嗅鞘细胞移植促进急性脊髓损伤的修复作用

目的研究嗅鞘细胞(OEG)移植后在体内存活情况以及对大鼠脊髓损伤长期的修复作用.方法用新生Wistar大鼠嗅球做OEG培养,大量增殖并标记.80只大鼠,随机分为4组,每组20只.A、B、C组以250 mm·g打击T13脊髓致运动诱发电位和体感诱发电位完全消失,A组在打击区中央用尖刀横行切断脊髓;D组仅做与A、B组相同节段的椎板减压,不挫伤脊髓.A、B组在脊髓远近断端距离挫伤区边缘1 mm的脊髓中线上,深度分别为1.75、1.5、1.0、0.5 mm处,各注入200 000个OEG;C组用同样的方法注射等量的DMEM培养液;D组不做处理.术后观察脊髓功能恢复情况(BBB运动功能评分法)和体重变化.术后24周取脊髓标本,做HE染色、嗜银染色、抗神经原纤维(NF)免疫组化染色、荧光显微镜下观察Hoechst标记的OEG在体内存活情况.结果手术后4周,A、B、C组开始有运动功能恢复,术后4～24周BBB运动功能评分A、B组均高于C组(P＜0.01),16周以后BBB评分变化较小.HE染色A、B两组脊髓损伤区结构紊乱,纤维走行方向扭曲不一致,细胞数目较多,C组大鼠脊髓损伤段常有明显空洞存在,纤维含量少且扭曲.嗜银染色和免疫组化抗NF染色显示无论在损伤区还是损伤头、尾端,A、B组神经纤维数量多于C组,但都少于相同节段的D组(P＜0.01).荧光显微镜下观察到Hoechst标记的OEG大量存在于脊髓损伤段周围.术后各组大鼠均出现体重减轻.术后2周,D组大鼠体重恢复至术前重量,此后逐渐增加.术后4周,A、B、C组大鼠体重降至最低,与D组(Sham组)比较差异有显著性(P＜0.01);此后,A、B组大鼠体重逐渐增加,而C组大鼠体重增加不明显;A、B组大鼠体重与C组比较差异有显著性(P＜0.05).结论OEG移植具有一定促进脊髓损伤神经纤维再生和功能恢复的作用.     

他克莫司联合骨髓间充质干细胞移植对大鼠急性脊髓损伤后的再生修复实验研究

目的观察用他克莫司(tacrolimus,FK506)联合骨髓间充质干细胞(bone mesenchymal stem cell,BMSCs)移植对脊髓损伤后再生的影响,进一步探讨两者对急性脊髓损伤(spinal cord injury,SCI)的作用和机制。方法钳夹法制作SD(Spraque-Dawley)大鼠急性脊髓损伤动物模型30只,随机分成三组。A组为单纯损伤组,B组为单纯骨髓干细胞移植组,C组为骨髓干细胞联合FK506组。A、B、C三组一周后再次手术,暴露出原脊髓损伤处,A组将生理盐水10μl用微量注射器注入于脊髓损伤区域远近端;B、C两组将体外培养的骨髓间充质干细胞10μl用微量注射器分别注入于脊髓损伤远近端;C组同时另行腹腔再注射0.25%FK506一次,用量1mg/kg。期间加强动物的护理,于术前、术后1w、2w、4w、6w、8w采用BBB评分、斜板实验及后肢行为学评分观察神经恢复情况。8周后取材,行免疫组化检查及病理学检查。结果各组BBB评分B、C两组大鼠后肢运动功能恢复较明显,B、C组注射细胞后2周开始出现后肢活动,逐步恢复排尿,伤后4周变化最为明显,可见明显后肢活动和尾巴的摆动,伤后8周,已可见协调运动,C组的恢复情况较B组为快,BBB最高评分可达14分,A组亦有所恢复,但程度较轻。B、C两组评分高于A组,B、C两组统计学上有差异(P<0.05)。C组与A组在6周以后的评分有显著性差异(P<0.01)。斜板实验和后肢行为学评分与BBB评分类似。免疫组化显示:A组切片见伤处为瘢痕连接,结构紊乱,邻近有明显空洞形成,未见神经轴索通过。B、C组亦见瘢痕形成,但可见少量神经轴索样结构。A组伤处邻近神经元胞体NF200阳性细胞较少,邻近脊髓组织GFAP阳性细胞亦较少。B组伤处可见GFAP及NF阳性纤维,但排列较紊乱,无正常结构。C组较B组邻近脊髓神经元胞体NF200阳性细胞较多,GFAP阳性细胞亦较多。C组大鼠脊髓损伤边缘Nogo阳性神经元较A、B两组明显减少。结论脊髓损伤的大鼠移植的大鼠骨髓间充质干细胞在脊髓损伤部位能长期存活,并能向邻近脊髓内迁徙,骨髓间充质干细胞移植对于后肢功能的恢复有促进作用;联合应用FK506有协同效果;脊髓损伤后功能恢复需要一个多种神经因子共同参与的复杂的微环境。     

