Publications by authors named "Yinghui Dong"

18 Publications

  • Page 1 of 1

Diagnostic performance of elastography for breast non-mass lesions: A systematic review and meta-analysis.

Eur J Radiol 2021 Oct 2;144:109991. Epub 2021 Oct 2.

Department of Ultrasound, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, Guangdong, China. Electronic address:

Purpose: This systematic review and meta-analysis aimed to evaluate the diagnostic performance of ultrasound elastography in the differentiation of benign and malignant breast non-mass lesions (NMLs).

Methods: PubMed, Cochrane Library, and Embase databases were searched for eligible studies up to end of June 2021. The diagnostic performance of elastography for NMLs was investigated using pooled sensitivity and specificity, likelihood ratio, diagnostic odds ratio (DOR), post-test probability, and the area under hierarchical summary receiver operating characteristic curve (HSROC).

Results: Eleven studies involving 812 NMLs (malignant 414) were included. The pooled sensitivity, specificity, DOR, positive likelihood ratio, and negative likelihood of elastography for the differentiation of benign and malignant breast NMLs were 79% (95 %CI: 71-85), 86% (95 %CI: 79-91), 23.32 (95 %CI: 13.38-40.66), 5.67 (95 %CI: 3.79-8.47), and 0.24 (95 %CI: 0.17-0.34), respectively. No significant publication bias existed. The area under the HSROC curve was 90% (95 %CI: 87-92). Fagan plots demonstrated good clinical utility. However, substantial heterogeneity existed. Country, measurement index, and number of lesions served as potential sources of heterogeneity.

Conclusions: The results of this study suggest that elastography has high diagnostic accuracy in differentiating between malignant and benign NMLs. Elastography can be a feasible and non-invasive tool for breast NMLs.
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http://dx.doi.org/10.1016/j.ejrad.2021.109991DOI Listing
October 2021

Vital Role of Gene in Ammonia Detoxification and the Association Between its SNPs and Ammonia Tolerance in .

Front Physiol 2021 5;12:664804. Epub 2021 May 5.

Ninghai Institute of Mariculture Breeding and Seed Industry, Zhejiang Wanli University, Ninghai, China.

Increasing evidence has revealed accumulated ammonia will cause adverse effects on the growth, reproduction, and survival of aquatic animals. As a marine benthic mollusk, the razor clam shows better growth and survival under high ammonia nitrogen environment. However, little is known about its adaptation mechanisms to high ammonia stress in an integrated mariculture system. In this study, we analyzed the association between the polymorphism of gene (), a key gene involved in ammonia nitrogen detoxification, and ammonia tolerance. The results showed that 26 and 22 single-nucleotide polymorphisms (SNPs) of in (denoted as ) were identified from two geographical populations, respectively. Among them, two SNPs (c.323T > C and c.620C > T) exhibited a significant and strong association with ammonia tolerance, suggesting that gene could serve as a potential genetic marker for molecular marker-assisted selection to increase survival rate and production of . To observe the histological morphology and explore the histocellular localization of Sc-GDH, by paraffin section and hematoxylin-eosin staining, the gills were divided into gill filament (contains columnar and flattened cells) and gill cilia, whereas hepatopancreas was made up of individual hepatocytes. The results of immunohistochemistry indicated that the columnar cells of gill filaments and the endothelial cells of hepatocytes were the major sites for Sc-GDH secretion. Under ammonia stress (180 mg/L), the expression levels of were extremely significantly downregulated at 24, 48, 72, and 96 h ( < 0.01) after RNA interference. Thus, we can speculate that gene may play an important role in the defense process against ammonia stress. Overall, these findings laid a foundation for further research on the adaptive mechanisms to ammonia-nitrogen tolerance for .
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http://dx.doi.org/10.3389/fphys.2021.664804DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8131826PMC
May 2021

Defense responses of sulfur dioxygenase to sulfide stress in the razor clam Sinonovacula constricta.

