SD大鼠骨髓基质细胞与多孔羟基磷灰石陶瓷支架体外粘附的实验研究

目的研究诱导培养的SD大鼠骨髓基质细胞(bone marrow stromal cells,BMSCs)在多孔羟基磷灰石陶瓷支架材料上的粘附和生长。方法SD大鼠骨髓基质细胞(bone marrow stromal cells,BMSCs)经矿化诱导培养、扩增后与多孔羟基磷灰石陶瓷支架体外复合培养,扫描电镜观察BMSCs在支架表面的贴附形态;同时以不同浓度细胞悬液接种多孔羟基磷灰石陶瓷支架材料,检测适宜的接种浓度及单位体积支架材料最多可粘附BMSCs数量。结果BMSCs在支架表面贴附良好,当接种浓度为2×106/ml时,单位体积的多孔羟基磷灰石陶瓷支架材料最多可粘附BMSCs数量为3.1×107cell/cm3。结论SD大鼠BMSCs在体外经矿化诱导培养可表达成骨细胞表型,诱导培养后的细胞与新型多孔羟基磷灰石陶瓷支架材料上具有良好的亲和能力。     

BG/PHBV复合多孔支架材料与成骨细胞体外复合培养

目的用生物活性玻璃BG/聚羟基烷酸酯PHBV复合多孔支架材料与骨髓基质细胞来源的成骨细胞体外复合培养了解其生物相容性,为骨组织工程支架材料选择提供依据。方法:将体外培养的兔骨髓基质细胞诱导分化的成骨细胞,与BG/PHBV复合多孔支架材料体外复合培养,对复合体进行形态学、扫描电镜、细胞增长能力的测定,评价细胞与材料的相容性。结果:骨髓基质细胞有较强的向成骨细胞分化和繁殖能力成骨细胞与BG/PHBV复合多孔支架材料体外复合培养8d,分布于支架材料的成骨细胞迅速分化增殖,分泌细胞外基质并形成钙结节。结论:骨髓基质细胞是骨组织种子细胞的良好来源BG/PHBV复合多孔支架材料与成骨细胞具有良好的生物相容性,是构建组织工程骨的一种理想支架材料。     

老年人牙周膜细胞附着于纳米-羟基磷灰石上的形态学观察

目的:老年人牙周膜细胞的再生能力相对较低,选择合适的支架材料很重要.文中观察接种在三维多孔纳米-羟基磷灰石支架材料上的老年人牙周膜细胞组织形态学表现. 方法:将原代培养的老年人牙周膜细胞接种于三维多孔纳米-羟基磷灰石上,在培养15d后,采用扫描电子显微镜观察细胞的生长情况,并与青年人牙周膜细胞进行比较. 结果:老年人牙周膜细胞接种到支架材料上15d后表现为星形或多角形,有良好伸展性和对支架材料的黏附性,一定量的钙结节生成,但产生的钙结节量较青年人牙周膜细胞少. 结论:老年人牙周膜细胞可以在三维多孔纳米-羟基磷灰石支架材料上良好生长.三维多孔纳米-羟基磷灰石支架材料可以作为老年人牙周组织工程治疗中的支架材料.     

大鼠骨髓间质干细胞体外分化为成骨细胞的实验研究

目的：建立大鼠骨髓间质干细胞(mesenchymal stem cells，MSCs)体外诱导分化为成骨细胞的模型。方法：分离大鼠骨髓间质干细胞进行体外培养，观察其生物学特性。并选用一定的诱导剂诱导成骨细胞，通过形态学变化、碱性磷酸酶染色及钙沉积对成骨细胞进行鉴定。结果：大鼠MSCs细胞形态呈长梭形，成骨细胞诱导后MSC细胞形态由长梭形向多边形转变，ALP染色阳性，Von Kossa染色阳性，呈现典型的成骨细胞形态和生物学特征。结论：大鼠骨髓MSCs体外能被诱导分化为成骨细胞，为骨组织工程研究的优良种子细胞。     

人骨髓间充质千细胞与骨转化羟基磷灰石复合培养研究

目的 观察人骨髓间充质干细胞(Mesen chymal stem cell，MSC)与骨转化羟基磷灰石的复合培养，证实骨转化羟基磷灰石是组织工程良好的载体材料。方法制备骨转化羟基磷灰石，体外培养MSC并材料复合，通过共聚焦显微镜和环境扫描电镜观察MSC与材料的复合情况。结果 MSC可以在材料上贴附、生长、繁殖、分化。结论 骨转化羟基磷灰石是良好的支架材料，易与MSC复合，适合种子细胞的贴附、生长、增殖和分化。     

