Prevention and treatment of HPV-related cancer through a mRNA vaccine expressing APC-targeting antigen.

Persistent human papillomavirus (HPV) infection is associated with multiple malignancies. Developing therapeutic vaccines to eliminate HPV-infected and malignant cells holds significant value. In this study, we introduced a lipid nanoparticle encapsulated mRNA vaccine expressing tHA-mE7-mE6. Mutations were introduced into E6 and E7 of HPV to eliminate their tumourigenicity. A truncated influenza haemagglutinin protein (tHA), which binds to the CD209 receptor on the surface of dendritic cells (DCs), was fused with mE7-mE6 in order to allow efficient uptake of antigen by antigen presenting cells. The tHA-mE7-mE6 (mRNA) showed higher therapeutic efficacy than mE7-mE6 (mRNA) in an E6 and E7+ tumour model. The treatment resulted in complete tumour regression and prevented tumour formation. Strong CD8+ T-cell immune response was induced, contributing to preventing and curing of E6 and E7+ tumour. Antigen-specific CD8+ T were found in spleens, peripheral blood and in tumours. In addition, the tumour infiltration of DC and NK cells were increased post therapy. In conclusion, this study described a therapeutic mRNA vaccine inducing strong anti-tumour immunity in peripheral and in tumour microenvironment, holding promising potential to treat HPV-induced cancer and to prevent cancer recurrence.

Two-Color CRISPR Imaging Reveals Dynamics of Herpes Simplex Virus 1 Replication Compartments and Virus-Host Interactions.

Real-time imaging tools for single-virus tracking provide spatially resolved, quantitative measurements of viral replication and virus-host interactions. However, efficiently labeling both parental and progeny viruses in living host cells remains challenging. Here, we developed a novel strategy using the CRISPR-Tag system to detect herpes simplex virus 1 (HSV-1) DNA in host cells. We created recombinant HSV-1 harboring an ~600-bp CRISPR-Tag sequence which can be sufficiently recognized by dCas9-fluorescent protein (FP) fusion proteins. CRISPR-assisted single viral genome tracking (CASVIT) allows us to assess the temporal and spatial information of viral replication at the single-cell level. Combining the advantages of SunTag and tandem split green fluorescent protein (GFP) in amplifying fluorescent signals, dSaCas9-tdTomato10x and dSpCas9-GFP14x were constructed to enable efficient two-color CASVIT detection. Real-time two-color imaging indicates that replication compartments (RCs) frequently come into contact with each other but do not mix, suggesting that RC territory is highly stable. Last, two-color CASVIT enables simultaneous tracking of viral DNA and host chromatin, which reveals that a dramatic loss of telomeric and centromeric DNA occurs in host cells at the early stage of viral replication. Overall, our work has established a framework for developing CRISPR-Cas9-based imaging tools to study DNA viruses in living cells. IMPORTANCE Herpes simplex virus 1 (HSV-1), a representative of the family Herpesviridae, is a ubiquitous pathogen that can establish lifelong infections and widely affects human health. Viral infection is a dynamic process that involves many steps and interactions with various cellular structures, including host chromatin. A common viral replication strategy is to form RCs that concentrate factors required for viral replication. Efficient strategies for imaging the dynamics of viral genomes, RC formation, and the interaction between the virus and host offer the opportunity to dissect the steps of the infection process and determine the mechanism underlying each step. We have developed an efficient two-color imaging system based on CRISPR-Cas9 technology to detect HSV-1 genomes quantitatively in living cells. Our results shed light on novel aspects of RC dynamics and virus-host interactions.

A global analysis of CNVs in Chinese indigenous fine-wool sheep populations using whole-genome resequencing.

