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1.
Ann Transl Med ; 10(16): 860, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36110992

RESUMO

Background: Glioma is the most common primary intracranial tumor with poor prognosis. The prediction of glioma prognosis has not been well investigated. XGBoost algorithm has been widely used in and data analysis. The predictive value of XGBoost algorithm in glioma remains unclear. This current study used the XGBoost algorithm to construct a predictive model for postoperative outcomes of glioma patients. Methods: Patients with glioma who underwent surgery from January 2006 to April 2017 were retrospectively included in this study. Clinical and follow-up data were collected. The XGBoost model and multivariate logistic regression analysis model were used to screen the factors related to postoperative outcomes, and the results of the two models were compared. The area under the receiver operating characteristic (ROC) curve (AUC), sensitivity, specificity, and Youden index were calculated to evaluate the predictive value of the XGBoost model. Results: A total of 638 patients were included. In total, 336 (52.7%) cases died within 5 years after the operation. Multivariate logistic regression analysis showed that age, gender, World Health Organization (WHO) grade, extent of tumor resection, Karnofsy performance score (KPS), tumor diameter, and whether postoperative radiotherapy and chemotherapy were administered, were the most important risk factors for death within 5 years after surgery in glioma patients. The XGBoost model showed that the top 5 factors related to death of glioma patients within 5 years after surgery were WHO grade (30 points), extent of tumor resection (19 points), postoperative radiotherapy and chemotherapy (16 points), KPS (14 points), and age (11 points). The AUC of the XGBoost model for predicting the death of glioma patients within 5 years after surgery was 0.803 [95% confidence interval (CI): 0.718-0.832], and the sensitivity and specificity were 0.894 and 0.581, respectively. The Youden index was 0.475. The AUC of the multivariate logistic regression model was 0.738 (95% CI: 0.704-0.781), the sensitivity and specificity were 0.785 and 0.632, respectively, and the Youden index was 0.417. Conclusions: Compared with multivariate logistic regression model, XGBoost model has better performance in predicting the risk of death within 5 years after surgery in patients with glioma.

2.
Clin Transl Med ; 12(9): e990, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36103411

RESUMO

BACKGROUND: The number of women delivering at advanced maternal age (AMA; > = 35) continuously increases in developed and high-income countries. Large cohort studies have associated AMA with increased risks of various pregnancy complications and adverse pregnancy outcomes, which raises great concerns about the adverse effect of AMA on the long-term health of offspring. Specific acquired characteristics of parents can be passed on to descendants through certain molecular mechanisms, yet the underlying connection between AMA-related alterations in parents and that in offspring remains largely uncharted. METHODS: We profiled the DNA methylomes of paired parental peripheral bloods and cord bloods from 20 nuclear families, including 10 AMA and 10 Young, and additional transcriptomes of 10 paired maternal peripheral bloods and cord bloods. RESULTS: We revealed that AMA induced aging-like changes in DNA methylome and gene expression in both parents and offspring. The expression changes in several genes, such as SLC28A3, were highly relevant to the disorder in DNA methylation. In addition, AMA-related differentially methylated regions (DMRs) identified in mother and offspring groups showed remarkable similarities in both genomic locations and biological functions, mainly involving neuron differentiation, metabolism, and histone modification pathways. AMA-related differentially expressed genes (DEGs) shared by mother and offspring groups were highly enriched in the processes of immune cell activation and mitotic nuclear division. We further uncovered developmental-dependent dynamics for the DNA methylation of intergenerationally correlated DMRs during pre-implantation embryonic development, as well as diverse gene expression patterns during gametogenesis and early embryonic development for those common AMA-related DEGs presenting intergenerational correlation, such as CD24. Moreover, some intergenerational DEGs, typified by HTRA3, also showed the same significant alterations in AMA MII oocyte or blastocyst. CONCLUSIONS: Our results reveal potential intergenerational inheritance of both AMA-related DNA methylome and transcriptome and provide new insights to understand health problems in AMA offspring.


