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1.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 42(2): 227-233, 2024 Apr 01.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-38597082

RESUMO

OBJECTIVES: This clinical study aimed to assess the trueness of three intraoral scanners for the recor-ding of the maximal intercuspal position (MIP) to provide a reference for clinical practice. METHODS: Ten participants with good occlusal relationship and healthy temporomandibular joint were recruited. For the control group, facebow transferring procedures were performed, and bite registrations at the MIP were used to transfer maxillary and mandibular casts to a mechanical articulator, which were then scanned with a laboratory scanner to obtain digital cast data. For the experimental groups, three intraoral scanners (Trios 3, Carestream 3600, and Aoralscan 3) were used to obtain digital casts of the participants at the MIP following the scanning workflows endorsed by the corresponding manufacturers. Subsequently, measurement points were marked on the control group's digital casts at the central incisors, canines, and first molars, and corresponding distances between these points on the maxillary and mandibular casts were measured to calculate the sum of measured distances (DA). Distances between measurement points in the incisor (DI), canine (DC), and first molar (DM) regions were also calculated. The control group's maxillary and mandibular digital casts with the added measurement points were aligned with the experimental group's casts, and DA, DI, DC, and DM values of the aligned control casts were determined. Statistical analysis was performed on DA, DI, DC, and DM obtained from both the control and experimental groups to evaluate the trueness of the three intraoral scanners for the recording of MIP. RESULTS: In the control group, DA, DI, DC, and DM values were (39.58±6.40), (13.64±3.58), (14.91±2.85), and (11.03±1.56) mm. The Trios 3 group had values of (38.99±6.60), (13.42±3.66), (14.55±2.87), and (11.03±1.69) mm. The Carestream 3600 group showed values of (38.57±6.36), (13.56±3.68), (14.45±2.85), and (10.55±1.41) mm, while the Aoralscan 3 group had values of (38.16±5.69), (13.03±3.54), (14.23±2.59), and (10.90±1.54) mm. Analysis of variance revealed no statistically significant differences between the experimental and control groups for overall deviation DA (P=0.96), as well as local deviations DI (P=0.98), DC (P=0.96), and DM (P=0.89). CONCLUSIONS: With standardized scanning protocols, the three intraoral scanners demonstrated comparable trueness to traditional methods in recording MIP, fulfilling clinical requirements.


Assuntos
Incisivo , Dente Molar , Humanos , Mandíbula , Maxila , Desenho Assistido por Computador , Imageamento Tridimensional , Técnica de Moldagem Odontológica
2.
Mar Drugs ; 22(4)2024 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-38667761

RESUMO

In order to explore the extraction and activity of macroalge glycolipids, six macroalgae (Bangia fusco-purpurea, Gelidium amansii, Gloiopeltis furcata, Gracilariopsis lemaneiformis, Gracilaria sp. and Pyropia yezoensis) glycolipids were extracted with five different solvents firstly. Considering the yield and glycolipids concentration of extracts, Bangia fusco-purpurea, Gracilaria sp. and Pyropia yezoensis were selected from six species of marine macroalgae as the raw materials for the extraction of glycolipids. The effects of the volume score of methanol, solid-liquid ratio, extraction temperature, extraction time and ultrasonic power on the yield and glycolipids concentration of extracts of the above three macroalgae were analyzed through a series of single-factor experiments. By analyzing the antioxidant activity in vitro, moisture absorption and moisturizing activity, the extraction process of Bangia fusco-purpurea glycolipids was further optimized by response surface method to obtain suitable conditions for glycolipid extraction (solid-liquid ratio of 1:27 g/mL, extraction temperature of 48 °C, extraction time of 98 min and ultrasonic power of 450 W). Bangia fusco-purpurea extracts exhibited a certain scavenging effect on DPPH free radicals, as well as good moisture-absorption and moisture retaining activities. Two glycolipids were isolated from Bangia fusco-purpurea by liquid-liquid extraction, silica gel column chromatography and thin-layer chromatography, and they showed good scavenging activities against DPPH free radicals and total antioxidant capacity. Their scavenging activities against DPPH free radicals were about 60% at 1600 µg/mL, and total antioxidant capacity was better than that of Trolox. Among them, the moisturizing activity of a glycolipid was close to that of sorbierite and sodium alginate. These two glycolipids exhibited big application potential as food humectants and antioxidants.


