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1.
Ann Plast Surg ; 90(5S Suppl 2): S183-S186, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-36752413

RESUMO

OBJECTIVE: To investigate the application of dual "kite" myomucosal flaps (subcutaneous pedicle advancement flap) for the repair of medium lip defects (one-third to one-half lip width). METHODS: Dual kite myomucosal flaps were designed in the adjacent area of the defect in 17 patients with medium lip defect with the principle of using homogenous tissue as far as possible without affecting local anatomical units. RESULTS: The follow-up time was 3 to 24 months; 16 patients showed primary wound healing, and 1 patient showed prolonged healing. The blood supply of the myomucosal flaps were reliable. The myomucosal flaps were smooth, with no proliferation of scars, and the local appearance was good. CONCLUSION: The dual kite myomucosal flaps provide a reliable method for repairing medium lip defects, decreasing the need for additional excision of normal skin tissue, and reducing skin scar.


Assuntos
Neoplasias Labiais , Procedimentos de Cirurgia Plástica , Lesões dos Tecidos Moles , Humanos , Lábio/cirurgia , Retalhos Cirúrgicos/cirurgia , Neoplasias Labiais/cirurgia , Cicatriz/prevenção & controle , Cicatriz/cirurgia , Transplante de Pele , Resultado do Tratamento , Lesões dos Tecidos Moles/cirurgia
2.
Ann Plast Surg ; 90(5S Suppl 2): S230-S233, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-36752400

RESUMO

OBJECTIVE: The aim of this study was to analyze the epidemiological characteristics of pediatric facial soft tissue injuries of Chinese preschool-aged children in Hangzhou Plastic Surgery Hospital. METHODS: Medical records of preschool-aged children's facial injuries, 6 years and younger, from January 2017 to December 2019 were collected. Sex; age; time of injury; length of stay; causes of injury; location, type, length, and depth of wound; anesthesia methods; and treatment and evaluation of postoperative scars were analyzed. RESULTS: There were 10,862 cases (male, 6780 cases; female, 4082 cases) in the group. The ratio of male to female was 1.66:1. Mean age was 3.4 (±1.6) years; the youngest was 1 month old. The time of injury occurred frequently between 9:00 and 13:00 and 16:00 to 21:00, with the most common incident time being between 19:00 and 20:00. Collision injury was the main cause of injury (9822 [90.43%]). The most frequently injured area was the forehead (4874 [44.87%]). The main form of injury was laceration wound (9721 [89.45%]). The depth of injuries was mainly middle layer (adipose or muscular layer) (6299 [57.99%]). The length of injuries was 1.7 (±0.9) cm, ranging from 0.2 to 10.5 cm. Furthermore, 9110 cases were repaired by plastic surgeries and 1 or more antiscar measures. After 6-month to 2-year follow-up, 9 cases of animal scratch or bite, lip penetrating wound, or bumping teeth were infected and 26 cases had scar hyperplasia. The others achieved satisfactory results, and the scars were not obvious. CONCLUSION: Preschool-aged children's facial injuries have predictable patterns of occurrence, and targeted preventive measures can reduce the incidence rates. After facial injury, children should present for timely plastic surgery treatment and accept combined antiscarring measures to minimize postoperative scarring.


Assuntos
Traumatismos Faciais , Procedimentos de Cirurgia Plástica , Lesões dos Tecidos Moles , Feminino , Humanos , Masculino , Cicatriz/etiologia , Cicatriz/complicações , População do Leste Asiático , Traumatismos Faciais/epidemiologia , Traumatismos Faciais/etiologia , Traumatismos Faciais/cirurgia , Lesões dos Tecidos Moles/epidemiologia , Lesões dos Tecidos Moles/etiologia , Lesões dos Tecidos Moles/cirurgia , Lactente , Pré-Escolar
3.
Ann Plast Surg ; 90(5S Suppl 2): S139-S142, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-36752499

RESUMO

ABSTRACT: Asian nasal characteristics include a low dorsum, short nose, and thick skin envelope, usually requiring lengthening and elevating the nose during rhinoplasty ( Facial Plast Surg Clin North Am 2007;15(3):293-307, v). The increase in rhinoplasty popularity has resulted in a greater prevalence of complications. In a severely short and contracted nose, an extensively scarred or contracted soft tissue envelope results in weak laxity and extensibility of the nasal skin. For these patients, the essential component of rhinoplasty is to improve skin texture and obtain a sufficient nasal skin soft tissue envelope. Tissue expanders have previously been utilized to expand nasal skin and soft tissue ( Plast Reconstr Surg 2006;118(6):1447-1452; Facial Plast Surg 2019;35(1):68-72). However, nasal anatomical characteristics have limited the clinical application of tissue expanders. This article introduces a simple, noninvasive, and easily adopted method of external nasal skin stretching, which was first proposed by the senior author. This approach has been accepted by rhinoplasty surgeons in China and widely used in clinic. The approach can improve skin laxity, yield extra nasal soft tissue, and create adequate soft tissue coverage of the reconstructed nasal framework to reduce the difficulty of surgery with a reliable clinical effect.


