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1.
Zhongguo Zhong Yao Za Zhi ; 46(20): 5382-5392, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34738442

RESUMO

To investigate the changes of bile acid(BA) levels in mice with sleep deprivation and the regulatory effect of Jiaotai Pills(JTP) on bile acid metabolism, this study established an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for simultaneous determination of 23 BAs in mice. A total of 24 ICR mice were randomized into normal group, model group, and JTP group. Mice in the model group and JTP group were deprived of sleep at 20 h·d~(-1) by sleep deprivation apparatus for 8 consecutive days. Mice in the JTP group were given(ig, qd) JTP 3.3 g·kg~(-1) and those in the normal group and model group received(ig) the same volume of purified water. UPLC conditions are as follows: Waters ACQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 µm), gradient elution with the mobile phase of 0.1% formic acid in water-methanol. MS conditions are as below: negative-ion electrospray ionization, multiple reaction monitoring(MRM). Thereby, the content of 23 BAs in serum, liver, and ileum was determined and methodological investigation of the method was performed. The results showed that 23 BAs could be accurately determined within 15 min and the correlation coefficients were all higher than 0.99. The precision, accuracy, specificity, reproducibility, matrix effect, and recovery of BAs all met the requirement. The levels of BAs were significantly increased in the serum, liver, and ileum of sleep-deprived mice, but JTP can significantly reduce the levels. The UPLC-MS/MS method is simple, rapid, and accurate, which can be used for the determination of 23 BAs in biological samples, and JTP can adjust the elevated BA levels of sleep-deprived mice.


Assuntos
Ácidos e Sais Biliares , Espectrometria de Massas em Tandem , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Medicamentos de Ervas Chinesas , Camundongos , Camundongos Endogâmicos ICR , Reprodutibilidade dos Testes , Sono
2.
J Cancer ; 12(23): 6979-6988, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34729100

RESUMO

Increasing evidence indicates that Notch signaling regulates multiple intracellular biological processes in malignant melanoma. Whereas how Notch signaling is transduced to influence melanoma cell behaviors remains largely elusive. Here we show that the Notch signaling downstream target Hey1 promotes migration and invasion of melanoma cells via the GRB2/PI3K/AKT pathway. First, bioinformatics tools, immunohistochemistry, and Western blotting analysis showed that the expression of Hey1 is increased in melanoma. Then, both in vivo and in vitro experiments showed that Hey1 promotes the malignant behaviour of the melanoma cells. High-throughput RNA-sequencing analysis revealed that inhibition of Hey1 results in decreased GRB2 expression in melanoma cells. Last, functional experiments confirmed that Hey1 positively regulates GRB2/PI3K/AKT pathway to influence migration and invasion of melanoma cells. In summary, our results suggest that Hey1 promotes the invasion and metastasis of melanoma cells by regulating GRB2/PI3K/AKT pathway. Our study provides potential therapeutics in tumor biology.

3.
Artigo em Inglês | MEDLINE | ID: mdl-34783430

RESUMO

Infrared nonlinear optical (IR NLO) crystals play a significant role in the development of IR laser technology. But rationally designing a new IR NLO crystal remains a huge challenge because of the unpredictability of inorganic structures. Herein, inspired by phase transformation of the famous BaB2 O4 (BBO) crystal, a temperature-induced centrosymmetric (CS) to noncentrosymmetric (NCS) transformation approach is employed in CS BaGa2 Se4 to uncover a new NCS ternary chalcogenide, ß-BaGa2 Se4 which can exhibit the well-balanced NLO properties, including moderate phase-matching SHG response (≈0.6×AgGaS2 ), high LDT (10×AgGaS2 ), and large birefringence (Δn=0.18 in the visible region). Therefore, ß-BaGa2 Se4 will be a promising IR NLO crystal. The analysis for the structure-property relationship shows that the excellent NLO properties of ß-BaGa2 Se4 mainly originate from edge-sharing GaSe4 tetrahedral chains, which are different from B3 O6 groups in ß-BBO and respect a new direction for the design of IR NLO crystals. So, we believe that the current research provides not only a new IR NLO crystal but also some insights for the design of new IR NLO crystals.

