正交试验法优选七君颗粒提取工艺

目的:以三七皂苷R1、人参皂苷Rg1的含量为指标,研究七君颗粒中三七、人参等10味中药的提取工艺。方法:采用正交试验设计法,对七君颗粒水提取工艺进行考察,用高效液相色谱法测定三七皂苷R1、人参皂苷Rg1含量。结果:加水量12倍,煎煮1次,煎煮时间每次90min,所得提取液中三七皂苷R1、人参皂苷Rg1的含量高。结论:此工艺对三七皂苷R1、人参皂苷Rg1的含量的提取率高,稳定性好,可作为七君颗粒的提取工艺。     

Anisomycin uses multiple mechanisms to stimulate mitogen-activated protein kinases and gene expression and to inhibit neuronal differentiation in PC12 phaeochromocytoma cells

Treatment of PC12 cells with nerve growth factor (NGF) stimulates extracellular signal-regulated kinases (ERKs), as well as stress-activated c-Jun N-terminal kinases (JNKs) and p38 kinase, and induces neuronal differentiation. While the pivotal role of ERKs in NGF-induced morphological differentiation is well established, the contribution of JNK- and p38-pathways is less clear. The role of the JNK- and p38-pathway in PC12 cells was analysed by using anisomycin, a protein synthesis inhibitor that activates JNKs and p38. Non-toxic concentrations of anisomycin were found to stimulate these enzyme activities as well as the expression of the early response genes c-jun, c-fos and zif268, and to inhibit NGF-induced neurite formation. These effects of anisomycin appear to be mediated by the generation of reactive oxygen species (ROS), which in turn act through both TrkA/Ras-dependent and -independent signalling pathways. In addition, cross-talk between the p38- and ERK-pathways appears to play a role in the action of anisomycin.     

慎用大承气汤治疗机械性肠梗阻

大承气汤出自《伤寒杂病论》,是张仲景为治疗阳明腑实证而立,在临床上大承气汤应用广泛。许多学者将大承气汤作为治疗肠梗阻的主方。张仲景在大承气汤证中言腹满痛,腹满而喘,腹中转矢气等;在小承气汤证中言“腹大满不通者,可与小承气汤”。以此为界定标准。这些说明了张仲景使用大承汤是非常慎重的。并非肠腑不通就用大承气汤。肠梗阻有虚、实之分,缓、急之别,程度之异,临床上机械性肠梗阻使用大承气汤疗效也不理想。因此在治疗上必须详审病因,知犯何逆,随证治之,不可一味攻之。     

Gene expression profiling of drug metabolism and toxicology markers using a low-density DNA microarray

DNA microarrays are useful tools to study changes of gene expression in response to a treatment with drugs. Here, we describe the optimization of conditions for the cDNA synthesis and hybridization protocols to be used for a low-density DNA microarray called 'Rat HepatoChips.' This DNA microarray with 59 carefully selected genes could be used to study changes in gene expression levels due to a treatment with xenobiotic. These 59 genes (including 8 housekeeping genes) have been selected among potential toxic markers involved in basic cellular processes and drug metabolism related genes. Using the optimized conditions, the results were shown to be reproducible, with 6% variation between the duplicated spots and 10% between arrays. Conditions were optimized to allow quantification with a dynamic range of four log units. In order to demonstrate the major advantage of these tool for studying gene expression, samples of control rat liver were compared with those of animals dosed with phenobarbital (PB) or pregnenolone-16 alpha-carbonitrile (PCN), two compounds well known to induce cytochrome P450 isoforms of 2B and 3A subfamilies, respectively. This microarray has shown that other genes apart from the corresponding CYP P450 genes have been changed due to PB and PCN treatment. Apoptosis-related genes have shown to be changed due to PB and PCN treatment, which confirms results from previous work.     

