大鼠坐骨神经损伤修复后近端轴突再生的初步研究

目的 采用生长相关蛋白-43(GAP-43)对SD大鼠坐骨神经近端新生轴突进行标记,观察坐骨神经损伤修复后近端轴突再生过程.方法 64只SPF级SD大鼠于右侧坐骨神经分叉以上5mm处切断坐骨神经,分别采用原位缝合以及生物可溶性甲壳质套管小间隙(2 mm)缝合方法进行修复,分别于术后1、3、7及14 d取材.观察神经缝合处组织大体形态;套管内坐骨神经生长状态;近端新生轴突形态,并应用图像分析方法对新生轴突数目进行定量分析.结果 套管小间隙缝合组大鼠神经缝合处粘连情况较轻.免疫荧光染色显示:术后14 d,新生纤维神经干形成圆锥形向前生长.形态规则均一.原位缝合组大鼠缝合处粘连较重,新生轴突于7 d左右长过缝合点,坐骨神经远端新生轴突散在分布,形态不规则.免疫荧光染色图像分析结果显示:大鼠坐骨神经损伤修复后,术后3 d原位缝合组新生轴突数目较套管缝合组多.差异有统计学意义(P<0.01);术后7 d套管小间隙缝合组大鼠坐骨神经开始大量再生;术后14 d左右,套管小间隙缝合组新生轴突数目显著高于原位缝合组(P<0.05).结论 甲壳质套管具有良好的生物相容性,套管内新生轴突前沿以圆锥形向前生长,新生轴突形态较原位缝合组好,套管小间隙修复7 d后新生轴突数目开始较原位缝合组多.本实验主要观察了两种术式修复后大鼠坐骨神经新生轴突的生长规律,也从组织学角度解释了生物套管小间隙套接方法的再生效果比原位缝合效果好的可能原因所在.     

复方聚乙二醇电解质散与番泻叶在结肠镜检查前肠道准备中的应用

为观察复方聚乙二醇电解质散与番泻叶联合应用在结肠镜检查前肠道准备的临床效果,将复方聚乙二醇电解质散与番泻叶联合应用为观察组和单用复方聚乙二醇电解质散为对照组,观察术前的排便次数和粪便性状、术中的肠道清洁满意度。结果显示．观察组排便结束时间明显短于对照组（P〈0．05）。肠道清洁满意度观察组为80．6％（162／201）,对照组为69．6％（133／191）（P〈0．05）。结果表明,复方聚乙二醇电解质散与番泻叶联合应用在结肠镜检查前肠道准备的效果比单用复方聚乙二醇电解质散效果好。     

肝细胞性肝癌生长激素受体的基因分析

目的 对肝细胞性肝癌生长激素受体(GHR)的表达进行基因分析.方法 分别采用放射配体法、半定量的逆转录-多聚酶链式反应(RT-PCR)对40例肝细胞性肝癌(HCC)组织进行GHR的检测,以8例正常肝组织作为对照,并随机选择15例RT-PCR阳性扩增产物直接进行DNA序列测定.结果 在蛋白表达水平,应用放射性配体法于正常肝组织与34例HCC组织中检测到GHR,对照组肝组织GHR的RT值为(40.2932±3.5667)fmol/mg·蛋白,K_d为(0.6167±0.1007)nmol/L;HCC组织GHR的RT为(19.8870±3.7549)fmol/mg·蛋白,K_d值为(0.6247±0.2011)nmol/L,与正常肝组织比较,HCC组织GHR的RT降低(P<0.05),而K_d无显著性改变(P>0.05),有6例肝癌组织在蛋白水平未检测到GHR;半定量的RT-PCR法发现GHRmRNA在正常肝组织的表达率为100%,在HCC组织的表达率为90%(36/40),GHR mRNA在正常肝组织与HCC组织的表达量两者之间差异无统计学意义(P>0.05).结论 大部分肝细胞性肝癌组织在蛋白与mRNA水平均表达GHR,肝癌与正常肝组织GHR在蛋白表达水平上差异的原因可能并非由于两者在mRNA水平表达量的不同所造成.在肝癌受体功能未明的情况下,目前rhGH在肝癌患者中的使用应慎重,以免促进肿瘤的增殖与复发.     