Assuntos
Doenças Nasais , Rinoplastia , Humanos , Rinoplastia/métodos , Nariz/cirurgia , Pele , Doenças Nasais/cirurgia , Cicatriz/cirurgia
4.
J Craniofac Surg ; 34(6): 1661-1665, 2023 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-37220717

RESUMO

Rhinoplasty is a common surgical procedure in medical cosmetology. From patients with saddle nose deformity to beauty seekers with low and short noses, this surgery is mainly sought to improve the nose's appearance. To investigate the effect of modified auricular cartilage scaffold combined with L-shaped prosthesis in rhinoplasty. This retrospective study included 54 patients who underwent auricular cartilage augmentation rhinoplasty with L-shaped implants in our hospital from July 2018 to July 2021. The function of nasal ventilation and olfaction was inspected. As a result, the degree of nasal tip protrusion and the changes in the superior lip angle of columella were improved. The patients' satisfaction was measured a year after the surgery. Patients who underwent auricular cartilage augmentation rhinoplasty with L-shaped prosthesis were satisfied with the surgery outcomes. Using a protective auricular cartilage scaffold combined with an L-shaped implant for augmentation rhinoplasty reduced the shortage of the application and reinforced the stability of the auricular cartilage augmentation rhinoplasty. At >12 months follow-up, there were no serious adverse effects on nasal ventilation and olfactory function in any of the patients. The presented method made full use of auricular cartilage so that it reduced the harvest of the cartilage. Besides, it achieved the remarkable lift of the nose tip, thus simulating the appearance of costal cartilage rhinoplasty. Furthermore, the risk of implant exposure was efficiently reduced, making it worthy of clinical application.


Assuntos
Cartilagem Costal , Implantes Dentários , Doenças Nasais , Rinoplastia , Humanos , Rinoplastia/métodos , Cartilagem da Orelha/cirurgia , Estudos Retrospectivos , Nariz/cirurgia , Septo Nasal/cirurgia , Cartilagem Costal/cirurgia , Doenças Nasais/cirurgia
5.
Angew Chem Int Ed Engl ; 62(22): e202301074, 2023 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-36961095

RESUMO

The development of PtIV prodrugs that are reduced into the therapeutically active PtII species within the tumor microenvironment has received much research interest. In order to provide spatial and temporal control over the treatment, there is a high demand for the development of compounds that could be selectively activated upon irradiation. Despite recent progress, the majority of PtIV complexes are excited with ultraviolet or blue light, limiting the use of such compounds to superficial application. To overcome this limitation, herein, the first example of PtIV prodrug nanoparticles that could be reduced with deeply penetrating ultrasound radiation is reported, enabling the treatment of deep-seated or large tumors. The nanoparticles were found to selectively accumulate inside a mouse colon carcinoma tumor upon intravenous injection and were able to eradicate the tumor upon exposure to ultrasound radiation.


Assuntos
Antineoplásicos , Nanopartículas , Neoplasias , Pró-Fármacos , Animais , Camundongos , Pró-Fármacos/farmacologia , Pró-Fármacos/uso terapêutico , Platina/uso terapêutico , Neoplasias/tratamento farmacológico , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Microambiente Tumoral
6.
Small ; 18(1): e2104624, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34761500

RESUMO

Alkaline water electrolysis is an advanced technology for scalable H2 production using surplus electricity from intermittent energy sources, but it remains challenging for non-noble electrocatalysts to split water into hydrogen and oxygen efficiently, especially for tungsten disulfide (WS2 )-based catalysts. Density functional theory calculations in combination with experimental study are used to establish a multi-site engineering strategy for developing robust WS2 -based hybrid electrocatalyst on mesoporous bimetallic nitride (Ni3 FeN) nanoarrays for bifunctional water splitting. This ingenious design endows the catalyst with numerous edge sites chemically bonded with the conductive scaffold, which are favorable for water dissociation and hydrogen adsorption. Benefiting from the synergistic advantages, the N-WS2 /Ni3 FeN hybrid exhibits exceptional bifunctional properties for hydrogen and oxygen evolution reactions (HER and OER) in base with excellent large-current durability, requiring 84 mV to afford 10 mA cm-2 for HER, and 240 mV at 100 mA cm-2 for OER, respectively. Assembling the catalytic materials as both the anode and cathode to construct an electrolyzer, it is actualized very good activities for overall water splitting with only 1.5 V to deliver 10 mA cm-2 , outperforming the IrO2 (+) //Pt(-) coupled electrodes and many non-noble bifunctional electrocatalysts thus far. This work provides a promising avenue for designing WS2 -based heterogeneous electrocatalysts for water electrolysis.

