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1.
Zhongguo Dang Dai Er Ke Za Zhi ; 24(7): 786-791, 2022 Jul 15.
Artigo em Chinês | MEDLINE | ID: mdl-35894194

RESUMO

OBJECTIVES: To study the clinical value of extracorporeal membrane oxygenation (ECMO) in the treatment of persistent pulmonary hypertension of the newborn (PPHN). METHODS: A retrospective analysis was performed on the medical data of 11 neonates with PPHN who were treated with ECMO in the Neonatal Intensive Care Unit of Zhongshan People's Hospital from January 2015 to December 2021, involving the neonates' general information, clinical diagnosis, laboratory results, duration of ECMO treatment, complications during ECMO treatment, length of hospital stay, and outcome. RESULTS: Of the 11 neonates, 10 (91%) had successful weaning from ECMO, and 8 (73%) survived. For the 11 neonates, the mean duration of ECMO treatment was (81±50) hours (range: 26 to 185 hours), the mean duration of ventilator use was (198±105) hours (range: 57 to 392 hours), and the mean length of hospital stay was (22±15) days (range: 2 to 49 days). The oxygenation index and blood lactate level were significantly improved after 24 hours of ECMO treatment among the 11 neonates (P<0.05). Ten neonates had significantly reduced pulmonary artery pressure after 24 hours of ECMO treatment (P<0.05). One neonate had a progressive increase in the pulmonary artery pressure during EMCO treatment, succumbing to death. This neonate was diagnosed with alveolar capillary dysplasia based on the histopathological findings of the lung tissue and whole-exome sequencing results. Among the 11 children, 5 had intracranial hemorrhage, 1 had disseminated intravascular coagulation, 1 had gastric hemorrhage, 2 had pulmonary hemorrhage, 1 had renal insufficiency, and 3 had bleeding at the puncture site during ECMO treatment. CONCLUSIONS: ECMO is effective for the treatment of PPHN, however, the high incidence of complications of ECMO treatment suggests that it is important to carefully assess the indications and timing of ECMO treatment and improve the management of ECMO, which can improve the weaning rate and survival rate.


Assuntos
Oxigenação por Membrana Extracorpórea , Hipertensão Pulmonar , Pneumopatias , Síndrome da Persistência do Padrão de Circulação Fetal , Criança , Humanos , Hipertensão Pulmonar/terapia , Recém-Nascido , Síndrome da Persistência do Padrão de Circulação Fetal/terapia , Estudos Retrospectivos , Resultado do Tratamento
2.
Diabetes ; 2022 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-35856496

RESUMO

The neuropeptide AgRP is essential for maintaining systemic energy homeostasis. In the present study, we show that hypothalamic Foxi2, as a novel regulator of nutrient sensing, controls systemic energy metabolism by specifically stimulating AgRP expression. Foxi2 was highly expressed in the hypothalamus, and its expression was induced by fasting. Immunofluorescence assays demonstrated that Foxi2 was localized in AgRP neurons. We stereotaxically injected AAV to selectively overexpress Foxi2 in AgRP-IRES-Cre mice and found that Foxi2 overexpression in AgRP neurons specifically increased AgRP expression, thereby increasing food intake and reducing energy expenditure, subsequently leading to obesity and insulin resistance. Mechanistically, Foxi2 stimulated AgRP expression by directly binding to it and activating its transcription. Furthermore, Foxi2 overexpression activated AgRP neuron activity, as revealed by whole-cell patch-clamp experiments. Conversely, global Foxi2 mutant mice became leaner with age and were resistant to high-fat diet (HFD)-induced obesity and metabolic disturbances. Collectively, our data suggest that Foxi2 plays an important role in controlling energy metabolism by regulating AgRP expression.

3.
Sci Rep ; 12(1): 12698, 2022 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-35882972

RESUMO

In order to explore the influence of weak interlayer on blasting characteristics in natural rock mass, by using the particle flow code (PFC2D), a single hole blasting numerical model of hard rock with soft interlayer is established. The blasting experiments at different positions and thicknesses of weak interlayer are carried out. Then an in-depth analysis from the perspectives of crack effect, stress field and energy field is made. Results showed that: (i) When the explosion is initiated outside the weak interlayer, if the interlayer is located within about twice the radius of the crushing area, the closer the interlayer is to the blast hole, the higher the damage degree of the rock mass around the blast hole. And the number of cracks will increase by about 1-2% when the distance between the weak interlayer and the blast hole decreases by 0.5 m. (ii) When detonating outside the weak interlayer, if the interlayer is within about 4 times radius of the crushing area, the hard rock on both sides of the weak interlayer will in a high stress state. Under the same case, the peak kinetic energy and peak friction energy will increase by about 23 and 13%, respectively, and the peak strain energy will increase by about 218 kJ for every 0.1 m increase in the thickness of the weak interlayer.

