硫酸乙酰肝素协同细胞因子对脐血造血细胞体外生长的作用

目的 探讨硫酸乙酰肝素（HS）协同细胞因子对脐血造血细胞体外生长的影响。方法 细胞因子（rhIL-3,rhIL-1β，rhIL-6和SCF），骨髓基质细胞培养上清（SN） 细胞因子，SN，HS 细胞因子4组分别与脐血造血细胞培养，检测细胞增殖，细胞表面标志和造血祖细胞集落，生长。结果 培养至第14天细胞增殖达高峰，4个组细胞总数依次增加了134.5,171.3,81.5和167.2倍。第21天细胞数量SN组明显下降。CD34^ 细胞数第7天已升高，第14天达高峰，依次增加了68.4%,82.5%,69.8%和79.3%，第21天仍为56.2%,71.7%,12.3%和73.7%,CD33^ 细胞数第14天达高峰，依次增加了80.2%,68.6%,81.6%和70.3%,第21天时仍维持在第14天水平，CD38^ 细胞数第7天时比培养前增加了66.6%,73.8%,70.4%和71.9%；第14至第21天均维持该水平，扩增1周各组CFU-GM产率均有提高，2周达最高，依次增加了250%，279%，217%和273%。第3周仍有151%，240%，145%和231%。结论 SN与细胞因子联用对脐血造血细胞增殖的促进作用明显强于两者单用。HS可替代SN与细胞因子联用。     

急性肺损伤肺间质巨噬细胞TLR4的表达及意义

目的 观察严重胸部创伤致急性肺损伤肺间质巨噬细胞TLR4基因及蛋白表达水平的变化,并探讨其意义。方法 建立严重胸部创伤致急性肺损伤模型。肺组织机械剪碎,然后用胶原酶、DNA酶消化肺组织,分离、培养肺间质巨噬细胞,利用免疫印迹蛋白、Northern Blot等分子生物学技术检测创伤前以及创伤后2、4、8、16和24 h肺泡巨噬细胞TLR4蛋白及基因表达。结果 利用小型多功能生物撞击机以400 kPa驱动压力对大鼠右侧上胸壁进行致伤,能够建立稳定可靠并且符合临床特点的严重胸部创伤模型;严重胸部创伤可以上调TLR4蛋白及基因在肺间质巨噬细胞内的表达,表达高峰在伤后8和16 h,伤后24 h恢复正常。结论 TLR4的上调参与了创伤后失控性炎症反应的病理生理过程。     

腹白线疝2例

病例 例1,女,50岁.腹壁肿物逐渐增大20年入院.20年前患者无意中发现腹壁肿物约核桃大小,压之可变小或消失,未予重视.20年后肿物逐渐增大至鸭蛋大小.既往育有3子,自然顺产.     

Antiatherosclerotic effects of a novel synthetic tissue-selective steroidal liver X receptor agonist in low-density lipoprotein receptor-deficient mice

Liver X receptor (LXR) agonists have the potential to treat atherosclerosis based on their ability to enhance reverse cholesterol transport. However, their side effects, such as induction of liver lipogenesis and triglyceridemia, may limit their pharmaceutical development. In contrast to the nonsteroidal LXR agonist N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide (T0901317), 3alpha, 6alpha, 24-trihydroxy-24, 24-di(trifluoromethyl)-5beta-cholane (ATI-829), a novel potent synthetic steroidal LXR agonist, was a poor inducer of sterol regulatory element-binding protein 1c expression in hepatoma HepG2 cells, whereas both compounds increased ABCA1 expression in macrophage THP-1 cells. In male low-density lipoprotein receptor-deficient mice, ATI-829 selectively activated LXR target gene expression in mouse intestines and macrophages but not in the liver. A significant increase in liver triglyceride and plasma triglyceriderich small very low-density lipoprotein (VLDL) was observed in T0901317 but not ATI-829-treated mice. Compared with vehicle-treated mice, atherosclerosis development was significantly inhibited in the innominate artery after treatment with either compound. However, in the aortic root, inhibition of atherosclerosis was only observed in the right (right coronary artery-associated sinus) but not the left coronary-related sinus (left coronary artery-associated sinus; LC) of mice treated with either compound. Lesions in the innominate artery were less complex after treatment with either compound and contained mostly macrophage foam cells. In contrast, LC lesions were more complex and had a large collagen-positive fibrous cap and less macrophage foam cell area after treatment with either compound. The T0901317-induced hypertriglyceridemia was accompanied by an increase in small triglyceride-rich VLDL that may influence LXR agonist-mediated antiatherosclerotic effects at certain vascular sites. ATI-829, by selectively activating LXR in certain tissues without inducing hypertriglyceridemia, is a good candidate for drug development.     

