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2.
Talanta ; 272: 125756, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38364562

RESUMO

Various nucleic acid molecular machines have emerged in recent years. However, when the nucleic acid tracks are fully depleted, these walkers are highly susceptible to premature release or stalling in regions where the tracks are locally exhausted. In this work, a molecular walking machine with a cleat domain preventing dissociation from the track was explored for ultrasensitive detection of miRNA. It has been verified that the cleat design can enhance the signal amplification efficiency of molecular walking machines for electrochemical miRNA-141 detection. Notably, the single-step electrochemical biosensing platform utilizing the cleat-equipped molecular walking machine (CMWM) is exceptionally straightforward and rapid, concluding the reaction within 90 min and achieving a remarkable low detection limit of 0.26 fM. The proposed molecular walking machine with this specific cleat structure was utilized for the identification of miRNA-141 in cellular lysates, exhibiting remarkable selectivity and consistent reproducibility, showcasing its effective utility in bioanalysis. Therefore, the cleat walker developed in this study introduces an innovative method for constructing a miRNA electrochemical biosensing platform, offering new perspectives for its application in biomolecule detection and clinical disease diagnosis.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Reprodutibilidade dos Testes , Técnicas Biossensoriais/métodos , MicroRNAs/análise , Técnicas Eletroquímicas/métodos , Limite de Detecção
3.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 41(1): 52-58, 2024 Jan 10.
Artigo em Chinês | MEDLINE | ID: mdl-38171560

RESUMO

OBJECTIVE: To develop a genotyping method for the Junior blood type and report on a rare blood type with Jr(a-). METHODS: Healthy O-type RhD+ volunteer donors of the Shenzhen Blood Center from January to May 2021 (n = 1 568) and a pedigree with difficult cross-matching (n = 3) were selected as the study subjects. Serological methods were used for proband's blood type identification, unexpected antibody identification, and antibody titer determination. Polymerase chain reaction-sequence specific primer (PCR-SSP) method was used for typing the proband's RhD gene. ABCG2 gene coding region sequencing and a PCR-SSP genotyping method were established for determining the genotypes of the proband and his family members and screening of Jra antigen-negative rare blood type among the 1 568 blood donors. RESULTS: The proband's ABO and RhD blood types were respectively determined as B and partial D (RHDDVI.3/RHD01N.01), Junior blood type Jra antigen was negative, and plasma had contained anti-D and anti-Jra. Sequencing of the ABCG2 gene revealed that the proband's genotype was ABGG201N.01/ABGG201N.01 [homozygous c.376C>T (p.Gln126X) variants], which is the most common Jr(a-) blood type allele in the Asian population. Screening of the voluntary blood donors has detected no Jr(a-) rare blood type. Statistical analysis of the heterozygotes suggested that the allelic frequency for ABCG2*01N.01 (c.376T) was 0.45%, and the frequency of Jr(a-) rare blood type with this molecular background was about 0.2‰. CONCLUSION: A very rare case of partial DVI.3 type and Jr(a-) rare blood type has been identified. And a method for identifying the Junior blood type through sequencing the coding regions of the ABCG2 gene and PCR-SSP has been established.


Assuntos
Antígenos de Grupos Sanguíneos , Humanos , Antígenos de Grupos Sanguíneos/genética , Genótipo , Técnicas de Genotipagem , Heterozigoto , Alelos , Doadores de Sangue , Sistema do Grupo Sanguíneo Rh-Hr/genética
4.
Toxicol In Vitro ; 96: 105784, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38242296

RESUMO

OBJECTIVE: Endoplasmic reticulum (ER) stress mediates Cd-caused germ cell apoptosis in testis. The effects of 4-phenylbutyric acid (PBA), a classical chaperone, were investigated on Cd-induced apoptosis in mouse GC-1 spermatogonia cells. METHODS: The cells were pretreated with PBA before Cd exposure. TUNEL and flow cytometry assays were applied to determine apoptosis. Some key biomarkers of ER stress were analyzed using RT-PCR and western blot. RESULTS: as expected, the apoptotic cells exposed to Cd apparently increased. The mRNA and protein expression levels of GRP78 and ATF6α, were elevated in the Cd groups. Additional experiments displayed that Cd notably increased IRE1α and JNK phosphorylation, and upregulated XBP-1 mRNA and protein expression. Moreover, p-eIF2α and CHOP expressions were clearly elevated in the Cd groups. Interestingly, PBA almost completely inhibited ER stress and protected spermatogonia against apoptosis induced by Cd. CONCLUSION: PBA alleviated Cd-induced ER stress and spermatogonia apoptosis, and may have the therapeutic role in Cd-induced male reproductive toxicity.


