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1.
Nan Fang Yi Ke Da Xue Xue Bao ; 41(10): 1456-1463, 2021 Oct 20.
Artigo em Chinês | MEDLINE | ID: mdl-34755660

RESUMO

OBJECTIVE: To explore the association of methylation levels of C19orf57, MAP9, EMR3, NEK6 and PCOLCE2 genes in peripheral blood with breast cancer (BC) in Chinese women. METHODS: We collected peripheral blood samples from 258 early-stage BC patients and 272 healthy women. Agena matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was utilized to quantitatively measure the methylation levels of CpG sites in the genes. The association between DNA methylation and BC was analyzed using a logistic regression model adjusted for covariants. Spearman's correlation analysis was performed to analyze the association between the gene methylation levels and age. The methylation levels of the genes in the BC patients with different clinical characteristics were investigated using non-parametric tests. RESULTS: In stead of EMR3 gene hypermethylation as found in BC patients as found in the Caucasian population, EMR3 gene hypomethylation was found to correlate with BC in Chinese women, but this correlation was significant only in women beyond the age of 50 years (for every 10% reduction of the methylation level, EMR3_CpG_1: OR=1.40; EMR3_CpG_2: OR=2.31; EMR3_CpG_3: OR=2.76, P < 0.05). EMR3 methylation was not or was only weakly correlated with tumor stage, size, lymphatic metastasis, ER, PR, HER2, or Ki67. Our data did not show a correlation between C19orf57 methylation and BC. CONCLUSION: Peripheral blood EMR3 gene hypomethylation is associated with BC in Chinese women, especially in those at an old age and in postmenopausal women.


Assuntos
Neoplasias da Mama , Neoplasias da Mama/genética , China , Ilhas de CpG/genética , Metilação de DNA , Feminino , Humanos , Metástase Linfática , Proteínas Associadas aos Microtúbulos , Pessoa de Meia-Idade , Quinases Relacionadas a NIMA
2.
J Transl Med ; 19(1): 459, 2021 11 07.
Artigo em Inglês | MEDLINE | ID: mdl-34743703

RESUMO

BACKGROUND: Adoptive transfer of chimeric antigen receptor (CAR)-engineered T cells combined with checkpoint inhibition may prevent T cell exhaustion and improve clinical outcomes. However, the approach is limited by cumulative costs and toxicities. METHODS: To overcome this drawback, we created a CAR-T (RB-340-1) that unites in one product the two modalities: a CRISPR interference-(CRISPRi) circuit prevents programmed cell death protein 1 (PD-1) expression upon antigen-encounter. RB-340-1 is engineered to express an anti-human epidermal growth factor receptor 2 (HER2) CAR single chain variable fragment (scFv), with CD28 and CD3ζ co-stimulatory domains linked to the tobacco etch virus (TEV) protease and a single guide RNA (sgRNA) targeting the PD-1 transcription start site (TSS). A second constructs includes linker for activation of T cells (LAT) fused to nuclease-deactivated spCas9 (dCas9)-Kruppel-associated box (KRAB) via a TEV-cleavable sequence (TCS). Upon antigen encounter, the LAT-dCas9-KRAB (LdCK) complex is cleaved by TEV allowing targeting of dCas9-KRAB to the PD-1 gene TSS. RESULTS: Here, we show that RB-340-1 consistently demonstrated higher production of homeostatic cytokines, enhanced expansion of CAR-T cells in vitro, prolonged in vivo persistence and more efficient suppression of HER2+ FaDu oropharyngeal cancer growth compared to the respective conventional CAR-T cell product. CONCLUSIONS: As the first application of CRISPRi toward a clinically relevant product, RB-340-1 with the conditional, non-gene editing and reversible suppression promotes CAR-T cells resilience to checkpoint inhibition, and their persistence and effectiveness against HER2-expressing cancer xenografts.

