姜酚肟对谷氨酸诱导PC12细胞损伤的保护作用

【目的】探讨姜酚肟对谷氨酸（glutamate，Glu）诱导PC12细胞损伤的保护作用。【方法】体外培养PCI2细胞，建立谷氨酸诱导PC12细胞损伤的模型；采用MTT法检测细胞活力，流式细胞术和Hoechst 33258 DNA染色法检测细胞凋亡。【结果】经5mmol／L的谷氨酸处理24h后，PC12细胞活力比对照组降低，仅为对照组的58．3％。在一定浓度范围内，姜酚肟能保护PC12细胞免受谷氨酸（5mmol／L）的影响，但较高浓度时对细胞有一定的毒性。经不同浓度姜酚肟（3.125、6．25、12．5、25μg／mL）预处理后，细胞活力明显提高，其保护作用随浓度降低而升高，当浓度为6．25μg／mL时效果最好，使PC12细胞的存活率达到75．2％（P〈0．01），浓度为3．125μg／mL时，细胞存活率降为70．2％（P〈0．01）。谷氨酸（5mmol／L）能诱导PCI2细胞凋亡，处理24h后细胞凋亡率为20．1％，经姜酚肟（3．125、6．25、12．5、25μg／mL）预处理后，细胞凋亡率明显降低，分别为3．5％（P〈0．01），2．8％（P〈O．01），9．6％（P〈0．01），17．7％（P〈0．05）。【结论】谷氨酸能诱导PCI2细胞凋亡，较低浓度的姜酚肟能有效保护PC12细胞免受谷氨酸诱导的细胞损伤。其中6．25μg／mL的姜酚肟效果最好。     

人神经干细胞微囊泡对谷氨酸诱导PC12细胞损伤的保护作用

目的: 探究人神经干细胞微囊泡(human neural stem cell microvesicles, hNSC-MVs)对谷氨酸诱导PC12细胞损伤的作用及可能机制。方法: 采用谷氨酸处理大鼠肾上腺髓质嗜铬细胞瘤PC12细胞，建立兴奋性损伤模型；实验分为3组：对照组，PC12细胞未经任何处理；谷氨酸组，谷氨酸处理PC12细胞24 h；hNSC-MVs+谷氨酸组，200 mg/L hNSC-MVs预处理PC12细胞24 h，再加入谷氨酸处理24 h。MTT法检测各组PC12细胞存活率；免疫印迹实验检测细胞凋亡相关蛋白Bax和Bcl-2表达。结果： PC12细胞能内化hNSC-MVs。谷氨酸对PC12细胞具有毒性作用，其半数抑制浓度为25 mmol/L。与谷氨酸组相比，hNSC-MVs+谷氨酸组PC12细胞存活率明显增高(P<0.01)。与对照组相比，谷氨酸组Bax蛋白表达显著升高，Bcl-2蛋白表达显著降低，而与谷氨酸组相比，hNSC-MVs+谷氨酸组Bax蛋白表达明显下调，Bcl-2蛋白表达明显上调(P均<0.01)。结论： hNSC-MVs可能通过抑制谷氨酸诱导的PC12细胞凋亡，从而发挥保护作用。     

硫化氢对化学性缺氧诱导PC12细胞凋亡的影响

【目的】 探讨硫化氢(H2S)对抗二氯化钴(CoCl2)诱导PC12细胞凋亡的作用及其初步机制&#65377;【方法】 应用化学性缺氧模拟剂CoCl2在PC12细胞建立化学性缺氧损伤模型;以硫氢化钠(NaHS)作为H2S的供体;应用CCK-8比色法检测细胞存活率;碘化丙啶(PI)染色流式细胞技术(FCM)检测细胞凋亡率; Hoechst33258 染色检测细胞凋亡的形态学变化; 罗丹明123(Rh123) 染色荧光显微镜照相检测细胞线粒体膜电位(MMP); 2’,7’-二氯荧光黄双乙酸盐(DCFH-DA)染色荧光显微镜照相检测细胞内的活性氧 (ROS)水平&#65377;【结果】 CoCl2引起PC12细胞的存活率显著降低及凋亡率明显增加, 同时引起PC12细胞的MMP 明显降低及ROS 生成显著增加&#65377;在600 μmol/L CoCl2处理PC12细胞前,应用100 ~ 400 μmol/L NaHS 预处理30 min,呈浓度依赖性地阻断CoCl2引起PC12细胞的存活率降低; 200 μmol/L&#65380;400 μmol/L NaHS 预处理能显著地降低600 μmol/L CoCl2 引起PC12 细胞的凋亡率增加并阻断CoCl2 引起的MMP降低及ROS升高&#65377;【结论】 H2S 具有神经细胞保护作用, 能明显地对抗CoCl2诱导的PC12细胞凋亡,此作用可能与其减少ROS生成,提高MMP有关&#65377;     

