸枣仁汤对老年失眠引起的脑神经损害及ｃ ｆｏｓ基因表达变化的影 响。结果　与空白对照组比较，老年ＲＥＭ睡眠剥夺组大鼠脑皮层和海马部位神经凋亡细胞增多，Ｆｏｓ蛋白表达明显增 高。与老年ＲＥＭ睡眠剥夺组比较，酸枣仁汤组脑神经凋亡细胞及Ｆｏｓ蛋白表达明显下降，但仍高于正常对照组。结论 　ｃ ｆｏｓ基因很可能参与了老年大鼠ＲＥＭ睡眠剥夺后的细胞凋亡过程，酸枣仁汤可能是通过抑制脑组织ｃ ｆｏｓ基因表达 上调，抑制神经细胞凋亡而发挥脑保护作用。     

外周炎性刺激条件下大鼠脊髓背角中PPTAmRNA和Fos阳性神经 …

以鹿角莱胶（ＣＡＲ）注射到大鼠后爪皮下作为外周炎性刺激模型，用原位杂交结合免疫组化染色观察了ＢＰＴＡｍＲＮＡ在脊髓背角神经元中的表达变化及其与Ｆｏｓ蛋白阳性神经元的关系。结果显示：ＰＰＴＡｍＲＮＡ阳性神经元主要位于脊髓背角柜Ⅰ、Ⅱ层和Ⅴ、Ⅵ层。ＣＡＲ注射后ＰＰＴＡｍＲＮＡ阳性神经元的娄得与对照侧相比明显增多，ＣＡＲ刺激也诱导了Ｆｏｓ蛋白在脊髓背角神经元中的表达。Ｆｏｓ阳性神经元主要位于刺激侧脊髓神     

双倍剂量吸入激素及单剂量吸入激素加茶碱治疗哮喘疗效比较

目的：比较吸入双倍剂量二丙酸倍氯米松（ＢＤＰ）和吸入单剂量ＢＤＰ加小剂量茶碱对哮喘的作用。方法：吸入单剂量ＢＤＰ４００μｇ／ｄ,连续２８ｄ后症状不能控制者,测第１秒用力呼气量（ＦＥＶ１）。增加ＢＤＰ到８００μｇ／ｄ或增加用氨茶碱６００ｍｇ／ｄ,第７天、２８天分别测峰值呼气流速（ＰＥＦ）、第１ｓ用力呼气容积。结果：两组ＰＥＦ７ｄ后均有改善,统计学无显著性差异。ＦＥＶ１２８天后均有明显增加,有显著性差     

次声对大鼠视网膜Occludin蛋白表达改变的影响

目的：探讨次声对大鼠视网膜Occludin蛋白表达变化的影响，旨在从分子水平上探讨次声作用致大鼠血-视网膜屏通透性改变的机制。方法：每组6只大鼠，8Hz。130dB次声暴露1，7，14，21d，对照组亦每日置于次声舱中2h，但不接受次声暴露。于各时间点次声暴露后2h内取视网膜组织标本，用于Western-blot分析，并对蛋白表达变化行统计学分析。结果：次声暴露后导致大鼠视网膜组织Occludin蛋白表达不同程度的下降，但与暴露时间无线形关联。结论：次声导致大鼠血-视网膜屏障通透性的改变，至少部分是由于使视网膜Occludin蛋白表达下降。     

一种新的检测抗双链DNA抗体的斑点免疫结合试验

以纯化的不含单链ＤＮＡ的大肠杆菌质粒双链ＤＮＡ（ｄｓ一ＤＮＡ）作为抗原，经生物素化后，结合于包被了亲和素的硝酸纤维素膜上，建立了一种新的检测抗ｄｓ一ＤＮＡ抗体的斑点免疫结合试验（ＤＩＢＡ）。对１２０份临床血清标本的检测结果显示：４１例未经选择的系统性红斑狼疮（ＳＬＥ）患者有２２例阳性，占５４％，其中活动性ＳＬＥ１９例，阳性１４例，占７４％；２９例其他自身免疫病患者仅１例阳性；５０例正常人全部阴性。与短膜虫免疫荧光法（ＣＬ，ＩＦＡ）和酶联免疫吸附试验（ＥＬＩＳＡ）试剂盒比较的结果为：ＤＩＢＡ的敏感性优于ＣＬ，ＩＦＡ，特异性优于ＥＬＩＳＡ试剂盒。ＤＩＢＡ快速、简便、可靠，可作为检测抗ｄｓ一ＤＮＡ抗体的常规方法。     

