两种骨髓间充质干细胞移植途径治疗急性心肌梗死的比较研究

目的:探讨两种干细胞移植方式对心肌梗死后心功能的影响,为临床治疗急性心肌梗死探求更为理想的干细胞移植途径.方法:结扎法建立小鼠心肌梗死模型,然后将造模成功的小鼠分为心肌移植组、静脉移植组和对照组.体外培养扩增、纯化骨髓间充质干细胞,移植组干细胞分别经尾静脉和心肌内注射的方式移植入模型小鼠体内,对照组经尾静脉注射等量PBS溶液.2周后超声心动图评价心功能及病理学检测梗死面积.结果:2周后超声心动图心功能检测3组左室射血分数分别是(26.58±5.1)%、(16.90±3.4)%、(17.02±3.34)%;梗死面积分别为(52.15±4.22)%、(56.95±4.10)%、(57.12±3.89)%.结果显示,直接心肌内注射移植干细胞能够减少梗死面积,显著改善心脏功能;静脉移植方式未能改善心脏功能.结论:细胞移植方式是影响心脏干细胞治疗效果的重要因素,直接心肌内骨髓间充质干细胞移植途径在保护心功能方面优于静脉移植.     

Y染色体追踪大鼠骨髓间充质干细胞心肌移植的实验研究

目的 利用Y染色体检测雄性大鼠的骨髓间充质干细胞(MSCs)移植到雌性大鼠心肌梗死模型中的存活情况。方法 采用全骨髓贴壁筛选法体外培养雄性大鼠的骨髓间充质干细胞作为移植细胞,选取80只雌性大鼠通过结扎左冠状动脉前降支制备心肌梗死模型,其中45只成功建立模型。成模后2周按以下分组进行细胞移植：将模型动物随机分为心外膜直接注射MSCs组(A组,n=15)、经静脉移植MSCs组(B组,n=15)和经静脉注射等量DMEM培养液组(C组,n=15),通过实时定量PCR检测雄性大鼠Y染色体特有的SRY基因,观察移植后1d、1周、2周及3周时MSCs在梗死心肌的存活情况。结果 经心外膜直接注射组和经静脉移植组通过PCR均可检测出SRY基因的存在,移植后两组的心功能得到明显的改善(P<0.05)。结论 MSCs经心外膜和静脉移植均可分布于心肌梗死周围区域,对梗死后大鼠的心脏具有保护作用。     

液态心肌细胞外基质心肌内注射改善大鼠急性心肌梗死后心功能的实验研究

目的 探讨对心肌梗死区注射液态心肌细胞外基质能否改善大鼠急性心肌梗死后的心功能.方法 制备液态心肌细胞外基质,采用开胸结扎的方法制备大鼠急性心肌梗死模型,将制备成功的模型随机分为3组:Ⅰ组为液态心肌细胞外基质组(n=25),Ⅱ组为骨髓间充质干细胞组(n=25),Ⅲ组为生理盐水对照组(n=25),并对各组予以相应处理.在...     

骨髓间充质干细胞联合促红素治疗大鼠急性心肌梗死实验研究

目的:观察静脉移植骨髓间充质干细胞(MSC)联合促红素(EPO)治疗心肌梗死的可能性。方法:大鼠MSC培养后,制备成悬液备用。40只Lewis大鼠用异丙肾上腺素(ISO)制作急性心肌梗死模型,随机分为4组:A.MSC加EPO治疗组(n=10);B.MSC治疗组(n=10);C.EPO治疗组(n=10);D.心肌梗死对照组(n=10)。4周后,观察大鼠的心功能、血流动力学变化。结果:与对照组相比,MSC加EPO组、MSC组和EPO组的心功能明显改善,左室腔明显缩小(P<0.05),梗死面积减少;其中MSC加EPO组较MSC组和EPO组的心功能好(P<0.05)。结论:骨髓MSC联合EPO治疗大鼠急性心肌梗死取得了良好的效果,比单用效果好。     

