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1.
BMC Musculoskelet Disord ; 25(1): 15, 2024 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-38166826

RESUMO

BACKGROUND: The majority of published literature clinically assesses surgical outcomes after lower limb replantation for traumatic amputations. However, patients' satisfaction and quality of life may not be accurately measured through rigid scoring using standardized patient reported outcome measures. PURPOSE: The aim of this study was to qualitatively assess patient satisfaction and factors associated with achieving good outcomes after successful lower limb replantation surgery. METHODS: A semi-structured interview was conducted with 12 patients who underwent lower limb replantation surgery following traumatic amputation injuries. The interview focused on the patients' experience and satisfaction throughout their injury, surgical journey, rehabilitation and reintegration into their communities. An inductive and deductive thematic analysis was applied using the recorded transcripts to evaluate the overall satisfaction of the patients after lower limb replantation surgery. RESULTS: The following observations emerged from the structured themes among all the patients interviewed: (1) Family and social support was significantly associated with improved qualities of life and satisfaction after lower limb replantation; (2) Patients were generally satisfied with their outcomes despite limitations in physical capabilities; (3) Satisfaction was associated with acceptance of their cosmetic deformity; (4) Social integration and being able to participate in a meaningful manner was associated with greater satisfaction after recovery. CONCLUSIONS: Patients who undergo lower limb replantation can have a significantly improved quality of life if they have strong social support, are able to contribute in a meaningful manner to their communities after surgery, and are accepting of their cosmetic deficiencies.


Assuntos
Amputação Traumática , Satisfação do Paciente , Humanos , Qualidade de Vida , Reimplante , Amputação Traumática/cirurgia , Extremidade Inferior/cirurgia
2.
Bioorg Chem ; 138: 106637, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37276679

RESUMO

Treatment of breast cancer has greatly evolved during the last decades, but triple negative breast cancer (TNBC) with a higher degree of malignancy cannot be directly and effectively treated. Abnormal cell cycle is generally found in human breast cancer and other malignant tumors, and cyclin-dependent kinases (CDK) 4/6, a cell cycle-related regulatory nuclear protein, is deemed as an effective target for breast cancer treatment so far. Since DCAF16 E3 ligase is also mainly distributed in the nucleus, in this study, by combining Palbociclib and DCAF16 E3 ligase ligand KB02 with different linkers, a series of DCAF16 based CDK4/6 degraders were designed and synthesized. Among them, compound A4 showed potent inhibitory activity against CDK4/6, and decreased the level of CDK4/6 protein in MDA-MB-231 cells in a concentration- and time-dependent manner. Moreover, the toxicity of A4 in normal cells showed 7 times lower than that of Palbociclib, and A4 exhibits therapeutic potential in MDA-MB-231 xenograft models in vivo. These findings indicate that A4, as a novel CDK4/6 degrader based on DCAF16, is worthy of further investigating for the treatment of TNBC.


Assuntos
Neoplasias de Mama Triplo Negativas , Humanos , Neoplasias de Mama Triplo Negativas/metabolismo , Proliferação de Células , Linhagem Celular Tumoral , Ciclo Celular , Divisão Celular , Quinase 4 Dependente de Ciclina
3.
Cryobiology ; 92: 235-240, 2020 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-32001216

RESUMO

Every year tens of millions reconstruction surgeries were performed to repair tissue defects caused by trauma or malignant tumor. As the composite tissues are very difficult to be preserved for a long time in vitro, "organ bank" was proposed. In addition to immune rejection, tissue cryopreservation is the key step of the "Organ Bank". In this study, two severed fingers were cryopreserved because the patients' conditions were not allowed to undergo the replantation immediately. The fingers were perfused with the cryopreservation solution through the digital artery at a speed of 1 ml/min, they were frozen following a control rate freezing procedure and stored in the liquid nitrogen. One finger was cryopreserved for 10 days and the other for 30 days. Both fingers were successfully replanted when the patient get better. After 1 year follow-up, the replanted fingers achieved satisfactory appearance and function. Our result demonstrate small composite tissues such as severed fingers can be cryopreserved and replanted successfully, which will help the patients who cannot receive the replantation immediately. This method will have important application value in cryobiology and clinical medicine.


