桃仁-红花"药对"不同配比的血府逐瘀汤对急性心肌缺血的保护作用

目的观察桃仁-红花“药对”不同配比的血府逐瘀汤抗垂体后叶素(Pit)致大鼠急性心肌缺血的作用。方法采用大鼠舌下静脉注射Pit造成急性心肌缺血模型,观察各给药组注射Pit后不同时间点Ⅱ导联心电图ST段变化以及血清乳酸脱氢酶(LDH)、肌酸激酶(CK)、超氧化物歧化酶(SOD)、丙二醛(MDA)的含量。结果桃仁、红花不同配比的血府逐瘀汤对Pit所致缺血性ST段有明显改善作用,能明显增加血清SOD的活性,降低MDA、CK和LDH的含量,其中桃仁、红花配比为4∶3时效果最为显著。结论桃仁、红花不同配比的血府逐瘀汤均具有明显对抗Pit致大鼠急性心肌缺血的作用,其功效与桃仁-红花的不同配比之间关系不明显。     

肺癌患者外周血CK19mRNA、CEA、NSE、TPA联合检测的意义

背景及目的:细胞角蛋白作为检测循环癌细胞的指标,越来越得到人们的重视。CK19mRNA作为微转移检测的基因标记有较高的应用价值,但其特异性及敏感度都不够理想。本实验旨在评价外周血CK19mRNA与肿瘤标记物癌胚抗原(CEA)、神经元特异性烯醇酶(NSE)和组织多肽抗原(TPA)水平的联合测定对肺癌临床诊断的价值。方法:用逆转录多聚酶链反应法(RT-PCR)检测外周血有核细胞CK19mRNA的表达水平,采用酶联免疫吸附法(ELISA)和时间分辨荧光法(TRF)测定91例肺癌、33例良性肺病患者和50例健康人的血清CEA、NSE和TPA水平,数据统计用t检验和χ2检验分析。结果:肺癌组外周血CK19mRNA和血清CEA、NSE、TPA的阳性检出率(分别为49.5%、65.9%、67.0%、83.5%)明显高于良性肺病组(分别为24.2%、9.0%、15.2%、9.0%)和健康对照组(分别为10.0%、4.0%、8.0%、6.0%)(P<0.01)。在不同的组织类型中CK19mRNA阳性检出率无统计学差异。随着临床分期逐步升高,Ⅰ+Ⅱ期、Ⅲ期、Ⅳ期CK19mRNA的阳性检出率分别为44.4%、48.4%、54.2%呈上升趋势,但无显著性差异(P>0.05)。在Ⅰ+Ⅱ期、Ⅲ期、Ⅳ期血清CEA水平犤(μg/L)/阳性率(%)犦分别为3.5/27.8、5.1/35.5、8.5/58.3,血清NSE水平犤(μg/L)/阳性率(%)犦分别为12.5/33.3、15.3/41.9、24.7/58.3,血清TPA     

35例高活性CK标本的CK-MB质量及活性检濒结果分析

目的探讨对高活性CK标本进行CK—MB质量及活性检测比较,为临床提供准确结果。方法将35例高活性cK标本分别在全自动化学发光仪及全自动生化分析仪上检测其CK—MB质量及活性。结果CK—MB活性的测定结果与实际值相比远远要高;CK—MB质量检测可有效减少检测干扰,有效保证可靠的检测结果。结论高活性CK标本严重影响其CK—MB活性检测,但对CK—MB质量检测影响不大。     

甲状腺乳头状癌与乳头状增生的病理研究

  目的 探讨Galectin3，CK19及Ki-67在甲状腺乳头状癌与乳头状增生中的表达，寻找有助于两者鉴别诊断的标志物。方法 运用免疫组化方法检测100例甲状腺乳头状癌、100例良性乳头状增生中Galectin3，CK19及Ki-67的表达。结果 Galectin3，CK19及Ki-67在甲状腺乳头状癌阳性表达率分别为100 %，97 %及93 %，而在乳头状增生中表达率分别为13 %，30 %及1 %，3种蛋白在乳头状癌与良性乳头状增生间差异有统计学意义（P＜0.05）。在乳头状癌中2种或3种蛋白同时阳性表达为94.3 %，而乳头状增生为0。结论 Galectin3，CK19及Ki-67是鉴别诊断甲状腺乳头状癌与乳头状增生的有用标志物，尤其联合使用更有价值。     

