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1.
Environ Res ; 203: 111873, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34411548

RESUMO

A series of rare earth complexes containing (α-PW12O40)3- and PO ligand are synthesized by water bath in 70 °C, [Ln(OPPh3)4(H2O)3](PW12O40)·4CH3CN (Ln = La, Pr, Nd, Sm, Gd, Tb, Ho 1-7) (OPPh3 = Triphenylphosphine oxide, {PW12} = phosphotungstic acid). The precise structures are confirmed by X-ray single crystal diffraction and the result shows all complexes are isostructural. Complexes 1-7 are fully characterized by PXRD, FT-IR, TGA, UV diffuse reflectance spectra and terahertz time-domain spectroscopy (THz-TDS). Complex 3 exhibits the highest photocatalytic degradation efficiency for methylene blue (MB) in this series of complexes. The experimental results showed that the photodegradation efficiency can remain constant at the level of 95% after five consecutive cycles. The photocatalytic reaction kinetics and mechanism of complexes were investigated. Additionally, complexes also exhibit photocatalytic hydrogen evolution activity. THz-TDS was used to characterize the complexes and its raw materials, the characteristic peaks of OPPh3 (broad peak at 1.20 THz) and phosphotungstic acid (sharp peaks at 0.23, 0.32 THz) were obtained.

2.
Adv Sci (Weinh) ; : e2101923, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34719887

RESUMO

Diffuse invasion is the primary cause of treatment failure of glioblastoma (GBM). Previous studies on GBM invasion have long been forced to use the resected tumor mass cells. Here, a strategy to reliably isolate matching pairs of invasive (GBMINV ) and tumor core (GBMTC ) cells from the brains of 6 highly invasive patient-derived orthotopic models is described. Direct comparison of these GBMINV and GBMTC cells reveals a significantly elevated invasion capacity in GBMINV cells, detects 23/768 miRNAs over-expressed in the GBMINV cells (miRNAINV ) and 22/768 in the GBMTC cells (miRNATC ), respectively. Silencing the top 3 miRNAsINV (miR-126, miR-369-5p, miR-487b) successfully blocks invasion of GBMINV cells in vitro and in mouse brains. Integrated analysis with mRNA expression identifies miRNAINV target genes and discovers KCNA1 as the sole common computational target gene of which 3 inhibitors significantly suppress invasion in vitro. Furthermore, in vivo treatment with 4-aminopyridine (4-AP) effectively eliminates GBM invasion and significantly prolongs animal survival times (P = 0.035). The results highlight the power of spatial dissection of functionally accurate GBMINV and GBMTC cells in identifying novel drivers of GBM invasion and provide strong rationale to support the use of biologically accurate starting materials in understanding cancer invasion and metastasis.

3.
JACS Au ; 1(10): 1631-1638, 2021 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-34723266

RESUMO

The structure of the [Rh2C3]+ ion and its reaction with CH4 in the gas phase have been studied by infrared photodissociation spectroscopy and mass spectrometry in conjunction with quantum chemical calculations. The [Rh2C3]+ ion is characterized to have an unsymmetrical linear [Rh-C-C-C-Rh]+ structure existing in two nearly isoenergetic spin states. The [Rh2C3]+ ion reacts with CH4 at room temperature to form [Rh2C]+ + C3H4 and [Rh2C2H2]+ + C2H2 as the major products. In addition to the [Rh2C]+ ion, the [Rh2 13C]+ ion is formed at about one-half of the [Rh2C]+ intensity when the isotopic-labeled 13CH4 sample is used. The production of [Rh2 13C]+ indicates that the linear C3 moiety of [Rh2C3]+ can be replaced by the bare carbon atom of methane with all four C-H bonds being activated. The calculations suggest that the overall reactions are thermodynamically exothermic, and that the two Rh centers are the reactive sites for C-H bond activation and hydrogen atom transfer reactions.

