人脂肪干细胞体外培养特性及成脂成软骨分化

目的:观察人脂肪组织间充质干细胞体外分离培养及成脂、成软骨诱导分化的生物学性状,探讨其作为软骨组织工程种子细胞的可行性。方法:实验于2006-08/2007-03在吉林大学中日联谊医院前列腺疾病防治研究中心完成主要工作。实验方法:①人脂肪组织来源于健康成年女性腹部吸脂术,酶消化法分离出脂肪间充质干细胞,接种于含体积分数为0.10胎牛血清的高糖DMEM培养基进行原代培养。②取第3代脂肪间充质干细胞,采用细胞计数法测定细胞生长曲线,流式细胞仪测定CD13、CD34抗原的表达,免疫荧光法测定CD34抗原的表达。③取第3代脂肪间充质干细胞,用含体积分数为0.10胎牛血清、0.5mmol/LIBMX、1μmol/L地塞米松、10μmol/L胰岛素的成脂诱导培养基和含体积分数为0.01胎牛血清、10μg/L转化生长因子β1、50nmol/L抗坏血酸、6.25mg/L胰岛素的成软骨诱导培养基分别诱导分化。④每天用倒置显微镜观察细胞形态及增殖变化,油红"O"、AB-PSA染色和Ⅱ型胶原免疫细胞化学染色检测成脂、成软骨分化情况。结果:①体外培养的脂肪间充质干细胞呈扁平的长梭形,细胞形态均一,传代稳定。间质细胞相关标志CD13和干细胞相关标志CD34表达阳性。②定向诱导后表现出脂肪细胞和软骨细胞特性。经成脂诱导,细胞内出现空泡,油红"O"染色呈红色;经成软骨染色,AB-PSA染色呈紫红色,Ⅱ型胶原免疫细胞化学染色胞浆呈棕黄色。结论:脂肪干细胞能向软骨细胞方向诱导分化,可作为软骨组织工程种子细胞。     

低氧运动对SD大鼠白细胞及其分类计数的影响

目的:观察低氧及运动对大鼠白细胞及其分类计数的影响。方法:实验于2005-03/05在广州体育学院运动生化省重点实验室进行。随机选择34只喜好运动,能在跑台上自觉跑步的SD大鼠数字表法随机分为常氧安静组、常氧运动组、低氧安静组、低氧运动组4组,常氧运动组在常氧状态下(氧的体积分数是0.209)进行25m/min的跑台训练,低氧运动组大鼠常压低氧环境(氧的体积分数是0.154,相当于2500m高度)下进行20m/min的跑台训练,训练时间均为1h/d,6d/周,持续4周。常氧安静组和低氧安静组分别在常氧和低氧环境中4周,不运动。4周后大鼠禁食12h,腹主动脉取血,采用德国Bayer公司的ADVIA120全自动血球分析仪测定血液中白细胞总数、中性粒细胞、淋巴细胞、单核细胞、嗜酸细胞、嗜碱细胞计数。结果:34只大鼠全部进入结果分析。①白细胞总数和淋巴细胞各组间比较差异不显著(P>0.05)。②中性粒细胞计数:低氧安静组和低氧运动组低于常氧运动组[(0.15±0.05)×103,(0.14±0.10)×103,(0.35±0.39)×103L-1,P<0.05,0.01]。③单核细胞计数:低氧安静组和低氧运动组低于常氧安静组[(0.48±0.32)×103,(0.53±0.30)×103,(1.08±0.53)×103L-1,P<0.05,0.01]。④嗜酸细胞计数:常氧运动组、低氧安静组和低氧运动组低于常氧安静组[(0.008±0.009)×103,(0.007±0.008)×103,(0.013±0.009)×103,(0.047±0.029)×103L-1,P<0.01]。⑤嗜碱细胞计数:低氧安静组和低氧运动组低于常氧运动组[(0.007±0.005)×103,(0.006±0.005)×103,(0.018±0.015)×103L-1,P<0.05]。结论:长期的低氧适应能显著性降低运动大鼠中性粒细胞反应性,并显著性降低安静大鼠的单核细胞及嗜酸细胞数,造成机体部分非特异性免疫力有所下降。     

A randomized trial on the efficacy of an autologous blood drainage and transfusion device in patients undergoing elective knee arthroplasty

