胶体磷酸铬^32P关节腔内注射改善佐剂型关节炎大鼠的相关指标

目的:观察胶体磷酸铬32P关节腔内注射治疗大鼠佐剂型关节炎的效果。方法:实验于2006-07/09在南京市第一医院动物实验室完成。选择6～8周龄清洁级SD雌性大鼠30只,按随机数字表法分为3组,正常对照组、模型组、胶体磷酸铬32P治疗组,每组10只。大鼠左足跖皮内注射完全弗氏佐剂0.1mL免疫法制备佐剂型关节炎模型。胶体磷酸铬32P治疗组于造模后10d左踝关节腔内注射37GBq/L胶体磷酸铬32P0.02mL,即0.74MBq,正常对照组和模型组分别给予等量生理盐水左踝关节腔内注射。①每2周观察1次大鼠左踝关节左右径宽度。②关节炎指数评定采用关节炎评分法(0～4分),分数越高,症状越重。③于用药后2,4,6周采用99Tcm-MDP显像感兴趣区分析法计算大鼠左踝关节区和右胫腓骨的放射性计数比。④于用药后4,6周测定血清肿瘤坏死因子和白细胞介素1β水平。⑤于用药后4,6周观察大鼠滑膜组织和软骨组织病理学改变。结果:纳入大鼠30只,均进入结果分析。①用药后2周模型组大鼠左踝关节左右径宽度大于正常对照组[分别为(7.82±0.36),(5.89±0.35)mm],差异有显著性意义(t=12.16,P<0.001),胶体磷酸铬32P治疗组大鼠左踝关节左右径宽度大于模型组,差异无显著性意义(P>0.05)。用药后6周胶体磷酸铬32P治疗组大鼠左踝关节左右径宽度小于模型组[分别为(6.87±0.27),(7.25±0.26)mm],差异有显著性意义(t=2.87,P<0.05)。②用药后2周和4周胶体磷酸铬32P治疗组大鼠关节炎指数高于模型组,用药后6周胶体磷酸铬32P治疗组大鼠关节炎指数低于模型组,两组间差异均无显著性意义(P>0.05)。③用药后2周模型组大鼠感兴趣区放射性计数比高于正常对照组(分别为2.05±0.20,1.46±0.15),差异有显著性意义(t=7.46,P<0.001)。用药后6周胶体磷酸铬32P治疗组大鼠感兴趣区放射性计数比低于模型组(分别为1.52±0.18,1.78±0.24),差异有显著性意义(t=2.45,P<0.05)。④用药后4,6周模型组大鼠血清肿瘤坏死因子和白细胞介素1β水平高于正常对照组,差异有显著性意义[用药后4周分别为(2.039±0.344),(1.115±0.192)μg/L;(0.305±0.034),(0.192±0.041)μg/L,t=7.42,6.71,P<0.001。用药后6周分别为(1.694±0.305),(1.126±0.256)μg/L;(0.259±0.027),(0.191±0.019)μg/L,t=4.03,5.83,P<0.01,0.001]。用药后4,6周胶体磷酸铬32P治疗组在血清肿瘤坏死因子和白细胞介素1β水平与模型组比较,差异无显著性意义(P>0.05)。⑤用药后4周胶体磷酸铬32P治疗组和模型组滑膜组织增生和炎症细胞浸润均较明显;用药后6周胶体磷酸铬32P治疗组与正常对照组比较仍有滑膜组织增生和炎症细胞浸润,与模型组比较滑膜组织增生程度明显减轻,而炎症细胞浸润程度稍轻。用药后6周胶体磷酸铬32P治疗组大鼠左踝关节的软骨组织未见有异常改变。结论:胶体磷酸铬32P关节腔注射可减轻完全弗氏佐剂免疫大鼠受注射关节的滑膜增生程度,改善关节肿胀症状,疗效肯定。     

