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To estimate the up-to-date prevalence of chronic kidney disease among the health check-up population in economically developed areas of China using estimated glomerular filtration rate, urinary albumin creatinine ratio, and kidney ultrasound. Healthcare data from 38,093 subjects in 10 megalopolises of China who had an annual health check-up in 2021 were used. The overall and stratified prevalence of chronic kidney disease by sex, age, region and comorbidity group was reported. The association between chronic kidney disease and covariates of demographics, and comorbidities were analyzed in the multivariable-adjusted logistic regression model. A total of 3837 CKD cases were detected meeting any of the three CKD diagnostic criteria, with a crude prevalence of 10.1% in the study population. Using one criterion of decreased glomerular filtration rate, albuminuria and kidney structural abnormalities alone detected 204 (5.3%), 3289 (85.7%) and 563 (14.7%) cases, respectively. The addition of kidney ultrasound detected 427 (11.1%) structural abnormality cases without decreased GFR and albuminuria. The most common abnormalities were renal masses, hydronephrosis due to obstruction and congenital anomalies of kidney and urinary tract. Female, older age, low city-tier, hypertension, diabetes, obesity, hypertriglyceridemia as well as early disease stages such as pre-hypertension, impaired fasting glucose and overweight were significantly associated with chronic kidney disease. Kidney ultrasound helps to amplify the detection of CKD patients, which is a supplement to kidney function and urine protein. [ABSTRACT FROM AUTHOR]

As China's environmental problems have been thrust into the limelight of both domestic and international media, the Chinese government is eager to present a positive country image in terms of environmental planning and protection. Against this backdrop, this study aims to investigate how a "Green China" image is constructed through state-sanctioned eco-documentaries. Drawing on the Attitude System in Appraisal theory and multimodal discourse analysis, the study develops an analytical framework to investigate country image as semantic categories, as well as how it is constructed through multimodal resources. The analysis shows that these documentaries create an eco-friendly country image through: (1) highlighting the significance and desirability of the Chinese landscape (appreciation), (2) constructing a positive image of Chinese social actors in terms of environmental protection (judgment), and (3) depicting non-human characters' happiness and security of living in China (affect). These positive attitudes reflect and constitute China's green image branding practices shaped by the entangled forces of the Party-state's rejuvenation of traditional culture, its development of globally competitive green technologies, and its promotion of participatory environmental governance. [ABSTRACT FROM AUTHOR]

As China's national digital strategy proceeds into its implementation phase, there is an urgent need to further deepen the integration and synergy between technology and education to take the full advantages of digital education in this new era. Technological innovation and digital transformation can generate new momentum in high-quality education development by causing a major shift in educational paradigms and practice models. While the micro-level implementation of digital transformation on education has already been extensively discussed in the educational sector, the efforts in answering the same question from a macro-level perspective is currently lacking. The present study addresses this gap by discussing the implementation of digital transformation on education through the lens of diffusion of innovation theory. Specifically, we examine how the theory can provide unique insights on studying the developmental status, process, and outcome of digital transformation on education. As the application of diffusion of innovation theory in this topic, the study will contribute to the development of China's high-quality education system by providing theoretical guidance on the advancement of digital transformation on education under the overarching goal of education modernization. [ABSTRACT FROM AUTHOR]

In the digital age, cities around the world are mobilizing various symbolic resources to rebrand their images through social media. Against this background, this study investigates how Xi'an, a second-tier developing city in China, constructs its digitalized urban imaginary using the popular social media platform of TikTok. A semiotic framework is developed to model Xi'an's urban imaginary as evaluative attributes and to elucidate how they are constructed through linguistic and visual resources in short videos on TikTok. The analysis of 294 videos shows that Xi'an highlights its dual identity as a modern metropolis and a historical city. The modern metropolis image is characterized by the personification of Xi'an as a stylish, young, popular, and international microcelebrity; the historical city image is constructed through recreating the Great Tang dynasty and revitalizing local folk art. The characteristics of city branding discourse reflect China's wanghong economy, urban policies and the affordances of social media. [ABSTRACT FROM AUTHOR]