The purpose of the study reported here was the determination of the efficacy of a postoperative autologous blood drainage and transfusion device in reducing allogeneic red cell requirements in patients undergoing elective knee arthroplasty. The study was a randomized controlled trial with adult patients undergoing unilateral elective arthroplastic knee surgery. Patients underwent suction drainage, attached to an autologous blood drainage and transfusion device, or standard suction drainage. Allogeneic red cells were given according to strict transfusion guidelines based on blood loss and postoperative hemoglobin values. Outcome measures included the mean number of allogeneic red cell concentrates required and the number of patients in each group who required no transfusion. Patients assigned to standard suction drainage had a mean allogeneic red cell utilization of 1.2 units (SD 1.0), as compared to a mean of 0.4 units (SD 0.8) in the group undergoing drainage with the autologous blood drainage and transfusion device (p = 0.0007). The percentage of patients not requiring allogeneic red cells was significantly higher in the latter group (74.3% vs. 32.5%; p = 0.002). The postoperative drainage and transfusion device was efficacious in reducing the amount of allogeneic red cells required by patients undergoing knee arthroplasty, and its use resulted in a 42 percent reduction in the number of patients requiring allogeneic transfusion.     

HIF-1α-independent hypoxia-induced rapid PTK6 stabilization is associated with increased motility and invasion

PTK6/Brk is a non-receptor tyrosine kinase overexpressed in cancer. Here we demonstrate that cytosolic PTK6 is rapidly and robustly induced in response to hypoxic conditions in a HIF-1-independent manner. Furthermore, a proportion of hypoxic PTK6 subsequently re-localized to the cell membrane. We observed that the rapid stabilization of PTK6 is associated with a decrease in PTK6 ubiquitylation and we have identified c-Cbl as a putative PTK6 E3 ligase in normoxia. The consequences of hypoxia-induced PTK6 stabilization and subcellular re-localization to the plasma membrane include increased cell motility and invasion, suggesting PTK6 targeting as a therapeutic approach to reduce hypoxia-regulated metastatic potential. This could have particular significance for breast cancer patients with triple negative disease.     

结核性胸液淋巴细胞行为的单细胞定量研究

目的:为探讨结核性胸液淋巴细胞行为的特点及在病理过程中的意义.方法:采用单细胞微管吸吮技术,分别检测正常人外周血淋巴细胞、结核性胸膜炎患者外周血淋巴细胞、胸液中淋巴细胞与肺血管内皮细胞的粘附特点.结果:结核性胸膜炎患者外周血淋巴细胞的粘附性(2.83±0.43Pa)较正常人(1.21±0.25Pa)明显增强,但结核性胸液中淋巴细胞的粘附性(1.67±0.39Pa)却与正常人无明显差别.结论:结核性胸膜炎患者外周血淋巴细胞处刺激激活状态,其粘附性升高有利于贴壁向血管外游走进而向胸膜腔渗出.结核性胸液中淋巴细胞粘附性呈现正常水平可能系由于胸液中高浓度游离型上皮来源的粘附分子将其表面表达的粘附分子饱和的缘故.此现象提示胸液中的淋巴细胞,不易回吸收,其去除多以凋亡方式,最好籍助外力清除(胸穿抽液).     

硒对高硫所致豚鼠心肌退变的保护作用

豚鼠体内含硫量的增加，可使心肌发生不同程度的退变，如果体内含硒量也增加，心肌则不发生退变。高硫能抑制机体摄取硒的能力，使体内含硒量下降。心肌退变的程度与体内含硒量呈负相关。提示：高硫对心肌的损伤是通过低硒来完成的，高硫只是诱发低硒形成的条件；心肌退变可能与血硒和血硫是否维持在一定的比值范围内有关。     

p16基因对前列腺癌细胞的体外作用研究

目的：研究可控性表达的p16基因对前列腺癌细胞的生长抑制作用。方法：通过可诱导的真核表达载体pMDNA-3-p16将外源野生型p16基因转染至该基因表达功能丧失的人前列腺癌细胞PC3中,使用ZnSO4作为诱导物,观察p16基因的可控性表达及其对前列腺癌细胞生物学特性的影响。 结果：转染了野生型p16基因的人膀胱癌细胞PC3-pMDNA3-p16经ZnSO4诱导后,开放了p16基因的转录和翻译,p16蛋白得到高水平表达,细胞生长速度及集落形成率均有了部分抑制,细胞周期分析可见G1期增加,S期下降。结论：P16基因与前列腺癌的发生、发展有关,为用p16基因进行前列腺癌的基因治疗提供了实验依据。