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1.
Ophthalmic Plast Reconstr Surg ; 40(3): 346-351, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38738712

RESUMO

PURPOSE: Scar contracture of the eyelid following facial burns often has adverse consequences. Total cicatricial contracture often makes adjustment flap translation challenging to implement. Previously used upper and lower eyelid adhesion methods are ineffective for patients with severe cicatricial contracture, and ectropion can easily recur. This study aimed to retrospectively examine upper and lower eyelid adhesions using an orbicularis oculi muscle flap and verify its stability. METHODS: In patients with ectropion caused by severe scar contracture following head and face burns, we employed a tunnel orbicularis oculi muscle flap technique, which involved creating a tunnel between the skin and the tarsal plate of the eyelid, mobilizing the orbicularis oculi muscle, and rotating it into this tunnel to provide stable adhesion of the upper and lower eyelids. Full-thickness skin grafting was then performed. The eyelids were examined postoperatively to determine whether reoperation was necessary and to monitor for any potential complications. RESULTS: This study included 26 patients and 46 eyes. No accidental disconnection occurred after eyelid adhesion, which lasted for an average of 21.87 ± 10.08 months before the eyelid adhesion was cut open. No complications or adverse reactions occurred, and the adhesions did not break unexpectedly. CONCLUSIONS: Repairing eyelid ectropion with the tunnel orbicularis oculi muscle flap is a simple procedure that immediately creates tension against upper and lower eyelid contractures, providing long-term stable adhesion. This method avoids structural disorders, such as eyelid margin scarring, minimally influences surrounding tissues, and has few postoperative complications. It holds great value for repairing eyelid tissue defects and warrants further study.


Assuntos
Ectrópio , Pálpebras , Músculos Oculomotores , Transplante de Pele , Retalhos Cirúrgicos , Humanos , Ectrópio/cirurgia , Ectrópio/etiologia , Masculino , Feminino , Estudos Retrospectivos , Adulto , Transplante de Pele/métodos , Pessoa de Meia-Idade , Músculos Oculomotores/cirurgia , Músculos Oculomotores/fisiopatologia , Adulto Jovem , Pálpebras/cirurgia , Blefaroplastia/métodos , Adolescente , Criança , Aderências Teciduais/cirurgia , Aderências Teciduais/etiologia , Queimaduras Oculares/cirurgia , Queimaduras Oculares/fisiopatologia , Queimaduras Oculares/diagnóstico
2.
Mol Ecol ; 32(14): 3859-3871, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37194687

RESUMO

Domesticated honeybees and wild bees are some of the most important beneficial insects for human and environmental health, but infectious diseases pose a serious risk to these pollinators, particularly following the emergence of the ectoparasitic mite Varroa destructor as a viral vector. The acquisition of this novel viral vector from the Asian honeybee Apis ceranae has fundamentally changed viral epidemiology in its new host, the western honeybee A. mellifera. While the recently discovered Lake Sinai Viruses (LSV) have been associated with weak honeybee colonies, they have not been associated with vector-borne transmission. By combining a large-scale multi-year survey of LSV in Chinese A. mellifera and A. cerana honeybee colonies with globally available LSV-sequence data, we investigate the global epidemiology of this virus. We find that globally distributed LSV is a highly diverse multi-strain virus, which is predominantly associated with the western honeybee A. mellifera. In contrast to the vector-borne deformed wing virus, LSV is not an emerging disease. Instead, demographic reconstruction and strong global and local population structure indicates that it is a highly variable multi-strain virus in a stable association with its main host, the western honeybee. Prevalence patterns in China suggest a potential role for migratory beekeeping in the spread of this pathogen, demonstrating the potential for disease transmission with the man-made transport of beneficial insects.


