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1.
FASEB J ; 36 Suppl 12022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35555231

RESUMO

OBJECTIVE: Atrial septal defect (ASD) is one of the most common forms of congenital heart disease. Genetic defects play important roles in the pathogenesis of ASD but are complex and unclear. This study tries to discover the genetic pathogenesis in familial and sporadic ASD patients. METHODS: In Stage I, in a family with 30 members whole exome sequencing was performed in 4 ASD patients and the identified genetic variants were screened in the rest of the family members. In Stage II, 335 unrelated sporadic and isolated ASD patients were enrolled to test whether the genetic variants were involved in these patients. In Stage III, biological functions of the variants were elucidated at the cellular level and the CRISPR/Cas9 was used to construct variant-specific mutant mice in order to observe the abnormality of the heart structure. RESULTS: A novel heterozygous missense variant, FOXC1 c.518G>A (p.R173H) was identified in the ASD family by whole exome sequencing. Subsequently, other 4 heterozygous variants in FOXC1, including 2 novel missense variants: c.556-558delAAG (p.K186del) and c.559G>A (p.D187N) were identified in unrelated 335 sporadic ASD patients. In dual-luciferase reporter assay, the transcriptional activity of R173H decreased to almost 30% of the wild-type. In the Foxc1 R173H site-specific mutant mice, the atrial septum became very thin in tissue slices of the heart that was similar to the echocardiographic finding in the above family members. CONCLUSION: Variants in FOXC1 likely cause cardiac anomaly, particularly ASD. These findings provide a new insight into genetic mechanisms and counseling of familial and sporadic ASD.

2.
Front Pharmacol ; 13: 859951, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35559255

RESUMO

Retinal vein occlusion (RVO) is one of the most common retinal vascular diseases. The pathogenesis of RVO is multifactorial and involves a complex interplay among a variety of vascular and inflammatory mediators. Many cytokines, chemokines, growth factors, and cell adhesion molecules have been reported to be implicated. Treatments for RVO are directed at the management of underlying risk factors and vision-threatening complications, including macula edema (ME) and neovascularization. Intravitreal anti-VEGF agents are currently considered as the first-line treatment for ME secondary to RVO (RVO-ME), but a substantial proportion of patients responded insufficiently to anti-VEGF agents. Since RVO-ME refractory to anti-VEGF agents generally responds to corticosteroids and its visual outcome is negatively correlated to disease duration, prediction of treatment response at baseline in RVO-ME may significantly improve both cost-effectiveness and visual prognosis. Several bioactive molecules in the aqueous humor were found to be associated with disease status in RVO. This review aims to present a comprehensive review of intraocular biomolecules reported in RVO, including VEGF, IL-6, IL-8, MCP-1, sICAM-1, IL-12, IL-13, sVEGFR-1, sVEGFR-2, PDGF-AA, etc., highlighting their association with disease severity and/or phenotype, and their potential roles in prognostic prediction and treatment selection. Some of these molecules may serve as biomarkers for aqueous humor-based companion diagnostics for the treatment of RVO in the future.

3.
Theranostics ; 12(7): 3288-3315, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35547755

RESUMO

The advent of novel therapeutics in recent years has urged the need for a safe, non-immunogenic drug delivery vector capable of delivering therapeutic payloads specifically to diseased cells, thereby increasing therapeutic efficacy and reducing side effects. Extracellular vesicles (EVs) have garnered attention in recent years as a potentially ideal vector for drug delivery, taking into account their intrinsic ability to transfer bioactive cargo to recipient cells and their biocompatible nature. However, natural EVs are limited in their therapeutic potential and many challenges need to be overcome before engineered EVs satisfy the levels of efficiency, stability, safety and biocompatibility required for therapeutic use. Here, we demonstrate that an enzyme-mediated surface functionalization method in combination with streptavidin-mediated conjugation results in efficient surface functionalization of EVs. Surface functionalization using the above methods permits the stable and biocompatible conjugation of peptides, single domain antibodies and monoclonal antibodies at high copy number on the EV surface. Functionalized EVs demonstrated increased accumulation in target cells expressing common cancer associated markers such as CXCR4, EGFR and EpCAM both in vitro and in vivo. The functionality of this approach was further highlighted by the ability of targeting EVs to specifically deliver therapeutic antisense oligonucleotides to a metastatic breast tumor model, resulting in increased knockdown of a targeted oncogenic microRNA and improved metastasis suppression. The method was also used to equip EVs with a bifunctional peptide that targets EVs to leukemia cells and induces apoptosis, leading to leukemia suppression. Moreover, we conducted extensive testing to verify the biocompatibility, and safety of engineered EVs for therapeutic use, suggesting that surface modified EVs can be used for repeated dose treatment with no detectable adverse effects. This modular, biocompatible method of EV engineering offers a promising avenue for the targeted delivery of a range of therapeutics while addressing some of the safety concerns associated with EV-based drug delivery.

