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1.
Chem Commun (Camb) ; 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31690902

RESUMO

We developed a novel fluorescence labelling method to track exosome internalization pathways in cells by confocal microscopy. The proposed method allows evaluation and comparison of the uptake pathways of exosomes originating from different cells, which would offer the potential for understanding the functions of exosomes in intercellular communication and their applications in drug delivery.

2.
Molecules ; 24(21)2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31694145

RESUMO

Carbon fiber mesh reinforced cement-based composites (CMCCs) have received extensive attention in the field of engineering repair and structural reinforcement due to their outstanding properties such as two-way force, rust prevention, high specific strength, and low base surface requirements. However, the development of this material has been slowed down to some extent due to the poor interfacial bonding between the carbon fiber mesh and the cement matrix. In this paper, a novel fabrication strategy was proposed in which the carbon fiber mesh was modified with epoxy resin and silane coupling agent (SCA) to increase its surface chemical activity. Meanwhile, the hydroxymethyl cellulose (HMC) was also filled into the concrete matrix to improve the mechanical strength of the matrix as well as the load transfer behaviors between the mortar and carbon fiber (CF) mesh. The potential to employ SCA and HMC was evaluated for the making of CMCCs via the above methods. The results showed that the longitudinal shear strength of composites with SCA and SCA&HMC increased by 26.6% and 56.1% compared to those of CF with epoxy resin (EP) reinforced composites, respectively. The flexural strength of composite with SCA&HMC increases by 147.6% compared to I-(F) without CF. The novel II-HCM&CF/EP-SCA composites with excellent performance are promised to be applied in practical uses.

3.
PLoS One ; 14(11): e0224876, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31703095

RESUMO

Gene-environment interactions contribute to the risk for Autism Spectrum Disorder (ASD). Among environmental factors, prenatal exposure to stress may increase the risk for ASD. To examine if there is an interaction between exposure to maternal stress and reduced dosage or loss of Shank3, wild-type (WT), heterozygous (HET) and homozygous (HOM) female mice carrying a deletion of exons four through nine of Shank3 (Shank3ex4-9) were exposed to chronic unpredictable mild stress (CUMS) from prior to conception throughout gestation. This study examined maternal care of these dams and the white matter microstructure in the brains of their adult male offspring. Overall, our findings suggest that maternal exposure to CUMS increased pup-directed care for dams of all three genotypes. Compared to WT and HET dams, HOM dams also exhibited increased maternal care behaviors with increased time spent in the nest and reduced cage exploration, regardless of exposure to CUMS. Diffusion tensor imaging showed higher mean fractional anisotropy in the hippocampal stratum radiatum of WT and HOM male offspring from dams exposed to CUMS and HOM offspring from unexposed dams, compared to WT male offspring from unexposed dams. These data support that CUMS in Shank3-mutant dams results in subtle maternal care alterations and long-lasting changes in the white matter of the hippocampus of their offspring.

4.
J Cell Physiol ; 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31674022

RESUMO

Gastric cancer continues to be a common cancer in the world with high incidence and mortality. Accumulating evidence has implicated long noncoding RNAs (lncRNAs) in gastric cancer progression. Here, this study identified the potential role of a novel lncRNA, LINC00629 in gastric cancer and to elucidate the underlying mechanism. Initially, microarray-based gene expression profiling of gastric cancer was employed to identify differentially expressed genes. Next, the expression of LINC00629, microRNA-196b-5p (miR-196b-5p) and aquaporin 4 (AQP4) in clinical gastric cancer tissues was determined and the cell line presenting with the lowest LINC00629 expression was selected. The interaction among LINC00629, miR-196b-5p, and AQP4 was identified. Expression of LINC00629, miR-196b-5p, and AQP4 in gastric cancer cells were altered and then biological behaviors of gastric cancer cells were assessed by 5-ethynyl-2'-deoxyuridine and Transwell assays. Tumor formation in vivo was evaluated in nude mice. In gastric cancer, expression of LINC00629 and AQP4 was downregulated, and expression of miR-196b-5p was upregulated. Proliferation, invasion, and migration of gastric cancer cells were reduced after overexpression of LINC00629. LINC00629 competitively bound to miR-196b-5p, while AQP4 was a target of miR-196b-5p. Either downregulating miR-196b-5p or upregulating AQP4 could restrain the development of gastric cancer in vitro. LINC00629 overexpression repressed the growth of transplanted tumors in vivo. Taken together, LINC00629 competitively bound to miR-196b-5p to upregulate AQP4 expression, thereby inhibiting gastric cancer progression. Therefore, understanding of this mechanism may help to improve gastric cancer treatment.