Genes Genomics 2021 May 15;43(5):513-522. Epub 2021 Mar 15.

Zhejiang Key Laboratory of Aquatic Germplasm Resources, Zhejiang Wanli University, Ningbo, Zhejiang, 315100, People's Republic of China.

Background: Sulfide is a well-known toxicant widely distributed in the culture environment. As a representative burrowing benthic bivalve, the razor clam Sinonovacula constricta is highly sulfide tolerant. Mitochondrial sulfide oxidation is an important way for sulfide detoxification, where sulfur dioxygenase (SDO) is the second key enzyme.

Objective: To investigate the mechanism of sulfide tolerance in S. constricta, the molecular characterization of its SDO (designated as ScSDO) was studied.

Methods: The cDNA sequence of ScSDO was cloned by RACE technique. The response of ScSDO in gills and livers of S. constricta was investigated during sulfide exposure (50, 150, and 300 μM sulfide) for 0, 3, 6, 12, 24, 48, 72, and 96 h by qRT-PCR. Moreover, the temporal expression of ScSDO protein in S. constricta gills after exposure to 150 μM sulfide was detected by Western blot. The subcellular location of ScSDO was identified by TargetP 1.1 prediction and Western Blot analysis.

Results: The full-length cDNA of ScSDO was 2914 bp, encoding a protein of 304 amino acids. The deduced ScSDO protein was highly conserved, containing the signature HXHXDH motif of the metallo-β-lactamase superfamily and two metal-binding sites, of which metal-binding site I is known to be the catalytically active center. Subcellular localization confirmed that ScSDO was located only in the mitochondria. Responding to the sulfide exposure, distinct time-dependent increases in ScSDO expression were detected at both mRNA and protein levels. Moreover, the gills exhibited a higher ScSDO expression level than the livers.

Conclusions: All of our results suggest that ScSDO plays an important role in mitochondrial sulfide oxidation during sulfide stress, making S. constricta highly sulfide tolerant. In addition, as a respiratory tissue, the gills play a more critical role in sulfide detoxification.
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http://dx.doi.org/10.1007/s13258-021-01077-0DOI Listing
May 2021

The razor clam Sinonovacula constricta uses the strategy of conversion of toxic ammonia to glutamine in response to high environmental ammonia exposure.

Mol Biol Rep 2020 Dec 27;47(12):9579-9593. Epub 2020 Nov 27.

Zhejiang Key Laboratory of Aquatic Germplasm Resource, College of Biological & Environmental Sciences, Zhejiang Wanli University, Ningbo, 315100, People's Republic of China.

High ammonia can inhibit the survival and growth, and even cause mortality of razor clam (S. constricta). The accumulation of ammonia to lethal concentrations in some invertebrates may be partially prevented by converting some of the ammonia into glutamine (Gln). Glutamine dehydrogenase (GDH) and glutamine synthetase (GS) have been widely implicated a central role in response to ammonia stress. However, the molecular and physiological response of GDH and GS to ammonia alterations has not yet been determined in clams. To investigate the possible participatory role of GDH and GS genes in altered ammonia conditions, we have cloned their gene sequences and examined the mRNA expression and western blotting under ammonia exposure in S. constricta (ScGDH and ScGS), and detected the levels of GS and GDH, and the content of glutamate (Glu) and Gln. The full-length cDNA of ScGDH was 3924 bp, with a 1629 bp open reading frame (ORF) encoding a 542 amino-acid polypeptide. The complete cDNA sequence for ScGS had 2739 bp with an ORF of 1110 bp encoding 369 amino acids. To investigate ammonia detoxification strategies, the clams were exposed to ammonia for 96 h at four different concentrations (0, 100, 140, and 180 mg/L). Exposure to ammonia resulted in a significant increase of glutamate concentration and Gln in the haemocytes. GDH activity, GDH relative mRNA and protein expression, GS activity, GS relative mRNA and protein expression increased significantly and showed a pronounced time and dosage interaction in the liver. The results suggested that the protective strategies of Gln formation existed in S. constricta, which could convert ammonia to non- or less toxic nitrogenous compounds on the exposure of ammonia. Glutamate content in the haemocytes increased significantly, which is to ensure sufficient Glu to meet the needs for GS to catalyze the conversion of ammonia to Gln. We proposed that the induction of Glu synthesis-related genes and the subsequent formation of the active protein occurred in preparation for the increased capacity of the body to convert ammonia, into Gln. The results of this study suggested that GDH and GS play an important role in the synthesis of Gln, emphasizing, the protective strategies of Gln formation in S. constricta convert ammonia to nontoxic or less toxic nitrogenous compounds upon exposure to ammonia.
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http://dx.doi.org/10.1007/s11033-020-06018-wDOI Listing
December 2020