Assuntos
Cádmio , Fenilbutiratos , Espermatogônias , Camundongos , Masculino , Animais , Cádmio/toxicidade , Endorribonucleases/farmacologia , Proteínas Serina-Treonina Quinases , Apoptose , Estresse do Retículo Endoplasmático , RNA Mensageiro
5.
Transfusion ; 64(2): 380-387, 2024 02.
Artigo em Inglês | MEDLINE | ID: mdl-38158881

RESUMO

BACKGROUND: Little s antigen is mainly defined by a single nucleotide polymorphism at c.143C (p.Thr48) on the GYPB gene. Several variants on GYPB can alter the expression of s antigen. The aim of this study was to investigate the molecular basis of variant s antigen expression in the Chinese population. STUDY DESIGN AND METHODS: A total of 4983 whole blood samples were collected to screen the individuals with discrepant s typing results using two different monoclonal anti-s. Then, the sequence of GYPB exon 4 was analyzed by Sanger sequencing. Flow cytometry analysis was performed to quantify s antigen expression on red blood cells (RBCs). In vitro expression study was performed to verify the effect of the GYPB variants identified on the expression of s antigen. RESULTS: Four donors were identified to have discrepant s typing results. Sanger sequencing showed that three donors carried the c.173C > G variant (p.Pro58Arg) specific for sD antigen, the other one carried a novel GYPB (c.160C > T, p.Arg54Cys) variant. Flow cytometry identified a partial and weak expression of s antigen on the RBCs of the four donors. Furthermore, in vitro expression study confirmed the effect of the two variants on the s antigen expression. CONCLUSION: The results demonstrated that in addition to p.Thr48, the two extra amino acids p.Arg54 and p.Pro58 are also important for full expression of s antigen. Since the individuals with partial s antigen are at risk for the development of alloanti-s, it is important to select at least two different monoclonal anti-s for correct s typing.


Assuntos
Antígenos de Grupos Sanguíneos , Glicoforinas , Humanos , Alelos , Glicoforinas/genética , Antígenos de Grupos Sanguíneos/genética , Fenótipo , Eritrócitos/metabolismo , Sistema do Grupo Sanguíneo Rh-Hr/metabolismo
6.
ACS Synth Biol ; 13(1): 54-60, 2024 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-38117980

RESUMO

Transcriptional regulation is of great significance for cells to maintain homeostasis and, meanwhile, represents an innovative but less explored means to control biological processes in synthetic biology and bioengineering. Herein we devised a T7 RNA polymerase (T7RNAP) variant through replacing an essential lysine located in the catalytic core (K631) with Nε-acetyl-l-lysine (AcK) via genetic code expansion. This T7RNAP variant requires the deacetylase activity of NAD-dependent sirtuins to recover its enzymatic activities and thereby sustains sirtuin-dependent transcription of the gene of interest in live cells including bacteria and mammalian cells as well as in in vitro systems. This T7RNAP variant could link gene transcription to sirtuin expression and NAD availability, thus holding promise to support some relevant research.


