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1.
Cell Prolif ; : e12738, 2020 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-31957155

RESUMO

OBJECTIVES: Podocyte injury is a prediction marker of diabetic nephropathy (DN), and AKT/mTOR pathway-mediated inhibition of autophagy is widely reported to contribute to podocyte damage. Recent study stated that sperm-associated antigen 5 (SPAG5) activated AKT/mTOR signalling in bladder urothelial carcinoma, indicating SPAG5 might regulate autophagy and play a role in podocyte damage. MATERIALS AND METHODS: Apoptosis and autophagy of human podocytes (HPCs) were detected by flow cytometry and immunofluorescence (IF). Gene level was assessed by Western blot and RT-qPCR. Molecular interactions were determined by pulldown, RNA immunoprecipitation (RIP), co-immunoprecipitation (co-IP), chromatin immunoprecipitation (ChIP) and luciferase reporter assays. RESULTS: SPAG5 mRNA and protein levels were upregulated under high glucose treatment in HPCs. Silencing SPAG5 reversed the increase of apoptosis and decrease of autophagy in high glucose-treated HPCs. Later, we found a long non-coding RNA (lncRNA) SPAG5 antisense RNA1 (SPAG5-AS1) as a neighbour gene to SPAG5. Mechanistically, YY1 transcriptionally upregulated SPAG5-AS1 and SPAG5 in high glucose-treated podocytes. SPAG5-AS1 acted as a competitive endogenous RNA (ceRNA) to regulate miR-769-5p/YY1 axis and induce SPAG5. SPAG5-AS1 interacted with ubiquitin-specific peptidase 14 (USP14) and leads to de-ubiquitination and stabilization of SPAG5 protein. CONCLUSIONS: This study revealed that SPAG5-AS1 inhibited autophagy and aggravated apoptosis of podocytes via SPAG5/AKT/mTOR pathway, indicating SPAG5-AS1/SPAG5 as a potential target for the alleviation of podocyte injury and offering new thoughts for the treatments of DN.

3.
Andrologia ; 52(1): e13449, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31657059

RESUMO

This retrospective study evaluated the correlation between the sperm DNA integrity results and infertile male age or sperm motility in 654 infertile men undergoing infertility evaluations from 2013 to 2016. The correlation between the results of sperm DNA integrity and male age was positive, while a negative correlation was detected between sperm DNA integrity and sperm motility in all subjects. According to age (≤30, 30-35 and ≥35), men with normozoospermia or abnormal semen parameters were, respectively, divided into groups 1, 2 and 3, or groups A, B and C. The sperm DNA fragmentation index (DFI) and DFI abnormality rates in groups 3 and C were highest among their respective cohorts. But they were not significantly different between groups within the same age range. Statistically significant differences were found in male age, progressive motility, as well as total motility between patients with normal DFIs and those with abnormal DFIs in group C, but not in group 3. Older (≥35 years) infertile men have increased sperm DNA fragmentation, independent of conventional semen parameters. Male age is more critical to sperm DNA integrity than routine semen parameters.

4.
J Clin Pharm Ther ; 2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31778586

RESUMO

WHAT IS KNOWN AND OBJECTIVE: Myelosuppression, an adverse drug reaction (ADR), often causes medical treatment termination in cancer patients. It has been known that genetic components, such as single-nucleotide polymorphisms (SNPs), influence the risk of myelosuppression at the individual-patient level. However, due to ethnic variation in frequency of genetic polymorphisms, results reported in Caucasian patients may not be generalizable to the Chinese Han population. Until now, few researches on myelosuppression included Chinese Han patients. In this study, we conducted a systematic study of potential biomarkers for docetaxel-induced myelosuppression in Han Chinese patients. METHODS: We examined 61 SNPs in 36 genes that code for drug transporters, metabolism enzymes, nuclear receptors and DNA repair pathway in 110 Chinese Han patients receiving docetaxel-based chemotherapy. Genotyping was conducted using the Sequenom MassARRAY system. Significant SNPs were identified by logistic regression, and gene-gene interactions were investigated by generalized multifactor dimensionality reduction (GMDR) analysis. RESULTS AND DISCUSSION: Our results revealed that 11 SNPs in nine genes (SLC15A1, SLCO1A2, CYP2D6, FMO3, UGT1A1, NAT2, SULT2A1, PXR and HNF4α) were associated with docetaxel-induced myelosuppression. GMDR analyses suggested that a 3-locus model: SLC15A1 rs2297322-PXR rs3732359-FMO3 rs2266782 was an appropriate predictive model of docetaxel-induced myelosuppression (P = .017, Testing Bal.Acc = 0.653, CV Consistency = 10/10). WHAT IS NEW AND CONCLUSION: Our findings suggest multiple novel predictive biomarkers of docetaxel-induced myelosuppression: SLC15A1 rs2297322, PXR rs3732359 and FMO3 rs2266782. These discoveries should help in advancing future personalized therapy of docetaxel-based chemotherapy specific to Chinese Han patients.

