A method for measuring mouse respiratory allergic reaction to low-dose chemical exposure to allergens: An environmental chemical of uncertain allergenicity,a typical contact allergen and a non-sensitizing irritant

Our aim was to improve a method for detecting respiratory hypersensitivity by testing three confirmed respiratory allergens (trimellitic anhydride [TMA], phthalic anhydride [PA] and toluene diisocyanate [TDI]), an environmental chemical of uncertain allergenicity (2,4-d-butyl [DB]), a confirmed contact allergen (2,4-dinitrochlorobenzene [DNCB]) and a standard irritant (sodium dodecyl sulfate [SDS]). BALB/c mice were topically sensitized (nine times in 3 weeks) with these chemicals, then challenged via the trachea. One day post-challenge, samples were taken from the mice to assay for total immunoglobulin (IgE and IgG₁) levels in serum and bronchoalveolar lavage fluid (BALF); differential cell counts and cytokine/chemokine levels in BALF; lymphocyte counts and surface antigen expression on B-cells within lung-associated lymph nodes (LNs); ex situ cytokine production by cells from these LNs; and gene expression in BALF (CCR3) and LNs (CCR4, STAT6 and GATA-3). The three confirmed respiratory allergens and DB induced immune response characteristic of immediate-type respiratory reactions, as evidenced by increased total IgE and IgG₁ levels; influx of eosinophils, neutrophils, chemokines and cytokines in BALF; increased surface antigen expression on B-cells in LNs; increased Th2 cytokine production in LNs; and increased respiratory allergy-related gene expression in both BALF and LNs. In contrast, DNCB and SDS treatments yielded, at most, insignificant increases in all respiratory allergic parameters. Thus, the protocol was equally suitable for use with an environmental chemical of unknown allergenicity and for typical respiratory allergens. Since the protocol differentiated respiratory allergens from contact allergens and irritants, it may be useful for detecting respiratory allergy related to environmental chemicals.     

Overexpression of interleukin-12 and T helper 1 predominance in lupus nephritis

Imbalance of cytokine homeostasis is a prominent feature of both experimental and human systemic lupus erythematosus (SLE). Because interleukin (IL)-12 promotes interferon (IFN)-gamma production leading to polarization of peripheral cells toward a T helper (Th) 1 phenotype, we investigated its role in lupus nephritis (LN). Soluble Th1 and Th2 cytokines were measured by enzyme-linked immunosorbent assay (ELISA) in sera and urines of SLE patients and controls. Th1/Th2 peripheral lymphocyte polarization was determined by flow cytometry. Glomerular accumulation of IL-12 was evaluated by immunohistochemistry, whereas urinary IL-12 was evaluated by ELISA. Higher serum IL-12 levels in SLE were associated with LN, whereas IL-4 was unrelated to the renal damage. Peripheral cells from LN patients showed a Th1 phenotype with a high IFN-gamma expression that paralleled the severity of renal damage. IL-12 was present within glomerular mononuclear cells in classes IV and V LN, and its accumulation was correlated strongly with urinary levels. IL-12 overexpression in SLE may contribute to the development of LN. Both serum and urinary IL-12 elevation reflect its glomerular production and parallel Th1 polarization of peripheral T cells and high IFN-gamma production. In SLE patients, IL-12 measurement may thus be predictive of the development of LN.     

Single stage immediate breast reconstruction with acellular dermal matrix and implant: Defining the risks and outcomes of post-mastectomy radiotherapy

IntroductionThe objective of this study is to evaluate outcomes and complications in patients with single-stage ADM-implant based immediate breast reconstruction with and without radiotherapy (RT), highlighting the effects of RT on the reconstruction.Materials and methodsThis prospective study recruited 91 consecutive patients who underwent skin-sparing, nipple-sparing or wise-pattern skin reduction mastectomy with direct-to-implant breast reconstruction with ADMs using sub-pectoral or pre-pectoral approach at the two breast units. Early and late complications like seroma, delayed wound healing, wound breakdown, infection, capsular contracture, implant loss and revision surgery were evaluated in the RT and non-RT groups.ResultsIn the total cohort of 91 patients, 29 received adjuvant RT and 62 did not need RT. In the RT group, 3–7% of them had early complications like seroma, wound infections and delayed healing. 20.7% had post-RT capsular contractures which either required revision surgery with autologous flap (6.9%) or capsulotomy with exchange of implant (6.9%). In the non-RT group, 7–9% cases had seroma & wound infections, 3.06% had delayed wound healing and 7.25% had capsular contracture. 13.04% required revision surgery due to infection, implant loss or failure to achieve expectations. The total loss of implants in the cohort was 7.14% (RT group 6.9% and non-RT group 7.25%). The need for PMRT could have been predicted pre-operatively in the RT group in 55.17% cases based on the extent of disease, multifocality, tumour grade and positive LN status on imaging.ConclusionADM based reconstruction in patients anticipated to receive adjuvant RT is always debatable. Though there is no significant difference in the revision surgeries in our study of the 2 groups, the rate of capsular contracture as expected, was higher in the RT group. Hence, pre-operative discussion on the need for RT highlighting the risks and complications will help patients make a better-informed choice.     