微囊化异种嗅球组织移植对脊髓损伤大鼠细胞凋亡及功能恢复的影响

目的 观察微囊化异种嗅球组织细胞移植对大鼠脊髓全横断损伤后细胞凋亡及功能恢复的影响,探讨其对脊髓继发性损伤的保护作用.方法 横断SD大鼠T10节段脊髓,建立脊髓全横断损伤动物模型.在无菌条件下取家兔双侧嗅球,制备成细胞悬液及微囊化的细胞悬液.实验大鼠随机分为4组:正常组(A组)仅打开椎管,暴露脊髓;损伤组(B组)仅用明胶海绵填塞脊髓损伤腔隙;细胞悬液组(C组)用明胶海绵吸附的兔嗅球组织细胞悬液植入;微囊组(D组)植入明胶海绵吸附的微囊化兔嗅球组织细胞悬液.实验大鼠分别于术后12 h、1 d、3 d、7 d、21 d进行神经功能评分(BBB法)评分评价大鼠运动功能的恢复情况,并行TUNEL染色,观察凋亡细胞的数量及分布变化.结果 A组少见凋亡细胞;B组凋亡细胞最多,并于损伤后1d、7d形成两个高峰;C组凋亡细胞数量较B组有所减少,差别具有显著性(P＜0.05).C组BBB评分较B组明显提高,有显著性差别(P＜0.05);D组凋亡细胞数量较B组明显减少,差别非常显著(P＜0.01),较C组有所减少,差别具有显著性(P＜0.05).D组BBB评分较B组明显提高,差别非常显著(P＜0.01),较C组有所增加,差别具有显著性(P＜0.05).结论 异种嗅球组织细胞悬液移植能够抑制细胞凋亡的发生,能够促进脊髓损伤大鼠的功能恢复,用微囊化异种嗅球组织细胞移植效果更佳.     