Assuntos
Epigenoma , Transcriptoma , Metilação de DNA/genética , Epigenoma/genética , Feminino , Humanos , Padrões de Herança , Idade Materna , Gravidez , Serina Endopeptidases/genética , Transcriptoma/genética
3.
J Control Release ; 350: 298-307, 2022 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-36002054

RESUMO

Chimeric receptor T cells (CAR-T) can effectively cure leukemia; however, there are two limitations: a complicated preparation process ex vivo and cytokine release syndrome (CRS). In this study, we constructed a lipid nanoparticle system modified by CD3 antibody on the surface, loading with the plasmid containing the combination gene of interleukin 6 short hairpin RNA (IL-6 shRNA) and CD19-CAR (AntiCD3-LNP/CAR19 + shIL6). The system targeted T cells by the mediation of CD3 antibody and stably transfected T cells to transform them into CAR-T cells with IL-6 knockdown, thus killing CD19-highly expressed leukemia tumor cells and reducing CRS caused by IL-6. In vivo experiments showed that AntiCD3-LNP/CAR19 + shIL6 could stably transfect T cells and produce CAR-T within 90 days to kill the tumor. This significantly prolonged the survival time of leukemia model mice and demonstrated the prepared LNP exhibited the same anti-tumor effect as the traditional CAR-T cells prepared ex vivo. In this study, CAR-T cells were directly produced in vivo after intravenous injection of the lipid nanoparticles, without the need of using the current complex process ex vivo. Additionally, IL-6 expression was silenced, which would be helpful to reduce the CRS and improve the safety of CAR-T therapy. This method improves the convenience of using CAR-T technology and is helpful in further promoting the clinical application of CAR-T.

4.
PLoS Genet ; 18(8): e1010310, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35939513

RESUMO

Chromosomal mosaicism is common throughout human pre- and post-implantation development. However, the incidence and characteristics of mosaicism in human blastocyst remain unclear. Concerns and confusions still exist regarding the interpretation of chromosomal mosaicism on preimplantation genetic testing for aneuploidy (PGT-A) results and embryo development. Here, we aimed to estimate the genetic concordance between trophectoderm (TE), inner cell mass (ICM) and the corresponding human embryonic stem cells (hESCs), and to explore the characteristics of mosaicism in human blastocyst and hESCs on a single cell level. The single cell sequencing results of TE cells indicated that 65.71% of the blastocysts were mosaic (23 in 35 embryos), while the ICM sequencing results suggested that 60.00% of the blastocysts were mosaic (9 in 15 embryos). The incidence of mosaicism for the corresponding hESCs was 33.33% (2 in 6 embryos). No significant difference was observed between the mosaic rate of TE and that of ICM. However, the mosaic rate of the corresponding hESCs was significantly lower than that of TE and ICM cells, suggesting that the incidence of mosaicism may decline during embryonic development. Upon single cell sequencing, we found several "complementary" copy number variations (CNVs) that were usually not revealed in clinical PGT-A which used multi-cell DNA sequencing (or array analysis). This indicates the potential diagnostic risk of PGT-A based multi-cell analysis routinely in clinical practice. This study provided new insights into the characteristics, and considerable influences, of mosaicism on human embryo development, as well as the clinical risks of PGT-A based on multi-cell biopsies and bulk DNA assays.


Assuntos
Mosaicismo , Diagnóstico Pré-Implantação , Aneuploidia , Blastocisto , Variações do Número de Cópias de DNA/genética , Feminino , Testes Genéticos/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Gravidez , Diagnóstico Pré-Implantação/métodos
5.
Sci Adv ; 8(32): eabj3725, 2022 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-35947672

RESUMO

Mechanisms of implantation such as determination of the attachment pole, fetal-maternal communication, and underlying causes of implantation failure are largely unexplored. Here, we performed single-cell RNA sequencing on peri-implantation embryos from both humans and mice to explore trophectoderm (TE) development and embryo-endometrium cross-talk. We found that the transcriptomes of polar and mural TE diverged after embryos hatched from the zona pellucida in both species, with polar TE being more mature than mural TE. The implantation poles show similarities in cell cycle activities, as well as in expression of genes critical for implantation and placentation. Embryos that either fail to attach in vitro or fail to implant in vivo show abnormalities in pathways related to energy production, protein metabolism, and 18S ribosomal RNA m6A methylation. These findings uncover the gene expression characteristics of humans and mice TE differentiation during the peri-implantation period and provide new insights into embryo implantation.

