Affiliated Eye Hospital of Wenzhou Medical College

AUTORES: Daniel J Klionsky1745,1749*, Kotb Abdelmohsen840, Akihisa Abe1237, Md Joynal Abedin1762, Hagai Abeliovich425,
\nAbraham Acevedo Arozena789, Hiroaki Adachi1800, Christopher M Adams1669, Peter D Adams57, Khosrow Adeli1981,
\nPeter J Adhihetty1625, Sharon G Adler700, Galila Agam67, Rajesh Agarwal1587, Manish K Aghi1537, Maria Agnello1826,
\nPatrizia Agostinis664, Patricia V Aguilar1960, Julio Aguirre-Ghiso784,786, Edoardo M Airoldi89,422, Slimane Ait-Si-Ali1376,
\nTakahiko Akematsu2010, Emmanuel T Akporiaye1097, Mohamed Al-Rubeai1394, Guillermo M Albaiceta1294,
\nChris Albanese363, Diego Albani561, Matthew L Albert517, Jesus Aldudo128, Hana Alg€ul1164, Mehrdad Alirezaei1198,
\nIraide Alloza642,888, Alexandru Almasan206, Maylin Almonte-Beceril524, Emad S Alnemri1212, Covadonga Alonso544,
\nNihal Altan-Bonnet848, Dario C Altieri1205, Silvia Alvarez1497, Lydia Alvarez-Erviti1395, Sandro Alves107,
\nGiuseppina Amadoro860, Atsuo Amano930, Consuelo Amantini1554, Santiago Ambrosio1458, Ivano Amelio756,
\nAmal O Amer918, Mohamed Amessou2089, Angelika Amon726, Zhenyi An1538, Frank A Anania291, Stig U Andersen6,
\nUsha P Andley2079, Catherine K Andreadi1690, Nathalie Andrieu-Abadie502, Alberto Anel2027, David K Ann58,
\nShailendra Anoopkumar-Dukie388, Manuela Antonioli832,858, Hiroshi Aoki1791, Nadezda Apostolova2007,
\nSaveria Aquila1500, Katia Aquilano1876, Koichi Araki292, Eli Arama2098, Agustin Aranda456, Jun Araya591,
\nAlexandre Arcaro1472, Esperanza Arias26, Hirokazu Arimoto1225, Aileen R Ariosa1749, Jane L Armstrong1930,
\nThierry Arnould1773, Ivica Arsov2120, Katsuhiko Asanuma675, Valerie Askanas1924, Eric Asselin1867, Ryuichiro Atarashi794,
\nSally S Atherton369, Julie D Atkin713, Laura D Attardi1131, Patrick Auberger1787, Georg Auburger379, Laure Aurelian1727,
\nRiccardo Autelli1992, Laura Avagliano1029,1755, Maria Laura Avantaggiati364, Limor Avrahami1166, Suresh Awale1986,
\nNeelam Azad404, Tiziana Bachetti568, Jonathan M Backer28, Dong-Hun Bae1933, Jae-sung Bae677, Ok-Nam Bae409,
\nSoo Han Bae2117, Eric H Baehrecke1729, Seung-Hoon Baek17, Stephen Baghdiguian1368,
\nAgnieszka Bagniewska-Zadworna2, Hua Bai90, Jie Bai667, Xue-Yuan Bai1133, Yannick Bailly884,
\nKithiganahalli Narayanaswamy Balaji473, Walter Balduini2002, Andrea Ballabio316, Rena Balzan1711, Rajkumar Banerjee239,
\nG abor B anhegyi1052, Haijun Bao2109, Benoit Barbeau1363, Maria D Barrachina2007, Esther Barreiro467, Bonnie Bartel997,
\nAlberto Bartolom e222, Diane C Bassham550, Maria Teresa Bassi1046, Robert C Bast Jr1273, Alakananda Basu1798,
\nMaria Teresa Batista1578, Henri Batoko1336, Maurizio Battino970, Kyle Bauckman2085, Bradley L Baumgarner1909,
\nK Ulrich Bayer1594, Rupert Beale1553, Jean-Fran¸cois Beaulieu1360, George R. Beck Jr48,294, Christoph Becker336,
\nJ David Beckham1595, Pierre-Andr e B edard749, Patrick J Bednarski301, Thomas J Begley1135, Christian Behl1419,
\nChristian Behrends757, Georg MN Behrens406, Kevin E Behrns1627, Eloy Bejarano26, Amine Belaid490,
\nFrancesca Belleudi1041, Giovanni B enard497, Guy Berchem706, Daniele Bergamaschi983, Matteo Bergami1401,
\nBen Berkhout1441, Laura Berliocchi714, Am elie Bernard1749, Monique Bernard1354, Francesca Bernassola1880,
\nAnne Bertolotti791, Amanda S Bess272, S ebastien Besteiro1351, Saverio Bettuzzi1828, Savita Bhalla913,
\nShalmoli Bhattacharyya973, Sujit K Bhutia838, Caroline Biagosch1159, Michele Wolfe Bianchi520,1378,1381,
\nMartine Biard-Piechaczyk210, Viktor Billes298, Claudia Bincoletto1314, Baris Bingol350, Sara W Bird1128, Marc Bitoun1112,
\nIvana Bjedov1258, Craig Blackstone843, Lionel Blanc1183, Guillermo A Blanco1496, Heidi Kiil Blomhoff1812,
\nEmilio Boada-Romero1297, Stefan B€ockler1464, Marianne Boes1423, Kathleen Boesze-Battaglia1835, Lawrence H Boise286,287,
\nAlessandra Bolino2063, Andrea Boman693, Paolo Bonaldo1823, Matteo Bordi897, J€urgen Bosch608, Luis M Botana1308,
\nJoelle Botti1375, German Bou1405, Marina Bouch e1038, Marion Bouchecareilh1331, Marie-Jos ee Boucher1901,
\nMichael E Boulton481, Sebastien G Bouret1926, Patricia Boya133, Micha€el Boyer-Guittaut1345, Peter V Bozhkov1141,
\nNathan Brady374, Vania MM Braga469, Claudio Brancolini1997, Gerhard H Braus353, Jos e M Bravo-San Pedro299,393,508,1374,
\nLisa A Brennan322, Emery H Bresnick2022, Patrick Brest490, Dave Bridges1939, Marie-Agn es Bringer124, Marisa Brini1822,
\nGlauber C Brito1311, Bertha Brodin631, Paul S Brookes1872, Eric J Brown352, Karen Brown1690, Hal E Broxmeyer480,
\nAlain Bruhat486,1339, Patricia Chakur Brum1893, John H Brumell446, Nicola Brunetti-Pierri315,1171,
\nRobert J Bryson-Richardson781, Shilpa Buch1777, Alastair M Buchan1819, Hikmet Budak1022, Dmitry V Bulavin118,505,1789,
\nScott J Bultman1792, Geert Bultynck665, Vladimir Bumbasirevic1470, Yan Burelle1356, Robert E Burke216,217,
\nMargit Burmeister1750, Peter B€utikofer1473, Laura Caberlotto1987, Ken Cadwell896, Monika Cahova112, Dongsheng Cai24,
\nJingjing Cai2099, Qian Cai1018, Sara Calatayud2007, Nadine Camougrand1343, Michelangelo Campanella1700,
\nGrant R Campbell1525, Matthew Campbell1249, Silvia Campello556,1876, Robin Candau1769, Isabella Caniggia1983,
\nLavinia Cantoni560, Lizhi Cao116, Allan B Caplan1656, Michele Caraglia1051, Claudio Cardinali1043, Sandra Morais Cardoso1579, Jennifer S Carew208, Laura A Carleton874, Cathleen R Carlin101, Silvia Carloni2002,
\nSven R Carlsson1267, Didac Carmona-Gutierrez1643, Leticia AM Carneiro312, Oliana Carnevali971, Serena Carra1318,
\nAlice Carrier120, Bernadette Carroll900, Caty Casas1324, Josefina Casas1116, Giuliana Cassinelli324, Perrine Castets1462,
\nSusana Castro-Obregon214, Gabriella Cavallini1841, Isabella Ceccherini568, Francesco Cecconi253,555,1884,
\nArthur I Cederbaum459, Valent ın Ce~na199,1281, Simone Cenci1323,2064, Claudia Cerella444, Davide Cervia1996,
\nSilvia Cetrullo1478, Hassan Chaachouay2028, Han-Jung Chae187, Andrei S Chagin634, Chee-Yin Chai626,628,
\nGopal Chakrabarti1502, Georgios Chamilos1601, Edmond YW Chan1142, Matthew TV Chan181, Dhyan Chandra1003,
\nPallavi Chandra548, Chih-Peng Chang818, Raymond Chuen-Chung Chang1653, Ta Yuan Chang345, John C Chatham1434,
\nSaurabh Chatterjee1910, Santosh Chauhan527, Yongsheng Che62, Michael E Cheetham1263, Rajkumar Cheluvappa1783,
\nChun-Jung Chen1153, Gang Chen598,1676, Guang-Chao Chen9, Guoqiang Chen1078, Hongzhuan Chen1077, Jeff W Chen1514,
\nJian-Kang Chen370,371, Min Chen249, Mingzhou Chen2104, Peiwen Chen1823, Qi Chen1674, Quan Chen172,
\nShang-Der Chen138, Si Chen325, Steve S-L Chen10, Wei Chen2125, Wei-Jung Chen829, Wen Qiang Chen979, Wenli Chen1113,
\nXiangmei Chen1133, Yau-Hung Chen1157, Ye-Guang Chen1250, Yin Chen1447, Yingyu Chen953,955, Yongshun Chen2135,
\nYu-Jen Chen712, Yue-Qin Chen1145, Yujie Chen1208, Zhen Chen339, Zhong Chen2123, Alan Cheng1702,
\nChristopher HK Cheng184, Hua Cheng1728, Heesun Cheong814, Sara Cherry1836, Jason Chesney1703,
\nChun Hei Antonio Cheung817, Eric Chevet1359, Hsiang Cheng Chi140, Sung-Gil Chi656, Fulvio Chiacchiera308,
\nHui-Ling Chiang958, Roberto Chiarelli1826, Mario Chiariello235,567,577, Marcello Chieppa835, Lih-Shen Chin290,
\nMario Chiong1285, Gigi NC Chiu878, Dong-Hyung Cho676, Ssang-Goo Cho650, William C Cho982, Yong-Yeon Cho105,
\nYoung-Seok Cho1064, Augustine MK Choi2095, Eui-Ju Choi656, Eun-Kyoung Choi387,400,685, Jayoung Choi1563,
\nMary E Choi2093, Seung-Il Choi2116, Tsui-Fen Chou412, Salem Chouaib395, Divaker Choubey1574, Vinay Choubey1936,
\nKuan-Chih Chow822, Kamal Chowdhury730, Charleen T Chu1856, Tsung-Hsien Chuang827, Taehoon Chun657,
\nHyewon Chung652, Taijoon Chung978, Yuen-Li Chung1194, Yong-Joon Chwae18, Valentina Cianfanelli254,
\nRoberto Ciarcia1775, Iwona A Ciechomska886, Maria Rosa Ciriolo1876, Mara Cirone1042, Sofie Claerhout1694,
\nMichael J Clague1698, Joan Cl aria1457, Peter GH Clarke1687, Robert Clarke361, Emilio Clementi1045,1398, C edric Cleyrat1781,
\nMiriam Cnop1366, Eliana M Coccia574, Tiziana Cocco1459, Patrice Codogno1375, J€orn Coers271, Ezra EW Cohen1533,
\nDavid Colecchia235,567,577, Luisa Coletto25, N uria S Coll123, Emma Colucci-Guyon516, Sergio Comincini1829,
\nMaria Condello578, Katherine L Cook2073, Graham H Coombs1929, Cynthia D Cooper2076, J Mark Cooper1395,
\nIsabelle Coppens601, Maria Tiziana Corasaniti1387, Marco Corazzari485,1884, Ramon Corbalan1566,
\nElisabeth Corcelle-Termeau251, Mario D Cordero1899, Cristina Corral-Ramos1289, Olga Corti507,1109, Andrea Cossarizza1767,
\nPaola Costelli1993, Safia Costes1518, Susan L Cotman721, Ana Coto-Montes946, Sandra Cottet566,1688, Eduardo Couve1301,
\nLori R Covey1015, L Ashley Cowart762, Jeffery S Cox1536, Fraser P Coxon1427, Carolyn B Coyne1846, Mark S Cragg1919,
\nRolf J Craven1679, Tiziana Crepaldi1995, Jose L Crespo1300, Alfredo Criollo1285, Valeria Crippa558, Maria Teresa Cruz1576,
\nAna Maria Cuervo26, Jose M Cuezva1277, Taixing Cui1907, Pedro R Cutillas987, Mark J Czaja27, Maria F Czyzyk-Krzeska1572,
\nRuben K Dagda2068, Uta Dahmen1404, Chunsun Dai800, Wenjie Dai1187, Yun Dai2059, Kevin N Dalby1940,
\nLuisa Dalla Valle1822, Guillaume Dalmasso1340, Marcello D’Amelio557, Markus Damme188, Arlette Darfeuille-Michaud1340,
\nCatherine Dargemont950, Victor M Darley-Usmar1433, Srinivasan Dasarathy205, Biplab Dasgupta202, Srikanta Dash1254,
\nCrispin R Dass242, Hazel Marie Davey8, Lester M Davids1560, David D avila227, Roger J Davis1731, Ted M Dawson604,
\nValina L Dawson606, Paula Daza1898, Jackie de Belleroche470, Paul de Figueiredo1180,1182,
\nRegina Celia Bressan Queiroz de Figueiredo135, Jos e de la Fuente1023, Luisa De Martino1775,
\nAntonella De Matteis1171, Guido RY De Meyer1443, Angelo De Milito631, Mauro De Santi2002,

autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.

Type 2 diabetes (T2D) is a very common disease in humans. Here we conduct a meta-analysis of genome-wide association studies (GWAS) with ~16 million genetic variants in 62,892 T2D cases and 596,424 controls of European ancestry. We identify 139 common and 4 rare variants associated with T2D, 42 of which (39 common and 3 rare variants) are independent of the known variants. Integration of the gene expression data from blood (n = 14,115 and 2765) with the GWAS results identifies 33 putative functional genes for T2D, 3 of which were targeted by approved drugs. A further integration of DNA methylation (n = 1980) and epigenomic annotation data highlight 3 genes (CAMK1D, TP53INP1, and ATP5G1) with plausible regulatory mechanisms, whereby a genetic variant exerts an effect on T2D through epigenetic regulation of gene expression. Our study uncovers additional loci, proposes putative genetic regulatory mechanisms for T2D, and provides evidence of purifying selection for T2D-associated variants.

Health risk factors such as body mass index (BMI) and serum cholesterol are associated with many common diseases. It often remains unclear whether the risk factors are cause or consequence of disease, or whether the associations are the result of confounding. We develop and apply a method (called GSMR) that performs a multi-SNP Mendelian randomization analysis using summary-level data from genome-wide association studies to test the causal associations of BMI, waist-to-hip ratio, serum cholesterols, blood pressures, height, and years of schooling (EduYears) with common diseases (sample sizes of up to 405,072). We identify a number of causal associations including a protective effect of LDL-cholesterol against type-2 diabetes (T2D) that might explain the side effects of statins on T2D, a protective effect of EduYears against Alzheimer's disease, and bidirectional associations with opposite effects (e.g., higher BMI increases the risk of T2D but the effect of T2D on BMI is negative).

Abstract Common single-nucleotide polymorphisms (SNPs) are predicted to collectively explain 40–50% of phenotypic variation in human height, but identifying the specific variants and associated regions requires huge sample sizes 1 . Here, using data from a genome-wide association study of 5.4 million individuals of diverse ancestries, we show that 12,111 independent SNPs that are significantly associated with height account for nearly all of the common SNP-based heritability. These SNPs are clustered within 7,209 non-overlapping genomic segments with a mean size of around 90 kb, covering about 21% of the genome. The density of independent associations varies across the genome and the regions of increased density are enriched for biologically relevant genes. In out-of-sample estimation and prediction, the 12,111 SNPs (or all SNPs in the HapMap 3 panel 2 ) account for 40% (45%) of phenotypic variance in populations of European ancestry but only around 10–20% (14–24%) in populations of other ancestries. Effect sizes, associated regions and gene prioritization are similar across ancestries, indicating that reduced prediction accuracy is likely to be explained by linkage disequilibrium and differences in allele frequency within associated regions. Finally, we show that the relevant biological pathways are detectable with smaller sample sizes than are needed to implicate causal genes and variants. Overall, this study provides a comprehensive map of specific genomic regions that contain the vast majority of common height-associated variants. Although this map is saturated for populations of European ancestry, further research is needed to achieve equivalent saturation in other ancestries.

The identification of genes and regulatory elements underlying the associations discovered by GWAS is essential to understanding the aetiology of complex traits (including diseases). Here, we demonstrate an analytical paradigm of prioritizing genes and regulatory elements at GWAS loci for follow-up functional studies. We perform an integrative analysis that uses summary-level SNP data from multi-omics studies to detect DNA methylation (DNAm) sites associated with gene expression and phenotype through shared genetic effects (i.e., pleiotropy). We identify pleiotropic associations between 7858 DNAm sites and 2733 genes. These DNAm sites are enriched in enhancers and promoters, and >40% of them are mapped to distal genes. Further pleiotropic association analyses, which link both the methylome and transcriptome to 12 complex traits, identify 149 DNAm sites and 66 genes, indicating a plausible mechanism whereby the effect of a genetic variant on phenotype is mediated by genetic regulation of transcription through DNAm.

A major challenge for effective application of CRISPR systems is to accurately predict the single guide RNA (sgRNA) on-target knockout efficacy and off-target profile, which would facilitate the optimized design of sgRNAs with high sensitivity and specificity. Here we present DeepCRISPR, a comprehensive computational platform to unify sgRNA on-target and off-target site prediction into one framework with deep learning, surpassing available state-of-the-art in silico tools. In addition, DeepCRISPR fully automates the identification of sequence and epigenetic features that may affect sgRNA knockout efficacy in a data-driven manner. DeepCRISPR is available at http://www.deepcrispr.net/ .

For the last 20 years, a great amount of evidence has accumulated through epidemiological studies that most of the dry eye disease encountered in daily life, especially in video display terminal (VDT) workers, involves short tear film breakup time (TFBUT) type dry eye, a category characterized by severe symptoms but minimal clinical signs other than short TFBUT. An unstable tear film also affects the visual function, possibly due to the increase of higher order aberrations. Based on the change in the understanding of the types, symptoms, and signs of dry eye disease, the Asia Dry Eye Society agreed to the following definition of dry eye: "Dry eye is a multifactorial disease characterized by unstable tear film causing a variety of symptoms and/or visual impairment, potentially accompanied by ocular surface damage." The definition stresses instability of the tear film as well as the importance of visual impairment, highlighting an essential role for TFBUT assessment. This paper discusses the concept of Tear Film Oriented Therapy (TFOT), which evolved from the definition of dry eye, emphasizing the importance of a stable tear film.

Biofilms that contribute to the persistent bacterial infections pose serious threats to global public health, mainly due to their resistance to antibiotics penetration and escaping innate immune attacks by phagocytes. Here, we report a kind of surface-adaptive gold nanoparticles (AuNPs) exhibiting (1) a self-adaptive target to the acidic microenvironment of biofilm, (2) an enhanced photothermal ablation of methicillin-resistant Staphylococcus aureus (MRSA) biofilm under near-infrared (NIR) light irradiation, and (3) no damage to the healthy tissues around the biofilm. Originally, AuNPs were readily prepared by surface modification with pH-responsive mixed charged zwitterionic self-assembled monolayers consisting of weak electrolytic 11-mercaptoundecanoic acid (HS-C10-COOH) and strong electrolytic (10-mercaptodecyl)trimethylammonium bromide (HS-C10-N4). The mixed charged zwitterion-modified AuNPs showed fast pH-responsive transition from negative charge to positive charge, which enabled the AuNPs to disperse well in healthy tissues (pH ∼7.4), while quickly presenting strong adherence to negatively charged bacteria surfaces in MRSA biofilm (pH ∼5.5). Simultaneous AuNP aggregation within the MRSA biofilm enhanced the photothermal ablation of MRSA biofilm under NIR light irradiation. The surrounding healthy tissues showed no damage because the dispersed AuNPs had no photothermal effect under NIR light. In view of the above advantages as well as the straightforward preparation, AuNPs developed in this work may find potential applications as a useful antibacterial agent in the areas of healthcare.

