Monocyte adhesion molecule expression in interstitial inflammation in patients with renal failure.

BACKGROUND: Patients with renal failure have an increased susceptibility to infections. We therefore studied the recruitment of monocytes and their expression of adhesion molecules CD11b and CD62L at the site of interstitial inflammation in patients with renal failure. Furthermore, we studied if the capacity of monocytes to up-regulate CD11b in interstitial inflammation was determined by the interstitial concentration of chemotactic factors. METHODS: Three intensities of interstitial inflammation (0, intermediate and intense) were established in skin blister chambers. Leukocyte count, CD11b/CD62L expression, monocyte chemotactic protein-1 (MCP-1) and blister activity in terms of CD11b mobilization were determined. RESULTS: The CD62L expression on monocytes was lower in the peripheral circulation in patients with renal failure compared with healthy subjects (P<0.005 and P<0.001). At the site of interstitial inflammation patients had a higher expression of CD62L (intermediate, P<0.05; intense, P<0.005). Furthermore, monocytes from patients had an impaired capacity to mobilize CD11b both in the peripheral circulation (P<0.005) and at the intermediate and intense sites of interstitial inflammation (P<0.005 and P<0.001, respectively) compared with cells collected from healthy subjects. We incubated monocytes in blister exudates, in order to explore whether this phenomenon is caused by cellular factors and/or to the interstitial concentration of chemotactic mediators. The expression of CD11b on monocytes from healthy blood donors incubated in blister exudates from either patients or healthy subjects in vitro was similar. The interstitial concentration of MCP-1 at the site of intermediate inflammation was significantly lower in patients with renal failure compared with the corresponding blister exudate collected from healthy subjects (P<0.05), but no differences were observed at the site of intense inflammation. Furthermore, neutralizing the action of MCP-1 in blister exudates with monoclonal antibodies did not have any impact on monocyte CD11b expression following incubation in blister exudates. CONCLUSION: These studies indicate that the impaired capacity of monocytes to mobilize CD11b at the site of inflammation in patients with renal failure is more dependent on constitutive cellular factors than the concentration of CD11b mobilizing factors in the interstitium.     

Intravenous drug abuse and Type 1 diabetes: financial and healthcare implications.

AIMS: To determine the morbidity, mortality and healthcare costs of intravenous drug-abusing patients with Type 1 diabetes (IVDA-DM), who are admitted to hospital. METHODS: Retrospective case note analysis of admissions, complications and cost estimation over a 6-year period. Each drug-abusing patient (IVDA-DM) (n = 9) was compared with two controls (n = 18) with Type 1 diabetes but without a history of intravenous drug abuse (DM-controls). Admissions were also analysed for patients with intravenous drug abuse, but without Type 1 diabetes (IVDA-controls) (n = 198). Admissions were at a University teaching hospital in Liverpool, UK. DM-controls were drawn from a population attending diabetes outpatient clinics between 1997 and 2002 at the same hospital. The main outcome measures were: the duration and healthcare costs of hospital admissions per year, outpatient attendances per year, glycated haemoglobin (HbA(1c)), weight, micro- and macrovascular complications and mortality. RESULTS: Multiple admissions, mainly related to ketoacidosis, led to marked differences in mean (95% CI) inpatient days per year per patient [IVDA-DM 28.1 (13.6-42.7) vs. DM-control 1.1 (0.2-1.9); P < 0.0001], mean inpatient days per year per patient in critical care bed (IVDA-DM 1.7 (-0.7-4.2) vs. DM-control 0; P < 0.02) and mean costs of admission, per patient per year (pound sterling 7320 vs. pound sterling 230). The IVDA-DM group frequently omitted insulin, were underweight, failed to attend as outpatients and five had died by the end of 2002. The IVDA-controls spent considerably less time in hospital [3.4 (2.8-3.9) days per patient per year]. CONCLUSION: IVDA-DM patients have higher rates of diabetes complications, are admitted more frequently and have a high mortality compared with DM and IVDA-controls. The cost of inpatient care of this small group of patients was considerable.     

