目的:研究丹參酮ⅡA(Tan ⅡA)對急性早幼粒細胞白血病(APL)細胞株NB4細胞誘導的血管內皮細胞株(ECV304)促凝活性(PCA)的影響,并對其機制作初步探討。方法:(1)分別用1.0μg/mL TanⅡA、0.3μg/mLATRA、0.01%DMSO、PRMI1640處理NB4細胞24、48和72h,取其上清液作為條件培養基(hNB4-CM)。將這些CM分別與ECV304細胞在37oC共同孵育0、4、8和12h,用反復凍融法制備ECV304細胞裂解液,采用一期凝血法測定其PCA;采用ELISA法測定條件培養基中的TNF-α 。(2)ECV304細胞與1.0μg/mL TanⅡA及TanⅡA 72h-NB4-CM 在37oC共同分別孵育6、12、24和48h,并以ATRA和DMSO分別作為陽性和陰性對照,用上述相同方法測定ECV304細胞裂解液的PCA。結果:(1)1.0 μg/mL Tan ⅡA可以誘導NB4細胞分化,其作用NB4細胞的培養基有一定的升高ECV304細胞PCA的作用,該作用在孵育4h時達高峰,之后ECV304細胞PCA逐漸下降。與0.3μg/mL ATRA的作用無統計學差異(Pgt;0.05)。(2)1.0 μg/mL的TanⅡA對TanⅡA72h-NB4-CM促ECV304細胞PCA有抑制作用,其強度隨作用時間增加而增加,與1.0μmol/L ATRA比較,Pgt;0.05。(3)TanⅡA作用NB4細胞的培養基中TNF-α濃度,在作用前7h內隨作用時間增加而增加,與0.3μg/mL ATRA比較無差異(Pgt;0.05)。結論:Tan ⅡA能誘導NB4細胞分化,后者在分化過程中釋放的TNF-α可能與ECV304細胞PCA活性升高有關;Tan-ⅡA又能抑制Tan-ⅡA-NB4-CM增強ECV304細胞PCA的作用。
【摘要】 目的 觀察t(4; 22)致血小板源性生長因子受體α（the platelet-derived growth factor receptor alpha， PDGFRA）異常的髓系/淋巴系腫瘤的臨床特點。 方法 對2010年6月收治的1例t(4; 22)致PDGFRA異常的髓系/淋巴系腫瘤患者的臨床資料進行回顧性分析，并對其臨床特點、實驗室檢查、診斷、治療進行總結。 結果 該疾病臨床表現及骨髓涂片檢查類似慢性粒細胞白血病（chronic myelogenous leukemia，CML），但無CML特征性Ph染色體和（或）BCR/ABL融合基因，而細胞遺傳學檢測顯示4號與22號染色體易位，診斷為t(4; 22)致PDGFRA異常的髓系/淋巴系腫瘤。采用羥基脲及干擾素治療后可獲得完全血液學緩解。 結論 t(4; 22)致PDGFRA異常的髓系/淋巴系腫瘤是一類罕見疾病，臨床表現與CML相似，t(4; 22)及BCR/PDGFRA融合基因陽性是診斷該類疾病的關鍵。【Abstract】 Objective To observe the clinical features of myeloid and lymphoid neoplasms with t (4; 22) induced abnormalities of the platelet-derived growth factor receptor alpha (PDGFRA) to increase the identification and reduce the misdiagnosis. Methods The clinical data of one patient with myeloid and lymphoid neoplasm with t (4; 22) induced abnormalities of PDGFRA diagnosed in June 2010 was retrospectively analyzed. We summarized the clinical features, morphology, genetics, diagnostic criteria and therapy about this kind of disease. Results The patient had a clinical manifestation and bone marrow smear result of chronic myelogenous leukemia (CML). But the result of genetic analysis found no translocation of chromosomes 9 and 22 juxtaposing BCR and ABL gens. Cytogenetic analysis showed an abnormal karyotype with rearrangement of chromosomes 4 and 22. So the patient was diagnosed myeloid and lymphoid neoplasms with t (4; 22) induced abnormalities of PDGFRA. After receiving interferon and hydroxyurea, the patient achieved complete hematologic remission. Conclusion Myeloid and lymphoid neoplasms with t (4; 22) induced abnormalities of PDGFRA is a rare kind of disease. Its clinical feature is similar to that of CML. The key of diagnosis is genetics.
