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AIMS: To study the changes in cortical thickness and subcortical gray matter structures in children with complete spinal cord injury (CSCI), reveal the possible causes of dysfunction beyond sensory motor dysfunction after CSCI, and provide a possible neural basis for corresponding functional intervention training. METHODS: Thirty-seven pediatric CSCI patients and 34 age-, gender-matched healthy children as healthy controls (HCs) were recruited. The 3D high-resolution T1-weighted structural images of all subjects were obtained using a 3.0 Tesla MRI system. Statistical differences between pediatric CSCI patients and HCs in cortical thickness and volumes of subcortical gray matter structures were evaluated. Then, correlation analyses were performed to analyze the correlation between the imaging indicators and clinical characteristics. RESULTS: Compared with HCs, pediatric CSCI patients showed decreased cortical thickness in the right precentral gyrus, superior temporal gyrus, and posterior segment of the lateral sulcus, while increased cortical thickness in the right lingual gyrus and inferior occipital gyrus. The volume of the right thalamus in pediatric CSCI patients was significantly smaller than that in HCs. No significant correlation was found between the imaging indicators and the injury duration, sensory scores, and motor scores of pediatric CSCI patients. CONCLUSIONS: These findings demonstrated that the brain structural reorganizations of pediatric CSCI occurred not only in sensory motor areas but also in cognitive and visual related brain regions, which may suggest that the visual processing, cognitive abnormalities, and related early intervention therapy also deserve greater attention beyond sensory motor rehabilitation training in pediatric CSCI patients.
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Córtex Cerebral , Imageamento por Ressonância Magnética , Traumatismos da Medula Espinal , Humanos , Traumatismos da Medula Espinal/patologia , Traumatismos da Medula Espinal/diagnóstico por imagem , Feminino , Masculino , Criança , Adolescente , Córtex Cerebral/diagnóstico por imagem , Córtex Cerebral/patologia , Substância Cinzenta/patologia , Substância Cinzenta/diagnóstico por imagem , Tamanho do ÓrgãoRESUMO
This study aimed to investigate the value of placental real-time shear wave elastography combined with three-dimensional power Doppler index (3D-PDI) in the prediction of preeclampsia. We conducted a retrospective study selecting 60 pregnant women diagnosed with preeclampsia as the experimental group and 60 normal pregnant women as the control group from January 2021 to December 2022. The elastic modulus values of different regions of the placenta and placental 3D-PDI were detected and compared between the two groups. The ROC curve was used to evaluate the diagnostic value of each parameter, alone or in combination, for preeclampsia. The study findings demonstrated that the elastic modulus values of different regions of the placenta and 3D-PDI of the two groups have statistical significance. The values of SWE, VI, FI, and VFI are different in prediction of preeclampsia, and the combination of various parameters can improve the prediction value. Overall, our study provides a valuable method for the prediction of preeclampsia with the advantages of non-invasiveness, efficiency, and simplicity.
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Técnicas de Imagem por Elasticidade , Pré-Eclâmpsia , Gravidez , Feminino , Humanos , Placenta/diagnóstico por imagem , Pré-Eclâmpsia/diagnóstico por imagem , Estudos Retrospectivos , Técnicas de Imagem por Elasticidade/métodos , Ultrassonografia Pré-Natal/métodos , Imageamento Tridimensional/métodos , Ultrassonografia DopplerRESUMO
In this study, we observed pediatric complete spinal cord injury (CSCI) patients receiving MI training and divided them into different groups according to the effect of motor imagery (MI) training on neuropathic pain (NP). Then, we retrospectively analysed the differences in brain structure of these groups before the MI training, identifying brain regions that may predict the effect of MI on NP. Thirty pediatric CSCI patients were included, including 12 patients who experienced NP during MI and 18 patients who did not experience NP during MI according to the MI training follow-up. The 3D high-resolution T1-weighted images of all subjects were obtained using a 3.0 T MRI system before MI training. A two-sample t-test was performed to evaluate the differences in gray matter volume (GMV) between patients who experienced NP and those who did not experience NP during MI. Receiver operating characteristic (ROC) analysis was performed to compute the sensitivity and specificity of the imaging biomarkers for the effect of MI on NP in pediatric CSCI patients. MI evoked NP in some of the pediatric CSCI patients. Compared with patients who did not experience NP, patients who experienced NP during MI showed larger GMV in the right primary sensorimotor cortex (PSMC) and insula. When using the GMV of the right PSMC and insula in combination as a predictor, the area under the curve (AUC) reached 0.824. Our study demonstrated that MI could evoke NP in some pediatric CSCI patients, but not in others. The individual differences in brain reorganization of the right PSMC and insula may contribute to the different effects of MI on NP. Moreover, the GMV of the right PSMC and insula in combination may be an effective indicator for screening pediatric CSCI patients before MI training therapy.
