Monocyte chemotactic protein-inducing protein 1 negatively regulating asthmatic airway inflammation and mucus hypersecretion involving γ-aminobutyric acid type A receptor signaling pathway in vivo and in vitro

BackgroundMounting evidence, consistent with our previous study, showed that γ-aminobutyric acid type A receptor (GABAAR) played an indispensable role in airway inflammation and mucus hypersecretion in asthma. Monocyte chemotactic protein-inducing protein 1 (MCPIP1) was a key negative regulator of inflammation. Recent studies showed that inflammation was largely suppressed by enhanced MCPIP1 expression in many inflammatory diseases. However, the role and potential mechanism of MCPIP1 in airway inflammation and mucus hypersecretion in asthma were still not well studied. This study was to explore the role of MCPIP1 in asthmatic airway inflammation and mucus hypersecretion in both mice and BEAS-2B cells, and its potential mechanism.MethodsIn vivo, mice were sensitized and challenged by ovalbumin (OVA) to induce asthma. Airway inflammation and mucus secretion were analyzed. In vitro, BEAS-2B cells were chosen. Interleukin (IL)-13 was used to stimulate inflammation and mucus hypersecretion in cells. MCPIP1 Lentiviral vector (LA-MCPIP1) and plasmid-MCPIP1 were used to up-regulate MCPIP1 in lung and cells, respectively. MCP-1, thymic stromal lymphopoietin (TSLP), mucin 5AC (MUC5AC), MCPIP1, and GABAARβ2 expressions were measured in both lung and BEAS-2B cells. Immunofluorescence staining was performed to observe the expression of GABAARβ2 in cells.ResultsMCPIP1 was up-regulated by LA-MCPIP1 (P < 0.001) and plasmid-MCPIP1 (P < 0.001) in lung and cells, respectively. OVA-induced airway inflammation and mucus hypersecretion, OVA-enhanced MCP-1, TSLP, MUC5AC, and GABAARβ2 expressions, and OVA-reduced MCPIP1 were significantly blunted by LA-MCPIP1 in mice (all P < 0.001). IL-13-enhanced MCP-1, TSLP, MUC5AC, and GABAARβ2 expressions, and IL-13-reduced MCPIP1 were markedly abrogated by plasmid-MCPIP1 in BEAS-2B cells (all P < 0.001).ConclusionThe results of this study suggested that OVA and IL-13-induced airway inflammation and mucus hypersecretion were negatively regulated by MCPIP1 in both lung and BEAS-2B cells, involving GABAAR signaling pathway.     

川芎嗪注射液对大鼠腹膜透析效能的影响

目的 通过动物实验观察川芎嗪对腹膜透析效能的影响,并初步探讨其作用机理。方法 采用空白及实验对照设计,将３６只雄性ＳＤ大鼠平均分为单纯腹透组、多巴胺对照组、川芎嗪常规浓度组及川芎嗪高浓度组４组,观察不同剂量川芎嗪加入腹透液对大鼠尿素氮清除率（ＣＢＵＮ）、肌酐清除率（ＣＣｒ）、尿素氮Ｄ／Ｐ值、肌酐Ｄ／Ｐ值、排液量均显著高于单纯腹透组（ｐ值均小于０．０５）,３组间两两比较差异无显著性（ｐ值均大于０．０     

尼群地平片的工艺改进

将尼群地平与聚乙烯吡咯民固体分散体,再制成片剂,测定其溶出度,与普通片以及市售国产片的溶出度比较,改进片的出度明显提高,２０ｍｉｎ的累积溶出度为７０．８％,而普通片及市售国产片分别为２２．７％和２１．３％。留样观察实验结果表明,改进片质量稳定。     

ELISA法检测可溶性人类白细胞抗原—I及广东人参考值的测定

OBJECTIVE: To explore a method for quantitative detection of soluble human leukocyte antigen class I(HLA-I) antigens, with the assistance of which we attempt to define the reference value of the antigen in the population of Guangdong Province. METHODS: Sandwich enzyme-linked immunosorbent assay (ELISA) was employed with W6/32 monoclonal antibody serving as the solid phase antibody and beta2 microglobulin antibody as the first antibody. HRP-labeled second antibody and the substrate were added for enzyme-catalyzed coloring. On the basis of the standard curve obtained by sHLA-I standard reagent in serial dilution, the concentration of sHLA-I antigens in the samples of 60 normal subjects from the population of Guangdong province was detected. RESULTS: The sensitivity of this assay was 2.84 ng/ml with variation coefficients of 5.80% within the same batch and 9.00% between batches. The recovery rate ranged from 98.57% to 100.15%. The level of sHLA-I antigens in normal individuals was 699.54+/-360.10 ng/ml. CONCLUSION: Sandwich ELISA is sensitive, specific and stable in the detection of sHLA-I.     

脂质体介导人β-神经生长因子基因转染培养肌母细胞的表达研究

目的　了解含 β-神经生长因子 (β- NGF)基因的表达质粒 PSVCEP NGF- CAT在培养肌母细胞中的表达及 L ipofect AMINE阳离子脂质体介导的转染效率。方法　将表达质粒 PSVCEP NGF- CAT经 L ipofectAMINE转染至培养成的肌母细胞中 ,用免疫细胞化学方法检测基因在肌母细胞内的表达。结果　培养肌母细胞在转染 6小时吞噬脂质体最显著 ,转染 48小时后约 40 %的肌母细胞胞质内有β- NGF基因表达。结论　含β- NGF基因的表达质粒 PSVCEP NGF- CAT可以在培养肌母细胞中表达 ,脂质体转染的方法简便、有效     

