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1.
Proc Natl Acad Sci U S A ; 121(25): e2316551121, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38865260

RESUMO

The NLRP3 inflammasome, a pivotal component of innate immunity, has been implicated in various inflammatory disorders. The ubiquitin-editing enzyme A20 is well known to regulate inflammation and maintain homeostasis. However, the precise molecular mechanisms by which A20 modulates the NLRP3 inflammasome remain poorly understood. Here, our study revealed that macrophages deficient in A20 exhibit increased protein abundance and elevated mRNA level of NIMA-related kinase 7 (NEK7). Importantly, A20 directly binds with NEK7, mediating its K48-linked ubiquitination, thereby targeting NEK7 for proteasomal degradation. Our results demonstrate that A20 enhances the ubiquitination of NEK7 at K189 and K293 ubiquitinated sites, with K189 playing a crucial role in the binding of NEK7 to A20, albeit not significantly influencing the interaction between NEK7 and NLRP3. Furthermore, A20 disrupts the association of NEK7 with the NLRP3 complex, potentially through the OTU domain and/or synergistic effect of ZnF4 and ZnF7 motifs. Significantly, NEK7 deletion markedly attenuates the activation of the NLRP3 inflammasome in A20-deficient conditions, both in vitro and in vivo. This study uncovers a mechanism by which A20 inhibits the NLRP3 inflammasome.


Assuntos
Inflamassomos , Quinases Relacionadas a NIMA , Proteína 3 que Contém Domínio de Pirina da Família NLR , Proteína 3 Induzida por Fator de Necrose Tumoral alfa , Ubiquitinação , Quinases Relacionadas a NIMA/metabolismo , Quinases Relacionadas a NIMA/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Inflamassomos/metabolismo , Animais , Camundongos , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/metabolismo , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/genética , Humanos , Macrófagos/metabolismo , Macrófagos/imunologia , Células HEK293 , Camundongos Knockout , Ligação Proteica
2.
Anal Bioanal Chem ; 415(23): 5755-5767, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37540345

RESUMO

Novel hydrophilic poly(N, N-methylenebisacrylamide/1,2-epoxy-5-hexene) coated magnetic nanospheres functionalized with 2-aminopurine (denoted as Fe3O4@poly(MBA/EH)@2AP) for enriching glycopeptides and glycosylated exosomes were successfully obtained using a simple and green method on the basis of the HILIC (hydrophilic interaction liquid chromatography) enrichment strategy. The high density of polar groups endows the material with amazing hydrophilicity, enabling the nanomaterial to successfully capture glycopeptides and glycosylated exosomes within 1 min. Meanwhile, the materials demonstrated great sensitivity (0.01 fmol/µL), good loading capability (125 µg/mg), high selectivity (BSA:HRP = 1000:1), and repeatability (more than 10 times). Besides, the material was applied in the analysis of bio-samples, a total of 290 glycosylated peptides and 184 glycosylation sites mapping to 185 glycoproteins were identified in the serum of uremic patients. Besides, 42 glycopeptides were enriched from the saliva of healthy people. At the same time, it was verified by TEM and western blot that the complete glycosylated exosomes were successfully captured from the serum of the uremic patients. All experiments have demonstrated that Fe3O4@poly(MBA/EH)@2AP has a promising future in practical applications.


Assuntos
Exossomos , Nanoestruturas , Humanos , Glicopeptídeos/química , Glicosilação , Polímeros , Exossomos/química , Interações Hidrofóbicas e Hidrofílicas , Fenômenos Magnéticos
3.
Oral Dis ; 29(8): 3447-3459, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35957556

