Abnormal processing of the nociceptive input in Parkinson's disease: a study with CO2 laser evoked potentials

Since a number of patients with Parkinson’s Disease (PD) complain of painful sensations, we studied whether the central processing of nociceptive inputs is abnormal in PD. To test this hypothesis, we recorded scalp CO₂ laser evoked potentials (LEPs) to hand skin stimulation in 18 pain-free PD patients with unilateral bradykinetic-rigid syndrome (hemiparkinson) during the off state and in 18 healthy subjects. This technique allows us to explore non-invasively the functional status of some cerebral structures involved in nociceptive input processing. In both PD patients and control subjects, CO₂ laser stimulation gave rise to a main negative N2 potential followed by a positive P2 response at vertex peaking at a latency of about 200 and 300 ms, respectively. These potentials are thought to originate from several brain structures devoted to nociceptive input processing, including the cingulate gyrus and insula. PD patients and normal subjects showed comparable N2 and P2 latencies, whereas the N2/P2 peak-to-peak amplitude was significantly lower in PD patients (regardless of the clinically affected body side) than in controls. LEPs were even recorded after acute L-dopa administration in 7 additional PD patients. L-dopa administration yielded no significant change in N2/P2 amplitude as compared to the off state. These results suggest an abnormal nociceptive input processing in pain-free PD patients which appears to be independent of clinical expression of parkinsonian motor signs and is not affected by dopaminergic stimulation.     

Reducing Diagnostic Errors in Musculoskeletal Trauma by Reviewing Non-Admission Orthopaedic Referrals in the Next-Day Trauma Meeting

INTRODUCTION

Diagnostic errors in orthopaedics are usually caused by missing a fracture or misreading radiographs. The aim of this study was to document the pick-up rate of the wrong diagnoses by reviewing X-rays and casualty notes in the next-day trauma meeting.

PATIENTS AND METHODS

The casualty notes and radiographs of 503 patients were prospectively reviewed in the daily trauma meeting between August 2002 and December 2002 in a district general hospital. The relevant data were collected and analysed by a single assessor.

RESULTS

The false positive rate for making an orthopaedic diagnosis was 12.6% (i.e.) diagnosing a fracture, when none existed). The false negative (missing) rate was 4%, while 2.4% incidental findings were missed, or at least not documented, after reading the X-rays. There were 7.8% wrong diagnoses made. The majority of the patients were seen by the senior house officers.

CONCLUSIONS

The medicolegal significance of false negative diagnosis is obviously greater. In a busy emergency department, where a large number of patients are seen, there is a greater risk. This study shows the importance in a small-to-medium sized accident and emergency unit as well, where there is no senior cover available out-of-hours for final radiological interpretation. A morning trauma meeting which covers reviewing admitted patients as well as non-admission orthopaedic referrals has an effective risk management solution to early detection of missed and wrong diagnoses.     

Boosting of SIV-specific T cell responses in rhesus macaques that resist repeated intravaginal challenge with SIVmac251

Despite repeated high-risk exposure to infectious HIV-1, some individuals remain HIV-1 seronegative and apparently uninfected. The use of nonhuman primate model systems to study SIVmac transmission may help to elucidate the factors responsible for protection in exposed, seronegative (ESN) humans. In an earlier vaccination study, three control rhesus macaques that were exposed to three sequential intravaginal challenges with pathogenic SIVmac251 failed to show evidence of infection after 5 years of observation. 51Cr release assay results suggested that these animals had low-level cytotoxic T lymphocyte responses to SIVmac proteins. We hypothesized that these responses might be an important component of protection from mucosal challenge. We performed an additional intravaginal challenge of all three macaques and monitored SIV-specific T cell responses in peripheral blood, using the sensitive enzyme-linked immunospot (ELISpot) assay. After the fourth challenge, one animal became infected; this animal did not mount a strong SIV-specific T cell response. Two other macaques remained uninfected as determined by peripheral blood mononuclear cell (PBMC) coculture, polymerase chain reaction (PCR), and branched DNA (bDNA) analysis of peripheral blood and lymphoid tissues, but demonstrated boosting of SIV-specific T cell responses after challenge. These results support a protective role for SIVmac-specific T cells in repeatedly exposed, persistently seronegative rhesus macaques.     