cAMP衍生物特异性激活Epac2/Akt信号通路促进大鼠脊髓损伤的修复

目的 研究应用cAMP衍生物特异性激活Epac2/Akt信号通路在哺乳动物脊髓损伤中的病理和生理机制.方法 96只成年健康雌性SD大鼠,完全随机分为4组(n=24),A、B、C组均采用改良Allen重物打击法制备大鼠脊髓损伤模型.A组给予8-CPT-2’-O-Me-cAMP 0.5 mg/(kg·d)、B组给予等量10％(体积分数)DMSO、C组给予8-CPT-2’-O-Me-cAMP 0.5 mg/(kg·d)加LY294002 0.2 mg/(kg·d)各7d.D组为正常对照组.各组分别于1、2、3、4周采用BBB评分法评估大鼠后肢功能恢复情况,各时间点取材行HE染色、免疫荧光、实时荧光定量PCR检测脊髓细胞标志物(Nestin、NF200、GFAP)和通路因子(Epac2、Akt)的表达.结果 A组大鼠在术后1～4周BBB评分持续升高,均高于B、C组(P＜0.01),低于D组(P＜0.01).在1、2、3周,A组Epac2 mRNA表达与C组Epac2 mRNA表达差异无意义,但高于B组(P＜0.01)和D组(P＜0.01).A组Akt mRNA表达高于B、C、D组(P ＜0.01).A组Nestin、NF200 mRNA表达在各期均高于B、C、D组(P＜0.01).A、B、C组GFAP mRNA表达差异无统计学意义.HE染色可见A、B、C组造模后脊髓组织结构疏松,有大量空洞形成.免疫荧光染色并进行积分光密度值分析,其结果与实时荧光定量PCR检测结果相符.结论 在哺乳动物脊髓损伤后应用cAMP衍生物能够特异性激活Epac2/Akt通路,促使神经干细胞分化和神经细胞增殖,有助于脊髓修复.     

参芎葡萄糖注射液对脊髓损伤后运动功能恢复及GAP-43和NF200表达的影响

目的探讨参芎葡萄糖注射液对急性脊髓损伤后运动功能恢复的影响和可能机制。方法应用改良的Allen’s法复制大鼠脊髓损伤模型60只,随机分为对照组(30只)和治疗组(30只),在建模后1、7、14、21和28 d用BBB评分和斜板试验评定脊髓损伤后后肢运动功能恢复并获取损伤段脊髓标本,用免疫组织化学方法和免疫印迹法检测两组脊髓在损伤后不同时段生长相关蛋白(GAP-43)和神经丝蛋白(NF200)的表达,对BBB评分、GAP-43、NF200进行相关分析。结果 BBB评分和斜板试验结果提示,治疗组大鼠7、14、21、28 d时的神经功能恢复较对照组明显提高(P<0.05);免疫组织化学方法和免疫印迹法提示,与对照组相比治疗组在一定时间段能上调损伤脊髓区GAP-43和NF200的表达(P<0.05);在术后7 d和14 d,BBB评分与GAP-43、NF200的表达正相关,在术后7、14、21、28 d GAP-43与NF200的表达正相关。结论参芎葡萄糖注射液能促进损伤脊髓的功能恢复,其机制可能与参芎葡萄糖注射液上调损伤区脊髓GAP-43、NF200的表达有关。     

CDNF治疗大鼠脊髓损伤的实验研究

目的研究CDNF对于大鼠脊髓损伤的治疗效果。方法选择Wista雌性大鼠75只,将其中25只设置为假手术组(A组),25只设定为单纯脊椎损伤组(B组),25只设定为采用CDNF治疗脊髓损伤组(C组)。在大鼠的T9~10节段使用脊髓半横断法制作脊髓损伤的模型,其中在B组大鼠的脊髓损伤处使用生理盐水,在C组大鼠的脊髓损伤处使用CDNF。在手术后对3组大鼠的进行应用行为学评分,对大鼠的脊髓损伤的修复情况进行组织形态学观察,采用免疫组织化学方法观察各组大鼠的GFAP表达的变化。结果 C组大鼠的应用行为学评分明显高于B组,但是仍然低于A组。C组的GFAP的表达低于B组。C组大鼠的GFAP阳性细胞低于B组,差异有统计学意义(P<0.05)。结论 CDNF对于脊髓损伤后反应性星形胶质瘢痕性的形成有明显的抑制作用,进而能够促进突触的连接重建,加速神经功能的恢复。     

外源性血管内皮生长因子对损伤脊髓的作用

目的 研究外源性血管内皮生长因子(VEGF)局部应用对大鼠损伤脊髓神经组织的作用.方法采用改良Allen氏重量打击法大鼠急性脊髓损伤模型.脊髓打击性损伤后,将模型大鼠随机分组,并依组别在蛛网膜下腔分别给药.A组:生理盐水10μL,B组:Matrigel蛋白胶10 μL,C组:重组人VEGF165 0.2 μg+Matr...     