心力衰竭患者心功能的改善与参麦注射液及血清炎症因子水平的关系

目的:观察心力衰竭患者经参麦注射液治疗前后心功能指标及血液肿瘤坏死因子α、白细胞介素6、白细胞介素8水平的变化。方法:选择2002-03/2004-06解放军第三○九医院心血管内科住院心功能Ⅱ～Ⅳ级心力衰竭患者58例,男32例,女26例。随机将患者分为2组,西医组28例,参麦组30例。对照组给予西医一般常规治疗;参麦组在西医常规治疗的同时,加用参麦注射液静脉滴注,50g/L葡萄糖注射液500mL中加参麦注射液30mL,1次/d,治疗3周。两组治疗前后均采血测定血清中肿瘤坏死因子α、白细胞介素6、白细胞介素8水平。测定心功能指数,采用多普勒超声测定左室收缩末期容积、舒张末期容积和左室射血分数。结果:按意向处理分析,进入结果分析心力衰竭患者58例,西医组28例,参麦组30例。①两组治疗后心功能指标均较治疗前有显著改善;参麦组治疗后心功能指数、左室射血分数明显高于西医组治疗后犤(2.83±0.65)分/例,(52.7±9.1)%;(2.43±0.78)分/例,(46.8±6.6)%,t=2.258,2.579,P<0.05犦;左室收缩末期容积明显小于西医组治疗后犤(67.4±21.5),(80.6±20.4)mL,t=2.336,P<0.05犦。②两组患者治疗后血清细胞因子水平均明显降低。参麦组治疗后血清肿瘤坏死因子α、白细胞介素6、白细胞介素8水平明显低于西医组治疗后犤(74.1±12.4),(47.8±6.7),(64.3±11.8)ng/L;(85.4±11.5),(55.7±8.3),(71.3±12.8)ng/L,t=5.312,3.453,4.256,P<0.01犦。结论:应用参麦注射液干预后,心力衰竭患者血清白细胞介素6及白细胞介素8水平显著下调,说明在常规治疗的基础上使用参麦注射液可以使炎症反应减轻,参麦注射还能有效改善心力衰竭患者的心脏功能。     

互动式教学在手术室临床带教中的应用及效果分析

目的探索运用互动式教学方法提高手术室临床教学质量的效果。方法对照组35名实习大专护生采用传统教学方法,实验组32名实习大专护生采用互动式教学方法,并对两组护生的实习效果进行比较。结果两组护生对指导老师的满意度差异具有统计学意义(P<0.05);护生工作态度、工作积极性和学习积极性差异均具有统计学意义(P<0.05);老师工作积极性增加(P<0.01);两组护生的收获和对工作程序的熟悉度,以及常见手术配合的熟练度差异具有统计学意义(P<0.05);护生对手术室带教方式的满意度差异具有统计学意义(P<0.05);在提高护生无菌技术观念方面两组差异无统计学意义(P>0.05)。结论互动式教学重视护生和老师平衡发展,贯穿“以人为本”的教学理念,使人性化的教学得以体现,提高了教学质量。     