Assuntos
Sirtuínas , Animais , Sirtuínas/genética , Sirtuínas/metabolismo , NAD/metabolismo , RNA Polimerases Dirigidas por DNA/genética , RNA Polimerases Dirigidas por DNA/metabolismo , Proteínas Virais/genética , Mamíferos/metabolismo
7.
IEEE Trans Image Process ; 32: 6386-6400, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37963006

RESUMO

Action Quality Assessment (AQA) plays an important role in video analysis, which is applied to evaluate the quality of specific actions, i.e., sports activities. However, it is still challenging because there are lots of small action discrepancies with similar backgrounds, but current approaches mostly adopt holistic video representations. So that fine-grained intra-class variations are unable to be captured. To address the aforementioned challenge, we propose a Fine-grained Spatio-temporal Parsing Network (FSPN) which is composed of the intra-sequence action parsing module and spatiotemporal multiscale transformer module to learn fine-grained spatiotemporal sub-action representations for more reliable AQA. The intra-sequence action parsing module performs semantical sub-action parsing by mining sub-actions at fine-grained levels. It enables a correct description of the subtle differences between action sequences. The spatiotemporal multiscale transformer module learns motion-oriented action features and obtains their long-range dependencies among sub-actions at different scales. Furthermore, we design a group contrastive loss to train the model and learn more discriminative feature representations for sub-actions without explicit supervision. We exhaustively evaluate our proposed approach in the FineDiving, AQA-7, and MTL-AQA datasets. Extensive experiment results demonstrate the effectiveness and feasibility of our proposed approach, which outperforms the state-of-the-art methods by a significant margin.

8.
Microorganisms ; 11(9)2023 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-37764124

RESUMO

Human rotavirus (HRV) is an enteric virus that causes infantile diarrhea. However, the risk factors contributing to HRV colonization in young adults have not been thoroughly investigated. Here, we compared the differences in dietary habits and composition of gut microbiota between asymptomatic HRV-infected young adults and their healthy counterparts and investigated potential risk factors contributing to HRV colonization. Our results indicated that asymptomatic HRV-infected adults had an excessive intake of milk and dairy and high levels of veterinary antibiotics (VAs) and preferred veterinary antibiotic (PVAs) residues in urine samples. Their gut microbiota is characterized by abundant Gram-negative (G-) bacteria and high concentrations of lipopolysaccharide (LPS). Several opportunistic pathogens provide discriminatory power to asymptomatic, HRV-infected adults. Finally, we observed an association between HRV colonization and disrupted gut microbiota caused by the exposure to VAs and PVAs. Our study reveals the traits of disrupted gut microbiota in asymptomatic HRV-infected adults and provides a potential avenue for gut microbiota-based prevention strategies for HRV colonization.

9.
Angew Chem Int Ed Engl ; 62(40): e202308472, 2023 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-37587083

RESUMO

Genetically replacing an essential residue with the corresponding photocaged analogues via genetic code expansion (GCE) constitutes a useful and unique strategy to directly and effectively generate photoactivatable proteins. However, the application of this strategy is severely hampered by the limited number of encoded photocaged proteinogenic amino acids. Herein, we report the genetic incorporation of photocaged glutamic acid analogues in E. coli and mammalian cells and demonstrate their use in constructing photoactivatable variants of various fluorescent proteins and SpyCatcher. We believe genetically encoded photocaged Glu would significantly promote the design and application of photoactivatable proteins in many areas.


Assuntos
Escherichia coli , Ácido Glutâmico , Animais , Ácido Glutâmico/genética , Escherichia coli/genética , Proteínas/química , Aminoácidos , Código Genético , Mamíferos
10.
Ecol Evol ; 13(5): e10047, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-37139404

RESUMO

Earthworms modulate carbon and nitrogen cycling in terrestrial ecosystems, but their effect may be compromised by the deposition of pollutants from industrial emissions. However, studies investigating how deposited compounds affect the role of earthworms in carbon cycling such as litter decomposition are lacking, although the interactions of earthworms and deposited compounds are important for understanding the impact of pollutants on ecosystems and the potential of earthworms in bioremediation. We performed a 365-day in situ litterbag decomposition experiment in a deciduous (Quercus variabilis) and coniferous (Pinus massoniana) forest in southeast China. We manipulated nitrogen (N), sodium (Na), and polycyclic aromatic hydrocarbons (PAHs) as model compounds during litter decomposition with and without earthworms (Eisenia fetida). After one year, N, Na, and PAH all slowed down litter mass loss, with the effects of Na being the strongest. By contrast, E. fetida generally increased litter mass loss, and the positive effects were uniformly maintained irrespective of the type of compounds added. However, the pathways to how earthworms increased litter mass loss varied among the compounds added and the two forests studied. As indicated by structural equation modeling, earthworms mitigated the negative effects of deposited compounds by directly increasing litter mass loss and indirectly increasing soil pH and microbial biomass. Overall, the results indicate that the acceleration of litter mass loss by earthworms is little affected by deposited compounds, and that earthworms have the potential to mitigate negative impacts of pollutants on litter decomposition and ecosystem processes.