4.
Theranostics ; 12(7): 3057-3078, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35547766

RESUMO

Rationale: Spinal cord injury (SCI) remains an incurable neurological disorder leading to permanent and profound neurologic deficits and disabilities. Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are particularly appealing in SCI treatment to curtail damage, restore homeostasis and possible neural relay. However, the detailed mechanisms underlying hUC-MSC-mediated functional recovery of SCI have not been fully elucidated. The purpose of our current study is to identify novel therapeutic targets and depict the molecular mechanisms underlying the hUC-MSC-mediated recovery of subacute SCI. Methods: Adult female rats suffering from subacute incomplete thoracic SCI were treated with intrathecal transplantation of hUC-MSCs. The beneficial effects of hUC-MSCs on SCI repair were evaluated by a series of behavioral analyses, motor evoked potentials (MEPs) recording of hindlimb and immunohistochemistry. We carried out extensive transcriptome comparative analyses of spinal cord tissues at the lesion site from the subacute phase of SCI (sub-SCI) either treated without (+PBS) or with hUC-MSCs (+MSC) at 0 (sub-SCI), 1, 2, and 4 weeks post-transplantation (wpt), as well as normal spinal cord segments of intact/sham rats (Intact). Adeno-associated virus (AAV)-mediated neuron-specific expression system was employed to functionally screen specific γ-aminobutyric acid type A receptor (GABAAR) subunits promoting the functional recovery of SCI in vivo. The mature cortical axon scrape assay and transplantation of genetically modified MSCs with either overexpression or knockdown of brain-derived neurotrophic factor (BDNF) were employed to demonstrate that hUC-MSCs ameliorated the reduction of GABAAR subunits in the injured spinal cord via BDNF secretion in vitro and in vivo, respectively. Results: Comparative transcriptome analysis revealed the GABAergic synapse pathway is significantly enriched as a main target of hUC-MSC-activated genes in the injured spinal cord. Functional screening of the primary GABAAR subunits uncovered that Gabrb3 and Garbg2 harbored the motor and electrophysiological recovery-promoting competence. Moreover, targeting either of the two pivotal subunits ß3 or γ2 in combination with/without the K+/Cl- cotransporter 2 (KCC2) reinforced the therapeutic effects. Mechanistically, BDNF secreted by hUC-MSCs contributed to the upregulation of GABAAR subunits (ß3 & γ2) and KCC2 in the injured neurons. Conclusions: Our study identifies a novel mode for hUC-MSC-mediated locomotor recovery of SCI through synergistic upregulation of GABAAR ß3 and γ2 along with KCC2 by BDNF secretion, indicating the significance of restoring the excitation/inhibition balance in the injured neurons for the reestablishment of neuronal circuits. This study also provides a potential combinatorial approach by targeting the pivotal subunit ß3 or γ2 and KCC2, opening up possibilities for efficacious drug design.

5.
Molecules ; 27(9)2022 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-35566283

RESUMO

Procyanidins, as a kind of dietary flavonoid, have excellent pharmacological properties, such as antioxidant, antibacterial, anti-inflammatory and anti-tumor properties, and so they can be used to treat various diseases, including Alzheimer's disease, diabetes, rheumatoid arthritis, tumors, and obesity. Given the low bioavailability of procyanidins, great efforts have been made in drug delivery systems to address their limited use. Nowadays, the heavy burden of oral diseases such as dental caries, periodontitis, endodontic infections, etc., and their consequences on the patients' quality of life indicate a strong need for developing effective therapies. Recent years, plenty of efforts are being made to develop more effective treatments. Therefore, this review summarized the latest researches on versatile effects and enhanced bioavailability of procyanidins resulting from innovative drug delivery systems, particularly focused on its potential against oral diseases.

