氯氮平、氯丙嗪、利培酮对肝脏纤维化指标影响的研究

目的：了解抗精神病药物氯氮平、氯丙嗪、利培酮及其联合用药后对肝脏纤维化指标Ⅲ型前胶原（PⅢNP）、Ⅳ型胶原（Col-Ⅳ）、层黏连蛋白（LN）和透明质酸（HA）的影响。方法：选择211例服用氯氮平（93例）、氯丙嗪（47例）、利培酮（31例）或联合用药（40例）治疗超过三年的精神病患者作为对象,并选择性别、年龄相匹配的75例健康人群作为对照组,抽取外周血,用酶联免疫法检测其PⅢNP、Col-Ⅳ、LN及HA。结果：①与正常对照组比较,所有服药患者血清肝纤维化指标PⅢNP及HA存在显著差异（P〈0.01）,而Col-Ⅳ和LN则无差异;②氯氮平组、氯丙嗪组、利培酮组及联合用药组之间比较,血清肝纤维化指标均无显著差异;与对照组比较,单一用药组（氯氮平组、氯丙嗪组和利培酮组）PⅢNP及HA存在显著差异（P〈0.01）,而联合用药组则PⅢNP、HA及LN均存在差异。结论：氯氮平、氯丙嗪及利培酮均对血清肝脏纤维化指标PⅢNP和HA有影响;各药物对肝纤维化指标的影响无差异,但其联合用药对肝纤维化指标的影响更大。     

Differential localization of laminin gamma and integrin beta in primary cultures of the rat gingival epithelium

OBJECTIVES: The aim of this study was to investigate the differential immunolocalization of laminin gamma(2) and integrin beta(4) in primary cultures of the rat gingival epithelium. METHODS: The gingival epithelium was obtained from Sprague-Dawley rats and was cultured in serum-free keratinocyte growth medium (DK-SFM). Western blotting analysis, immunofluorescence, confocal laser scanning microscopy (CLSM), and immuno-gold labeling for laminin gamma(2) and integrin beta(4) were employed. CLSM images for laminin and integrin were analyzed in horizontal (x-y axis) and in vertical (x-z axis) sections. RESULTS: Both laminin gamma(2) and integrin beta(4) were detected by Western blot analysis in the gingival epithelium. Immunolocalization of laminin gamma(2) was distinct in the cytoplasm to form one or two irregular rings in gingival epithelial cells. By contrast, integrin beta(4) was localized diffusely in the cytoplasm. F-actin (indicating actin filaments) was clearly discernible at the periphery of the cytoplasm to form a cellular fringe. In x-z axis images obtained by CLSM, laminin gamma(2) was recognized as large foci in the most inner portion just above the basal plasma membrane. Integrin beta(4) existed in the area where F-actin was labeled surrounding the membrane. Immuno-electron microscopy showed that 10nm colloidal gold particles indicating laminin gamma(2) were mainly localized at the extracellular portion and in the peripheral cytoplasm, whereas integrin beta(4) was distributed in the cytoplasm close to the basal plasma membrane but not in extracellular regions. CONCLUSIONS: In primary cultures of the rat gingival epithelium, both laminin gamma(2) and integrin beta(4) may be produced by the epithelium, and irregular rings of laminin gamma(2) are formed in areas where gingival cells adhere to the extracellular matrix.     

成人牙周炎龈上皮基底膜改变的研究

应用层粘连蛋白和Ⅳ型胶原的单克隆抗体对中度至重度成人慢生牙周炎龈上皮基底膜进行免疫组化研究，并结合电镜和ＰＡＳ技术加以分析。结果发现，正常时龈上皮基底膜呈连续均匀的带状分布，而炎症时则失去正常外观，出现多个散在的微小断裂，甚至个别上皮钉突基 膜缺失     

Integrin α3β1 as a breast cancer target

Introduction: Integrin receptors for cell adhesion to the extracellular matrix have important roles in all stages of cancer progression and metastasis. Since the integrin family was discovered in the early 1980's, many studies have identified critical adhesion and signaling functions for integrins expressed on tumor cells, endothelial cells and other cell types of the tumor microenvironment, in controlling proliferation, survival, migration and angiogenesis. In recent years, the laminin-binding integrin α3β1 has emerged as a potentially promising anti-cancer target on breast cancer cells.