Assuntos
Abelhas , Vírus de RNA , Varroidae , Animais , Humanos , Abelhas/parasitologia , Abelhas/virologia , China/epidemiologia , Vírus de RNA/genética , Varroidae/virologia , Vírus
3.
J Gen Virol ; 103(8)2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35947094

RESUMO

Black queen cell virus (BQCV) is a severe threat to the honeybee (Apis mellifera) worldwide. Although several BQCV strains have been reported in China, the molecular basis for BQCV pathogenicity has not been well understood. Thus, a reverse genetic system of BQCV is required for studying viral replication and its pathogenic mechanism. Here, the complete genome sequence of BQCV was obtained from honeybees using reverse transcription PCR (RT-PCR), namely a BQCV China-GS1 strain (KY741959). Then, a phylogenetic tree was built to analyse the genetic relationships among BQCV strains from different regions. Our results showed that the BQCV China-GS1 contained two ORFs, consistent with the known reference strains, except for the BQCV China-JL1 strain (KP119603). Furthermore, the infectious clone of BQCV was constructed based on BQCV China-GS1 using a low copy vector pACYC177 and gene recombination. Due to the lack of culture cells for bee viruses, we infected the healthy bees with infectious clone of BQCV, and the rescued BQCV resulted in the recovery of recombinant virus, which induced higher mortality than those of the control group. Immune response after inoculated with BQCV further confirmed that the infectious clone of BQCV caused the cellular and humoral immune response of honeybee (A. mellifera). In conclusion, the full nucleotide sequence of BQCV China-GS1 strain was determined, and the infectious clone of BQCV was constructed in this study. These data will improve the understanding of pathogenesis and the host immune responses to viral infection.


Assuntos
Dicistroviridae , Vírus de RNA , Vírus , Animais , Abelhas , Dicistroviridae/genética , Fases de Leitura Aberta , Filogenia , Vírus de RNA/genética , Vírus/genética
4.
Virol J ; 18(1): 83, 2021 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-33882983

RESUMO

BACKGROUND: Sacbrood is an infectious disease of the honey bee caused by Scbrood virus (SBV) which belongs to the family Iflaviridae and is especially lethal for Asian honeybee Apis cerana. Chinese Sacbrood virus (CSBV) is a geographic strain of SBV. Currently, there is a lack of an effective antiviral agent for controlling CSBV infection in honey bees. METHODS: Here, we explored the antiviral effect of a Chinese medicinal herb Radix isatidis on CSBV infection in A. cerana by inoculating the 3rd instar larvae with purified CSBV and treating the infected bee larvae with R. isatidis extract at the same time. The growth, development, and survival of larvae between the control and treatment groups were compared. The CSBV copy number at the 4th instar, 5th instar, and 6th instar larvae was measured by the absolute quantification PCR method. RESULTS: Bioassays revealed that R. isatidis extract significantly inhibited the replication of CSBV, mitigated the impacts of CSBV on larval growth and development, reduced the mortality of CSBV-infected A. cerana larvae, and modulated the expression of immune transcripts in infected bees. CONCLUSION: Although the mechanism underlying the inhibition of CSBV replication by the medicine plant will require further investigation, this study demonstrated the antiviral activity of R. isatidis extract and provides a potential strategy for controlling SBV infection in honey bees.


Assuntos
Antivirais , Abelhas/virologia , Extratos Vegetais , Plantas Medicinais , Vírus de RNA/efeitos dos fármacos , Animais , Antivirais/farmacologia , Larva , Extratos Vegetais/farmacologia , Plantas Medicinais/química
5.
Environ Sci Technol ; 55(17): 11680-11692, 2021 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-34374532

