氨基酸聚合物/硫酸钙生物工程骨的生物相容性研究

目的 评价氨基酸聚合物[poly(amino acid),PAA]/硫酸钙(calcium sulfate,CaSO4)生物工程骨的生物相容性.方法 制备PAA/CaSO4.通过急性毒性实验、溶血实验、微核实验、热原实验、皮内刺激实验、细胞毒性实验研究该复合材料的生物相容性.结果 急性毒性实验中各组小鼠活动正常,7 d内小鼠无死亡,各组动物未见中毒症状或不良反应,7 d 后各组小鼠体质量日平均增加差异无统计学意义(P>0.05).材料浸提液的溶血率为2.01%,符合<5%的国家标准.微核实验中3个实验组与阴性对照组微核率无明显差异(各组的微核率均介于0.4～0.6之间),材料无潜在致突变性.热原实验中3只动物中体温升高最高为0.4℃,体温升高的总数为1.1℃,符合热源实验国家标准.皮内刺激实验观察显示,注射材料生理盐水浸提液和生理盐水处无明显红斑、水肿和皮肤坏死,PII 为0.01～0.02,属极轻微刺激.细胞毒性实验中A、B、C组的BMSC的相对增殖率均>90%,细胞毒性为0～1级,材料无细胞毒性.结论 PAA/CaSO4具有良好的生物相容性.     

明胶/壳聚糖创伤敷料的生物安全性评价

目的:对明胶/壳聚糖敷料进行生物安全性评价.方法:进行皮肤刺激,皮肤致敏,细胞毒性和溶血实验,并按照统一的标准判定.结果:皮内注射实验:各组各点局部无红斑、水肿反应;皮肤致敏实验:试验组及空白对照组溶液家兔的致敏率为0%,而阳性对照组为100%,二者相比有显著性差异;溶血实验:除阳性对照组外,各组溶血率均在1%以内,符合规定不大于5%的标准;微核实验:材料浸提液组的微核率为1.6%o,与生理盐水对照组比较无显著性差异(P＞0.05),材料浸提液组与阳性对照组(微核率为20%o)比较有显著性差异(P＜0.01);细胞毒性实验:镜下见纤维细胞生长良好,形态和数量均和对照组无显著性差异,MTT法测得细胞活性和对照组无差别,细胞毒性总评级为0.结论:明胶/壳聚糖生物膜不含致热原、致敏原,无皮肤刺激反应,不引起皮肤过敏反应,无溶血反应,生物安全性保证其可望用于临床.     

新型复合型生物多孔支架生物安全性初步评价

目的 对自制的新型复合型生物多孔支架进行体内、体外生物安全性实验研究,为该材料临床应用的安全性提供理论依据.方法 以自制的新型复合型生物多孔支架为实验组进行以下实验:①采用培养的L929系小鼠成纤维细胞株,以聚乙烯浸提液为阴性对照组,0.64%苯酚溶液为阳性对照组采用细胞观察法与MMT法进行细胞毒性实验.②30只小鼠随机分为3组,以生理盐水为阴性对照组,聚氯乙烯作为阳性对照组进行全身急性毒性实验.③聚乙烯作为阴性对照组,聚氯乙烯为阳性对照组,分别加入抗凝兔血后计算溶血率.结果 细胞毒性试验表明新型复合型生物多孔支架组细胞无明显细胞毒性.全身急性毒性实验实验组和阴性对照组的各指标数值差异无统计学意义,与阳性对照组差异有统计学意义.支架材料的溶血率为3.199%.结论 课题组自制的新型复合型生物多孔支架具有良好的生物相容性,是一种比较好的骨组织产品.     