Areas covered: Studies from the past decade that implicate integrins as promising anti-cancer targets and the development of integrin antagonists as anti-cancer therapeutics. Recent preclinical studies that have identified the laminin-binding integrin α3β1 as an appealing anti-cancer target and the knowledge gaps that must be closed to fully exploit this integrin as a therapeutic target for breast cancer.

Expert opinion: Although the tumor-promoting functions of α3β1 implicate this integrin as a promising therapeutic target on breast cancer cells, successful exploitation of this integrin as an anti-cancer target will require a better understanding of the molecular mechanisms whereby it regulates specific tumor cell behaviors and the identification of the most appropriate α3β1 functions to antagonize on breast cancer cells.     

Amino‐terminal fragments of laminin γ2 chain retract vascular endothelial cells and increase vascular permeability

Laminin γ2 (Lmγ2) chain, a subunit of laminin‐332, is a typical molecular marker of invading cancer cells, and its expression correlates with poor prognosis of cancer patients. It was previously found that forced expression of Lmγ2 in cancer cells promotes their invasive growth in nude mice. However, the mechanism of the tumor‐promoting activity of Lmγ2 remains unknown. Here we investigated the interaction between Lmγ2 and vascular endothelial cells. When treated with an N‐terminal proteolytic fragment of γ2 (γ2pf), HUVECs became markedly retracted or shrunken. The overexpression of Lmγ2 or treatment with γ2pf stimulated T‐24 bladder carcinoma cells to invade into the HUVEC monolayer and enhanced their transendothelial migration in vitro. Moreover, γ2pf increased endothelial permeability in vitro and in vivo. As the possible mechanisms, γ2pf activated ERK and p38 MAPK but inactivated Akt in HUVECs. Such effects of γ2pf led to prominent actin stress fiber formation in HUVECs, which was blocked by a ROCK inhibitor. In addition, γ2pf induced delocalization of VE‐cadherin and β‐catenin from the intercellular junction. As possible receptors, γ2pf interacted with heparan sulfate proteoglycans on the surface of HUVECs. Moreover, we localized the active site of γ2pf to the N‐terminal epidermal growth factor‐like repeat. These data suggest that the interaction between γ2pf and heparan sulfate proteoglycans induces cytoskeletal changes of endothelial cells, leading to the loss of endothelial barrier function and the enhanced transendothelial migration of cancer cells. These activities of Lmγ2 seem to support the aberrant growth of cancer cells.     

Extracellular matrix protein expression in cerebrospinal fluid from patients with tropical spastic paraparesis associated with HTLV‐I and Creutzfeldt‐Jakob disease

The cerebrospinal fluid (CSF) is in direct contact with the extracellular space of the CNS, thus biochemical processes in the CNS could potentially be reflected in the CSF. Changes in extracellular matrix (ECM) proteins can be studied through their analysis in the CSF. ECM plays an essential role in CNS homeostasis and several proteins such as laminin (LN), fibronectin (FN), thrombospondin (TS) and heparan sulphate proteoglycan (HS, perlecan) form part of its structure. Possible changes in the levels of these proteins were investigated in two different pathologies —tropical spastic paraparesis/HTLV‐I‐associated myelopathy (TSP/HAM) (n=25) and Creutzfeldt‐Jakob disease (CJD) (n=19)—and compared with those in a control group with or without neurological disease (n=25). CSF analyses were carried out using monoclonal or monospecific polyclonal antibodies. In comparison with the control group, it was found that TSP/HAM patients presented significantly higher levels of LN, TS and HS, while in CJD patients the levels of FN, TS and HS were increased. In CJD patients the HS level was almost double that of the TSP/HAM patients. These results suggest a distinct pattern of ECM proteins in CSF in relation to the type of neurological disease. TSP/HAM is a chronic motor disease that affects the white matter of the spinal cord, while CJD is a subacute dementia that affects cerebral neurons and their synapsis.     

