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Cytokine release syndrome (CRS) is a great challenge for the application of anti-CD19 CAR-T cell therapy. The aim of this study was to investigate the effect of knocking down interferon gamma (IFN-γ) by shRNA as a potential strategy to reduce the cytokine storms. A newly designed short hairpin interference RNA of IFN-γ (shIFN-γ) in CD19CAR gene was constructed. Several cellular model systems of approach using Nalm-6 cell lines including Nalm-6CD19pos and Nalm-6CD19neg with or without monocytes and endothelial cells were used to analyze the different levels of cytokines after shIFN-γ-anti-CD19CAR-T cell targeted therapy. The activity of this novel CD19CAR-T was evaluated both in vitro and in NSG mouse model. The killing efficacy of shIFN-γ-anti-CD19CAR-T at the E:T ratio of 2:1 was similar to that of regular anti-CD19CAR-T at the E:T ratio of 1:1. The IFN-γ level in the shIFN-γ-anti-CD19CAR-T cell group was (2673.1 ± 307.4) pg/ml at the E:T ratio of 2:1 which was significantly lower than that ((8261.5 ± 345.5) pg/ml) in the regular anti-CD19CAR-T group at the E:T ratio of 1:1. Cytotoxicity experiments in vitro showed significantly reduced concentrations of IFN-γ, IL-6 and TNFα in the shIFN-γ-anti-CD19CAR-T cell group compared to regular anti-CD19CAR-T cell group. Both regular anti-CD19CAR and shIFN-γ-CD19CAR-T exerted bystander killing effect in vitro. We conclude that shIFN-γ-anti-CD19CAR-T cells can reduce the generation of cytokine storms without significantly compromising their therapeutic efficacy in the preclinical setting. In mouse model, 3 × 106 shIFN-γ-anti-CD19CAR-T cells/mouse generated the similar killing efficacy to that with 2 × 106 regular anti-CD19CAR-T cells/mouse.
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Citocinas , Interferon gama , Animais , Camundongos , Citocinas/genética , Interferon gama/genética , Síndrome da Liberação de Citocina , Células Endoteliais , ApoptoseRESUMO
BACKGROUND: Guillain-Barré syndrome (GBS), a post-infectious, immune-mediated, acute demyelinating disease of the peripheral nerves and nerve roots, represents the most prevalent and severe acute paralyzing neuropathy. Purinergic P2X7 receptors (P2X7R) play a crucial role in central nervous system inflammation. However, little is known about their role in the immune-inflammatory response within the peripheral nervous system. METHODS: Initially, we assessed the expression of purinergic P2X7R in the peripheral blood of patients with GBS using flow cytometry and qRT-PCR. Next, we explored the expression of P2 X7R in CD4+ T cells, CD8+ T cells, and macrophages within the sciatic nerves and spleens of rats using immunofluorescence labeling and flow cytometry. The P2X7R antagonist brilliant blue G (BBG) was employed to examine its therapeutic impact on rats with experimental autoimmune neuritis (EAN) induced by immunization with the P0180 - 199 peptide. We analyzed CD4+ T cell differentiation in splenic mononuclear cells using flow cytometry, assessed Th17 cell differentiation in the sciatic nerve through immunofluorescence staining, and examined the expression of pro-inflammatory cytokine mRNA using RT-PCR. Additionally, we performed protein blotting to assess the expression of P2X7R and NLRP3-related inflammatory proteins within the sciatic nerve. Lastly, we utilized flow cytometry and immunofluorescence labeling to examine the expression of NLRP3 on CD4+ T cells in rats with EAN. RESULTS: P2X7R expression was elevated not only in the peripheral blood of patients with GBS but also in rats with EAN. In rats with EAN, inhibiting P2X7R with BBG alleviated neurological symptoms, reduced demyelination, decreased inflammatory cell infiltration of the peripheral nerves, and improved nerve conduction. BBG also limited the production of pro-inflammatory molecules, down-regulated the expression of P2X7R and NLRP3, and suppressed the differentiation of Th1 and Th17 cells, thus protecting against EAN. These effects collectively contribute to modifying the inflammatory environment and enhancing outcomes in EAN rats. CONCLUSIONS: Suppression of P2X7R relieved EAN manifestation by regulating CD4+ T cell differentiation and NLRP3 inflammasome activation. This finding underscores the potential significance of P2X7R as a target for anti-inflammatory treatments, advancing research and management of GBS.
