双酚 A 对小鼠腹腔巨噬细胞极化影响的体外研究

目的：探讨体外实验条件下,双酚 A（BPA）对小鼠腹腔巨噬细胞极化的影响。方法提取 C57BL/6J 雄性小鼠腹腔巨噬细胞,用（10 ng/ ml）干扰素-γ（IFN-γ）和（500 ng/ ml）细菌脂多糖（LPS）诱导其向 M1型极化;用（10 ng/ ml）白介素-4（IL-4）诱导其向 M2型极化;同时加入0.1、1、10μmol/ L BPA 处理细胞,并设立空白对照组和溶剂对照组。流式细胞术检测 M1型和 M2型巨噬细胞比例,ELISA 法检测 M1型和M2型巨噬细胞各自分泌的诱导型一氧化氮合酶（ iNOS）和精氨酸酶（Arg-1）活性。结果不同浓度 BPA 促进 IFN-γ和LPS 诱导的小鼠腹腔巨噬细胞向 M1型极化,1、10μmol/ L BPA 组 M1型巨噬细胞比例较空白对照组分别升高11.3%和17.4%,M1型巨噬细胞分泌的 iNOS 活性分别升高1.95和2.29倍,差异均有统计学意义。不同浓度 BPA 抑制 IL-4诱导的小鼠腹腔巨噬细胞向 M2型极化,1、10μmol/ L BPA组 M2型巨噬细胞比例分别较空白对照组降低9.0%和14.3%,M2型巨噬细胞分泌的 Arg-1活性分别下降0.37和0.49倍,差异均有统计学意义。结论体外条件下,低剂量BPA 促进小鼠腹腔巨噬细胞向 M1型极化,抑制其向 M2型极化。     

Hippo信号通路关键蛋白YAP调控巨噬细胞极化的分子机制研究

目的:探讨Hippo信号通路关键蛋白YAP调控巨噬细胞极化的分子机制.方法:单核细胞系THP-1细胞经佛波酯诱导成为巨噬细胞作为对照组,经脂多糖诱导为M1型巨噬细胞组,经白细胞介素-4诱导为M2型巨噬细胞组,取对数生长期的M1型巨噬细胞分别转染YAP表达质粒、对照质粒及空载体,收集各组细胞,采用流式细胞术检测M1型和M2型巨噬细胞标志物水平,酶联免疫吸附法检测M1型和M2型巨噬细胞特异性分泌因子水平,实时定量PCR检测mRNA表达水平,Western blot检测蛋白表达.结果:流式细胞术检测结果显示,与对照组细胞相比,诱导的CD86+(M1型巨噬细胞标志物)和CD206+(M2型巨噬细胞标志物)细胞比例增加(P<0.05);酶联免疫吸附法检测结果显示,M1型巨噬细胞组培养上清液中TNF-α水平高于对照组和M2型巨噬细胞组(P<0.05),M2型巨噬细胞组培养上清液中TGF-β水平高于对照组和M1型巨噬细胞组(P<0.05),提示M1型和M2型巨噬细胞诱导成功.实时定量PCR检测结果显示,YAP在M1型巨噬细胞中的表达水平低于M2型巨噬细胞(P<0.05);Western blot检测结果显示,YAP在M1型巨噬细胞中的表达水平低于M2型巨噬细胞(P<0.05),提示YAP可能与M2型巨噬细胞的表型维持有关.实时定量PCR检测结果显示,与对照质粒组相比,YAP表达质粒组中,M1型巨噬细胞标志物IL-1βmRNA表达水平降低(P<0.05),M2型巨噬细胞标志物CCL22 mRNA表达水平升高(P<0.05);Western blot检测结果显示,与对照质粒组相比,YAP表达质粒组中,M1型巨噬细胞标志物IL-1β 蛋白表达水平降低(P<0.05),M2型巨噬细胞标志物CCL22蛋白表达水平升高(P<0.05),提示YAP可能抑制M1型巨噬细胞的形成和促进M2表型形成.结论:Hippo信号通路关键蛋白YAP表达高低可调控巨噬细胞极化,YAP表达升高可抑制M1型巨噬细胞的形成和促进M2表型形成.     