6.
BMC Med Educ ; 22(1): 356, 2022 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-35538519

RESUMO

BACKGROUND: A rigorous faculty appointment and promotion (FAP) system is vital for the success of any academic institution. However, studies examining the FAP system in Asian universities are lacking. We surveyed the FAP policies of Taiwan's medical schools and identified an overreliance on the CJA score (manuscript Category, Journal quality, and Author order). The potential shortcomings of this metric and recommendations for refinement were discussed. METHODS: We obtained the FAP documents from all 12 medical schools in Taiwan, and analyzed their use of traditional versus non-traditional criteria for FAP according to a published methodology. The influence of the journal impact factor (JIF) on the FAP process was quantified by comparing its relative weight between papers with two extreme JIFs. To better understand the research impact and international standing of each school, we utilized the public bibliographic database to rank universities by the number of papers, and the proportions of papers within the top 10% or 50% citation. RESULTS: Compared with other countries, Taiwan's medical schools focus more on the quantifiable quality of the research, mostly using a "CJA" score that integrates the category, JIF or ranking, and authorship of a paper, with the JIF being the most influential factor. The CJA score for an article with a JIF of 20 can be up to three times the threshold for promotion to Assistant Professor. The emphasis on JIF is based on a presumed correlation between JIF and citation counts. However, our analysis shows that Taiwan's medical schools have lower-than-average citation counts despite a competitive rank in the number of publications. CONCLUSIONS: The JIF plays an unrivaled role in determining the outcome of FAP in Taiwan's medical schools, mostly via the CJA system. The questionable effectiveness of the current system in elevating the international standing of Taiwan's higher-education institutions calls for a re-examination of the FAP system. We recommend a reduction in the relative importance of CJA score in the FAP system, adopting more rigorous metrics such as the h-index for evaluating research quality, and supporting more research aimed at improving the FAP system.


Assuntos
Fator de Impacto de Revistas , Faculdades de Medicina , Autoria , Docentes , Docentes de Medicina , Humanos , Taiwan
7.
Int J Oral Sci ; 14(1): 25, 2022 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-35538067

RESUMO

Head and neck squamous cell carcinoma (HNSCC) still lacks effective targeted treatment. Therefore, exploring novel and robust molecular targets is critical for improving the clinical outcome of HNSCC. Here, we reported that the expression levels of family with sequence similarity 64, member A (FAM64A) were significantly higher in HNSCC tissues and cell lines. In addition, FAM64A overexpression was found to be strongly associated with an unfavorable prognosis of HNSCC. Both in vitro and in vivo evidence showed that FAM64A depletion suppressed the malignant activities of HNSCC cells, and vice versa. Moreover, we found that the FAM64A level was progressively increased from normal to dysplastic to cancerous tissues in a carcinogenic 4-nitroquinoline-1-oxide mouse model. Mechanistically, a physical interaction was found between FAM64A and forkhead box protein M1 (FOXM1) in HNSCC cells. FAM64A promoted HNSCC tumorigenesis not only by enhancing the transcriptional activity of FOXM1, but also, more importantly, by modulating FOXM1 expression via the autoregulation loop. Furthermore, a positive correlation between FAM64A and FOXM1 was found in multiple independent cohorts. Taken together, our findings reveal a previously unknown mechanism behind the activation of FOXM1 in HNSCC, and FAM64A might be a promising molecular therapeutic target for treating HNSCC.


Assuntos
Neoplasias de Cabeça e Pescoço , Animais , Carcinogênese , Linhagem Celular Tumoral , Proliferação de Células , Transformação Celular Neoplásica , Neoplasias de Cabeça e Pescoço/genética , Homeostase , Camundongos , Carcinoma de Células Escamosas de Cabeça e Pescoço
8.
J Hazard Mater ; 436: 129123, 2022 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-35596988

