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1.
Complement Ther Clin Pract ; 53: 101797, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37690375

RESUMO

PURPOSE: This study aims to develop and validate a concise tool for evaluating acupuncture expectancy that is easy to understand and conforms to acupuncture characteristics. MATERIALS AND METHODS: A draft was created using the Delphi consensus method. Reliability, validity, discrimination, and feasibility tests were conducted at the item and scale levels. RESULTS: The scale themes were defined as disease-related, treatment-related, process-related, and outcome-related. After two rounds of Delphi surveys with good experts' reliability (authority coefficients of experts were 0.86 and 0.87 in the two rounds) and agreement (Kendall's concordance coefficient of the participants were 0.33 and 0.15 in the two rounds, P < 0.05), 11 items (the mean score for item importance, full mark ratios, and coefficient of variation of items were ≥3.5, ≥25%, and ≤0.30, respectively) were included in the draft. A total of 145 individuals were recruited to test the draft. Reliability was assessed by Cronbach's α coefficient (0.90), split-half reliability coefficient (0.89), and test-retest reliability (Pearson's coefficient = 0.74, P < 0.05). Content validity was assessed by the content validity index (Item-CVI ≥ 0.78 and Scale-CVI/Ave = 0.92), and a confirmatory factor analysis was performed to assess the construct validity. The discrimination of scale items was evaluated by the critical ratio (CR > 3.00) and the homogeneity test (item-total correlations >0.40). Feasibility was assessed through the acceptance rate (recovery rate = 98.60%, response rate = 100%), completion rate (100%), and completion time (4.99 ± 6.80 min). CONCLUSION: The patients' expectancy scale of acupuncture (PESA) consists of 11 items with four themes, disease-related, treatment-related, process-related, and outcome-related. It has great reliability, validity, discrimination, and feasibility and has the potential to evaluate acupuncture expectancy in clinical trials.


Assuntos
Terapia por Acupuntura , Humanos , Reprodutibilidade dos Testes , Psicometria/métodos , Inquéritos e Questionários , Análise Fatorial
2.
Anal Chim Acta ; 1276: 341609, 2023 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-37573105

RESUMO

Compared with antibody, the recognition spectrum of a receptor is broader, and its recognition ability can be improved using simple mutagenesis technique. Compared with conventional immunoassay, the magnetic bead based immunoassay is simpler and can be recycled. Compared with colorimetric and luminescent immunoassays, fluoroimmunoassay is simpler because it does not require a substrate. So a method combines these merits is desirable. In this study, two amino acids in the binding pocket of a natural Escherichia coli TetR protein were mutated to produce a mutant, and the molecular docking showed the binding energies and the numbers of contact acid for 10 tetracyclines all increased. The mutant was coupled with Fe3O4 to synthesize a magnetic complex, and a fluorescent tracer was synthesized by coupling quantum dot and minocycline with bovine serum albumin. Under the assistance of 96-well bottom magnet, a semi-homogeneous method based on the two materials was developed on conventional microplate for determination of the 10 tetracyclines in milk. Results showed once assay was finished within 20 min, the limits of detection (drug concentration showing 10% inhibition) for the 10 drugs were in the range of 0.32-0.94 ng/mL, and the magnetic complex could be regenerated for 6 times. Furthermore, the sensitivities were improved for 4-6 folds in comparison with the use of natural TetR. Therefore, this method is simple, sensitive, time-saving and recyclable, and it can be used for routine screening of the 10 tetracyclines in milk.


Assuntos
Leite , Tetraciclinas , Animais , Tetraciclinas/análise , Leite/química , Simulação de Acoplamento Molecular , Antibacterianos/análise , Imunoensaio/métodos
3.
Front Microbiol ; 14: 1143017, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37152729