) promoted myofibroblast transdifferentiation with down-regulation of type I collagen in human scleral fibroblasts. Importantly, the antihypoxia drugs salidroside and formononetin down-regulated HIF-1α expression as well as the phosphorylation levels of eIF2α and mTOR, slowing experimental myopia progression without affecting normal ocular growth in guinea pigs. Furthermore, eIF2α phosphorylation inhibition suppressed experimental myopia, whereas mTOR phosphorylation induced myopia in normal mice. Collectively, these findings defined an essential role of hypoxia in scleral ECM remodeling and myopia development, suggesting a therapeutic approach to control myopia by ameliorating hypoxia.

Polydopamine (PDA) nanoparticles have emerged as an attractive biomimetic photothermal agent in photothermal antibacterial therapy due to their ease of synthesis, good biodegradability, long-term safety, and excellent photostability. However, the therapeutic effects of PDA nanoparticles are generally limited by the low photothermal conversion efficiency (PCE). Herein, PDA@Ag nanoparticles are synthesized via growing Ag on the surface of PDA nanoparticles and then encapsulated into a cationic guar gum (CG) hydrogel network. The optimized CG/PDA@Ag platform exhibits a high PCE (38.2%), which is more than two times higher than that of pure PDA (16.6%). More importantly, the formulated CG/PDA@Ag hydrogel with many active groups can capture and kill bacteria through effective interactions between hydrogel and bacteria, thereby benefiting the antibacterial effect. As anticipated, the designed CG/PDA@Ag system combined the advantages of PDA@Ag nanoparticles (high PCE) and hydrogel (preventing aggregation of PDA@Ag nanoparticles and possessing inherent antibacterial ability) is demonstrated to have superior antibacterial efficacy both in vitro and in vivo. This study develops a facile approach to boost the PCE of PDA for photothermal antibacterial therapy, providing a significant step forward in advancing the application of PDA nano-photothermal agents.

Abstract Understanding the difference in genetic regulation of gene expression between brain and blood is important for discovering genes for brain-related traits and disorders. Here, we estimate the correlation of genetic effects at the top-associated cis -expression or -DNA methylation (DNAm) quantitative trait loci ( cis -eQTLs or cis -mQTLs) between brain and blood ( r b ). Using publicly available data, we find that genetic effects at the top cis -eQTLs or mQTLs are highly correlated between independent brain and blood samples ( $$\hat r_b = 0.70$$ <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"> <mml:msub> <mml:mrow> <mml:mover> <mml:mrow> <mml:mi>r</mml:mi> </mml:mrow> <mml:mo>^</mml:mo> </mml:mover> </mml:mrow> <mml:mrow> <mml:mi>b</mml:mi> </mml:mrow> </mml:msub> <mml:mo>=</mml:mo> <mml:mn>0.70</mml:mn> </mml:math> for cis -eQTLs and $$\hat r_ b = 0.78$$ <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"> <mml:msub> <mml:mrow> <mml:mover> <mml:mrow> <mml:mi>r</mml:mi> </mml:mrow> <mml:mo>^</mml:mo> </mml:mover> </mml:mrow> <mml:mrow> <mml:mi>b</mml:mi> </mml:mrow> </mml:msub> <mml:mo>=</mml:mo> <mml:mn>0.78</mml:mn> </mml:math> for cis -mQTLs). Using meta-analyzed brain cis -eQTL/mQTL data ( n = 526 to 1194), we identify 61 genes and 167 DNAm sites associated with four brain-related phenotypes, most of which are a subset of the discoveries (97 genes and 295 DNAm sites) using data from blood with larger sample sizes ( n = 1980 to 14,115). Our results demonstrate the gain of power in gene discovery for brain-related phenotypes using blood cis -eQTL/mQTL data with large sample sizes.

Gelatin methacryloyl (GelMA) is a versatile material for a wide range of bioapplications. There is an intense interest in developing effective chemical strategies to prepare GelMA with a high degree of batch-to-batch consistency and controllability in terms of methacryloyl functionalization and physiochemical properties. Herein, we systematically investigated the batch-to-batch reproducibility and controllability of producing GelMA (target highly and lowly substituted versions) via a one-pot strategy. To assess the GelMA product, several parameters were evaluated, including the degree of methacryloylation, secondary structure, and enzymatic degradation, along with the mechanical properties and cell viability of GelMA hydrogels. The results showed that two types of target GelMA with five batches exhibited a high degree of controllability and reproducibility in compositional, structural, and functional properties owing to the highly controllable one-pot strategy.

In the face of an "epidemic" increase in myopia over the last decades and myopia prevalence predicted to reach 2.5 billion people by the end of this decade, there is an urgent need to develop effective and safe therapeutic interventions to slow down this "myopia booming" and prevent myopia-related complications and vision loss. Dopamine (DA) is an important neurotransmitter in the retina and mediates diverse functions including retina development, visual signaling, and refractive development. Inspired by the convergence of epidemiological and animal studies in support of the inverse relationship between outdoor activity and risk of developing myopia and by the close biological relationship between light exposure and dopamine release/signaling, we felt it is timely and important to critically review the role of DA in myopia development. This review will revisit several key points of evidence for and against DA mediating light control of myopia: 1) the causal role of extracellular retinal DA levels, 2) the mechanism and action of dopamine D1 and D2 receptors and 3) the roles of cellular/circuit retinal pathways. We examine the experiments that show causation by altering DA, DA receptors and visual pathways using pharmacological, transgenic, or visual environment approaches. Furthermore, we critically evaluate the safety issues of a DA-based treatment strategy and some approaches to address these issues. The review identifies the key questions and challenges in translating basic knowledge on DA signaling and myopia from animal studies into effective pharmacological treatments for myopia in children.

Current therapeutic protocols for diabetic foot ulcers (DFUs), a severe and rapidly growing chronic complication in diabetic patients, remain nonspecific. Hyperglycemia-caused inflammation and excessive reactive oxygen species (ROS) are common obstacles encountered in DFU wound healing, often leading to impaired recovery. These two effects reinforce each other, forming an endless loop. However, adequate and inclusive methods are still lacking to target these two aspects and break the vicious cycle. This study proposes a novel approach for treating DFU wounds, utilizing an immunomodulatory hydrogel to achieve self-cascade glucose depletion and ROS scavenging to regulate the diabetic microenvironment. Specifically, AuPt@melanin-incorporated (GHM3) hydrogel dressing is developed to facilitate efficient hyperthermia-enhanced local glucose depletion and ROS scavenging. Mechanistically, in vitro/vivo experiments and RNA sequencing analysis demonstrate that GHM3 disrupts the ROS-inflammation cascade cycle and downregulates the ratio of M1/M2 macrophages, consequently improving the therapeutic outcomes for dorsal skin and DFU wounds in diabetic rats. In conclusion, this proposed approach offers a facile, safe, and highly efficient treatment modality for DFUs.