Mud crab (Scylla paramamosain) is one of the most economically-important marine crabs in China. However, research on mechanisms of reproductive regulation is not sufficient. Vitellogenesis-inhibiting hormone (VIH) is a member of the crustacean hyperglycemia hormones (CHH) family, which plays an essential role in the regulation of gonadal development and maturation in crustaceans, and current studies on the regulation of Vih transcription in crabs are relatively rare. Our previous studies on the transcriptional regulation of mud crab Vih (SpVih) have proved that the binding site of Oct4/Sox9 transcription factor may be the key region for positively regulating the expression of SpVih. In this study, the electrophoretic mobility shift assay (EMSA) experiment confirmed that the nuclear protein extracted from the eyestalk could bind to the key region of SpVih promoter, and these specific bindings were dependent on the presence of Oct4/Sox9 binding sites. Two specific binding complex bands were detected in the supershift group of EMSA supershift experiments by Oct4 and Sox9 antibodies, further confirming the specific recognition of these two transcription factors on the key regulatory region of SpVih. In vitro, Oct4 and Sox9 gene overexpression vectors and SpVih core promoter fragment vector were constructed and co-transfected into HEK293T cells. As a result, SpVih activity increased with the concentration of transcription factors. In vivo , when Oct4 and Sox9 dsRNA were injected into the eyestalks of mud crab, respectively, the expression level of SpVih decreased significantly after interference with Oct4 or Sox9 , and the expression level of SpVtg in the ovary and hepatopancreatic increased. Both in vitro and in vivo experiments showed that Oct4 and Sox9 had a positive regulatory effect on SpVih. The GST pull-down experiment was carried out by purified Oct4 and Sox9 proteins, and the results showed that there was an interaction between them. It was speculated that they regulated the expression of SpVih through the interaction. [ABSTRACT FROM AUTHOR]

Abstract: Accounting for 7% of energy consumption from all industrial sectors and 15% of total greenhouse gas (GHG) emissions in China in 2009, the cement industry should be one of the main sectors to implement low carbon development. However, a detailed study on presenting its GHG emission trajectory and identifying the key driving factors has yet to be done. Under such a circumstance, this paper first presents a GHG emission inventory of the cement industry in China and then identifies the main driving factors that influence changes of GHG emissions in the cement industry by adopting LMDI method. Our results show that the total GHG emission in 2009 was 1095.1 Mt carbon dioxide equivalent (CO2e), of which 592.8 Mt CO2e was process-related and 502.3 Mt CO2e was energy-related. The major factors responsible for the increase of GHG emissions during 2005–2009 include cement production activity effect and clinker production activity effect, while energy intensity effect played a positive role to offset total GHG emissions. Finally, we present our policy implications and provide possible technical solutions for reducing GHG emissions of the cement industry in China. [Copyright &y& Elsevier]

Background: The effects of diverse stresses ultimately alter the structures and functions of proteins. As molecular chaperones, heat shock proteins (HSPs) are a group of highly conserved proteins that help in the refolding of misfolded proteins and the elimination of irreversibly damaged proteins. They are mediated by a family of transcription factors called heat shock factors (HSFs). The small abalone Haliotis diversicolor is a species naturally distributed along the southern coast of China. In this study, the expression of HdHSF1 was inhibited by RNAi in hemocytes in order to further elucidate the regulatory roles of HdHSF1 on heat shock responsive genes in abalone. Meanwhile, to understand the transcriptional regulation of the HdHSF1 gene, the 5′-upstream regulatory region of HdHSF1 was characterized, and the relative promoter activity was examined by dual-luciferase reporter gene assay system in HEK293T cell lines. Results: After the inhibition of the H. diversicolor HSF1 gene (HdHSF1) by dsRNA (double-stranded RNA), the expression of most heat shock related-genes was down-regulated (p < 0.05). It indicated the importance of HdHSF1 in the heat shock response of H. diversicolor. Meanwhile, 5′-flanking region sequence (2633 bp) of the HdHSF1 gene was cloned; it contained a putative core promoter region, TATA box, CAAT box, CpG island, and many transcription elements. In HEK293T cells, the 5′-flanking region sequence can drive expression of the enhanced green fluorescent protein (EGFP), proving its promoter function. Exposure of cells to the high-temperature (39 °C and 42 °C) resulted in the activation of HdHSF1 promoter activity, which may explain why the expression of the HdHSF1 gene participates in heat shock response. Luciferase activity of different recombinant plasmids, which contained different truncated promoter fragments of the HdHSF1 gene in HEK293T cells, revealed the possible active regions of the promoter. To further identify the binding site of the critical transcription factor in the region, an expression vector with the site-directed mutation was constructed. After being mutated on the GATA-1 binding site, we found that the luciferase activity was significantly increased, which suggested that the GATA-1 binding site has a certain weakening effect on the activity of the HdHSF1 promoter. Conclusions: These findings suggest that GATA-1 may be one of the transcription factors of HdHSF1, and a possible signaling pathway mediated by HdHSF1 may exist in H. diversicolor to counteract the adverse effects of heat shock stress. [ABSTRACT FROM AUTHOR]