RESUMO

OBJECTIVES: Increasing evidence indicated circRNAs were involved in stem cells osteogenesis differentiation. Herein, we aimed to clarify the role of hsa-circ-0107593 during the osteogenesis process of human adipose-derived stem cells (hADSCs) and the underlying mechanisms. METHODS: The ring structure of hsa-circ-0107593 was confirmed using RNase R treatment and Sanger sequencing. Nucleoplasmic separation and fluorescence in situ hybridization detected hsa-circ-0107593 distribution. Lentivirus and siRNA were used to modulate the expression of hsa-circ-0107593, and the binding relationship between hsa-circ-0107593 and miR-20a-5p was verified by luciferase assay and RNA immunoprecipitation. We detected the osteogenic activity of hADSCs through alkaline phosphatase staining, alizarin red S staining, real-time polymerase chain reaction (RT-PCR), western blot, and cellular immunofluorescence experiment. In vivo, micro-computed tomography was performed to analyze bone formation around skull defect. RESULTS: RT-PCR results exhibited that hsa-circ-0107593 was downregulated while miR-20a-5p was upregulated during hADSCs osteogenesis. In vivo and in vitro experiments results indicated that knocking down hsa-circ-0107593 promoted the osteogenic differentiation of hADSCs, while overexpression of hsa-circ-0107593 showed an inhibitory effect on hADSCs osteogenic differentiation. In vitro experiment results showed hsa-circ-0107593 acted as a hADSCs osteogenic differentiation negative factor for it inhibited the suppressing effect of miR-20a-5p on SMAD6. CONCLUSION: Knocking down hsa-circ-0107593 acts as a positive factor of the osteogenic differentiation of hADSCs via miR-20a-5p/SMAD6 signaling.


Assuntos
MicroRNAs , Osteogênese , Humanos , Osteogênese/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Regulação para Baixo , Hibridização in Situ Fluorescente , Microtomografia por Raio-X , Diferenciação Celular/genética , Proliferação de Células/genética , Proteína Smad6/genética , Proteína Smad6/metabolismo
4.
J Cell Physiol ; 237(2): 1225-1244, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34796958

RESUMO

Circular RNA (circRNA) is a class of endogenous noncoding RNA (ncRNA), presenting as a special covalent closed loop without a 5' cap or 3' tail, maintaining resistance to RNA exonuclease and keeping high stability. Although lowly expressed in most situations, circRNA makes an active difference in regulating physiological or pathological processes by modulating gene expression by regulation of transcription, protein, and miRNA functions through various mechanisms in particular tissues. Recent studies have demonstrated the roles of the miRNA-circRNA network in the development of several bone diseases such as osteoporosis, a multiple-mechanism disease resulting from defective bone quality and low bone mass, osteoarthritis, whose main pathomechanism is inflammation and articular cartilage degradation, as well as osteosarcoma, known as one of the most common bone cancers. However, the specific mechanism of how circRNA along with miRNA influences those diseases is not well documented, showing potential for the development of new therapies for those bone diseases.


Assuntos
Neoplasias Ósseas , Cartilagem Articular , MicroRNAs , Osteoartrite , Osteossarcoma , Neoplasias Ósseas/metabolismo , Cartilagem Articular/metabolismo , Redes Reguladoras de Genes , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Osteoartrite/genética , Osteoartrite/metabolismo , Osteossarcoma/genética , Osteossarcoma/metabolismo , RNA Circular/genética , RNA não Traduzido/metabolismo
5.
Pharmacol Res ; 166: 105497, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33609697

RESUMO

Osteoarthritis (OA) is a major cause of disability in the elderly population and represents a significant public health problem and socioeconomic burden worldwide. However, no disease-modifying therapeutics are currently available for OA due to an insufficient understanding of the pathogenesis of this disability. As a unique cell type in cartilage, chondrocytes are essential for cartilage homeostasis and play a critical role in OA pathogenesis. Mitochondria are important metabolic centers in chondrocytes and contribute to cell survival, and mitochondrial quality control (MQC) is an emerging mechanism for maintaining cell homeostasis. An increasing number of recent studies have demonstrated that dysregulation of the key processes of chondrocyte MQC, which involve mitochondrial redox, biogenesis, dynamics, and mitophagy, is associated with OA pathogenesis and can be regulated by the chondroprotective molecules 5' adenosine monophosphate-activated protein kinase (AMPK) and sirtuin 3 (SIRT3). Moreover, AMPK and SIRT3 regulate each other, and their expression and activity are always consistent in chondrocytes, which suggests the existence of an AMPK-SIRT3 positive feedback loop (PFL). Although the precise mechanisms are not fully elucidated and need further validation, the current literature indicates that this AMPK-SIRT3 PFL regulates OA development and progression, at least partially by mediating chondrocyte MQC. Therefore, understanding the mechanisms of AMPK-SIRT3 PFL-mediated chondrocyte MQC in OA pathogenesis might yield new ideas and potential targets for subsequent research on the OA pathomechanism and therapeutics.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Condrócitos/patologia , Osteoartrite/patologia , Transdução de Sinais , Sirtuína 3/metabolismo , Animais , Condrócitos/metabolismo , Progressão da Doença , Humanos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Mitofagia , Osteoartrite/metabolismo
6.
J Cell Physiol ; 235(7-8): 5490-5500, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32030759