3.
Nat Commun ; 12(1): 6784, 2021 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-34811372

RESUMO

The control of the in-plane domain evolution in ferroelectric thin films is not only critical to understanding ferroelectric phenomena but also to enabling functional device fabrication. However, in-plane polarized ferroelectric thin films typically exhibit complicated multi-domain states, not desirable for optoelectronic device performance. Here we report a strategy combining interfacial symmetry engineering and anisotropic strain to design single-domain, in-plane polarized ferroelectric BaTiO3 thin films. Theoretical calculations predict the key role of the BaTiO3/PrScO3 [Formula: see text] substrate interfacial environment, where anisotropic strain, monoclinic distortions, and interfacial electrostatic potential stabilize a single-variant spontaneous polarization. A combination of scanning transmission electron microscopy, piezoresponse force microscopy, ferroelectric hysteresis loop measurements, and second harmonic generation measurements directly reveals the stabilization of the in-plane quasi-single-domain polarization state. This work offers design principles for engineering in-plane domains of ferroelectric oxide thin films, which is a prerequisite for high performance optoelectronic devices.

4.
Zhonghua Liu Xing Bing Xue Za Zhi ; 42(5): 928-934, 2021 May 10.
Artigo em Chinês | MEDLINE | ID: mdl-34814491

RESUMO

Objective: To better promote the standardization of public health management in China, and provide evidence for the development and improvement of the standardization strategy and management system in public health field in China. Methods: This paper summarizes and analyzes the information about the standardized management mechanism collected from international organizations related with standardization in public health. Results: The standards in public health varied in different management systems of the international organizations, and there were great differences in organization nature, standard types, application, release, organization structure, standard development principles, advantages, transformation, promotion and implementation, and evaluation. Conclusion: China can benefit from the studying of the working mechanism of the international organization related with standardization in public health to facilitate its own standardization in public health.


Assuntos
Administração em Saúde Pública , Saúde Pública , China , Humanos , Padrões de Referência
5.
Elife ; 102021 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-34499031

RESUMO

The type V-A Cas12a protein can process its CRISPR array, a feature useful for multiplexed gene editing and regulation. However, CRISPR arrays often exhibit unpredictable performance due to interference between multiple guide RNA (gRNAs). Here, we report that Cas12a array performance is hypersensitive to the GC content of gRNA spacers, as high-GC spacers can impair activity of the downstream gRNA. We analyze naturally occurring CRISPR arrays and observe that natural repeats always contain an AT-rich fragment that separates gRNAs, which we term a CRISPR separator. Inspired by this observation, we design short, AT-rich synthetic separators (synSeparators) that successfully remove the disruptive effects between gRNAs. We further demonstrate enhanced simultaneous activation of seven endogenous genes in human cells using an array containing the synSeparator. These results elucidate a previously underexplored feature of natural CRISPR arrays and demonstrate how nature-inspired engineering solutions can improve multi-gene control in mammalian cells.


Assuntos
Proteínas de Bactérias/genética , Proteínas Associadas a CRISPR/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Endodesoxirribonucleases/genética , Regulação da Expressão Gênica , Proteínas de Fluorescência Verde/genética , Células HEK293 , Humanos , Óperon , RNA Guia/genética
6.
Mol Cell ; 81(20): 4333-4345.e4, 2021 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-34480847

RESUMO

Compact and versatile CRISPR-Cas systems will enable genome engineering applications through high-efficiency delivery in a wide variety of contexts. Here, we create an efficient miniature Cas system (CasMINI) engineered from the type V-F Cas12f (Cas14) system by guide RNA and protein engineering, which is less than half the size of currently used CRISPR systems (Cas9 or Cas12a). We demonstrate that CasMINI can drive high levels of gene activation (up to thousands-fold increases), while the natural Cas12f system fails to function in mammalian cells. We show that the CasMINI system has comparable activities to Cas12a for gene activation, is highly specific, and allows robust base editing and gene editing. We expect that CasMINI can be broadly useful for cell engineering and gene therapy applications ex vivo and in vivo.

