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IntroductionPatients with frequent acute exacerbation phenotype chronic obstructive pulmonary disease (AECOPD) have a higher hospitalisation rate than infrequent exacerbation, the disease progresses quickly and treatment is more difficult. At present, it is impossible to predict patients with COPD with frequent acute exacerbation phenotypes. The composition of the lower respiratory tract flora and the intestinal flora is closely related to AECOPD, but the specific association mechanism between them is not very clear. This study used metagenomic next-generation sequencing (mNGS) technology to explore the microbial characteristics of the intestinal tract and airways of patients with COPD, and analyse the correlation between the sequencing results and inflammatory factors, immune factors and nutritional factors.Methods and analysisThis will be a prospective cohort study. We intend to recruit 152 patients with stable COPD. In the baseline, we will detect the participants’ induced sputum and faecal flora through mNGS, and changes in blood immune levels, and the patient’s condition is evaluated. Every 2 months, we will check the number of acute exacerbation through the phone range. After 12 months, we will check again the changes in the blood immune level, evaluate the patient’s condition and count the number of episodes.Ethics and disseminationThis study has been approved by the ethics committee of Guangdong Provincial Hospital of Traditional Chinese Medicine (approval number ZF2019-219-03). The results of the study will be published in peer-reviewed journals.Trial registration numberClinicalTrials.gov Registry (ChiCTR2000032870).

We present here INSIGHT (Isothermal NASBA-Sequencing based hIGH-througput Test): a two-stage COVID-19 testing strategy, using a combination of an isothermal NASBA reaction and next generation sequencing. From commercially acquired human saliva with spiked-in viral RNA as input, the first stage employs isothermal amplification of viral RNA to give a rapid result in one to two hours, using either fluorescence detection or a dipstick readout, whilst simultaneously incorporating sample-specific barcodes into the amplification product. In the first stage, fluorescent viral RNA detection can be consistently achieved at 10-100 copies per 20 µl reaction. The second stage pools post-amplification barcoded products from multiple samples for scalable sequencing that could be centralised, to further improve the accuracy of the test in a massively parallel way. Our two-stage testing strategy is suitable for further development into a home-based or point-of-care assay, and is potentially scalable to population level. IMPORTANT This protocol has not been validated on patient samples and should not be used for clinical diagnosis without further validation and certification. ***This is ongoing research to develop a testing strategy for the SARS-CoV-2 virus. The Wellcome Sanger Institute is not in a position to develop this into a commercial product, but would be open to discussions with third parties about how they could develop this further to achieve the societal benefits of this work.***

Background/Objectives: Bariatric surgery offers sustained marked weight loss and often remission of type 2 diabetes, yet the mechanisms of establishment of these health benefits are not clear. Subjects/Methods: We mapped the coordinated systemic responses of gut hormones, the circulating miRNAome and the metabolome in a rat model of Roux-en-Y gastric bypass (RYGB) surgery. Results: The response of circulating microRNAs (miRNAs) to RYGB was striking and selective. Analysis of 14 significantly altered circulating miRNAs within a pathway context was suggestive of modulation of signaling pathways including G protein signaling, neurodegeneration, inflammation, and growth and apoptosis responses. Concomitant alterations in the metabolome indicated increased glucose transport, accelerated glycolysis and inhibited gluconeogenesis in the liver. Of particular significance, we show significantly decreased circulating miRNA-122 levels and a more modest decline in hepatic levels, following surgery. In mechanistic studies, manipulation of miRNA-122 levels in a cell model induced changes in the activity of key enzymes involved in hepatic energy metabolism, glucose transport, glycolysis, tricarboxylic acid cycle, pentose phosphate shunt, fatty-acid oxidation and gluconeogenesis, consistent with the findings of the in vivo surgery-mediated responses, indicating the powerful homeostatic activity of the miRNAs. Conclusions: The close association between energy metabolism, neuronal signaling and gut microbial metabolites derived from the circulating miRNA, plasma, urine and liver metabolite and gut hormone correlations further supports an enhanced gut-brain signaling, which we suggest is hormonally mediated by both traditional gut hormones and miRNAs. This transomic approach to map the crosstalk between the circulating miRNAome and metabolome offers opportunities to understand complex systems biology within a disease and interventional treatment setting.

Bariatric surgery, also known as metabolic surgery, is an effective treatment for morbid obesity which also offers pronounced metabolic effects including the resolution of type 2 diabetes and a decrease in cardiovascular disease and long-term cancer risk. However, the mechanisms of surgical weight loss and the long term consequences of bariatric surgery remain unclear. Bariatric surgery has been demonstrated to alter the composition of both the microbiome and the metabolic phenotype. We observed a marked shift towards Gammaproteobacteria, particularly Enterobacter hormaechei, following Roux-en-Y gastric bypass (RYGB) surgery in a rat model compared with sham operated controls. Fecal water from RYGB surgery rats was highly cytotoxic to rodent cells (mouse lymphoma cell line), although In contrast, fecal water from sham operated animals showed no/very low cytotoxicity. This shift in the gross structure of the microbiome correlated with greatly increased cytotoxicity in a regulatory acceptable mouse lymphoma assay. Urinary phenylacetylglycine and indoxyl sulfate and fecal GABA, putrescine, tyramine and uracil were found to be inversely correlated with cell survival rate. This profound co-dependent response of mammalian and microbial metabolism to RYGB surgery and the impact on the cytotoxicity of the gut luminal environment suggests that RYGB exerts local and global metabolic effects which may have an influence on long term cancer risk and cytotoxic load.

Obesity is a worldwide public health issue which can lead to cardiovascular disease, type 2 diabetes, and cancers. Bariatric surgery can provide sustained weight loss and a significant reduction of cancer risk. MicroRNAs are approximately 22 nucleotide long post transcription regulators. MicroRNAs are suggested to play important roles in obesity and metabolic syndrome related biological processes, but also are involved in different types of cancer. Recently microRNAs have been shown to be present in blood, and it is proposed that circulating microRNAs may be biomarker of tissue biochemistry and pathology. The aim of this study is to investigate if the circulation microRNA profile is changed by bariatric surgery and whether the altered microRNA expression could contribute to the long term reduced predisposition cancer risk associated with such surgery. Total plasma RNA from 53 days post bariatric surgery and sham operated rats were isolated and then examined using Taqman Rodent MicroRNA A+B Cards which have the capacity to evaluate expression levels of 373 rat microRNAs. Four different normalization methods (Global, cel-miR-39 spiked in, U6 endogenous and five endogenous microRNAs normalization) were tested. Five endogenous microRNAs normalization was found to be optimal and selected for all downstream analysis. Our preliminary data showed that 113 microRNAs were expressed in the plasma of bariatric surgical rats and 92 in sham operated rats. Furthermore, an OPLS-DA model clearly showed that bariatric surgery changed the circulating microRNA expression profile. Among nineteen of the most significantly changed microRNAs, nine of them are relevant to cancer. In summary, our data suggest that bariatric surgery fundamentally changed circulating microRNA expression. Understanding the significance of these changes in circulating microRNA may give clues to the mechanisms whereby bariatric surgery affords health benefits to individual undergoing such surgery, including a reduced predisposition to disease such as cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4130. doi:1538-7445.AM2012-4130