Genetic Drivers of Heterogeneity in Type 2 Diabetes Pathophysiology

Type 2 diabetes (T2D) is a heterogeneous disease that develops through diverse pathophysiological processes1,2 and molecular mechanisms that are often specific to cell type3,4. Here, to characterize the genetic contribution to these processes across ancestry groups, we aggregate genome-wide association study data from 2,535,601 individuals (39.7% not of European ancestry), including 428,452 cases of T2D. We identify 1,289 independent association signals at genome-wide significance (P \u3c 5 × 10-8) that map to 611 loci, of which 145 loci are, to our knowledge, previously unreported. We define eight non-overlapping clusters of T2D signals that are characterized by distinct profiles of cardiometabolic trait associations. These clusters are differentially enriched for cell-type-specific regions of open chromatin, including pancreatic islets, adipocytes, endothelial cells and enteroendocrine cells. We build cluster-specific partitioned polygenic scores5 in a further 279,552 individuals of diverse ancestry, including 30,288 cases of T2D, and test their association with T2D-related vascular outcomes. Cluster-specific partitioned polygenic scores are associated with coronary artery disease, peripheral artery disease and end-stage diabetic nephropathy across ancestry groups, highlighting the importance of obesity-related processes in the development of vascular outcomes. Our findings show the value of integrating multi-ancestry genome-wide association study data with single-cell epigenomics to disentangle the aetiological heterogeneity that drives the development and progression of T2D. This might offer a route to optimize global access to genetically informed diabetes care

Genetic drivers of heterogeneity in type 2 diabetes pathophysiology

Type 2 diabetes (T2D) is a heterogeneous disease that develops through diverse pathophysiological processes1,2 and molecular mechanisms that are often specific to cell type3,4. Here, to characterize the genetic contribution to these processes across ancestry groups, we aggregate genome-wide association study data from 2,535,601 individuals (39.7% not of European ancestry), including 428,452 cases of T2D. We identify 1,289 independent association signals at genome-wide significance (P &lt; 5 × 10-8) that map to 611 loci, of which 145 loci are, to our knowledge, previously unreported. We define eight non-overlapping clusters of T2D signals that are characterized by distinct profiles of cardiometabolic trait associations. These clusters are differentially enriched for cell-type-specific regions of open chromatin, including pancreatic islets, adipocytes, endothelial cells and enteroendocrine cells. We build cluster-specific partitioned polygenic scores5 in a further 279,552 individuals of diverse ancestry, including 30,288 cases of T2D, and test their association with T2D-related vascular outcomes. Cluster-specific partitioned polygenic scores are associated with coronary artery disease, peripheral artery disease and end-stage diabetic nephropathy across ancestry groups, highlighting the importance of obesity-related processes in the development of vascular outcomes. Our findings show the value of integrating multi-ancestry genome-wide association study data with single-cell epigenomics to disentangle the aetiological heterogeneity that drives the development and progression of T2D. This might offer a route to optimize global access to genetically informed diabetes care.</p

The Effects of Oral Delivery of Recombinant House Dust Mite Allergen on Airway Inflammation in Murine Model of Asthma

