B1 but not B2 bradykinin receptor agonists promote DU145 prostate cancer cell proliferation and migration

Background

The kallikrein-kinin system (KKS) is an endogenous pathway involved in angiogenesis and tumourigenesis, both vital for cancer growth and progression.

Objectives

To investigate the effect of two bradykinin receptor (B1R and B2R) agonists on growth and motility of prostate tumour (DU145) and micro-vascular endothelial cells (dMVECs).

Methods

Increasing concentrations of selective B1R and B2R agonists were added to cultured cells. Cell proliferation and migration were assessed using the 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) and modified Boyden Chamber assays, respectively. Where significant stimulation was found, the influence of an antagonist was also investigated.

Results

Neither growth nor motility of endothelial cells was affected by either agonist. In DU145 cells, while the B2R agonist was without any significant effect, the B1R agonist stimulated proliferation and migration at concentrations of 10nM and 50nM respectively. Further, this effect was abrogated when cells were pre-incubated with a B1R antagonist.

Conclusions

Unlike the physiologically-active B2R, the pathologically-inducible B1R may be implicated in prostate tumourigenic events. The involvement of the KKS in malignant prostate pathology supports on-going exploration of bradykinin receptor antagonists as target candidates in the development of alternate approaches to cancer therapy.     

Identification of Candida species in the clinical laboratory: a review of conventional,commercial, and molecular techniques

In healthy individuals, Candida species are considered commensal yeasts of the oral cavity. However, these microorganisms can also act as opportunist pathogens, particularly the so‐called non‐albicans Candida species that are increasingly recognized as important agents of human infection. Several surveys have documented increased rates of C. glabrata, C. tropicalis, C. guilliermondii, C. dubliniensis, C. parapsilosis, and C. krusei in local and systemic fungal infections. Some of these species are resistant to antifungal agents. Consequently, rapid and correct identification of species can play an important role in the management of candidiasis. Conventional methods for identification of Candida species are based on morphological and physiological attributes. However, accurate identification of all isolates from clinical samples is often complex and time‐consuming. Hence, several manual and automated rapid commercial systems for identifying these organisms have been developed, some of which may have significant sensitivity issues. To overcome these limitations, newer molecular typing techniques have been developed that allow accurate and rapid identification of Candida species. This study reviewed the current state of identification methods for yeasts, particularly Candida species.     

L-精氨酸对糖尿病大鼠心肌缺血再灌注损伤的影响

目的:观察糖尿病大鼠心肌缺血再灌注时血管紧张素Ⅱ、胰岛素样生长因子1、醛固酮、细胞间黏附分子1和自由基代谢的变化及L-精氨酸对其的影响。方法:实验于2005-02/2006-06在江苏大学医学院机能学实验室完成。①实验分组:腹腔注射链脲佐菌素制作糖尿病大鼠模型,30只大鼠造模成功。按随机数字表法分为3组(n=10):心肌缺血再灌注组:开胸结扎冠脉,造成心肌缺血,60min后放松再灌注60min;L-精氨酸治疗组:于手术前4周灌胃L-精氨酸250mg/(kg·d),然后重复心肌缺血再灌注组操作;假手术组:完成操作后只穿线不结扎,观察2h作为对照。实验结束时心室取血6mL,摘取心脏,留取左心室心肌组织。②实验评估:检测大鼠血浆血管紧张素Ⅱ、醛固酮和血清胰岛素样生长因子1含量及心肌细胞间黏附分子1蛋白表达。检测大鼠血清、心肌组织超氧化物歧化酶、谷胱甘肽-过氧化物酶活性、丙二醛含量及心肌线粒体Na ,K -ATP酶、Mg2 -ATP酶、Ca2 -ATP酶活性。结果:30只大鼠全部进入结果分析。①与假手术组相比,心肌缺血再灌注组血浆血管紧张素Ⅱ、醛固酮含量明显升高(P<0.05～0.01),血清胰岛素样生长因子1含量降低(P<0.05);L-精氨酸治疗4周后血浆血管紧张素Ⅱ、醛固酮含量低于心肌缺血再灌注组(P<0.05～0.01),血清胰岛素样生长因子1含量高于心肌缺血再灌注组(P<0.05)。②与假手术组相比,心肌缺血再灌注组血清、心肌丙二醛含量明显升高(P<0.05),血清、心肌超氧化物歧化酶和谷胱甘肽-过氧化物酶活性明显降低(P<0.05￣0.01);用L-精氨酸治疗4周后血清、心肌丙二醛含量低于心肌缺血再灌注组(P<0.05￣0.01),血清、心肌超氧化物歧化酶和谷胱甘肽-过氧化物酶活性高于心肌缺血再灌注组(P<0.05～0.01)。③与假手术组相比,心肌缺血再灌注组心肌线粒体Na ,K -ATP酶、Mg2 -ATP酶、Ca2 -ATP酶活性明显降低(P<0.05),心肌细胞间黏附分子1蛋白表达明显升高(P<0.01);用L-精氨酸治疗4周后心肌线粒体Na ,K -ATP酶、Mg2 -ATP酶、Ca2 -ATP酶活性明显高于心肌缺血再灌注组(P<0.05),心肌细胞间黏附分子1蛋白表达低于心肌缺血再灌注组(P<0.05)。结论:血管紧张素Ⅱ、醛固酮和胰岛素样生长因子1可能共同参与了糖尿病心肌缺血再灌注的发生,细胞间黏附分子1蛋白表达与糖尿病心肌损伤关系密切。L-精氨酸通过减少细胞间黏附分子1蛋白表达,起心肌保护作用。糖尿病心肌缺血再灌注时存在自由基代谢异常,补充L-精氨酸后,可通过提高超氧化物歧化酶、谷胱甘肽-过氧化物酶和ATP酶活性,降低丙二醛水平,减轻自由基损伤,改善心肌组织功能。     