7.
Small ; 18(4): e2105642, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34825490

RESUMO

The catalytic hydrogen-evolving activities of transition-metal phosphides are greatly related to the phosphorus content, but the physical origin of performance enhancement remains ambiguous, and tuning the catalytic activity of nickel phosphides (NiP2 /Ni5 P4 ) remains challenging due to unfavorable H* adsorption. Here, a strategy is introduced to integrate P-rich NiP2 and P-poor Ni5 P4 into in-plane heterostructures by anion substitution, in which P atoms at the in-plane interfaces perform as active sites to adsorb H* and thus facilitate the hydrogen evolution reaction (HER) process via modulating the electronic structure between NiP2 and Ni5 P4 . Consequently, the NiP2 /Ni5 P4 hybrid exhibits an outstanding hydrogen-evolving activity, requiring only 30 and 76 mV to afford 10 and 100 mA cm-2 in acid, respectively. It surpasses most of the earth-abundant electrocatalysts thus far, and is comparable to Pt catalysts (30/72 mV at 10/100 mA cm-2 ). Particularly, it can run smoothly at large current density and only requires 247 mV to reach 2000 mA cm-2 . Detailed theoretical calculations reveal that its exceptional activity stems from the moderate overlap of density states between P 2p and H 1s orbitals, thus optimizing the H*-adsorption strength. This work highlights a new avenue toward the fabrication of robust non-noble electrocatalysts by constructing in-plane heterojunctions.

8.
Biotechnol Lett ; 44(1): 59-76, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34997407

RESUMO

Gene-knockout pigs have important applications in agriculture and medicine. Compared with CRISPR/Cas9, Adenine base editor (ABE) convert single A·T pairs to G·C pairs in the genome without generating DNA double-strand breaks, and this method has higher accuracy and biosafety in pig genetic modification. However, the application of ABE in pig gene knockout is limited by protospacer-adjacent motif sequences and the base-editing window. Alternative mRNA splicing is an important mechanism underlying the formation of proteins with diverse functions in eukaryotes. Spliceosome recognizes the conservative sequences of splice donors and acceptors in a precursor mRNA. Mutations in these conservative sequences induce exon skipping, leading to proteins with novel functions or to gene inactivation due to frameshift mutations. In this study, adenine base-editing-mediated exon skipping was used to expand the application of ABE in the generation of gene knockout pigs. We first constructed a modified "all-in-one" ABE vector suitable for porcine somatic cell transfection that contained an ABE for single-base editing and an sgRNA expression cassette. The "all-in-one" ABE vector induced efficient sgRNA-dependent A-to-G conversions in porcine cells during single base-editing of multiple endogenous gene loci. Subsequently, an ABE system was designed for single adenine editing of the conservative splice acceptor site (AG sequence at the 3' end of the intron 5) and splice donor site (GT sequence at the 5' end of the intron 6) in the porcine gene GHR; this method achieved highly efficient A-to-G conversion at the cellular level. Then, porcine single-cell colonies carrying a biallelic A-to-G conversion in the splice acceptor site in the intron 5 of GHR were generated. RT-PCR indicated exon 6 skipped at the mRNA level. Western blotting revealed GHR protein loss, and gene sequencing showed no sgRNA-dependent off-target effects. These results demonstrate accurate adenine base-editing-mediated exon skipping and gene knockout in porcine cells. This is the first proof-of-concept study of adenine base-editing-mediated exon skipping for gene regulation in pigs, and this work provides a new strategy for accurate and safe genetic modification of pigs for agricultural and medical applications.