6.
Reprod Biomed Online ; 45(3): 473-480, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35780041

RESUMO

RESEARCH QUESTION: Can preimplantation genetic testing for structural rearrangement (PGT-SR) based on low-coverage next-generation sequencing (NGS) accurately discriminate between normal and carrier embryos of reciprocal translocation (RecT) and Robertsonian translocation (RobT)? DESIGN: A total of 109 couples with RecT or RobT were included in this study. The ages, bad obsteric histories (BOH), blood karyotype and IVF cycle information, including the number of cumulus-oocyte complexes, metaphase II oocytes, two pronuclei oocytes and blastocysts were recorded. 0.1 × whole genome sequencing (WGS) of embryos followed by copy number variation (identifying unbalanced/balanced) and 2 × WGS of parents and embryos followed by haplotype analysis (discriminating between normal and carrier) were carried out in PGT-SR cycles. The embryos without translocation were transferred and clinical outcomes evaluated. RESULTS: Among all the couples in this study, 67 patients had RecT and 42 had RobT. After unbalanced and balanced detection, 103 balanced embryos underwent a further normal and carrier discrimination procedure, and 53 normal embryos were identified. Finally, 32 normal embryos were transferred, with an ongoing pregnancy rate of 46.88% (15/32). All ongoing pregnancies underwent amniocentesis, and the amninocentesis karyotyping results showed 100% concordance with PGT-SR diagnosis. CONCLUSIONS: Our low-coverage NGS-based PGT-SR method can accurately discriminate between normal and carrier status of balanced embryos. The method is cost-effective and has broad clinical applicability.


Assuntos
Transferência Embrionária , Diagnóstico Pré-Implantação , Aberrações Cromossômicas , Variações do Número de Cópias de DNA , Transferência Embrionária/métodos , Feminino , Fertilização In Vitro , Testes Genéticos/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Gravidez , Diagnóstico Pré-Implantação/métodos , Translocação Genética
7.
Nat Commun ; 13(1): 3814, 2022 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-35780140

RESUMO

Birds, reptiles and insects have the ability to discriminate humidity levels that influence their survival and geographic distribution. Insects are particularly susceptible to humidity changes due to high surface area to volume ratios, but it remains unclear how humidity sensors transduce humidity signals. Here we identified Or42b-expressing olfactory sensory neurons, which are required for moisture attraction in Drosophila. The sensilla housing Or42b neurons show cuticular deformations upon moist air stimuli, indicating a conversion of humidity into mechanical force. Accordingly, we found Or42b neurons directly respond to humidity changes and rely on the mechanosensitive ion channel TMEM63 to mediate humidity sensing (hygrosensation). Expressing human TMEM63B in Tmem63 mutant flies rescued their defective phenotype in moisture attraction, demonstrating functional conservation. Thus, our results reveal a role of Tmem63 in hygrosensation and support the strategy to detect humidity by transforming it into a mechanical stimulus, which is unique in sensory transduction.


Assuntos
Proteínas de Drosophila , Neurônios Receptores Olfatórios , Animais , Drosophila/genética , Proteínas de Drosophila/genética , Umidade , Insetos , Sensilas
8.
3 Biotech ; 12(9): 188, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35879964

