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1.
Cell ; 182(6): 1531-1544.e15, 2020 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-32846158

RESUMO

The fidelity of intracellular signaling hinges on the organization of dynamic activity architectures. Spatial compartmentation was first proposed over 30 years ago to explain how diverse G protein-coupled receptors achieve specificity despite converging on a ubiquitous messenger, cyclic adenosine monophosphate (cAMP). However, the mechanisms responsible for spatially constraining this diffusible messenger remain elusive. Here, we reveal that the type I regulatory subunit of cAMP-dependent protein kinase (PKA), RIα, undergoes liquid-liquid phase separation (LLPS) as a function of cAMP signaling to form biomolecular condensates enriched in cAMP and PKA activity, critical for effective cAMP compartmentation. We further show that a PKA fusion oncoprotein associated with an atypical liver cancer potently blocks RIα LLPS and induces aberrant cAMP signaling. Loss of RIα LLPS in normal cells increases cell proliferation and induces cell transformation. Our work reveals LLPS as a principal organizer of signaling compartments and highlights the pathological consequences of dysregulating this activity architecture.


Assuntos
Carcinogênese/metabolismo , Carcinoma Hepatocelular/genética , Compartimento Celular/genética , Subunidade RIalfa da Proteína Quinase Dependente de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Proteínas de Choque Térmico HSP40/genética , Neoplasias Hepáticas/genética , Transdução de Sinais , Animais , Carcinogênese/efeitos dos fármacos , Carcinogênese/genética , Carcinoma Hepatocelular/metabolismo , Compartimento Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , AMP Cíclico/farmacologia , Subunidade RIalfa da Proteína Quinase Dependente de AMP Cíclico/genética , Proteínas Quinases Dependentes de AMP Cíclico/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Citoplasma/metabolismo , Humanos , Neoplasias Hepáticas/metabolismo , Camundongos , Oncogenes/genética , Domínios Proteicos , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes de Fusão , Espectroscopia de Infravermelho com Transformada de Fourier , Imagem com Lapso de Tempo/métodos
2.
Nat Chem Biol ; 20(2): 151-161, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37500769

RESUMO

G-protein-coupled receptors (GPCRs) can initiate unique functional responses depending on the subcellular site of activation. Efforts to uncover the mechanistic basis of compartmentalized GPCR signaling have concentrated on the biochemical aspect of this regulation. Here we assess the biophysical positioning of receptor-containing endosomes as an alternative salient mechanism. We devise a strategy to rapidly and selectively redistribute receptor-containing endosomes 'on command' in intact cells without perturbing their biochemical composition. Next, we present two complementary optical readouts that enable robust measurements of bulk- and gene-specific GPCR/cyclic AMP (cAMP)-dependent transcriptional signaling with single-cell resolution. With these, we establish that disruption of native endosome positioning inhibits the initiation of the endosome-dependent transcriptional responses. Finally, we demonstrate a prominent mechanistic role of PDE-mediated cAMP hydrolysis and local protein kinase A activity in this process. Our study, therefore, illuminates a new mechanism regulating GPCR function by identifying endosome positioning as the principal mediator of spatially selective receptor signaling.


Assuntos
Endossomos , Transdução de Sinais , Transdução de Sinais/fisiologia , Endossomos/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , AMP Cíclico/metabolismo , Fosforilação
3.
Nat Chem Biol ; 2024 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-39394268

RESUMO

The protein kinase C (PKC) family of serine and threonine kinases, consisting of three distinctly regulated subfamilies, has been established as critical for various cellular functions. However, how PKC enzymes are regulated at different subcellular locations, particularly at emerging signaling hubs, is unclear. Here we present a sensitive excitation ratiometric C kinase activity reporter (ExRai-CKAR2) that enables the detection of minute changes (equivalent to 0.2% of maximum stimulation) in subcellular PKC activity. Using ExRai-CKAR2 with an enhanced diacylglycerol (DAG) biosensor, we uncover that G-protein-coupled receptor stimulation triggers sustained PKC activity at the endoplasmic reticulum and lysosomes, differentially mediated by Ca2+-sensitive conventional PKC and DAG-sensitive novel PKC, respectively. The high sensitivity of ExRai-CKAR2, targeted to either the cytosol or partitioning defective complexes, further enabled us to detect previously inaccessible endogenous atypical PKC activity in three-dimensional organoids. Taken together, ExRai-CKAR2 is a powerful tool for interrogating PKC regulation in response to physiological stimuli.