Assuntos
Antioxidantes , Glicolipídeos , Alga Marinha , Glicolipídeos/química , Glicolipídeos/isolamento & purificação , Glicolipídeos/farmacologia , Antioxidantes/farmacologia , Antioxidantes/química , Antioxidantes/isolamento & purificação , Alga Marinha/química , Rodófitas/química , Solventes/química , Picratos/química
3.
Heliyon ; 10(7): e28405, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38560178

RESUMO

Inactivated coronavirus disease 2019 (COVID-19) vaccines showed impaired immunogenicity in some autoimmune diseases, but it remains unclear in primary biliary cholangitis (PBC). This study aimed to explore the antibody response to the inactivated COVID-19 vaccine in individuals with PBC, as well as to evaluate coverage, safety, and attitudes toward the COVID-19 vaccine among them. Two cohorts of patients with PBC were enrolled in this study. One cohort was arranged to evaluate the immunogenicity of the inactivated COVID-19 vaccine, another cohort participated in an online survey. The titers of the anti-receptor-binding domain (RBD)-specific immunoglobulin G (IgG), neutralizing antibody (NAb) toward severe acute respiratory syndrome coronavirus 2 wild-type, and NAb toward Omicron BA.4/5 subvariants were detected to assess antibody response from the vaccine. After booster vaccination for more than six months, patients with PBC had significantly lowered levels of anti-RBD-specific IgG compared to HCs, and the inhibition rates of NAb toward wild-type also declined in individuals with PBC. The detected levels of NAb toward Omicron BA.4/5 were below the positive threshold in patients with PBC and HCs. Laboratory parameters did not significantly correlate with any of the three antibodies. The online survey revealed that 24% of patients with PBC received three COVID-19 vaccines, while 63% were unimmunized. Adverse effect rates after the first, second, and third vaccine doses were 6.1%, 10.3%, and 9.5%, respectively. Unvaccinated patients with PBC were more worried about the safety of the vaccine than those who were vaccinated (P = 0.004). As a result, this study fills the immunological assessment gap in patients with PBC who received inactivated COVID-19 vaccines.

4.
Biochem Biophys Res Commun ; 712-713: 149958, 2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38640731

RESUMO

Hepatic stellate cells (HSCs) perform a significant function in liver regeneration (LR) by becoming active. We propose to investigate if activated HSCs enhance glycolysis via PFKFB3, an essential glycolytic regulator, and whether targeting this pathway could be beneficial for LR. The liver and isolated HSCs of mice subjected to 2/3 partial hepatectomy (PHx) exhibited a significant rise in PFKFB3 expression, as indicated by quantitative RT-PCR analyses and Western blotting. Also, the primary HSCs of mice subjected to PHx have a significant elevation of the glycolysis level. Knocking down PFKFB3 significantly diminished the enhancement of glycolysis by PDGF in human LX2 cells. The hepatocyte proliferation in mice treated with PHx was almost completely prevented when the PFKFB3 inhibitor 3PO was administered, emerging that PFKFB3 is essential in LR. Furthermore, there was a decline in mRNA expression of immediate early genes and proinflammatory cytokines. In terms of mechanism, both the p38 MAP kinase and ERK1/2 phosphorylation in LO2 cells and LO2 proliferation were significantly reduced by the conditioned medium (CM) obtained from LX2 cells with either PFKFB3 knockdown or inhibition. Compared to the control group, isolated hepatocytes from 3PO-treated mice showed decreased p38 MAP kinase and ERK1/2 phosphorylation and proliferation. Thus, LR after PHx involves the activation of PFKFB3 in HSCs, which enhances glycolysis and promotes lactate production, thereby facilitating hepatocyte proliferation via the p38/ERK MAPK signaling pathway.