Assuntos
Criopreservação/métodos , Dedos/cirurgia , Procedimentos de Cirurgia Plástica/métodos , Reimplante/métodos , Adulto , Criobiologia , Congelamento , Humanos , Masculino , Pessoa de Meia-Idade , Ferimentos e Lesões/cirurgia
4.
J Cell Biochem ; 120(10): 18162-18171, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31144385

RESUMO

BACKGROUND: This study was to investigate the effects of recombinant human growth hormone (rhGh) on ischemia-reperfusion (I/R) injury of mouse flaps. METHODS: Healthy mice were randomly divided into four groups as follows: sham group, the IR group, the sham+rhGH group and the IR+rhGH group, with 12 mice in each group. Skin pathology was tested by hematoxylin and eosin staining. The flap survival of each group was measured after 7 days. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were determined using corresponding kit. The levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α in serum and flap were respectively measured by performing enzyme-linked immunosorbent assay and quantitative real-time (qRT)-PCR. The expressions of cleaved caspase-3, B cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated X protein (Bax), vascular endothelial growth factor (VEGF), MnSOD, toll-like receptor 4 (TLR4), and galectin-3 and (p)-p65 were analyzed by RT-PCR or/and Western blot. RESULTS: Prophylactic systemic application of recombinant human GH reduced the pathological damage of skin IR and significantly improved the flap survival of IR in mice, accompanied by elevation of VEGF. After administration of recombinant human GH, the activity of SOD/MnSOD in the flap was significantly increased, while the content of MDA was decreased. Cleaved caspase-3 and Bax were downregulated and Bcl-2 was upregulated in IR+rhGH group, compared to IR group. The levels of TLR4, Galectin-3 and p-p65 were decreased by rhGH. CONCLUSION: rhGH had protective effects on flap IR injury, and can be used as a drug intervention target for the treatment of skin flap IR injury.


Assuntos
Apoptose/efeitos dos fármacos , Hormônio do Crescimento Humano/farmacologia , Inflamação/prevenção & controle , Traumatismo por Reperfusão/fisiopatologia , Retalhos Cirúrgicos/fisiologia , Animais , Apoptose/fisiologia , Caspase 3/genética , Caspase 3/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Inflamação/metabolismo , Inflamação/fisiopatologia , Interleucina-6/sangue , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Superóxido Dismutase/metabolismo , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
5.
Sci Rep ; 14(1): 6341, 2024 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-38491143

RESUMO

The conservative treatment for Kohler's disease will take several months, but some patients still have flatfoot and persistent pain. From October 2013 to July 2015, 3 children with Kohler's disease underwent navicular decompression and micro-circulation reconstruction surgery in our hospital. All the patients have received conservative treatment for more than 3 months and the effect was poor. X-ray showed the bone density of navicular increased significantly. All patients were followed up over 1 year. The 3 patients recovered well. VAS score decreased from 7.0 to 2.6 at 1 month after the operation. The pain symptom disappeared completely on 3 months after surgery. The density of navicular bone recovered to normal. Navicular decompression and micro-circulation reconstruction surgery may quickly improve the ischemic status of navicular bone, alleviate pain symptom and enable patients to resume normal activity as soon as possible.


Assuntos
Pé Chato , Osteocondrite , Osteocondrose , Ossos do Tarso , Criança , Humanos , Dor/etiologia , Dor/cirurgia
6.
Aging (Albany NY) ; 15(21): 11918-11939, 2023 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-37921852