4.
Annu Rev Pharmacol Toxicol ; 61: 587-608, 2021 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-33411579

RESUMO

How cells muster a network of interlinking signaling pathways to faithfully convert diverse external cues to specific functional outcomes remains a central question in biology. Through their ability to convert dynamic biochemical activities to rapid and precise optical readouts, genetically encoded fluorescent biosensors have become instrumental in unraveling the molecular logic controlling the specificity of intracellular signaling. In this review, we discuss how the use of genetically encoded fluorescent biosensors to visualize dynamic signaling events within their native cellular context is elucidating the different strategies employed by cells to organize signaling activities into discrete compartments, or signaling microdomains, to ensure functional specificity.


Assuntos
Técnicas Biossensoriais , Transferência Ressonante de Energia de Fluorescência , Humanos , Transdução de Sinais
5.
Apoptosis ; 29(3-4): 344-356, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-37848674

RESUMO

BACKGROUND: Pyroptosis, as a type of inflammatory programmed cell death, has been studied in inflammatory diseases and numerous cancers but its role in pancreatic ductal adenocarcinoma (PDAC) remains further exploration. METHODS: A TCGA-PDAC cohort was enrolled for bioinformatics analysis to investigate the effect of pyroptosis on the prognosis and drug sensitivity of patients. PA-TU-8988T and CFPAC-1 cells were selected for investigating the role of GSDMC in PDAC. RESULTS: A distinct classification pattern of PDAC mediated by 21 pyroptosis-related genes (PRGs) was identified. It was suggested that higher pyroptosis activity was associated with poor prognosis of patients and higher tumor proliferation rates. We further established a prognostic model based on three PRGs (GSDMC, CASP4 and NLRP1) and the TCGA-PDAC cohort was classified into low and high-risk subgroups. It is noteworthy that the high-risk group showed significantly higher tumor proliferation rates and was proved to be highly correlated with oxaliplatin resistance. Further experiments suggested that overexpression of GSDMC promoted the proliferation and oxaliplatin resistance of PA-TU-8988T cells in vitro and vivo, while downregulation of GSDMC showed opposite effects in CFPAC-1 cells. Finally, we found that the activation of pentose phosphate pathway (PPP) was the mechanism by which GSDMC overexpression promoted the proliferation and oxaliplatin resistance of pancreatic cancer cells. CONCLUSIONS: In this study, we found that higher pyroptosis activity is associated with worse prognosis and oxaliplatin resistance of PDAC patients. In addition, as a core effector of pyroptosis, GSDMC promoted proliferation and oxaliplatin resistance of pancreatic cancer cells, which will provide new therapeutic target for PDAC patients.


Assuntos
Adenocarcinoma , Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Humanos , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/genética , Oxaliplatina/farmacologia , Oxaliplatina/uso terapêutico , Piroptose/genética , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/genética , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Carcinoma Ductal Pancreático/tratamento farmacológico , Carcinoma Ductal Pancreático/genética , Gasderminas , Biomarcadores Tumorais/metabolismo
6.
Ann Pharmacother ; : 10600280241283966, 2024 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-39342424