胆固醇调节元件结合蛋白

胆固醇调节元件结合蛋白（ＳＲＥＢＰｓ），是一类能与胆固醇调节元件１（ＳＲＥ－１）结合的“碱性螺肇－环－螺旋－亮氨酸拉链”蛋白，在细胞内缺乏胆固醇的情况下，ＳＲＥＢＰｓ通过和ＳＲＥ－１的结合，激活具有ＳＲＥ－１的基因，如低密度脂蛋白受体（ＬＤＬＲ）基因、羟甲基戊二酸单酰ＣｏＡ合酶（ＨＭＧＣｏＡｓｙｎｔｈａｓｅ）基因，发挥调节基因翻译的作用，从而维护细胞内胆固醇含量的平衡。对ＳＲＥＢＰｓ的研究，可能会     

次声作用对大鼠记忆功能及隔内侧核和斜角带核胆碱能神经元表达的影响

目的　探讨次声作用对大鼠记忆功能及斜角带核和隔内侧核胆碱能神经元表达的影响。方法　记忆保持SD大鼠接受16Hz,90dB或130dB次声照射,2h/次     

8Hz次声对大鼠体重及胃十二指肠5-HT表达的影响

目的探讨 8Hz ,90dB、13 0dB次声对SD大鼠体重的影响及其可能机制。方法 3 0只雄性SD大鼠按体重随机分为对照组 ,8Hz、90dB及 8Hz、13 0dB组 3组。实验组分别暴露于 8Hz、90dB或 8Hz、13 0dB次声仓中 ,每日作用时间 2小时 ,共 42天。对照组每日置次声仓中 ,但不予次声作用 ,所有动物每 3天称体重 1次。另 75只随机分为对照组和 8Hz ,90dB、13 0dB的 7、14、2 1、2 8、3 5天组共 15组 ,每组 5只 ,按组别予以不同时间及强度的次声作用 ,对照组每日置次声仓中 ,但不予次声作用。最后一次从次声仓取出后立即取胃及十二指肠 (包括体重实验组各 5只 ) ,免疫组织化学染色显示其 5 羟色胺 (5 HT)含量。光学显微镜下计数胃窦及十二指肠 5 HT阳性细胞数。结果实验组大鼠体重增长均较对照组缓慢 (P =0 0 0 0 ) ,其中 13 0dB组对大鼠体重增长较 90dB组增长缓慢 (P =0 0 0 0 ) ;实验组动物胃窦及十二指肠 5 HT含量较对照组增多 ,以 90dB的 3 5天和 13 0dB的 2 8天明显 (P <0 0 1)。结论 8Hz、90dB及 8Hz、13 0dB次声对雄性SD大鼠体重的增长有抑制作用 ,其机制可能与胃及十二指肠 5 HT增多有关。     

乙状结肠痛对大鼠中缝背核Fos-LI和NADPH-d阳性神经元表达影响的实验研究

目的：探讨中缝背核还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)阳性神经元是否参与调控大鼠乙状结肠痛。方法：采用Fos免疫组织化学、NADPH-d组织化学及Fos／NADPH-α双标技术，观察乙状结肠痛大鼠中缝背核NADPH—d阳性神经元和Fos样免疫反应蛋白(Fos-LI)的变化。结果：乙状结肠痛刺激后，中缝背核内NADPH-d阳性神经元个数和染色深度增加；中缝背核Fos—LI明显增加；中缝背核内有Fos-LI／NADPH-d双标神经元的表达，约占NADPH-d神经元的11％，与生理盐水对照组相比有显著性差异。结论：中缝背核NADPH-d阳性神经元可能参与对乙状结肠痛刺激的信息传递。     

