针刺抽吸法诱导建立椎间盘退行性变的动物模型

目的:用针刺抽吸法建立椎间盘退行性变动物模型,分析其可行性和优点。方法:实验于2006-10/2007-03在苏州大学附属第一医院骨科实验室完成。新西兰大白兔18只,用持针器夹持21G皮肤穿刺针从椎间隙正前方刺入L3～4、L4～5、L5～6椎间盘的纤维环,深度控制在5mm,拔出针芯,抽吸出部分髓核组织。术前及术后4,8,12周分别对造模后椎间盘及对照椎间盘(L2～3)行免疫组织化学及组织学观察。结果:新西兰大白兔18只均进入结果分析。①组织学观察:苏木精-伊红染色可见对照组髓核组织中有着大量髓核细胞,造模组第4周髓核细胞数量不断减少,第12周时髓核中主要为纤维软骨组织,伴有极少量髓核细胞。Ⅱ型胶原免疫组织化学检测可见:对照组髓核组织间质呈强阳性染色,而各模型组呈弱阳性染色。随着造模时间的延长,染色逐渐减弱,说明髓核组织中Ⅱ型胶原的含量逐渐减少。②髓核细胞凋亡率:对照组、实验组4,8,12周的髓核细胞凋亡率为(3±1)%,(17±4)%,(36±2)%,(23±5)%,差异有显著性意义(F=113.86,P=0.0001<0.01)。结论:纤维环穿刺吸出部分髓核的方法在不破坏自身免疫屏障的前提下可诱导兔椎间盘的缓慢退行性变,为干细胞、髓核细胞、基因治疗等提供有效的动物模型。     

医科大学就业大学生202名人格特征调查

目的:调查分析就业与未就业大学生的人格特质差异及就业的大学生人格特质是否有性别差异。方法:①纳入某医科大学临床、检验、放射、口腔、高护等专业毕业已就业的大学生和未就业的大学生。于2005-07用随机抽样的方式,选取其中录取的202名,其中男生91名,女生111名。未就业的大学生204名,其中男103名,女101名。纳入对象均健康,且对调查项目知情意。②采用加利福尼亚心理调查表进行个体水平、人格特征和自我实现倾向测试。该量表共440个测试题,20个分量表,3个结构量表。主要包括4大因素。人际关系和适应性,包括支配性、进取能力、社交性、社交风度、自我接受;社会化成熟感、责任心和价值感,包括责任心、社会化、自我控制、好印象、同众性、适意感、宽容性;成就潜能和智力水平,包括顺从成就、独立成就、智力效率;个人生活态度和倾向,包括心理感受性、灵活性、女性化/男性化。另外还有2个通俗概念量表即独立性、通情。3个结构量表内外向、常规趋向和常规异向和自我实现。分数越高表明心理素质越适应就业。③计量资料间差异性比较采用t检验。结果:共发出问卷406份,回收填写完整、准确的答卷406份,回收率100%。①就业与未就业大学生加尼福尼亚心理调查表分量表评分比较:就业大学生加尼福尼亚心理调查表分量表支配性、进取能力、社交性、社交风度、自我接受、独立性、通情、责任心、社会化、自我控制、好印象、同众性、适意感、宽容性、顺从成就、独立成就、智力效率、心理感受性评分明显高于未就业大学生(t=3.81～12.62,P<0.01)。②就业男女大学生加尼福尼亚心理调查表分量表评分比较:差异不明显(t=0.04～1.73,P>0.05)。③加尼福尼亚心理调查表结构量表评分比较:就业大学生常规趋向和常规异向和自我实现评分明显高于未就业大学生(t=5.00～10.06,P<0.01)。结论:毕业后能够就业的大学生人格特征较为成熟,较未就业大学更具备适应就业的心理素质和人格特质,此种人格特质无性别差异。     

重组结缔组织生长因子对体外培养成骨细胞核结合因子α1基因表达的影响

目的:研究发现,结缔组织生长因子可促进软骨细胞和成骨细胞的增殖及其表型的发生,在骨骼发育以及骨量维持方面发挥重要作用,但其对成骨细胞的作用机制目前尚不清楚。观察重组结缔组织生长因子对体外培养人成骨细胞核结合因子α1基因表达的影响。方法:实验于2006-01/2007-01在日照市人民医院完成。①实验材料:外科手术取正常成人髂骨松质骨,患者对试验知情同意。②实验方法:体外培养正常人成骨细胞,用不同浓度0,50,100,200,1000μg/L的重组结缔组织生长因子(0μg/L作为空白对照组)干预48h后,抽提细胞总RNA和总蛋白。③实验评估:采用半定量反转录-聚合酶链反应和蛋白免疫印迹分析方法观察不同浓度重组结缔组织生长因子对体外培养人成骨细胞核结合因子α1基因表达的影响。结果:半定量反转录-聚合酶链反应和蛋白免疫印迹结果显示,50,100,200,1000μg/L重组结缔组织生长因子干预后均可显著上调成骨细胞核结合因子α1的表达,并呈明显剂量依赖关系,与空白对照组比较,差异显著(P<0.01)。结论:重组结缔组织生长因子可剂量依赖性上调成骨细胞核结合因子α1基因的表达,核结合因子α1可能参与了结缔组织生长因子对成骨细胞增殖和分化的调节。     

