大块骨关节软骨冷冻保存的实验研究

目的：探讨大块同种异体骨关节软骨冷冻保存的最佳方法。方法：将高月龄胎儿和成人肢体骨关节移植物用冷冻保护剂预处理后进行分组保存（A组：梯度降温加液氮保存；B组：梯度降温加－80℃低温冰箱保存；C组：单纯液氮保存），定期取部分关节软骨组织进行组织学、组织化学、细胞存活率以及超微结构。结果：A、B组经过冷冻保存1、3、6、12周后关节软骨细胞形态基本正常，软骨基质粘多糖含量丰富，细胞超微结构大体正常，细胞存活率较高；C组经过冷冻后细胞大多出现胞膜破裂、胞核固缩等现象,基质中粘多糖含量降低，细胞器结构不清，细胞存活率低。在A、B组中，胎儿关节软骨中细胞成分较成人多，细胞较幼稚，粘多糖含量更丰富。结论：①梯度降温有利于保持冷冻骨关节移植物中软骨细胞的活性。②胎儿软骨细胞较成人具有强的抗冻伤能力。③软骨细胞的活性与保存时间无关。④液氮和－80℃低温保存对维持软骨细胞活性无明显区别。     

冷冻保存对同种异体移植物免疫原性的影响

同种异体移植物的免疫原性强弱是关系到移植效果的重要因素之一。自从1965年Smith成功地冷冻保存了游离的软骨细胞以及1972年Wilmut等成功地将胚胎长时间冷冻保存以来；一些学者致力于通过冷冻处理改变异体组织的免疫原性，减轻或延缓移植后排斥反应的发生，从而提高移植效果。冷冻又包括低温（高于-80℃）、深低温（-80℃）、超深低温（-196℃）以及反复冻融等方法。现将冷冻对同种异体移植物免疫原性的影响综述如下。     

深低温骨库建立及同种异体骨与关节移植的临床应用

冷冻保存同种异体骨和骨关节在修补大块骨缺损及关节移植的应用中具有重要意义 .采用深低温冷冻法和辐照灭菌法保存的异体骨 .临床应用异体骨时 ,根据患者的需要和骨库的记录资料合理选择 ,临床应用同种异体骨和骨软骨移植手术 4例 ,未见明显排斥反应 ,疗效满意 .     

冷冻同种异体骨软骨移植修复大面积关节软骨缺损的可行性研究

目的观察冷冻同种异体骨软骨移植修复大面积关节软骨缺损的可行性，为临床应用提供实验依据。方法共选取36只新西兰兔，随机数字表法分成供体组、实验组和对照组，每组12只。供体组制备冷冻同种异体胫骨内侧髁骨软骨移植物后处死；实验组将经程序深低温冷冻保存的同种异体胫骨内侧髁移植；对照组将取下的胫骨内侧髁原位植入。所有实验动物在术后12周处死。应用苏木精-伊红染色观察关节软骨组织的病理学变化，软骨缺损的组织学评分以半定量的改良Wakitani score法评估，内容包括：细胞种类、髓质染色、表面完整性、软骨的厚度、移植软骨在受体周围组织的完整性5个方面，采用0、1、2、3、4评分，最高总分14分。观察两组动物的大体标本、X线片、组织学检查、Waki-tani score评分及免疫组织化学检查结果。结果①移植软骨的大体观察：两组动物关节活动度正常，关节腔无粘连、无积液，实验组移植软骨颜色及高度与周围关节软骨大致相同，关节面较平整，基底部界限模糊。对照组关节软骨面平整、无塌陷，厚度不变。②两组动物关节X线片均见骨折线已模糊，对位、对线良好。③组织学检查：两组动物关节软骨均已被透明软骨覆盖，细胞排列有序，软骨基质大量分泌，未见淋巴细胞和浆细胞浸润。④两组动物关节软骨Wakitani score评分结果无显著性差异，总评分相近（P〉0.05）。⑤两组修复软骨Ⅱ型胶原免疫组织化学染色强阳性。结论冻存同种异体骨软骨移植后可完全修复大面积关节软骨缺损，并与自体骨软骨块修复兔的关节软骨缺损在组织学上相近，未见免疫排斥反应。     

深低温冷冻技术对带血管同种异体骨移植作用的实验研究

目的 探讨深低温冷冻技术对带血管同种异体骨移植物抗原性的影响，观察移植骨连接情况。方法 健康成年新西兰白兔36只（随机取12只作供体，24只作受体）分3组：A组（受体12只）深低温冷存带肱血管异体桡骨段移植；B组（受体12只）新鲜带肱血管异体桡骨段移植；C组（供体12只）取移植骨段后继续喂养4周，行免疫学检测，为无移植对照组。术后观察吻合血管通畅率，7，28d检测血清干扰素γ（IFN-γ）水平，2，4，8，12周X线检测，12周行移植骨组织学检查。结果A组血管通畅率、移植骨生长速度和骨连接情况优于B组，血清IFN-γ水平低于B组，骨皮质浅层及深层骨陷窝内骨细胞存活，炎症细胞浸润不明显，血管管腔通畅，B组移植骨未见存活骨细胞，淋巴细胞、浆细胞浸润明显，血管管腔血栓形成。结论深低温冷冻技术降低带血管同种异体骨移植物的抗原性，移植后显著降低器官的移植排斥反应，有助于吻合血管通畅、移植骨的存活，加快骨谇接。     