RESUMO

Cutaneous melanoma (CM) is widely acknowledged as a highly aggressive form of malignancy that is associated with a considerable degree of morbidity and poor prognosis. Despite this recognition, the precise role of hypoxia-related long noncoding RNAs (HRLs) in the pathogenesis of CM remains an area of active research. This study sought to elucidate the contribution of HRLs in CM by conducting a thorough screening and extraction of hypoxia-related genes (HRGs). In particular, we conducted univariate and multivariate Cox regression analyses to assess the independence of the prognostic signature of HRLs. Our results demonstrated that a novel risk model could be established based on five prognostic HRLs. Remarkably, patients with low-risk scores exhibited significantly higher overall survival rates compared to their high-risk counterparts, as confirmed by Kaplan-Meier survival analysis. Furthermore, we utilized consensus clustering analysis to categorize CM patients into two distinct subtypes, which revealed marked differences in their prognosis and immune infiltration landscapes. Our nomogram results confirmed that the HRLs prognostic signature served as an independent prognostic indicator, offering an accurate evaluation of the survival probability of CM patients. Notably, our findings from ESTIMATE and ssGSEA analyses highlighted significant disparities in the immune infiltration landscape between low- and high-risk groups of CM patients. Additionally, IPS and TIDE results suggested that CM patients in different risk subtypes may exhibit favorable responses to immunotherapy. Enrichment analysis and GSVA results indicated that immune-related signaling pathways may mediate the role of HRLs in CM. Finally, our tumor mutation burden (TMB) results indicated that patients with low-risk scores had a higher TMB status. In summary, the establishment of a risk model based on HRLs in this study provided an accurate prognostic prediction and correlated with the immune infiltration landscape of CM, thereby providing novel insights for the future clinical management of this disease.


Assuntos
Melanoma , Neoplasias Cutâneas , Humanos , Melanoma/genética , Melanoma/terapia , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/terapia , Prognóstico , Hipóxia , Imunoterapia , Microambiente Tumoral , Melanoma Maligno Cutâneo
7.
J Pers Med ; 13(1)2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36675804

RESUMO

Reconstruction surgeries in the inguinal area are challenging for vascular surgeons, oncologists, orthopedists, and others. The pedicled sartorius flap is the most commonly used flap for reconstruction. The pedicled adductor longus is reported as a new method to reconstruct the inguinal region. The related anatomic study is rare. This work aims to make a comparison of pedicled adductor longus and pedicled sartorius on cadavers for better use. Out of the 12 thighs in the 6 cadavers analyzed, the author compares two surgical methods in terms of surgical details, exposure of vascular and nerve pedicle, flap harvesting, flap transposition and flap volume, etc. Through the course of this study, it is showed that the adductor longus flap had a sizable advantage over the sartorius flap in terms of exposure, harvesting, and flap volume.

8.
Comput Assist Surg (Abingdon) ; 28(1): 2198106, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37070416

RESUMO

BACKGROUND: The treatment of talus avascular necrosis (AVN) is challenging owing to its unique anatomical features. Despite decades of studies, till date, there is no appropriate treatment for talus AVN. Therefore, surgeons need to develop newer surgical methods. In the present study we introduce a new surgical method, 3D printed partial talus replacement (PTR), to treat partial talus necrosis and collapse (TNC). METHODS: A male patient with talus AVN underwent PTR in our hospital. The morphology of the talus was quantified using 3D computed tomography (CT) imaging. A novel 3D printed titanium prothesis was designed and manufactured according to the findings of the CT imaging. The prosthesis was applied during talus replantation surgery to reconstruct the anatomical structure of the ankle. The follow-up period for this patient was 24 months. The visual analog scale (VAS) scores before and after surgery, American Orthopedic Foot and Ankle Score (AOFAS), ankle range of motion, and postoperative complications were recorded to evaluate the prognosis. RESULTS: The anatomical structure of the talus was reconstructed. The patient was satisfied with the effects of treatment, recovery, and function. The VAS score decreased from 5 to 1. The AOFAS improved from 70 to 93. The range of motion remained the same as that during the pre-operation. The patient returned to a normal life. CONCLUSION: 3D printed PTR is a new surgical method for talus AVN that can provide satisfactory outcomes. In future, PTR might be an effective and preferential treatment for the treatment of partial talus AVN and collapse.