RESUMO

Microplastics (MPs) are an emerging threat to ecological conservation and biodiversity; however, little is known of the types and possible impacts of MPs in pollinators. To examine whether MPs were present in honeybees, we analyzed the honeybee samples collected in fields from six provinces in China. Four types MPs were identified in honeybee including polystyrene (PS) by Raman spectroscopic analysis, and these plastic polymers were detected in 66.7% bee samples. Then, we assessed the physical and biological impacts of PS of three sizes (0.5, 5, and 50 µm) on bees for 21 days. Next, we measured how the presence of PS affected the Israeli acute paralysis virus proliferation, a small RNA virus associated with bee colony decline. Experimental evidence showed that a large mass of PS was ingested and accumulated within the midgut and enhanced the susceptibility of bees to viral infection. Not only histological analysis showed that PS, especially 0.5 µm PS, damaged the midgut tissue and was subsequently transferred to the hemolymph, trachea, and Malpighian tubules, but also qPCR and transcriptomic results indicated that genes correlated with membrane lipid metabolism, immune response, detoxification, and the respiratory system were significantly regulated after PS ingestion. Our results highlight neglected MP contamination to the bees, a pollination ecosystem stressed by the anthropogenic pollution, and have implications for human health via ingestion of bee products.


Assuntos
Microplásticos , Viroses , Animais , Abelhas , Ingestão de Alimentos , Ecossistema , Plásticos , Poliestirenos
6.
Int J Mol Sci ; 22(18)2021 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-34576241

RESUMO

Although it had been reported that Israeli acute paralysis virus (IAPV) can cause systemic infection in honey bees, little is known about how it establishes this infection and results in the typical symptoms, paralysis and trembling. Here, we used our previously constructed IAPV infectious clone to investigate viral loads in different tissues of honey bees and further identify the relation between tissue tropism and paralytic symptoms. Our results showed that tracheae showed a greater concentration of viral abundance than other tissues. The abundance of viral protein in the tracheae was positively associated with viral titers, and was further confirmed by immunological and ultrastructural evidence. Furthermore, higher viral loads in tracheae induced remarkable down-regulation of succinate dehydrogenase and cytochrome c oxidase genes, and progressed to causing respiratory failure of honey bees, resulting in the appearance of typical symptoms, paralysis and body trembling. Our results showed that paralysis symptoms or trembling was actually to mitigate tachypnea induced by IAPV infection due to the impairment of honey bee tracheae, and revealed a direct causal link between paralysis symptoms and tissue tropism. These findings provide new insights into the understanding of the underlying mechanism of paralysis symptoms of honey bees after viral infection and have implications for viral disease prevention and specific therapeutics in practice.


Assuntos
Dicistroviridae , Paralisia/fisiopatologia , Taquipneia/fisiopatologia , Viroses/fisiopatologia , Animais , Abelhas/virologia , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Paralisia/virologia , Succinato Desidrogenase/metabolismo , Taquipneia/virologia , Traqueia/virologia , Carga Viral , Proteínas Virais , Viroses/virologia
7.
Appl Microbiol Biotechnol ; 104(1): 257-276, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31754765

RESUMO

Currently, knowledge of circular RNAs (circRNAs) in insects including honeybee is extremely limited. Here, differential expression profiles and regulatory networks of circRNAs in the midguts of Apis cerana cerana workers were comprehensively investigated using transcriptome sequencing and bioinformatics. In total, 9589 circRNAs (201-800 nt in length) were identified from 8-day-old and 11-day-old workers' midguts (Ac1 and Ac2); among them, 5916 (61.70%) A. cerana cerana circRNAs showed conservation with our previously indentified circRNAs in Apis mellifera ligucstica workers' midguts (Xiong et al., Acta Entomologica Sinica 61:1363-1375, 2018). Five circRNAs were confirmed by RT-PCR and Sanger sequencing. Interestingly, novel_circ_003723, novel_circ_002714, novel_circ_002451, and novel_circ_001980 were highly expressed in both Ac1 and Ac2. In addition, the source genes of circRNAs were involved in 34 GO terms including organelle and cellular process and 141 pathways such as endocytosis and Wnt signaling pathway. Moreover, 55 DEcircRNAs including 34 upregulated and 21 downregulated circRNAs were identified in Ac2 compared with Ac1. circRNA-miRNA regulatory networks indicated that 1060 circRNAs can target 74 miRNAs; additionally, the DEcircRNA-miRNA-mRNA networks suggested that 13 downregulated circRNAs can bind to eight miRNAs and 29 miRNA-targeted mRNAs, while 16 upregulated circRNAs can link to 9 miRNAs and 29 miRNA-targeted mRNAs. These results indicated that DEcircRNAs as ceRNAs may play a comprehensive role in the growth, development, and metabolism of the worker's midgut via regulating source genes and interacting with miRNAs. Notably, eight DEcircRNAs targeting miR-6001-y were likely to be key participants in the midgut development. Our findings not only offer a valuable resource for further studies on A. cerana cerana circRNA and novel insights into understanding the molecular mechanisms underlying the midgut development of eastern honeybee but also provide putative circRNA candidates for functional research in the near future and novel biomarkers for identification of eastern honeybee species including A. cerana cerana and honeybee diseases such as chalkbrood and microsporidiosis.


