单核细胞趋化因子-1在间质性膀胱炎中表达的研究

目的 探讨单核细胞趋化因子-1(MCP-1)在间质性膀胱炎(IC)患者膀胱组织和尿液中的表达水平及其意义. 方法根据美国国立糖尿病、消化、肾病协会IC诊断标准确诊女性IC患者35例,感染性膀胱炎(UI)患者20例,肾囊肿患者25例作为正常对照组.IC患者平均年龄47(31～65)岁.主诉下腹酸胀/疼痛和夜尿次数多,伴有尿频尿急;已生育30例.均行24 h排尿卡记录、IC症状及问题评分表(O'Leary-Sant评分表)、钾离子敏感试验(PST)和麻醉状态下水扩张后膀胱镜检查.膀胱镜检查获取3组患者膀胱组织和尿液样本,免疫组化染色方法观察MCP-1在IC组织中的表达分布.RT-PCR技术测定3组膀胱组织中MCP-1 mRNA表达水平,酶联免疫吸附试验测定3组尿液标本中MCP-1水平. 结果 IC、UI、对照组尿液标本中MCP-1浓度分别为(74.1±36.9)、(280.6±68.9)、(10.8±6.9)pg/ml;膀胱组织中MCP-1相对定量值分别为76.2±24.0、99.5±30.1、36.1±14.1.lC组和UI组中组织/尿液中MCP-1表达水平均高于正常对照组,差异有统计学意义(P＜0.01).IC组临床症状评分为(14.9±1.8)分,与MCP-1升高水平呈正相关(r=0.686). 结论 IC患者膀胱组织和尿液中MCP-1表达升高,可能成为IC诊断的非特异性免疫指标之一.     

Monocyte chemoattractant protein-1 and interleukin-8 levels in children with acute poststreptococcal glomerulonephritis

The infiltration of leukocytes into the glomeruli is a major factor in inflammatory glomerular damage in acute poststreptococcal glomerulonephritis (APSGN). Chemokines participate in leukocyte infiltration. The aim of the present study was to investigate the role of monocyte chemoattractant protein-1 (CCL2/MCP-1) and interleukin-8 (CXL8/IL-8) in APSGN with special emphasis on their role in the clinical course of renal disease. Twenty-one children with APSGN were studied. Serum and urinary CCL2/MCP-1 and CXL8/IL-8 levels were measured by ELISA. The relationships between urinary chemokines and the degree of proteinuria were investigated. Serum and urinary CCL2/MCP-1 levels were significantly higher in the acute phase than in the resolution phase and in controls ( P <0.05). Urinary CCL2/MCP-1 levels in the control group were significantly lower than in both the acute and resolution phases ( P =0.01 and P =0.001, respectively). In the acute phase, urinary CCL2/MCP-1 correlated with the extent of proteinuria ( r =0.58, P =0.006) but not with serum CCL2/MCP-1 levels ( r =0.21, P =0.36). Urinary and serum CXL8/IL-8 levels were significantly elevated in the acute phase compared with the resolution phase and controls ( P <0.05). A consistent increase in urinary CCL2/MCP-1 was found in the acute phase of patients with APSGN, and this correlates with the degree of proteinuria. Our results emphasize the important role of locally produced chemokines in immune-mediated glomerular injury.     

