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1.
BMC Nurs ; 23(1): 426, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38918789

RESUMO

BACKGROUND: The experiences and perceptions of geriatric specialist nurses are pivotal to understanding the complexities of managing delirium and to developing effective nursing interventions. This qualitative study aims to explore these experiences and perceptions to inform the enhancement of clinical geriatric nursing and care practices. METHODS: Utilizing a qualitative exploratory design, this research engaged a convenience sample of geriatric specialist nurses at a tertiary hospital in Shanghai, China through focus groups and semi-structured interviews. Data were rigorously analyzed using Colaizzi's phenomenological method, which facilitated the identification of themes that emerged from the narratives of the geriatric specialist nurses. RESULTS: The thematic analysis yielded three major themes that encapsulate the nurses' experiences and perceptions. Theme 1: Understanding of Delirium, highlighted the nurses' awareness of the condition's significance, yet it was often deprioritized due to the pressing demands of managing more acute and immediately life-threatening conditions. Theme 2: Barriers in Application, brought to light the multifaceted challenges faced by nurses, including language barriers, the frequency and consistency of delirium assessments, the social determinants of health, and the nurses' own competencies in assessment. Theme 3: Evolution of Nursing Approaches, detailed the adaptive strategies employed by nurses, such as managing nursing adverse events, improving communication with patients' families, and adopting a proactive stance towards long-term patient outcomes. CONCLUSIONS: The findings suggest that while geriatric specialist nurses recognize the importance of delirium assessment, there are several barriers to effective application. The study underscores the imperative for the advancement of more refined delirium assessment and care protocols, tailored to address the unique requirements of geriatric nursing care.

2.
BMC Genomics ; 24(1): 126, 2023 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-36932328

RESUMO

BACKGROUND: Late embryogenesis abundant (LEA) proteins play an important role in dehydration process of seed maturation. The seeds of Panax notoginseng (Burkill) F. H. Chen are typically characterized with the recalcitrance and are highly sensitive to dehydration. However, it is not very well known about the role of LEA proteins in response to dehydration stress in P. notoginseng seeds. We will perform a genome-wide analysis of the LEA gene family and their transcriptional responses to dehydration stress in recalcitrant P. notoginseng seeds. RESULTS: In this study, 61 LEA genes were identified from the P. notoginseng genome, and they were renamed as PnoLEA. The PnoLEA genes were classified into seven subfamilies based on the phylogenetic relationships, gene structure and conserved domains. The PnoLEA genes family showed relatively few introns and was highly conserved. Unexpectedly, the LEA_6 subfamily was not found, and the LEA_2 subfamily contained 46 (75.4%) members. Within 19 pairs of fragment duplication events, among them 17 pairs were LEA_2 subfamily. In addition, the expression of the PnoLEA genes was obviously induced under dehydration stress, but the germination rate of P. notoginseng seeds decreased as the dehydration time prolonged. CONCLUSIONS: We found that the lack of the LEA_6 subfamily, the expansion of the LEA_2 subfamily and low transcriptional levels of most PnoLEA genes might be implicated in the recalcitrant formation of P. notoginseng seeds. LEA proteins are essential in the response to dehydration stress in recalcitrant seeds, but the protective effect of LEA protein is not efficient. These results could improve our understanding of the function of LEA proteins in the response of dehydration stress and their contributions to the formation of seed recalcitrance.


Assuntos
Panax notoginseng , Panax notoginseng/genética , Panax notoginseng/metabolismo , Desidratação/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Filogenia , Sementes/metabolismo , Desenvolvimento Embrionário , Regulação da Expressão Gênica de Plantas
3.
Anal Chem ; 95(5): 2628-2632, 2023 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-36705511

RESUMO

A novel microfiber-like biohydrogel was fabricated by a facile approach relying on electroactive bacteria-induced graphene oxide reduction and confined self-assembly in a capillary tube. The microfiber-like biohydrogel (d = ∼1 mm) embedded high-density living cells and activated efficient electron exchange between cells and the conductive graphene network. Further, a miniature whole-cell electrochemical biosensing system was developed and applied for fumarate detection under -0.6 V (vs Ag/AgCl) applied potential. Taking advantage of its small size, high local cell density, and excellent electron exchange, this microfiber-like biohydrogel-based sensing system reached a linear calibration curve (R2 = 0.999) ranging from 1 nM to 10 mM. The limit of detection obtained was 0.60 nM, which was over 1300 times lower than a traditional biosensor for fumarate detection in 0.2 µL microdroplets. This work opened a new dimension for miniature whole-cell electrochemical sensing system design, which provided the possibility for bioelectrochemical detection in small volumes or three-dimensional local detection at high spatial resolutions.