A Potential Role of Bone Morphogenetic Protein 7 in Shell Formation and Growth in the Razor Clam .

Front Physiol 2020 28;11:1059. Epub 2020 Aug 28.

Zhejiang Key Laboratory of Aquatic Germplasm Resources, College of Biological & Environmental Sciences, Zhejiang Wanli University, Ningbo, China.

Bone morphogenetic proteins (BMPs) not only play essential roles in bone development but also are involved in embryonic growth, organogenesis cell proliferation and differentiation. However, the previous studies on the functions of shellfish BMPs genes are still very limited. To better understand its molecular structure and biological function, BMP7 of the razor clam (7) was cloned and characterized in this study. The full length of is 2252 bp, including an open reading frame (ORF) of 1257 bp encoding 418 amino acids. The protein sequence included a signal peptide (1-32 aa), a prodomain (38-270 aa) and a TGF-β domain (317-418 aa). The quantitative expression of eleven adult tissues showed that 7 was significantly higher expressed in the gill, foot, and mantle ( < 0.05), but lower in hemocytes and hepatopancreas. In the early development stages, low expression was detected in the stages of unfertilized mature eggs, fertilized eggs, 4-cell embryos, blastula, gastrulae, whereas it increased after the stage of trochophore and demonstrated the highest expression in umbo larvae ( < 0.01). In shell repair experiment, 7 showed increasing expression level after 12 h. The higher expression of 7 was detected while Ca concentration was reduced in seawater. After inhibiting 7 expression using RNA interference (RNAi) technology, expression of 7 mRNA and protein were significantly down-regulated ( < 0.05) in the central zone of mantle (nacre formation related tissue) and the pallial zone of mantle (prismatic layer formation related tissue). Association analysis identified two shared SNPs in exon of 7 gene from 246 individuals of two groups. These results indicated that 7 might be involved in shell formation and growth. These results would contribute to clarify the role of 7 in the regulation of growth and shell formation, and provide growth-related markers for molecular marker assisted breeding of this species.
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http://dx.doi.org/10.3389/fphys.2020.01059DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7485270PMC
August 2020

The Chromosome-Level Genome Assembly and Comprehensive Transcriptomes of the Razor Clam ().

Front Genet 2020 7;11:664. Epub 2020 Jul 7.

Key Laboratory of Aquatic Germplasm Resource of Zhejiang, College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo, China.

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http://dx.doi.org/10.3389/fgene.2020.00664DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7358530PMC
July 2020

Catalytic pyrolysis and liquefaction behavior of microalgae for bio-oil production.

Bioresour Technol 2020 Mar 24;300:122665. Epub 2019 Dec 24.

Institute of Tribology, School of Mechanical Engineering, Hefei University of Technology, Hefei 230009, China.

