雪旺细胞及神经干细胞与Chitosan-PLGA导管的细胞相容性

目的研究雪旺细胞(SCs)、神经干细胞(NSCs)与内置纵行排列纳米纤维丝的静电纺丝壳聚糖-聚羟基乙酸(Chitosan-PLGA)纳米导管材料的细胞相容性,探讨细胞在导管材料上的黏附及定向生长情况。方法实验分为SCs组、NSCs组、SCs-NSCs共培养组,3组细胞分别与Chitosan-PLGA纳米导管材料复合培养,四甲基偶氮唑蓝(MTT)比色法观察各组细胞存活率,扫描电镜下观察SCs、NSCs在Chitosan-PLGA上的生长。结果细胞存活率排序由高至低依次为共培养细胞组、SCs组、NSCs组。SCs及NSCs能贴附在Chitosan-PLGA内置纳米纤维丝上生长并沿纤维丝定向生长。结论雪旺细胞及神经干细胞共培养可促进细胞与Chitosan-PLGA材料的相容性。内置纵行排列的纳米纤维丝有引导细胞定向生长与迁移的作用。     

不同质量比SF/PLLA复合纤维支架上共培养的上皮细胞和成纤维细胞增殖和表型的变化

目的: 观察不同质量配比的丝素蛋白(SF)和聚乳酸(PLLA)构建的SF/PLLA复合纤维支架对上皮细胞和成纤维细胞增殖和表型的影响,阐明SF/PLLA生物工程膜的细胞相容性。方法: 使用SF和PLLA作为支架材料,以SF与PLLA不同质量比(S50/P50、S40/P60和S30/P70)进行混合纺丝,制备出不同比例的SF/PLLA复合纤维支架,采用SF/PLLA复合纤维支架与皮肤组织中分离的原代上皮细胞和成纤维细胞进行共培养。采用CCK8方法检测细胞在不同质量比的支架材料上的增殖活性,免疫荧光染色观察共培养细胞的特异标记蛋白表达。结果: 共培养第5和6天时,在S50/P50组复合纤维支架上的成纤维细胞增殖活性高于S40/P60组和S30/P70组(P<0.01),在S40/P60组复合纤维支架上的上皮细胞增殖活性明显高于S50/P50和S30/P70组(P<0.05或P<0.01)。接种于SF/PLLA复合纤维支架的上皮细胞表达CK19蛋白,接种的成纤维细胞表达Vimentin 蛋白。2种细胞与SF/PLLA复合纤维支架共培养,S40/P60和S50/P50组复合纤维支架上的细胞生长状态优于S30/P70组。结论: 支架中SF的浓度增加可以明显提高细胞增殖活性,促进细胞存活,SF/PLLA复合纤维支架对细胞的生物表型无明显影响。     

聚癸二酸丙三醇酯对聚乳酸的改性

以丙三醇和癸二酸为单体通过熔融缩聚制得了聚癸二酸丙三醇酯(PGS),并用其预聚物(p-PGS)对聚L-丙交酯(PLLA)进行共混改性.利用傅立叶变换红外光谱(FT-IR)、核磁共振氢谱(1H-NMR)及凝胶渗透色谱(GPC)法对P-PGS的结构进行表征,并研究了改性后材料的力学性能、两相相容性、亲水性能和细胞相容性.结果表明:P-PGS具有支化分子结构,分散系数约为2.7;共混改性后的材料弹性模量和拉伸强度均有所下降,而断裂伸长率从7 %显著提高到150%左右;PLLA/PGS属于海岛式共混结构,PGS以小于10μm的尺寸均匀分布在PLLA基体中;共混后材料的亲水性也有一定的提高,且几乎保持了PLLA原有的细胞相容性.     