11.
Biol Trace Elem Res ; 201(12): 5561-5574, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36964416

RESUMO

To explore the association between serum-related indicators (levels of inflammatory cytokines and essential trace elements) and miscarriage risk among infertile women undergoing assisted reproductive techniques (ART) on the 14th day after embryo transfer, and to develop and establish a multivariable algorithm model that might predict pregnancy outcome. According to a nested case-control study design, a total of 100 miscarriage cases and 100 live birth controls were included in this study, and women in both groups were infertile and have underwent in vitro fertilization (IVF). Pregnancy tests were performed and serum levels of five essential trace elements (vanadium (V), copper (Cu), zinc (Zn), selenium (Se) and molybdenum (Mo)) and five inflammatory cytokines (interleukin-1ß (IL-1ß), IL-6, IL-8, IL-10 and tumor necrosis factor-α (TNF-α)) of the participants were measured on the 14th day after embryo transfer. The serum levels of five inflammatory cytokines were determined by multiple magnetic bead enzyme immunity analyzer; and the serum concentrations of five elements were determined simultaneously by inductively coupled plasma‒mass spectrometry (ICP ‒ MS). The logistic regression was used to evaluate the relationship between these serum indices and miscarriage risk among women undergoing ART, and a predictive model of pregnancy outcome based on these indices was established. The levels of IL-10, IL-1ß and TNF-α of infertile women in the live birth group were significantly higher than those in the miscarriage group (p = 0.009, p < 0.001, p = 0.006), and the levels of V, Cu, Zn and Se of infertile women in the live birth group were also significantly higher than those in the miscarriage group (all p < 0.001). Through logistic regression analyses, we found that serum levels of IL-1ß, TNF-α, V, Cu, Zn and Se were significantly and negatively associated with miscarriage risk. Different combination prediction models were generated according to the results of logistic regression analyses, and the combination of IL-1ß, Cu and Zn had the best prediction performance. The area under the curve (AUC) was 0.776, the sensitivity of the model was 60% and the specificity was 84%. In conclusion, the serum-related indicators of women undergoing ART on the 14th day after embryo transfer, including the inflammatory cytokines such as IL-1ß and TNF-α and the essential trace metal elements such as V, Cu, Zn and Se, were negatively correlated with miscarriage risk. A multivariate algorithm model to predict pregnancy outcome among women undergoing ART was established, which showed that IL-1ß, Cu and Zn might synergistically predict pregnancy outcome.


Assuntos
Aborto Espontâneo , Infertilidade Feminina , Selênio , Oligoelementos , Feminino , Humanos , Gravidez , Aborto Espontâneo/diagnóstico , Aborto Espontâneo/metabolismo , Estudos de Casos e Controles , Fertilização in vitro , Infertilidade Feminina/terapia , Interleucina-10 , Interleucina-1beta/sangue , Interleucina-1beta/metabolismo , Fator de Necrose Tumoral alfa , Zinco/sangue , Cobre/sangue
12.
Environ Pollut ; 325: 121434, 2023 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-36907243