Assuntos
Técnicas Biossensoriais , Grafite , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Bactérias , Fumaratos , Condutividade Elétrica , Limite de Detecção
4.
Sens Actuators B Chem ; 379: 133252, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36590306

RESUMO

SARS-CoV-2, a highly transmissible and mutagenic virus, made huge threats to global public health. The detection strategies, which are free from testing site requirements, and the reagents and instruments are portable, are vital for early screening and play a significant role in curbing the spread. This work proposed a silver-coated glass slide (SCGS)/DNA walker based on a dual targets-triggering mechanism, enzyme-catalyzed amplification, and smartphone data analysis, which build a portable visual detection strategy for the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) gene. By this method, the detection was reflected by the ultraviolet absorbance changes and visible color changes to the naked eye which was analyzed by Red-Green-Blue (RGB) data analysis via smartphone within 30 min, simplifying the detection process and shortening the detection time. Meanwhile, the dual targets-triggering mechanism and dual signal amplification strategy ensured detection specificity and sensitivity. Further, the practicability was verified by the detection of the real sample which provided this method an application potential in SARS-CoV-2 rapid detection.

5.
Int J Mol Sci ; 24(4)2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36835274

RESUMO

Ion exchange resins are suitable as carriers for immobilized enzymes because of their stable physicochemical properties, appropriate particle size and pore structure, and lower loss in continuous operation. In this paper, we report the application of the Ni-chelated ion exchange resin in the immobilization of His-tagged enzyme and protein purification. Acrylic weak acid cation exchange resin (D113H) was selected from four cationic macroporous resins that could chelate the transition metal ion Ni. The maximum adsorption capacity of Ni was ~198 mg/g. Phosphomannose isomerase (PMI) can be successfully immobilized on Ni-chelated D113H from crude enzyme solution through chelation of transition metal ions with the His-tag on the enzyme. The maximum amount of immobilized PMI on the resin was ~143 mg/g. Notably, the immobilized enzyme showed excellent reusability and maintained 92% of its initial activity with 10 cycles of catalytic reaction. In addition, PMI was successfully purified using an affinity chromatography column prepared by Ni-chelated D113H, which showed the potential for the immobilization and purification process to be realized in one step.


Assuntos
Enzimas Imobilizadas , Resinas de Troca Iônica , Proteínas , Quelantes/química , Cromatografia de Afinidade/métodos , Enzimas Imobilizadas/química , Indicadores e Reagentes , Proteínas/isolamento & purificação
6.
Chembiochem ; 23(4): e202100497, 2022 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-34958513

RESUMO

Self-stable precipitation polymerization was used to prepare an enzyme-immobilized microsphere composite. Phosphomannose isomerase (PMI) with His-tag was successfully immobilized on Ni2+ charged pyridine-derived particles. The maximum amount of PMI immobilized on such particles was ∼184 mg/g. Compared with free enzyme, the activity of the immobilized enzymes was significantly improved. In addition, the immobilized enzymes showed a much better thermostability than free enzymes. At the same time, the immobilized enzymes can be reused for multiple reaction cycles. We observed that the enzyme activity did not decrease significantly after six cycles. We conclude that the pyridine-derived particles can be used to selectively immobilize His-tagged enzymes, which can couple the enzyme purification and catalysis steps and improve the efficiency of enzyme-catalyzed industrial processes.