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Síndrome de Guillain-Barré , Neurite Autoimune Experimental , Antagonistas do Receptor Purinérgico P2X , Animais , Humanos , Ratos , Linfócitos T CD8-Positivos , Diferenciação Celular/efeitos dos fármacos , Síndrome de Guillain-Barré/tratamento farmacológico , Inflamassomos/efeitos dos fármacos , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Antagonistas do Receptor Purinérgico P2X/farmacologia , Antagonistas do Receptor Purinérgico P2X/uso terapêutico , Nervo Isquiático/metabolismo , Células Th17/efeitos dos fármacos , Células Th17/metabolismo , Células Th1/efeitos dos fármacos , Células Th1/metabolismoRESUMO
BACKGROUND: Cellular immunotherapy, represented by the chimeric antigen receptor T cell (CAR-T), has exhibited high response rates, durable remission, and safety in vitro and in clinical trials. Unfortunately, anti-CD19 CAR-T (CART-19) treatment alone is prone to relapse and has a particularly poor prognosis in relapsed/refractory (r/r) B-ALL patients. To date, addressing or reducing relapse remains one of the research priorities to achieve broad clinical application. METHODS: We manufactured second generation CART-19 cells and validated their efficacy and safety in vitro and in vivo. Through co-culture of Nalm-6 cells with short-term cultured CART-19 cells, CD19-negative Nalm-6 cells were detected by flow cytometry, and further investigation of the relapsed cells and their resistance mechanisms was evaluated in vitro. RESULTS: In this study, we demonstrated that CART-19 cells had enhanced and specific antileukemic activities, and the survival of B-ALL mouse models after CART-19 treatment was significantly prolonged. We then shortened the culture time and applied the serum-free culture to expand CAR-T cells, followed by co-culturing CART-19 cells with Nalm-6 cells. Surprisingly, we observed the proliferation of CD19-negative Nalm-6 cells around 28 days. Identification of potential resistance mechanisms showed that the relapsed cells express truncated CD19 proteins with decreased levels and, more importantly, CAR expression was detected on the relapsed cell surface, which may ultimately keep them antigen-negative. Furthermore, it was validated that CART-22 and tandem CART-22/19 cells could effectively kill the relapsed cells, but neither could completely eradicate them. CONCLUSIONS: We successfully generated CART-19 cells and obtained a CD19-negative refractory relapsed B-ALL cell line, providing new insights into the underlying mechanisms of resistance and a new in vitro model for the treatment of r/r B-ALL patients with low antigen density.
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Antígenos CD19 , Receptores de Antígenos Quiméricos , Antígenos CD19/metabolismo , Antígenos CD19/imunologia , Animais , Humanos , Receptores de Antígenos Quiméricos/metabolismo , Receptores de Antígenos Quiméricos/imunologia , Linhagem Celular Tumoral , Imunoterapia Adotiva/métodos , Resistencia a Medicamentos Antineoplásicos , Camundongos , Técnicas de Cocultura , Ensaios Antitumorais Modelo de Xenoenxerto , Leucemia-Linfoma Linfoblástico de Células Precursoras B/terapia , Leucemia-Linfoma Linfoblástico de Células Precursoras B/imunologiaRESUMO
Hantaan virus (HTNV) infection can cause hemorrhagic fever with renal syndrome (HFRS) in humans, and currently, there are no long-standing protective vaccines or specific antivirals available. Guanylate-binding protein 1 (GBP1) is an interferon-stimulated gene that defends against various pathogen infections. However, the function of GBP1 in HTNV infection remains unknown. Here, we describe how GBP1 prevents HTNV infection by obstructing virus entry. We found that HTNV infection induced GBP1 expression and that overexpression of GBP1 inhibited HTNV infection, while knockout of GBP1 had the opposite effect. Interestingly, GBP1 did not affect interferon (IFN) signaling during HTNV infection. Instead, GBP1 prevented HTNV from entering cells through clathrin-mediated endocytosis (CME). We also discovered that GBP1 specifically interacted with actin but not dynamin 2 (DNM2) and made it difficult for DNM2 to be recruited by actin, which may account for the suppression of CME during HTNV infection. These findings establish an antiviral role for GBP1 in inhibiting HTNV infection and help us better understand how GBP1 regulates HTNV entry and could potentially aid in developing treatments for this virus.