RESUMO

Cadmium (Cd) is a widespread environmental pollutant and a reproductive toxicant. It has been proved that Cd can reduce male fertility, however, the molecular mechanisms remain unveiled. This study aims to explore the effects and mechanisms of pubertal Cd exposure on testicular development and spermatogenesis. The results showed that Cd exposure during puberty could cause pathological damage to testes and reduce sperm counts in mice in adulthood. Moreover, Cd exposure during puberty reduced GSH content, induced iron overload and ROS production in testes, suggesting that Cd exposure during puberty may induce testicular ferroptosis. The results in vitro experiments further strengthened that Cd caused iron overload and oxidative stress, and decreased MMP in GC-1 spg cells. In addition, Cd disturbed intracellular iron homeostasis and peroxidation signal pathway based on transcriptomics analysis. Interestingly, these changes induced by Cd could be partially suppressed by pretreated with ferroptotic inhibitors, Ferrostatin-1 and Deferoxamine mesylate. In conclusion, the study demonstrated that Cd exposure during puberty maybe disrupted intracellular iron metabolism and peroxidation signal pathway, triggered ferroptosis in spermatogonia, and ultimately damaged testicular development and spermatogenesis in mice in adulthood.


Assuntos
Ferroptose , Sobrecarga de Ferro , Masculino , Camundongos , Animais , Cádmio/metabolismo , Maturidade Sexual , Sêmen/metabolismo , Espermatogênese , Testículo , Estresse Oxidativo , Ferro/metabolismo
13.
Toxicology ; 486: 153448, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36731763

RESUMO

Cadmium (Cd) is a toxic metal and also a well-known reproductive toxicant. Cd could induce germ cells apoptosis in mouse testes, however, the mechanism remains unclear. This study designed in vitro using GC-1 spermatogonial (spg) cells to explore the cytotoxicity and the molecular mechanisms induced by cadmium chloride(CdCl2). As expected, CdCl2 elevated the levels of reactive oxygen species (ROS) and induced the release of AIF and Cyt-c from the mitochondria to the cytosol in spermatogonia. Correspondingly, CdCl2 apparently increased the apoptotic rate in spermatogonia. Further researches found that CdCl2 could activate IP3R-MCU pathway, trigger Ca2+ transfer from endoplasmic reticulum to mitochondria, and cause mitochondrial Ca2+ overload. BAPTA acetoxymethyl ester (BAPTA-AM), a calcium chelator, almost completely attenuated IP3R phosphorylation, inhibited the mRNA and protein expression levels of VDAC1, MCU and MCUR1 upregulated by CdCl2, reduced the calcium ion content in the mitochondria. Moreover, BAPTA-AM could decrease the level of ROS, antagonize CdCl2-induced release of AIF and Cyt-c from the mitochondria to the cytosol and alleviate CdCl2-induced apoptosis in spermatogonia. As above, these results provided the evidence that CdCl2 might induce apoptosis of spermatogonia via mitochondrial Ca2+ overload mediated by IP3R-MCU signal pathway.


Assuntos
Cádmio , Cálcio , Masculino , Camundongos , Animais , Cádmio/toxicidade , Cálcio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Espermatogônias/metabolismo , Mitocôndrias/metabolismo , Transdução de Sinais , Apoptose
14.
Transfusion ; 63(5): 912-917, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36852717

RESUMO

BACKGROUND: Mutation in the FUT1 gene can impact the structure and function of α-(1,2)-fucosyltransferase 1 (α2FucT1). To explain the para-Bombay phenotype of a novel FUT1 allele, three-dimensional (3D) modeling and mutation effect analysis of α2FucT1 were performed by bioinformatic tools. MATERIALS AND METHODS: Blood and saliva samples were collected from a patient who was suspected to be a para-Bombay phenotype. H, A, and B antigens were determined with routine serologic methods for those samples. FUT1 and FUT2 coding regions were determined by Sanger sequencing. The novel heterozygous mutation was confirmed by cloning and sequencing. 3D model of mutant α2FucT1 was built by Phyre 2 and the mutation effect was evaluated by Chimera, PROVEAN, and Polyphen-2. RESULTS: Weak H, A, and B antigens were detected on RBCs of the proband and normal quantities of H, A, and B antigens were observed in his saliva. Cloning sequencing showed that the proband carried a novel FUT1 allele (c.889C>T, p.Leu297Phe) and a null FUT1*01N.06 allele. 3D model showed that the p.Leu297Phe variant in α2FucT1 reduced the number of hydrogen bonds and the mutation effect was predicted to be deleterious and possibly damaging, which suggested that the conformation and activity of the enzyme might be significantly damaged. CONCLUSION: A novel missense mutation led to an amino acid variant p.Leu297Phe in α2FucT1, which was a potential cause of the inactivation of the enzyme. Computational evaluation was a convenient and useful approach for the mutation effect analysis of the enzyme.