RESUMO

Plastoglobules, which are lipoprotein structures surrounded by a single hydrophobic phospholipid membrane, are subcellular organelles in plant chromoplasts and chloroplasts. They contain neutral lipids, tocopherols, quinones, chlorophyll metabolites, carotenoids and their derivatives. Proteomic studies indicated that plastoglobules are involved in carotenoid metabolism and storage. In this study, one of the plastid lipid-associated proteins (PAP), the major protein in plastoglobules, was selected and overexpressed in Phaeodactylum tricornutum. The diameter of the plastoglobules in mutants was decreased by a mean of 19.2% versus the wild-type, while the fucoxanthin level was increased by a mean of 51.2%. All mutants exhibited morphological differences from the wild-type, including a prominent increase in the transverse diameter. Moreover, the unsaturated fatty acid levels were increased in different mutants, including an 18.9-59.3% increase in eicosapentaenoic acid content. Transcriptomic analysis revealed that PAP expression and the morphological changes altered xanthophyll synthesis and storage, which affected the assembly of the fucoxanthin chlorophyll a/c-binding protein and expression of antenna proteins as well as reduced the non-photochemical quenching activity of diatom cells. Therefore, metabolic regulation at the suborganelle level can be achieved by modulating PAP expression. These findings provide a subcellular structural site and target for synthetic biology to modify pigment and lipid metabolism in microalgae chassis cells.

4.
Crit Rev Food Sci Nutr ; : 1-20, 2023 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-37114919

RESUMO

Nervonic acid, a 24-carbon fatty acid with only one double bond at the 9th carbon (C24:1n-9), is abundant in the human brain, liver, and kidney. It not only functions in free form but also serves as a critical component of sphingolipids which participate in many biological processes such as cell membrane formation, apoptosis, and neurotransmission. Recent studies show that nervonic acid supplementation is not only beneficial to human health but also can improve the many medical conditions such as neurological diseases, cancers, diabetes, obesity, and their complications. Nervonic acid and its sphingomyelins serve as a special material for myelination in infants and remyelination patients with multiple sclerosis. Besides, the administration of nervonic acid is reported to reduce motor disorder in mice with Parkinson's disease and limit weight gain. Perturbations of nervonic acid and its sphingolipids might lead to the pathogenesis of many diseases and understanding these mechanisms is critical for investigating potential therapeutic approaches for such diseases. However, available studies about this aspect are limited. In this review, relevant findings about functional mechanisms of nervonic acid have been comprehensively and systematically described, focusing on four interconnected functions: cellular structure, signaling, anti-inflammation, lipid mobilization, and their related diseases.

5.
Sci Total Environ ; 875: 162661, 2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-36898549

RESUMO

The paper discusses the implementation of Hong Kong's tailor-made sewage surveillance programme led by the Government, which has demonstrated how an efficient and well-organized sewage surveillance system can complement conventional epidemiological surveillance to facilitate the planning of intervention strategies and actions for combating COVID-19 pandemic in real-time. This included the setting up of a comprehensive sewerage network-based SARS-CoV-2 virus surveillance programme with 154 stationary sites covering 6 million people (or 80 % of the total population), and employing an intensive monitoring programme to take samples from each stationary site every 2 days. From 1 January to 22 May 2022, the daily confirmed case count started with 17 cases per day on 1 January to a maximum of 76,991 cases on 3 March and dropped to 237 cases on 22 May. During this period, a total of 270 "Restriction-Testing Declaration" (RTD) operations at high-risk residential areas were conducted based on the sewage virus testing results, where over 26,500 confirmed cases were detected with a majority being asymptomatic. In addition, Compulsory Testing Notices (CTN) were issued to residents, and the distribution of Rapid Antigen Test kits was adopted as alternatives to RTD operations in areas of moderate risk. These measures formulated a tiered and cost-effective approach to combat the disease in the local setting. Some ongoing and future enhancement efforts to improve efficacy are discussed from the perspective of wastewater-based epidemiology. Forecast models on case counts based on sewage virus testing results were also developed with R2 of 0.9669-0.9775, which estimated that up to 22 May 2022, around 2,000,000 people (~67 % higher than the total number of 1,200,000 reported to the health authority, due to various constraints or limitations) had potentially contracted the disease, which is believed to be reflecting the real situation occurring in a highly urbanized metropolis like Hong Kong.


Assuntos
COVID-19 , Humanos , COVID-19/epidemiologia , SARS-CoV-2 , Vigilância Epidemiológica Baseada em Águas Residuárias , Esgotos , Pandemias , Hong Kong/epidemiologia
6.
Huan Jing Ke Xue ; 44(2): 1095-1103, 2023 Feb 08.
Artigo em Chinês | MEDLINE | ID: mdl-36775632

