多点实时剂量监测在全身照射中的应用

  目的  探讨在有限条件下完成分次双侧位大野全身照射治疗时多点实时剂量监测技术用于质控的必要性。  方法  使用Varian 2300 C/D加速器6 MV射线对25例准备进行骨髓移植的患者进行全身照射治疗。治疗同时使用半导体剂量计进行多点实时剂量监测, 记录监测结果并与计划计算结果进行比对分析, 在治疗的同时进行剂量的修正, 以确保患者受照剂量误差控制在±5%以内。  结果  25例患者的12通道首次实时剂量监测结果与理论计划剂量有显著偏差。头部、胸部、腹部、盆腔、大腿、膝关节和踝关节剂量偏差中位数分别为6.10%、-1.30%、5.90%、5.05%、-4.00%、3.05%和3.30%。通过对实时监测数据的分析及时修正偏差, 所有患者的受照剂量偏差均在±5%以内, 上述部位的剂量偏差中位数分别降为0.35%、-1.05%、0.60%、0.20%、-2.55%、1.20%和0.85%。  结论  实时剂量监测是实施全身照射的必要质控手段。     

骨髓干细胞移植治疗骨不连及骨缺损临床研究

目的观察自体骨髓干细胞移植治疗骨不连及骨缺损的疗效。方法140例患者随机分为自体髂骨移植组(A组)及自体骨髓干细胞移植组(B组)各70例,均行手术治疗,并行简单有效的内外固定。结果术后平均愈合时间A组为(7.0±2.0)个月,B组为(5.0±1.5)个月,两组间有显著性差异(P<0.05);治疗期间患者无明显不良反应。结论自体骨髓干细胞移植较传统植骨治疗骨不连及骨缺损疗程短、疗效好,无不良反应。     

人骨髓间充质干细胞输注对辐射损伤小鼠造血器官的保护作用

本研究探讨人骨髓间充质干细胞(MSC)对辐射损伤小鼠造血器官的保护作用。培养人骨髓MSC,并进行细胞学鉴定。BALB/c雌性小鼠经6 Gy60Coγ射线照射后,分别通过尾静脉注射不同剂量的人MSC、MSC裂解物或等体积生理盐水。每天记录小鼠存活情况,并于处理后的第9天和16天取小鼠股骨和脾脏进行病理学分析,观察造血组织容量,并进行细胞增生程度评分。结果表明,与对照组比较,MSC及细胞裂解物处理组小鼠生存期无明显延长,但第9天高剂量(5×106)MSC及裂解物组小鼠骨髓造血即部分恢复,造血组织容量显著改善,(43.50±12.08)%和(42.80±13.11)%vs(24.33±9.50)%(P<0.05),增生程度积分明显增高(P<0.05),脾脏出现增生结节。第16天高剂量MSC和裂解物处理组小鼠骨髓和脾脏细胞明显活跃,脾脏和骨髓结构接近正常小鼠。此外,两个时间点MSC与细胞裂解物处理组小鼠骨髓增生积分无明显差异。结论:骨髓MSC可促进辐射损伤小鼠的造血功能恢复,这种作用可能与细胞内固有的活性物质有关。     

骨髓内皮细胞条件培养液促进小鼠胚胎干细胞向造血分化

为观察骨髓内皮细胞条件培养液(BECM)和(或)细胞因子(VEGF SCF EPO)对小鼠胚胎干细胞(ESC-D3细胞)生成造血干／祖细胞的促进作用，先将ESC-D3细胞形成4天胚状体(Day 4 embryoid body,4dEB)，再诱导4dEBs生成造血干／祖细胞。实验分4组，第1，2和3组分别为BECM VEGF SCF EPO，BECM和VEGF SCF EPO组，第4组为自发分化对照组。检测各组造血干／祖细胞特异抗原、造血转录因子表达以及造血集落的形成。结果显示，BBCM和(或)细胞因子诱导生成的细胞均表达造血干／祖细胞抗原(c-kit,Sca-1,Thy-1和CD34)和造血转录因子(c—myb，SCL和β-H1)基因mRNA，培养后可产生HPP-CFC和BFU-E。从诱导ESC-D3细胞生成造血干／祖细胞的数量和生成的集落总数看，BBCM联合细胞因子组诱导效率均显高于单用组和对照组。结论：骨髓内皮细胞条件培养液能显促进胚胎干细胞早期造血分化，且骨髓内皮细胞条件培养液与细胞因子联合时效果更强。     