Assuntos
Mutação de Sentido Incorreto , Alelos , Genótipo , Mutação , Fenótipo , Humanos
15.
Heliyon ; 9(1): e12718, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36685479

RESUMO

Objective: Rotavirus (RV), one of non-enveloped double-strained RNA viruses, can cause infantile diarrheal illness. It is widely accepted that RV is transmitted mainly via feces-oral route. However, infected asymptomatic adults are becoming the source of infection. It is necessary to explore the underlying mechanism of RV replication in adult's intestine. Methods: After recruiting healthy volunteers and RV asymptomatic carriers, we firstly investigated the association of animal-derived food intake with antibiotic level in urine samples. Secondly, we compared the difference in the structure of gut microbiota, and identified the taxa that most likely explained the difference. Finally, we investigated the impact of lipopolysaccharide (LPS), produced by gram-negative bacteria, on RV replication in vivo and in vitro. Results: We found that 10% of participants were RV asymptomatic carriers in our study. High intake of animal-derived food was positively correlated to antibiotic level in urine samples. The disrupted gut microbiota in RV carriers was characterized by high abundance of antibiotic resistant gram-negative bacteria and high level of LPS. The disrupted gut microbiota caused by penicillin treatment was benefit to RV replication in vivo. LPS enhanced RV thermal stability in vitro. Conclusions: Our findings suggest that the imbalanced gut microbiota caused by antibiotic exposure plays an important role in RV replication, and brings risk to health complications.

16.
Blood ; 141(17): 2141-2150, 2023 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-36638337

RESUMO

Red blood cells (RBCs) of Asian-type DEL phenotype express few RhD proteins and are typed as serologic RhD-negative (D-) phenotype in routine testing. RhD-positive (D+) RBC transfusion for patients with Asian-type DEL has been proposed but has not been generally adopted because of a lack of direct evidence regarding its safety and the underlying mechanism. We performed a single-arm multicenter clinical trial to document the outcome of D+ RBC transfusion in patients with Asian-type DEL; none of the recipients (0/42; 95% confidence interval, 0-8.40) developed alloanti-D after a median follow-up of 226 days. We conducted a large retrospective study to detect alloanti-D immunization in 4045 serologic D- pregnant women throughout China; alloanti-D was found only in individuals with true D- (2.63%, 79/3009), but not in those with Asian-type DEL (0/1032). We further retrospectively examined 127 serologic D- pregnant women who had developed alloanti-D and found none with Asian-type DEL (0/127). Finally, we analyzed RHD transcripts from Asian-type DEL erythroblasts and examined antigen epitopes expressed by various RHD transcripts in vitro, finding a low abundance of full-length RHD transcripts (0.18% of the total) expressing RhD antigens carrying the entire repertoire of epitopes, which could explain the immune tolerance against D+ RBCs. Our results provide multiple lines of evidence that individuals with Asian-type DEL cannot produce alloanti-D when exposed to D+ RBCs after transfusion or pregnancy. Therefore, we recommend considering D+ RBC transfusion and discontinuing anti-D prophylaxis in patients with Asian-type DEL, including pregnant women. This clinical trial is registered at www.clinicaltrials.gov as #NCT03727230.