多效蛋白在大肠癌恶性化过程中的作用

目的了解多效蛋白在大肠癌中的表达情况 ,以及对大肠癌恶性化的影响。方法采用RT PCR和免疫组化染色 ,对 2 4例大肠癌组织和 19例大肠癌癌旁组织的多效蛋白mRNA表达情况进行分析。结果 2 4例大肠癌组织和 19例癌旁正常组织中表达多效蛋白mRNA的各有 18例。多效蛋白不但表达在肿瘤细胞 ,还表达在其他间质细胞中。大肠癌组织中多效蛋白的表达明显高于癌旁正常组织 (34%vs.9% ,P <0 0 5 )。 (94 % )大肠癌多效蛋白mRNA主要由内源性多效蛋白转录 ,且表达在肿瘤的 4个分期中 ,而人类内源性逆转录病毒 多效蛋白仅融合转录在Ⅲ、Ⅳ期肿瘤上 (31% )。结论多效蛋白尤其是人类内源性逆转录病毒 多效蛋白与大肠癌的恶性化有关。     

生长激素抗肝纤维化的作用及其机制

目的　探讨重组人生长激素 (rhGH)对肝硬化大鼠在肝功能、门静脉高压等方面的治疗作用及其可能的机制。方法　雄性SD大鼠建立肝硬化模型 ,随机分组 ,分别给予NS、rhGH (333ng/KgBW ,ip ,qd× 7d)处理。 结果　rhGH处理前后肝硬化肝组织GHR的配体结合容量 [fmol/mg ,(31± 4 )vs .(4 0± 7) ]及其mRNA量 (iOD ,pixel)均显著升高 (2 3± 3)vs.(4 2± 8) ;P <0 0 5 ,肝硬化大鼠血清白蛋白 (g/L )显著升高 (2 9± 4 )vs .(37± 7) ;P <0 0 5、ALT活性 (U/L)显著降低 (89± 15 ,6 9± 7;P <0 0 5 ) ,肝硬化肝组织丙二醛含量 (nmol/mg)显著降低 (18 7± 3 2 ,12 0± 2 2 ;P <0 0 5 )、超氧化物歧化酶活性 (U/mg)显著升高 (82 4± 10 8,10 2 9± 76 ;P <0 0 5 )、胶原纤维相对含量 (% )显著降低 (2 2 30± 3 86vs.14 70± 2 0 7;P <0 0 5 ) ,肝硬化大鼠的门静脉压 (cmH2 O)亦显著降低 (14 4± 2 0vs.9 3± 1 5 ;P <0 0 5 )。结论　药理剂量的rhGH能够促进肝硬化大鼠的白蛋白合成、改善肝细胞功能 ,有助于抑制肝组织纤维化、缓解门静脉压力。     

骨髓细胞移植上调血管内皮生长因子及其受体的表达并改善缺血心脏功能

目的：通过骨髓细胞移植对心肌梗死大鼠模型血管内皮生长因子及其受体表达的影响，探讨骨髓细胞移植改善缺血心脏功能的可能机制。方法：利用左前降支冠状动脉结扎术制备大鼠心肌梗死模型，然后行异体骨髓细胞移植；分别于术后1d，3d，7d，14d和28d取材；利用免疫荧光和RT—PCR技术分析细胞移植对血管内皮生长因子(VEGF)及其受体(Flk一1)表达的影响和移植细胞的分化情况；通过免疫组化计算血管数量；应用血流动力学检测大鼠心脏功能在术后各时间点的变化。结果：VEGF和Flk-1在移植组动物的心肌梗死区残存细胞、梗死周边区细胞以及部分移植细胞内表达。移植组VEGF和Flk-1的mRNA表达明显高于对照组。分别于术后3d和14d达到高峰，以后逐渐减弱。术后7d，14d和28d移植组血管数量较同期对照组明显增加，移植组心脏功能较对照组明显改善。术后14d可在部分移植细胞中检测到心肌细胞或血管内皮细胞特异性蛋白的表达。结论：骨髓细胞移植通过上调移植细胞及受者内源性VEGF和Flk—1的表达，促进血管新生，进而改善缺血心脏功能。     

Constitutive secretion of the granule chymase mouse mast cell protease-1 and the chemokine, CCL2, by mucosal mast cell homologues

BACKGROUND: The mucosal mast cell (MMC) granule-specific beta-chymase, mouse mast cell protease-1 (mMCP-1), is released systemically into the bloodstream early in nematode infection before parasite-specific IgE responses develop and TGF-beta1 induces constitutive release of mMCP-1 by homologues of MMC in vitro. Intraepithelial MMC may also express the chemokine CCL2 (monocyte chemotactic protein-1) during nematode infection but the expression of this chemokine by MMC homologues has not been investigated. OBJECTIVE: To investigate the expression and to compare the mechanisms of constitutive release of the chymase, mMCP-1, and the chemokine, CCL2. METHODS: MMC homologues were generated by culturing bone marrow cells in the presence of TGF-beta1, IL-3, IL-9 and stem cell factor (SCF). The intracellular distribution of mMCP-1 and CCL2 was examined by confocal microscopy. The involvement of the Golgi complex and of protein synthesis in the constitutive release of mMCP-1 and CCL2 was investigated using the Golgi-disrupting agent brefeldin A and cycloheximide to block protein synthesis. Secreted analytes were quantified by ELISA. RESULTS: mMCP-1 colocalized with Golgi matrix protein 130 but was most abundant in the granules, whereas CCL2 was not found in the granules but appeared to be located uniquely in the Golgi complex. Extracellular release of mMCP-1 was significantly inhibited ( approximately 40%) by cycloheximide and by the Golgi-disrupting agent brefeldin A, indicating both continuous protein synthesis and transportation via the Golgi complex are required for optimal mMCP-1 secretion. A similar but more marked inhibitory effect with both compounds was demonstrated on the constitutive secretion of CCL2. CONCLUSION: The culture conditions that promote mMCP-1 expression and release by MMC homologues also promote the expression and release of CCL2. Constitutive release involves de novo protein synthesis and requires a functional Golgi complex, suggesting that similar mechanisms of extracellular secretion operate for both mediators.     