4.
Comput Math Methods Med ; 2021: 9548312, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34745329

RESUMO

Objective: To explore the image enhancement model based on deep learning on the effect of ureteroscopy with double J tube placement and drainage on ureteral stones during pregnancy. We compare the clinical effect of ureteroscopy with double J tube placement on pregnancy complicated with ureteral stones and use medical imaging to diagnose the patient's condition and design a treatment plan. Methods: The image enhancement model is constructed using deep learning and implemented for quality improvement in terms of image clarity. In the way, the relationship of the media transmittance and the image with blurring artifacts was established, and the model can estimate the ureteral stone predicted map of each region. Firstly, we proposed the evolution-based detail enhancement method. Then, the feature extraction network is used to capture blurring artifact-related features. Finally, the regression subnetwork is used to predict the media transmittance in the local area. Eighty pregnant patients with ureteral calculi treated in our hospital were selected as the research object and were divided into a test group and a control group according to the random number table method, 40 cases in each group. The test group underwent ureteroscopy double J tube placement, and the control group underwent ureteroscopy lithotripsy. Combined with the ultrasound scan results of the patients before and after the operation, the operation time, time to get out of bed, and hospitalization time of the two groups of patients were compared. The operation success rate and the incidence of complications within 1 month after surgery were counted in the two groups of patients. Results: We are able to improve the quality of the images prior to medical diagnosis. The total effective rate of the observation group was 100.0%, which is higher than that of the control group (90.0%). The difference between the two groups was statistically significant (P < 0.05). The adverse reaction rate in the observation group was 5.0%, which was lower than 17.5% in the control group. The difference between the two groups was statistically significant (P < 0.05). The comparison results are then prepared. Conclusions: The image enhancement model based on deep learning is able to improve medical diagnosis which can assist radiologists to better locate the ureteral stones. Based on our method, double J tube placement under ureteroscopy has a significant effect on the treatment of ureteral stones during pregnancy, and it has good safety and is worthy of widespread application.

5.
Lab Invest ; 2021 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-34802040

RESUMO

Brain tumors are the leading cause of cancer-related death in children. Tazemetostat is an FDA-approved enhancer of zeste homolog (EZH2) inhibitor. To determine its role in difficult-to-treat pediatric brain tumors, we examined EZH2 levels in a panel of 22 PDOX models and confirmed EZH2 mRNA over-expression in 9 GBM (34.6 ± 12.7-fold) and 11 medulloblastoma models (6.2 ± 1.7 in group 3, 6.0 ± 2.4 in group 4) accompanied by elevated H3K27me3 expression. Therapeutic efficacy was evaluated in 4 models (1 GBM, 2 medulloblastomas and 1 ATRT) via systematically administered tazemetostat (250 and 400 mg/kg, gavaged, twice daily) alone and in combination with cisplatin (5 mg/kg, i.p., twice) and/or radiation (2 Gy/day × 5 days). Compared with the untreated controls, tazemetostat significantly (Pcorrected < 0.05) prolonged survival times in IC-L1115ATRT (101% at 400 mg/kg) and IC-2305GBM (32% at 250 mg/kg, 45% at 400 mg/kg) in a dose-dependent manner. The addition of tazemetostat with radiation was evaluated in 3 models, with only one [IC-1078MB (group 4)] showing a substantial, though not statistically significant, prolongation in survival compared to radiation treatment alone. Combining tazemetostat (250 mg/kg) with cisplatin was not superior to cisplatin alone in any model. Analysis of in vivo drug resistance detected predominance of EZH2-negative cells in the remnant PDOX tumors accompanied by decreased H3K27me2 and H3K27me3 expressions. These data supported the use of tazemetostat in a subset of pediatric brain tumors and suggests that EZH2-negative tumor cells may have caused therapy resistance and should be prioritized for the search of new therapeutic targets.

6.
Nat Commun ; 12(1): 6479, 2021 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-34759281

RESUMO

Metabolic reprogramming evolves during cancer initiation and progression. However, thorough understanding of metabolic evolution from preneoplasia to lung adenocarcinoma (LUAD) is still limited. Here, we perform large-scale targeted metabolomics on resected lesions and plasma obtained from invasive LUAD and its precursors, and decipher the metabolic trajectories from atypical adenomatous hyperplasia (AAH) to adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA) and invasive adenocarcinoma (IAC), revealing that perturbed metabolic pathways emerge early in premalignant lesions. Furthermore, three panels of plasma metabolites are identified as non-invasive predictive biomarkers to distinguish IAC and its precursors with benign diseases. Strikingly, metabolomics clustering defines three metabolic subtypes of IAC patients with distinct clinical characteristics. We identify correlation between aberrant bile acid metabolism in subtype III with poor clinical features and demonstrate dysregulated bile acid metabolism promotes migration of LUAD, which could be exploited as potential targetable vulnerability and for stratifying patients. Collectively, the comprehensive landscape of the metabolic evolution along the development of LUAD will improve early detection and provide impactful therapeutic strategies.