四种中成药对气血双虚模型小鼠血象及免疫水平的影响

目的:为艾滋病抗病毒疗法所致的骨髓不良反应筛选疗效确切的中成药,观察分析参芪颗粒、复方阿胶浆、贞芪扶正颗粒、复方皂矾丸四种中成药对放血和注射环磷酰胺联合复制的气血双虚模型小鼠血象及免疫水平的影响。方法:实验于2005-08/09在河南中医学院药理实验室完成。①参芪颗粒(江西山高制药有限公司生产,批号040702);复方阿胶浆(山东东阿阿胶股份有限公司生产,批号050446);贞芪扶正颗粒(甘肃扶正药业科技股份有限公司生产,批号040803);复方皂矾丸(陕西郝其军制药有限责任公司生产,批号041014);当归补血口服液(郑州市协和制药厂生产,批号041122);环磷酰胺(上海华联制药有限公司生产,批号050101)。②选取清洁级昆明种小鼠150只,随机数字表法分为15组,10只/组:1～3组分别灌服参芪颗粒混悬液3,2,1g/kg;4～6组分别灌服复方阿胶浆30,20,10mL/kg;7～9组分别灌服贞芪扶正颗粒混悬液15,10,5g/kg;10～12组分别灌服复方皂矾丸混悬液2.4,1.6,0.8g/kg;第13组灌服当归补血口服液10g/kg;剩余2组为空白对照组和模型对照组,分别给于同体积生理盐水10g/kg。各组给药1次/d,连续给药10d。③除空白对照组外,其他各组从给药第1天开始建立气血双虚模型。每只鼠尾部放血0.25mL/10g,然后分别于第2,4,6,8天腹腔注射环磷酰胺80,40,40,40mg/kg。空白对照组同时间点仅腹腔注射等体积生理盐水。末次注射环磷酰胺后2h,眼眶取血,一部分用于血象测定,另一部分离心取血清,测定血细胞比容及血清中巨噬细胞集落刺激因子水平;解剖取胸腺和脾脏,检测胸腺皮质厚度、胸腺淋巴细胞数、脾小结大小、脾脏淋巴细胞数病理学指标的变化。结果:150只小鼠全部进入结果分析,放血和注射环磷酰胺并用可成功建立小鼠气血双虚模型。①与模型对照组比较,参芪颗粒3g/kg组、贞芪扶正颗粒10,5g/kg组、复方皂矾丸1.6g/kg组均可升高气血双虚模型小鼠白细胞水平(t=2.18～2.74,P<0.05),贞芪扶正颗粒15g/kg组作用更为显著(t=2.98,P<0.01);参芪颗粒1g/kg组、复方阿胶浆20mL/kg组、贞芪扶正颗粒15,10g/kg组均可升高红细胞水平(t=2.44～2.69,P<0.05),复方阿胶浆30mL/kg组、贞芪扶正颗粒5g/kg组、复方皂矾丸2.4,1.6g/kg组作用更为显著(t=2.91～3.66,P<0.01);当归补血口服液组、复方阿胶浆20mL/kg组、参芪颗粒3,1g/kg组、贞芪扶正颗粒15,10,5g/kg组均可升高血红蛋白水平(t=2.27～2.85,P<0.05),复方阿胶浆30mL/kg组、复方皂矾丸2.4,1.6g/kg组作用更为显著(t=3.07～4.04,P<0.01);当归补血口服液组、参芪颗粒3,2g/kg组均可升高血小板水平(t=2.20～2.41,P<0.05)。②与模型对照组比较,参芪颗粒2g/kg组、贞芪扶正颗粒5g/kg组均可升高气血双虚模型小鼠血细胞比容(t=2.01～2.62,P<0.05),参芪颗粒1g/kg组、复方阿胶浆30,20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4,1.6,0.8g/kg组作用更为显著(t=3.18～4.36,P<0.01);参芪颗粒2g/kg组、复方阿胶浆30,20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4,1.6g/kg组均可显著升高巨噬细胞集落刺激因子水平(t=3.60～6.80,P<0.01)。③与模型对照组比较,当归补血口服液组、参芪颗粒3,2,1g/kg组、复方阿胶浆30,20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4,1.6,0.8g/kg组均可显著增加气血双虚模型小鼠胸腺皮质厚度(t=3.71～9.34,P<0.01),增大脾小结(t=3.36～11.97,P<0.01),增加脾脏淋巴细胞数(t=4.29～10.44,P<0.01);复方阿胶浆30mL/kg组可明显增加小鼠胸腺淋巴细胞数(t=2.45,P<0.05),当归补血口服液组、参芪颗粒3,2,1g/kg组、复方阿胶浆20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4g/kg组作用更为显著(t=3.22～8.20,P<0.01)。结论:①四种中成药对气血双虚模型小鼠血红蛋白升高作用相近,以复方阿胶浆和贞芪扶正颗粒对白细胞和红细胞水平升高作用为强,以复方阿胶浆和贞芪扶正颗粒对血小板水平升高作用为优。②四种中成药对气血双虚模型小鼠血细胞比容的影响无差异,以贞芪扶正颗粒和复方皂矾丸对巨噬细胞集落刺激因子水平的升高作用为优。③以参芪颗粒、复方阿胶浆、贞芪扶正颗粒对胸腺皮质厚度和淋巴细胞数的促进作用为优,以参芪颗粒、贞芪扶正颗粒、复方皂矾丸对脾小结和脾脏淋巴细胞数的促进作用为优。     

