颈性眩晕的针灸与养血清脑颗粒治疗

目的 探讨针灸与养血清脑颗粒治疗颈性眩晕的临床疗效。方法 选择诊断明确的颈性眩晕患者病例，随机分为两组，治疗组予以针灸治疗结束后口服养血清脑颗粒。对照组仅采用口服养血清脑颗粒。结果治疗组经颅多普勒超声检查椎基底动脉血流改善明显优于对照组（P〈0．05），治疗组自觉症状消失时间明显比对照组时间缩短（P〈0．01），随访半年、复发率明显优于对照组（P〈0．01）。结论 针灸与养血清脑颗粒治疗颈性眩晕可以明显提高疗效。     

眼络通颗粒的提取工艺研究

目的:研究眼络通颗粒的提取工艺。方法:采用正交设计试验,以葛根中葛根素转移率为指标,考察乙醇浓度、溶剂倍数、提取时间、提取次数对葛根乙醇提取效率的影响;采用水蒸气蒸馏法提取川芎、三棱中的挥发油,并制成β-环糊精包合物;采用水煎煮法提取其它药材中的水溶性成分。结果:葛根的最佳提取条件为取葛根药材8倍量的70%乙醇浸泡75min,再加热回流提取3次,每次2h;挥发油β-环糊精包合物制备方法为取15倍处方量川芎、三棱药材,加10倍量水,水蒸气蒸馏提取4h,用β-环糊精15g进行包合。结论:本工艺条件能最大限度提取药材中的有效成分。     

肝宁颗粒的质量标准研究

目的:建立肝宁颗粒的质量标准。方法:采用薄层色谱法对方中君药川芎进行定性鉴别,以高效液相色谱法测定川芎中阿魏酸的含量。结果:供试品色谱中,在与川芎对照药材相应的位置上,显相同颜色的荧光斑点;阿魏酸进样量在0·0384μg~0·2304μg范围内与峰面积积分值线性关系良好(r=0·9999),平均加样回收率为100·1%(RSD=0·91%)。结论:建立的质量标准可有效控制肝宁颗粒的质量。     

宫宁颗粒质量标准研究

目的:建立宫宁颗粒的质量标准。方法:采用薄层色谱法对宫宁颗粒中的黄芩苷进行定性鉴别;以高效液相色谱法测定黄芩苷的含量。结果:供试品薄层色谱中,在与黄芩苷对照品相应的位置上显相同颜色的斑点;黄芩苷检测浓度在20～100μg/ml范围内与峰面积积分值线性关系良好,平均回收率为99.71%(RSD=1.23%)。结论:所建质量标准可用于本品的质量控制。     

Selective and AMPA receptor-dependent astrocyte death following prolonged blockade of glutamate reuptake in rat cerebellar cultures

In this study we examined the effects of prolonged l-trans-pyrrolidine-2,4-dicarboxylate (PDC)-induced glutamate reuptake blockade on the viability of glial cells in cerebellar granule cell cultures. Immunofluorescence staining for the glial-specific intermediate filament protein, GFAP, revealed that the PDC- induced increase of extracellular glutamate concentration was accompanied by increased astrocyte death, while neurons and oligodendrocytes remained intact and viable. Astrocytic cell death was manifested as fragmentation of processes and cell bodies. The selective astrocyte death was completely prevented by the competitive alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate receptor antagonist, NBQX (10 microM), whereas MK-801 (10 microM), a noncompetitive blocker of N-methyl-D-aspartate receptors, gave only partial protection. Double staining for GFAP and the AMPA receptor subunits GluR2/3 showed that astrocytes had much higher immunoreactivity for GluR2/3 than neurons or oligodendrocytes, suggesting that the number of AMPA receptors is likely to be higher on astrocytes. Furthermore, we employed real-time RT-PCR to measure GluR1-4 subunit mRNA expression in control and PDC-exposed cultures. Following treatment with PDC, GluR1 and GluR4 mRNAs were reduced by 40% and GluR3 was reduced by 70% relative to control levels. In contrast, GluR2 expression was not affected by the PDC treatment, indicating that GluR3 was the dominant type of AMPA receptor subunit expressed on astrocytes. Our results show that astrocytes appear to be more vulnerable than neurons or oligodendrocytes to a gradual increase in the extracellular glutamate concentration, suggesting that astrocytes may be critically involved in the pathophysiology of slowly developing chronic neurodegenerative disorders.     

Different dependence of lithium and valproate on PI3K/PKB pathway

Objectives: Acute treatment with valproate (VPA) or lithium (Li⁺) protects cerebellar granule cells (CGC) against apoptosis induced by low potassium (K⁺) (5 mM). As the protection induced by VPA is absolutely dependent on insulin, in contrast to the observed effects of Li⁺, we decided to study the different role of the PI3K/PKB pathway in the neuroprotective effects of both drugs.