4.
J Colloid Interface Sci ; 608(Pt 2): 1355-1365, 2021 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-34742058

RESUMO

Mitochondria are appealing targets in cancer therapy for providing a suitable microenvironment and energy supply. Herein, we constructed a glycosylated poly(amido amine)/celastrol (PAMAM/Cel) complex for hypoxia-activated mitochondria-specific drug delivery and chemothermal therapy to inhibit tumor growth and metastasis. The complex was characterized by high photothermal conversion efficiency, hypoxia-sensitive polyethylene glycol (PEG) outer layer detachment, and alkaline-sensitive drug release. The complex showed specific cellular uptake in glucose transporter 1 (GLUT1)-overexpressing tumor cells and mitochondrial accumulation in a hypoxic environment. Combined with near-infrared (NIR) laser irradiation, the complex exhibited higher cytotoxicity, apoptosis induction, and metastasis inhibition rates due to the synergistic chemothermal effect. Similarly, the complex also targeted tumors and accumulated in mitochondria in tumor-bearing nude mice, resulting in superior inhibitory effects on tumor growth and metastasis as well as low systematic toxicity. Further mechanistic studies discovered that the complex impaired the mitochondrial membrane, reduced adenosine triphosphate (ATP) content, and regulated metastasis-related protein expression. Thus, the present study provides a promising nanomedicine for tumor therapy.

5.
J Clin Transl Hepatol ; 9(5): 702-710, 2021 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-34722185

RESUMO

Background and Aims: The expression of discoidin domain receptor 1 (DDR1) is commonly up-regulated and undergoes collagen-induced ectodomain (N-terminal) shedding during the progression of liver fibrosis. This study aimed to evaluate the clinical utility of N-terminal DDR1 as a diagnostic biomarker for liver fibrosis. Methods: N-terminal DDR1 shedding was evaluated using cell lines, liver fibrosis mouse models, clinical data of 298 patients collected from February 2019 to June 2020. The clinical data were divided into test and validation cohorts to evaluate the diagnostic performance of serum N-terminal DDR1. Results: Time- and dosage-dependent N-terminal DDR1 shedding stimulated by collagen I was observed in a hepatocyte cell line model. The type I collagen deposition and serum N-terminal DDR1 levels concurrently increased in the development of liver fibrosis in mouse models. Clinical data demonstrated a significant diagnostic power of serum N-terminal DDR1 levels as an accurate biomarker of liver fibrosis and cirrhosis. The diagnostic performance was further increased when applying N-DDR1/albumin ratio, achieving area under the curve of 0.790, 0.802, 0.879, and 0.865 for detecting histological fibrosis stages F ≥2, F ≥3, F 4 with liver biopsy as a reference method, and cirrhosis according to imaging techniques, respectively. With a cut-off of 55.6, a sensitivity, specificity, positive predictive value, and negative predictive value of 82.7%,76.6%, 67.4%, and 88.3% were achieved for the detection of cirrhosis. Conclusions: Serum N-terminal DDR1 appears to be a novel diagnostic marker for liver fibrosis.

7.
Artigo em Inglês | MEDLINE | ID: mdl-34843047

RESUMO

The study aims to empirically estimate the nexus of green bond financing with renewable energy index OECD countries. Using the OECD countries data over the period of the 2011-2019, the study estimated the nexus between constructs. To justify the study findings and present widespread policy implications on recent topicality Padroni unit root test, FMOLS and DOLS technique is applied. For robustness analysis, long-run sensitivity analysis using FMOLS extension is used, and a comparative picture of green bond financing nexus with renewable energy index is presented. The study presented the consistent effects of green bond financing on renewable energy index indicators. This asymmetrical role of green bonds is confirmed on renewable energy indicators over the sample period. OECD countries injected 31% role of green bond financing on renewable energy index constructs, and it raised 9.4% of per unit energy efficiency in renewable energy systems; by this, the study findings warrant maximum support through public office, energy ministries, and departments for energy efficiency optimization. The study presents multiple policy implications to enhance renewable energy generation for energy efficiency through different alternative sources. Despite growing literature, the empirical discussion on this topicality is still shattered and less studied, which is extended and contributed by recent research. Furthermore, efficient regulation in the renewable energy sector may convert financial uncertainty into a huge opportunity. Investing in renewable energy stocks might help investors diversify their portfolios.