单核细胞化学吸引蛋白质1 小分子干扰RNA人肾小管上皮细胞株的建立

目的 建立单核细胞化学吸引蛋白质1（MCP-1） 小分子干扰RNA（siRNA）人肾小管上皮细胞株，并检测MCP-1 siRNA对人肾小管上皮细胞（HKC）MCP-1基因的抑制效应。 方法 针对人MCP-1 mRNA 67、116、142位点设计并合成3对MCP-1 siRNA序列。构建MCP-1特异性siRNA真核表达载体，以脂质体法瞬时转染至HKC。转染24 h后分别应用实时定量RT-PCR及Western印迹技术检测HKC内MCP-1 mRNA、蛋白表达，筛选出抑制效率最高的MCP-1 siRNA。以筛选出的MCP-1 siRNA序列构建慢病毒穿梭质粒。使用慢病毒穿梭质粒进行慢病毒颗粒的包装和生产，得到病毒液并确定其滴度。以病毒液感染HKC，筛选MCP-1 siRNA人肾小管上皮细胞株，并应用实时定量RT-PCR及Western印迹技术检测HKC内MCP-1 mRNA、蛋白表达。 结果 成功建立MCP-1特异性siRNA人肾小管上皮细胞株。MCP-1 siRNA稳定转染能下调人肾小管上皮细胞MCP-1 mRNA (68.49±6.38)%、蛋白(72.97±6.13)%，与对照组比较差异有统计学意义(P < 0.01)。 结论 MCP-1特异性siRNA能高效抑制HKC内MCP-1基因表达。MCP-1 siRNA人肾小管上皮细胞株的建立为MCP-1基因功能及肾间质纤维化防治研究提供实验材料和依据。     

Monocyte chemoattractant protein-1 level in serum of patients with acute spinal cord injury

Spinalcordinjury(SCI)withinthefirstfewhours,isfrequentlycomplicatedbyinflammatorymechanisms,includingtheinfluxofmonocyte/macrophagesaswellastheactivationofresidentspinalmicrogliaandastrocytes.Numerousstudieshave suggestedthattheinitialinfiltrationofthe hematogenouscellsmaybeduetothesecretionof cytokinesandchemokinesintheinjuredcentral nervoussystem(CNS),amongwhichmonocyte chemoattractantprotein1(MCP1),amemberofβfamilychemokines,isthemostwellrecognized.Studiesconductedinmanylaboratorieshave…     

Olmesartan inhibits the expression of monocyte chemoattractant protein-1 and tumor necrosis factor-α and improves vascular remodeling after vascular injury in mouse

Vscularinjuryinducesmigrationofsmoothmusclecells (SMCs)fromthemediatotheintimaandsubsequent proliferationofSMCswithintheintima .Thesevascularresponsesareproposedtobecriticalprocessesofneointimalformationandinducenarrowingofthevascularlumenandarteriosclerosis .1,3Manystudieshavedemonstratedthatangiotensinconvertingenzyme(ACE)inhibitors4 ,5inhibitneointimalformationintheinjuredmodelofsmallanimal,whereasotheragentsthathaveblood pressuredeclinedfailtopreventit.4 Ininjuredarteries,theexpression…     

Role of immunologic costimulatory factors in the pathogenesis of biliary atresia

Background

The authors studied the patterns of expression of immunologic costimulatory molecules (B7-1, B7-2, and CD40) in biliary atresia (BA) patients to confirm any correlation with clinical course/outcome.

Methods

Based on clinical status 2 years postoperatively, 24 BA patients were divided into group I (n = 8, normal liver function), group II (n = 10, anicteric with moderate liver dysfunction), and group III (n = 6, icteric with severe liver dysfunction). Liver biopsies obtained at portoenterostomy and from 6 age-matched controls, were analyzed immunohistochemically using antibodies against B7-1, B7-2, and CD40.

Results

There was no expression of B7-1, B7-2, or CD40 in any control liver specimen. In all BA specimens, B7-1, B7-2, and CD40 were expressed strongly in bile ductules in portal tracts. In groups with liver dysfunction, B7-1, B7-2, and CD40 were expressed strongly on the surfaces of Kupffer and dendritic cells and in hepatocyte cytoplasm. Positive staining cells were significantly fewer in patients with better clinical outcome. B7-1 was found in vascular and sinusoidal endothelial cells only in cases of postoperative portal hypertension.

Conclusions

Costimulatory factors expressed on bile ductules, hepatocytes, and vascular endothelial cells appear to mediate autoimmune processes causing progressive liver fibrosis and portal hypertension in BA.     