雪旺细胞与PDLLA/CS/CHS自组装复合材料生物相容性研究

目的利用自行培养的雪旺细胞(SCs)与具有良好可生物降解性的高分子聚乳酸/硫酸软骨素/壳聚糖(PDL-LA/CS/CHS)复合材料进行生物相容性研究,评价该材料应用于周围神经修复的可能性。方法利用双差速贴壁法进行SCs的培养与纯化,并观察其生长曲线,苏木精-伊红染色和免疫组织化学染色观察细胞表征;将纯化后的SCs接种在PDLLA/CS/CHS复合材料上进行材料生物相容性研究,用MTT法和环境扫描电镜检测和观察细胞生长情况。结果 SCs生长平台期为1 d,对数期为5~7 d,倍增时间为5 d,其纯度超过90%。MTT实验显示,细胞接种0、2、4 d,PDLLA组、CHS组和PDLLA/CS/CHS组的吸光度值均低于SCs对照组,至7 d和10 d,PDLLA、CHS组吸光度值亦低于SCs对照组,PDLLA/CS/CHS组高于SCs对照组,但差异无统计学意义(P>0.05),PDLLA/CS/CHS组与PDLLA组相比差异有统计学意义(P<0.05)。结论 PDLLA/CS/CHS复合材料是具有较理想的生物相容性的生物材料,有良好的材料-细胞界面,较利于SCs黏附及生长、增殖,可应用于周围神经修复的研究。     

PLLA纳米纤维担载紫杉醇对大鼠胶质瘤C_6细胞株毒性研究

目的:探讨PLLA纳米纤维担载紫杉醇对体外培养大鼠脑胶质瘤C6细胞株的抑制杀伤作用及机理。方法:采用细胞毒试验(MTT法)、流式细胞术(FCM)及形态学观察,研究紫杉醇对脑胶质瘤C6细胞的毒性作用。结果:PLLA纳米纤维担载紫杉醇作用于脑胶质瘤C6细胞后,细胞增殖活力显著下降。有丝分裂周期被阻滞于G2/M期,DNA含量≥4C或DNA含量<2C的细胞比例增加。肿瘤细胞出现核固缩、核碎裂及多核细胞。结论:PLLA纳米纤维担载紫杉醇对胶质瘤细胞有肯定的抑制杀伤作用,药物对肿瘤细胞的毒性效应表现为时间依赖性与剂量依赖性。     

大鼠原代背根神经元和雪旺细胞在随机及有序排列聚甲基丙烯酸甲酯电纺纳米纤维上共培养的形态学

目的: 探讨聚甲基丙烯酸甲酯(PMMA)电纺纳米纤维支架的拓扑线索对于大鼠原代背根神经元(DRGn)培养及其与雪旺细胞(SCs)共培养的影响。 方法: 构建具有随机分布和轴向有序排列拓扑结构的PMMA电纺纳米纤维,分别为随机和有序PMMA电纺纳米纤维组,以PMMA薄膜作为对照组;分离纯化大鼠原代DRGn和SCs,与上述各组PMMA电纺纳米纤维共培养;利用慢病毒技术转染荧光蛋白基因作为显色方法,观察PMMA电纺纳米纤维的拓扑线索对于DRGn神经突生长的影响,在共培养实验通过荧光图像的快速傅立叶转换(FFT)及半高全宽值(FWHM)的计算,定量分析电纺纤维对DRGn神经突和SCs细胞突起的接触引导作用。 结果: 大鼠原代DRGn和SCs均能够顺利在PMMA材料上贴壁并生长;与PMMA薄膜组比较,随机和有序PMMA电纺纳米纤维组DRGn平均神经突数量及神经突长度差异无统计学意义(P>0.05);共培养实验中,电纺纤维的拓扑线索对于 DRGn神经突和SCs细胞突起的生长均具有明显的接触引导作用,与PMMA薄膜组和随机PMMA电纺纳米纤维组比较,有序PMMA电纺纳米纤维组DRGn和SCs的FWHM值均明显降低 (P<0.01),有序PMMA电纺纳米纤维能够在空间上促成DRGn神经突和SCs细胞突起建立共定位。 结论: 有序PMMA电纺纳米纤维具有作为脊髓损伤(SCI)后植入性支架材料的潜力,其拓扑线索有可能加速SCs的轴突髓鞘化过程。     

PLCL/纤维蛋白原静电纺丝膜片与兔骨髓间充质干细胞的体外生物相容性

目的研究PLCL/纤维蛋白原混纺膜片与兔骨髓间充质干细胞的体外生物相容性。方法通过静电纺丝法制备PLCL/纤维蛋白原纳米纤维膜片,并通过扫描电镜(SEM)及水接触角分析仪进行表征。以种植在PLCL/纤维蛋白原膜片中的细胞作为实验组,以种植在聚苯乙烯培养板中的细胞作为对照组,并通过MTT法检测细胞在两组中的黏附、增殖情况。结果分离培养的细胞符合间充质干细胞特性,PLCL/纤维蛋白原纳米纤维的平均直径为(305±87)nm、水接触角为(70.21±2.13)°,与聚苯乙烯相比,其对细胞的生长无明显影响。结论 PLCL/纤维蛋白原混纺膜片具有较好的生物相容性,在血管组织工程中有较大的应用潜力。     

