Inter-ictal assay of peripheral circulating inflammatory mediators in migraine patients under adjunctive cervical non-invasive vagus nerve stimulation (nVNS): A proof-of-concept study

Objective

To assay peripheral inter-ictal cytokine serum levels and possible relations with non-invasive vagus nerve stimulation (nVNS) responsiveness in migraineurs.

基于“气道神经源性炎症-TRP通路”探讨慢性咳嗽发病机制及中医治疗

咳嗽临床常见，尤其是胸部影像学无明显异常的慢性咳嗽，病因复杂、易误诊误治，逐渐受到重视，国内外相继颁发相关指南，目前认为咳嗽高敏感性是其重要的病理生理机制。中医将慢性咳嗽归于"久咳""久嗽"范畴，在病因病机、治法方药上积累了丰富的临床经验，史利卿教授团队针对该病开展了专项临床及基础研究15年，根据传统中医理论及其突出临床症状提出"风邪伏肺"病机，应用祛风宣肺法治疗效果显著。文章结合西医学相关咳嗽机制研究进展及既往研究结果，初步探讨祛风宣肺法治疗该病的疗效机制，丰富慢性咳嗽风邪伏肺致咳相关学术理论的科学内涵，有利于提高中医药防治慢性咳嗽学术水平。     

Chronic inflammation and risk of lung cancer in older adults in the health,aging and body composition cohort study

Objectives

We examined the association between three inflammatory markers (Interleukin (IL)-6, C-reactive protein (CRP), tumor necrosis factor (TNF)-α) and incident lung cancer using baseline, updated, and averaged inflammatory measures in older adults.

MicroRNA-301b-3p accelerates the growth of gastric cancer cells by targeting zinc finger and BTB domain containing 4

MicroRNAs (miRNAs) have been found to be aberrantly expressed and exert essential roles in the tumorigenesis and progression of gastric cancer (GC). miR-301b-3p has been recognized as a cancer-related miRNA in lung cancer, bladder cancer and hepatocellular carcinoma. However, the function of miR-301b-3p in GC progression and its underlying mechanism have not been studied yet. In this study, we found that miR-301b-3p expression was up-regulated in GC tissues compared to adjacent noncancerous tissues. Furthermore, the elevated levels of miR-301b-3p were detected in GC cell lines (SGC-7901, AGS, MKN-45 and MGC-803) as compared with GES-1 cells. Interestingly, GC tissues from patients with tumor size ≥ 5 cm and advanced tumor stages showed obvious higher levels of miR-301b-3p compared to matched controls. Functionally, miR-301b-3p knockdown prominently inhibited cell proliferation, and induced cell cycle arrest at G1 phase and apoptosis in MGC-803 cells. Meanwhile, ectopic expression of miR-301b-3p conversely regulated these biological behaviors of MKN-45 cells. Next, we found that miR-301b-3p knockdown increased, whereas miR-301b-3p overexpression reduced the expression of zinc finger and BTB domain containing 4 (ZBTB4) in GC cells. Accordingly, luciferase reporter assay identified ZBTB4 as a direct target of miR-301b-3p. ZBTB4 overexpression markedly restrained the growth of MGC-803 cells. More importantly, ZBTB4 silencing partially reversed miR-301b-3p knockdown-induced tumor suppressive effects on MGC-803 cells. In conclusion, we firstly revealed that miR-301-3p was highly expressed in GC and contributed to tumor progression via attenuating ZBTB4, which might provide a novel molecular-targeted strategy for GC treatment.     

Detection of recombinant and endogenous mouse melatonin receptors by monoclonal antibodies targeting the C‐terminal domain

Melatonin receptors play important roles in the regulation of circadian and seasonal rhythms, sleep, retinal functions, the immune system, depression, and type 2 diabetes development. Melatonin receptors are approved drug targets for insomnia, non‐24‐hour sleep‐wake disorders, and major depressive disorders. In mammals, two melatonin receptors (MTRs) exist, MT₁ and MT₂, belonging to the G protein‐coupled receptor (GPCR) superfamily. Similar to most other GPCRs, reliable antibodies recognizing melatonin receptors proved to be difficult to obtain. Here, we describe the development of the first monoclonal antibodies (mABs) for mouse MT₁ and MT₂. Purified antibodies were extensively characterized for specific reactivity with mouse, rat, and human MT₁ and MT₂ by Western blot, immunoprecipitation, immunofluorescence, and proximity ligation assay. Several mABs were specific for either mouse MT₁ or MT₂. None of the mABs cross‐reacted with rat MTRs, and some were able to react with human MTRs. The specificity of the selected mABs was validated by immunofluorescence microscopy in three established locations (retina, suprachiasmatic nuclei, pituitary gland) for MTR expression in mice using MTR‐KO mice as control. MT₂ expression was not detected in mouse insulinoma MIN6 cells or pancreatic beta‐cells. Collectively, we report the first monoclonal antibodies recognizing recombinant and native mouse melatonin receptors that will be valuable tools for future studies.     