BACKGROUND: Copy number variation (CNV) is an important source of genetic variation that has a significant influence on phenotypic diversity, economically important traits and the evolution of livestock species. In this study, the genome-wide CNV distribution characteristics of 32 fine-wool sheep from three breeds were analyzed using resequencing. RESULTS: A total of 1,747,604 CNVs were detected in this study, and 7228 CNV regions (CNVR) were obtained after merging overlapping CNVs; these regions accounted for 2.17% of the sheep reference genome. The average length of the CNVRs was 4307.17 bp. "Deletion" events took place more frequently than "duplication" or "both" events. The CNVRs obtained overlapped with previously reported sheep CNVRs to variable extents (4.39-55.46%). Functional enrichment analysis showed that the CNVR-harboring genes were mainly involved in sensory perception systems, nutrient metabolism processes, and growth and development processes. Furthermore, 1855 of the CNVRs were associated with 166 quantitative trait loci (QTL), including milk QTLs, carcass QTLs, and health-related QTLs, among others. In addition, the 32 fine-wool sheep were divided into horned and polled groups to analyze for the selective sweep of CNVRs, and it was found that the relaxin family peptide receptor 2 (RXFP2) gene was strongly influenced by selection. CONCLUSIONS: In summary, we constructed a genomic CNV map for Chinese indigenous fine-wool sheep using resequencing, thereby providing a valuable genetic variation resource for sheep genome research, which will contribute to the study of complex traits in sheep.

Genome-wide association studies detects candidate genes for wool traits by re-sequencing in Chinese fine-wool sheep.

BACKGROUND: The quality and yield of wool determine the economic value of the fine-wool sheep. Therefore, discovering markers or genes relevant to wool traits is the cornerstone for the breeding of fine-wool sheep. In this study, we used the Illumina HiSeq X Ten platform to re-sequence 460 sheep belonging to four different fine-wool sheep breeds, namely, Alpine Merino sheep (AMS), Chinese Merino sheep (CMS), Aohan fine-wool sheep (AHS) and Qinghai fine-wool sheep (QHS). Eight wool traits, including fiber diameter (FD), fiber diameter coefficient of variance (FDCV), fiber diameter standard deviation (FDSD), staple length (SL), greasy fleece weight (GFW), clean wool rate (CWR), staple strength (SS) and staple elongation (SE) were examined. A genome-wide association study (GWAS) was performed to detect the candidate genes for the eight wool traits. RESULTS: A total of 8.222 Tb of raw data was generated, with an average of approximately 8.59X sequencing depth. After quality control, 12,561,225 SNPs were available for analysis. And a total of 57 genome-wide significant SNPs and 30 candidate genes were detected for the desired wool traits. Among them, 7 SNPs and 6 genes are related to wool fineness indicators (FD, FDCV and FDSD), 10 SNPs and 7 genes are related to staple length, 13 SNPs and 7 genes are related to wool production indicators (GFW and CWR), 27 SNPs and 10 genes associated with staple elongation. Among these candidate genes, UBE2E3 and RHPN2 associated with fiber diameter, were found to play an important role in keratinocyte differentiation and cell proliferation. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment results, revealed that multitude significant pathways are related to keratin and cell proliferation and differentiation, such as positive regulation of canonical Wnt signaling pathway (GO:0090263). CONCLUSION: This is the first GWAS on the wool traits by using re-sequencing data in Chinese fine-wool sheep. The newly detected significant SNPs in this study can be used in genome-selective breeding for the fine-wool sheep. And the new candidate genes would provide a good theoretical basis for the fine-wool sheep breeding.

Identification and functional characterizations of serpin8, a potential prophenoloxidase-activating protease inhibitor in Pacific white shrimp, Litopenaeus vannamei.