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Endocitose , Proteínas de Ligação ao GTP , Vírus Hantaan , Internalização do Vírus , Humanos , Actinas/metabolismo , Linhagem Celular , Dinamina II/metabolismo , Dinamina II/genética , Proteínas de Ligação ao GTP/genética , Proteínas de Ligação ao GTP/metabolismo , Vírus Hantaan/fisiologia , Células HEK293 , Febre Hemorrágica com Síndrome Renal/virologia , Interações Hospedeiro-PatógenoRESUMO
Eltrombopag combined with immunosuppressive therapy (IST) was superior to IST alone for severe aplastic anemia (SAA) in the previous studies. But in China, horse antithymocyte globulin (hATG) is not available, instead, we use rabbit ATG (rATG). Here, we compared the efficacy and safety of IST (rATG combined with cyclosporine) combined with or without eltrombopag for the first-line treatment of SAA and very severe aplastic anemia (VSAA). A total of 371 patients in ten institutions in China from April 1, 2017 to December 1, 2022 were enrolled. The overall response (OR) rate at 3 months (54.2% vs. 41%; P = 0.046), the complete response (CR) (31.3% vs. 19.4%; P = 0.041) and OR (78.3% vs. 51.1%; P < 0.0001) rates at 6 months were significantly higher with IST combined with eltrombopag than with IST alone in SAA patients. While in VSAA patients, the addition of eltrombopag to IST only increased the CR rate at 6 months (29.8% vs. 9.43%; P = 0.010). Liver injury increased significantly in groups treated with IST combined with eltrombopag (P < 0.05). Serious treatment-related toxicities were similar (P > 0.05). In patients with SAA, 3-year failure-free survival (FFS) of eltrombopag combined with IST group was significantly higher than that of IST group (70.7 ± 5.3% vs. 50.3 ± 3.9%; P = 0.007). In patients with VSAA, the addition of eltrombopag significantly improved 3-year overall survival (OS) (82.2 ± 5.7% vs. 57.3 ± 7.2%; P = 0.020). Our findings suggested that IST combined with eltrombopag could improve the hematological recovery of newly diagnosed SAA without increasing severe toxicities. But in VSAA, the addition of eltrombopag seemed to show no other improvement to efficacy except the CR rate at 6 months.
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The synthesis of compounds based on fragments derived from natural products (NPs) serves as a source of inspiration for the design of pseudo-natural products (PNPs), to identify bioactive molecules that exhibit similar characteristics to NPs. These novel molecular scaffolds exhibit previously unexplored biological activities as well. This study reports the development and synthesis of a novel pentacyclic ring system, the indole-pyrimidine-quinoline (IPQ) scaffold. The design of this scaffold was based on the structural characteristics of four natural products, namely tryptanthrin, luotonin A, rutaecarpine, and camptothecin. Several successive steps accomplished the effective synthesis of the IPQ scaffold. The constituent components of the pentacycle, containing the indole, quinazolinone, pyrimidone, and quinoline units, possess significant biological significance. Compound 1a demonstrated noteworthy anti-tumor activity efficacy against A549 cell lines among the tested compounds. The compound 1a was observed to elicit cell cycle arrest in both the G2/M and S phases, as well as trigger apoptosis in A549 cells. These effects were attributed to its ability to modulate the activation of mitochondrial-related mitogen-activated protein kinase (MAPK) signaling pathways.
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Antineoplásicos , Produtos Biológicos , Quinolinas , Antineoplásicos/química , Apoptose , Produtos Biológicos/farmacologia , Produtos Biológicos/química , Camptotecina/farmacologia , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Quinolinas/farmacologia , Indóis/química , Indóis/farmacologia , PirimidinasRESUMO
Cu-SSZ-39 zeolite with 8-membered rings is regarded as a very promising catalyst in the NH3-SCR reaction, but its hydrothermal stability still remains to be improved. One of the solutions to promote hydrothermal stability is the insertion of rare earth elements in the product. Nevertheless, normal ion exchange of rare earth elements limits their contents in the zeolite product due to their large hydrated ionic radius and alkaline environment under hydrothermal conditions. Herein, we for the first time present a new method for the one-pot synthesis of Ce-SSZ-39 zeolite under solvent-free conditions. The key to success is the use of Ce-FAU zeolite as a precursor. The obtained product shows good crystallinity, sheet-like morphology, large BET surface area, and 4-coordinated Al species. Detailed investigations illustrate that Ce species in the Cu/Ce-SSZ-39 zeolite micropore can prevent the dealumination and thus formation of CuAlOx species during hydrothermal aging at 850 °C for 16 h, giving the excellent hydrothermal stability and thus showing the excellent catalytic performance in the NH3-SCR reaction. One-pot synthesis of Ce-SSZ-39 zeolite with excellent catalytic performance might open a new door for developing very efficient selective catalytic reduction (SCR) catalysts in near future.