纳米羟基磷灰石/聚氨基酸复合材料的生物相容性研究

目的:评价新型骨内固定生物材料纳米羟基磷灰石(Nanohydroxy apatite,nHA)/聚氨基酸的生物相容性.方法:对nHA/聚氨基酸复合材料生物相容性进行体外及体内评价,分别进行以下实验:①细胞毒性试验,材料与兔骨髓间充质干细胞共同培养,倒置相差显微镜观察细胞生长,CCK-8检测细胞增殖情况.②急性毒性试验,KM小鼠20只,雌雄不限,体重20～23 g,分两组(n=10),分别按50 g/kg注射材料浸提液及生理盐水,观察小鼠毒性表现.③溶血试验,分3组,分别取10 ml材料浸提液、灭菌蒸馏水、生理盐水,各加入0.2ml新鲜抗凝兔血测溶血率.④热源试验,新西兰大白兔3只,雌雄不限,体重2.3～2.5 kg,按10ml/kg注射材料浸提液,测量正常体温与注射后连续3 h内每小时体温的差值.⑤微核试验,KM小鼠30只,雌雄不限,体重20～23 g,分3组(n=10),分别按100 g/kg间隔24h注射材料浸提液、环磷酰胺及生理盐水2次,观察骨髓嗜多染红细胞及含微核小体的骨髓嗜多染红细胞,计算微核率.结果:nHA/聚氨基酸复合材料对骨髓间充质干细胞无毒性,无急性毒性,溶血率正常,无致热源,无微核试验遗传毒性.结论:nHA/聚氨基酸复合材料具有良好的生物相容性,有望用作骨内固定材料.     

基于MEMS技术的压力感应内藏式膀胱造瘘器的设计与安全性检测

目的:研制一种基于微电子机械系统(Micro electro-mechanical system, MEMS)技术设计,能实时监测膀胱压力又安全的新型内藏式的膀胱造瘘器。方法:运用计算机辅助下MEMS技术设计膀胱造瘘器;按照GB/T16886国家医疗机械生物学评价标准进行细胞毒性实验及致敏实验,评价其安全性。结果:细胞毒性实验显示,实验组细胞毒性反应级别均≤1级,细胞溶解率均<10%,与阴性对照组无明显差别;皮肤刺激实验显示,实验组中,生理盐水浸提液和橄榄油浸提液的原发性刺激指数分别为0.375±0.440与0.375±0.209,对皮肤刺激程度均为极轻微,与空白对照组无明显差别(P分别为0.568与0.765)。结论:该膀胱造瘘器在实验条件下无细胞毒性及致敏性,可进行临床试用,进一步评价其安全性及临床实用性。     

羊脱细胞真皮基质的体内外毒性实验

目的：利用溶血反应、热原反应和细胞毒性反应实验了解羊脱细胞真皮基质(acellular dermal matrix,ADM)在动物体内外的毒性反应强度。方法：溶血反应: 分别制作交联型和非交联型羊ADM的浸提液,为交联组和非交联组; 并设置阳性对照组10 mL灭菌注射用水,阴性对照组10 mL 0.9%氯化钠溶液。 将新鲜抗凝稀释兔血加入实验组和对照组试管中,离心后取上清液测量吸光度值,计算溶血度。热原实验：按照条件入选16只新西兰兔,分为初试A,B两组各3只和复试C,D两组各5只。分别将交联与未交联的两种ADM浸提液注射入两组新西兰兔体内,注射后每隔0.5 h测量一次体温,共测量6次,以6次中最高一次减去正常体温即为该兔的体温升高度数。据此判断是否符合生物制品热原实验的评价标准。MTT实验：收集对数期的小鼠成纤维细胞细胞,种植于含有不同浓度羊ADM浸提液的培养基,测量吸光度值进行细胞活力测定。结果：实验A,B组溶血度均小于5%,热原反应测得新西兰兔体温升高之和低于1.8 ℃。MTT实验显示10%~90%羊ADM浸提液培养基对体外细胞的毒性为1级,100%的羊浸提液对体外细胞的毒性为2级,交联型与非交联型组的浸提液对体外细胞的毒性的无显著性差异(P>0.05)。均对体外细胞的增殖影响轻微。结论：羊ADM植入新西兰兔体内引起的溶血反应和热原反应在评价标准之内,在体外对细胞的增殖和活力影响不明显。     