去痰活络健脑制剂改善脑缺血模型大鼠记忆功能的途径

目的:通过放射免疫法测定脑缺血模型大鼠脑组织中神经肽Y及白细胞介素2、白细胞介素6的含量,分析以去痰活络为治则的健脑制剂健脑宁改善其记忆障碍的可能途径。方法:实验于2000-07/2003-02在北京中医药大学东直门医院临床药理实验室完成。选取清洁级4月龄SD大鼠152只,取150只随机分为多发性脑梗死、中动脉脑梗死、去脑膜毛细血管造模大鼠各50只,每组模型随机再分为5组,即对照组、模型组、健脑宁11.25,3.75,1.25g/(kg·d)剂量组,每组10只,雌雄各半;余2只用于取血,配制血栓水溶液。健脑宁胶囊(由天麻、人参叶、枳壳等组成,武汉健民药业集团股份有限公司,批号:000205),实验时将药物用5g/L消毒羧甲基纤维素钠配制成混悬液。150只大鼠在无菌条件下采用由颈总动脉向颈内动脉注入同种动物血栓、结扎颈外动脉由颈外动脉向颈内动脉插线再灌注、开颅去皮质毛细血管的方法制造了3种实验性记忆功能减退实验动物模型,对照组均为假手术。术后对照组、模型组喂饲5g/L消毒羧甲基纤维素钠,健脑宁11.25,3.75,1.25g/(kg·d)剂量组,给生药剂量分别为11.25,3.75,1.25g/(kg·d),用5g/L消毒羧甲基纤维素钠配制,给药体积均为10mL/kg体质量。连续给药4周,麻醉取血,分离血浆,用放射免疫法测定中动脉脑梗死、多发性脑梗死、去皮质毛细血管模型大鼠脑组织中神经肽Y及白细胞介素2、白细胞介素6的含量。结果:每个模型组均取模型组9只,健脑宁11.25g/(kg·d)剂量组8只,3.75g/(kg·d)剂量组9只,1.25g/(kg·d)剂量组8只,对照组9只;剔除手术过程中死亡或造模不成功大鼠21只,最终129只进入结果分析。①实验4周后,多发性脑梗死、去脑膜毛细血管、中动脉脑梗死正常组大鼠海马组织中白细胞介素2含量明显高于模型组,白细胞介素6含量和血浆、海马、脑皮质、下丘脑组织中神经肽Y含量均明显低于模型组(P<0.05～0.01)。②多发性脑梗死、去脑膜毛细血管、中动脉脑梗死健脑宁11.25g/(kg·d)剂量组大鼠海马组织中白细胞介素2含量与模型组比较明显升高,白细胞介素6含量和血浆、海马、脑皮质、下丘脑组织中神经肽Y含量均明显降低(P<0.05～0.01)。③多发性脑梗死、去脑膜毛细血管健脑宁3.75g/(kg·d)剂量组大鼠海马组织中白细胞介素6、海马、脑皮质、下丘脑组织中神经肽Y含量与模型组比较明显降低,海马组织中白细胞介素2含量明显升高(P<0.05～0.01)。④去脑膜毛细血管健脑宁1.25g/(kg·d)剂量组大鼠海马组织中白细胞介素6、中动脉脑梗死大鼠血浆、下丘脑组织中神经肽Y含量与模型组比较均明显降低(P<0.05～0.01)。结论:健脑宁能调节3种脑缺血实验模型大鼠脑组织中白细胞介素2、白细胞介素6和神经肽Y的含量可能是其具有改善脑缺血模型大鼠记忆功能的重要机制。     

Pancreatic Islet Vasculature Adapts to Insulin Resistance Through Dilation and Not Angiogenesis