5.
Artigo em Inglês | MEDLINE | ID: mdl-31698108

RESUMO

OBJECTIVES: The acquisition of carbapenemase is of particular concern for Proteus mirabilis, which is intrinsically resistant to tigecycline and colistin, as this will make clinical therapy extremely difficult. Here we report the whole genome sequence of one P. mirabilis clinical isolate harboring blaNDM-5 on an IncX3 type plasmid from China. METHODS: Whole genome sequencing of the isolate was sequenced using Illumina HiseqTM 4000 and MinION. Hybrid assembly of both short Illumina reads and long MinION reads was performed using Unicycler v 0.4.7. Functional annotation was performed by the NCBI Prokaryotic Genomes Annotation Pipeline (PGAP) server. Genomic analysis was further performed. RESULTS: The complete genome sequence of P. mirabilis CRPM10 was consisted of one chromosomal DNA comprising 4,158,695 bp and one plasmid with size of 46,161 bp. Fourteen resistance genes were identified in CRPM10. Resistance genes were all located on the chromosomal DNA except for blaNDM-5, which was located on a 46,161 bp IncX3 type plasmid named pNDM-5. Plasmid sequence alignment of pNDM-5 with the NCBI GenBank database revealed several plasmids from different Enterobacteriaceae strains are highly identical. CONCLUSIONS: We report the complete genome sequence of a P. mirabilis clinical isolate carrying blaNDM-5 on an IncX3 type plasmid from China. The 46,161 bp IncX3-type plasmids play an important role in the distribution of NDM mutants among Enterobacteriaceae strains. Considering the global emergence of carbapenemase NDM-5, an epidemiological survey and analysis of blaNDM-5 harbouring Enterobacteriaceae strains are urgently needed to prevent its future prevalence.

6.
Infect Immun ; 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31570555

RESUMO

Pasteurella multocida causes a variety of infectious diseases in various species of mammals and birds, resulting in enormous economic loss to the modern livestock and poultry industry. However, the mechanism of host-pathogen interactions is unclear. Here we found that L-serine levels were significantly decreased in murine lungs infected with P. multocida Exogenous L-serine supplementation significantly increased the survival rate of mice, and decreased the colonization of P. multocida in the lungs of mice. Notably, L-serine decreased the macrophage- and neutrophil-mediated inflammatory responses in mice during P. multocida infection.

7.
Artigo em Inglês | MEDLINE | ID: mdl-31600810

RESUMO

OBJECTIVE: The clinical data of patients with hemifacial spasm (HFS) were analyzed statistically to identify factors leading to delayed cure after microvascular decompression (MVD). METHODS: A retrospective analysis of the clinical data of 600 patients with HFS subjected to MVD from March 2016 to May 2018 was performed. Student t test, chi-square test, logistic regression analysis, and multivariate analysis of variance were used to analyze the correlation between delayed cure and its related factors. RESULTS: Among the 600 patients enrolled, 117 had delayed cure after MVD. The shortest duration of delayed cure was 4 days, and the longest was 540 days, with an average of 108 days. The frequency of delayed improvement in these patients was not associated with sex, age, or offending vessel type (p > 0.05); however, delayed cure was positively correlated with the course of the disease, grade of HFS severity, and disappearance of abnormal muscle responses during the operation (p < 0.05). Moreover, a longer disease course was associated with more severe related symptoms and a longer duration of postoperative delayed cure. CONCLUSION: MVD is an effective treatment for HFS. Given that postoperative delayed cure was unavoidable, even with accurate identification of the offending vessel and sufficient decompression of the root exit zone, delayed cure should be considered in patients undergoing reoperation due to lack of remission or relapse after the operation. Additionally, the timing of efficacy assessments should be delayed.