RESUMO

BACKGROUND: The optimal dosing of intravenous ganciclovir in patients receiving sustained low-efficiency dialysis (SLED) remains unclear. OBJECTIVE: The primary objective is to characterize the dosing of ganciclovir for treating and preventing cytomegalovirus (CMV) in Solid Organ Transplant Recipients receiving SLED. The secondary objective is to evaluate the safety and efficacy of the dosing practices described in this study. METHODS: Retrospective review of electronic medical records from solid organ transplant recipients (SOTRs) admitted to the Medical Surgical Intensive Care Unit at the Toronto General Hospital (TGH) between November 28, 2016, and September 1, 2021, was conducted. Patients concurrently receiving ganciclovir and SLED were included. RESULTS: Among the 27 encounters for CMV prevention, 18 patients underwent 8-hour SLED, 6 underwent 24-hour SLED, and 3 received other SLED durations. Most patients (80%) on 8-hour SLED began ganciclovir at 2.5 mg/kg/d, whereas 80% of those on 24-hour SLED started at 5 mg/kg/d. No breakthrough viremia occurred at 5 mg/kg/d, with 1 instance at 2.5 mg/kg/d. Cytopenia rates were higher at 5 mg/kg/d (33% vs 20%). For treatment (n = 20), 16 patients underwent 8-hour SLED, 2 underwent 24-hour SLED, and 2 underwent 12-hour SLED. Most (75%) on 8-hour SLED started at 2.5 mg/kg/d, whereas all on 24-hour SLED began at 5 mg/kg/d. Viral eradication rates were 75% and 60% at 2.5 and 5 mg/kg/d, respectively, with higher cytopenia rates at 5 mg/kg/d (37.5% vs 0%). Dose adjustments were primarily in response to refractory disease or cytopenia. CONCLUSION AND RELEVANCE: At our institution, ganciclovir dosing patterns suggest that for patients requiring 8-hour SLED, there is clinician comfort in using 2.5 mg/kg/d for prevention and 5 mg/kg/d for treatment. In 24-hour SLED, 5 mg/kg/d may be considered for prevention. Higher doses may be considered for CMV treatment; however, we found greater variability in the dosing practices for these patients. Further research with larger sample sizes and ganciclovir drug-level assessments is needed to optimize dosing strategies for CMV treatment.

7.
Artigo em Inglês | MEDLINE | ID: mdl-38642117

RESUMO

The neurobiological mechanism of borderline personality disorder (BPD) in adolescents remains unclear. The study aimed to assess the alterations in neural activity within prefrontal cortex in adolescents with BPD and investigate the relationship of prefrontal activity with emotional regulation and cognitive function. This study enrolled 50 adolescents aged 12-17 years with BPD and 21 gender and age-matched healthy control (HC) participants. Study assessment for each participant included a brain resting-state functional MRI (rs-fMRI), clinical assessment questionnaires such as Borderline Personality Features Scale (BPFS), Difficulties in Emotion Regulation Scale (DERS), Ottawa Self-Injury Inventory and Childhood Trauma Questionnaire (CTQ) and cognitive testing with Stroop Color-Word Test (SCWT). Fractional amplitude of low-frequency fluctuations (fALFF) and seed-based functional connectivity (FC) were obtained from rs-fMRI analysis. Correlation analysis was also performed to evaluate the associations of the neuroimaging metrics such as fALFF and FC with clinical assessment questionnaire and cognitive testing scores. Adolescents with BPD showed increased fALFF values in the right inferior frontal gyrus and decreased activity in the left middle frontal gyrus as compared to the HC group (p < 0.05, cluster size ≥ 100, FWE correction). In adolescents with BPD, increased fALFF in the right inferior frontal gyrus was related to the BPFS (emotional dysregulation), DERS-F (lacking of emotional regulation strategies) and Ottawa Self-Injury Inventory-4 C scores (internal emotional regulation function of self-injurious behavior). The reduced fALFF in the left middle frontal gyrus was associated with the SCWT-A (reading characters) and the SCWT-B (reading color) scores. Additionally, the fALFF values in the left middle frontal gyrus and the right inferior frontal gyrus were related to the CTQ-D (emotional neglect) (p < 0.05). The left middle frontal gyrus exhibited increased FC with the right hippocampus, left inferior temporal gyrus and right inferior frontal gyrus (voxel p < 0.001, cluster p < 0.05, FWE correction). The increased FC between the left middle frontal gyrus and the right hippocampus was related to the SCWT-C (cognitive flexibility) score. We observed diverging changes in intrinsic brain activity in prefrontal cortex, and neural compensatory changes to maintain function in adolescents with BPD. In addition, decreased neural function was closely associated with emotional dysregulation, while increased neural function as indicated by brain activity and FC was associated with cognitive dysfunction. These results indicated that alterations of intrinsic brain activity may be one of the underlying neurobiological markers for clinical symptoms in adolescents with BPD.