8.
Artigo em Inglês | MEDLINE | ID: mdl-34600150

RESUMO

PURPOSE: This study aimed to quantitatively evaluate whether the severity of menton deviation (MD) influenced facial perceptions of laypeople. We also aimed to determine the effectiveness of surgery in normalizing the distribution of the facial visual attention of laypeople. METHODS: The scanning paths of 177 laypeople were recorded using an eye tracking device while observing images of individuals without MD and pre- and post-treatment subjects with different degrees of MD. The fixation durations on the areas of interest (AOIs) in each group were compared and analysed. RESULTS: When observing the images of non-MD subjects, the eyes were the focus of the most significant fixation (higher than the fixations on the nose and lower face). When the MD increased to 3°, attention on the lower face increased (p = 0.001) with decreased attention to the eyes (p = 0.0126). At an MD of 9°, attention to the lower face sharply increased, even more so than that to the eyes, with decreased attention to the nose (p = 0.0104). Compared with the findings for the post-treatment images, the laypeople who observed the pretreatment images focused longer on the lower face and less on the eyes and nose (p = 0.001, p = 0.0322 and p = 0.0023, respectively). The distribution of the fixation duration when observing the post-treatment images was similar to that when observing the images of the non-MD subjects. CONCLUSIONS: Laypeople can perceive an MD of 3°, which causes changes in the distribution of visual attention, with attention focusing on the MD. When the deviation reaches 9°, it is very noticeable. Surgery can normalize the distribution of the facial visual attention of laypeople, as shown by the responses to the post-treatment images.

10.
Artigo em Inglês | MEDLINE | ID: mdl-34664060

RESUMO

The dynamic and reversible regulation roles of m6A modification and the characterization of m6A readers have provided new insights into spermatogenesis at the post-transcriptional level. YTHDF2, as an m6A reader, has been reported to mediate the m6A-containing transcript decay during the mouse oocyte maturation, embryonic stem cell differentiation, neural development, and zebrafish maternal-to-zygotic transition. However, the roles of YTHDF2 in mammalian spermatogenesis are uncertain. Here, we generated germ cell-specific Ythdf2 mutants (Ythdf2-vKO) at a C57BL/6J background and demonstrated that YTHDF2 is essential for mouse spermatogenesis and fertility. Ythdf2-vKO provides oligoasthenoteratozoospermia phenotype with increased apoptosis in germ cells. High-throughput RNA-seq analysis showed that a group of mRNAs is upregulated in Ythdf2-vKO mouse testis; further analysis and MeRIP-qPCR data showed that most of the upregulated genes in Ythdf2-vKO mouse testis are modified with m6A and are YTHDF2 candidate binding genes. Interestingly, RNA-seq analysis combined with our previous single-cell transcriptomics data of mouse spermatogenesis pointed out the failure of a wave of transcript transition during the spermatogenesis of Ythdf2-vKO mice, which was confirmed by gene expression analysis using qPCR of diplotene spermatocytes and round spermatids obtained through fluorescence-activated cell sorting. Our study demonstrates the fundamental role of YTHDF2 during mouse spermatogenesis and provides a potential candidate for the diagnosis of male infertility with the oligoasthenoteratozoospermia syndrome.