5.
BMC Pharmacol Toxicol ; 20(1): 66, 2019 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-31722728

RESUMO

BACKGROUND: Colchicine is a clinical medicine used for relief from gout and familial Mediterranean fever. Because of its toxic effects, intravenous injection of colchicine has been banned, but it is still widely administered orally. We assayed the toxic effects of colchicine in cultured primary chorionic villus cells and amniotic fluid cells to interpret its influence on the placenta and foetus. METHODS: Bright field record and cell count kit 8 were used to value cell viability. Flow cytometer was used to identify cells markers, cell cycle and cell apoptosis. G-banding was used for karyotype analysis for sample genetic and drug effect evaluation. RESULTS: Chorionic villus cells and amniotic fluid cells were characterized as mesenchymal cells that share most cell surface markers and have a similar response to colchicine. Colchicine did not induce a decline in cell viability at low concentrations but suppressed cell proliferation by arresting the cell cycle in the G2/M phase and increased the risk of tetraploid generation in a small subset of cases. CONCLUSIONS: Our study revealed the results of a colchicine-induced toxicity test in prenatal cells and determined the anti-mitotic biologically functional dose and manner of administration that might reduce the risk of tetraploid generation.

7.
Mol Genet Genomic Med ; 7(11): e963, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31566929

RESUMO

BACKGROUND: ß-thalassemia is one of the most common monogenic diseases in the world. Southeast China is a highly infected area affected by four ß-thalassemia mutation types (HBB:c.-78A>G, HBB:c.52A>T, HBB:c.126_129delCTTT, and HBB:c.316-197C>T). Relative haplotype dosage (RHDO), a haplotype-based approach, has shown promise as an application for noninvasive prenatal diagnosis (NIPD); however, additional family members (such as the proband) are required for haplotype construction. The abovementioned circumstances make RHDO-based NIPD cost prohibitive; additionally, the genetic information of the proband is not always available. Thus, it is necessary to find a practical method to solve these problems. METHODS: Targeted sequencing was applied to sequence parental genomic DNA and cell-free fetal DNA (cffDNA). Parental haplotypes were constructed with the SHAPEIT software based on the 1000 Genomes Project (1000G) Phase 3 v5 Southern Han Chinese (CHS) haplotype dataset. Single-nucleotide polymorphisms (SNPs) in the target region were called and classified, and the fetal mutation inheritance status was deduced using the RHDO method. RESULTS: Construction of the parental haplotypes and detection of the inherited parental mutations were successfully achieved in five families, despite a suspected recombination event. The status of the affected fetuses is consistent with the results of traditional reverse dot blot (RDB) diagnosis. CONCLUSION: This research introduced SHAPEIT into the classical RHDO workflow and proved that it is applicable to construct parental haplotypes without information from other family members.