次声不同时间作用后对大鼠大脑皮质超微结构的影响

目的进一步探讨次声对夫鼠大脑作用阈值及量效关系，观察次声不同时间作用后对大鼠脑皮质超微结构的影响。方法大鼠暴露于16Hz、90dB的次声，2h／d，分组作用7，14，21，28，35d，透射电镜下观察各组动物于次声暴露结束后2h、3d、7d等不同时间点脑顶叶皮质超微结构的变化。结果16Hz、90dB次声，2h／d，作用7，14d后脑皮质超微结构无明显变化；作朋21，28，35d后脑皮质超微结构出现变性改变，从21d开始随作用时间延长变性损伤加重；各组变性损伤随作用后时间延长可恢复正常。结论16Hz、90dB次声作用一定次数后大鼠脑皮质超微结构可见变性改变，随作用后时间延长可逐渐恢复正常。     

8 Hz次声对大鼠海马和颞叶皮层5-HT表达的影响

目的研究 8Hz ,90dB、10 0dB、13 0dB次声对SD大鼠海马及颞叶皮层 5 HT表达的影响。方法 14 0只雄性SD大鼠随机分为正常对照组及 8Hz ,90dB、10 0dB、13 0dB次声作用 1、7、14、2 1、2 8、3 5、42d组共 2 8组 ,每组 5只。试验组按组别分别暴露于次声仓中 ,每日 2h ;对照组亦暴露于次声仓 ,每天 2h ,但不予次声作用。最后一次次声作用结束后立即取脑组织并进行 5 HT免疫组织化学染色 ,光学显微镜下观察海马及颞叶皮层 5 HT表达的变化。结果次声作用组大鼠脑组织海马及颞叶皮层 5 HT阳性纤维均较对照组明显减少 (均为P <0 .0 1) ,90dB组、10 0dB组以 2 8d时减少最为明显 ,且 10 0dB组的阳性纤维数量较 90dB组少 ;13 0dB组以 2 1d时减少最为明显。各实验组的阳性纤维数量在此后均有所增加。结论 8Hz ,90dB、10 0dB和 13 0dB次声可引起大鼠海马及颞叶皮层 5 HT表达减少 ,其变化规律与次声作用参数有关 ,相同作用时间下 ,13 0dB组的变化较 10 0dB及 90dB组明显。     

Perinatal asphyxia exerts lifelong effects on neuronal responsiveness to stress in specific brain regions in the rat.

BACKGROUND: Perinatal asphyxia (PA) causes irreversible damage to the brain of newborns and can produce neurologic and behavioral changes later in life. To identify neuronal substrates underlying the effects of PA, we investigated whether and how neuronal responsiveness to an established stress challenge is affected. METHODS: We used Fos expression as a marker of neuronal activation and examined the pattern of Fos expression in response to acute swim stress in 24-month-old rats exposed to a 20-minute PA insult. RESULTS: Swim stress produced a similar pattern of Fos expression in control and asphyxiated rats in 34 brain areas. Asphyxiated rats displayed a higher number of stress-induced Fos-positive cells in the nucleus of the solitary tract, parabrachial nucleus, periaqueductal gray, paraventricular hypothalamic nucleus, nucleus accumbens, caudate-putamen, and prelimbic cortex. No differences in the Fos response to stress were observed in other regions, including the locus ceruleus, amygdala, hippocampus, or septum. CONCLUSION: These data provide functional anatomic evidence that PA has lifelong effects on neuronal communication and leads to an abnormal, augmented neuronal responsiveness to stress in specific brain areas, particularly in the main telencephalic target regions of the mesencephalic dopamine projections, as well as in a functionally related set of brain regions associated with autonomic and neuroendocrine regulation.     