8.
Surg Innov ; 31(1): 16-25, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37899450

RESUMO

BACKGROUND: Breast-conserving surgery combined with oncoplastic breast surgery has become the standard surgical treatment for early breast cancer. OBJECTIVE: The purpose of this study was to investigate the safety and efficacy of the thoracodorsal artery perforator flap (TDAPF) in breast-conserving reconstruction of T2 breast cancer. METHODS: Thirty patients with T2 breast cancer admitted to our hospital from January 2019 to December 2020 were enrolled to receive pedicled TDAPF for repairing breast defects after breast-conserving surgery. Intraoperative conditions, postoperative complications, and shape satisfaction after breast reconstruction were recorded. RESULTS: The operation was successfully completed in all 30 patients, with an operation time of 177.77 ± 24.39 min, bleeding of 44.17 ± 7.67 mL, and length of hospital stay of 5.23 ± .97 d. There was no deformity or seroma at the donor site. Breast shape recovered well after operation. After operation, one patient had fat liquefaction in the recipient site, which healed well after wound treatment. The incidence of postoperative complications was 3.33%. Postoperative follow-up lasted 16-28 months, with a median of 22 months. The Breast-Q score for breast satisfaction was 61.83 ± 12.87 at 6 months after operation, compared to 62.07 ± 11.78 before operation (P > .05). CONCLUSIONS: TDAPF, featuring a high survival rate, moderate flap area, fewer postoperative complications, and high satisfaction with breast shape after operation. For east asian women with moderate breast size, TDAPF is a safe, effective choice for repairing defects in breast-conserving surgery for T2 breast cancer.


Assuntos
Neoplasias da Mama , Mamoplastia , Retalho Perfurante , Lesões dos Tecidos Moles , Humanos , Feminino , Neoplasias da Mama/cirurgia , Retalho Perfurante/irrigação sanguínea , Retalho Perfurante/cirurgia , Mamoplastia/efeitos adversos , Artérias/cirurgia , Complicações Pós-Operatórias/epidemiologia , Resultado do Tratamento , Transplante de Pele , Lesões dos Tecidos Moles/cirurgia
9.
Nat Chem Biol ; 17(1): 39-46, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32989297

RESUMO

Protein kinases control nearly every facet of cellular function. These key signaling nodes integrate diverse pathway inputs to regulate complex physiological processes, and aberrant kinase signaling is linked to numerous pathologies. While fluorescent protein-based biosensors have revolutionized the study of kinase signaling by allowing direct, spatiotemporally precise kinase activity measurements in living cells, powerful new molecular tools capable of robustly tracking kinase activity dynamics across diverse experimental contexts are needed to fully dissect the role of kinase signaling in physiology and disease. Here, we report the development of an ultrasensitive, second-generation excitation-ratiometric protein kinase A (PKA) activity reporter (ExRai-AKAR2), obtained via high-throughput linker library screening, that enables sensitive and rapid monitoring of live-cell PKA activity across multiple fluorescence detection modalities, including plate reading, cell sorting and one- or two-photon imaging. Notably, in vivo visual cortex imaging in awake mice reveals highly dynamic neuronal PKA activity rapidly recruited by forced locomotion.


Assuntos
Técnicas Biossensoriais , Proteínas Quinases Dependentes de AMP Cíclico/genética , Miócitos Cardíacos/enzimologia , Neurônios/enzimologia , Imagem Óptica/métodos , Alprostadil/farmacologia , Animais , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Di-Hidroxifenilalanina/farmacologia , Dinoprostona/farmacologia , Corantes Fluorescentes/química , Expressão Gênica , Biblioteca Gênica , Genes Reporter , Peptídeo 1 Semelhante ao Glucagon/farmacologia , Células HEK293 , Células HeLa , Ensaios de Triagem em Larga Escala , Hipocampo/citologia , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Humanos , Camundongos , Microscopia de Fluorescência por Excitação Multifotônica , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/ultraestrutura , Neurônios/efeitos dos fármacos , Neurônios/ultraestrutura , Cultura Primária de Células , Transdução de Sinais
10.
J Cell Physiol ; 236(7): 5278-5292, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33452687

RESUMO

Osteoarthritis (OA) is the most common joint disease. The surface of joint cartilage is a defensive and first affected structure of articular cartilage (AC) during the pathogenesis of OA. Alk5 signaling is critical for maintaining AC homeostasis, however, the role and underlying mechanism for the involvement of Alk5 signaling in the phenotypes of articular cartilage stem cells (ACSCs) at the surface of AC is still unclear. The role of Alk5 in OA development was explored using an ACSCs-specific Alk5-deficient (cKO) mouse model. Alterations in cartilage structure were evaluated histologically. Senescence was detected by SA-ß-gal, while reactive oxygen species (ROS), MitoTracker, and LysoTracker staining were used to detect changes related to senescence. In addition, mice were injected intra-articularly with ganciclovir to limit the detrimental roles of senescent cells (SnCs). Alk5 cKO mice showed a decreased number of the slow-cell cycle cells and less lubricant secretion at the surface accompanied with drastically accelerated cartilage degeneration under ageing and surgically induced OA conditions. Further studies showed that Alk5 deficient ACSCs exhibited senescence-like manifestations including decreased proliferation and differentiation, more SA-ß-gal-positive cells and ROS production, as well as significantly swollen mitochondria and lysosome breakdown. We further found that local limitation of the detrimental roles of SnCs can attenuate the development of posttraumatic OA. Taken together, our findings suggest that Alk5 signaling acts as an important regulator of the SnCs in the superficial layer during AC maintenance and OA initiation.