免疫磁珠分选骨髓衍生肝干细胞亚群c-Kit+lin-

目的:利用免疫磁性细胞分选系统分离纯化骨髓衍生肝干细胞亚群c-Kit lin-。方法:实验于2006-07/08在南方医科大学实验动物中心完成。6～8周龄的SPF级纯系BALB/C雄性小鼠10只,体质量18～20g。收集小鼠股骨骨髓细胞,利用免疫磁性细胞分选系统,通过两步法分选纯化c-Kit lin-:将获取的lin-细胞悬液8℃条件下1500r/min离心10min,弃上清,按80μL/107加入Buffer重悬细胞。按20μL/107加生物素抗体磁珠,混匀,4℃冰箱孵育15min,按1mL/107加入Buffer洗细胞1次,8℃条件下1500r/min离心10min,弃上清,按500μL/108加入Buffer重悬细胞。Buffer500μL润MS柱,悬液过柱后,Buffer500μL/次洗柱3次,柱子脱离磁场,加1mLBuffer,用配套柱塞推出柱中的c-Kit lin-细胞,收集到c-kit lin-细胞,细胞计数。取2.0×106个细胞分成10等份,流式细胞仪分析c-Kit lin-细胞纯度,计算回收率,评估纯化效率,苔盼兰染色检测纯化前后的细胞活力。计算活细胞的百分率。细胞纯度和细胞回收率的计算:细胞纯度=分离产物中的阳性细胞数/分离细胞的总细胞数×100%,细胞回收率=分离产物中的阳性细胞数/起始标本阳性细胞总数×100%。结果:10只小鼠均进入结果分析。利用免疫磁性细胞分选系统分选出的骨髓衍生肝干细胞亚群c-Kit lin-细胞纯度和回收率分别为(77.98±2.34)%,75.40%,纯化前后细胞活力不受影响。结论:免疫磁性细胞分选系统能有效分选骨髓衍生肝干细胞亚群c-Kit lin-,纯度和回收率高,且不影响细胞活力。     