脂肪干细胞在体外特定培养液中向软骨细胞表型的分化

目的:应用转化生长因子β1,体外诱导脂肪干细胞向成软骨细胞表型分化,探讨其作为组织工程化软骨种子细胞的可行性。方法:实验于2005-04/2006-06在华中科技大学同济医学院公卫实验室完成。①取大鼠腹股沟处脂肪,酶消化法分离、培养脂肪干细胞,体外传代培养。②取第3代细胞通过转化生长因子β1、地塞米松和维生素C诱导脂肪干细胞向软骨细胞分化。③诱导后14d观察细胞形态变化,进行阿辛蓝染色检测软骨基质的分泌、免疫组织化学检测细胞Ⅱ型胶原的表达,采用Western-blot和反转录-聚合酶链反应检测诱导前后成软骨相关的Sox9,蛋白聚糖与Ⅱ型胶原的表达。结果:①细胞接种的最初几日,细胞呈圆形,1周后贴壁细胞呈长梭形,体积增大;14d后贴壁生长细胞基本长满单层,中心细胞排列紧密,形态与骨髓间充质干细胞相似。诱导培养后,细胞形态逐渐由梭型向多角形、多边形转变。诱导14d后多数细胞呈平坦的多边角形状细胞;其夹杂多角突起状或多角纺锤状细胞。②诱导后阿辛蓝染色示糖胺聚糖均匀分布于基质中。③免疫组织化学染色示基质中Ⅱ型胶原表达阳性。④反转录-聚合酶链反应检测成软骨相关的Sox9、蛋白聚糖、Ⅱ型胶原mRNA表达阳性。⑤Western-blot印迹检测细胞诱导后Ⅱ型胶原蛋白表达阳性。结论:脂肪干细胞在特定培养液的诱导下可向成软骨细胞表型分化,并能分泌软骨细胞特异性基质,有望成为软骨组织工程新的细胞来源。     

中药癌痛克对人肝癌细胞HepG2增殖、凋亡及Rb基因表达的影响

目的:通过观察不同浓度癌痛克对肝癌细胞株HepG2增殖及凋亡的作用,以及在相应状态下细胞内Rb基因表达量的改变,探讨中药癌痛克抗肝癌的可能作用机制。方法:实验于2005-09/2006-03在广州医学院中心实验室完成。取对数生长期的HepG2细胞,使细胞静止于G0/G1期。随机分为癌痛克2,10,50mg/L组和对照组,癌痛克2,10,50mg/L组分别加入对应浓度癌痛克(购自河南省肿瘤研究所,由金蝎、土元、九香虫、大黄、人参、灵芝、黄芪等纯中药组成的粉状制剂,功能:消癌肿、消癌痛、消积水、升白排毒),对照组不加药物,每组设4个复孔。MTT法检测各组细胞24,48,72h增殖率,细胞增殖率=[(A570癌痛克组-A570对照组)/A570对照组]×100%;以流式细胞术检测各组细胞24,48,72h凋亡率;RT-PCR方法检测各组细胞24,48,72h细胞内Rb基因mRNA表达量。结果:①2～50mg/L癌痛克具有明显的增殖抑制作用,癌痛克2,10,50mg/L组在24,48,72h与对照组比较,差异有显著性意义(P<0.05);3组各时点两两之间比较差异有显著性意义(P<0.05);同组各时点之间比较差异有显著性意义(P<0.05)。②2～50mg/L癌痛克组可诱导HepG2细胞凋亡,相同作用时间癌痛克2,10,50mg/L组与对照组比较差异有显著性意义(P<0.001);同一时点各组比较差异有显著性意义(P<0.05);同组不同作用时间点比较差异有显著性意义(P<0.001)。③2～50mg/L癌痛克可上调Rb基因的表达,各浓度组在各时点与对照组比较,差异均有显著性意义(P<0.05);24,48,72h癌痛克2,10,50mg/L组之间比较差异无显著性意义(P>0.05);3组各时点比较差异有显著性意义(P<0.05)。结论:中药癌痛克可通过抑制HepG2细胞增殖或诱导其凋亡,而发挥抗肝癌作用;且其诱导细胞凋亡的机制之一可能为通过增加Rb基因的表达而实现。     