RESUMO

Tumor microenvironment is known to play important roles in tumor progression. Many therapies, targeting the tumor microenvironment, are designed and applied in the clinic. One of these approaches is in situ antitumor therapy. This way, bacteria, antibodies, plasmid DNA, viruses, and cells are intratumorally delivered into the tumor site as "in-situ antitumor vaccine," which seeks to enhance immunogenicity and generate systemic T cell responses. In addition, this intratumoral therapy can alter the tumor microenvironment from immunosuppressive to immunostimulatory while limiting the risk of systemic exposure and associated toxicity. Contemporarily, promising preclinical results and some initial success in clinical trials have been obtained after intratumoral therapy.


Assuntos
Vacinas Anticâncer/uso terapêutico , Imunoterapia , Neoplasias/terapia , Microambiente Tumoral/imunologia , Vacinas Anticâncer/imunologia , Terapia Combinada , Humanos , Imunização , Neoplasias/imunologia , Neoplasias/patologia , Linfócitos T/imunologia
7.
Ren Fail ; 40(1): 666-671, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30489209

RESUMO

OBJECTIVE: Uric acid (UA) is a risk marker of CKD and SUA level in CKD 3-4 patients closely correlates with hyperuricemic nephropathy (HN) morbidity. This study was designed to evaluate the risk factors for HN in CKD 3-4 patients. METHODS: The 461 CKD 3-4 patients were recruited and all patients were divided into three groups (24 h UUA normal, underexeret, and overproduct type groups) according to the 24 h UUA level after receiving low purine food for five days. Clinical and biochemical characteristics of CKD patients were collected for the logistic regression analysis. Correlation analysis of the mRNA relative expression level of hUAT and hURAT1 with serum UA (SUA) level also was evaluated. RESULTS: There were significant increases in characteristics including average age, waist-to-height ratio (WHR), SUA levels, HN ratio, TG/HDL ratio, body mass index (BMI), blood pressure (BP), uNgal/Cr. ratio, and uKim-1/Cr. ratio in overproduct type group in comparison with the other two groups. Logistic regression analysis showed SUA, CHO, uKim-1/Cr. ratio and uNgal/Cr. ratio were independent and multiple risk factors for HN. Moreover, hUAT and hURAT1 mRNA relative expression levels were significantly correlated with SUA level in the underexeret type CKD 3-4 patients. CONCLUSIONS: These results showed SUA and other characteristics contributed to HN morbidity in CKD 3-4 patients.


Assuntos
Hiperuricemia/epidemiologia , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/epidemiologia , Ácido Úrico/sangue , Adulto , Idoso , Pressão Sanguínea , Índice de Massa Corporal , China/epidemiologia , Estudos Epidemiológicos , Feminino , Humanos , Hiperuricemia/sangue , Masculino , Pessoa de Meia-Idade , Transportadores de Ânions Orgânicos/genética , Proteínas de Transporte de Cátions Orgânicos/genética , RNA Mensageiro/análise , Análise de Regressão , Fatores de Risco , Fatores Sexuais
8.
J Sci Food Agric ; 97(7): 2199-2204, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27611871

RESUMO

BACKGROUND: Mannanases are important enzymes which are widely used as a tool in agriculture and food industries. To improve the performance of mannanase Man23, a mutant library was created with rational design, and mutations were introduced on loops around the catalytic region. The Brevibacillus brevis B16 cell-free system which was created in this experiment provided the ability to express the mutant library efficiently. The activities of mutants were measured with a multi-volume spectrophotometer. RESULTS: The mutant Man1606 gained from this system is a sextet which has mutations of N146G, S147H, S156P, T157Y, Q206S and T249H simultaneously on loops 6, 8 and 10. Man1606 showed higher activity and stability than Man23. The optimal temperature of Man1606 rose by 5 °C (from 55 to 60 °C) and the optimal pH increased slightly but its range became broader. CONCLUSION: This experiment demonstrated the B. brevis cell-free system shortens the expression time and is an efficient tool for mannanase engineering. © 2016 Society of Chemical Industry.