利福平对鱼藤酮致分化PC12细胞氧化应激的影响

【目的】 探讨利福平对鱼藤酮诱导的分化大鼠嗜铬细胞瘤细胞株(PC12)细胞形态&#65380;活性氧(ROS)&#65380;还原型谷胱甘肽(GSH)及细胞凋亡的影响&#65377;【方法】 利用鱼藤酮诱导分化PC12细胞建立帕金森病体外细胞模型;显微镜下观察细胞形态,酶标仪检测细胞还原型谷胱甘肽,流式细胞仪检测细胞活性氧和凋亡&#65377;【结果】 鱼藤酮组细胞内还原型谷胱甘肽含量低于两对照组,而活性氧含量及凋亡率均高于两对照组;经100&#65380;200和300 μmol/L各浓度利福平预处理后,利福平预防组3组细胞内活性氧含量及凋亡率均低于鱼藤酮组,而细胞内还原型谷胱甘肽含量高于鱼藤酮组,且存在浓度依赖性&#65377;【结论】 利福平可能通过氧化应激途径减轻鱼藤酮诱导的分化PC12细胞的损伤作用,且存在浓度依赖性&#65377;     

ATP敏感钾通道在硫化氢钠对抗β-淀粉样肽 诱导细胞损伤中的作用

 探讨ATP敏感性钾通道 (KATP) 在硫化氢(H2S) 对抗β-淀粉样多肽 (Aβ) 诱导嗜铬细胞瘤细胞(PC12)细胞损伤中的作用?【方法】 应用硫化氢钠 (NaHS) 作为H2S 的供体,碘化丙啶 (PI) 染色流式细胞技术 (FCM) 检测细胞凋亡率,Hoechst染色检测细胞凋亡的形态学变化, 罗丹明123 (Rh123) 染色FCM检测细胞线粒体膜电位 (MMP),双氢罗丹明123 染色FCM检测细胞内活性氧 (ROS) 的含量?【结果】 20 μmol/L和40 μmol/L Aβ25-35能明显地诱导PC12细胞凋亡,增加细胞凋亡率,并能明显地抑制PC12细胞MMP及增加细胞内ROS生成;100 mol/L和200 μmol/L NaHS本身不损伤PC12细胞,但能明显地抑制Aβ25-35的致细胞凋亡作用,降低PC12细胞的凋亡率,并能显著地抑制Aβ25-35引起的MMP降低及胞内ROS水平增多;在应用NaHS与Aβ25-35作用PC12细胞之前30 min,应用KATP抑制剂Glybenclamide (Gly) (10 μmol/L) 对PC12细胞进行预处理,能部分地对抗NaHS的细胞保护作用,使PC12细胞凋亡率增加,MMP降低及ROS生成增多?【结论】 NaHS能明显对抗Aβ25-35对PC12细胞的损伤作用,此细胞保护作用可能与NaHS保护PC12细胞的MMP及抑制胞内ROS生成有关;KATP通道抑制剂Gly能部分地阻断NaHS的细胞保护作用,提示KATP通道开放可能是NaHS对抗Aβ25-35损伤作用的机制之一?     

Par-4反义寡核苷酸拮抗谷氨酸对PC12细胞中ERK1/2的下调作用及其抗凋亡作用

目的研究par-4反义寡核苷酸拮抗谷氨酸对PC12细胞中ERK1/2活性的下调作用及其抗凋亡作用.方法利用脂质体将par-4反义寡核苷酸转染入PC12细胞.谷氨酸诱导PC12细胞凋亡.利用吖啶橙/溴化乙锭荧光染色观察PC12细胞形态,流式细胞分析评价凋亡百分率.Western印迹测定par-4的蛋白质表达量和磷酸化的ERK1/2的蛋白表达量. 结果 (1)谷氨酸诱导PC12细胞中par-4蛋白表达上调,Par-4反义寡核苷酸呈剂量依赖性地拮抗其上调(组间比较,P<0.01).(2)谷氨酸诱导PC12细胞中磷酸化(Thr202/Tyr204)的ERK1/2蛋白水平下调,par-4反义寡核苷酸拮抗其上调(组间比较,P<0.01).(3)Par-4反义寡核苷酸拮抗谷氨酸诱导的PC12细胞凋亡,如予ERK1/2阻断PD98059预处理,则其拮抗作用被下调(组间比较,P<0.05).结论 Par-4反义寡核苷酸拮抗谷氨酸诱导的PC12细胞凋亡,其机制可能与ERK1/2的活化有关.     

