神经干细胞原代培养及GABA能神经元的诱导分化

目的探讨神经干细胞(NSC)的原代培养及γ-氨基丁酸(GABA)能神经元的诱导分化、鉴定。方法取出生1~3dSD大鼠全脑,分离、培养NSC并诱导分化了GABA能神经元,用免疫荧光染色进行鉴定。结果培养24h后,由单细胞发展成2~4细胞神经球,随时间延长球体增大,神经球均有Nestin阳性细胞;用含10%血清分化液分化1d后,神经球开始贴壁,伸出细长突起,部分可和周边神经球伸出的突起连接。神经球周边见大量散在贴壁的双极或多极细胞,免疫荧光染色可见NSE、GABA阳性细胞。加了分化液的实验组GABA阳性细胞分化率为(30·25±2·12)%,而对照组分化率为(1·14±1·39)%。结论成功从新生大鼠脑组织中分离、培养了NSC,并能较大比率分化GABA能神经元。     

GDNF和IL-1β体外诱导中脑神经干细胞分化的实验研究

目的探索胶质源性神经营养因子（GDNF）和白细胞介素1β（IL-1β）在体外诱导小鼠胚胎中脑神经干细胞（M-NSCs）分化成多巴胺能神经元的培养方法,为M-NSCs移植治疗帕金森病（PD）提供实验依据。方法在有血清条件下体外培养鼠胚M-NSCs,予以GDNF和IL-1β作诱导分化。TH免疫细胞化学鉴定,流式细胞术检测TH阳性神经元的百分率。结果实验组促进M-NSCs分化为TH阳性神经元的比例：A组为13.53%,B组为9.66%,C组为16.22%,D组（对照组）仅3.46%,有显著性差异（P〈0.01）。结论 GDNF和IL-1β可明显促进M-NSCs分化成多巴胺能神经元。     

PD大鼠纹状体提取液诱导中脑神经干细胞分化为多巴胺能神经元的研究

目的研究PD大鼠纹状体提取液诱对中脑神经干细胞的诱导分化作用.方法①体外分离、培养大鼠胚胎的中脑神经干细胞,并诱导其分化,巢素(Nestin)、神经丝-200(NF-200)、胶质纤维酸性蛋白(GFAP)免疫组化染色鉴定.②加入纹状体提取液后,分化细胞用抗酪氨酸羟化酶(TH)免疫组化鉴定,并用流式细胞技术检测分化比例.结果①从大鼠胚胎的腹侧中脑部位分离获得了大量未分化、呈巢状悬浮生长的细胞团,且能分化为神经元和神经胶质细胞.②经纹状体提取液诱导后,能够分化出多巴胺能神经元,分化比例达20%左右.结论纹状体提取液能诱导中脑神经干细胞向多巴胺能神经元的分化.     

人孤雌胚胎干细胞向多巴胺能神经元的体外分化

目的 研究人孤雌胚胎干细胞向神经上皮祖细胞和神经元诱导分化的能力.方法 人孤雌胚胎干细胞在人饲养层细胞上诱导后机械消化传代,在多巴胺能神经元诱导培养基中悬浮后贴壁分化,Pax6、Nestin和PSA-NCAM抗原染色检测神经上皮祖细胞的形成,Tuj1和TH抗原粢色检测多巴胺能神经元的形成.结果 人孤雌胚胎干细胞诱导可形成的神经上皮祖细胞中,具有典型的rosette结构,其Pax6和Nestin阳性细胞比例分别为(84.7±5.3)%和(92.0±4.7)%.该神经上皮祖细胞可形成PSA-NCAM染色阳性的神经元前体和Tuj1染色阳性的神经元,Tuj1阳性细胞中(33.3±4.3)%为酪氨酸羟化酶染色阳性细胞.结论 人孤雌胚胎干细胞体外诱导分化可形成高纯度的神经上皮祖细胞,并可进一步分化为多巴胺能神经元.     

小鼠胚胎干细胞体外诱导分化成GABA能神经元

目的探讨小鼠胚胎干细胞在体外培养向GABA能神经元定向诱导分化的可能性。方法将小鼠胚胎干细胞以"无血清"方法培养,用DMEM/F12、N2、B27及NT4作为诱导分化剂定向诱导分化,分化好的细胞利用免疫荧光技术、流式细胞技术和RT-PCR鉴定。结果在胚胎干细胞诱导分化成神经元后期,免疫荧光显示有GABA能神经元存在;RT-PCR结果证实有GABA能神经元正确分化的重要调控基因Viaat、Gad1和Gad2基因表达;流式细胞仪计数结果显示GABA阳性细胞约占总细胞数的(11.49±6.86)%。结论小鼠胚胎干细胞经体外培养可以定向诱导分化成GABA能神经元,可作为神经移植的新来源。     