5.
Clin Chim Acta ; : 119681, 2024 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-38643816

RESUMO

OBJECTIVES: Demyelination and immunocyte-infiltrated lesions have been found in neuro-Behçet's disease (NBD) pathology. Lacking satisfying laboratory biomarkers in NBD impedes standard clinical diagnostics. We aim to explore the ancillary indicators for NBD diagnosis unveiling its potential etiology. METHODS: 28 NBD with defined diagnosis, 29 patients with neuropsychiatric lupus erythematosus, 30 central nervous system idiopathic inflammatory demyelination diseases (CNS-IIDD), 30 CNS infections, 30 cerebrovascular diseases, and 30 noninflammatory neurological diseases (NIND) were retrospectively enrolled. Immunoglobulins (Ig) in serum and cerebral spinal fluid (CSF) were detected by immunonephelometry and myelin basic protein (MBP) by quantitative enzyme-linked immunosorbent assay. RESULTS: IgA index is almost twice enhanced in NBD than NIND with an accuracy of 0.8488 in differential diagnosis, the sensitivity and specificity of which were 75.00 % and 90.00 % when the cutoff was > 0.6814. The accuracy of CSF Ig and quotient of Ig all exceed 0.90 in discerning NBD with damaged and intact blood-brain barrier (BBB). Clustering analyses divided NBD into two different phenotypes: one with BBB damage has lower Ig synthesis, the other with extra-synthesis in parenchymal sites but with intact BBB. MBP index is significantly correlated with kappa (KAP) index and lambda (LAM) index (r = 0.358, 0.575, P < 0.001), hinting the NBD pathogenesis of CNS demyelination in triggering excessive intrathecal Ig productions and humoral responses. CONCLUSIONS: IgA index acts as a potential diagnostic indicator in differentiating NBD from NIND and CNS-IIDD. Excessive immunoglobulin production induced by CNS inflammation and demyelination might be latent immunopathogenesis of NBD.

6.
Front Immunol ; 15: 1384270, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38576620

RESUMO

With the proposal of the "biological-psychological-social" model, clinical decision-makers and researchers have paid more attention to the bidirectional interactive effects between psychological factors and diseases. The brain-gut-microbiota axis, as an important pathway for communication between the brain and the gut, plays an important role in the occurrence and development of inflammatory bowel disease. This article reviews the mechanism by which psychological disorders mediate inflammatory bowel disease by affecting the brain-gut-microbiota axis. Research progress on inflammatory bowel disease causing "comorbidities of mind and body" through the microbiota-gut-brain axis is also described. In addition, to meet the needs of individualized treatment, this article describes some nontraditional and easily overlooked treatment strategies that have led to new ideas for "psychosomatic treatment".


Assuntos
Microbioma Gastrointestinal , Doenças Inflamatórias Intestinais , Transtornos Mentais , Microbiota , Humanos , Encéfalo/metabolismo , Doenças Inflamatórias Intestinais/etiologia , Doenças Inflamatórias Intestinais/terapia , Doenças Inflamatórias Intestinais/metabolismo , Transtornos Mentais/metabolismo
7.
Adv Healthc Mater ; : e2304657, 2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38607802

RESUMO

The pervasive employment of antibiotics has engendered the advent of drug-resistant bacteria, imperiling the well-being and health of both humans and animals. Infections precipitated by such multi-resistant bacteria, especially those induced by methicillin-resistant Staphylococcus aureus (MRSA), pervade hospital settings, constituting a grave menace to patient vitality. Antimicrobial peptides (AMPs) have garnered considerable attention as a potent countermeasure against multidrug resistant bacteria. In preceding research endeavors, an insect-derived antimicrobial peptide is identified that, while possessing antimicrobial attributes, manifested suboptimal efficacy against drug-resistant Gram-positive bacteria. To ameliorate this issue, this work enhances the antimicrobial capabilities of the initial ß-hairpin AMPs by substituting the structural sequence of the original AMPs with variant lengths of hydrophobic amino acid-hydrophilic amino acid repeat units. Throughout this endeavor, this work has identified a number of peptides that possess highly effective antibacterial characteristics against a wide range of bacteria. Additionally, some of these peptides have the ability to self-assemble into nanofibers, which then build networks in a distinctive manner to capture bacteria. Consequently, they represent prospective antibiotic alternatives for addressing wound infections engendered by drug-resistant bacteria.

8.
Org Biomol Chem ; 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38651658

RESUMO

An efficient and practical method for the synthesis of 3-alkenylquinoxalinones containing the SCF3 group has been readily developed through a three-component radical cascade reaction involving quinoxalinones, alkynes and AgSCF3. The reaction was found to be compatible with a variety of substrates and exhibited a high functional group tolerance and complete E-selectivity. The preliminary study suggests the involvement of a SCF3 radical in the transformation.