Serpins have been characterized from varieties of organisms by their inhibitory roles on serine or cysteine proteases. However, research for the functional study of serpins in crustacean is relatively small. To fully clarify the immune characterizations of serpin, a novel serpin (named Lvserpin8) encoding 414 amino acids with a 19-amino acid signal peptide and a serpin domain was identified from the Pacific white shrimp Litopenaeus vannamei. Sequence analysis indicated that the genomic Lvserpin8 gene contains 5 exons and 4 introns. The P1 residues of the predicted scissile bond in the reactive center loop (RCL) region represented for Lysine (Lys), which is in accordance with Pmserpin8, Dmserpin27A, Ofserpin3, Bmserpin3 and Msserpin3. Quantitative results showed that high mRNA expression of Lvserpin8 was detected in hepatopancreas and testis. Notably, a significant increase of Lvserpin8 was appeared post injection of Vibrio anguillarum, and Micrococcus lysodeikticus. Moreover, Lvserpin8 was knocked down in vivo by double-stranded RNA (dsRNA) mediated RNA interference (RNAi). Suppression of Lvserpin8 led to a significant increase in the transcripts of LvPPAE2 (Prophenoloxidase-activating Enzyme 2) and cumulative mortality. What's more, recombinant Lvserpin8 protein (rLvserpin8) displayed inhibition roles on trypsin activity, and prophenoloxidase activation. Taken together, the results implied that Lvserpin8 may be involved in shrimp innate immunity via the inhibition of prophenoloxidase-activating proteases.

Identification and characterization of two Croquemort homologues in penaeid shrimp Litopenaeus vannamei.

Croquemort, the homologue of human CD36, is a member of class B scavenger receptors, which is involved in bacteria phagocytosis and cytokins release. However, there is still less information about Croquemort in crustaceans. Here, a Croquemort from Pacific white shrimp Litopenaeus vannamei (LvCroquemort) and its truncated form (LvCroquemort-S1) cDNA sequences were identified, characterized and their role in bacteria clearance was investigated. The deduced protein of LvCroquemort is 533 amino acids and contains typical domains of CD36: the N-terminus and C-terminus in cytoplasm, two transmembrane regions and a large extracellular loop-like domain. However, LvCroquemort-S1 losses partial cDNA sequence in its middle and its deduced protein losses the C-terminal transmembrane region and C-terminus in cytoplasm, the latter of which is found participating in cytokins release in human CD36. LvCroquemort transcript is highly expressed in gills, hemocytes, testis and slightly in heart, hepatopancreas and nerve. Besides, its responses to bacteria Vibrio anguillarum and white spot syndrome virus were examined. Knock-down of LvCroquemort by specific dsRNA reduces bacteria clearance. These initial data will help to further understand roles of Croquemort in crustacean innate immunity.

Lvserpin3 is involved in shrimp innate immunity via the inhibition of bacterial proteases and proteases involved in prophenoloxidase system.

Serine protease inhibitor, represented by serpin, plays an important inhibitory role on proteases involved in the immune responses. To clarify the immune characterizations of serpin, a novel serpin (Lvserpin3) encoding for 410 amino acids with a 23-amino acid signal peptide and a serpin domain was identified from the Pacific white shrimp Litopenaeus vannamei. Lvserpin3 expressed strongest in hepatopancreas, and was significantly up-regulated in the early stage upon Vibrio anguillarum, Micrococcus lysodeikticus or White Spot Syndrome Virus (WSSV) infection. Suppression of Lvserpin3 by dsRNA led to a significant increase in the transcripts of LvPPAF, LvproPO and phenoloxidase (PO) activity, and also led to the high cumulative mortality. The recombinant Lvserpin3 protein (rLvserpin3) inhibited the proteases secreted by M. lysodeikticus and Bacillus subtilis, and further exhibited inhibitory role on the growth of B. subtilis and M. lysodeikticu. Moreover, rLvserpin3 was found to be able to block the activation of prophenoloxidase system. Taken together, the results imply that Lvserpin3 may be involved in shrimp innate immunity via the inhibition of bacterial proteases and proteases involved in prophenoloxidase system.

Identification of 10 transcripts of FREP in penaeid shrimp Litopenaeus vannamei.