Assuntos
Abelhas/genética , Trato Gastrointestinal/fisiologia , Redes Reguladoras de Genes , RNA Circular/genética , Animais , Biologia Computacional , Regulação para Baixo , MicroRNAs/genética , RNA Mensageiro/genética , Regulação para Cima
8.
Int J Mol Sci ; 21(5)2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32143291

RESUMO

Honey bee viruses are associated with honey bee colony decline. Israeli acute paralysis virus (IAPV) is considered to have a strong impact on honey bee survival. Phylogenetic analysis of the viral genomes from several regions of the world showed that various IAPV lineages had substantial differences in virulence. Chronic bee paralysis virus (CBPV), another important honey bee virus, can induce two significantly different symptoms. However, the infection characteristics and pathogenesis of IAPV and CBPV have not been completely elucidated. Here, we constructed infectious clones of IAPV and CBPV using a universal vector to provide a basis for studying their replication and pathogenesis. Infectious IAPV and CBPV were rescued from molecular clones of IAPV and CBPV genomes, respectively, that induced typical paralysis symptoms. The replication levels and expression proteins of IAPV and CBPV in progeny virus production were confirmed by qPCR and Western blot. Our results will allow further dissection of the role of each gene in the context of viral infection while helping to study viral pathogenesis and develop antiviral drugs using reverse genetics systems.


Assuntos
Abelhas/virologia , Dicistroviridae/genética , Vírus de Insetos/genética , Genética Reversa , Animais , Genoma Viral , Filogenia , Viroses/veterinária
9.
J Invertebr Pathol ; 166: 107210, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31211962

RESUMO

Chalkbrood is the most common fungal disease in honeybees. The objective of this study was to reveal immune responses in the Apis cerana cerana larval gut following Ascosphaera apis invasion. Combining a previously assembled transcriptome of A. c. cerana larval gut and the high-throughput sequencing data obtained in this study, 6152 differentially expressed genes (DEGs) were clustered into eight profiles. Trend analysis showed three significant up-regulated profiles (p ≤ 0.05) and three down-regulated profiles. Gene Ontology (GO) term analysis suggested that DEGs within significant up-regulated and down-regulated clusters were enriched in 46 and 38 functional groups, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis indicated a majority of DEGs were involved in ribosome structure or function, carbon metabolism, biosynthesis of amino acids, and oxidative phosphorylation. In addition, 142 and 14 DEGs were annotated in the cellular immune- and humoral immune-related pathways, respectively. Further investigation indicated that DEGs up-regulated in cellular immune and humoral immune pathways outnumbered those that were down-regulated. Moreover, immune responses of A. c. cerana and Apis mellifera ligustica larvae were compared and studied to decipher resistance of eastern honeybee larvae to A. apis. These results demonstrated that a large number of genes involved in immunity-related pathways were activated by A. apis. Our findings provided valuable information for elucidating the molecular mechanisms underlying immune responses of A. c. cerana larvae to A. apis infection and pathogen-host interactions during chalkbrood infection.