Mud crab (Scylla paramamosain) has become an important mariculture crab along the southeast coast of China due to its strong adaptability, delicious taste, and rich nutrition. Several vertebrate steroid hormones and their synthesis-related genes and receptors have been found in crustaceans, but there are few reports on their synthesis process and mechanism. 3-beta-hydroxysteroid dehydrogenase (HSD3B) is a member of the Short-chain Dehydrogenase/Reductase (SDR) family, and an indispensable protein in vertebrates' steroid hormone synthesis pathway. In this study, the SpHsd3b gene sequence was obtained from the transcriptome data of S. paramamosain , and its full-length open reading frame (ORF) was cloned. The spatial and temporal expression pattern of SpHsd3b was performed by quantitative real-time PCR (qRT-PCR). SpHsd3b dsRNA interference (RNAi) and HSD3B inhibitor (trilostane) were used to analyze the function of Sp HSD3B. The results showed that the SpHsd3b gene has an 1113 bp ORF encoding 370 amino acids with a 3β-HSD domain. Sp HSD3B has lower homology with HSD3B of vertebrates and higher homology with HSD3B of crustaceans. SpHsd3b was expressed in all examined tissues in mature crabs, and its expression was significantly higher in the testes than in the ovaries. SpHsd3b expression level was highest in the middle stage of testicular development, while its expression was higher in the early and middle stages of ovarian development. RNAi experiment and trilostane injection results showed that Sp HSD3B had regulatory effects on several genes related to gonadal development and steroid hormone synthesis. 15-day trilostane suppression could also inhibit ovarian development and progesterone level of hemolymph. According to the above results, crustaceans may have steroid hormone synthesis pathways like vertebrates, and the Hsd3b gene may be involved in the gonadal development of crabs. This study provides further insight into the function of genes involved in steroid hormone synthesis in crustaceans. • Steroid hormone synthesis pathway-related gene— Hsd3b was identified from Scylla paramamosain. • SpHsd3b knockdown can decrease the expression of gonadal development and steroid hormone synthesis-related genes. • Trilostane can inhibit ovarian development and lower progesterone level of hemolymph. • crustaceans may have steroid hormone synthesis pathways like vertebrate. [ABSTRACT FROM AUTHOR]