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To enhance the friction and wear properties of alloys, AlCrTiVNbx high-entropy alloys (HEAs) with various Nb contents were prepared using the arc melting technique and then annealed at 1000 °C for 2 h. The microstructure and hardness changes in the AlCrTiVNbx (x = 0.3, 0.4, and 0.5) HEAs after casting and annealing were studied via scanning electron microscopy, X-ray diffractometry, optical microscopy and the Vickers hardness test. The MFT-EC400 ball disc reciprocating friction and wear tester was used to investigate the wear resistance of the HEAs before and after annealing. The results show that the annealed AlCrTiVNbx HEAs changed from a single-phase structure to a multi-phase structure, and the content of the face-center cubic (FCC) phase and hexagonal close-packed (HCP) phase further increases with the increase in Nb content. The hardness value of the annealed HEAs is greatly enhanced compared with the casting state, and the hardness of the Nb0.5 HEA is increased from 543 HV to 725 HV after annealing. The wear resistance of the alloys after the annealing treatment is also greatly improved, among which Nb0.5 has the best wear resistance. The average friction coefficient of Nb0.5 is 0.154 and the wear rate is 2.117 × 10-5 mm3/(N·m). We believe that the precipitation strengthening after the annealing treatment and the lubrication effect of the FCC phase are the reasons for the significant improvement in wear resistance. The morphology of the samples indicates that the wear mechanism of the alloy includes adhesive wear, abrasive wear and a certain degree of oxidation wear.
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BACKGROUND: The alternation of brain white matter (WM) network has been studied in adult spinal cord injury (SCI) patients. However, the WM network alterations in pediatric SCI patients remain unclear. PURPOSE: To evaluate WM network changes and their functional impact in children with thoracolumbar SCI (TSCI). STUDY TYPE: Prospective. SUBJECTS: Thirty-five pediatric patients with TSCI (8.94 ± 1.86 years, 8/27 males/females) and 34 age- and gender-matched healthy controls (HCs) participated in this study. FIELD STRENGTH/SEQUENCE: 3.0 T/DTI imaging using spin-echo echo-planar and T1-weighted imaging using 3D T1-weighted magnetization-prepared rapid gradient-echo sequence. ASSESSMENT: Pediatric SCI patients were evaluated for motor and sensory scores, injury level, time since injury, and age at injury. The WM network was constructed using a continuous tracing method, resulting in a 90 × 90 matrix. The global and regional metrics were obtained to investigate the alterations of the WM structural network. topology. STATISTICAL TESTS: Two-sample independent t-tests, chi-squared test, Mann-Whitney U-test, and Spearman correlation. Statistical significance was set at P < 0.05. RESULTS: Compared with HCs, pediatric TSCI patients displayed decreased shortest path length (Lp = 1.080 ± 0.130) and normalized Lp (λ = 5.020 ± 0.363), and increased global efficiency (Eg = 0.200 ± 0.015). Notably, these patients also demonstrated heightened regional properties in the orbitofrontal cortex, limbic system, default mode network, and several audio-visual-related regions. Moreover, the λ and Lp values negatively correlated with sensory scores. Conversely, nodal efficiency values in the right calcarine fissure and surrounding cortex positively correlated with sensory scores. The age at injury positively correlated with node degree in the left parahippocampal gyrus and nodal efficiency in the right posterior cingulate gyrus. DATA CONCLUSION: Reorganization of the WM networks in pediatric SCI patients is indicated by increased global and nodal efficiency, which may provide promising neuroimaging biomarkers for functional assessment of pediatric SCI. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY: Stage 5.