4.
Int J Gen Med ; 15: 5455-5463, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35692354

RESUMO

Objective: To investigate computed tomography (CT) features of stage IA invasive mucinous adenocarcinoma (IMA) of the lung and establish a predictive model. Methods: Fifty-three lesions from 53 cases of stage IA IMA between January 2017 and December 2019 were examined, while 141 lesions from 141 cases of invasive non-mucinous lung adenocarcinoma (INMA) served as control cases. Univariate analysis was performed to compare differences in demographics and CT features between the two groups, and multivariate logistic regression analysis was performed to determine primary influencing factors of solitary nodular IMA. A risk score prediction model was established based on the regression coefficients of these factors, and receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive performance of the model. Results: Univariate analysis showed that age, nodule type, maximum nodule diameter, tumor lung interface, lobulation, spiculation, air bronchogram or vacuolar signs, and abnormal vascular changes differed significantly between the two groups (p < 0.05). Compared to INMA, spiculation of IMA was relatively longer and softer. Multivariate logistic regression analysis showed that nodule type, indistinct tumor lung interface, air bronchogram or vacuolar signs, and abnormal vascular changes were the primary influencing factors. A prediction model based on the regression coefficients of these factors was established. ROC curve analysis indicated that the area under the curve was 0.882 (p < 0.05). Conclusion: Compared to INMA, solitary peripheral stage IA nodular IMA were more common in older patients; they more frequently had indistinct tumor lung interface and air bronchogram or vacuolar signs on CT; spiculation was relatively longer and softer; our risk score prediction model based on nodule type, tumor lung interface, air bronchogram or vacuolar signs, and abnormal vascular changes was established with good predictive efficacy for solitary nodular IMA.

5.
Curr Med Sci ; 42(3): 555-560, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35678914

RESUMO

Coronavirus disease 2019 (COVID-19) has caused a global pandemic impacting over 200 countries/regions and more than 200 million patients worldwide. Among the infected patients, there is a high prevalence of COVID-19-related cardiovascular injuries. However, the specific mechanisms linking cardiovascular damage and COVID-19 remain unclear. The COVID-19 pandemic also has exacerbated the mental health burden of humans. Considering the close association between neuroimmune interactions and cardiovascular disease, this review assessed the complex pathophysiological mechanisms connecting neuroimmune interactions and cardiovascular disease. It was revealed that the mental health burden might be a pivotal accomplice causing COVID-19-associated cardiovascular damage. Specifically, the proinflammatory status of patients with a terrible mood state is closely related to overdrive of the hypothalamus-pituitary-adrenal (HPA) axis, sympathovagal imbalance, and endothelial dysfunction, which lead to an increased risk of developing cardiovascular injury during COVID-19. Therefore, during the prevention and treatment of cardiovascular complications in COVID-19 patients, particular attention should be given to relieve the mental health burden of these patients.


Assuntos
COVID-19 , Doenças Cardiovasculares , COVID-19/complicações , Humanos , Neuroimunomodulação , Pandemias , SARS-CoV-2
6.
Artigo em Inglês | MEDLINE | ID: mdl-35730192

RESUMO

BACKGROUND AND AIM: The diagnosis and treatment of gastrointestinal (GI) bleeding secondary to malignancy can be challenging. Endoscopy is the gold standard to diagnose and treat gastrointestinal bleeding but clinical characteristics and outcomes of patients with malignancy-related bleeding are not well understood. This study aims to look at clinical characteristics, endoscopic findings, safety and clinical outcomes of endoscopic interventions for GI malignancy-related bleeding. METHODS: We retrospectively reviewed outcomes of patients with confirmed GI malignancies who underwent endoscopy for GI bleeding at MD Anderson Cancer Center between 2010 and 2019. Cox hazard analysis was conducted to identify factors associated with survival. RESULTS: A total of 313 patients were included, with median age of 59 years; 74.8% were male. The stomach (30.0%) was the most common tumor location. Active bleeding was evident endoscopically in 47.3% of patients. Most patients (77.3%) did not receive endoscopic treatment. Of the patients who received endoscopic treatment, 57.7% had hemostasis. No endoscopy-related adverse events were recorded. Endoscopic treatment was associated with hemostasis (P < 0.001), but not decreased recurrent bleeding or mortality. Absence of active bleeding on endoscopy, stable hemodynamic status at presentation, lower cancer stage, and surgical intervention were associated with improved survival. CONCLUSIONS: This study indicates that endoscopy is a safe diagnostic tool in this patient population; while endoscopic treatments may help achieve hemostasis, it may not decrease the risk of recurrent bleeding or improve survival.