Fibrinogen-related proteins (FREPs) are widely distributed in vertebrates and invertebrates and known having Fibrinogen-related domains (FReDs) which function in multiple aspects, especially in innate immune response as pattern recognition receptors. However, there is no any report about FREP in penaeid shrimp Litopenaeus vannamei. Here, totally 10 transcripts of FREP were isolated and named LvFREP1.1, 1.2 until 1.10. All of the 10 transcripts have high identity in their 3' ends and encode conserved FReDs. Since the 10 transcripts are highly similar we could not design any primers that can amplify a single transcript. We chose a pair of primers corresponding to part of LvFREP1.1 and LvFREP1.5 to examine their expression. Tissue distribution indicated LvFREP1.1 and LvFREP1.5 mRNA locates in hemocytes, gills, intestine, hearts and slightly in nerve. The expression of LvFREP1.1 and LvFREP1.5 behaves differently post bacteria and virus infection. Besides, recombinant LvFReD could agglutinate bacteria Vibrio harveyi in the presence of Ca2+. These initial data presents the diversity of FREPs in penaeid shrimp and also push us to further explore their roles in shrimp immune response.

Recombinant expression and characterization of a serine protease inhibitor (Lvserpin7) from the Pacific white shrimp, Litopenaeus vannamei.

Serine protease inhibitors (serpins) are widely known to its inhibitory role on proteases involved in the immune responses. Herein, a novel serine protease inhibitor (Lvserpin7), encoding for 411 amino acids with calculated molecular mass of 46.29 kDa and isoelectric point of 6.98 was characterized from the Pacific white shrimp Litopenaeus vannamei. Lvserpin7 shared 92.9% identities to Penaeus monodon serpin7. Among the tested tissues, Lvserpin7 was mainly expressed in hemocytes and gill. The expression profiles analysis indicated that Lvserpin7 was significantly up-regulated in the early stage upon Vibrio anguillarum, Micrococcus lysodeikticus or White Spot Syndrome Virus (WSSV) infection. Fusion protein expression was induced by IPTG, and the purified recombinant Lvserpin7 protein (rLvserpin7) binds to both the Gram-positive and Gram-negative bacteria. Also rLvserpin7 exhibited inhibitory activity against the proteases secreted by Bacillus subtilis. Moreover, rLvserpin7 showed inhibition role on prophenoloxidase activation. To recap, we proposed that Lvserpin7 was implicated in the shrimp immunity via the inhibition of bacterial proteases and proteases involved in prophenoloxidase system.

A galectin from shrimp Litopenaeus vannamei is involved in immune recognition and bacteria phagocytosis.

Galectins are conserved family members with ß-galactosides affinity that play multiple functions in embryogenesis, development and regulation of innate and adaptive immunity. However, little functional studies were reported in crustaceans. Here, a shrimp Litopenaeus vannamei galectin (LvGal) cDNA was identified with an open reading frame of 1017 bp, which encodes a putative protein of 338 amino acids. A carbohydrate recognition domain (CRD) and several amino acids residues involved in dimerization were found in LvGal. LvGal mRNA was mainly expressed in gills and hemocytes and upregulated post Vibrio anguillarum challenge. Recombinant LvGal (rLvGal) was expressed in Escherichia coli BL21 (DE3) and the purified rLvGal could strongly bind G(-) bacteria V. anguillarum and G(+) bacteria Micrococcus lysodeikticus. Besides, rLvGal exhibited strong activity to agglutinate V. anguillarum and weak activity to agglutinate M. lysodeikticus but no obvious antibacterial activity was found with selected bacteria. In addition, in vivo experiments showed rLvGal could promote phagocytosis of bacteria by hemocytes. Thus, through these collective data we predicted LvGal is involved in immune recognition and functions as a potential pattern recognition receptor.

Molecular characterization, immune responsive expression and functional analysis of QM, a putative tumor suppressor gene from the Pacific white shrimp, Litopenaeus vannamei.

