大鼠不同类型脑损伤后NO表达变化的比较研究

目的 研究不同类型脑损伤后ＮＯ的表达变化。方法 建立大鼠３种脑２模型：脑皮质针刺伤、短暂性余脑缺血／再灌注与模拟低压性脑缺氧,运用ＮＡＤＰＨ－ｄ组化（ＮＤＰ）、原位分子杂交、图像分析等对３种脑损伤后ＮＯ的表达及其细胞来源进行对比观察。结果（Ｆ１）ＮＤＰ阳性细胞数在脑皮质针刺伤后第５－７ｄ达高峰,且主要分布于海马、室周区等处;全脑缺血再灌注后１－３ｄ即达高峰,以颞叶皮质、海马与室周区明显;高原缺氧     

大鼠创伤性脑损伤后损伤区周围脑组织脑脂结合蛋白的表达变化

目的观察大鼠创伤性脑损伤（TBI）后不同时间点脑损伤区周围脑组织脑脂结合蛋白（BLBP）的表达变化。方法采用大鼠颅脑液压损伤模型,在伤后1、3、7、14 d用Western blot检测损伤区周围脑组织BLBP的表达,用免疫荧光双标法检测BLBP和波形蛋白。结果 TBI后1 d,BLBP含量显著下降,后逐渐上升,至伤后7 d达到高峰,伤后14 d又急剧下降（均P〈0.01）。BLBP和波形蛋白双标阳性细胞主要分布在损伤区周围皮质及损伤侧胼胝体,损伤侧皮质阳性细胞大多为反应性星形胶质细胞形态;损伤侧胼胝体阳性细胞基本呈放射状胶质细胞（RGCs）形态。结论 TBI后损伤区周围脑组织RGCs样细胞可能与皮质神经再生和神经功能恢复有关。     

大鼠缺血性脑损伤早期一氧化氮含量的变化及其细胞来源的研究

目的 观察缺血性损伤后脑内一氧化氮(Nitric oxide，NO)含量的变化并确定NO的细胞来源。方法 采用短暂性全脑缺血再灌注模型，运用NO生化检测、NADPH-黄递酶组化、免疫荧光双标技术及图像分析等方法。结果 ①脑内NO的浓度于缺血再灌注后12h后即升高，1～3d达高峰并持续至第5天，此后逐步下降。②NDP阳性细胞数在缺血再灌注后1d即达高峰，以颞叶皮质、海马与室周区明显；再灌注14d后阳性细胞减少或消失。③脑缺血早期(1～3d)NDP阳性细胞与GFAP免疫荧光阳性细胞少数有共存，仅占10％～15％。结论 缺血性脑损伤早期NO的浓度增高，合成NO的细胞以神经元为主。     

小胶质细胞对脑缺血的影响

小胶质细胞是中枢神经系统的重要免疫细胞,在脑缺血后的炎症反应过程中发挥炎性损伤和神经保护双重作用。脑缺血发生后,小胶质细胞出现活化和增生,并被激活为M1型和M2型,不同的小胶质细胞表型对脑缺血的作用不同。M1型极化的小胶质细胞通过释放促炎性介质抑制中枢神经系统恢复,而M2型极化通过释放抗炎细胞因子、转化生长因子、脑源性神经生长因子等促进组织的修复和再生。本文将对脑缺血后小胶质细胞的活化及机制、迁移增生、极化及小胶质细胞对脑缺血的影响作一综述。     

补阳还五汤及其有效部位组方对沙土鼠脑缺血后反应性星形胶质细胞活化的影响

目的：比较研究补阳还五汤及其有效部位组方对脑缺血后反应性星形胶质细胞活化的影响。方法：采用沙土鼠双侧颈总动脉夹闭脑缺血模型,分别腹腔注射补阳还五汤及其有效部位组方,于不同时间取脑组织,以 免疫组化方法测定胶质纤维酸性蛋白（GFAP）的表达。结果：缺血15min后各组脑组织无GFAP表达。缺血15min再灌注24h后,脑组织GFAP表达达高峰,主要在海马区表达,补阳还五汤及其有效部位组方可使GFAP表达减弱,两者作用相近。缺血15min再灌注48h后,GFAP表达有所减弱,而补阳还五汤及其有效部位方组GFAP表达则较模型组有所增强,两者作用相近。结论：星形胶质细胞活化主要在脑缺血后再灌注期,在再灌注早期,补阳还五汤及其有效部位组方通过对抗缺血性脑损伤而改善星形胶质细胞功能的紊乱;在再灌注后期,则通过促进星形细胞的活化,利于损伤的修复。     