7.
Am J Transl Res ; 13(10): 11548-11555, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34786081

RESUMO

BACKGROUND: Breast carcinoma (BC) is a commonly seen malignancy in women. Although traditional radical mastectomy can improve the survival of patients, it can cause breast loss and chest wall deformities, which seriously affects the daily life of patients and causes anxiety and depression. The purpose of this research project is to investigate the effect of breast reconstruction with latissimus dorsi myocutaneous flap (LDMF) after nipple- and areola-sparing modified radical mastectomy (MRM) on the psychological mood and quality of life (QoL) of patients with stage I BC. METHODS: A total of 102 patients with BC (research group, RG) treated in the Shanghai Fifth People's Hospital, Fudan University from January 2018 to December 2020 were selected for phase I breast reconstruction with LDMF after nipple- and areola-sparing MRM. Concurrently, 50 BC patients (control group, CG) who underwent traditional total mastectomy in our hospital were collected. The activities of daily living (ADL), self-rating anxiety scale (SAS) and self-rating depression scale (SDS) scores were observed before and 1 month after treatment. The intraoperative indicators, postoperative complications, postoperative satisfaction rate and overall survival rate were compared. RESULTS: The Functional Assessment of Cancer Therapy-Breast Cancer (FACT-B) score was higher after treatment, while SAS and SDS scores were lower in RG than in CG (P<0.05). No statistical difference was observed in intraoperative blood loss, wound drainage time, operation time, postoperative complications and overall survival rate between the two cohorts (P>0.05). RG showed higher satisfaction degree and overall satisfaction rate, as well as better QoL than CG (P<0.05). CONCLUSIONS: Breast reconstruction with LDMF after nipple- and areola-sparing MRM can alleviate adverse emotions of patients with stage I BC and improve their QoL.

8.
Huan Jing Ke Xue ; 42(11): 5264-5274, 2021 Nov 08.
Artigo em Chinês | MEDLINE | ID: mdl-34708965

RESUMO

Dissolved organic matter(DOM) in water environments is an important component of the global carbon cycle. Under the current urgency to control the pollution of urban rivers in China, exploring the influence of different exogenous and endogenous secondary pollution and weather patterns on river DOM is the premise to better understand the causes of the pollution. In this study, a large city in China was established as the research area, and the underlying water and sediments from 21 sites along urban and suburban rivers, and other water sources were evaluated. The excitation-emission matrix-parallel factor analysis(EEM-PARAFAC) was used to analyze the difference in DOM composition and equivalent in urban rivers polluted by domestic sewage and suburban rivers polluted by aquaculture, agriculture, and livestock breeding. The results showed that:① DOM components in urban and suburban river waters were mainly protein compounds(tyrosine-and tryptophan-like), containing a small amount of humic acid. Humic acid components of anthropogenic origin were found in urban river water; ② The reasons for the formation of DOM components in urban and suburban rivers were completely different. Urban rivers are mainly polluted by domestic sewage and endogenous secondary pollution, resulting in an increase in anthropogenic humic acid components. Suburban rivers are mainly polluted by agricultural wastewater rich in N and P, which promotes endogenous metabolism of autotrophic bacteria and increases protein components, which may be related to the formation of existing DOM characteristics. ③ Rainfall runoff and urban overflow transported exogenous pollutants into rivers, while hydrodynamic factors such as hydraulic agitation affect the distribution of DOM components in underlying water and sediments through physical effects such as dilution.