Assuntos
Impressão Tridimensional , Próteses e Implantes , Tálus , Humanos , Masculino , Necrose , Reimplante , Estudos Retrospectivos , Tálus/cirurgia , Tálus/transplante
9.
Mol Biomed ; 3(1): 17, 2022 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-35654983

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the coronavirus disease 2019 (COVID-19) global pandemic. Omicron, a new variant of SARS-CoV-2, has the characteristics of strong transmission and pathogenicity, short incubation period, and rapid onset progression, and has spread rapidly around the world. The high replication rate and intracellular accumulation of SARS-CoV-2 are remarkable, but the underlying molecular mechanisms remain unclear. Autophagy acts as a conservative cellular defence mechanism against invading pathogens. Here, we provide evidence that the main protease of SARS-CoV-2, NSP5, effectively cleaves the selective autophagy receptor p62. NSP5 targets p62 for cleavage at glutamic acid 354 and thus abolishes the capacity of p62 to mediate selective autophagy. It was further shown that p62 specifically interacted with ubiquitinated SARS-CoV-2 M, the viral membrane protein, to promote its autophagic degradation. In the presence of NSP5, p62-mediated autophagic degradation of the M protein was inhibited. The cleaved products of p62 also cannot facilitate the degradation of the M protein. Collectively, our findings reveal that p62 is a novel host target of SARS-CoV-2 NSP5 and suggest that selective autophagy targets viruses and potential strategies by which the virus evades autophagic clearance. Our results may provide new ideas for the development of anti-COVID-19 drugs based on autophagy and NSP5.

10.
J Plast Reconstr Aesthet Surg ; 75(11): 4042-4047, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36207234

RESUMO

BACKGROUND: Foot injuries due to vehicular or other accidents are common. However, complete toe amputation is rare. This study explored the current protocols and clinical significance of toe replantation. METHODS: From December 2011 to December 2018, ten patients with 13 severed toes underwent toe replantation in our hospital. Seven cases were replanted antegrade, and three cases were replanted retrograde. RESULTS: All patients were followed for two to three years after toe replantation. One big toe underwent necrosis, while the other 12 toes survived completely. The appearance and feel of the successfully replanted toes were satisfactory, and the patients exhibited a normal gait. CONCLUSION: Toe replantation can achieve an acceptable appearance and function of the foot and considerably reduce the psychological effects experienced by the patients. Increased clinical attention and application of toe replantation are needed. LEVEL OF EVIDENCE: Level IV, retrospective case series.


Assuntos
Amputação Traumática , Traumatismos dos Dedos , Hallux , Humanos , Amputação Traumática/cirurgia , Estudos Retrospectivos , Reimplante/métodos , Dedos do Pé/cirurgia , Dedos do Pé/lesões , Microcirurgia/métodos , Traumatismos dos Dedos/cirurgia
11.
J Plast Reconstr Aesthet Surg ; 75(7): 2387-2440, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35508521

RESUMO

Degloving injury is a common and intractable injury with the bone and tendon exposed and contamination, the stripped skin cannot be replanted immediately and will be discarded, although auto-graft is needed for subsequent wound repair. In this study, autologous skin cryopreservation technique was applied to the treatment of severe limb degloving injuries. The clinical data of 9 patients from January 2016 to December 2018 were analyzed retrospectively. Among the 9 cases, 1 case developed necrosis due to wound infection, and the rest survived 60-100%. The replanted cryopreserved skin were soft and resilient, with poor sensory recovery, varying degrees of discoloration and no hair growth. Cryopreservation provides more time for improving the wound and whole-body condition. The frozen skin had good quality and high survival rate. Our study can effectively use the degloving skin, reduce the damage of the donor area.