RESUMO

The aim of this study was to understand the effect of deep vertical rotary tillage on the diversity and structural distribution of a soil bacterial community in cultivated land, as well as the interactions between microbial ecological molecular network and species. In this study, the cultivated soil under different tillage methods in the Yellow River diversion irrigation area in Ningxia was selected as the research object, and two treatments were set up, namely, deep vertical rotary tillage (DVRT) and conventional tillage (CT). Then, Illumina MiSeq technology was used to analyze the molecular ecological network of soil bacterial community under different tillage methods in the Ningxia Yellow River irrigation area. The results showed that DVRT significantly increased the contents of total organic carbon (TOC), total nitrogen (TN), alkali-hydrolyzed nitrogen (AN), total phosphorus (TP), available phosphorus (AP), and total potassium (TK) in the 0-20 cm soil layer compared with those in the CT treatment. Compared with those in the CT treatment, DVRT significantly increased the contents of TOC, TN, TP, AP, TK, and available potassium (AK) in the 20-40 cm soil layer. DVRT treatment significantly increased water content (WC) by 8.13%-13.30% and decreased pH and electrical conductivity (EC) by 4.51%-5.85% and 12.5%-13.33%, respectively. In different growth stages and soil layers of maize, the Shannon and Ace indices were increased in the DVRT treatments. The dominant bacterial phyla were:Actinobacteria, Proteobacteria, Chloroflexi, Acidobacteriota, and Gemmatimonadota. The results of bacterial community ß diversity showed that DVRT and CT had significant differences in bacterial community structure. The redundancy analysis and Partial Mantel test results showed that pH, EC, and TP were the key environmental factors affecting the structural diversity of bacterial communities. Molecular ecological network analysis revealed that DVRT treatment microbial networks consisted of more functionally related microbial modules, and the topological roles of key microorganisms were different from those in the CT treatment. In conclusion, DVRT can enhance soil nutrients and WC in different soil layers, reduce pH and EC, improve soil bacterial community diversity and microbial network structure, and also enhance the potential ecosystem functions in cultivated soil.


Assuntos
Ecossistema , Solo , Solo/química , Microbiologia do Solo , Bactérias/genética , Nitrogênio/análise , Fósforo , Potássio
7.
Lab Med ; 54(4): 411-423, 2023 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-36355716

RESUMO

OBJECTIVE: To establish a novel approach for diagnosing early- and midstage esophageal squamous cell carcinoma (ESCC). METHODS: The tumor suppressor gene phospholysine phosphohistidine inorganic pyrophosphate phosphatase (LHPP)-based miRNA signature was identified using next-generation sequencing and 3 biological online prediction systems. This retrospective study established and validated an ESCC prediction model using a test cohort and a validation cohort. RESULTS: Immunohistochemical staining and real-time quantitative polymerase chain reaction (RT-qPCR) results showed that LHPP protein levels were significantly lower in tissues with early- and midstage ESCC than in adjacent tissues (P < .01). Further, we confirmed that miR-15b-5p, miR-424-5p, miR-497-5p, miR-363-5p, and miR-195-5p inhibited LHPP. These 5 miRNAs were significantly elevated in the plasma of early- and midstage ESCC (P < .05). An ESCC prediction model combining these 5 miRNAs was established. Finally, in the external validation cohort, the model exhibited high discriminative value (sensitivity/specificity: 84.4%/93.3%). CONCLUSIONS: The prediction model has potential implications for diagnosis of early- and midstage ESCC.


Assuntos
Neoplasias Esofágicas , Carcinoma de Células Escamosas do Esôfago , MicroRNAs , Humanos , Estudos de Casos e Controles , Linhagem Celular Tumoral , População do Leste Asiático , Neoplasias Esofágicas/diagnóstico , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/diagnóstico , Carcinoma de Células Escamosas do Esôfago/genética , Carcinoma de Células Escamosas do Esôfago/patologia , Regulação Neoplásica da Expressão Gênica , Genes Supressores de Tumor , MicroRNAs/genética , Estudos Retrospectivos
8.
Int J Mol Sci ; 25(1)2023 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-38203518