贞芪扶正颗粒和复方阿胶浆干预再生障碍性贫血模型小鼠外周血及骨髓象的变化

目的:观察贞芪扶正颗粒和复方阿胶浆对苯油溶液致小鼠再生障碍性贫血模型的疗效。方法:实验于2005-06/09在河南中医学院药理实验室完成。①选用昆明种成年雄性小鼠90只。按随机数字表法将小鼠分为9组,每组10只:大、中、小剂量贞芪扶正颗粒组:按15,10,5g/kg剂量灌服贞芪扶正颗粒混悬液(主要成分:黄芪、女贞子;甘肃扶正药业科技股份有限公司生产;批号040803,15g/袋)。大、中、小剂量复方阿胶浆组:分别按10,20,30mL/kg剂量灌胃复方阿胶浆(主要成分:阿胶、熟地黄、党参、山楂、人参、蔗糖;山东东阿阿胶股份有限公司生产,批号050446,250mL/瓶),司坦唑醇组:按4mg/kg剂量灌胃司坦唑醇混悬液(司坦唑醇片,广西南宁百会药业集团有限公司生产,批号050306,30mg/片)。空白组和模型组:灌胃同体积的生理盐水(20mL/kg)。每天给药1次,连续给药14d。除空白组外,其余组小鼠皮下注射体积分数0.25苯的玉米油溶液4mL/kg复制苯油溶液致小鼠再生障碍性贫血模型,空白组皮下注射同体积玉米油。②各组小鼠分别于末次给药后24h,进行血常规检测。取右侧股骨,冲出骨髓细胞,采用BI-2000医学图像分析仪计数骨髓有核细胞数。③计量资料符合正态分布、方差齐者用t检验;方差不齐者用t’检验。结果:小鼠90只均进入结果分析。①血细胞测定结果比较:模型组小鼠血红细胞、白细胞、血小板计数和血红蛋白水平明显低于空白组(t=3.39～11.89,P<0.01)。司坦唑醇组3项血细胞计数和血红蛋白水平明显高于模型组(t=4.94～6.73,P<0.01)。大、中剂量贞芪扶正颗粒组和各剂量复方阿胶浆组血白细胞计数明显高于模型组(t=2.32～3.03,P<0.05￣0.01)。中剂量贞芪扶正颗粒组和各剂量复方阿胶浆组小鼠血红细胞计数明显高于模型组(t=2.15～4.84,P<0.05￣0.01)。大、中剂量贞芪扶正颗粒组和各剂量复方阿胶浆组血红蛋白水平明显高于模型组(t=2.33～4.45,P<0.05￣0.01)。大、中剂量贞芪扶正颗粒组和各剂量复方阿胶浆组血小板计数明显高于模型组(t=4.06～6.24,P<0.01)。②骨髓有核细胞数:模型组明显低于空白组(t=8.99,P<0.01)。司坦唑醇组、中剂量贞芪扶正颗粒组和小剂量复方阿胶浆组明显高于模型组(t=2.39～2.82,P<0.05)。结论:贞芪扶正颗粒、复方阿胶浆对皮下注射苯所致小鼠再生障碍性贫血模型血细胞状况及骨髓象均有较好的改善作用,以贞芪扶正颗粒有较为明显的剂量依赖关系。     

小鼠颅神经嵴细胞的培养和特征

目的：在体外原代培养Balb/c小鼠胚胎的颅神经嵴细胞。为颅面部各种组织细胞的发育研究提供细胞来源。方法：采用胰酶消化法分离小鼠胚胎第8.5天的颅神经管，从小鼠颅神经管中游离出来的细胞即为颅神经嵴细胞，用免疫组织化学方法鉴定细胞的来源，并测定细胞的生长曲线。结果：成功地培养出小鼠的颅神经嵴细胞，其形态类似成纤维样细胞，免疫组化检测结果表明，神经特异性烯醇化酶（NSE）抗体染色结果阳性。细胞的群体倍增时间为43.65h。结论：原代培养的小鼠颅神经嵴细胞生长稳定，来源明确，是颅面部各种细胞的发育和分化研究中一种有用的工具。     

Treatment of progressive Hodgkin's disease with intensive chemoradiotherapy and autologous bone marrow transplantation