芹菜素对急性局灶性脑缺血再灌注损伤大鼠半暗区内小胶质细胞的影响

目的 探讨芹菜素对急性局灶性脑缺血再灌注损伤大鼠半暗区内小胶质细胞的影响及意义。方法 采用改良线栓法建立大鼠局灶性脑缺血再灌注模型。动物随机分为假手术组、模型组、芹菜素组。模型组和芹菜素组按再灌注时间不同又分为再灌注6、24、48、72 h及7 d各5个小组,共11组。各组观察至规定的时间进行神经行为学评分,其中随机取6只大鼠,进行异硫氰酸荧光素标记的同工凝集素B4（FITC-ILB4）染色,激光扫描共聚焦显微镜（LSCM）观察浸润的单核细胞及半暗区小胶质细胞形态及数量的变化,余下6只大鼠取脑组织,测定脑含水量。结果 脑缺血再灌注6 h,模型组大鼠脑缺血半暗区周围出现同工凝集素B4阳性（ILB4⁺）表达细胞即分枝状小胶质细胞（罗丹明6G标记阴性即Rhod 6G^?）和浸润的单核细胞（罗丹明6G标记阳性即Rhod 6G⁺）;48 h小胶质细胞明显增多,形态向阿米巴样转变;72 h达到高峰,主要为脑源性小胶质细胞增生,形态以阿米巴样为主;7 d小胶质细胞减少;芹菜素组相应时间点小胶质细胞较模型组明显减少(P＜0.05、0.01),形态亦呈现相似改变。脑缺血48、72 h,芹菜素组大鼠脑组织含水量明显低于模型组(P＜0.01)。模型组神经行为学评分与半暗区ILB4⁺细胞数呈负性相关（r＝?0.415,P＜0.05）,且芹菜素能有效降低模型组神经功能缺损程度(P＜0.05),明显减轻脑损伤程度。结论 缺血后的大脑脑实质内一部分小胶质细胞来源于循环中的单核细胞;半暗区小胶质细胞形态及数量的变化与脑损伤程度具有相关性;芹菜素对急性局灶性大鼠脑缺血再灌注损伤具有保护作用,且可能与下调小胶质细胞数量,抑制小胶质细胞过度活化密切相关。     

GFAP、S-100蛋白、波形蛋白在大鼠前脑缺血再灌注后的分布

目的　探讨波形蛋白 (Vimentin)、胶质纤维酸性蛋白 (Glialfibrillaryacidicprotein ,GFAP)和S -10 0蛋白在脑缺血再灌注后反应性星形胶质细胞的活化情况。方法　采用免疫组织化学方法 (SABC法 ) ,利用前脑缺血再灌注模型检测细胞活化情况。结果　缺血再灌注后第 7天 ,少量GFAP阳性细胞开始出现于小脑 ;缺血再灌注 2 0天时 ,GFAP阳性细胞数会略有增加。S -10 0蛋白免疫反应在第 7天已经明显 ;波形蛋白免疫反应在再灌注第 7天和第 2 0天时明显存在于小胶质细胞和寡突胶质细胞。结论　GFAP阳性细胞、小胶质细胞、寡突胶质细胞等参与了脑损伤后神经元的修复。     