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Due to the high abundance in the Earth's crust and industrial application, fluoride is widely present in our living environment. However, excessive fluoride exposure causes toxicity in different organs. As the most important detoxification and excretion organ, liver is more easily involved in fluoride toxicity than other organs, and oxidative stress is considered as the key mechanism related with fluoride hepatotoxicity. In this study, we mainly investigated the role of nuclear factor erythroid-derived 2-like 2 (NRF2, a core transcription factor in oxidative stress) in fluoride exposure-induced hepatotoxicity as well as the related mechanism. Herein, liver cells (BNL CL.2) were treated with fluoride in different concentrations. The hepatotoxicity and NRF2 signaling pathway were analyzed respectively. Our results indicated that excessive fluoride (over 1 mM) resulted in obvious toxicity in hepatocyte and activated NRF2 and NRF2 target genes. The increased ROS generation after fluoride exposure suppressed KEAP1-induced NRF2 ubiquitylation and degradation. Meanwhile, fluoride exposure also led to blockage of autophagic flux and upregulation of p62, which contributed to activation of NRF2 via competitive binding with KEAP1. Both pharmaceutical activation and genetic activation of NRF2 accelerated fluoride exposure-induced hepatotoxicity. Thus, the upregulation of NRF2 in hepatocyte after fluoride exposure can be regarded as a cellular self-defense, and NRF2-KEAP1 system could be a novel molecular target against fluoride exposure-induced hepatotoxicity.
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Doença Hepática Induzida por Substâncias e Drogas , Fluoretos , Humanos , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Fluoretos/toxicidade , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Transdução de Sinais/genética , Hepatócitos/metabolismo , Estresse Oxidativo/fisiologia , Autofagia/genéticaRESUMO
One previously undescribed alkaloid, named penifuranone A (1), and three known compounds (2-4) were isolated from the mangrove endophytic fungus Penicillium crustosum SCNU-F0006. The structure of the new alkaloid (1) was elucidated based on extensive spectroscopic data analysis and single-crystal X-ray diffraction analysis. Four natural isolates and one new synthetic derivative of penifuranone A, compound 1a, were screened for their antimicrobial, antioxidant, and anti-inflammatory activities. Bioassays revealed that penifuranone A (1) exhibited strong anti-inflammatory activity in vitro by inhibiting nitric oxide (NO) production in lipopolysaccharide-activated RAW264.7 cells with an IC50 value of 42.2 µM. The docking study revealed that compound 1 exhibited an ideal fit within the active site of the murine inducible nitric oxide synthase (iNOS), establishing characteristic hydrogen bonds.
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Alcaloides , Óxido Nítrico , Penicillium , Penicillium/química , Penicillium/metabolismo , Camundongos , Animais , Alcaloides/química , Alcaloides/farmacologia , Alcaloides/isolamento & purificação , Células RAW 264.7 , Óxido Nítrico/metabolismo , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Óxido Nítrico Sintase Tipo II/metabolismo , Simulação de Acoplamento Molecular , Lipopolissacarídeos , Antioxidantes/farmacologia , Antioxidantes/química , Estrutura MolecularRESUMO
PURPOSE: Ultrasound view of the interlaminar structure is likely to be associated with difficult spinal anesthesia (DSA), and a poor ultrasound view which cannot show the anterior and posterior complex predicts a difficult spinal technique. As our target site is the posterior complex, this study aimed to assess whether the ratio of posterior complex length to depth measured by ultrasound can predict DSA in cesarean delivery. METHODS: Four anesthesiologists with 1-2 years of experience located and marked the puncture interspace using a traditional surface landmark. Subsequently, the ultrasound examiner located and measured the marked interspace via an oblique parasagittal ultrasound scan. The anesthesiologists, who were blinded to the ultrasound results, performed spinal anesthesia using a 25-gauge Whitacre spinal needle. The total number of attempts, including skin punctures and needle passes, was recorded and the DSA was defined as 10 unsuccessful attempts. A multivariable logistic regression analysis was used to determine the independent predictors, and receiver operating characteristic curves were constructed to evaluate the performance of the ratio of posterior complex length to depth for predicting DSA. RESULTS: A total of 397 cesarean delivery parturients with successfully measured posterior complex were included in the analysis. DSA occurred in 64 parturients (16.1%). Reduced length [odds ratio (OR) = 0.010, 95% confidence interval (CI), 0.002-0.062, P < 0.001] and increased depth [OR = 6.127, 95% CI, 2.671-14.056, P < 0.001] of the posterior complex were independently predictive of DSA compared with body mass index, abdominal circumference, and palpable surface landmarks. The ratio of posterior complex length to depth for predicting DSA had an area under the curve of 0.86 (95% CI, 0.82-0.90). The optimal cutoff was 0.23, with a sensitivity of 86% (95% CI, 74-93%) and specificity of 72% (95% CI, 67-77%). CONCLUSION: The ratio of posterior complex length to depth measured by ultrasound demonstrated a considerable accuracy in predicting DSA for inexperienced anesthesiologists. A higher ratio at ultrasound is an indication to evaluate the optimal puncture body position and interspace in the clinic practice. CLINICAL TRIAL REGISTRATION: ChiCTR2200065171 https://www.chictr.org.cn/showproj.html?proj=180855.