Assuntos
Adenina , Edição de Genes , Adenina/metabolismo , Animais , Sistemas CRISPR-Cas/genética , Linhagem Celular , Éxons/genética , Edição de Genes/métodos , Técnicas de Inativação de Genes , Suínos
9.
Angew Chem Int Ed Engl ; 61(31): e202203546, 2022 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-35642869

RESUMO

Recent progress in studying copper-dependent targets and pathways in the context of tumor treatment has provided new insights into therapeutic strategies of leveraging copper-dependent disease vulnerabilities and pharmacological manipulation of intratumor copper transportation to improve chemotherapy. Here, we developed reactive oxygen species (ROS)-sensitive nanoparticles loaded with copper chaperone inhibitor DC_AC50 and cisplatin(IV) prodrug. The released DC_AC50 can promote a remarkable accumulation of intracellular cisplatin and copper through inhibition of the Atox1-ATPase pathways, thereby enhancing the chemotherapeutic effect of cisplatin and inducing significant ROS generation. Excessive ROS then elicits intense endoplasmic reticulin (ER) stress which facilitates the immunogenic cell death (ICD) spurring a sustained immune response. Our study suggests that nanoparticle-mediated copper chaperone inhibition via DC_AC50 can restore the immunogenicity of tumor cells for enhanced chemotherapy and cancer immunotherapy.


Assuntos
Nanopartículas , Neoplasias , Linhagem Celular Tumoral , Cisplatino/metabolismo , Cisplatino/farmacologia , Cobre/metabolismo , Chaperonas Moleculares , Neoplasias/tratamento farmacológico , Espécies Reativas de Oxigênio/metabolismo
10.
BMC Genomics ; 22(1): 199, 2021 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-33745451

RESUMO

BACKGROUND: The red swamp crayfish Procambarus clarkii is a freshwater species that possesses high adaptability, environmental tolerance, and fecundity. P. clarkii is artificially farmed on a large scale in China. However, the molecular mechanisms of ovarian development in P. clarkii remain largely unknown. In this study, we identified four stages of P. clarkii ovary development, the previtellogenic stage (stage I), early vitellogenic stage (stage II), middle vitellogenic stage (stage III), and mature stage (stage IV) and compared the transcriptomics among these four stages through next-generation sequencing (NGS). RESULTS: The total numbers of clean reads of the four stages ranged from 42,013,648 to 62,220,956. A total of 216,444 unigenes were obtained, and the GC content of most unigenes was slightly less than the AT content. Principal Component Analysis (PCA) and Anosim analysis demonstrated that the grouping of these four stages was feasible, and each stage could be distinguished from the others. In the expression pattern analysis, 2301 genes were continuously increase from stage I to stage IV, and 2660 genes were sharply decrease at stage IV compared to stages I-III. By comparing each of the stages at the same time, four clusters of differentially expressed genes (DEGs) were found to be uniquely highly expressed in stage I (136 genes), stage II (43 genes), stage III-IV (49 genes), and stage IV (22 genes), thus exhibiting developmental stage specificity. Moreover, in comparisons between adjacent stages, the number of DEGs between stage III and IV was the highest. GO enrichment analysis demonstrated that nutrient reservoir activity was highest at stage II and that this played a foreshadowing role in ovarian development, and the GO terms of cell, intracellular and organelle participated in the ovary maturation during later stages. In addition, KEGG pathway analysis revealed that the early development of the ovary was mainly associated with the PI3K-Akt signaling pathway and focal adhesion; the middle developmental period was related to apoptosis, lysine biosynthesis, and the NF-kappa B signaling pathway; the late developmental period was involved with the cell cycle and the p53 signaling pathway. CONCLUSION: These transcriptomic data provide insights into the molecular mechanisms of ovarian development in P. clarkii. The results will be helpful for improving the reproduction and development of this aquatic species.


Assuntos
Astacoidea , Transcriptoma , Animais , Astacoidea/genética , China , Feminino , Perfilação da Expressão Gênica , Ovário , Fosfatidilinositol 3-Quinases
11.
Nanotechnology ; 33(10)2021 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-34847544

RESUMO

The transport and storage of ions within solid state structures is a fundamental limitation for fabricate more advanced electrochemical energy storage, memristor, and electrochromic devices. Crystallographic shear structure can be induced in the tungsten bronze structures composed of corner-sharing WO6octahedra by the addition of edge-sharing NbO6octahedra, which might provide more storage sites and more convenient transport channels for external ions such as hydrogen ions and alkali metal ions. Here, we show that Nb2O5·15WO3nanowires (Nb/W = 0.008) with long length-diameter ratio, smooth surface, and uniform diameter have been successfully synthesized by a simple hydrothermal method. The Nb2O5·15WO3nanowires do exhibit more advantages over h-WO3nanowires in electrochemical hydrogen ion storage such as smaller polarization, larger capacity (71 mAh g-1, at 10C, 1C = 100 mA g-1), better cycle performance (remain at 99% of the initial capacity after 200 cycles at 100C) and faster H+ions diffusion kinetics. It might be the crystallographic shear structure induced by Nb doping that does result in the marked improvement in the hydrogen-ion storage performance of WO3. Therefore, complex niobium tungsten oxide nanowires might offer great promise for the next generation of electrochemical energy and information storage devices.

