Significance of cytogenetic and fluorescence in situ hybridization analysis in evaluating antichronic myeloid leukemia efficacy of different immune effector cells

We report on the antileukemia effect of interleukin 2 (IL2) on different immune cells from 22 patients with chronic myeloid leukemia (CML). Bone marrow cells from these patients were first cultured in modified long-term bone marrow culture medium for several days, then separately cultured with lymphokine activated killer cells (LAK), cytokine-induced killer cells (CIK), and dendritic cell cocultured CIK (DC-CIK) for another 1-2 days. They were then detected for presence of the Philadelphia chromosome (Ph) by cytogenetic analysis and fluorescence in situ hybridization (FISH). The percentage of Ph-chromosome-positive cells in the bone marrow mononuclear cells after culturing with CIK and DC-CIK was significantly lower than that after culturing with IL2 or LAK. Our results demonstrate that cytogenetics and FISH are useful techniques for the evaluation of the anti-CML effect of immune cells and that CIK or DC-CIK can be appropriate candidates for adoptive immune cell therapy in vivo or for leukemia cell purging ex vivo.     

经舟骨月骨周围脱位的诊断和治疗

目的 探讨经舟骨月骨周围脱位的诊断和治疗。方法 回顾性分析17例经舟骨月骨周围脱位病例，背侧型13例，掌侧型4例；新鲜损伤组10例中8例行撬拨复位成功，仅2例因舟骨对位不良接受手术治疗；陈旧性组7例中均接受手术，4例行开放复位内固定，3例行近排腕骨切除术。结果 采用Cooney评价标准，优11例，良3例，可1例，差2例，优良率82．35％。结论 经舟骨月骨周围脱位的早期治疗简单且疗效较好，而一旦误诊，则多数需手术治疗，且残留不同程度的功能障碍。提高对本病损伤机制、腕部体征和影像学特点的认识和了解，减少误诊。是提高本病预后的关键因素之一。     

肝脏特异性表达载体Kbpala/Alb-ATP7B 的构建及表达

目的：构建小鼠白蛋白启动子引导下携带野 生及常见突变型Arg778Leu、His1069Gln的人ATP7B cDNA的肝脏特异 性表达载体Kbpala/Alb-ATP7B，并检测其表达情况。方法:定点突变法获取人群中常见的Arg778Leu及His1069Gln两种ATP7B突变体，定向克隆将小鼠白蛋白启动子Alb序列及野生和突变的ATP7B cDNA依次亚克隆至转基因载体Kbpala上，得到可在肝脏特异性表达的正常及突变型人 ATP7B的转基因载体Kbpala/Alb-ATP7B。将其 短暂转染BRL细胞及BHK细胞，通过Western blotting检测其蛋白表达情况。结果:经酶切鉴定及定点测序证实，转基因载体Kbpala/Alb-ATP7B 构建成功；Western blotting显示仅在肝脏细胞实现表达。结论:带 有人类ATP7B cDNA的野生及常见致病突变型的转基因载体Kbpala/Alb -ATP7B构建成功并在肝脏细胞特异性表达。     

路氏乳杆菌JCM1081黏附相关蛋白的初步鉴定

目的研究路氏乳杆菌(Lactobacillus reuteri,也称罗伊氏乳杆菌)JCM1081菌体表面蛋白对其黏附HT-29细胞的影响。方法将路氏乳杆菌JCM1081菌体进行胰蛋白酶、蛋白酶K处理;用氯化锂和盐酸胍对乳杆菌表面的蛋白进行抽提,进行SDS-PAGE后与黏蛋白受体进行Western blot,并对杂交阳性蛋白进行质谱分析鉴定。结果路氏乳杆菌JCM1081菌体经胰蛋白酶、蛋白酶K处理后,其对HT-29细胞的黏附力显著下降(P<0．01);用氯化锂去除路氏乳杆菌JCM1081菌体外表面的S层蛋白后,路氏乳杆菌JCM1081对HT-29细胞的黏附力无显著变化;Western blot结果显示相对分子质量(Mr)为29×103和14×103的两种菌体表面蛋白与黏蛋白受体杂交中出现了强阳性;质谱分析结果显示29×103蛋白与路氏乳杆菌ATCC55730的h0793蛋白相似性高达71．1％。结论路氏乳杆菌JCM1081菌体表面的蛋白参与了乳杆菌的黏附,其中29×103和14×103的两种胞壁表面蛋白能够特异地识别黏蛋白受体并与之结合,29×103蛋白属ABC转运蛋白家族。     