8.
Biomed Pharmacother ; 120: 109481, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31586906

RESUMO

Glycyrrhizic acid ammonium salt (GAAS) is derived from glycyrrhizic acid, which is an active compound extracted from the Chinese traditional medicine licorice. GAAS is clinically applied to treat immune-mediated liver injury, but its mechanism remains elusive. Therefore, this study aimed to investigate the mechanism in which GAAS alleviates immune-mediated liver injury induced by Concanavalin A (ConA). After ten days of intragastric administration of GAAS, 20 mg/kg ConA was injected via tail vein to establish the immune-mediated liver injury model of BALB/C mice. Then, the concentrations of ALT, AST, and TBIL in the serum of mice were determined. H&E staining was performed to observe the pathological changes in the liver, and the expression of liver cytokines was detected by qPCR. Immunohistochemistry and Western blot analysis was employed to detect the expression of liver-related proteins. The apoptosis in liver tissue was detected by TUNEL. Our results suggest that GAAS demonstrated excellent protective effects in the liver. We found that GAAS down-regulated the mRNA expression of IL-1ß, IL-6, TNF-α, IFN-γ, and IL-17A, and it up-regulated the mRNA expression of IL-4 and TGF-ß. Additionally, GAAS may modulate the balance of four immune cells (Th1, Th2, Th17, and Treg) by regulating the expression of T-bet, GATA3, RORγt, and Foxp3 to alleviate liver injury in mice. Furthermore, GAAS decreased hepatocyte apoptosis by blocking the JAK1/STAT1/IRF1 pathway, suppressing oxidative stress, decreasing p-JNK expression, and regulating the expression of apoptosis-related proteins. In summary, the mechanism of GAAS in liver injury alleviation acts to regulate the balance of Th cells in the liver to inhibit hepatocyte apoptosis. This study may provide a new strategy for the treatment of immune-mediated liver injury.

9.
J Cell Mol Med ; 23(12): 8046-8057, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31631510

RESUMO

ß-thalassaemia is a prevalent hereditary haematological disease caused by mutations in the human haemoglobin ß (HBB) gene. Among them, the HBB IVS2-654 (C > T) mutation, which is in the intron, creates an aberrant splicing site. Bone marrow transplantation for curing ß-thalassaemia is limited due to the lack of matched donors. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9), as a widely used tool for gene editing, is able to target specific sequence and create double-strand break (DSB), which can be combined with the single-stranded oligodeoxynucleotide (ssODN) to correct mutations. In this study, according to two different strategies, the HBB IVS2-654 mutation was seamlessly corrected in iPSCs by CRISPR/Cas9 system and ssODN. To reduce the occurrence of secondary cleavage, a more efficient strategy was adopted. The corrected iPSCs kept pluripotency and genome stability. Moreover, they could differentiate normally. Through CRISPR/Cas9 system and ssODN, our study provides improved strategies for gene correction of ß-Thalassaemia, and the expression of the HBB gene can be restored, which can be used for gene therapy in the future.

10.
Biomed Res Int ; 2019: 6573497, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31119181

RESUMO

Background: Endocrinopathies are common in patients with ß-thalassemia major despite parenteral iron chelation therapy with deferoxamine. Prevalence of abnormal glucose metabolism in previous studies was controversial. The aim of this study was to discuss the prevalence of abnormal glucose metabolism in ß-thalassemia major based on a meta-analysis. Methods: PubMed, ScienceDirect, Springerlink, Ovid, Web of Science, MEDLINE, Wanfang database, and Chinese National Knowledge Internet were searched for relevant articles. Two authors selected the articles according to the inclusion criteria and then extracted the data. The prevalence of diabetes mellitus (DM) in ß-thalassemia major was defined as the primary outcome. The prevalence with the 95% confidence interval (95%CI) was used to evaluate the proportion of abnormal glucose metabolism and other endocrine disorders in patients with ß-thalassemia major. Subgroup analyses were applied to explore the prevalence in different regions. Sensitivity analysis and publication bias assessment were also conducted. Results: A total of 44 studies with 16605 cases were included in this analysis. Diabetes mellitus was present in 6.54% (95% CI: 5.30%-7.78%). The fixed subgroup study revealed that the region with the highest prevalence was the Middle East (prevalence= 7.90%, 95% CI: 5.75%-10.05%). The accumulated meta-analysis revealed that the prevalence of DM in ß-thalassemia major was relatively steady in each year. The prevalence of impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and other endocrine disorders in ß-thalassemia major was 17.21% (95% CI: 8.43%-26.00%), 12.46% (95% CI: 5.98%-18.94%), and 43.92% (95% CI: 37.94%-49.89%), respectively. Sensitivity analysis showed that the pooled results were robust; publication bias assessment revealed that there was no significant evidence that the pooled results were influenced by publication bias. Conclusion: High prevalence of endocrine disorders involving abnormal glucose metabolism was detected in ß-thalassemia major. Treatment and prevention measurements may be necessary to prevent growth and endocrine problems.


