卡介苗对鼠源性巨噬细胞株释放一氧化氮和IFN-γ的影响

目的:观察卡介苗(BCG)对鼠源性巨噬细胞株RAW264释放的诱导型一氧化氮(NO)合成酶(iNOS)及γ干扰素(IFN-γ)表达的影响.方法:在鼠源性巨噬细胞株RAW264培养液中加入BCG,Griess检测法测定NO释放量,半定量RT-PCR法检测iNOS及IFN-γ的表达.结果:BCG添加后使巨噬细胞NO产生量增加,IFN-γ、iNOS的mRNA表达增强.结论:在巨噬细胞吞噬、杀灭结核分枝杆菌时IFN-γ具有活化巨噬细胞的作用.     

大肠埃希菌感染对U937细胞凋亡的影响及机制探讨

目的探讨大肠埃希菌(E.coli)感染对人巨噬细胞系U937细胞的促凋亡作用及其机制。方法调整U937细胞与E.coli浓度比为120时,放入培养基中培养0、10、20、30、60和90 min,获得感染的U937细胞。采用流式细胞仪检测不同感染时间U937细胞的细胞凋亡率,Western blot法检测U937细胞胞浆内第2线粒体激活剂(Smac)和X连锁凋亡抑制蛋白(XIAP)的表达水平。在E.coli感染前60 min用浓度0、10、20和40μmol.L-1的Em-belin预处理U937细胞,然后在E.coli诱导U937细胞凋亡30 min后收集细胞,用Annexin V FITC/PI双染后用流式细胞仪分析U937细胞的凋亡情况。结果当U937细胞E.coli浓度比为120时,感染0、10、20、30、60和90 min时U937细胞凋亡率分别为(3.02±0.78)%、(6.67±1.34)%、(10.56±1.02)%、(33.92±2.66)%、(46.98±3.12)%和(69.02±4.69)%,呈时间依赖性。U937细胞感染E.coli后发生凋亡,并且凋亡率随着E.coli感染时间的延长而升高。Smac的表达随着E.coli感染时间的延长逐渐升高,XIAP的表达则随着感染时间的延长而逐渐降低,而且不同感染时间组比较差别有统计学意义(P<0.05)。加入XIAP的抑制剂Embelin后,U937细胞的凋亡率随着Embelin的浓度增加而逐渐升高(P<0.05)。结论人巨噬细胞系U937细胞感染E.coli后发生凋亡,其凋亡的发生与影响Smac和XIAP的表达有关。Embelin通过特异性地抑制XIAP表达,增加E.coli诱导的U937细胞凋亡率。     

腹膜透析液中添加黄芪对腹腔巨噬细胞功能的影响

目的:通过在腹膜透析液(PDF)中添加黄芪注射液后观察患者腹腔巨噬细胞功能的变化。　方法:采用自 身对照的方法,在28例连续性不卧床腹膜透析(CAPD)治疗患者的PDF中添加黄芪注射液(20ml/2L)后,比较患 者透出液巨噬细胞活性、吞噬功能及肿瘤坏死因子α(TNF α)和一氧化氮(NO)分泌的变化情况,以及在不同剂量 黄芪注射液的作用下,巨噬细胞活性和细胞因子分泌改变的影响。　结果:用药前后患者透出液中TNF α含量未 出现显著变化,分别为(100±63)ng/Lvs(116±60)ng/L(P=0.192);腹腔巨噬细胞噬菌率分别为(34.8±12.7)% vs(43.4±9.3)%(P<0.01),杀菌率分别为(23.6±7.7)%vs(32.1±10.2)%(P<0.01);巨噬细胞经脂多糖 (LPS)刺激其噻唑蓝(MTT)还原能力检测值(A值)分别为0.156±0.051vs0.230±0.097(P<0.01)。上清液中 NO含量分别为(7.72±2.36)μmol/Lvs(12.22±2.81)μmol/L(P<0.01)。上清液中TNF α含量分别为(1406± 358)ng/Lvs(1925±389)ng/L(P<0.01)。分离的巨噬细胞在不同条件下共同孵育后,PDF组和RPMI1640组巨 噬细胞上清液中TNF α、NO含量和沉渣MTT还原能力较含黄芪注射液各组明显降低(P<0.01);黄芪组上清液中 TNF α、NO含量和沉渣MTT还原能力以1%和2%黄芪组改善较为明显,两组间比较     