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BACKGROUND: This study aimed to analyse the expression of microRNA-223 (miR-223) in embryo culture medium and its correlation with pregnancy outcomes. METHODS: Two hundred and two patients undergoing in vitro fertilisation/intracytoplasmic sperm injection (IVF/ICSI) were divided into clinical pregnancy group (n = 101) and non-pregnant group (n = 101). The baseline data, clinical indicators, and the expression level of miR-223 in the embryo medium were compared between the two groups. Logistic regression analysis was used to analyse the relationship between each index and the pregnancy outcome. Receiver operator characteristic curve was carried out to evaluate the differential ability of miR-223 in pregnancy status. Bioinformatics methods were used to identify the target genes of miR-223 and elucidate their functions. RESULTS: Compared with pregnancy group, the non-pregnancy group exhibited a reduction in miR-223 expression (p < 0.001). Multivariate analysis revealed that miR-223 reduction was an independent factor for pregnancy failure (p < 0.05). The ROC curve demonstrated the discriminative capability of miR-223 in distinguishing pregnancy and non-pregnancy. In addition, bioinformatics analysis indicated that the target genes of miR-223 were predominantly located in the endocytic vesicle membrane and were primarily enriched in adenosine monophosphate-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) signalling pathways. CONCLUSION: In this study, levels of miR-223 in the embryo culture medium predicted pregnancy outcomes in subjects undergoing IVF/ICSI. Low expression of miR-223 was a risk factor for adverse pregnancy outcomes in subjects.
In this study, 202 patients who underwent IVF/ICSI were retrospectively analysed and categorised into pregnant and non-pregnant groups based on their pregnancy status. The examination of embryo culture medium samples from both groups revealed that the non-pregnant group exhibited lower miR-223 expression compared to the pregnant group. Subsequent ROC analysis demonstrated the clinical relevance of miR-223 in effectively distinguishing between pregnant and non-pregnant states. Multi-factor analysis further established that the diminished expression of miR-223 independently influenced the likelihood of successful pregnancy.
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Fertilização in vitro , MicroRNAs , Resultado da Gravidez , Injeções de Esperma Intracitoplásmicas , Humanos , Feminino , Gravidez , MicroRNAs/genética , MicroRNAs/metabolismo , Adulto , Fertilização in vitro/métodos , Prognóstico , Curva ROC , Técnicas de Cultura EmbrionáriaRESUMO
Target-negative relapse after CD19 chimeric antigen receptor engineered (CAR) T cell therapy for patients with B lineage acute lymphoblastic leukemia (B-ALL) presents limited treatment options with dismal outcomes. Although CD22-CAR T cells mediate similarly potent antineoplastic effects in patients with CD19dim or even CD19-negative relapse following CD19-directed immunotherapy, a high rate of relapse associated with diminished CD22 cell surface expression has also been observed. Therefore, it is unclear whether any other therapeutic options are available. Mitoxantrone has shown significant antineoplastic activity in patients with relapsed or refractory leukemia over the past decades, and in some cases, the addition of bortezomib to conventional chemotherapeutic agents has demonstrated improved response rates. However, whether this mitoxantrone and bortezomib combination therapy is effective for those patients who have relapsed B-ALL after receiving CD19-CAR T cell therapy remains to be elucidated. In this study, we established a cellular model system using a CD19-positive B-ALL cell line Nalm-6 to investigate the treatment options for CD19-negative relapsed B-ALL after CD19-CAR T cell therapy. In addition to CD22-CAR T therapy, we observed that the combination of bortezomib and mitoxantrone exhibited effective anti-leukemia activity in the CD19-negative Nalm-6 cell line by downregulating p-AKT and p-mTOR. These results suggest that this combination therapy is a possible option for target-negative refractory leukemia cells after CAR-T cell treatment.