11.
Front Vet Sci ; 8: 686609, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34604366

RESUMO

Folic acid is a water-soluble B vitamin, and plays an important role in regulating gene expression and methylation. The liver is the major site of lipid biosynthesis in the chicken. Nevertheless, how gene expression and regulatory networks are affected by folic acid in liver of broilers are poorly understood. This paper conducted the RNA-seq technology on the liver of broilers under folic acid challenge investigation. First, 405 differentially expressed genes (DEGs), including 157 significantly upregulated and 248 downregulated, were detected between the control group (C) and the 5 mg folic acid group (M). Second, 68 upregulated DEGs and 142 downregulated DEGs were determined between C group and 10 mg folic acid group (H). Third, there were 165 upregulated genes and 179 downregulated genes between M and H groups. Of these DEGs, 903 DEGs were successfully annotated in the public databases. The functional classification based on GO and KEEGG showed that "general function prediction only" represented the largest functional classes, "cell cycle" (C vs. M; M vs. H), and "neuroactive ligand-receptor interaction" (C vs. H) were the highest unique sequences among three groups. SNP analysis indicated that numbers of C, M and H groups were 145,450, 146,131, and 123,004, respectively. Total new predicted alternative splicing events in C, M, and H groups were 9,521, 9,328, and 8,929, respectively. A protein-protein interaction (PPI) network was constructed, and the top 10 hub genes were evaluated among three groups. The results of real time PCR indicated that mRNA abundance of PPARγ and FAS in abdominal fat of M and H groups were reduced compared with the C group (P < 0.05). Ultramicroscopy results showed that folic acid could reduce lipid droplets in livers from chickens. Finally, contents of LPL, PPARγ, and FAS in abdominal fat were decreased with the folic acid supplmented diets (P < 0.01). These findings reveal the effects of folic acid supplemention on gene expression in liver of broilers, which can provide information for understanding the molecular mechanisms of folic acid regulating liver lipid metabolism.

12.
Oxid Med Cell Longev ; 2021: 9955717, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34650666

RESUMO

Dynamic cytoskeletal rearrangements underlie the changes that occur during cell division in proliferating cells. MICAL2 has been reported to possess reactive oxygen species- (ROS-) generating properties and act as an important regulator of cytoskeletal dynamics. However, whether it plays a role in gastric cancer cell proliferation is not known. In the present study, we found that MICAL2 was highly expressed in gastric cancer tissues, and this high expression level was associated with carcinogenesis and poor overall survival in gastric cancer patients. The knockdown of MICAL2 led to cell cycle arrest in the S phase and attenuated cell proliferation. Concomitant with S-phase arrest, a decrease in CDK6 and cyclin D protein levels was observed. Furthermore, MICAL2 knockdown attenuated intracellular ROS generation, while MICAL2 overexpression led to a decrease in the p-YAP/YAP ratio and promoted YAP nuclear localization and cell proliferation, effects that were reversed by pretreatment with the ROS scavenger N-acetyl-L-cysteine (NAC) and SOD-mimetic drug tempol. We further found that MICAL2 induced Cdc42 activation, and activated Cdc42 mediated the effect of MICAL2 on YAP dephosphorylation and nuclear translocation. Collectively, our results showed that MICAL2 has a promotive effect on gastric cancer cell proliferation through ROS generation and Cdc42 activation, both of which independently contribute to YAP dephosphorylation and its nuclear translocation.

13.
World J Clin Cases ; 9(27): 8268-8273, 2021 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-34621890

RESUMO

BACKGROUND: Major hip surgery usually requires neuraxial or general anesthesia with tracheal intubation and may be supplemented with a nerve block to provide intraoperative and postoperative pain relief. CASE SUMMARY: This report established that hip surgical procedures can be performed with a fascia iliaca compartment block (FICB) and monitored anesthesia care (MAC) while avoiding neuraxial or general anesthesia. This was a preliminary experience with two geriatric patients with hip fracture, American Society of Anesthesiologists status III, and with many comorbidities. Neither patient could be operated on within 48 h after admission. Both general anesthesia and neuraxial anesthesia were high-risk procedures and had contraindications. Hence, we chose nerve block combined with a small amount of sedation. Intraoperative analgesia was provided by single-injection ultrasound-guided FICB. Light intravenous sedation was added. Surgical exposure was satisfactory, and neither patient complained of any symptoms during the procedure. CONCLUSION: This report showed that hip surgery for geriatric patients can be performed with FICB and MAC, although complications and contraindications are common. The anesthetic program was accompanied by stable respiratory and circulatory system responses and satisfactory analgesia while avoiding the adverse effects and problems associated with either neuraxial or general anesthesia.