氣喘是兒童常見的過敏性疾病之一，根據統計全球約一成的人口有氣喘的困擾，歐洲塵蟎 (Dermatophagoides pteronyssinus) 是八成台灣過敏病患的過敏原之一。氣喘的臨床症狀包含：呼吸道過度反應、嗜酸性白血球浸潤於呼吸道、活化的過敏原特異性第二型輔助型T細胞增加、呼吸道黏液增加、甚至是呼吸道重塑等等。口服耐受性，是指透過口服抗原而引發抗原特異性的免疫反應下降的現象，目前被認為具潛力成為過敏性氣喘的治療方式。本篇研究希望透過口服餵食基因重組第二類與第一類歐洲塵蟎過敏原 (recombinant D. pteronyssinus group 2 allergen, rDp2; recombinant D. pteronyssinus group 1 allergen, rDp1)，藉此來降低由歐洲塵蟎粗萃蛋白所引發的呼吸道發炎現象。我們利用腹腔注射粗萃蛋白致敏BALB/c雌性小鼠，並給予氣管粗萃蛋白引發呼吸道發炎。結果顯示，腹腔注射高劑量 (50 μg) 的粗萃蛋白可有效誘發過敏性氣喘的臨床症狀，包含血清中塵蟎蛋白特異性的IgE含量增加、促進嗜酸性白血球浸潤於肺部、肺部沖洗液中介白素-5 (Interleukin-5, IL-5) 含量增加、小鼠脾臟細胞經粗萃蛋白刺激可產生大量Th2的細胞激素 (IL-5與IL-13) 並且大量增生。更進一步，我們利用口服餵食的方式，連續七天給予致敏小鼠0.2或1.0 mg劑量的rDp2或粗萃蛋白，探討是否能有效降低粗萃蛋白引發的氣喘症狀。結果發現，給予1.0 mg的rDp2可有效降低呼吸道過度反應，並減緩呼吸道發炎現象。因此，我們認為餵食較高劑量的基因重組蛋白或粗萃蛋白，有減緩過敏性氣喘症狀的傾向；未來將需要更多的實驗，以尋找合適的餵食計量與時間。Asthma is one of the most common allergic diseases in children; in addition, about 80% of asthmatic patients in Taiwan are sensitized by house dust mite -- D. pteronyssinus. The characteristics of asthma such as AHR, eosinophils infiltration, antigen-specific T helper 2 cells activation, increased mucus secretion and even airway remodeling. Antigens-specific immune tolerance by prior oral administration of antigens might be a therapeutic strategy for allergic asthma. Therefore, we aimed to apply oral administration of rDp2 to decrease the airway inflammation induced by D. pteronyssinus. The female BALB/c mice were used and given with crude mite extract of D. pteronyssinus as the allergic asthma. In present study, we sensitized mice with peritoneal injection, and then challenged with intratracheal injection of crude mite extract. The results showed that peritoneal injection with high-dose could induce the clinical features of asthma significantly, including elevated mite-specific IgE in serum, production of Th2 cytokines (IL-5, IL-13) of splenocytes, and crude mite-specific lymphoproliferation. Furthermore, oral delivery of rDp2 or crude mite extract 0.2 or 1.0 mg/day for consecutive 7 days at the beginning of sensitization showed some beneficial effects on airway inflammation. Oral feeding 1.0 mg/day of rDp2 reduced AHR and slightly decreased the airway inflammation induced by crude mite extract. In conclusion, we suggest that oral delivery of high dose of single recombinant allergen seems to be more benefit on airway inflammation induced by the complex crude mite extract, and the feeding dose and feeding period need further investigation.Brief Contents文摘要 …………………………………………………………………… Ibstract …………………………………………………………………… IIrief Contents …………………………………………………………… IVull Contents ………………………………………………………… Vontents of Figures ………………………………………………………VIIIbbreviation ……………………………………………………………… IXntroduction ………………………………………………………… 1ypothesis & Specific Aims ………………………………………… 11aterials and Methods ……………………………………………… 13esults ………………………………………………………………… 28iscussion & Conclusion …………………………………………… 40igures ……………………………………………………………… 49eferences ……………………………………………………………70ppendix …………………………………………………………… 80ull Contents ntroduction ………………………………………………………………………… 1pidemiology of