Assuntos
2,5-Dimetoxi-4-Metilanfetamina , Rios , Análise Fatorial , Substâncias Húmicas/análise , Espectrometria de Fluorescência , Águas Residuárias
9.
J Glob Antimicrob Resist ; 27: 253-266, 2021 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-34700054

RESUMO

OBJECTIVES: Candida albicans is the most clinically prevalent cause of systemic fungal infections in the immunocompromised population. The biofilm-forming ability of C. albicans confers resistance to conventional antifungal agents. The main aim of this study was to investigate the antifungal effects of ethyl caffeate (EC) alone and in combination with fluconazole (FLU) against C. albicans isolates. METHODS: The single and combined antifungal activities of EC and FLU were evaluated against planktonic and biofilm cells of C. albicans by the checkerboard assay, time-kill test, crystal violet assay, live/dead staining, rhodamine 6G (R6G) efflux analysis and hydrolase activity. Monotherapy and dual therapy of EC and FLU against systemic candidiasis in a mouse model was also evaluated. RESULTS: The results showed that EC+FLU displayed synergism in 14/26 planktonic C. albicans isolates and 11/26 C. albicans biofilms with fractional inhibitory concentration index (FICI) values ranging between 0.06-0.49 and 0.02-0.38, respectively. Compared with monotherapy, the combination of EC+FLU can markedly inhibit adhesion, yeast-to-hyphae transition, premature and mature biofilm metabolism, hydrolase secretion and drug efflux function of C. albicans Z1407 and Z4935. Moreover, EC can potentiate the antifungal activity of FLU to improve mouse survival, reduce fungal burden and alleviate pathological damage in both C. albicans isolates compared with EC or FLU used alone. CONCLUSION: EC exhibits a moderate antifungal potential but can be a strong synergist with FLU against C. albicans, highlighting the potential of EC in clinical antifungal therapy as a sensitiser.

10.
Front Surg ; 8: 726067, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34568419

RESUMO

Objective: The present study aimed to explore the application value of magnetic resonance imaging (MRI) histograms with multiple sequences in the preoperative differential diagnosis of endometrial stromal sarcoma (ESS) and degenerative hysteromyoma (DH). Methods: The clinical and preoperative MRI data of 20 patients with pathologically confirmed ESS and 24 patients with pathologically confirmed DH were retrospectively analyzed, forming the two study groups. Mazda software was used to select the MRI layer with the largest tumor diameter in T2WI, the apparent diffusion coefficient (ADC), and enhanced T1WI (T1CE) images. The region of interest (ROI) was outlined for gray-scale histogram analysis. Nine parameters-the mean, variance, kurtosis, skewness, 1st percentile, 10th percentile, 50th percentile, 90th percentile, and 99th percentile-were obtained for intergroup analysis, and the receiver operating curves (ROCs) were plotted to analyze the differential diagnostic efficacy for each parameter. Results: In the T2WI histogram, the differences between the two groups in seven of the parameters (mean, skewness, 1st percentile, 10th percentile, 50th percentile, 90th percentile, and 99th percentile) were statistically significant (P < 0.05). In the ADC histogram, the differences between the two groups in three of the parameters (skewness, 10th percentile, and 50th percentile) were statistically significant (P < 0.05). In the T1CE histogram, no significant differences were found between the two groups in any of the parameters (all P > 0.05). Of the nine parameters, the 50th percentile was found to have the best diagnostic efficacy. In the T2WI histogram, ROC curve analysis of the 50th percentile yielded the best area under the ROC curve (AUC; 0.742), sensitivity of 70%, and specificity of 83.3%. In the ADC histogram, ROC curve analysis of the 50th percentile yielded the best area under the ROC curve (AUC; 0.783), sensitivity of 81%, and specificity of 76.9%. Conclusion: The parameters of the mean, 10th percentile and 50th percentile in the T2WI histogram have good diagnostic efficacy, providing new methods and ideas for clinical diagnosis.

11.
J Med Chem ; 64(19): 14895-14911, 2021 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-34546748

RESUMO

The major drawbacks of P-glycoprotein (P-gp) inhibitors at the clinical stage make the development of new P-gp inhibitors challenging and desirable. In this study, we reported our structure-activity relationship studies of 4-indolyl quinazoline, which led to the discovery of a highly effective and orally active P-gp inhibitor, YS-370. YS-370 effectively reversed multidrug resistance (MDR) to paclitaxel and colchicine in SW620/AD300 and HEK293T-ABCB1 cells. YS-370 bound directly to P-gp, did not alter expression or subcellular localization of P-gp in SW620/AD300 cells, but increased the intracellular accumulation of paclitaxel. Furthermore, YS-370 stimulated the P-gp ATPase activity and had moderate inhibition against CYP3A4. Significantly, oral administration of YS-370 in combination with paclitaxel achieved much stronger antitumor activity in a xenograft model bearing SW620/Ad300 cells than either drug alone. Taken together, our data demonstrate that YS-370 is a promising P-gp inhibitor capable of overcoming MDR and represents a unique scaffold for the development of new P-gp inhibitors.

