Preparation and clinical application of immuno-magnetic latex

在最近十年內，周邊造血幹細胞的移 植，廣泛地應用於癌症的治療。異體移植 可以用來解決不同的的血液疾病，自體移 植對於高劑量化療的病患，可以加速病患 骨髓的回復，防止感染或出血的併發症發 生。但是自體移植總有癌細胞污染之疑 慮，而異體移植亦有移植物抗宿主病的難 題。所以，選取CD34+細胞作為移植可能 可以避免上述兩項移植的缺點而達到更好 的效果。在一般商業化選取CD34+細胞是 利用磁場分離(magnetic cells sorting)。但是 MACS 所使用的帶有anti-CD34 單株抗體 的免疫磁性乳膠顆粒是非常貴的。每處理 一次約需20 萬台幣。我們嘗試利用我們自 製的免疫磁性乳膠顆粒上的COOH 官能基 鍵結anti-CD34 單株抗體，去做相同純化的 療程。我們相信自製的免疫磁性乳膠顆粒 會較便宜，病人也能夠負擔得起。我們希 望此技術能夠進行，而且期待可以推廣至 其他負向選取（negative selection），例如接 枝anti-CD4 或anti-CD8 去移除CD4+ 細 胞或CD8+細胞，用來治療一些自體免疫方 面的疾病。Peripheral blood stem cell transplantation (PBSCT) is widely used in this decade for cancer treatment. Allogeneic setting can be used for various blood diseases and autologous setting may be benefit for solid tumors as a rescue after high-dose chemotherapy to accelerate the bone marrow recovery and prevent the infections or bleeding complications. But, possibility of tumor cells contamination is a pitfall of autologous PBSCT while increased incidence of graft vs. host disease may decrease the successful rate in allogeneic PBSCT. So, CD34+ cells selection will solve the above problems. The commonly used and commercially available method of CD34+ cells selection is magnetic cells sorting. But the beads with anti-CD34+ monoclonal antibodies are rather expensive. It needs 200,000 NT per procedure. So, We try to bind anti-CD34 to our homemade magnetic beads using –COOH bounding in order to do the same purging procedure. We do believe the homemade price will be rather cheap and can be afforded by patients themselves. We hope this technique will work and it can be expanded to other negative selection such as anti-CD34 or anti-CD8 to remove CD4+ or CD8+ cells in order to treat some antoimmune diseases

高分子薄膜於外加電場下應用於血漿蛋白質分離的研究

過去實驗已證實自製之高分子薄膜於 分離免疫球蛋白及白蛋白之可行性，惟其 間發現分離之效果未臻理想之最大原因為 蛋白質在薄膜表面形成一層濾餅（cake）， 為改進此缺失，本實驗乃以掃流過濾及外 加電場，以期藉流動性溶液及電場使免疫 球蛋白不致沉積在薄膜表面，阻塞過濾效 果，以便白蛋白通過薄膜，如此將可達到 預期的效果。故本實驗分為三部分：（1） 薄膜結構之鑑定包括薄膜之表面，截面， 孔隙度大小之測定（2）蛋白質溶液在外加 電場掃流之過濾實驗及（3）血液檢體於同 樣狀況下之測試。希望此外加電場掃流之 簡易裝置能徹底改善過去因阻塞造成阻力 增加，導致過濾效果降低之缺失，使此高 分子薄膜能作為臨床上治療性血漿分離時 二次過濾之用。Previous experiments have proved that membranes have the possibility to separate of globulins and albumin. From these experiments, we found the pitfall causing the lower filtration efficiency is due to the “cake” formation on the membranes. In order to improve this pitfall, we try to add an electric field and cross-flow to the experiments. As we know, the anode will attract the negativechanged globulins with the cross-flow that will prevent the obstruction of the micropores. By the way, the cross-flow will simulate the condition of cascade filtration by its continuous flow. So, three parts will compose the studies: 1) Identification of membrane structures including surface and cross-section, measuring size of the micropores, contact angles and degree of crystallization. 2) Experiments of protein solutions filtered through membranes within a cross-flow device under electric field. 3) Experiments of plasma of different composition filtered through membranes under the same conditions of above. We hope this simple cross-flow and electric field design will improve the filtration efficacy and let the membrane can be used clinically in cascade or double filtration technique in therapeutic plasmapheresis