The QM, firstly identified as a putative tumor suppressor gene from human, has been confirmed to possess varieties of functions in a range of organisms. In the present study, the cDNA that encodes a 220-amino-acid QM protein with calculated molecular mass of 25.5 kDa and isoelectric point of 10.07 was characterized from the Pacific white shrimp Litopenaeus vannamei. Analysis of the deduced amino acid sequence of LvQM revealed that it contained a series of conserved functional motifs. Quantitative real-time PCR (qRT-PCR) results showed that the transcript of LvQM was extensively distributed in the tissues under investigation and most highly expressed in gill. After challenged with Vibrio anguillarum, the LvQM transcripts were significantly increased (P < 0.05) both in hepatopancreas and hemocytes in the early experimental phase. When LvQM was knocked down by RNA interference (RNAi), the transcript of prophenoloxidase (proPO) and the phenoloxidase activity (PO) in shrimp hemolymph were dramatically decreased, while the mortality was significantly increased. Furthermore, the recombinant LvQM protein (rLvQM) was successfully expressed in Escherichia coli BL21 (DE3)-pLysS. Injecting the purified rLvQM mixed with V. anguillarum markedly increased the clearance rate of bacteria and PO activity in the shrimp hemolymph. Hence, we conclude that LvQM was involved in the host defense of L. vannamei, probably as a positive regulator to phenoloxidase activity.

Identification of ecdysteroid signaling late-response genes from different tissues of the Pacific white shrimp, Litopenaeus vannamei.

Ecdysteroids initiate signaling along multiple pathways that regulate various aspects of development, maturation, and reproduction in arthropods. This study was carried out to seek the late target genes of ecdysteroid signaling from different tissues of the Pacific white shrimp, Litopenaeus vannamei. In the present study, eight isoforms of ecdysteroid receptor (EcR), two isoforms of retinoic acid X receptor (RXR), and one homolog of E75 were characterized from L. vannamei. The overall protein sequences and specific functional sites of EcR, RXR and E75 among crustacean species were found highly conserved. Tissue-specific, development stage-specific, and molt stage-specific expression patterns of LvEcR, LvRXR, and LvE75 were detected by qPCR. Double stranded RNA (dsRNA)-mediated RNA interference (RNAi) of any one of the three genes LvEcR, LvRXR and LvE75 caused specific expression changes of the other two, and resulted in corresponding expression changes of two molting related genes Cathepsin-L (LvCHSL) and Hemocyanin (LvHCyn) in the hepatopancreas, two chitin metabolism related genes chitin synthase (LvChS) and chitinase isoenzyme (LvChi2) in the epidermis, and two muscle growth related genes LvActin and myosin heavy chain (LvMHC) in the muscle. In correspondence, after in vivo injections of 20 hydroxyecdysone, specific expression changes of LvEcR, LvRXR, LvE75, LvCHSL and LvHCyn in the hepatopancreas, LvEcR, LvRXR, LvE75, LvChS and LvChi2 in the epidermis, and LvEcR, LvRXR, LvE75, LvActin and LvMHC in the muscle were also observed, respectively. Results in our study indicate multiple functions of ecdysteroids signaling in L. vannamei and the function may be time- and space-specific; ecdysteroids may act through different pathways via its functional receptor heterodimer EcR-RXR and the early responsive gene E75 to perform specific regulation roles on the target genes in different shrimp tissues; LvCHSL and LvHCyn in the hepatopancreas, LvChS and LvChi2 in the epidermis, and LvActin and LvMHC in the muscle are potential targets for ecdysteroid control. This is the first report on nuclear receptors in the economically important shrimp L. vannamei.

cDNA cloning and expression analysis of myostatin/GDF11 in shrimp, Litopenaeus vannamei.

