降钙素基因相关肽抑制白细胞介素-1介导骨的吸收作用

目的　了解白细胞介素 - 1β(IL - 1β)在破骨细胞性骨吸收中的刺激作用 ,并评价降钙素基因相关肽 (CGRP)的拮抗效果 .方法　将新生大鼠破骨细胞和成骨细胞混合培养 ,接种于预置象牙片的培养板中 .2 4 h后培养液中分别加入 IL-1β和不同浓度的 CGRP.继续培养 4 8h,然后取出象牙片 ,超声处理后行甲苯胺蓝染色 ,光镜下观察骨吸收陷窝数目并计算其总面积 .结果　 IL - 1β明显增加象牙片上吸收陷窝的数目 (37.2± 8.2 vs 2 2 .4± 5 .7) per slice (P <0 .0 1)和面积(μm2 ,976 1± 2 775 vs4 2 2 4± 1381,P<0 .0 1) ,而 CGRP则能够显著抑制 IL- 1β的刺激作用 (P<0 .0 5或 P<0 .0 1) ,且抑制作用呈剂量依赖性 (r1 =- 0 .5 6 ,P<0 .0 1;r2 =- 0 .4 9,P<0 .0 5 ) .结论　 CGRP可直接作用于破骨细胞并抑制其活化 ,并调节成骨细胞相关因子的释放而间接影响破骨细胞功能 .     

牙周炎动物模型的研究

目的：建立一种典型、稳定的牙周炎动物模型。方法：将24只健康大耳白兔随机分为4组，磨牙颈部结扎钢丝，在兔的牙周组织注射IL－1－β、TNF－α，以生理盐水为对照，分不同时期取材做组织学观察和评价。结果：实验组15d后牙龈组织血管扩张充血，牙槽骨改建活跃，有大量的骨吸收陷窝及破骨细胞，并有新骨形成和成骨细胞出现，30d后仅表现为大量的骨吸收陷窝及破骨细胞，无成骨现象，且以联合应用IL－1－β、TNF-α组最为显著。结论：IL－1－β、TNF－α在建立牙周炎动物模型中有重要作用。     

组织块法与传统破骨细胞培养方法对骨吸收功能的比较研究

目的：比较组织块法与传统破骨细胞培养方法对破骨细胞骨吸收功能的影响。方法：用组织块法与传统破骨细胞培养方法进行破骨细胞培养,抗酒石酸酸性磷酸酶（tartrate-resistant acid phosphatase,TRAP）染色鉴定破骨细胞,超声去除骨片上细胞后,骨片经1%甲苯胺蓝染色,光镜下观察骨片上吸收陷窝。根据骨片上陷窝数目、面积以及深度定量比较两组破骨细胞骨吸收功能。结果：吸收陷窝经甲苯胺蓝染色后光镜下可清晰识别,组织块法在骨片上的吸收陷窝数目比传统法明显少,而且骨片上陷窝面积及深度也比传统法小而浅（P〈0.05）。结论：组织块法培养的破骨细胞无论从数量上还是从功能上均没有传统法培养的好。数量大的成骨细胞对共培养破骨细胞骨吸收功能起到负面作用。     

饲补肾中药大鼠血清对成骨细胞-破骨细胞共育系中破骨细胞功能的影响

目的探讨喂饲补肾中药大鼠血清在成骨细胞-破骨细胞培养体系中对破骨细胞骨吸收功能的影响。方法取SD大鼠的胎鼠颅骨与四肢骨分别分离、培养成骨细胞和破骨细胞,利用共育系使二者生活在同一环境中,施加高、中、低剂量的饲补肾中药大鼠血清,并与对照组及单独培养的破骨细胞进行比较,酶动力学法测定培养上清中抗酒石酸酸性磷酸酶(TRACP)的活性,利用甲苯胺蓝对骨吸收陷窝染色并在Leica Quantimet 500图像分析仪下测定骨吸收陷窝的面积和数目。结果共育系中对TRACP活力的抑制及骨陷窝面积与数目的减少以中剂量组作用最为明显(与对照组比较P<0.01),单独培养3个剂量组与对照组比较均有显著性差异(P<0.05),以中剂量组作用最为明显(P<0.01)。结论饲补肾中药大鼠血清可以抑制共育系中破骨细胞的骨吸收功能。     