Assuntos
Bacillus subtilis/enzimologia , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , beta-Manosidase/química , beta-Manosidase/genética , Bacillus subtilis/química , Bacillus subtilis/genética , Proteínas de Bactérias/metabolismo , Clonagem Molecular , Estabilidade Enzimática , Expressão Gênica , Biblioteca Gênica , Concentração de Íons de Hidrogênio , Cinética , Temperatura , beta-Manosidase/metabolismo
9.
J Sci Food Agric ; 96(4): 1390-5, 2016 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-25930671

RESUMO

BACKGROUND: The disordered residues on distal loops affect the molecular structural stability and on some occasions have regulatory roles in catalytic reaction. To increase understanding of the influence of distal residue mutation, this study explored the thermostability and enzymatic activity of mannanase Man1312 deletion mutants. The focus was on residues located on the N-terminal region because they are more disordered and changeable. The effects of N-terminal truncation on enzymatic activity and thermal dynamics were investigated by spectrophotometry, circular dichroism and differential scanning calorimetry assays. RESULTS: The deletion mutants on V3, N7 and Q11 showed a marked increase in stability, while the enzymatic activity was significantly improved when triplet deletion was carried out. Triplet deletion MandVNQ showed around double the stability of its corresponding single-site and double-site deletion mutants. The Tm value of MandVNP was about 8 °C higher than that of Man1312. MandVNP had improved characteristics of Topt by 10 °C, t1/2 by 10 min and catalytic activity by 11% in comparison with Man1312. Analysis of spectra and modeling showed that MandVNQ had increased helix and strand contents. CONCLUSION: N-terminal truncation had positive effects on the thermostability and activity of mannanase.


Assuntos
Estabilidade Enzimática , Temperatura Alta , beta-Manosidase/química , beta-Manosidase/metabolismo , Sequência de Aminoácidos , Modelos Moleculares , Estrutura Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/fisiologia , Estrutura Terciária de Proteína , Deleção de Sequência , Relação Estrutura-Atividade , beta-Manosidase/genética
10.
BMC Psychiatry ; 14: 125, 2014 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-24774860

RESUMO

BACKGROUND: Depression is the most widely acknowledged psychological problem among end-stage renal disease (ESRD) patients. Depression may be associated with VD deficiency. The aims of this study are to (a) elucidate the prospective association between HsCRP, VD contents and depressive symptoms in the dialyzed population, and (b) find the effect of calcitriol supplementation on depression in dialyzed patients. METHODS: In this prospective study, 484 dialysis patients (382 hemodialysis [HD] cases and 102 peritoneal dialysis [PD] cases; aged 18-60 years) from two hospitals in southeast China were included. The depression in these patients was evaluated using the Chinese version of Beck's Depression Inventory (BDI). All subjects answered the BDI-I questionnaire for assessment of depression levels in summer. A cut-off value of 16 was set to include dialysis patients with depression. All patients were divided into two groups depending on the absence (Group 1) or presence (Group 2) of depression. The two groups took 0.5 µg/day 1,25-Dihydroxyvitamin D orally for one year. BDI Scores were recalculated for all patients. Sociodemographic, clinical data, and serum VD contents were also collected. RESULTS: A total of 484 participants (247 men [51.0%] and 237 women [49.0%]) were surveyed. Depressive symptoms were found in 213 (44.0%) patients. The baseline serum VD level (VD2 + VD3) was 17.6 ± 7.7 nmol/L. Patients with depressive symptoms have significantly higher serum HsCRP level and significantly lower serum VD level compared with the control group. After one-year follow-up, the supplementation of 0.5 µg/day calcitriol slightly improved the microinflammatory state such as lowering mean serum HsCRP level and improving serum VD level, but not in significantly enhancing the depressive symptoms. CONCLUSIONS: Calcitriol supplementation did not significantly enhance the depressive symptoms in our dialyzed population although patients with low levels of serum VD were more depressed. Therefore, more prospective randomized controlled trials are necessary to reveal the exact cause-and-effect relationship between VD status and depressive symptoms or VD status related to some specific subtypes in dialyzed patients.