allergic asthma ……………………………………………… 2isk factors for asthma ………………………………………………………… 2efinition of asthma …………………………………………………………… 4echanism of asthma …………………………………………………………… 4nimal model for allergic asthma ……………………………………………… 6rugs for asthma treatment ……………………………………………………… 6ral tolerance …………………………………………………………………… 7ouse dust mite ………………………………………………………………… 9ecombinant allergens ………………………………………………………… 11ypothesis & Specific Aims ………………………………………………………… 11aterials and Methods ……………………………………………………………… 13reparation of crude mite extract ……………………………………………… 14reparation of rDp2 …………………………………………………………… 14xpression of rDp1 …………………………………………………………… 15estern blotting of rDp2 and rDp1 …………………………………………… 18nimals ………………………………………………………………………… 19ensitization and challenge of crude mite extract to establish allergic airway inflammation …………………………………………………………………… 19etermination of levels of crude mite specific antibodies in serum …………… 20etermination of levels of total IgE in serum ………………………………… 21easurement of AHR ………………………………………………………… 23nalysis of cellular composition of BALF …………………………………… 23etermination of cytokines secretion of splenocytes by ELISA ……………… 24etermination the lymphoproliferation of splenocytes ………………………… 26ral feeding protocol ………………………………………………………… 27tatistical analysis ……………………………………………………………… 27esults ……………………………………………………………………………… 28uccessful expression of rDp2 from a fermentor produced by Pichia pastoris .. 29xpression of rDp1 protein produced by Pichia pastoris ………………… 29ensitization with crude mite extract elevated specific immunoglobulin ……… 31ensitization and challenge with crude mite extract increased lymphoproliferation of splenocytes ………………………………………………………………… 32ensitization and challenge with crude mite extract increased inflammatory cytokine secretions of splenocytes …………………………………………… 33ensitization and challenge with crude mite extract induced AHR …………… 33ensitization and challenge with crude mite extract facilitated eosinophils infiltration in BALF …………………………………………………………… 34ral feeding showed no effects on the levels of immunoglobulin in serum …… 35he lymphoproliferation were no affected by oral feeding with mite allergens .. 36L-4 productions of splenocytes decreased after oral feeding ………………… 36ral feeding with mite allergens slightly elevated IL-10 levels of splenocytes .. 37ral feeding with higher dose of mite allergens reduced the AHR …………… 38he airway inflammation was slightly decreased after oral feeding …………… 38iscussion & Conclusion ………………………………………………………… 40llergic asthma model induction ……………………………………………… 41he effects of oral delivery with rDp2 on airway inflammation ……………… 43he effects of oral delivery with crude mite extract …………………………… 46onclusion ……………………………………………………………………… 47igures ……………………………………………………………………………… 49eferences ………………………………………………………………………… 60ppendix ………………………………………………………………………… 80ppendix 1. Schematic representation of vector maps of rDp1 or rDp2 ……… 81ppendix 2. Sequence of pPICZα-A-der p 1 ………………………………… 82ppendix 3. Reagents ………………………………………………………… 8