RESUMO

Floret opening and closure are critical for rice to complete reproductive development. To further understand the molecular mechanism of floret opening and closure in rice, RNA-seq was performed on the floret of Indica and Japonica rice in the state of 1-h before floret opening, at the opening and closure, respectively. Our results show that many differentially expressed genes are produced throughout the floret opening and closure of both Indica and Japonica rice. Differentially expressed genes shared between Indica and Japonica rice at floret opening were involved in seven metabolic pathways, including plant hormone signal transduction, MAPK signaling pathway-plant, starch and sucrose metabolism, alpha-Linolenic acid metabolism, plant-pathogen interaction, diterpenoid biosynthesis, glucuronate interconversions, phenylpropanoid biosynthesis, compared to 1 h before floret opening. In addition, the expression patterns of some genes, OsJAZ13, OsJAZ11 and OsCML1 which the above metabolic pathways, were different between Indica and Japonica rice. Compared to the floret opening, the differentially expressed genes at floret closure were mainly involved in the following three metabolic pathways: Circadian rhythm-plant, sesquiterpenoid and triterpenoid biosynthesis, starch and sucrose metabolism, and thiamine metabolism. This study provides insights into revealing the molecular mechanism of floret opening and closure in rice. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03226-y.

9.
CBE Life Sci Educ ; 21(3): ar49, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35833940

RESUMO

Infusing undergraduate curricula with authentic research training is an important contemporary challenge. Such exposure typically occurs through mentored research (MR) or course-based undergraduate research experiences (CUREs). In Asian contexts, CURE implementation is rare, while MR is often a graduation requirement. In this study, mentor interviews and mentee focus groups were used to characterize the learning challenges associated with this requirement at a Chinese university. An intensive 6-week CURE was then implemented as an MR preparatory program to help mitigate the identified challenges. This program contained seven site-specific features not typically included in other CUREs, each designed to improve different aspects of student readiness for MR. Post-CURE surveys, focus groups, and interviews demonstrated CURE enrollment significantly improved subsequent MR outcomes. Almost 90% of all enrollees, for example, began their first MR experience in their second year, more than twice the rate of non-enrollees. Enrollees also reported greater confidence in their research skills and more frequent experiences working in multiple labs. This study reports both immediate CURE and downstream MR outcomes, using the former to help explain the latter. A comprehensive CURE implementation process is described, offering a potential model for the design of other programs with similar research enhancement goals.


Assuntos
Biologia/educação , Mentores , Pesquisa/educação , Estudantes , Currículo , Humanos , Aprendizagem , Universidades
10.
Genome Res ; 2022 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-35760561

RESUMO

Clinical exome sequencing has yielded extensive disease-related missense single-nucleotide variants (SNVs) of uncertain significance, leading to diagnostic uncertainty. KCNQ4 is one of the most commonly responsible genes for autosomal dominant nonsyndromic hearing loss. According to the gnomAD cohort, approximately one in 100 people harbors missense variants in KCNQ4 (missense variants with minor allele frequency > 0.1% were excluded), but most are of unknown consequence. To prospectively characterize the function of all 4085 possible missense SNVs of human KCNQ4, we recorded the whole-cell currents using the patch-clamp technique and categorized 1068 missense SNVs as loss of function, as well as 728 loss-of-function SNVs located in the transmembrane domains. Further, to mimic the heterozygous condition in Deafness nonsyndromic autosomal dominant 2 (DFNA2) patients caused by KCNQ4 variants, we coexpressed loss-of-function variants with wild-type KCNQ4 and found 516 variants showed impaired or only partially rescued heterogeneous channel function. Overall, our functional classification is highly concordant with the auditory phenotypes in Kcnq4 mutant mice and the assessments of pathogenicity in clinical variant interpretations. Taken together, our results provide strong functional evidence to support the pathogenicity classification of newly discovered KCNQ4 missense variants in clinical genetic testing.

