人外周血树突状细胞的诱导及其生物学特性的研究

目的：建立体外诱导和扩增人外周血树突状细胞（ＤＣ）的方法,并分析其生物学特性。方法：造血动员后外周血单个核细胞经贴壁去除悬浮细胞,加入细胞因子（ＩＬ－４,ＧＭ－ＣＳＦ和ＴＮＦ＿α）培养８８天。用流式细胞仪分析细胞的表型,经ＥＬＩＳＡ法测定其培养上清中的ＩＬ－１２,并将诱导的树突状细胞与脐带血原始Ｔ细胞混合培养,用＾３Ｈ－ＴｄＲ掺入法测定细胞的增殖指数。结果：贴壁的造血动员后外周血单个核细胞在体外经     

细胞因子诱导不同来源的贴壁细胞分化为树突状细胞的研究

目的：探讨寻树突状态细胞（DC）体外培养的理想来源。方法：通过贴壁的方法分离脐带血、造血动员后的外周血及正常人外周血中DC的前体细胞，体外采用细胞因子诱导，并对其进行形态学的观察和细胞表型检测；同时比较了不同来源的DC的增殖、分泌IL－12的能力及激发同种异体T细胞和脐带血幼雅T细胞（naive Tcells)的作用，结果：经过造血动员后的外周血产生的DC数量和纯度均较高，而且能经体外细胞因子诱导为功能性DC。结论：造血动员后的外周血是较理想的DC来源。     

动员人外周造血祖细胞定向诱导树突状细胞

目的 探讨由动员人外周造血祖细胞体外培养扩增获得的树突状细胞（ＤＣ）的生物学特性。为临床应用肿瘤树突状细胞疫苗建立制备方法。方法 用ＣＳ－３０００血细胞分离机分离预经干细胞动员的患者外周血单个核细胞（ＰＢＭＣ）,去除淋巴细胞和单核细胞,阴性选择得到外周造血祖细胞。加入ｒｈＧＭ－ＣＳＦ和ｒｈＩＬ－４培养１０￣１４ｄ成为树突状细胞,然后进行形态学检测、细胞表型和Ｔ细胞刺激能力分析。结果 由外周造血祖细     

不同细胞因子对培养人外周血树突状细胞的影响

目的建立从人外周血分离、纯化、培养、扩增树突状细胞（ＤＣ）前体的方法，研究细胞因子对ＤＣ体外增殖、分化成熟的影响。方法人外周血经血细胞分离仪及Ｆｉｃｏｌ、Ｐｅｒｃｏｌ等不连续密度梯度离心获得的含ＤＣ前体细胞组分用重组人粒细胞／巨噬细胞集落刺激因子（ｈｒＧＭ－ＣＳＦ）培养或用ＧＭ－ＣＳＦ及白细胞介素－４（ＩＬ－４）联合培养；光镜及电镜观察及ＡＢＣ法免疫细胞化学染色。结果分离的ＤＣ前体经１周左右时间培养，细胞数量可扩增２０～３０倍，纯度达９０％以上，ＧＭ－ＣＳＦ与ＩＬ－４联合培养所得到的ＤＣ数量约为用ＧＭ－ＣＳＦ培养的１．５倍。ＤＣ具有典型的树枝状或裙褶状突起，并表达高水平的ＨＬＡ－ＤＲ，其ＣＤ１４、ＣＤ１９、ＣＤ３的表达均为阴性。结论用ＧＭ－ＣＳＦ和ＩＬ－４联合作用更能促进ＤＣ的体外扩增及分化。     

树突状细胞刺激γδT细胞扩增诱导干细胞移植时GVL研究

目的：研究γδＴ细胞的体外生存期,抗白血病活性及可能引起的移植物抗宿主反应（ＧＶＨＤ）,为自体及异体移植后输注γδＴ细胞诱导移植物抗白血病反应作前期基础研究。方法：对移植患者采集物用ＧＭ－ＣＳＦ＋ＩＬ－４进行培养,使其分化和扩增为树突状细胞（ＤＣ）;然后用ＤＣ和细胞因子ＩＬ－２刺激其ＣＤ４＾＋Ｔ细胞扩增后,协同ＤＣ共同刺激γδＴ细胞扩增,进而用ＭＴＴ比色法和ＣＦＵ－ＧＭ集落培养研究其功能。结果：γ     

