骨髓问充质干细胞对大鼠损伤汗腺的修复性研究

目的探讨皮肤汗腺受损后骨髓间充质干细胞对其的修复效果。方法大鼠骨髓间充质干细胞体外分离、培养、标记。机械损伤大鼠脚掌汗腺,干细胞治疗组于创面周围皮下注入MSCs,对照组注射相同剂量PBS,分别比较两组的创面愈合率及再生上皮的厚度,观察MSCs参与受损汗腺修复的情况。结果7d时干细胞治疗组的创面愈合率及14d时的表皮再生厚度均优于对照组（P〈0．05）;BrdU标记的MSCs参与了损伤汗腺导管部的修复。结论骨髓间充质干细胞有利于创伤皮肤的修复并可能参与损伤汗腺的修复。     

骨髓间充质干细胞修复兔皮肤缺损创面的实验研究

目的研究骨髓间充质干细胞(MSCs)参与皮肤缺损创面修复重建表皮的可能性,为组织工程化皮肤提供新的种子细胞来源。方法自体来源的骨髓MSCs应用5-溴脱氧尿嘧啶(BrdU)标记后,以Ⅰ型胶原为载体植入兔背部左侧全层皮肤缺损创面(治疗组),右侧全层皮肤缺损创面仅植以Ⅰ型胶原作为对照组。于术后6、12天观察创面收缩率,记录愈合时间;术后4、5周切取创面中央再生皮肤组织,行常规病理和BrdU免疫组织化学染色检测,进行对比研究。结果创面收缩率,治疗组>对照组(P<0.05);愈合时间,治疗组<对照组(P<0.05)。常规病理检测治疗组再生皮肤表皮层明显增厚,细胞数目显著增多;免疫组化检测,在治疗组再生皮肤附件毛囊内层以及表皮基底层、棘层可见BrdU阳性标记细胞。结论骨髓间充质干细胞在全层皮肤缺损创面可能分化为表皮细胞参与表皮重建并具有促进创面愈合的作用。     

MSCs移植并GS对大鼠脊髓损伤后期修复的影响

目的观察骨髓间充质干细胞(MSCs)移植联合应用促细胞生长物质(GS)对脊髓损伤后期大鼠运动功能修复的影响。方法应用全骨髓法分离培养MSCs,10 mg/L的BrdU标记细胞核。将成年雄性Wistar大鼠36只,以改良Allen打击法制备T10脊髓损伤模型,制模后2周随机分为MSCs+GS组、MSCs组与对照组,每组12只,伤后2周各组损伤处分别注入MSCs+GS、单纯MSCs、培养液。分别于伤后1、2、3、4、5、6周进行BBB评分;损伤后6周处死大鼠,取损伤段脊髓及其上下各1 cm组织,行苏木精-伊红染色及SABC免疫组织化学方法染色,观察损伤脊髓组织病理变化和BrdU阳性细胞及GAP-43的相对表达。结果制模后1~2周3组大鼠后肢运动功能BBB评分未见明显差异(F=0.322、0.044,P>0.05),3~6周MSCs+GS组大鼠后肢运动功能BBB评分较MSCs组和对照组高(F=13.729~97.187,P<0.05);MSCs+GS组大鼠脊髓损伤中心及头、尾端均可见BrdU染色阳性细胞。同时,MSCs+GS组及MSCs组损伤节段脊髓内GAP-43的表达面积明显多于对照组,MSCs+GS组多于MSCs组。结论 MSCs移植联合GS促进大鼠损伤脊髓结构和功能恢复的效果明显优于单纯细胞移植,两者联用具有协同效应。     

羊膜载体复合膜负载骨髓间充质干细胞促愈创面的研究

目的 探讨骨髓间充质干细胞(MSCs)促进创面愈合的作用和用于创面修复的可行性,为组织工程化皮肤提供新的种子细胞来源.方法 自体来源的骨髓,经体外培养并纯化MSCs,应用5-溴脱氧尿嘧啶(BrdU)标记技术进行标记,以羊膜载体复合膜为载体植入兔背部左侧全层皮肤缺损创面(治疗组),右侧全层皮肤缺损创面仅植以羊膜载体复合膜(对照组).于术后1,2周观察创面收缩率,记录愈合时间;术后4,5周切取创面中央再生皮肤组织,行病理和BrdU免疫组织化学染色检测,进行比较.结果 创面收缩率,治疗组>对照组(P<0.05),愈合时间,治疗组<对照组(P<0.05).常规病理检测治疗组再生皮肤表皮层明显增厚,细胞数目显著增多.免疫组化检测,在治疗组再生皮肤附件毛囊内层以及表皮基底层可见BrdU阳性标记细胞.结论 MSCs自体移植在一定程度上可以显著提高创面愈合的质量,为皮肤损伤由解剖重建到功能修复的完美愈合提供了新的治疗思路.     