分枝杆菌噬菌体D29与巨噬细胞的相互作用及其机制

目的：探讨分枝杆菌噬菌体D29与巨噬细胞的相互作用及其在被吞噬了结核菌的巨噬细胞吞噬后分布于结核菌内外的滴度变化,为探讨D29用于治疗结核病的机制及临床治疗剂量的确定提供参考。方法：将体外培养的巨噬细胞在加入D29不同时间后裂解,测定D29滴度,同时在加入D29 12 h后收集细胞上清液,ELISA法检测一氧化氮(NO)和白细胞介素12(IL-12)水平;将D29加入到已吞噬结核菌的巨噬细胞中,于不同时间采用荧光定量PCR测定D29在结核菌内外分布的滴度。结果：1×108噬菌斑形成单位（PFU）D29加入巨噬细胞后,20 min时有2.5×104PFU被巨噬细胞吞噬,180 min后D29大量失活,190 min时巨噬细胞内已无存活的D29;细胞上清液中NO和IL-12水平与巨噬细胞空白组比较差异无统计学意义(P>0.05),但明显少于LPS阳性对照组(P<0.05);D29与吞噬了结核菌的巨噬细胞作用30 min时,即有D29进入结核菌内,作用40和50 min时,D29在结核菌内外滴度均有一过性升高,此后结核菌内外滴度均下降;作用190和210 min时D29量大幅度减少, 48 h后几乎不能检测到D29。结论:D29被巨噬细胞吞噬后会失活,并且不影响巨噬细胞的免疫功能;D29能够进入被巨噬细胞吞噬的结核菌中,开始时随着时间延长结核菌中的D29数量增多;随后D29数量大量减少,最后消失。     

TNF对结核杆菌感染小鼠的保护作用及机制的研究

目的 :研究肿瘤环死因子 (TNF)对结核杆菌感染小鼠的保护作用及部分作用机制。方法 :小鼠给予不同剂量的基因重组TNF ,2 4h后感染结核杆菌H37Rv株 ,观察半数死亡时间 ,并测定巨噬细胞的吞噬活性、杀菌功能及NK细胞杀伤活性。结果 :高剂量TNF(1× 10 4 U/只 )缩短感染小鼠半数死亡时间 ,适当剂量 (2× 10 3U/只、5 0 0U/只 )能延长感染小鼠半数死亡时间 ;TNF能提高巨噬细胞的吞噬百分率及吞噬指数 ;提高巨噬细胞的杀菌功能 ;能增强NK细胞的杀伤活性。结论 :适当剂量的TNF对结核杆菌感染小鼠具有保护作用 ,其机制可能与增强MΦ的吞噬杀伤活性及NK细胞的杀伤活性有关。     

巨噬细胞对血管内皮细胞增殖、Hoxb2、血管内皮生长因子受体和整合素αvβ3表达的影响

目的建立人巨噬细胞系U937与人脐静脉血管内皮细胞系ECV-304体外共培养模型,以刀豆蛋白A(ConA)作为U937激活剂,研究巨噬细胞调节血管生成的机制.方法实验分4组ECV-304、ConA+ECV-304、U937+ECV-304和ConA+U937+ECV-304.将ECV-304细胞接种,待60%融合时按照不同的分组进行共培养48 h后,流式细胞仪检测细胞周期的变化采用3H-TdR掺入试验检测内皮细胞DNA合成变化;RT-PCR技术检测内皮细胞血管内皮生长因子(VEGF)受体KDR和同源盒(homebox)Hoxb2基因mRNA表达水平的变化;免疫荧光在流式细胞仪上检测整合素受体αvβ3表达的变化.结果ConA激活的U937细胞可使内皮细胞S期、DNA合成明显增加(P＜0.01);ConA刺激的U937细胞使内皮细胞VEGF受体KDR(0.879±0.003)、Hoxb2基因mRNA的表达水平(0.947±0.003)和整合素受体αvβ3的表达水平(10.26±1.73)明显上调(P＜0.01).结论ConA活化的巨噬细胞可通过影响内皮细胞的细胞周期、DNA合成、VEGF受体KDR、Hoxb2及整合素受体αvβ3的表达来促进内皮细胞的增殖、迁移及与基质的黏附,从而调节血管的生成.     

缬沙坦对U937泡沫细胞血管内皮生长因子表达与分泌的影响

【目的】观察缬沙坦对泡沫细胞血管内皮生长因子(VEGF)mRNA表达与蛋白分泌的影响。【方法】以氧化型低密度脂蛋白刺激体外培养的人单核细胞株U937细胞,建立U937泡沫细胞模型。采用不同浓度的缬沙坦(0.01μmol/L、0.1μmol/L、1μmol/L)进行干预。RT-PCR检测泡沫细胞VEGF mRNA表达,ELISA检测泡沫细胞VEGF蛋白的分泌。【结果】U937泡沫细胞VEGF mRNA表达(2.371±0.253)和VEGF蛋白分泌(1804.18±177.59pg/mL)增加,与U937巨噬细胞组(0.954±0.245,716.19±60.82pg/mL)相比,差异有统计学意义(P<0.01)。不同浓度缬沙坦干预后,U937泡沫细胞VEGF mRNA表达水平呈浓度依赖性降低(1.853±0.252、1.265±0.347、1.036±0.156);VEGF蛋白分泌(1625.73±148.54、1388.01±45.82、906.18±78.09pg/mL)也呈浓度依赖性降低,差异有显著性(均P<0.01)。【结论】U937泡沫细胞VEGF mRNA表达、蛋白分泌水平较U937巨噬细胞显著性...     

