RESUMO
In recent years, optical tweezers have become an effective bioassay tool due to their unique advantages, especially in combination with suspension beads, which can be applied to develop a high-performance analysis platform capable of high-quality imaging and stable signal output. However, the optical tweezer-assisted bead analysis is still at the early stage, and further development of different favorable methods is in need. Herein, we have first developed the optical tweezer-assisted immuno-rolling circle amplification (immuno-RCA) on beads for protein detection. Prostate-specific antigen was selected as the model analyte, and the immunosandwich structure on beads was built by the high affinity of "antibody-antigen". The "protein-nucleic acid" signals were effectively converted through the covalent coupling procedure of antibodies and oligonucleotides, further initiating the RCA reaction to achieve signal amplification. The individual beads with the strong irregular Brownian motion in a fluid environment were eventually trapped by the optical tweezers to acquire the accurate and high-quality signal. Compared with the conventional immunoassay on beads, the sensitivity of the developed strategy was increased by 587 times with a limit of detection of 4.29 pg/mL (0.13 pM), as well as excellent specificity, stability, and reproducibility. This study developed the new optical tweezer-assisted beads imaging strategy for protein targets, which has great potential for being applied to clinical serology research and expands the application of optical tweezers in the bioassays.
Assuntos
Pinças Ópticas , Antígeno Prostático Específico , Antígeno Prostático Específico/análise , Humanos , Técnicas de Amplificação de Ácido Nucleico , Imunoensaio/métodos , Limite de Detecção , Microesferas , Técnicas Biossensoriais/métodosRESUMO
Controlling the spread of pathogen requires an efficient and accurate diagnosis. Compared with nucleic acid and antibody detection, antigen assays are more convenient to meet clinical diagnostic needs. However, antigen detection is often difficult to achieve high sensitivity in a limited time. In this work, a novel aptasensing method was designed for the purpose of SARS-CoV-2 antigen detection, using a dumbbell padlock probe-mediated circle-to-circle amplification (C2CA) approach. A sandwich complex of antibody-antigen-aptamer is first formed on the magnetic beads. Afterwards, the signal is amplified by a C2CA reaction involving two tandem rolling circle amplifications. Without special instruments or nanomaterials, a detection limit of 575 fg/mL for S1 protein can be achieved in less than 2 h. In the case of the spike pseudovirus SARS-CoV-2 in artificial saliva, the detection limit is 272 TU/µL, which is much lower than average viral load in patients. Therefore, our method provides a timely, efficient and accurate approach for the clinical diagnosis of SARS-CoV-2. It also opens up the application of C2CA in aptamer sensing and antigen detection.
Assuntos
Técnicas Biossensoriais , COVID-19 , Técnicas Biossensoriais/métodos , COVID-19/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , SARS-CoV-2RESUMO
In this study, phase change materials (PCMs) were innovatively incorporated into hybrid fiber concrete. The properties of PCMs, which absorb and release heat during phase transitions, enable the concrete to actively respond to complex and varying temperature environments. This integration reduces the internal temperature differentials within the concrete, thereby preventing temperature-induced cracks in deep wellbore structures. Through the temperature control model test of the frozen shaft wall, it can be seen that the hybrid fiber phase change concrete (HFPCC) significantly reduces the internal temperature difference, and the maximum temperature difference along the radial direction is 35.84% lower than that of benchmark concrete (BC). The numerical simulation results indicate that a moderate phase transition temperature should be selected in engineering. The phase change temperature should not be close to the ambient temperature and peak temperature. The peak temperature can be reduced by 9.32% and the maximum radial temperature difference can be reduced by 30.89% by selecting an appropriate phase change temperature. The peak temperature and radial maximum temperature difference are both proportional to the latent heat of phase change. The temperature control performance of phase change concrete can be further improved by increasing the latent heat of phase change materials.
