VEGF同轴静电纺丝膜的体外血液相容性及生物学活性研究

目的 评价VEGF同轴静电纺丝膜的物理特性、体外血液相容性和生物学活性,探求其作为人工心血管替代材料的可行性.方法 通过静电纺丝技术制备VEGF同轴静电纺丝膜,然后进行体外溶血实验、动态凝血实验、凝血时间测定、血小板粘附实验、细胞毒性实验、体外细胞粘附实验对其血液相容性和生物学活性进行评价,并与涤纶(Dacron)作平行对照.结果 VEGF同轴静电纺丝膜的血液相容性较高,经统计学分析与Dacron之间差异无统计学意义(P＞0.05).VEGF同轴静电纺丝膜的体外细胞粘附实验中,经细胞计数Dacron为(5.34±0.94)×103,PCL-PEG静电纺丝膜为(7.82±0.85)×104,PCL-PEG/BSA同轴静电纺丝膜为(8.04±0.84)×104,PCL-PEG/BSA-VEGF同轴静电纺丝膜则为(1.39±0.76)×105,经统计学分析PCL-PEG静电纺丝膜和PCL-PEG/BSA同轴静电纺丝膜之间差异无统计学意义(P＞0.05),PCL-PEG静电纺丝膜和PCL-PEG/BSA同轴静电纺丝膜与Dacron之间差异有统计学意义(P＜0.05),而PCL-PEG/BSA-VEGF同轴静电纺丝膜与PCL-PEG静电纺丝膜和PCL-PEG/BSA同轴静电纺丝膜之间的差异具有统计学意义(P＜0.05).结论 VEGF同轴静电纺丝膜血液相容性较高,且更易于与内皮细胞粘附,显示其成为人工心血管替代材料的前景.     

牙周引导组织再生壳聚糖膜的生物相容性研究

目的 :观察壳聚糖作为牙周引导组织再生膜的生物相容性。方法 :采用冷冻干燥法制备壳聚糖膜 ,然后通过细胞毒性试验、溶血试验、热原试验、过敏试验等体内外生物学试验相结合的方法研究其生物相容性。结果 :测试表明壳聚糖膜无毒、不溶血、不致热、不致敏。结论 :壳聚糖膜具有良好的生物相容性。     

静电纺丝聚乳酸-乙醇酸聚合物膜与大鼠嗅鞘细胞的生物相容性研究

目的 探索静电纺丝技术制作的聚乳酸-乙醇酸聚合物（PLGA）膜与大鼠嗅鞘细胞（olfactory ensheathing cells,OECs）的生物相容性。方法 体外培养纯化成年大鼠OECs,将细胞接种于静电纺丝PLGA膜上,使用相差显微镜观察静电纺丝PLGA膜上OECs的细胞形态与分布;用CFDA SE荧光染色检测接种后1～5 d内静电纺丝PLGA膜上OECs的增殖情况;以多聚赖氨酸（PLL）包被细胞做对照。将静电纺丝PLGA膜植入大鼠体内观察其与大鼠的组织相容性。结果 体外培养纯化所得的OECs的细胞纯度大于90%。相差显微镜下示OECs在静电纺丝PLGA膜孔隙中的纳米纤维上黏附良好,细胞状态佳,且特异性地沿纳米纤维方向生长。接种后1～5 d内OECs在静电纺丝PLGA膜上均正常增殖,细胞数目与对照组相比差异无统计学意义。静电纺丝PLGA膜局部移植后大鼠无死亡,PLGA膜与周围组织融合并部分降解。结论 静电纺丝PLGA膜具有良好的生物相容性,OECs在其膜上的黏附和增殖状况良好,是有潜力的神经修复组织工程化神经移植物。     