8.
Wei Sheng Yan Jiu ; 48(4): 594-600, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31601341

RESUMO

OBJECTIVE: To understand the prevalence and influencing factors of food hypersensitivity in 1-year-old infants in Chengdu. METHODS: A two-way cohort study was conducted to select 923 pairs of mothers and infants from the Second Hospital of Sichuan University from March 2014 to March 2015. The questionnaires and telephone follow-up surveys were used to investigate the incidence and influencing factors of parents' self-reported food hypersensitivity of infants. RESULTS: The prevalence of parents' self-reported food hypersensitivity was 14. 1% before the age of one year, and the average age of the first onset was(6. 9±3. 3) month old. The main manifestations were skin(85. 7%) and gastrointestinal symptoms(14. 3%), those were mainly caused by aquatic products(42. 3%), eggs(26. 0%), and milk(22. 1%). Family history of allergy, consumption of milk during pregnancy, exposure to pets during pregnancy, and addition of seafood within 6 months of age are risk factors for parents' self-reported food hypersensitivity in infants. Adding egg protein as early as possible within 1 year old may reduce the incidence of parents' self-reported food hypersensitivity of infants. CONCLUSION: Preventing food hypersensitivity should start from the beginning of pregnancy. Different types of complementary food have different effects on the occurrence of food hypersensitivity in infants. Personalized management of complementary food adding should be conducted according to infant genetic history, living environment and epidemiological survey data to preventing food hypersensitivity.

9.
Cancer Control ; 26(1): 1073274819883895, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31642331

RESUMO

Identifying metastasis remains a challenge for death control and tailored therapy for nasopharyngeal carcinoma (NPC). Here, we addressed this by designing a nomogram-based Cox proportional regression model through integrating a panel of tumor biomarkers. A total of 147 locally patients with advanced NPC, derived from a randomized phase III clinical trial, were enrolled. We constructed the model by selecting the variables from 31 tumor biomarkers, including 6 pathological signaling pathway molecules and 3 Epstein-Barr virus-related serological variables. Through the least absolute shrinkage and selection operator (LASSO) Cox proportional regression analysis, a nomogram was designed to refine the metastasis risk of each NPC individuals. Using the LASSO Cox regression model, we constructed a 9 biomarkers-based prognostic nomogram: Beclin 1, Aurora-A, Cyclin D1, Ki-67, P27, Bcl-2, MMP-9, 14-3-3σ, and VCA-IgA. The time-dependence receiver operating characteristic analysis at 1, 3, and 5 years showed an appealing prognostic accuracy with the area under the curve of 0.830, 0.827, and 0.817, respectively. In the validation subset, the concordance index of this nomogram reached to 0.64 to identify the individual metastasis pattern. Supporting by this nomogram algorithm, the individual metastasis risk might be refined personally and potentially guiding the treatment decisions and target therapy against the related signaling pathways for patients with locally advanced NPC.

10.
PLoS One ; 14(10): e0223665, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31618237

RESUMO

OBJECTIVE: We aim to explore the relationship between nocturnal sleep duration (NSD) and midday nap duration (MND) with body composition among Southwest Chinese adults. METHODS: Data on sleep duration of 3145 adults in Southwest China (59.4% women) were obtained between 2014 and 2015 through questionnaires. Height, weight, and waist circumference (WC) were measured to calculate body composition (body mass index (BMI), percentage of body fat (%BF), and fat mass index (FMI)). Linear regression models were used to assess gender-specific associations between NSD and body composition. The relationship between MND with the odds of overweight and central obesity has been evaluated by logistic regression models. RESULTS: NSD has the inverse relation with males' BMI, WC, %BF and FMI after adjusting for all covariates (all P <0.0007), exclusive of females' (all P >0.4). After adjustment for potential confounders, compared to the subjects in the no midday nap group, the subjects who napped 0.1-1 hour were independently associated with a less prevalence of overweight in both women (OR: 0.72, 95%CI: 0.55-0.95) and men (OR: 0.71, 95%CI: 0.52-0.98). MND was not associated with central obesity. CONCLUSIONS: Among Southwest Chinese adults, lower NSD might be related to higher BMI, WC, %BF and FMI among men. Additionally, MND is associated with overweight in adults.