7.
Artigo em Inglês | MEDLINE | ID: mdl-34425228

RESUMO

OBJECTIVE: There is considerable evidence for relationship between gut microbiota and osteoarthritis (OA), but no studies have investigated their causal relationship. METHOD: This study utilized large-scale genome-wide association studies (GWAS) summary statistics to evaluate the causal association between gut microbiota and OA risk. Specifically, two-sample Mendelian randomization (MR) approach was used to identify the causal microbial taxa for OA. Comprehensively sensitive analyses were performed to validate the robustness of results and novel multivariable MR analyses were further conducted to ensure the independence of causal association. Reverse-direction MR analyses were performed to rule out the possibility of reverse associations. Finally, enrichment analyses were used to investigate the biofunction. RESULTS: After correction, three microbial taxa were identified to be causally associated with diverse joint OA (PFDR < 0.100), namely Methanobacteriaceae family for knee OA (PFDR = 0.043) and any OA (PFDR = 0.028), Desulfovibrionales order for knee OA (PFDR = 0.045) and Ruminiclostridium5 genus for knee OA (PFDR = 0.063). In addition, we also identified five suggestive microbial taxa that were significant with three different methods under the nominal significance (P < 0.05). Sensitive analysis excluded the influence of heterogeneity and horizontal pleiotropy and multivariable MR analysis ruled out the possibility of horizontal pleiotropy of BMI. GO enrichment analysis illustrates the protective mechanism of the identified taxa against OA. CONCLUSIONS: This study found that several microbial taxa were causally associated with diverse joint OA. The results enhanced our understanding of gut microbiota in the pathology of OA.

8.
Cell Rep Med ; 2(7): 100343, 2021 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-34337559

RESUMO

Regenerative medicine approaches utilizing stem cells offer a promising strategy to address tendinopathy, a class of common tendon disorders associated with pain and impaired function. Tendon progenitor cells (TPCs) are important in healing and maintaining tendon tissues. Here we provide a comprehensive single cell transcriptomic profiling of TPCs from three normal and three clinically classified tendinopathy samples in response to mechanical stimuli. Analysis reveals seven distinct TPC subpopulations including subsets that are responsive to the mechanical stress, highly clonogenic, and specialized in cytokine or growth factor expression. The single cell transcriptomic profiling of TPCs and their subsets serves as a foundation for further investigation into the pathology and molecular hallmarks of tendinopathy in mechanical stimulation conditions.

9.
Mol Cell ; 81(20): 4287-4299.e5, 2021 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-34428454

RESUMO

Eukaryotic chromosomes feature large regions of compact, repressed heterochromatin hallmarked by Heterochromatin Protein 1 (HP1). HP1 proteins play multi-faceted roles in shaping heterochromatin, and in cells, HP1 tethering to individual gene promoters leads to epigenetic modifications and silencing. However, emergent properties of HP1 at supranucleosomal scales remain difficult to study in cells because of a lack of appropriate tools. Here, we develop CRISPR-engineered chromatin organization (EChO), combining live-cell CRISPR imaging with inducible large-scale recruitment of chromatin proteins to native genomic targets. We demonstrate that human HP1α tiled across kilobase-scale genomic DNA form novel contacts with natural heterochromatin, integrates two distantly targeted regions, and reversibly changes chromatin from a diffuse to compact state. The compact state exhibits delayed disassembly kinetics and represses transcription across over 600 kb. These findings support a polymer model of HP1α-mediated chromatin regulation and highlight the utility of CRISPR-EChO in studying supranucleosomal chromatin organization in living cells.

10.
Artigo em Chinês | MEDLINE | ID: mdl-34365769

RESUMO

Objective: To investigate the distribution of noise exposure between non-steady state noise and steady-state noise for metal processing workers, which will provide scientific basis for the prevention and treatment of noise hazards in metal processing industry. Methods: The cross-section method was used to investigate the noise exposure of 737 workers from three metal processing industries in Zhejiang Province from October to December 2017. The general demographic information and occupational history were collected by questionnaire. The noise was recorded by individual noise meters, and the noise exposure intensity (equivalent continuous A-weighted noise exposure level normalized to an 8 h-working-day, L(Aeq, 8 h)) and kurtosis were calculated. Results: Workers exposed to noise in the metal processing industry were mainly 18-40 years old (527 workers, 71.51%) , men (570 workers, 77.34%) , and junior high school education background (416 workers, 56.45%) . There were 572 workers (77.61%) with noise exposure intensity (L(Aeq, 8 h)) greater than 85 dB (A) , 558 workers (75.71%) exposed to non-steady state noise (kurtosis ≥4) , and 634 workers (86.02%) with exposure duration less than 8 years. Among the 30 work types investigated, the work types with noise intensity reaching 100% were the stamping, welding and others from a children's car manufacturing factory in Ningbo, operating, chamfering, tapping, and thread rolling from an automobile parts manufacturing factory in Ningbo; The work types with a rate of 100% exposed to non-steady state noise were the grinding from a children's car manufacturing factory in Ningbo, assembling, assembly operating and others from an automobile brake manufacturing factory in Wenzhou, and polishing from an automobile parts manufacturing factory in Ningbo. Conclusion: Metal processing workers have a high rate of over-standard exposed to high noise intensity and a high proportion exposed to non-steady state noise. It is necessary to take sound insulation and noise reduction engineering control, and strengthen personal protection and occupational health management measures to prevent and control the noise hazards.