Assuntos
Cartilagem Articular/metabolismo , Senescência Celular/fisiologia , Osteoartrite/metabolismo , Receptor do Fator de Crescimento Transformador beta Tipo I/metabolismo , Células-Tronco/metabolismo , Animais , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Cartilagem Articular/patologia , Camundongos , Camundongos Knockout , Osteoartrite/patologia , Transdução de Sinais/fisiologia , Fator de Crescimento Transformador beta/metabolismo
11.
J Recept Signal Transduct Res ; 41(2): 145-152, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32757700

RESUMO

PURPOSE: This paper aims to assess the role of CEMIP in human breast cancer, and explore the potential mechanisms of CEMIP in the progression of breast cancer. METHODS: We examined the expression levels of CEMIP in breast cancer tissues and cell lines by quantitative PCR and immunohistochemical assays. The effects of CEMIP on the proliferation, migration and invasion of breast cancer cells were determined through colony formation assay, flow cytometry (FCM) assay, wound healing assay and transwell assay, respectively. Additionally, the downstream targets of CEMIP were examined by Immunoblot assays. RESULTS: CEMIP expression was upregulated in both breast cancer tissues and cell lines. Patients with higher CEMIP expression displayed shorter survival time than those with lower expression. In addition, CEMIP knockdown inhibited breast cancer cell proliferation, migration, and invasion in vitro. Our data further confirmed that CEMIP promoted the proliferation and migration of breast cancer cells via GRP78-STAT3 axis. CONCLUSION: Our data demonstrated the role of CEMIP in breast cancer was associated with STAT3 signaling pathway. Therefore, CEMIP might act as a possible therapeutic target for breast cancer.


Assuntos
Neoplasias da Mama/genética , Proteínas de Choque Térmico/genética , Hialuronoglucosaminidase/genética , Fator de Transcrição STAT3/genética , Apoptose/genética , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Chaperona BiP do Retículo Endoplasmático , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Transdução de Sinais
12.
Int J Hyperthermia ; 38(1): 1242-1250, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34402370

RESUMO

OBJECTIVE: This study aimed to evaluate the efficacy and safety of ultrasound-guided microwave ablation combined with glucocorticoid therapy for treating idiopathic granulomatous mastitis (IGM). METHODS: From June 2017 to March 2020, 50 consecutive patients diagnosed with IGM using puncture histology were included. All patients received prednisone and ultrasound-guided microwave ablation and were closely monitored for 12-15 months. RESULTS: A total of 222 lesions in 50 patients were ablated. The results indicated that 78% of cases were cured within 12 months and an additional 20% were cured within 15 months; the recurrence rate was 2%. The clinical and pathological remission rate of the entire group was 98%. The main postoperative complications were local pain, skin ulcerations and sinus formation, skin and areola heat damage, subcutaneous congestion, and fat liquefaction, all of which were conservatively treated. CONCLUSION: Microwave ablation combined with glucocorticoid therapy was safe and effective for the treatment of IGM, with a low recurrence rate. In addition, the cosmetic appearance of the affected breast was preserved with little trauma. Therefore, microwave ablation is a viable method that can be successfully applied in clinical practice.