RESUMO

The efficient deployment of visible and near-infrared (NIR) light for photocatalytic disinfection is of great concern a matter. Herein, we report a specific bifunctional 2D/2D g-C3N4/BiO2-x nanosheets heterojunction, prepared through a self-assembly approach. Delightfully, the obtained 2D/2D heterojunctions exhibited satisfactory photocatalytic disinfection performance towards Escherichia coli K-12 (E. coli K-12) under visible light irradiation, which was credited to the Z-scheme interfacial heterojunction facilitating the migration of photogenerated carries. The photoactivity enhancement driven by NIR light illumination was ascribed to the cooperative synergy effect of photothermal effect and "hot electrons", engineering efficient charge transfer. Intriguingly, the carboxyl groups emerged on g-C3N4 nanosheets contributed a vital role in establishing the enhanced photocatalytic reaction. Moreover, the disinfection mechanism was systematically described. The cell membrane was destroyed, evidenced by the generation of lipid peroxidation reaction and loss of energy metabolism. Subsequently, the damage of defense enzymes and release of intracellular constituents announced the irreversible death of E. coli K-12. Interestingly enough, considerable microbial community shifts of surface water were observed after visible and NIR light exposure, highlighting the critical feature of disinfection process in shaping microbial communities. The authors believe that this work gives a fresh light on the feasibility of heterostructures-enabled disinfection processes.

9.
J Oncol ; 2022: 6449984, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35607324

RESUMO

Background: Triterpenoid saponins from sea cucumbers exhibit significant antitumour, antifungal, and antibacterial activities. However, the associated molecular mechanisms have yet to be elucidated. In this study, we screened and explored the antitumour activity and underlying mechanisms of triterpenoid saponins isolated from Thelenota ananas. Methods: We isolated and purified sea cucumber saponins, determined their chemical structures, and confirmed their function in vitro. We also screened and explored the antitumour activity and underlying mechanisms of triterpenoid saponins isolated from Thelenota ananas. Results: Four saponins were discovered from sea cucumber Thelenota ananas collected from the South China Sea. We found that stichloroside C2 (STC2) inhibited the proliferation and clonogenesis of the human triple-negative breast cancer (TNBC) cell line MDA-MB-231 and mouse TNBC cell line 4 T1 in a dose-dependent manner and induced apoptosis and cycle arrest in these two TNBC cell lines. STC2 induced DNA damage in two TNBC cell lines and significantly increased the protein expression level of the DNA double-strand break marker γ-H2AX. STC2 downregulated the protein expression levels of phosphorylated cyclin-dependent kinase 1 (CDK1), cyclin B1, CDK2, and cyclin A2 in MDA-MB-231 and 4 T1 cells. STC2 upregulated Bax and cleaved PARP protein expression in two types of breast cancer cells. In addition, STC2 promoted E-cadherin expression; inhibited vimentin expression; upregulated the phosphorylation levels of the mitogen-activated protein kinase (MAPK) signalling pathway-related proteins p38, JNK, and ERK1/2; and downregulated Akt phosphorylation. Conclusions: STC2 exerts anti-TNBC activity, inhibits epithelial-mesenchymal transition (EMT), and induces apoptosis by regulating the cell cycle, EMT-related proteins, and MAPK signalling pathway.

10.
Endocr Relat Cancer ; 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35521773

RESUMO

Adrenocortical carcinoma (ACC) is a rare malignancy with poor prognosis and challenging management. The present study aimed to investigate expression of programmed death ligand-1 (PD-L1) and V-domain Ig-containing suppressor of T cell activation (VISTA) in ACC and their associations with clinicopathological features and survival outcomes. Immunohistochemistry was performed on formalin-fixed paraffin-embedded specimens from 54 ACC patients. Chi-square/Fisher's exact test or independent samples t/U-Mann-Whitney tests were applied to assess correlations between immunoscores and clinicopathological parameters. The Kaplan-Meier method and Cox regression were conducted for survival analysis and to identify independent predictors of overall (OS) and disease-free (DFS) survival. Results showed that VISTA was expressed in tumor cells (TCs) and tumor-infiltrating immune cells (TICs) in 81.5% (44/54) and 40.7% (22/54) of the patients, respectively. PD-L1 positivity was found in either TCs or TICs in 11.1% (6/54) of the patients. Patients with positive VISTA expression in TCs exhibited a higher tumor stage (56.9% vs. 20%, p = 0.036) and Ki67 index (30.50 ± 23.51% vs. 14.76 ± 11.75%, p = 0.006). However, PD-L1 positivity in either TCs or TICs had no association with patient clinicopathological features. Higher VISTA expression intensity, area and immunoscore were associated with increased risks of disease progression and overall mortality, but PD-L1 expression in TCs or TICs did not contribute to OS or DFS. In conclusion, positive TC VISTA expression correlates with pathological parameters related to malignancy in ACC. This finding provides novel evidence for VISTA to serve as an immunotherapeutic target in addition to PD-L1 for ACC.