体外培养不同代角膜缘干细胞特性分析

目的比较不同代角膜缘干细胞的生物学特性,为角膜缘细胞移植在临床中的氉应用提供依据。方法组织块培养法体外培养角膜缘干细胞,光镜观察、HE染色和免疫荧光染色分析原代、第1代和第2代角膜缘细胞形态学特性;流式细胞技术定量检测原代、第1代和第2代角膜缘细胞ABCG2表达;MTT比色法测定体外培养原代、第1代和第2代角膜缘干细胞的增殖活性变化。结果倒置相差显微镜下可见原代细胞培养24h有单个细胞从组织块边缘迁出,48h细胞局部可见拉网样生长趋势,72h出现大片细胞,细胞呈现多边形或多角形,96h细胞几乎铺满整个培养板。随着传代次数的增加,细胞形态变得不规则,CK3单克隆抗体表达阳性率逐渐增强;p63单克隆抗体原代和第1代细胞间表达阳性率无明显差别,第2代细胞表达量明显减弱。流式细胞检测显示原代、第1代和第2代细胞ABCG2表达阳性率分别为13.41%、22.96%和4.43%。MTT结果显示随传代次数增加细胞增殖活性逐渐下降。结论原代和第1代角膜缘干细胞可以作为制备组织工程角膜上皮细胞膜片的较理想种子细胞。     

细胞角蛋白7、20在女性腺性膀胱炎中的表达及临床意义

目的探讨女性腺性膀胱炎中细胞角蛋白7、20(CK7、CK20)的表达及临床意义。方法对20例女性膀胱移行细胞癌(TCC)组织、30例女性腺性膀胱炎组织、20例正常膀胱移行上皮组织采用免疫组织化学Envision二步法进行检测。结果 CK7的表达在女性腺性膀胱炎与膀胱癌之间有统计学差异(χ~2=5.36;P<0.05),与正常膀胱组织之间无统计学差异(χ~2=1.09;P>0.10),但是其在腺性膀胱炎中表达率(60.00%)较正常膀胱组织(45.00%)高。CK20的表达在腺性膀胱炎和膀胱癌之间有统计学差异(χ~2=7.41;P<0.01),与正常膀胱组织之间无统计学差异(χ~2=1.63;P>0.10);腺性膀胱炎的CK20表达率(53.33%)明显高于正常膀胱组织(35.00%)。结论女性腺性膀胱炎转变为膀胱移行细胞癌倾向性较低,是具有一定恶变趋向的良性病变。     

Acute and subchronic oral toxicity assessment of the herbal formula Kai-Xin-San

Ethnopharmacological relevance

Kai-Xin-San (KXS) is a famous traditional Chinese medicine (TCM) formula. It has been used in the treatment of diseases including neurasthenia, Alzheimer's disease and neurosis.

Aim of the study

To provide information on the potential toxicity of KXS, we evaluated the acute and subchronic toxicity in rodents.

Materials and methods

In acute study, a single administration of KXS was given orally to mice at doses ranging from 19.67 to 60.04 g/kg. In the sub-chronic oral toxicity study, KXS was administered to rats at 0, 1, 3 and 9 g/kg for 13 weeks. Moreover, 30 days of post treatment (withdrawal study) was conducted. Mortalities, clinical signs, body weight changes, food and water consumption, haematological and biochemical parameters, gross findings and organ weights were monitored during the study period.

Results

In the sub-chronic study in rats, daily oral administration of KXS at the dose of 9 g/kg/day result in significant increase in WBC, lymphocyte, alkaline phosphatase, blood sugar and significant decrease in bodyweight, serum Cre, CK and CHO at the last week of treatment. Recovery except for the body weight was observed after 30 days of post treatment.

Conclusions

KXS is relatively safe for oral medication. The LD₅₀ of KXS was over 32.59 g/kg for mice. The no-observed-adverse-effect-level (NOAEL) was considered to be 19.67 g/kg/day for rats.     