Methods: We have studied the neuroprotection elicited by Li⁺ or VPA in cultures of rat CGC. We induced the apoptosis by switching to a medium with a low concentration of K⁺ or by adding C₂-ceramide to the cultures. We studied the effect of Li⁺ and VPA on viability and on the regulation of the PI3K/PKB pathway.

Results and conclusions: Insulin also protects against low K⁺-induced apoptosis in CGC, probably through its interaction with an insulin-like growth factor receptor. Moreover, whereas Li⁺ protects against the apoptosis induced by C₂-ceramide, VPA cannot, probably due to the inhibition of protein kinase B (PKB) caused in this apoptotic stimulus. These results suggest that VPA protects against low K⁺-induced apoptosis by acting on the PI3K/PKB pathway; however, VPA does not affect the increase of PKB activity caused by insulin in these cells. The protection by Li⁺ is independent of this transduction pathway. Moreover, Li⁺ blocks the caspase 3 activation induced by low K⁺, whereas neither VPA nor insulin affects this activation.     

The total numbers of cerebellar granule neurons in young and aged Fischer 344 and Wistar-Kyoto rats do not change as a result of lengthy ethanol treatment

It is generally accepted that long term chronic ethanol consumption by young rats will lead to significant losses of cerebellar granule neurons (GN). A recent study in this laboratory showed, however, that 40 weeks of chronic ethanol consumption had no effect on the total numbers of GN in aged Fischer 344 rats (F344). The goals of the present study were to determine whether F344 GN were resistant to ethanol toxicity only in aged rats and whether resistance of GN in aged rats to ethanol toxicity occurred only in the F344 strain. To accomplish those goals, young and aged adult F344 and Wistar-Kyoto (WKY) rats were treated chronically with ethanol for 40 weeks during the first or second half of their life span. In each rat the total numbers of GN were estimated with the optical fractionator and the volumes of the GN layer were estimated according to Cavalieri's theorem. After the 40 weeks of ethanol, there were significant age-related differences in the total numbers of GN in the F344 rats. There were also significant strain-related differences in the total numbers of GN and volumes of the GN layer. There were no significant ethanol-related differences, however, in numbers of cerebellar GN or volumes of the GN layer in F344 rats or WKY rats. The results presented here show that consumption of ethanol over long periods of time had no effect on the total numbers of cerebellar GN or the granular layer volumes in young or aged F344 or WKY rats.     

扶正抑瘤颗粒对乳腺癌核转录因子-kB及细胞周期的影响

目的:研究扶正抑瘤颗粒(FYK)对乳腺癌肿瘤组织核转录因子-κB(NF-κB)和细胞周期的影响及其临床意义。方法:55例乳腺癌患者,随机分为治疗组和对照组,两组均用常规化疗,治疗组同时口服FYK,1个月后取肿瘤组织用流式细胞仪检测NF-κB的表达情况及肿瘤细胞周期各时相的比例。结果:治疗组与对照组比较,NF-κB的表达明显提高(P<0.01);G_(0/1)期细胞比例明显升高(P<0.01),S期细胞比例及细胞增殖指数(PI)明显下降(P<0.01)。结论:FYK可提高乳腺癌组织NF-κB表达水平;能升高G_(0/1)期细胞比例,降低S期细胞比例及PI值,对乳腺癌患者的预后起积极作用。     

Protoporphyrin IX production and its photodynamic effects on glioma cells, neuroblastoma cells and normal cerebellar granule cells in vitro with 5-aminolevulinic acid and its hexylester

5-Aminolevulinic acid (ALA) has shown promising in photodynamic detection and therapy of brain tumor. However, the knowledge on selective accumulation of ALA-induced protoporphyrin IX (PpIX) in brain tumor tissue is still fragment. In the present study, the rat C6 glioma cells, human SK-N-SH neuroblastoma cells, and rat normal cerebellar granule cells (RCG) were used to investigate the PpIX production and photocytotoxicity in vitro. The C6 cells and SK-N-SH cells showed a similar kinetics of PpIX accumulation after exposure to ALA or ALA hexyl ester (ALA-H), with an initial increase up to 6–8 h and then saturated. In the case of RCG cells, the PpIX accumulation slowly increased until 12 h studied. However the cellular PpIX content was more than 10 times higher in the C6 and SK-N-SH cells than that in the normal RCG cells. The intracellular localization of PpIX measured by cofocal laser scanning microscopy was in same pattern in the C6 glioma cells and RCG normal cells with a diffuse cytoplasm distribution. The sensitivity of the C6 cells and SK-N-SH cells to ALA or ALA-H PDT was similar. It appears that ALA-H could achieve similar or slightly better results than ALA with respect to PpIX production and photoinactivation of cells, although a 10 times lower concentration of ALA-H was used.