• Sp-IAG promoter has a dsx and a foxl-2 binding sites that can regulate its expression. • Cell experiment and RNAi experiment proved that dsx can enhance Sp-IAG expression. • Cell experiment and RNAi experiment showed that foxl-2 can inhibit Sp-IAG expression. Scylla paramamosain is an important cultured crab species on the southeast coast of China. However, the molecular regulation mechanism of its gonadal development still has not been thoroughly studied. Dsx (doublesex) and foxl-2 (forkhead transcription factor gene 2) are important transcription factors involved in gonadal development. So far, studies on the functions of dsx and foxl-2 in crustaceans are very limited. Insulin-like androgenic gland hormone (IAG) is an effector molecule that regulates the differentiation, development and sex maintenance of testes in crustaceans. In this study, the promoter region of Sp-IAG was predicted, and several potential binding sites of dsx and foxl-2 were found. Site-directed mutagenesis was performed on the predicted potential binding sites, and their promoter activity was analyzed. The results showed that there was a dsx and a foxl-2 binding site, respectively, that could regulate the expression of Sp-IAG. In order to verify the regulatory effect of these two transcription factors on Sp-IAG , we constructed the expression plasmids of dsx and foxl-2 and co-transfected them into HEK293T cell lines with the promoter of Sp-IAG , respectively. The results showed that dsx could significantly promote the expression of Sp-IAG , while foxl-2 could inhibit its expression substantially. Then we carried out in vivo RNA interference experiment on mud crabs. The expression of dsx and foxl-2 in crabs was interfered respectively. The results of qRT-PCR showed that the expression of Sp-IAG was significantly inhibited after interfering with dsx , while significantly increased after interfering with foxl-2 , which was consistent with the cell experiment. In conclusion, dsx and foxl-2 transcription factors play opposite roles in regulating the expression of Sp-IAG. [ABSTRACT FROM AUTHOR]

High seawater temperatures in summer often lead to high mortality in farmed abalone, which greatly increases the cost of farming. The hybrid abalone (Haliotis discus hannai ♀ × H. fulgens ♂, DF), is one of the main varieties of abalone cultured in Fujian Province, China, to resist the high temperature. In this study, the high-temperature resistance of DF abalones was assessed under 28 °C - 32 °C. 31 °C was applied in the subsequent screening experiments of high-temperature resistant individuals. A total of 614,321 single nucleotide polymorphisms (SNPs) were identified via super genotyping-by-sequencing (S-GBS), and 69 SNPs involved in 42 candidate genes were identified by genome-wide association study (GWAS). In addition, according to gene functional annotation, the locus with the lowest P value was screened within the threshold range (−log 10 P = 4.98296). The genotyping was verified by Sequenom MassARRAY, and 7 SNPs were proved to be related to high temperature resistance in DF population of 100 individuals. 7 candidate genes (fabp , e3rglg4 , rgs2 , tanc2x6 , osbp1 , arsb , and uch1) were identified near these SNPs. Finally, these candidate genes were further proved to be associated with the high-temperature resistance trait of DF by the analysis of differential expression genes between sensitive and tolerant individuals. These candidate genes involved regulatory mechanisms such as cell stability, protein repair, lipid metabolism and immune regulation. Notably, the mechanism by which the e3rglg4 gene was involved in the degradation of abnormally folded proteins of abalone under heat stress, corroborating the reliability of the results of GWAS and Sequenom MassARRA. These results complement the shortage of molecular markers for heat resistance of hybrid abalone, provide a reference for marker-assisted selection (MAS) and accelerate the development of heat-resistant abalone varieties. [Display omitted] • This is the first study to detect high-temperature resistance-related SNPs and genes in hybrid abalone. • A total of seven SNPs were found to be highly associated with high temperature resistance. • Seven genes were identified as involved in regulating DF (Haliotis discus hannai ♀ × H. fulgens ♂) high temperature resistance. [ABSTRACT FROM AUTHOR]