Assuntos
Abelhas/imunologia , Abelhas/microbiologia , Microbioma Gastrointestinal/imunologia , Proteínas de Insetos/imunologia , Micoses/veterinária , Animais , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno/imunologia , Onygenales/imunologia , Transcriptoma
10.
Curr Microbiol ; 75(12): 1655-1660, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30269253

RESUMO

Circular RNAs (circRNAs) are newly discovered endogenous non-coding RNAs (ncRNAs) that play key roles in microRNA function and transcriptional regulation. Though a large number of circRNAs had been identified in animals and plants, however, little is known regarding circRNAs in Nosema ceranae, a widespread fungal parasite of honeybee. In this study, using deep sequencing technology and bioinformatic analysis, we predicted 204 circRNAs from N. ceranae spore samples, including 174 exonic circRNAs and 30 intergenic circRNAs. In addition, the expression of seven N. ceranae circRNAs was confirmed by RT-PCR assay. Furthermore, regulation networks of circRNAs were constructed, and 15 circRNAs were found to act as sponges of the corresponding three miRNAs. GO categorization and pathway enrichment analysis suggested that the circRNAs are likely to play significant roles in N. ceranae spore. This is the first report of circRNAs generated by a microsporidia species. Our results provide novel insights into understanding the basic biology of N. ceranae.


Assuntos
DNA Fúngico/genética , Nosema/genética , Parasitos/genética , RNA/genética , Animais , Redes Reguladoras de Genes/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , MicroRNAs/genética , RNA Circular
11.
J Invertebr Pathol ; 156: 1-5, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29894727

RESUMO

Ascospheara apis is a widespread fungal pathogen that exclusively invades honeybee larvae. Thus far, non-coding RNA in A. apis has not yet been documented. In this study, we sequenced A. apis using strand specific cDNA library construction and Illumina RNA sequencing methods, and identified 379 lncRNAs, including antisense lncRNAs, lincRNAs, intronic lncRNAs and sense lncRNAs. Additionally, these lncRNAs were found to be shorter in length and have fewer exons and transcript isoforms than protein-coding genes, similar to those identified in mammals and plants. Furthermore, the existence of 15 predicted lncRNAs of A. apis was confirmed using RT-PCR and expression levels of 11 were lower than those of adjacent protein-coding genes. Our findings not only enlarge the lncRNA database for fungi, but also lay a foundation for further investigation of potential lncRNA-mediated regulation of genes in A. apis.


Assuntos
Fungos/genética , RNA Fúngico/genética , RNA Longo não Codificante/genética , Animais , Abelhas/parasitologia , RNA Fúngico/análise , RNA Longo não Codificante/análise
12.
Chin J Cancer Res ; 30(1): 84-92, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29545722

RESUMO

OBJECTIVE: Although laparoscopic treatment of gallbladder cancer (GBC) has been explored in the last decade, long-term results are still rare. This study evaluates long-term results of intended laparoscopic treatment for suspected GBC confined to the gallbladder wall, based on our experience over 10 years. METHODS: Between August 2006 and December 2015, 164 patients with suspected GBC confined to the wall were enrolled in the protocol for laparoscopic surgery. The process for GBC treatment was analyzed to evaluate the feasibility of computed tomography (CT) and/or magnetic resonance imaging (MRI) combined with frozen-section examination in identifying GBC confined to the wall. Of 159 patients who underwent the intended laparoscopic radical treatment, 47 with pathologically proven GBC were investigated to determine the safety and oncologic outcomes of a laparoscopic approach to GBC. RESULTS: Among the 164 patients, 5 patients avoided further radical surgery because of unresectable disease and 12 were converted to open surgery; in the remaining 147 patients, totally laparoscopic treatment was successfully accomplished. Extended cholecystectomy was performed in 37 patients and simple cholecystectomy in 10. The T stages based on final pathology were Tis (n=6), T1a (n=2), T1b (n=9), T2 (n=26), and T3 (n=4). Recurrence was detected in 11 patients over a median follow-up of 51 months. The disease-specific 5-year survival rate of these 47 patients was 68.8%, and rose to 85% for patients with a normal cancer antigen 19-9 (CA19-9) level. CONCLUSIONS: The favorable long-term outcomes demonstrate the feasibility of combined CT/MRI and frozen-section examination in the selection of patients with GBC confined to the gallbladder wall, confirm the oncologic safety of laparoscopic treatment in selected GBC patients, and favor measurement of preoperative CA19-9 in the selection of GBCs suitable for laparoscopic treatment.