降钙素基因相关肽参与电针抗急性心肌缺血损伤的实验研究

目的:探讨电针刺激抗心肌缺血的机制。方法:采用异丙肾上腺素(Isoprenolonhydrochloride,ISO)腹腔注射建立大鼠急性心肌缺血实验模型。24只雄性Wistar大鼠随机分为3组,每组8只。①电针治疗组:将大鼠麻醉后,注射2%ISO,按100mg/kg(即5mL/kg),i.p.,同时进行电针治疗。选取左侧“心俞”接正极和“厥阴俞”接负极。电针参数:输出波形为正负向交替脉冲波,脉冲宽度为300μs,频率2-20Hz,疏密波,输出电压为3.5-5V,以针柄轻度抖动为度,每次电针30min,共2次,间隔12h。②模型组:同电针治疗组,但不进行电针治疗。③对照组:将大鼠麻醉后,注射生理盐水,38℃,按5mL/kg体重i.p.。比色法测定血清肌酸激酶(creatinekinase,CK)值;放射免疫法测定血浆、心脏组织、脊髓(T1-T6)组织内降钙素基因相关肽(CGRP)含量。结果:电针治疗能够降低造模引起的血清CK水平升高;急性心肌缺血损伤可导致血浆CGRP含量降低,但心肌组织和脊髓中CGRP水平无明显变化;电针后,大鼠血浆中CGRP含量升高(P<0.01),心肌组织内CGRP含量仍无明显变化,但脊髓中CGRP含量有所升高(P<0.05)。对血浆、心脏组织及脊髓组织内的CGRP进行相关分析,未发现三者的明显相关性。结论:电针刺激具有一定的抗心肌缺血损伤作用,CGRP可能是参与电针抗心肌缺血的重要因子。     

Cardioprotective effects of saponins from Panax japonicus on acute myocardial ischemia against oxidative stress-triggered damage and cardiac cell death in rats

Aim

To study the cardioprotective effects of saponins from Panax japonicus (SPJ) on acute myocardial ischemia injury rats induced by ligating of the left anterior descending branch (LAD), on the basis of this investigation, the possible mechanism of SPJ was elucidated.

Materials and methods

SPJ was identified by high performance liquid chromatography-evaporative light scattering detection. Male Sprague-Dawley rats (200–220 g) were randomly divided into four groups: sham-operated, LAD, LAD + l-SPJ (SPJ, 50 mg/kg/day, orally) and LAD + h-SPJ (SPJ, 100 mg/kg/day, orally). Before operation, the foregoing groups were pretreated with homologous drug once a day for 7 days, respectively. After twelve hours in LAD, the cardioprotective effects of SPJ were evaluated by infarct size, biochemical values, hemodynamic, and histopathological observations and the antioxidative and antiapoptotic relative gene expressions.

Results

SPJ significantly improved heart function and decreased infarct size; remarkably decreased levels of serum lactate dehydrogenase, creatine kinase, xanthine oxide and malondialdehyde content, increased contents of serum total antioxidant capacity, superoxide dismutase (SOD), glutathione peroxidase, catalase; quantitative real-time PCR results showed that SPJ might markedly reverse the down-regulated mRNA expressions of the SOD1, SOD2 and SOD3, ameliorate the increased Bax and caspase-3 mRNA expressions and decreased Bcl-2 mRNA expression and ratios of Bcl-2 to Bax. Histopathological observations provided supportive evidence for biochemical analyses, and with the dose of SPJ increasing, the aforesaid improvement became more and more strong.

Conclusions

The studies demonstrated that in ischemic myocardium, oxidative stress caused the overgeneration and accumulation of reactive oxygen species (ROS), which was central of cardiac ischemic injury. SPJ exerted beneficially cardioprotective effects on myocardial ischemia injury rats, mainly scavenging oxidative stress-triggered overgeneration and accumulation of ROS, alleviating myocardial ischemia injury and cardiac cell death.     

淫羊藿苷对小鼠破骨细胞MMP-9、CK mRNA表达的影响

目的观察淫羊藿苷对体外培养小鼠破骨细胞MMP-9、CK mRNA表达的影响。方法用浓度分别为25 ng/ml、30 ng/ml、10-8mol/L的巨噬细胞集落刺激因子(M-CSF)、核因子κB受体活化因子配体(RANKL)、1,25(OH)2VitD3体外诱导培养小鼠骨髓源性破骨细胞,分别加入0 mol/L、10-5mol/L的淫羊藿苷,培养48 h后,抽提总RNA,采用半定量逆转录-聚合酶链反应(RT-PCR)方法检测破骨细胞中基质金属蛋白酶-9(MMP-9)、组织蛋白酶K(CK)mRNA表达的变化。结果与空白对照组比较,10-5mol/L的淫羊藿苷可明显下调MMP9mRNA的表达(P0.05),但对CK mRNA的表达无明显影响(P0.05)。结论淫羊藿可以通过下调MMP-9基因表达,从而抑制破骨细胞的分化形成及骨吸收。