Stereotactic Body Radiation Therapy for Liver Cancer: A Review of the Technology

Stereotactic body radiation therapy has been adopted in the treatment of liver cancer because of its highly conformal dose distribution when compared with other conventional approaches, and many studies have been published to report the positive clinical outcome associated with this technique. To achieve the precision needed to maintain or to improve the therapeutic ratio, various strategies are applied in different components in the stereotactic body radiation therapy process. Immobilization devices are used in minimizing geometric uncertainty induced by treatment positioning and internal organ motion. Along with a better definition of target by the integration of multimodality imaging, planning target volume margin to compensate for the uncertainty can be reduced to minimize inclusion of normal tissue in the treatment volume. In addition, sparing of normal tissue from irradiation is improved by the use of high precision treatment delivery technologies such as intensity-modulated radiotherapy or volumetric modulated arc therapy. Target localization before treatment delivery with image guidance enables reproduction of the patient's geometry for delivering the planned dose. The application of these advanced technologies contributes to the evolution of the role of radiation therapy in the treatment of liver cancer, making it an important radical or palliative treatment modality.     

Single Fraction External Beam Radiation Therapy in the Treatment of Localized Metastatic Bone Pain. A Review.

Bone metastases are a frequent complication of cancer, and frequently cause pain. Indications for radiotherapy for bone metastases include pain, risk for pathologic fracture, and neurological complications arising from spinal cord compression, nerve root pain or cranial nerve involvement. There are numerous fractionation patterns of external beam radiation therapy for painful bone metastasis, both fractionated schedules and single fraction regimens. All prospective randomized trials that evaluated differences in the outcomes associated with various fractionated regimens versus single fraction regimens unequivocally showed that single fraction regimens (mostly 8 Gy) are at least equal with various fractionated regimens. The single fraction regimens have an additional advantage of being more convenient to both patients and hospitals. However, there are still numerous questions that are left unanswered in these trials, such as the “optimal” single fraction that should be used, the possibility for retreatment, and prognostic factors that may help identify those patients more likely to respond to a single fraction radiation therapy in the treatment of painful bone metastasis.     

骨髓间充质干细胞移植对豚鼠皮肤深Ⅱ度烧伤的治疗作用

目的探讨骨髓间充质干细胞 (MSCs)治疗皮肤深Ⅱ度烧伤的机制。方法体外培养扩增的MSCs悬液 ,以 2× 10 6/ml(A组 )和 2× 10 7/ml(B组 )两种细胞密度移植给皮肤深Ⅱ度烧的受体鼠创面 ,观察创面愈合速度 ,用免疫组织化学检测因子Ⅷ的表达。细胞移植后的第 15、3 0、5 0天 ,通过PCR方法检测创面皮肤组织中移植供体鼠的Y染色体基因表达。结果A、B组治疗侧的创面愈合速度均快于对照侧 ,A、B组治疗侧间无显著性差异。因子Ⅷ检测示第 15、3 0天时 ,A、B两组的微血管密度均多于对照侧 ,A、B组治疗侧间无显著性差异。PCR检测结果表明 ,第 15、3 0、5 0天时在部分受体鼠恢复的创面上有供体鼠Y染色体基因表达。结论骨髓MSCs对皮肤深Ⅱ度烧伤具有明显的疗效。     

骨髓间充质干细胞在心血管疾病中的研究及应用

背景：骨髓间充质干细胞具有多向分化潜能,对肝脏疾病、神经系统疾病、肺脏疾病、肾脏疾病的治疗都显示出其优越性,对心血管疾病治疗的研究也日益增多。目的：对骨髓间充质干细胞在心血管疾病中的应用以及治疗心血管疾病的机制进行综述。方法：应用计算机检索CNKI和Pubmed数据库中1985年1月至2009年12月关于骨髓间充质干细胞治疗心血管疾病的文章,在标题和摘要中以＂骨髓间充质干细胞,细胞移植,治疗,心血管疾病＂或＂bone marrow mesenchymal stem cells,cells transplant,therapy,cardiovascular disease＂为检索词进行检索,最终纳入42篇文献进行综述。结果与结论：众多的研究表明骨髓间充质干细胞可以在梗死的心肌中分化形成有功能活性的心肌、血管内皮等组织,改善心脏的功能,干细胞移植在治疗心血管疾病方面已表现出传统治疗方法所无可比拟的优越性。     

