Distinct role of MAPKAPK-2 in the regulation of TNF gene expression by Toll-like receptor 7 and 9 ligands

Toll-like receptor ligands (TLRLs) produced by various pathogens activate mitogen-activated protein kinases (MAPKs). While the dependence on p38 MAPK activation for the induction of inflammatory genes by the TLR4L, lipopolysaccharide (LPS), has been well documented, the importance of the p38 pathway in gene regulation by other TLRLs is less well understood. We have focused our analysis on two TLRLs with therapeutic potential, imidazoquinoline S28463 (TLR7L) and CpG DNA (TLR9L), to explore in detail their effects on the regulation of gene expression in macrophages. Here we report that activation of the p38 MAPK/MK2 pathway is crucial for both S28463- and CpG-induced cytokine and chemokine production. We show that the stability of TNF mRNA induced by CpG DNA and S28463 is not dependent on the p38 MAPK/MK2 pathway, in contrast to LPS-induced TNF mRNA. Using a GFP reporter construct under the control of the 3' untranslated region of the TNF gene, we demonstrate that S28463 and CpG DNA-induced MK2 signalling regulates TNF mRNA primarily at the translational level, whereas LPS-induced MK2 signalling regulates both the stability and translational efficiency of TNF mRNA. Overall, these data provide insight into distinct molecular mechanisms of gene expression regulation by different Toll-like receptor ligands.     

非小细胞肺癌表皮生长因子受体突变的检测及临床病理意义

目的检测非小细胞肺癌患者体细胞表皮生长因子受体（EGFR）基因第19和21外显子突变情况并探讨其临床病理联系。方法用酚．氯仿法抽提66例NSCLC患者手术标本的基因组DNA,采用PCR技术扩增EGFR基因第19和21号外显子,从正反两个方向对扩增片段进行DNA测序和分析,并寻找EGFR突变与患者115床病理特征之间的联系。结果66例NSCLC患者中有11例（16．7％）存在EGFR杂合性体细胞突变,其中7例为第19外显子缺失突变,4例为第21外显子替代突变。女性患者突变率（9／34,26,5％）高于男性患者突变率（2／32,6．3％）,腺癌患者突变率（10／43,23,3％）高于鳞癌（0／13）和腺鳞癌患者（1／10）,吸烟者与非吸烟者突变率之间差异无统计学意义。伴细支气管肺泡癌成分的腺癌患者EGFR突变率（6／11）,高于无细支气管肺泡癌成分的腺癌患者（4／32,12．5％）。结论EGFR突变率以伴细支气管肺泡癌成分的腺癌和女性较高,更有利于适宜靶向治疗患者的临床筛选。     

Heme oxygenase-1 attenuates ovalbumin-induced airway inflammation by up-regulation of foxp3 T-regulatory cells, interleukin-10, and membrane-bound transforming growth factor- 1

Cumulative evidence suggests the up-regulation of interleukin (IL)-10 and T-regulatory (Treg) cells is implicated in anti-inflammatory effect of heme oxygenase-1 (HO-1). Thus, we postulated that induction of HO-1 could augment IL-10 and transforming growth factor (TGF)-beta production and foxp3+CD4+CD25+ Treg cell function, thereby leading to attenuation of airway inflammation. In this study, CD4+CD25+ Treg cells isolated from mouse spleen were either transfected with a HO-1 expression vector (pcDNA3HO-1) or treated with a HO-1 inducer (hemin). Up-regulation of HO-1 enhanced foxp3 expression and IL-10 secretion in the Treg cells in vitro. Next, BALB/c, C57/B6.129, and IL-10-deficient B6.129P2-Il10tm1Cgn/J mice were challenged by ovalbumin to induce airway inflammation. Consistent with in vitro findings, hemin treatment resulted in induction of HO-1 and foxp3 and production of IL-10 and membrane-bound TGF-beta1 in vivo. This was further correlated with decrease of ovalbumin-specific immunoglobulin E level and eosinophil infiltration in bronchial alveolar lavage fluid from the asthmatic mice. Furthermore, hemin significantly enhanced the biological activity of CD4+CD25+ Treg cells. This protective effect was specifically blocked by Sn-protoporphyrin, a HO-1 enzymatic inhibitor. Finally, hemin failed to up-regulate the function of CD4+CD25+ Treg cells from IL-10-deficient mice. Our study indicates that HO-1 exerts its protective effect on asthma through a mechanism mediated by foxp3+CD4+CD25+ Treg cells, IL-10, and membrane-bound TGF-beta1.     