间充质干细胞经鼻内途径移植对创伤性脑损伤的精准修复作用

目的：探讨间充质干细胞（MSCs）经鼻内途径移植对创伤性脑损伤（TBI）的定向精准修复作用。方法：75只雄性SD大鼠按随机数字表法分为假手术对照组、模型对照组、患处MSCs移植组、鼻内MSCs移植组、尾静脉MSCs移植组，每组15只。采用改良式自由落体法制备中度TBI模型，分别以患处、鼻内、尾静脉途径移植MSCs(5×105个/只)。移植后第1、第7天通过改良神经功能缺损评分（m NSS）了解大鼠神经功能恢复情况；解剖观察脑组织恢复情况；比较各组脑脏器系数；普鲁士蓝染色观察超顺磁性纳米铁颗粒（SPIO）标记的MSCs经鼻内途径入颅情况；HE染色观察各组大鼠脑组织病理学改变情况。结果：m NSS结果显示，模型对照组术后第1、第7天mNSS评分明显高于假手术对照组（P<0.01）。与模型对照组比较，MSCs移植各组术后第1、第7天m NSS均显著降低（P<0.01）。解剖观察显示，与模型对照组比较，移植后第7天MSCs移植各组大鼠脑创面均有明显改善。与假手术对照组比较，移植后第1天模型对照组脑脏器系数显著升高（P<0.05）；与模型对照组比较，患处及鼻内MSCs移植组脑脏...     

骨髓间充质干细胞促进大鼠皮肤创面愈合的作用及机制研究

目的 探讨骨髓间充质干细胞(mesenchymal stem cells,MSCs)对大鼠皮肤创面的促愈作用及机制,为临床皮肤损伤修复提供新的治疗方法.方法 建立大鼠背部皮肤全层切割伤模型,将大鼠分为2组:空白对照组,MSCs处理组,分别于伤后3、6、9、12 d采用大体观察,常规组织学检查观察2组大鼠背部皮肤伤口愈合状况及RT-PCR观察转化生长因子(TGF-β1)、表皮生长因子(EGF)mRNA在愈合中的变化特点.结果 MSCs处理组大鼠皮肤愈合快,愈合指数高,创伤后第3天MSCs处理组大鼠皮肤TGF-β1上调明显,第6天EGF上调明显.结论 MSCs能促进大鼠皮肤创面愈合,其机制可能与其引起各种生长因子的合成和分泌有关.     

脑梗死后自体神经干细胞原位增殖与分化的实验研究

目的研究成年大鼠脑梗死后自体神经干细胞的原位增殖和分化。方法雄性Wistar大鼠共112只,分对照组(12只)、脑梗死后1d组(20只)、脑梗死后3d组(20只)、脑梗死后7d组(20只)、脑梗死后14d组(20只)、脑梗死后28d组(20只)。大鼠于不同的时间处死,并取出大脑,处死前均注射5-溴胱氧腺苷嘧啶(BrdU)。用免疫组织化学方法动态检测BrdU、Musashil、胶质纤维酸性蛋白(GFAP)、神经元核性蛋白(NeuN)的表达。BrdU、Musashil确定神经干细胞的增殖,GFAP、NeuN确定神经干细胞的分化。结果与对照组相比,大鼠海马BrdU^ 细胞和BrdU^ ／Musashil^ 细胞数在脑梗死后1d组开始增加,7d组达到高峰,28d组接近正常水平;BrdU^ ／GFAP^ 细胞数无明显变化;BrdU^ ／NeuN^ 细胞数在脑梗死后14d组开始增加,28d组最多。结论大鼠脑梗死激活自体神经干细胞原位增殖,并且大多数增殖的神经干细胞分化成神经元。     