Microalgae bio-oil production is related to the sustainable use of world energy in the future. In the present work, catalytic pyrolysis and liquefaction behavior of microalgae for bio-oil production were investigated. The results show that the rare earth compounds as catalysts contributed to significantly accelerating the pyrolysis of microalgae via reducing the activation energy of pyrolysis process. Ce(II)/HZSM-5 presented the optimal catalytic pyrolysis and liquefaction effects by helping cut the microalgae molecule chains. The maximum bio-oil yield amounted to 49.71 wt% at the catalyst concentration of 5 wt%. The chemical components of the Spirulina bio-oil were composed of carboxylic acids, ketones, olefins, amides, ethers, esters, and partially cyclic N-containing compounds. Although the combustion performances of the Spirulina bio-oil are worse than those of the diesel fuel, it is superior to the reported rice husk bio-oil, suggesting a promising potential application prospect.
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http://dx.doi.org/10.1016/j.biortech.2019.122665DOI Listing
March 2020

Polymorphisms of LAP3 gene and their association with the growth traits in the razor clam Sinonovacula constricta.

Mol Biol Rep 2020 Feb 19;47(2):1257-1264. Epub 2019 Dec 19.

Key Laboratory of Aquatic Germplasm Resource of Zhejiang, College of Biological & Environmental Sciences, Zhejiang Wanli University, 8 South Qianhu Road, Ningbo, 315100, Zhejiang, China.

Leucine aminopeptidase 3 (LAP3) is an important proteolytic enzyme that catalyzes the hydrolysis of leucine residues from the amino termini of protein or peptide substrates and plays a critical role in protein metabolism and growth. In the present study, we investigated the full-length cDNA sequence of the LAP3 gene in Sinonovacula constricta (ScLAP3) using expressed sequence tags and rapid amplification of cDNA ends. The full-length ScLAP3 cDNA was 2885 bp, with a 1560 bp open reading frame encoding 519 amino acids. Sequence analysis revealed that ScLAP3 shared 70.9% identity with LAP3 from the blood clam Tegillarca granosa and 62.0-68.0% with other species. ScLAP3 was expressed in all six tested tissues, with significantly higher expression levels in the foot compared with mantle, adductor muscle, liver, gills, and siphon tissues in adults (P < 0.01). In the eight developmental stages, ScLAP3 expression gradually increased, with significantly higher levels in D-shaped larvae compared with other developmental stages (P < 0.01), suggesting that it may be involved in the formation of certain organs during early development. Association analysis identified three shared single nucleotide polymorphisms (SNPs), c.1073A > G, c.1139C > T and c.1154A > G in exons of ScLAP3 gene from 177 individuals of two groups, one selective strain and one wild population, which had significant effects on growth traits of S. constricta. The results provided candidate genetic markers to assist selective breeding of razor clams toward improved growth.
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http://dx.doi.org/10.1007/s11033-019-05231-6DOI Listing
February 2020

The Highly Conserved Barley Powdery Mildew Effector Confers Susceptibility to Biotrophic and Necrotrophic Pathogens in Wheat.

Int J Mol Sci 2019 Sep 6;20(18). Epub 2019 Sep 6.

Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou 466001, China.

Effector proteins secreted by plant pathogens play important roles in promoting colonization. effector candidate () 1019, a highly conserved metalloprotease of f. sp. (), is essential for fungal haustorium formation, and silencing significantly reduces virulence. In this study, we found that homologs in f. sp. () and var. () have complete sequence identity with those in , prompting us to investigate their functions. Transcript levels of were abundantly induced concomitant with haustorium formation in and necrosis development in -infected plants. overexpression considerably increased wheat susceptibility to and , whereas silencing this gene using host-induced gene silencing significantly enhanced wheat resistance to and , which was associated with hydrogen peroxide accumulation, cell death, and pathogenesis-related gene expression. Additionally, we found that the full and partial sequences of can trigger cell death in leaves. These results indicate that and can utilize as a virulence effector to promote plant colonization, and thus these genes represent promising new targets in breeding wheat cultivars with broad-spectrum resistance.
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http://dx.doi.org/10.3390/ijms20184376DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6770355PMC
September 2019

Transcriptomics analysis revealing candidate genes and networks for sex differentiation of yesso scallop (Patinopecten yessoensis).

BMC Genomics 2019 Aug 23;20(1):671. Epub 2019 Aug 23.

Key Laboratory for Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Science, Qingdao, China.