11.
Toxicol Appl Pharmacol ; 446: 116045, 2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35526792

RESUMO

Xylitol is a hygroscopic compound known to protect nasal cavity against bacteria. It has also been developed into nasal spray and evaluated as a potential candidate drug for respiratory diseases. Consequently, it is necessary to study its inhalation toxicity. Based on our previous study on its subacute inhalation toxicity, this study aimed to investigate the safety of xylitol inhalation for long-term use. According to the OECD Test Guideline 413, Sprague-Dawley rats were randomly divided into six groups and exposed with different concentrations of xylitol aerosol or air. After exposure for 90-day, the recovery groups were continued to observe for a recovery period of 28-day. No significant changes in body weight were observed between sham and xylitol groups. Several significant differences in hematological, clinical chemistry, bronchoalveolar lavage fluid were observed, which either had no dose-effect relationship for both male and female rats or were restored during the recovery period. Finally, except for high dose group of xylitol, two rats showed a small amount of inflammatory exudate in alveolar and bronchial cavities, which was restored in the recovery period. The rest of rats showed no obvious difference. For the recovery groups, no significant difference was observed between these two groups. In conclusion, the no observable adverse effect level (NOAEL) of xylitol in our subchronic inhalation toxicological experiments was 2.9 mg/L, which indicated that xylitol for rats' long-time inhalation is tolerant and safe.

12.
Oxid Med Cell Longev ; 2022: 7412208, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35528520

RESUMO

Purpose: Diabetic macular edema (DME) is a major cause of vision loss in patients with diabetic retinopathy; this study is aimed at comparing the expression of aquaporins (AQPs) on the inner limiting membranes (ILMs) of various vitreoretinal diseases and investigating the role of aquaporins expressed on the ILMs in mediating the occurrence of DME. Methods: The whole-mounted ILM specimens surgically excised from patients with various vitreoretinal diseases (idiopathic macular hole, myopic traction maculopathy, and diabetic retinopathy) were analyzed by immunohistochemistry (IHC). The distribution and morphology of AQP4, AQP7, and AQP11 on the ILMs were correlated with immunohistochemical staining characteristics. Moreover, immunofluorescence of AQP4 was performed on the ILM specimens of the patient in four groups: the control group, negative control group, no DME group, and DME group. The immunofluorescence intensity value of AQP4 was measured using ImageJ. The difference between the four groups and the correction between the immunofluorescence value and central foveal thickness (CFT) were analyzed. Results: In IHC sections, the expression of AQP4, AQP7, and AQP11 on ILMs of diabetic retinopathy (DR) with macular edema, respectively, seemed to be more abundant than in the idiopathic macular hole (iMH) and myopic traction maculopathy (MTM). Moreover, markedly higher fluorescence intensity of AQP4 of ILMs was determined in the DME group (51.05 ± 5.67) versus the other three groups (P < 0.001). A marked positive association was identified between the fluorescence intensity of AQP4 and CFT (r = 0.758; P = 0.011). Conclusions: AQP4, AQP7, and AQP11 can be expressed on human ILM in vivo. The increased expression of AQPs on the ILMs of DR may be associated with the occurrence of DME. Moreover, the degree of DME may be positively correlated with the expression of AQP4 on the ILMs.


Assuntos
Aquaporinas , Diabetes Mellitus , Retinopatia Diabética , Edema Macular , Doenças Retinianas , Perfurações Retinianas , Aquaporinas/metabolismo , Membrana Basal/metabolismo , Diabetes Mellitus/metabolismo , Retinopatia Diabética/metabolismo , Humanos , Doenças Retinianas/metabolismo , Perfurações Retinianas/complicações , Perfurações Retinianas/metabolismo , Vitrectomia
13.
Front Cell Neurosci ; 16: 872347, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35530178