兔骨髓源成骨细胞与多孔支架复合的实验研究

目的通过对兔骨髓基质干细胞的培养及诱导分化,并将细胞与新型复合型生物多孔支架联合培养,观察兔骨髓源成骨细胞在多孔支架表面的黏附及生长情况。方法兔骨髓基质干细胞经诱导分化为成骨细胞。将成骨细胞与处理过的多孔支架复合,在扫描电镜下观察细胞在支架表面的贴附情况。结果细胞接种后,在材料表面生长形态正常,属性与特征明显。结论新型复合型生物多孔支架与兔骨髓源成骨细胞具有良好的结合能力,可以进一步进行体内试验。     

鸵鸟羟基磷灰石陶瓷支架负载骨髓基质细胞异位成骨性能

目的:观察鸵鸟羟基磷灰石陶瓷支架负载骨髓基质细胞后植入裸鼠皮下的成骨性能. 方法:获取兔髂骨松质骨骨髓基质细胞,体外分离、扩增、诱导后接种于鸵鸟羟基磷灰石支架. 支架/细胞复合物植入裸鼠背部皮下,支架单纯植入作为对照. 植入后3, 6 wk取材,通过大体、组织学观察评价成骨活性. 结果:支架/细胞复合物植入后3 wk,以软骨为主的大量未成熟骨在材料表面及孔隙内形成;植入后6 wk,更多的成熟骨形成. 在支架材料和骨组织的邻接区域见成骨细胞及破骨细胞,部分区域有血管和多核巨细胞分布,可见软骨内成骨方式. 结论:支架/细胞复合物显示良好的成骨活性,鸵鸟羟基磷灰石陶瓷可以用于骨组织工程支架材料.     

骨髓来源成骨细胞复合纳米晶羟基磷灰石胶原构建组织工程骨的实验研究

目的 :探讨纳米材料作为组织工程骨基质材料的可行性。方法 :分离培养兔骨髓间充质干细胞 ,诱导为成骨细胞后作为种子细胞 ,与纳米晶羟基磷灰石胶原材料于体外联合培养 ,复合物植入兔桡骨节段性缺损处 ,通过大体观察、HE染色及X线摄片了解成骨情况。结果 :兔骨髓间充质干细胞在体外可以大量扩增 ,能定向诱导为成骨细胞 ;复合物植入 16周后 ,X线摄片中可见桡骨缺损处连接良好。结论 :纳米晶羟基磷灰石胶原材料是组织工程骨的较好的支架材料。     

胶原-羟基磷灰石-硫酸软骨素-骨形态发生蛋白骨修复材料的性质评估

目的：评估以胶原、羟基磷灰石、硫酸软骨素等3种天然骨骼基本成分构建成的三维多孔骨修复材料的理化性能和体内生物学性能,观察其作为骨形态发生蛋白（bone morphogenetic protein 2,BMP-2)载体的效果。方法：以胶原、羟基磷灰石、硫酸软骨素及BMP-2为原料,通过化学交联和冷冻干燥的方法构建具有一定三维结构的骨修复材料。通过HE染色、扫描电镜观察材料的结构性能;通过表面能谱、X线衍射观察材料的理化性能;将骨髓基质干细胞（marrow stromal cells,MSCs）种植在材料表面,观察MSCs在材料表面的粘附、增生和分化;将该复合材料种植在大鼠体内,观察材料在体内的降解和异位成骨情况。结果：骨修复材料在植入局部保持完整的支架结构,具有利于细胞粘附和增殖的多孔结构。通过肌肉埋植,在异位诱导形成了骨组织,并且随着骨组织的形成,支架逐渐降解吸收。结论：胶原 羟基磷灰石 硫酸软骨素 骨形态发生蛋白是具有良好的生物相容性和骨诱导特性的骨修复材料。     

MMT法评价多孔纳米羟基磷灰石/壳聚糖支架材料的细胞毒性

目的制备多孔纳米羟基磷灰石/壳聚糖（nHA/CS）,对其进行细胞毒性检测。方法采用共沉淀法和粒子沥滤法制备多孔nHA/CS。分离培养大鼠成骨细胞,碱性磷酸酶（ALP）染色进行成骨细胞进行鉴定,倒置显微镜下观察培养过程中细胞的形态和增殖情况。MTT检测多孔nHA/CS支架材料对成骨细胞增殖的影响,评价细胞毒性。结果细胞可在多孔nHA/CS周围贴壁生长,并且附着、增殖,其生长及功能不受抑制。结论多孔nHA/CS无细胞毒性,有利于细胞的黏附和增殖,可以用于骨组织工程支架材料。     

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.     

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.