Assuntos
Mastite Granulomatosa , Ablação por Radiofrequência , Feminino , Mastite Granulomatosa/diagnóstico por imagem , Mastite Granulomatosa/tratamento farmacológico , Mastite Granulomatosa/cirurgia , Humanos , Micro-Ondas , Resultado do Tratamento , Ultrassonografia de Intervenção
13.
J Obstet Gynaecol Res ; 47(9): 3362-3369, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34110061

RESUMO

AIM: To provide medical evidence for the feasibility and clinical efficacy of microwave ablation (MWA) in the treatment of benign breast lesions, especially multiple benign breast lesions. METHODS: We included patients with multiple benign breast lesions who were seen at our hospital during the period from June 2016 to December 2017. After signed informed consent had been obtained and core breast biopsy, all included patients who underwent MWA at 2450 MHz with 30 W power adjustment. Postoperative follow-up was performed for 1 year, at 3-month intervals. Ablation time, complications, volume-reduction ratio (VRR), and lesion residuals after ablation were analyzed. RESULTS: We ultimately included 1274 lesions from 164 cases in the study with a median follow-up time of 13.6 months. For a single lesion, the median ablation time was 36 s. Pain, fat liquefaction, and skin scalding were the most common complications. The overall complete disappearance rate was 92.1% (1173/1274), with the highest complete disappearance rate observed among those patients with lesions <10 mm in diameter (942/968, 97.3%). For lesions that persisted at 12 months after MWA, the average the volume-reduction ratio (VRR) was 87.5%. CONCLUSIONS: For multiple benign lesions of the breast, especially lesions with the longest diameter <10 mm, ultrasound guided MWA is a minimally invasive, relatively quick therapeutic strategy associated with accuracy in treatment and few complications.


Assuntos
Ablação por Cateter , Ablação por Radiofrequência , Humanos , Micro-Ondas , Estudos Prospectivos , Resultado do Tratamento , Ultrassonografia de Intervenção
14.
J Recept Signal Transduct Res ; 40(5): 410-418, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32314642

RESUMO

Objectives: Wolf-Hirschhorn syndrome candidate gene-1 (WHSC1) is highly expressed in various malignant tumors. We investigated the correlation and regulatory pathway of WHSC1 in the progression of breast cancer (BC).Methods: The expression and distribution of WHSC1 in the BC tissues and cell lines were determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemical staining. Spearman correlation analysis demonstrated the correlation between WHSC1 high expression level and the clinical characteristics of BC patients. The effects of WHSC1 on the proliferation, apoptosis, migration and invasion of BC cells were analyzed by cell transfection, MTT, colony formation, scratch assay, and transwell. Furthermore, the expression of Forkhead box M1 (FoxM1) and the location of ß-catenin were detected by qRT-PCR and western blot.Results: Firstly, WHSC1 expression was up-regulated in BC tissues and cell lines. The high expression of WHSC1 in BC is associated with the tumor size (p = 0.027), metastasis (p = 0.018) and pathological stages (p = 0.025) of the BC patients. The knockdown of WHSC1 inhibited the growth, proliferation migration, invasion and EMT of BC cell lines. Furthermore, WHSC1 could promote the expression of FoxM1 in BC cells and tissues. WHSC1 enhanced the expression of FoxM1, and promoted the nuclear localization of ß-catenin, and thus activated the downstream genes expression of Wnt/ß-catenin signaling pathway to regulate the development of BC.Conclusion: In summary, our study elucidates the correlation and specific regulatory mechanism between WHSC1 and the progression of BC, thus implying that WHSC1 may function as molecular diagnosis, prognosis and molecular targeted therapy of BC.


Assuntos
Neoplasias da Mama/genética , Proliferação de Células/genética , Proteína Forkhead Box M1/genética , Histona-Lisina N-Metiltransferase/genética , Proteínas Repressoras/genética , Neoplasias da Mama/patologia , Transição Epitelial-Mesenquimal/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Via de Sinalização Wnt/genética
15.
Zhongguo Dang Dai Er Ke Za Zhi ; 21(1): 6-10, 2019 Jan.
Artigo em Zh | MEDLINE | ID: mdl-30675856

RESUMO

OBJECTIVE: To investigate the incidence of neonatal asphyxia and possible contributing factors for the development of severe asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture, China. METHODS: A total of 16 hospitals in Hubei Enshi Tujia and Miao Autonomous Prefecture were selected as research centers. A retrospective analysis was performed for the clinical data of 22 294 live births in these 16 hospitals from January to December, 2016 to investigate the incidence rate of neonatal asphyxia and possible contributing factors for the development of severe asphyxia. RESULTS: Of the 22 294 neonates born alive, 733 (3.29%) were diagnosed with neonatal asphyxia, among whom 627 had mild asphyxia and 106 had severe asphyxia. The neonates with low maternal education level, maternal anemia during pregnancy, chorioamnionitis, abnormal amniotic fluid, abnormal umbilical cord, placenta previa, placental abruption, Tujia Minority, preterm birth, and low birth weight had a higher incidence of severe asphyxia (P<0.05). CONCLUSIONS: The incidence rate of neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture is higher. Low maternal education level, maternal anemia during pregnancy, chorioamnionitis, abnormal amniotic fluid, abnormal umbilical cord, placenta previa, placental abruption, Tujia Minority, preterm birth, and low birth weight may be related to the development of severe neonatal asphyxia.