乳腺癌细胞抗原和自身淋巴结树突状细胞对细胞因子诱导的杀伤细胞作用研究

目的：从乳腺癌患者淋巴结　中分离和定向诱导培养扩增　树突状细胞（ＤＣ），观察其经自身肿瘤细胞抗原冲击后对自身细胞因子诱导的杀伤细胞（ＣＩＫ）　杀伤活性的影响。方法：取手术切除肿瘤周围转移的淋巴结，提取单个核　细　胞，将贴壁生长的细胞用细胞因子基因重组粒巨集落生长因子（ｒｈＧＭ－ＣＳＦ）、基因重组白介　素４（ｒｈＩＬ－４）、基因重组α－肿瘤坏死因子（ｒｈＴＮＦ－α）联合诱导培养ＤＣ，然后用　自身肿瘤细胞抗原冲击ＤＣ并作用ＣＩＫ，最后检测ＣＩＫ杀伤３种靶细胞（自身乳腺癌细胞、Ｋ５６２和　ＳＧＣ－７９０１细胞）的活性。结果：乳腺癌患者转移淋巴结中的贴壁细胞，经　细胞因子联合诱导培养，ＤＣ含量可达１５．３％～３７．０％。经自身肿瘤抗原冲击的淋巴结ＤＣ活化的Ｃ　ＩＫ，杀伤自身肿瘤细胞活性达７１．３％，单纯ＣＩＫ为３７．０％，两者比较差异有显著性（Ｐ＜０　．０１）；而对Ｋ５６２和ＳＧＣ－７９０１细胞杀伤活性无明显差异。结论：肿瘤周围转　移淋巴结贴壁细胞在体外能定向诱导扩增出大量ＤＣ；ＤＣ经自身肿瘤抗原冲击后能明显提高ＣＩ　Ｋ对自　身肿瘤细胞的杀伤活性。     

快速分离肝癌患者外周血树突状细胞

目的探讨从肝癌患者外周血中大量快速分离树突状细胞（ＤＣ）有效方法。方法自肝癌患者外周血中分离出单个核细胞（ＰＢＭＣ）;ＰＢＭＣ与Ｇｍ－ＣＳＦ及ＩＬ－４共培养;检测培养前后ＤＣ表面ＨＬＡ－ＤＲ及Ｂ７－２表达水平及ＤＣ诱导Ｔ细胞增殖能力。结果ＧＭ－ＣＳＦ及ＩＬ－４联合刺激选择性使ＰＢＭＣ中ＤＣ大量增殖,并通过增强ＤＣ表面ＨＬＡ－ＤＲ及Ｂ７表达［从（１２．８±１．１）、（１５．１±１．０）增至１９．１±１．７）、（２１．６±１．５）,Ｐ＜０．０１］进一步增强ＤＣ免疫功能［由（６８２０±１４０）增至（１４０９０±１８０）ｍｉｎ－１,Ｐ＜０．０１］。结论联合应用ＧＭ－ＣＳＦ及ＩＬ－４能够从肝癌患者血中制备出大量高免疫活力ＤＣ。     

膀胱癌可溶性抗原诱导细胞毒T细胞的实验研究

目的 观察膀胱癌可溶性抗原诱导细胞毒Ｔ细胞生长,为构建膀胱癌疫苗及过继性免疫治疗提供实验基础。方法 提膀胱癌可溶性抗原（ＴＳＡ）,用ＴＳＡ、抗ＣＤ－３单抗、ＩＬ－２联合诱导外周血单个核细胞（ＰＢＭＣ）,动态观察细胞增殖,进行细胞表型分析和细胞因子测定;并与ＩＬ－２诱导的ＬＡＫ细胞,抗ＣＤ－３单抗、ＩＬ－２诱导的ＣＤ３－ＡＫ细胞进行比较。结果 实验组细胞第８ｄ增殖７．５２倍,第１２ｄ增殖２８．９２倍     

雷公藤多甙和IL—10对人DC内IL—12p40和DC—CK1mRNA转录的影响

目的：研究雷公藤多甙和ＩＬ－１０对ＤＣ内ＩＬ－１２ｐ４０和ＤＣ－ＣＫ１转录的影响。方法：通过ＧＭ－ＣＳＦ、ＩＬ－４和ＴＮＦα体外培养体系,从人外周血单个核细胞中诱导ＤＣ,ＩＬ－１２０ｐ４０和ＤＣ衍生的趋化因子１（ＤＣ－ＣＫ１）转录采用逆转录多聚酶链反应（ＲＴ－ＰＣＲ）技术。结果：通过ＧＭ－ＣＳＦ、ＩＬ－４和ＴＮＦα体培养体系可以获得成熟功能性ＤＣ,雷公藤多甙和ＩＬ－１０能抑制ＤＣ内ＩＬ－１２ｐ４０     

雷公藤多甙对体外培养正常人外周血单个核细胞产生IL-5的抑制作用研究

目的：探讨雷公藤多甙（ＴⅡ）对体外培养正常人外周血单个核细胞（ＰＢＭＣ）产生ＩＬ－５的抑制作用，为其临床应用提供依据。方法：体外分离培养正常人ＰＢＭＣｓ，用ＥＬＩＳＡ法检测在不同ＴⅡ浓度干预下培养体系上清液中ＩＬ－５的浓度。结果：ＴⅡ在小于５ｍｇ·Ｌ１时对ＩＬ－５的产生即具有抑制作用，抑制作用与ＴⅡ的浓度呈明显剂量依赖性。在０．６２５ｍｇ·Ｌ１时可使ＩＬ－５的产生减少１２．１％，在５ｍｇ·Ｌ１时可使ＩＬ－５的产生减少７６．８％，与１０－６ｍｏｌ·Ｌ１的地塞米松相似。结论：ＴⅡ在无细胞毒作用的浓度下，能显著抑制体外培养ＰＢＭＣｓ产生ＩＬ－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.