Pancreatic islets adapt to insulin resistance through a complex set of changes, including β-cell hyperplasia and hypertrophy. To determine if islet vascularization changes in response to insulin resistance, we investigated three independent models of insulin resistance: ob/ob, GLUT4^+/−, and mice with high-fat diet–induced obesity. Intravital blood vessel labeling and immunocytochemistry revealed a vascular plasticity in which islet vessel area was significantly increased, but intraislet vessel density was decreased as the result of insulin resistance. These vascular changes were independent of islet size and were only observed within the β-cell core but not in the islet periphery. Intraislet endothelial cell fenestration, proliferation, and islet angiogenic factor/receptor expression were unchanged in insulin-resistant compared with control mice, indicating that islet capillary expansion is mediated by dilation of preexisting vessels and not by angiogenesis. We propose that the islet capillary dilation is modulated by endothelial nitric oxide synthase via complementary signals derived from β-cells, parasympathetic nerves, and increased islet blood flow. These compensatory changes in islet vascularization may influence whether β-cells can adequately respond to insulin resistance and prevent the development of diabetes.Pancreatic islets are highly vascularized, and this feature is critical for β-cells to rapidly sense the blood glucose and secrete insulin into the systemic circulation (1,2). Islet vascularization begins early in pancreas development and is maintained in adulthood as a consequence of islet cell production of angiogenic factors such as vascular endothelial growth factor-A (VEGF-A) and angiopoietin-1 (Ang-1) (3–6). These factors recruit endothelial cells (ECs), stimulate blood vessel growth and maturation, and in the case of VEGF-A, promote formation of EC fenestrations (5,6). In addition, ECs adjacent to pancreatic epithelium reciprocally influence islet cell differentiation and development (7,8).β-Cells have a remarkable ability to respond to changes in an organism’s metabolic state, such as changes in the blood glucose or increased insulin requirements. For example, when insulin resistance develops, β-cells of the pancreatic islet can dramatically increase insulin production and secretion with an increase of β-cell mass, thus maintaining normoglycemia (9,10). In this way, mouse models with marked insulin resistance and humans with obesity-related insulin resistance are hyperinsulinemic but not hyperglycemic. The mechanisms underlying this β-cell adaptation to insulin resistance and their subsequent failure in some individuals who develop type 2 diabetes are incompletely understood.Because of the highly vascularized state of pancreatic islets and the marked changes in β-cell size and number in the setting of insulin resistance, we hypothesized that the islet vasculature must adapt to these changes in β-cell mass and insulin requirements. We envisioned that a hyperplastic islet, like a growing tumor mass, would increase production of angiogenic factors to increase its vascular supply with expanding β-cell mass (11). To test this hypothesis, we examined islet vascularization in three mouse models of insulin resistance and found, unexpectedly, that islet vessel density was decreased, not increased, and that the intraislet vasculature became markedly dilated whereas vessels in the exocrine tissue were unchanged. The dilation of intraislet capillaries was independent of islet size, suggesting the vascular adaptation may primarily support increased β-cell insulin secretory demand rather than β-cell mass expansion. Moreover, these vascular changes were accompanied by an increase in islet parasympathetic innervation. Our results indicate that the metabolic state influences islet angioarchitecture and innervation, suggesting that islet neurovascular remodeling may influence whether β-cells can adequately respond to insulin resistance and maintain normoglycemia.     