BACKGROUND: Increasing evidence has demonstrated the functional relevance of long non-coding RNAs (lncRNAs) to immunity regulation and the tumor microenvironment in non-small cell lung cancer (NSCLC). However, tumor immune infiltration-associated lncRNAs and their value in improving clinical outcomes and immunotherapy remain largely unexplored. METHODS: We developed a computational approach to identify an lncRNA signature (TILSig) as an indicator of immune cell infiltration in patients with NSCLC through integrative analysis for lncRNA, immune and clinical profiles of 115 immune cell lines, 187 NSCLC cell lines and 1533 patients with NSCLC. Then the influence of the TILSig on the prognosis and immunotherapy in NSCLC was comprehensively investigated. RESULTS: Computational immune and lncRNA profiling analysis identified an lncRNA signature (TILSig) consisting of seven lncRNAs associated with tumor immune infiltration. The TILSig significantly stratified patients into the immune-cold group and immune-hot group in both training and validation cohorts. These immune-hot patients exhibit significantly improved survival outcome and greater immune cell infiltration compared with immune-cold patients. Multivariate analysis revealed that the TILSig is an independent predictive factor after adjusting for other clinical factors. Further analysis accounting for TILSig and immune checkpoint gene revealed that the TILSig has a discriminatory power in patients with similar expression levels of immune checkpoint genes and significantly prolonged survival was observed for patients with low TILSig and low immune checkpoint gene expression implying a better response to immune checkpoint inhibitor (ICI) immunotherapy. CONCLUSIONS: Our finding demonstrated the importance and value of lncRNAs in evaluating the immune infiltrate of the tumor and highlighted the potential of lncRNA coupled with specific immune checkpoint factors as predictive biomarkers of ICI response to enable a more precise selection of patients.