Background: The Yesso scallop, Patinopecten (Mizuhopecten) yessoensis, is a commercially important bivalve in the coastal countries of Northeast Asia. It has complex modes of sex differentiation, but knowledge of the mechanisms underlying this sex determination and differentiation is limited.

Results: In this study, the gonad tissues from females and males at three developmental stages were used to investigate candidate genes and networks for sex differentiation via RNA-Req. A total of 901,980,606 high quality clean reads were obtained from 18 libraries, of which 417 expressed male-specific genes and 754 expressed female-specific genes. Totally, 10,074 genes differentially expressed in females and males were identified. Weighted gene co-expression network analysis (WGCNA) revealed that turquoise and green gene modules were significantly positively correlated with male gonads, while coral1 and black modules were significantly associated with female gonads. The most important gene for sex determination and differentiation was Pydmrt 1, which was the only gene discovered that determined the male sex phenotype during early gonadal differentiation. Enrichment analyses of GO terms and KEGG pathways revealed that genes involved in metabolism, genetic and environmental information processes or pathways are sex-biased. Forty-nine genes in the five modules involved in sex differentiation or determination were identified and selected to construct a gene co-expression network and a hypothesized sex differentiation pathway.

Conclusions: The current study focused on screening genes of sex differentiation in Yesso scallop, highlighting the potential regulatory mechanisms of gonadal development in P. yessoensis. Our data suggested that WCGNA can facilitate identification of key genes for sex differentiation and determination. Using this method, a hypothesized P. yessoensis sex determination and differentiation pathway was constructed. In this pathway, Pydmrt 1 may have a leading function.
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http://dx.doi.org/10.1186/s12864-019-6021-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6708199PMC
August 2019

MiR-122 inhibits epithelial mesenchymal transition by regulating P4HA1 in ovarian cancer cells.

Cell Biol Int 2018 Nov 16;42(11):1564-1574. Epub 2018 Sep 16.

Department of Gynecology, The Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, People's Republic of China.

Ovarian cancer is one of the most common gyneacologic malignancies, with high morbidity and high mortality. Hsa-miR-122-5p (miR-122) has been reported with tumor-suppressing roles in various cancers. In this study, miR-122 was overexpressed in ovarian cancer cells, and phenotypic experiments demonstrated that miR-122 inhibited migration and invasion in SKOV3 and OVCAR3 cells. MiR-122 also suppressed epithelial mesenchymal transition (EMT), evidenced by expression changes of E-cadherin, vimentin, matrix metalloproteinase (MMP)2, and MMP14. Prolyl-4-hydroxylase subunit alpha-1 (P4HA1) was identified as a target of miR-122, and downregulated by miR-122. MiR-122-induced the elevation of migration, invasion, and EMT were recovered by P4HA1. Additionally, miR-122 restrained the tumor metastasis of SKOV3 cells in peritoneal cavity of nude mice. In summary, we demonstrated that miR-122 inhibited migration, invasion, EMT, and metastasis in peritoneal cavity of ovarian cancer cells by targeting P4HA1 for the first time, which shed lights on the discovery of miR-122 and P4HA1 as possible potential diagnostic markers and therapeutic targets for ovarian cancer.
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http://dx.doi.org/10.1002/cbin.11052DOI Listing
November 2018

Genetic analysis assessed by microsatellites for a diallel mating design of two geographical stocks of the blood clam Tegillarca granosa.

Genes Genomics 2018 04 25;40(4):373-379. Epub 2017 Nov 25.

Zhejiang Key Laboratory of Aquatic Germplasm Resources, College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo, People's Republic of China.

