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In vivo models of brain pathology are crucial for studying neurological diseases. Here, we present a protocol to induce a pathological condition in a mouse brain area by local injection of neurotoxic stimulus. We describe steps for preparing reagents, stereotaxic injection procedures to induce neurodegeneration in the hippocampus, and preparation of brain sections to examine the induced model. This protocol is useful for studying how local pathology affects other brain areas and neighbor cells and its functional consequences in behavior. For complete details on the use and execution of this protocol, please refer to Zhang et al.1.
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Background: Nonalcoholic fatty liver disease (NAFLD) has become a major global health burden, which increases the risk of extra-hepatic complications such as type 2 diabetes mellitus (T2DM), dyslipidemia, metabolic syndrome (MetS), and cardiovascular disease (CVD). However, NAFLD remains underappreciated and underdiagnosed. Our study aimed to explore the prevalence of NAFLD and the association between NAFLD and CVD events among adults aged 40 and older in Northern China. Methods: This study was conducted in the Shijingshan district of Beijing, China from November 2011 to August 2012. A total of 18891 subjects were recruited in the study. The information including demographical information, lifestyle, previous history of diabetes, hypertension, dyslipidemia, CVD, and liver disease were gathered. Data on physical examination, blood lipid profile, fasting blood glucose, and 2-hour blood glucose were recorded. Determination of MetS was according to T2DM guideline of Chinese Diabetes Society (2020 edition). The association between CVD and NAFLD was evaluated by multivariate logistic regression. Results: The prevalence of NAFLD was 15.2%. After adjustment for age, smoking status, alcohol intake, WC, hypertension, dyslipidemia and hyperglycemia, the odds ratios (ORs) of CVD in men were 1.622 (95%CI: 1.345-1.957) and 1.990 (95%CI: 1.709-2.316) in women with NAFLD, compared with the subjects without NAFLD. Conclusions: NAFLD is independently associated with increased risk of CVD development.
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PURPOSE: To clarify the effect of genistein(GEN) on osteogenic differentiation and explore the effect of GEN loaded by platelet-rich fibrin (PRF) on the repair process of bone defects in obese mice. METHODS: In in vitro experiments, the effect of GEN(0, 0.1, 1, 10, 50 µmol/L) on the proliferation of mouse embryonic osteoblast precursor cells (MC3T3-E1) was determined by CCK 8. Alkaline phosphatase(ALP) staining and quantitative detection of ALP activity were performed to determine the changes of ALP activity in cells; RNA and protein expression levels of ALP, osteopontin (OPN) and osteocalcin (OCN) were detected by quantitative real-time PCR(qRT-PCR) and Western blot. Alizarin red staining was used to define the effect of GEN on mineralization of MC3T3-E1. To verify the feasibility of the PRF drug loading, the ultrastructure of PRF was subsequently observed under SEM. In in vivo experiments, obese C57 mouse models were established by high-fat diet feeding. On this basis, skull defect models with a diameter of 2.8 mm were established, and the prepared GEN/PRF complexes were placed into the bone defect area. The effects of GEN on skull defect repair in obese mice were evaluated by Micro-CT scanning and hematoxylin-eosin(H-E) staining. Statistical analysis was performed with GraphPad Prism 5.0 software package. RESULTS: CCK 8 results showed that 0.1, 1 µmol/L GEN promoted cell proliferation within 7 days(Pï¼0.05); 10 µmol/L GEN had no significant effect on the process of cell proliferation. From the second day, 50 µmol/L GEN significantly inhibited cell growth and showed cytotoxicity(Pï¼0.05). These two concentrations had similar effects in promoting cellular osteogenic differentiation. SEM results showed that PRF presented a 3-dimensional network structure, providing space for loading drug molecules. In in vivo experiments, the body weight of mice in the high-fat diet (HFD) group was 27.7% greater than that in the normal diet group(Pï¼0.05) and had abnormal glucose tolerance (Pï¼0.05). Micro-CT showed that compared with the normal diet group, the number of bone trabeculae in the femur of obese mice was decreased(Pï¼0.05), the distance between bone trabeculae was widened(Pï¼0.05), and the bone density was decreased (Pï¼0.05). In addition, GEN (0.1, 1.0 µmol/L) loaded by PRF increased bone volume fraction in the skull of obese mice (Pï¼0.05). H-E results showed that GEN/PRF promoted the healing of the bone defects. CONCLUSIONS: GEN promotes osteogenic differentiation of MC3T3-E1, and it can effectively accelerate the healing of cranial bone defects after loading with PRF in obese mice.