12.
Biotechnol Lett ; 43(9): 1699-1714, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34189671

RESUMO

Bama minipig is a unique miniature swine bred from China. Their favorable characteristics include delicious meat, strong adaptability, tolerance to rough feed, and high levels of stress tolerance. Unfavorable characteristics are their low lean meat percentage, high fat content, slow growth rate, and low feed conversion ratio. Genome-editing technology using CRISPR/Cas9 efficiently knocked out the myostatin gene (MSTN) that has a negative regulatory effect on muscle production, effectively promoting pig muscle growth and increasing lean meat percentage of the pigs. However, CRISPR/Cas9 genome editing technology is based on random mutations implemented by DNA double-strand breaks, which may trigger genomic off-target effects and chromosomal rearrangements. The application of CRISPR/Cas9 to improve economic traits in pigs has raised biosafety concerns. Base editor (BE) developed based on CRISPR/Cas9 such as cytosine base editor (CBE) effectively achieve targeted modification of a single base without relying on DNA double-strand breaks. Hence, the method has greater safety in the genetic improvement of pigs. The aim of the present study is to utilize a modified CBE to generate MSTN-knockout cells of Bama minipigs. Our results showed that the constructed "all-in-one"-modified CBE plasmid achieved directional conversion of a single C·G base pair to a T·A base pair of the MSTN target in Bama miniature pig fibroblast cells. We successfully constructed multiple single-cell colonies of Bama minipigs fibroblast cells carrying the MSTN premature termination and verified that there were no genomic off-target effects detected. This study provides a foundation for further application of somatic cell cloning to construct MSTN-edited Bama minipigs that carry only a single-base mutation and avoids biosafety risks to a large extent, thereby providing experience and a reference for the base editing of other genetic loci in Bama minipigs.


Assuntos
Citosina/metabolismo , Fibroblastos/citologia , Edição de Genes/métodos , Miostatina/genética , Animais , Proteína 9 Associada à CRISPR/metabolismo , Sistemas CRISPR-Cas , Células Cultivadas , Códon de Terminação , Fibroblastos/metabolismo , Plasmídeos/genética , Suínos , Porco Miniatura , Transfecção
13.
Small ; 16(13): e1906629, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32133786

RESUMO

Developing efficient non-noble and earth-abundant hydrogen-evolving electrocatalysts is highly desirable for improving the energy efficiency of water splitting in base. Molybdenum disulfide (MoS2 ) is a promising candidate, but its catalytic activity is kinetically retarded in alkaline media due to the unfavorable water adsorption and dissociation feature. A heterogeneous electrocatalyst is reported that is constructed by selenium-doped MoS2 (Se-MoS2 ) particles on 3D interwoven cobalt diselenide (CoSe2 ) nanowire arrays that drives the hydrogen evolution reaction (HER) with fast reaction kinetics in base. The resultant Se-MoS2 /CoSe2 hybrid exhibits an outstanding catalytic HER performance with extremely low overpotentials of 30 and 93 mV at 10 and 100 mA cm-2 in base, respectively, which outperforms most of the inexpensive alkaline HER catalysts, and is among the best alkaline catalytic activity reported so far. Moreover, this hybrid catalyst shows exceptional catalytic performance with very low overpotentials of 84 and 95 mV at 10 mA cm-2 in acidic and neutral electrolytes, respectively, implying robust pH universality of this hybrid catalyst. This work may provide new inspirations for the development of high-performance MoS2 -based HER electrocatalysts in unfavorable basic media for promising catalytic applications.