13.
J Gen Virol ; 98(11): 2864-2875, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29058655

RESUMO

By providing pollination services, bees are among the most important insects, both in ecological and economical terms. Combined next-generation and classical sequencing approaches were applied to discover and study new insect viruses potentially harmful to bees. A bioinformatics virus discovery pipeline was used on individual Illumina transcriptomes of 13 wild bees from three species from the genus Halictus and 30 ants from six species of the genera Messor and Aphaenogaster. This allowed the discovery and description of three sequences of a new virus termed Halictus scabiosae Adlikon virus (HsAV). Phylogenetic analyses of ORF1, RNA-dependent RNA-polymerase (RdRp) and capsid genes showed that HsAV is closely related to (+)ssRNA viruses of the unassigned Sinaivirus genus but distant enough to belong to a different new genus we called Halictivirus. In addition, our study of ant transcriptomes revealed the first four sinaivirus sequences from ants (Messor barbarus, M. capitatus and M. concolor). Maximum likelihood phylogenetic analyses were performed on a 594 nt fragment of the ORF1/RdRp region from 84 sinaivirus sequences, including 31 new Lake Sinai viruses (LSVs) from honey bees collected in five countries across the globe and the four ant viral sequences. The phylogeny revealed four main clades potentially representing different viral species infecting honey bees. Moreover, the ant viruses belonged to the LSV4 clade, suggesting a possible cross-species transmission between bees and ants. Lastly, wide honey bee screening showed that all four LSV clades have worldwide distributions with no obvious geographical segregation.


Assuntos
Formigas/virologia , Abelhas/virologia , Vírus de Insetos/classificação , Vírus de Insetos/isolamento & purificação , Filogenia , Animais , Variação Genética , Vírus de Insetos/genética , Análise de Sequência de DNA , Proteínas Virais/genética
14.
Arch Virol ; 162(8): 2451-2456, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28432563

RESUMO

Chronic bee paralysis virus (CBPV) is an important viral pathogen that affects adult bees. Although several CBPV strains have been reported, little information has been obtained from China. In this study, two major segments of the CBPV Chinese isolate CBPV-BJ, RNA 1 and RNA 2, were determined to be 3657 and 2267 nucleotides (nt) in length, respectively. RNA 1 and RNA 2 contained three and four open reading frames (ORFs), respectively, which agreed with known reference strains (EU122229 and EU122230). The RNA 1 had 98% nucleotide sequence identity to a known Chinese strain (KU950353), and RNA 2 had 97% nucleotide sequence identity to another Chinese strain (KU950354). Although the lengths of the RNA 1 and RNA 2 sequences were 17 nt and 38 nt shorter than those of the CBPV reference strains EU122229 and EU122230, respectively, the complete CBPV-BJ RNA 1 and RNA 2 sequences shared 91% and 92% identity with them. Phylogenetic analysis based on the sequences of the RNA-dependent RNA polymerase (RdRp) and putative structural proteins (pSPs) showed that CBPV-BJ was most closely related to the other two Chinese isolate (KU950353 and KU950354) and clustered with most Asian strains. These data provide new information that will lead to a better understanding of the diversity of the CBPV genome.