神经干细胞与骨髓间充质干细胞移植对脊髓损伤修复的比较

目的:比较神经干细胞和骨髓间充质干细胞移植治疗脊髓损伤效果与机制的差异。方法:用成年Wist-ar大鼠建立脊髓半横切模型,随机分为神经干细胞(NSCs)组(n=10)、骨髓间充质干细胞(BMSC)注射组(n=10)、PBS注射组(n=10)及只打开椎板的假手术组(n=10)。移植后行BBB运动功能学评分,半切位置的免疫组化及核磁成像比较。结果:BBB评分NSCs注射组明显高于BMSCs注射组,NSCs注射组和BMSCs注射组均明显高于PBS注射组,脊髓切片中可观察到被标记的NSCs及BMSCs,核磁成像显示移植后半切形成的脊髓空洞有所减小。结论:静脉注射NSCs、BMSCs均能改善脊髓损伤大鼠的运动功能,但注射NSCs效果更明显。静脉注射干细胞后,细胞可迁移至脊髓损伤的位置,核磁成像显示脊髓空洞有所减小,提示干细胞可能通过补充或替代损失的神经细胞,修复已缺失的神经组织和功能性神经单位,重建神经环路。     

芪参益气滴丸联合骨髓间充质干细胞移植对心肌梗死小鼠血管新生及心功能的影响研究

目的:利用体外细胞分子生物学实验探讨芪参益气滴丸联合骨髓间充质干细胞移植对心肌梗死后小鼠血管生成及心功能的影响.方法:采用C57BL/6J的雄性小鼠,将60只小鼠采用数字随机法分为3组(n=20):心肌梗死组(MI+PBS组)、心肌梗死+间充质干细胞移植组(MI+MSCs组)和心肌梗死+间充质干细胞移植+芪参益气滴丸组...     

骨髓间充质干细胞移植对心肌梗死大鼠损伤心肌的修复效果

目的 探讨骨髓间充质干细胞(BMSCs)移植对心肌梗死大鼠损伤心肌的修复效果,为进一步提高BMSCs移植治疗心肌梗死的疗效提供实验依据.方法 将22只大鼠参照Olivette方法制作心肌梗死模型,共16只进入实验终点,随机分为2组:对照组(C组,n=8)梗死区注射P BS;骨髓间充质干细胞移植组(M组,n=8)梗死区注射BMSCs.术后喂养4周,行心脏彩超,取心脏组织,荧光显微镜观察BMSCs的分布、分化及梗死区毛细血管密度.结果 标记CM-Dil和cTnT免疫组织化学染色双重阳性的心肌样细胞在梗死区分布.与C组相比,M组大鼠左室收缩末期内径(LVEDd)、左室舒张末期内径(LVEDs)、射血分数(EF)、左室短轴缩短率(FS)均有明显改善(P＜0.05),毛细血管密度明显增加(P＜0.05).结论 BMSCs可在心肌梗死的大鼠心脏分化为心肌样细胞,提高梗死区毛细血管密度,改善心功能.     

同种异体骨髓间充质干细胞在大鼠心脏的迁移及分化

目的探讨同种异体骨髓间充质干细胞(MSC)在大鼠心脏定居存活情况、同种异体移植是否可行。正常心脏微环境与梗死心脏微环境对骨髓间充质干细胞(MSC)迁移、分化的影响。方法雌性Wistar大鼠35只,①正常心脏MSC移植组(10只);②急性心肌梗死对照组(AMI,10只);③心肌梗死MSC移植组(10只);④单个核细胞治疗组(5只)。结扎左冠状动脉前降支造成心肌梗死模型。分离纯化来自雄性Wistar大鼠的骨髓MSC,于冠脉结扎后1～3h植入到雌性正常大鼠和心肌梗死大鼠心脏组织,模拟同种异体细胞移植治疗。于细胞移植后10周取心脏检测各种指标。结果(1)经纯化的大鼠MSC在同种异体大鼠心脏可定居、生存,而未经纯化的单个核细胞移植在同种异体大鼠心脏未见存活;(2)在正常心脏微环境中,带蓝色荧光的供体MSC细胞核呈小岛屿状分布,与宿主心肌纤维排列无关;而在心肌梗死模型大鼠心脏微环境中,蓝色细胞核分布广泛,呈椭圆形,似心肌细胞核,其排列方向与宿主心肌纤维排列方向一致,免疫组化检测胞浆心肌特异蛋白染色阳性。结论纯化的MSC具有免疫耐受特性,无需加用免疫抑制剂,可进行同种异体移植;同种异体MSC在正常心脏微环境中不发生迁移、分化;而在急性梗死大鼠心脏中具有向缺血梗死区迁移并分化为心肌样细胞的特点。     