11.
Can J Gastroenterol Hepatol ; 2022: 6015877, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35719323

RESUMO

Background: Gastric cancer is one of the most common malignant tumors in the world. Albumin paclitaxel (Nab-PTX) is a novel microtubule inhibitor with albumin as the carrier. Several clinical trials are underway in gastric cancer, but the autophagy mechanism of Nab-PTX on gastric cancer is still unclear. The autophagy and apoptosis effects of Nab-PTX compared with 5-pentafluorouracil (5-Fu) and lobaplatin (LBP) in gastric cancer are also unclear. Objective: This article will compare the effects of Nab-PTX, 5-Fu, LBP, and albumin paclitaxel + 5-pentafluorouracil (Nab-PTX + 5-Fu) on AGS cells from the perspective of autophagy and apoptosis, which is to provide new ideas and experimental evidence for gastric cancer. Method: (1) Experimental groups were control (Ctrl), Nab-PTX, 5-Fu, LBP, and Nab-PTX + 5-Fu. (2) CCK-8 assay was used to reflect cell viability and proliferation. (3) The flow cytometry was used to perform the 24-hour apoptosis and cell cycle of each group. (4) Western blot assay was used to investigate autophagy signal proteins LC3I/LC3II, LC3II/LC3I, SQSTM1/p62, Beclin-1, Atg12, Atg5, p-mULK1, p-AMPK, p-mTOR, and apoptosis signal proteins Bax and Bcl-2. Results: Nab-PTX, 5-Fu, LBP, and Nab-PTX + 5-Fu inhibited AGS cells' proliferation and arrested the cell cycle. At the same time, each group increased the apoptosis of AGS cells to various degrees (Nab-PTX + 5-Fu > Nab-PTX > 5-Fu > LBP, respectively). The experimental results showed that Nab-PTX and Nab-PTX + 5-Fu promoted autophagy and apoptosis of AGS cells. The comparison of Nab-PTX, 5-Fu, and LBP between groups revealed that 5-Fu inhibited autophagy and the expression of apoptosis protein Bax. In LBP, abnormal activation of autophagy downstream, blocking of autophagy flow, abnormal increase of ATG12, and increased expression of apoptosis protein Bax occurred. Further study found that the autophagy upstream mechanism is different. Conclusion: Nab-PTX, 5-Fu, LBP, and Nab-PTX + 5-Fu can inhibit cell proliferation, promote cell apoptosis, and induce the difference in autophagy expression. The autophagy difference of this antitumor drug may be related to its inducing apoptosis. Meanwhile, Nab-PTX has a better antitumor effect than 5-Fu and LBP in gastric cancer, and the combination of Nab-PTX + 5-Fu has more antitumor advantages.


Assuntos
Paclitaxel , Neoplasias Gástricas , Albuminas/farmacologia , Albuminas/uso terapêutico , Apoptose , Autofagia , Linhagem Celular Tumoral , Ciclobutanos , Fluoruracila/farmacologia , Humanos , Compostos Organoplatínicos , Paclitaxel/farmacologia , Paclitaxel/uso terapêutico , Neoplasias Gástricas/patologia , Proteína X Associada a bcl-2/farmacologia
12.
Clin Genet ; 102(2): 149-154, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35599357

RESUMO

The pathogenic variants in KCNQ4 cause DFNA2 nonsyndromic hearing loss. However, the understanding of genotype-phenotype correlations between KCNQ4 and hearing is limited. Here, we identified a novel KCNQ4 mutation p.G228D from a Chinese family, including heterozygotes characterized by high-frequency hearing loss that is progressive across all frequencies and homozygotes with more severe hearing loss. We constructed a novel murine model with humanized homologous Kcnq4 mutation. The heterozygotes had mid-frequency and high-frequency hearing loss at 4 weeks, and moved toward all frequencies hearing loss at 12 weeks, while the homozygotes had severe-to-profound hearing loss at 8 weeks. The degeneration of outer hair cells (OHCs) was observed from basal to apical turn of cochlea. The reduced K+ currents and depolarized resting potentials were revealed in OHCs. Remarkably, we observed the loss of inner hair cells (IHCs) in the region corresponding to the frequency above 32 kHz at 8-12 weeks. The results suggest the degeneration of OHCs and IHCs may contribute to high-frequency hearing loss in DFNA2 over time. Our findings broaden the variants of KCNQ4 and provide a novel mouse model of progressive hearing loss, which contributes to an understanding of pathogenic mechanism and eventually treatment of DFNA2 progressive hearing loss.