Elevation of monocyte chemoattractant protein-1 in patients experiencing neurocognitive decline following carotid endarterectomy

Summary   Background. Previous studies have demonstrated that elevated pre-operative monocyte count is an independent predictor of acute neurocognitive decline following carotid endarterectomy (CEA). Monocyte chemoattractant protein-1 (MCP-1), secreted by human endothelial and monocyte-like cells, is a potent mediator of inflammation and mononuclear cell trafficking. This study examines the relationship between peri-operative serum MCP-1 elevation and post-operative neurocognitive injury following CEA. Methods. Fifty-two patients undergoing CEA and 67 lumbar laminectomy (LL) controls were administered a battery of five neuropsychological tests pre-operatively and on post-operative day 1 (POD 1). Change in individual test scores from baseline to POD 1 were converted into Z-score and used to develop a point system quantifying the degree of neurocognitive dysfunction relative to change within the LL group. Neurocognitive injury following CEA was defined as a score greater than 2 standard deviations above mean total deficit scores of LL controls. Serum MCP-1 levels were measured pre-operatively and on POD 1 by enzyme-linked immunosorbent assay. Findings. Mean percent MCP-1 elevation was higher for the 13 injured CEA patients (147.7 ± 32.4%) in our cohort compared to 39 age- and sex-matched uninjured CEA patients (76.0 ± 16.5%). In unconditional multivariate logistic regression analysis, percent elevation in serum MCP-1 level was associated with neurocognitive injury one day after CEA (OR = 2.19, 95% CI = 1.13–4.26, P = 0.021, for a 100% elevation from pre-operative levels). Conclusions. Peri-operative elevations in serum MCP-1 levels correlate with acute neurocognitive dysfunction following CEA. These data implicate an inflammatory mechanism in the pathogenesis of Ischaemic neurocognitive decline. Correspondence: Ricardo J. Komotar MD, Department of Neurosurgery, Columbia University, 710 West 168th Street, Room 431, New York, NY 10032, USA.     

单核趋化蛋白-2中和抗体对骨癌痛大鼠脊髓背角磷酸化细胞外信号调节蛋白激酶表达的影响

目的 观察鞘内注射单核趋化蛋白-2(monocyte chemoattractant protein-2,CCL2)中和抗体后骨癌痛大鼠行为学的变化,并探讨其可能的镇痛机制. 方法 雄性SD大鼠32只,体重180～220 g,采用随机数字表法分为4组(每组8只):假手术+正常IgG组(I组),于大鼠左侧胫骨干骺端骨髓腔内注射10μl Hank's液;假手术+CCL2中和抗体组(Ⅱ组),于大鼠左侧胫骨干骺端骨髓腔内注射10μl Hank's液,在注射Hank's液后第7～9天鞘内注射CCL2中和抗体,每天1次,每次10μl,浓度为103 mg/L;骨癌痛+正常IgG组(Ⅲ组),于左侧胫骨干骺端骨髓腔内注射10 μl(1×107/ml)Walker 256肿瘤细胞;骨癌痛+CCL2中和抗体组(Ⅳ组),在注射肿瘤细胞后第7～9天鞘内注射CCL2中和抗体,每天1次,每次10μl,浓度为103 mg/L.观察造模前及术后1、3、6、7、8、9d大鼠机械缩足反射阈值(mechanical withdraw threshold,PMWT)和热缩足反射潜伏期(thermal withdraw latency,TWL)及脊髓背角磷酸化细胞外信号调节蛋白激酶1/2(phosphorylate extracellular signal-regulated kinases 1/2,p-ERK1/2)的表达. 结果 与Ⅰ组比较,Ⅲ组大鼠的PMWT和TWL在第6～9天明显下降,脊髓背角p-ERK1/2蛋白表达明显增加(P＜0.01);与Ⅲ组比较,Ⅳ组大鼠的PMWT和TWL在第6～9天明显上升(P＜0.01),脊髓背角p-ERK1/2蛋白表达明显下降(P＜0.01);Ⅰ组和Ⅱ组上述指标比较,差异无统计学意义(P＞0.05). 结论 鞘内注射CCL2中和抗体可以部分缓解骨癌痛大鼠的机械性和热痛觉超敏,这种效应可能与抑制p-ERK1/2的表达有关.     

