纤维环穿刺法诱导兔椎间盘退变模型的组织学和影像学分析

目的探讨纤维环穿刺法诱导椎间盘退变模型的可行性和科学性。方法新西兰大白兔18只,经右侧腹膜外入路用弯血管钳夹持16G穿刺针准确刺入L3-4、L4-5、L5-6椎间盘的纤维环,深度控制在5 mm。于术前及术后4、8、12周对造模后的椎间盘及对照组的椎间盘（以L1-2、L2-3、L6-7为对照）行计算机X线摄影（computed radiography,CR）、MRI检查,并进行组织学观察。结果从手术后第4～12周,造模后的椎间盘高度指数（disc height index,DHI）呈递减趋势,MRI T2WI信号呈现持续减弱趋势（P〈0.01）;组织学观察发现4、8周造模组髓核细胞逐渐减少,12周组髓核细胞几乎被纤维软骨组织替代,纤维环排列不整齐,与髓核界限不清。结论纤维环穿刺法可以诱导兔椎间盘的缓慢退变,为临床深入研究椎间盘的退行性变提供了切实可行的动物模型。     

穿刺抽吸法诱导兔椎间盘退变模型

目的:实验通过纤维环穿刺抽吸法建立兔椎间盘退变模型,并通过病理组织学、影像学检测分析此种方法的特点。方法:取10月龄健康日本大耳白12只,雌雄不限,利用21 G穿刺针分别在兔L3～4、L4～5、L5～6节段刺入纤维环并抽吸髓核约8～10 mg;L1～2、L2～3及L6～7节段只暴露不做穿刺抽吸。造模完毕后,实验兔分别于术后4、8、12、16周各取3只进行指标检测MRI检查,然后处死实验动物获取椎间盘,利用病理组织学观察。结果:L3～4、L4～5、L5～6椎间盘髓核MRIT2WI信号强度在术后随时间延长呈现逐渐降低趋势,髓核面积逐渐缩小,椎间隙高度也逐步下降,从术后4周开始两组差异有统计学意义(P<0.05)。随着时间的进展,番红"O"染色显示纤维环穿刺组髓核内细胞外基质含量逐渐减少。结论:纤维环穿刺法可成功建立椎间盘退变模型,比较真实地模拟了人类椎间盘损伤后的退变过程。     

大鼠尾椎间盘针刺退变模型的建立

目的 建立大鼠尾椎间盘针刺退变模型并进行X线和组织学观察.方法 Wistar 大鼠36只, 使用20G皮肤穿刺针对Co7,8 Co8,9椎间盘分别随机行腹背横贯或半横贯穿刺一针, Co9,10椎间盘不穿刺作为对照, 术前及术后1、2、4周时对造模后的椎间盘及对照的椎间盘(Co9,10)行X线检查及HE染色观察.结果 术后第1周到第4周,穿刺后的椎间盘高度指数呈现持续减小; 组织学检查发现横贯穿刺和半横贯穿刺椎间盘1周时都发生退变,而横贯穿刺退变较严重(P＜0.01).结论 横贯或半横贯针刺方法可以成功建立大鼠尾椎间盘退变模型,方法可靠、有效、重复性好.     

椎间盘内注射碱性成纤维细胞生长因子对大鼠腰椎间盘针刺退变的预防作用

目的: 观察碱性成纤维细胞生长因子（basic fibroblast growth factor, bFGF）对SD大鼠腰椎间盘针刺退变的预防效果。方法： 24只成年SD大鼠随机分为实验组、对照组、空白组,每组8只。实验组在术中透视引导下用27G针头对L4-5,L5-6椎间盘后侧针刺后以微量注射器向椎间盘内注射bFGF 50 μL, 对照组椎间盘穿刺后注射生理盐水50 μL,空白组椎间盘不予穿刺,术后8周3组SD大鼠统一行MRI检查,观察T2加权像腰椎间盘信号强度的改变,以Pfirrmann分级法对椎间盘退变程度进行分级,取出椎间盘行病理学观察。结果： MRI结果显示,术后8周实验组及空白组髓核信号强度明显高于对照组,各组间Pfirrmann评分结果差异有统计学意义（P<0.05）,病理学检查显示实验组及空白组髓核细胞数量无明显改变,纤维排列整齐,对照组髓核细胞数量明显降低,胶原纤维排列紊乱。结论：bFGF注射对于后侧纤维环针刺术后SD大鼠椎间盘的退变有预防作用。     

经皮微创纤维环穿刺法制作兔椎间盘退变模型

目的应用经皮微创穿刺方法制造兔腰椎间盘退变模型。方法在C型臂X光机监控下,定位18只兔子的L3/4、L4/5、L5/6椎间盘,应用经皮穿刺方法穿刺纤维环,每组行MRI检查后予以处死,取L3/4、L4/5、L5/6椎间盘髓核组织行HE染色观察髓核细胞,用免疫组化染色法观察Ⅱ型胶原蛋白染色情况,取L2/3及L6/7正常间盘组织作为对照。结果造模前后椎间盘髓核的MRI信号强度有随造模手术后的时间延长而逐渐降低的趋势(L3/4、L4/5及L5/6,P<0.05)。病理HE染色观察发现,对照组正常的髓核组织中有大量的髓核细胞,随着通过微创法制作兔腰椎间盘退变动物模型后时间的延长纤维环细胞、髓核细胞数量逐渐减少,到第12周时,髓核组织中只有很少量的髓核细胞,却有很多纤维软骨组织。对照组及术后4、8、12周组Ⅱ型胶原免疫组化染色的灰度值单因素方差分析结果 F=41.82,P<0.05,组间比较LSD-t检验结果显示除术后4周与8周组比较差异无统计学意义外,其余组间比较均有显著差异(P<0.05)。结论成功建立了新的微创兔腰椎间盘退变动物模型。     