不同薄膜之表面特性與生物適應性之影響

Monocyte adhesion and subsequent activation are major events that facilitate the foreign-body reaction. These studies evaluate the effect of semicrystalline polyamide (Nylon-66), poly (ethylene-co-vinyl alcohol) (EVAL), and poly (vinylidene fluoride) (PVDF) with nonporous and porous morphologies on the ability of monocyte adhesion and activation to produce variable levels of IL-1b, IL-6, and TNF-a. Results indicated IL-1b was produced in the greatest quantity by these polymers. In addition, monocyte adhesion and activation on a material may alter to a great extent dependent on the surface morphology and wettability. As the membrane wettability increases, the ability of the membrane to adhere monocytes increases but to stimulate monocyte production of cytokines decreases. Similarly, these membranes when prepared with porous surfaces can enhance monocyte adhesion and suppress monocyte activity. Therefore, the nonporous PVDF membrane is the least biocompatible in this work. In contrast, the hydrophilic membrane Nylon-66 with porous surface is the least stimulating of monocyte cytokine production when compared to all of the other membranes evaluated with nonporous or porous surface. These studies provide important insight into conditions that modulate monocyte activity in response to the substratum morphology and wettability

DFO經皮給藥的可行性研究

海洋性貧血病人需長期接受輸血以維持生命，然而長期輸血容易導致病人體 內三價鐵離子含量過高而產生鐵中毒的現象，鐵中毒慣例上採用deferoxamine mesylate(Desferal, DFO)連續式點滴或皮下注射以螯合治療，但病患容易因為疼痛 而產生排斥感，使治療效果不佳，所以我們希望能夠改變藥物劑型，企圖以經皮 給藥的方式來取代注射治療，本實驗的目的即是評估將DFO 製成貼劑貼在皮膚上 給藥的可行性，將選擇基質式經皮吸收系統，使DFO 能穿過皮膚，藉以設計DFO 貼片。此貼片希望能夠以一定傳送速率讓藥穿過皮膚到達循環系統，而達全身性 治療效果。本實驗係以商用感壓膠為基質，探討DFO 在其中的藥物釋放情形，以 開發為經皮吸收貼劑。The goal of this study is to help thalassemia patients, who chronically require blood transfusions to keep alive. However, such long-term transfusions can easily lead to a build-up of trivalent iron and produce an iron overload condition. To treat iron overload, the patients usually receive intravenous or subcutaneous deferoxamine mesylate (DFO) regularly. But, because of the pain the patients may resist taking drug, so the effects of treatment are not very good. Consequently, we hope to be able to change the method of delivering DFO—we want to see if we can replace the injection method with transdermal drug delivery. The preliminary goal of the study, therefore, is to assess the feasibility of designing and fabricating a DFO patch using an adhesive dispersion-type transdermal drug delivery system. It is hoped that it can let the drug pass through the skin at a fixed rate of speed and reach the circulation system, from where it can spread its therapeutic effect throughout the body. In this research, we choose the commercial pressure sensitive adhesives to be the drug reservoirs and observe the release profile of DFO in the adhesives in order to develop and design transdermal patchs

唐氏症兒童的血液性疾病：臺大醫院的經驗

Background and Purpose: Children suffering from Down's syndrome (DS) are predisposed to the development of neoplastic disorders of the hematopoietic system and tend to display many unique characteristics. This study analyzed the clinical characteristics and treatment outcomes of 16 children with DS who developed neoplastic disorders. Methods : All DS patients aged <18 years of age with a diagnosis of leukemia or myelodysplastic syndrome (MDS) from 1990 to 2002 were included in this retrospective study. The clinical and laboratory characteristics of patients, including age at diagnosis, gender, initial hemogram, cytogenetic findings, immunophenotype, treatment regimen and outcomes were analyzed. Results: Sixteen patients met the inclusion criteria. Among them, 9 patients (56%) had acute myeloid leukemia (AML), mainly of the megakaryoblastic subtype. All 8 AML patients who had analyzable metaphase cells revealed clonal chromosomal abnormalities in addition to trisomy 2l. Three of these patients developed MDS prior to the onset of AML. Of the 5 patients who underwent chemotherapy, 8 remained in remission with a survival time of 29, 59, and 109 months, and the remaining 2 died as a consequence of chemotherapy toxicity. Among the 6 patients (38%) who developed transient myeloproliferative disorder, 2 were lost to follow-up, 2 died from DS-associated congenital heart abnormalities and 2 survived without any AML changes. The remaining 1 patient (6%) who developed ALL was still in his first remission although this patient suffered profound chemotherapy complications during treatment. Conc1usions. This study found that AML is the most common hematologic neoplasm in Taiwanese children with DS, especially megakaryoblastic leukemia. This finding is comparable to the reported results from related studies in different countries. Long- term remission of AML in DS patients can be achieved with appropriate treatment