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Postoperative delirium (POD) is a common and severe complication in elderly patients with hip fractures. Identifying high-risk patients with POD can help improve the outcome of patients with hip fractures. We conducted a retrospective study on elderly patients (≥65 years of age) who underwent orthopedic surgery with hip fracture between January 2014 and August 2019. Conventional logistic regression and five machine-learning algorithms were used to construct prediction models of POD. A nomogram for POD prediction was built with the logistic regression method. The area under the receiver operating characteristic curve (AUC-ROC), accuracy, sensitivity, and precision were calculated to evaluate different models. Feature importance of individuals was interpreted using Shapley Additive Explanations (SHAP). About 797 patients were enrolled in the study, with the incidence of POD at 9.28% (74/797). The age, renal insufficiency, chronic obstructive pulmonary disease (COPD), use of antipsychotics, lactate dehydrogenase (LDH), and C-reactive protein are used to build a nomogram for POD with an AUC of 0.71. The AUCs of five machine-learning models are 0.81 (Random Forest), 0.80 (GBM), 0.68 (AdaBoost), 0.77 (XGBoost), and 0.70 (SVM). The sensitivities of the six models range from 68.8% (logistic regression and SVM) to 91.9% (Random Forest). The precisions of the six machine-learning models range from 18.3% (logistic regression) to 67.8% (SVM). Six prediction models of POD in patients with hip fractures were constructed using logistic regression and five machine-learning algorithms. The application of machine-learning algorithms could provide convenient POD risk stratification to benefit elderly hip fracture patients.
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Delírio do Despertar , Fraturas do Quadril , Idoso , Humanos , Estudos Retrospectivos , Algoritmos , Fraturas do Quadril/cirurgia , Aprendizado de MáquinaRESUMO
OBJECTIVE: To assess the diagnostic value of the W score in differentiating laryngopharyngeal reflux disease (LPRD) patients from the normal population by pharyngeal pH (Dx-pH) monitoring, compared with the RYAN score. METHODS: One hundred and eight patients with suspected LPRD and complete follow-up results after more than 8 weeks of anti-reflux therapy were enrolled from the Department of Otolaryngology-Head and Neck Surgery, Gastroenterology and Respiratory Medicine of seven hospitals. Their Dx-pH monitoring data before treatment were reanalyzed to obtain the W score in addition to the RYAN score and then the diagnostic sensitivity and specificity were compared and evaluated with reference to the result of anti-reflux therapy. RESULTS: In eighty-seven (80.6%) cases, anti-reflux therapy was effective, and in 21 patients (19.4%), therapy was ineffective. Twenty-seven patients (25.0%) had a positive RYAN score. The W score was positive in 79 (73.1%) patients. There were 52 patients who had a negative RYAN score, but a positive W score. The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of the RYAN score were 28.7%, 90.5%, 92.6%, and 23.5%, respectively (kappa = 0.092, P = 0.068), whereas those of the W score for LPRD was 83.9%, 71.4%, 92.4%, and 51.7%, respectively (kappa = 0.484, P < 0.001). CONCLUSIONS: W score is much more sensitive for the diagnosis of LPRD. Prospective studies with larger patient populations are necessary to validate and improve diagnostic efficacy. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR1800014931.
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Lightweight high-entropy alloys (HEAs) are a new class of low-density, high strength-to-weight ratio metallic structural material. Understanding their corrosion behavior is crucial for designing microstructures for their practical applications. This work investigates the electrochemical corrosion behavior of lightweight HEAs AlCrTiV0.5Cux (x = 0, 0.2, 0.4, 0.6, 0.8, and 1.0) in a 0.6 M NaCl solution. These HEAs were produced by vacuum arc melting. In contrast to 304L stainless steel, all of the alloys exhibited lower current density levels caused by self-corrosion, with AlCrTiV0.5 demonstrating the highest corrosion resistance (0.131 µA/cm2). Corrosion resistance decreased along with the content of copper because copper segregation accelerated local corrosion throughout the alloy.