MicroRNAs (miRNAs) are endogenous short (∼22) nucleotide RNAs that regulate gene function by modification of target mRNAs. miRNA-1 (miR-1) and miRNA-206 (miR-206) are highly expressed in skeletal muscle. Due to the tissue-specific nature of miR-1/206 for skeletal muscles, we investigated the role of miR-1/206 in the development of rhabdomyosarcoma. Initially, we demonstrated that miR-1/206 expression was suppressed in rhabdomyosarcomas and found at very low levels in a rhabdomyosarcoma RD cell line. Transient transfection of miR-1/206 into cultured RD cells led to a significant decrease in cell growth and migration. Using bioinformatics, we identified two putative miR-1/206 binding sites within the 3′-untranslated region of the human c-Met mRNA. miR-1/206 was then shown to have activity on mRNA expression by targeting the c-Met 3′-untranslated region. The expression of c-Met protein was shown to be down-regulated by subsequent Western blot analysis. Conversely, up-regulation of c-Met was confirmed in tissue samples of human rhabdomyosarcoma, with its level inversely correlated with miR-1/206 expression. In vivo, miR-1/206-expressing tumor cells showed growth delay in comparison with negative control. Our results demonstrated that miR-1/206 suppressed c-Met expression in rhabdomyosarcoma and could function as a potent tumor suppressor in c-Met-overexpressing tumors. Inhibition of miR-1/206 function could contribute to aberrant cell proliferation and migration, leading to rhabdomyosarcoma development. MicroRNAs (miRNAs) are endogenous short (∼22) nucleotide RNAs that regulate gene function by modification of target mRNAs. miRNA-1 (miR-1) and miRNA-206 (miR-206) are highly expressed in skeletal muscle. Due to the tissue-specific nature of miR-1/206 for skeletal muscles, we investigated the role of miR-1/206 in the development of rhabdomyosarcoma. Initially, we demonstrated that miR-1/206 expression was suppressed in rhabdomyosarcomas and found at very low levels in a rhabdomyosarcoma RD cell line. Transient transfection of miR-1/206 into cultured RD cells led to a significant decrease in cell growth and migration. Using bioinformatics, we identified two putative miR-1/206 binding sites within the 3′-untranslated region of the human c-Met mRNA. miR-1/206 was then shown to have activity on mRNA expression by targeting the c-Met 3′-untranslated region. The expression of c-Met protein was shown to be down-regulated by subsequent Western blot analysis. Conversely, up-regulation of c-Met was confirmed in tissue samples of human rhabdomyosarcoma, with its level inversely correlated with miR-1/206 expression. In vivo, miR-1/206-expressing tumor cells showed growth delay in comparison with negative control. Our results demonstrated that miR-1/206 suppressed c-Met expression in rhabdomyosarcoma and could function as a potent tumor suppressor in c-Met-overexpressing tumors. Inhibition of miR-1/206 function could contribute to aberrant cell proliferation and migration, leading to rhabdomyosarcoma development. Soft tissue sarcomas are a heterogeneous group of mesenchymal tumors that carries a guarded prognosis due to the aggressive local invasion and metastatic potential of these tumors. Progress in the search for etiology and treatment of soft tissue sarcomas is hampered by the fact that they represent a small proportion of all malignancies. Rhabdomyosarcoma (RMS) 3The abbreviations used are: RMSrhabdomyosarcomamiRNAmicroRNAHGFhepatocyte growth factorDIGdigoxigeninGAPDHglyceraldehyde-3-phosphate dehydrogenaseUTRuntranslated regionDMEMDulbecco's modified Eagle's mediumMTS3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner saltERK1/2extracellular signal-regulated kinases 1 and 2FAKfocal adhesion kinaseNCnegative control. is a distinct soft tissue sarcoma that likely originates from cells of the myogenic lineage (1Breitfeld P.P. Meyer W.H. Oncologist. 2005; 10: 518-527Crossref PubMed Scopus (97) Google Scholar). On the basis of histology, three main subgroups are often described: alveolar, embryonal, and pleomorphic RMS (2Ferrari A. Dileo P. Casanova M. Bertulli R. Meazza C. Gandola L. Navarria P. Collini P. Gronchi A. Olmi P. Fossati-Bellani F. Casali P.G. Cancer. 2003; 98: 571-580Crossref PubMed Scopus (293) Google Scholar). Alveolar RMS consists of small, round, and densely packed cells and occurs mainly in the trunk and extremities and carries with it a more unfavorable prognosis (1Breitfeld P.P. Meyer W.H. Oncologist. 2005; 10: 518-527Crossref PubMed Scopus (97) Google Scholar, 3Taulli R. Scuoppo C. Bersani F. Accornero P. Forni P.E. Miretti S. Grinza A. Allegra P. Schmitt-Ney M. Crepaldi T. Ponzetto C. Cancer Res. 2006; 66: 4742-4749Crossref PubMed Scopus (135) Google Scholar). Embryonal RMS typically consists of spindle-shaped cells and occurs mainly in the head and neck region (1Breitfeld P.P. Meyer W.H. Oncologist. 2005; 10: 518-527Crossref PubMed Scopus (97) Google Scholar, 3Taulli R. Scuoppo C. Bersani F. Accornero P. Forni P.E. Miretti S. Grinza A. Allegra P. Schmitt-Ney M. Crepaldi T. Ponzetto C. Cancer Res. 2006; 66: 4742-4749Crossref PubMed Scopus (135) Google Scholar). Pleomorphic RMS is uncommon and usually occurs in the adult population (2Ferrari A. Dileo P. Casanova M. Bertulli R. Meazza C. Gandola L. Navarria P. Collini P. Gronchi A. Olmi P. Fossati-Bellani F. Casali P.G. Cancer. 2003; 98: 571-580Crossref PubMed Scopus (293) Google Scholar). Until now, treatment of RMS has largely been based on local regional control and toxic systemic chemotherapy regimens without directed cellular therapy (2Ferrari A. Dileo P. Casanova M. Bertulli R. Meazza C. Gandola L. Navarria P. Collini P. Gronchi A. Olmi P. Fossati-Bellani F. Casali P.G. Cancer. 2003; 98: 571-580Crossref PubMed Scopus (293) Google Scholar, 3Taulli R. Scuoppo C. Bersani F. Accornero P. Forni P.E. Miretti S. Grinza A. Allegra P. Schmitt-Ney M. Crepaldi T. Ponzetto C. Cancer Res. 2006; 66: 4742-4749Crossref PubMed Scopus (135) Google Scholar). Recent investigations, however, are improving our understanding of its tumor biology and are helping to identify novel prognostic factors and targets for clinical therapy (1Breitfeld P.P. Meyer W.H. Oncologist. 2005; 10: 518-527Crossref PubMed Scopus (97) Google Scholar). rhabdomyosarcoma microRNA hepatocyte growth factor digoxigenin glyceraldehyde-3-phosphate dehydrogenase untranslated region Dulbecco's modified Eagle's medium 3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt extracellular signal-regulated kinases 1 and 2 focal adhesion kinase negative control. MicroRNAs (miRNAs) are endogenously expressed, non-protein-coding RNAs that can influence a wide variety of biological processes including development, metabolism, proliferation, differentiation, and oncogenesis (4He L. Hannon G.J. Nat. Rev. Genet. 2004; 5: 522-531Crossref PubMed Scopus (5701) Google Scholar). Aberrant post-transcriptional regulation of mRNAs by miRNAs can lead to oncogenesis with increased cell proliferation, decreased apoptosis, and enhanced metastatic potential of affected cells (5Zhang B. Pan X. Cobb G.P. Anderson T.A. Dev. Biol. 2007; 302: 1-12Crossref PubMed Scopus (2204) Google Scholar). Following the discovery of miRNA let-7, multiple miRNAs were linked to oncogenes and tumor suppressor genes including the Ras proto-oncogene, the antiapoptotic gene BCL2, the potent p53 tumor suppressor gene, and the MET oncogene (5Zhang B. Pan X. Cobb G.P. Anderson T.A. Dev. Biol. 2007; 302: 1-12Crossref PubMed Scopus (2204) Google Scholar, 6Cho W.C. Mol. Cancer. 