Assuntos
Quelantes/metabolismo , Histidina/metabolismo , Manose-6-Fosfato Isomerase/metabolismo , Nanopartículas/metabolismo , Níquel/metabolismo , Linhagem Celular Tumoral , Quelantes/química , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Histidina/química , Humanos , Manose-6-Fosfato Isomerase/química , Nanopartículas/química , Níquel/química
7.
BMC Psychiatry ; 21(1): 363, 2021 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-34284741

RESUMO

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic profoundly impacts on mental health, yet it is still unclear whether COVID-19 distress makes people more vulnerable to suicidal behavior. The present study aims to examine the association between COVID-19 related psychological distress and risk for suicide attempt, and moderators of this association, among hotline callers. METHODS: This case-control study was conducted at the largest psychological support hotline in China. Hotline callers who sought help for psychological distress and reported whether or not they attempted suicide in the last 2 weeks (recent suicide attempt) were analyzed. The primary predictor of recent suicide attempt was the presence or absence of COVID-19 related psychological distress. Demographic variables and common risk and protective factors for suicidal behavior were also studied. Callers with COVID-19 related distress (COVID-19 callers) and those without such distress (non-COVID-19 callers) were compared on these variables. Recent suicide attempt was regressed on COVID-19 related distress and the other variables, and significant interaction terms of aforementioned predictors by COVID-19 related distress, to identify variables that moderate the association of COVID-19 related distress and recent suicide attempt. RESULTS: Among 7337 included callers, there were 1252 COVID-19 callers (17.1%) and 6085 non-COVID-19 callers (82.9%). The COVID-19 callers were less likely to report recent suicide attempt (n = 73, 5.8%) than the non-COVID-19 callers (n = 498, 8.2%, P = 0.005). The COVID-19 callers were also less likely to have high scores on depressive symptoms (22.6% vs 26.3%, P < 0.001) and psychological distress (19.5% vs 27.3%, P < 0.001), and were more likely to have high hopefulness scores (46.5% vs 38.0%, P < 0.001). Tests of moderating effects showed that acute life events were associated with one-half lower risk (P = 0.021), and a trend that suicide attempt history was associated with two-thirds greater risk (P = 0.063) for recent suicide attempt, among COVID-19 callers than non-COVID-19 callers. CONCLUSIONS: The COVID-19 calls are from individuals with lower suicide-related risk compared to more typical callers. Acute stressful life events provided a key context for suicide attempt in non-COVID-19 callers, i.e., more typical calls.


Assuntos
COVID-19 , Angústia Psicológica , Estudos de Casos e Controles , China/epidemiologia , Linhas Diretas , Humanos , SARS-CoV-2 , Tentativa de Suicídio
8.
Biochem Biophys Res Commun ; 528(1): 179-185, 2020 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-32499111

RESUMO

Sirt1 involved in cellular aging and aging-related diseases, including osteoarthritis (OA). Our previous study showed Sirt1 played a role in the pathogenesis of OA, however, the underlying mechanisms are still poorly elicited. In this study, we investigated the role of Sirt1 in epigenetically regulating P53/P21 pathway in a Sirt1 loss model. Sirt1 deletion male mice (n = 10) with destabilization of the medial meniscus (DMM) were used to observe its role on OA development. Then, the relationships between SIRT1 and P53 were detected by Coimmunoprecipitation (CoIP), and the gain-off function of P53 gene was indicated by P53 activators and inhibitors in vitro. Finally, human cartilage samples from patients with OA were collected. Sirt1 deletion mice displayed a spontaneous OA development, manifesting severe chondrocytes hypertrophy markers MMP13 and ADAMTS5, highly expressed P53 and P21. Strikingly, surgery-induced meniscus injury promoted the OA pathogenesis and apoptosis in Sirt1 deficient mice. Ultimately, our CoIP data demonstrated that Sirt1 directly interacted with P53 in vitro. However inhibition of P53 alleviated OA progression. We also observed that chondrocyte apoptosis and P53 increased in osteoarthritis (OA) progression with a declining expression of Sirt1 in human cartilage. Loss of Sirt1 in cartilage led to accelerated OA pathogenesis via aberrant activation of p53/p21 mediated senescence associated secretory phenotype, hypertrophy and apoptosis.