表皮干细胞复合羊膜构建组织工程皮肤修复糖尿病难愈创面的可行性分析

目的对表皮干细胞复合羊膜构建组织工程皮肤修复糖尿病难愈创面的可行性进行分析和探讨。方法选取60只体质量250~300 g的雄性SD大鼠,建立糖尿病SD大鼠创面模型,分离培养大鼠皮肤表皮干细胞(ESCs),使用5-溴脱氧尿嘧啶(5-Brd U)对其进行标记。使用随机数表法将大鼠分为A、B、C组,A组大鼠皮肤创面接受羊膜负载Brd U标记的ESCs组织工程皮肤移植;B组大鼠皮肤创面接受羊膜移植;C组大鼠皮肤创面不接受处理。观察3组大鼠皮肤创面的愈合情况。结果 A组大鼠治疗7、14 d后的PCNA阳性细胞积分光密度平均值以及创面愈合率均高于B、C组,差异有统计学意义(P<0.05)。结论表皮干细胞复合羊膜构建组织工程皮肤修复糖尿病难愈创面安全可行。     

永生化神经前体细胞治疗大鼠局灶性脑缺血的实验研究

目的 探讨永生化神经前体细胞(INPC)移植对局灶性脑缺血大鼠神经功能的影响及移植细胞在模型大鼠脑内的存活以及分化情况.方法 雄性SD大鼠24只,均采用线栓法建立大脑中动脉缺血(MCAO)模型,随机分为2组(每组12只):脑缺血对照组、INPC移植组.缺血后3 d通过立体定位注射方法分别将等体积的磷酸盐缓冲液、5-溴脱氧尿嘧啶核苷(BrdU)标记的INPC悬液注射到2组大鼠纹状体缺血半暗带区.缺血再灌注后对大鼠进行神经损害严重程度评分(NSS).细胞移植后1和4周分别随机处死2组大鼠各6只,取脑组织制作冰冻切片,通过免疫荧光双标技术检测BrdU、胶质纤维酸性蛋白(GFAP)双阳性细胞和BrdU、神经元特异性烯醇化酶(NSE)双阳性细胞,观察INPC在移植区域的存活及分化情况.结果 脑缺血对照组与INPC移植组比较,细胞移植前后各时点的NSS评分差异均无统计学意义(均P＞0.05).INPC移植组细胞移植后1及4周,均可在移植针道附近及缺血灶周围检测到聚集较明显的BrdU免疫荧光染色阳性的植入细胞,并观察到BrdU染色阳性细胞扩散至全脑,免疫荧光双标技术检测见部分细胞为BrdU、GFAP双阳性的星形胶质细胞或BrdU、NSE双阳性神经元.结论 INPC可在局灶性脑缺血大鼠脑内存活并分化为神经元和星形胶质细胞.     

骨髓间充质干细胞自体移植提高猪皮肤创面修复质量的初步研究

目的　探讨骨髓间充质干细胞 (MSCs)局部移植对提高猪皮肤烫伤创面组织修复质量的作用 ,为临床皮肤损伤后功能性修复提供新的治疗方法。方法　抽取 6只小型香猪的骨髓 ,经体外培养并纯化MSCs,应用 5 溴脱氧尿嘧啶 (BrdU)标记技术进行标记。猪背部皮肤中线两侧各制备 6个面积为 2 5 4cm2 的深Ⅱ0 烫伤创面 ,将已标记的MSCs( 2× 10 6)以注射方式回植到提供骨髓猪的创面下 ,将创面随机分为 6组 ,即空白对照组、MSCs治疗组、MSCs 碱性成纤维细胞生长因子 (bFGF)治疗组、bFGF治疗组、MSCs 表皮细胞生长因子 (EGF)治疗组以及EGF治疗组。分别于伤后 7、14、2 1、4 2d采用大体观察、常规组织学检查、免疫组织化学及免疫荧光化学染色动态观察创面愈合情况。结果　实验猪皮肤烫伤后 7d开始 ,各组创面逐渐缩小 ,伤后 2 1d ,大部分创面愈合。虽然不同时间点各治疗组创面面积缩小率差异无显著意义 ,但以MSCs bFGF治疗组最为明显 ,伤后第 14天和 2 1天创面缩小的面积比其他 5组大 15 %～ 2 0 %。半定量评价结果显示MSCs bFGF治疗组肉芽组织中血管密度较大 ,为 ,而其他组别仅为 ～ 。再上皮化的新生表皮在MSCs bFGF治疗组较其余 5组厚 ,并有上皮角形成。半定量分析可见神经纤维在MSCs bFGF治疗组较其他 5     

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.