血小板衍生生长因子-BB在成骨细胞-破骨细胞培养体系中的生物学作用

目的　探讨血小板衍生生长因子（PDGF）-BB在成骨细胞-破骨细胞培养体系中对破骨细胞骨吸收功能的影响。方法　体外分离、培养成人成骨细胞和破骨细胞,利用复合培养系统使二者生活在同一环境中;在复合培养体系中施加不同浓度的PDGF-BB或者PDGF-BB+L-单甲基精氨酸(L-NMMA);酶动力学法测定培养上清中抗酒石酸酸性磷酸酶（TRAP）的活性;利用甲苯胺蓝对骨吸收陷窝染色并在图像分析仪下测定骨吸收陷窝的面积和数目。结果　在PDGF-BB单独作用下,复合培养体系中TRAP的活性不随POGF-BB的浓度发生明显变化（P>0.05）,骨吸收陷窝的面积和数目PDGF-BB均与对照组比较,无明显变化（P>0.05）;在加入L-单甲基精氨酸后,随着PDGF-BB的浓度递增,培养上清中TRAP活性从1.46U/L±0.10U/L升高至最高为2.47U/L±0.38U/L（P<0.01）;骨吸收陷窝的面积和数目分别从436μ     

牙周炎动物模型的研究

目的 :建立一种典型、稳定的牙周炎动物模型。方法 :将 2 4只健康大耳白兔随机分为 4组 ,磨牙颈部结扎钢丝 ,在兔的牙周组织注射 IL - 1-β、TNF-α,以生理盐水为对照 ,分不同时期取材做组织学观察和评价。结果 :实验组 15 d后牙龈组织血管扩张充血 ,牙槽骨改建活跃 ,有大量的骨吸收陷窝及破骨细胞 ,并有新骨形成和成骨细胞出现 ,30 d后仅表现为大量的骨吸收陷窝及破骨细胞 ,无成骨现象 ,且以联合应用 IL - 1-β、TNF-α组最为显著。结论 :IL - 1-β、TNF-α在建立牙周炎动物模型中有重要作用。     

小檗碱对大鼠骨髓源性破骨细胞的分化及骨吸收功能的影响

目的：研究小檗碱对破骨细胞分化及骨吸收功能的影响,探讨小檗碱抑制骨吸收的细胞学基础。 方法：采用原代培养的成骨细胞和骨髓单核细胞联合培养的方法,在1,25-（OH）2维生素D3和地塞米松作用下,使骨髓单核细胞分化形成破骨细胞。通过相差显微镜观察细胞形态,通过抗酒石酸酸性磷酸酶（tartrate—resistant acid phosphatase,TRAP）染色和观察骨片上骨吸收陷窝的形成鉴定破骨细胞。磷酸苯二钠法测定破骨细胞抗酒石酸酸性磷酸酶的活性,计算机图像分析技术测定骨片上破骨性骨吸收陷窝的面积。 结果：小檗碱在0．1～10μmol／L范围内,浓度依赖性地抑制TRAP阳性多核破骨细胞的形成和TRAP活性,减少破骨性骨吸收陷窝的面积;在10μmol／L浓度下,对破骨细胞的抑制作用最强,对TRAP阳性多核破骨细胞的形成和TRAP活性的抑制率分别达到了60．45％和42．12％,骨吸收陷窝面积减少72．69％。 结论：小檗碱通过抑制破骨细胞的形成、分化和骨吸收功能减少骨质的丢失。     