Assuntos
Materiais de Construção , Transição de Fase , Congelamento , Modelos Teóricos , Teste de Materiais , TemperaturaRESUMO
OBJECTIVE: To develop and validate a predictive model for sarcopenia. METHODS: A total of 240 subjects who visited our hospital between August 2021 and May 2023 were randomly divided by time of entry into a training set containing 2/3 of patients and a validation set containing 1/3 of patients. The muscle thickness (MT), echo intensity (EI), and shear wave velocity (SWV) of the medial gastrocnemius muscle were measured. Indicators that were meaningful in the univariate analysis in the training set were included in a binary logistic regression to derive a regression model, and the model was evaluated using a consistency index, calibration plot, and clinical validity curve. Diagnostic efficacy and clinical applicability were compared between the model and unifactorial indicators. RESULTS: Four meaningful variables, age, body mass index (BMI), MT, and SWV, were screened into the predictive model. The model was Logit Y = 21.292 + 0.065 × Age - 0.411 × BMI - 0.524 × MT - 3.072 × SWV. The model was well differentiated with an internally validated C-index of 0.924 and an external validation C-index of 0.914. The calibration plot predicted probabilities against actual probabilities showed excellent agreement. The specificity, sensitivity, and Youden's index of the model were 73.80%, 97.40%, and 71.20%, respectively, when using the diagnostic cut-off value of >0.279 for sarcopenia. The logistic model had higher diagnostic efficacy (p < 0.001) and higher net clinical benefit (p < 0.001) over the same threshold range compared to indicators. CONCLUSION: The logistic model of sarcopenia has been justified to have good discriminatory, calibrated, and clinical validity, and has higher diagnostic value than indicators.
Assuntos
Técnicas de Imagem por Elasticidade , Músculo Esquelético , Sarcopenia , Humanos , Sarcopenia/diagnóstico por imagem , Masculino , Feminino , Músculo Esquelético/diagnóstico por imagem , Pessoa de Meia-Idade , Idoso , Técnicas de Imagem por Elasticidade/métodos , Ultrassonografia/métodos , Reprodutibilidade dos Testes , Valor Preditivo dos Testes , AdultoRESUMO
Background: Leukemia patients undergoing cryopreserved ovarian tissue transplantation (OTT) may carry a high risk of disease induction. Measurable residual disease (MRD) in bone marrow is linked to an elevated risk of relapse. It is controversial whether leukemia patients must be allowed to achieve measurable residual disease negative (MRD-negative) status instead of measurable residual disease positive (MRD-positive) status before ovarian tissue cryopreservation (OTC). Objective: To explore the safety and efficacy of OTT in acute leukemia patients with different MRD status by using xenotransplantation. Method: Cryopreserved ovarian tissue from 19 leukemia patients was thawed and xenotransplanted to ovariectomized BALB/C nude mice (n=36). The mice were divided into 2 groups based on the patient's MRD status before OTC: MRD-negative group (n=18) and MRD-positive group (n=18), additionally, a control group consisted of ovariectomized mice (n=9). Body weight was measured weekly and mortality, emaciation, and other abnormalities were recorded. Twenty-six weeks post-surgery, livers, spleens, uteruses, and ovarian grafts were removed for macroscopic and histological examinations to evaluate the efficacy of xenotransplantation and assess malignant cell contamination in mice. Results: Follicle growth was visible in the ovarian grafts of the MRD-negative and MRD-positive groups. Compared with the ovariectomized group, a significant decrease in body weight (p<0.01) was noted, the uterine volume was notably larger, estradiol (E2) levels were significantly higher (p<0.01), and follicle-stimulating hormone (FSH) levels were significantly lower (p<0.001) in the other two groups. Mice in the MRD-positive group showed a significantly higher incidence of death (p<0.001) and emaciation (p<0.01), compared to the MRD-negative group. Histological observation revealed the presence of malignant cells in the grafts, livers, and spleens of 3 mice in the MRD-positive group. No abnormalities were observed in the mice from the MRD-negative group in both macroscopic and histological observations except one mouse was sacrificed for ascites unrelated to leukemia relapse. Conclusion: For leukemia patients having ovarian tissue preserved in the first and only centralized human ovarian tissue cryobank in China, immunodeficient mice xenotransplantation can be a method to evaluate the safety and efficacy of OTT; the risk of malignant cell reimplantation due to OTT is higher in leukemia patients with MRD-positive status than those with MRD-negative status before OTC.