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Objective: To investigate the curative effect of ball tip technology in pedicle screw placement in the patients with degenerative scoliosis (DS), as compared to traditional freehand technique. Methods: A total of 90 patients with degenerative scoliosis who were admitted to Affiliated Hospital of Hebei Engineering University from October 2019 to October 2021 were selected as the objects in this prospective study. They were randomly divided into an experimental group and a control group with 45 cases in each. The clinical indications, the accuracy of pedicle screw placement, the occurrence of surgical complications, the measurement of spinal and pelvic parameters, the recovery of spinal function and pain degree were recorded and compared within the two groups. Results: After treatment, the operation time, intraoperative blood loss, total number of screws, and time of screwing were compared between the two groups, and the difference was not significant (P > .05). However, the bedding time and the hospital stay were shorter in the experimental group than the control group with difference (P < .05). There was no significant difference in clinical standards and poor implantation in the Gertzbein-Robbins A-E classification between the two groups (P > .05). While the number of perfect placement of screws in the experimental group was higher (P < .05). Before treatment, the Cobbs angle and pelvic incidence-lumbar lordosis (PI-LL) levels of the two groups were comparable (P > .05); after treatment, the Cobbs angle and PI-LL levels of the two groups were lower than those before treatment, and the difference was significant (P < .05). There was no significant difference in Cobbs angle and PI-LL levels between groups (P > .05). Before treatment, the JOA and DOI scores of the two groups were comparable (P > .05); after treatment, the JOA and DOI scores of the two groups were improved (P < .05); the improvement of JOA and DOI scores of the experimental group were better than those in the control group (P < .05). Before treatment, there was no significant difference in the pain degree between the two groups (P > .05); after treatment, the pain of the two groups was improved compared with that before treatment, and the pain degree of the experimental group was lower than that of the control group (P < .05). The incidence of postoperative complications in the experimental group was lower than that in the control group, but there was no significant difference in the total incidence of postoperative complications between the two groups (P > .05). Conclusion: The scouting technique-assisted screw placement can effectively improve the spinal function of patients with degenerative scoliosis, with obvious curative effect and high safety.
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Parafusos Pediculares , Escoliose , Humanos , Parafusos Pediculares/efeitos adversos , Complicações Pós-Operatórias , Estudos Prospectivos , Estudos Retrospectivos , Escoliose/cirurgia , Escoliose/complicações , Tecnologia , Resultado do TratamentoRESUMO
This study aims to investigate the brain gray matter volume (GMV) alterations of pediatric complete thoracolumbar spinal cord injury (SCI) without fracture or dislocation (SCIWOFD) using voxel-based morphometry (VBM) analysis and assess the sensitive neuroimaging biomarkers that may be surrogate targets to enhance brain plasticity. A total of 52 pediatric subjects (age range, 6-12 years), including 25 pediatric SCIWOFD patients and 27 typically developing (TD) children were recruited. An independent two-sample t test was performed to assess between-group differences of brain GMV. Partial correlation analyses were performed to explore the correlations between GMV values and The International Standards for Neurological Classification of Spinal Cord Injury scores, age at the time of injury, time after initial SCI. Receiver operating characteristic analysis was performed to compute the sensitivity and specificity of the imaging biomarkers for pediatric SCIWOFD diagnosis. As for the results, pediatric SCIWOFD patients showed significantly decreased GMV of bilateral cerebellum lobule VIII, right middle occipital gyrus and putamen (PUT), left pallidum (PAL) and thalamus, and increased GMV of vermis III, right cerebellum lobule VI, and supramarginal gyrus. In addition, GMV of left PAL and right PUT were negatively correlated with the pinprick/light touch sensory scores in pediatric SCIWOFD patients. Finally, when using the GMV values of left PAL and right PUT in combination as the predictor, area under the curve reached the highest-0.93. These findings provided evidence that the brain undergoes GMV changes after pediatric SCIWOFD, which may suggest important targets for functional remodeling after SCI in children and provide valuable information for the development of novel and effective rehabilitation therapies in the future.
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Fraturas Ósseas , Traumatismos da Medula Espinal , Humanos , Criança , Substância Cinzenta/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Encéfalo/diagnóstico por imagem , Traumatismos da Medula Espinal/diagnóstico por imagem , Lobo ParietalRESUMO
Since late 2019, the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the resultant spread of COVID-19 have given rise to a worldwide health crisis that is posing great challenges to public health and clinical treatment, in addition to serving as a formidable threat to the global economy. To obtain an effective tool to prevent and diagnose viral infections, we attempted to obtain human antibody fragments that can effectively neutralize viral infection and be utilized for rapid virus detection. To this end, several human monoclonal antibodies were isolated by bio-panning a phage-displayed human antibody library, Tomlinson I. The selected clones were demonstrated to bind to the S1 domain of the spike glycoprotein of SARS-CoV-2. Moreover, clone A7 in Fab and IgG formats were found to effectively neutralize the binding of S protein to angiotensin-converting enzyme 2 in the low nM range. In addition, this clone was successfully converted to quench-based fluorescent immunosensors (Quenchbodies) that allowed antigen detection within a few minutes, with the help of a handy fluorometer.