Assuntos
Apoptose , Condrócitos/metabolismo , Condrócitos/patologia , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Epigênese Genética , Osteoartrite/genética , Sirtuína 1/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Idoso , Envelhecimento/patologia , Animais , Apoptose/genética , Cartilagem/patologia , Deleção de Genes , Humanos , Hipertrofia , Masculino , Camundongos Knockout , Pessoa de Meia-Idade , Osteoartrite/etiologia , Osteoartrite/patologia , Tíbia/patologia
9.
Surg Endosc ; 34(5): 2028-2034, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31312964

RESUMO

OBJECTIVE: The objective of the study is to evaluate the safety and feasibility of laparoscopic pancreaticoduodenectomy (LPD) in elderly patients by short-term surgical effects. METHODS: We retrospectively collected clinical data of 55 non-elderly patients (< 70 years), 27 elderly patients (≥ 70 years) underwent LPD, and 19 elderly patients underwent open pancreaticoduodenectomy (OPD) in biliopancreatic surgery department of Huadong Hospital, affiliated to Fudan University, from Jan 2015 to Jan 2018. Patients were divided into 3 groups: LPD aged < 70 years, LPD aged ≥ 70 years, and OPD aged ≥ 70 years, according to their age at admission and surgical approach in order to compare baseline characteristics and short-term surgical outcomes. RESULTS: Totally 101 patients were included in this study; 59 cases were male; 42 cases were female; mean age was 66 years old. Elderly LPD patients seemed to have higher overall morbidity (41% vs. 20%, P = 0.05) compared to non-elderly patients. This difference is even more significant in our multivariable analysis model with an odds ratio of 4.48 (95% CI 1.31-17.87, P = 0.018). The 90-days mortality, operative time, estimated blood loss (EBL), and post-operative hospital stay (POHS) were similar in two groups. Elderly LPD patients had less EBL and shorter POHS than elderly OPD patients. However, the mortality and morbidity rate were comparable in these two groups. CONCLUSIONS: Aging patients have higher overall morbidity than younger patients in LPD. However, for aging population who need to undergo pancreaticoduodenectomy, LPD might have some advantages over OPD.


Assuntos
Laparoscopia/métodos , Pancreaticoduodenectomia/métodos , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Estudos Retrospectivos
10.
Appl Microbiol Biotechnol ; 104(1): 241-255, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31735984

RESUMO

The co-culturing of Pleurotus eryngii var. ferulae and Rhodotorula mucilaginosa was confirmed in our previous studies to be an efficient strategy to improve laccase production by submerged fermentation. To determine the possible regulation principles underlying this behaviour, comparative transcriptomic analysis was performed on P. eryngii var. ferulae to investigate the differential expression of genes in co-culture. RNA-seq analysis showed that genes concerning xenobiotic biodegradation and expenditure of energy were upregulated. However, genes related to oxidative stress were downregulated. In addition, the transcription levels of laccase isoenzymes were not consistent in the co-culture system: 3 laccase genes (lacc1, lacc2, lacc12) were upregulated, and 3 laccase genes (lacc4, lacc6, lacc9) were downregulated. The enhancement in laccase activity can be due to upregulation of a laccase heterodimer encoded by the genes lacc2 and ssPOXA3a (or ssPOXA3b), whose expression levels were increased by 459% and 769% (or 585% for ssPOXA3b) compared with those of a control, respectively. ß-Carotene produced by R. mucilaginosa upregulated the transcription of lacc2 only. Combining these results with an analysis of cis-acting responsive elements indicated that four transcription factors (TFs) had potential regulatory effects on the transcription of laccase genes. It was supposed that TFa regulated lacc transcription by binding with methyl jasmonate and heat shock response elements. The expression of TFb, TFc, and TFd was regulated by ß-carotene. However, ß-carotene had no effect on TFa expression. These results provide a possible mechanism for the regulation of laccase gene transcription in the co-culture system and are also beneficial for the future intensification of fungal laccase production.


Assuntos
Regulação Fúngica da Expressão Gênica , Lacase/biossíntese , Pleurotus/genética , Rhodotorula/genética , Transcriptoma , Técnicas de Cocultura , Biologia Computacional , Regulação para Baixo , Fermentação , Proteínas Fúngicas/genética , Microbiologia Industrial , Lacase/genética , Pleurotus/metabolismo , Rhodotorula/metabolismo , Análise de Sequência de RNA , Regulação para Cima
11.
Aust N Z J Psychiatry ; 54(12): 1182-1191, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33050732

