大鼠脊髓损伤后不同时期移植人脐带间充质干细胞的修复效果观察

目的:观察大鼠脊髓损伤后不同时期局部移植人脐带间充质干细胞(human umbilical cord mesenchy-mal stem cells,hUCMSCs)修复脊髓损伤的效果,探讨hUCMSCs局部移植治疗脊髓损伤(spianl cord injury,SCI)的最佳移植时机。方法:雌性Wistar大鼠80只,随机均分为A、B、C、D组,以Impactor Model-Ⅱ打击器制备脊髓胸10(T10)损伤模型。A组为单纯损伤组,B组损伤后3d移植hUCMSCs,C组损伤后1w移植hUCM-SCs,D组损伤后3w移植hUCMSCs,分别于造模后24h、移植前1d及移植后8w(A组与D组损伤后相应时间点相同)每周对各组大鼠后肢功能进行行为学评分(BBB评分);移植后8周免疫组化染色观察移植细胞的存活、分化和血管再生情况,10周时用生物素葡聚糖胺(biotin dextran amine,BDA)示踪皮质脊髓束(corticospinaltract,CST)观察轴突的再生情况。结果:SCI后24h各组大鼠BBB评分无显著性差异(P0.05),D组在移植前和移植后1周与A组相应时间点的BBB评分无显著性差异(P0.05),移植后2w起各时间点评分均明显高于A组相应时间点,差异有统计学意义(P0.05);A组在SCI后9w时到达平台期,与实验结束时相比差异无统计学意义(P0.05)。移植各组移植后BBB评分逐渐上升,后一个时间点与同组前一时间点比较差异有显著性意义(P0.05),移植后6w开始C组明显高于B、D组(P0.05)。免疫组化染色A组脊髓中未见Brdu阳性细胞,损伤区血管形态欠完整,未见新生血管;损伤周围星形胶质细胞大量增生,细胞肥大、突起增多、排列不规则,形成胶质瘢痕;未见BDA标记的皮质脊髓束通过损伤区。B、C、D组脊髓中均有Brdu阳性标记的细胞存活,C、D组中细胞数量明显多于B组,但三组中均未见到移植细胞向神经元或神经胶质样细胞分化;B、D组损伤周围的星形胶质细胞较A组减少,但排列仍欠规则;C组损伤周围星形胶质细胞形态趋于正常,排列规则,且有大量星形胶质细胞通过损伤区,血管再生数目及通过损伤区的皮质脊髓束神经纤维也明显多于B、D组。结论:大鼠SCI后局部移植的hUCMSCs能长期存活,抑制胶质瘢痕形成,改善后肢运动功能;SCI后1w移植的效果优于SCI后3d和3w时移植的效果。     

密盖息对骨质疏松大鼠治疗作用的实验研究

目的观察密盖息单独和联合中药龟鹿补肾液治疗对骨质疏松大鼠骨密度及骨代谢的影响,探讨密盖息对骨质疏松的治疗作用以及与血钙、磷、维生素D代谢的关系,以及与生长因子的关系。方法用摘除大鼠双侧卵巢的方式制备骨质疏松模型,用密盖息治疗4周后,换用中药龟鹿补肾液治疗4周,应用HOLOGIC第4代双能X线4500W骨密度仪测大鼠全身、腰椎、股骨上段骨密度值(BMD),用ELISA法测定血清IGF-1水平和血清25OHVitD浓度以及血淋巴细胞维生素D受体(VDR)含量。结果密盖息治疗4周,治疗Ⅰ组(OVX+密盖息组)和治疗Ⅱ组(OVX+密盖息+中药组)较模型对照组(OVX组)腰椎、股骨上段骨密度增高,组间之比有显著性差异(P<0.01,或0.01相似文献   

金雀异黄酮对去势大鼠骨质疏松的影响

目的观察金雀异黄酮对实验性骨质疏松大鼠骨密度(bonemineral density,BMD)和血清骨代谢生化指标的影响。方法切除SD雌性大鼠双侧卵巢建立骨质疏松动物模型,给予金雀异黄酮皮下注射,治疗8w后监测大鼠腰椎及股骨的骨密度,检测血清骨碱性磷酸酶(ALP)、血清钙(Ca)、血清磷(P)等骨代谢生化指标。结果去势后的大鼠腰椎及股骨的骨密度降低;采用金雀异黄酮治疗后骨密度明显升高,与阴性对照组的比较差异具有统计学意义(P<0.05),与假手术组比较无显著性差异。假手术组和金雀异黄酮组血清ALP活性低于阴性对照组。结论金雀异黄酮可调节骨代谢,显著提高骨质疏松大鼠的骨密度。     