清热通利汤对宫颈炎并HPV感染患者生化指标与临床症状的影响研究

目的:观察清热通利汤对于宫颈炎合并人乳头瘤病毒(HPV)感染患者生化指标与临床症状的功效。方法:选取86例宫颈炎合并HPV感染患者作为观察资料,选取的患者按简单随机原则以数字表平均划分为对照组与治疗组,对照组给予常规治疗,治疗组给予清热通利汤外用; 治疗后3个月,对比临床疗效,统计临床症状的缓解时间; 评估治疗前、后患者生化指标、中医证候积分、血清炎性因子的变化情况。结果:治疗组显效率为53.49%、总有效率为95.35%,对照组依次为37.21%、72.09%,治疗组临床疗效优于对照组(P<0.05)。治疗后,治疗组患者HPV-DNA转阴率高于对照组,HPV-DNA转阴时间、HPV病毒载量均低于对照组,差异均有统计学意义(P<0.05); 治疗组患者宫颈柱状上皮异位、接触性出血、阴道清洁度异常、白带化验异常的消退时间均短于对照组(P<0.05); 治疗后两组患者各项中医证候积分、血清炎性因子与生化指标均较治疗前改善且治疗组均优于对照组(P<0.05)。结论:清热通利汤能够促进宫颈炎合并HPV感染患者HPV-DNA转阴,提高临床疗效,更有效的缓解临床症状、改善生化指标、减轻炎性反应程度。     

胶质母细胞瘤差异表达基因的生物信息学分析

目的:应用生物信息学方法挖掘胶质母细胞瘤(GBM)的相关基因，进而探讨发病机制，为GBM临床诊断和靶向治疗提供理论依据。方法:从GEO(Gene Expression Omnibus)数据库下载基因芯片数据集GSE4290和GSE15824，应用GEO2R筛选GBM的差异表达基因(DEGs)。采用DAVID数据库进行GO富集和KEGG通路富集分析，分别应用STRING数据库和Cytoscape软件构建蛋白质相互作用网络和关键基因模块，筛选GBM靶基因。进一步运用ONCOMINE数据库验证临床组织样本中靶基因与GBM的关系。结果:共筛选出76个DEGs，富集分析结果显示DEGs在血管生成的正调节、抗原的呈递和处理、信号转导、调节自噬等方面存在显著富集。共挖掘出POSTN、TAGLN、CALD1、EPCAM 4个GBM靶基因，经证实均在临床GBM组织样本中存在显著上调且靶基因的上调与患者的不良预后密切相关。结论:通过生物信息学共挖掘出4个与GBM显著相关的靶基因，可能是未来GBM发病机制、临床诊断、治疗的重要研究靶点。     

小柴胡汤对非酒精性脂肪性肝炎模型小鼠的保护作用研究

目的通过构建蛋氨酸-胆碱缺乏(MCD)饮食诱导的小鼠非酒精性脂肪性肝炎(NASH)模型,研究小柴胡汤对NASH模型小鼠的保护作用。方法选择C57BL/6小鼠为研究对象,将其随机分为对照组、模型组、小柴胡汤(高、中、低剂量)组、易善复组和强肝胶囊组。通过饲喂MCD饲料建立NASH模型,造模同时按分组给予不同药物进行干预;实验过程中记录小鼠体质量、日摄食量、日饮水量变化,实验结束对肝组织进行HE染色观察病理变化,检测血清生化指标丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、白细胞介素6(IL-6)和肿瘤坏死因子-α(TNF-α)的水平变化,检测肝组织中TC、TG的水平变化,利用q RT-PCR技术检测肝组织脂肪酸合成酶(FAS)和固醇调节元件结合蛋白1c(SREBP-1c)的表达水平。结果小鼠体质量、日摄食量、日饮水量及肝脏系数等数据显示MCD饮食诱导的模型组小鼠会出现体质量降低、摄入量减少及肝脏湿质量下降的特点,而小柴胡汤给药组小鼠体质量、摄入量及肝脏系数较模型组小鼠显著升高;HE染色结果显示小柴胡汤可明显减轻肝组织脂肪变性和炎症程度,改善肝细胞的形态和结构;生化指标检测结果显示小柴胡汤能显著降低NASH模型小鼠血清及肝组织TG、TC水平,升高血清中HDL-C水平,降低血清中AST、ALT、IL-6、TNF-α水平;q RT-PCR结果显示模型组小鼠肝组织FAS和SREBP-1c的基因表达水平明显升高,小柴胡汤可显著降低FAS和SREBP-1c的基因表达水平。结论小柴胡汤对MCD饮食诱导的NASH模型小鼠有明显保护作用,其机制可能通过调控抑制脂肪酸合成基因(FAS、SREBP-1c)的表达,减少脂肪堆积,实现调脂作用,并通过抑制炎症因子的表达改善肝组织的损伤。     

The co-expression characteristics of LAG3 and PD-1 on the T cells of patients with breast cancer reveal a new therapeutic strategy

Many studies have shown a special interaction between LAG3 and PD-1 in T cell inhibition, while the co-expression and effect of LAG3 and PD-1 on T cells in breast cancer patients are still not very clear. Here, with strict exclusion criteria, 88 patients with breast cancer and 18 healthy controls were enrolled. The percentages of LAG3⁺PD-1⁺ T cells in their peripheral blood (PBL) and tumor infiltrating T cells (TIL) were analyzed by flow cytometry, which showed an increase in TILs but no difference in PBLs and presented differences in TILs in different molecular subtypes (P < 0.05). In triple-negative breast cancer (TNBC), the highest percentages were observed, while in ER+/PR+ breast cancer, the lowest percentages were observed; however, these percentages were not different in different clinical stages (P > 0.05). Immunohistochemical staining showed that the expression of their ligands, PD-L1, MHC class II molecular and FGL1, was inconsistent in different molecular subtypes and clinical stages. Analysis of the functions of T cells with different phenotypes showed that the proliferation and secretion capacity of LAG3⁺PD-1⁺ T cells was obviously exhausted, with more than a two-fold of decrease compared with the groups of single positive LAG3 or PD-1 (P < 0.05). Finally, in a mouse model of TNBC, the dual blockade of LAG3 and PD-1 was indicated to achieve a better anti-tumour effect than either one alone (P < 0.05), which may provide a new strategy for the immunoregulatory treatment of patients with TNBC in the future.