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BACKGROUND: The successful treatment and improvement of acute kidney injury (AKI) depend on early-stage diagnosis. However, no study has differentiated between the three stages of AKI and non-AKI patients following heart surgery. This study will fill this gap in the literature and help to improve kidney disease management in the future. METHODS: In this study, we applied Raman spectroscopy (RS) to uncover unique urine biomarkers distinguishing heart surgery patients with and without AKI. Given the amplified risk of renal complications post-cardiac surgery, this approach is of paramount importance. Further, we employed the partial least squares-support vector machine (PLS-SVM) model to distinguish between all three stages of AKI and non-AKI patients. RESULTS: We noted significant metabolic disparities among the groups. Each AKI stage presented a distinct metabolic profile: stage 1 had elevated uric acid and reduced creatinine levels; stage 2 demonstrated increased tryptophan and nitrogenous compounds with diminished uric acid; stage 3 displayed the highest neopterin and the lowest creatinine levels. We utilized the PLS-SVM model for discriminant analysis, achieving over 90% identification rate in distinguishing AKI patients, encompassing all stages, from non-AKI subjects. CONCLUSIONS: This study characterizes the incidence and risk factors for AKI after cardiac surgery. The unique spectral information garnered from this study can also pave the way for developing an in vivo RS method to detect and monitor AKI effectively.
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Atteinte rénale aigüe , Marqueurs biologiques , Procédures de chirurgie cardiaque , Analyse spectrale Raman , Examen des urines , Humains , Atteinte rénale aigüe/diagnostic , Atteinte rénale aigüe/urine , Atteinte rénale aigüe/étiologie , Analyse spectrale Raman/méthodes , Procédures de chirurgie cardiaque/effets indésirables , Mâle , Femelle , Adulte d'âge moyen , Sujet âgé , Marqueurs biologiques/urine , Examen des urines/méthodes , Créatinine/urine , Machine à vecteur de support , Acide urique/urine , Complications postopératoires/diagnostic , Complications postopératoires/urine , Complications postopératoires/étiologie , Facteurs de risque , Méthode des moindres carrésRÉSUMÉ
Accurate identification of tissue types in surgical margins is essential for ensuring the complete removal of cancerous cells and minimizing the risk of recurrence. The objective of this study was to explore the clinical utility of Raman spectroscopy for the detection of oral squamous cell carcinoma (OSCC) in both tumor and healthy tissues obtained from surgical resection specimens during surgery. This study enrolled a total of 64 patients diagnosed with OSCC. Among the participants, approximately 50% of the cases were classified as the most advanced stage, referred to as T4. Raman experiments were conducted on cryopreserved tissue samples collected from patients diagnosed with OSCC. Prominent spectral regions containing key oral biomarkers were analyzed using the partial least squares-support vector machine (PLS-SVM) method, which is a powerful multivariate analysis technique for discriminant analysis. This approach effectively differentiated OSCC tissue from non-OSCC tissue, achieving a sensitivity of 95.7% and a specificity of 93.3% with 94.7% accuracy. In the current study, Raman analysis of fresh tissue samples showed that OSCC tissues contained significantly higher levels of nucleic acids, proteins, and several amino acids compared to the adjacent healthy tissues. In addition to differentiating between OSCC and non-OSCC tissues, we have also explored the potential of Raman spectroscopy in classifying different stages of OSCC. Specifically, we have investigated the classification of T1, T2, T3, and T4 stages based on their Raman spectra. These findings emphasize the importance of considering both stage and subsite factors in the application of Raman spectroscopy for OSCC analysis. Future work will focus on expanding our tissue sample collection to better comprehend how different subsites influence the Raman spectra of OSCC at various stages, aiming to improve diagnostic accuracy and aid in identifying tumor-free margins during surgical interventions.
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Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic worldwide. Currently, however, no effective drug or vaccine is available to treat or prevent the resulting coronavirus disease 2019 (COVID-19). Here, we report our discovery of a promising anti-COVID-19 drug candidate, the lipoglycopeptide antibiotic dalbavancin, based on virtual screening of the FDA-approved peptide drug library combined with in vitro and in vivo functional antiviral assays. Our results showed that dalbavancin directly binds to human angiotensin-converting enzyme 2 (ACE2) with high affinity, thereby blocking its interaction with the SARS-CoV-2 spike protein. Furthermore, dalbavancin effectively prevents SARS-CoV-2 replication in Vero E6 cells with an EC50 of ~12 nM. In both mouse and rhesus macaque models, viral replication and histopathological injuries caused by SARS-CoV-2 infection are significantly inhibited by dalbavancin administration. Given its high safety and long plasma half-life (8-10 days) shown in previous clinical trials, our data indicate that dalbavancin is a promising anti-COVID-19 drug candidate.
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Angiotensin-converting enzyme 2/métabolisme , Antiviraux , SARS-CoV-2/métabolisme , Glycoprotéine de spicule des coronavirus/métabolisme , Téicoplanine/analogues et dérivés , Animaux , Antiviraux/pharmacocinétique , Antiviraux/pharmacologie , Cellules Caco-2 , Chlorocebus aethiops , Modèles animaux de maladie humaine , Humains , Souris , Souris transgéniques , Liaison aux protéines/effets des médicaments et des substances chimiques , Téicoplanine/pharmacocinétique , Téicoplanine/pharmacologie , Cellules VeroRÉSUMÉ
A surface acoustic wave (SAW) sensor was investigated for its application in C-reactive protein (CRP) detection. Piezoelectric lithium niobate (LiNbO3) substrates were used to study their frequency response characteristics in a SAW sensor with a CRP sensing area. After the fabrication of the SAW sensor, the immobilization process was performed for CRP/anti-CRP interaction. The CRP/anti-CRP interaction can be detected as mass variations in the sensing area. These mass variations may produce changes in the amplitude of sensor response. It was clearly observed that a CRP concentration of 0.1 µg/mL can be detected in the proposed SAW sensor. A good fitting linear relationship between the detected insertion loss (amplitude) and the concentrations of CRP from 0.1 µg/mL to 1 mg/mL was obtained. The detected shifts in the amplitude of insertion loss in SAW sensors for different CRP concentrations may be useful in the diagnosis of risk of cardiovascular diseases.