11.
Biosci Biotechnol Biochem ; : 1-10, 2019 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-31661358

RESUMO

This study assessed whether antibiotics could alter gut microbiota to affect host growth and the possibility of alleviation by lactobacilli. We divided four-week-old BABL/c mice into control (Ctrl), antibiotic exposure (Abx), Lactobacillus plantarum PC-170 (PC), and Lactobacillus rhamnosus GG (LGG) group and the Abx, LGG, and PC group received an one-week antibiotic/antibiotic + probiotic treatment. The fecal microbiota and the expression of splenic cytokines were determined. Following the ceftriaxone treatment, the body weight gain of Abx was delayed compared with others. The ceftriaxone treatment significantly decreased the alpha-diversity of the fecal microbiota and altered the fecal microbiota but LGG and PC can partly alleviate the effect. At the end of the study, the microbial community of LGG and PC group were more similar to Ctrl compared with Abx group. The results indicated that ceftriaxone could significantly alter intestinal microbiota. Lactobacilli might alleviate the side effects of antibiotics by stabilizing the intestinal microbiota.

12.
Gut ; 2019 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-31563875

RESUMO

OBJECTIVE: Fibrosis stage is strongly associated with liver-related outcomes and is a key surrogate endpoint in drug trials for non-alcoholic steatohepatitis. Dual-photon microscopy allows automated quantification of fibrosis-related parameters (q-FPs) and may facilitate large-scale histological studies. We aim to validate the performance of q-FPs in a large histological cohort. DESIGN: 344 patients with non-alcoholic fatty liver disease (NAFLD) underwent 428 liver biopsies (240 had paired transient elastography examination). Fibrosis stage was scored using the NASH Clinical Research Network system, and q-FPs were measured by dual-photon microscopy using unstained slides. Patients were randomly assigned to the training and validation cohorts to test the performance of individual q-FPs and derive optimal cut-offs. RESULTS: Over 25 q-FPs had area under the receiver-operating characteristics curves >0.90 for different fibrosis stages. Among them, the perimeter of collagen fibres and number of long collagen fibres had the highest accuracy. At the best cut-offs, the two q-FPs had 88.3%-96.2% sensitivity and 78.1%-91.1% specificity for different fibrosis stages in the validation cohort. q-FPs and histological scoring had nearly identical correlations with liver stiffness measurement, suggesting that the accuracy of q-FPs approached that of histological assessment. Among patients with paired liver biopsies, changes in the same q-FPs were associated with changes in fibrosis stage. At a median follow-up of 5.6 years, baseline q-FPs predicted liver-related events. CONCLUSION: q-FP is highly accurate in the assessment of fibrosis in NAFLD patients. This automated platform can be used in future studies as objective and reliable evaluation of histological fibrosis.

13.
J Matern Fetal Neonatal Med ; : 1-8, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31530067

RESUMO

Objective: Current evidence for negative pressure wound therapy (NPWT) on surgical site infection (SSI) and wound complications in cesarean section is conflicting. The objective of this study was to evaluate the efficacy of prophylactic NPWT for preventing SSI and other wound complications in obese women undergoing cesarean section (CS). Methods: We systematically searched PubMed, Embase, the Cochrane Library and clinicaltTrial.gov to identify randomized controlled trials (RCTs) that compared NPWT with standard dressing for cesarean section. The primary outcome was SSI. Secondary outcomes were overall wound complications and hospital readmission. Risk ratio (RR) with 95% confidence intervals (CIs) was calculated using random-effects models. Review Manager 5.3 was applied to analyze the collected data. Results: Eight RCTs involving 1972 patients were included in this meta-analysis. The pooled results showed that the risk of SSI was significantly lower with the use of NPWT when compared with standard dressing (RR = 0.68, 95%CI = 0.51-0.90, p = .008). However, there was no difference in overall wound complications (RR = 0.93, 95%CI = 0.74-1.17, p = 0.52) and hospital readmission (RR = 1.03, 95%CI = 0.67-1.60, p = .89) between two groups. Current evidence was not confirmed by trial sequential analysis. Conclusion: On the basis of our findings, NPWT decreases the risk of SSI after cesarean section in obese women after CS, despite this approach does not reduce the overall wound complications and hospital readmission. However, further RCTs are needed to make conclusive evidence.