12.
Orthop Surg ; 13(7): 2000-2007, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34523793

RESUMO

OBJECTIVE: To analyze correlations between the selection of microtitanium plates with different specifications for use in a cervical vertebral dome expansion laminoplasty. METHODS: Sixteen patients that underwent the cervical vertebral dome expansion laminoplasty with a cervical spinal stenosis angioplasty procedure for treatment of their cervical spinal cords were recruited at our hospital. From February 2017 to September 2018, medical records confirmed that all patients underwent cervical CT and MRI tests pre- and postsurgery. The anteroposterior diameter of the spinal canal, changes in the cross-sectional area of the spinal canal, and the pre- and postsurgery distance of the cervical spinal cord after applying microtitanium plates with different lengths were measured by Mimics version 17.0 software (Materialise NV, Leuven, Belgium). A statistical regression and correlation analysis of relevant specification parameters of the microtitanium plate was then studied. RESULTS: As the size of the microtitanium plate increased, we found that the cross-sectional area of cervical spinal canal and distance between the descendants of the lamina and the distance of cervical spinal cord concordantly increased, and these data changes linearly. The regression equation associated with sagittal diameter, cross-sectional area, and posterior movement distance of the cervical spinal cord was obtained. CONCLUSION: According to the correlation analysis of imaging data changes, the regression equation was obtained to guide the selection of microtitanium plates with appropriate specifications in a cervical vertebral dome expansion laminoplasty.


Assuntos
Placas Ósseas , Vértebras Cervicais/cirurgia , Desenho de Equipamento , Laminoplastia/instrumentação , Estenose Espinal/cirurgia , Idoso , Vértebras Cervicais/diagnóstico por imagem , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estenose Espinal/diagnóstico por imagem , Titânio , Tomografia Computadorizada por Raios X
13.
Orthop Surg ; 13(7): 1969-1978, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34523808

RESUMO

OBJECTIVE: To evaluate the safety and stability of 3D-printed interbody fusion cages (3D-printed cages) in anterior cervical discectomy and fusion (ACDF) by investigating the mid- and long-term follow-up outcomes. METHODS: In this prospective study, the clinical data of 30 patients with CSM admitted to the Second Hospital of Shanxi Medical University from May 2012 to May 2014 were analyzed. The cohort comprised 18 males and 12 females with an average age of 60.22 ± 3.2 years. All patients were examined by X-ray, CT and MRI before the operation. A total of 30 cases of CSM were treated by ACDF with 3D printed cage implantation. Mid- and long-term follow-ups were performed after the surgery. Clinical efficacy was evaluated by comparing the JOA score, SF-36 score, change in neurological function, cervical curvature index (CCI), vertebral intervertebral height (VIH) and fusion rate before the operation, 6 months after the operation, and at the last follow-up. RESULTS: Two of the 30 patients were lost to follow-up. The remaining patients were followed up for 48-76 (65.23 ± 3.54) months. The patients recovered satisfactorily with a significant clinical effect. The JOA score increased meanfully and the improvement rate was 89.4% at the final follow-up. The SF-36 score increased significantly from pre- to postoperatively. The height of the intervertebral space at the last follow-up was not statistically significantly different from that at 6 months after surgery (P > 0.05), showing that the height of the intervertebral space did not change much and the severity of cage subsidence (CS) decreased. The CCI improved from pre- to postoperatively. The CCI did not change much from the 6-month follow-up to the last follow-up. and the cage rate (CR) was 100% at the 6-month and last follow-ups. No severe complications, such as spinal cord injury, esophageal fistula, cerebrospinal fluid leakage, cervical hematoma or wound infection, occurred in any of the patients. CONCLUSION: The clinical and radiological results show that the application of 3D-printed cages in ACDF can significantly relieve symptoms. Moreover, 3D-printed cages can restore the curvature of the cervical spine, effectively maintain the intervertebral height for a long time, and prevent complications related to postoperative subsidence.