14.
Brain Imaging Behav ; 2021 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-34718941

RESUMO

This study aimed to investigate alterations of brain functional network connectivity (FNC) in lung cancer patients after chemotherapy and explore links between these FNC differences and cognitive impairment. Twenty-two lung cancer patients receiving chemotherapy and 26 healthy controls (HCs) underwent resting-state functional MRI (rs-fMRI) and neuropsychological testing. Group independent component analysis (GICA) was applied to rs-fMRI data to extract whole-brain resting state networks (RSNs). Static and dynamic FNC (dFNC) were constructed to reveal RSNs connectivity alterations between lung cancer patients and HCs group, and the correlations between the group differences in RSNs and cognitive performance were analyzed. Our findings revealed that chemotherapeutics can produce widespread connectivity abnormalities in RSNs, mainly focused on default mode network (DMN) and executive control network. Furthermore, the dFNC analysis help identify network configurations of each state and capture more chemotherapy-induced disorders of interactions between and within RSNs, which mainly includes sensorimotor network, attentional network and auditory network. In addition, after chemotherapy, the lung cancer patients spend shorter mean dwell time (MDT) in state 2. The decreased dFNC between DMN [independent component 5 (IC5)] and DMN (IC6) in the lung cancer patients after chemotherapy in state 4 was negatively correlated with Montreal Cognitive Assessment (MoCA) scores (r=-0.447, p=0.042). The dFNC analysis enrich our understanding of the neural mechanisms underlying the chemobrain, and suggested that the temporal dynamics of FNC could be a potential effective method to detect cognitive changes in lung cancer patients receiving chemotherapy.

15.
Sheng Li Xue Bao ; 73(5): 821-827, 2021 Oct 25.
Artigo em Chinês | MEDLINE | ID: mdl-34708239

RESUMO

ß3-adrenergic agonists induce adaptive thermogenesis and promote beiging of white fat. However, it remains unclear which metabolites mediate the stimulatory effects of ß3-adrenergic agonists on thermogenesis of brown and beige fat. In this study, adipose tissue was isolated from 8-week-old C57/BL6J male mice by intraperitoneal administration of ß3-adrenergic agonist CL316,243 for RNA-Seq, which revealed that histidine decarboxylase, a key enzyme in histamine synthesis, was strongly induced in adipose by CL316,243. Therefore, we speculated that histamine might be involved in the process of thermogenesis in adipose tissue. We determined the physiological role and mechanism by which histamine promotes fat thermogenesis by intravenous administering histamine to C57BL/6J mice fed a normal or a high-fat diet. The results showed that intravenous injection of histamine into C57BL/6J mice fed a normal diet stimulated the expression of thermogenic genes, including peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) and uncoupling protein 1 (UCP1), in brown adipose tissue (BAT) and inguinal white adipose tissue (iWAT). H&E staining also suggested that histamine treatment decreased the size of lipid droplets in adipocytes. Moreover, histamine treatment also enhanced thermogenesis of fat in high-fat diet induced obese mice, and improved glucose intolerance and fatty liver phenotype. Finally, we demonstrated that the effects of histamine on the thermogenic program were cell autonomous. Our data suggest that histamine may mediate the effects of ß3-adrenergic agonists on thermogenesis of fat.