RESUMO

OBJECTIVES: This study aimed to assess the association of the quality scores of hotline psychological intervention and the reduction of subsequent suicidal acts among high suicidal risk callers. METHODS: High-risk callers at a national crisis hotline service in China were recruited and prospectively followed for up to 3 months after receiving a hotline psychological intervention. The quality of the intervention was evaluated by supervisors who listened to the tape-recorded calls using the Counseling Skills Rating Scale for Psychological Support Hotlines, which assessed three counseling domains: process, attitude and communication skill. The primary outcome was the occurrence of suicidal acts during the follow-up period. Secondary outcomes were before versus after changes during the intake intervention call in hopefulness, psychological stress and suicide intention reported by the callers. RESULTS: Over the 3-month follow-up, 45 of 778 high-risk callers reported 61 suicide attempts, and 3 other callers died by suicide. Subsequent suicidal act was significantly more common in callers classified as being at higher risk during the intake call. Higher scores on the quality of suicidality assessing of the Counseling Skills Rating Scale for Psychological Support Hotlines were associated with reduced risk of suicidal acts during follow-up (hazard ratio = 0.38, 95% confidence interval = [0.18, 0.85]). Higher scores on the communication skill domain were associated with increases in hopefulness (ß = 0.09) after the intervention, and higher scores on the counseling process domain (ß = -0.12) and higher suicidal risk scores (ß = -0.12) were associated with decreased suicide intention after intervention. CONCLUSION: Several characteristics of a hotline intervention for suicide prevention were associated with decreased risk of suicidal acts during follow-up. Intervention skill training for hotline operators should emphasize these specific counseling skills.


Assuntos
Linhas Diretas , Ideação Suicida , Intervenção em Crise , Humanos , Estudos Prospectivos , Intervenção Psicossocial
12.
J Clin Nurs ; 29(21-22): 4161-4170, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32757428

RESUMO

AIMS AND OBJECTIVES: To evaluate the mental health status, stressors and self-adjustment of nurses in isolation wards at different periods in Wuhan, China. BACKGROUND: Mental health issues easily occurred among the frontline medical workers of a major epidemic. However, the stressors and psychological adjustments experienced by nurses have not been well described. This is crucial to improving clinical quality and nursing safety and ensuring nurses' physical and psychological health. METHODS: We performed a cross-sectional prospective study using the Self Reporting Questionnaire-20, stressor and self-adjustment questionnaire administered to frontline nurses in Wuhan at two time points: after they had worked in isolation wards for 7-10 days (T1 ) and 2 months (T2 ). This paper complies with the STROBE reporting guideline for cross-sectional studies. RESULTS: T1 has 92 respondents, and T2 has 86. The positive rates of mental health problems were 26.09% and 9.30%, respectively, showing significantly different in the two periods. The main factors influenced mental health were self-perceived stress and only child status. The most common stressors were as follows: a large infected population, high infectivity; concerned about family's health status; high mortality if not treated in time (T1 ); and long duration of the epidemic, separate from family for a long time (T2 ). In terms of self-adjustment, 97.83%(T1 )/88.04%(T2 ) of nurses thought it was necessary, but 9(T1 ) /5(T2 ) chose to avoid addressing it, and 8(T1 ) /5(T2 ) utilised a professional psychological counselling hotline. CONCLUSIONS: Mental health problems among frontline nurses fighting COVID-19 need special attention, so administrators should offer timely counselling and strengthen effective psychosocial support to improve their mental resilience. RELEVANCE TO CLINICAL PRACTICE: This study surveyed the mental problems and self-adjustment status among nurses working Wuhan during the outbreak of COVID-19, to provide administrators with a scientific basis to effectively intervene.


Assuntos
COVID-19/enfermagem , Ajustamento Emocional , Recursos Humanos de Enfermagem Hospitalar/psicologia , Adulto , China , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Estudos Prospectivos , Fatores de Risco , SARS-CoV-2 , Inquéritos e Questionários
13.
J Environ Manage ; 270: 110904, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32721339

RESUMO

An efficient valorization of tea residues into value-added product was developed by Trametes versicolor in solid-state fermentation (SSF). The laccase production of 25.7 U/g dry substrate was obtained by optimizing culture medium and condition, resulting in a 4.0-fold increase compared to that of 6.4 U/g dry substrate under unoptimized condition. During the 7-day cultivation under SSF, 44.7%, 12.2% and 9.8% degradation occurred for lignin, hemicellulose and cellulose in tea residues, respectively. Laccase production reached 31.2 U/g dry substrate by the scaling-up culture in shallow tray system. The dry fermented tea residues were directly used as crude enzyme in the decolorization of malachite green. It possessed a decolorization rate of more than 95% within 120 min and remained 81.3% of decolorization capacity after 6 cycles. The present study provided a useful strategy for low-cost laccase production by SSF and it exhibited great potential for the application in dye decolorization.