Assuntos
Asfixia Neonatal , Asfixia Neonatal/epidemiologia , China , Humanos , Incidência , Recém-Nascido , Estudos Retrospectivos
16.
Cogn Process ; 19(1): 133-139, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28986700

RESUMO

Previous studies have shown that music is a powerful means to convey affective states, but it remains unclear whether and how social context shape the intensity and quality of emotions perceived in music. Using a within-subject design, we studied this question in two experimental settings, i.e. when subjects were alone versus in company of others without direct social interaction or feedback. Non-vocal musical excerpts of the emotional qualities happiness or sadness were rated on arousal and valence dimensions. We found evidence for an amplification of perceived emotion in the solitary listening condition, i.e. happy music was rated as happier and more arousing when nobody else was around and, in an analogous manner, sad music was perceived as sadder. This difference might be explained by a shift of attention in the presence of others. The observed interaction of perceived emotion and social context did not differ for stimuli of different cultural origin.


Assuntos
Emoções/fisiologia , Felicidade , Música/psicologia , Adolescente , Adulto , Nível de Alerta/fisiologia , Atenção/fisiologia , Percepção Auditiva/fisiologia , Feminino , Humanos , Masculino , Adulto Jovem
17.
bioRxiv ; 2024 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-38586003

RESUMO

The protein kinase C (PKC) family of serine/threonine kinases, which consist of three distinctly regulated subfamilies, have long been established as critical for a variety of cellular functions. However, how PKC enzymes are regulated at different subcellular locations, particularly at emerging signaling hubs such as the ER, lysosome, and Par signaling complexes, is unclear. Here, we present a sensitive Excitation Ratiometric (ExRai) C Kinase Activity Reporter (ExRai-CKAR2) that enables the detection of minute changes in subcellular PKC activity. Using ExRai-CKAR2 in conjunction with an enhanced diacylglycerol (DAG) biosensor capable of detecting intracellular DAG dynamics, we uncover the differential regulation of PKC isoforms at distinct subcellular locations. We find that G-protein coupled receptor (GPCR) stimulation triggers sustained PKC activity at the ER and lysosomes, primarily mediated by Ca2+ sensitive conventional PKC (cPKC) and novel PKC (nPKC), respectively, with nPKC showing high basal activity due to elevated basal DAG levels on lysosome membranes. The high sensitivity of ExRai-CKAR2, targeted to either the cytosol or Par-complexes, further enabled us to detect previously inaccessible endogenous atypical PKC (aPKC) activity in 3D organoids. Taken together, ExRai-CKAR2 is a powerful tool for interrogating PKC regulation in response to physiological stimuli.

18.
J Colloid Interface Sci ; 671: 1-14, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-38788420

RESUMO

The widespread contamination of hexavalent chromium (Cr(VI)), pharmaceuticals and personal care products (PPCPs), and dyes is a growing concern. necessitating the development of convenient and effective technologies for their removal. Copper(I) phenylacetylide (PhC2Cu) has emerged as a promising photocatalyst for environmental remediation. In this study, we introduced a functional Cu-O bond into PhC2Cu (referred to as OrPhC2Cu) by creatively converting the adsorbed oxygen on the surface of PhC2Cu into a Cu-O bond to enhance the efficiency of Cr(VI) photoreduction, PPCPs photodegradation, and dyes photodegradation through a facile vacuum activating method. The incorporation of the Cu-O bond optimized the electron structure of OrPhC2Cu, facilitating exciton dissociation and charge transfer. The exciton dissociation behavior and charge transfer mechanism were systematically investigated for the first time in the OrPhC2Cu system by photoelectrochemical tests, fluorescence and phosphorescence (PH) techniques, and density functional theory (DFT) calculations. Remarkably, the enhanced visible-light response of OrPhC2Cu improved photon utilization and significantly promoted the generation of reactive species (RSs), leading to the highly efficient Cr(VI) photoreduction (98.52% within 25 min) and sulfamethazine photodegradation (94.65% within 60 min), with 3.91 and 5.23 times higher activity compared to PhC2Cu. Additionally, the photocatalytic efficiency of OrPhC2Cu in degrading anionic dyes surpassed that of cationic dyes. The performance of the OrPhC2Cu system in treating electroplating effluent or natural water bodies suggests its potential for practical applications.

