干细胞移植改善股骨头坏死缺血状态的实验及临床效应：20只模型兔及188例患者资料分析

背景：缺血性股骨头坏死由于股骨头局部滋养血管损伤、供血不足致病，迄今尚无最佳治疗方案，改善股骨头血液供应成为治疗本病的关键。 目的：通过动物实验和临床观察，验证自体骨髓问充质干细胞移植血管再生技术改善股骨头坏死缺血状态的效果。 设计：对比动物实验和自身对照临床观察。 单位：解放军第四六三医院细胞治疗中心。 资料：①动物：选用日本大耳白兔20只，雌雄不拘，体质量3．0—4．0kg，购于沈阳军区总医院动物实验中心。此动物实验符合动物伦理学要求。②对象：选择2004—07／2007—07在解放军第四六三医院细胞治疗中心住院的具有完整随访资料的缺血性股骨头坏死患者188例（335髋），男113例，女75例，年龄18-72岁。均经髋关节X线摄片、核素扫描、MRI、CT检查确诊；对治疗和实验知情同意。实验经医院伦理委员会批准。 方法：①动物实验：实验于2004—01／2004—06在沈阳军区总医院动物实验中心完成。采用液氮冷冻股骨头缺血性坏死模型建立方法，双后肢股骨头均制作缺血模型，右侧为移植组，左侧为对照组。抽取骨髓，将所获单个核细胞经右股动脉注入；同法将生理盐水注入左侧股动脉内。②临床观察：密度梯度离心缺血性股骨头坏死患者自体骨髓，分离单个核细胞。行旋股内、旋股外及闭孔动脉干细胞移植术。 主要观察指标：①4周后行DSA动脉造影观察双侧血管新生情况。4，12周后取双侧股骨头标本，行病理切片观察新骨再生及股骨头修复程度。②于移植后第3，6，12，24个月观察患者髋关节疼痛、行走间距及步态、髋关节外展与内旋功能变化。于干细胞移植后6个月行股骨头供血动脉造影术，观察血管新生及股骨头供血动脉充盈情况。于干细胞移植后6，12，24个月行CT，X射线，MRI检查，观察股骨?     

经动脉骨髓干细胞移植治疗股骨头坏死63例

目的：观察经动脉骨髓干细胞移植治疗股骨头坏死的效果。 方法：选择2004—08／2005—12在解放军第四六三医院细胞治疗中心收治的股骨头坏死接受动脉内骨髓干细胞移植治疗患者63例113髋。年龄12-54岁（平均33岁），曾经应用皮质激素史29例，外伤史17例，大量饮酒史11例，原因不明6例。63例患者均行髋关节X射线摄片检查确诊，15例同时行髋关节MRI检查，38例行髋关节CT检查。根据Fieat分期法Ⅱ期52髋（46.0％），Ⅲ期58髋（51.3％），Ⅵ期3髋（2，7％）。①采集自体骨髓200-400mL，Ficoll密度梯度离心，分离单个核细胞（1～4）&;#215;10^11L^-1，CD34^＋细胞1．4％～3．5％（2．4％），CD133^＋细胞1．13％～3．25％（1．88％），制备成单个核细胞悬液10-20mL。②在数字减影血管造影术下行股动脉穿刺，将导管超选插入旋股内动脉、旋股外动脉及闭孔动脉，将细胞悬液缓慢灌注入动脉内。⑧干细胞移植后随访观察患者髋关节疼痛程度、疼痛性质及疼痛时间变化、行走距离及步态变化、髋关节外展与内旋功能变化；6个月后复查股骨头供血动脉造影，观察血管新生及股骨头供血动脉充盈情况；观察干细胞移植后不良反应。12个月后行髋关节X射线摄片、CT，MRI扫描观察股骨头形态学变化。结果：63例患者关节疼痛、行走距离及关节功能平均随访3．2个月，其中5例行数字减影血管造影术，15例行X射线检查（2例同时做CT检查）。①随访时观察髋关节疼痛有不同程度的缓解54例（54／63，85．7％），关节功能改善19例（19／63，30．2％），行走间距延长34例（34／63，54％），生活质量提高。②干细胞移植术后6个月5例患者行数字减影血管造影术股骨头供血动脉造影检查，显示旋股内动脉、旋股外动脉及闭孔动脉管径增粗，新生血管增多，血流速度增快，与移植前血管造影结果比较，股骨头区血液供应明显改善。③2例18个月X射线摄片、CT扫描股骨头坏死区缩小，可见新骨形成。④3例患者在治疗中未发生严重并发症和不良反应。结论：经动脉干细胞移植治疗缺血性股骨头坏死方法简便、安全有效，在治疗中未发生不良反应，是治疗缺血性股骨头坏死的一种新手段。     