Development and test of a new plantar pressure control device for foot unloading]

OBJECTIVE: To validate a new plantar pressure control device able to detect excessive plantar pressure on-line and to improve the subject's awareness of the gait pattern in order to correct and optimize the load distribution patterns. The aim of this study was to verify the feasibility of a deliberate and partial unload of the first metatarsal head. MATERIAL AND METHOD: Eight healthy subjects were equipped with the plantar pressure sensors integrated in customized insoles. They were told to unload the first metatarsal head of the right foot by 5%. During the trials, the auditory and visual signals inform the subject of an excessive, insufficient or correct unload. RESULTS: Five subjects over eight succeeding in modifying significantly their gait pattern and in unloading by 5% the first metatarsal head. The unload is effective in 70.7% of the trials. However, the subjects were spontaneously inclined to reduce peak pressure under the first metatarsal head beyond the critical peak plantar pressure threshold (48.2% of the trials). DISCUSSION - CONCLUSION: The results showed the feasibility of a partial and deliberate foot unloading in using biofeedback device but emphasized the subject's difficulty to control a discriminated and accurate unload of the first metatarsal head. That points out the necessity of learning period. The baropodometric biofeedback rehabilitation focuses on the subject's capacities to modify its own locomotor pattern. It would be used as primary and secondary prevention means of diabetic foot ulceration.     

应用功能磁共振成像分析脑卒中患者康复治疗前后大脑再塑机制

目的:应用功能磁共振成像观察脑卒中后及康复过程中,在相应脑内运动功能区激活的变化情况,探讨不同运动模式下皮质功能再塑的表现。方法:选取2003-02/10大庆油田总医院康复科住院的皮质下脑梗死患者8例,在发病后1周始进行连续两个月的康复。在康复前、康复1,2个月时运用Brunnstrom分级、Caroll上肢功能量表(0￣100分,评分越高功能越好)对其手功能进行评价,并采用GEMR/iHiSpeed1.5超导磁共振扫描机进行磁共振成像功能激发检查。患者用病手执行简单运动(快速连续的拇指与其他各指的对指动作)、随意运动(用病手摸不同形状的木块),获得脑功能激发图像,观察脑内相关功能区的激活情况。结果:8例受试者均进入结果分析。①康复后所有患者Brunnstrom分级和Caroll上肢功能评分均较康复前有明显改善。②病手简单运动时脑内相关功能区的激活情况:8例受试者7例在损伤后早期手指不能对指,所以没有激活;M1,SMA,PMA脑区和小脑呈现单侧激活-双侧激活-单侧激活的变化过程;随着运动功能恢复,脑内激活数目随时间呈下降趋势,几乎接近正常人脑功能表现。③病手随意运动时脑内相关功能区的激活情况:实验中发现引起的运动相关功能区的激发情况变化多样,规律性较差,但其中5例受试者表现出损伤后激发数目明显减少,许多对运动起决定性支配作用的功能区亦不激活;随着运动功能恢复,激发区数目呈上升趋势,同损伤后简单运动的激活表现。结论:①脑卒中后病手经过康复治疗简单运动恢复较好,康复治疗2个月后脑内运动功能相关区域激活的规律已同正常人。②脑卒中后病手随意运动恢复较困难,康复治疗后不如简单运动恢复好,脑内相关运动功能区激活无明显的规律性。随着运动功能的恢复,脑内相应的运动功能区激活增多。     