Myostatin (MSTN) and growth differentiation factor-11 (GDF11) are closely related proteins belonging to the transforming growth factor-ß (TGF-ß) superfamily. In vertebrates, MSTN is known to negatively regulate skeletal muscle growth, and GDF11 is found to inhibit neurogenesis. In invertebrates, only one ortholog of vertebrate MSTN and GDF11 (MSTN/GDF11) existed. Little attention has been paid on its role to date. In this study, the cDNA that encodes a 422-amino-acid MSTN/GDF11 protein (LvMSTN/GDF11) was characterized from a crustacean species, the Pacific white shrimp (Litopenaeus vannamei). Sequence analysis revealed that the overall protein sequence and specific functional sites of LvMSTN/GDF11 were highly conserved with those in other crustacean species. Expression analysis by quantitative real-time reverse transcription polymerase chain reaction technique demonstrated its tissue-specific, larval developmental stage-specific, and molt stage-specific expression pattern, respectively. After in vivo injections of 20 hydroxyecdysone (20E), LvMSTN/GDF11 transcripts were declined in the abdominal (A) and pleopod (P1) muscles, increased in the pereiopod (P2) muscle, and not affected in the thoracic (T) muscle. The observed expression profiles suggest multiple functions of LvMSTN/GDF11 in L. vannamei and its role differs during the larval development and natural molt cycle. The different responses of LvMSTN/GDF11 to acute increases of 20E in the A, P1, P2 and T muscles may indicate that LvMSTN/GDF11 is transcriptionally regulated via ecdysteroids to coincide with its specific roles in the former three muscles, while its role may be independent of 20E regulation in the T muscle.

mRNA vaccines encoding fusion proteins of monkeypox virus antigens protect mice from vaccinia virus challenge.

The recent outbreaks of mpox have raised concerns over the need for effective vaccines. However, the current approved vaccines have either been associated with safety concerns or are in limited supply. mRNA vaccines, which have shown high efficacy and safety against SARS-CoV-2 infection, are a promising alternative. In this study, three mRNA vaccines are developed that encode monkeypox virus (MPXV) proteins A35R and M1R, including A35R extracellular domain -M1R fusions (VGPox 1 and VGPox 2) and a mixture of encapsulated full-length mRNAs for A35R and M1R (VGPox 3). All three vaccines induce early anti-A35R antibodies in female Balb/c mice, but only VGPox 1 and 2 generate detectable levels of anti-M1R antibodies at day 7 after vaccination. However, all three mRNA vaccine groups completely protect mice from a lethal dose of vaccinia virus (VACV) challenge. A single dose of VGPox 1, 2, and 3 provide protection against the lethal viral challenge within 7 days post-vaccination. Long-term immunity and protection were also observed in all three candidates. Additionally, VGPox 2 provided better passive protection. These results suggest that the VGPox series vaccines enhance immunogenicity and can be a viable alternative to current whole-virus vaccines to defend against mpox.

Heterologous boost with mRNA vaccines against SARS-CoV-2 Delta/Omicron variants following an inactivated whole-virus vaccine.

The coronavirus SARS-CoV-2 has mutated quickly and caused significant global damage. This study characterizes two mRNA vaccines ZSVG-02 (Delta) and ZSVG-02-O (Omicron BA.1), and associating heterologous prime-boost strategy following the prime of a most widely administrated inactivated whole-virus vaccine (BBIBP-CorV). The ZSVG-02-O induces neutralizing antibodies that effectively cross-react with Omicron subvariants. In naïve animals, ZSVG-02 or ZSVG-02-O induce humoral responses skewed to the vaccine's targeting strains, but cellular immune responses cross-react to all variants of concern (VOCs) tested. Following heterologous prime-boost regimes, animals present comparable neutralizing antibody levels and superior protection against Delta and Omicron BA.1variants. Single-boost only generated ancestral and omicron dual-responsive antibodies, probably by "recall" and "reshape" the prime immunity. New Omicron-specific antibody populations, however, appeared only following the second boost with ZSVG-02-O. Overall, our results support a heterologous boost with ZSVG-02-O, providing the best protection against current VOCs in inactivated virus vaccine-primed populations.