14.
Biotechnol Lett ; 42(11): 2091-2109, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32494996

RESUMO

OBJECTIVES: Guangdong Small-ear Spotted (GDSS) pigs are a pig breed native to China that possesses unfortunate disadvantages, such as slow growth rate, low lean-meat percentage, and reduced feed utilization. In contrast to traditional genetic breeding methods with long cycle time and high cost, CRISPR/Cas9-mediated gene editing for the modification of the pig genome can quickly improve production traits, and therefore this technique exhibits important potential in the genetic improvement and resource development of GDSS pigs. In the present study, we aimed to establish an efficient CRISPR/Cas9-mediated gene-editing system for GDSS pig cells by optimizing the electrotransfection parameters, and to realize efficient CRISPR/Cas9-mediated gene editing of GDSS pig cells. RESULTS: After optimization of electrotransfection parameters for the transfection of GDSS pig cells, we demonstrated that a voltage of 150 V and a single pulse with a pulse duration of 20 ms were the optimal electrotransfection parameters for gene editing in these cells. In addition, our study generated GDSS pig single-cell colonies with biallelic mutations in the myostatin (MSTN) gene and insulin-like growth factor 2 (IGF2) intron-3 locus, which play an important role in pig muscle growth and muscle development. The single-cell colonies showed no foreign gene integration or off-target effects, and maintained normal cell morphology and viability. These gene-edited, single-cell colonies can in the future be used as donor cells to generate MSTN- and IGF2-edited GDSS pigs using somatic cell nuclear transfer (SCNT). CONCLUSIONS: This study establishes the foundation for genetic improvement and resource development of GDSS pigs using CRISPR/Cas9-mediated gene editing combined with SCNT.


Assuntos
Edição de Genes/métodos , Fator de Crescimento Insulin-Like II/genética , Miostatina/genética , Transfecção/métodos , Animais , Sistemas CRISPR-Cas , Linhagem Celular , Fenômenos Eletromagnéticos , Mutação , Seleção Artificial , Análise de Célula Única , Suínos
15.
Reprod Domest Anim ; 55(10): 1314-1327, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32679613

RESUMO

CRISPR/Cas9-mediated genome editing technology is a simple and highly efficient and specific genome modification approach with wide applications in the animal industry. CRISPR/Cas9-mediated genome editing combined with somatic cell nuclear transfer rapidly constructs gene-edited somatic cell-cloned pigs for the genetic improvement of traits or simulation of human diseases. Chinese Bama pigs are an excellent indigenous minipig breed from Bama County of China. Research on genome editing of Chinese Bama pigs is of great significance in protecting its genetic resource, improving genetic traits and in creating disease models. This study aimed to address the disadvantages of slow growth and low percentage of lean meat in Chinese Bama pigs and to knock out the myostatin gene (MSTN) by genome editing to promote growth and increase lean meat production. We first used CRISPR/Cas9-mediated genome editing to conduct biallelic knockout of the MSTN, followed by somatic cell nuclear transfer to successfully generate MSTN biallelic knockout Chinese Bama pigs, which was confirmed to have significantly faster growth rate and showed myofibre hyperplasia when they reached sexual maturity. This study lays the foundation for the rapid improvement of production traits of Chinese Bama pigs and the generation of gene-edited disease models in this breed.


Assuntos
Sistemas CRISPR-Cas , Miostatina/genética , Porco Miniatura/genética , Animais , Feminino , Técnicas de Inativação de Genes/veterinária , Masculino , Fibras Musculares Esqueléticas/fisiologia , Técnicas de Transferência Nuclear/veterinária , Carne de Porco , Suínos , Porco Miniatura/crescimento & desenvolvimento
16.
Int J Mol Sci ; 21(2)2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31941098

RESUMO

Lysozymes are key antimicrobial peptides in the host innate immune system that protect against pathogen infection. In this study, the full-length cDNAs of two c-type lysozymes (gfLyz-C1 and gfLyz-C2) were cloned from goldfish (Carassius auratus). The structural domains, three-dimensional structures, and amino acid sequences of gfLyz-C1 and gfLyz-C2 were highly comparable, as the two proteins shared 89.7% sequence identity. The gfLyz-C1 and gfLyz-C2 recombinant proteins were generated in the insoluble fractions of an Escherichia coli system. Based on the results of lysoplate and turbidimetric assays, gfLyz-C1 and gfLyz-C2 showed broad-spectrum antimicrobial properties with high levels of activity against Micrococcus lysodeikticus, Vibrio parahemolyticus, and Edwardsiella tarda, and relatively low activity against E. coli. Both gfLyz-C1 and gfLyz-C2 mRNAs were mainly expressed in the trunk kidney and head kidney, and gfLyz-C1 was expressed at much higher levels than gfLyz-C2 in the corresponding tissues. The expression of the gfLyz-C1 and gfLyz-C2 transcripts in the trunk kidney and head kidney was induced in these tissues by challenge with heat-inactivated E. coli and lipopolysaccharides (LPS), and the transcriptional responses of gfLyz-C1 were more intense. In goldfish primary trunk kidney cells, the levels of the gfLyz-C1 and gfLyz-C2 transcripts were upregulated by heat-inactivated E. coli, V. parahemolyticus, and E. tarda, as well as LPS, and downregulated by treatment with dexamethasone and leptins. Overall, this study may provide new insights that will improve our understanding of the roles of c-type lysozymes in the innate immunity of cyprinid fish, including the structural and phylogenetic characteristics, antimicrobial effects, and regulatory mechanism.