Assuntos
Vértebras Cervicais/cirurgia , Discotomia/métodos , Impressão Tridimensional , Doenças da Medula Espinal/cirurgia , Fusão Vertebral/instrumentação , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do Tratamento
14.
Chem Commun (Camb) ; 57(76): 9748-9751, 2021 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-34477187

RESUMO

Guanosine deaminase (GSDA) in plants specifically deaminates (de)guanosine to produce xanthosine with high specificity, which is further converted to xanthine, a key intermediate in purine metabolism and nitrogen recycling. We solved GSDA's structures from Arabidopsis thaliana in the free and ligand-bound forms at high resolutions. Unlike GDA, the enzyme employs a single-proton shuttle mechanism for catalysis and both the substrate and enzyme undergo structural rearrangements. The last fragment of the enzyme loops back and seals the active site, and the substrate rotates during the reaction, both essential to deamination. We further identified more substrates that could be employed by the enzyme and compare it with other deaminases to reveal the recognition differences of specific substrates. Our studies provide insight into this important enzyme involved in purine metabolism and will potentially aid in the development of deaminase-based gene-editing tools.


Assuntos
Arabidopsis/enzimologia , Nucleosídeo Desaminases/metabolismo , Biocatálise , Modelos Moleculares , Nucleosídeo Desaminases/química
15.
Front Plant Sci ; 12: 687954, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34335655

RESUMO

Nitrogen (N) and sulfur (S) are essential mineral nutrients for plant growth and metabolism. Here, we investigated their interaction in plant growth and andrographolide accumulation in medicinal plant Andrographis paniculata grown at different N (4 and 8 mmol·L-1) and S concentration levels (0.1 and 2.4 mmol L-1). We found that increasing the S application rate enhanced the accumulation of andrographolide compounds (AGCs) in A. paniculata. Simultaneously, salicylic acid (SA) and gibberellic acid 4 (GA4) concentrations were increased but trehalose/trehalose 6-phosphate (Tre/Tre6P) concentrations were decreased by high S, suggesting that they were involved in the S-mediated accumulation of AGCs. However, S affected plant growth differentially at different N levels. Metabolite analysis revealed that high S induced increases in the tricarboxylic acid (TCA) cycle and photorespiration under low N conditions, which promoted N assimilation and S metabolism, and simultaneously increased carbohydrate consumption and inhibited plant growth. In contrast, high S reduced N and S concentrations in plants and promoted plant growth under high N conditions. Taken together, the results indicated that increasing the S application rate is an effective strategy to improve AGC accumulation in A. paniculata. Nevertheless, the interaction of N and S affected the trade-off between plant growth and AGC accumulation, in which N metabolism plays a key role.

16.
Exp Ther Med ; 22(3): 938, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34335887

RESUMO

The aim of the current study was to develop an accurate and reproducible method for isolation of granulosa cells (GCs) in patients with different ovarian reserves. The cells of healthy individuals and patients with polycystic ovary syndrome (PCOS) were isolated using a modified two-step Percoll density gradient centrifugation. The cells of patients with poor ovarian response (POR) were isolated using a one-step method suitable for samples containing few cells. Cells extracted using these purification techniques were compared regarding cell yield, viability and purity using immunocytochemistry, flow cytometry, Cell Counting Kit-8, western blotting and RNA integrity analysis. The purity and activity of the cells in the POR group were comparable with those in the healthy and PCOS groups and no statistically significant differences were identified. Furthermore, isolated cells analyzed for RNA integrity indicated good quality RNA and presented an RNA integrity number of 8-10, demonstrating that the technique enabled the isolation of GCs from different types of patients. Thus, a reliable and reproducible technique for the isolation of pure GCs with high yield is described in the present study. This protocol provides an efficient method targeted to patients with different ovarian reserve functions that enables the preparation of GCs for evaluating their molecular functions.