9.
Food Chem ; 449: 139262, 2024 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-38608613

RESUMO

Despite its nutritional components and potential health benefits, the bitterness of quinoa seed limits its utilization in the food industry. Saponins are believed to be the main cause of the bitterness, but it is still uncertain which specific compound is responsible. This study aimed to isolate the main components contributing to the bitterness in quinoa seed by solvent extraction and various column chromatography techniques guided by sensory evaluation. Five compounds were identified by mass spectrometry and nuclear magnetic resonance analyses, with the dose-over-threshold factors from 29.03 to 198.89. The results confirmed that triterpenoids are responsible for the bitter taste in quinoa seed, with phytolaccagenic acid derivatives being the primary contributor. Additionally, kaempferol 3-O-(2″, 6″-di-O-α-rhamnopyranosyl)-ß-galactopyranoside (namely mauritianin), was demonstrated for the first time to be associated with the bitterness of quinoa. This study could provide new insight into the bitter compound identification in quinoa.

10.
Regen Ther ; 27: 244-250, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38586873

RESUMO

Platelet-rich plasma (PRP) has the capability of assisting in the recovery of damaged tissues by releasing a variety of biologically active factors to initiate a hemostatic cascade reaction and promote the synthesis of new connective tissue and revascularization. It is now widely used for tissue engineering repair. In addition, PRP has demonstrated nerve repair and pain relief, and has been studied and applied to the facial nerve, median nerve, sciatic nerve, and central nerve. These suggest that PRP injection therapy has a positive effect on nerve repair. This indicates that PRP has high clinical value and potential application in nerve repair. It is worthwhile for scientists and medical workers to further explore and study PRP to expand its application in nerve repair, and to provide a more reliable scientific basis for the opening of a new approach to nerve repair.

11.
Nanoscale ; 2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38591110

RESUMO

Tumor recurrence after surgical resection remains a significant challenge in breast cancer treatment. Immune checkpoint blockade therapy, as a promising alternative therapy, faces limitations in combating tumor recurrence due to the low immune response rate. In this study, we developed an implantable photo-responsive self-healing hydrogel loaded with MoS2 nanosheets and the immunoadjuvant R837 (PVA-MoS2-R837, PMR hydrogel) for in situ generation of tumor-associated antigens at the post-surgical site of the primary tumor, enabling sustained and effective activation of the immune response. This PMR hydrogel exhibited potential for near-infrared (NIR) light response, tissue adhesion, self-healing, and sustained adjuvant release. When implanted at the site after tumor resection, NIR irradiation triggered a photothermal effect, resulting in the ablation of residual cancer cells. The in situ-generated tumor-associated antigens promoted dendritic cell (DC) maturation. In a mouse model, PMR hydrogel-mediated photothermal therapy combined with immune checkpoint blockade effectively inhibited the recurrence of resected tumors, providing new insights for combating post-resection breast cancer recurrence.

12.
Inflamm Res ; 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38622285

RESUMO

BACKGROUND: Tumor immunotherapy brings new light and vitality to breast cancer patients, but low response rate and limitations of therapeutic targets become major obstacles to its clinical application. Recent studies have shown that CD24 is involved in an important process of tumor immune regulation in breast cancer and is a promising target for immunotherapy. METHODS: In this study, singleR was used to annotate each cell subpopulation after t-distributed stochastic neighbor embedding (t-SNE) methods. Pseudo-time trace analysis and cell communication were analyzed by Monocle2 package and CellChat, respectively. A prognostic model based on CD24-related genes was constructed using several machine learning methods. Multiple quantitative immunofluorescence (MQIF) was used to evaluate the spatial relationship between CD24+PANCK+cells and exhausted CD8+T cells. RESULTS: Based on the scRNA-seq analysis, 1488 CD24-related differential genes were identified, and a risk model consisting of 15 prognostic characteristic genes was constructed by combining the bulk RNA-seq data. Patients were divided into high- and low-risk groups based on the median risk score. Immune landscape analysis showed that the low-risk group showed higher infiltration of immune-promoting cells and stronger immune reactivity. The results of cell communication demonstrated a strong interaction between CD24+epithelial cells and CD8+T cells. Subsequent MQIF demonstrated a strong interaction between CD24+PANCK+ and exhausted CD8+T cells with FOXP3+ in breast cancer. Additionally, CD24+PANCK+ and CD8+FOXP3+T cells were positively associated with lower survival rates. CONCLUSION: This study highlights the importance of CD24+breast cancer cells in clinical prognosis and immunosuppressive microenvironment, which may provide a new direction for improving patient outcomes.