Assuntos
Depressão/sangue , Transtorno Depressivo/sangue , Falência Renal Crônica/psicologia , Vitamina D/análogos & derivados , Vitaminas/sangue , Adolescente , Adulto , Calcitriol/administração & dosagem , China , Suplementos Nutricionais , Feminino , Humanos , Falência Renal Crônica/sangue , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Diálise Peritoneal , Estudos Prospectivos , Escalas de Graduação Psiquiátrica , Diálise Renal , Inquéritos e Questionários , Vitamina D/sangue , Adulto Jovem
11.
BMC Complement Med Ther ; 24(1): 194, 2024 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-38760722

RESUMO

OBJECTIVE: To study the effect of mindfulness meditation combined with progressive muscle relaxation training on the clinical efficacy and quality of life in patients with sarcopenia receiving maintenance haemodialysis (MHD). METHODS: Eligible patients with sarcopenia in our hospital were randomly assigned to a control group (n = 24) and an intervention group (n = 25). The control group received conventional dialysis treatment, while the intervention group underwent mindfulness meditation combined with progressive muscle relaxation training during the interdialysis period in addition to conventional dialysis treatment. The effect of the intervention was evaluated after 12 weeks. RESULTS: There were no significant differences in the baseline values of various parameters between the two groups. Exercise capacity (sit-to-stand test,handgrip,time to 10 sit-ups) significantly improved in the intervention group after 12 weeks (32.68 ± 8.32 vs 26.50 ± 6.83; 37.42 ± 10.12 vs 28.12 ± 8.51; 19.8 ± 5.40 vs 25.29 ± 7.18) (p < 0.05). In terms of the kidney disease quality of life (KDQOLTM) score, all other dimensions except sexual function, social functioning, burden of kidney disease and work status dimensions showed significant improvement compared to the baseline (p < 0.05). In the control group, only the dialysis staff encouragement (DSE) and patient satisfaction (PS) dimensions showed slight improvements compared to the baseline (p > 0.05). When compared with the control group, the intervention group showed significant improvements in 10 dimensions of exercise capacity and KDQOLTM scores for physical function, role-physical, general health, energy, symptom/problem list, sleep, DSE, pain, cognitive function, emotional well-being and patient PS after 12 weeks (61.30 ± 5.38 vs 42.98 ± 5.73; 57.50 ± 3.55 vs 50.70 ± 3.62) (p < 0.05). Some inflammatory markers, such as the levels of interleukin-6 and high-sensitivity C-reactive protein (30.29 ± 2.96 vs 17.65 ± 3.22; 8.93 ± 0.99 vs 3.02 ± 0.34), showed a decrease during the intervention, while albumin and prealbumin levels were significantly increased compared with the baseline (30.62 ± 1.65 vs 35.60 ± 1.68; 0.32 ± 0.05 vs 0.44 ± 0.07) (p < 0.05). CONCLUSION: Combined intervention training can improve the motor ability and quality of life of patients with sarcopenia within a short period of time.


Assuntos
Meditação , Atenção Plena , Qualidade de Vida , Diálise Renal , Sarcopenia , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Atenção Plena/métodos , Idoso , Sarcopenia/terapia , Resultado do Tratamento
12.
J Multidiscip Healthc ; 17: 3743-3751, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39104383

RESUMO

Objective: Sarcopenia is more common in maintenance hemodialysis (MHD) patients, and the aim of this study is to analyze the risk factors associated with sarcopenia in MHD patients, along with its correlation to emotional status and quality of life. Methods: This is a cross-sectional cohort study. A total of 111 MHD patients who were treated in the Department of Nephrology of our hospital were selected as the study subjects by convenience sampling. The quality of life and emotional status were evaluated by health survey scale (SF-36), self-rating anxiety scale (SAS) and self-rating depression scale (SDS). Regression analysis was used to explore the influencing factors of sarcopenia. Correlation analysis was used to investigate the correlation between sarcopenia and quality of life and emotional status. Results: The prevalence of sarcopenia was 59.8%. The results showed that age, gender, body mass index (BMI), dialysis time, economic status, marital status and pre-dialysis creatinine were significant factors affecting the development of sarcopenia in hemodialysis patients (p<0.05). The SF-36 total score was significantly lower in the sarcopenia group (72.05±12.28 vs 78.03±10.55) than in the non-sarcopenia group, but the anxiety scale score (52.97±4.67 vs 36.2±3.36) and depression scale score (57.67±4.58 vs 38.71±3.77) were significantly higher than those in the non-sarcopenia group (p< 0.001). Correlation analysis showed that sarcopenia was positively correlated with SAS and SDS scores and negatively correlated with SF-36 total score (p < 0.05). Conclusion: The risk of sarcopenia was higher among MHD patients who were older, male, single, with a longer MHD duration, lower economic status, lower BMI, comorbid diabetes and lower levels of creatinine.