氟伐他汀对UUO大鼠肾间质单核细胞趋化蛋白-1表达及巨噬细胞浸润的影响

目的观察氟伐他汀对单侧输尿管梗阻大鼠肾间质单核细胞趋化蛋白-1(MCP-1)表达和巨噬细胞浸润的影响,探讨其抗纤维化机制。方法90只SD雌性大鼠随机分成假手术(SOR)组、单侧输尿管梗阻术(UUO)模型组和UUO+氟伐他汀治疗组(T-UUO,氟伐他汀20mg·kg-1·d-1)。于术后第1、4、7、10、14d分别处死各组大鼠。用HE及Masson染色动态观察肾脏病理变化,免疫组织化学法测定MCP-1、单核巨噬细胞抗原(ED-1)的表达。结果UUO模型组肾小管-间质MCP-1与ED-1表达较SOR组增加(P<0.05);在术后各时间点,T-UUO组大鼠肾小管-间质MCP-1、ED-1的表达及肾间质胶原相对面积较UUO模型组显著减少,但/仍高于SOR组(P<0.05)。结论氟伐他汀可通过降低MCP-1表达、减少单核/巨噬细胞浸润以抑制肾间质纤维化。     

胆道闭锁患儿肝移植术后门静脉狭窄的血管腔内治疗

目的探讨血管腔内治疗在胆道闭锁患儿肝移植术后门静脉狭窄(PVS)治疗中的应用价值。方法收集因原发病为胆道闭锁接受肝移植、术后后发生PVS的患儿14例,均经门静脉造影证实,并接受经皮血管成形术和(或)经皮血管内支架成形术治疗。分析14例患儿血管腔内介入治疗的效果。结果 14例患儿共进行23次血管内腔内介入治疗,技术成功率82.61%(19/23)。10例患儿经1~2次球囊扩张治疗后治愈,4例患儿球囊扩张治疗后,行血管腔内支架成形术,支架植入后未发生狭窄。14例患儿均未出现治疗相关并发症。结论胆道闭锁患儿肝移植术后PVS的血管腔内介入治疗安全、有效。     

胆道闭锁肝纤维化无创性诊断的研究进展

胆道闭锁是儿童进行肝移植的最常见的胆源性肝脏疾病,进行性肝纤维化是本病的显著特点,即使进行了肝门空肠R-Y吻合术,大多数患儿术后仍不可避免的出现肝纤维化的进行性加重,直至发展成为肝硬化、肝衰竭,并出现一系列并发症.因此肝纤维化评估是胆道闭锁患儿术后随访的重要内容,准确了解肝纤维化程度,对胆道闭锁患儿的病情评估有着重要的意义.肝组织活检是判断肝纤维化分级的金标准,但是其本身存在许多问题.运用多项无创性指标对患儿肝纤维化程度进行评估成为目前胆道闭锁预后研究的热点,本文对主要应用于胆道闭锁患儿肝纤维化的若干无创性诊断进行系统性分析,并分别从影像学和血清学两个方面进行综述.     

Histologic oddities at the porta hepatis in biliary atresia

Purpose

Highly unusual histologic findings at the porta hepatis in 3 infants who underwent Kasai portoenterostomy for biliary atresia are reported.

Methods

Portoenterostomy was performed using a standard operative technique. Serial transverse sections of the excised portal plate were examined by light microscopy along with sections from the distal extrahepatic biliary remnants, gallbladder, and liver biopsy.