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Bacteriófagos , Técnicas Biossensoriais , COVID-19 , Enzima de Conversão de Angiotensina 2 , Anticorpos Monoclonais , Anticorpos Neutralizantes , Anticorpos Antivirais , Bacteriófagos/metabolismo , COVID-19/diagnóstico , Humanos , Imunoensaio , Fragmentos de Imunoglobulinas , Imunoglobulina G , Glicoproteínas de Membrana/metabolismo , Biblioteca de Peptídeos , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Proteínas do Envelope Viral/metabolismoRESUMO
Objective: This prospective study is aimed at observing the number of nasal itching and sneezing in rats from the macroscopic level and examine the pathological changes of nasal mucosa, Th1 and Th2-related cytokines, and Treg/Th17 by vitamin D3 administration from the microscopic level, in order to explore the role of vitamin D in allergic rhinitis and to provide theoretical guidance for prevention and treatment. Results: There were significant differences in nasal itching and sneezing between the administration groups and the positive groups. Meanwhile, the level of Th1 and Treg in the administration groups increased, while the level of Th2 and Th17 decreased, indicating that the balance of Th1/Th2 was corrected. Our study revealed that vitamin D3 has preventive and therapeutic effects on allergic rhinitis, which provides theoretical guidance for practical application.
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Rinite Alérgica , Linfócitos T Reguladores , Animais , Colecalciferol/farmacologia , Colecalciferol/uso terapêutico , Citocinas/uso terapêutico , Modelos Animais de Doenças , Estudos Prospectivos , Prurido , Ratos , Rinite Alérgica/tratamento farmacológico , Rinite Alérgica/prevenção & controle , Espirro , Vitamina D/farmacologia , Vitamina D/uso terapêuticoRESUMO
Estrogens are effective for stimulating several functions in living organisms and for regulating cancer development by promoting cell proliferation. Estradiol can disrupt the reproductive and endocrine systems, leading to the development of various diseases. In this study, the monoclonal antibody ESC9 was developed by immunizing mice with a 17ß-estradiol (E2) conjugate, preparing an antibody phage display library, and screening monoclonal antibodies from the prepared library. An antibody with the same sequence as that of ESC9 has not been reported previously. The equilibrium dissociation constant between ESC9 and E2 was found to be 43.3 nM. Additionally, we generated an ESC9-derived immunosensor named as the ESC9 Quenchbody (Q-body), which can rapidly and sensitively detect E2. The assay can be completed within 2 min with a limit of detection of 3.9 pg/ml and half-maximal effective concentration of 154.0 ng/ml. Serum E2 levels were measured using the ESC9 Q-body without pretreatment with serum and with a high recovery rate of 83.3-126.7%. The Q-body immunosensor shows potential for clinical applications based on its excellent detection speed and sensitivity.
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OBJECTIVE: Microglial activation is a vital process in the neuroinflammatory response induced by I/R injury. It has been reported that myocyte enhancer factor (MEF)2D expression in activated microglia is associated with microglia-induced inflammatory responses and plays an important role in neuronal survival. This research aimed to investigate the role and mechanism of MEF2D in microglial activation and neuroinflammation in cerebral I/R in vitro and in vivo. METHODS: A cerebral I/R model was established. In vitro, neuronal, or microglial cells were exposed to oxygen-glucose deprivation and reoxygenation to mimic I/R. MEF2D overexpression was induced, and siRNA was administered in vitro and in vivo. Microglial polarization; MEF2D, nuclear transcription factor (NF)-κb, TLR4, and cytokine levels; neuronal injury; mitochondrial function; brain injury and cognitive function were detected in the different groups in vitro and in vivo. RESULTS: We found that oxygen-glucose deprivation increased MEF2D expression in a time-dependent manner in BV2 cells and primary microglia. MEF2D overexpression inhibited microglial activation, the expression of NF-κb and TLR, cytokine levels, and neuronal injury in microglia exposed to oxygen-glucose deprivation and reoxygenation. In the middle cerebral artery occlusion model, microglial activation, the neuroinflammatory response, mitochondrial dysfunction, brain injury, and cognitive function were improved by MEF2D overexpression and aggravated by MEF2D siRNA treatment. CONCLUSION: These results indicate that MEF2D is a necessary molecule for neuroinflammation regulation and neuronal injury in cerebral ischemia.