低浓度唑来膦酸对成骨细胞和破骨细胞影响的体外实验

目的：观察低浓度(10-6 mol/L)唑来膦酸(zoledronate acid,ZA)对体外大鼠破骨细胞及成骨细胞的影响。方法体外分别培养大鼠来源的成骨细胞和破骨细胞,将两种细胞各分为两组：空白对照组及低浓度(10-6 mol/L)ZA组。应用抗酒石酸酸性磷酸酶染色、图像分析计算骨吸收陷窝面积,检测破骨细胞形态及骨吸收情况。碱性磷酸酶(alkaline phosphatase, ALP)染色、四甲基偶氮唑盐比色法了解成骨细胞的形态及增殖情况。结果培养1周后破骨细胞具有典型的形态特征,并在骨片上形成了吸收陷窝;ZA组与对照组相比,破骨细胞数量及生成吸收陷窝的数目和面积减少,差异有统计学意义(P＜0.05)。成骨细胞有典型的梭形、ALP染色阳性特征,培养至第7天ZA组成骨细胞光吸收值(3.37±0.11)高于对照组(2.87±0.12),差异有统计学意义(P＜0.05)。结论低浓度(10-6 mol/L)的ZA能够抑制破骨细胞的增殖和活性,促进成骨细胞的增殖,选择恰当给药方式和剂量能够在抑制破骨的同时促进成骨。     

低氧/低氧诱导因子-1α通路对成骨细胞与破骨细胞耦联的调控作用

目的 探讨低氧/低氧诱导因子(HIF) -1α通路对成骨细胞与破骨细胞耦联的调控作用.方法 取出生2～3d条件性基因敲除小鼠颅盖骨的成骨细胞与4～8周龄C57BL/6小鼠股骨破骨细胞的前体细胞,建立基因敲除小鼠成骨细胞与破骨细胞前体细胞共培养体系(野生型、HIF-1α-/-、Vhl-/-、HIF - 1α-/-/Vhl -/-共培养).采用RT-PCR技术检测成骨细胞中核因子κB受体活化因子配体(RANKL) mRNA和骨保护素(OPG) mRNA的表达以及破骨细胞中标志酶基因TRAP mRNA的表达,甲苯胺蓝染色观察破骨细胞溶骨形成的骨陷窝.结果 RT-PCR检测结果显示:与野生型共培养比较,HIF-1α-/-共培养的成骨细胞中RANKL mRNA表达上调、OPG mRNA表达下调(均P＜0.05),破骨细胞TRAP mRNA表达上调;Vhl-/-共培养和HIF-1α-/-/Vhl-/-共培养的成骨细胞中RANKL mRNA表达下调、OPG mRNA表达显著上调(均P＜0.05),破骨细胞TRAP mRNA表达下调;随着共培养时间的延长,成骨细胞中RANKL mRNA和OPG mRNA表达均逐渐减少,而破骨细胞TRAP mRNA表达均逐渐增加.甲苯胺蓝染色倒置显微镜观察显示:共培养第9天,破骨细胞骨吸收陷窝出现;随着共培养时间的延长,骨吸收陷窝面积和深度逐渐增加;与野生型共培养比较,HIF-1α-/-共培养的破骨细胞骨吸收陷窝较大且较深,Vhl-/-和HIF-1α-/-/Vhl-/-共培养的破骨细胞骨吸收陷窝较小且较浅.结论 低氧/HIF-1α通路激活后,成骨细胞抑制破骨细胞的分化功能;低氧/HIF-1α通路阻断后,成骨细胞促进破骨细胞的分化功能.     

葛根素对体外破骨细胞性骨吸收的影响

目的研究葛根素对体外破骨细胞性骨吸收的影响.方法分离破骨细胞,与牛骨磨片共培养,分别加入10-8、10-7、10-6 mol/L的葛根素,以17β-雌二醇10-8 mol/L为阳性对照药,利用相差倒置显微镜观察破骨细胞吸收牛骨磨片的情况.于培养3、5、7 d计数各组骨吸收陷窝数目的变化,并进行显微摄片和图像分析.结果与正常对照组相比,葛根素10-8、10-7、10-6 mol/L及17β-雌二醇10-8 mol/L能使体外培养破骨细胞性骨吸收数目和吸收陷窝面积呈剂量依赖性减少.结论葛根素在体外能直接抑制破骨细胞性骨吸收.     

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.     

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.     

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.