聚氨酯-BaFe12O19复合微粒血管内栓塞材料的生物相容性

目的 探讨包裹聚氨酯的聚氨酯-BaFe12O19复合型微粒栓塞材料的生物相容性.方法 对聚氨酯-BaFe12O,19复合微粒材料进行一系列生物相容性实验,包括Ames致突变实验、细胞毒性实验、全身急性和亚急性毒性实验、溶血实验、出血和凝血时间测定、凝血功能实验等.结果 Ames实验:该颗粒浸提液各组的MR值均小于2,无致突变性;细胞毒性实验:不同浓度浸提液对L929细胞的细胞毒性均为Ⅰ级;全身急性、亚急性毒性实验:动物一般情况良好,未见异常病理表现;溶血实验中实验组的溶血率为2.43%,符合ISO规定不大于5%的标准;出、凝血时间在实验组与对照组无显著性差异:实验组与对照组的凝血功能未见显著差异(P>0.05).结论 聚氨酯-BaFez1219复合微粒材料无致突变性,无毒性,对出血和凝血时间无显著性影响,不引起溶血和凝血功能的改变.聚氨酯-BaFe12O19微粒具有良好的生物相容性.     

纳米羟基磷灰石/聚酰胺66与人胚胎成纤维细胞的生物相容性研究

目的:研究新型骨修复重建复合材料纳米羟基磷灰石/聚酰胺66(n-HA/PA66)对人胚胎成纤维细胞可能存在的细胞毒性,为该材料应用于临床提供实验依据.方法:采用人胚胎成纤维细胞,经材料及材料浸提液分别与细胞共培养等方式对n-HA/PA66进行细胞毒性测试,细胞形态观察法观察各组细胞在24h、48h、72h、5天各时相点的细胞形态学变化,细胞生长抑制法(MTT比色法),测定各组1,2,4,7天细胞的相对增殖率.结果:材料表面的细胞黏附生长良好,各浸提液组和直接接触组分别与阴性对照组进行两两比较,各组对细胞的增殖均无显著影响(P＞0.05),各浸提液组和直接接触组分别与阳性对照组进行两两比较显示有显著差异(P＜0.05),材料的细胞毒性为0～1级.结论:体外试验结果显示,n-HA/PA66与人胚胎成纤维细胞相容性好,符合GB/T16886.5-1997-ISO 10993-5:1992规定的医用材料相应标准[1].     

核壳型n-HA/ZrO2支架材料对小鼠骨髓PCE微核率的影响

目的探讨新制备的核壳型纳米羟基磷灰石包覆二氧化锆复合生物陶瓷材料(核壳型n-HA/ZrO2支架材料)对小鼠遗传毒性的影响。方法进行微核实验,按50 mg/kg的剂量,从小鼠腹腔注入高、中、低三种剂量核壳型n-HA/ZrO2支架材料浸提液,观察其股骨骨髓每1000个嗜多染红细胞(PCE)中的微核细胞数,并与阴性对照组和阳性对照组进行比较。结果受试样品三种浓度浸提液小鼠骨髓PCE微核率分别为2.2‰(高剂量组),1.8‰(中剂量组)与1.4‰(低剂量组)。统计学分析表明,实验组间PCE微核率比较差异无统计学意义(P>0.05),与阳性对照组比较差异有统计学意义(P<0.001),与阴性对照组比较差异无统计学意义(P>0.05)。结论核壳型n-HA/ZrO2支架材料无致染色体突变及潜在的遗传毒性。     

鸡羽根角蛋白管毒性作用的研究

目的 观察鸡羽根角蛋白管作为组织工程材料的毒性作用.方法 分别制备不同处理强度的3种鸡羽根角蛋白管的浸提液并测定其化学成分.采用皮肤致敏实验(大鼠)、皮内刺激实验(家兔)、小鼠全身急性毒性实验以及体外细胞毒性实验观察鸡羽根角蛋白管浸提液的体内和体外毒性作用.结果 鸡羽根在浸出液中释放的主要物质为中等分子量的肽类物质,大分子蛋白释放不多.皮肤致敏实验和皮内刺激实验显示对浸提液不引起皮肤及皮内红斑、水肿和坏死现象.小鼠全身急性毒性实验中,腹腔注射浸提液的小鼠未出现死亡、衰竭、发绀、震颤、严重的腹膜刺激、眼睑下垂及呼吸困难等毒性症状.体外细胞毒性实验示浸提液细胞毒性接近于0级.结论 鸡羽根角蛋白管可被水解,释放一些肽类物质,其浸提液对皮肤无致敏作用,皮内刺激反应弱,不引起全身急性毒性症状,无细胞毒性.     

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.