7.
ESC Heart Fail ; 2022 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-35736641

RESUMO

AIMS: We aim to investigate the additive effect of B-lines on lung ultrasound (LUS) for predicting outcome in patients with heart failure (HF) when combined with conventional assessment of clinical congestion. METHODS AND RESULTS: This study prospectively enrolled 117 hospitalized HF patients (61 ± 16 years, 70.1% males) who underwent congestion assessment by the 'wet/dry' status, clinical congestion score (CCS), and B-lines on LUS. The primary endpoint was all-cause mortality or hospitalization for HF during the 180-day follow-up after discharge. The 'Wet', CCS ≥ 3, and B-lines >5, indicators of congestion positive (+), were observed in 83.8%, 76.1%, and 70.1% of the patients on admission, respectively; and the numbers significantly decreased to 41.9%, 41.9%, and 35.9% at discharge, respectively. The agreement between the 'wet/dry' status and B-lines (58.1%) or between CCS and B-lines (56.4%) was moderate at discharge, in terms of both positive and both negative. By incorporating the B-lines with assessment of clinical congestion, the patients at discharge were divided into three phenotypes as clinical congestion (+), clinical congestion (-) with B-lines (+), and clinical congestion (-) with B-lines (-). The Kaplan-Meier analysis showed a better survival in the both (-) group ('wet/dry' with B-lines: Chi-square 10.591, P = 0.005; CCS with B-lines: χ2 6.239, P = 0.031). When the 'wet' patients (n = 49) being taken as the reference, the 'dry' patients with B-lines (+) (n = 21) had an identical risk of the composite endpoint (hazard ratio [HR] adjusted for clinical covariates 1.021, 95% confidence interval [CI] 0.480-2.134, P = 0.974), while the 'dry' patients with B-lines (-) (n = 47) had a lower risk (HR 0.264, 95% CI 0.113-0.617, P = 0.002). When the CCS (+) patients (n = 49) being regarded as the reference, similar results were obtained in the patients with CCS (-) but B-lines (+) (n = 22) (HR 1.348, 95% CI 0.627-2.896, P = 0.444) as well as in those with both CCS (-) and B-lines (-) (n = 46) (HR 0.447, 95% CI 0.202-0.992, P = 0.048). CONCLUSIONS: The combination of B-lines on LUS and conventional assessment helped to identify new phenotypes of congestion that aid in the risk stratification of discharged HF patients. Further investigation is warranted to determine whether this strategy could be adopted as a guide for decongestion therapy.

9.
Nat Commun ; 13(1): 2539, 2022 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-35534483

RESUMO

Extrapulmonary complications of different organ systems have been increasingly recognized in patients with severe or chronic Coronavirus Disease 2019 (COVID-19). However, limited information on the skeletal complications of COVID-19 is known, even though inflammatory diseases of the respiratory tract have been known to perturb bone metabolism and cause pathological bone loss. In this study, we characterize the effects of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on bone metabolism in an established golden Syrian hamster model for COVID-19. SARS-CoV-2 causes significant multifocal loss of bone trabeculae in the long bones and lumbar vertebrae of all infected hamsters. Moreover, we show that the bone loss is associated with SARS-CoV-2-induced cytokine dysregulation, as the circulating pro-inflammatory cytokines not only upregulate osteoclastic differentiation in bone tissues, but also trigger an amplified pro-inflammatory cascade in the skeletal tissues to augment their pro-osteoclastogenesis effect. Our findings suggest that pathological bone loss may be a neglected complication which warrants more extensive investigations during the long-term follow-up of COVID-19 patients. The benefits of potential prophylactic and therapeutic interventions against pathological bone loss should be further evaluated.


