急性脑缺血模型大鼠的血浆蛋白质组学研究

目的 探讨急性脑缺血模型大鼠与正常对照之间的血浆蛋白质变化.方法 用线拴法制备脑缺血大鼠模型,缺血2h后采集模型组与空白对照组的大鼠血浆,利用双向电泳、图像分析、质谱鉴定蛋白质组学技术进行分析.结果 鉴定出C-反应蛋白、触珠蛋白、血清淀粉样蛋白P在脑缺血大鼠血浆中表达上调,血清前白蛋白表达下调.结论 所鉴定蛋白可能与脑缺血有一定相关性.     

正常与脑缺血大鼠脑皮质线粒体的比较蛋白质组学研究

目的 采用比较蛋白质组学方法观察正常与脑缺血大鼠脑皮质线粒体蛋白表达差异.方法 SD大鼠按随机数字表法分为正常组和缺血组,采用改进的栓线法制备大脑中动脉栓塞(middle cerebral antery occlusion,MCAO)模型,分离大鼠缺血及正常大脑皮层,纯化线粒体蛋白进行二维电泳和凝胶图像分析,对差异蛋白质用HPLC-Chip/MS纳流液质联用技术进行序列分析,经Spectrum Mill搜索NCBInr数据库后鉴定蛋白质.结果 正常与脑缺血大鼠脑皮层线粒体蛋白表达存在显著差异,这些差异蛋白经鉴定,主要为TUC-4b、肌酸激酶同工酶、HS1结合蛋白、线粒体核糖体蛋白S27、异柠檬酸脱氢酶、丙酮酸脱氢酶、二氢硫辛酸脱氢酶、泛醌-细胞色素C还原酶、琥珀酸辅酶A还原酶、细胞色素氧化酶、F1-ATP beta链、热休克蛋白70、热休克蛋白60、脂酰辅酶A脱氧酶等差异线粒体蛋白.结论 脑缺血可以导致皮层线粒体相关蛋白表达改变,提示其作用可能与线粒体能量代谢、凋亡有关.     

利多卡因对脑缺血再灌注损伤大鼠海马组织细胞间NF-κB表达的影响

目的观察利多卡因对大鼠脑缺血再灌注损伤后海马组织细胞间NF-κB基因表达的影响,探讨利多卡因脑保护作用的机制。方法雄性SD大鼠32只,随机分为假手术组(A组)、缺血再灌注组(B组)、利多卡因大剂量组(C组)和利多卡因小剂量组(D组),缺血前10min腹腔注射。脑缺血10min再灌注24h时,断头处死大鼠。用RT-PCR技术检测海马组织NF-κB的表达,用免疫组织化学、蛋白印记(westernblot)方法检测NF-κB蛋白表达情况。结果脑缺血再灌注后海马组织NF-κBmRNA及蛋白表达水平增高,利多卡因可下调NF-κB表达,缺血再灌注后,海马区神经元细胞出现明显坏死,利多卡因可减轻海马区神经元损伤。结论利多卡因可能通过抑制NF-κB基因表达而对脑缺血再灌注损伤起到一定的保护作用。     

局灶性缺血性脑卒中大鼠海马Notch信号通路的动态表达

目的 观察局灶性缺血性脑卒中大鼠海马Notch信号通路的表达变化,初步探讨该通路在局灶性缺血性脑卒中后神经再生的调控作用.方法 大脑中动脉阻塞(MCAO)法建立大鼠局灶性脑缺血模型,通过Western印迹法和实时RT-PCR法分别于卒中后19 d和28 d检测各实验组及对照组大鼠海马Notch1及其配体Jagged1的蛋白、基因表达.结果 卒中后19 d,缺血组Notch1蛋白和基因表达均分别高于对照组(均P<0.05),缺血组28 d的Notch1蛋白和基因表达显著低于19 d的蛋白和基因表达(均P<0.01).缺血组19 d和28 d的Jagged1蛋白表达(P<0.05)和基因表达(P<0.05)高于对照组,但28 d的Jagged.蛋白和基因表达与19 d相比差异无统计学意义(均P>0.05).结论 局灶性缺血性卒中大鼠Notch信号通路呈现动态表达变化,提示该通路可能参与了对卒中后内源性神经前体细胞增殖、分化的调控.     

西维来司钠对脑缺血/再灌注大鼠全脑蛋白质组的影响

目的:研究西维来司钠(Sivelestat sodium hydrate,ONO-5046)对脑缺血/再灌注(Cerebral ischemia/reperfusion,CI/R)大鼠全脑蛋白质表达的影响.方法:大鼠随机分为假手术组、CUR组和CI/R+ONO-5046组(6 mg/kg).采用四血管阻断法制备CUR模型,缺血15 min,再灌注24 h.提取脑组织总蛋白进行双向电泳,对差异蛋白点用HPLC-Chip-MS/MS纳流液质联用技术进行序列分析,经Spectrum Mill搜索NCBInr数据库后鉴定蛋白质.结果:成功获得了分辨率和重复性好的大鼠脑组织蛋白2-DE图谱,经质谱鉴定最终获得了巨噬细胞炎性蛋白1-beta、白细胞弹性蛋白酶A、β2-巨球蛋白、谷胱甘肽S-转移酶omega-1、谷氨酸脱氢酶-1、细胞色素C还原酶铁硫亚单位、硫氧还蛋白过氧化物酶1等19个差异蛋白点相关信息.结论:大鼠脑缺血后脑组织蛋白质表达存在明显差异,西维来司钠可通过调节有关蛋白质的表达,参与大鼠脑缺血神经元保护作用.     