Assuntos
Avulsões Cutâneas , Procedimentos de Cirurgia Plástica , Criopreservação , Avulsões Cutâneas/cirurgia , Humanos , Procedimentos de Cirurgia Plástica/métodos , Reimplante/métodos , Estudos Retrospectivos , Transplante de Pele/métodos , Resultado do Tratamento
12.
Comput Assist Surg (Abingdon) ; 26(1): 15-21, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-33625935

RESUMO

BACKGROUND: Charcot neuroarthropathy of the ankle is an extremely challenging clinical dilemma, and its surgical management can be highly complicated. The goal of Charcot ankle treatment is to to restore a plantigrade and stable foot, and thus to avoid ulceration with subsequent infection. This report aims to introduce a method of correcting ankle deformity using a novel 3D printing technique. PATIENT AND METHODS: One patient with Charcot ankle deformity was operated in this study. The ankle deformity of this patient was quantified in three dimensions through computed tomography. On the basis of the computed tomography scans, a new titanium implant was designed and manufactured using 3D printing. The implant was applied in the surgery of tibio-talo-calcaneal arthrodesis to restore local anatomy of the affected ankle of the patient with Charcot neuroarthropathy. RESULTS: Evaluation of the post-operative radiography indicated union in the affected ankle. After surgery, the planar foot in this patient was restored. The patient was satisfied with the post-operative course, and joint fusion was successful as indicated by 2-year post-operative evaluation. The results of post-operative follow-up showed that the lower limb length of the patient with Charcot neuroarthropathy was salvaged, and the patient retained the plantigrade foot. CONCLUSION: Three-dimensional printing technique combined with tibio-talo-calcaneal arthrodesis may help to correct ankle deformity in patients with Charcot neuroarthropathy.


Assuntos
Tornozelo , Titânio , Articulação do Tornozelo/diagnóstico por imagem , Articulação do Tornozelo/cirurgia , Artrodese , Humanos , Próteses e Implantes
13.
Cell Rep ; 37(7): 109937, 2021 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-34788606

RESUMO

Acetyl ligation to the amino acids in a protein is an important posttranslational modification. However, in contrast to lysine acetylation, N-terminal acetylation is elusive in terms of its cellular functions. Here, we identify Nat3 as an N-terminal acetyltransferase essential for autophagy, a catabolic pathway for bulk transport and degradation of cytoplasmic components. We identify the actin cytoskeleton constituent Act1 and dynamin-like GTPase Vps1 (vacuolar protein sorting 1) as substrates for Nat3-mediated N-terminal acetylation of the first methionine. Acetylated Act1 forms actin filaments and therefore promotes the transport of Atg9 vesicles for autophagosome formation; acetylated Vps1 recruits and facilitates bundling of the SNARE (soluble N-ethylmaleimide-sensitive factor activating protein receptor) complex for autophagosome fusion with vacuoles. Abolishment of the N-terminal acetylation of Act1 and Vps1 is associated with blockage of upstream and downstream steps of the autophagy process. Therefore, our work shows that protein N-terminal acetylation plays a critical role in controlling autophagy by fine-tuning multiple steps in the process.


Assuntos
Autofagia/fisiologia , Acetiltransferase N-Terminal B/metabolismo , Acetiltransferases N-Terminal/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Acetilação , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Autofagossomos/metabolismo , Autofagia/genética , Proteínas de Transporte/metabolismo , China , Proteínas de Ligação ao GTP/metabolismo , Acetiltransferase N-Terminal B/fisiologia , Acetiltransferases N-Terminal/fisiologia , Fagossomos/metabolismo , Processamento de Proteína Pós-Traducional , Transporte Proteico/fisiologia , Proteínas SNARE/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/fisiologia , Vacúolos/metabolismo , Proteínas de Transporte Vesicular/metabolismo
14.
Autophagy ; 17(12): 4512-4514, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34705610

