姜黄素对哮喘大鼠肺内胶原沉积、结缔组织生长因子表达及气道重构的影响

目的:通过观察姜黄素对哮喘大鼠肺内胶原沉积、结缔组织生长因子(connective tissue growth factor,CTGF)表达的影响,探讨姜黄素对哮喘气道重构的治疗作用。方法:36只雄性SD大鼠随机分为正常对照组(A组)、哮喘模型组(B组)、姜黄素治疗组(C组)各12只,采用卵蛋白致敏大鼠哮喘模型,观察3组动物哮喘发作症状、胶原沉积情况及SABC免疫组化法检测CTGF在肺组织的表达变化。结果:B组大鼠哮喘发作症状明显重于C组,A组无症状。B组支气管黏膜下、血管壁及周围、肺泡壁、肺间质均有胶原大量沉积,C组胶原沉积程度明显轻于B组。A组胶原沉积不明显。免疫组化发现B组CTGF表达最强,C组较弱,A组微弱表达。结论:姜黄素可抑制哮喘大鼠肺组织的胶原沉积,在延缓气道重建中起重要作用,其作用机制可能是通过抑制CTGF的表达来实现。     

Evidence for A1 and A3 receptors mediating adenosine-induced intracellular calcium release in the dorsal root ganglion neurons by using confocal microscopy imaging

Adenosine exerts a key role in analgesia. In the present study, adenosine-induced Ca²⁺ responses were revealed by using confocal microscopy imaging in the rat dorsal root ganglia (DRG) neurons in vitro. Our results showed that adenosine could evoke increases in the intracellular Ca²⁺ concentration in the DRG neurons. In addition, by application of selective receptor antagonists, two types of receptors, A₁R and A₃R, were identified to be involved in the adenosine-induced Ca²⁺ release from intracellular stores in neurons. Altogether, these results suggest that confocal microscopy imaging combined with fluorescent dyes could help to detect the analgesic-induced ion signaling in single cell.     

A retrospective analysis of pathological changes of testicular tissue in normal adult rats

Rat testicular model is widely used in experiments on andrology, pharmacology and reproductive toxicology. Generally, normal adult rat is considered to have normal testes. However, whether normal adult rats appeared abnormal testes have not been evaluated. The objective of this study was to evaluate the incidence of abnormal testes in normal adult Sprague Dawley (SD) rats and pathological changes in testicular tissues. Six hundred and sixteen adult male SD rats used in previous studies as controls were retrospectively analysed. Testicular tissues were stained with haematoxylin–eosin for observation of pathology. Among 616 rats, 14 rats had pathological testes, and the incidence of abnormal testis was 2.3%. In the 14 rats with abnormal testes, 10 rats were microrchidia (71.4%) and four rats showed normal testicular size. Testicular abnormality included complete interruption of spermatogenesis, partial germ cell arrest, progressive hypospermatogenesis, seminiferous epithelia vacuolation and inflammatory status. Bilateral testicular tissues had similar pathological changes in abnormal testes. The findings in this study demonstrate that the normal adult rats have abnormal testes. We should pay attention to the possibility of abnormal testes when using normal adult male rats for establishing a testicular model.     

shRNA-mediated GSTP1 gene silencing enhances androgen-independent cell line DU145 chemosensitivity

Purpose

Design short hairpin RNA (shRNA) interference sequence to silence glutathione S-transferase P1 (GSTP1) gene of androgen-independent prostate cancer cell line DU145, and then to explore its effect on sensitivity to chemotherapeutics.

Methods

Target sequence was picked up to form the shRNA. DU145 cell was divided into five groups according to the shRNA added for transfection: shRNA255, shRNA554, shRNA593, negative-shRNA and blank group. Fluorescence microscope was used to pick up the shRNA with the highest transfection ratio. Western blotting and RT-PCR were taken to pick up the shRNA with the best gene silencing result. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay and terminal de-oxynucleotidyl transferase-mediated dUTP nick end-labeling assay were used to detect survival ratio and apoptosis ratio of DU145 administered of fluorouracil (5-FU) or paclitaxel (PA) at different concentrations before and after shRNA transfection.

Results

Three different shRNA oligonucleotides (shRNA255; shRNA554; shRNA593) targeting the coding sequence of GSTP1 mRNA and one negative control shRNA were constructed. The transfection ratio of shRNA554 (76.2 ± 0.68 %) was higher than that of shRNA255 (63.3 ± 1.04 %) (P < 0.01) or shRNA593 (72.7 ± 0.33 %) (P < 0.01). After transfection of shRNA554, the mRNA and protein of level were the lowest, P < 0.01. The survival ratio of DU145 administered with 5-FU of different concentrations (30, 60, 120, 240 μg/ml) declined after transfection (P < 0.01). Besides, the apoptosis ratio increased after transfection (P < 0.01). Similarly the survival ratio of DU145 administered with PA of different concentrations (0.2, 2, 10, 20 μg/ml) declined (P < 0.01) and the apoptosis ratio increased (P < 0.01) after transfection.

Conclusions

The gene GSTP1 silence via shRNA transfection to androgen-independent prostate cancer cell line DU145 enhances the sensitivity to chemotherapeutics.     

癫痫与心脏自主神经功能障碍

目的：分析癫痫患者发作期和发作间期自主神经功能障碍与癫痫患者突然意外死亡的关系。 资料来源：应用计算机检索Medline数据库1985-01／2004-12期间的相关文章限定文章语言种类为英文，检索词”epilepsy and autonomic”和“seizure and autonomic”。 资料选择：对资料进行初审，纳入标准：①癫痫和癫痫发作与自主神经功能状态关系。②癫痫突然意外死亡。 资料提炼：共收集到符合上述要求的文献45篇，排除27篇重复性内容。18篇符合纳入标准：其中16篇关于癫痫和癫痫发作与自主神经功能障碍，2篇关于自主神经功能障碍与癫痫患者突然意外死亡的关系。 资料综合：癫痫和癫痫发作均可引起心脏自主神经功能障碍，无论部分性发作还是全身性发作癫痫患者发作期和发作间期都存在自主神经功能障碍。而心脏自主神经功能障碍与心律失常尤其是恶性心律失常的发生密切相关。 结论：癫痫患者发作期和发作间期心脏自主神经功能障碍引发的恶性心率失常可能是癫痫患者发生突然意外死亡的重要原因。