脊髓损伤对大鼠骨转换及骨密度的影响

目的：探讨脊髓损伤（spinal cord injury,SCI）对大鼠骨转换及骨密度的影响。方法：60只3月龄SD大鼠均分为SCI组与对照组。SCI组于T10处完全横断脊髓;对照组仅行椎板切除术。术后1、3、6周时处死动物测血清钙（Ca）、磷（P）、碱性磷酸酶（ALP）、尿钙、磷、肌酊（Cr）以及股骨、股骨骨密度（bone mineral density,BMD）。:桂花 血钙、尿钙、尿钙／肌酝在伤后不同时间段均升高;血ALP在伤后1周时显著下降,3、6周时恢复正常。胫骨近端,股骨远、近端的BMD在6周时较对照组下降且差异显著。结论：SCI大鼠可见明显的骨转换增高以及破骨活性增强,其生化、骨密度的改变与人体有较好的相关性。SD大鼠可用于评价SCI后骨代谢改变,具可以作为SCI后骨质疏松的模型。     

电针对去卵巢大鼠骨密度及生物力学的影响

目的研究电针对去卵巢骨质疏松模型大鼠骨密度(BMD)、生物力学的影响。方法选取60只雌性SD大鼠,随机分为空白组、模型组、假手术组、药物组、电针组,每组12只。空白组不做任何处理;假手术组只暴露双侧卵巢但不切除;其余3组行双侧切卵巢法建立骨质疏松模型大鼠。造模13周后开始治疗,电针组取"肾俞"、"脾俞"、"三阴交"、"足三里",每次电针刺激15 min(断续波,2 Hz,1.0m A)并灌服与药物组相同体积的蒸馏水,每日1次,连续治疗5 d,间隔2 d,20 d为1个疗程,共3个疗程;药物组大鼠以戊酸雌二醇水溶液(0.02 mg/mL,1 mL/100 g体质量)灌胃,疗程同电针组;空白组不予处理,常规饲养;模型组、假手术组大鼠灌服与药物组相同体积的蒸馏水,疗程同电针组。治疗结束后麻醉处死大鼠,分离股骨和第4腰椎,双能X射线骨密度检测仪检测股骨和腰椎BMD;三点弯曲法和压缩法检测股骨和腰椎生物力学。结果与假手术组相比,模型组大鼠股骨和第四腰椎离体BMD、股骨和第四腰椎的最大载荷及弹性模量均显著性降低(P0.01);与模型组相比,电针组、药物组大鼠股骨和第四腰椎离体BMD、股骨和第四腰椎的最大载荷及弹性模量均显著性升高(P0.01);电针组、药物组大鼠股骨和第四腰椎离体BMD、股骨和第四腰椎的最大载荷及弹性模量均显著性低于假手术组(P0.01)。结论针刺能有效增加大鼠骨密度,阻止骨量降低,减缓骨力学的退变,增强骨强度,对绝经后骨质疏松症具有一定的治疗作用。     

骨碎补总黄酮对去卵巢大鼠骨组织Cbfa1 mRNA表达的影响

目的观察成骨细胞转录因子Cbfa1在去卵巢骨质疏松(Osteoporosis,OP)大鼠的表达,探讨骨碎补总黄酮调节骨代谢的作用机制。方法 36只Wistar雌性大鼠随机均分为假手术组、模型组和药物组,术后2、4、6、8周检测血液雌二醇(E2)水平,进行股骨和腰椎骨密度测量、组织学观察和Cbfa1mRNA表达。结果在每个时间点3组大鼠雌二醇水平均存在显著性差异(P<0.05),假手术组>模型组>对照组;比较股骨及腰椎骨密度,假手术组>模型组(P<0.05)在每一个检测的时间点,药物组在术后2周和术后8周低于假手术组(P<0.05),在术后6周和术后8周高于模型组(P<0.05),其余时间点无显著性差异。组织学检查示模型组骨组织内骨小梁减少,稀疏,间隔增大,药物组该表现较少,而假手术组骨组织内未见该现象。模型组与药物组Cbfa1 mRNA表达在术后4周均低于假手术组(P<0.05),在术后6周和术后8周,假手术组>药物组>模型组(P<0.05)。结论实验结果表明雌激素减少能导致大鼠Cbfa1表达降低,而骨碎补总黄酮能上调Cbfa1表达水平,促使骨髓间质干细胞向成骨细胞分化发挥其防治OP作用。     