RESUMO

Macroautophagy/autophagy is special because the double-layer lipid-formed autophagosome is formed by de novo generation. Phosphatidylinositol-3-phosphate (PtdIns3P) produced by class III phosphatidylinositol 3-kinase complex I (PtdIns3K-CI) is an essential source lipid for the formation of autophagosomes. However, how autophagy is initiated is unknown. In other words, the mechanism by which PtdIns3K-CI is recruited to the phagophore assembly site (PAS) to initiate autophagosome formation is unclear. We recently uncovered the pivotal role of yeast Vac8 in autophagy initiation through the recruitment of PtdIns3K-CI to the PAS. N-terminal palmitoylation of Vac8 anchors it to the vacuole membrane, and the middle ARM domains bind PtdIns3K-CI, leading to the generation of PtdIns3P at the PAS and subsequent autophagosome formation. We found that mouse ARMC3 is the homolog of yeast Vac8 and that its autophagic roles are conserved. Interestingly, spermatids from mice with Armc3 deletion showed blocked ribophagy, low energy levels of mitochondria and motionless flagella, which caused male infertility. These findings revealed a germ tissue-specific autophagic function of ARMC3 in complex eukaryotic species.


Assuntos
Proteínas do Domínio Armadillo , Autofagia , Classe III de Fosfatidilinositol 3-Quinases , Proteínas de Saccharomyces cerevisiae , Animais , Proteínas do Domínio Armadillo/metabolismo , Autofagossomos/metabolismo , Autofagia/fisiologia , Classe III de Fosfatidilinositol 3-Quinases/metabolismo , Masculino , Camundongos , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Transdução de Sinais , Espermatogênese , Proteínas de Transporte Vesicular/metabolismo
15.
Autophagy ; 17(8): 1907-1917, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-32693712

RESUMO

Biogenesis of autophagosomes is the early step of macroautophagy/autophagy and requires membrane acquisition mainly from ER-Golgi-sourced precursor vesicles. Matured autophagosomes fuse with lysosomes for final degradation. However, how this selective fusion is determined remains elusive. Here, we identified Sac1 by a high throughput screen in Saccharomyces cerevisiae to show it was critical for autophagosome-lysosome fusion through its PtdIns4P phosphatase activity. Sac1 deficiency caused a dramatic increase of PtdIns4P at early Golgi apparatus and abnormal incorporation of PtdIns4P into Atg9 vesicles and autophagosomes, which caused failure to recruit SNARE proteins for autophagosome fusion with vacuoles. Sac1 function in autophagy was highly conserved from yeast to mammalian cells. Our work thus suggested that correct upstream lipid incorporation was important for downstream fusion step of autophagy and that Sac1 played a critical and ancient role in this surveillance of lipid integration.Abbreviations: Ape1: aminopeptidase Ι; ATG: autophagy related; EBSS: Earle's balanced salt solution; ER: endoplasmic reticulum; ERGIC: Golgi apparatus and ER-Golgi intermediate compartment; HOPS: homotypic fusion and protein sorting complex; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; PtdIns3P: phosphatidylinositol-3-phosphate; PtdIns4K: phosphoinositide-4-kinase; PtdIns4P: phosphatidylinositol-4-phosphate; SD-N: nitrogen starvation medium; SNARE: soluble N-ethylamide-sensitive factor attachment protein receptor.


Assuntos
Autofagossomos/metabolismo , Autofagia/fisiologia , Fosfatos de Fosfatidilinositol/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Retículo Endoplasmático/metabolismo , Complexo de Golgi/metabolismo , Células HeLa , Humanos , Hidrolases/metabolismo , Lisossomos/metabolismo , Fusão de Membrana , Saccharomyces cerevisiae/metabolismo
16.
Dev Cell ; 56(16): 2313-2328.e7, 2021 08 23.
Artigo em Inglês | MEDLINE | ID: mdl-34428398

RESUMO

How autophagy initiation is regulated and what the functional significance of this regulation is are unknown. Here, we characterized the role of yeast Vac8 in autophagy initiation through recruitment of PIK3C3-C1 to the phagophore assembly site (PAS). This recruitment is dependent on the palmitoylation of Vac8 and on its middle ARM domains for binding PIK3C3-C1. Vac8-mediated anchoring of PIK3C3-C1 promotes PtdIns3P generation at the PAS and recruitment of the PtdIns3P binding protein Atg18-Atg2. The mouse homolog of Vac8, ARMC3, is conserved and functions in autophagy in mouse testes. Mice lacking ARMC3 have normal viability but show complete male infertility. Proteomic analysis indicated that the autophagic degradation of cytosolic ribosomes was blocked in ARMC3-deficient spermatids, which caused low energy levels of mitochondria and motionless flagella. These studies uncovered a function of Vac8/ARMC3 in PtdIns3-kinase anchoring at the PAS and its physical significance in mammalian spermatogenesis with a germ tissue-specific autophagic function.