Assuntos
Perda Auditiva de Alta Frequência , Canais de Potássio KCNQ , Animais , China , Modelos Animais de Doenças , Perda Auditiva de Alta Frequência/genética , Humanos , Canais de Potássio KCNQ/genética , Camundongos , Mutação
13.
J Adult Dev ; : 1-8, 2022 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-35600507

RESUMO

Insecure attachment is regarded as a risk factor for depressive symptoms. However, insecure attachment can be divided into attachment avoidance and anxiety, so a better understanding of the relationship between attachment avoidance, attachment anxiety, and depressive symptoms is needed. The current study applied a structural equation model to verify our hypothesis that different facets of affective empathy mediate the relationship between attachment avoidance, attachment anxiety, and depressive symptoms. The participants of this study included four hundred and sixty-four undergraduate students who completed the Experiences in Close Relationships-Relationship Structures Questionnaire, Interpersonal Reactivity Index, and Center for Epidemiologic Studies Depression Scale. The results indicated that the correlation between attachment avoidance and depressive symptoms was mediated by empathic concern, while the correlation between attachment anxiety and depressive symptoms was mediated by personal distress. These results implied that attachment avoidance and attachment anxiety work on depressive symptoms through different pathways.

14.
Artigo em Inglês | MEDLINE | ID: mdl-35445870

RESUMO

PURPOSE: Chimeric antigen receptor (CAR) T cell therapy has demonstrated clinical success in treating haematologic malignancies but has not been effective against solid tumours thus far. Trop2 is a tumour-related antigen broadly overexpressed on a variety of tumours and has been reported as a promising target for pancreatic cancers. Our study aimed to determine whether CAR T cells designed with a fully human Trop2-specific single-chain fragment variable (scFv) can be used in the treatment of Trop2-positive pancreatic tumours. METHODS: We designed Trop2-targeted chimeric antigen receptor engineered T cells with a novel human anti-Trop2 scFv (2F11) and then investigated the cytotoxicity, degranulation, and cytokine secretion profiles of the anti-Trop2 CAR T cells when they were exposed to Trop2 + cancer cells in vitro. We also studied the antitumour efficacy and toxicity of Trop2-specific CAR T cells in vivo using a BxPC-3 pancreatic xenograft model. RESULTS: Trop2-targeted CAR T cells designed with 2F11 effectively killed Trop2-positive pancreatic cancer cells and produced high levels of cytotoxic cytokines in vitro. In addition, Trop2-targeted CAR T cells, which persistently circulate in vivo and efficiently infiltrate into tumour tissues, significantly blocked and even eliminated BxPC-3 pancreatic xenograft tumour growth without obvious deleterious effects observed after intravenous injection into NSG mice. Moreover, disease-free survival was efficiently prolonged. CONCLUSION: These results show that Trop2-targeted CAR T cells equipped with a fully human anti-Trop2 scFv could be a potential treatment strategy for pancreatic cancer and could be useful for clinical evaluation.

15.
J Hematol Oncol ; 15(1): 39, 2022 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-35365211

RESUMO

Persistence of CAR-T cell function is associated with relapse rate after CAR-T therapy, while co-stimulatory agents are highly concerned with the persistence of CAR-T cells. In this study, we designed and constructed a series of BCMA-targeting second-generation CAR constructs containing CD28, 41BB, and OX40 molecules, respectively, to identify the costimulatory domains most favorable for persistence. The results of routine in vitro studies showed that OX40-CAR-T and 41BB-CAR-T had similar antitumor effects and were superior to CD28-CAR-T in terms of proliferation and cytotoxicity. Although difficult to distinguish by conventional functional assays, OX40-CAR-T cells exhibited greater proliferation and enhanced immune memory than 41BB-CAR-T cells with the repeated stimulation assay by BCMA-expressing target cells. In vivo studies further demonstrated that OX40-CAR-T cells had stronger proliferative activity than 41BB-CAR-T cells, which was highly consistent with the in vitro antitumor activity and proliferation results. Our study provides for the first time a scientific basis for designing OX40-CAR-T cell therapy to improve relapse in patients with MM after CAR-T treatment.