13.
Reprod Sci ; 2024 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-38637474

RESUMO

In women of childbearing age, extensive decidualization, shedding and remodeling of the endometrium during the menstrual cycle are fundamental for successful pregnancy. The role of prostaglandins (PGs) in menstruation has long been proposed in humans, and the rate-limiting enzyme cyclooxygenase was shown to play a key role in endometrial breakdown and shedding in a mouse menstrual-like model in our previous study. However, the specific types of PGs involved and their respective roles remain unclear. Therefore, our objective was to investigate the mechanism through which PGs regulate endometrial disintegration. In this study, the microscopy was observed by HE; the protein levels of prostaglandins E1 (PGE1), prostaglandins E2 (PGE2), prostaglandin F2α (PGF2α) and Prostaglandin I2 (PGI2) were detected by ELISA; the mRNA level of Pfgfr2, Vascular Endothelial Growth Factor(Vegf), Angiostatin and Hypoxia inducible factor-1α (Hif1α) were examined by real-time PCR; PTGFR Receptor (PTGFR), VEGF, Angiostatin and HIF-1α protein levels were investigated by western blotting; the locations of protein were observed by Immunohistochemistry; HIF-1α binding PTGFR promoter was detected by Chromatin Immunoprecipitation (ChIP) and real-time PCR. We found that the concentrations of PGE1, PGE2, and PGF2α all increased significantly during this process. Furthermore, Ptgfr mRNA increased soon after Progesterone (P4) withdrawal, and PTGFR protein levels increased significantly during abundant endometrial breakdown and shedding processes. PTGFR inhibitors AL8810 significantly suppressed endometrial breakdown and shedding, promoted Angiostatin expression, and reduced VEGF-A expressions and vascular permeability. And HIF-1α and PTGFR were mainly located in the luminal/gland epithelium, vascular endothelium, and pre-decidual zone. Interestingly, HIF-1α directly bound to Ptgfr promoter. Moreover, a HIF-1α inhibitor 2-methoxyestradiol (2ME) significantly reduced PTGFR expression and suppressed endometrial breakdown which was in accord with PTGFR inhibitor's effect. Similar changes occurred in human stromal cells relevant to menstruation in vitro. Our study provides evidence that PGF2α/PTGFR plays a vital role in endometrial breakdown via vascular changes that are regulated by HIF-1α during menstruation.

14.
Nat Mater ; 2024 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-38448659

RESUMO

Thrombosis is a leading global cause of death, in part due to the low efficacy of thrombolytic therapy. Here, we describe a method for precise delivery and accurate dosing of tissue plasminogen activator (tPA) using an intelligent DNA nanodevice. We use DNA origami to integrate DNA nanosheets with predesigned tPA binding sites and thrombin-responsive DNA fasteners. The fastener is an interlocking DNA triplex structure that acts as a thrombin recognizer, threshold controller and opening switch. When loaded with tPA and intravenously administrated in vivo, these DNA nanodevices rapidly target the site of thrombosis, track the circulating microemboli and expose the active tPA only when the concentration of thrombin exceeds a threshold. We demonstrate their improved therapeutic efficacy in ischaemic stroke and pulmonary embolism models, supporting the potential of these nanodevices to provide accurate tPA dosing for the treatment of different thromboses.

15.
Heliyon ; 10(5): e27507, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38463870

RESUMO

Background: Malignant pericardial effusion (MPE) is a common complication of advanced breast cancer (BRCA) and plays an important role in BRCA. This study is aims to construct a prognostic model based on MPE-related genes for predicting the prognosis of breast cancer. Methods: The BRCA samples are analyzed based on the expression of MPE-related genes by using an unsupervised cluster analysis method. This study processes the data by least absolute shrinkage and selection operator and multivariate Cox analysis, and uses machine learning algorithms to construct BRCA prognostic model and develop web tool. Results: BRCA patients are classified into three clusters and a BRCA prognostic model is constructed containing 9 MPE-related genes. There are significant differences in signature pathways, immune infiltration, immunotherapy response and drug sensitivity testing between the high and low-risk groups. Of note, a web-based tool (http://wys.helyly.top/cox.html) is developed to predict overall survival as well as drug-therapy response of BRCA patients quickly and conveniently, which can provide a basis for clinicians to formulate individualized treatment plans. Conclusion: The MPE-related prognostic model developed in this study can be used as an effective tool for predicting the prognosis of BRCA and provides new insights for the diagnosis and treatment of BRCA patients.