RESUMO

Accumulating evidence shows that the abnormal increase in the mortality of intestinal epithelial cells (IECs) caused by apoptosis, pyroptosis, and necroptosis is closely related to the function of mucous membrane immunity and barrier function in patients with ulcerative colitis (UC). As a procedural death path that integrates the above-mentioned many deaths, the role of PANoptosis in UC has not been clarified. This study aims to explore the characterization of PANoptosis patterns and determine the potential biomarkers and therapeutic targets. We constructed a PANoptosis gene set and revealed significant activation of PANoptosis in UC patients based on multiple transcriptome profiles of intestinal mucosal biopsies from the GEO database. Comprehensive bioinformatics analysis revealed five key genes (ZBP1, AIM2, CASP1/8, IRF1) of PANoptosome with good diagnostic value and were highly correlated with an increase in pro-inflammatory immune cells and factors. In addition, we established a reliable ceRNA regulatory network of PANoptosis and predicted three potential small-molecule drugs sharing calcium channel blockers that were identified, among which flunarizine exhibited the highest correlation with a high binding affinity to the targets. Finally, we used the DSS-induced colitis model to validate our findings. This study identifies key genes of PANoptosis associated with UC development and hypothesizes that IRF1 as a TF promotes PANoptosome multicomponent expression, activates PANoptosis, and then induces IECs excessive death.


Assuntos
Colite Ulcerativa , Colite , Humanos , Colite Ulcerativa/genética , Apoptose , Biópsia , Bloqueadores dos Canais de Cálcio
9.
Neoplasma ; 69(1): 49-58, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34881626

RESUMO

The small nucleolar RNA host gene 16 (SNHG16) has recently been shown to be a putative oncogene in gastric cancer (GC) and other cancer types, but how its four lncRNA variants are expressed in any physiological and pathological situation remains unknown. To investigate the expression and function of the four lncRNA variants of SNHG16, mainly the variant 1, in GC, we performed quantitative PCR to determine the RNA levels of the four variants in 60 GC tissue samples and several cell lines. We also studied how knocking down of SNHG16 with siRNA affected proliferation, apoptosis, cell cycle progression, as well as migration and invasion of GC cells. Our results showed that variants 1 and 4 were overexpressed in GC tissues compared with adjacent uninvolved tissues. Knockdown of the four variants, mainly the variant 1, enhanced apoptosis and inhibited cell cycle progression of a GC cell line by arresting the cells at the G1 phase. These cellular effects were associated not only with decreased protein levels of c-Myc, PCNA, cyclins D1, E1, A2 and B, as well as CDKs 2 and 6, but also with increased protein levels of the p21, p27 and p53. Knockdown of total SNHG16 lncRNAs also inhibited invasion and migration of the GC cells in vitro. These results collectively suggest that SNHG16 may be oncogenic in GC by regulating cell cycle progression and may serve as a GC biomarker.


Assuntos
MicroRNAs , RNA Longo não Codificante , Neoplasias Gástricas , Ciclo Celular/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Oncogenes/genética , RNA Longo não Codificante/genética , Neoplasias Gástricas/genética
10.
Sci Total Environ ; 768: 144024, 2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-33736347

RESUMO

The disappearance of submerged aquatic vegetation caused by the use of purse seine in productive fishery has aroused the attention of the national government. In order to restore aquatic vegetation, the government has removed the seine nets on the Huayanghe Lake's surface. Here, fourteen cruises were conducted in the Huayanghe Lakes from 2016 to 2019 to study the response of water quality and phytoplankton communities to rapidly recovering aquatic vegetation. The results showed that the restoration of aquatic vegetation increased the Secchi depth (from 65.36 to 105.52 cm) and dissolved oxygen (from 8.98 to 12.17 mg/L), while the concentration of total nitrogen (from 0.98 to 0.53 mg/L), and total phosphorus (from 0.04 to 0.02 mg/L) decreased, resulting in a 35.3% and 11.6% reduction in the total phytoplankton density and chlorophyll ɑ, respectively. In addition, the restoration of aquatic vegetation significantly increased Shannon-Wiener diversity, Margalef richness indices by 51.6% and 40.1%. We also observed that phytoplankton density exhibited significant changes based on nonmetric multidimensional scaling analysis (NMDS). In Lake Huangda, the coverage of aquatic vegetation was as high as 80%. We also observed that after the restoration of aquatic vegetation, the proportion of cyanobacteria decreased significantly by 21%. Our study emphasizes that aquatic plants can alleviate eutrophication and increase the diversity of phytoplankton, thus providing guidance for the restoration and protection of water ecosystems in the lakes connected to the Yangtze River.