METH1在酵母双杂交系统中的表达及对报告基因激活作用的检测

目的通过观察血管生成抑制因子METH1的cDNA片段在酵母双杂交中的表达及检测其对报告基因有无激活作用，为进一步明确METHI抑制增生性瘢痕的分子机制奠定基础。方法采用酵母双杂交Ga14系统3，经PCR扩增子METH1的cDNA片段，分别克隆入pUC19质粒，经测序正确后。再分别亚克隆入酵母双杂交诱饵载体pGBKT7中。将重组质粒导入酵母菌AH109，检测其表达产物在酵母细胞中对报告基因的激活作用。结果成功获得METH1的cDNA片段，该片段所表达的蛋白对酵母菌AH109无毒性，且对报告基因无激活作用。结论血管生成抑制因子METH1蛋白活性区在酵母双杂交系统中的表达产物。可作为诱饵蛋白进行相互作用蛋白的筛选研究。     

前列腺癌组织中性激素受体表达的分析

目的:定性分析雄激素受体(AR)、雌激素受体(ER)、孕激素受体(PR)在良性前列腺增生(BPH)和前列腺癌(PCa)组织中的阳性表达,为激素治疗PCa和预后判断提供一定的依据。方法:采用免疫组化DAKO Envision二步法,分析30例BPH与32例PCa标本中AR、ER、PR的阳性表达情况。结果:AR、ER、PR阳性数BPH组分别为22、10、6例;PCa组分别为18、14、2例,两组差异无显著性。随访30例PCa患者术后的生存期表明:AR表达阳性者平均生存时间为6.41年,AR阴性者为4.28年,两者差异有统计学意义(P<0.05)。结论:BPH和PCa患者在性激素受体表达上的差异无显著意义;PCa患者中AR表达阳性者平均生存时间长于AR阴性者,后者的预后较差,对AR阴性者行激素治疗效果不确定。     

碱性成纤维细胞生长因子转染兔关节软骨细胞的研究

目的　探讨碱性成纤维细胞生长因子 (bFGF)基因转染兔关节软骨细胞后对培养的关节软骨细胞形态、分裂增殖及代谢等方面的影响。方法　将bFGF基因克隆于真核表达载体pHβ。AP 1中 ,构建重组真核表达载体 pHβ bFGF ,转染兔关节软骨细胞。G418筛选阳性克隆 ,检测阳性细胞bFGF基因的表达水平。测定培养软骨细胞的DNA含量、糖醛酸含量、软骨细胞增殖情况及进行细胞周期分析。结果　bFGF基因转染软骨细胞表型未见显著变化 ;bFGF基因转染组、载体对照组、空白对照组DNA含量分别为 ( 77.37± 6 .2 1)、( 40 .39± 4.33)、( 33 .77± 4.2 5 ) μg/瓶 (P <0 .0 1) ,糖醛酸含量分别为 ( 30 8.8± 10 .2 )、( 77.9± 8.7)、( 80 .2± 10 .5 ) μg/瓶 ( P <0 .0 1) ,软骨细胞G1期分别为 5 9.3± 2 .1、6 9.5± 4.0、73 .1± 3 .9(P <0 .0 5 )。结论　bFGF转染关节软骨细胞后 ,可显著促进细胞分裂增殖并缩短细胞周期 ,为软骨组织工程研究提供新的技术路线及理论基础。     

Expression of the calcium receptor CaR in the parathyroid of secondary hyperparathyroidism patients

The effects of calcium on parathyroid hormone (PTH) has further discovered in recent years. It has been known that calcium ion concentration in the extracellular fluid is a major determinant of PTH secretion. The relationship between serum intact PTH (iPTH) and calcium ion levels is described by a sigmoidal curve. The calcium concentration that produces half-maximal change in PTH release (the midpoint between maximal and minimal