16.
Small ; : e2311984, 2024 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-38461526

RESUMO

A major issue with Fenton-like reaction is the excessive consumption of H2 O2 caused by the sluggish regeneration rate of low-valent metal, and how to improve the activation efficiency of H2 O2 has become a key in current research. Herein, a nano-heterostructure catalyst (1.0-MnCu/C) based on nano-interface engineering is constructed by supporting Cu and MnO on carbon skeleton, and its kinetic rate for the degradation of tetracycline hydrochloride is 0.0436 min-1 , which is 2.9 times higher than that of Cu/C system (0.0151 min-1 ). The enhancement of removal rate results from the introduced Mn species can aggregate and transfer electrons to Cu sites through the electron bridge Mn-N/O-Cu, thus preventing Cu2+ from oxidizing H2 O2 to form O2 •- , and facilitating the reduction of Cu2+ and generating more reactive oxygen species (1 O2 and ·OH) with stronger oxidation ability, resulting in H2 O2 utilization efficiency is 1.9 times as much as that of Cu/C. Additionally, the good and stable practical application capacity in different bodies demonstrates that it has great potential for practical environmental remediation.

17.
MedComm (2020) ; 5(3): e489, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38469550

RESUMO

Cancer is a major cause of death globally, and traditional treatments often have limited efficacy and adverse effects. Immunotherapy has shown promise in various malignancies but is less effective in tumors with low immunogenicity or immunosuppressive microenvironment, especially sarcomas. Tertiary lymphoid structures (TLSs) have been associated with a favorable response to immunotherapy and improved survival in cancer patients. However, the immunological mechanisms and clinical significance of TLS in malignant tumors are not fully understood. In this review, we elucidate the composition, neogenesis, and immune characteristics of TLS in tumors, as well as the inflammatory response in cancer development. An in-depth discussion of the unique immune characteristics of TLSs in lung cancer, breast cancer, melanoma, and soft tissue sarcomas will be presented. Additionally, the therapeutic implications of TLS, including its role as a marker of therapeutic response and prognosis, and strategies to promote TLS formation and maturation will be explored. Overall, we aim to provide a comprehensive understanding of the role of TLS in the tumor immune microenvironment and suggest potential interventions for cancer treatment.

18.
Mol Biotechnol ; 2024 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-38456963

RESUMO

Precise quantification of human cells in preclinical animal models by a sensitive and specific approach is warranted. The probe-based quantitative PCR (qPCR) assay as a sensitive and swift approach is suitable for the quantification of human cells by targeting human-specific DNA sequences. In this study, we developed an efficient qPCR assay targeting human-specific DNA in ST6GALNAC3 (termed ST6GAL-qPCR) for the quantification of human cells in preclinical animal models. ST6GAL-qPCR probe was synthesized with FAM and non-fluorescent quencher-minor groove binder conjugated to the 5' and 3' end of the probe, respectively. Genomic DNA from human, rhesus monkeys, cynomolgus monkeys, New Zealand White rabbits, SD rats, C57BL/6, and BALB/c mice were utilized for analyzing the specificity and sensitivity of the ST6GAL-qPCR assay. The ST6GAL-qPCR assay targeted human-specific DNA was cloned to pUCM-T vector and released by EcoR I/Hind III digestion for generating a calibration curve. Cell mixing experiment was performed to validate the ST6GAL-qPCR assay by analysis of 0.1%, 0.01%, and 0.001% of human leukocytes mixed with murine thymocytes. The ST6GAL-qPCR assay detected human DNA rather than DNA from the tested animal species. The amplification efficiency of the ST6GAL-qPCR assay was 93% and the linearity of calibration curve was R2 = 0.999. The ST6GAL-qPCR assay detected as low as 5 copies of human-specific DNA and is efficient to specially amplify as low as 30-pg human DNA in the presence of 1 µg of DNA from the tested species, respectively. The ST6GAL-qPCR assay was able to quantify as low as 0.01% of human leukocytes within murine thymocytes. This ST6GAL-qPCR assay can be used as an efficient approach for the quantification of human cells in preclinical animal models.