Resveratrol relieves particulate matter (mean diameter < 2.5 μm)‐induced oxidative injury of lung cells through attenuation of autophagy deregulation

Oxidative stress and inflammation are critically implicated in ambient fine particulate matter (mean diameter < 2.5 μm; PM_2.5)‐induced lung injury. Autophagy, playing a crucial role in various physiopathological conditions, modulates cellular homeostasis and stress adaptation. Resveratrol is a phytoalexin that exerts potent antioxidant effects on cardiopulmonary diseases. To date, the mechanisms by which resveratrol protects against PM_2.5 remain to be elucidated. In the present study, we investigated the effect of resveratrol on PM_2.5‐induced oxidative injury. The potential role of nuclear factor erythroid‐2‐related factor 2 and autophagy in this progress was explored. Human bronchial epithelial cells were treated with PM_2.5 and the cytotoxicity and oxidative stress markers were determined. The results showed that PM_2.5 decreased cell viability and elevated the level of lactate dehydrogenase. The levels of malondialdehyde and reactive oxygen species were increased by PM_2.5 exposure. PM_2.5 also induced a significant increase of the inflammatory cytokines including interleukin (IL)‐6, IL‐8, IL‐1β and tumor necrosis factor α. Meanwhile, PM_2.5 triggered autophagy formation and alteration of the nuclear factor erythroid‐2‐related factor 2 pathway. Furthermore, human bronchial epithelial cells were co‐treated with PM_2.5 and resveratrol in the presence or absence of 3‐methylamphetamine, an inhibitor of autophagic formation. It was revealed that resveratrol intervention abolished PM_2.5‐induced oxidative injury partially through the suppression of autophagy deregulation. Findings from this study could provide new insights into the molecular mechanisms of pulmonary intervention during PM_2.5 exposure.     

农村地区培养卫生人才的国际经验与启示

目的:描述国外为农村地区培养卫生人才的政策和项目,分析增加农村地区卫生人力的做法和措施,为我国订单定向免费医学生培养政策提供参考和依据。方法:采用文献综述方法,搜索国内外为农村地区培养卫生人才相关文献48篇,对上述文献进行逻辑分析归纳,系统回顾分析国内外相关策略和做法,总结经验启示。结果:为农村地区培养卫生人才的常见措施包括:招收农村背景学生、导师制医学培养、农村卫生机构实习、经济激励措施、毕业后强制农村服务。对我国的启示包括:充足的财政资金是保证项目顺利实施的先决条件;做好项目管理和评价工作可以提高项目实施效果;选择合适的做法组合实施此类项目可以提高其效果。结论:订单定向免费医学生培养政策适合当前我国国情,同时不少国外经验值得我们借鉴:加大资金投入,加强管理和评价,加强部门合作,进一步完善订单定向医学生培养。     

A-40926混合物中B0组分的分离纯化

目的 开发一种高纯度A-40926混合物中B0组分(以下简称A-40926 B0)的分离纯化工艺。方法 采用中压色谱系统,以聚合物微球为载体,A-40926 B0的收率为指标,筛选聚合物微球的类型,优化洗脱条件。结果 型号为UniPS30-300的微球为最佳层析介质,上样量为10mg(每毫升填料),洗脱剂依次为体积分数30%,45%的乙醇溶液,流速为1.5BV/h(BV：层析柱体积);所得产品纯度大于95%,层析总收率大于75%。结论 此种高纯度A-40926 B0的分离纯化方法简单易行,收率稳定,适于工业化生产。     

PDCA循环管理法在抗菌药物管理中的应用

目的：评价PDCA循环法在改善医院抗菌药物临床应用中的效果。方法：采用PDCA循环管理法,通过计划-执行-检查-改进环节对武汉大学中南医院抗菌药物的临床应用进行专项整治,并对整治活动前后（2012~2016年）的抗菌药物使用强度、住院患者抗菌药物使用率以及抗菌药物使用前病原微生物送检率的效果进行分析、评价。结果：经过PDCA循环干预,我院抗菌药物的临床应用有了显著改善,抗菌药物使用率由73.37%降至38.64%,抗菌药物使用强度由68.43降至29.02,限制级和特殊使用级抗菌药物的病原微生物送检率由21.59%和57.34%分别上升至88.42%和99.07%,I类切口预防使用抗菌药物比例由84.5%降至28.6%。结论：PDCA循环法用于医院抗菌药物的管理可促进抗菌药物的合理使用,提高医院合理用药水平。     