To determine the potential for productive efficiency and genetic improvement in the blood clam Tegillarca granosa, four offspring populations (ZZ, ZK, KZ and KK) were produced from a diallel mating of two different geographical stocks (Z and K). The levels of genetic diversity and population structures of four populations were analyzed using 14 polymorphic microsatellites. The results showed that the mean observed heterozygosities (Ho) of reciprocal cross populations (ZK and KZ) was higher than those of pure populations(ZZ and KK). The largest values of genetic differentiation coefficient (F = 0.067) and Nei's unbiased genetic distance (Dc = 0.263) were between ZK and KZ, and the smallest (F = 0.020, Dc = 0.116) were between ZZ and KK, which revealed that the largest genetic divergence was between the two reciprocal cross populations and the smallest was between two pure populations. This study demonstrated that the reciprocal cross populations of T. granosa had an extensive genetic difference and improvement, which may be advantageous for future breeding studies.
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http://dx.doi.org/10.1007/s13258-017-0636-2DOI Listing
April 2018

Two I84 family protease inhibitors from Chinese razor clams Sinonovacula constricta expressed in response to environmental challenges.

Fish Shellfish Immunol 2018 Apr 8;75:149-157. Epub 2018 Feb 8.

Key Laboratory of Aquatic Germplasm Resource of Zhejiang, Zhejiang Wanli University, Ningbo, Zhejiang 315100, China. Electronic address:

Protease inhibitors play critical roles in numerous biological processes including host defense in all multicellular organisms. Eighty three evolutionary families of protease inhibitors are currently accommodated in the MEROPS database and the I84 family currently consists of 3 novel serine protease inhibitors from the eastern oyster Crassostrea virginica. In this study, we identified 2 new I84 family members from the Chinese razor clam Sinonovacula constricta, scSI-1 and scSI-2, using cDNA cloning and sequencing. The scSI-1 cDNA consisted of 494 bp with a 282 bp ORF encoding a 93-amino acid polypeptide that was predicted to have a 19-amino acid signal peptide and a 74-residue mature protein with a calculated molecular mass of 8248.5 Da. The scSI-2 cDNA was 490 bp long with a 273 bp ORF encoding a 90-amino acid polypeptide that was predicted to have an 18-amino acid signal peptide and a 72-residue nature protein with a calculated molecular mass of 7528.4 Da. ScSI-1 and scSI-2 shared high sequence similarity with the 3 known members of I84 family and both expressed primarily in the clam digestive glands. Protease inhibitory activity in the clam plasma also exhibited the signature kinetic characteristics of the I84 members from the oyster. In addition, levels of scSI-1 and scSI-2 gene expression in digestive glands and the protease inhibitory activity in plasma elevated significantly in clams challenged by bacterial injections and Vibrio harveyi was more effective than Staphylococcus epidermidis in inducing the gene expression and plasma protease inhibitory activity. Moreover, drastic changes of salinity and temperature also caused significant changes in the gene expression and plasma activity. These results indicated that scSI-1 and scSI-2 represented 2 new members of the I84 family and they likely play a role in clam host defense against infections and in reactions against physiochemical stressors.
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http://dx.doi.org/10.1016/j.fsi.2018.02.016DOI Listing
April 2018

Integration of Next Generation Sequencing and EPR Analysis to Uncover Molecular Mechanism Underlying Shell Color Variation in Scallops.

PLoS One 2016 26;11(8):e0161876. Epub 2016 Aug 26.

Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, China.

The Yesso scallop Patinopecten yessoensis displays polymorphism in shell colors, which is of great interest for the scallop industry. To identify genes involved in the shell coloration, in the present study, we investigate the transcriptome differences by Illumina digital gene expression (DGE) analysis in two extreme color phenotypes, Red and White. Illumina sequencing yields a total of 62,715,364 clean sequence reads, and more than 85% reads are mapped into our previously sequenced transcriptome. There are 25 significantly differentially expressed genes between Red and White scallops. EPR (Electron paramagnetic resonance) analysis has identified EPR spectra of pheomelanin and eumelanin in the red shells, but not in the white shells. Compared to the Red scallops, the White scallops have relatively higher mRNA expression in tyrosinase genes, but lower expression in other melanogensis-associated genes. Meantime, the relatively lower tyrosinase protein and decreased tyrosinase activity in White scallops are suggested to be associated with the lack of melanin in the white shells. Our findings highlight the functional roles of melanogensis-associated genes in the melanization process of scallop shells, and shed new lights on the transcriptional and post-transcriptional mechanisms in the regulation of tyrosinase activity during the process of melanin synthesis. The present results will assist our molecular understanding of melanin synthesis underlying shell color polymorphism in scallops, as well as other bivalves, and also help the color-based breeding in shellfish aquaculture.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0161876PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5001709PMC
August 2017

Ferritin has an important immune function in the ark shell Scapharca broughtonii.