电纺β-磷酸三钙／明胶引导组织再生膜的制备及研究

目的：制备一种β-磷酸三钙（β-TCP）与明胶（Gel）杂化的纳米纤维引导组织再生膜,并对其生物相容性进行研究,为新型牙周组织再生材料的研制提供理论依据．方法：采用静电纺丝技术制备β-磷酸三钙与明胶的杂化纤维膜．通过扫描电子显微镜（SEM）和力学拉伸实验对杂化纤维膜进行表征,通过MTT实验及细胞与材料共培养对其生物相容性进行评价．结果：杂化纤维膜交联前、后其纤维的平均直径分别为200～300nm和400～500nm,在交联之后其拉伸断裂应力从（1．08±0．24）MPa提高到（6．47±0．85）MPa．杂化纤维膜在细胞毒性上与阳性对照间有统计学差异（P〈0．01）,与阴性对照无统计学差异,并且人成骨样细胞（MG-63）可以成功的贴附生长在该材料上．结论：β-磷酸三钙／明胶纳米纤维引导组织再生膜可以成功地用电纺丝法制备出来,且其生物相容性较好,表明该材料可以用于组织工程学的研究,是一个具有很好应用前景的牙周组织工程再生材料．     

小口径聚氨酯人工血管的制备及其力学性能、细胞相容性评价

目的使用静电纺丝法构建一种小口径聚氨酯血管替代物,观察人工血管的微观结构,并检测其力学性能和细胞相容性。方法以聚氨酯为原料,通过静电纺丝法,以高速旋转的转轴为收集装置,制备4 mm口径的PU人工血管。探讨纺丝液质量分数对纤维直径的影响,滚轴转速对纤维排列的影响,以及质量分数、纤维取向、管壁厚度对人工血管孔隙率的影响,并检测人工血管的力学性能及细胞相容性。结果静电纺丝法制备的PU人工血管具有由多层微纳级超细纤维叠加而成的三维多孔网状结构,平均孔隙率为(51.48±4.47)%,轴向抗拉强度为(5.85±0.62)Mpa,无细胞毒性,并有利于内皮细胞黏附及增殖。结论使用静电纺丝法制备小口径聚氨酯人工血管是可行的,具有潜在的临床应用前景。     

PLGA/鱼皮胶原共轭静电纺丝膜在大鼠体内降解的实验研究*

目的：对一种新型聚乳酸-羟基乙酸(polylactic acid-glycolic acid, PLGA)/鱼皮胶原共轭静电纺丝膜的细胞毒性和体内降解进行初步探讨,为应用于引导骨组织再生(guide bone regeneration, GBR)提供实验依据。方法：以PLGA和医用级鱼皮Ⅰ型胶原为原材料,通过共轭静电纺丝技术制备出高取向性的纳米纤维膜,然后分别检测细胞毒性和大鼠体内的降解过程,初步评价组织学形态与体内降解率。结果：PLGA/鱼皮胶原共轭静电纺丝膜的细胞毒性为1级,符合植入性医用材料的标准。成纤维细胞黏附生长良好,且不能穿过纤维膜生长到膜的对侧。结论：PLGA/鱼皮胶原共轭静电纺丝膜可用作GBR的屏障膜。     

载积雪草总苷同轴和混纺纳米纤维膜的制备和缓释性能评价

目的 采用静电纺丝法制备同轴及混纺载积雪草总苷的纳米纤维膜,研究其形貌及体外释药行为,为深Ⅱ度烧伤提供新型功能性敷料.方法 采用静电纺丝工艺,制备芯层载积雪草总苷的同轴海藻酸钠／聚乙烯醇-壳聚糖纳米纤维膜.进行处方工艺优化,包括壳层、芯层中聚合物浓度、制备工艺等,同时制备海藻酸钠／聚乙烯醇混纺载积雪草总苷纳米纤维膜.用扫描电子显微镜、透射电子显微镜和X射线衍射表征纳米纤维.考察载积雪草总苷混纺和同轴纳米纤维膜的体外释药行为.结果 载积雪草总苷同轴纳米纤维膜的最优处方是壳层由0.8％海藻酸钠、7％聚乙烯醇混合溶液制备,芯层由含2.5％积雪草苷的3％壳聚糖溶液制备.最优工艺是静电电压23 kV,针头和接收屏距离15 cm,壳层溶液推注速度0.8 ml/h,芯层溶液推注速度0.2 ml/h.混纺纳米纤维膜与同轴纳米纤维膜的处方和工艺类似.同轴纳米纤维膜有明显壳核结构,其芯层直径为99.2nm,整个纤维直径为166.8 nm.与混纺纳米纤维比较,同轴纳米纤维膜有药物缓释效果,机制以Fick's扩散为主.结论 同轴纳米纤维膜有明显药物缓释效果,有利于积雪草苷发挥促愈合效果,可用于深Ⅱ度烧伤的治疗.     