Assuntos
Perda Auditiva Provocada por Ruído , Ruído Ocupacional , Exposição Ocupacional , Adolescente , Adulto , Criança , Perda Auditiva Provocada por Ruído/epidemiologia , Perda Auditiva Provocada por Ruído/etiologia , Humanos , Masculino , Indústria Manufatureira , Inquéritos e Questionários , Adulto Jovem
11.
Zhonghua Fu Chan Ke Za Zhi ; 56(7): 489-497, 2021 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-34304441

RESUMO

Objective: To investigate the correlations of laminin subunit gamma 3 (LAMC3) expression with prognosis of ovarian cancer (OC). Methods: LAMC3 protein expression was measured using immunohistochemical streptavidin-peroxidase-biotin connection method (IHC). Gene expression and related clinical data in the cancer genome atlas (TCGA) cohort and clinical proteomic tumor analysis consortium (CPTAC) were applied to analyse the correlation between gene and protein expressions and clinical outcomes. Correlations between LAMC3 and clinicopathological factors were evaluated using the Pearson χ2 test (2-sided). The probability of survival and significance was calculated using the Kaplan-Meier plot. The functional clustering of biological pathways enriched from co-expressed genes of LAMC3 was used to explore the possible mechanisms that LAMC3 might contribute to poor prognosis. Results: Based on the IHC results of 216 OC tissues or ovaries (including 208 tumors and 8 normal tissues) and 51 OC tissues (including 24 chemotherapy-resistant and 27 sensitive tissues), and the protein expression data from CPTAC (including 100 primary tumors and 25 normal tissues), the results showed that the protein expression of LAMC3 was significantly decreased in OC tissues compared with normal, decreased in advanced-stage tissues compared with early-stage tissues, and decreased in drug-resistant tissues compared with sensitive tissues (all P<0.05). Furthermore, low expression of LAMC3 protein was significantly associated with poor disease-free survival (DFS) and overall survival (OS) in 51 OC tissues (P<0.01), consistent with the results that the low levels of LAMC3 mRNA predicted short DFS and OS in 489 OC tissues of the TCGA cohort (P<0.05). The results suggested that low expression of LAMC3 might be the adverse factors for OC development, such as drug resistance and advanced tumors, and might be a risk indicator for prognosis. Moreover, functional clustering of biological pathways enriched from the co-expressed genes of LAMC3 in TCGA ovarian cohort indicated that LAMC3 potentially involved in regulation of OC via oncogene-pathways such as Ras associated protein 1 (Rap1), mitogen-activated protein kinase (MAPK), Ras and cell adhesion-related pathways such as extra cellular matrix (ECM)-receptor interaction and focal adhesion. It indicated that LAMC3 might contribute to short survival and tumor progression by regulation of the above pathways. Conclusion: Low expression of LAMC3 is related to poor prognosis and malignant progression in OC, and thus it is expected to be a new prognostic marker and therapeutic target for clinical treatment.


Assuntos
Neoplasias Ovarianas , Proteômica , Biomarcadores Tumorais/genética , Carcinoma Epitelial do Ovário , Feminino , Humanos , Laminina , Neoplasias Ovarianas/genética , Prognóstico
12.
Nan Fang Yi Ke Da Xue Xue Bao ; 41(7): 1079-1086, 2021 Jul 20.
Artigo em Chinês | MEDLINE | ID: mdl-34308860