Assuntos
Tecido Adiposo Bege , Histamina , Tecido Adiposo Marrom , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Termogênese , Proteína Desacopladora 1/genética
16.
Lab Chip ; 21(18): 3412-3419, 2021 09 14.
Artigo em Inglês | MEDLINE | ID: mdl-34472548

RESUMO

Droplet microfluidics has recently emerged as a powerful platform for a variety of biomedical applications including microreactors, bioactive compound encapsulation, and single cell culture and analysis; all these applications require long-term droplet stability, which, however, makes breaking the emulsion and retrieving the loaded samples difficult. Herein, we developed a novel class of thermo-responsive fluorosurfactants to control the droplet status simply by temperature. The surfactants were synthesized by coupling perfluorinated polyethers (PFPEs) with a thermo-responsive block of poly(N-isopropylacrylamide) (pNIPAM) or poly(2-ethyl-2-oxazoline) (pEtOx) with lower critical solution temperature (LCST). These diblock surfactants can stabilize the emulsion at temperatures below LCST due to the hydrophilic head, which became hydrophobic upon increasing the ambient temperature above LCST, thereby destabilizing the droplets and realizing demulsification simply via temperature control. The diblock surfactant can be applied for templating cell encapsulation using alginate microgels, which allowed one-step and high-throughput microfluidic generation of cell-laden microgels without compromising cell viability. This non-invasive, on-demand demulsification strategy provides a high degree of freedom for microencapsulation and on-demand recovery of the samples or reaction products within the droplets, which opens a new avenue for a wide range of applications of droplet-templating microfluidics.


Assuntos
Microfluídica , Tensoativos , Alginatos , Emulsões , Temperatura
17.
J Cosmet Dermatol ; 2021 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-34582097

RESUMO

BACKGROUND: Rosacea is a chronic inflammatory skin disease that affects people's life quality. It has been found to be related to many detrimental factors including ultraviolet exposure. However, the association between alcohol consumption and rosacea has long been debated. AIMS: To elucidate this association, we conducted a systematic review and meta-analysis. METHODS: We performed a systematic search of the literature published before February 16, 2021 on PubMed, Embase, and the Cochrane database and used a meta-analytic approach to calculate the pooled odds ratios (ORs) and the corresponding 95% confidence intervals (CIs). RESULTS: Finally, 14 eligible studies were identified, and alcohol consumption was not found to be a risk factor for rosacea. However, in subgroup analysis, alcohol consumption increased the risk of phymatous rosacea (PhR) and the pooled OR was 4.17 (95% CI = 1.76-9.91). CONCLUSION: Overall, our study showed that alcohol consumption was a risk factor in phymatous rosacea (PhR). More studies of rosacea investigating sex distribution, alcohol intake levels, and types of alcoholic beverages consumed are needed in the future.

18.
J Exp Clin Cancer Res ; 40(1): 297, 2021 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-34551797

RESUMO

BACKGROUND: Chemoresistance is the major cause of chemotherapy failure in patients with colorectal cancer (CRC). Protein tyrosine kinase 6 (PTK6) is aberrantly overexpressed in clinical CRC tissues undergoing chemotherapy. We studied if PTK6 contributed to the chemoresistance of CRC in human and mice. METHODS: We obtained tissue samples from patients with CRC and measured the expression of PTK6 by immunohistochemistry. Gain- and loss-of-function assays were performed to study the biological functions of PTK6. We constructed the FLAG-tagged wild type (WT), kinase-dead, and inhibition-defective recombinant mutants of PTK6 to study the effect phosphorylated activation of PTK6 played on CRC cell stemness and chemoresistance. We used small molecule inhibitor XMU-MP-2 to test the influence of PTK6 on sensitivity of CRC cells to 5-FU/L-OHP in both nude mouse and patient-derived xenograft (PDX) animal models. RESULTS: PTK6 is overexpressed in CRC tissues and plays a stimulatory role in the proliferation and chemoresistance of CRC cells both in vitro and in vivo. PTK6, especially the phosphorylated PTK6, can promote the stemness of CRC cells through interacting with JAK2 and phosphorylating it to activate the JAK2/STAT3 signaling. Pharmacological inhibition of PTK6 using XMU-MP-2 effectively reduces the stemness property of CRC cells and improves its chemosensitivity to 5-FU/L-OHP in both nude mice subcutaneously implanted tumor model and PDX model constructed with NOD-SCID mice. CONCLUSIONS: PTK6 interacts with JAK2 and phosphorylates it to activate JAK2/STAT3 signaling to promote the stemness and chemoresistance of CRC cells. Pharmacological inhibition of PTK6 by small molecule inhibitor dramatically enhances the sensitivity to chemotherapy in nude mice and PDX models.