Assuntos
Lacase , Trametes , Corantes , Fermentação , Lignina , Chá
14.
Fish Shellfish Immunol ; 80: 335-347, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29920382

RESUMO

Tumor necrosis factor receptor-associated factor 6 (TRAF6) acts as a central intracellular signal adapter molecule that mediates the tumor necrosis factor receptor superfamily and the interleukin-1 receptor/Toll-like receptor family in vertebrates and invertebrates. In the present study, HcTRAF6, a molluscan homologue of TRAF6 from Hyriopsis cumingii, has been cloned and identified. The entire open reading frame of HcTRAF6 was found to comprise a 1965-bp region that encodes a predicted protein of 654 amino acids, which contains conserved characteristic domains including a RING domain, two TRAF-type zinc finger domains, a typical coiled coil and the MATH domain. Phylogenetic analysis revealed that HcTRAF6 was aggregated closely with CsTRAF6 from Cyclina sinensis in the invertebrate cluster of mollusks. Further, qRT-PCR analysis showed that HcTRAF6 mRNA was extensively distributed in mussel tissues with a high expression in gills. After immune stimulation with Aeromonas hydrophila and lipopolysaccharides, the transcription of HcTRAF6 was obviously induced in the gills and hemocytes. In addition, significant fluctuation in HcTRAF6 expression was observed in the pearl sac, gills and hemocytes after mantle implantation. These findings confirmed its role in the alloimmune response. Dual-luciferase reporter assay showed that over-expression of HcTRAF6 could enhance the activity of the NF-κB reporter in a dose-dependent manner. Further, the RNA interference showed that the up-regulation of antimicrobial peptides in anti-bacterial infection was strongly suppressed in HcTRAF6-silenced mussels and that depletion of HcTRAF inhibited the elimination of A. hydrophila. All these findings together prove that HcTRAF6 functions as an efficient regulator in innate immune mechanisms against invading pathogens and the alloimmune mechanism after mantle implantation in H. cumingii.


Assuntos
Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/imunologia , Unionidae/genética , Unionidae/imunologia , Aeromonas hydrophila , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Brânquias/imunologia , Células HEK293 , Hemócitos/imunologia , Humanos , Imunidade Inata , Lipopolissacarídeos , Filogenia , RNA Interferente Pequeno/genética
15.
Fish Shellfish Immunol ; 59: 241-249, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27794460

RESUMO

The allograft inflammatory factor-1 (AIF-1) is one of the key factors associated with inflammatory response and immune defense. In the present study, we report the identification and characterization of AIF-1 from triangle sail mussel Hyriopsis cumingii (HcAIF-1). The full-length cDNA of HcAIF-1 consisted of a 5'-terminal untranslated region (UTR) of 80 bp, a 3'-UTR of 420 bp with a poly (A) tail, and an open reading frame of 444 bp encoding a polypeptide of 147 amino acids with two conserved EF-hand Ca2+-binding motifs. HcAIF-1 mRNA and protein were expressed in all examined tissues and showed higher mRNA expression levels were observed in immune tissues, especially hemocytes and mantle, and the highest protein expression level was in mantle. The expression level of HcAIF-1 mRNA was significantly upregulated in hemocytes 12-48 h after lipopolysaccharide challenge. After mantle tissue implantation, the expression level of this gene in pearl sac decreased significantly at 3-48 h (P < 0.01), and then was significantly upregulated at 96 h (P < 0.05) and recovered to the control level at 21-28 d. There was significant increase HcAIF-1 transcript abundance in hemocytes 96 h (P < 0.05) after mantle tissue implantation. The phagocytosis rate was significantly enhanced in hemocytes 3-24 h (P < 0.01) after the injection of recombinant HcAIF-1 protein. These findings suggest that HcAIF-1 is important in the underlying mechanism of the innate immune responses and pearl sac formation of H. cumingii.