Assuntos
Mieloma Múltiplo , Receptores de Antígenos Quiméricos , Humanos , Imunoterapia Adotiva/métodos , Mieloma Múltiplo/terapia , Recidiva Local de Neoplasia , Linfócitos T
16.
ACS Appl Mater Interfaces ; 14(18): 21453-21460, 2022 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-35486103

RESUMO

Singlet chiral photocatalysis is highly challenging. Herein, we report fluorescence resonance energy transfer (FRET)-based chiral photocatalysis with γ-cyclodextrin (CD)-grafted lanthanide-doped upconverted nanoparticles (UCNP). The CD-modified UCNP strongly emits in the UV wavelength region upon excitation with a 980 nm laser, which selectively sensitizes the photosubstrates complexed by CD on the surface of UCNP through FRET. Therefore, enantiodifferentiating photocyclodimerization of anthracene or naphthalene derivatives sensitized by the CD-modified UCNP gives photoproducts in good enantioselectivity even in the presence of a catalytic amount of CD-modified UCNP. Moreover, the photocatalysts are readily separated and could be reused for at least six cycles without decreasing the enantioselectivity.

17.
RSC Adv ; 12(4): 2375-2382, 2022 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-35425215

RESUMO

A microfluidic-based digital polymerase chain reaction (dPCR) chip requires precise temperature control as well as uniform temperature distribution to ensure PCR efficiency. However, measuring local temperature and its distribution over thousands of µL/nL-volume samples with minimum disturbance is challenging. Here, we present a method of non-contact localized temperature measurement for determination of the non-uniformity of temperature distribution over a dPCR chip. We filled the dPCR chip with a PCR solution containing amplified DNA fragments with a known melting temperature (T M). We then captured fluorescent images of the chip when it was heated from 70 to 99 °C, plotted the fluorescence intensity of each partition as a function of temperature, and calculated measured T M values from each partition. Finally, we created a 3-D map of the dPCR chip with the measured T M as the parameter. Even when the actual T M of the PCR solution was constant, the measured T M value varied between locations due to temperature non-uniformity in the dPCR chip. The method described here thereby characterized the distribution of temperature non-uniformity using a PCR solution with known T M as a temperature sensor. Among the non-contact temperature measurement methods, the proposed T M-based method can determine the temperature distribution within the chip, instead of only at the chip surface. The method also does not suffer from the undesirable photobleaching effect of fluorescein-based temperature measurement method. Temperature determination over the dPCR chip based on T M allowed us to calibrate the temperature sensor and improve the dPCR configuration and precision. This method is also suitable for determining the temperature uniformity of other microarray systems where there is no physical access to the system and thus direct temperature measurement is not possible.

18.
Int J Med Sci ; 19(3): 511-524, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35370461

RESUMO

Background and aims: The miRNA-based post-transcription modification has been extensively studied in hypertension. It however remains elusive how miRNA expression is regulated in this pathological process. We hypothesize that hydroxymethylation in the promoter regions tightly controls the levels of key miRNAs, which in turn affects the development of hypertension. Methods: The levels of hydroxymethylation in the promoter regions from thoracic aortic tissues were compared between spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto rats (WKYs), using hydroxymethylcytosine DNA immunoprecipitation (hMeDIP) sequencing. The altered hydroxymethylation level of miR-3571 was confirmed by glucosylation-coupled hydroxymethylation-sensitive qPCR. We further identified claudin 1(CLDN1) as a key target of miR-3571 via bioinformatic prediction (targetscan) and dual-luciferase activity assays. Finally, we analyzed the contribution of miR-3571/CLDN1 axis in the proliferation and migration of vascular smooth muscle cells (VSMCs). Results: The hydroxymethylation level of miR-3571 promoter region in thoracic aortic tissue from spontaneously hypertensive rats was lower than that from normotensive Wistar-Kyoto rats. Accordingly, the expression of miR-3571 was lower during hypertension, with up-regulated CLDN1 protein levels. More importantly, we found that miR3571 overexpression led to phenotypic changes of VSMCs, and inhibited the proliferation and migration of muscle cells via suppressing CLDN1 as well. Our findings further suggested that CLDN1 up-regulation increase the activity of ERK1/2 in VSMCs. Conclusions: Our study suggested that hydroxymethylation in the promoter regions controlled the level of miR-3571 and revealed the important roles of miR-3571 and CLDN1 in VSMCs during the development of hypertension. In addition, our results also indicated that miR-3571/CLDN1 axis regulated the functions of VSMCs via the ERK1/2 pathway. Taken together, our findings support miR-3571 as a novel biomarker for the diagnosis and prevention of hypertension.