Results

Of 61 consecutive infants who underwent Kasai portoenterostomy for biliary atresia, 3 were found to have highly unusual histologic features at the porta hepatis. All had type 3 biliary atresia. Two had hilar biliary ductules lined in part by squamous epithelium, and the third had a focus of mature hyaline cartilage surrounded by perichondrium adjacent to biliary ductules. In each case, these unusual histologic features were localized to the porta hepatis in the region of the transected portal plate.

Conclusions

The presence of hyaline cartilage at the portal plate is likely to be an expression of defective morphogenesis, thus supporting the concept of disordered embryogenesis in the etiology of biliary atresia. Squamous epithelium within biliary ductules might also reflect a similar mechanism but could alternatively be an unusual metaplastic response to inflammation at this site.     

Efficacy of antisense monocyte chemoattractant protein-1 (MCP-1) in a rat model of mesangial proliferative glomerulonephritis

Abstract

Objective: The effects of inhibition of monocyte chemoattractant protein-1 (MCP-1) on a rat model of mesangial proliferative glomerulonephritis (MsPGN) were evaluated. Methods: The anti-Thy-1 MsPGN model was developed by intravenously injecting anti-Thy-1 monoclonal antibodies into rats, followed by an injection of mesangial cells transfected with antisense MCP-1 into the renal artery. Exogenous cells were detected by in situ hybridization. Rats (40 total) were randomly divided into five groups: SO (sham operation), TG (Thy-1 glomerulonephritis model), MC (non-transfected normal rat mesangial cell), BC (pLXSN empty vector or blank control), and AM (antisense MCP-1 transfection) groups. Effects of exogenous MCP-1 on urinary protein excretion rate, biochemical parameters, and pathological changes were evaluated. Expression of MCP-1 and transforming growth factor-β₁ (TGF-β₁) were detected by immunohistochemistry. mRNA expression of MCP-1, TGF-β₁, and CC chemokine receptor 2 (CCR2) were detected by RT-PCR. Results: Exogenous MCP-1 cDNA was successfully transfected into mesangial cells. Exogenous mesangial cells were detected in glomeruli by in situ hybridization. Glomerular mesangial cell proliferation, 24-h urinary protein excretion rate, mRNA expression of MCP-1, TGF-β₁, and CCR2, and protein expression of MCP-1 all decreased in the AM group as compared to the control group (p?<?0.05), but there was no significant difference in the expression level of TGF-β₁ protein. Conclusions: (1) Mesangial cells can be used as a vector to transfect exogenous genes into kidneys; (2) antisense MCP-1 decreases mesangial cell proliferation and pathological injury in MsPGN model rats by decreasing expression of MCP-1 and CCR2; and (3) antisense MCP-1 suppressed mesangial cell proliferation and matrix accumulation in anti-Thy-1 MsPGN model rats, which did not entirely depend on TGF-β_1.     

The extent of biliary proliferation in liver biopsies from patients with biliary atresia at portoenterostomy is associated with the postoperative prognosis

Background/Purpose

In biliary atresia (BA), a derangement in the biliary system remains, despite portoenterostomy performance. Many factors can influence the disease progression rate. This study aimed to analyze the association between biliary proliferation extent in biopsies from BA patients and postoperative prognosis.

Methods

Biliary proliferation was evaluated by a morphometric analysis of the cytokeratin 7 positivity percentage (PCK7) in wedge liver biopsies from 47 BA patients. The extent of fibrosis was evaluated by a fibrosis score (FS). The outcome 1-year native liver survival was correlated, using a multivariable regression analysis, with PCK7, FS, and age at portoenterostomy.

Results

The PCK7 ranged between 0.80% and 14.79% (M ± SD = 7.36% ± 4.15%). Patients who died or underwent transplantation had higher PCK7 than survivors with their native livers (P < .001). The area under the receiver operating characteristic curve for PCK7 in relation to the outcome was 0.845 (P < .001). The cutoff point of PCK7 for the maximal effect on postoperative prognosis was 10.18% (sensitivity = 0.71, specificity = 0.88). The PCK7 was the only studied variable associated with 1-year native liver survival, independently of age and FS (P = .002).