硼替佐米治疗初治多发性骨髓瘤成功病例报告(附文献复习)

目的:探讨蛋白酶体抑制剂——硼替佐米对初治多发性骨髓瘤的疗效及对移植造血干细胞采集的影响。方法:对一例初发的中年男性多发性骨髓瘤患者使用硼替佐米 地塞米松 反应停(VTD)的方案进行化疗,获得缓解后采集外周血造血干细胞。结果:应用以硼替佐米为基础的方案治疗3个疗程后,患者即获得完全缓解;完成4个疗程化疗后成功采集足够数量的外周血造血干细胞;完成6个疗程化疗后,进入维持治疗,至今已完全缓解17个月。治疗过程中除恶心、呕吐外无其他明显不良反应。结论:硼替佐米用于初治多发性骨髓瘤有良好的治疗效果,不良反应少,不影响造血干细胞采集。     

TBI和环磷酰胺选择性去除同种异基因反应供者淋巴细胞的研究

本研究用全身照射(TBI)和环磷酰胺(CY)选择性去除同种异基因反应的供者淋巴细胞,为预防移植物抗宿主病(GVHD)的发生探索新的手段。以(BALB/c×C57BL/6)F1雌性小鼠(H-2d/b)为受鼠,于第0天接受亚致死量的60Co-γ射线全身照射,总剂量为4Gy,第1天接种P388D1白血病细胞,第2天输注由C57BL/6雄性小鼠(H-2b)为供鼠提供的MHC不匹配的供者脾淋巴细胞,在造血干细胞移植前诱导移植物抗白血病效应(GVL)。第6天腹腔注射环磷酰胺(CY)200mg/kg或再次TBI9Gy,选择性去除同种异基因反应供者淋巴细胞,第7天输注(BALB/c×C57BL/6)F1雄性小鼠(H-2d/b)提供的骨髓造血干细胞。结果显示:以CY和TBI选择性去除同种异基因反应供者淋巴细胞组的小鼠无白血病和GVHD的发生,生存期超过了210天,于移植后第21天出现完全供者嵌合,然后嵌合率下降,第90天表现为混合嵌合体(MC)。对照组的小鼠出现了白血病和GVHD,出血、感染明显,生存期短,为20-36天(P<0.01)。结论:同基因骨髓移植前输注不相匹配的供者脾细胞诱导GVL效应,然后再通过TBI和CY选择性去除同种异基因反应的供者淋巴细胞来预防移植物抗宿主病(GVHD)的发生是可能的。     

Transplantation of autologous mononuclear bone marrow stem cells in patients with peripheral arterial disease (The TAM-PAD study)

Objectives For patients with severe, chronic limb ischemia in many cases interventional or surgical treatment is not possible anymore. In the past, both intramuscular and intraarterial transplantation of autologous BMCs had been proved therapeutically beneficial. The TAMPAD study is the first one to analyze combined intraarterial and intramuscular BMC transplantation in its acute and long-term effects. Methods 13 patients with chronically ischemic limbs due to peripheral arterial disease (Fontaine stage IIb) were recruited and underwent follow-up examinations after 2 and 13 months. Mononuclear cells from bone marrow were injected intramuscular and intraarterial into the ischemic limb. Results In contrast to the control group, after 2 months the pain-free walking distance of the transplanted patients significantly increased (from 147 ± 90 to 500 ± 614 m, p = 0.001). Furthermore the ankle-brachial index was significantly improved (at rest from 0.66±0.18 to 0.80±0.15, p=0.003, after stress from 0.64 ± 0.19 to 0.76 ± 0.16, p = 0.006). Similar improvement was documented in capillary-venous oxygen-saturation (thigh from 59 ± 9 to 66 ± 5, p = 0.005, lower leg from 56 ± 14 to 63 ± 5, p = 0.021) and venous occlusion plethysmography (rest from 2.1 ± 0.7 to 2.5 ± 0.7, p = 0.009, mean reactive hyperemia from 5.3 ± 1.8 to 7.2 ± 1.8, p = 0.003, and peak flow from 7.2 ± 3.2 to 10.8 ± 2.8, p = 0.002). After 13 months these positive effects persisted at their improved level. No side effects or complications were monitored. Conclusions Combined intraarterial and intramuscular transplantation of autologous mononuclear bone marrow stem cells is a clinically feasible and minimally invasive therapeutic option for patients with severe chronic peripheral occlusive arterial disease.     