巨噬细胞极化为M1型或M2型对铁代谢的影响

目的探讨巨噬细胞极化形成M1型或M2型对铁代谢的影响。方法应用脂多糖（LPS）20ng/mL或IL-410ng/mL诱导RAW264.7巨噬细胞极化,电子显微镜下观察处理后的巨噬细胞极化形态特征,采用免疫荧光染色法及Westernblot法检测M1型标志物TNF-α和一氧化氮合酶（iNOS）及M2型标志物精氨酸酶1（Arg-1）的表达情况;LPS或IL-4处理后的巨噬细胞与红细胞及铁剂共培养,通过瑞氏染色和铁染色,评价其铁摄入功能;ELISA法检测LPS或IL-4处理后巨噬细胞内及培养液中铁蛋白含量,以评价其铁储存功能。结果LPS处理后巨噬细胞由圆形向多形性转化;同时,TNF-α和iNOS的表达水平明显增加;可见吞噬红细胞现象,胞质中吞噬的铁颗粒明显增多。IL-4处理后巨噬细胞形态也呈多形性改变,同时Arg-1表达水平增加,但未见吞噬红细胞现象,且胞质内吞噬的铁颗粒较少。此外,LPS处理后巨噬细胞胞内铁蛋白含量较IL-4处理后明显增多。结论LPS与IL-4可诱导巨噬细胞形成不同极化亚型,其铁代谢存在明显差异。LPS处理的M1型巨噬细胞对铁的摄入及储存较IL-4处理的M2亚型巨噬细胞增加,有固铁优势。     

Pyrin在巨噬细胞M1型极化中的作用研究

目的：探讨Pyrin在巨噬细胞M1型极化中的作用。方法：培养人单核细胞株THP-1细胞,提取人外周血单核细胞（peripheral blood mononuclear cell,PBMC）、小鼠骨髓来源巨噬细胞（bone marrow-derived macrophage,BMDM）。随机分为对照组和实验组,实验组用脂多糖（lipopolysaccharide,LPS,100 ng/mL）复合干扰素-γ（interferon-γ,IFN-γ,20 ng/mL）干预（THP-1 18 h,PBMC、BMDM 24 h）构建巨噬细胞M1型极化模型。qRT-PCR、Western blot和流式细胞术检测2组M1型极化相关标志物,同时利用qRT-PCR和Western blot检测MEFV基因和Pyrin蛋白表达水平。结果：与对照组相比,实验组在LPS+IFN-γ刺激后,qRT-PCR检测THP-1细胞M1型极化标志物：肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）（t=-4.360,P=0.007）、白细胞介素-1β（interleukin-1β,IL-1β）（t=-8.329,P=0.000）、白细胞介素-6（interleukin-6,IL-6）（t=-2.924,P=0.033）、CD14（t=-2.858,P=0.035）、CD80（t=-3.433,P=0.019）,PBMC细胞M1型极化标志物：TNF-α（t=-2.893,P=0.034）、IL-1β（t=-3.606,P=0.015）、IL-6（t=-2.895,P=0.034）、CD14（t=-2.645,P=0.046）、CD80（t=-3.648,P=0.015）,BMDM细胞M1型极化标志物：TNF-α（t=-6.123,P=0.002）、IL-1β（t=-2.697,P=0.043）、单核细胞趋化蛋白-1（monocyte chemotactic protein 1,MCP-1）（t=-4.335,P=0.007）、诱导型一氧化氮合酶（inducible nitric oxide synthase,iNOS）（t=-3.607,P=0.015）均明显增加,Western blot检测THP-1细胞M1型极化标志物：TNF-α（t=-3.827,P=0.031）、IL-1β（t=-4.189,P=0.025）、IL-6（t=-5.345,P=0.002）均明显增加,流式细胞术检测THP-1细胞M1型极化标志物HLA-DR（t=-3.270,P=0.017）、BMDM细胞M1型极化标志物F4/80+CD11c+（t=-3.833,P=0.009）均明显增加,提示M1型极化模型构建成功;在构建成功的M1型极化模型中,qRT-PCR检测THP-1细胞、PBMC细胞和BMDM细胞MEFV基因表达水平较对照组明显增加（t=-3.226,P=0.023;t=-2.989,P=0.030;t=-2.891,P=0.034）,Western blot检测THP-1细胞Pyrin蛋白表达水平较对照组明显增加（t=-8.591,P=0.000）。结论：巨噬细胞M1型极化中Pyrin表达增加,Pyrin可能与巨噬细胞M1型极化相关。     