二维应变超声心动图评价干细胞移植对心肌梗死大鼠左心室局部收缩功能的影响

目的应用二维应变超声心动图(2-dimensional strain echocardiography,2DSE)观察心肌梗死大鼠在骨髓间充质干细胞(mesenchymal stem cells,MSCs)移植后局部心肌功能改善情况,探讨2DSE在心肌梗死大鼠干细胞移植研究中心功能评价的价值。方法 40只SD大鼠随机分为心肌梗死组(n=14)、MSCs治疗组(n=16)和对照组(n=10),心肌梗死组与MSCs组建立心肌梗死模型,对照组只进行假手术。MSCs组在梗死区心肌局部注射DAPI标记的MSCs,心肌梗死组和对照组只注射DMEM培养基。分别于术前1天和术后1周、术后4周用2DSE观察左心室乳头肌水平短轴局部室壁运动情况。术后4周用荧光显微镜观察心肌梗死区移植细胞分布,免疫组化检测移植细胞的肌钙蛋白表达情况。结果术后4周,MSCs治疗组局部室壁的径向应变和应变率高于心肌梗死组(P<0.01);MSCs组心肌梗死区可见DAPI标记阳性的细胞,该区肌钙蛋白表达阳性。结论 2DSE可以检测出心肌梗死大鼠在MSCs移植后局部心肌收缩功能的改善,可成为干细胞移植研究中可选择的敏感的无创性指标。     

移植时机对骨髓间充质干细胞修复梗死心肌的影响

目的:观察心肌梗死后不同炎症微环境对骨髓间充质干细胞(MSCs)移植后在大鼠梗死心肌中的定位、存活及分化情况,探讨细胞治疗的最佳时间窗. 方法:建立大鼠左冠状动脉结扎的心肌梗死模型,心肌梗死后1 h及1,2,4,8 wk分别取心脏进行组织学观察;将经过4',6-二脒-2-苯基吲哚(4',6-diamidino-2-phenylindole, DAPI)标记的同种骨髓间充质干细胞在心肌梗死后1 h及1,2,4,8 wk移植到心肌梗死边缘区(每组n=10),对照组心肌梗死后1 h注射等量培养基(n=10). 心肌梗死后不同时间点采集心脏标本进行HE染色观察炎症反应的变化,细胞移植治疗各组在心肌梗死后8 wk测定心脏功能、心脏瘢痕面积以及免疫组化染色鉴定细胞分化情况. 结果:分离纯化的大鼠MSCs经流式细胞术鉴定为CD44 CD90 /CD34-CD45-CD31-,心肌梗死后1 wk梗死相关区的炎症反应较强,2 wk时减弱, 4～8 wk时瘢痕完全形成并出现左室重塑. 1 wk移植组未发现有移植细胞存活, 左室心功能及瘢痕面积百分比与对照组相似,1 h及2,4 wk细胞移植组心肌组织冰冻切片中可观察到大量标记细胞存在,并向周围迁移分布,免疫组化显示阳性标记细胞desmin, ctnI 阳性,左室血流动力学指标及瘢痕面积百分比与对照组比较均有改善(P<0.05), 2 wk的左室收缩末压力、左室最大上升速率及瘢痕面积百分比的改善优于1 h组和4 wk移植组(P<0.05). 结论:心肌梗死后炎症反应强烈, 不利于移植细胞的存活,而在炎症减弱的瘢痕修复早期尽早进行MSCs移植可能对于细胞的存活及心功能的恢复较为有利.     

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.     

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.     

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.