Assuntos
COVID-19 , Animais , COVID-19/complicações , Cricetinae , Modelos Animais de Doenças , Humanos , Mesocricetus , SARS-CoV-2
10.
Chem Commun (Camb) ; 58(40): 6024-6027, 2022 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-35502741

RESUMO

Durable Cu/NiFe(OH)x electrocatalyst was designed for hydrogen evolution reaction in alkaline media. The in situ generated Cu nanodendrites protect the NiFe(OH)x from being hydrogenated, giving it a > 1000 h lifetime for high-performance water splitting (1.51 V, 10 mA cm-2 in 1 M KOH) when coupled with a NiFe-layered double hydroxide anode.

11.
Clin Cancer Res ; 28(9): 1938-1947, 2022 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-35491652

RESUMO

PURPOSE: Intraductal papillary mucinous neoplasms (IPMN) are bona fide precursors to pancreatic ductal adenocarcinoma (PDAC). While genomic alterations during multistep IPMN progression have been well cataloged, the accompanying changes within the tumor immune microenvironment (TIME) have not been comprehensively studied. Herein, we investigated TIME-related alterations during IPMN progression, using multiplex immunofluorescence (mIF) coupled with high-resolution image analyses. EXPERIMENTAL DESIGN: Two sets of formalin-fixed, paraffin-embedded tissue samples from surgically resected IPMNs were analyzed. The training set of 30 samples consisted of 11 low-grade IPMN (LG-IPMN), 17 high-grade IPMN (HG-IPMN), and 2 IPMN with PDAC, while a validation set of 93 samples comprised of 55 LG-IPMN and 38 HG-IPMN. The training set was analyzed with two panels of immuno-oncology-related biomarkers, while the validation set was analyzed with a subset of markers found significantly altered in the training set. RESULTS: Cell types indicative of enhanced immune surveillance, including cytotoxic and memory T cells, and antigen-experienced T cells and B cells, were all found at higher densities within isolated LG-IPMNs compared with HG-IPMNs. Notably, the TIME of LG-IPMNs that had progressed at the time of surgical resection (progressor LGD) resembled that of the synchronous HG-IPMNs, underscoring that attenuated immune surveillance occurs even in LG-IPMNs destined for progression. CONCLUSIONS: Our findings provide a basis for interception of cystic neoplasia to PDAC, through maintenance of sustained immune surveillance using vaccines and other prevention approaches.


Assuntos
Adenocarcinoma Mucinoso , Carcinoma Ductal Pancreático , Neoplasias Intraductais Pancreáticas , Neoplasias Pancreáticas , Adenocarcinoma Mucinoso/patologia , Carcinoma Ductal Pancreático/patologia , Humanos , Neoplasias Pancreáticas/patologia , Microambiente Tumoral
12.
Biomater Adv ; 134: 112558, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35525754

RESUMO

Cancer treatment is imminent, and controlled drug carriers are an important development direction for future clinical chemotherapy. Visual guidance is a feasible means to achieve precise treatment, reduce toxicity and increase drug efficacy. However, the existing visual control methods are limited by imaging time-consuming, sensitivity and side effects. In addition, the ability of the carrier to respond to environmental stimuli in vivo is another difficulty that limits its application. Here, we propose a highly stimulus-responsive GC liposome with precise tracing and sensitive feedback capabilities. It combines magnetic resonance imaging and fluorescence imaging, and addresses the need for precise visualization by alternating imaging modalities. More importantly, GC liposomes are a carrier that can accumulate stimuli. In this paper, by tracking the fragmentation process of empty GC and drug-loaded D-GC liposomes, we confirm the synergistic effect between multiple stimuli, which can result in a more efficient drug release performance. Finally, in mice models we examined the GC liposome imaging approach and the D-GC + UV group guided by this visualization exhibited the highest tumor inhibition efficiency (6.85-fold). This study highlights the advantages of alternate visualization-guided and co-stimulation treatment strategies, and provides design ideas and potential materials for efficient and less toxic cancer treatments.