Assuntos
Diabetes Mellitus/epidemiologia , Doenças do Sistema Endócrino/epidemiologia , Glucose/metabolismo , Talassemia beta/epidemiologia , Terapia por Quelação , Desferroxamina/uso terapêutico , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patologia , Doenças do Sistema Endócrino/complicações , Doenças do Sistema Endócrino/metabolismo , Doenças do Sistema Endócrino/patologia , Intolerância à Glucose , Humanos , Quelantes de Ferro/uso terapêutico , Oriente Médio/epidemiologia , Talassemia beta/complicações , Talassemia beta/metabolismo , Talassemia beta/patologia
11.
Psych J ; 8(3): 353-362, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30932372

RESUMO

Repeated exposure to stress during early life has a lifetime impact on physical and psychological functions. Our study investigated the long-term effects of early life stress (ELS; measured retrospectively) on the hypothalamic-pituitary-adrenal (HPA) axis and sympathetic nervous system functions under psychosocial stress among healthy adults. Fifty healthy volunteers (33 male, 17 female, mean age 22.6 ± 1.8 years) underwent a standardized psychosocial stress protocol (the Trier Social Stress Test), with the collection of salivary cortisol, heart rates, and positive and negative affect. The results showed increases in cortisol, heart rate, and negative affect after the stress inducement. More importantly, a significant negative correlation was found between the severity of ELS and the increase of cortisol to the stress. The severity of stress in one's early life predicted his/her cortisol reactivity to the stress in adulthood. Neither the heart rate reactivity nor the affective reactivity shows significant association with ELS. The blunted cortisol reactivity reflects the alteration of the HPA axis, which may confer the risk for certain stress-related disorders.

12.
Plant Dis ; 103(4): 748-758, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30789316

RESUMO

Botrytis cinerea (anamorph of Botryotinia fuckeliana) causes gray mold on numerous plants, including kiwifruit. The primary aim of this study was to investigate the phenotypic and genetic characteristics of the Botrytis cinerea population from kiwifruit in Sichuan Province, China. In all, 176 isolates were collected from kiwifruit orchards from eight geographic regions in Sichuan. All isolates were identified as B. cinerea sensu stricto based on the combined datasets, including morphological criteria, determination of the Bc-hch allele, and phylogenetic analysis of the genes RPB2, G3PDH, and HSP60. Three colony types (i.e., sclerotial, mycelial, and conidial) were observed on potato dextrose agar after 2 weeks, with sclerotial isolates, the predominant category, accounting for 40.91%. No obvious differences in microscopic characteristics were observed among the three types. Three genotypes of transposable elements were identified in the B. cinerea population: boty, flipper, and transposa types. The most prevalent genotype from different geographic populations of B. cinerea was transposa; in contrast, the flipper genotype accounted for only 3.98% of the total population, whereas the vacuma genotype was absent. According to MAT locus amplification, 87 and 89 isolates are MAT1-1 and MAT1-2 type, respectively, and the two mating types were found to be balanced overall in the population. Forty-eight representative isolates were all able to cause gray mold to some extent, and disease severities were significantly different between the cultivars Hongyang and Hort16A (P < 0.01). Disease severity was significantly greater on young leaves than on mature leaves (P < 0.01). No significant relationship was found between pathogenicity and geographical region, colony type, or transposon distribution. The results obtained in the present study suggest a relatively uniform species diversity of Botrytis but rich phenotypic and genetic differentiation within the B. cinerea population on kiwifruit in China. Utilizing resistant cultivars and rain-shelter cultivation instead of fungicides may be an effective approach to delaying pathogen variability.


Assuntos
Actinidia , Botrytis , Actinidia/microbiologia , Botrytis/classificação , Botrytis/genética , China , Filogenia , Doenças das Plantas/microbiologia
13.
Cell Biosci ; 9: 16, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30774927