基因修饰骨髓间充质干细胞修复关节软骨损伤

背景：随着生物技术的发展,通过转基因技术修饰细胞,从而获得长期稳定表达的生物活性因子以治疗关节软骨损伤逐渐引起重视。目的：就基因修饰的骨髓间充质干细胞在修复关节软骨损伤中的应用作一综述。方法：由第一作者检索1990至2011年PubMed数据库（http：／／www．ncbi．nlm．nih.gov／PubMed）有关基因修饰骨髓间充质干细胞修复关节软骨损伤的文献,英文检索词为“cartilage,genetherapy,mesenchymalstemcells,tissueengineering,bioactivefactor,vector”。共纳入15篇文献归纳总结。结果与结论：骨髓间充质干细胞已被广泛应用于修复关节软骨损伤。通过转基因技术将特定外源基因导入骨髓间充质干细胞,联合细胞治疗和基因治疗可达到更好的治疗效果,在关节软骨损伤的治疗中有广阔的应用前景。     

构建能转染人骨髓间充质干细胞的Ad5 GM-CSF-IL-2腺病毒载体

背景：如何将树突状细胞和T细胞活化因子运送至肿瘤局部是有待解决的难题之一。利用人骨髓问充质干细胞具有肿瘤趋向性和低免疫原性的特性,将树突状细胞和T细胞活化因子导入骨髓间充质干细胞后运送至肿瘤局部表达可能是解决上述难题的方案之一。目的：构建共表达粒细胞-巨噬细胞集落刺激因子和白细胞介素2的5型腺病毒载体（Ad5GM-CSF-IL-2）,感染人骨髓间充质干细胞,检测粒细胞一巨噬细胞集落刺激因子和白细胞介素2的表达水平和持续时间,为体内活化树突状细胞和T细胞提供实验依据。方法：提取人外周血单个核细胞总RNA,以此为模板用RT-PCR方法扩增粒细胞-巨噬细胞集落刺激因子和白细胞介素2基因cDNA,PCR扩增片段分别插入pcDNA3．1／Myc-His（-）B真核表达载体中,再利用脑心肌炎病毒内部核酸进入位点（IRES）序列,构建腺病毒载体穿梭质粒pDC515GM-CSF-IRES-IL-2。采用AdMax^TMFLP重组腺病毒载体体系,将穿梭质粒pDC515GM-CSF-IRES-IL-2与pBGHfrt△E1,3FLP骨架质粒共转染至293细胞,通过重组酶位点特异性重组获得Ad5GM-CSF-IL-2。Ad5GM-CSF-IL-2感染人骨髓间充质干细胞后经v射线照射使其失去增殖能力,分别用粒细胞-巨噬细胞集落刺激因子和白细胞介素2ELISA试剂盒在不同的时间点检测细胞上清中粒细胞-巨噬细胞集落刺激因子和白细胞介素2蛋白的水平。结果与结论：经测序证实,克隆获得的粒细胞-巨噬细胞集落刺激因子、白细胞介素2的cDNA序列分别与GenBank NM_000758（435bp）和GenBank NM_000586（462bp）提供的序列完全一致。用AdMax^TMFLP重组腺病毒载体可高效、快捷地获得所要包装的腺病毒载体,通过IRES连接粒细胞-巨噬细胞集落刺激因子和白细胞介素2两个基因,均获得高效表达。Ad5GM-CSF-IL-2感染人骨髓间充质干细胞后可连续7d高水平地分泌粒细胞-巨噬细胞集落刺激因子和白细胞介素2。     