13.
Microb Cell Fact ; 12: 78, 2013 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-24011469

RESUMO

BACKGROUND: Mannanase is an enzyme that can catalyze random hydrolysis of beta-1,4-mannosidic linkages in the main chain of mannans, glucomannans and galactomannans which are the key polymers in hemicellulose. It has been used in a number of different industrial applications including food, feed, pharmaceutical, pulp/paper industries, and second generation biofuel. To optimize the expression system of mannanase Man23 gene, two kinds of vectors and host bacteria were determined and compared. RESULTS: Recombinants pHY-p43-man23 and pBPS-man23 were constructed and transferred into Bacillus subtilis WB600 and Brevibacillus brevis respectively. For mannanase Man23 gene, recombinant pHY-p43-man23 expressed in Brevibacillus brevis had higher production and activity. Compared to the wild-type Bacillus subtilis B23, the production of recombinant pHY-p43-man23 in B. brevis increased by 10 times and activity increased by 21.3%. pHY-p43-man23 in B. brevis had activity at the range of 20 ~ 70 °C but its optimum temperature was 50 °C and had activity from pH 4 ~ 10 but its optimum pH was around 7. This demonstrated the recombinant had improved stability as well. CONCLUSIONS: Mannanase is an important industrial enzyme and combination of vector pHY-p43 and host Brevibacillus brevis is a novel expression system for a mannanase decoding gene. This work aims at exploring a better expression system of mannanase Man23 decoding gene for industrial application.


Assuntos
Bacillus subtilis/metabolismo , beta-Manosidase/biossíntese , Sequência de Aminoácidos , Bacillus subtilis/enzimologia , Bacillus subtilis/genética , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , beta-Manosidase/química , beta-Manosidase/genética
14.
Anal Methods ; 15(21): 2677-2684, 2023 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-37226592

RESUMO

In this study, we developed a green, one-step hydrothermal carbonization (HTC) method that used water as the sole solvent to create boronic acid group-rich carbonaceous spheres (BCS). When the abundant boronic acid groups on the carbonaceous spheres react with the hydroxyl groups on the glycans in an alkaline environment, the glycopeptides are specifically captured. The results showed that BCS had excellent detection limits (0.1 fmol µL-1), selectivity (1 : 1000), and stability (10 cycles). In addition, the BCS also demonstrated excellent glycopeptide enrichment capabilities in complex biological samples; 219 glycopeptides attributed to 167 glycoproteins and 235 glycopeptides related to 166 glycoproteins in PE patient and normal pregnancy control serum were identified by nano LC-MS/MS, respectively. In addition, the molecular function of heparin binding and biological process of complement activation, positive regulation of immune response, and positive regulation of tumor necrosis factor production were significantly different between PE patients and healthy pregnant women according to gene ontology analysis, indicating that these may be associated with the development of PE.


Assuntos
Pré-Eclâmpsia , Espectrometria de Massas em Tandem , Gravidez , Humanos , Feminino , Glicopeptídeos/análise , Glicopeptídeos/química , Ácidos Borônicos/química , Glicoproteínas/química , Glicoproteínas/metabolismo
15.
Arthritis Res Ther ; 22(1): 220, 2020 09 21.
Artigo em Inglês | MEDLINE | ID: mdl-32958016

RESUMO

A20, also known as TNF-α-induced protein 3 (TNFAIP3), is an anti-inflammatory protein that plays an important part in both immune responses and cell death. Impaired A20 function is associated with several human inflammatory and autoimmune diseases. Although the role of A20 in mediating inflammation has been frequently discussed, its intrinsic link to arthritis awaits further explanation. Here, we review new findings that further demonstrate the molecular mechanisms through which A20 regulates inflammatory arthritis, and we discuss the regulation of A20 by many factors. We conclude by reviewing the latest A20-associated mouse models that have been applied in related research because they reflect the characteristics of arthritis, the study of which will hopefully cast new light on anti-arthritis treatments.