14.
Mol Med Rep ; 20(5): 4433-4448, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31485595

RESUMO

Hypertension is a cardiovascular disease that severely impairs human health; however, its specific etiology and pathogenesis are complex. The present study investigated the effects of the calcium sensing receptor (CaSR) on vascular tone in spontaneously hypertensive rats (SHRs), and clarified the role and mechanism of CaSR in regulating this property with respect to the phospholipase C (PLC)­inositol 1,4,5­triphosphate (IP3)/adenylate cyclase­V(AC­V)/cyclic adenosine monophosphate (cAMP)/renin­angiotensin system (RAS) pathway in these animals. CaSR protein expression in the mesenteric artery (MA) of rats and CaSR protein expression in SHRs were significantly reduced. Based on wire myography studies, vasoconstriction was significantly augmented and vasodilatation was attenuated in SHRs, and this effect was endothelium­independent. The CaSR calcimimetic NPSR568 and inhibitor NPS2143 reduced vasoconstriction and enhanced vasodilation in SHRs. Furthermore, pretreatment with PLC­IP3/AC­V/cAMP/RAS pathway blockers significantly reduced the vasoconstriction response and enhanced the vasodilator response in SHRs and Wistar­Kyoto rats (WKY), and these effects were partially dependent on the endothelium. Additionally, pretreatment with CaSR inhibitors were determined to cooperate with the PLC­IP3/AC­V/cAMP/RAS pathway inhibitors to significantly reduce vasoconstriction and enhance vasodilation in SHRs and WKY. Our results demonstrated that CaSR is functionally expressed in the MA of SHRs, and that CaSR expression is decreased in SHRs. Additionally, vasoconstriction was enhanced while vasodilatation was attenuated in SHRs; these processes were determined to be endothelium­independent. CaSR is involved in the regulation of blood pressure and vascular tension in SHRs and WKYs. In association with mechanistic differences, this effect was proposed to be partially endothelium­dependent and mediated by the PLC­IP3/AC­V/cAMP/RAS pathway.

15.
Mol Med Rep ; 20(5): 4277-4284, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31545409

RESUMO

The objective of the present study was to investigate the effects of polo­like kinase 1 (PLK1) and the phosphorylation of human cell division cycle protein 14A (Cdc14A) by PLK1 on ß­cell function and cell cycle regulation. Mouse ß­TC3 cells were incubated with small interfering RNA (siRNA) to knock down the expression of PLK1. Cell cycle analysis was performed using flow cytometry, and cell proliferation and apoptosis was determined. Insulin secretion was evaluated by a radioimmunoassay under both low and high glucose conditions. Mouse ß­TC3 cells were transfected with a wild type or a non­phosphorylatable Cdc14A mutant (Cdc14AS351A/363A; Cdc14AAA) to investigate whether the phosphorylation of Cdc14A is involved in cellular regulation of PLK1 under high glucose conditions. It was found that PLK1 siRNA significantly promoted cellular apoptosis, inhibited cell proliferation, decreased insulin secretion and reduced Cdc14A expression under both low and high glucose conditions. Cdc14A overexpression promoted ß­TC3 cell proliferation and insulin secretion, while Cdc14AAA overexpression inhibited cell proliferation and insulin secretion under high glucose conditions. PLK1 siRNA partially reversed the proliferation­promoting effects of Cdc14A and further intensified the inhibition of proliferation by Cdc14AAA under high glucose conditions. Similarly, Cdc14A overexpression partially reversed the insulin­inhibiting effects of PLK1 siRNA, while Cdc14AAA overexpression showed a synergistic inhibitory effect on insulin secretion with PLK1 siRNA under high glucose conditions. In conclusion, PLK1 promoted cell proliferation and insulin secretion while inhibiting cellular apoptosis in ß­TC3 cell lines under both low and high glucose conditions. In addition, the phospho­regulation of Cdc14A by PLK1 may be involved in ß­TC3 cell cycle regulation and insulin secretion under high glucose conditions.