RESUMO

OBJECTIVE: To study the response characteristics of the secondary auditory cortex (A2) to wriggling calls (WC) and the mechanism of response modulation in female mice. METHODS: We used patch-clamp and immunofluorescence labeling technique to mark and record the action potential and cell type of A2 neurons. Female C57 mice were stimulated with pure tone and white noise (control), 4.5 kHz, 9 kHz, or 13.5 kHz sound waves extracted from WC (single-frequency simulation group), the combinations of every two of the 3 single-frequency sound waves (two-tone frequency simulation group), or the combinations of 4.5 kHz+7.7 kHz+13.5 kHz and 4.5 kHz+9 kHz+13.5 kHz sound waves (three-frequency simulation group). The firing pattern, firing number, threshold, and latency of the action potential of the A2 neurons were recorded in response to the stimulations. RESULTS: By comparison of the spikes elicited by different sound stimulations, we identified 3 types of neurons with different sensitivities to WC. The WC-sensitive neurons had a significantly greater number of spikes in response to WC than to other sounds and noise (P < 0.001). Comparison of the latency and threshold revealed significantly longer latencies of the WC-sensitive neurons and WC-insensitive neurons in response to WC stimulation than to pure tone stimulation (P=0.002), but their latencies to WC and noise stimulation were similar (P=0.093). The WC-sensitive neurons also had lower threshold to WC than to pure tone and noise stimulations (P=0.02). Analysis of the firing patterns of action potentials showed that the WCsensitive neurons consisted mainly of parvalbumin interneurons. The results of immunofluorescence labeling indicated that inhibitory interneurons were present in the A2 neurons that responded to WC. CONCLUSION: A2 contains 3 types of neurons with different sensitivities to WC. Among them, the WC-sensitive neurons is mainly PV neurons, whose response characteristics to different types of sounds can help to explain the mechanism of communication sound recognition and response modification in A2.


Assuntos
Córtex Auditivo , Estimulação Acústica , Potenciais de Ação , Animais , Feminino , Camundongos , Neurônios , Som
13.
Methods Mol Biol ; 2320: 261-281, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34302664

RESUMO

Identifying causative genes in a given phenotype or disease model is important for biological discovery and drug development. The recent development of the CRISPR/Cas9 system has enabled unbiased and large-scale genetic perturbation screens to identify causative genes by knocking out many genes in parallel and selecting cells with desired phenotype of interest. However, compared to cancer cell lines, human somatic cells including cardiomyocytes (CMs), neuron cells, and endothelial cells are not easy targets of CRISPR screens because CRISPR screens require a large number of isogenic cells to be cultured and thus primary cells from patients are not ideal. The combination of CRISPR screens with induced pluripotent stem cell (iPSC) technology would be a powerful tool to identify causative genes and pathways because iPSCs can be expanded easily and differentiated to any cell type in principle. Here we describe a robust protocol for CRISPR screening using human iPSCs. Because each screening is different and needs to be customized depending on the cell types and phenotypes of interest, we show an example of CRISPR knockdown screening using CRISPRi system to identify essential genes to differentiate iPSCs to CMs.


Assuntos
Sistemas CRISPR-Cas , Edição de Genes/métodos , Células-Tronco Pluripotentes Induzidas/citologia , Sequência de Bases , Causalidade , Células Cultivadas , Cromatografia Líquida/métodos , DNA/isolamento & purificação , Doxiciclina/farmacologia , Citometria de Fluxo , Estudos de Associação Genética , Vetores Genéticos/genética , Células HEK293 , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Lentivirus/genética , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , RNA Guia/genética , Transfecção
14.
Zhonghua Zhong Liu Za Zhi ; 43(7): 756-761, 2021 Jul 23.
Artigo em Chinês | MEDLINE | ID: mdl-34289569

RESUMO

Cancer, one of the major public health problems in the world, threatens human health seriously, and the burden of disease is heavy. Disability adjusted life years (DALYs) have been increasingly used to estimate the burden of disease worldwide. Disability weights is a key ingredient for estimating DALYs, and its value directly affects the calculation of disease burden. In this review, we summarize the research methods, key issues, and progress on disability weights for cancer both domestic and abroad, in order to provide valuable information for the estimation of cancer disability weights in China.


Assuntos
Pessoas com Deficiência , Neoplasias , China , Efeitos Psicossociais da Doença , Humanos , Anos de Vida Ajustados por Qualidade de Vida
15.
bioRxiv ; 2021 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-34127974

RESUMO

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the third human coronavirus within 20 years that gave rise to a life-threatening disease and the first to reach pandemic spread. To make therapeutic headway against current and future coronaviruses, the biology of coronavirus RNA during infection must be precisely understood. Here, we present a robust and generalizable framework combining high-throughput confocal and super-resolution microscopy imaging to study coronavirus infection at the nanoscale. Employing the model human coronavirus HCoV-229E, we specifically labeled coronavirus genomic RNA (gRNA) and double-stranded RNA (dsRNA) via multicolor RNA-immunoFISH and visualized their localization patterns within the cell. The exquisite resolution of our approach uncovers a striking spatial organization of gRNA and dsRNA into three distinct structures and enables quantitative characterization of the status of the infection after antiviral drug treatment. Our approach provides a comprehensive framework that supports investigations of coronavirus fundamental biology and therapeutic effects.