Assuntos
Lagos , Fitoplâncton , China , Ecossistema , Eutrofização , Fósforo/análise , Qualidade da Água
11.
Exp Cell Res ; 391(1): 111980, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32229193

RESUMO

BACKGROUND: Prodrugs are medications or compounds that, after administration, can be converted into pharmacologically active drugs through metabolism. Unlike conventional drugs, prodrugs have reduced adverse or unintended effects, which could become critical limitations in treatments such as chemotherapy. Previously through computer-aided drug design and chemical synthesis, we have obtained and examined a prodrug N-benzyloxycarbonyl-Ala-Asn-Doxorubicin (CBZ-AAN-DOX). CBZ-AAN-DOX is essentially Doxorubicin that is chemically-modified with tripeptides to target Legumain, a highly expressed protein in cancer cells and is involved in tumor metastasis and tumor microvessel formation. The difficulty to test the safety and efficacy of the prodrug (including the pharmacodynamic parameters of CBZ-AAN-DOX on metastasis and invasion of tumors, as well as cardiac and vascular toxicity) primarily comes from the lack of appropriate experimental models. METHODS: Human cervical cancer cell lines CaSki under hypoxic conditions were used to evaluate the cell viability by CCK-8 assay after the prodrug treatment. Western blotting method was performed for Legumain protein determination in the cell culture. Wound healing and transwell invasion assays were performed to determine the effects of the prodrug on tumor metastasis and invasion, respectively. Zebrafish models were constructed for toxicity and angiogenesis visual analysis after in vivo treatment with the prodrug. RESULTS: The CCK-8 results showed that CBZ-AAN-DOX exhibits an IC50 of 28.7 µM in 48 h on CaSki cells that had a lower cell inhibition rate than DOX 80.3 µM for 24 h. Legumain expression was significantly increased in a time-dependent manner in 48 h under hypoxia conditions. The results also showed that 13.9 µM of the prodrug significantly inhibited the migration and invasion of cells and the effects were significantly stronger than that of 41.8 µM of DOX under hypoxia conditions after 48 h. The effects of 160 µM of the prodrug on the survival rate of zebrafish after 72 h and heart-toxicity showed no obvious abnormalities. Cell metastasis and angiogenesis were also inhibited in tumor-bearing zebrafish model. CONCLUSION: The findings in this study demonstrated that CBZ-AAN-DOX is a promising chemotherapy candidate with low toxicity and high efficiency for cervical cancer. Remarkably, the hypoxic culture model together with the zebrafish model serve as a good system for the evaluation of the toxicity, targeting and impact of the prodrug on tumor invasion and metastasis.


Assuntos
Antibióticos Antineoplásicos/farmacologia , Cisteína Endopeptidases/genética , Doxorrubicina/análogos & derivados , Regulação Neoplásica da Expressão Gênica , Neovascularização Patológica/prevenção & controle , Oligopeptídeos/farmacologia , Pró-Fármacos/farmacologia , Neoplasias do Colo do Útero/tratamento farmacológico , Animais , Antibióticos Antineoplásicos/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células , Cisteína Endopeptidases/metabolismo , Modelos Animais de Doenças , Doxorrubicina/metabolismo , Doxorrubicina/farmacologia , Feminino , Humanos , Hipóxia/diagnóstico , Hipóxia/tratamento farmacológico , Hipóxia/genética , Hipóxia/mortalidade , Neovascularização Patológica/diagnóstico , Neovascularização Patológica/genética , Neovascularização Patológica/mortalidade , Oligopeptídeos/metabolismo , Pró-Fármacos/metabolismo , Análise de Sobrevida , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/mortalidade , Ensaios Antitumorais Modelo de Xenoenxerto , Peixe-Zebra
12.
Explore (NY) ; 16(2): 110-115, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31492552

RESUMO

OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of motherwort injection (MI, Yi mu cao) in preventing post-abortion hemorrhage and promoting uterine retraction after induced abortion surgery. METHODS: In this multi-center randomized controlled trial undertaken between September 2014 and August 2016, 408 child-bearing age women who underwent induced abortion surgery and randomly assigned to two groups: MI group who received intramuscular MI treatment (n = 207) and no-treatment group (n = 201). The volume of vaginal bleeding and its duration were used as primary end points; and uterine size and dimension were measured as secondary end points. Blood coagulation indices, routine blood and adverse events were recorded to evaluate the safety. RESULTS: The efficacy analysis was based on 366 patients. No significant difference were found in baseline characteristics between the two groups (P > 0.05). The amount of vaginal bleeding and duration of bleeding were significantly reduced in MI group compared with on-treatment group in Full Analysis Set (FAS) and Per Protocol Set (PPS) populations (P < 0.001). The volume and perimeter of the uterine dimensions in MI group were smaller than no-treatment group (P < 0.001). No significant abnormal vital signs were observed. Only 1 case of mild, transient erythema was found in MI group. CONCLUSIONS: MI could reduce post-abortion hemorrhage and improve uterine retraction in women undergoing surgical induced abortion, without serious adverse events. It was a safe and effective treatment in preventing post-abortion hemorrhage after induced abortion.