Assuntos
Autofagia , Ribossomos/metabolismo , Cauda do Espermatozoide/metabolismo , Espermatogênese , Adulto , Animais , Autofagossomos/metabolismo , Células Cultivadas , Classe III de Fosfatidilinositol 3-Quinases/metabolismo , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Saccharomyces cerevisiae , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Motilidade dos Espermatozoides , Cauda do Espermatozoide/fisiologia , Proteínas de Transporte Vesicular/genética , Proteínas de Transporte Vesicular/metabolismo
17.
J Invest Surg ; 33(5): 422-427, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30884993

RESUMO

Purpose: With increasing use of the chimeric flap of the descending genicular artery, the authors systematically investigated the anatomy of its branches in cadavers. Methods: Fifteen fresh cadaveric thighs were studied by anatomical dissection. The branches of the descending genicular arteries were skeletonized along their courses to the femoral arteries. Branches' lengths and diameters were measured to simulate the combined application of the skin, muscle, bone, osteochondral and osteocutaneous flaps with tendon enthesis. Results: The descending genicular artery was noted in 11 thighs, with an average diameter of 1.94 ± 0.36 mm and an average length of 10.69 ± 4.41 mm. In addition, the saphenous artery was noted in all 15 thighs, and the average diameter of the original part was 1.35 ± 0.18 mm. Branches arose from the saphenous artery to supply the skin above the knee, the anterior of tibia, the sartorius muscle and the pes anserinus. The average diameter of the osteoarticular artery was 1.80 ± 0.46 mm which divaricated into a periosteal branch to supply the bone above the medial femoral epicondyle and a few articular branches to supply the bone and the cartilage of the medial femoral condyle. Conclusions: This study systematically investigated the anatomy of the descending genicular artery and its branches. Based on the anatomical features of descending genicular artery, chimeric flap offers combination therapy with other tissue flaps. Besides, considering its long chimeric arm, chimeric flap could be used to repair not only local complex injuries but also defects in different locations. Clinical Relevance: The descending genicular artery chimeric flap is a clinical option for reconstructing compound tissue defects of limbs.


Assuntos
Artéria Femoral/anatomia & histologia , Joelho/irrigação sanguínea , Retalho Perfurante/irrigação sanguínea , Cadáver , Terapia Combinada/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Retalho Perfurante/transplante , Procedimentos de Cirurgia Plástica/métodos
18.
Sci Rep ; 9(1): 16171, 2019 11 07.
Artigo em Inglês | MEDLINE | ID: mdl-31700003

RESUMO

Osteoblasts are implicated in the building of the vertebrate skeleton. The current study aimed to investigate the role of microRNA-495 (miR-495) in the osteoblasts of mice with tibial fractures and the underlying mechanism involving in aquaporin-1 (AQP1) and the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. Initially, a microarray-based analysis was performed to screen the differentially expressed genes and miRNAs associated with tibial fracture. Following the establishment of a tibial fracture mouse model, the positive rate of the AQP1 protein in the fracture tissue was detected by immunohistochemistry (IHC). Next, to verify the binding site between miR-495 on AQP1, bioinformatics data were employed in addition to the application of a dual-luciferase reporter gene assay. The osteoblast cell line MC3T3-E1 was treated with miR-495 mimic, miR-495 inhibitor and Anisomycin to explore the potent effects of miR-495 on proliferation and differentiation of osteoblasts in mice with tibial fracture. The expression of miR-495, AQP1, p38 MAPK, PCNA, Cyclin D1, OCN, and OPN was subsequently evaluated by RT-qPCR and Western blot analysis. Cell viability, the number of calcium nodules and alkaline phosphatase (ALP) activity were detected by MTT assay, alizarin red staining, and ALP activity assay, respectively. Our results revealed that miR-495 was down-regulated while AQP1 was up-regulated in the mice with tibial fractures. AQP1 was verified as a target gene of miR-495. When the cells were treated with overexpressed miR-495 or activated p38 MAPK signaling pathway, elevated expression of PCNA, Cyclin, D1, OCN, and OPN along with an increased amount of calcium nodules, higher cell viability, and enhanced ALP activity was detected, while the expression of AQP1 was reduced. Collectively, the key findings of the present study support the notion that overexpressed miR-495 may activate the p38 MAPK signaling pathway to inhibit AQP1 and to promote the proliferation and differentiation of osteoblasts in mice with tibial fracture.