RESUMO

Dysregulated synaptic plasticity is a key feature of neurodevelopmental disorders, including autism. This study investigated whether Fragile X mental retardation protein (FMRP), a selective RNA-binding protein that regulates synaptic protein expression by interacting with miRNAs, mediates the effects of androgens that play an important role in regulating the synaptic plasticity in the hippocampus. Experiments using mouse hippocampal neuron HT22 cells demonstrated that dihydrotestosterone (DHT) increased the expression of postsynaptic density protein 95 (PSD95) by inhibiting FMRP expression. Administration of miR-125a inhibitor upregulated the PSD95 expression and significantly increased the DHT-induced upregulation of PSD95. FMRP knockdown in HT22 cells reduced the expression of miR-125a. Moreover, miR-125a inhibitor upregulated the PSD95 expression in the DHT-treated HT22 cells with FMRP knockdown. Subsequently, the effects of androgen-mediated via FMRP in regulating neural behaviors and PSD95 expression and dendritic spines density/morphology were investigated using Fmr1 knockout (KO) and wild-type littermate (WT) mice. The castration of WT mice reduced the androgen levels, aggravated anxiety and depression, and impaired learning and memory and sociability of mice. DHT supplementation post-castration reversed the alterations in density and maturity of dendritic spines of hippocampal neurons and behavioral disorders in WT mice; however, it did not reveal such effects in Fmr1 KO mice. Further, immunohistochemical staining and western blotting analyses after knocking down miR-125a revealed similar effects of castration and post-castration DHT supplementation on PSD95 protein expression. These findings clarified that FMRP mediated the effects of DHT through miR-125a in regulating the expression of hippocampal synaptic protein PSD95. This study provides evidence for the neuroprotective mechanism of androgen in PSD95 expression and dendritic spines density/morphology and suggests that treatment interventions with androgen could be helpful for the management of synaptic plasticity disorders.

14.
FASEB J ; 36 Suppl 12022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35560658

RESUMO

OBJECTIVES: Congenital heart disease (CHD) is a common disease of human birth defects, and atrial septal defect (ASD) is the most common form of ASD. Genetic variants in the coding region of the CITED2 gene is known to be significantly correlated with cardiac malformations, but variants of the CITED2 promoter region and its relationship with the formation of ASD are still unclear. We hypothesize that the variants of the CITED2 promoter may be related to pathogenesis of ASD. The purpose of this study was to screen variants in the promoter region of the CITED2 gene and to verify the effect of the variants on gene expression at the cellular level. METHODS: The blood samples of 332 ASD patients and 293 unrelated healthy children used as controls were studied. The total DNA of all subjects was extracted, and the CITED2 promoter variants were screened by PCR combined with Sanger sequencing. The luciferase activity of the CITED2 promoter was measured by a dual luciferase reporter at the cellular level. Electrophoretic mobility change assay (EMSA) and bioinformatics analysis were used to test the effect of CITED2 promoter changes on transcription factor binding sites. RESULTS: Four variants in the promoter region of the CITED2 gene were found only in the 332 ASD patients with zero occurrence in the 293 control subjects. These included one novel heterozygous variant (g.4933C>A) and three SNPs [1 case of g.4078 A>C(rs1165649373), 4 cases of g.4935C>T(rs111470468), 2 cases of g.5027C>T (rs112831934)]. The cellular functional analysis showed that these four variants significantly changed the transcriptional activity of the CITED2 gene promoter in HEK-293 cells (P<0.05). In addition, EMSA results and database analysis showed that these variants created or destroyed a series of possible transcription factor binding sites, resulting in changes of the expression of CITED2 protein. CONCLUSION: The variants of CITED2 promoter sequence in ASD patients affect transcriptional activity and are likely involved in the occurrence and development of ASD. These findings may provide new perspectives on the molecular pathogenesis and potential therapeutic insights of ASD patients.