Assuntos
Artrite , Doenças Autoimunes , Anti-Inflamatórios , Humanos , Inflamação , NF-kappa B , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa
16.
Curr Gene Ther ; 20(2): 152-161, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32951573

RESUMO

BACKGROUND: Osteoporosis is the most common metabolic bone disease. There is still an unmet need for novel therapeutic agents that could be beneficial as osteoporosis treatments. It has been reported that the neurotransmitter γ-aminobutyric acid (GABA) might be associated with human bone formation. However, the precise mechanism remains unclear. OBJECTIVE: To investigate the effect of GABA on bone metabolism and explore the possible role of TNFAIP3 in this process. METHODS: GABA had little effect on the proliferation of human mesenchymal stem cells (hMSCs) and RAW 264.7 cells, as indicated by the cell counting kit-8 (CCK-8) assay. The results showed that GABA enhanced the intensity of ALP staining, ALP activity, and accumulation of Ca2+ mineralized nodules in hMSCs during osteogenic induction. RESULTS: The qRT-PCR results indicated that GABA treatment significantly increased the mRNA expression of osteogenic genes in hMSCs. In RAW 264.7 cells, TRAP staining showed that GABA did not alter the number or size of osteoclasts or the expression of osteoclastic genes, which suggests that GABA does not affect osteoclastic differentiation. Mechanistically, GABA treatment significantly induced the sustained expression of TNFAIP3. Furthermore, by knocking down TNFAIP3, the osteogenic effect of GABA was antagonized, which suggests that TNFAIP3 mediates the effects of GABA in hMSCs. CONCLUSION: Our results suggested that GABA treatment positively regulated osteogenic differentiation by upregulating TNFAIP3, while no obvious effect on osteoclastic differentiation was detected. Therefore, our results provide a potential gene therapy for the treatment of osteoporosis and low bone mineral density.


Assuntos
Doenças Ósseas Metabólicas/tratamento farmacológico , Osteoporose/tratamento farmacológico , Proteína 3 Induzida por Fator de Necrose Tumoral alfa/genética , Ácido gama-Aminobutírico/farmacologia , Animais , Doenças Ósseas Metabólicas/genética , Doenças Ósseas Metabólicas/metabolismo , Doenças Ósseas Metabólicas/patologia , Osso e Ossos/metabolismo , Proliferação de Células/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Osteogênese/efeitos dos fármacos , Osteoporose/genética , Osteoporose/metabolismo , Osteoporose/patologia , Células RAW 264.7
17.
Yi Chuan ; 28(8): 1051-4, 2006 Aug.
Artigo em Zh | MEDLINE | ID: mdl-16870596

RESUMO

Trans-translation is a mechanism that fixes stalled translation course. tmRNA is the core of trans-translation and it has dual function as a tRNA and a mRNA. tmRNA recognizes the ribosome carrying truncated mRNA with specificity under the help SmpB protein. It can be directed to A site by ribosomal protein S1. At first tmRNA prolongs the genetic message on the stalled mRNA, then termination codon stops peptide synthesis to form a nascent chain with the tag sequence. Finally, tmRNA-SmpB system frees stalled ribosomes and directs degradation of the products of these frustrated protein synthesis reactions. This paper introduces the most recent studies on trans-translation.


Assuntos
Escherichia coli/genética , Biossíntese de Proteínas , Escherichia coli/química , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Modelos Genéticos , RNA Mensageiro/química , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA de Transferência/genética , RNA de Transferência/metabolismo , Ribossomos/química , Ribossomos/genética , Ribossomos/metabolismo
18.
Appl Biochem Biotechnol ; 180(1): 122-35, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27193255