2007; 6: 60Crossref PubMed Scopus (638) Google Scholar). The MET oncogene encodes a cell surface receptor tyrosine kinase, c-Met, that is up-regulated in a variety of tumors including rhabdomyosarcoma (7Birchmeier C. Birchmeier W. Gherardi E. Vande Woude G.F. Nat. Rev. Mol. Cell Biol. 2003; 4: 915-925Crossref PubMed Scopus (2240) Google Scholar, 8Mazzone M. Comoglio P.M. FASEB J. 2006; 20: 1611-1621Crossref PubMed Scopus (111) Google Scholar). c-Met is a disulfide-linked heterodimer consisting of an extracellular α-subunit and a β-subunit that spans the plasma membrane and contains the catalytic region with tyrosine kinase activity (7Birchmeier C. Birchmeier W. Gherardi E. Vande Woude G.F. Nat. Rev. Mol. Cell Biol. 2003; 4: 915-925Crossref PubMed Scopus (2240) Google Scholar). Binding of hepatocyte growth factor (HGF)/scatter factor induces c-Met dimerization and autophosphorylation, which leads to cellular activation (7Birchmeier C. Birchmeier W. Gherardi E. Vande Woude G.F. Nat. Rev. Mol. Cell Biol. 2003; 4: 915-925Crossref PubMed Scopus (2240) Google Scholar). c-Met activation, through aberrant HGF stimulation, can contribute to tumor growth, invasiveness, and metastasis (9Danilkovitch-Miagkova A. Zbar B. J. Clin. Invest. 2002; 109: 863-867Crossref PubMed Scopus (268) Google Scholar). c-Met has been predicted and shown to be the target gene of multiple miRNAs including miRNA-206 (miR-206) (10McCarthy J.J. Biochim. Biophys. Acta. 2008; 1779: 682-691Crossref PubMed Scopus (322) Google Scholar). miR-206 and miRNA-1 (miR-1) are members of the muscle-specific miR-1 family of so-called myomiRs that currently consists of six members (10McCarthy J.J. Biochim. Biophys. Acta. 2008; 1779: 682-691Crossref PubMed Scopus (322) Google Scholar). Based on sequence conservation of the seed region, the miR-1 family can be divided into miR-1/206 or miR-133a/b subgroups (11Chen J.F. Mandel E.M. Thomson J.M. Wu Q. Callis T.E. Hammond S.M. Conlon F.L. Wang D.Z. Nat. Genet. 2006; 38: 228-233Crossref PubMed Scopus (2254) Google Scholar, 12Kim H.K. Lee Y.S. Sivaprasad U. Malhotra A. Dutta A. J. Cell Biol. 2006; 174: 677-687Crossref PubMed Scopus (653) Google Scholar). Although miR-1 is highly enriched in both cardiac and skeletal muscle, miR-206 is exclusively expressed in skeletal muscle (11Chen J.F. Mandel E.M. Thomson J.M. Wu Q. Callis T.E. Hammond S.M. Conlon F.L. Wang D.Z. Nat. Genet. 2006; 38: 228-233Crossref PubMed Scopus (2254) Google Scholar, 12Kim H.K. Lee Y.S. Sivaprasad U. Malhotra A. Dutta A. J. Cell Biol. 2006; 174: 677-687Crossref PubMed Scopus (653) Google Scholar). Therefore, it can be deduced that these miRNAs play an important role in the myogenesis and development of cardiac and skeletal muscles. miR-1 knock-out mice confirmed that this miRNA is necessary for cardiac development and physiology (13Zhao Y. Ransom J.F. Li A. Vedantham V. von Drehle M. Muth A.N. Tsuchihashi T. McManus M.T. Schwartz R.J. Srivastava D. Cell. 2007; 129: 303-317Abstract Full Text Full Text PDF PubMed Scopus (1196) Google Scholar). miR-206, which is specifically expressed in skeletal muscle and rarely detected in the heart, plays an important role in skeletal muscle development (12Kim H.K. Lee Y.S. Sivaprasad U. Malhotra A. Dutta A. J. Cell Biol. 2006; 174: 677-687Crossref PubMed Scopus (653) Google Scholar, 14Anderson C. Catoe H. Werner R. Nucleic Acids Res. 2006; 34: 5863-5871Crossref PubMed Scopus (327) Google Scholar). However, development and progression of skeletal muscle tumors such as rhabdomyosarcoma based on the presence or absence of miR-1 and miR-206 remain largely unknown. In this study, we attempted to decipher the biological function of miR-1/206 in human rhabdomyosarcoma specimens and rhabdomyosarcoma RD cells. By causal association following the identification of cell specificity for miR-1/206, we surmised that miR-1/206 acted as a suppressor of RD cell proliferation and migration. Furthermore, we set out to elucidate the cellular mechanisms responsible for its activity and to identify its target, c-Met, so that it may one day serve as a potential target in the treatment of rhabdomyosarcoma. The human rhabdomyosarcoma cell line, RD, purchased from ATCC (Manassas, VA), was grown in Dulbecco's modified Eagle's medium (DMEM; Invitrogen) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT) and incubated at 37 °C in a humidified incubator containing 5% CO2. HEK-293 cells were grown under the same conditions. Eight rhabdomyosarcoma specimens and normal donor skeletal muscle tissues were obtained from the Eye Hospital and the First Affiliated Hospital of Wenzhou Medical College (Wenzhou, China). All studies and procedures involving human tissue samples were approved by the Wenzhou Medical College Institutional Review Board. Total RNA was extracted from cell lines or tissue samples with TRIzol reagent (Invitrogen), and the integrity was confirmed using spectrophotometry and formaldehyde/agarose gel electrophoresis. 10 μg of total RNA were dissolved in gel loading buffer II (Ambion, Austin, TX), heated at 95 °C for 3 min, loaded onto denaturing 15% Tris borate-EDTA-urea gels, and separated on a 15% denaturing urea-PAGE gel for 1 h and then transferred onto positively charged nylon membranes (GE Healthcare) followed by a cross-linking with UV irradiation. The RNA blots were prehybridized at 68 °C for 1 h using ULTRAhyb ultrasensitive hybridization buffer (Ambion) and subjected to hybridization with 3′-digoxigenin (DIG)-labeled locked nucleic acid probe for miR-1 or miR-206 (100 ng/ml) overnight at 42 °C. The locked nucleic acid-modified oligonucleotide probe was obtained from Exiqon (Vedbaek, Denmark). 100 pmol of the probe were DIG-labeled using DIG oligonucleotide 3′-end labeling kit (Roche Applied Science, Mannheim, Germany). Following hybridization, membranes were rinsed and then washed three times using a low stringency buffer (2× SSC and 0.1% SDS). Detection was performed using the DIG luminescent detection kit (Roche Applied Science) according to manufacturer's instructions. In brief, membranes were blocked in blocking buffer for 30 min and then incubated with alkaline phosphatase-conjugated anti-DIG antibody for 60 min followed by washing three times in washing buffer. After equilibration in detection buffer, blots were incubated with chemiluminescent substrate CDP-Star and exposed to a Kodak Biomax MR film (Eastman Kodak Co.). DIG-labeled U6 small nuclear RNA probe was used as an internal control. RD cells were plated at 3 × 103 cells/well in 96-well plates for each transfection. Transfections were performed using Lipofectamine 2000 (Invitrogen). For each well, 50 nm miR-1 or miR-206 precursor molecule or a negative control precursor miRNA was employed. For convenience, the miR-1 precursor or the miR-206 precursor is termed miR-1 or miR-206, respectively, following transfection throughout this report. Pre-miRTM miRNA precursor molecules (Ambion) are small, chemically modified double-stranded RNA molecules designed to mimic endogenous mature miRNAs once properly transfected and expressed by recipient cells. The negative control is a scrambled oligonucleotide that has been validated to not produce identifiable effects on known miRNA function (Ambion). After a 24-h culture, cell proliferation was assessed using the CellTiter 96 AQueous MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay (Promega, Madison, WI) according to the manufacturer's instructions. Briefly, the CellTiter 96 AQueous one solution reagent was added to each well and incubated at 37 °C for 3 h. Cell proliferation was assessed by measuring the absorbance at 490 nm using a microtiter plate reader (Molecular Devices, Sunnyvale, CA). RD cells were plated in 24-well plates and transfected with different miRNAs as described above. 