• The full-length cDNAs of alp and acp of Scylla paramamosain were identified. • Expression of Spalp and Spacp were the highest levels in the hepatopancreas. • The core promoter region of the 5′-flanking region of Spalp and Spacp were confirmed. • Potential transcription factor binding sites were found in the 5′-flanking regions. • This study is the first one to analyze the activity of Spalp and Spacp promoters. In crustaceans, innate immunity serves as the frontline of defense against microbes. Alkaline phosphatases (ALPs) and acid phosphatases (ACPs) are essential enzymes that play a significant role in crustaceans' immune defenses. However, the function and transcriptional regulation of the alp and acp genes in the Scylla paramamosain , an important aquaculture species in China, have not been elucidated. In this study, the full-length cDNAs of Spalp and Spacp were identified, which consist of 2,718 bp and 3,768 bp, encoding 579 and 452 amino acids, respectively. Multiple sequence alignment and phylogenetic analysis showed that these two genes were conserved among different species and shared high homology with crustaceans. The mRNA expression of Spalp and Spacp were examined in eight tested tissues, with the highest levels in the hepatopancreas. The 5′-flanking regions of Spalp and Spacp were cloned and sequenced. The core promoter region of the Spalp and Spacp was −39 bp∼+8 bp and −39 bp∼+10 bp, respectively. Potential binding sequences for SOX-2, c-fos, SP1, NF-κB, GATA-1, YY1, and AP-1 transcription factors were found in the 5′-flanking regions of Spalp and Spacp. The NF-κB binding site located between −1,223 bp and −972 bp in Spalp while SP1 and AP-1 binding sites located between −1,249 bp and −514 bp in Spacp. Mutation analysis confirmed that NF-κB negatively regulated the expression of Spalp gene, and SP1 and AP-1 positively regulated Spacp gene expression. These results provide us with essential information to elucidate the function of the Spalp and Spacp in S. paramamosain. This study is the first one to analyze the activity of Spalp and Spacp promoters. [ABSTRACT FROM AUTHOR]

Fish cell lines are important for the study in resource conservation, genetic breeding, diseases control, and environmental pollutants detection. The establishment of gonadal cell lines will provide a suitable in vitro model for studying the molecular regulation mechanisms underlying sex determination and sex-related gene expression and function. Large yellow croaker (Larimichthys crocea) is the unique marine economic fish with the highest production in China's coastal waters. In this study, the continuous gonadal cell lines, ovary cell line of large yellow croaker (LYCO) and testis cell line of large yellow croaker (LYCT), derived from the ovary and testis of juvenile fish were established with modified Leibovitz's L-15 medium at 27 °C. The medium was perfected to improve the growth rate and the increment ability of the LYCO and LYCT. The characteristics of the two cell lines were investigated, including the concentration of zeta-life fetal bovine serum (FBS), different passage ratios, cryopreservation, and recovery. The cell lines were successfully transfected with pEGFP-N1 and pNanog-N1 by electroporation, and the transfection efficiency of cells was high; these results showed that the cell lines could be used to study the expression of exogenous and endogenous genes. The LYCO expressed the marker gene of ovarian follicle cells, Foxl2 , while the LYCT expressed the marker gene of testicular sertoli cells, Dmrt1. However, LYCO and LYCT did not express Vasa , a marker gene of germ cells. Therefore, LYCO and LYCT appeared to consist of ovarian follicle cells and testicular sertoli cells, respectively. After the knockdown of Dmrt1 and Foxl2 in LYCO and LYCT, respectively, the results showed that the cell lines could be effective for RNA-mediated interference (RNAi) experiments and the study of the interactions among genes. As a first fish gonadal cell lines of L. crocea, which can provide a new model for the interactions between somatic cells and germ cells, and a basis for the further study of their reproduction and breeding in L.crocea. The results lay a foundation for the study of the gonadal cell culture in marine fish in the future. • Two cell lines from the ovary and testis of large yellow croaker (LYCO and LYCT) were first developed. • LYCO and LYCT can be used for high-efficiency and rapid electroporation to induce transformation to study gene function. • Overexpression of Nanog in LYCO and LYCT laid a foundation for the study of endogenous gene expression. • It is the first time to report the relationship of the sex-related genes by Dmrt1 and Foxl2 knockdown in LYCO and LYCT. [ABSTRACT FROM AUTHOR]