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Antineoplásicos , Leucemia-Linfoma Linfoblástico de Células Precursoras , Receptores de Antígenos Quiméricos , Humanos , Imunoterapia Adotiva/efeitos adversos , Imunoterapia Adotiva/métodos , Mitoxantrona/farmacologia , Mitoxantrona/uso terapêutico , Bortezomib/farmacologia , Bortezomib/uso terapêutico , Apoptose , Recidiva , Linhagem Celular , Antígenos CD19 , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Lectina 2 Semelhante a Ig de Ligação ao Ácido SiálicoRESUMO
Stimulator of interferon genes (STING) serve as an endoplasmic reticulum (ER) protein and modulates innate immune responses to viral contagion. Most investigations involving teleost STING antiviral immunity have examined DNA viruses. Therefore, fish STING signaling events against RNA viruses require additional exploration. Here, common carp STING (named CcSTING) was cloned and characterized. The bioinformatics analyses of CcSTING showed evolutionary conservations and were most closely related to other cyprinid STINGs. Immunofluorescence staining discovered that the CcSTING was chiefly placed in the cytoplasm, specifically within the ER. CcSTING was ubiquitously generated in all analyzed organs, with especially strong expression in the gills and head kidney. Spring viremia of carp virus (SVCV) stimulation and poly(I:C) infection induced the generation of CcSTING in immune-associated organs, as well as in peripheral blood leukocytes. Additional investigations revealed that CcSTING overexpression strongly suppressed SVCV replication in EPC cells. Mechanistically, CcSTING enhanced IFN-1 and ISGs expression following SVCV infection. CcSTING also substantially increased both IFN and NF-κB promoter luciferase activity via a dosage-dependent fashion. Lastly, CcSTING significantly up-regulated both TBK1 and p65 phosphorylation. Collectively, these findings demonstrated the critical role and underlying mechanism of fish STING in response to RNA virus.
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Carpas , Doenças dos Peixes , Vírus de RNA , Infecções por Rhabdoviridae , Rhabdoviridae , Animais , Viremia , Carpas/genética , Carpas/metabolismo , Rhabdoviridae/fisiologia , Proteínas de PeixesRESUMO
BACKGROUND: Stroke is the second leading cause of disease-related death and the third leading cause of disability worldwide. However, how to accurately warn of stroke onset remains extremely challenging. Recently, phenylacetyl glutamine (PAGln) has been implicated in the onset of stroke, but evidences from cohort studies of onset are lacking, especially in patients with first-onset or recurrent. It is necessary to deeply demonstrate the effectiveness of PAGln level on warning stroke onset. METHODS: One hundred fifteen first onset stroke patients, 33 recurrent stroke patients, and 135 non-stroke controls were included in the analysis. Risk factors associated with stroke attacking were evaluated, and plasma PAGln levels were detected via HPLC-MS based method. LASSO regression, Pearson correlation analysis, and univariate analysis were carried out to demonstrate the associations between PAGln levels and risk factors of stroke. Random forest machine learning algorithm was used to build classification models to achieve the distinction of first-onset stroke patients, recurrent stroke patients, and non-stroke controls, and further demonstrate the contribution of PAGln levels in the distinction of stroke onset. RESULTS: The median level of PAGln in the first-onset stroke group, recurrent stroke group, and non-stroke group was 933 ng/mL, 1014 ng/mL, and 556 ng/mL, respectively. No statistical correlation was found between PAGln level and subject's living habits, eating preferences, and concomitant diseases (hypertension, hyperlipidemia, and diabetes). Stroke severity indicators, mainly age and NIHSS score, were found associate with the PAGln levels. Machine learning classification models confirmed that PAGln levels, as the main contributing variable, could be used to distinguish recurrent stroke patients (but not first-onset stroke patients) from non-stroke controls. CONCLUSION: PAGln may be an effective indicator to monitor the recurrence in stroke patients.
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Hipertensão , Acidente Vascular Cerebral , Humanos , Glutamina , Biomarcadores , Estudos de Coortes , Infarto Cerebral , RecidivaRESUMO
Stimulator of interferon genes (STING) is an endoplasmic reticulum (ER)-associated protein that plays critical roles in innate immunity and pathogenesis of various diseases. To date, teleost STING against viral stimulation has been identified, whereas STING signaling events in fish against bacteria are not well understood. In the present study, the open reading frame (ORF) of STING from Asian swamp eel (Monopterus albus) was cloned (named MaSTING) and its roles in bacterial infection were investigated. Amino acid sequence alignment and phylogenetic analysis revealed that MaSTING had conserved structures with mammalian STING and shared the closest relationship with mandarin fish STING. Subcellular localization analysis showed that MaSTING distributed in the whole cytoplasm and mainly co-localized with ER. Expression pattern analysis found that MaSTING was constitutively expressed in all the examined tissues with the highest expression in the liver and spleen. Post stimulation with bacteria and various PAMPs, the expression of MaSTING was induced at indicated time points in the immune-related organs and isolated peripheral blood leucocytes. Furthermore, the mechanism underlying MaSTING against bacterial infection was further studied. The qPCR analysis showed that MaSTING overexpression promoted 2'3'-cGAMP induced the expression of IFN-1, ISG15, Viperin, Mx, IL-1ß and TNF-α. Western blotting assay suggested that MaSTING significantly enhanced the phosphorylation of TANK-binding kinase 1 (TBK1) and p65. MaSTING also significantly increased the luciferase activity of IFN-1 and NF-κB promoters. Taken together, MaSTING is involved in host defense against bacterial infection by inducing the inflammatory response.