15.
FASEB J ; 36 Suppl 12022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35560747

RESUMO

OBJECTIVES: Post-translational modifications (PTMs) can modify proteins and regulate their functions. Lysine crotonylation is a novel protein PTM that can be regulated by silent mating type information regulation 2 homolog 1 (SIRT1). Ca2+ /calmodulin-dependent protein kinase II (CaMKII) plays a significant role in mediating cellular responses to external signaling molecules. Little is known regarding the effect of SIRT1 on CaMKII crotonylation and the impact of crotonylation on the activity of this enzyme. METHODS: Cre-Loxp technology was used to generate conditional cardiac-specific SIRT1 knockout mice (SIRT1 KO). Protein crotonylation modification was studied by PTMomics. CaMKII activity was determined by ELISA. Western blot and co-immunoprecipitation (CO-IP) were performed to examine the expression and crotonylation of CaMKII RESULTS: PTMomics showed changes in protein crotonylation profiles in the myocardium of cardiac-specific SIRT1 knockout mice. Crotonylation of CaMKII was significantly increased at lysine 301 and the enhanced CaMKII crotonylation was further confirmed by CO-IP (2.37±0.25 in SIRT1 KO vs. 0.96±0.10 in wild-type and 1.04±0.10 in SIRT1fl/fl mice, p<0.01). Cardiac-specific knockout of SIRT1 did not affect the protein expression of CaMKII whereas significantly increased CaMKII activity (2.75±0.17 m units/µL in SIRT1 KO vs. 2.10±0.13 m units/µL in wild-type and 1.98±0.17 m units/µL in SIRT1fl/fl mice, p<0.05). CO-IP showed no physical interaction between SIRT1 and CaMKII. CONCLUSIONS: SIRT1 deficiency increases CaMKII activity in myocardium at least partially through enhancing lysine crotonylation of CaMKII, which advances our understanding of the role played by PTM in the regulation of CaMKII.

16.
FASEB J ; 36 Suppl 12022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35560955

RESUMO

OBJECTIVE: Atrial septal defect (ASD) is one of the most common types of congenital heart disease (CHD). As a transcription factor, ISL1 is expressed specifically in second heart field during embryogenesis and plays a crucial role in heart development and cardiovascular morphogenesis. Studies have shown association of ISL1 multiple variants with diverse type of CHD including ASD, whereas the variants of ISL1 gene promoter region in ASD patients have not been reported. We hypothesized that variants in ISL1 promoter may be involved in the formation of ASD. METHODS: Blood samples from 593 subjects, including 300 patients with isolated ASD and 293 healthy controls were studied. Total DNA from all subjects was extracted and the PCR combined with Sanger sequencing was used to identify ISL1 gene promoter variants. Functional analysis of DNA sequence variants was performed using a dual luciferase reporter assay. Electrophoretic mobility shift assay (EMSA) was selected to examine the influence of the variants in ISL1 gene promoter on binding of transcription factors. All possible binding sites of transcription factor influenced by the identified variants were predicted by the PROMO database. RESULTS: Four variants in ISL1 gene promoter were found only in patients with ASD by sequencing. Three of the 4 variations [g.4923 G>C (rs541081886), g.5079 A>G (rs1371835943), g.5309 G>A (rs116222082)] significantly decreased the transcriptional activities compared with the wild-type ISL1 gene promoter (P < 0.05). The EMSA revealed that these variants [g.4923 G>C (rs541081886), g.5079 A>G (rs1371835943), g.5309 G>A (rs116222082)] in ISL1 promoter affected the number and affinity of binding sites of transcription factors. Further analysis with the online PROMO database demonstrated that a cluster of putative binding sites for transcription factors may be altered by these variants. CONCLUSION: These sequence variants identified from the promoter region of ISL1 gene in ASD patients are likely involved in the development of ASD by affecting the transcriptional activity and altering ISL1 levels. Therefore, these findings may provide new insights into the molecular etiology and potential therapeutic strategy of ASD.

17.
Cell Mol Biol Lett ; 27(1): 36, 2022 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-35562668

RESUMO

BACKGROUND: RING finger protein 7 (RNF7) is a highly conserved protein that functions as an E3 ubiquitin ligase. RNF7 overexpression is indicated in multiple human cancers, but its role in renal cell carcinoma (RCC) and the mechanisms underlying how it regulates the initiation and progression of RCC have not been explored. METHODS: Bioinformatics analysis, quantitative reverse-transcription polymerase chain reaction (RT-PCR), and Western blot were conducted to determine the expression of RNF7 in RCC tissues and cell lines. Knockdown and overexpression experiments were performed to examine the effects of RNF7 on cell viability, apoptosis, and glycolysis in vitro and on tumor growth in nude mice in vivo. RESULTS: The elevated RNF7 expression in tumor tissues of patients with RCC was correlated with poor survival. RNF7 overexpression inhibited apoptosis and promoted glycolysis in vitro and increased tumor growth in vivo by activating the JAK/STAT3 signaling pathway by ubiquitination of SOCS1. Moreover, RNF7 overexpression affected the sensitivity of RCC cells to sunitinib. Finally, STAT3 activation was necessary for transcriptional induction of RNF7. CONCLUSION: These results demonstrate that RNF7 inhibited apoptosis, promoted glycolysis, and inhibited sunitinib sensitivity in RCC cells via ubiquitination of SOCS1, thus activating STAT3 signaling. These suggest the potential for targeting the RNF7-SOCS1/JAK/STAT3 pathway for RCC treatment.