大鼠坐骨神经损伤后早期脊髓背角小胶质细胞活化状态和活化类型的变化规律

目的 研究坐骨神经损伤后脊髓背角小胶质细胞活化状态和活化类型的变化规律。方法 大鼠随机分为对照组(n=24)、实验组(n=24)。实验组采用结扎坐骨神经主干的方法构建大鼠坐骨神经损伤模型。测量大鼠疼痛行为学数据,于术后第1,7,14天取材,采用免疫荧光染色技术检测大鼠腰段脊髓背角不同激活状态的小胶质细胞变化;通过qRT-PCR验证不同类型小胶质细胞相关标记物的变化趋势。结果 假手术组大鼠在术后14 d内脊髓背角小胶质细胞形态和数量无明显改变,小胶质细胞标记物也无明显变化。术后1 d,CCI大鼠小胶质细胞形态和数量无明显变化,但促炎型(M1型)标记物增加,提示M1型小胶质细胞活化。术后7 d和14 d,CCI大鼠小胶质细胞数量显著增加,标记物检测显示以M1型活化为主,抑炎型(M2型)小胶质细胞活化不明显。结论 大鼠脊髓背角小胶质细胞在坐骨神经损伤后早期即开始活化,活化持续到至少术后两周,在此期间均以M1型小胶质细胞活化为主。     

胶质细胞源性神经营养因子对局灶性脑缺血后海马齿状回脑电活动的影响

目的：观察胶质细胞源性神经营养因子（GDNF）对局灶性脑缺血大鼠在体海马齿状回脑电活动的影响。方法：制作右侧局灶性脑缺血模型,右侧脑室注射GDNF,通过神经电生理学方法,记录正常组、脑缺血模型组、生理盐水组和GDNF组大鼠局灶性脑缺血90 min后再灌注14 d海马齿状回的脑电活动,分析脑电频谱。结果：免疫组化检测显示,GDNF组各时间点BrdU阳性细胞较脑缺血模型组和生理盐水组增多。大鼠脑电图显示,GDNF组较脑缺血模型组和生理盐水组脑电活动恢复明显,组间比较有显著性差异（均P〈0.05）。结论：GDNF可改善局灶性脑缺血后的脑损伤,促进激活内源性神经干细胞增殖、分化,改善脑缺血后的行为和脑功能。     

人脑出血周围组织星形胶质细胞反应及意义

目的观察人脑出皿周围组织中GFAP在星形胶质细胞的动态表达,探讨脑出血周围组织星形胶质细胞的反应及其意义。方法应用44例因脑出血而死亡的尸检全脑标本,采用HE染色进行形态学观察,免疫组化染色（两步法）标记星形胶质细胞中的胶质纤维酸性蛋白（GFAP）,观察GFAP在星形胶质细胞的表达和变化规律,实验结果应用SPSS11．5软件包进行统计学分析。结果脑出血2h其周围组织中GFAP表达开始增加（P〈0.01）,3～5d GFAP表达明显增强（P〈0.01）,7～15d GFAP表达达到高峰（P〈0．01）,以后有所回落,19d时仍高于对照组（P〈0．01）。结论星形胶质细胞在脑出血周围组织的表达呈抛物线样变化,7～15d反应达高峰;脑出血后星形胶质细胞异常活化且与神经元的存活密切相关,发挥双刃剑作用。     

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.     

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.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Electro-Acupuncture Combined with the Trigger Point Needle-Embedding for Treatment of Primary Trigeminal Neuralgia in 31 Cases

In recent years, the author of this essay has applied electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases, yielding satis- factory results as reported in the following.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Persist in and Carry Forward the Integrative Medical Research——Speech at the 6th National General Congress of Chinese Association of Integrative Medicine by Academician CHEN Zhu,Minister of Ministry of Health

The 6th National General Congress of Chinese Association of Integrative Medicine （CALM） was convened at 19-20, April 2008 in Beijing. Academician CHEN Zhu, the minister of Ministry of Health indicated at the congress that the integration of Chinese and Western medicine is very well in keeping with the situation of our country and the general rule of development in medical science; and as a good integration of Chinese medicine and Western medicine, it is mutually beneficial and advantageous to both of them. Seeing the creativity shown in integrative medical investigation in theoretic and methodological sides, we should and must persist in and develop it.