聚己内酯/明胶复合纳米材料的细胞相容性

足底软组织创伤在临床上比较常见,尽管创伤外科修复重建足底软组织缺损的术式比较成熟,但是对术后功能的恢复仍不满意,出现局部软组织稳定性差,使得患者康复后出现站立不稳等问题。恢复足底软组织特有的功能,尤其是稳定性,是临床难以解决的问题。组织工程改变了传统"以创伤修复创伤"的治疗模式,为最终实现无损伤修复创伤开辟了新途径。由于纳米纤维不仅具有表面积大、孔径小、孔隙率高、较好的均一性、直径与细胞外基质中胶原纤维相近,最大程度地模仿细胞外基质天然结构。因此,实验用聚己内酯(PCL)和明胶(gelatin)为原料,应用静电纺丝法制作PCL/gelatin复合纳米纤维(图1)。这样不仅避免了单用PCL导致降解慢,生物相容性差,还避免了单用明胶导致降解太快的问题。通过调节PCL与gelatin不同的混合比例,从而得到需要的降解率和生物相容性。实验分为材料组和对照组两组,将真皮成纤维细胞分别接种在材料组和对照组上,在48 h、72 h、96 h后用MTT法检测细胞的增殖情况。结果显示,真皮成纤维细胞在PCL/gelatin复合纳米材料表面贴附牢固,生长形态良好。MTT检测结果统计表明此材料能够促进真皮成纤维细胞在其表面增殖(图2)。认为PCL/gelatin复合纳米材料具有较好的生物相容性,有望作为修复足底软组织创伤的支架材料。     

PLGA新型纳米支架的制备及其生物相容性

目的探讨和比较应用不同参数静电纺丝技术制备复合型聚乳酸-羟基乙酸[poly(lactic-co-glycolicacid),PLGA]纳米纤维缓释膜片的可行性,检测其表面特性以及生物相容性。方法使用不同参数制备纳米纤维膜,扫描电镜观察膜片表面形态;人牙周膜细胞(human periodontal ligament cells,hPLDCs)培养及鉴定后接种于空白膜上,扫描电镜观察细胞与膜的附着情况,以MTT[3-(4,5-dimethylthiaozol-2-yl)-2,5-dip henyltetrazolium bromide]实验检测不同浓度材料浸提液对细胞增殖变化情况的影响。结果随着PLGA流量增加,纳米纤维直径增加;细胞与PLGA膜复合后粘附良好且增殖状态无明显变化;不同浓度材料浸提液对牙周膜细胞增殖的影响无明显差异。结论通过不同参数静电纺丝制备的新型纳米纤维膜具有良好的生物相容性,可进一步作为多种药物及细胞因子载体构建缓释型支架,从而用于引导性牙周组织再生(guided tissue regeneration,GTR)或者牙周组织工程。     

In vitro biocompatibility between electrospun nanofibrous films of PLCL/fibrinogen and rabbit bone marrow mesenchymal stem cells

目的 研究PLCL/纤维蛋白原混纺膜片与兔骨髓间充质干细胞的体外生物相容性.方法 通过静电纺丝法制备PLCL/纤维蛋白原纳米纤维膜片,并通过扫描电镜(SEM)及水接触角分析仪进行表征.以种植在PLCL/纤维蛋白原膜片中的细胞作为实验组,以种植在聚苯乙烯培养板中的细胞作为对照组,并通过MTT法检测细胞在两组中的黏附、增殖情况.结果 分离培养的细胞符合间充质干细胞特性,PLCL/纤维蛋白原纳米纤维的平均直径为(305±87) nm、水接触角为(70.21±2.13)°,与聚苯乙烯相比,其对细胞的生长无明显影响.结论 PLCL/纤维蛋白原混纺膜片具有较好的生物相容性,在血管组织工程中有较大的应用潜力.     

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.