RESUMO

Background: Neoadjuvant chemotherapy (NAC) induces a pathological complete response (pCR) in ~ 30% of patients with breast cancer. However, aberrant DNA methylation alterations are frequent events during breast cancer progression and acquisition of chemoresistance. We aimed to characterize the inter- and intra-tumor methylation heterogeneity (MH) in breast cancer following NAC. Methods: DNA methylation profiles of spatially separated regions of breast tumors before and after NAC treatment were investigated using high-density methylation microarray. Methylation levels of genes of interest were further examined using multiplexed MethyLight droplet digital PCR (ddPCR). Results: We have discovered different levels of intra-tumor MH in breast cancer patients. Moreover, NAC dramatically altered the methylation profiles and such changes were highly heterogeneous between the patients. Despite the high inter-patient heterogeneity, we identified that stem cell quiescence-associated genes ALDH1L1, HOPX, WNT5A and SOX9 were convergently hypomethylated across all the samples after NAC treatment. Furthermore, by using MethyLight ddPCR, we verified that the methylation levels of these 4 genes were significantly lower in breast tumor samples after NAC than those before NAC. Conclusions: Our study has revealed that NAC dramatically alters epigenetic heterogeneity in breast cancer and induces convergent hypomethylation of stem cell quiescence-associated genes, ALDH1L1, HOPX, WNT5A and SOX9, which can potentially be developed as therapeutic targets or biomarkers for chemoresistance.

14.
Stem Cell Res ; 34: 101379, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30605838

RESUMO

EDA is a gene located at Xq13.1. It encodes different isoforms of tumor necrosis factor (TNF) superfamily member ectodysplasin A. Ectodysplasin A is a transmembrane protein which can be cleaved to form a secreted form and interact with EDA receptor to mediate the development of ectoderm. Mutations of the EDA gene are related to ectodermal dysplasia and tooth agenesis. Here, we report the establishment of the EDA gene knockout human embryonic stem (hES) cell line by CRISPR-Cas9 technology. This cell line provides good materials for further studies of the roles ectodysplasin A plays in ectoderm differentiation and tooth development.


Assuntos
Sistemas CRISPR-Cas/genética , Técnicas de Cultura de Células/métodos , Displasia Ectodérmica/genética , Displasia Ectodérmica/patologia , Técnicas de Inativação de Genes , Células-Tronco Embrionárias Humanas/citologia , Linhagem Celular , Humanos , Masculino
15.
Biomed Pharmacother ; 111: 740-750, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30611999

RESUMO

Albeit microRNAs (miRNAs) have become increasingly appreciated for their essential roles in innate immune responses to viral infections; however, it is unknown how host miRNAs regulate influenza A virus (IAV)-induced inflammation. The aim of our study was to investigate the role of miR-146a in IAV replication in vitro and in vivo. In vitro, we found miR-146a was significantly upregulated in A549 cells with IAV infection. Overexpression of miR-146a promoted IAV replication, while downregulation of miR-146a repressed replication. We found that miR-146a diminished type I interferon (IFN) responses by decreasing IFN-ß production and IFN-stimulated gene (ISG) expression. Furthermore, we found the IFNs level and IAV replication regulated by miR-146a inhibitor was partially reversed by depletion of interferon receptor (IFNAR) 1 or 2. In addition, we found that miR-146a directly targets tumor necrosis factor receptor association factor 6 (TRAF6), which is involved in the production of type I IFN, and TRAF6 overexpression reversed the replication-promoting effect of miR-146a on IAV. In vivo, inhibition of miR-146a alleviated IAV-induced mice lung injury and promoted survival rates by promoting type I antiviral activities. It is, therefore, concluded that downregulation of miR-146a inhibits IAV replication by enhancing type I IFN response through its target gene TRAF6 in vitro and in vivo, suggesting miR-146a antagomir might be a potential therapeutic target during IAV infection.


Assuntos
Regulação para Baixo/fisiologia , Vírus da Influenza A/metabolismo , Interferon Tipo I/farmacologia , MicroRNAs/metabolismo , Replicação Viral/fisiologia , Células A549 , Animais , Antagomirs/farmacologia , Regulação para Baixo/efeitos dos fármacos , Células HEK293 , Humanos , Vírus da Influenza A/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/antagonistas & inibidores , Distribuição Aleatória , Células Vero , Replicação Viral/efeitos dos fármacos
16.
Medicine (Baltimore) ; 97(40): e12507, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30290606

RESUMO

INTRODUCTION: Phosphaturic mesenchymal tumor mixed connective tissue type (PMT/MCT) is the most common type (up to 90%) of phosphaturic mesenchymal tumor (PMT), a rare clinicopathologic entity. Besides overproduction of fibroblast growth factor 23 (FGF23), there is a big variation of immunohistochemical characteristic across types of PMT, which makes it difficult to obtain an early diagnosis of PMT/MCT. As a benign tumor, PMT/MCT usually happens in subcutaneous tissues and leads to nonhealing of wound. A complete excision of PMT/MCT facilitates wound healing. CONCLUSIONS: Review of the existing evidence indicates that early diagnosis of PMT/MCT is critically important when treating PMT/MCT wound. Hence standardization of early diagnosis for PMT/MCT is mandated.