19.
J Environ Manage ; 356: 120684, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38531133

RESUMO

Microbial biotransformation is a recommended and reliable method in face of formidable tetracycline (TC) with broad-spectrum antibacterial activity. Herein, comprehensive characteristics of a newfound strain and its molecular mechanism in process of TC bioremediation were involved in this study. Specifically, Serratia marcescens MSM2304 isolated from pig manure sludge grew well in presence of TC and achieved optimal removal efficiency of 61% under conditions of initial TC concentration of 10 mg/L, pH of 7.0, cell inoculation amount of 5%, and tryptone of 10 g/L as additional carbon. The pathways of biotransformation include EPS biosorption, cell surface biosorption and biodegradation, which enzymatic processes of biodegradation were occurred through TC adsorbed by biofilms was firstly broken down by extracellular enzymes and part of TC migrated towards biofilm interior and degraded by intracellular enzymes. Wherein extracellular polysaccharides in extracellular polymeric substances (EPS) from biofilm of strain MSM2304 mainly performed extracellular adsorption, and changes in position and intensity of CO, =CH and C-O-C/C-O of EPS possible further implied TC adsorption by it. Biodegradation accounting for 79.07% played a key role in TC biotransformation and could be fitted well by first-order model that manifesting rapid and thorough removal. Potential biodegradation pathway including demethylation, dihydroxylation, oxygenation, and ring opening possibly involved in TC disposal process of MSM2304, TC-degrading metabolites exhibited lower toxicity to indicator bacteria relative to parent TC. Whole genome sequencing as underlying molecular evidence revealed that TC resistance genes, dehydrogenases-encoding genes, monooxygenase-encoding genes, and methyltransferase-encoding genes of strain MSM2304 were positively related to TC biodegradation. Collectively, these results favored a theoretical evaluation for Serratia marcescens MSM2304 as a promising TC-control agent in environmental bioremediation processes.


Assuntos
Serratia marcescens , Tetraciclina , Animais , Suínos , Serratia marcescens/genética , Antibacterianos/análise , Biotransformação , Genômica
20.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 42(1): 67-74, 2024 Feb 01.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-38475953

RESUMO

OBJECTIVES: This clinical study aimed to evaluate the accuracy of a fully digital technique for measuring sagittal condylar inclination (SCI), as well as validating whether differences existed between the left and right SCI values of the same participant, to provide a reference for clinical practice. METHODS: Ten participants with good occlusal relationship and normal temporomandibular joint were recruited. Three methods were used to measure the SCI values of the participants, namely, A (mechanical facebow transferring and mechanical articulator-based measuring method with physical protrusive interocclusal registration), B (face scan-based virtual facebow and virtual articulator-based measuring method with digital protrusive interocclusal registration), and C (jaw motion tracking system-based measuring method). With the group subjected to methods A and C as the control, the SCI values obtained by the three methods were statistically analyzed. The left and right SCI values of the same participant were also compared. RESULTS: The left and right SCI values measured by method A were 41.70°±7.09° and 42.80°±8.62°, those by method B were 35.09°±12.49° and 37.63°±12.10°, and those by method C were 39.43°±8.72° and 38.45°±6.91°. No significant difference existed among the SCI values measured by the three methods (P>0.05). Meanwhile, no statistical difference existed between the SCI values on the left and right sides of the same participant (P>0.05). CONCLUSIONS: The accuracy of the virtual facebow and digital protrusive occlusal registration based SCI measuring method was the same as that of mechanical facebow based and jaw motion tracking system-based methods. The SCI values on the left and right sides of the same participant were similar. Clinically, an appropriate SCI measurement and setting strategy can be selected based on the actual situations.


Assuntos
Côndilo Mandibular , Articulação Temporomandibular , Humanos , Registro da Relação Maxilomandibular/métodos , Articuladores Dentários , Aparelhos de Tração Extrabucal
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