原发性血小板减少性紫癜模型小鼠血浆内皮素水平与加味小柴胡颗粒及其组方的干预

目的:建立原发性血小板减少性紫癜动物模型,观察加味小柴胡颗粒及其组方对造模小鼠血浆内皮素含量的调节。方法:实验于2006-07/12在南京中医药大学第一临床医学院实验中心完成。①实验动物与药品:选取SPF级BALB/C小鼠54只,随机数字表法分为5组:正常组10只、模型组12只、加味小柴胡颗粒组11只、小柴胡颗粒组11只、丹参三七颗粒组10只。小柴胡颗粒主要成份为柴胡、黄芩、法半夏、党参、炙甘草、生姜、大枣,每克颗粒含生药5.45g;丹参三七颗粒每克含生药2g;加味小柴胡颗粒:在小柴胡颗粒主要成份上添加丹参、三七,每克颗粒含生药4.18g;均由天江制药厂提供。②实验方法:除正常组外,各组均建立原发性血小板减少性紫癜动物模型。造模后7d灌胃给药,加味小柴胡颗粒组给予加味小柴胡颗粒16.7g生药/kg体质量,小柴胡颗粒组给予小柴胡颗粒13.8g生药/kg体质量,丹参三七颗粒组给予丹参三七颗粒4.4g生药/kg体质量,正常组及模型组均给予等容积蒸馏水。每天给药1次,共给药14d。③实验评估:各组小鼠眼球取血,分离血浆,取上清液用放射免疫法测定血浆内皮素含量。结果:54只BALB/C小鼠均进入结果分析。与正常组比较,模型组血浆内皮素含量明显升高(P<0.05)。与模型组比较,加味小柴胡颗粒组、丹参三七颗粒组明显下降(P<0.05),而小柴胡颗粒组无明显变化(P>0.05)。结论:原发性血小板减少性紫癜血管内皮功能异常,表现为血浆内皮素明显升高,加味小柴胡颗粒及丹参三七颗粒均对其有下调作用,而小柴胡颗粒无影响。     

富血小板血浆对人骨髓间充质干细胞成骨分化的抑制效应

目的:一些理论质疑富血小板血浆对骨前体细胞成骨分化的作用,本实验拟验证富血小板血浆对体外培养的人骨髓间充质干细胞成骨分化的抑制效应。方法:实验于2005-05/11在南方医科大学组织工程试验室(省级)完成。①实验方法:抽取6名健康志愿者髂前上棘骨髓5mL进行体外细胞培养扩增,静脉血10mL以二次离心法制得富血小板血浆。诱导骨髓间充质干细胞时富血小板血浆与骨髓间充质干细胞均来自同一个体。②碱性磷酸酶染色:取第4代骨髓间充质干细胞,分为两组:富血小板血浆组加入富血小板血浆使终浓度为100g/L,单纯血清培养组仅加入等量胎牛血清。培养后第7天进行碱性磷酸酶染色,阳性细胞为胞质中呈现黑色颗粒或块状沉淀。③矿化结节染色:取第4代骨髓间充质干细胞,分组同上。培养后第19天以0.1%茜素红-TrisHcl(pH8.3)37℃下放置30min,矿盐沉积染色阳性为红色。④Cbfa1基因表达:取第4代骨髓间充质干细胞,分组同上。培养后第3,7,12,16天RT-PCR法检测骨髓间充质干细胞Cbfa1基因的表达。⑤形态学观察:实验过程中使用相差显微镜观察各组细胞生长情况及形态学变化。结果:①骨髓间充质干细胞碱性磷酸酶染色结果:培养后第7天,富血小板血浆组碱性磷酸酶阳性细胞数量较单纯血清培养组明显减少,且阳性细胞内灰黑色颗粒也明显减少,为弱阳性。②骨髓间充质干细胞矿化结节染色结果:培养后第19天,单纯血清培养组可见细胞表面有较多的矿盐沉积,但未形成明显的矿化结节。富血小板血浆组细胞表面只有稀少的矿盐沉积。③骨髓间充质干细胞cbfa1mRNA的表达:培养后第3,7,12,16天,随着培养时间的延长单纯血清培养组与富血小板血浆组cbfa1基因表达量均逐渐增高,同一时间点两组间cbfa1基因的表达基本相似。④骨髓间充质干细胞形态学变化:富血小板血浆组骨髓间充质干细胞增殖旺盛,细胞达到单层汇合的时间较单纯血清培养组明显缩短。单纯血清培养组细胞在完全汇合后开始出现聚合现象(14～16d),但趋向性不明显,未完全形成团簇;富血小板血浆组细胞在完全汇合后未出现聚合现象,细胞密集生长。培养初期两组细胞以梭形为主,多角形细胞较少,培养至14～16d单纯血清培养组多角形细胞较富血小板血浆组增多。结论:富血小板血浆可抑制人骨髓间充质干细胞碱性磷酸酶的分泌与矿盐沉积,对人骨髓间充质干细胞成骨分化的直接效应是抑制其分化。     