免疫复合物对单核巨噬细胞的调节作用

目的  研究免疫复合物对单核巨噬细胞的增殖、细胞因子的分泌及吞噬功能的调节作用。方法  免疫复合物组：细胞坏死上清+系统性红斑狼疮（SLE）患者血清;对照组有5组：坏死上清+正常人血清组、坏死上清组、SLE血清组、正常人血清组和培养基组,以上分别作用于U937细胞和U937细胞诱导产生的巨噬细胞。24h后,用CCK 8法检测U937细胞的增殖,RT-PCR法检测U937细胞分泌肿瘤坏死因子α（TNF-α）和B细胞活化因子（BAFF）的变化,中性红比色法检测诱导产生的巨噬细胞的吞噬功能。结果  与对照组相比,免疫复合物能明显促进U937细胞的增殖 （Ｐ＜0.05）,TNF-α 和BAFF 的mRNA表达水平明显增加（Ｐ＜0.05）,并明显抑制巨噬细胞的吞噬功能（Ｐ＜0.05）。结论  在免疫复合物作用下,单核巨噬细胞系统（MPS）处于异常活化状态,使免疫复合物的形成增多而清除减少,从而参与了SLE的发生发展。     

牛磺酸对小鼠腹腔巨噬细胞功能调节作用的体外研究

目的 研究牛磺酸对小鼠腹腔巨噬细胞功能的调节作用。方法 收集小鼠腹腔巨噬细胞,调整细胞浓度为2×10⁵/mL,分为5组,即正常对照组,LPS对照组(LPS终浓度为1 μg/mL)和低、中、高牛磺酸剂量组(牛磺酸的终浓度分别为5、50和500 μg/mL),培养24 h后ELISA法检测细胞上清中IL-12、TNF-α、IL-1、IL-6含量;瑞氏染色法检测并计算巨噬细胞对白色葡萄球菌的吞噬率和吞噬指数、Griess法检测吞噬葡萄球菌后细胞上清中NO的含量。结果 各剂量牛磺酸组的腹腔巨噬细胞TNF-α和IL-6含量均高于正常对照组(P<0.05),高剂量牛磺酸组IL-12的含量高于对照组(P<0.05),各剂量的牛磺酸组IL-1含量与对照组比较没有统计学意义;中、高剂量的牛磺酸组巨噬细胞对葡萄球菌的吞噬率、吞噬指数和上清中NO 的含量均高于正常对照组(P<0.05)。结论 牛磺酸可调节小鼠腹腔巨噬细胞IL-12、TNF-α、IL-6的分泌,但对IL-1的分泌没有调节作用,牛磺酸可促进巨噬细胞对葡萄球菌的吞噬和杀伤功能。     

减毒活菌卡介苗不同时间和剂量干预对哮喘小鼠气道炎症的影响

目的探讨以不同时间和不同剂量给予减毒活菌卡介苗(BCG)对哮喘小鼠气道炎症和气道粘液分泌的影响.方法选择C57BL/6小鼠,以卵白蛋白(OVA)致敏激发建立小鼠哮喘模型,阴性对照组以生理盐水致敏激发.BCG干预组分6组,其中4组分别选择第一次抗原致敏前4 d(组1)、后10 d(组2),最后一次抗原激发后1 h(组3),以及第一次抗原致敏前4 d与后10 d两次(组4),以BCG 105CFUs皮内注射干预;另二组在第一次抗原致敏后10 d分别以BCG 106(组5)、104CFUs(组6)皮内注射干预,在最后一次抗原激发后48 h收集支气管肺泡灌洗液(BALF),计数其中嗜酸性粒细胞(EOS),肺组织PAS染色鉴定粘液分泌.结果OVA致敏激发组BALF中EOS为(41.16±6.42)×104,阴性对照组BALF中几乎见不到EOS.组1、组2和组3 BALF中EOS分别为(10.69±4.35)×104、(7.92±2.30)×104及(17.46±8.00)×104,组5和组6 BALF中EOS分别为(8.71±3.83)×104及(17.19±4.49)×104,均较OVA致敏激发组明显降低(P＜0.05);致敏前、后BCG两次干预(组4)对BALF中EOS[(0.79±0.36)×104]的抑制更为明显,达98%以上.肺组织见OVA致敏激发组气道上皮杯状细胞增生肥大,有大量粘液分泌;阴性对照组未见气道上皮细胞粘液分泌.BCG干预后,气道上皮细胞内粘液分泌减少.结论BCG 105CFUs多次给药对哮喘小鼠气道EOS的抑制作用明显强于单次较大剂量给药,几乎可完全抑制哮喘小鼠气道炎症和粘液分泌;无论致敏前后或激发后BCG均能抑制哮喘小鼠气道炎症,说明BCG对哮喘小鼠具有预防和治疗作用.     

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.