Growth Differentiation Factor-5 Stimulates the Growth and Anabolic Metabolism of Articular Chondrocytes

Objective： To observe the effect of growth differentiation factor-5 （GDF-5） on the growth and anabolic metabolism of articular chondrocytcs. Methods： The articular chondrocytes isolated from rats were treated with various concentrations of rmGDF-5, and the growth of chondrocytes measured by MTT assay, the cellular cartilage matrices formation detected sulfated glycosaminoglycan by Alcian blue staining and type Ⅱ collagen by RT-PCR, the collagen phenotypic expression of chondrocytes detected by immunofluorescence. Results： After 7 days culture, MTT assay showed that GDF-5 enhanced the growth of chondrocytes in a dose-dependent manner, RT-PCR showed that GDF-5 clearly induced the synthesis of type Ⅱ collagen because of the col2al mRNA band more and more strong in a dose-dependent. Chondrocytes were cultured with GDF-5 for 14 days, the intensity of Alcian blue staining was greatly enhanced, especially, at a high concentration of 1000ng/ml, and GDF-5 enhanced the accumulation of the Alcian blue-stainable material in a concentration-dependent manner and in a does-dependent manner. Chondrocytes were cultured with GDF-5 for 21 days, immunofluorescent staining of type Ⅱ collagen was clear, the type Ⅰ and X collagen were negative. Conclusion： GDF-5 enhanced the growth of mature articular chon- drocytes, and stimulated the cellular cartilage matrices formation, but did not change the collagen phenotypic ex- pression of chondrocytes in mono-layer culture.     

Prostaglandin E1 and misoprostol increase epidermal growth factor production in 3D-cultured human annulus cells

Background contextEpidermal growth factor (EGF) is a peptide known to modulate a number of cellular responses including embryogenesis, cell proliferation, and cell survival. Little is known about EGF and its regulation in human annulus cells. Previous work has identified EGF and its receptor in control outer annulus disc tissue, but not in herniated tissue.PurposeTo determine if human annulus cells express EGF in vitro, to determine if the epidermal growth factor-receptor (EGF-r) was expressed in vivo and in vitro in disc cells, to test the effect of EGF on annulus cell proliferation and proteoglycan production in vitro, and to test the effect of prostaglandin E1 (PGE1) and misoprostol on disc cell production of EGF in vitro.Study design/settingStudies were approved by the authors' Human Subjects Institutional review Board. Human disc tissue was used for immunocytochemistry, and human annulus cells were tested in vitro.Patient sampleThirty-four disc specimens were used for studies of proteoglycan production, cell proliferation, and EGF production in vitro. An additional nine discs were used for EGF-r immunolocalization.MethodsDisc tissue was used for immunocytochemical studies for the localization of EGF-r and as a source for cultured annulus cells. Monolayer culture was used to test proliferation responses to 0, 25, 50, or 75 ng/mL EGF over a 2-day culture period. Three-dimensional (3D) culture in a collagen sponge was used to test 100,000 cells cultured in a paired experimental design over 14 days for production of EGF and proteoglycans. Cells were exposed to control conditions, or to either misoprostol at 8 ng/mL or PGE1 at 10^?7 M. Conditioned media was harvested and assayed using an enzyme-linked immunosorbent assay (ELISA) assay with the Human Protein Cytokine Antibody Array I kit. Replicate EGF relative intensity values were averaged and normalized to controls assayed on the same membrane. 3D-cultured cells were also used to confirm EGF gene expression using microarray analysis. Standard statistical methods were used to analyze results.ResultsMicroarray analysis of mRNA from annulus cells in 3D culture confirmed expression of EGF, and immunocytochemistry verified the presence of EGF-r in vitro and in vivo. PGE1, at a dose of 10^?7 M, and misoprostol (a synthetic PGE1 analog) at a dose of 8 ng/mL, both significantly increased EGF levels in annulus cells cultured in 3D compared with control levels (p=.031 and .034, respectively). No significant difference, however, was seen in cell proliferation or in total sulfated proteoglycan production in EGF-exposed annulus cells.ConclusionsData showed that EGF is expressed and produced by annulus cells in vivo and in 3D culture, with significantly greater in vitro EGF produced in the presence of PGE1 or the PGE1 analog misoprostol. Misoprostol, developed for prevention/treatment of nonsteroidal anti-inflammatory-induced gastropathy, has now been reported to have some interesting anabolic effects stimulating osteoblasts during fracture healing and during ovariectomy in animal models. Exogenous EGF did not increase cell proliferation in monolayer, or total production of proteoglycans in 3D culture. Additional work is needed to further delineate the role of EGF in the human disc.     

