力学刺激对3月龄骨质疏松大鼠成骨细胞增殖与合成功能的影响

探讨周期性双轴力学应变对成骨细胞增殖与分化合成功能的影响。将正常 3月龄雌性 SD大鼠和骨质疏松大鼠颅顶骨分离的成骨细胞分别在含 10 %胎牛血清的 F- 12培养液中培养 ,并接种在双轴力学应变装置中。当细胞生长至亚融合状态 (Subconfluence) ,给细胞施加力学刺激 ,频率为 1Hz,力学刺激分别为 4 0 0、10 0 0、4 0 0 0μ strain;作用时间分别为每天 30 m in,2、4、8h,共加载两天。以未受力学刺激的细胞为对照组 ,受力学刺激的细胞为实验组 ,并进行比较。采用流式细胞技术测定细胞增殖变化 ;采用同位素标记方法检测成骨细胞骨钙素、I型胶原 C端前肽 (PICP)和总蛋白的分泌量。结果表明 :1)在静态培养条件下 ,3ovx组与 3control组比较 ,其细胞功能活性无明显变化 ,但 3ovx组大鼠成骨细胞增殖活性明显增高 ,这与绝经后骨质疏松骨代谢的高转换率相一致。 2 )4 0 0、10 0 0 μstrain力学刺激可以促进 3control组成骨细胞 I型胶原、骨钙素和总蛋白的分泌量增加 ,促进成骨细胞的分化成熟 ;在 10 0 0μstrain力学刺激下 ,成骨细胞合成骨基质的能力增加最为明显。同时 ,在 4 0 0、10 0 0μstrain力学刺激的初期也可以促进成骨细胞的增殖 ,而促进成骨细胞的分化成熟的作用大于促进细胞增殖的作用。 3)在4 0 0 0 μ     

阶跃切应力对内皮细胞释放前列环素的影响

采用流室定量研究阶跃切应力对内皮细胞释放前列环素（ｐＧＩ＿２）的影响。对流室中的内皮细胞连续１００分钟施加１、１０、２０ｄｙｎｅ／ｃｍ￣２的阶跃剪切应力，用放免法测流室的８个时间段流出液的６－ｋｅｔｏ－ＰＧＦｉ＿（１ａ）。绘出切应力刺激内皮细胞释放ＰＧＩ＿２的曲线。结果表明：未施加切应力时，内皮细胞释放ＰＧＩ＿２的量在检测限以下，施加切应力后，ＰＧＩ＿２释放速率很快达极大值，且又在数分钟内减小．施加阶跃切应力越大，ＰＧＩ＿２释放速率曲线峰值越高，但衰减也越快，在８０～１００分钟时间内的前列环素释放速率平均值也越大。由此提示，在活体中血液流变学因素一血流切应力对血管内皮细胞释放前列环素有促进作用。     

Kidney transplant monitoring by anti donor specific antibodies

Donor-specific anti-HLA antibodies were studied by cytotoxicity crossmatching (CTXM) and flow cytometry crossmatching (FCXM) in 117 kidney transplant candidates; the same study was carried out in 33 cadaver-donor kidney recipients, during the first 3 post-transplant months, for which donor cells were available. Pre-transport evaluation showed that 82.9 % of subjects were CTXM negative/FCXM negative, 6.8 % of patients were positive in both tests, and 10.3 % were CTXM negative/FCCM positive. Post-transplant monitoring for donor-specific antibodies (Abs-DS) showed that nine recipients (27.3 %) were FCXM positive; six of them were IgG + and three IgM +. In comparing these results with the clinical course, a significant association between FCXM IgG + and rejection episodes was observed (P < 0.01).     