肺腺癌中LN 5蛋白表达及其与EGFR突变关系的临床研究

目的 探讨肺腺癌组织中层粘连蛋白 5（LN-5）表达及其与人表皮生长因子受体（EGFR）基因突变关系。方法 收集2015年1月至2015年12月57例肺腺癌及其配对癌旁组织标本,采用免疫组化SP法检测LN 5蛋白表达,同时通过液相芯片技术检测EGFR基因突变状态,分析LN 5表达与肺腺癌临床病理特征的关系以及与EGFR基因突变的相关性。 结果肺腺癌组织中LN 5蛋白阳性表达率为21.1％（12／57）,癌旁组织中为1.7％（1／57）,差异有统计学意义（P＜0.05）。EGFR敏感突变（E19+E21）的发生率为43.9％（25／57）,EGFR非敏感突变（野生型+E20）的发生率为56.1％（32／57）。LN-5蛋白表达与淋巴结转移和EGFR突变有关（P＜0.05）,与TNM分期无关（P＞0.05）。LN 5表达与EGFR突变呈负相关（r=-0.283,P＜0.05）。结论 LN 5在肺腺癌组织中的表达升高,且其表达与EGFR突变状态具有相关性。     

Management of positive margins after initial lumpectomy in elderly women with breast cancer

Introduction

It is unknown whether the treatment disparity observed between young and elderly women extends to the management of positive margins after initial lumpectomy. The primary aim was to evaluate the management of positive margins after initial lumpectomy in elderly women.

SENP1、SENP2和SENP6蛋白在人恶性胶质瘤组织和细胞中的表达及其意义

目的：观察SUMO特异性蛋白酶（SENPs）家族成员SENP1、SENP2和SENP6在人恶性胶质瘤组织和细胞中的表达,阐明其在恶性胶质瘤发生中的作用。方法：取人脑正常组织和恶性胶质瘤组织样品分别作为正常对照组和恶性胶质瘤组;培养Cos7细胞和恶性胶质瘤LN443和U343细胞,Cos7细胞作为正常细胞对照组,LN443和U343作为恶性胶质瘤细胞组;采用Western blotting法检测SENP1、SENP2和SENP6蛋白在人恶性胶质瘤组织和细胞中的表达水平。结果：在脑组织中,与正常对照组比较,恶性胶质瘤组恶性胶质瘤组织中SENP1、SENP2和SENP6蛋白表达水平均明显升高（P<0.05）。在细胞中,与正常细胞对照组比较,恶性胶质瘤细胞组LN443和U343细胞中SENP1、SENP2和SENP6蛋白表达水平均明显升高（P<0.05）。结论：SENP1、SENP2和SENP6在恶性胶质瘤组织和细胞中高表达,其可能对恶性胶质瘤的发生起到了重要的促进作用。     

Impact of margin status and lymphadenectomy on clinical outcomes in resected pancreatic adenocarcinoma: implications for adjuvant radiotherapy

Background

Adjuvant chemoradiotherapy (CRT) in the treatment of pancreatic ductal adenocarcinoma (PDA) is controversial. Minimal data exists regarding the clinical significance of margin clearance distance and lymph node (LN) parameters, such as extent of dissection and LN ratio. We assessed the impact of these variables on clinical outcomes to more clearly define the subset of patients who may benefit from adjuvant radiotherapy (RT).

Methods

We identified 106 patients with resected stage 1-3 PDA from 2007-2013. Resection margins were categorized as positive (tumor at ink), ≤1, or >1 mm. LN evaluation included total number examined (NE), number of positive nodes (NP), ratio of NP to NE (NR), extent of dissection, and positive periportal LNs. The impact of these variables was assessed on disease-free survival (DFS) and overall survival (OS) using multivariate cox proportional hazards modeling.