Assuntos
Aquaporina 1/biossíntese , Diferenciação Celular , Proliferação de Células , Regulação para Baixo , Sistema de Sinalização das MAP Quinases , MicroRNAs/metabolismo , Osteoblastos/metabolismo , Fraturas da Tíbia/metabolismo , Animais , Linhagem Celular , Camundongos , Osteoblastos/patologia , Fraturas da Tíbia/patologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
19.
Indian J Orthop ; 48(6): 594-8, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25404772

RESUMO

BACKGROUND: Fingertip defect can be treated with many flaps such as random pattern abdominal flap, retrograde digital artery island flap, V-Y advancement flap, etc. However, swelling in the fingertip, dysfunction of sensation, flexion and extension contracture or injury in the hemi-artery of the finger usually occurs during the recovery phase. Recently, digital artery perforator flaps have been used for fingertip reconstructions. With the development of super microsurgery techniques, free flaps can be more effective for sensory recovery and durability of the fingertip. MATERIALS AND METHODS: Six cases (six fingers) of fingertip defects were treated with free digital artery perforator flaps of appropriate size and shape from the proximal phalanx. During surgery, the superficial veins at the edge of flap were used as reflux vessels and the branches of the intrinsic nerve and dorsal digital nerve toward the flap were used as sensory nerves. The proximal segment of the digital artery (cutaneous branches) towards the flap was cut off to form the pedicled free flap. The fingertips were reconstructed with the free flap by anastomosing the cutaneous branches of digital artery in the flap with the distal branch or trunk of the digital artery, the flap nerve with the nerve stump and the veins of the flap with the digital artery accompanying veins or the superficial veins in the recipient site. RESULTS: Six flaps survived with successful skin grafting. Patients were followed up for 6-9 months. The appearance and texture of the flaps was satisfactory. The feeling within the six fingers recovered to S4 level (BMRC scale) and the two point discrimination was 3-8 mm. CONCLUSION: Free digital artery perforator flap is suitable for repairing fingertip defect, with good texture, fine fingertip sensation and without sacrificing the branch of the digital artery or nerve.

20.
Artigo em Chinês | MEDLINE | ID: mdl-17190419

RESUMO

OBJECTIVE: To report replantation of amputated ear with microtechnique in 5 cases. METHODS: From 2002 to 2005, 5 totally amputated ears were replanted and 4 obediently, 1 retrogressively. In every case, 1-2 arteries and 1-3 veins were anastomosed. Seven vessels were detective and 1-6 cm vessels were transplanted from forearm or dorsal side of hand. Amputated ears obtained blood supply again in about 6-10 hours after injury and exploration was enforced for venous crisis in 2 cases, and bloodletting in 1 case. RESULTS: All 5 cases survived. Auricles possess perfect shape, no pigmentation, slight atrophy and perfect sensation. CONCLUSIONS: Amputated ear should be replanted as long as no obvious contusion occurred, and the keys to prevent vascular crisis are cutting off unhealthy vessels, grafting superficial veins for bridging, and the high quality of vascular anastomosis.


Assuntos
Anastomose Cirúrgica/métodos , Orelha Externa/cirurgia , Microvasos/cirurgia , Reimplante/métodos , Adulto , Orelha Externa/lesões , Feminino , Humanos , Masculino , Microcirurgia , Adulto Jovem
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