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.     

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。     

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。     

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。     

肌间隙入路腰椎间盘退变模型的构建

摘要：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。
     

枸杞多糖对大鼠腰椎间盘退变影响的形态学观察

目的探讨枸杞多糖(LBP)对大鼠退变腰椎间盘的形态学的影响。方法将健康雄性SD大鼠42只,随机分为假手术组、椎间盘退变组、LBP干预组,每组14只。采用手术方法制作大鼠腰椎间盘退变模型,LBP干预组在造模后4周,每天上午8:00灌胃给药,80mg.mL-1,容量为1mL.kg-1,连续4周。假手术组及椎间盘退变组分别给予等容量生理盐水灌胃,连续4周。在造模后8周三组所有大鼠再次麻醉,取出椎间盘组织,分别在光镜及电镜下观察椎间盘形态学的变化。结果 HE染色光镜观察假手术组髓核细胞丰富,形态不规则,集中成簇;椎间盘退变组髓核细胞稀少,细胞基质见大量的空泡;LBP干预组髓核细胞有变性、坏死,但较椎间盘退变组退变程度减轻。电镜下假手术组的髓核细胞多见,核膜完整,细胞器丰富;椎间盘退变组髓核细胞减少,细胞器缺失,见大量的空泡;LBP干预组髓核细胞减少,有变性、坏死,退变较椎间盘退变组减轻。结论 LBP可能对大鼠腰椎间盘髓核组织退变有延缓作用。     

枸杞多糖对实验性退变腰椎间盘中基质金属蛋白酶-3表达的影响

目的探讨枸杞多糖(LBP)对大鼠退变腰椎间盘中基质金属蛋白酶-3(MMP-3)表达的影响。方法将雄性SD大鼠42只,分为假手术组、模型组、LBP药物组,每组14只。采用手术方法制作大鼠腰椎间盘退变模型,LBP药物组在造模后4周,每天上午8时灌胃给药,80 mg/kg,容量为1 ml/kg,连续4周。假手术组及模型组分别灌胃给予等容量生理盐水,连续4周。在造模后8周3组所有大鼠再次麻醉,取出椎间盘组织,HE染色观察椎间盘的形态学变化,免疫组织化学法检测腰椎间盘中MMP-3表达的变化。结果形态学观察结果显示,假手术组髓核细胞数目较多,形态多样性;模型组髓核细胞稀少,细胞基质见大量的空泡;LBP药物组髓核细胞有不同程度的变性、坏死,但较模型组退变程度轻。免疫组织化学结果显示,模型组腰椎间盘组织中MMP-3阳性染色强度高于假手术组(P<0.05);LBP药物组腰椎间盘组织中MMP-3阳性表达较模型组低(P<0.05),而与假手术组比较差异无统计学意义(P>0.05)。结论 LBP抑制大鼠退变腰椎间盘组织中MMP-3的表达,可能对细胞外基质中蛋白多糖有间接降解的作用,从而延缓或抑制实验性椎间盘的退行性改变。     

转基因髓核细胞移植对兔腰椎间盘退变的影响

目的：椎间盘退变是引起腰背痛的主要原因。将腺相关病毒（adeno-associated virus,AAV）介导的结缔组织生长因子（connective tissue growth factor,CTGF）和基质金属蛋白酶组织抑制因子1(tissue inhibitor of metalloproteinases 1,TIMP1) 双基因体外联合转染兔椎间盘髓核细胞后,再移植于退变椎间盘内,观察椎间高度、II型胶原和蛋白多糖合成的变化,以探讨体外延缓或逆转椎间盘细胞退变的方法。方法 20只新西兰大白兔应用CT引导下的微创穿刺法建立兔椎间盘退变模型。以rAAV2-CTGF-IRES-TIMP1体外转染培养的兔髓核细胞,再将其显微注射于退变椎间盘内作为细胞移植组;退变椎间盘内注入磷酸盐缓冲液（PBS）作为退变对照组;正常椎间盘作为空白对照组。分别于6、10、14周行X线和MRI检查观察椎间高度的变化及椎间盘信号的改变,Western-blot鉴定细胞因子CTGF和TIMP1在椎间盘内的表达,35S整合法测定兔髓核组织蛋白多糖合成率,RT-PCR检测II型胶原及蛋白多糖mRNA的表达。结果 MRI证实CT引导下经皮微创穿刺2周后椎间盘开始退变。与退变对照组相比,MRI显示细胞移植组椎间盘退变明显减轻,转基因兔髓核细胞移植可减缓椎间高度(DHI)下降的发生率,转基因细胞移植组较退变对照组相比,Ⅱ型胶原含量和蛋白多糖的生物合成明显增加,差别有统计学意义（P﹤0.05）。结论 经皮CT 引导下穿刺可成功建立兔椎间盘退变模型。转双基因CTGF和TIMP1 髓核细胞移植可有助于椎间高度的维持,促进椎间盘内蛋白多糖和II型胶原的生物合成,明显延缓椎间盘退变。     

兔腰椎间盘退变模型的建立及影像学分析

目的探讨针刺抽吸法建立椎间盘退变动物模型的可行性。方法新西兰大白兔18只,用持针器夹持21 G皮肤穿刺针从椎间隙正前方刺入L3-4、L4-5、L5-6椎间盘的纤维环,深度控制在5 mm,拔出针芯,抽吸出部分髓核组织。术前及术后4、8、12周分别对造模后椎间盘及对照椎间盘（L2-3）行MRI及CR检查。结果术后第4周到第12周,造模后的椎间盘MRI的T2W1信号呈现持续减弱趋势;椎间盘高度指数持续下降,差异有高度统计学意义（P〈0.01）。结论针刺抽吸法可诱导兔椎间盘的缓慢退变,从而为研究椎间盘退行性变的机制及治疗提供有效的动物模型。     

白藜芦醇对大鼠腰椎间盘退变的影响

目的探讨白藜芦醇(RES)对大鼠退变腰椎间盘病理组织学改变及基质金属蛋白酶-9(MMP-9)表达的影响。方法雄性SD大鼠36只,随机分为正常对照组、椎间盘退变组和白藜芦醇药物干预组,每组12只,手术方法制作大鼠腰椎间盘退变模型。白藜芦醇药物干预组在造模后4周,将RES超细粉末制成20mg.mL-1的溶液,按50 mg.kg-1,每天1次腹腔注射,持续2周。正常对照组和椎间盘退变组给予腹腔注射生理盐水,三组大鼠在造模后8周取腰椎间盘组织,HE染色观察各组椎间盘病理组织学改变;免疫组织化学法观察MMP-9的表达。结果 HE染色显示白藜芦醇药物干预组可见不同程度的髓核的脊索细胞及软骨样细胞减少,周围基质和水分亦较少,但比椎间盘退变组退变程度轻。免疫组织化学染色结果 MMP-9阳性染色的灰度值分析显示白藜芦醇药物干预组阳性染色弱于椎间盘退变组(P<0.05),与正常对照组比较差异无统计学意义(P>0.05)。结论白藜芦醇具有减轻实验性腰椎间盘组织退变的作用,抑制MMP-9的表达可能是其作用机制之一。     

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.