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Infecções Bacterianas , Smegmamorpha , Animais , Regulação da Expressão Gênica , Filogenia , Proteínas de Peixes/química , Imunidade Inata/genética , Peixes/metabolismo , Interferons/metabolismo , Mamíferos/metabolismoRESUMO
Gasdermin family proteins are important effector proteins mediating pyroptosis and play an important role in innate immune response. GSDME can be cleaved by inflammatory Caspases at specific sites, releasing an active form of N-terminal fragment that binds to the plasma membrane to form pores and release cellular contents. Here, two GSDME genes, CcGSDME-like (CcGSDME-L) and CcGSDMEa, were cloned from common carp. The sequence similarity of the two genes were very high and more similar to DrGSDMEa of zebrafish in evolution. The expression levels of CcGSDME-L and CcGSDMEa can respond to the stimulation of Edwardsiella tarda. The results of cytotoxicity assay showed that CcGSDMEs were cleaved by the activation of canonical CcNLRP1 inflammasome, leading to obvious pyroptosis characteristics and increased cytotoxicity. In EPC cells, three CcCaspases responded to intracellular LPS stimulation and induced significantly cytotoxicity. In order to clarify the molecular mechanism of CcGSDME-induced pyroptosis, the N-terminal of CcGSDME-L (CcGSDME-L-NT) was expressed in 293T cells, which showed strong cytotoxicity and obvious pyroptosis characteristics. Fluorescence localization assay showed that the CcGSDME-L-NT was expressed on cell membrane, and CcGSDMEa-NT was located on the cell membrane or some organelle membranes. These findings can enrich the knowledge of CcNLRP1 inflammasome and GSDMEs mediated pyroptosis in common carp, and provide basic data for the prevention and treatment of fish infectious diseases.
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Carpas , Inflamassomos , Animais , Inflamassomos/genética , Piroptose/genética , Carpas/genética , Carpas/metabolismo , Peixe-Zebra/metabolismo , Imunidade Inata/genéticaRESUMO
To investigate the inhibitory mechanism of the KIF3A gene on nasopharyngeal carcinoma (NPC) tumor stem cells. Set up a blank control group (BCG), NPC group, and KIF3A silence (si-KIF3A) group. The BCG cells were nasopharyngeal normal epithelial cell line NP69, without any treatment, and were cultured routinely; The NPC group cells are human NPC cell line CNE2 cells, which are not subjected to any treatment and are cultured routinely; si-KIF3A group cells were cultured in the offspring of human NPC cell line CNE2 infected with Lentivirus knockdown KIF3A gene. CCK8 was used to detect the cell proliferation ability, Western blot and qPCR were used to detect protein and related mRNA expression, while cell migration and invasion were detected using Transwell. The KIF3A protein and mRNA in the NPC group were higher than those in the BCG (P<0.05), while those in the si-KIF3A group cells were lower compared to BCG cells (P<0.05). The cell proliferation, migration and invasion activity in the si-KIF3A group were reduced than those in BCG (P<0.05). Si-KIF3A group cells HIF-1, NF-κB was reduced than that of BCG (P<0.05). The expression level of HIF-1, NF-κB protein in si-KIF3A group cells was reduced compared to BCG cells (P<0.05). Knocking down the KIF3A gene can reduce the vitality of NPC stem cells, and inhibit the malignant phenotype of NPC stem cells, via inhibiting HIF-1 and NF-κB expression.
Assuntos
NF-kappa B , Neoplasias Nasofaríngeas , Humanos , Cinesinas/genética , Carcinoma Nasofaríngeo/genética , Neoplasias Nasofaríngeas/genética , Células-Tronco Neoplásicas , NF-kappa B/genética , RNA Mensageiro/genéticaRESUMO
PURPOSE: The goal of this study is to investigate the social isolation (SI) subtypes of patients with breast cancer (BC) and to explore its influencing factors. METHODS: A sample of 303 BC patients participated in the study from September to December, 2021. Latent profile analysis (LPA) was performed to identify SI clusters based on the three sub-scales of the Chinese version of the Social Anxiety Scale, the Chinese version of the Social Avoidance and Distress Scale, and the Chinese version of the Loneliness Scale. RESULTS: We found that SI can be divided into three categories: high-level (Class 1), middle-level (Class 2), and low-level (Class 3), accounting for 20.46%, 33.00%, and 46.54%, respectively. Compared to Class 3, Class 1, which had the lower average monthly income per family member (RMB) (< 3000: OR = 5.298, P = .021; 3000 ~ 5000: OR = 5.320, P = .018), was more likely to suffer from SI due to occupation (Laborer: OR = 12.023, P = .009). Surgery (OR = 14.138, P < .001; OR = 2.777, P = .020), chemotherapy (OR = 10.224, P = .001; OR = 3.545, P = .001); poorer family functioning (OR = .671, P < .001; OR = .801, P = .002), and lower levels of self-transcendence (OR = .806, P < .001; OR = .911, P < .001) were important influencing factors for SI in Class 1 and Class 2 compared to Class 3. CONCLUSION: SI is classifiably heterogeneous among patients with BC. Strategies that identify characteristics of SI and give targeted intervention focusing on family functioning and improving self-transcendence levels contribute to the prevention of SI among patients with BC.