白藜芦醇对一次性力竭游泳大鼠肝脏组织保护作用的量效关系

目的:观察白藜芦醇对一次性力竭游泳大鼠肝脏组织的作用及发挥作用的最佳口服剂量。方法:实验于2006-05/07在成都体育学院运动医学系动物实验室完成。①实验分组:选取雄性SD大鼠70只,随机分为7组,每组10只,分别为安静对照组,运动对照组,运动 15mg/kg白藜芦醇组,运动 50mg/kg白藜芦醇组,运动 100mg/kg白藜芦醇组,运动 200mg/kg白藜芦醇组,运动 300mg/kg白藜芦醇组。②实验干预:不同剂量白藜芦醇组每天灌胃15,50,100,200,300mg/kg白藜芦醇,安静对照组和运动对照组分别灌胃相同体积的溶媒(二甲亚砜 生理盐水),连续5周。末次给予实验用样品1h后,各运动组每只鼠尾跟部负荷3%体质量铅皮,置于水深50cm、水温(31±1)℃游泳槽中游泳。游泳力竭后即刻,股动脉取血并迅速取出肝组织。③指标检测:赖氏比色法测定血清中谷丙转氨酶活性;邻苯三酚自氧化法测定肝组织超氧化物歧化酶活性;硫代巴比妥酸法测定肝组织丙二醛含量。结果:纳入动物70只,均进入结果分析。①血清谷丙转氨酶活性和肝组织中丙二醛含量:运动对照组显著高于安静对照组,不同剂量白藜芦醇组低于运动对照组(P<0.05或P<0.01)。运动 100,200,300mg/kg白藜芦醇组低于运动 15,50mg/kg白藜芦醇组[谷丙转氨酶活性:(972.36±121.86),(944.36±105.35),(888.34±88.68),(1773.52±89.35),(1377.78±27.01)nkat/L,P<0.01;丙二醛含量:(7.90±2.56),(7.69±3.69),(7.13±2.62),(19.90±2.21),(12.16±1.78)μmol/g,P<0.05]。100,200,300mg/kg白藜芦醇组间差异无显著性。②肝脏组织中超氧化物歧化酶活性:运动对照组显著低于安静对照组,不同剂量白藜芦醇组高于运动对照组(P<0.05或P<0.01)。运动 100,200,300mg/kg白藜芦醇组高于15,50mg/kg白藜芦醇组[(2325.80±163.37),(2379.14±121.86),(2447.16±89.18),(1096.05±120.19),(1514.64±28.17)μkat/g,P<0.01]。结论:①白藜芦醇对力竭性运动大鼠肝脏组织具有保护作用。②100,200,300mg/kg白藜芦醇对肝脏组织发挥保护作用效果优于15,50mg/kg,建议使用100mg/kg白藜芦醇就能达到理想效果。     

Study on liver injury models induced by CCl4 D-Gal and ANIT in mice

ＳｔｕｄｙｏｎｌｉｖｅｒｉｎｊｕｒｙｍｏｄｅｌｓｉｎｄｕｃｅｄｂｙＣＣｌ４ＤＧａｌａｎｄＡＮＩＴｉｎｍｉｃｅＹＡＮＧＸｉｎＢｏ１，ＨＵＡＮＧＺｈｅｎｇＭｉｎｇ１，ＣＡＯＷｅｎＢｉｎ１，ＺＨＥＮＧＭｉｎｇ１，ＣＨＥＮＨｏｎｇＹａｎ１，ＺＨＡＮ...     

Effects of sera from burn patients on human hepatocytic viscoelasticity

ＥｆｅｃｔｓｏｆｓｅｒａｆｒｏｍｂｕｒｎｐａｔｉｅｎｔｓｏｎｈｕｍａｎｈｅｐａｔｏｃｙｔｉｃｖｉｓｃｏｅｌａｓｔｉｃｉｔｙＷＡＮＧＸｉａｏＪｕｎ，ＬＵＯＸｉａｎｇＤｏｎｇ，ＬＵＯＱｉｎａｎｄＹＡＮＧＺｏｎｇＣｈｅｎｇＢｕｒｎＲｅｓｅａｒｃｈＩｎ...