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。     

聚乙烯醇/壳聚糖载药电纺纤维膜的制备及性能表征

目的：使用静电纺丝技术制备载不同含量丹参素钠的聚乙烯醇/壳聚糖载药电纺纤维膜。 方法：以聚乙烯醇(PVA)和壳聚糖(CS)为载体,丹参素钠(SAS)为模型药物,制备载药电纺纤维膜;借助扫描电镜观察纤维形貌,利用红外光谱分析其成分,用紫外可见吸收光谱仪检测纤维膜含药量,绘制药物释放曲线。 结果：不同含量丹参素钠的聚乙烯醇/壳聚糖载药电纺纤维膜纤维直径均在280~390 nm 之间,纤维膜含药量高,并能较好地实现丹参素钠的缓控释放。 结论：研究制备成功的聚乙烯醇/壳聚糖载药电纺纤维膜制备工艺简单,载药均匀,有明显的缓释性,可为皮肤局部给药系统的研究提供新策略。     

三种羧甲基壳聚糖的初级生物学评价

本文旨在对三种不同取代位置的羧甲基壳聚糖（N-羧甲基壳聚糖（CMC）,N,O-羧甲基壳聚糖,O-羧甲基壳聚糖）进行初级生物学评价。本文利用化学改性的方法制备的三种羧甲基壳聚糖,其生物学评价试验包括细胞毒性试验和溶血试验。细胞毒性试验：采用浸提液试验中的细胞增殖度法提取材料浸提液培养L929细胞,观察其对细胞生长、增殖的影响,测定加样后2、4、7d的OD值,计算RGR,评定细胞毒性等级;溶血试验：采用溶血率法评价材料的溶血性能,取新鲜兔血与材料直接接触,测定溶血率。毒性试验结果发现：经纯化的壳聚糖和三种CMC的细胞毒性均为0级或1级,符合医疗用途的技术要求。材料浸提液浓度越大,细胞毒性越大,壳聚糖的三种羧甲基化产物中,N-CMC和N,O-CMC的细胞毒性比壳聚糖小,O-CMC的细胞毒性最大。溶血试验结果发现：壳聚糖和三种CMC的溶血率均小于5%,符合医疗用途的技术要求。N-CMC的溶血率最小,O-CMC的最大,这结果与细胞毒性试验一致。经体外细胞毒性试验和溶血试验可知：经初步检验,可认为壳聚糖的羧甲基化衍生物具有良好的生物相容性,为以后更深一步的研究提供了科研基础和实验依据。     

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.     

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.     

Electro-Acupuncture Combined with the Trigger Point Needle-Embedding for Treatment of Primary Trigeminal Neuralgia in 31 Cases

In recent years, the author of this essay has applied electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases, yielding satis- factory results as reported in the following.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Persist in and Carry Forward the Integrative Medical Research——Speech at the 6th National General Congress of Chinese Association of Integrative Medicine by Academician CHEN Zhu,Minister of Ministry of Health

The 6th National General Congress of Chinese Association of Integrative Medicine （CALM） was convened at 19-20, April 2008 in Beijing. Academician CHEN Zhu, the minister of Ministry of Health indicated at the congress that the integration of Chinese and Western medicine is very well in keeping with the situation of our country and the general rule of development in medical science; and as a good integration of Chinese medicine and Western medicine, it is mutually beneficial and advantageous to both of them. Seeing the creativity shown in integrative medical investigation in theoretic and methodological sides, we should and must persist in and develop it.