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Osteogênese , Fibrina Rica em Plaquetas , Animais , Camundongos , Osteogênese/genética , Genisteína/farmacologia , Camundongos Obesos , Sincalida/farmacologia , Diferenciação Celular/genética , OsteoblastosRESUMO
Circadian clocks, generating daily rhythms in biological processes, maintain homeostasis in physiology, so clock alterations are considered detrimental. Studies in brain pathology support this by reporting abnormal circadian phenotypes in patients, but restoring the abnormalities by light therapy shows no dramatic effects. Recent studies on glial clocks report the complex effects of altered clocks by showing their beneficial effects on brain repairs. However, how neuronal clocks respond to brain pathology is elusive. This study shows that neuronal BMAL1, a core of circadian clocks, reduces its expression levels in neurodegenerative excitotoxicity. In the dentate gyrus of excitotoxic hippocampal lesions, reduced BMAL1 in granule cells precedes apoptosis. This subsequently reduces BMAL1 levels in neighbor neural stem cells and progenitors in the subgranular zone, enhancing proliferation. This shows the various BMAL1 roles depending on cell types, and its alterations can benefit brain repair. Thus, cell-type-specific BMAL1 targeting is necessary to treat brain pathology.
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Background: Quantification of vertebral arteries can provide insights into basilar curvature and plaque. Therefore, this retrospective study aimed at identifying the dominant vertebral artery (VA) causing basilar artery (BA) curvature and to further quantify the effect of dominant VA angle on BA curvature and BA plaque using high-resolution magnetic resonance imaging (HRMRI) and 3-dimensional time-of-flight magnetic resonance angiography (3D-TOF-MRA). Methods: This retrospective analysis included 521 participants who underwent HRMRI in the China-Japan Friendship Hospital from November 2015 to October 2021 for neurological symptoms or signs. The VA angle more related to BA curvature was defined as the dominant VA angle. Multivariable linear regression analysis was used to evaluate the relationship between the dominant VA angle and mid-BA angle, while multivariable logistics regression was used to evaluate the influence of the dominant VA angle and clinical risk factors on BA plaque. Results: In total, 259 participants were included in this study (mean age 53.71±13.12 years; 146 males). The balanced-type participants had a significantly lower probability of BA plaques (P<0.001). The Chi-squared test showed that the BA curvature direction was significantly associated with the side with larger VA diameter (P<0.001) and larger VA angle (P<0.001). As a result, the VA angle on the side with the larger diameter or the larger VA angle when the diameters were similar was considered to be the dominant VA angle. The dominant VA angle was independently correlated with the mid-BA angle (P<0.001). In addition, the dominant VA angle was also an independent risk factor for BA plaque. Additionally, 80° was the cutoff value of the dominant VA angle, and when the dominant VA angle was greater than 80°, the risk of BA plaque increased about 18-fold [odds ratio, 18.951; 95% confidence interval (CI): 4.545-79.026; P<0.001]. Conclusions: The dominant VA angle was independently associated with BA plaque, and a dominant VA angle greater than 80° may be a marker for a high risk of posterior circulation atherosclerosis.
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Postoperative pulmonary complications (PPCs) are among the most common complications after cardiovascular surgery. This study aimed to explore the real incidence of and risk factors for PPC in patients with acute type A aortic dissection (ATAAD) who underwent total aortic arch replacement combined with the frozen elephant trunk (TAR + FET). In total, 305 ATAAD patients undergoing TAR + FET from January 2021 to August 2022 in a single-center were divided into PPCs or non-PPCs group. The incidence of PPCs was calculated, risk factors of PPCs were analyzed, and postoperative outcomes were compared between these 2 groups. The incidence of any PPC was 29.2%. And the incidence of respiratory infection, respiratory failure, pleural effusion, atelectasis, pneumothorax, acute respiratory distress syndrome, aspiration pneumonitis, pulmonary edema and bronchospasm was 23.0%, 12.5%, 10.5%, 1.0%, 0.7%, 1.0%, 0%, 0.7%, 0%, respectively. The logistic regression analysis revealed that the history of diabetes, history of renal dysfunction, preoperative SpO2 <90%, cardiopulmonary bypass duration, fresh frozen plasma volume and platelet concentrates volume were independent risk factors for PPCs. Among 2 groups, postoperative ventilation duration, postoperative length of stay in intensive care unit and hospital were (73.5 ± 79.0 vs 24.8 ± 35.2 hours; P < .001), (228.3 ± 151.2 vs 95.2 ± 72.0 hours; P < .001) and (17.9 ± 8.8 vs 11.5 ± 6.2 days; P < .001). There was no difference between 2 groups of in-hospital mortality rate. Additionally, other short-term outcomes were also significantly poorer in patients with PPCs. PPCs are common in ATAAD patients undergoing TAR + FET, and could be multifactorial. PPCs occurrence are associated with poor patient outcomes postoperatively and worth further investigation.