Abundance of natural resources and environmental sustainability: the roles of manufacturing value-added, urbanization, and permanent cropland.

Sustainable management of natural resources and green urbanization is crucial because it assists the use of resources wisely without unnecessary use and without affecting future generations' needs. This research aims to examine the impact of the abundance of natural resources on China's CO2 emissions while moderating the roles of manufacturing value-added, urbanization, and permanent cropland from 1970 to 2016. This study developed a comprehensive empirical analysis, applied advanced econometric methodologies, and used the generalized linear model (GLM) and robust generalized estimating equation (GEE). Overall, the results conclude that natural resource abundance and permanent cropland are negatively associated with China's CO2 emissions. However, urbanization and manufacturing value-added are negatively related to those CO2 emissions. Moreover, natural resource abundance and permanent cropland improve environmental sustainability while urbanization and manufacturing value-added deteriorate that environmental sustainability. It is suggested that policymakers should promote sustainable management of natural resources and encourage economic usage of natural resources to boost resilient ecosystems; shape sustainable places, lifestyles, and communities; and consume natural resources less. Additionally, policymakers should consider collaborating with landscape architects, urban planners, engineers, transport planners, ecologists, sociologists, physiologists, economists, physicists, and other specialists to develop green urban communities. The limitations of the study and directions for future research are discussed.

Clean energy investment and financial development as determinants of environment and sustainable economic growth: evidence from China.

Environmental sustainability has become one of the most common phrases in discussions about climate change. This study examines the impact of clean energy investment and financial development on environmental sustainability and China's economic growth, using manufacturing value-added and urbanization as moderator variables from 1970 to 2016. We used advanced econometric methodologies for empirical estimations, used structural break unit root tests, fully modified least square, dynamic least square, and robust least square multiple regressions for long-run estimates. Overall, the results determine that clean energy investment is negatively associated with CO2 emissions and ecological footprint while positively associated with China's economic growth. Financial development, manufacturing value-added, and urbanization are positively associated with CO2 emissions, ecological footprint, and China's economic growth. Moreover, clean energy investment improves environmental sustainability at the expense of economic growth. Financial development, manufacturing value-added, and urbanization encourage economic growth at the expense of environmental sustainability. We argued that the local governments play a critical role in lifting the outstanding barriers to cleaner energy investment, addressing disincentives, including pricing carbon dioxide emissions, reforming inefficient nonrenewable fossil fuel subsidies, and addressing regulatory and market rigidities that can undesirably affect the attractiveness of clean energy investment. Policymakers are suggested to encourage green finance strategy for the financial sector to broader sustainable development objectives. At the heart of green manufacturing, industrialization policies are needed to integrate diverse intentions, like inclusive growth, environmental protection, and productivity through a wider range of economic, social, and environmental policy frameworks suitable for decoupling growth from social and environmental unsustainability.

Complex q-rung orthopair fuzzy Frank aggregation operators and their application to multi-attribute decision making.

The complex q-rung orthopair fuzzy sets (Cq-ROFSs) can serve as a generalization of q-rung orthopair fuzzy sets (q-ROFSs) and complex fuzzy sets FS (CFSs). Cq-ROFSs provide more freedom for people handling uncertainty and vagueness by the truth and falsity grades on the condition that the sum of the q-powers of the real part and imaginary part is within the unit interval. Further, Frank operational laws are an extended form of Archimedes' T mode and Archimedes' S mode and Frank aggregation operators have a certain parameter which makes them more flexible and more generalized than many other aggregation operators in the process of information fusion. The objectives of this paper are to extend the Frank operations to the complex q-rung orthopair fuzzy environment and to introduce their score function and accuracy function. Meanwhile, some complex q-rung fuzzy Frank aggregation operators are developed, such as the complex q-rung orthopair fuzzy Frank weighted averaging (Cq-ROFFWA) operator, the complex q-rung orthopair fuzzy Frank weighted geometric (Cq-ROFFWG) operator, and the complex q-rung orthopair fuzzy Frank ordered weighted averaging (Cq-ROFFOWA) operator, and their special cases are discussed. In addition, an innovative MADM method is introduced according to the propounded operators to deal with multi-attribute decision-making problems under the complex q-rung orthopair fuzzy environment. Consequently, the practicability and effectiveness of the created methods are proposed by parameter exploration and comparative analysis.