Assuntos
Lipossomos , Neoplasias , Animais , Portadores de Fármacos , Liberação Controlada de Fármacos , Imageamento por Ressonância Magnética/métodos , Camundongos
13.
Artigo em Inglês | MEDLINE | ID: mdl-35469157

RESUMO

Context. Ziziphi Spinosae Semen (ZSS) is a well-known Chinese herbal medicine used in the treatment of depression and anxiety in China. ZSS contains several active components, such as alkaloids, saponins, and flavonoids. Objective. This study aimed to explore the synergistic effect of alkaloids and saponins from ZSS in alleviating depression in a mouse model. Materials and Methods. Modeling depression with chronic unpredictable stimuli. Pharmacodynamic methods (tail suspension test and forced swimming test) were used to evaluate the antidepressant effects of alkaloids, saponins, and combinations thereof from ZSS. The mechanisms underlying the effect were examined by measuring the levels of monoamine neurotransmitters in the hippocampus and frontal cortex of mice. Results. Compared with the model group, alkaloids therapy (AZSS), saponins therapy (SZSS), and combination therapy significantly reduced the immobility time in behavioral tests (P < 0.05). The contents of noradrenaline (NE), dopamine (DA), and serotonin (5-HT) in the hippocampus and frontal cortex of depressed mice were increased in the drug treatment groups, especially in the combination group, which increased by 14.65%, 87.72%, 33.44%, 25.64%, 25.39%, and 70.78%, respectively. Several groups showed better results (P < 0.05), especially the combination of alkaloids and saponins. Discussion and Conclusion. The saponins and alkaloids from ZSS exhibited a synergistic effect in improving the behavior of depressed mice. More importantly, the combination of alkaloids (15 mg·kg-1) and saponins (110 mg·kg-1) was effective in alleviating depression in mice, especially in terms of changing the level of DA in the hippocampus.

14.
Sci Total Environ ; 834: 155321, 2022 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-35452730

RESUMO

Livestock waste is a known reservoir of Escherichia coli (E. coli) carrying clinically important CTX-M-type extended-spectrum ß-lactamase genes (blaCTX-M), however, the occurrence and transfer characteristics of blaCTX-M genes during anaerobic digestion (AD) remain unclear. Herein, four full-scale and two parallel lab-scale AD systems treating swine waste under ambient and mesophilic conditions were investigated by both molecular- and culture-based methods to reveal the occurrence and transfer behaviors of blaCTX-M genes during AD. Real-time TaqMan polymerase chain reaction revealed 1.3 × 104-6.8 × 105 and 3.0 × 104-7.0 × 105 copies/mL of blaCTX-M groups 1 and 9 in all feeding substrates. While AD reduced the absolute abundance of groups 1 and 9 by 0.63-2.24 and 0.08-1.30 log (P < 0.05), 5.0 × 102-4.1 × 103 and 1.1 × 104-3.5 × 104 copies/mL of groups 1 and 9 remained in the anaerobic effluent, respectively. In total, 141 blaCTX-M-carrying E. coli isolates resistant to cefotaxime were obtained from the AD reactors. Whole-genome sequencing showed that blaCTX-M-65 mainly carried by E. coli ST155 was the most frequently detected group 9 subtype in the feeding substrate; whereas blaCTX-M-14 associated with the dominant clones E. coli ST6802 and ST155 became the major subtype in AD effluent. Furthermore, blaCTX-M-14 was flanked by ΔIS26 upstream and ΔIS903B downstream. The ΔIS26-blaCTX-M-14-ΔIS903B element was mainly located on the IncHI2 plasmid in E. coli ST48 and ST6802 and also the IncFIB plasmid in ST155 in anaerobic effluent. Conjugation assays showed that the plasmids harboring blaCTX-M-14 could be successfully transferred at a frequency of 10-3-10-2 cells per recipient cell. This study revealed that blaCTX-M genes remained in both the full-scale and lab-scale AD effluents of swine waste. Thus, additional efforts should be implemented to block the discharge and spread of antibiotic resistance genes to the environment.


Assuntos
Infecções por Escherichia coli , Escherichia coli , Anaerobiose , Animais , Antibacterianos , Clonidina , Escherichia coli/genética , Escherichia coli/metabolismo , Infecções por Escherichia coli/epidemiologia , Plasmídeos , Suínos , beta-Lactamases/genética , beta-Lactamases/metabolismo
15.
NPJ Breast Cancer ; 8(1): 46, 2022 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-35393462