Induction of Fos protein-like immunoreactivity in the trigeminal spinal nucleus caudalis and upper cervical cord following noxious and non-noxious mechanical stimulation of the whisker pad of the rat with an inferior alveolar nerve transection

After transection of the inferior alveolar nerve (IAN: the third branch of the trigeminal nerve), the whisker pad area, which is innervated by the second branch of the trigeminal nerve, showed hypersensitivity to mechanical stimulation. Two days after IAN transection, the threshold intensity for escape behavior to mechanical stimulation of the ipsilateral whisker pad area was less than 1.0 g, a sign of allodynia, and returned to the preoperative level (preoperative threshold: 52.0 g) at 32 days after surgery. This decrement of escape threshold lasted for more than 3 weeks. The whisker pad area contralateral to the IAN transection also showed a decrease in escape threshold to non-noxious mechanical stimulation as compared with sham-operated rats. However, the change in threshold intensity for the side contralateral to transection was not as pronounced as that on the ipsilateral side. Fos protein-like immunoreactive (LI) cells were observed in the superficial laminae but not dominant in deeper laminae of the trigeminal spinal nucleus caudalis (Vc) and the first segment of the spinal cord (C1) after non-noxious mechanical stimulation of the whisker pad area in the rats with IAN transection. Fos protein-LI cells were expressed bilaterally in the Vc and C1, but were more numerous on the ipsilateral side to transection than on the contralateral side. The largest number of Fos protein-LI cells was observed at 2400 microm caudal from the trigeminal subnucleus interporalis (Vi)-Vc border both in ipsilateral and contralateral sides. The number of Fos protein-LI cells increased after application of 1, 4, and 16 g stimuli as compared to rats without mechanical stimulation. Furthermore, an extensively greater number of Fos protein-LI cells were expressed both in superficial and deep laminae of the bilateral Vc and C1 of the spinal cord after subcutaneous injection of mustard oil into the whisker pad. Fos protein expression after mustard oil injection was much stronger than that observed after any mechanical stimulation in the rats with IAN transection. These data suggest that the change in the numbers and spatial arrangement of nociceptive neurons in the Vc and C1 after IAN transection reflect the development of mechanical hyperalgesia in the area adjacent to the IAN innervated region.     

次声导致小鼠脑损伤的生化指标研究

目的研究次声影响小鼠脑组织的生化指标。方法BALB/C小鼠暴露于16Hz、声压90dB次声,2h/d,分别作用1、7、14、21和28d后,采用免疫组织化学方法观察小鼠脑中胶质原纤维酸性蛋白(GFAP)的表达。结果次声作用一定时间后,GFAP阳性星形细胞主要分布在海马、皮质、下丘脑等脑区,且随次声作用天数增加,GFAP阳性星形细胞数目增多。结论GFAF阳性细胞的增加可以证实次声对以上这些脑区有不同程度的损伤。     

The effects of ibuprofen and the neurokinin‐1 receptor antagonist GR205171A on Fos expression in the ferret trigeminal nucleus following tooth pulp stimulation

We have developed a model to study central changes following inflammation of the tooth pulp in the ferret and have examined Fos expression in the trigeminal nucleus following stimulation of non‐inflamed and inflamed tooth pulps. The aim of this study was to establish the ability of this model to predict analgesic efficacy in clinical studies of inflammatory pain. We addressed this by assessing the effects of the neurokinin‐1 receptor antagonist GR205171A and ibuprofen on Fos expression following stimulation of the inflamed pulp and comparing this with known analgesic efficacy. Adult ferrets were prepared under anaesthesia to allow tooth pulp stimulation, recording from the digastric muscle and intravenous injections at a subsequent experiment. In some animals pulpal inflammation was induced, by introducing human caries into a deep buccal cavity. After 5 days, animals were reanaesthetised, treated with vehicle, GR205171A or ibuprofen and the teeth were stimulated at ten times the threshold of the jaw‐opening reflex. Stimulation of all tooth pulps induced ipsilateral Fos in trigeminal subnuclei caudalis and oralis. GR205171A had no significant effect on Fos expression in the trigeminal nucleus of animals with either non‐inflamed or inflamed tooth pulps. Ibuprofen reduced Fos expression in the trigeminal nucleus and this effect was most marked in animals with pulpal inflammation. These results differ from those previously described using a range of other animal models, but agree with known clinical efficacy of neurokinin‐1 receptor antagonists and ibuprofen. Therefore this model is likely to be of use in accurately predicting the analgesic efficacy of novel compounds.