全麻下行双侧扁桃体切除术诱发甲状腺功能亢进危象1例

<正>患者,女,21岁。因"扁桃体切除术后苏醒不佳2 h"于2013年1月28日14时转入我院急诊科。既往有睡眠呼吸不畅,入院前1周加重,就诊于我院耳鼻喉科,经检查诊断为"扁桃体肥大",于1月28日上午在全麻下行双侧扁桃体切除术,手术顺利,转入苏醒室待醒。但患者并未按常规正常苏醒,并逐渐出现心率加快,发热,心率在短时间内达170次/min,体温很快升到39.2℃。急请心内科、呼吸科、内分泌科等多科会诊,发现患者呈轻度突眼症,术前心电图提示心率110次/min,因患者呈昏迷状态,且体型偏胖,甲状腺触诊不清,急检查甲状腺功能,结果:血TT38.0 ng/mL,TT426.6μg/dL,FT3     

自体骨髓单个核细胞经介入途径移植治疗股骨头坏死54例：12个月疗效随访

目的:观察经介入途径移植自体骨髓单个核细胞在股骨头坏死治疗中的应用,并评价其疗效。方法:选择2004-07/2005-11在解放军四六三院细胞治疗中心住院的,具有完整随访资料的股骨头坏死确诊患者共54例91髋。纳入确诊股骨头坏死,有关节疼痛、功能障碍等症状患者,性别、年龄不限;排除有严重心力衰竭、严重肾功能异常等不能耐受手术者。符合纳入标准54例,男45例,女9例,12～68岁。按ARCO分期Ⅱ期42髋,Ⅲ期47髋,Ⅳ期2髋。实验对象对治疗的相关内容知情同意并签知情同意。干预措施:抽取患者髂后上嵴骨髓进行单个核细胞悬液的制备。在DSA监视下将采集的单个核细胞混悬液经股动脉行Seldinger法穿刺,穿刺成功后,置入4F动脉鞘,经动脉鞘置入Cobra导管,将导管超选择至闭孔动脉及旋股内外侧动脉,平均注入单个核细胞悬液。术后定期随访症状变化情况,1年后复查X射线或CT,随访疼痛、关节活动度等情况。实验评估:①疼痛指数:无疼痛症状为3分,Harris髋关节评分疼痛分级A级;时有隐痛2分,Harris髋关节评分疼痛B级;轻度疼痛为1分,Harris髋关节评分疼痛C级;中度疼痛为0分,Harris髋关节评分疼痛D级。②功能指数:髋关节屈、伸、展、收、旋转度评分达Harris髋关节活动范围评分4～5分为3分;3～4分为2分;2～3分为1分;小于2分为0分。③X射线平片指数:股骨头形态无变化,应力骨小梁清晰,坏死区明显缩小为3分;坏死区略缩小为2分;治疗前后无明显变化为1分;坏死区扩大为0分。④血管指数:治疗后旋股内、外侧动脉及其分支增粗、增多,延长1cm以上者3分;1～0.5cm者2分;小于0.5cm者1分,无变化者0分。结果:54例患者均完成疼痛症状、关节功能及影像学随访1年。①术后12个月复查疼痛消失9髋,缓解61髋,无缓解21髋,缓解率为76.9%。②关节功能缓解33髋,无缓解58髋,缓解率为36.3%。③1年后X射线平片或CT、MRI示股骨头区可见不同程度的股骨头坏死区骨质密度改变,坏死区有吸收、缩小,股骨头形态变圆滑规整,改善28髋,无缓解或加重63髋,缓解率为30.1%。④12例24髋完成术后12个月复查股骨头供血动脉数字减影血管造影,好转18髋,好转率为72.2%。结论:经介入途径移植自体骨髓单个核细胞治疗股骨头坏死损伤小,可缓解临床相关症状。     