Decomposition and decoupling research of Chinese power sector carbon emissions through the consumption accounting principle.

The Logarithmic Mean Divisia Index (LMDI) model is applied to study Chinese national and regional power sector carbon emission changes through consumption side from 2003 to 2017, and regional power sector carbon emissions are estimated through the production and consumption accounting principle. The two-factor ANOVA and one-factor ANOVA are used to compare the differences of regional power sector carbon emissions through the two principles. In addition, the Tapio decoupling analysis model is used to investigate the decoupling state between carbon emissions of power sector and the corresponding driving forces through the consumption side. There are several results: (1) Through the two different principles, regional power sector carbon emissions are statistically significant, yet national power sector carbon emissions are not statistically significant; (2) the main factors contributing to the power sector carbon emission growth are economic scale effect and income level effect, and the main restraining factors are electricity consumption carbon intensity effect and production sector electricity intensity effect; (3) the highest contribution effect to the decoupling indexes between various influencing factors and power sector carbon emissions was scale effect, and technical effect had the second largest contribution value; (4) in 2003-2017, economic scale effect was the first significant factor causing the difference of regional power sector carbon emissions, followed by production sector electricity intensity effect and electricity consumption carbon intensity through the regional decomposition analysis. Finally, this paper gives some targeted suggestions for the low-carbon development of the power sector through national and regional perspectives.

Interactome and Ubiquitinome Analyses Identify Functional Targets of Herpes Simplex Virus 1 Infected Cell Protein 0.

Herpes simplex virus 1 (HSV-1) can productively infect multiple cell types and establish latent infection in neurons. Infected cell protein 0 (ICP0) is an HSV-1 E3 ubiquitin ligase crucial for productive infection and reactivation from latency. However, our knowledge about its targets especially in neuronal cells is limited. We confirmed that, like in non-neuronal cells, ICP0-null virus exhibited major replication defects in primary mouse neurons and Neuro-2a cells. We identified many ICP0-interacting proteins in Neuro-2a cells, 293T cells, and human foreskin fibroblasts by mass spectrometry-based interactome analysis. Co-immunoprecipitation assays validated ICP0 interactions with acyl-coenzyme A thioesterase 8 (ACOT8), complement C1q binding protein (C1QBP), ovarian tumour domain-containing protein 4 (OTUD4), sorting nexin 9 (SNX9), and vimentin (VIM) in both Neuro-2a and 293T cells. Overexpression and knockdown experiments showed that SNX9 restricted replication of an ICP0-null but not wild-type virus in Neuro-2a cells. Ubiquitinome analysis by immunoprecipitating the trypsin-digested ubiquitin reminant followed by mass spectrometry identified numerous candidate ubiquitination substrates of ICP0 in infected Neuro-2a cells, among which OTUD4 and VIM were novel substrates confirmed to be ubiquitinated by transfected ICP0 in Neuro-2a cells despite no evidence of their degradation by ICP0. Expression of OTUD4 was induced independently of ICP0 during HSV-1 infection. Overexpressed OTUD4 enhanced type I interferon expression during infection with the ICP0-null but not wild-type virus. In summary, by combining two proteomic approaches followed by confirmatory and functional experiments, we identified and validated multiple novel targets of ICP0 and revealed potential restrictive activities of SNX9 and OTUD4 in neuronal cells.