Conclusion

The extent of biliary proliferation at portoenterostomy, evaluated by PCK7, was associated with 1-year native liver survival of BA patients.     

单核细胞趋化蛋白1 mRNA在颅内动脉瘤瘤壁内的表达

目的 探讨脑动脉瘤发生发展的病理过程。方法 对2例未破裂的颅内动脉瘤和11例破裂的颅内动脉瘤瘤壁组织进行常规HE染色和原位杂交的方法，观察单核细胞趋化蛋白1（MCP－1）的mRNA在瘤壁内表达的位置；1例正常脑动脉壁组织做对照。结果 2例未破裂和10例破裂动脉瘤壁组织HE染色，显示瘤壁仅由增厚的内膜和结缔组织外膜组成。纤维增厚的内膜有长梭形的成纤维细胞不规则排列。瘤壁全层有单核细胞浸润。1例破裂动脉瘤瘤壁仅存玻璃样变纤维结构，几乎不存在细胞成分。1例未破裂动脉瘤和8例破裂动脉瘤瘤壁有附属血栓，血栓呈机化表现。原位杂交结果显示：正常动脉未见杂交信号。2例未破裂和10例破裂动脉瘤瘤壁内有杂交阳性细胞，多为成纤维细胞，颗粒沉积于细胞质。单核细胞胞质少，阳性颗粒不明显。杂交阳性信号间断分布在动脉瘤瘤壁内膜，多出现于排列紊乱的成纤维细胞，淋巴细胞聚集处。1例破裂动脉瘤瘤壁仅存玻璃样纤维结构，几乎不存在细胞，未见杂交信号。动脉瘤附壁血栓内MCP－1 mRNA表达细胞有成纤维细胞、浆细胞、微血管的内皮细胞，表达部位在细胞质。结论 未破裂的和破裂的动脉瘤瘤壁的病理表现和MCP－1 mRNA的高表达，提示脑动脉瘤的发展是单核细胞为主的不断加强的慢性炎性过程。     

Corticosteroid therapy in biliary atresia

Sixteen patients with biliary atresia had 44 steroid courses for treatment of cholangitis or diminution of bile flow following Kasai hepatic portoenterostomy operations. A "blast" type (high dose/short duration) steroid administration was employed to potentiate the choleretic and anti-inflammatory effect. There was a significant augmentation of bile flow and a reduction in maximum temperature, serum bilirubin, and alkaline phosphatase.     

Connective tissue growth factor expression is increased in biliary epithelial cells in biliary atresia

Background/Purpose

Connective tissue growth factor (CTGF) has been implicated in the pathogenesis of hepatic fibrosis and is elevated in the serum of children with biliary atresia (BA). The objective of this study was to evaluate hepatic CTGF messenger RNA (mRNA) expression and its relationship to hepatic histology in children with BA.

Methods

Connective tissue growth factor mRNA expression was evaluated by in situ hybridization in 26 liver biopsies from 11 patients with BA, 11 with other diseases, and 4 autopsy controls. Serial sections were immunostained with cell-specific markers to characterize the cells expressing CTGF. Biopsies were scored for CTGF expression (0-4) and inflammation and fibrosis (1-4).

Results

High levels of CTGF expression were observed in 9 of 11 BA with localization to biliary epithelial cells and vascular endothelial cells. Connective tissue growth factor mRNA expression was correlated with fibrosis in BA and all livers. In the 11 patients with other liver diseases, 7 had CTGF expression limited to hepatic stellate cells and vascular endothelial cells. None of the 4 livers in children without liver disease had significant levels of CTGF.

Conclusions

In BA livers, novel biliary epithelia CTGF mRNA expression is high and correlates with severity of fibrosis. These data support a role for biliary epithelial cell signaling in fibrogenesis.