Assuntos
Neoplasias da Mama , Humanos , Feminino , Pacientes , Comportamento Social , SolidãoRESUMO
BACKGROUND: HPV screening tests may improve cervical cancer risk stratification and better guide decisions about follow-up with colposcopy/biopsy. This study aimed to estimate the risk of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) among women with oncogenic HPV types and evaluate the performance of colposcopy in the diagnosis of histologic CIN2 + at Putuo Hospital, Shanghai, China. METHODS: This cross-sectional survey was conducted from February 2020 to December 2022 among women who were referred to colposcopy. Women with high-risk (HR) HPV-positive, cytology testing and colposcopy-directed biopsy were included. RESULTS: Univariate and multivariate analysis indicated that high-grade colposcopic impression ((OR, 17.61%, 95%CI: 11.54-26.85%) was associated with the highest risk for detecting CIN2+, followed by HSIL + cytology (OR, 6.90%, 95%CI: 3.56-13.37%) and HPV16/18 positive (OR, 2.91%, 95%CI: 2.12-3.99%). Overall, CIN2 + was detected in 14.6% of 2007 women. HPV16/18 had higher CIN2 + risks than other HR-HPV genotypes (30.1% vs. 10.2%, P<0.001). Among women with low-grade cytology, 24.1% had CIN2+, and the risks for HPV16/18 (58.2%) were higher than for other HR-HPV(16.8%). For those with high-grade cytology, there was no significant difference between HPV groups ( 75.0% vs. 72.9%, P > 0.05). The diagnostic performance of colposcopy in diagnosis of CIN2 + by senior and junior colposcopists was comparable. CONCLUSIONS: The results indicated that referral to colposcopy is recommended in managing women with HR-HPV positive, and colposcopic impressions provide key clues for identification CIN2+.
Assuntos
Infecções por Papillomavirus , Lesões Intraepiteliais Escamosas , Displasia do Colo do Útero , Neoplasias do Colo do Útero , Gravidez , Feminino , Humanos , Colposcopia , Papillomavirus Humano 16/genética , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/diagnóstico , Estudos Transversais , Papillomavirus Humano 18/genética , China , Displasia do Colo do Útero/diagnóstico , Lesões Intraepiteliais Escamosas/complicações , Papillomaviridae/genética , Detecção Precoce de Câncer/métodosRESUMO
Staphylococcus aureus is a common pathogen found in cheese whose Staphylococcal enterotoxins (SE) are the main pathogenic factors that cause food poisoning. The objective of this study was to construct two models to evaluate the safety of Kazak cheese products in terms of composition, changes in S. aureus inoculation amount, Aw, fermentation temperature during processing, and growth of S. aureus in the fermentation stage. A total of 66 experiments comprised of five levels of inoculation amount (2.7-4 log CFU/g), five levels of Aw (0.878-0.961), and six levels of fermentation temperature (32-44 °C) were performed to confirm the growth of S. aureus and the presence of SE limit conditions. Two artificial neural networks (ANN) successfully described the relationship between the assayed conditions and the growth kinetic parameters (maximum growth rates and lag times) of the strain. The good fitting accuracy (R2 values were 0.918 and 0.976, respectively) showed that the ANN was appropriate. Experimental results showed fermentation temperature had the greatest influence on the maximum growth rate and lag time, followed by the Aw and inoculation amount. Furthermore, a probability model was built to predict the production of SE by logistic regression and neural network under the assayed conditions, which proved to be concordant in 80.8-83.8% of the cases with the observed probabilities. The maximum total number of colonies predicted by the growth model in all combinations detected with SE exceeded 5 log CFU/g. Within the range of variables, the minimum Aw for predicting SE production was 0.938, and the minimum inoculation amount for predicting SE production was 3.22 log CFU/g. Additionally, as competition between S. aureus and lactic acid bacteria (LAB) occurs in the fermentation stage, higher fermentation temperatures are conducive to the growth of LAB, which can reduce the risk of S. aureus producing SE. This study can help manufacturers to make decisions on the most appropriate production parameters for Kazak cheese products and to prevent S. aureus growth and SE production.