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Pneumotórax , Atelectasia Pulmonar , Humanos , Estudos Retrospectivos , Pulmão , Fatores de Risco , Pneumotórax/etiologia , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologiaRESUMO
A simple dual-signal assay that combined colorimetric and fluorometric strategy for uric acid (UA) rapid detection was designed based on the versatility of facile synthesized MnO2 nanosheet. The oxidization of 3,3',5,5'-tetramethylbenzidine (TMB) and the fluorescence quenching of quantum dots (QDs) occurred simultaneously in the presence of MnO2 nanosheet. UA could decompose MnO2 nanosheet into Mn2+, resulting in the fluorescence recovery of QDs, along with the fading of the blue color of ox TMB. Based on the principles above, the detection of UA could be realized by the change of the dual signals (colorimetric and fluorometric). The linear range of the colorimetric mode was 5-60 µmol L-1, and the limit of detection (LOD) was 2.65 µmol L-1; the linear range of the fluorescence mode was wide at 5-120 µmol L-1, and the LOD could be as low as 1.33 µmol L-1. The method was successfully used for analyzing UA levels in human serum samples, indicating that this new dual-signal method could be applied in clinical diagnosis.
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Nanoestruturas , Óxidos , Humanos , Ácido Úrico , Colorimetria/métodos , Compostos de ManganêsRESUMO
The growing demand for wireless connectivity is attracting interest in optical wireless communication (OWC) technique. In this paper, a filter-aided crosstalk mitigation scheme, employing digital Nyquist filters, is proposed to eliminate the trade-off between the spatial resolution and the channel capacity for the AWGR-based 2D infrared beam-steered indoor OWC system. By shaping the transmitted signal for narrow spectral occupancy, the inter-channel crosstalk resulting from the imperfect AWGR filtering can be avoided, which enables a denser AWGR grid. In addition, the spectral-efficient signal reduces the bandwidth requirement of the AWGR, which allows a low-complexity AWGR design. Thirdly, the proposed method is not sensitive to the wavelength misalignment between AWGRs and lasers, which relaxes the design of high wavelength stability lasers. Moreover, the proposed method is cost-efficient as we can make use of the mature DSP technique without additional optical components. The 20-Gbit/s data rate OWC capacity using PAM4 format has been experimentally demonstrated over a 6-GHz bandwidth-limited AWGR-based 1.1-m free-space link. The experimental results show the feasibility and effectiveness of the proposed method. By combining our proposed method with the polarization orthogonality technique, a promising capacity per beam of 40 Gbit/s is potentially attainable.
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AIMS: The basilar artery (BA) geometry and plaque characteristics may play an important role in the development of atherosclerosis. This study was performed to explore the relationship between the mid-BA angle and plaque characteristics and its effect on pontine infarction using high-resolution magnetic resonance imaging and three-dimensional time-of-flight magnetic resonance angiography. METHODS: In total, 77 patients with BA plaques were included in this study. According to the presence of acute pontine infarction on diffusion-weighted imaging, the patients were divided into a pontine infarction group and pontine non-infarction group. The mid-BA angle, plaque burden, stenosis ratio, positive remodeling, and intraplaque hemorrhage were evaluated to investigate their effects on stroke. RESULTS: The pontine infarction group had a greater plaque burden, stenosis ratio, positive remodeling, and mid-BA angle than the pontine non-infarction group. The correlation between the plaque burden and mid-BA angle was the highest (r=0.441, Pï¼0.001). Multivariate logistic regression analysis showed that the plaque burden (odds ratio, 1.164; 95% confidence interval, 1.093-1.241; Pï¼0.001) was an independent risk factor for pontine infarction. CONCLUSION: The mid-BA angle may increase the incidence of pontine infarction by increasing the plaque burden.