Assuntos
Antígenos de Grupos Sanguíneos , Sistema do Grupo Sanguíneo Rh-Hr , Humanos , Feminino , Gravidez , Estudos Retrospectivos , Sistema do Grupo Sanguíneo Rh-Hr/genética , Transfusão de Sangue , Eritrócitos , Fenótipo , Epitopos , Alelos
17.
Microvasc Res ; 147: 104491, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36709858

RESUMO

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) are characterized by pulmonary microvascular endothelial cells (PMVECs) barrier dysfunction and proinflammatory cytokine influx into lung tissue, resulting in pulmonary oedema. Ceramide overproduction is an important mediator of pulmonary hyperinflammation and pulmonary oedema in Acute lung injury (ALI). Ceramides induce NLRP3 inflammasome activation are essential for the hyperinflammatory response. However, the roles and specific mechanisms of ceramide-induced NLRP3 inflammasome activation, proinflammatory cytokine manufacturing and PMVECs barrier dysfunction in ALI are unclear. Herein, pretreatment with the acid sphingomyelinase (ASMase) inhibitor imipramine (but not a neutral sphingomyelinase (NSMase) inhibitor or de novo pathway inhibitor) significantly inhibited ceramide early production in rats with lipopolysaccharide (LPS)-induced ALI; Furthermore, the Txnip/NLRP3 inflammasome activation, proinflammatory cytokine release, increased PMVECs permeability and lung injury were significantly decreased. Verapamil, a Txnip inhibitor, substantially inhibited Txnip/NLRP3 inflammasome activation, proinflammatory cytokine release, increased PMVECs permeability and lung injury in rats with C8-ceramide-induced ALI. In vitro, short hairpin RNA-mediated Txnip silencing significantly inhibited C8-ceramide-induced Txnip/NLRP3 inflammasome activation in NR8383 alveolar macrophages (AMs) and early secretion of the proinflammatory cytokines IL-1ß (4-12 h) as well as IL-6 and TNF-α at subsequent times (later than 12 h). However, C8-ceramide significantly increased the early secretion (within 8 h) of the proinflammatory cytokines IL-1ß, IL-6 and TNF-α in a co-culture model of NR8383 AMs and PMVECs, and Txnip silencing of NR8383 AMs inhibited the secretion of pro-inflammatory cytokines and reduced cytoskeletal rearrangements, intercellular connection breakage and hyperpermeability in PMVECs. Overall, our results suggest that in LPS-induced ALI, ceramide-mediated Txnip/NLRP3 inflammasome activation in NR8383 AMs leads to early IL-1ß release, subsequently inducing PMVECs injury and release of the proinflammatory cytokines IL-6 and TNF-α, ultimately leading to PMVECs barrier dysfunction and ALI.


Assuntos
Lesão Pulmonar Aguda , Edema Pulmonar , Ratos , Animais , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Lipopolissacarídeos , Esfingomielina Fosfodiesterase/efeitos adversos , Células Endoteliais/metabolismo , Fator de Necrose Tumoral alfa , Ceramidas/efeitos adversos , Interleucina-6 , Citocinas/metabolismo , Lesão Pulmonar Aguda/metabolismo , Proteínas de Ciclo Celular
18.
Transfusion ; 63(2): 402-414, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36382965

RESUMO

BACKGROUND: The molecular basis of the D variant phenotype in the Chinese differs greatly from that of the Caucasian. Adapting a specific D typing strategy to the spectrum of prevalent RHD variant alleles is necessary. STUDY DESIGN AND METHODS: Blood samples with ambiguous D phenotypes were collected in the Southern Chinese population. A special three-step typing strategy was applied. First, the common DVI type 3 was identified from epitope profiles of D antigen. Then, another common weak D type 15 (RHD*845A) was identified by epitope profiles of D antigen and Sanger sequencing of RHD exon 6. Finally, the remaining D variants were genotyped mainly by Sanger sequencing. For the novel RHD alleles in the coding region and exon-intron junction, in vitro transfection and minigene splicing assays were performed, respectively. The anti-D investigation was performed. RESULTS: DVI type 3 (65/253, 25.7%) and weak D type 15 (62/253, 24.5%) were common Chinese D variants, and RHD*960A, DFR, RHD*weak D type 25, 72, and 136 were frequent variant RHD alleles. Besides, twenty-two sporadic and seven novel RHD alleles (RHD*188A; RHD*688C; RHD*782 T; RHD*1181C; RHD*165 T, 993A; RHD*148 + 3G > T and RHD*1227 + 5G > C) were identified. The deleterious effect of the novel RHD alleles on D antigen or mRNA expression was confirmed. Anti-D was detected in two DVI type 3 pregnant women. DISCUSSION: The three-step typing strategy provides an effective approach for Chinese D variant typing. It can be anticipated that commercially available RHD genotyping kits have limitations for testing Chinese D variants, as some of the frequent variants are not interrogated.