【摘要】 目的 分析異基因造血干細胞移植術（allogeneic hematopoietic stem cell transplantation，allo-HSCT）后出血性膀胱炎（hemorrhagic cystitis，HC）相關的危險因素，動態監測受者尿BK病毒（BK virus，BKV），分析其與HC發病的關系。 方法 回顧性分析2003年3月-2008年1月期間接受allo-HSCT的121例患者的資料，選擇8個臨床參數[年齡、性別、疾病類型、移植時疾病狀態、供者類型、預處理方案、急性移植物抗宿主病（acute graft-versus-host disease，aGVHD）、aGVHD的預防方案]作COX回歸分析。采用SYBR Green染料實時熒光定量聚合酶鏈反應法對2006年9月-2008年1月42例allo-HSCT患者尿BKV載量進行動態監測，分析被檢查者尿液BKV基因載量與HC發生以及嚴重程度的關系。 結果 121例患者中有24例發生HC，發病時間為術后0~63 d，中位時間40 d；持續時間7~150 d，中位時間22 d。Ⅱ~Ⅳ度aGVHD為HC的獨立危險因素[RR=8.304，95%CI（1.223，56.396），P=0.030]。allo-HSCT受者尿液中BKV檢出率為100%（42/42）。與正常人及未發生HC的allo-HSCT受者相比，HC患者尿中BKV基因載量具有更高平均峰值。 結論 Ⅱ~Ⅳ度aGVHD，尿中BKV DNA高載量與HC的發生有相關性。【Abstract】 Objective To identify the risk factors for hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (allo-HSCT), and define the quantitative relationship between BK virus (BKV) DNA load with HC. Methods The medical records of 121 patients undergoing allo-HSCT from March 2003 to January 2008 were retrospectively analyzed. Eight clinical parameters were selected for COX regression analysis, including age, sex, underlying disease, disease status at transplant, donor type, conditioning regimen, acute graft-versus-host disease (aGVHD), and GVHD prophylaxis. From September 2006 to January 2008, mid-stream urine samples were continuously collected from 42 patients with allo-HSCT. SYBR green real-time polymerase chain reaction, technique was utilized to define the quantitative relationship between BKV DNA load and HC. Results Twenty-four out of 121 patients developed HC. The median time of onset was 40 days after HSCT, ranged from 0 to 63 days. The disease lasted for 7 to 150 days, with a median duration of 22 days. Grade Ⅱ-Ⅳ aGVHD [RR=8.304, 95% CI (1.223,56.396); P=0.030] was identified as an independent risk factor for the occurrence of HC. BKV excretion was detected in 100% (42/42) of the recipients of allo-HSCT. When compared with asymptomatic patients and allo-HSCT recipients without HC, patients with HC had a significantly higher mean peak BKV DNA load. Conclusions Patients are at an increased risk of developing HC if they have grade Ⅱ-Ⅳ aGVHD. A correlation between the load of BKV and incidence of HC may exist.
目的 建立急性白血病（AL）患者八色流式免疫表型分析起始管方案。 方法 用胞膜CD3（CD3）、CD19、CD10、CD34、CD45、胞漿CD79a（cCD79a）、髓過氧化物酶（MPO)和胞漿CD3（cCD3）等8種抗體建立八色流式染色方案。膜表面抗體直接染色；膜內抗體經固定破膜，再染色后上機檢測。將3個血小板減少患者骨髓標本分別進行抗體的單色染色和缺一色染色；最后對17例確診的AL初發患者標本進行檢測。 結果 用單色染色來確定染色方案中各抗體的檢測電壓及熒光補償；缺一色染色中，陽性細胞群較單色染色變化均＜10%，表明方案中的各抗體相互作用小。17例AL初發患者中，6例急性B淋巴細胞白血病原始細胞均為CD34和CD19陽性，5例cCD79a陽性和4例CD10陽性；4例急性T淋巴細胞白血病患者均為cCD3陽性；6例急性髓細胞白血病均為CD34和MPO陽性；1例B+T混合表型AL患者CD34、cCD3、CD19、cCD79a及CD10均為陽性，MPO和CD3為陰性，此檢測方案能夠確定各類AL的細胞類型。 結論 建立了AL患者八色流式免疫表型分析起始管方案，操作簡便快速，適用于臨床檢測。
ObjectiveCD44 and CD54 are two specific biomarkers of gastric cancer stem cells and were used as targets in this study. The number of CD45–CD44+CD54+ cell subsets in peripheral blood of gastric cancer patients was detected by flow cytometry. Further, we combined these results with the clinicopathological characteristics of gastric cancer patients to analyze the significance of CD45–CD44+CD54+ cell subsets.MethodsFrom December 2016 to September 2017, 38 patients with gastric cancer in gastrointestinal surgery of West China Hospital of Sichuan University were included as the study object. The content of CD45–CD44+CD54+ cell subsets in their peripheral blood was detected by flow cytometry and its clinical significance was analyzed.ResultsThe median number of CD45–CD44+CD54+ cells were 541.9/mL (71.7–8 057.0/mL) in 38 patients and 555.9/mL (71.7–8 057.0/mL) in the group of patients with R0 resection. Patients without lymph node metastasis were found to have more CD45–CD44+CD54+ cells than patients with lymph node metastasis [941.4/mL (183.5–8 057.0)/mL vs 379.3/mL (71.7–2 269.7/mL, P=0.002], and more CD45–CD44+CD54+ cells in patients with TNM stage Ⅰ–Ⅱ than in TNM stage Ⅲ–Ⅳ [858.6/mL (183.5–8 057.0/mL) vs 364.6/mL (71.7–2 269.7/mL, P=0.015]. The patients with T3–4 stages (P= 0.025), N+ stage (P=0.009) and TNM Ⅲ–Ⅳ stage (P=0.012) had low ratios of the subgroup with high number of CD45–CD44+CD54+ cells, respectively. We made a more accurate judgment of N stage and TNM stage when we combined tumor size and the number of CD45–CD44+CD54+ cells together. However, there was no significant correlation between the number of CD45–CD44+CD54+ cells and other clinicopathological features and prognosis.ConclusionsThe number of CD45–CD44+CD54+ cell subsets is correlated with tumor progression, which might be used to predict TNM stage and N stage. However, the number of patients included in this study is too small, and the clinical significance of CD45–CD44+CD54+ subsets in gastric cancer patients needs to be further demonstrated by expanding the sample size.