24 h later, doxorubicin (0.1 μg/ml; Sigma) was added to each well prior to staining with Hoechst 33342 (5 μg/ml; Sigma) to visualize nuclear condensation and DNA fragmentation. After 20 min of staining at 25 °C, the cells were examined under fluorescence microscopy (Zeiss, Oberkochen, Germany). Apoptosis in RD cells was determined using the Caspase-Glo 3/7 assay kit according to the manufacturer's instructions (Promega). RD cells were first plated in triplicates in 96-well plates and transfected with different miRNAs as described above. Samples were then incubated with the caspase substrate for 2 h followed with measurements by a microtiter plate reader (Molecular Devices). RD cells were transfected with 50 nm miR-1 or miR-206 precursor molecule or a negative control. After 48 h, the cells were collected, washed with phosphate-buffered saline, and stained with propidium iodide using the BD Cycletest Plus DNA reagent kit (BD Biosciences). The stained cells (1 × 105) were then analyzed for DNA content with a flow cytometer (FACScaliber, BD Biosciences). The 3′-untranslated region (UTR) of human c-Met was amplified from human genomic DNA and individually cloned into the pMIR-REPORT vector (Ambion) by directional cloning. Seed regions were mutated to remove all complementarity to nucleotides 1–7 of miR-1/206 by using the QuikChange XL mutagenesis kit (Stratagene, La Jolla, CA). HEK-293 cells were co-transfected with 0.4 μg of firefly luciferase reporter vector and 0.02 μg of the control vector containing Renilla luciferase, pRL-SV40 (Promega), using Lipofectamine 2000 (Invitrogen) in 24-well plates. Each transfection was carried out in four wells. For each well, 50 nm miR-1 or miR-206 precursor molecule (Ambion) or a negative control precursor miRNA (Ambion) was co-transfected with the reporter constructs. Luciferase assays were performed 24 h after transfection using the Dual-Luciferase reporter assay system (Promega). Firefly luciferase activity was normalized to Renilla luciferase activity. RD cells were grown in DMEM containing 10% fetal bovine serum to ∼60% confluence and transfected with 50 nm miR-1 or miR-206 precursor molecule or a negative control. After 24 h, the cells were harvested by trypsinization and washed once with Hanks' balanced salt solution (Invitrogen). To measure cell migration, 8-mm pore size culture inserts (Transwell; Costar, High Wycombe, UK) were placed into the wells of 24-well culture plates, separating the upper and the lower chambers. In the lower of DMEM containing human hepatocyte growth factor 20 ng/ml) were HGF was purchased from 1 × cells were added to the upper After 24 h of at 37 °C with 5% the of cells that through the was by 10 under the using a and cell was by staining with and RD cells (1 × 105) were and grown in DMEM with 10% fetal bovine serum in plates for 24 h. After the cells were washed with phosphate-buffered and subjected to in a buffer 1 20 10 of protein μg and (GE Healthcare) were separated by 10% and then to The membranes were blocked with a buffer containing 5% in phosphate-buffered with 20 for 2 h and incubated overnight with antibody at °C. After a with phosphate-buffered containing the membranes were incubated with and with an enhanced detection kit dehydrogenase was used as a loading control. for total extracellular signal-regulated kinases 1 and 2 total total focal adhesion kinase and were from Cell and for c-Met, and were from The expression and control vector were purchased from CA). The was according to the manufacturer's instructions. RD cells were with miR-206, or negative mice of were used for RD cells × miR-1/206 or negative control were into the of All mice were after of tumor cells. size was with a and was using the following × × and according to the S. von R. Cancer Res. Google Scholar). All studies and procedures were approved by the Wenzhou Medical College and All were shown as the samples were analyzed using the was at To miRNA was in the of rhabdomyosarcoma we first miR-1/206 expression in normal skeletal muscle and rhabdomyosarcoma. blot was performed using DIG-labeled locked nucleic acid probe for miR-1 or miR-206 to its expression in tissue specimens and the rhabdomyosarcoma RD cell line. miR-1 and miR-206 were highly expressed in normal skeletal muscle In expression of miR-1/206 was decreased in RD cells To the expression of miR-1/206 in human rhabdomyosarcoma, total RNA from human rhabdomyosarcoma specimens was analyzed by with results from the RD miR-1/206 expression was suppressed or not detected in all samples and results that miR-1/206 expression is down-regulated in human rhabdomyosarcoma and a role of miR-1/206 in rhabdomyosarcoma development. miR-1/206 expression was down-regulated in rhabdomyosarcoma, we to its biological activity on cell RD cells were first transfected with the the miR-206 precursor or a negative control. Although transfection with the negative control not RD cell both miR-1 and miR-206 a significant of RD cell growth as with that of control the MTS assay was carried out to growth at after transfection. miR-1 or miR-206 transfected cells a of RD cell proliferation as with that of control a The decrease in cell was significant cells transfected with miR-1/206 and cells transfected with a negative control at day led to miR-206 led to To of cell proliferation, we the of cell using Hoechst RD cells showed and of DNA following Hoechst staining RD cells transfected with miR-1 or miR-206 a and in cell as with control. then investigated as a of cell by caspase activity. 3/7 activity was increased in miR-1/206 transfected cells in comparison with negative control after 48 h to the that miR-1/206 cell proliferation, these cells were found to have cell 48 h after cells were stained with propidium iodide and analyzed by flow transfected with miR-1 or miR-206 showed and in comparison with with transfection and with negative control of miR-1/206 down-regulated cell including and which are important for cell progression and In which the cell was up-regulated these results that miR-1/206 expression suppressed RD cell growth by and by cell Cell migration, a for and was assessed using a RD cells were first transfected with the miR-1/206 precursor or a control were on culture and the of cells to to the of the inserts was determined in the presence of shown in the was decreased cells with negative control for 10 for miR-206, 3 Therefore, of miR-1/206 in cell in to demonstrated a role for miR-1/206 in RD we attempted to the cellular mechanisms cell proliferation and migration. was for miR-1/206 target potential binding sites of miR-1/206 were predicted in the c-Met the predicted miR-1/206 target sites and miR-1/206, the seed sequence for is shown To the regulation of c-Met through the two predicted binding we amplified the c-Met sequence and it of the firefly luciferase region of a vector with the putative binding sites were as described of miR-1 or miR-206 in cells with the significant of luciferase activity as to negative control of the two binding using a vector the of miR-1 or miR-206 to regulate luciferase expression results demonstrated that c-Met was a potential target of To that miR-1/206 was responsible for the of c-Met, RD cells were transfected with the miR-1/206 molecule or a negative control. Western blot showed that c-Met expression was not affected by the transfection of a negative c-Met expression was transfected with miR-1 or miR-206