中药天年饮对衰老大鼠脾淋巴细胞增殖转化能力的影响

目的:观察中药天年饮对衰老大鼠抗氧化能力、脾淋巴细胞增殖转化能力及免疫器官指数的影响。方法:实验于2006-04/07在承德医学院基础医学研究所完成。选择清洁级4月龄雄性SD大鼠40只,按随机数字表法分为4组:正常对照组、衰老模型组、天年饮用药组和阴性对照组,每组10只。正常对照组大鼠常规饲养,自由摄食、饮水。衰老模型组、天年饮用药组和阴性对照组大鼠均采用D-半乳糖连续腹腔注射制备亚急性衰老的大鼠模型。衰老模型组大鼠造模成功后不再做任何处理;天年饮用药组大鼠造模成功后每天用天年饮(由炙何首乌、怀牛膝、肉从蓉、淫羊藿、丹参、茯苓等6味中药组成,经熬制后含生药1.5g/mL,购自承德医学院附属肛肠医院)灌胃(8.1g/kg),共30d;阴性对照组大鼠在造模成功后每天用同等剂量的生理盐水灌胃,时间同天年饮用药组大鼠。造模后30d采用噻唑蓝法检测脾淋巴细胞的增殖转化能力,同时分别检测大鼠血清超氧化物歧化酶活性、丙二醛含量以及大鼠的胸腺指数和脾脏指数。结果:纳入动物40只,均进入结果分析。①衰老模型组大鼠血清超氧化物歧化酶活性低于正常对照组、丙二醛含量高于正常对照组,差异有显著性意义[分别为(1.77±0.16),(2.48±0.22)mkat/L,t=2.613,P<0.05;(447.70±31.20),(267.54±40.82)nmol/L,t=11.088,P<0.01];天年饮用药组大鼠血清超氧化物歧化酶活性高于衰老模型组、丙二醛含量低于衰老模型组,差异有显著性意义[分别为(2.33±0.26),(1.77±0.16)mkat/L,t=-2.258,P<0.05;(276.32±23.30),(447.70±31.20)nmol/L,t=13.917,P<0.01]。②衰老模型组大鼠胸腺指数、脾脏指数和脾淋巴细胞的增殖转化能力(A)低于正常对照组,差异有显著性意义(分别为0.0775±0.0309,0.1239±0.0436,t=2.745,P<0.05;0.347±0.099,0.466±0.108,t=2.568,P<0.05;0.388±0.054,0.470±0.063,t=3.125,P<0.01);天年饮用药组大鼠胸腺指数、脾脏指数和脾淋巴细胞的增殖转化能力(A)高于衰老模型组,差异有显著性意义(分别为0.1130±0.0456,0.0775±0.0309,t=2.238,P<0.05;0.451±0.111,0.347±0.099,t=2.211,P<0.05;0.447±0.061,0.388±0.054,t=2.2902,P<0.05)。结论:中药天年饮可提高机体的抗氧化能力及免疫功能,具有延缓衰老的作用。     

针刺足三里和悬钟治疗缺血性脑卒中

目的:观察针刺足三里、悬钟2穴对缺血性脑卒中脑血管功能的影响,分析其可能的作用机制,并对临床疗效做出评价。方法:选择2004-11/2006-05湖北中医药高等专科学校附属古城医院针灸科、荆州市第五人民医院中医康复科、荆州市第三人民医院中医科3单位缺血性脑卒中患者合适病例160例,采用查随机数字表的方法,将其随机分为对照组和针刺组,各80例。对照组采用现代医学常规干预方法进行治疗:卧床,保持呼吸道通畅,预防感染,控制颅内压、血压,维持水电解质平衡。针刺组在此基础上加针刺足三里、悬钟2穴,采用慢速捻转进针法针刺,留针20～30min,每隔5min行针1次。1次/d。两组患者治疗30d。并以经颅多普勒检测观察缺血性脑卒中患者治疗前后脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能的变化,同时以治疗前后神经功能缺损程度为指标评价其临床疗效。结果:160例病例全部进入结果分析。①针刺组与治疗前相比,脑血管舒缩反应能力明显加强,差异有显著性意义(t=2.97,P<0.05),且优于对照组(t=2.45,P<0.05)。②针刺组与治疗前相比,脑血流自动调节能力明显改善,差异有非常显著性意义(t=8.01,P<0.01),且优于对照组(t=7.67,P<0.05)。③针刺组与治疗前相比,大脑半球侧枝循环代偿功能得到加强,差异有显著性意义(t=3.15,P<0.05),且优于对照组(t=5.16,P<0.05)。④针刺组与治疗前相比,神经功能缺损积分明显降低,差异有非常显著性意义(t=4.83,P<0.01),且优于对照组(t=5.43,P<0.05)。结论:针刺足三里、悬钟2穴对缺血性脑卒中患者脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能有明显改善作用,并能促进神经功能的恢复。     