Dev Comp Immunol 2016 Jun 25;59:15-24. Epub 2015 Dec 25.

Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, PR China.

Ferritin, the principle cytosolic iron storage protein in the majority of living organisms, has important roles during immune process in invertebrates. Detailed information about ferritin in the ark shell Scapharca broughtonii, however, has been very limited. In this study, full-length ferritin (termed SbFer) was cloned by the rapid amplication of cDNA ends (RACE) method based upon the sequence from the transcriptome library. The cDNA contained a 182 bp 5'-untranslated region, a 519 bp open reading frame encoding a polypeptide of 172 amino acids, a 229 bp 3'-untranslated region, and three introns (902, 373 and 402 bp) embedded in four exons. There was an iron response element (IRE) in the 5'-untranslated region. The deduced amino acid sequence of SbFer possessed many characteristics of vertebrate H type ferritin, shared 63%-91% identity with mollusks and greater identity with vertebrate H type ferritin compared to the L type. The SbFer gene expression pattern examined by quantitative real-time PCR showed ferritin mRNA was expressed in all ark shell tissues examined. The highest levels of expression were found in hemocytes with decreasing levels of expression in foot, mantle, gill, adductor muscle and hepatopancreas. A challenge with Vibrio anguillarum resulted in time-dependent significant upregulation of SbFer mRNA, indicating SbFer participated actively in the bacterial defense process. Further analysis of the antibacterial activity indicated recombinant SbFer could function as an immune antibacterial agent to both Gram-positive and Gram-negative bacteria. Taken together, these results suggested strongly that ferritin of the ark shell is involved in immune defense against microbial infection and it is a constitutive and inducible acute-phase protein.
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http://dx.doi.org/10.1016/j.dci.2015.12.010DOI Listing
June 2016

Identification and comparative analysis of the Tegillarca granosa haemocytes microRNA transcriptome in response to Cd using a deep sequencing approach.

PLoS One 2014 1;9(4):e93619. Epub 2014 Apr 1.

Zhejiang Key Laboratory of Aquatic Germplasm Resources, Zhejiang Wanli University, Ningbo, Zhejiang, China.

Background: MicroRNAs (miRNAs) are endogenous non-coding small RNAs (sRNAs) that can base pair with their target mRNAs, which represses their translation or induces their degradation in various biological processes. To identify miRNAs regulated by heavy metal stress, we constructed two sRNA libraries for the blood clam Tegillarca granosa: one for organisms exposed to toxic levels of cadmium (Cd) and one for a control group.

Results: Sequencing of the two libraries and subsequent analysis revealed 215 conserved and 39 new miRNAs. Most of the new miRNAs in T. granosa were up- or down-regulated in response to Cd exposure. There were significant differences in expression between the Cd and control groups for 16 miRNAs. Of these, five miRNAs were significantly up-regulated and 11 were significantly down-regulated in the Cd stress library. Potential targets were predicted for the 16 differential miRNAs in pre-miRNAs identified according to sequence homology. Some of the predicted miRNA targets are associated with regulation of the response to stress induced by heavy metals. Five differentially expressed miRNAs (Tgr-nmiR-8, Tgr-nmiR-21, Tgr-miR-2a, Tgr-miR-10a-5p, and Tgr-miR-184b) were validated by qRT-PCR.