Twenty-six patients with progressive Hodgkin's disease after conventional chemotherapy received intensive chemoradiotherapy and autologous bone marrow transplantation (ABMT); 19 also received additional involved-field radiotherapy. Twenty-one patients [81%, 95% confidence intervals (CI) 61% to 94%] attained complete (n = 18) or partial responses. Ten patients (38%, 95% CI 20% to 59%) are disease- free a median of 4.5 years later (range 3.5 to 7.0 years), including seven patients with continuous complete responses. The likelihood of overall response was not significantly influenced by any clinical or treatment variable examined. However, there was a trend favoring patients with higher Karnofsky scores, and higher scores were associated with attainment of complete responses (P = .06 and P = .02, respectively, Mann-Whitney U test). Both higher Karnofsky scores and shorter durations of disease before transplantation were associated with improved survival in a stepwise Cox multivariate analysis. The chief cause of failure was progression at sites previously involved with Hodgkin's disease. No patient relapsed in the marrow, and two of three patients with a history of marrow involvement with Hodgkin's disease achieved durable complete responses after transplantation. These data suggest that inadequate pretransplant conditioning, and not the reinoculation of occult tumor cells in the autologous marrow, caused most relapses. Fatal treatment-related toxicity occurred in six patients. Three patients died of idiopathic interstitial pneumonitis; each had previously received local mediastinal irradiation before intensive chemoradiotherapy. Intensive chemoradiotherapy and ABMT produces durable responses in some patients with Hodgkin's disease incurable with conventional therapy. Use of such therapies at the first sign of failure with conventional chemotherapy and development of more effective conditioning regimens should further improve results.     

Differential expression of CD11b/CD18 (Mo1) and myeloperoxidase genes during myeloid differentiation

During the course of differentiation of early human myeloid cells toward monocytes and granulocytes, cell surface expression of the cell adhesion molecule, CD11b/CD18 (Mo1) increases dramatically and expression of myeloperoxidase (MPO), a bacteriocidal enzyme, decreases markedly. Using the inducible promyelocytic cell line HL-60 as a model, we studied the mRNA expression of these genes. Differentiation of these cells along both a monocytic and a granulocytic pathway demonstrated that the mRNA levels of the two subunits of CD11b/CD18 increased in a pattern temporally and quantitatively similar to the increase in cell surface expression of this heterodimer. In contrast, the expression of MPO mRNA decreased in a temporal and quantitative pattern similar to the known decrease in MPO protein during differentiation, suggesting that regulation of these myeloid-specific proteins may occur at the level of mRNA expression. These findings have important implications with regard to the nature of the block in differentiation in acute nonlymphocytic leukemia and the regulation of myeloid gene expression.     

High-dose cytosine arabinoside and daunorubicin as consolidation therapy for acute nonlymphocytic leukemia in first remission: a pilot study

High-dose (HD) cytosine arabinoside (ARA-C) is more effective treatment than conventional-dose ARA-C regimens for patients with relapsed acute nonlymphocytic leukemia (ANLL). We report here that HD ARA-C given during the first remission of ANLL has resulted in long remission durations and a high proportion of patients who survive more than three years free of disease. From August 1979 to September 1983, 36 adult patients with ANLL in first remission received one to three courses of HD ARA-C (3 g/m2 by one-hour infusion every 12 hours for 12 doses on days 1 through 6) alone or with daunorubicin (30 mg/m2 for two or three doses on days 7 through 9). Three patients died of sepsis or hemorrhage during consolidation, and 14 patients have relapsed from five to 48 months after diagnosis. The remaining 19 patients are in continued complete remission (CCR) from 11 to 62 months. Denoting all deaths in remission as relapse, the actuarial probability of CCR is 42% at 62 months, with an apparent plateau in the survival curve. Of the first 22 patients treated, ten remain in CCR from 37 to 62 months with no therapy for at least three years. Due to its heightened anti-leukemic activity, HD ARA-C allows brief but effective consolidation of ANLL in first remission, with long-term disease-free survival comparable to other approaches.     

Relationship of the clinical response and binding of recombinant interferon alpha in patients with lymphoproliferative diseases

Patients with hairy cell leukemia (HCL) and chronic lymphocytic leukemia (CLL) were treated with recombinant interferon alpha A (rIFN- alpha A). The binding of iodinated recombinant interferon-alpha to baseline samples of peripheral blood mononuclear cells (PBMCs) from the leukemia patients was compared with clinical responsiveness to rIFN- alpha A. HCL patients (8/10) responded to rIFN-alpha A therapy, whereas none (0/10) of the CLL patients studied responded. The PBMCs from the eight responsive HCL patients bound approximately twice as much iodinated interferon as the PBMCs from nonresponsive CLL patients. This difference was due to more high-affinity receptors per cell with no difference in the affinity of the interferon-receptor interaction. However, because PBMCs from HCL patients were larger than PBMCs from CLL patients, the cell surface receptor density was similar. The leukemic cells from one of the two nonresponsive HCL patients bound iodinated interferon similarly to the cells from the responsive HCL patients, whereas the leukemic cells from the other nonresponsive HCL patient bound considerably less. The rapidity of response of the HCL patients did not correlate with the level of binding of iodinated interferon. Our results suggest that the absolute number of interferon receptors per cell may be only one of several important parameters in the response to rIFN-alpha A therapy, and that the responsiveness of a particular lymphoproliferative disease or a particular patient to rIFN- alpha A therapy cannot be predicted or explained solely by the degree of interaction between IFN and its cell surface receptor.