羟氯喹介导NF-κB/NLRP3通路抑制M1型巨噬细胞极化缓解小鼠肠道炎症

目的：探讨羟氯喹（hydroxychloroquine,HCQ）对葡聚糖硫酸钠（dextran sulfate sodium salt,DSS）诱导的结肠炎模型鼠肠道炎症的影响及其M1型巨噬细胞极化机制。方法：C57BL/6小鼠随机分3组：对照组（正常饮水+200μL纯水每日灌胃）、DSS组（自由饮用3.5%DSS溶液+200μL纯水每日灌胃）、DSS+HCQ组[自由饮用3.5%DSS+200μL HCQ溶液（60 mg/kg）每日灌胃]。造模期间观察小鼠粪便性状,记录体重、疾病活动指数（disease activity index,DAI）评分。造模后测量小鼠结肠长度、结肠组织HE染色行组织病理学评分;提取结肠组织固有层单个核细胞,流式细胞术检测M1型巨噬细胞比例。体外提取并诱导分化小鼠骨髓来源的巨噬细胞,HCQ处理后,流式细胞术检测M1型巨噬细胞比例;蛋白质印迹法检测p-STAT1、IRF5、NF-κB/p65等表达水平。结果：HCQ可以减轻DSS诱导的小鼠肠道炎症并减少结肠中M1型巨噬细胞比例。细胞实验显示HCQ可抑制M1型巨噬细胞极化并同时抑制NF-κB信号通路及其下游NOD样...     

人骨髓间充质干细胞分泌的外泌体调控恶性胶质瘤相关巨噬细胞的极化

  目的  探究人骨髓间充质干细胞分泌的外泌体对肿瘤相关巨噬细胞极化的影响，并初步探讨其对胶质瘤发生发展的影响。  方法  利用试剂盒提取人骨髓间充质干细胞分泌的外泌体，通过扫描电镜及Western blot 鉴定外泌体。将胶质瘤细胞分为SHG449:SHG449细胞不做特殊处理；SHG449+M0组:SHG449细胞与M0型巨噬细胞共培养；SHG449+M0+exosome组:SHG449细胞与M0型巨噬细胞共培养后，加入标记有PKH-67的外泌体使其进入M0型巨噬细胞。  结果  （1）人骨髓间充质干细胞分泌的外泌体成功提取，且进入M0型巨噬细胞；（2）外泌体进入胶质瘤细胞SHG449诱导极化的巨噬细胞后细胞形态转化，且M2型巨噬细胞活化标志物Arginase、CD206以及CCL22，TGF-β表达下降（Arginase:P < 0.01，CD206:P < 0.05， CCL22:P < 0.05，TGF-β:P < 0.01），M1型活化标志物iNOS，CD68以及IL-1β，TNF-α表达上升（iNOS:P < 0.01，CD68:P < 0.01，L-1β:P < 0.000 1，TNF-α:P < 0.001）；（3）被外泌体诱导极化的巨噬细胞导致胶质瘤细胞增殖能力下降（P < 0.05），凋亡能力上升（P < 0.000 1）。  结论  人骨髓间质细胞分泌的外泌体可抑制肿瘤相关巨噬细胞向M2表型转化，并诱导其向M1表型转化，调节肿瘤免疫微环境，从而达到抑癌的作用。     