Conclusion: Our study is the first large-scale identification of miRNAs in T. granosa haemocytes. Our findings suggest that some miRNAs and their target genes and pathways may play critical roles in the responses of this species to environmental heavy metal stresses.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0093619PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3972184PMC
June 2015

Polymorphism of the multiple hemoglobins in blood clam Tegillarca granosa and its association with disease resistance to Vibrio parahaemolyticus.

Fish Shellfish Immunol 2013 May 5;34(5):1320-4. Epub 2013 Mar 5.

Zhejiang Key Lab of Aquatic Germplasm Resource, Zhejiang Wanli University, 8 South Qianhu Road, Ningbo, Zhejiang 315100, PR China.

Hemoglobin (Hb) is the major protein component of erythrocytes in animals with red blood, but it can serve additional functions beyond the transport of oxygen. In this study, we identified polymorphism in the blood clam Tegillarca granosa Hb (Tg-Hb) genes and investigated the association of this polymorphism with resistance/susceptibility to Vibrio parahaemolyticus. Analysis of the 540 sequences revealed 28 SNPs in the coding region of three Tg-Hbs, corresponding to about one SNP per 48 bp. Three SNPS: HbIIA-E2-146, HbIIB-E2-23, HbIIB-E2-121 showed a significant association with resistance/susceptibility to V. parahaemolyticus (P < 0.05). To further demonstrate that three significant SNPs of Tg-Hbs is associated with resistance of clams to V. parahaemolyticus, SNPs were genotyped in V. parahaemolyticus resistant strain clams and the wild base population from which this strain was derived. The results indicated that the nonsynonymous mutation T allele at HbIIA-E2-146 and A allele at HbIIB-E2-23 are associated with V. parahaemolyticus resistance in the blood clam, and its association with disease resistance may be due to its cause changes in amino acid sequences to a functional polymorphism. Together with previous bacterial challenge study, these results provides direct evidence that variation at HbIIA-E2-146 and HbIIB-E2-23 are associated with disease resistance in the blood clam, and these two polymorphic loci could be potential gene markers for the future molecular selection of strains that are resistant to diseases caused by V. parahaemolyticus.
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http://dx.doi.org/10.1016/j.fsi.2013.02.022DOI Listing
May 2013

Real-time elastography for diagnosis of liver fibrosis in chronic hepatitis B.

J Ultrasound Med 2012 Jul;31(7):1053-60

Department of Ultrasound, Shengjing Hospital of China Medical University, 36 Sanhao St 110004, Shenyang, Liaoning, China.

Objectives: The purpose of this study was to prospectively investigate the value of real-time ultrasound elastography for diagnosis of liver fibrosis in patients with chronic hepatitis B and to correlate the elastographic findings with histologic stages of liver fibrosis and blood parameters.

Methods: Liver biopsies, blood testing, and real-time elastography were performed in 71 patients with chronic viral hepatitis B and liver cirrhosis. The ratio of the elastic strain of liver tissue to that of muscle tissue was determined and correlated with the histologic fibrosis stages and laboratory examination results.

Results: There was a highly negative correlation between the elastic strain ratio and the histologic fibrosis stage (Spearman r = -0.702; P < .001). There was a high correlation observed between a decreasing elastic strain ratio and an increasing fibrosis stage. With substantial liver fibrosis (Scheuer score ≥ S2) and cirrhosis (S4) as diagnostic criteria, the areas under the receiver operating characteristic curves (AUCs) of the elastic strain ratios were 0.863 and 0.797, respectively. The AUC for substantial fibrosis was higher than the AUC for the blood parameters used to diagnose substantial liver fibrosis. Elastic strain ratio cutoff values of 1.10 and 0.60 were identified as diagnostic of substantial fibrosis and cirrhosis, respectively, with sensitivities of 77.8% and 50.0%, respectively, and specificities of 80.0% and 96.7%.

Conclusions: Real-time elastography is a new clinically promising and noninvasive method for quantitative assessment of liver fibrosis.
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http://dx.doi.org/10.7863/jum.2012.31.7.1053DOI Listing
July 2012
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