16.
Biomed Pharmacother ; 118: 109279, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31376651

RESUMO

COX-2 specific inhibitor, which has been widely used, can delay bone fracture healing and reduce osteogenic potential of bone marrow stromal cells. However, it remains unknown how to prevent these side-effects of COX-2 inhibitor. In this study, we introduced BMP9-induced osteogenic differentiation as model to evaluate whether all-trans retinoic acid (ATRA) could ameliorate these adverse effects of COX-2 specific inhibitor on bone metabolism with in vitro and in vivo experiments, and uncover the possible mechanism underlying this process. Results showed that ATRA enhanced the potential of BMP9 to induce the osteogenic markers, such as alkaline phosphates (ALP) and mineralization; but retinoic acid receptor a (RARa) inhibitor showed the reversal effects. COX-2 specific inhibitor (NS398) reduced the osteogenic markers induced by BMP9, and ATRA almost eliminated the inhibitory effect of NS398. BMP9 up-regulated the protein level of ß-catenin and promoted it translocate to nucleus, and both were reduced by NS398. On the contrary, ATRA notablely attenuated the inhibitory effect of NS398 on BMP9-increased ß-catenin. Exogenous RXRa obviously ameliorated the inhibitory effect of silencing COX-2 on ectopic bone formation induced by BMP9. NS398 reduced the level of phosphorylated CREB, which was almost reversed by ATRA. Besides, RXRa interacted with phosphorylated CREB directly and both were recruited at ß-catenin promoter region. Thus, we demonstrated that ATRA may reverse the side-effects of COX-2 inhibitor on bone metabolism through increasing the activation of Wnt/ß-catenin pathway partly.

17.
Gene ; 718: 144072, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31446095

RESUMO

Disorders of sex development (DSDs) are congenital conditions in which chromosomal, gonadal and sex is atypical. It is difficult to diagnose and manage patients with DSD in clinical practice, and the molecular etiology of DSD is still not completely understood. Here, we identified two novel pathogenic mutations from three unrelated Chinese patients with 46,XY complete gonadal dysgenesis (CGD) that is a clinical subgroup of DSD by whole exome sequencing. A novel mutation in the SRY gene (c.161delG) was identified in the first patient, and the second patient carried a novel missense mutation in the MAP3K1 gene (c.2117T>G). Bioinformatics analysis found that the deletion of SRY (c.161delG) led to a premature stop codon at amino acid 59 in the SRY protein, which resulted in lacking the DNA binding domain of SRY protein. Functional studies found that the missense mutation in the MAP3K1 gene (c.2117T>G) could interfere with the gene function through increasing the phosphorylation of the downstream targets of MAP3K1, ERK1/2 and p38, which resulted in reducing testis-determining factor SOX9 expression and increasing ovary-promoting factor ß-catenin activity. According to the American college of medical genetics and genomics (ACMG) standards and guidelines, these mutations were categorized as "pathogenic" mutations. Thus, our findings provide two novel pathogenic mutations associated with 46,XY CGD that can improve the etiological diagnosis for 46,XY CGD. ABBREVIATIONS.


Assuntos
Sequência de Bases , Disgenesia Gonadal 46 XY , MAP Quinase Quinase Quinase 1 , Mutação de Sentido Incorreto , Deleção de Sequência , Adulto , Grupo com Ancestrais do Continente Asiático , Feminino , Disgenesia Gonadal 46 XY/genética , Disgenesia Gonadal 46 XY/metabolismo , Disgenesia Gonadal 46 XY/patologia , Humanos , MAP Quinase Quinase Quinase 1/genética , MAP Quinase Quinase Quinase 1/metabolismo , Sistema de Sinalização das MAP Quinases/genética , Masculino , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Proteína da Região Y Determinante do Sexo/genética , Proteína da Região Y Determinante do Sexo/metabolismo
18.
FEBS Lett ; 593(19): 2706-2715, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31380564