Assuntos
Abelhas/virologia , Vírus de Insetos/genética , Vírus de RNA/genética , RNA Viral/genética , Animais , China , Variação Genética , Genoma Viral , Vírus de Insetos/isolamento & purificação , Vírus de Insetos/patogenicidade , Fases de Leitura Aberta , Vírus de RNA/isolamento & purificação , Análise de Sequência de RNA
15.
J Invertebr Pathol ; 138: 112-5, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27369386

RESUMO

Although many honey bee RNA viruses have been correlated with colony declines, little is known regarding the potential role of DNA viruses. Here, we examined seemingly healthy and crawling bee samples from China using PCR to identify whether Apis mellifera filamentous virus (AmFV) was present in A. cerana cerana. The highest AmFV infection percentage among Chinese provinces occurred in crawling bees from Gansu province (85.48%), and the lowest was in bees from Beijing (31.58%). A phylogenetic analysis showed that the Chinese isolate of AmFV exhibited a high genetic similarity with isolates from Belgium, Switzerland and USA. This is the first report of AmFV infections in Chinese A. cerana cerana populations.


Assuntos
Abelhas/virologia , Viroses/veterinária , Animais , China , Vírus de DNA , Genes Virais/genética , Filogenia , Reação em Cadeia da Polimerase
17.
Mol Biol Rep ; 41(12): 8081-91, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25189650

RESUMO

Honey bee is important economic insect that not only pollinates fruits and crops but also provides products with various physiological activities. Bee venom is a functional agent that is widely applied in clinical treatment and pharmacy. Secapin is one of these agents that have a significant role in therapy. The functions of secapin from the bee venom have been documented, but little information is known about its heterologous expression under natural condition. Moreover, few scholars verified experimentally the functions of secapin from bee venom in vitro. In this study, we successfully constructed a heterologous expression vector, which is different from conventional expression system. A transgenic approach was established for transformation of secapin gene from the venom of Apis mellifera carnica (Ac-sec) into the edible fungi, Coprinus cinereus. Ac-sec was encoded by a 234 bp nucleotide that contained a signal peptide domain and two potential phosphorylation sites. The sequence exhibited highly homology with various secapins characterized from honey bee and related species. Southern blot data indicated that Ac-sec was present as single or multiple copy loci in the C. cinereus genome. By co-transformation and double-layer active assay, Ac-sec was expressed successfully in C. cinereus and the antibacterial activity of the recombinants was identified, showing notable antibacterial activities on different bacteria. Although Ac-sec is from the venom of Apidae, phylogenetic analysis demonstrated that Ac-sec was more closely related to that of Vespid than to bee species from Apidae. The molecular characteristics of Ac-sec and the potential roles of small peptides in biology were discussed.


Assuntos
Antibacterianos , Venenos de Abelha/genética , Coprinus/genética , Sequência de Aminoácidos , Animais , Abelhas , Clonagem Molecular , Dados de Sequência Molecular , Organismos Geneticamente Modificados , Filogenia , Proteínas Recombinantes/biossíntese , Alinhamento de Sequência , Transgenes
18.
ISME J ; 18(1)2024 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-38519112

RESUMO

The significance of gut microbiota in regulating animal immune response to viral infection is increasingly recognized. However, how chronic bee paralysis virus (CBPV) exploits host immune to disturb microbiota for its proliferation remains elusive. Through histopathological examination, we discovered that the hindgut harbored the highest level of CBPV, and displayed visible signs of damages. The metagenomic analysis showed that a notable reduction in the levels of Snodgrassella alvi and Lactobacillus apis, and a significant increase in the abundance of the opportunistic pathogens such as Enterobacter hormaechei and Enterobacter cloacae following CBPV infection. Subsequent co-inoculation experiments showed that these opportunistic pathogens facilitated the CBPV proliferation, leading to accelerated mortality in bees and exacerbation of bloated abdomen symptoms after CBPV infection. The expression level of antimicrobial peptide (AMP) was found to be significantly up-regulated by over 1000 times in response to CBPV infection, as demonstrated by subsequent transcriptome and quantitative real-time PCR investigations. In particular, through correlation analysis and a bacteriostatic test revealed that the AMPs did not exhibit any inhibitory effect against the two opportunistic pathogens. However, they did demonstrate inhibitory activity against S. alvi and L. apis. Our findings provide different evidence that the virus infection may stimulate and utilize the host's AMPs to eradicate probiotic species and facilitate the proliferation of opportunistic bacteria. This process weakens the intestinal barrier and ultimately resulting in the typical bloated abdomen.