Van der Waals Epitaxial Growth of ZnO Films on Mica Substrates in Low-Temperature Aqueous Solution

In this article, we demonstrate the van der Waals (vdW) epitaxial growth of ZnO layers on mica substrates through a low-temperature hydrothermal process. The thermal pretreatment of mica substrates prior to the hydrothermal growth of ZnO is essential for growing ZnO crystals in epitaxy with the mica substrates. The addition of sodium citrate into the growth solution significantly promotes the growth of ZnO crystallites in a lateral direction to achieve fully coalesced, continuous ZnO epitaxial layers. As confirmed through transmission electron microscopy, the epitaxial paradigm of the ZnO layer on the mica substrate was regarded as an incommensurate van der Waals epitaxy. Furthermore, through the association of the Mist-CVD process, the high-density and uniform distribution of ZnO seeds preferentially occurred on mica substrates, leading to greatly improving the epitaxial qualities of the hydrothermally grown ZnO layers and obtaining flat surface morphologies. The electrical and optoelectrical properties of the vdW epitaxial ZnO layer grown on mica substrates were comparable with those grown on sapphire substrates through conventional solution-based epitaxy techniques

Anti-adhesive effect of hyaluronate in a rabbit laminectomy model

Background: Postlaminectomy dural adhesion is a common cause of recurrent symptoms. Hyaluronic acid-based gel has been reported to reduce the incidence of postoperative adhesion in the peritoneal cavity; however, its effect on preventing postoperative scar formation at laminectomy sites is not yet known. The purpose of this study was to evaluate the anti-adhesive effect of hyaluronic acid-based gelatin after laminectomy, using a rabbit model. Methods: Twelve adult New Zealand rabbits underwent two-level lumbar laminectomy, and were randomly assigned to one of two groups of six rabbits each. The treatment group received hyaluronic acid-based gelatin treatment and the control group was untreated. Rabbits were sacrificed 8 weeks after treatment. Peel-off testing and histological analysis were performed to assess the tenacity and the extent of adhesion formation. Results: No significant difference was observed in the neurologic performance between the two groups. The tenacity in the treatment group was significantly reduced compared to that of the control group (3.17 ± 0.75 vs. 4.33 ± 0.52, respectively; p = 0.016). Histological analysis showed significantly less scar tissue formation in the treatment group, with a larger subarachnoid space and greater distance between the dura and scar tissues. The amount of fibroblast cells also was significantly smaller in the treatment group than in the control group (3078 ± 313.68 vs. 3742 ± 455.65, respectively; p = 0.042). Conclusions: No serious adverse events were reported, and no difference was found in the incidence of complications between the treatment and control groups. The findings suggested that hyaluronic acid-based gelatin may be effective for preventing postlaminectomy dural adhesion in rabbits

Building a portable data and information interoperability infrastructure—framework for a standard Taiwan Electronic Medical Record Template

[[abstract]]Traditional electronic health record (EHR) data are produced from various hospital information systems. They could not have existed independently without an information system until the incarnation of XML technology. The interoperability of a healthcare system can be divided into two dimensions: functional interoperability and semantic interoperability. Currently, no single EHR standard exists that provides complete EHR interoperability. In order to establish a national EHR standard, we developed a set of local EHR templates. The Taiwan Electronic Medical Record Template (TMT) is a standard that aims to achieve semantic interoperability in EHR exchanges nationally. The TMT architecture is basically composed of forms, components, sections, and elements. Data stored in the elements which can be referenced by the code set, data type, and narrative block. The TMT was established with the following requirements in mind: (1) transformable to international standards; (2) having a minimal impact on the existing healthcare system; (3) easy to implement and deploy, and (4) compliant with Taiwan's current laws and regulations. The TMT provides a basis for building a portable, interoperable information infrastructure for EHR exchange in Taiwan

Segmental ureterectomy outcome of upper tract urothelial carcinoma in a high endemic area: A Taiwan nationwide collaborative study

Purpose:. According to the National Comprehensive Cancer Network guidelines, segmental ureterectomy (SU) of upper tract urothelial carcinoma (UTUC) is a considerable option for selected mid- and distal ureteral urothelial carcinoma (UC). As a UTUC endemic area, Taiwan lacks treatment outcome analysis of SU. Materials and methods:. This study retrospectively reviewed the treatment outcomes of SU for clinically localized UTUCs. Patients with biopsy or washing cytology-confirmed UTUCs who underwent open, laparoscopic, or robot-assisted management with curative intent were retrospectively reviewed for the eligibility of analysis. Cox regression was applied for univariable and multivariable analyses. Results:. A total of 161 patients who underwent SU were reviewed and analyzed. The median follow-up period was 44.5 (interquartile range, 21.6–84.9) months. After SU, 56/161 (34.8%) patients were free of UTUCs after the follow-up, 25/161 (15.5%) patients had local recurrence, and 35/161 (21.7%) had lymph node or distant metastasis. Surgical margin involvement was a risk factor associated with worse cancer-specific survival. Higher bladder recurrence and local recurrence rates were observed with concurrent bladder UC. Lymphovascular invasion and previous radical nephroureterectomy (RNU) for UC were related to higher local recurrence rates. Patients with pathological T3/T4 stage and end-stage renal disease tended to have higher metastasis rates. For the management of local recurrence, 19 patients received salvage RNU and 25 patients had adjuvant chemotherapy. However, 26/161 (16.1%) patients died of UTUCs and 2/161 (1.2%) patients died of surgery-related complications. Conclusion:. SU provides acceptable oncological outcomes if the surgeons select candidates carefully. SU is not recommended if the patient has T3 or higher stage or comorbidity of end-stage renal disease. Concurrent bladder UC is a risk factor for worse bladder recurrence-free survival and local recurrence-free survival. Lymphovascular invasion and previous RNU for UC were related to higher local recurrence rates. After SU, periodic follow-up is mandatory because the local recurrence rate is higher than radical surgery