音猬因子修饰的骨髓间充质干细胞治疗大鼠骨质疏松的实验研究

目的探讨转染音猬因子的骨髓间充值干细胞移植对卵巢切除骨质疏松大鼠的治疗作用。 方法选取40只雌性SD大鼠，分为假手术组、模型组、骨髓间充值干细胞（BMSCs）移植组和音猬因子修饰的骨髓间充值干细胞（Shh-BMSCs）移植组，每组10只动物。假手术组手术中仅摘除卵巢附近的脂肪组织，但不摘除卵巢。其余的3组均进行双侧的卵巢切除手术，建立骨质疏松模型，在骨质疏松造模后的2个月，BMSCs移植组和Shh-BMSCs移植组分别通过大鼠的尾静脉注射BMSCs与Shh-BMSCs（第四代，约2*10⁶个干细胞），模型组动物注射等量PBS液体。在干细胞移植2 w后，检测尾静脉中血清钙、磷、碱性磷酸酶水平，同时采用双能X射线骨密度仪测量大鼠的股骨、腰椎及全身骨密度，micro-CT扫描进行胫骨的形态计量学指标分析。 结果（1）与假手术组相比，模型组的大鼠骨小梁厚度、骨小梁数量和骨体积分数均明显减少，大鼠的骨密度、血清钙与碱性磷酸酶明显降低，而大鼠的血清磷水平明显升高，差异有统计学意义（P<0.05），以上表明骨质疏松大鼠模型成功制备。（2）与模型组相比，BMSCs组的大鼠血清钙和和碱性磷酸酶明显升高，而血清磷水平明显降低，差异有统计学意义（P<0.05）。与模型组、BMSCs组相比，Shh-BMSCs组大鼠血清钙、碱性磷酸酶水平明显升高，而血清磷水平明显降低，差异有统计学意义（P<0.05）。（3）同模型组的骨密度相比，BMSCs组大鼠的骨密度明显升高，差异有统计学意义（P<0.05）；与模型组、BMSCs组相比，Shh-BMSCs组大鼠的骨密度明显升高，差异有统计学意义（P<0.05）。（4）同模型组相比，BMSCs组的大鼠骨小梁厚度、骨小梁数量和骨体积分数均明显增加，差异有统计学意义（P<0.05），与模型组和BMSCs组相比，Shh-BMSCs组的大鼠骨小梁厚度、骨小梁数量和骨体积分数均明显增加，差异有统计学意义（P<0.05）。 结论Shh-BMSCs细胞移植后能够治疗大鼠卵巢切除后的骨质疏松。     

脊髓损伤早期及制动后大鼠股骨远端骨量丢失的实验研究

目的:建立大鼠脊髓损伤及后肢废用模型,观察建模早期两种模型大鼠股骨远端骨密度及远端破骨细胞数量的改变。方法:24只SD大鼠随机分为3组(每组8只),对照组(A组),行T10椎板切除,不损伤硬膜及脊髓;脊髓损伤(SCI)组(B组):切除T10椎板后用Allen′s法(60g.cm势能)造成脊髓损伤;制动组(C组):采用大鼠双侧腿-尾缝合,造成双下肢制动。分别于造模后第1、7天采用BBB(Basso,Beattie and Bresnahan)评分评估脊髓损伤大鼠后肢运动功能,确定模型建立成功。建模第10天后处死,取一侧尺、桡骨及股骨远端,行骨密度检测。取另一侧股骨远端行TRAP染色,观察大鼠股骨远端破骨细胞数量的变化。结果:A、B、C 3组股骨远端骨密度值分别为0.114、0.096、0.108g/cm2,B组骨密度值最低,其次为C组,A组最高(P<0.05)。尺、桡骨骨密度3组比较无差异(P>0.05)。与A组比较,B组大鼠股骨干骺端破骨细胞数量增加显著,其次为C组,A组最少(P<0.01)。结论:脊髓损伤后,股骨远端破骨细胞数量大量增多,活性增强,骨吸收加强,引起脊髓损伤平面以下肢体骨量的迅速丢失,比废用性骨量丢失的程度更重。     

人脐带间充质干细胞与自体激活雪旺细胞联合移植修复脊髓损伤的实验研究

目的 通过观察人脐带间充质干细胞(hUCMSCs)和大鼠自体激活雪旺细胞(AASCs)联合移植修复脊髓损伤的疗效,探讨AASCs对hUCMSCs体内存活、分化的影响.方法 分离、培养hUCMSCs和大鼠AASCs.通过IMPACTOR MODEL-Ⅱ型打击仪将80只Wistar成年雌性大鼠均制作成T10损伤模型,随机分为四组(n=20):DMEM移植对照组、hUCMSCs移植组、AASCs移植组、hUCMSCs与AASCs联合移植组.比较符组动物恢复情况,进行行为学评分(BBB评分),NF-200和GFAP染色观察细胞存活、分化情况,生物素葡聚糖胺示踪观察皮质脊髓束再生情况.结果 4周后各组间BBB评分差异有统计学意义(P<0.05),6周后联合移植组明显高于其他三组,差异有统计学意义(P<0.05).免疫组化染色示联合移植组的hUCMSCs存活数量,NF-200、GFAP阳性荧光面积均明显高于hUCMSCs移植组,差异有统计学意义(P<0.05),BDA顺行爪踪可见联合移植组于损伤区染色较多,部分纤维延续至损伤远端.结论 AASCs可支持移植的hUCMSCs在损伤部位存活并向神经方向分化,hUCMSCs与AASCs联合移植较二者单独移植能更有效地促进脊髓损伤后运动功能的恢复和轴突再生.     