Scylla paramamosain is an economically important cultured crab species in China. Cyclins and cyclin-dependent kinases (CDKs) play important roles in regulations of cell cycle and ovarian development. MiRNAs can negatively regulate gene expression at the post-transcriptional level through base-complementary pairing with the 3'-untranslated region (3-UTR) of the target gene. In this study, bioinformatics prediction showed that miR-9c and miR-263a identified from our group's gonad miRNAome of S. paramamosain may bind to the 3' UTR region of cyclin A , cyclin B , cyclin E , cyclin H , CDK1, and CDK2. Furthermore, the results of double luciferase reporter gene assay showed that the luciferase activities of HEK293T cells co-transfected with miR-9c mimics/miR-9c inhibitor and the 3'-UTR plasmid vectors of the five genes (cyclin A , cyclin B , cyclin H , CDK1, and CDK2) were significantly decreased/increased compared with those in the NC (negative control) and BC (blank control) groups. The results in miR-263a were similar to miR-9c, but all of the six genes could be regulated by miR-263a. In in vivo experiments, agomiR-9c (miR-9c enhancer) injection resulted in decreases of cyclin A and CDK1 expression level, and reverse effects were observed by injecting antagomiR-9c. AgomiR-263a decreased the expression of cyclin A , cyclin B , cyclin H , CDK1, and CDK2, but antagomiR-263a increased their expression. Both the in vitro and in vivo experiments confirmed functions of miR-9c and miR-263a in cell cycle progress of ovarian development by expression regulation of cyclin A , cyclin B , cyclin E , cyclin H , CDK1, and CDK2. The findings provide new insights into the reproductive regulation mechanism in mud crab and further enrich the knowledge of cell cycle and ovarian development regulation in invertebrates. [Display omitted] • The regulation of miR-9c and miR-263a on cyclins and CDKs was confirmed in Scylla paramamosain. • MiR-263a can inhibit the expression of cyclin A , cyclin B , cyclin E , cyclin H , CDK1 , and CDK2. • MiR-9c only inhibit the expression of cyclin A and CDK1. • MiR-263a may play a more essential role than miR-9c in regulating cell cycle genes in S. paramamosain. • The change of vtg expression level manifests that cyclins and CDKs can regulate ovarian development. [ABSTRACT FROM AUTHOR]

Background: The mud crab Scylla paramamosain is an economically important species for aquaculture in China and has sexually dimorphic between females and males. Understanding sex differentiation in this species is essential for the development of monosex aquaculture. The Dmrt genes play a vital role in sex differentiation in animals., Methods and Results: In this study, two dmrt-like transcript variants, Spdmrt-like-tv1 and Spdmrt-like-v2, were cloned. SpDmrt-like-tv1 contained a DM domain, while SpDmrt-like-tv2 contained a DM and a DMA domain. Spdmrt-like-tv1 and Spdmrt-like-tv2 were both specifically expressed in testis. During testicular development, the expression level of Spdmrt-like-tv1 increased from stage I to stage II (P > 0.05) and then decreased from stage II to stage III (P < 0.05). The expression level of Spdmrt-like-tv2 in stages I and II was significantly higher than that in stage III (P < 0.05). During embryonic development, the expression level of Spdmrt-like-tv1 was higher in the mid-embryonic stage compared with the early and late stages, but the differences were not significant. Moreover, the expression level of Spdmrt-like-tv2 was stable and remained high throughout embryonic development. Furthermore, the expression level of Spdmrt-like-tv2 was significantly higher than that of Spdmrt-like-tv1. Knockdown of Spdmrt-like variants indicated that the regulative target gene of Spdmrt-like-tv1 was Spsox21, and the regulative target genes of Spdmrt-like-tv2 were Spfoxl2 and Spsox21. Combined with the results in our previously published peer-reviewed articles that the expression of Spfoxl2 in the testis was significantly higher than that in the ovary, and Spfoxl2 negatively regulated Spvtg expression. Spsox21 played a role in the development and maintenance of testis as well as in the process of neural development and regulation of body segmentation., Conclusion: Therefore, we suggest that Spdmrt-like-tv1 and Spdmrt-like-tv2 might be involved in testicular development and embryonic development, and Spdmrt-like-tv2 might play more important roles in these two developmental processes by regulating the expression of Spfoxl2 and Spsox21 due to its high expression., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