大网膜脂肪组织中胰岛素受体底物1、葡萄糖转运蛋白4和抵抗素mRNA表达水平与代谢综合征的关系

目的:分析代谢综合征患者大网膜脂肪组织胰岛素受体底物1、葡萄糖转运蛋白4和抵抗素mRNA表达水平及其与代谢综合征相关指标的关系。方法:选择2003-02/08在青岛大学医学院附属医院普通外科及妇科择期手术的患者53例,均知情同意。根据代谢综合征的诊断标准分为两组:①代谢综合征组28例,分为两个亚组:2型糖尿病组13例,非糖尿病组15例。②对照组25例。手术前当天抽取患者空腹血,测定空腹血糖、三酰甘油、空腹胰岛素。手术中取大网膜脂肪组织约200mg,用于RNA提取。采用一步法半定量反转录聚合酶链反应技术,测定患者大网膜脂肪组织中胰岛素受体底物1、葡萄糖转运蛋白4以及抵抗素的mRNA表达水平。结果:代谢综合征组28例,对照组25例患者全部进入结果分析,无脱落。①代谢综合征组患者大网膜脂肪组织胰岛素受体底物1、葡萄糖转运蛋白4mRNA表达均显著低于对照组(P<0.01)。2型糖尿病患者的胰岛素受体底物1mRNA、葡萄糖转运蛋白4mRNA表达显著低于非糖尿病患者(P<0.01)。②代谢综合征组与对照组患者胰岛素受体底物1与葡萄糖转运蛋白4的mRNA表达呈显著正相关(r=0.661,0.621,P<0.01)。③多因素逐步回归分析显示腰臀比与胰岛素抵抗指数及胰岛素受体底物1、葡萄糖转运蛋白4mRNA表达均有明显的相关性。④抵抗素mRNA表达阳性率和表达量在2型糖尿病代谢综合征患者、非糖尿病代谢综合征患者与对照组之间差异均无显著性意义(P=0.121,P=0.228),与腰臀比、体质量指数、胰岛素抵抗指数、空腹血糖、空腹胰岛素、三酰甘油以及血压均无相关性(P>0.05)。结论:代谢综合征患者大网膜脂肪组织胰岛素受体底物1与葡萄糖转运蛋白4mRNA表达明显降低,其中以2型糖尿病患者表现最为显著。大网膜脂肪组织抵抗素mRNA表达与代谢综合征及2型糖尿病无关。胰岛素受体底物1、葡萄糖转运蛋白4的mRNA表达和腰臀比可联合预测胰岛素抵抗的程度。     

人脂肪干细胞复合脱细胞软骨基质支架在生物反应器中构建组织工程软骨

目的:观察人脂肪干细胞复合脱细胞软骨基质支架在生物反应器中初步构建组织工程软骨的可行性。方法:实验于2005-04/2006-05在解放军总医院骨科研究所完成。脂肪组织和关节软骨均来自膝关节置换术中切除的组织,并经患者知情同意。关节软骨冻干后经粉碎机粉碎,过筛,选取25～38μm大小的软骨微粒。在样品中先加入2.5g/L胰蛋白酶,37℃消化24h,再加入1%Triton X-100震荡72h。将软骨微粒和蒸馏水按1∶3的比例混合后滴加在模板中,置入冷冻干燥机冻干后行紫外线交联。紫外线照射8h完成。最后经25kGy 60Co辐照灭菌完成支架制备。取膝关节置换术中切除的髌下脂肪垫,酶消法获得脂肪干细胞,扩增后复合于脱细胞软骨基质制成圆柱状三维支架上(细胞密度5×1010L-1),置于生物反应器中进行诱导培养,同时设静态培养组作为对照,3周后观测大体形态和组织学形态变化,同时进行组织化学(包括番红花O,阿利新蓝染色)和Ⅱ型胶原免疫组织化学分析。结果:生物反应器组诱导培养3周苏木精-伊红染色显示支架结构消失,只有中心区域残存少量支架结构;静态培养组支架结构尚存在,有少量基质分泌。番红花O染色显示生物反应器组细胞外有大量蛋白聚糖沉积,阿利新蓝染色表明有软骨特异性蛋白多糖的聚集;而静态培养组只有部分区域染色且淡于生物反应器组。Ⅰ型胶原免疫组化的结果显示,在生物反应器组细胞能够合成大量软骨细胞特异性胶原成分,而静态培养组呈弱阳性。结论:生物反应器培养明显促进了脂肪干细胞的增殖与软骨分化,是体外构建组织工程软骨的良好方法。