莫达非尼抑制过氧化氢诱导的PC12细胞凋亡

目的:研究莫达非尼(modafinil)对过氧化氢诱导的PC12细胞凋亡的抑制作用,并对其机制进行探讨.方法:以噻唑兰(MTT)法测定PC12细胞存活率;用流式细胞术检测PC12细胞凋亡的百分率;测定细胞内罗丹明123(Rhodamine123)的荧光强度,反映细胞线粒体膜电位的改变.结果:莫达非尼能抑制过氧化氢(200 μmol/L)、硝普钠(500μmol/L)和连二亚硫酸钠(2 mmol/L)对PC12细胞的损伤.过氧化氢(200 μmol/L)24h可以诱导PC12细胞凋亡(凋亡率为32.65%).莫达非尼(15,7.5 μmol/L)显著降低凋亡细胞的百分率(凋亡百分率分别为7.95%、15.46%).并且莫达非尼可以抑制过氧化氢引起的线粒体膜电位降低.结论:莫达非尼对PC12细胞有保护作用.能抑制过氧化氢诱导的PC12细胞凋亡,这一作用可能与其抑制过氧化氢引起的线粒体膜电位降低有关.     

隐丹参酮上调GRP78抑制谷氨酸诱导的PC12细胞凋亡

目的：探讨隐丹参酮(Cryptotanshinone, CTS)对谷氨酸(glutamate, Glu)诱导的神经元凋亡的影响及潜在的作用机制。方法：取生长状态良好的PC12细胞,用1 μmol/L Glu处理,Western Blot检测细胞中凋亡相关蛋白Bax、Bcl2的表达水平;以CTS(5 μmol/L或10 μmol/L)预处理PC12细胞12 h后,再用Glu刺激12 h,采用TUNEL染色分析CTS对Glu诱导PC12细胞凋亡的影响;Western Blot检测CTS预处理前后Glu刺激的PC12细胞中糖蛋白调节78(78 ku glucose-regulated protein, GRP78)的蛋白表达水平,并通过sh-RNA干扰细胞内GRP78表达,观察细胞活力的变化。结果：Glu可诱导PC12细胞凋亡,抑制胞内GRP78的表达;经CTS预处理后Glu诱导的神经元凋亡现象得到缓解,GRP78表达增加;干扰PC12细胞内GRP78表达后,可逆转CTS的抗凋亡作用。结论：CTS可通过上调GRP78的表达从而保护PC12细胞抵抗Glu诱导的细胞凋亡。     

BDNF基因修饰淋巴细胞培养上清对PC12细胞氧化应激损伤的保护作用

【目的】研究脑源性神经营养因子基因(brain-derived neurotrophie factor，BDNF gene)修饰淋巴细胞对PC12细胞氧化应激损伤的保护作用。【方法】MTT法检测H2O2和多巴胺(dopamine，DA)对PC12细胞生长的抑制率，Hoechst染色和PI染色流式细胞仪(flow cytometry，FCM)检测H2O2和DA对PC12细胞凋亡的诱导作用。【结果】BDNF基因修饰的大鼠淋巴细胞的培养上清可降低H2O2和DA对PC12细胞生长的抑制率并能抑制H2O2和DA诱导PC12细胞凋亡。【结论】BDNF基因修饰淋巴细胞的培养上清对PC12细胞氧化应激损伤具有保护作用。     

瘦素对缺氧复氧培养L02肝细胞功能和氧化产物的影响

【目的】 观察瘦素对缺氧复氧人肝脏细胞(L02)的肝功能保护和丙二醛(MDA)、超氧化物歧化酶(SOD)的影响。【方法】 将L02细胞分别分为正常对照组、单纯缺氧12 h复氧组和缺氧12 h复氧加不同浓度的瘦素(分别为100、200、400、800和1 600 μg/L)干预组,取细胞上清液检测谷丙转氨酶(ALT)、谷草转氨酶(AST)、MDA的浓度和SOD活性;电镜检测细胞凋亡的形态学改变。 【结果】 ①与正常对照组相比,L02细胞经缺氧12 h复氧培养后,ALT和AST浓度明显升高(P < 0.01),加用不同浓度瘦素干预组ALT和AST浓度较单纯缺氧12 h复氧组下降(P < 0.01);②与正常对照组相比,L02细胞经缺氧12 h复氧培养后,MDA浓度明显升高(P < 0.05),SOD活性下降(P < 0.05),加用不同浓度瘦素干预组MDA浓度较单纯缺氧12 h复氧组下降(P < 0.05);SOD活性上升(P < 0.05);③电镜检测单纯缺氧复氧后细胞呈现明显凋亡形态学改变,加用瘦素100 μg/L处理细胞组细胞仅轻度改变。【结论】 瘦素对缺氧复氧培养导致L02肝细胞的细胞损伤有一定的保护作用;可能与瘦素抑制氧自由基生成,减少脂质过氧化有关。     

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.