Assuntos
Aborto Induzido/métodos , Leonurus/química , Ocitócicos/administração & dosagem , Hemorragia Uterina/prevenção & controle , Adulto , China , Feminino , Humanos , Injeções Intramusculares , Gravidez , Útero/efeitos dos fármacos
13.
Zool Res ; 41(1): 78-83, 2020 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-31746565

RESUMO

Individual dispersal trends, unquestionably important for species ecology and evolution, are affected by multiple factors. Understanding the factors that influence female dispersal strategies offers important insight into primate dispersal mechanisms and female choice. To investigate the proximate causes of dispersal in female Yunnan snub-nosed monkeys (Rhinopithecus bieti), we observed and analyzed nine years of detailed dispersal and demographic data from a population of R. bieti in Xiangguqing, Baimaxueshan Nature Reserve, Yunnan Province, China. Results showed that females who lived long-term in a one-male unit (OMU), without giving birth and with few or no relatives, were more likely to leave that OMU. In addition, an OMU led by an outgroup male and containing more female relatives was significantly more likely to be chosen for immigration. Conversely, greater male age, longer male tenure, and more potentially fertile females discouraged immigration into an OMU. These results suggest that reproduction, male quality, and kin cooperation play the largest roles in female Yunnan snub-nosed monkey dispersal.


Assuntos
Distribuição Animal , Comportamento Animal , Presbytini/fisiologia , Comportamento Social , Envelhecimento , Animais , Feminino , Masculino
14.
Int J Mol Med ; 44(2): 417-426, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173172

RESUMO

Myocardial injury is one of the main symptoms of sepsis. However, the mechanisms underlying sepsis­induced myocardial dysfunction remain unclear. In the present study, the concentration of cardiac troponin T (CTnT) in serum was measured using an enzyme­linked immunosorbent assay kit. The levels of interleukin (IL)­1ß and IL­18 were assessed by reverse transcription­quantitative polymerase chain reaction (RT­qPCR) analysis and the level of malondialdehyde (MDA) was determined using a corresponding kit. Myocardial pathology was analyzed via hematoxylin and eosin staining. RT­qPCR analysis and western blotting and/or immunohistochemistry were used to quantify the expression levels of thioredoxin­interacting protein (TNXIP), NOD­like receptor pyrin domain containing 3 (NLRP3), cleaved caspase­1, caspase­1, catalase and manganese­superoxide dismutase (MnSOD). The viability of cells was determined using a cell counting kit­8. Apoptosis and reactive oxygen species (ROS) were examined using flow cytometry. Models of sepsis­induced myocardial injury were successfully established; evidence included increases in the levels of CTnT, IL­1ß, IL­18 and MDA and myocardial tissue damage in vivo, and decreased cell viability and improvements in IL­1ß and IL­18 in vitro. The levels of TXNIP, NLRP3 and cleaved caspase­1 were upregulated in the sepsis models. Small interfering RNA targeting TNXNIP (siTXNIP) increased cell viability, reduced the apoptotic rate and attenuated the release of IL­1ß and IL­18. The levels of TXNIP, NLRP3 and cleaved caspase­1 and production of ROS were suppressed by siTXNIP, accompanied by increases in catalase and MnSOD. TXNIP/NLRP3 serves an important role in the development of sepsis­induced myocardial damage.


Assuntos
Proteínas de Ciclo Celular/análise , Cardiopatias/etiologia , Miocárdio/patologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/análise , Sepse/complicações , Animais , Apoptose , Linhagem Celular , Cardiopatias/patologia , Masculino , Ratos Sprague-Dawley , Sepse/patologia
15.
Biochem Biophys Res Commun ; 513(4): 891-897, 2019 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-31003768