肝硬化病人门静脉和肝动脉多谱勒血流参数变化的意义

目的 :探讨肝硬化门脉高压病人门静脉和肝动脉彩色多谱勒血流参数变化的意义。方法 :肝硬化伴食道静脉曲张的病人 6 6例 ,正常人对照组 5 8例。测量门静脉血流速度 (PVV)和肝动脉阻力参数 (HA- PI) ,计算肝血管指数。结果 :在肝硬化门脉高压病人比对照组 PVV明显降低 (12± 2 .2 ) cm/ s;(15 .8± 2 .6 ) cm/ s,P<0 .0 0 1)和 HA- PI明显增高(1.3± 0 .15 ;0 .94± 0 .16 ,P<0 .0 0 1) ,肝血管指数明显减低 [(8.6± 2 .1) cm/ s;(16 .7± 4 .2 ) cm / s,P<0 .0 0 1) ]。彩色多谱勒血流参数 PVV、HA- PI、肝血管指数对肝硬化门脉高压的诊断的敏感性和特异性分别为 89%、84 %、83%、81%、95%、91%。结论 :肝血管指数对肝硬化门脉高压的诊断的敏感性和特异性更高。     

Quantitative DNA measurement by flow cytometry and image analysis of human nonseminomatous germ cell testicular tumors

Current clinical staging, which includes the use of serum tumor markers and imaging techniques, fails to identify the 30–40% of clinical stage I (CS I) nonseminomatous germ cell testicular tumor (NSGCT) patients who have occult metastatic disease. Therefore, there is a real clinical need to evaluate new biological parameters of the primary tumor that might be useful as predictors of occult metastatic disease. This study was undertaken to compare quantitative DNA measurements by flow cytometry and image analysis in CS I NSGCT, and to analyze the relevance of these parameters for predicting occult lymph node involvement. Different blocks of formalin-fixed, paraffin-embedded NSGCTs of 62 CS I patients who underwent retroperitoneal lymph node dissection between 1985 and 1989 were prepared according to the Hedley technique, and analyzed by quantitative cytometry. Thirty-six (58.1%) patients had histologically proven lymph node involvement (pathological stage II), whereas 26 (41.9%) patients (pathological stage I) had neither lymph node metastases according to retroperitoneal lymph node dissection (RPLND) specimens nor tumor recurrence during follow-up. Concordant results were found in 76.5% of the samples by both cytometric techniques. For flow cytometry, the percentages of aneuploid cells in the S- and the G2M+S-phase were the most robust predictive parameters for lymph node involvement, whereas for image analysis the 5c exceeding rate (5cER) had the most predictive significance. Based on the experience obtained in this study, both cytometric techniques provide additional information on tumor aggressiveness that might be useful in therapeutic selection of early stage NSGCT patients for either RPLND or surveillance only.     

MACS检测胃癌腹腔冲洗液游离癌细胞的研究

目的 探讨胃癌腹腔微转移情况的检测方法及意义。方法 手术中切除肿瘤前收集腹腔冲洗液，采用磁激活细胞分离术（MACS）对不同病理分期胃癌患者腹腔冲洗液中癌细胞进行富集并检测。分别标记带磁珠的细胞角蛋白（CK）抗体，经磁柱富集CK^＋上皮细胞，用流式细胞仪检测其含量，并比较胃癌组与胃平滑肌瘤组（对照组）以及胃癌不同分期之间、磁富集前后CK^＋上皮细胞含量的差异。结果 在未经MACS富集的标本中较少发现CK^＋ CD45^-细胞；在富集后的标本中其含量在胃癌组与对照组有显著差异（41／50，1／10，P〈0．001）；pTNMⅠ～Ⅱ期与Ⅲ～Ⅳ期之间（0．67％，3．42％，P〈0．001）差异有非常显著性。结论 MACS能有效地富集上皮来源细胞，提高上皮源细胞的检出率，并能反映腹腔游离癌细胞数量；上皮细胞数量与胃癌的存在及临床病理分期有关，其有利于判断肿瘤转移和预后并指导治疗。     

Whole-Blood Assay to Measure Oxidative Burst and Degranulation of Neutrophils for Monitoring Trauma Patients