Assuntos
Hipofosfatemia Familiar/diagnóstico , Mesenquimoma/diagnóstico , Doença Mista do Tecido Conjuntivo/diagnóstico , Neoplasias de Tecidos Moles/diagnóstico , Ferimentos e Lesões/complicações , Biomarcadores Tumorais/sangue , Diagnóstico Diferencial , Detecção Precoce de Câncer , Fatores de Crescimento de Fibroblastos/biossíntese , Humanos , Hipofosfatemia Familiar/sangue , Hipofosfatemia Familiar/etiologia , Mesenquimoma/sangue , Mesenquimoma/etiologia , Doença Mista do Tecido Conjuntivo/sangue , Doença Mista do Tecido Conjuntivo/etiologia , Neoplasias de Tecidos Moles/sangue , Neoplasias de Tecidos Moles/etiologia , Ferimentos e Lesões/sangue
17.
Mol Cell ; 72(2): 380-394.e7, 2018 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-30293782

RESUMO

RNA splicing is a critical mechanism by which to modify transcriptome, and its dysregulation is the underlying cause of many human diseases. It remains challenging, however, to genetically modulate a splicing event in its native context. Here, we demonstrate that a CRISPR-guided cytidine deaminase (i.e., targeted-AID mediated mutagenesis [TAM]) can efficiently modulate various forms of mRNA splicing. By converting invariant guanines to adenines at either 5' or 3' splice sites (SS), TAM induces exon skipping, activation of alternative SS, switching between mutually exclusive exons, or targeted intron retention. Conversely, TAM promotes downstream exon inclusion by mutating cytidines into thymines at the polypyrimidine tract. Applying this approach, we genetically restored the open reading frame and dystrophin function of a mutant DMD gene in patient-derived induced pluripotent stem cells (iPSCs). Thus, the CRISPR-guided cytidine deaminase provides a versatile genetic platform to modulate RNA splicing and to correct mutations associated with aberrant splicing in human diseases.


Assuntos
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Citidina Desaminase/genética , Processamento de RNA/genética , Sequência de Aminoácidos , Animais , Linhagem Celular , Distrofina/genética , Éxons/genética , Redes Reguladoras de Genes , Células HEK293 , Humanos , Íntrons/genética , Camundongos , Fases de Leitura Aberta/genética , Sítios de Splice de RNA/genética
18.
MAbs ; 10(8): 1301-1311, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30204048

RESUMO

Animal models used to evaluate efficacies of immune checkpoint inhibitors are insufficient or inaccurate. We thus examined two xenograft models used for this purpose, with the aim of optimizing them. One method involves the use of peripheral blood mononuclear cells and cell line-derived xenografts (PBMCs-CDX model). For this model, we implanted human lung cancer cells into NOD-scid-IL2Rg-/- (NSI) mice, followed by injection of human PBMCs. The second method involves the use of hematopoietic stem and progenitor cells and CDX (HSPCs-CDX model). For this model, we first reconstituted the human immune system by transferring human CD34+ hematopoietic stem and progenitor cells (HSPCs-derived humanized model) and then transplanted human lung cancer cells. We found that the PBMCs-CDX model was more accurate in evaluating PD-L1/PD-1 targeted immunotherapies. In addition, it took only four weeks with the PBMCs-CDX model for efficacy evaluation, compared to 10-14 weeks with the HSPCs-CDX model. We then further established PBMCs-derived patient-derived xenografts (PDX) models, including an auto-PBMCs-PDX model using cancer and T cells from the same tumor, and applied them to assess the antitumor efficacies of anti-PD-L1 antibodies. We demonstrated that this PBMCs-derived PDX model was an invaluable tool to study the efficacies of PD-L1/PD-1 targeted cancer immunotherapies. Overall, we found our PBMCs-derived models to be excellent preclinical models for studying immune checkpoint inhibitors.