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Isquemia Encefálica , Fatores de Transcrição MEF2/metabolismo , Microglia/metabolismo , Neuroproteção , Traumatismo por Reperfusão , Animais , Células Cultivadas , Citocinas/metabolismo , Modelos Animais de Doenças , NF-kappa B/metabolismo , RNA Interferente Pequeno , Ratos Sprague-Dawley , Receptor 4 Toll-Like/metabolismoRESUMO
BACKGROUND: Kabuki syndrome (KS) is a rare syndrome characterized by multisystem congenital anomalies and developmental disorder. KMT2D and KDM6A mutations were identified as the main causative genes in KS patients. There are few case reports and genetic analyses, especially of KDM6A gene mutation, in China. CASE SUMMARY: This study reports a de novo KDM6A mutation in a Chinese infant with KS. A 2-month-old Chinese baby was diagnosed with KS, which manifested as hypoglycemia, congenital anal atresia at birth, feeding difficulties, hypotonia, and serious postnatal growth retardation. He died of recurrent respiratory infections at age 13 mo. DNA sequencing of his blood DNA revealed a novel KDM6A frameshift mutation (c.704_705delAG, p. N236Sfs*26) (GRCh37/hg19). CONCLUSION: We present a Chinese KS patient with a novel KDM6A frameshift mutation (c.704_705delAG, p. N236Sfs*26) (GRCh37/hg19), broadening the mutation spectrum.
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Tumor necrosis factor α (TNF-α) is used as a biomarker for the diagnosis of various inflammatory and autoimmune diseases. In recent years, numerous approaches have been used for the qualitative and quantitative analyses of TNF-α. However, these methods have several drawbacks, such as a tedious and time-consuming process, high pH and temperature sensitivity, and increased chances of denaturation in vitro. Quenchbody (Q-body) is a fluorescence immunoprobe that functions based on the principle of photoinduced electron transfer and has been successful in detecting various substances. In this study, we constructed two Q-bodies based on a therapeutic antibody, adalimumab, to rapidly detect human TNF-α. Both sensors could detect TNF-α within 5 min. The results showed that the limit of detection (LOD) of TNF-α was as low as 0.123 ng/mL with a half-maximal effective concentration (EC50) of 25.0 ng/mL using the TAMRA-labeled Q-body, whereas the ATTO520-labeled Q-body had a LOD of 0.419 ng/mL with an EC50 of 65.6 ng/mL, suggesting that the Q-bodies could rapidly detect TNF-α with reasonable sensitivity over a wide detection range. These biosensors will be useful tools for the detection and monitoring of inflammatory biomarkers.
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BACKGROUND: The detection and identification of single nucleotide polymorphism (SNP) is essential for determining patient disease susceptibility and the delivery of medicines targeted to the individual. At present, SNP genotyping technology includes Sanger sequencing, TaqMan-probe quantitative polymerase chain reaction (qPCR), amplification-refractory mutation system (ARMS)-PCR, and Kompetitive Allele-Specific PCR (KASP). However, these technologies have some disadvantages: the high cost of development and detection, long and time consuming protocols, and high false positive rates. Focusing on these limitations, we proposed a new SNP detection method named universal probe-based intermediate primer-triggered qPCR (UPIP-qPCR). In this method, only two types of fluorescence-labeled probes were used for SNP genotyping, thus greatly reducing the cost of development and detection for SNP genotyping. RESULTS: In the amplification process of UPIP-qPCR, unlabeled intermediate primers with template-specific recognition functions could trigger probe hydrolysis and specific signal release. UPIP-qPCR can be used successfully and widely for SNP genotyping. The sensitivity of UPIP-qPCR in SNP genotyping was 0.01 ng, the call rate was more than 99.1%, and the accuracy was more than 99.9%. High-throughput DNA microarrays based on intermediate primers can be used for SNP genotyping. CONCLUSION: This novel approach is both cost effective and highly accurate; it is a reliable SNP genotyping method that would serve the needs of the clinician in the provision of targeted medicine.