再灌注损伤救援激酶信号转导通路在后处理抗再灌注心肌凋亡中的作用

目的:目前对后处理保护机制的研究还较孤立,缺乏上下游调节机制的联系.通过建立在体大鼠心肌缺血再灌注模型,探讨后处理在整体实验下的抗凋亡效应,以及这种心肌保护作用是否通过再灌注损伤救援激酶(reperfusion injury salvage kinase,RISK)信号转导通路来实现.方法:实验于2006-07/2007-05在首都医科大学病理生理学教研室完成.健康成年雄性SD大鼠32只,均分至以下4组:①心肌缺血再灌注组:结扎冠状动脉左前降支(缺血)45 min,再灌注4 h.②后处理组:在再灌注前,给予10s再灌、10s缺血,连续3个循环.③PI3K抑制剂组:在再灌注前5min经颈静脉注射磷脂酰激醇3激酶(PI3K)抑制剂LY-294002(0.3 mg/kg),余同③组.④MAPKK抑制剂组:在再灌注前5 min经颈静脉注射丝裂原激活蛋白激酶激酶(MAPKK)抑制剂PD 98059(0.3 mg/kg),余同②组.再灌注结束后取左室缺血区,使用Western blotting法检测各组心肌组织中凋亡指标Bax,Bcl-2,Caspase-3表达量变化及RISK通路中Akt,P-ERK 1/2及eNOS表达量变化.结果:32只大鼠均进入结果分析.与心肌缺血再灌注组相比,后处理组心肌凋亡显著减轻,Akt、P-ERK 1/2及eNOS含量显著升高;PI3K抑制剂组心肌凋亡较后处理组严重,Akt、eNOS含量降低;MAPKK抑制剂组心肌凋亡较后处理组严重,ERK 1/2含量降低.结论:后处理对再灌注心肌存在明显的抗凋亡作用,该作用可能与RISK信号转导通路的激活有关.     

Electronic verification of donor-recipient compatibility: the computer crossmatch

Background: This article describes standard operating procedures (SOPs) for a computer crossmatch to replace the immediate-spin crossmatch for ABO incompatibility between patient blood samples submitted for pretransfusion testing and the blood component selected for transfusion. These SOPs were developed following recent changes to the Standards for Blood Banks and Transfusion Services of the American Association of Blood Banks (AABB). Study Design and Methods: SOPs were developed, utilizing currently available software, for pretransfusion testing. The SOP for donor unit processing entails bar code entry of the unit number, component name, and ABO/Rh type; computer entry and interpretation of serologic reactions; warning of discrepancies between bar code-entered blood type and result interpretation; and quarantine of the donor unit in such instances. The SOP for patient sample testing requires bar code entry of specimen accession number, which accesses patient demographics; computer entry and interpretation of ABO/Rh tests; repeat blood typing at the time of crossmatch if only one patient blood type is on record; and warning if there are nonconcordant current and historical blood types. The computer crossmatch SOP requires bar code entry of specimen accession and donor unit numbers; release of group O red cells pending resolution of discrepancies; and immediate-spin crossmatch during computer downtime. Tables validated on- site prompt warning messages and prevent both computer crossmatch and release if blood components of the wrong ABO type are selected. Results: These SOPs meet the requirements of the 15th edition of the AABB Standards. Projected annual time savings at this institution are > 100,000 workload recording units. Further benefits include reduced patient sample volume requirements, less handling of biohazardous material, and elimination of unwanted positive or negative reactions associated with the immediate-spin crossmatch. Release of incompatible blood components when the wrong patient blood type is on record is addressed by requiring the use of group O red cells in the absence of two concordant blood types, one of which must be from a current sample. Conclusion: A combination of existing computer programs and carefully developed SOPs can provide a safe and efficient means of detecting donor-recipient incompatibility without performance of serologic crossmatch.     