The red-spotted grouper Epinephelus akaara (E. akaara) is one of the most economically important marine fish in China, Japan and South-East Asia and is a threatened species. The species is also considered a good model for studies of sex inversion, development, genetic diversity and immunity. Despite its importance, molecular resources for E. akaara remain limited and no reference genome has been published to date. In this study, we constructed a chromosome-level reference genome of E. akaara by taking advantage of long-read single-molecule sequencing and de novo assembly by Oxford Nanopore Technology (ONT) and Hi-C. A red-spotted grouper genome of 1.135 Gb was assembled from a total of 106.29 Gb polished Nanopore sequence (GridION, ONT), equivalent to 96-fold genome coverage. The assembled genome represents 96.8% completeness (BUSCO) with a contig N50 length of 5.25 Mb and a longest contig of 25.75 Mb. The contigs were clustered and ordered onto 24 pseudochromosomes covering approximately 95.55% of the genome assembly with Hi-C data, with a scaffold N50 length of 46.03 Mb. The genome contained 43.02% repeat sequences and 5,480 noncoding RNAs. Furthermore, combined with several RNA-seq data sets, 23,808 (99.5%) genes were functionally annotated from a total of 23,923 predicted protein-coding sequences. The high-quality chromosome-level reference genome of E. akaara was assembled for the first time and will be a valuable resource for molecular breeding and functional genomics studies of red-spotted grouper in the future., (© 2019 John Wiley & Sons Ltd.)

Inhibitor of NF-κB (IκB), nuclear factor-κB (NF-κB), and Akirin2 are all important members of Rel/NF-κB signaling pathway, which plays a pivotal role in regulating the innate immune response of vertebrates and invertebrates. In this study, the IκB (SaIκB) and Akirin2 (SaAkirin2) cDNAs of small abalone Haliotis diversicolor were cloned and characterized. The full length cDNA of SaIκB and SaAkirin2 were 1748 bp and 1452 bp respectively, encoding a protein of 401 aa and 187 aa respectively. A conserved degradation motif (DS56GIYS60) and six ankyrin repeats were identified in the SaIκB by SMART analysis. Meanwhile, a typical nuclear localization signal (NLS) was found at the N-terminal region of the SaAkirin2 protein. Also, the mRNA expression level of SaIκB, SaAkirin2, and AbNF-κB were detected by quantitative real-time PCR. The results revealed that all these three genes were ubiquitously expressed in 7 selected tissues. The expression level of SaIκB in gills was higher than that in other tissues (P < 0.05) while the expression level of AbNF-κB was significantly higher in hepatopancreas and haemocytes. The highest expression level of SaAkirin2 was detected in hepatopancreas, followed by mantle. The mRNA expression levels in either gills or haemocytes of SaIκB, SaAkirin2, and AbNF-κB were significantly up-regulated (P < 0.05) post thermal stress, hypoxia exposure, thermal plus hypoxia stress and the injection of Vibrio parahaemolyticus. These results indicated that these three NF-κB signaling pathway-related genes are involved in response to bacterial infection and play essential roles in response to thermal and hypoxia stress., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The macrophage migration inhibitory factor (mif) cDNA and its genome were cloned from small abalone Haliotis diversicolor supertexta. Small abalone mif (samif) was originally identified from an expressed sequence tag (EST) fragment from a normalized cDNA library. It's 5' untranslated region (UTR) was obtained by 5' rapid amplification of cDNA end (RACE) techniques and its genomic DNA was cloned by PCR. The full-length cDNA of samif was of 535 bp, consisting of a 5'-terminal UTR of 49 bp, an open reading frame of 384 bp and a 3'-terminal UTR of 102 bp. The deduced protein was composed of 128 amino acids, with an estimated molecular mass of 14.0 kDa and a predicted pI of 6.90. The full-length samif genomic DNA comprises 3238 bp, containing three exons and two introns. Real time quantitative PCR analysis revealed that samif gene is constitutively expressed in 6 selected tissues, and its expression level in hepatopancreas is higher than that in the other tissues (p < 0.01). Samif expression level in the hepatopancreas at 24 and 48 h after Vibrio parahaemolyticus injection was upregulated significantly (p < 0.01), but there was no significant change after exposure to tributyltin (TBT) (p > 0.05).