Assuntos
Microbioma Gastrointestinal , Vírus de Insetos , Vírus de RNA , Viroses , Vírus , Abelhas , Animais , Vírus de RNA/fisiologia , Peptídeos Antimicrobianos , Vírus de Insetos/fisiologia , Paralisia
19.
Int J Biol Macromol ; 269(Pt 2): 132103, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38719011

RESUMO

Rhodotorula spp. has been studied as one powerful source for a novel cell factory with fast growth and its high added-value biomolecules. However, its inadequate genome and genomic annotation have hindered its widespread use in cosmetics and food industries. Rhodotorula glutinis QYH-2023, was isolated from rice rhizosphere soil, and the highest quality of the genome of the strain was obtained at chromosome level (18 chromosomes) than ever before in red yeast in this study. Comparative genomics analysis revealed that there are more key gene copies of carotenoids biosynthesis in R. glutinis QYH-2023 than other species of Rhodotorula spp. Integrated transcriptome and metabolome analysis revealed that lipids and carotenoids biosynthesis was significantly enriched during fermentation. Subsequent investigation revealed that the over-expression of the strain three genes related to carotenoids biosynthesis in Komagataella phaffii significantly promoted the carotenoid production. Furthermore, in vitro tests initially confirmed that the longer the fermentation period, the synthesized metabolites controlled by R. glutinis QYH-2023 genome had the stronger anti-inflammatory properties. All of the findings revealed a high-quality reference genome which highlight the potential of R. glutinis strains to be employed as chassis cells for biosynthesizing carotenoids and other active chemicals.


Assuntos
Carotenoides , Genoma Fúngico , Rhodotorula , Carotenoides/metabolismo , Rhodotorula/genética , Rhodotorula/metabolismo , Anti-Inflamatórios/farmacologia , Fermentação , Cromossomos Fúngicos/genética , Genômica/métodos , Transcriptoma
20.
Clin Cosmet Investig Dermatol ; 16: 3023-3034, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37901151

RESUMO

Objective: High levels of VEGF and excessive angiogenesis contribute significantly to hypertrophic scar (HS) formation. Our study aimed to preliminarily investigate the effect of axitinib, a selective VEGF receptor tyrosine kinase inhibitor, on angiogenesis of HS and to explore its possible mechanism in a rabbit ear model. Methods: Ten male New Zealand white rabbits were used to establish HS models and then randomised to the control and axitinib groups. The scar tissues in the two groups were injected with axitinib or normal saline, and they were evaluated after one month of treatment. Macroscopic scar thickness, vascularity and pliability, as well as histopathological analysis including HE staining and Masson staining and scar elevation index (SEI) between two groups were compared. Immunohistochemical staining of CD31 in two groups was conducted to assess the degree of angiogenesis in HS tissue. The protein expression of protein kinase B (AKT) and ribosomal protein S6 kinase (p70S6K) and their phosphorylation levels in both groups were examined by Western blot analysis. Results: The macroscopic and histological observation showed intralesional axitinib injection significantly reduced scar thickness, vascularity and pliability of HS in the rabbit ear model. The value of SEI in HE assessment was also significantly declined in the axitinib group. Furthermore, immunohistochemical analysis revealed that axitinib suppressed the expression of CD31 in HS tissue, and the mean IOD for blood vessels was significantly lower in the axitinib-treated group. Additionally, axitinib effectively attenuated the protein expression of p70S6K, p-AKT and p-p70S6K by Western blot analysis. Conclusion: Our study suggests that intralesional injection of axitinib can effectively attenuate HS by reducing angiogenesis in the rabbit ear model, and this inhibitory effect may be mediated by suppression of AKT/p70S6K signaling pathway. It indicates that axitinib may be a promising option for the treatment of HS in the future.

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