低营养及低氧状态下NIH3T3细胞增殖及细胞周期的变化及对bFGF的影响

目的： 体外模拟慢性创面缺氧、低营养环境，观察成纤维细胞在该状态下增殖及细胞周期的变化及对外源性生长因子（bFGF）的反应,探讨低氧、低营养条件下成纤维细胞的病理生理变化。方法: 单纯缺氧环境采用厌氧培养箱，通入混合气，氧分压（PO₂）分为27 mmHg和44 mmHg 2个水平；低营养环境则控制培养液新生牛血清（NCS）浓度。用MTT法检测细胞活性以及其对外源性生长因子的反应，用流式细胞仪检测细胞周期。结果: PO₂ 44 mmHg时细胞增殖速度较同期对照组无明显差异；PO₂ 27 mmHg时，细胞增殖速度较同期对照组明显减慢（P<0.01），细胞被阻滞于G₀期，S期细胞比例明显减少，bFGF未显示促增殖作用。NCS浓度为0.5%的低营养状态下细胞增殖速度较同期对照组明显减慢（P<0.01），细胞被阻滞于G₀-G₁期（P<0.01）；bFGF能明显改善低营养状态下的增殖减慢（P<0.01），使G₂-M期细胞比例增加（P<0.05）。结论: 27 mmHg PO₂或NCS浓度为0.5%的低营养环境使细胞阻滞于G₀-G₁期，影响成纤维细胞增殖；bFGF可以改善低营养条件下细胞增殖减慢的状态，但对极度缺氧条件下的成纤维细胞增殖障碍无明显作用。     

用Millicell底膜培养皿研究Tourmaline对ECV-304细胞增殖的影响

目的 用联合培养的方法研究Tourmaline对体外培养的人血管内皮细胞(ECV-304)增殖作用的影响。方法 用放置Tourmaline微球的Millicell底膜培养皿与人血管内皮细胞ECV304联合培养。通过细胞形态、细胞融合面积、细胞计数及增殖细胞核抗原(PCNA)的免疫细胞化学等方法观察培养细胞增殖的变化。结果 联合培养24h后Tourmaline组(与对照组比)细胞形态正常,呈铺路石样生长,贴壁细胞覆盖率从(79.50±5.92)％长到(90.17±3.49)％(P<0.05),细胞数增加(16.62±4.14)％(P<0.05),PCNA阳性细胞为(43.50±3.19)％,与对照组比,差异均有非常显著意义(P<0.01)。结论 Tourmaline可通过加强DNA合成,促进体外培养的ECV-304细胞增殖。     

Deletion of stem-loop 3 is compensated by second-site mutations within the Gag protein of human immunodeficiency virus type 1

Encapsidation of human immunodeficiency virus type 1 (HIV-1) RNA involves specific interactions between viral Gag proteins and viral RNA elements located at the 5' untranslated region (UTR). These RNA elements are termed packaging (psi) or encapsidation (E) signals and mainly comprise the stem-loop 1 (SL1) and SL3 RNA structures. We have previously shown that deletion of the SL1 sequences is compensated by second-site mutations within Gag. Similar studies are now extended to SL3 and the results demonstrate that deletion of this RNA structure is rescued by two point mutations, i.e., A11V in p2 and I12V in nucleocapsid (NC). These two compensatory mutations are different from those associated with the rescue of SL1 deletion, suggesting that SL1 and SL3 may bind to different residues of Gag during viral RNA packaging. Analysis of virion-derived RNA in native agarose gels shows that deletion of SL3 leads to decreases in both viral RNA packaging and dimerization. These defects are corrected by the compensatory mutations A11V and I12V. Yet, defects in viral RNA dimerization at an early stage that were caused by the SL3 deletion in the context of a viral protease-negative mutation cannot be overcome by these two suppressor mutations. Therefore, the positive effects of A11V and I12V on dimerization of the SL3-deleted RNA must have taken place at the maturation stage.     

合成消旋丹参素及其衍生物对血小板聚集性和cAMP含量的影响

合成消旋丹参素(简称合成丹参素)及其衍生物(BLA、BLE)在体外实验和体内静脉注射时,对ADP诱导的家兔和大鼠血小板聚集性均有明显的抑制作用。BLK和BLE有较强的促进血小板解聚作用,而合成丹参素较弱。合成丹参索、BLA和BLE在体外实验时,无升高血小板内cAMP作用,反使其cAMP含量下降。提示合成丹参素及其衍生物对血小板功能的作用,不是通过提高cAMP水平;推测可能和抑制血栓素A_2(TXA_2)合成酶有关。