脑溢安对出血性中风大鼠脑内磷酸化蛋白激酶表达的影响

探讨脑溢安对出血性中风大鼠脑内磷酸化蛋白激酶（P－C－Jun）表达的影响和治疗机理。根据 Roserberg法复制大鼠脑出血模型,采用免疫组织化学方法观察脑出血后梨状皮质内P－C－Jun的表达变化,并用组织病理学方法分析脑皮质内神经元的损伤状况。结果：脑溢安治疗组较模型组脑内P－C－Jun蛋白表达弱,神经元受损状况轻。结论：脑溢安下调脑内P－C－Jun蛋白表达,从而干预了丝裂原活化的蛋白激酶（MAPK）信号转导通路。     

低氧预处理对缺氧-复氧后体外培养大鼠海马神经元Fos、Jun表达和神经元凋亡的影响

目的　观察低氧预处理对体外培养大鼠海马神经元缺氧 复氧后Fos、Jun表达和神经元凋亡的变化。方法　取培养 12d的两组 (对照组和低氧预处理组 )神经元 ,同时置于缺氧环境 (0 90l LN2 、0 10l LCO2 )中培养 4h后取出 ,置含 0 10l LCO2 和空气的培养箱内复氧培养 2 4和 72h。于不同时间取出 ,观察神经元存活数 ,并分别用抗Fos和Jun抗血清进行免疫组织化学染色 ,观察Fos、Jun表达阳性和阴性神经元数目 ,计数Fos和Jun表达神经元所占百分率。并用原位末端标记 (TUNEL)法和流式细胞术分别检测缺氧 复氧对体外培养海马神经元凋亡的影响。　结果　缺氧 复氧后大鼠海马培养神经元中Fos、Jun表达阳性神经元百分率和凋亡神经元百分率均较缺氧前显著增加。经低氧预处理的海马神经元缺氧 复氧后Fos、Jun表达神经元和凋亡神经元百分率均较对照组明显减少。神经元损伤程度减轻 ,神经元存活数明显高于对照组。　结论　低氧预处理可使培养中海马神经元对缺氧产生耐受 ,减少缺氧 复氧后神经元Fos和Jun表达 ,减少缺氧 复氧后神经元凋亡。     

Mechanism of Fibroblast‐Like Synoviocyte Apoptosis Induced by Recombinant Human Endostatin in Rats with Adjuvant Arthritis

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by pronounced synovial hyperplasia, in which there may be an imbalance between the growth and death of fibroblast‐like synoviocytes (FLS). The present study was undertaken to examine the effect of recombinant human endostatin (rhEndostatin) on FLS apoptosis in experimental RA. Adjuvant arthritis (AA) was induced in male Sprague Dawley (SD) rats. Using cultured AA FLS obtained from these rats, the apoptosis process was measured by terminal deoxyribonucleotidyl transferase‐mediated dUTP nick‐end labeling (TUNEL) as well as Annexin V‐fluorescein isothiocyanate (FITC) and propidium iodide (PI) labeling methods. In addition, the expression levels of the Fas, c‐jun, NFκB, and caspase‐3 gene products in synovial tissues were quantified by quantitative real‐time polymerase chain reaction (qPCR) and/or Western blotting assays. Our data revealed that rhEndostatin induced apoptosis in AA FLS. The number and signal density of TUNEL‐positive cells were significantly increased in rats treated with rhEndostatin (2.5 mg/kg). The percentage of Annexin V‐FITC‐positive cells was 6.67% after treatment with rhEndostatin at 25 μg/mL for 48 hr, compared with only 3.32% among untreated control cells. There were significant increases in Fas mRNA, c‐jun mRNA, c‐Jun protein, and caspase‐3 (p20) protein in AA synovial tissues treated with rhEndostatin (2.5 mg/kg), whereas no significant difference in NFκB expression was detected between treated and untreated tissues. These findings indicate that rhEndostatin has a therapeutic effect on RA by inducing FLS apoptosis, which is strongly associated with increased expression of Fas, c‐jun, and caspase‐3, but not NFκB. Anat Rec, 291:1029–1037, 2008. © 2008 Wiley‐Liss, Inc.     