同种异体软骨细胞复合PLGA支架修复关节软骨缺损的免疫学观察

目的应用同种异体软骨细胞移植修复兔膝关节软骨缺损,评估术后免疫学表现及其修复效果。方法供体为1月龄新西兰兔1只,取其膝关节全层软骨片,0.25%胰蛋白酶和0.2%Ⅱ型胶原酶消化,分离培养软骨细胞。受体为4月龄新西兰兔30只,暴露股骨髁前面关节面,用钻在股骨内髁制作直径4mm、深2mm（达髓腔）的关节软骨缺损模型。软骨细胞移植组植入体外培养12d的PLGA软骨细胞复合体,PLGA移植组只植入PLGA,假手术组只做皮肤切开手术。术后不固定患肢,兔笼中活动。术前、术后3、5、7、12周分离受体外周血淋巴细胞,检测其与异体软骨细胞和PLGA混合后的刺激指数（stimulation index,SI）;术后5、7、12周取修复区软骨及软骨下骨观察局部组织学反应。结果软骨细胞移植组术后检测到淋巴细胞增殖,移植物局部见到淋巴细胞浸润。结论同种异体软骨细胞复合PLGA移植存在免疫反应。     

异种骨软骨移植免疫反应及愈合特点的实验研究

目的了解异种骨软骨移植免疫反应及愈合的特点。方法取成年新西兰白兔36只作为实验动物以及自体骨软骨移植和同种异体骨软骨移植的供体,健康家猪1只作为异种骨软骨移植的供体。将实验动物随机分为A、B、C三组,每组12只,分别为自体组、同种异体组和异种组,在实验动物右股骨滑车部制造骨软骨缺损模型,分别进行柱状自体、同种异体和异种骨软骨移植。分别于术后2周、4周、12周取外周血进行IL-2、IL-4和IFN-γ检测。分别于术后4周及12周随机处死每组中的6只实验动物,取其股骨远端骨软骨、滑膜组织以及关节液标本,进行大体观察、组织学检查和IL-2、IL-4和IFN-γ检测。结果 Moran大体观察评分结果显示,C组在4周时评分低于A组及B组;在术后12周时低于A组,但与B组的评分差异无显著性。血清及关节液中细胞因子检测结果显示,术后2周时,C组血清的IFN-γ浓度低于A组及B组;C组血清的IL-4浓度高于A组及B组。术后4周时,C组关节液中的IFN-γ浓度高于A组;C组关节液中的IL-2浓度高于A组及B组;C组血清的IL-4浓度高于B组,关节液中的IL-4浓度高于A组,但低于B组。术后12周时,C组血清的IFN-γ浓度高于A组及B组,关节液中的IFN-γ浓度高于A组;C组血清的IL-2浓度高于A组及B组,关节液中的IL-2浓度高于A组及B组;C组关节液中的IL-4浓度高于A组。组织学检查结果显示,术后4周时,B组术侧膝关节滑膜组织中有炎细胞浸润,至术后12周时组织学表现基本正常。C组在术后4周时术侧膝关节滑膜组织中有大量炎细胞浸润,12周时炎细胞数量减少,但仍然明显多于B组。同时在A组中也发现个别滑膜标本中存在少量炎细胞浸润。结论异种骨软骨移植的免疫反应特点与同种异体骨软骨移植不同,其早期排斥反应强,后期弱,早期以Th2反应为主,而同种异体骨软骨移植免疫排斥以Th1反应为主。愈合过程则是骨吸收发生较晚,而同种异体移植骨吸收发生较早。     

深低温骨库建立及同种体骨与关系移植的临床应用

冷冻保同种异体骨和骨关节在修补大块骨缺损及关节移植的应用中具有重要意义,采用深低温冷冻法和辐照灭菌法保存的异体骨,临床应用异体骨时,根据患者的需要和骨库的记录资料合理选择,临床应用同种异体骨和骨软骨移植手术4例,未见明显排斥反应,疗效满意。     

深低温处理同种异体肌腱移植组织形态学变化实验研究

目的探讨经深低温冷冻处理(-80℃)、未经处理同种异体肌腱和自体肌腱移植后组织形态学的变化。方法采用兔跟腱缺损修复模型,分为深低温冷冻同种异体肌腱移植组(深低温异体组)、未经处理同种异体肌腱移植组(未处理异体组)和自体肌腱移植组(自体组)。在移植后3周对肌腱进行组织形态学观察。结果光镜、电镜下未处理异体组肌腱腱束变性、坏死,而深低温异体组与自体组则无明显差别。结论深低温冷冻同种异体肌腱移植在形态学方面的表现与自体肌腱移植结果基本相同。     

冻存复苏培养的关节软骨细胞生物学特性

关节软骨（细胞）的同种异体移植是解决关节软骨损伤后修复问题的理想方法．其成功的先决条件要求细胞能够耐受低温冷冻保存并保持关节软骨细胞的生物学特性。为此，该实验在成功地建立了关节软骨细胞库的基础上，进行冻存后的关节软骨细胞培养，同时利用显微缩时录像技术，借助组织学、组织化学和分子生物学等方法．观察细胞的增殖、糖胶聚糖的合成以及Ⅱ型胶原的基因表达情况。结果表明，分离的关节软骨细胞经深低温冷冻保存后复苏．细胞在14d的培养中仍可保持关节软骨细胞所特有的形态以及对糖胺聚糖和Ⅱ型胶原的合成能力。     

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.