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Aterosclerose , Infartos do Tronco Encefálico , Placa Aterosclerótica , Humanos , Artéria Basilar/diagnóstico por imagem , Artéria Basilar/patologia , Constrição Patológica/complicações , Placa Aterosclerótica/complicações , Placa Aterosclerótica/diagnóstico por imagem , Placa Aterosclerótica/patologia , Aterosclerose/patologia , Imageamento por Ressonância Magnética/métodos , Infartos do Tronco Encefálico/complicações , Infartos do Tronco Encefálico/diagnóstico por imagemRESUMO
This paper mainly explores the complex impacts of spatial heterogeneity, vector-bias effect, multiple strains, temperature-dependent extrinsic incubation period (EIP) and seasonality on malaria transmission. We propose a multi-strain malaria transmission model with diffusion and periodic delays and define the reproduction numbers Ri and R^i (i = 1, 2). Quantitative analysis indicates that the disease-free ω-periodic solution is globally attractive when Ri<1, while if Ri>1>Rj (i≠j,i,j=1,2), then strain i persists and strain j dies out. More interestingly, when R1 and R2 are greater than 1, the competitive exclusion of the two strains also occurs. Additionally, in a heterogeneous environment, the coexistence conditions of the two strains are R^1>1 and R^2>1. Numerical simulations verify the analytical results and reveal that ignoring vector-bias effect or seasonality when studying malaria transmission will underestimate the risk of disease transmission.
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Malária , Modelos Biológicos , Humanos , Número Básico de Reprodução , Malária/epidemiologia , Difusão , TemperaturaRESUMO
Melanoma is the most aggressive form of skin cancer with the poorest prognosis and its pathogenesis has yet to be fully elucidated. As key factors that regulate cellular homeostasis, both reactive oxygen species (ROS) and autophagy are involved in the development of melanoma, from melanomagenesis to progression and drug resistance. However, the interaction between ROS and autophagy in the etiology and treatment of melanoma is not well characterized. The present review examined the production of ROS and the role of oxidative stress in melanoma, and summarized the role of ROSmediated autophagy in melanomagenesis and melanoma cell fate decision following treatment with various anticancer drugs. The present findings may lead to a better understanding of the pathogenesis and progression of melanoma, and suggest promising treatment options for this disease.
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Antineoplásicos , Melanoma , Antineoplásicos/farmacologia , Autofagia , Humanos , Melanoma/patologia , Estresse Oxidativo , Espécies Reativas de OxigênioRESUMO
Simultaneous analysis of mRNAs and proteins at the single-cell level provides information about the dynamics and correlations of gene and protein expressions in individual cells, enabling a comprehensive study of cellular heterogeneity and expression patterns. Here, we present a platform for about 1000 cellular indexing of mRNAs and membrane proteins, named multi-Paired-seq, with high cell utilization, accurate molecular measurement, and low cost. Based on hydrodynamic differential flow resistance, multi-Paired-seq largely improves cell utilization in the percentage of cells measured in population (>95%). Combined with the pump/valve structure, cell-free antibodies and mRNAs can be removed completely for highly accurate detection (R = 0.96) of protein copies. The picoliter reaction chambers allow high detection sensitivity for both mRNA transcripts and protein copies and low sequencing cost. Using multi-Paired-seq, three clusters of known breast cancer cell types are identified according to multimodal measurements, and the expression correlations between mRNAs and proteins under altered conditions are quantified. Multi-Paired-seq provides multimodal measurements at the single-cell level, which offers a new tool for cell biology, developmental biology, drug discovery, and precision medicine.