银杏提取物对鼠脊髓损伤后神经细胞的保护及功能恢复作用

目的:研究银杏提取物(GBE)对大鼠脊髓损伤后神经细胞的保护及功能恢复作用。方法:75只W istar雄性大鼠,随机分为假手术组(A),对照组(B)和治疗组(C),每组25只,A组只打开椎板,不损伤脊髓,B组脊髓损伤后腹腔注射等渗盐水(NS),C组脊髓损伤后1 h腹腔注射GBE 40 mg/kg,此后每天1次,持续治疗5 d。末次治疗后每组处死5只动物,检测脊髓组织SOD活性,MDA和NO含量,脊髓损伤后第1 h和末次治疗后第1天、第5天、第10天评定后肢运动功能(Salzm an评分)和检测MEP。结果:SOD活性C组高于B组(P<0.05),MDA和NO含量C组低于B组(P<0.05)。脊髓损伤后,后肢运动障碍,MEP波幅(P1-N1峰峰值)降低,潜伏期延长,末次治疗后各时间,治疗组MEP波幅、潜伏期,神经功能恢复均优于对照组(P<0.05)。结论:银杏提取物能减少脊髓损伤后氧自由基所致继发损伤,并促进神经功能恢复。     

O-2A祖细胞移植对大鼠脊髓损伤后运动功能恢复的作用

目的 研究O-2A祖细胞移植对大鼠脊髓损伤(SCI)后运动功能恢复的作用.方法 从新生1～2 d SD大鼠大脑皮质中分离纯化O-2A祖细胞.80只SD成年大鼠,随机分为4组:A组为正常组,与其他3组同期饲养;其他3组均行T9脊髓左半侧离断手术,B组损伤处植入吸附生理盐水的明胶海绵(损伤对照组);C组损伤处移植50%O-2A祖细胞和50%1型星形胶质细胞(T1A)混悬液(O-2A+T1A组);D组损伤处植入含有纯化O-2A祖细胞的细胞悬液(O-2A移植组).移植术后3、7、14、21、28 d分别以CBS及BBB方法 进行大鼠神经功能评定.结果 A组大鼠后肢CBS和BBB得分分别为100和21.O-2A祖细胞移植术后3、7、14、21、28 d,B、C、D组动物后肢神经功能均有不同程度的恢复,左后肢得分均显著低于右侧,差异均有高度统计学意义(均P<0.01),D组运动功能恢复最好.结论 O-2A祖细胞移植可促进大鼠脊髓损伤后神经功能的恢复.     

NgR基因沉默细胞-PLGA移植对脊髓损伤大鼠后肢功能的影响

目的观察NgR基因沉默细胞-PLGA共移植对脊髓损伤大鼠后肢功能修复的作用。方法健康成年雌性Wistar大鼠72只,建立脊髓T9半横断损伤模型。随机分为单纯支架组(A组)、骨髓间充质干细胞+施旺细胞+聚乳酸-乙醇酸(PLGA)组(B组)、NgR基因沉默细胞+PLGA组(C组),每组24只。于脊髓缺损处分别植入对应的复合物。造模后1、2、4、6、8周行运动功能检测,4周后行HE染色及免疫荧光观察PKH-26标记的骨髓间充质干细胞的存活和分布情况。结果斜板实验和改良BBB评分结果提示,下肢运动功能评价B组优于A组(P<0.01),C组优于B组(P<0.05);A、B、C组PKH-26阳性细胞数分别为0、(34.43±9.78)、(69.58±8.15)个/高倍视,3组比较差异有统计学意义(P<0.01)。结论 NgR基因沉默细胞-PLGA共移植能够明显促进脊髓半横断损伤大鼠的脊髓轴突再生,并改善肢体的运动功能。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.