Assuntos
Proteínas de Ligação ao Cálcio/genética , Regulação da Expressão Gênica , Imunidade Inata/genética , Unionidae/genética , Unionidae/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Ligação ao Cálcio/química , Proteínas de Ligação ao Cálcio/metabolismo , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Hemócitos/imunologia , Fagocitose , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência
16.
Fish Shellfish Immunol ; 56: 127-135, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27403593

RESUMO

Hyriopsis cumingii is the most important freshwater pearl mussel cultured in China. The operation for implantation is one necessary technical step for pearl culture. However, implantation-induced trauma results in a series of immune responses and can enable the invasion of pathogenic microbes. Lectin proteins are found widely in nature and play important roles in innate immunity. Galectins are members of the lectin superfamily and are characterized by one or several carbohydrate recognition domains (CRDs) that produce multiple sugar binding sites on the protein. Here we cloned and characterized the H. cumingii galectin gene HcGal1, which encodes a 312 amino acid galectin protein. The HcGal1 transcript was detected in all tested H. cumingii tissues and showed higher expression specifically in immune tissues. The significant upregulation of HcGal1 expression was observed after challenging the mussel with lipopolysaccharide or Gram-negative and Gram-positive bacteria. After implantation, significant downregulation of the HcGal1 transcript was noted in the mantle, hemocytes, and pearl sac in the acute-stress stage (0-24 h) and the stage of wound healing and pearl-sac formation (24 h-7 d). In addition, significant upregulation of HcGal1 expression was observed in the liver in the stage of wound healing and pearl-sac formation. In the pearl-secretion stage (7-35 d), the HcGal1 transcript levels returned to normal in all tested tissues. We also show that recombinantly expressed and purified HcGal1 can agglutinate some Gram-negative and Gram-positive bacteria. In addition, in vivo experiments showed that the recombinant protein HcGal1 could promote phagocytosis by hemocytes. Our data suggest that HcGal1 plays a role in innate immune responses involved in pathogen recognition and wound healing.


Assuntos
Fenômenos Fisiológicos Bacterianos/imunologia , Galectinas/genética , Regulação da Expressão Gênica , Imunidade Inata , Unionidae/genética , Unionidae/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Galectinas/química , Galectinas/metabolismo , Lipopolissacarídeos/farmacologia , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Unionidae/classificação , Unionidae/metabolismo
17.
Biochem Biophys Res Commun ; 455(3-4): 229-33, 2014 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-25449277

RESUMO

The function of dNTP hydrolase SAMHD1 as a viral restriction factor to inhibit the replication of several viruses in human immune cells was well established. However, its regulation and function in lung cancer have been elusive. Here, we report that SAMHD1 is down regulated both on protein and mRNA levels in lung adenocarcinoma compared to adjacent normal tissue. We also found that SAMHD1 promoter is highly methylated in lung adenocarcinoma, which may inhibit its gene expression. Furthermore, over expression of the SAMHD1 reduces dNTP level and inhibits the proliferation of lung tumor cells. These results reveal the regulation and function of SAMHD1 in lung cancer, which is important for the proliferation of lung tumor cells.


Assuntos
Adenocarcinoma/metabolismo , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/metabolismo , Proteínas Monoméricas de Ligação ao GTP/metabolismo , Adenocarcinoma/genética , Linhagem Celular Tumoral , Metilação de DNA , Regulação para Baixo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Neoplasias Pulmonares/genética , Proteínas Monoméricas de Ligação ao GTP/genética , Regiões Promotoras Genéticas , RNA Mensageiro/metabolismo , Proteína 1 com Domínio SAM e Domínio HD
18.
Anal Chim Acta ; 1299: 342406, 2024 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-38499412

RESUMO

The hybridization chain reaction (HCR), as one of the nucleic acid amplification technologies, is combined with fluorescence signal output with excellent sensitivity, simplicity, and stability. However, current HCR-based fluorescence sensing methods still have some defects such as the blocking effect of the HCR combination with fluorophores and the aggregation-caused quenching (ACQ) phenomenon of traditional fluorophores. Herein, a triplex DNA-based aggregation-induced emission probe (AIE-P) was designed as the fluorescent signal transduction, which is able to provide a new platform for HCR-based sensing assay. The AIE-P was synthesized by attaching the AIE fluorophores to terminus of the oligonucleotide through amido bond, and captured the products of HCR to form triplex DNA. In this case, the AIE fluorophores were located in close proximity to generate fluorescence. This assay provided turn-on fluorescence efficiency with a high signal-to-noise ratio and excellent amplification capability to solve the shortcoming of HCR-based fluorescence sensing methods. It enabled sensitive detection of Vibrio parahaemolyticus in the range of 102-106 CFU mL-1, and with a low limit of detection down to 39 CFU mL-1. In addition, this assay expressed good specificity and practicability. The triplex DNA-based AIE probe forms a universal molecular tool for developing HCR-based fluorescence sensing methods.