RESUMO

Angiopoietins (Angs) are a family of vascular growth factors that share multiple cellular functions related to cell survival, proliferation, and migration. Angs play physiological and pathological roles through the Tie tyrosine kinase receptors. The Ang-Tie signaling pathway participates in the developmental and tumor-induced angiogenesis and is also involved in many disease settings, such as vascular diseases, systemic inflammation, and cancers. Since Angs are widely expressed in the kidney, an enormous amount of research focuses on their roles in the kidney. In this review, we describe the biological functions of the Ang-Tie signaling pathway and summarize their roles in kidney development and maturation, acute and chronic kidney diseases, diabetic nephropathy, lupus nephropathy, hemolytic uremic syndrome, end-stage renal diseases, and renal cell carcinoma. Understanding the molecular mechanisms of Ang-Tie signaling may reveal potential therapeutic targets for preventing or alleviating kidney diseases.

19.
ACS Synth Biol ; 8(9): 2092-2105, 2019 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-31465214

RESUMO

As an important post-transcriptional regulatory machinery mediated by ∼21nt short-interfering double-stranded RNA (siRNA), RNA interference (RNAi) is a powerful tool to delineate gene functions and develop therapeutics. However, effective RNAi-mediated silencing requires multiple siRNAs for given genes, a time-consuming process to accomplish. Here, we developed a user-friendly system for single-vector-based multiplex siRNA expression by exploiting the unique feature of restriction endonuclease BstXI. Specifically, we engineered a BstXI-based shotgun cloning (BSG) system, which consists of three entry vectors with siRNA expression units (SiEUs) flanked with distinct BstXI sites, and a retroviral destination vector for shotgun SiEU assembly. For proof-of-principle studies, we constructed multiplex siRNA vectors silencing ß-catenin and/or Smad4 and assessed their functionalities in mesenchymal stem cells (MSCs). Pooled siRNA cassettes were effectively inserted into respective entry vectors in one-step, and shotgun seamless assembly of pooled BstXI-digested SiEU fragments into a retroviral destination vector followed. We found these multiplex siRNAs effectively silenced ß-catenin and/or Smad4, and inhibited Wnt3A- or BMP9-specific reporters and downstream target expression in MSCs. Furthermore, multiplex silencing of ß-catenin and/or Smad4 diminished Wnt3A and/or BMP9-induced osteogenic differentiation. Collectively, the BSG system is a user-friendly technology for single-vector-based multiplex siRNA expression to study gene functions and develop experimental therapeutics.

20.
Gene ; 714: 143990, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31326550

RESUMO

BACKGROUND: Progressive cardiac conduction defect (PCCD), also known as Lenegre-Lev disease, is one of the most common heart conduction abnormalities. Previous studies have screened for known mutation sites that cause heart block in a 68-person family with a history of PCCD, revealed no mutations. OBJECTIVE: To screen pathogenic genes of the PCCD family and to study the function of the gene mutations related to heart block diseases. METHODS: Whole exome sequencing (WES) was performed on two PCCD patients and one non-PCCD family member to find the related pathogenic gene. After family co-segregation and preliminary functional analysis, we identified the mutant gene CLCA2. To study the function of this gene, we constructed mutant-gene mice using CRISPR-Cas9 technology, and electrocardiogram monitoring was performed after genotype verification. RESULTS: The CLCA2 c.G1725T mutation was identified and co-segregated with the phenotype. The analysis showed that the CLCA2 c.G1725T mutation is harmful and mainly affects protein glycosylation. Immunofluorescence staining revealed that CLCA2 was highly expressed in the sinoatrial node (SAN) tissues. Electrocardiogram monitoring of the mice revealed that CLCA2 point mutations induced mild conduction block and ectopic pacemakers. CONCLUSION: Our findings indicate that a novel heterozygous missense mutation c.G1725T of the CLCA2 gene may be associated with heart block disease and the mutation in this gene may lead to sinus node lesions and conduction blocking.


Assuntos
Canais de Cloreto/genética , Bloqueio Cardíaco/genética , Mutação de Sentido Incorreto/genética , Sequência de Aminoácidos , Animais , Eletrocardiografia/métodos , Feminino , Heterozigoto , Humanos , Masculino , Camundongos , Linhagem , Fenótipo , Mutação Puntual/genética , Nó Sinoatrial/patologia
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