清脑灵诱导大鼠骨髓间充质干细胞分化成神经元样细胞

目的观察清脑灵煎剂是否能诱导大鼠骨髓间充质干细胞(MSCs)分化成神经元样细胞。方法通过贴壁法分离获得大鼠骨使髓间充质干细胞,体外扩增培养,用流式细胞仪检测细胞表面抗原的表达。用浓度为1mg/mL的清脑灵煎剂诱导MSCs,分别于5h、12h、1d、3d、7d在倒置显微镜下观察细胞形态变化,用免疫细胞化学方法观察被诱导细胞巢蛋白(nestin)、神经元特异性烯醇化酶(NSE)、神经胶质纤维酸性蛋白(GFAP)的表达。结果流式细胞仪检示MSCs不表达CD11b、CD45,诱导后细胞的免疫细胞化学示nestin阳性细胞、NSE阳性细胞和GFAP阴性细胞。结论清脑灵能诱导MSCs分化神经元样细胞。     

人神经干细胞向多巴胺神经元的定向分化

目的 :将体外扩增的神经干细胞在体外诱导向多巴胺能神经元分化 ,为帕金森氏病的细胞移植治疗奠定基础。方法 :利用无血清培养和单细胞克隆技术从 4月龄胚胎中脑组织获得神经干细胞 ,用 10 %胎牛血清 (FBS)、IL -1α以及IL -1α +IL -11+LIF+GDNF的组合诱导神经干细胞向多巴胺能神经元分化 ,用酪氨酸羟化酶 (TH)免疫检测分化为多巴胺神经元的情况。结果 :10 %FBS只能使极少数神经干细胞分化为TH阳性的多巴胺神经元 ;IL -1α可以诱导神经干细胞较多地分化为TH阳性多巴胺神经元 ,但细胞形态上不成熟 ;联合应用IL -1α、IL -11、LIF、GDNF后 ,分化的多巴胺神经元数量上和单用IL -1α相似 ,但细胞形态上比较成熟。结论 :联合应用IL -1α、IL -11、LIF、GDNF等细胞因子诱导人神经干细胞分化得到的TH阳性多巴胺神经元不但数量较多 ,其细胞形态也比较成熟     

全反式维甲酸及银杏提取物对神经干细胞分化的影响

目的探讨全反式维甲酸(ATRA)、银杏提取物(Egb)对体外培养的神经干细胞(NSCs)分化为胆碱能神经元和多巴胺能神经元的作用。方法取第3代NSCs诱导分化,试验分为4组:ATRA组(10-8mol/L)、Egb组(2mg/L)、ATRA(10-8mol/L)+Egb组(2mg/L)和空白对照组;免疫细胞化学染色检测神经微管相关蛋白(MAP2)、乙胆碱酯酶(AchE)和酪氨酸羟化酶(TH)的表达,并计算阳性染色细胞的百分率。结果ATRA组诱导神经干细胞分化为AchE阳性和TH阳性神经元的百分率分别为26·32%±3·62%和12·02%±2·78%;Egb组分别为42·93%±3·08%和18·13%±1·74%;ATRA+Egb组分别为43·32%±5·60%和13·03%±2·54%;空白对照组分别为27·68%±2·51%和5·74%±1·81%。结论ATRA和Egb均可促进NSCs向TH阳性神经元分化,Egb还可促进NSCs向AchE阳性神经元分化,ATRA和Egb联用无协同效应。     

全反式维甲酸诱导中脑神经干细胞向多巴胺能神经元分化的作用

目的：观察全反式维甲酸（all-trans retinoic acid,ATRA）体外对大鼠中脑神经干细胞（neural stem cells,NSCs）的诱导分化作用及其可能的分子机制。方法：分离培养孕14～15 d大鼠中脑NSCs,使用1.0μmol/L ATRA诱导1、3、5和7 d,以免疫组织化学染色观察ATRA对NSCs分化为多巴胺能神经元的影响;RT-PCR分析维甲酸（retiadic acid,RA）各受体表达情况。结果：ATRA诱导后多巴胺能神经元特异性酪氨酸羟化酶（TH）染色呈阳性,细胞有多个细长突起,与对照组比较,诱导7 d后多巴胺神经元特异性TH染色阳性细胞率均明显升高（P〈0.01）;各诱导组RA受体βmRNA的表达量均明显升高（P〈0.01）,而且存在时间依赖性。结论：ATRA能显著提高NSCs分化为多巴胺能神经元的比例,同时发现细胞维甲酸A受体（RAR）βmRNA表达增加,RARβ激活可能促进NSCs向多巴胺神经元方向的分化。     

全反式维甲酸促人脐血多能干细胞向多巴胺能神经元分化

目的探讨在全反式维甲酸(all-trans retinoic acid,ATRA)作用下,人脐血多能干细胞(cord blood-stem cells,CB-SCs)向多巴胺能神经元的分化潜能。方法分离、培养CB-SCs后,分为诱导组、神经培养基对照组和无血清培养基对照组。诱导培养12 d后,观察干细胞的形态学变化,免疫化学荧光法检测多巴胺神经元标志物的表达。ELISA法检测上清液中多巴胺递质的含量。结果 ATRA处理后,大部分CB-SCs出现神经样分化,神经培养基对照组仅少量CB-SCs出现神经样分化,无血清培养基对照组CB-SCs继续扩增而无明显形态学变化;诱导组(48±11)%的细胞酪氨酸羟化酶(TH)表达阳性;神经培养基对照组仅少量TH阳性细胞;ELISA检测结果表明,诱导组多巴胺较神经培养基对照组明显升高(P<0.001)。结论 ATRA可以较为高效地促CB-SCs分化为多巴胺能神经元。     

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.