RESUMO

Patients with hereditary mutations in BRCA1 or BRCA2 (gBRCA1/2) and breast cancer have distinct tumor biology, and encompass a predilection for brain metastasis (BM). We looked into baseline risk of BMs among gBRCA1/2 patients. Patients with gBRCA1/2, stage I-III invasive breast cancer seen between 2000-2017 with parenchymal BMs. Among gBRCA1 with distant breast cancer recurrence, 34 of 76 (44.7%) were diagnosed with brain metastases compared to 7 of 42 (16.7%) patients with gBRCA2. In the comparator group, 65 of 182 (35.7%) noncarrier triple-negative breast cancer (TNBC) and a distant recurrence experienced BM's. In a competitive risk analysis using death as a competing factor, the cumulative incidence of BMs was similar between gBRCA1 and noncarrier TNBC patients. The time from primary breast cancer diagnosis to detection of BMs was similar between gBRCA1 and noncarrier TNBC patients (2.4 vs 2.2 years). Survival was poor after BMs (7.8 months for gBRCA1 patients vs. 6.2 months for TNBC noncarriers). Brain was a more common site of initial distant recurrence in gBRCA1 patients versus TNBC noncarriers (26.3% vs. 12.1%). Importantly, the presence of BMs, adversely impacted overall survival across groups (HR 1.68 (95% CI 1.12-2.53), hazard ratio for death if a patient had BMs at the time of initial breast cancer recurrence vs. not). In conclusion, breast cancer BMs is common and is similarly frequent among gBRCA1 and noncarrier patients with recurrent TNBC. Our study highlights the importance of improving the prevention and treatment of BMs in patients with TNBC, gBRCA1 carriers, and noncarriers.

16.
Bioresour Technol ; 354: 127205, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35462015

RESUMO

This study investigated and assessed the effect of the functional-membrane covering technique (FMCT) on nitrogen succession during aerobic composting. By comparative experiments involving high-throughput sequencing and qPCR, nitrogen metabolism (including the ko00910 pathway and functional enzyme and gene abundances) was analyzed, and the nitrogen succession mechanism was identified. The FMCT created a micro-positive pressure, improved the aerobic conditions, and increased the oxygen utilization rate and temperature. This strongly affected the nitrogen metabolism pathway and down-regulated the nitrifying and denitrifying bacteria abundances. The FMCT up-regulated the relative abundance of glutamate dehydrogenase and down-regulated the absolute abundances of AOB and nxrA. This and the high temperature increased NH3 emissions by 13.78%-73.37%. The FMCT down-regulated the abundances of denitrifying gene groups (nirS + nirK)/nosZ and nitric oxide reductase associated with N2O emissions and decreased N2O emissions by 16.44%-41.15%. The results improve the understanding of the mechanism involved in nitrogen succession using the FMCT.


Assuntos
Compostagem , Desnitrificação , Redes e Vias Metabólicas , Nitrogênio/análise , Óxido Nitroso/análise , Solo , Microbiologia do Solo
17.
Methods Mol Biol ; 2464: 49-64, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35258824

RESUMO

Protoplast transfection is widely used in plant research to rapidly evaluate RNA degradation, reporter assay, gene expression, subcellular localization, and protein-protein interactions. In order to successfully use protoplast transfection with the newly emerging clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) protein editing platform, high yield of protoplasts, stable transfection efficiency, and reliable regeneration protocols are necessary. The Nicotiana tabacum transient protoplast transfection and regeneration system can effectively obtain target gene mutations in regenerated plants without transgenes and is thus a very attractive technique for evaluating gene editing reagents using CRISPR/Cas-based systems. Here, we describe in detail sterilized seed germination, culture conditions, isolation of Nicotiana tabacum protoplasts from tissue culture explants, construction of a vector containing the Cas protein and sgRNA cassette, highly efficient polyethylene glycol-calcium transient transfection of plasmids delivered into protoplasts, evaluation of mutagenesis efficiency and genotype analysis from protoplasts and regenerated plants, and the regeneration conditions to obtain CRISPR-edited plants from single protoplasts.


Assuntos
Sistemas CRISPR-Cas , Protoplastos , Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Mutagênese , Protoplastos/metabolismo , Tabaco/genética
18.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 44(1): 164-172, 2022 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-35300780

RESUMO

Mechanical stimulus is critical to cardiovascular development during embryogenesis period.The mechanoreceptors of endocardial cells and cardiac myocytes may sense mechanical signals and initiate signal transduction that induce gene expression at a cellular level,and then translate molecular-level events into tissue-level deformations,thus guiding embryo development.This review summarizes the regulatory roles of mechanical signals in the early cardiac development including the formation of heart tube,looping,valve and septal morphogenesis,ventricular development and maturation.Further,we discuss the potential mechanical transduction mechanisms of platelet endothelial cell adhesion molecule 1-vascular endothelial-cadherin-vascular endothelial growth factor receptor 2 complex,primary cilia,ion channels,and other mechanical sensors that affect some cardiac malformations.