联合检测慢性肾功能衰竭患者血清肝纤维化指标及Cys C水平的临床应用

目的探讨联合检测血清肝纤维化指标与血清胱抑素(Cys C)在慢性肾功能衰竭(CRF)患者中的临床应用价值。方法 CRF组55例,健康对照组32例,采用放射免疫分析法检测血清透明质酸(HA)、Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(C-Ⅳ)层黏连蛋白(LN)水平,同时采用全自动生化仪检测血清Cys C、尿素氮(BUN)、肌酐(Cr)。结果 CRF组患者HA、PC-Ⅲ、C-Ⅳ、LN分别为(316.47±48.69)、(258.42±49.74)、(110.79±22.45)、(142.11±28.16)ng/mL,血清Cys C为(3.83±1.28)ng/mL。健康对照组HA、PC-Ⅲ、C-Ⅳ、LN水平则分别为(85.43±20.68)、(76.52±40.12)、(62.28±18.72)和(106.08±27.13)ng/mL,血清Cys C为(0.86±0.18)mg/mL,除LN外,其他指标CRF组明显高于健康对照组,差异有统计学意义(P<0.01)。结论联合检测血清肝纤维化指标及血清Cys C比单项检测对于CRF患者的早期诊断及疗效观察更有临床意义。     

LAMA2 gene analysis in a cohort of 26 congenital muscular dystrophy patients

Congenital muscular dystrophy type 1A (MDC1A) is caused by mutations in the LAMA2 gene encoding laminin-alpha2. We describe the molecular study of 26 patients with clinical presentation, magnetic resonance imaging and/or laminin-alpha2 expression in muscle, compatible with MDC1A. The combination of full genomic sequencing and complementary DNA analysis led to the particularly high mutation detection rate of 96% (50/52 disease alleles). Besides 22 undocumented polymorphisms, 18 different mutations were identified in the course of this work, 14 of which were novel. In particular, we describe the first fully characterized gross deletion in the LAMA2 gene, encompassing exon 56 (c.7750-1713_7899-2153del), detected in 31% of the patients. The only two missense mutations detected were found in heterozygosity with nonsense or truncating mutations in the two patients with the milder clinical presentation and a partial reduction in muscle laminin-alpha2. Our results corroborate the previous few genotype/phenotype correlations in MDC1A and illustrate the importance of screening for gross rearrangements in the LAMA2 gene, which may be underestimated in the literature.     

Expression and arrangement of extracellular matrix proteins in the lungs of mice infected with Paracoccidioides brasiliensis conidia

Extracellular matrix (ECM) proteins are important modulators of migration, differentiation and proliferation for the various cell types present in the lungs; they influence the immune response as well as participate in the adherence of several fungi including Paracoccidioides brasiliensis. The expression, deposition and arrangement of ECM proteins such as laminin, fibronectin, fibrinogen, collagen and proteoglycans in the lungs of mice infected with P. brasiliensis conidia has been evaluated in this study, together with the elastic fibre system. Lungs of BALB/c mice infected with P. brasiliensis conidia were analysed for the different ECM proteins by histological and immunohistochemical procedures at different times of infection. In addition, laser scanning confocal microscopy and scanning electron microscopy were used. During the early periods, the lungs of infected animals showed an inflammatory infiltrate composed mainly of polymorphonuclear neutrophils (PMNs) and macrophages, while during the later periods, mice presented a chronic inflammatory response with granuloma formation. Re-arrangement and increased expression of all ECM proteins tested were observed throughout all studied periods, especially during the occurrence of inflammatory infiltration and formation of the granuloma. The elastic fibre system showed an elastolysis process in all experiments. In conclusion, this study provides new details of pulmonary ECM distribution during the course of paracoccidioidomycosis.     

Laminin and integrin expression in the ventral ectodermal ridge of the mouse embryo: Implications for regulation of BMP signalling

Background: The ventral ectodermal ridge (VER) is an important signalling centre in the mouse tail‐bud following completion of gastrulation. BMP regulation is essential for VER function, but how these signals are transmitted between adjacent tissues is unclear. Results: We investigated the idea that extracellular matrix components might be involved, using immunohistochemistry and in situ hybridisation to detect all known α, β, and γ laminin chains and their mRNAs in the early tail bud. We identified an apparently novel laminin variant, comprising α5, β3 and γ2 chains, as a major component of the VER basement membrane at E9.5. Strikingly, only the mRNAs for these chains were co‐expressed in VER cells, suggesting that lamin532 may be the sole basement membrane laminin at this stage. Since α6 integrin was also expressed in VER cells, this raises the possibility of cell‐matrix interactions regulating BMP signalling at this site of caudal morphogenesis. Conclusions: Laminin532 could interact with α6‐containing integrin to direct differentiation of the specialised VER cells from surface ectoderm. Developmental Dynamics 241:1808–1815, 2012. © 2012 Wiley Periodicals Inc.