Assuntos
MicroRNAs , Músculo Liso Vascular , Animais , Movimento Celular/genética , Proliferação de Células/genética , Claudina-1/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Músculo Liso Vascular/metabolismo , Ratos , Ratos Endogâmicos WKY
19.
Bioengineered ; 13(3): 6243-6256, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35349390

RESUMO

Plentiful studies have clarified that circular RNAs (circRNAs) are crucial in colorectal cancer (CRC)'s occurrence and development, but its function has not been fully elucidated. The purpose of this study was to investigate the biological functions of circPLCE1 on epithelial mesenchymal transformation (EMT) and glycolysis in CRC, and tumor-associated macrophage (TAM) polarization. The results affirmed augment of circPLCE1 and γ-Actin Gene (ACTG1) but decline of miR-485-5p in CRC. Knockdown circPCLE1 refrained CRC proliferation, glucose consumption, lactic acid and pyruvate production, M2 macrophage markers (IL-10, MRC1), N-cadherin, Snail, reduced the proportion of CD206+ and CD168+ macrophages, but expedited M1 macrophage markers (TNF-α, IL-6) and E-cadherin, while descending miR-485-5p expedited EMT, glycolysis in CRC and TAM M2 polarization . Additionally, it was affirmed that the repression or motivation of depressive or elevated circPCLE1 on EMT, glycolysis in CRC and TAM M2 polarization were reversed via facilitated ACTG1 and miR-485-5p, separately. Mechanism studies have clarified that circPCLE1 as a competitive endogenous RNA adsorbed miR-485-5p to mediate ACTG1. It was assured that refrained circPCLE1 constrained CRC tumor growth, EMT and TAM M2 polarization. In brief, circPCLE1 expedites EMT, glycolysis in CRC and TAM M2 polarization via modulating the miR-485-5p/ACTG1 axis, and is supposed to be a latent molecular target for CRC therapy later.


Assuntos
Neoplasias Colorretais , MicroRNAs , Proliferação de Células/genética , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Transição Epitelial-Mesenquimal/genética , Glicólise/genética , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Circular/genética , Macrófagos Associados a Tumor
20.
Mol Ther Oncolytics ; 24: 719-728, 2022 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-35317521

RESUMO

Although chimeric antigen receptor (CAR) T cell immunotherapy has shown promising significance in B cell malignancies, success against T cell malignancies remains unsatisfactory because of shared antigenicity between normal and malignant T cells, resulting in fratricide and hindering CAR production for clinical treatment. Here, we report a new strategy of blocking the CD7 antigen on the T cell surface with a recombinant anti-CD7 antibody to obtain a sufficient amount of CD7-targeting CAR-T cells for T cell acute lymphoblastic leukemia (T-ALL) treatment. Feasibility was evaluated systematically, revealing that blocking the CD7 antigen with an antibody effectively blocked CD7-derived fratricide, increased the expansion rate, reduced the proportion of regulatory T (Treg) cells, maintained the stem cell-like characteristics of T cells, and restored the proportion of the CD8+ T cell population. Ultimately, we obtained anti-CD7 CAR-T cells that were specifically and effectively able to kill CD7 antigen-positive target cells, obviating the need for complex T cell modifications. This approach is safer than previous methods and provides a new, simple, and feasible strategy for clinical immunotherapies targeting CD7-positive malignant tumors.

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