Assuntos
Anti-Infecciosos , Bactérias/metabolismo , Dexametasona/farmacologia , Proteínas de Peixes , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Carpa Dourada , Leptina/farmacologia , Lipopolissacarídeos/toxicidade , Muramidase , Transcrição Gênica/efeitos dos fármacos , Animais , Anti-Infecciosos/química , Anti-Infecciosos/metabolismo , Proteínas de Peixes/biossíntese , Proteínas de Peixes/química , Proteínas de Peixes/genética , Carpa Dourada/genética , Carpa Dourada/metabolismo , Muramidase/biossíntese , Muramidase/química , Muramidase/genética
17.
Gen Comp Endocrinol ; 283: 113240, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31394085

RESUMO

Leptin is primarily considered a peripheral satiety hormone and is also found to perform important roles in energy homeostasis in vertebrates ranging from fish to mammals. The liver is a major source of leptin production in teleost fish. Using goldfish as a model, a previous report by our group illustrated the positive regulation of leptin mRNA levels by treatment with the hyperglycemic hormone glucagon, and our present study provided evidence for the negative regulation of hepatic leptin-AI and leptin-AII transcripts through the administration of the hypoglycemic hormone insulin. This study is the first to demonstrate changes in the hepatopancreatic insulin, glucagon, leptin-AI and leptin-AII mRNA levels in goldfish during fasting and refeeding. Insulin was found to be effective in suppressing leptin-AI and leptin-AII transcript levels in goldfish liver via both in vivo intraperitoneal injection and in vitro cell incubation approaches. Only the insulin receptor, not the IGF-I receptor, was involved in insulin-inhibited leptin mRNA level. The suppression of leptin levels by insulin was caused by the activation of MKK3/6/p38MAPK and MEK1/2/Erk1/2 cascades. Insulin treatment could eliminate the stimulation of glucagon on leptin mRNA level. Our study describes the regulation and signal transduction mechanism of insulin on leptin mRNA levels in the goldfish liver, suggesting that the leptin function in fish is speculated to be not only an anorexigenic factor but also a metabolic mediator. This also supports the hypothesis that the poikilothermal fish use a passive survival strategy during the periods of food deprivation, which is mediated by the fish-specifically high leptin levels induced by the cooperation of insulin and glucagon.


Assuntos
Privação de Alimentos/fisiologia , Carpa Dourada/genética , Insulina/farmacologia , Leptina/genética , Fígado/metabolismo , Animais , Colforsina/farmacologia , Jejum , Comportamento Alimentar/efeitos dos fármacos , Glucagon/farmacologia , Hepatopâncreas/efeitos dos fármacos , Hepatopâncreas/metabolismo , Humanos , Injeções Intraperitoneais , Insulina/administração & dosagem , Fator de Crescimento Insulin-Like I/farmacologia , Fator de Crescimento Insulin-Like II/farmacologia , Leptina/metabolismo , Fígado/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores para Leptina/metabolismo , Transdução de Sinais/efeitos dos fármacos
18.
Nanotechnology ; 29(46): 465709, 2018 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-30188323

RESUMO

The optical confinement and strong carrier coupling within a semiconductor nanostructure cavity are crucial for the modulation of emission properties. Fundamental understanding of the light-matter interaction in a low dimensional system is important. In this paper, we synthesized high-quality hexagonal Te-doped CdS nanowires by two-step chemical vapor deposition and investigated systematically the doping concentration, temperature, excitation power, excitation wavelength dependent Raman, photoluminescence and carrier lifetime decay. Scanning electron microscopy, energy dispersive x-ray spectrometry and x-ray diffraction confirmed Te-doping in the as-prepared samples. The strong surface optical (SO) phonon mode is observed in the micro-Raman spectra of an individual Te-CdS nanowire, which is unsuitable in large-sized structures. In situ micro-photoluminescence (µ-PL) characterization shows dominant confined defect state emission with whispering gallery mode (WGM) characteristics. The emission peak position shifts under increased excitation power, demonstrating the inelastic scattering by bound carriers. In addition, the short wavelength emission modes are dominant at a low temperature (80 K) while the long wavelength emission modes are dominant at a high temperature (300 K) due to different recombination processes contributing to the WGM resonant bands, which was also confirmed by the time-resolved PL measurement. All these results reflect strong coupling between the surface evanescent-wave in the WGM cavity and the SO phonon/polaron, which will facilitate the rational tailoring of surface/interface relevant properties for nanophotonic device applications.