麻醉诱导期三种面罩通气方式的效果比较

目的比较双人面罩手控通气（M）、面罩容量控制通气（VC）、面罩压力控制通气（PC）这三种不同的通气方式在麻醉诱导期的应用效果。方法 90例患者分为三组,M组由两名麻醉医师行双手手控面罩通气,VC组和PC组由一名麻醉医师双手扣面罩,麻醉机给予通气。VC组设定吸气潮气量（VT）8 ml/kg,呼吸频率（RR）16次/min,PC组设定RR 16次/min,调节吸气压力值以达到VT8 ml/kg。持续5min,每分钟记录呼吸、循环等参数。结果M组的呼末CO2     

MDCT与MRI诊断视网膜脱离的价值

目的探讨多排CT(multi-detectorrow CT,MDCT)和MRI对视网膜脱离的诊断价值及临床意义。方法回顾性分析2007年5月~2010年10月期间,中山大学中山眼科中心送至我院检查及中山大学附属第一医院检查的45例(47只眼)视网膜脱离患者的MDCT或MRI图像,其中16例(17只眼)经MDCT诊断,29例(30只眼)经MR诊断;45例(47只眼)视网膜脱离患者中的32例(33只眼)经手术证实,13例(14只眼)经其它临床相关检查得到验证。结果 MDCT或MRI图像上可见到脱离的视网膜与眼球后壁之间的积液征象。在所有视网膜脱离的患者中,单纯视网膜脱离21只眼;伴有其它相关征象的26只眼。伴出血者20只眼、伴钙化者6只眼。其中,脉络膜骨瘤3只眼、脉络膜黑色素瘤5只眼。结论 MDCT成像速度快、对钙化病灶显示敏感;MR无辐射,软组织分辨力高,对出血及小范围视网膜脱离显示敏感;二者对视网膜脱离均有较高的诊断价值,临床医生可根据不同患者合理选择。     

大龄腭裂患者同期腭裂修复与齿槽嵴裂植骨的临床观察

目的：探讨８岁以上单侧完全性腭裂患者同期腭裂修复与齿槽嵴裂植骨的可行性及植骨效果。方法：对３８例同期腭裂修复与齿槽嵴裂植骨的腭裂患者作回顾性研究。患者年龄８～２４岁,平均年龄为１４．８岁。分析手术时间、术中出血、术后恢复和创口愈合情况。术后随访１２月以上,对随访的Ｘ线片进行植骨效果的客观评价。结果：所有手术均顺利完成,平均手术时间比单纯改良兰氏腭裂修复手术多３７ｍｉｎ,没有明显增加术中出血量,患者     

重组人骨形成蛋白2-珊瑚复合人工骨修复骨质缺损的实验研究

目的弥补珊瑚无骨诱导活性、成骨能力弱等缺陷。方法将珊瑚与重组人骨形成蛋白２（ｒｅｃｏｍｂｉｎａｎｔｈｕｍａｎｂｏｎｅｍｏｒｐｈｏｇｅｎｅｔｉｃｐｒｏｔｅｉｎ－２，ｒｈＢＭＰ－２）复合，制备出ｒｈＢＭＰ－２－珊瑚复合人工骨。将其植入兔颅骨直径１５ｍｍ的圆形缺损，以单纯珊瑚植入作对照。术后２、６、１２周取材，通过组织学观察和计算机图像分析，评价其骨修复能力。结果ｒｈＢＭＰ－２－珊瑚复合人工骨以传导成骨和诱导成骨双重机制完成骨修复过程，骨修复能力和骨修复效果明显优于单纯的珊瑚。结论此复合人工骨是一种较理想的骨移植替代材料。     

Fluid Shear Stress Stimulates Osteogenic Differentiation of Human Periodontal Ligament Cells via the Extracellular Signal‐Regulated Kinase 1/2 and p38 Mitogen‐Activated Protein Kinase Signaling Pathways

Background: Fluid shear stress (FSS) is a major type of mechanical stress that is loaded on human periodontal ligament cells (hPDLCs) during mastication and orthodontic tooth movement. This study aims to clarify the effect of FSS on the osteogenic differentiation of hPDLCs and to further verify the involvement of mitogen‐activated protein kinase (MAPK) signaling in this process. Methods: After isolation and characterization, hPDLCs were subjected to 2‐hour FSS at 12 dynes/cm², and cell viability, osteogenic gene mRNA expression, alkaline phosphatase (ALP) activity, secretion of Type I collagen (COL‐I), and calcium deposition were assayed. The levels of phosphorylated p38 and phosphorylated extracellular signal‐regulated kinase 1/2 (ERK1/2) in response to FSS were detected by Western blot, and the involvement of ERK1/2 and p38 MAPK signaling pathways in hPDLC osteogenesis under FSS was investigated using the specific MAPK inhibitors U0126 (2Z,3Z)‐2,3‐bis[amino(2‐aminophenylthio)methylene]succinonitrile,ethanol) and SB203580 (4‐[4‐(4‐fluorophenyl)‐2‐(4‐[methylsulfinyl]phenyl)‐1H‐imidazol‐5‐yl]pyridine). Results: The application of FSS on hPDLCs induced an early morphologic change and rearrangement of filamentous actin. ALP activity, messenger RNA (mRNA) levels of osteogenic genes, COL‐I, and osteoid nodules were significantly increased by FSS. Moreover, ERK1/2 and p38 were activated in different ways after FSS exposure. U0126 and SB203580 completely blocked the FSS‐induced increases in ALP activity and osteogenic gene mRNA expression and osteoid nodules formation. Conclusions: FSS is an effective approach for stimulating osteogenic differentiation of hPDLCs. The ERK1/2 and p38 MAPK signaling pathways are involved in this cellular process.