Assuntos
Antígeno B7-H1/antagonistas & inibidores , Imunoterapia/métodos , Leucócitos Mononucleares/imunologia , Neoplasias Pulmonares/terapia , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Ensaios Antitumorais Modelo de Xenoenxerto , Animais , Antígeno B7-H1/imunologia , Antígeno B7-H1/metabolismo , Linhagem Celular Tumoral , Células Cultivadas , Modelos Animais de Doenças , Células-Tronco Hematopoéticas/imunologia , Células-Tronco Hematopoéticas/metabolismo , Humanos , Leucócitos Mononucleares/metabolismo , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/metabolismo , Camundongos Endogâmicos NOD , Camundongos Knockout , Camundongos SCID , Receptor de Morte Celular Programada 1/imunologia , Receptor de Morte Celular Programada 1/metabolismo
19.
Biochem Biophys Res Commun ; 505(1): 45-50, 2018 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-30236984

RESUMO

The liver is a central metabolic organ. Activating transcription factor 6 (ATF6) acts as an ER stress responsive gene and is reported to attenuate hepatic steatosis. Over expressing a dominant-negative form of ATF6 exacerbates glucose intolerance and insulin resistance. In the present study, we used the conditional knockout technique to specifically knockout ATF6 in the mouse liver. We used qPCR to detect the mRNA levels of related genes. Western blot analysis was used to evaluate protein levels. Flow cytometry assay showed the apoptosis status. Glucose tolerance tests and insulin tolerance tests were used to determine glucose and insulin sensitivity. The results showed that liver specific knockout of ATF6 exacerbated HFD-induced hepatic steatosis and glucose tolerance. Abolished ATF6 exacerbated gluconeogenic metabolism by MTOR mediated down regulation of autophage. In conclusion, these findings suggest that therapeutic strategies by supplementing ATF6 may be beneficial for the treatment of glucose intolerance as well as insulin resistance in the high fat induced liver metabolic damage condition.


Assuntos
Fator 6 Ativador da Transcrição/metabolismo , Autofagia , Fígado/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Fator 6 Ativador da Transcrição/genética , Animais , Dieta Hiperlipídica/efeitos adversos , Fígado Gorduroso/etiologia , Fígado Gorduroso/genética , Fígado Gorduroso/metabolismo , Gluconeogênese/genética , Teste de Tolerância a Glucose , Resistência à Insulina/genética , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transdução de Sinais , Serina-Treonina Quinases TOR/genética
20.
Mol Med Rep ; 18(2): 1981-1986, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29901147

RESUMO

The aim of the present study was to verify the effects of heavy metal coupling agents (sodium citrate and EDTA) and antioxidants (acetyl carnitine and lipoic acid) on the number of oocytes, as well as the ageing of mitochondria, chromosomes and spindles in mice. C57BL/6 female mice were randomly classified into four groups (n=12 per group): i) Heavy metal coupling agent; ii) antioxidant; iii) mixed group; and iv) the normal control group. For the treatments, heavy metal coupling agents and antioxidants were added to the drinking water provided to the mice. Following 3, 6, 9 and 12 months of treatment, the number of oocytes and mitochondrial membrane potential were determined, and chromosome and spindle structures were observed. With increasing age, the experimental mice in the four groups showed significantly decreased numbers of oocytes, reduced mitochondrial activity, and increased rates of spindle and chromosome abnormalities, which indicated age­induced ageing of mouse oocytes; thus, a mouse ageing oocyte model had been successfully established. For mice of the same age, more oocytes, higher mitochondrial activity, and lower spindle and chromosome malformation rates were detected in the antioxidant and mixed groups when compared with the normal control groups. Furthermore, no significant difference in the number of oocytes, mitochondrial activity or chromosome malformation rates was observed between the heavy metal coupling agent group and normal control group, which was possibly due to less metal being absorbed during the breeding process. Therefore, the results demonstrated that the antioxidants acetyl carnitine and lipoic acid may serve a role in delaying oocyte ageing.


Assuntos
Antioxidantes/metabolismo , Senescência Celular , Aberrações Cromossômicas , Cromossomos de Mamíferos/metabolismo , Potencial da Membrana Mitocondrial , Membranas Mitocondriais/metabolismo , Oócitos/metabolismo , Animais , Feminino , Camundongos , Membranas Mitocondriais/patologia
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