RESUMO

Hemicelluloses are the second major polysaccharides in nature and can be converted to ethanol product by a variety of enzymes including mannanases. Mannanase is an important enzyme that hydrolyses mannose-containing polysaccharides which are abundant in plants. An optimized mannanase could help to improve conversion process and make the technology efficiently and competitively. In this work, the effects of loops adjacent to active region on enzymic properties of Man1312 were investigated. Loop 6 and 10 are two loops neighboring to Man1312 catalytic region, and deletion mutagenesis and residue substitution were performed on both loops. Deletion on sites S145, Q148, N244, and S255 and substitution on sites N146, S147, S156, and T157 gave significant increased stability to enzyme. The quadruplet mutant ManD4I4 combined all the mutations and had higher optimal temperature and T m value by 5 and 4 °C than Man1312, respectively. From our data, we are able to conclude the loops of enzymes are important to design mutagenesis and obtain improved properties, especially the loops neighboring to catalytic region from tertiary structure. In our experiment, residue deletion and substitution on loops neighboring to catalytic region made significant improvement on enzyme properties.


Assuntos
Bacillus subtilis/enzimologia , beta-Manosidase/química , beta-Manosidase/metabolismo , Substituição de Aminoácidos , Varredura Diferencial de Calorimetria , Domínio Catalítico , Dicroísmo Circular , Estabilidade Enzimática , Ligação de Hidrogênio , Concentração de Íons de Hidrogênio , Cinética , Mutagênese , Proteínas Mutantes/química , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Deleção de Sequência , Relação Estrutura-Atividade , Temperatura
19.
J Nanosci Nanotechnol ; 4(5): 553-7, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15503442

RESUMO

A novel chemically modified electrode has been prepared on the basis of the attachment of multi-wall carbon nanotubes (MWNT) onto the surface of a glassy carbon electrode (GCE) in the presence of a hydrophobic surfactant, dihexadecyl phosphate (DHP). This MWNT film was characterized by transmission electron microscopy images (TEM) and scanning electron microscopy (SEM). The electrochemical behavior of tyrosine at the MWNT film coated GCE was examined and it is found that this MWNT-modified GCE greatly enhances the oxidation peak current of tyrosine. Effects of some important factors, including pH, scan rate and amount of modifier, on the oxidation process of tyrosine were investigated. When the signal to noise ratio (SNR) is 3, the detection limit is 1 x 10(-7) M. The low relative standard deviations of the detection of tyrosine in human morning urine (3.3%) and white wine (5.2%) suggest a good reproducibility of the modified electrode.


Assuntos
Materiais Revestidos Biocompatíveis/química , Cristalização/métodos , Eletroquímica/instrumentação , Eletrodos , Nanotubos de Carbono/química , Nanotubos de Carbono/ultraestrutura , Tirosina/análise , Tirosina/química , Adsorção , Carbono/química , Eletroquímica/métodos , Teste de Materiais , Propriedades de Superfície
20.
J Int Med Res ; 42(3): 718-27, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24781720

RESUMO

OBJECTIVE: To investigate the effect of individualized exercise on exercise capacity and health-related quality of life (HRQoL) in uraemic patients during maintenance haemodialysis (MHD). METHODS: Patients receiving MHD were divided randomly into a test group, who underwent recumbent cycling exercise during dialysis, and a control group, who performed simple stretching exercises. The same dialysis protocol was used for all study participants. At study start and after 12 weeks, exercise capacity was measured using tests of physical ability; HRQoL was measured using the kidney disease quality of life score (KDQOL-SF™). RESULTS: A total of 65 patients were included in the study: 33 in the control group and 32 in the test group. There were no significant differences in patient characteristics between the two groups at baseline. After 12 weeks, there were significant improvements in exercise capacity and in many of the items of the KDQOL-SF™ in the test group compared with the control group. CONCLUSION: Individualized exercise during MHD significantly improved the exercise capacity and HRQoL for uraemic patients within a short time period, and could therefore be used as a simple, cost-effective therapeutic approach.


Assuntos
Terapia por Exercício/métodos , Qualidade de Vida/psicologia , Diálise Renal , Insuficiência Renal Crônica/terapia , Uremia/terapia , Adulto , Técnicas de Exercício e de Movimento , Teste de Esforço , Feminino , Força da Mão , Humanos , Testes de Função Renal , Masculino , Pessoa de Meia-Idade , Pacientes Ambulatoriais , Medicina de Precisão , Insuficiência Renal Crônica/fisiopatologia , Insuficiência Renal Crônica/psicologia , Uremia/fisiopatologia , Uremia/psicologia
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