18.
Front Med (Lausanne) ; 9: 815450, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35510248

RESUMO

Globally, cervical cancer (CC) is the most common malignant tumor of the female reproductive system and its incidence is only second after breast cancer. Although screening and advanced treatment strategies have improved the rates of survival, some patients with CC still die due to metastasis and drug resistance. It is considered that cancer is driven by somatic mutations, such as single nucleotide, small insertions/deletions, copy number, and structural variations, as well as epigenetic changes. Previous studies have shown that cervical intraepithelial neoplasia is associated with copy number variants (CNVs) and/or mutations in cancer-related genes. Further, CC is also related to genetic mutations. The present study analyzed the data on somatic mutations of cervical squamous cell carcinoma (CESC) in the Cancer Genome Atlas database. It was evident that the Apolipoprotein B mRNA editing enzyme-catalyzed polypeptide-like (APOBEC)-related mutation of the FLG gene can upregulate the expression of the JUN gene and ultimately lead to poor prognosis for patients with CC. Therefore, the findings of the current study provide a new direction for future treatment of CC.

19.
Exp Hematol Oncol ; 11(1): 25, 2022 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-35505384

RESUMO

BACKGROUND: Acute graft-versus-host disease (aGVHD) remains the major cause of early mortality after haploidentical related donor (HID) hematopoietic stem cell transplantation (HSCT). We aimed to establish a comprehensive model which could predict severe aGVHD after HID HSCT. METHODS: Consecutive 470 acute leukemia patients receiving HID HSCT according to the protocol registered at https://clinicaltrials.gov (NCT03756675) were enrolled, 70% of them (n = 335) were randomly selected as training cohort and the remains 30% (n = 135) were used as validation cohort. RESULTS: The equation was as follows: Probability (grade III-IV aGVHD) = [Formula: see text], where Y = -0.0288 × (age) + 0.7965 × (gender) + 0.8371 × (CD3 + /CD14 + cells ratio in graft) + 0.5829 × (donor/recipient relation) - 0.0089 × (CD8 + cell counts in graft) - 2.9046. The threshold of probability was 0.057392 which helped separate patients into high- and low-risk groups. The 100-day cumulative incidence of grade III-IV aGVHD in the low- and high-risk groups was 4.1% (95% CI 1.9-6.3%) versus 12.8% (95% CI 7.4-18.2%) (P = 0.001), 3.2% (95% CI 1.2-5.1%) versus 10.6% (95% CI 4.7-16.5%) (P = 0.006), and 6.1% (95% CI 1.3-10.9%) versus 19.4% (95% CI 6.3-32.5%) (P = 0.017), respectively, in total, training, and validation cohort. The rates of grade III-IV skin and gut aGVHD in high-risk group were both significantly higher than those of low-risk group. This model could also predict grade II-IV and grade I-IV aGVHD. CONCLUSIONS: We established a model which could predict the development of severe aGVHD in HID HSCT recipients.

20.
Int J Biol Macromol ; 209(Pt A): 315-329, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35405151

RESUMO

In this study, we used a combination of chemical and biological pretreatment methods to extract cellulose from corn straw with a relative content of 92.40%. The adsorption performance and mechanism of the prepared porous carbon were investigated using synthetic dye malachite green (MG) and antibiotic tetracycline hydrochloride (TC) as adsorption models. The kinetic studies suggested that the adsorption followed the pseudo-second-order model and Bangham model. The Freundlich isotherm model fitted the adsorption data best for both MG and TC. The thermodynamic studies showed that the adsorption of MG and TC by adsorbents were spontaneous and endothermic in nature. In addition, the adsorption performance was maintained at 50% of the original value after five cycles. More importantly, this method not only improved the adsorption performance of prepared porous carbon materials but also provides a reference for the application of other lignocellulosic materials for cellulose extraction.

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