心肌缺氧状态下间充质干细胞对巨噬细胞 M2 极化的影响及其机制研究

目的 探讨心肌缺氧状态下间充质干细胞（MSC）对巨噬细胞极化的影响,并揭示其调控巨噬 细胞极化的机制。方法 在M0 型巨噬细胞培养液中加入无糖无氧培养的心肌细胞上清液,与MSC 共培养。 采用流式细胞术、Western blotting 及逆转录聚合酶链反应检测M1/M2 型巨噬细胞生物学标志,以判断MSC 在模拟心肌缺氧状态下对巨噬细胞极化的影响,并观察MSC 对巨噬细胞髓样分化因子88 及其上游TLR2、 TLR4 受体表达的影响,以及其下游的细胞核转录因子-κB（NF-κB）和丝裂原活化蛋白激酶（MAPK） 信号通路磷酸化的水平。结果 与M0 组比较,模拟心肌缺氧状态下培养的M0 巨噬细胞成功诱导M1 极化 （P <0.05）;与MSC 共培养促进M1 向M2 极化（P <0.05）。同等条件下加入不同浓度髓样分化因子88 抑制剂后, 与M0+OGD 组比较,M2 型巨噬细胞比例随着ST2825 剂量的增加逐渐升高（P <0.05）。与M0+OGD 组比较, M0+OGD+MSC 组白细胞介素10、转化生成因子-β1 表达水平升高（P <0.05）。Western blotting 检测显示 MSC 共培养后,巨噬细胞TLR2、TLR4 及髓样分化因子88 表达下调,其下游的NF-κB 和MAPK 信号通 路标志分子P65、IKKAα/β、JNK1/2、P38、ERK1/2 等磷酸化水平降低。结论 在心肌缺氧状态下,MSC 促进巨噬细胞M2 极化。其机制之一可能是MSC 下调巨噬细胞TLR2、TLR4 及髓样分化因子88 表达,抑制 TLR2/4- 髓样分化因子88 信号及其下游的NF-κB 和MAPK 通路。     

C型凝集素Dectin-2对动脉粥样硬化M2型巨噬细胞活化和凋亡的作用及机制研究

目的 探究Dectin-2（树突状细胞相关凝集素-2）对动脉粥样硬化进程中巨噬细胞M2型活化和凋亡的功能及机制研究。方法 利用IL-4（白介素4）刺激巨噬细胞M2型活化,WB（蛋白免疫印迹）检测Dectin-2的表达;巨噬细胞敲降Dectin-2,Q-PCR（荧光定量PCR）检测巨噬细胞M2活化标志物Arg1（精氨酸酶1）、FIZZ1（抵抗素样分子α）、Ym1（几丁质酶3）的表达;ox-LDL（氧化修饰低密度脂蛋白）刺激巨噬细胞,流式细胞法检测巨噬细胞凋亡水平;IL-4刺激巨噬细胞0 min、5 min、30 min、60 min检测STAT6（信号传导及转录激活蛋白6）磷酸化变化。结果 IL-4刺激巨噬细胞Dectin-2表达上调,敲降Dectin-2抑制IL-4诱导的巨噬细胞M2型活化标志物Arg1、FIZZ1、Ym1的表达,同时抑制ox-LDL诱导的巨噬细胞凋亡。WB结果显示敲降Dectin-2明显抑制IL-4诱导的STAT6磷酸化水平升高。结论 IL-4刺激巨噬细胞Dectin-2表达上调,抑制Dectin-2表达可以明显抑制巨噬细胞M2型活化以及ox-LDL诱导的凋亡,其作用机制可能是通过调控STAT6的磷酸化水平。     