17.
Insects ; 12(7)2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-34357261

RESUMO

Spodoptera litura F. is a generalist herbivore and one of the most important economic pests feeding on about 300 host plants in many Asian countries. Specific insect behaviors can be stimulated after recognizing chemicals in the external environment through conserved chemosensory proteins (CSPs) in chemoreceptive organs, which are critical components of the olfactory systems. To explore its structural basis for ligand-recognizing capability, we solved the 2.3 Å crystal structure of the apoprotein of S. litura CSP8 (SlCSP8). The SlCSP8 protein displays a conserved spherical shape with a negatively charged surface. Our binding assays showed that SlCSP8 bound several candidate ligands with differential affinities, with rhodojaponin III being the most tightly bound ligand. Our crystallographic and biochemical studies provide important insight into the molecular recognition mechanism of the sensory protein SlCSP8 and the CSP family in general, and they suggest that CSP8 is critical for insects to identify rhodojaponin III, which may aid in the CSP-based rational drug design in the future.

18.
Nat Metab ; 3(8): 1109-1124, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34385701

RESUMO

Zika virus (ZIKV) infection during pregnancy can cause microcephaly in newborns, yet the underlying mechanisms remain largely unexplored. Here, we reveal extensive and large-scale metabolic reprogramming events in ZIKV-infected mouse brains by performing a multi-omics study comprising transcriptomics, proteomics, phosphoproteomics and metabolomics approaches. Our proteomics and metabolomics analyses uncover dramatic alteration of nicotinamide adenine dinucleotide (NAD+)-related metabolic pathways, including oxidative phosphorylation, TCA cycle and tryptophan metabolism. Phosphoproteomics analysis indicates that MAPK and cyclic GMP-protein kinase G signaling may be associated with ZIKV-induced microcephaly. Notably, we demonstrate the utility of our rich multi-omics datasets with follow-up in vivo experiments, which confirm that boosting NAD+ by NAD+ or nicotinamide riboside supplementation alleviates cell death and increases cortex thickness in ZIKV-infected mouse brains. Nicotinamide riboside supplementation increases the brain and body weight as well as improves the survival in ZIKV-infected mice. Our study provides a comprehensive resource of biological data to support future investigations of ZIKV-induced microcephaly and demonstrates that metabolic alterations can be potentially exploited for developing therapeutic strategies.


Assuntos
Microcefalia/etiologia , Microcefalia/metabolismo , NAD/metabolismo , Infecção por Zika virus/complicações , Infecção por Zika virus/virologia , Zika virus/fisiologia , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Encéfalo/virologia , Células Cultivadas , Cromatografia Líquida , Modelos Animais de Doenças , Suscetibilidade a Doenças , Feminino , Metabolômica , Camundongos , Microcefalia/patologia , Neurônios/metabolismo , Gravidez , Proteômica/métodos , Espectrometria de Massas em Tandem
19.
Artigo em Inglês | MEDLINE | ID: mdl-34461303

RESUMO

BACKGROUND & AIMS: Both environmental factors, such as alcohol consumption and smoking, and genetic factors are strongly associated with the risk of developing chronic pancreatitis (CP). However, comprehensive understanding of their impacts on the progression of CP remains elusive. METHODS: A prospective cohort study was performed on a large cohort of CP patients with known genetic backgrounds. The cumulative incidence of pancreatic insufficiency after the onset of CP was analyzed using Kaplan-Meier survival curves. Multivariate Cox proportional hazards regression analysis also was performed. RESULTS: A total of 798 patients were enrolled in the study and followed up for 10.5 years. Rare pathogenic genotypes in the SPINK1, PRSS1, CTRC, or CFTR genes were identified in 410 (51.4%) patients. The development of pancreatic insufficiency was significantly earlier in patients with a history of smoking and/or alcohol consumption in both the positive (P < .001) and negative (P = .001) gene mutation groups. However, the development of pancreatic insufficiency did not differ significantly between patients with and without gene mutations despite alcohol and/or smoking status, with P values of .064 and .115, respectively. Multivariate Cox regression analysis showed that age at onset of CP (hazard ratio, [HR], 1.02; P < .001) and alcohol consumption (HR, 1.86; P < .001) were independent risk factors for the development of diabetes, while male sex (HR, 1.84; P = .022) and smoking (HR, 1.56; P = .028) were predictors of steatorrhea. CONCLUSIONS: Although rare pathogenic mutations in the 4 major susceptibility genes for CP were not correlated significantly with the development of pancreatic insufficiency, environmental factors (either alcohol consumption or smoking) significantly accelerated disease progression (ClinicalTrials.gov: NCT04574297).

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