19.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 43(3): 229-239, 2018 Mar 28.
Artigo em Zh | MEDLINE | ID: mdl-29701183

RESUMO

OBJECTIVE: To explore the effect of intervention of E-cadherin (E-cad) and B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1) mediated by transcription activator-like effector nuclease (TALEN) on the biological behaviors of nasopharyngeal carcinoma cells.
 Methods: Multi-locus gene targeting vectors pUC-DS1-CMV-E-cad-2A-Neo-DS2 and pUC-DS1-Bmi-1 shRNA-Zeo-DS2 were constructed, and the E-cad and Bmi-1 targeting vectors were transferred with TALEN plasmids to CNE-2 cells individually or simultaneously. The integration of target genes were detected by PCR, the expressions of E-cad and Bmi-1 were detected by Western blot. The changes of cell proliferation were detected by cell counting kit-8 (CCK-8) assay. The cell cycle and apoptosis were detected by flow cytometry. The cell migration and invasion were detected by Transwell assay.
 Results: The E-cad and Bmi-1 shRNA expression elements were successfully integrated into the genome of CNE-2 cells, the protein expression level of E-cad was up-regulated, and the protein expression level of Bmi-1 was down-regulated. The intervention of E-cad and Bmi-1 didn't affect the proliferation, cell cycle and apoptosis of CNE-2 cells, but it significantly inhibited the migration and invasion ability of CNE-2 cells. Furthermore, the intervention of E-cad and Bmi-1 together significantly inhibited the migration ability of nasopharyngeal carcinoma cells compared with the intervention of E-cad or Bmi-1 alone (all P<0.01).
 Conclusion: The joint intervention of E-cad and Bmi-1 mediated by TALEN can effectively inhibit the migration and invasion of nasopharyngeal carcinoma cells in vitro, which may lay the preliminary experimental basis for gene therapy of human cancer.


Assuntos
Apoptose/fisiologia , Caderinas/fisiologia , Carcinoma/patologia , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Neoplasias Nasofaríngeas/patologia , Complexo Repressor Polycomb 1/fisiologia , Proteínas Proto-Oncogênicas/fisiologia , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição/metabolismo , Animais , Caderinas/genética , Carcinoma/genética , Carcinoma/metabolismo , Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Vetores Genéticos , Humanos , Técnicas In Vitro , Camundongos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/metabolismo , Complexo Repressor Polycomb 1/genética , Proteínas Proto-Oncogênicas/genética
20.
Int J Mol Sci ; 18(12)2017 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-29261147

RESUMO

Leptin actions at the pituitary level have been extensively investigated in mammalian species, but remain insufficiently characterized in lower vertebrates, especially in teleost fish. Prolactin (PRL) is a pituitary hormone of central importance to osmoregulation in fish. Using goldfish as a model, we examined the global and brain-pituitary distribution of a leptin receptor (lepR) and examined the relationship between expression of lepR and major pituitary hormones in different pituitary regions. The effects of recombinant goldfish leptin-AI and leptin-AII on PRL mRNA expression in the pituitary were further analysed, and the mechanisms underlying signal transduction for leptin-induced PRL expression were determined by pharmacological approaches. Our results showed that goldfish lepR is abundantly expressed in the brain-pituitary regions, with highly overlapping PRL transcripts within the pituitary. Recombinant goldfish leptin-AI and leptin-AII proteins could stimulate PRL mRNA expression in dose- and time-dependent manners in the goldfish pituitary, by both intraperitoneal injection and primary cell incubation approaches. Moreover, the PI3K/Akt/mTOR, MKK3/6/p38MAPK, and MEK1/2/ERK1/2-but not JAK2/STAT 1, 3 and 5 cascades-were involved in leptin-induced PRL mRNA expression in the goldfish pituitary.


Assuntos
Proteínas de Peixes/metabolismo , Leptina/farmacologia , Sistema de Sinalização das MAP Quinases , Hipófise/metabolismo , Prolactina/metabolismo , Animais , Células Cultivadas , Proteínas de Peixes/genética , Carpa Dourada , Fosfatidilinositol 3-Quinases/metabolismo , Hipófise/efeitos dos fármacos , Prolactina/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores para Leptina/genética , Receptores para Leptina/metabolismo , Serina-Treonina Quinases TOR/metabolismo
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