Through the amide formation between amine-functionalized polyhedral oligomeric silsesquioxane (POSS) and oxygen-containing groups (e.g., epoxy and carboxyl groups) in graphene oxide (GO), we have synthesized POSS-functionalized graphene nanosheets (POSS-graphene), which are highly soluble in various organic solvents attractive for multifunctional applications. Thin films from solution casting of the resultant POSS-graphene were found to show superhydrophobic properties with a water/air contact angle of ∼157°, while the superhydrophobic POSS-graphene powder could be used to construct liquid marbles. In addition, the POSS-graphene hybrids were also used as novel nanofillers to increase the glass transition temperature (Tg) and decompose temperature (Td) for polymers.

<h3>Importance</h3> Reducing myopia progression can reduce the risk of associated ocular pathologies. <h3>Objective</h3> To evaluate whether spectacle lenses with higher lenslet asphericity have a higher myopia control efficacy throughout 2 years. <h3>Design, Setting, and Participants</h3> This double-masked randomized clinical trial was conducted between July 2018 and October 2020 at the Eye Hospital of Wenzhou Medical University in Wenzhou, China. Children aged 8 to 13 years with a cycloplegic spherical equivalent refraction (SER) of −0.75 D to −4.75 D and astigmatism with less than −1.50 D were recruited. A data and safety monitoring committee reviewed findings from a planned interim analysis in 2019. <h3>Interventions</h3> Participants were randomly assigned in a 1:1:1 ratio to receive spectacle lenses with highly aspherical lenslets (HAL), spectacle lenses with slightly aspherical lenslets (SAL), or single-vision spectacle lenses (SVL). <h3>Main Outcome and Measures</h3> Two-year changes in SER and axial length and their differences between groups. <h3>Results</h3> Of 157 participants who completed each visit (mean [SD] age, 10.4 [1.2] years), 54 were analyzed in the HAL group, 53 in the SAL group, and 50 in the SVL group. Mean (SE) 2-year myopia progression in the SVL group was 1.46 (0.09) D. Compared with SVL, the mean (SE) change in SER was less for HAL (by 0.80 [0.11] D) and SAL (by 0.42 [0.11] D;<i>P</i> ≤ .001). The mean (SE) increase in axial length was 0.69 (0.04) mm for SVL. Compared with SVL, increase in axial length was slowed by a mean (SE) of 0.35 (0.05) mm for HAL and 0.18 (0.05) mm for SAL (<i>P</i> ≤ .001). Compared with SVL, for children who wore HAL at least 12 hours every day, the mean (SE) change in SER was slowed by 0.99 (0.12) D, and increase in axial length slowed by 0.41 (0.05) mm. <h3>Conclusions and Relevance</h3> In this study, HAL and SAL reduced the rate of myopia progression and axial elongation throughout 2 years, with higher efficacy for HAL. Longer wearing hours resulted in better myopia control efficacy for HAL. <h3>Trial Registration</h3> Chinese Clinical Trial Registry Identifier:ChiCTR1800017683

The formation of stable blood clots or hemostasis is essential to prevent major blood loss and death from excessive bleeding. However, the body's own coagulation process is not able to accomplish timely hemostasis without the assistance of hemostatic agents. For developing novel topical hemostatic agents, tissue adhesives and sealants, it is necessary to understand the coagulation process and the hemostasis mechanism of different materials. Among hemostatic materials, polysaccharides are naturally derived polymers having excellent biodegradable and biocompatible properties. This review provides an overview of polysaccharide-based hemostatic materials and agents, including their advantages and drawbacks in hemostatic applications. Furthermore, polysaccharide-based hemostatic materials with anti-microbial and healing functions are also introduced.