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Medicina de Precisão , Transcriptoma , Perfilação da Expressão Gênica , RNA Mensageiro/metabolismo , Análise de Sequência de RNA , Análise de Célula ÚnicaRESUMO
Background: Obesity can lead to the development of type 2 diabetes mellitus (T2DM). However, the predictive power of different obesity anthropometric indices (ObAIs) for T2DM varies with race and geographical area. Therefore, the study aimed to investigate the association between different ObAIs and T2DM and determine the best index for screening T2DM in middle-aged and elderly men and women in Beijing, China. Methods: A cross-sectional study was conducted in Shijingshan district (Beijing, China) from November 2011 to August 2012, involving a total of 14,558 subjects aged ≥40 years. Data on demographic information, lifestyle, history of T2DM, hypertension and dyslipidemia were collected. Body height, body weight, waist circumference (WC), hip circumference, and blood pressure were recorded. The oral glucose tolerance test or a standard meal test and blood lipid test were performed. The relationship between different ObAIs and T2DM was analyzed using multiple logistic regression. Results: After adjustment for age, smoking status, alcohol intake, occupation and education degree, diabetes family history, hypertension and dyslipidemia, body mass index (BMI), WC, waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR) were positively associated with T2DM in both men and women. Compared with the lowest BMI, WC, WHR, and WHtR quartiles, ORs of the highest quartiles were 2.131 (95% CI: 1.465-3.099), 1.752 (95% CI: 1.270-2.417), 1.342 (95% CI: 1.072-1.678), 2.739 (95% CI: 2.293-3.271) in men and 1.837 (95% CI: 1.584-2.130), 3.122 (95% CI: 1.980-4.924), 3.781 (95% CI: 2.855-5.007), 2.379 (85% CI: 2.040-2.775), respectively, in women. The areas under ROC curve of BMI, WC, WHR, and WHtR for men were 0.584 (95% CI: 0.568-0.600), 0.509 (95% CI: 0.492-0.525), 0.501 (95% CI: 0.485-0.518), and 0.642 (95% CI: 0.627-0.658) and 0.619 (95% CI: 0.607-0.632), 0.709 (95% CI: 0.697-0.720), 0.741 (95% CI: 0.730-0.752), and 0.654 (95% CI: 0.642-0.666), respectively, for women. Conclusion: WHtR and WHR have been found to perform better as predictors of T2DM in middle-aged and elderly Chinese men and women, respectively.
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BACKGROUND: Leaf senescence, the final stage of leaf growth and development, is regulated by numerous internal factors and environmental cues. Ethylene is one of the key senescence related hormones, but the underlying molecular mechanism of ethylene-induced leaf senescence remains poorly understood. RESULTS: In this study, we identified one AT-hook like (AHL) protein, AHL9, as a positive regulator of leaf senescence in Arabidopsis thaliana. Overexpression of AHL9 significantly accelerates age-related leaf senescence and promotes dark-induced leaf chlorosis. The early senescence phenotype observed in AHL9 overexpressing lines is inhibited by the ethylene biosynthesis inhibitor aminooxyacetic acid suggesting the involvement of ethylene in the AHL9-associated senescence. RNA-seq and quantitative reverse transcription PCR (qRT-PCR) data identified numerous senescence-associated genes differentially expressed in leaves of AHL9 overexpressing transgenic plants. CONCLUSIONS: Our investigation demonstrates that AHL9 functions in accelerating the leaf senescence process via ethylene synthesis or signalling.
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Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Etilenos/metabolismo , Regulação da Expressão Gênica de Plantas , Folhas de Planta/genética , Folhas de Planta/metabolismo , Senescência Vegetal , Plantas Geneticamente Modificadas/metabolismo , Fatores de Transcrição/genéticaRESUMO
BACKGROUND: The production of platelets is tightly regulated by thrombopoietin (THPO). Mutations in the THPO gene cause thrombocytopenia. Although mice lacking Thpo present with thrombocytopenia, predicting phenotypes and pathogenicity of novel THPO mutations in mice is limited. Zebrafish can be a powerful tool for fast validation and study of candidate genes of human hematological diseases and have already been used as a model of human thrombocytopenia. OBJECTIVES: We aim to investigate the role of Thpo in zebrafish thrombopoiesis and to establish a Thpo-deficient zebrafish model. The model could be applied for illustrating the clinically discovered human THPO variants of which the clinical significance is not known and to evaluate the effect of THPO receptor agonists (THPO-Ras), as well as a screening platform for new drugs. METHODS: We generated a thpo loss-of-function zebrafish model using CRISPR/Cas9. After disruption of zebrafish thpo, thposzy6 zebrafish presented with a significant reduction of thpo expression and developed thrombocytopenia. Furthermore, we performed in vivo studies with zebrafish with the thposzy6 mutation and found two human clinical point mutations (c.091C > T and c.112C > T) that were responsible for the thrombocytopenia phenotype. In addition, effects of THPO-RAs used as therapeutics against thrombocytopenia were evaluated in the Tg(mpl:eGFP);thposzy6 line. RESULTS AND CONCLUSIONS: Zebrafish with the mutation thposzy6 presented with a significant reduction of thpo expression and developed thrombocytopenia. Thpo loss-of-function zebrafish model can serve as a valuable preclinical model for thrombocytopenia caused by thpo-deficiency, as well as a tool to study human clinical THPO variants and evaluate the effect of THPO-RAs.