16.
Nat Commun ; 12(1): 2397, 2021 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-33893274

RESUMO

Gene targeting studies in primary human islets could advance our understanding of mechanisms driving diabetes pathogenesis. Here, we demonstrate successful genome editing in primary human islets using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9). CRISPR-based targeting efficiently mutated protein-coding exons, resulting in acute loss of islet ß-cell regulators, like the transcription factor PDX1 and the KATP channel subunit KIR6.2, accompanied by impaired ß-cell regulation and function. CRISPR targeting of non-coding DNA harboring type 2 diabetes (T2D) risk variants revealed changes in ABCC8, SIX2 and SIX3 expression, and impaired ß-cell function, thereby linking regulatory elements in these target genes to T2D genetic susceptibility. Advances here establish a paradigm for genetic studies in human islet cells, and reveal regulatory and genetic mechanisms linking non-coding variants to human diabetes risk.


Assuntos
Sistemas CRISPR-Cas , Edição de Genes/métodos , Células Secretoras de Insulina/metabolismo , Ilhotas Pancreáticas/metabolismo , Modelos Genéticos , Sequência de Bases , Diabetes Mellitus Tipo 2/genética , Regulação da Expressão Gênica , Proteínas de Homeodomínio/genética , Humanos , Células Secretoras de Insulina/citologia , Ilhotas Pancreáticas/citologia , Canais de Potássio Corretores do Fluxo de Internalização/genética , Transativadores/genética
17.
Int Endod J ; 54(8): 1328-1341, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33715185

RESUMO

AIM: To profile molecular changes in lipopolysaccharide (LPS)-induced experimental pulpitis in a rat model and explore the feasibility of a molecular-based diagnostic strategy for pulpitis. METHODOLOGY: Seventy-three maxillary incisors of Sprague-Dawley rats were used to establish pulpitis models with LPS. Inflammatory grading was performed in four equal sections of the pulp divided from the injured site to the root apex. An antibody array was used to compare the expression of 67 molecules between control pulp and inflamed pulp 12 and 72 h after LPS application. The levels of differentially expressed molecules in the control and inflamed pulp (collected at 3, 6, 9, 12, 24 and 72 h after LPS treatment) were examined via ELISA, and correlations between inflammatory scores and molecule expression were assessed. The molecule distributions in the pulp were investigated by immunofluorescence staining. Data were analysed with paired t-test, one-way anova, Kruskal-Wallis tests, and Spearman's and Pearson's correlations with significance set at P < 0.05. RESULTS: Polymorphonuclear neutrophils were observed in the injured site 3 h after LPS stimulation. Inflammatory infiltration peaked at 12 h and was limited to the injured site with osteodentine deposition at 72 h. Thirteen molecules were significantly differentially expressed between the control and LPS-injured pulp. ELISA validated that tissue inhibitor of metalloproteinase-1 (TIMP-1) expression dramatically peaked at 12 h (compared with other time points, P < 0.05) and returned to baseline at 72 h. The TIMP-1 concentration was strongly correlated with inflammation severity in the apical three-quarters of the pulp, and the strongest correlation was found in the lower-middle quarter (r = 0.786, P < 0.001). Immunofluorescence staining revealed that in the apical three-quarters of the pulp, TIMP-1 expression was significantly higher in the 12 h group than in the control and 3, 6, 24 and 72 h groups (P < 0.01). CONCLUSION: This study provides a molecular profile of LPS-induced pulpitis in a rat model. TIMP-1 had a strong positive correlation with the severity of dental pulp inflammation, verifying the feasibility of applying biomarkers to identify specific pathological conditions in pulpitis.