研究生一年级学生运动等级与其健康状况的相关性

目的:调查研究生一年级学生的体育锻炼和健康状况,并分析其相关性。方法:调查于2005-09/10完成。选取西南大学和重庆大学2005级研究生一年级学生作为调查对象,共发放问卷190份,男90份,女100份。进行问卷调查,当场收回问卷。问卷内容:①自测健康评定量表:由48个条目构成,1～17为自测生理健康评定子量表(170～0分),19～34为自测心理健康子量表(150～0分),35～47为自测社会健康子量表(120～0分),分数越高代表越健康。②体育活动等级量表:主要调查调查对象上1个月参加体育锻炼的运动量,运动量=强度×时间×频率,强度与频率从1～5等级分别计1～5分,时间从1～5等级分别计0～4分,故运动量最高分为100分,最低分为0分。结果:有效回收176份问卷进入结果分析。①不同性别调查对象在运动等级上的分布情况:研究生一年级男女新生的运动量水平都偏低(男生:31.80±23.19,女生:15.57±15.39),且性别对运动量有显著的主效应(P<0.01),女生在运动量上的得分显著偏低。②调查对象健康状况各维度与运动量的相关性:运动量与总体健康、生理健康、心理健康和社会健康都存在显著相关性(r=0.150～0.238,P<0.05～0.01),生理健康对运动量有显著的预测作用(P<0.05)。③不同运动量等级调查对象的健康量表得分:运动等级对生理健康有显著的主效应(P<0.01),随着运动量的增加,研究生一年级学生的生理健康水平不断提高。结论:研究生一年级学生的体育锻炼运动量与健康水平存在着明显的正相关,随着运动量的增加,生理健康、心理健康及社会健康水平均不断增加。     

Placebo Analgesia: Clinical considerations

Pain is the predominant symptom that prompts patients to seek medical advice and treatment from physiotherapists. Various treatment modalities such as heat and cold, electrical stimulation (Cheing and Hui-Chan, 1999), ultrasound, manipulative techniques, massage and laser treatment have been demonstrated in varying degrees to be clinically effective for managing pain of different pathologies. However, all these treatments could be assumed to have some placebo elements (French, 1994).

From a research design perspective, the presence of placebo response is undesirable and must be controlled as it complicates the demonstration of ‘real' treatment effect. From a clinical perspective, it is intriguing to note that the condition of patients in the placebo control groups did improve considerably in many of these validation studies, although in the majority the improvement was not so marked as in the treatment groups. Conspicuously, some neuro-physiological and psychological aspects of the placebo effects may have clinical use in enhancing the effect of pain treatments and their outcomes.

Unfortunately, although placebo response has been a subject of continuing interest among some physiotherapy researchers and clinicians, information about placebo analgesia and its clinical utility is seldom discussed. The purpose of this paper is to provide clinicians with an overview of the construct and research related to placebo analgesia as well as a discussion of the potential clinical use of certain components of placebo analgesia to enhance pain rehabilitation outcomes in physiotherapy practice.     

Moderate and severe reactions in blood donors

During the period April 1985 to March 1986, 217 blood donors were found to have moderate (syncopal) to severe (convulsive) reactions. This population was compared to 5630 randomly selected donors who did not have reactions. An examination of demographic, physical, and societal/emotional factors was conducted to determine if any were predictive of reactions in donors. The results of the research supported the hypothesis that first-time donors have a higher frequency of reactions (1.7%) than do repeat donors (0.19%). A review of the above predictive factors documented that, with regard to demographic factors, 1) the number of prior donations was inversely proportional to the risk of reaction; 2) the gender of the donor was not predictive; and 3) youth was a predictor of reactions. An analysis of the physical factors revealed that donors who reacted were of lower weight (mean, 153.7 lb) than those who did not (mean, 166.4 lb) and that systolic blood pressure was slightly lower in the group with reactions. Although the difference was significant (3 torr), it was not thought to be significant clinically. In a comparison of a group with systolic blood pressure ranging from 80 to 100 torr and a group with systolic blood pressure ranging from 120 to 140 torr, the first group had a 70-percent higher risk of reaction. Finally, with regard to the last category of societal or emotional factors, the research demonstrates 1) that the ingestion of caffeinated beverages was associated with a reduced risk of reactions; 2) that the food intake of donors who reacred was significantly different from that of those who had no reaction, but this difference was not thought to be clinically significant; and 3) that the duration between registration and the onset of phlebotomy was directly predictive of reaction status. The research indicates that first-time donor status and several specific demographic, physical, and societal or emotional factors are predictors of donor reactions.