New resilience instrument for patients with cancer

Objective

Resilience is an important concept in the cancer literature and is a salient indicator of cancer survivorship. The aim of this study is to develop and validate a new resilience instrument that is specific to patients with cancer diagnosis (RS-SC) in Mainland China.

Methods

First, a resilience framework was constructed for patients with cancer diagnosis. Second, items were formulated based on the framework to reflect different aspects of resilience. Third, two rounds of expert panel discussion were performed to select important and relevant items. Finally, two cross-sectional studies were conducted to evaluate the psychometric properties of this instrument.

Results

Fifty-one items were generated based on the resilience framework and the final 25-item RS-SC resulted in a five-factor solution including Generic Elements, Benefit Finding, Support and Coping, Hope for the Future and Meaning for Existence, accounting for 64.72% of the variance. The Cronbach’s α of the RS-SC was 0.825 and the test–retest reliability was 0.874.

Conclusion

The RS-SC is a brief and specific self-report resilience instrument for Chinese patients with cancer and shows sound psychometric properties in this study. The RS-SC has potential applications in both clinical practice and research with strength-based resiliency interventions.

     

Targeted delivery of hyaluronic acid-coated solid lipid nanoparticles for rheumatoid arthritis therapy

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disease. Long-term, high-dose glucocorticoid therapy can be used to treat the disease, but the fact that the drug distributes systemically can give rise to severe adverse effects. Here we develop a targeted system for treating RA in which the glucocorticoid prednisolone (PD) is encapsulated within solid lipid nanoparticles (SLNs) coated with hyaluronic acid (HA), giving rise to HA-SLNs/PD. HA binds to hyaluronic receptor CD44, which is over-expressed on the surface of synovial lymphocytes, macrophages and fibroblasts in inflamed joints in RA. As predicted, HA-SLNs/PD particles accumulated in affected joint tissue after intravenous injection into mice with collagen-induced arthritis (CIA), and HA-SLNs/PD persisted longer in circulation and preserved bone and cartilage better than free drug or drug encapsulated in SLNs without HA. HA-SLNs/PD reduced joint swelling, bone erosion and levels of inflammatory cytokines in serum. These results suggest that encapsulating glucocorticoids such as PD in HA-coated SLNs may render them safe and effective for treating inflammatory disorders.     

The anti-tumor effect of pachymic acid on osteosarcoma cells by inducing PTEN and Caspase 3/7-dependent apoptosis

Pachymic acid (PA) is a lanostane type triterpenoid isolated from Poria cocos, which possesses an anti-tumor effect in breast cancer, prostate cancer, lung cancer, and bladder cancer cells. In this study, we investigated the effect of PA on the growth and apoptosis of human immortalized cell line (HOS) and primary osteosarcoma cells by a Cell Counting Kit-8 (CCK-8) and Annexin V and propidium iodide (PI) staining, respectively. Western blot was used to measure the expression of cleaved Caspase 3, PTEN, and AKT, as well as the AKT phosphorylation. The Caspase 3 activity was determined using the Caspase-3 Colorimetric Assay Kit. From the results, PA significantly reduced cell proliferation in a concentration- and time-dependent manner. PA also induced cell apoptosis in a dose-dependent fashion. PA treatment led to increased Caspase 3 activation and PTEN expression, as well as reduced AKT phosphorylation. Moreover, Ac-DEVD-CHO (a Caspase 3/7 inhibitor) pre-treatment or PTEN knockdown partially blocked the effects of PA on cell proliferation and apoptosis. Caspase 3/7 inhibitor had an additive effect with PTEN knockdown. Collectively, our results suggested that induction of apoptosis by PA was mediated in part by PTEN/AKT signaling and Caspase 3/7 activity. This study provides evidence that PA might be useful in the treatment of human osteosarcoma.