Assuntos
Coração , Mecanotransdução Celular , Animais , Coração/embriologia , Humanos , Miócitos Cardíacos/fisiologia , Fator A de Crescimento do Endotélio Vascular/metabolismo
19.
Phytomedicine ; 100: 154055, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35344716

RESUMO

BACKGROUND: Dahuang Zhechong pill (DHZCP) improves the inhibitory immune status of mice with hepatocellular carcinoma (HCC) by regulating Treg/Th1 balance. HYPOTHESIS/PURPOSE: To study the multi-material basis and multi-mechanisms of DHZCP against HCC by regulating Treg/Th1 balance in vitro and in vivo. METHODS: UPLC-MS/MS was used to detect the dynamic changes in 29 characteristic components of different polar parts of DHZCP. H&E and TUNEL were used to check pathological condition in HCC mice. The number of CD4+T, CD8+T, Treg, Th1, and Th1-like Treg cells was counted by flow cytometry. TGF-ß, IL-10, IFN-γ, and TNF-α content were detected by ELISA. α-Ketoglutarate and glutamine levels were detected by Trace1310/TSQ8000 GC-MS/MS. p-Smad2, and p-Smad3 protein levels were detected by WB, mRNA expression of Smad2, alanine-serine-cysteine transporter-2, glutaminase, and glutamate dehydrogenase were detected by RT-PCR. Simca-p multivariate data analysis software was used to evaluate the relationship between the different polar parts of DHZCP and the proportion of Treg cells. RESULTS: Water-soluble (PW) and ethyl acetate (PE) polar parts of DHZCP affected the HCC immune system by inhibiting the differentiation of Tregs, reversing the balance of Treg/Th1, and significantly reduced the tumor volume and weight. However, petroleum ether and n-butanol polar parts had no above actions. The changes in emodin, chrysophanol, aloe vera emodin, emodin-8-O-ß-D-glycoside, gallic acid, naringenin, baicalein, wogonin, norwogonin, apigenin, chrysin, glycyrrhizin, formononetin, and palmitic acid were closely related to the changes of Treg cells, which is the main material basis of DHZCP inhibition of Treg differentiation. Additionally, PW mainly inhibit the differentiation of Treg cells by affecting the metabolism of hepatoma cells, improving tumor microenvironment acidity, and glutamine depletion. However, PE inhibited the differentiation of Treg cells mainly by regulating the TGF-ß/Smad pathway. CONCLUSION: In this study, accurate analysis of multi-component was combined with pharmacodynamic evaluations to identify the pharmacodynamic substances of DHZCP in regulating Treg/Th1 balance, and clarified the multi-target mechanism of DHZCP to improve tumor immunity. The study style offers a novel approach for pharmacological research on TCM.


Assuntos
Carcinoma Hepatocelular , Emodina , Neoplasias Hepáticas , Animais , Carcinoma Hepatocelular/tratamento farmacológico , Medicamentos de Ervas Chinesas , Glutamina , Neoplasias Hepáticas/tratamento farmacológico , Camundongos , Linfócitos T Reguladores , Espectrometria de Massas em Tandem , Fator de Crescimento Transformador beta , Microambiente Tumoral
20.
Angew Chem Int Ed Engl ; 61(24): e202204256, 2022 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-35334135

RESUMO

Employing pure water, the ultimate green source of hydrogen donor to initiate chemical reactions that involve a hydrogen atom transfer (HAT) step is fascinating but challenging due to its large H-O bond dissociation energy (BDEH-O =5.1 eV). Many approaches have been explored to stimulate water for hydrogenative reactions, but the efficiency and productivity still require significant enhancement. Here, we show that the surface hydroxylated graphitic carbon nitride (gCN-OH) only requires 2.25 eV to activate H-O bonds in water, enabling abstraction of hydrogen atoms via dehydrogenation of pure water into hydrogen peroxide under visible light irradiation. The gCN-OH presents a stable catalytic performance for hydrogenative N-N coupling, pinacol-type coupling and dehalogenative C-C coupling, all with high yield and efficiency, even under solar radiation, featuring extensive impacts in using renewable energy for a cleaner process in dye, electronic, and pharmaceutical industries.

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