AbstractBackground and Purpose: Polymorphonuclear neutrophils (PMNs) protect the host from invading microorganisms, but excessive PMN activation after trauma causes tissue injury. Rapid monitoring of PMN function is critical for the assessment of the inflammatory state of trauma patients. Here, the authors adapted two simple and rapid methods to measure oxidative burst and degranulation of human PMNs in whole blood to avoid potential interference of cell isolation procedures with the assessment of PMN function.Material and Methods: Heparinized blood was drawn from healthy volunteers or trauma patients, preincubated at 37 °C for 5 min, and stimulated with N-formyl-methionyl-leucyl-phenylalanine (fMLP). Four assays for oxidative burst were tested: (1) cytochrome C; (2) homovanillic acid (HVA); (3) Amplex^® Red; and (4) flow cytometry with dihydrorhodamine 123 (DHR). PMN degranulation was assessed with flow cytometry using antibodies to: (1) CD11b/Mac-1 (CD18); (2) CD63; and (3) CD66b (CD67).Results: With the exception of the DHR method, all methods to measure oxidative burst were found to be unsuitable in whole blood due to interference of plasma proteins and hemoglobin with the fluorimetric or photometric readouts. By contrast, all degranulation methods were suitable for whole-blood studies. However, for the assessment of formyl peptide-induced degranulation, anti-antibodies to CD11b/Mac-1 and CD66b were up to five times more sensitive than antibodies to CD63. Thus, the degranulation and DHR methods were optimized for increased sensitivity, speed, and specificity and their usefulness to measure PMN function in trauma patients was tested.Conclusion: The whole-blood methods based on flow cytometry with DHR, anti-CD11b/Mac-1, and anti- CD66b are rapid, simple, and reliable techniques to assess PMN function for trauma research.     

Evaluation of blood flow patterns of true and false lumens in dissecting aneurysms using MR phase-contrast techniques

The magnetic resonance phase-contrast technique for the measurement of flow velocity and volume in true and false lumens was studied in six patients with chronic dissecting aneurysms. Phase-contrast images were obtained at a level perpendicular to the dissecting aneurysms of the descending aorta. As the maximum diameter of aneurysms increased, the ratio of the cross-sectional area of the false to the true lumen increased and the peak average velocity in the true lumen during systole was decreased. This technique proved invaluable for determining prognosis and operability for this condition.     

Contrast on T2-weighted images of the lumbar spine using fast spin-echo and gated conventional spin-echo sequences

A prospective study in 31 patients was designed to compare contrast quantitatively using axial conventional, gated spin-echo T2-weighted (T2W) (SE) (asymmetrical echo TE 30 and 80 ms) and axial dual-echo fast spin-echo (FSE) sequences (TE_eff20 and 120 ms) to image lumbar discs, nerve roots, and cerebrospinal fluid CSF. We used two quantitative measures, percent (%) contrast and contrast-to-noise ratio (CNR), to compare the sequences. The FSE sequence had greater % contrast and CNR on the first and second echo images for both disc and nerve root detection using these scan parameters. An axial FSE sequence, therefore, provided contrast characteristics similar to those of gated axial T2W SE sequence in the lumbar spine, with a 60% saving in acquisition time. The FSE sequence is now our standard axial T2W study for the lumbar spine.     

涎腺腺样囊性癌氩离子激光流式细胞术分析及其临床意义

为探讨流式细胞术对涎腺腺样囊性癌的诊断和预后估计的价值，对４１例腺样囊性癌行流式细胞术分析。结果，平均ＤＮＡ指数为１．１７９，Ｓ期细胞比例为２９．４９％，细胞增殖指数为４２．６７％，均明显高于正常组织。异倍体检出率为７０．７３％。随着肿瘤的增大和恶性程度的增加，其异倍体率、ＤＮＡ指数和Ｓ期细胞比例值也相应增加。流式细胞术为涎腺腺样囊性癌的病理诊断和确定恶性程度提供了有效的辅助诊断措施，但尚不能判断预后。