19.
Asian J Surg ; 46(1): 254-259, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35393222

RESUMO

BACKGROUND: The breasts of Oriental women are characterized by an obvious scar constitution and a relatively small mammary gland volume. Thus, plastic surgery, which is now popular in the West, is not suitable for most patients in China, and Chinese surgeons are searching for symmetrical plastic surgery options that are suitable for patients with breast tumors, unilateral breast implants and an obvious scar constitution. METHODS: Between January 2016 and December 2019, 15 patients underwent contralateral breast overlapped reconstruction (COBOR) at the Affiliated Hospital of Putian University. We assessed their clinicopathological data, complications, cosmetic satisfaction and quality of life. RESULTS: The mean age was 41.6 years (range, 31-54 years), the average BMI was 24.36 kg/m2 (range, 20.3-28.4 kg/m2), the most common tumor location was the upper outer quadrant (n = 9), the mean preoperative tumor size was 21.11 mm (range, 7-42 mm), and 4 patients underwent neoadjuvant chemotherapy. The cancer grades and histological types were as follows: G3 nonspecial type (NST), 3 cases; G2 NST, 6 cases; G2 lobular carcinoma, 1 case; and ductal carcinoma in situ (DCIS), 5 cases. The nipple margin was negative in all of these cases. Among them, there was 1 case of poor wound healing caused by subcutaneous fat liquefaction around the incision. In another case, partial nipple necrosis occurred on the affected side due to an insufficient nipple blood supply after the operation and healed after debridement and dressing changes. There were no cases of tumor recurrence during the mean follow-up of 22.53 months (range, 11-47 months). The BREAST-Q scores showed that COBOR provided good patient satisfaction. CONCLUSION: For Oriental patients with small breasts, COBOR, which results in fewer scars, good symmetry and good satisfaction, is an effective and safe surgical method. However, larger studies with longer follow-up periods are needed to obtain more reliable postoperative results.


Assuntos
Neoplasias da Mama , Mamoplastia , Neoplasias Unilaterais da Mama , Feminino , Humanos , Adulto , Mastectomia/métodos , Neoplasias da Mama/cirurgia , Neoplasias da Mama/patologia , Neoplasias Unilaterais da Mama/patologia , Neoplasias Unilaterais da Mama/cirurgia , Seguimentos , Cicatriz , Qualidade de Vida , Mamoplastia/métodos , Mamilos/cirurgia , Estudos Retrospectivos
20.
J Hazard Mater ; 458: 132009, 2023 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-37429189

RESUMO

The widespread use of sulfonamides (SAs) in animals and human infections has raised significant concerns regarding their presence in ambient waterways and potential for inducing antimicrobial resistance. Herein, we report on the capacity of ferrate (VI) (FeVIO42-, Fe(VI)) to facilitate the photocatalytic degradation of sulfamethazine (SMT) via bismuth tungstate (Bi2WO6, BWO) under blue LED light (Vis/BWO/Fe(VI)) exposure, at rates that were 45-fold faster than BWO photocatalysis. Both the stepwise and time-series addition of Fe(VI) contributed to the degradation. Multiple lines of evidence confirmed that the common reactive species (RSs) in BWO-based photocatalytic systems and Fe(VI)-involved systems (e.g., •OH/h+, O2•-, 1O2 and Fe(V)/Fe(IV)) played subtle roles in our study system. Herein, for the first time, it was discovered that the precursor complex (BWO-Fe(V)/Fe(IV)* )) was the main contributor to induce electron transfer of SAs through the "conductive bridge" effect of BWO. The studied system was able to effectively degrade SMT in synthetic hydrolyzed urine (SHU) with low interference from background substances in water. This work not only offers a novel facilitation strategy for BWO, but also holds a great application prospect for contamination remediation in urine.


Assuntos
Elétrons , Sulfonamidas , Humanos , Oxirredução , Transporte de Elétrons , Luz
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