RESUMO

BACKGROUND: Disulfiram (DSF), a drug widely used to control alcoholism, which has anticancer activity by inducing apoptosis in a copper (Cu)-dependent manner. Numerous evidences from mouse experiments indicated that some anti-cancer agents of chemotherapeutic drugs favor the induction of immunogenic cancer cell death (ICD) leading to tumor-specific immune responses. However, whether DSF could induce the colorectal tumor cells death and the mechanism involved in ICD regulatory remains elusive. The main objective of this study was to elucidate the effect of DSF/Cu on the apoptosis of colorectal cancer (CRC) cells and the expression of the two major ICD markers in CRC cells: calreticulin (CRT) and heat shock proteins (HSP) 70. METHODS: Firstly, the toxicity of DSF/Cu in HCT116, SW620 and HCT8 cells was assayed by MTT. Flow cytometry was utilized to detect the apoptosis effects. The effects of DSF/Cu on the expression of ICD-related molecules in tumor tissues were further verified in the CRC xenograft mouse model. RESULTS: The results showed that DSF/Cu increase apoptosis of these three cells in a dose dependent manner and significantly inhibited the proliferation at the concentration range from 0.05 to 1.6 µM. Furthermore, the expression of CRT and HSP70 on the cell surface also increased. The rate of transplanted tumors grew slowly, and the expression of CRT and HSP70 in colorectal cancer tissues was increased after treated with DSF/Cu. CONCLUSION: In conclusion, our results show that DSF/Cu exerts anti-colorectal cancer and its underlying mechanisms are associated with the enhancement of molecules expression of cell ICD. These results provide experimental evidence and theory basis of therapy for developing the DSF/Cu as the drug for CRC.


Assuntos
Neoplasias Colorretais/tratamento farmacológico , Dissulfiram/farmacologia , Morte Celular Imunogênica/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Calreticulina/metabolismo , Linhagem Celular Tumoral , Neoplasias Colorretais/patologia , Cobre/farmacologia , Dissulfiram/uso terapêutico , Dissulfiram/toxicidade , Células HCT116 , Proteínas de Choque Térmico HSP70/metabolismo , Xenoenxertos , Humanos , Camundongos
16.
Open Life Sci ; 14: 217-223, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33817155

RESUMO

OBJECTIVE: This study aims to investigate the differences in the expression of hypoxia-inducible factor-1α (HIF-1α), N-myc downstream-regulated gene 2 (NDRG2) and epithelial mesenchymal transition (EMT)-related proteins in normal gastric tissues, gastric cancer tissues and lymph node metastasis. METHODS: Immunohistochemistry was used to detect the expression of HIF-1α, NDRG2, E-cadherin, Snail and Twist in normal gastric tissues, gastric cancer tissues and lymph node metastasis. RESULTS: In normal gastric tissues, HIF-1α was not expressed, NDRG2 was highly expressed. There was a significant between the expression of NDRG2 and Snail, as well as of NDRG2 and Twist. In gastric cancer tissues, there was no statistically difference between the expression of HIF-1α and E-cadherin, NDRG2 and E-cadherin. However, there was a significant difference in expression between the expression of HIF-1α and Snail, HIF-1α and Twist, NDRG2 and Snail, and NDRG2 and Twist. In lymph node metastasis tissues, we show that HIF-1α was highly expressed, while NDRG2 was not, and the difference between the expression of HIF-1α and E-cadherin, HIF-1α and Snail, HIF-1α and Twist was not significant. CONCLUSION: HIF-1α may promote EMT, possibly by inhibiting the expression of NDRG2.

17.
Luminescence ; 34(1): 98-105, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30548788

RESUMO

The residues of phenothiazines and benzodiazepines in foods of animal origin are dangerous to consumers. For inspection of their abuses, this study for the first time reported on the use of a chemiluminescence array sensor for the simultaneous determination of four phenothiazines and five benzodiazepines in pig urine. Two molecularly imprinted polymers were coated in different wells of a conventional 96-well microtiter plate as the recognition reagents. After sample loading, the absorbed analytes were initiated directly by using an imidazole enhanced bis(2,4,6-trichlorophenyl)oxalate-hydrogen peroxide system to emit light. The assay process consisted of only one sample-loading step prior to data acquisition, so one test was finished within 10 min. The limits of detection for the nine drugs in the pig urine were in a range of 0.1 to 0.6 pg/mL, and the recoveries from the fortified blank urine samples were in a range of 80.3 to 95%. Furthermore, the sensor could be reused six times. Therefore, this sensor could be used as a simple, rapid, sensitive and reusable tool for routine screening for residues of phenothiazines and benzodiazepines in pig urine.