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Trombocitopenia , Trombopoetina , Animais , Modelos Animais de Doenças , Humanos , Receptores de Trombopoetina/genética , Receptores de Trombopoetina/metabolismo , Trombocitopenia/genética , Trombocitopenia/metabolismo , Trombopoese/genética , Trombopoetina/genética , Trombopoetina/metabolismo , Peixe-Zebra/genética , Peixe-Zebra/metabolismoRESUMO
Chlamydia trachomatis (C. trachomatis) is a kind of intracellular parasitic microorganism, which can causes many diseases such as trachoma. In this strategy, a specific hairpin DNA with the probe loop as specific regions to recognize C. trachomatis DNA with strong affinity was designed, and its stem consisted of 24 AT base pairs as an effective template for hairpin DNA-CuNCs formation. In the absence of C. trachomatis DNA, the detection system showed strong orange fluorescence emission peaks at 606 nm. In the presence of C. trachomatis DNA, the conformation of DNA probe changed after hybridizing with C. trachomatis DNA. Then, the amount of hairpin DNA-CuNCs was reduced and resulted in low fluorescence emission. C. trachomatis DNA displayed a significant inhibitory effect on the synthesis of fluorescent hairpin DNA-CuNCs due to the competition between C. trachomatis DNA and the specific hairpin DNA. Under the optimal experimental conditions, different concentrations of C. trachomatis were tested and the results showed a good linear relationship in the range of 50 nM to 950 nM. Moreover, the detection limit was 18.5 nM and this detection method possessed good selectivity. Finally, the fluorescent biosensor had been successfully applied to the detection of C. trachomatis target sequence in HeLa cell lysate, providing a new strategy for the detection of C. trachomatis.
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Técnicas Biossensoriais , Nanopartículas Metálicas , Chlamydia trachomatis/genética , Cobre , DNA , Corantes Fluorescentes , Células HeLa , Humanos , Limite de Detecção , Espectrometria de Fluorescência/métodosRESUMO
One of the most important vector-borne disease in humans is malaria, caused by Plasmodium parasite. Seasonal temperature elements have a major effect on the life development of mosquitoes and the development of parasites. In this paper, we establish and analyze a reaction-diffusion model, which includes seasonality, vector-bias, temperature-dependent extrinsic incubation period (EIP) and maturation delay in mosquitoes. In order to get the model threshold dynamics, a threshold parameter, the basic reproduction number $ R_{0} $ is introduced, which is the spectral radius of the next generation operator. Quantitative analysis indicates that when $ R_{0} < 1 $, there is a globally attractive disease-free $ \omega $-periodic solution; disease is uniformly persistent in humans and mosquitoes if $ R_{0} > 1 $. Numerical simulations verify the results of the theoretical analysis and discuss the effects of diffusion and seasonality. We study the relationship between the parameters in the model and $ R_{0} $. More importantly, how to allocate medical resources to reduce the spread of disease is explored through numerical simulations. Last but not least, we discover that when studying malaria transmission, ignoring vector-bias or assuming that the maturity period is not affected by temperature, the risk of disease transmission will be underestimate.