Assuntos
Pulpite , Animais , Biomarcadores , Polpa Dentária , Mediadores da Inflamação , Ratos , Ratos Sprague-Dawley , Inibidor Tecidual de Metaloproteinase-1
18.
Zhonghua Nei Ke Za Zhi ; 60(4): 331-337, 2021 Apr 01.
Artigo em Chinês | MEDLINE | ID: mdl-33765702

RESUMO

Objective: To investigate the current situation of insomnia in patients with acute coronary syndrome (ACS), and analyze the influencing factors of insomnia in the ACS patients, so as to provide information on the development of new strategies for the treatment of insomnia in ACS patients. Methods: This is a multicenter and prospective observational study. A total of 771 ACS patients who met the criteria were selected from March 2013 to June 2015. The baseline social demographic information, sleep quality questionnaire, general anxiety disorder scale-7(GAD-7),patient health questionnaire-9(PHQ-9), short-form 12 health survey questionnaire(SF-12), and enhancing recovery in coronary heart disease patients social inventory(ESSI) were completed within 7 days after admission. Logistic regression analyses were used to analyze the influencing factors of insomnia in ACS patients. Results: A total of 741 subjects with valid questionnaires were collected, including 510 males (68.8%) and 231 females (31.2%). Among them, 487 (65.7%) subjects had at least one insomnia symptom: 308 (41.6%) subjects had difficulty in falling asleep, 369 (49.8%) subjects were easy to wake at night, 116 (15.7%) subjects woke up earlier than they expected, 74 (10.0%) subjects experienced both woke up earlier and difficulty in falling asleep, and 53 (7.2%) subjects woke up earlier, woke up at night and had difficulty in falling asleep at the same time. Logistic regression analyses showed that before admission physical activity (OR =0.636, 95%CI 0.411-0.984), depression (OR=1.908, 95%CI 1.101-3.305) and low social support (OR=0.278, 95%CI 1.198-3.301) were independent factors of insomnia in ACS patients. Conclusions: Nearly 2/3 ACS patients have symptoms of insomnia. Difficulty in falling asleep and easy to wake up at night are the most common manifestations. Physical activity, depression and social support independently are associated with insomnia.


Assuntos
Síndrome Coronariana Aguda , Doença das Coronárias , Distúrbios do Início e da Manutenção do Sono , Síndrome Coronariana Aguda/complicações , Síndrome Coronariana Aguda/epidemiologia , Feminino , Inquéritos Epidemiológicos , Humanos , Masculino , Distúrbios do Início e da Manutenção do Sono/complicações , Distúrbios do Início e da Manutenção do Sono/epidemiologia , Inquéritos e Questionários
19.
Cell Rep Med ; 2(4): 100245, 2021 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-33778788

RESUMO

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) and variants has led to significant mortality. We recently reported that an RNA-targeting CRISPR-Cas13 system, called prophylactic antiviral CRISPR in human cells (PAC-MAN), offered an antiviral strategy against SARS-CoV-2 and influenza A virus. Here, we expand in silico analysis to use PAC-MAN to target a broad spectrum of human- or livestock-infectious RNA viruses with high specificity, coverage, and predicted efficiency. Our analysis reveals that a minimal set of 14 CRISPR RNAs (crRNAs) is able to target >90% of human-infectious viruses across 10 RNA virus families. We predict that a set of 5 experimentally validated crRNAs can target new SARS-CoV-2 variant sequences with zero mismatches. We also build an online resource (crispr-pacman.stanford.edu) to support community use of CRISPR-Cas13 for broad-spectrum RNA virus targeting. Our work provides a new bioinformatic resource for using CRISPR-Cas13 to target diverse RNA viruses to facilitate the development of CRISPR-based antivirals.

20.
Nat Rev Genet ; 22(6): 343-360, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33558716

RESUMO

Cancers and developmental disorders are associated with alterations in the 3D genome architecture in space and time (the fourth dimension). Mammalian 3D genome organization is complex and dynamic and plays an essential role in regulating gene expression and cellular function. To study the causal relationship between genome function and its spatio-temporal organization in the nucleus, new technologies for engineering and manipulating the 3D organization of the genome have been developed. In particular, CRISPR-Cas technologies allow programmable manipulation at specific genomic loci, enabling unparalleled opportunities in this emerging field of 3D genome engineering. We review advances in mammalian 3D genome engineering with a focus on recent manipulative technologies using CRISPR-Cas and related technologies.


Assuntos
Sistemas CRISPR-Cas , Núcleo Celular/genética , Edição de Genes , Engenharia Genética/métodos , Genoma , Genômica/métodos , Animais , Humanos
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