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Técnicas de Genotipagem , Polimorfismo de Nucleotídeo Único , Alelos , Genótipo , Humanos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
We successfully developed a fluorescent probe that can quickly convert full-length antibodies to Quenchbodies, which represent a type of fluorescent immunosensor with high binding affinity and specificity depending on the reaction of antigens and antibodies. An anti-testosterone IgG was successfully converted to an immunosensor that detects testosterone with a limit of detection (LOD) of 0.76 nM and concentration for 50% of maximal effect (EC50) of 61.5 nM. Another IgG-based immunosensor detected ractopamine with an LOD of 15.5 pM and EC50 of 48.6 nM.
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Técnicas Biossensoriais , Proteínas de Transporte , Imunoensaio , Imunoglobulina G , Limite de DetecçãoRESUMO
BACKGROUND: Although many studies have suggested that air cells may play an important role in sigmoid sinus diverticulum and/or dehiscence (SSDD)-induced pulsatile tinnitus (PT), the exact effects remain unclear. This study aims to quantitatively investigate the effect of different degrees of pneumatization of temporal bone on sound transmission from the sigmoid sinus to the tympanic cavity. METHODS: In the clinical study, 25 patients were enrolled to measure the sound intensity and frequency of SSDD-induced PT. In the experimental study, sound inputs at different frequencies at the dehiscent sigmoid plate were measured and compared among realistic object models of different degrees of pneumatization, when sound intensity in the tympanic cavity was fixed as the level of the PT sound intensity sensed by patients. RESULTS: The sound intensity sensed by PT patients was 34.0 ± 13.0 dB SPL, which represented the sound intensity in the tympanic cavity transmitted from the dehiscent sigmoid plate. In the experimental study, when sound received in the tympanic cavity was fixed at 34 dB SPL, the mean inputs of sound intensity at the dehiscent sigmoid plate were 46.9, 46.2, 45.2, 47.1, 57, 57.4, and 74.1 dB SPL in a hypo-pneumatized model; 42.6, 43, 41.5, 43.2, 47.3, 58.2, and 78.8 dB SPL in a moderately pneumatized model; 52.6, 52.8, 48.1, 61, 64.2, 82.4, and >87.3 dB SPL in a well-pneumatized model; and 47.2, 46.2, 45.4, 49.4, 54.9, 66.6, and 77.7 dB SPL in hyper-pneumatized model, with increased sound frequencies (125-8000 Hz). The mean sound transmission distances were 41.7 mm, 45.2 mm, 47.6 mm, and 48.4 mm in successively better pneumatized models. CONCLUSION: Sound reduces while passing through air cells and attenuation is lowest in the moderately pneumatized temporal bone, followed by the hypo- and hyper-pneumatized temporal bones, and the highest in the well-pneumatized temporal bone. Lower-frequency sound attenuation (≤1000 Hz) is less than that of higher-frequency sound (>1000 Hz).
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Divertículo , Osso Temporal , Zumbido , Cavidades Cranianas/diagnóstico por imagem , Humanos , Som , Osso Temporal/diagnóstico por imagem , Zumbido/etiologiaRESUMO
Antibody phage display technology plays an important role in the development of monoclonal antibodies, humanization, and affinity evolution of antibodies. Thus far, antibody phage display mainly focuses on the display of antibody variable region or antigen-binding fragments. In this study, we constructed a new phage display system that can display full-length IgG antibodies on M13 phage. The phage display vector contains open reading frames (ORFs) encoding full-length the heavy and light chains of the antibody. NcoI/XhoI restriction enzyme sites were used to clone the variable region of the heavy chain into the heavy chain ORF, and SalI/NotI sites were used to clone the light chain variable region. SnaBI and SbfI restriction enzyme sites were designed between the cloning sites of heavy and light chains, respectively, to increase the cloning efficiency. The full-length antibodies of nivolumab against programmed death factor 1, trastuzumab against human epidermal growth factor 2, diL2K against the cluster of differentiation 3 epsilon, and adalimumab against tumor necrosis factor- alpha were displayed on phage with the vector. Phage-displayed antibodies showed their original antigen-binding activity. An amber codon shifted the vector to express IgG in non-suppressed Escherichia coli. The heavy and light chains of the E. coli-expressed antibodies could be detected through western blotting, and the antigen-binding activity was confirmed using an enzyme-linked immunosorbent assay. Biopanning was carried out with a model phage display antibody library, and the results showed that the novel phage system could be used for antibody library construction and highly efficient antibody screening. The reported system is the first full-length antibody phage display system.