Point‐of‐care treatment of focal cartilage defects with selected chondrogenic mesenchymal stromal cells—An in vitro proof‐of‐concept study

Due to the poor self‐healing capacities of cartilage, innovative approaches are a major clinical need. The use of in vitro expanded mesenchymal stromal cells (MSCs) in a 2‐stage approach is accompanied by cost‐, time‐, and personnel‐intensive good manufacturing practice production. A 1‐stage intraoperative procedure could overcome these drawbacks. The aim was to prove the feasibility of a point‐of‐care concept for the treatment of cartilage lesions using defined MSC subpopulations in a collagen hydrogel without prior MSC monolayer expansion. We tested 4 single marker candidates (MSCA‐1, W4A5, CD146, CD271) for their effectiveness of separating colony‐forming units of ovine MSCs via magnetic cell separation. The most promising surface marker with regard to the highest enrichment of colony‐forming cells was subsequently used to isolate a MSC subpopulation for the direct generation of a cartilage graft composed of a collagen type I hydrogel without the propagation of MSCs in monolayer. We observed that separation with CD271 sustained the highest enrichment of colony‐forming units. We then demonstrated the feasibility of generating a cartilage graft with an unsorted bone marrow mononuclear cell fraction and with a characterized CD271 positive MSC subpopulation without the need for a prior cell expansion. A reduced volume of 6.25% of the CD271 positive MSCs was needed to achieve the same results regarding chondrogenesis compared with the unseparated bone marrow mononuclear cell fraction, drastically reducing the number of nonrelevant cells. This study provides a proof‐of‐concept and reflects the potential of an intraoperative procedure for direct seeding of cartilage grafts with selected CD271 positive cells from bone marrow.     

Bone Marrow Edema Syndrome of the Medial Femoral Condyle Treated With Extracorporeal Shock Wave Therapy: A Clinical and MRI Retrospective Comparative Study

Objective

To determine the validity of extracorporeal shock wave therapy (ESWT) in the treatment of bone marrow edema (BME) of the medial condyle of the knee.

Mayo Alliance Prognostic Model for Myelodysplastic Syndromes: Integration of Genetic and Clinical Information

Objective

To develop a new risk model for primary myelodysplastic syndromes (MDS) that integrates information on mutations, karyotype, and clinical variables.

Clinical Applications and Utility of a Precision Medicine Approach for Patients With Unexplained Cytopenias

ObjectiveTo demonstrate experience and feasibility of a precision medicine approach for patients with unexplained cytopenias, defined as low blood counts in one or more cell lineages, persistent for 6 months or longer, in the absence of known nutritional, autoimmune, infectious, toxic, and neoplastic (secondary) causes.Patients and MethodsPatients were evaluated in our clinic between November 8, 2016, and January 12, 2018. After a thorough evaluation of known causes, family history, and appropriate clinical assays, genomic evaluation was performed in a stepwise manner, through Sanger, targeted, and/or whole-exome sequencing. Variants were analyzed and discussed in a genomics tumor board attended by clinicians, bioinformaticians, and molecular biologists.ResultsSixty-eight patients were evaluated in our clinic. After genomic interrogation, they were classified into inherited bone marrow failure syndromes (IBMFS) (n=24, 35%), cytopenias without a known clinical syndrome which included idiopathic and clonal cytopenias of undetermined significance (CCUS) (n=30, 44%), and patients who did not fit into the above two categories (“others,” n=14, 21%). A significant family history was found in only 17 (25%) patients (9 IBMFS, 2 CCUS, and 6 others), whereas gene variants were found in 43 (63%) patients (34 [79%] pathogenic including 12 IBMFS, 17 CCUS, and 5 others]. Genomic assessment resulted in a change in clinical management in 17 (25%) patients, as evidenced by changes in decisions with regards to therapeutic interventions (n=8, 47%), donor choice (n=6, 35%), and/or choice of conditioning regimen for hematopoietic stem cell transplantation (n=8, 47%).ConclusionWe show clinical utility of a real-world algorithmic precision medicine approach for unexplained cytopenias.