外伤性脊髓损伤患者的骨质疏松

为探讨脊髓损伤后骨质疏松的发生规律和特点,对83名外伤性脊髓损伤患者腰椎及髋部骨质疏松程度作放射学观察,井与正常对照者进行比较。结果:脊髓损伤患者的腰椎及髋部骨密度均较正常对照组有明显减低(P<0.01),但完全性瘫痪患者与不完全性瘫痪患者无显著统计学差异(P>0.05),骨密度参数与病程长短亦无显著统计学差异(P>0.05)。认为:脊髓损伤性骨质疏松与传统意义上的废用性骨质疏松有所不同,其发病机理及影响因素较为复杂,尚有待进一步研究。     

A study of spinal cord ischemia during aortic cross-clamp--evoked spinal cord potential and histological analysis of the spinal cord

The relationship between the evoked spinal cord potential (ESP) and the histological findings of the spinal cord after thoracic aortic cross-clamp was studied. Thoracic aorta was cross-clamped in 23 dogs and ESP was monitored before, during, and after cross-clamping. Incidence of paraplegia and histological findings were studied after the dogs recovered from the procedure. Aortic cross-clamp was maintained for 60 minutes in 20 dogs (Group A). And cross-clamp was released 10 minutes after the amplitude of ESP became lower than 20% of control in 3 dogs. (Group B). In group A, three types of ESP changes were detected; ESP became lower or lost during cross-clamping in type 1 response, ESP remained unchanged in type 2 response, and ESP returned after transient loss during cross-clamping in type 3 response. Four of five dogs with type 1, none of nine with type 2, two of five with type 3 response showed paraplegia. One of the dogs with type 2 response showed paraparesis. ESP could not detected in one dog, in which traumatic spinal cord injury during laminectomy caused paraplegia. In Group B, all dogs showed type 1 response and paraplegia. Characteristic histological finding of the spinal cords of the dogs with paraplegia was the ischemic necrosis mainly in the gray matter. Necrotic foci were limited in the posterior horn in mild, in the anterior and posterior horn in moderate changes. And neurons were lost in entire gray matter in severe histological changes. In the spinal cords of the dogs with spastic paraplegia, severe histological changes were limited in the lower lumbar region.(ABSTRACT TRUNCATED AT 250 WORDS)     

Progressive cystic degeneration of the spinal cord following spinal cord injury

From a group of 520 spinal cord injury patients treated at Rancho Los Amigos Hospital, two cases of progressive myelopathy secondary to cystic degeneration of the spinal cord have been identified. The cyst may dissect proximately to produce progressive neurologic deficit. Surgical treatment with shunting can allow stabilization and improvement with return of newly lost function.     

Liposarcoma of spermatic cord

Liposarcoma of the spermatic cord is a rare tumor. Most are of a low-grade malignancy, and treatment is wide local excision with radical orchiectomy. At present there is no clear indication for chemotherapy. Because recurrence, especially local recurrence, is a possibility, periodic followup is necessary. Local recurrences are treated with repeat wide local excision and radiotherapy.     

Rhabdomyosarcoma of spermatic cord

Rhabdomyosarcoma is the most common malignant spermatic cord tumor. Thus far 162 cases have been reported. Herein, we present 4 additional cases, with a five-year survival rate of 75 per cent using a multimodal treatment program.     

Liposarcoma of spermatic cord

Four patients with myxoid liposarcoma of the spermatic cord are presented, three of whom have survived longer than five years. Wide local excision is usually the treatment of choice with radiotherapy reserved for control of either extensive local disease or following incomplete removal of the lesion.     

Torsion of spermatic cord

A series of 198 cases of acute unilateral, painful scrotal swelling considered to be torsion is presented and an aggressive surgical management of this entity described. The results achieved, a high incidence of testicular salvage, justify this aggressive approach.     

Leiomyosarcoma of spermatic cord

A case of leiomyosarcoma of the spermatic cord is presented, bringing the total number of reported cases to 39. A review of diagnostic and therapeutic approaches along with our experiences stress the need for a uniform treatment schedule.     

Rhabdomyosarcoma of spermatic cord

A case of embryonal rhabdomyosarcoma of the spermatic cord is presented and literature reviewed. The spread of this disease to the retroperitoneal lymph nodes and the improved results with retroperitoneal lymphadenectomy and radical orchiectomy are stressed.