Assuntos
População do Leste Asiático , Sistema do Grupo Sanguíneo Rh-Hr , Gravidez , Feminino , Humanos , Alelos , Genótipo , Fenótipo , Epitopos , Sistema do Grupo Sanguíneo Rh-Hr/genética
19.
J Fungi (Basel) ; 8(9)2022 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-36135625

RESUMO

Solar radiation has been regarded as a driver of litter decomposition in arid and semiarid ecosystems. Photodegradation of litter organic carbon (C) depends on chemical composition and water availability. However, the chemical changes in organic C that respond to solar radiation interacting with water pulses remain unknown. To explain changes in the chemical components of litter organic C exposed to UV-B, UV-A, and photosynthetically active radiation (PAR) mediated by water pulses, we measured the chemistry of marcescent Lindera glauca leaf litter by solid-state 13C cross-polarization magic angle spinning (CPMAS) nuclear magnetic resonance (NMR) over 494 days of litter decomposition with a microcosm experiment. Abiotic and biotic factors regulated litter decomposition via three pathways: first, photochemical mineralization of lignin methoxyl C rather than aromatic C exposed to UV radiation; second, the biological oxidation and leaching of cellulose O-alkyl C exposed to PAR and UV radiation interacts with water pulses; and third, the photopriming effect of UV radiation on lignin aromatic C rather than cellulose O-alkyl C under the interaction between radiation and water pulses. The robust decomposition index that explained the changes in the mass loss was the ratio of aromatic C to O-alkyl C (AR/OA) under radiation, but the ratio of hydrophobic to hydrophilic C (hydrophobicity), the carbohydrate C to methoxyl C ratio (CC/MC), and the alkyl C to O-alkyl C ratio (A/OA) under radiation were mediated by water pulses. Moreover, the photopriming effect and water availability promoted the potential activities of peroxidase and phenol oxidase associated with lignin degradation secreted by fungi. Our results suggest that direct photodegradation of lignin methoxyl C increases microbial accessibility to lignin aromatic C. Photo-oxidized compounds might be an additional C pool to regulate the stability of the soil C pool derived from plant litter by degrading lignin methoxyl and aromatic C.

20.
Front Surg ; 9: 956431, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36117818

RESUMO

Background and purpose: With the clinical application of minimally invasive surgery and concept of enhanced recovery after surgery, the incidence of postoperative complications in lung cancer patients has been significantly reduced. However, postoperative subcutaneous emphysema (SE) becomes the main factor affecting the early discharge of patients. The aim of this study was to analyze the clinical outcome and risk factors for postoperative SE in lung cancer patients. Methods: The clinical data of 414 lung cancer patients who were admitted to the Department of Thoracic Surgery, West China Hospital, Sichuan University from September 2021 to December 2021 were prospectively collected. The incidence, severity and treatment of patients who had SE, surgery approach, application of drainage tube and clinical information were analyzed. Results: The incidence rate of postoperative SE in patients with lung cancer was 33.09% (137/414) and mild cases accounted for the vast majority (30.19%, 125/414). Multivariate analysis indicated that male [odds ratio (OR) = 2.247, P = .014] and advanced age (OR = 1.021, P = .043) were main risk factors for postoperative SE in patients with lung cancer. Conservative treatment was the main treatment option for SE (98.5%, 135/137). The average hospital stay in the subcutaneous emphysema group (5.49 ± 4.41 days) was significantly longer than that in the non-subcutaneous emphysema group (4.44 ± 3.32 days) (P = .014) and no significant statistical difference in the average total hospital cost between the two groups (7,798.31 ± 1,414.85$ vs. 7,501.14 ± 1,605.18$, P = .072). Conclusion: Postoperative SE in patients with minimally invasive lung cancer is mainly mild, and conservative treatment is appropriate for most cases.

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