19.
J Biochem ; 170(2): 195-202, 2021 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-34562091

RESUMO

The study intends to probe the functions of miR-142-5p in retinoblastoma (RB) and the relationship between miR-142-5p and phosphatase and tensin homolog deleted on chromosome ten (PTEN). In our study, miR-142-5p and PTEN mRNA expression in RB tissue, serum of RB patients and RB cell lines were investigated by quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation, migration, invasion and cell apoptosis were measured using MTT assay, BrdU assay, Transwell experiments and flow cytometry analysis, respectively. Binding sites between miR-142-5p and PTEN were predicted by the TargetScan database and were confirmed via qRT-PCR, western blot and dual-luciferase reporter gene assay. It was demonstrated that miR-142-5p expression was elevated in RB tissue, serum of RB patients and RB cell lines. MiR-142-5p overexpression remarkably promoted the proliferation, migration, invasion and inhibited the apoptosis of WERI-RB-1 cells while miR-142-5p knockdown induced opposite effects in Y79 cells. MiR-142-5p decreased PTEN expression in both mRNA and protein expression levels, and PTEN was identified as a target gene of miR-142-5p. Cotransfection of PTEN overexpression plasmids reversed the influences of miR-142-5p on RB cells. In conclusion, miR-142-5p enhances proliferation, migration and invasion of RB cell by targeting PTEN.


Assuntos
Movimento Celular/genética , Proliferação de Células/genética , MicroRNAs/genética , PTEN Fosfo-Hidrolase/genética , Neoplasias da Retina/genética , Retinoblastoma/genética , Apoptose/genética , Linhagem Celular Tumoral , Criança , Pré-Escolar , Biologia Computacional/métodos , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , MicroRNAs/metabolismo , Invasividade Neoplásica/genética , PTEN Fosfo-Hidrolase/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/métodos , Neoplasias da Retina/sangue , Neoplasias da Retina/metabolismo , Retinoblastoma/sangue , Retinoblastoma/metabolismo
20.
Microorganisms ; 9(7)2021 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-34361962

RESUMO

Shiga toxin-producing Escherichia coli (STEC) O103 strains have been recently attributed to various foodborne outbreaks in the United States. Due to the emergence of antibiotic-resistant strains, lytic phages are considered as alternative biocontrol agents. This study was to biologically and genomically characterize two STEC O103-infecting bacteriophages, vB_EcoP-Ro103C3lw (or Ro103C3lw) and vB_EcoM-Pr103Blw (or Pr103Blw), isolated from an organic farm. Based on genomic and morphological analyses, phages Ro103C3lw and Pr103Blw belonged to Autographiviridae and Myoviridae families, respectively. Ro103C3lw contained a 39,389-bp double-stranded DNA and encoded a unique tail fiber with depolymerase activity, resulting in huge plaques. Pr103Blw had an 88,421-bp double-stranded DNA with 26 predicted tRNAs associated with the enhancement of the phage fitness. Within each phage genome, no virulence, antibiotic-resistant, and lysogenic genes were detected. Additionally, Ro103C3lw had a short latent period (2 min) and a narrow host range, infecting only STEC O103 strains. By contrast, Pr103Blw had a large burst size (152 PFU/CFU) and a broad host range against STEC O103, O26, O111, O157:H7, and Salmonella Javiana strains. Furthermore, both phages showed strong antimicrobial activities against STEC O103:H2 strains. The findings provide valuable insight into these two phages' genomic features with the potential antimicrobial activities against STEC O103.

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