Dexmedetomidine‐induced contraction involves c‐Jun NH2‐terminal kinase phosphorylation through activation of the 5‐lipoxygenase pathway in the isolated endothelium‐denuded rat aorta

Vasoconstriction induced by dexmedetomidine, a highly selective alpha‐2 adrenoceptor agonist, mainly involves c‐Jun NH₂‐terminal kinase (JNK) phosphorylation in the isolated endothelium‐denuded aorta. We carried out an in vitro study to determine the main arachidonic acid metabolic pathway that is involved in dexmedetomidine‐induced JNK activation. Cumulative dexmedetomidine concentration‐contractile response curves were generated in the endothelium‐denuded rat aorta in the presence or absence of the following inhibitors: the JNK inhibitor SP600125, the phospholipase A₂ inhibitor quinacrine dihydrochloride, the non‐specific lipoxygenase (LOX) inhibitor nordihydroguaiaretic acid, the 5‐LOX inhibitor AA‐861, the dual 5‐LOX and cyclooxygenase (COX) inhibitor phenidone, the non‐specific COX inhibitor indomethacin, the cytochrome p450 epoxygenase inhibitor fluconazole, the COX‐1 inhibitor SC‐560, and the COX‐2 inhibitor NS‐398. The effect of the alpha‐2 adrenoceptor inhibitor rauwolscine and other inhibitors, such as quinacrine dihydrochloride, nordihydroguaiaretic acid, AA‐861, phenidone, indomethacin and the protein kinase C inhibitor GF 109203X, on dexmedetomidine‐induced JNK phosphorylation was investigated in rat aortic vascular smooth muscle cells with western blotting. The effect of dexmedetomidine on 5‐LOX and COX‐2 expression was investigated in vascular smooth muscle cells. SP600125, quinacrine dihydrochloride, nordihydroguaiaretic acid, AA‐861, phenidone, rauwolscine and chelerythrine attenuated dexmedetomidine‐induced contraction. Indomethacin slightly attenuated dexmedetomidine‐induced contraction. Fluconazole and SC‐560 had no effect on dexmedetomidine‐induced contraction, whereas NS‐398 attenuated contraction. SP600125, rauwolscine, quinacrine dihydrochloride, nordihydroguaiaretic acid, AA‐861, phenidone and GF 109203X attenuated dexmedetomidine‐induced JNK phosphorylation. 5‐LOX and COX‐2 were upregulated by dexmedetomidine. Thus, dexmedetomidine‐induced alpha‐2 adrenoceptor‐mediated contraction is mediated mainly by 5‐LOX and partially by COX‐2, which leads to JNK phosphorylation.     

杨巍运用培源滑润法治疗便秘经验采撷

[目的]总结舒鹏教授运用健脾化瘀法治疗胃癌的相关学术思想及临床经验。[方法]通过跟随舒鹏教授临证抄方及对其治疗胃癌典型病案的收集、整理,从胃癌的病机特点、遣方用药等方面阐述舒鹏教授健脾化瘀法论治胃癌的相关经验,并附验案一则加以佐证。[结果]胃癌的病机以脾胃虚弱为本,病理特点以瘀毒为患,脾虚瘀毒是胃癌的重要病机。在胃癌的治疗上,强调以健脾养胃扶正为本,临证时病症合参,根据胃癌不同分期的临床特点,分期论治,以求精准治疗,并针对胃癌"瘀毒"的病理特点,适当运用活血化瘀及虫类药物化瘀解毒。验案中患者为胃癌I期术后,以六君子汤为基础方健脾养胃以扶正,辅以活血化瘀解毒之品以消潜在之瘀毒,以防复发、转移,显著改善患者的临床症状,治疗至今,期间定期复查,病情稳定。[结论]舒鹏教授以健脾扶正、化瘀解毒法分期论治胃癌,临床疗效颇佳,值得学习、研究与探讨。