N-acetylglucosaminidase,myeloperoxidase and vascular endothelial growth factor serum levels in breast cancer patients

Inflammatory cells surround breast carcinomas and may act promoting tumor development or stimulating anti-tumor immunity. N-acetylglucosaminidase (NAG) has been employed to detect macrophage accumulation/activation. Myeloperoxidase (MPO) is considered a marker for neutrophils activity/accumulation. Vascular Endothelial Growth Factor (VEGF) is as strong pro-angiogenic cytokine. The aim of this study was to measure the systemic inflammatory response by measuring serum levels of NAG, MPO and VEGF in women diagnosed with breast cancer and associate this response to the peritumoral inflammatory infiltrate and to prognostic factors. Serum samples obtained from women with no evidence of disease (n = 31) and with breast cancer (n = 68) were analyzed for the activities of NAG, MPO and VEGF by enzymatic assay. Serum levels of NAG and VEGF were higher in healthy volunteers (P < 0.0001) and serum levels of MPO were higher in patients with breast cancer (P = 0.002). Serum levels of NAG were positively correlated to serum levels of MPO and VEGF (P < 0.0001 and P = 0.0012, respectively) and MPO and VEGF serum levels had also a positive correlation (P = 0.0018). The inflammatory infiltrate was not associated to serum levels of the inflammatory markers, and higher levels of MPO were associated to lymphovascular invasion negativity (P = 0.0175).     

复方甘草酸单铵注射液中甘草酸单铵鉴别方法的改进

目的：建立复方甘草酸单铵注射液中甘草酸单铵鉴别方法的改进方法。方法：取本品1ml（约相当于甘草酸单铵2mg）,加入20%三氯乙酸乙醇溶液1ml,置水浴蒸至近干,残留物显红紫色。结论：本方法的检测灵敏度较原方法高,可用于复方甘草酸单铵注射液中甘草酸单铵的鉴别。     

RP-HPLC同时测定复方碘苯甲酸涂剂（Ⅰ）中两种酸的含量

目的建立一种测定复方碘苯甲酸涂剂中苯甲酸和水杨酸含量的高效液相色谱方法。方法采用反相C18色谱柱,以甲醇-0.05mol·L^-1磷酸二氢钾溶液（50∶50）（磷酸调节pH至4.0）为流动相,流速1.0mL·min^-1,检测波长240nm。结果水杨酸线性范围0.8～45.2μg·mL^-1（r=0.9997）,高、中、低浓度的平均回收率分别为99.3%,99.7%,99.9%;苯甲酸线性范围1.5～90.2μg·mL^-1（r=0.9993）,高、中、低浓度的平均回收率分别为100.2%,99.7%,99.6%。结论该法分离度好,快速,简便,重现性好,可用于复方碘苯甲酸涂剂（Ⅰ）的质量控制。     

复方维生素U片崩解时限不合格的原因及对策

目的查找造成不合格药品的主要原因,筛选复方维生素U片最佳处方。方法考察羧甲基淀粉钠、低取代羟丙基纤维素和吐温-80对复方维生素U片的质量影响,并与2种市售品作崩解时限比较。结果经分析不合格原因后筛选的最佳处方制备的复方维生素U片．崩解时限显著缩短。结论优选处方合理、工艺简单,可供生产厂家参考。     

复方丹参滴丸治疗冠心病心绞痛60例疗效观察

目的探讨复方丹参滴丸治疗冠心病心绞痛的疗效,评价其用药的合理性和安全性。方法选取符合条件的冠心病心绞痛患者100例,随机分为治疗组和对照组,分别为60例和40例。治疗组用复方丹参滴丸,对照组用甘油三酯片,疗程均为60天,于治疗前后分别做心绞痛症状改善、心电图疗效和血液流变学等的连续观察。结果治疗组和对照组心绞痛症状改善的总有效率分别为88．3％和60．0％。心电图疗效改善的总有效率分别为71．7％和52．5％。治疗组明显高于对照组,差异有显著性。血液流变学指标的比较显示,治疗组在改善全血黏度的高切和低切,毛细管血浆黏度,红细胞压积,纤维蛋白原等方面均较对照组明显。结论复方丹参滴丸在治疗冠心病心绞痛方面效果明显。