Results

In patients receiving adjuvant chemotherapy (CT) alone, greater margin clearance led to improved DFS (P=0.0412, HR =0.51). Range of NE was 4-37, with a mean of 19. NE was not associated with DFS or OS, yet absolute NP of 5 or more was associated with a significantly worse DFS (P=0.005). Whereas periportal lymphadenectomy did not result in improved DFS or OS, patients with positive periportal LN had worse clinical outcomes (DFS, P=0.0052; OS, P=0.023). The use of adjuvant CRT was associated with improved OS (P=0.049; HR=0.29).

Conclusions

In patients receiving adjuvant CT alone, there was a clinically significant benefit to clearing the surgical margin beyond tumor at ink. Having ≥5 NP and positive periportal LN led to significantly worse clinical outcomes. The addition of adjuvant RT to CT in resected PDA improved OS. A comprehensive evaluation of resection margin distance and LN parameters may identify more patients at risk for locoregional failure who may benefit from adjuvant CRT.     

miR‐410 suppresses the expression of interleukin‐6 as well as renal fibrosis in the pathogenesis of lupus nephritis

Lupus nephritis (LN) is a highly complex autoimmune disease caused by systemic lupus erythematosus (SLE). MicroRNAs (miRNAs) play a vital role in the pathogenesis of SLE. Previously, a total of 29 miRNAs were identified to be down‐regulated in SLE patients, in which miR‐410 was likely to be involved in the signalling transduction pathways in regulating the pathogenesis of SLE. The purpose of this study was to investigate the role of miR‐410 in LN and to find out whether miR‐410 regulates the expression of interleukin (IL)‐6 and fibrosis in LN. It was found that the expression level of miR‐410 in kidney tissue of MRL/lpr mice was decreased compared to that in BALB/C mice, whereas the level of IL‐6 was overexpressed in MRL/lpr mice. Luciferase assay showed that miR‐410 binds directly to the 3′ untranslated region (UTR) of IL‐6, with the results showing that overexpression of miR‐410 significantly decreased the expression level of IL‐6 in SV40MES13 cells. Moreover, overexpression of miR‐410 significantly reduced the expression levels of fibrosis factors such as transforming growth factor‐β1 (TGF‐β1) and collagen I/III in SV40MES13 cells; Inhibition of the expression of miR‐410 with miR‐410 inhibitor resulted in increased levels of IL‐6 as well as fibrosis factors. The results identify that miR‐410, as a novel and critical factor in the pathogenesis of LN, decreases IL‐6 expression by binding directly to the 3′UTR and suppresses fibrosis through down‐regulation of TGF‐β1 in SV40MES13 cells. Our study brings new insight into understanding the complex mechanisms involved in the pathogenesis of lupus disease.     

狼疮肾炎患者循环miRNA表达谱研究及循环miR-130b-3p的临床意义

目的：研究探讨狼疮肾炎患者外周循环miRNA表达变化及其可能存在的临床意义,为狼疮肾炎的临床治疗提供更多的客观依据。方法方便选取2013年9月-2015年9月于该院诊治的50例狼疮肾炎患者,同时另选同期于该院体检的50名健康者作为对照组。在100例研究对象中随机选取15例患者做循环miRNA的表达谱检测筛选(检测组),85例患者做循环miRNA的表达谱验证筛选(验证组),同时应用循环miR-130b-3p为验证对象,观察与比较狼疮肾炎患者与对照组患者的血清总RNA水平及检测组中两组患者的血清miRNA水平、验证组中miR-130b-3p的水平,并进行数据统计学分析及相关性分析。结果在验证组中,与对照组相比较(3.7±1.6)ng/μL,狼疮肾炎组患者的循环血清中miR-130b-3p的表达水平(9.8±1.4)ng/μL显著上升,其差异有统计学意义(P<0.05);在检测组中,与对照组相比较(28.6±2.4)ng/μL,狼疮肾炎组患者循环表达的miRNA水平(46.8±5.6)ng/μL明显上升,其差异具有统计学意义(P<0.05)。结论狼疮肾炎患者循环miRNA表达水平可能随着肾功能的进一步下降而出现表达下调,其循环miR-130b-3p的表达与肾脏功能、血脂水平呈现明显的相关性,这在狼疮肾炎的疾病发生发展中可能起着重要的作用。