Assuntos
Técnicas Biossensoriais , DNA , DNA/genética , DNA/química , Hibridização de Ácido Nucleico/métodos , Corantes Fluorescentes/química , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas Biossensoriais/métodos , Limite de Detecção
19.
Oncol Lett ; 27(5): 221, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38586211

RESUMO

MicroRNAs (miRNAs) were previously demonstrated to be involved in the pathogenesis of non-small-cell lung cancer (NSCLC); however, the roles of certain miRNAs in NSCLC remain to be elucidated. The present study aimed to investigate the functions of screened miRNAs in NSCLC and the potential mechanisms. First, expression profiles of miRNAs were downloaded from the Gene Expression Omnibus (dataset no. GSE29248) and the differentially expressed miRNAs were analyzed by bioinformatics methods. Reverse transcription-quantitative PCR was used to validate the differential expression of miR-373 in clinical samples. The association between miR-373 expression levels and clinicopathological characteristics was also investigated. To further examine how miR-373 mediates the emergence of NSCLC, western blot, Cell Counting Kit-8, cell invasion and wound-healing assays, as well as apoptosis detection and a luciferase assay were used. The results indicated significant downregulation of miR-373 in NSCLC tissues and its low expression was closely associated with the degree of differentiation, clinical stage and tumor size, and was indicative of an unfavorable prognosis for patients with NSCLC. A functional study indicated that overexpression of miR-373 inhibited the proliferation, promoted apoptosis, and suppressed invasion and migration of NSCLC cells. Bioinformatics prediction and functional assays suggested that Grb-associated binding protein 2 (GAB2) was a direct target of miR-373. In addition, GAB2 was found to be significantly upregulated in NSCLC tissues, and clinically, miR-373 was negatively associated with GAB2. Furthermore, overexpression of GAB2 blocked the tumor suppressive effects of miR-373 on NSCLC cells. Mechanistically, miR-373 mimics were able to reduce the expression of GAB2 and subsequently decrease the phosphorylation level of AKT and mTOR protein. The present results indicate that miR-373 exerts its anti-tumor effects in NSCLC cells by targeting the GAB2/PI3K/AKT pathway, suggesting that miR-373 may be a potential therapeutic target in NSCLC.

20.
Photochem Photobiol ; 2024 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-39080821

RESUMO

In recent years, studies have shown that low-dose supplemental infrared (IR) irradiation exhibits systemic anti-inflammatory effects. The gut microbiota is increasingly recognized as a potential mediator of these effects due to its role in regulating host metabolism and inflammatory responses. To investigate the role of gut microbiota diversity and metabolite changes in the mechanism of light-emitting diodes (LED) infrared's anti-inflammatory action, we conducted IR irradiation on mice. Serum inflammatory cytokines were measured using ELISA, and fecal samples were subjected to metagenomic, untargeted, and targeted metabolomic analyses. Our results demonstrated a significant increase in the anti-inflammatory cytokine IL-10 in the IR group, accompanied by a declining trend in pro-inflammatory cytokines. Gut microbiome analysis revealed distinct alterations in composition and functional genes between the groups, including the enrichment of beneficial bacteria like various species of Parabacteroides and Akkermansia muciniphila in the IR group. Notably, the IR group exhibited enrichment in carbohydrate metabolism pathways and a reduction in DNA damage and repair pathways. Furthermore, targeted metabolomic analysis highlighted a notable increase in short-chain fatty acids (SCFAs), including butyric acid and isobutyric acid, which positively correlated with the abundance of several beneficial bacteria. These findings suggest a potential interplay between gut microbiota-derived SCFAs and the anti-inflammatory response. In conclusion, our study provides comprehensive insights into the changes in gut microbiota species and functions associated with IR irradiation. Moreover, we emphasize the significance of altered SCFAs levels in the IR group, which may contribute to the observed anti-inflammatory effects. Our findings contribute valuable evidence supporting the role of low-dose infrared light irradiation as an anti-inflammatory therapy.

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