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Malária , Plasmodium , Animais , Número Básico de Reprodução , Humanos , Modelos Biológicos , Mosquitos VetoresRESUMO
The effects of condiment (salt) and processing technic (homogenization) on digestion and interfacial properties of micro/nano-sized colloidal particles (MNCPs) in bigeye tuna head soup (BTHS) using simulated gastrointestinal digestion model in vitro were investigated. For MNCPs in BTHS, the triglycerides were wrapped with proteins in the form of a ring. After salting, the average particle size of the MNCPs in salted BTHS (SBTHS) decreased compared with BTHS. However, the partial demulsification phenomenon existed, and part of the protein was encapsulated in some MNCPs. After further homogenization, the average particle size of the MNCPs in homogenous SBTHS (HSBTHS) was further decreased based on SBTHS and the MNCP was rearranged, which changed the original membrane structure. After gastrointestinal digestion, adding salt decreased the release of total fatty acids compared with unsalted. But homogenization processing increased the release of total fatty acids in HSBTHS and there was no significant difference (p ≥ 0.05) between HSBTHS and BTHS. Thus, the decrease in the release of some fatty acids due to adding salt was compensated by homogenization. Therefore, the changes in composition and microstructure of MNCPs induced by salt and homogenization might contribute to the digestion difference of MNCPs.
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INTRODUCTION: Berberine was one of the active components in Chinese herb and exerted tumor suppressive role in cancer progression, but the exact antitumor mechanism is still not clearly clarified. In the present study, bioinformatics analysis was performed on COAD patients from TCGA, HPA database, UALCAN and GEPIA 2 platform. We also explored the role of berberine on progression of human colon cancers in vitro and in vivo and clarified weather the antitumor effects of berberine was mediated by Wnt/beta-catenin pathway. METHODS: Cell viability was determined by MTT assay. The protein levels were tested by western blotting and the distribution of ß-catenin was observed by confocal microscope. RESULTS: The results showed the levels of CTNNB1 mRNA was increased in colon cancer patients than normal controls. The diagnostic value of CTNNB1 was AUC = 0.882 (CI:0.854-0.911) with sensitivity of 1.000 and specificity of 0.777. The promoter methylation level of CTNNB1 in COAD patients was significantly decreased. Moreover, univariate analysis and multivariate analysis results showed the expression of CTNNB1 in COAD patients was associated with T stage (p = 0.010), pathological stage (p = 0.025) and perineural invasion (p = 0.025). Furthermore, the in vitro assay results showed ß-catenin signaling was highly activated in human colon cancer cells and berberine inhibited the cell viability of colon cancer cells in vitro and in vivo in a dose-and time-dependent manner. Moreover, berberine induced the translocation of ß-catenin to cytoplasm from nucleus. CONCLUSION: The levels of CTNNB1 mRNA was increased in colon cancer patients than normal controls. Berberine inhibited the proliferation of colon cancer cells by regulating the beta-catenin signaling pathway.
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Berberina/uso terapêutico , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/patologia , Progressão da Doença , Transdução de Sinais , beta Catenina/metabolismo , Idoso , Animais , Berberina/farmacologia , Estudos de Casos e Controles , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/genética , Metilação de DNA/efeitos dos fármacos , Metilação de DNA/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Análise Multivariada , Regiões Promotoras Genéticas/genética , Modelos de Riscos Proporcionais , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fatores de Tempo , beta Catenina/genéticaRESUMO
Postnatal colonization and development of the gut microbiota is linked to health and growth. A comprehensive understanding of the postnatal compositional changes and development of the microbial community is helpful to understand the gut health and improve the survival rate of South China tiger cubs (Panthera tigris amoyensis). Fecal samples from three tiger cubs were collected on the day of birth in 2018 (June 17-21 [G0], July 18 [G1], July 31 [G2], and August 7 [G3]). The 16S rRNA genes of the fecal microflora were sequenced. Results showed that 38 phyla, 58 classes, 134 orders, 272 families, and 636 genera of bacteria from 3,059 operational taxonomic units were identified from 12 fecal samples. The diversity and abundance of species of group G0 were significantly higher (p < 0.05 or 0.01) than those of groups G2 and G3. The predominant phylum was Proteobacteria in groups G0 and G1 (38.85% and 48%, respectively) and Firmicutes in groups G2 and G3 (71.42% and 75.29%, respectively). At the phylum level, the abundance of Deinococcus-Thermus was significantly decreased in groups G1, G2, and G3 as compared to group G0 (p < 0.05), while that of Firmicutes was significantly increased in groups G2 and G3 (p < 0.05). At the genus level, the abundance of Faecalibacterium, Ralstonia, and unidentified Rickettsiales was significantly decreased in groups G1, G2, and G3 as compared with group G0 (p < 0.05), while that of Pseudomonas was significantly decreased in groups G2 and G3 (p < 0.05). The composition and structure of fecal microbiota of South China tiger cubs changed after birth.