不同剂量维生素A强化饼干对3～6岁儿童维生素A营养状况的干预效果

背景:维生素A缺乏性疾病是一个世界性的儿童健康问题,中国西部及边远地区尤为严重,需要进行营养干预。目的:比较补充3种不同剂量强化维生素A饼干对3～6岁儿童维生素A营养状况的改善效果,探索预防维生素A缺乏性疾病的强化饼干的理想剂量。设计:随机对照观察。单位:重庆市卫生局卫生监督所、重庆医科大学公共卫生学院营养与食品卫生学教研室和重庆医科大学儿童医院儿童营养研究中心。对象:调查于2002-03/12完成,选择重庆市巴南区8个幼儿园的3～6岁儿童753名,监护人均知情同意。随机分为4组,即30%推荐摄入量组、100%推荐摄入量组、2万国际单位组及20万国际单位组。方法:①30%推荐摄入量组(177名)每日补充维生素A30%每日推荐摄入量(500IU)强化饼干1次。②100%推荐摄入量组(173名)每日补充维生素A100%每日推荐摄入量(1666IU)强化饼干1次。③2万国际单位组(209名)每周补充维生素A2万国际单位强化饼干1次。④20万国际单位组(194名)补充维生素A20万国际单位胶丸1次。所有儿童干预前及干预3个月时测定身高、体质量、血清视黄醇、前白蛋白、血红蛋白及视黄醇结合蛋白。30%推荐摄入量组中有87名儿童在补充9个月后再次复查以上指标。主要观察指标:①各组儿童干预前及干预3个月后维生素A缺乏性疾病检出率。②各组儿童干预前及干预3个月后血清视黄醇、血清前白蛋白、血清视黄醇结合蛋白、血红蛋白、身高和体质量。结果:因样本流失和检测技术的问题,复查仅得580名儿童的检测结果。①各组儿童维生素A缺乏性疾病检出率比较:补充3个月后,30%推荐摄入量组、100%推荐摄入量组、2万国际单位组及20万国际单位组儿童维生素A缺乏性疾病检出率均较治疗前显著下降[1.48%,1.42%,1.21%,2.16%;6.78%,6.54%,8.61%,8.25%(χ2=3.86～8.57,P<0.05～0.01)]。②各组儿童血清视黄醇、血清前白蛋白、血清维生素A、血红蛋白、身高和体质量比较:补充3个月后,自身对照除30%推荐摄入量组的前白蛋白和血红蛋白外,各组儿童其余各项指标均明显增加(t=2.52～37.44,P<0.05～0.01)。2万国际单位组血清维生素A的升高幅度明显大于其他组(F=4.62,P<0.01),30%推荐摄入量组血红蛋白、前白蛋白和身高的增长幅度明显小于其他组(F=5.0～7.78,P<0.01)。30%推荐摄入量组补充9个月后,血红蛋白和前白蛋白的升高明显大于其余组(F=11.62,10.21,P<0.01),血清视黄醇的升高仍低于2万国际单位组(F=4.21,P<0.01)。结论:补充3种不同剂量的维生素A强化饼干和20万国际单位维生素A胶丸,均可明显改善维生素A的营养状况和血红蛋白水平。其中30%推荐摄入量和100%推荐摄入量风险小、且每日补充能稳定维持体内维生素A水平,可能是更适于儿童的长期补充剂量,而30%推荐摄入最优于100%推荐摄入量。     

大鼠骨髓和脂肪来源间充质干细胞的成骨特性比较

背景:尽管骨髓来源和脂肪来源的间充质干细胞均具有向骨和软骨分化潜能,但终究带有各自来源组织的特点,那么是否意味着两者在非基因影响下成骨特性存在一定差异呢?目的:比较大鼠骨髓和脂肪来源的间充质干细胞在体外成骨分化中的特性区别。设计、时间及地点:细胞学体外对照观察,于2007-07/2008-03在中国科学院昆明动物研究所国家级重点实验室完成。材料:6周龄和18周龄SD大鼠各2只,分别用于制备骨髓间充质干细胞和脂肪间充质干细胞。方法:选取第3代骨髓间充质干细胞和脂肪间充质干细胞,按5×107L-1接种后各分为2组:诱导组加入含10-8mol/L地塞米松、10mol/Lβ-甘油磷酸、50mg/L维生素C的成骨诱导培养基1.5mL,未诱导组不加入成骨诱导培养基。主要观察指标:碱性磷酸酶染色与茜素红染色结果,碱性磷酸酶和骨钙素定量分析结果。结果:成骨诱导7,14d,经诱导的骨髓基质干细胞和脂肪间充质干细胞呈碱性磷酸酶阳性,有钙结节形成,未诱导组呈阴性。成骨诱导3,7,10d,未诱导组间比较碱性磷酸酶及骨钙素含量均基本相似(P>0.05);与未诱导组比较,脂肪间充质干细胞诱导组碱性磷酸酶及骨钙素含量均明显升高(P<0.05),骨髓间充质干细胞诱导组升高幅度更为显著(P<0.001)。结论:大鼠骨髓来源间充质干细胞体外成骨性能强于脂肪来源间充质干细胞,在骨组织工程种子细胞的选用中应优先考虑前者。