超选择性动脉干细胞移植治疗创伤性股骨头缺血性坏死的研究

目的研究自体骨髓干细胞（BMsC）移植对创伤性股骨头坏死患者缺血状态的改善程度和治疗效果。方法选取2004年8月-2008年12月期间127例139髋成人创伤性股骨头坏死患者,应用自体BMSC移植治疗。治疗方法：分别采集127例患者骨髓各200～300ml,采用Ficoll密度梯度离心法分离单个核细胞;流式细胞仪测定CD34＋和CD133＋在所分离出的干细胞悬液中的含量。将单个核细胞用生理盐水制备成悬液10～20ml。采用数字减影血管造影技术（DSA）行超选择性股骨头供血动脉内干细胞移植术。按世界骨循环研究学会（ARCO）对骨坏死分期,自身前后对照观察疗效。移植后第3、6、12、24个月,根据髋关节Harris评分评价疗效,移植术后6个月通过复查患者股骨头供血动脉DSA观察其是否有新血管形成,每隔6个月采用影像学方法观察股骨头形态学变化。结果（1）临床疗效：对接受自体BMSC移植治疗的127例患者随访3。24个月（平均18．3个月）,其中髋关节疼痛缓解患者111例（占患者总数的87．6％）,髋关节功能改善患者107例（占患者总数的84．3％）,行走间距延长患者109例（占患者总数的85．9％）;（2）影像学检查：干细胞移植术后6个月,127例患者中随访到24例患者行股骨头供血动脉DSA检查后,显示供血动脉较移植前明显增多、增粗,血流速度增快;12～24个月后36例患者股骨头区骨质病变获得改善。结论超选择性动脉干细胞移植方法简便、安全有效,对因缺血导致坏死的股骨头无再次损伤,能够有效地治疗创伤性缺血股骨头坏死。     

自体外周血干细胞移植治疗糖尿病足26例临床研究

目的　观察自体外周血干细胞移植治疗糖尿病足的临床疗效及相关影响因素。方法　自 2 0 0 3- 112 0 0 4 - 0 7解放军第 4 6 3医院对 2 6例自愿接受自体外周血干细胞移植的糖尿病足患者进行外周血干细胞动员 ,给予rhG -CSF每天 4 5 0 6 0 0 μg ,皮下注射 ,第 6天用COBE 6 1血细胞分离机单采干细胞 ,总量 82 14 8mL。将干细胞悬液按 3cm×3cm距离进行缺血肢体移植术 ,移植后观察各项指标综合评估。结果　移植后 314d 2 3例患者疼痛明显缓解 ,2 7d2 4例患肢冷感有不同程度的改善 ,4 12周 4例溃疡明显好转 ,踝压指数 (ABI) 6例患者改善。 2例DSA下肢动脉造影结果显示新侧支血管形成明显增加。未出现并发症和不良反应。结论　自体外周血干细胞移植治疗糖尿病足是一种简便、安全、有效的治疗方法     

骨髓间充质干细胞移植治疗杜氏型进行性肌营养不良症344例临床疗效分析

目的 探讨自体骨髓间充质干细胞(MSCs)及脐带间充质干细胞移植治疗杜氏型进行性肌营养不良症的临床疗效.方法 2007年6月～2008年6月该科室应用自体MSCs移植对344例杜氏型肌营养不良症患者进行治疗,采用治疗前后对照方法进行研究.骨髓间充质干细胞制备:对344例患者皮下注射粒细胞集落刺激因子,剂量为每日每公斤体重5～10μg,动员骨髓干细胞,共4 d,4 d后采集骨髓,将采集的骨髓通过Percoll梯度离心,提取单个核细胞(MNC),总量为(5.72±1.89)X108,CD34+细胞(2.67±1.22)%,CD133+细胞(1.79±0.73)%.培养骨髓单个核细胞7～10d,最终得到骨髓间充质干细胞含量为(1.90±0.96)X10s.应用四肢肌肉内注射的方式进行干细胞移植.移植后3、6、9、12个月观察患者酶学、肌电图、肌肉MRI变化,评价干细胞移植前后日常生活活动能力(ADL)、临床分级.结果 344例患者中292名患者四肢肌肉力量均有所改善.总有效率为85.00%,好转患者中199名患者四肢力量明显好转,明显好转率为58.00%;93名患者四肢肌力较前略好转,好转率为37.00%.结论 自体骨髓干细胞移植可使杜氏型肌营养不良症患者肌力得到改善,生活质量得到提高.     