Assuntos
Benzodiazepinas/urina , Medições Luminescentes/métodos , Fenotiazinas/urina , Polímeros/química , Animais , Desenho de Equipamento , Peróxido de Hidrogênio/química , Limite de Detecção , Medições Luminescentes/instrumentação , Microscopia Eletrônica de Varredura , Impressão Molecular , Nitrazepam/química , Oxalatos/química , Prometazina/química , Sensibilidade e Especificidade , Suínos , Fatores de Tempo
18.
Front Pharmacol ; 9: 1292, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30459625

RESUMO

Cannabinoid receptor 2 (CB2) has been reported to produce a cardio-protective effect in cardiovascular diseases such as myocardial infarction. Here in this study, we investigated the role of CB2 in diabetic cardiomyopathy (DCM) and its underlying mechanisms. HU308 was used for the selective activation of CB2. Bafilomycin A1 was used for the blockade of autophagy and compound C was used to inhibit AMPK signaling. An streptozotocin (STZ)-induced mice model and high glucose (HG)-challenged cardiomyocytes were applied for study. Cardiac function was detected by echocardiography and Western blot for the detection of autophagy-related and its signaling-related proteins. Transmission electron microscopy was used for the analysis of autophagosome number. Cell viability was detected by Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) release assays. We found that activating CB2 by HU308 improved cardiac function in DCM as well as cell viability in cardiomyocytes under HG challenge, while the administration of bafilomycin A1 attenuated the protective effects. HU308 enhanced the level of autophagy in the heart tissues from DCM mice as well as cardiomyocytes under HG challenge. HU308 triggered the AMPK-mTOR-p70S6K signaling pathway, while the administration of compound C attenuated the cardio-protective effect of HU308 in cardiomyocytes under HG challenge. In conclusion, we initially demonstrated that activating CB2 produced a cardio-protective effect in DCM as well as cardiomyocytes under HG challenge through inducing the AMPK-mTOR-p70S6K signaling-mediated autophagy.

19.
Anal Biochem ; 554: 9-15, 2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-29803786

RESUMO

In this study, a molecularly imprinted polymer based chemiluminescence array capable of simultaneous determining phenothiazines and benzodiazepines was first reported. Two polymers were coated in different wells of the conventional 96-well microtiter plate as the recognition reagents, and the added analytes competed with a horseradish peroxidase-labeled bi-hapten conjugate to bind the recognition reagents. The light signal was induced by using a highly effective luminol-H2O2-IMP system. The assay procedure consisted of only one sample-loading step prior to data acquisition. Then, the array was used to determine 4 phenothiazines and 5 benzodiazepines in pork simultaneously. The limits of detection for the 9 drugs were in a range of 0.001-0.01 ng/mL, and the recoveries from the fortified blank pork were in a range of 63.5%-94.1%. Furthermore, the array could be reused for 8 times. The detection results for some real pork samples were consistent with an ultra performance liquid chromatography method.


Assuntos
Benzodiazepinas/análise , Resíduos de Drogas/análise , Contaminação de Alimentos/análise , Fenotiazinas/análise , Carne Vermelha/análise , Animais , Cromatografia Líquida , Ensaio de Imunoadsorção Enzimática , Humanos , Limite de Detecção , Medições Luminescentes/métodos , Impressão Molecular/métodos , Polímeros/química , Sus scrofa
20.
PLoS One ; 12(5): e0174612, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28472139

RESUMO

Lean-type Pekin duck is a breed gained through long-term selection and great effort has been exerted to understand the mechanisms underlying increased muscle yields. However, the genes involved in Pekin duck embryonic breast muscle development have not been explored to date. In this study, we investigated gene expression profiles in Pekin Duck embryonic breast muscle at hatched day 13 (E13), E19, and E27 using RNA-seq. In total, we produced 519,312,178 raw reads resulting in 497,348,158 high-quality reads after filtering. The mapping, distribution of reads along annotated genes, and consistency across replicates demonstrates the high quality of the RNA-seq data used in this study, allowing us to continue with the downstream analysis. Significantly fewer differentially expressed genes (DEGs) were identified between E13 and E19 (203 DEGs) compared to E27 and E19 (2,797 DEGs). Many DEGs highly expressed in E19 are involved in metabolic processes and cell division. KEGG analysis showed many pathways associated with fat development were significantly enriched for DEGs highly expressed in E27. These results provide a basis for the further investigation of the mechanisms involved in Pekin duck embryonic breast muscle development.


Assuntos
Patos/embriologia , Perfilação da Expressão Gênica , Músculo Esquelético/embriologia , Animais , Análise de Sequência de RNA
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