SOCS3调控JAK/STAT3通路对PD-1/PD-L1抑制剂所致小鼠心脏毒性的影响

目的 探讨细胞因子信号转导抑制因子3（SOCS3）通过调控酪氨酸激酶/转录激活因子3（JAK/STAT3）通路影响巨噬细胞M1型极化的作用机制,及其对程序性死亡受体1/程序性死亡配体1（PD-1/PD-L1）抑制剂所致小鼠心脏毒性的影响。方法 将RAW 264.7细胞分为对照组、LPS组、pc-NC组、pc-SOCS3组、si-NC组、si-SOCS3组及抑制剂组,细胞转染相应质粒并使用100 ng/mL LPS干预;50只SPF级6周龄BALB/c小鼠分为正常组、模型组、NC组、SOCS3组及抑制剂组,每组10只,除正常组外,其余各组小鼠通过腹腔注射PD-1/PD-L1抑制剂BMS-1 10 mg/kg建立PD-1/PD-L1抑制剂诱导的小鼠心脏毒性模型,并通过尾静脉注射相应质粒或腹腔注射相应药物。HE染色观察小鼠心脏组织病理;ELISA法检测细胞上清和小鼠血清IL-10、TNF-α和IL-1β水平;Western blot检测细胞SOCS3、CD86、CD80以及JAK/STAT3信号通路相关蛋白表达。结果 细胞过表达SOCS3会促进巨噬细胞细胞炎症因子水平和M1型极化,并抑制JAK/STAT3通路相关蛋白表达;细胞SOCS3低表达后,细胞炎症因子水平降低,并抑制巨噬细胞M1型极化,激活JAK/STAT3通路（P＜0.05）;BMS-1干预后小鼠心脏组织损伤严重,心脏指数、血清炎症水平及CD86、CD80蛋白表达明显升高,JAK/STAT3通路相关蛋白表达明显降低（P＜0.05）;过表达SOCS3组小鼠心脏损伤减轻,心脏指数、血清炎症因子及心脏CD86、CD80蛋白表达明显降低,JAK/STAT3通路相关蛋白表达明显升高（P＜0.05）;JAK/STAT3通路抑制剂AG490逆转了低表达SOCS3对PD-1/PD-L1抑制剂诱导小鼠心脏损伤减轻作用和M1型巨噬细胞极化作用。结论 低表达SOCS3可通过激活JAK/STAT3通路抑制巨噬细胞M1型极化并减轻PD-1/PD-L1抑制剂所致小鼠心脏毒性     

rL-hIFN-λ1对THP-1源巨噬细胞极化作用的影响

［摘要］目的: 探讨表达IFN-λ1的重组新城疫病毒rL-hIFN-λ1对巨噬细胞极化的影响。方法: 分别用佛波酯、IFN-γ、脂多糖、IL-4和rL hIFN-λ1刺激人外周血THP-1单核细胞系,构建THP-1 M0、THP-1 M1、THP-1 M2、THP-1 rL-hIFN-λ1型巨噬细胞模型。显微镜下观察M0、M1、M2型巨噬细胞形态特点;实时荧光定量PCR(qRT-PCR)检测各组巨噬细胞 M1/M2相关基因表达;免疫荧光检测细胞表面分子CD86、CD163;ELISA检测分泌因子IL-2、IL-13、IL-10及肿瘤坏死因子-α(TNF-α)的表达水平;并采用免疫组织化学法检测不同临床分期肺癌组织中巨噬细胞浸润表型。结果： 诱导得到的M0、M1、M2型巨噬细胞形态差异显著。rL-hIFN-λ1诱导的巨噬细胞中,M1型巨噬细胞标志物表达明显升高,M2型巨噬细胞标志物表达显著降低,同时其可促进M1型巨噬细胞因子释放,抑制M2型巨噬细胞因子释放。随着肺癌临床分期进展,M1型巨噬细胞浸润逐渐减少。 结论： rL-hIFN-λ1可诱导巨噬细胞向M1型巨噬细胞极化。     

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.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

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.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.