干细胞移植改善股骨头坏死缺血状态的实验及临床效应：20只模型兔及188例患者资料分析

BACKGROUND: Ischemic femoral head necrosis is caused by local vascular injury and blood-supply insufficiency. There exists no optimal treatment for the ischemic femoral head necrosis. Thus, the improvement of the blood supply to the femoral head seems to be a key point for the treatment. OBJECTIVE: To verify the curative effects of autologous bone marrow mesenchymal stem cell transplantation induced vascular regeneration on the improvement of ischemic femoral head necrosis via animal experiments and clinical observations. DESIGN: Contrast animal experiment and self-controlled clinical observation. SETTING: Cell Therapy Center, the 463 Hospital of Chinese PLA. MATERIALS AND PARTICIPANTS: ① Animals: Twenty Japanese white rabbits in either gender and weighing 3.0-4.0 kg were purchased from Animal Experimental Center, General Hospital of Shenyang Military Area Command of Chinese PLA. The animal experiments were coincident with the ethical standards. ② Participants: 188 patients with ischemic femoral head necrosis (335 hips) having whole following-up data were selected from Cell Therapy Center, the 463 Hospital of Chinese PLA from July 2004 to July 2007. There were 113 males and 75 females, and their ages ranged from 18 to 72 years. Diagnosis was done by using X-ray photographs, nuclide scanning, MRI and CT examinations. All patients provided the informed consent, and the study was approved by the local research ethics committee. METHODS: ① Animal experiments: The experiment was carried out at the Animal Experiment Center, General Hospital of Shenyang Military Area Command of Chinese PLA from January to June 2004. Ischemic femoral head necrosis models were established at both hindlimbs by using liquid-nitrogen refrigeration. The right side was regarded as transplantation group and the left one as control group. Mononuclear cells extracted from bone marrow were poured in the right femoral artery, while saline was poured in the left femoral artery. ②Clinical observations: Mononuclear cells were separated from autologous bone marrow of patients with ischemic femoral head necrosis after density gradient centrifugation. MAIN OUTCOME MEASURES: ①Four weeks later, angiogenesis at both femoral arteries was observed by arteriography by using digital subtraction anglography (DSA). Moreover, bilateral femoral head samples underwent pathological sections to observe bone regeneration and repair of femoral head 4 and 12 weeks later. ②Items including hip pain, walking distance and gait, abduction and internal rotation function changes of hip joint were observed in 3, 6, 12 and 24 months after stem cell transplantation in media femoral circum flex artery, lateral femoral circum flex artery and obturator artery. In 6 months after stem cell transplantation, angiogenesis and blood supply of femoral head were observed by using arteriography. In 6, 12 and 24 months after stem cell transplantation, morphological and ischemic changes of femoral head were observed by using CT, X-ray and MRI examinations. Harris scores were used to evaluate function of hip joint before and in 3, 6, 12 and 24 months after stem cell transplantation. RESULTS: Animal experiment: Twenty rabbits were involved in the final analysis. ① DSA-arteriography results: In 4 weeks after transplantation, blood-supply arteries in femoral head of right hindlimb in the transplantation group were more than those in the control group. ② Pathological results: In 12 weeks after transplantation, cartilage, lamellar bone and bone trabecula in the left femoral head were repaired remarkably, but left femoral head necrosis was not improved. Clinical observations: 188 patients were involved in the final analysis. ① Improvement of symptoms: Among 188 patients, 164 (87.3%) had remission of hip pain, 147 (78.4%) had function improvement, and 150 (80.0%) had elongation of walking distance. ② Imaging changes: At 6 months after transplantation, DSA-arteriography in 12 patients demonstrated that blood-supply arteries in femoral head were increased and thickened remarkably as compared with those before transplantation, and the blood flow was rapid. At 12-24 months after transplantation, lesion of bone matrix in 24 patients was improved under the X-ray, CT and MRI examinations. ③ Harris scores of hip joint: The scores at 6, 12 and 24 months after transplantation were significantly higher than those before transplantation (t= -3.423, -6.714, -9.039, P < 0.01). CONCLUSION: Autologous bone marrow mesenchymal stem cell transplantation can effectively improve and treat ischemic femoral head necrosis.