利多卡因对脑缺血再灌注损伤大鼠海马组织细胞间黏附分子-1表达的影响

目的观察利多卡因对大鼠脑缺血再灌注损伤后海马组织细胞间黏附分子-1(ICAM-1)基因表达的影响,探讨利多卡因脑保护作用的机制。方法雄性SD大鼠32只,随机分为假手术组(A组)、缺血再灌注组(B组)、利多卡因大剂量组(C组)和利多卡因小剂量组(D组),缺血前10 min腹腔注射。脑缺血10 min再灌注24 h时,断头处死大鼠。用RT-PCR技术检测海马组织ICAM-1的表达,用免疫组织化学、蛋白印记(Western blot)方法检测ICAM-1蛋白表达情况。结果脑缺血再灌注后海马组织ICAM-1 mRNA及蛋白表达水平增高,利多卡因可下调ICAM-1表达,缺血再灌注后,海马区神经元细胞出现明显坏死,利多卡因可减轻海马区神经元损伤。结论利多卡因可能通过抑制ICAM-1表达而对脑缺血再灌注损伤起到一定的保护作用。     

大鼠脑缺血后皮质PSD-93基因表达的动态变化

目的：观察突触后密度（PSD）-93基因在大鼠局灶性脑缺血再灌注皮质表达的变化。探讨其在缺血再灌注损伤中的病理作用。方法：线栓法建立大鼠大脑中动脉闭塞再灌注模型，缺血2h后，分别再灌注6、12和24h，应用RT—PCR、Western blot技术检测缺血再灌注后皮质PSD-93基因mRNA和蛋白的表达。结果：缺血再灌注的非缺血侧大脑皮质PSD-93 mRNA和蛋白的表达与对照组相比均无明显变化；缺血侧皮质PSD-93 mRNA表达明显高于未缺血侧，12、24h组与未缺血侧比较具有显著性差异（均P〈0.05），并呈时间依赖性；再灌注6h后PSD-93蛋白表达升高，再灌注12h达高峰，12h组与未缺血侧比较具有显著性差异（P〈0．05）。结论：脑缺血再灌注后皮质PSD-93基因表达增高，推测PSD-93参与介导缺血性脑损伤。     

低氧预处理对全脑缺血大鼠脑内cPKC和cPKCα蛋白表达的影响

目的:探讨低氧预处理对全脑缺血大鼠脑内cPKC和cPKCα蛋白表达的影响及其意义。方法:将24只Wistar雄性大鼠,随机分成正常对照组和缺血再灌注组及低氧预处理+缺血再灌注组,利用免疫组化的方法观察cPKC和cPKCα蛋白表达。结果:脑缺血后缺血组织周围cPKC和cPKCα蛋白表达均有增高但是在低氧预处理组有明显降低的表现尤其以cPKC为主。讨论:脑缺血后缺血组织周围cPKC和cPKCα蛋白表达上调可能与神经细胞凋亡有关,提示细胞信号转导系统异常可能参与了脑缺血后神经细胞损伤的病理生理过程。这可能是低氧预处理可以降低脑缺血后脑组织的进一步脑损害的作用机制。     

大脑中动脉阻塞大鼠脑内脑源性神经营养因子蛋白表达变化

目的观察大脑中动脉阻塞大鼠脑内脑源性神经营养因子蛋白的表达变化。方法采用大鼠大脑中动脉阻塞再灌流模型，应用免疫组织化学方法测定脑缺血后3、7以及30d脑内BDNF蛋白的表达；结果缺血组大鼠脑内皮质、海马、缺血灶周围等处有BDNF的表达。脑缺血后3d上述部住BDNF。的表达增高，7d后其表达水平继续增强达高峰，30d后BDNF阳性细胞数量及表达水平降低。阳性细胞主要表达于神经元，其中在7d、30d2个时间点上，两组阳性细胞计数比较差异有显著性（P〈0.01）。结论脑缺血后BDNF蛋白的表达呈时段性增高。     

颈内动脉注射质粒pLXSN-bcl-2对大鼠局灶性脑缺血后热休克蛋白70表达的影响

目的观察bcl-2基因的过度表达在局灶性脑缺血中的脑保护作用,及能否引起热休克蛋白70(HSP70)表达的变化。方法参照改良的Longa栓线法构建大鼠局灶性脑缺血模型,缺血2h后将栓线拔出,经颈内动脉向大鼠脑内分别注人质粒pLXSN-bcl-2、空质粒pLXSN和生理盐水各260μl。存活的各组大鼠分别再灌注3,24,72h,断头取大脑。用免疫组织化学方法观察Bcl-2蛋白和HSP70蛋白表达情况。结果再灌注3h大鼠脑内即可见Bcl-2蛋白,HSP70表达,表达逐渐增加,二者均在再灌注24h达到高峰,再灌注72h表达降低,二者均以缺血区表达最强,但Bcl-2组两种蛋白的表达在各时间点均较对照组(空质粒组和生理盐水组)强,而对照组之间两种蛋白的表达在各个时间点均无差异。结论大鼠颈内动脉注射质粒pLXSN-bcl-2后,局灶性脑缺血大鼠脑组织中Bcl-2蛋白的广泛表达促进了HSP70基因的表达,保护了缺血脑组织。     

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.