大鼠尾椎间盘穿刺诱导椎间盘退变动物模型的实验研究

目的探讨建立大鼠尾椎间盘穿刺诱导椎间盘退变动物模型的可行性。方法 SD大鼠24只,用20G穿刺针腹背贯通穿刺大鼠7/8尾椎间盘。术后1、2、4周对造模后的椎间盘及对照组的椎间盘行免疫组化及组织学观察。结果术后第1周到第4周,造模组椎间盘组织学及免疫组化观察发现髓核细胞的数量及Ⅱ型胶原含量较对照组进行性减少。结论大鼠尾椎间盘穿刺法可以诱导椎间盘的缓慢退变,为研究椎间盘的退行性变提供有效的动物模型。     

肿瘤坏死因子-α作用下椎间盘退变动物模型的建立

目的:制作家兔椎间盘退变（IVDD）的动物模型,研究肿瘤坏死因子-α（TNF-α）对髓核组织的影响。方法:取健康成年家兔30只,手术暴露L₂~L₆,L_3/4、L_4/5作为实验组,每个椎间盘髓核注射20 ng/μL TNF-α 15 μL;L_5/6作为对照组,每个椎间盘髓核注射15 μL磷酸缓冲盐溶液;L_2/3作为空白组,不做处理。于术后4周、8周、12周行腰椎X线及磁共振（MRI）检查,每次随机检查10只家兔,检查完后处死并取其髓核组织经行免疫组织化学及原位末端标记检查。结果:术后4~12周,X线见实验组椎间隙高度进行性下降,MRI可见实验组椎间隙T2加权像信号逐渐降低;免疫组织化学及TUNEL显示实验组髓核组织内Ⅰ型胶原及细胞凋亡率随之时间推移逐渐增加,Ⅱ型胶原逐渐减少。对照组和空白组差异无统计学意义（P>0.05）。结论:实验成功建立了TNF-α作用下的IVDD动物模型。TNF-α介导的椎间盘术后椎间隙高度丢失,同时TNF-α可诱导椎间盘髓核细胞退变及凋亡。微量注射器的穿刺作用对髓核细胞的退变及凋亡无明显影响。     

纤维环穿刺法建立兔椎间盘退变模型

目的:拟建立一个有效的可重复的椎间盘退变模型.方法:取健康成年新西兰大白兔40只,体质量2.5~3.0 kg,雌雄不限,随机分为纤维环穿刺组和对照组,每组20只.经腹膜外入路暴露L4/5、L5/6两个椎间隙,采用21号针头从椎间隙侧前方刺入L4/5、L5/6椎间盘的纤维环,深度控制在5 mm.对照组仅分离暴露椎间盘,不做任何处理.术后1、2、4、6、8周每组随机选取4只兔子行MRI、HE染色及Ⅱ型胶原免疫组化观察退变椎间盘的变化.结果:MRI观察发现,纤维环穿刺组术后椎间盘T2信号强度呈持续减弱趋势,椎间隙高度也不断下降.从术后2周开始两组差异有统计学意义(P<0.05).HE染色观察发现,随着时间进展,纤维环穿刺组髓核细胞逐渐减少,8周时组髓核组织几乎完全变性,被纤维软骨组织替代.Ⅱ型胶原免疫组化染色示,纤维环穿刺组Ⅱ型胶原表达随时间呈进行性下降,从术后第2周开始两组差异有统计学意义(P<0.05).结论:纤维环穿刺法可以诱导兔椎间盘缓慢退变,为深入研究椎间盘退变机制及治疗手段提供可靠的动物模型.     

肌间隙入路腰椎间盘退变模型的构建

目的探讨背最长肌与腹外斜肌间隙入路纤维环穿刺法构建兔腰椎间盘退变模型的可行性。方法新西兰大白兔6只,从兔子背最长肌与腹外斜肌间隙入路暴露椎体和椎间盘,L2/3为对照组,不暴露也不穿刺椎间盘;L3/4为假手术组,剪断L4右侧横突后暴露椎间盘,但不进行穿刺;L4/5、L5/6为穿刺组,剪断L5、L6右侧横突后暴露椎间盘并用18G针穿刺构建模型。分别在术前及术后4周对实验动物进行X线和磁共振(MRI)检查,4周后处死实验动物,取材后行组织学及免疫组织化学检查。结果术后4周,X线检查显示模型组L4/5、L5/6椎间盘高度明显降低、骨赘形成、终板硬化,MRI T2加权像亦见椎间盘高度下降,其椎间盘髓核信号明显减弱;大体观察见该组椎间盘髓核组织较少并组织松脆,呈淡黄色改变;组织学观察髓核脊索细胞数量减少,免疫组织化学观察到髓核Ⅱ型胶原表达比对照组和假手术组明显减少。结论本研究为椎间盘退变的研究提供了一种较为理想的、相对可靠的动物模型。     

纤维环穿刺法建立兔腰椎间盘退变模型

目的 建立纤维环穿刺法建立兔椎间盘退变模型。方法 40只新西兰大白兔随机分为牵引组和对照组,手术组（30）只以16G针刺损伤新西兰兔腰椎椎体右前侧纤维环制备椎间盘退变模型,对照组（10只）不予穿刺手术。分别于造模术后4、8、12周通过腰椎侧位X片及组织病理学检查观察椎间隙变化情况及组织病理情况。结果 手术组在腰椎侧位X线片上术后4周椎间隙高度逐渐下降,椎间盘高度指数变化率（DHI）2组差异有统计学意义（P<0.05）;且随着时间的推移,手术组椎间盘内细胞逐渐减少。结论 纤维环穿刺法可缓慢模拟人类损伤后椎间盘退变,操作安全可靠。     

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.