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Protéine C-réactive , Maladies cardiovasculaires , Son (physique) , Protéine C-réactive/analyse , Maladies cardiovasculaires/diagnosticRÉSUMÉ
BACKGROUND: Oral cancer is one of the most common diseases globally. Conventional oral examination and histopathological examination are the two main clinical methods for diagnosing oral cancer early. VELscope is an oral cancer-screening device that exploited autofluorescence. It yields inconsistent results when used to differentiate between normal, premalignant and malignant lesions. We develop a new method to increase the accuracy of differentiation. MATERIALS AND METHODS: Five samples (images) of each of 21 normal mucosae, as well as 31 premalignant and 16 malignant lesions of the tongue and buccal mucosa were collected under both white light and autofluorescence (VELscope, 400-460 nm wavelength). The images were developed using an iPod (Apple, Atlanta Georgia, USA). RESULTS: The normalized intensity and standard deviation of intensity were calculated to classify image pixels from the region of interest (ROI). Linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA) classifiers were used. The performance of both of the classifiers was evaluated with respect to accuracy, precision, and recall. These parameters were used for multiclass classification. The accuracy rate of LDA with un-normalized data was increased by 2% and 14% and that of QDA was increased by 16% and 25% for the tongue and buccal mucosa, respectively. CONCLUSION: The QDA algorithm outperforms the LDA classifier in the analysis of autofluorescence images with respect to all of the standard evaluation parameters.
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Traitement d'image par ordinateur , Tumeurs de la bouche/imagerie diagnostique , Imagerie optique , Adulte , Analyse discriminante , Femelle , Humains , Mâle , Muqueuse de la bouche/imagerie diagnostiqueRÉSUMÉ
The light emitting diode (LED) is widely used in modern solid-state lighting applications, and its output efficiency is closely related to the submounts' material properties. Most submounts used today, such as low-power printed circuit boards (PCBs) or high-power metal core printed circuit boards (MCPCBs), are not transparent and seriously decrease the output light extraction. To meet the requirements of high light output and better color mixing, a three-dimensional (3-D) stacked flip-chip (FC) LED module is proposed and demonstrated. To realize light penetration and mixing, the mentioned 3-D vertically stacking RGB LEDs use transparent glass as FC package submounts called glass circuit boards (GCB). Light emitted from each GCB stacked LEDs passes through each other and thus exhibits good output efficiency and homogeneous light-mixing characteristics. In this work, the parasitic problem of heat accumulation, which caused by the poor thermal conductivity of GCB and leads to a serious decrease in output efficiency, is solved by a proposed transparent cooling oil encapsulation (OCP) method.
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Monolithic phosphor-free two-color gallium nitride (GaN)-based white light emitting diodes (LED) have the potential to replace current phosphor-based GaN white LEDs due to their low cost and long life cycle. Unfortunately, the growth of high indium content indium gallium nitride (InGaN)/GaN quantum dot and reported LED's color rendering index (CRI) are still problematic. Here, we use flip-chip technology to fabricate an upside down monolithic two-color phosphor-free LED with four grown layers of high indium quantum dots on top of the three grown layers of lower indium quantum wells separated by a GaN tunneling barrier layer. The photoluminescence (PL) and electroluminescence (EL) spectra of this white LED reveal a broad spectrum ranging from 475 to 675 nm which is close to an ideal white-light source. The corresponding color temperature and color rendering index (CRI) of the fabricated white LED, operated at 350, 500, and 750 mA, are comparable to that of the conventional phosphor-based LEDs. Insights of the epitaxial structure and the transport mechanism were revealed through the TEM and temperature dependent PL and EL measurements. Our results show true potential in the Epi-ready GaN white LEDs for future solid state lighting applications.
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Consistent NDVI time series are basic and prerequisite in long-term monitoring of land surface properties. Advanced very high resolution radiometer (AVHRR) measurements provide the longest records of continuous global satellite measurements sensitive to live green vegetation, and moderate resolution imaging spectroradiometer (MODIS) is more recent typical with high spatial and temporal resolution. Understanding the relationship between the AVHRR-derived NDVI and MODIS NDVI is critical to continued long-term monitoring of ecological resources. NDVI time series acquired by the global inventory modeling and mapping studies (GIMMS) and Terra MODIS were compared over the same time periods from 2000 to 2006 at four scales of Qinghai-Tibet Plateau (whole region, sub-region, biome and pixel) to assess the level of agreement in terms of absolute values and dynamic change by independently assessing the performance of GIMMS and MODIS NDVI and using 495 Landsat samples of 20 km x20 km covering major land cover type. High correlations existed between the two datasets at the four scales, indicating their mostly equal capability of capturing seasonal and monthly phenological variations (mostly at 0. 001 significance level). Simi- larities of the two datasets differed significantly among different vegetation types. The relative low correlation coefficients and large difference of NDVI value between the two datasets were found among dense vegetation types including broadleaf forest and needleleaf forest, yet the correlations were strong and the deviations were small in more homogeneous vegetation types, such as meadow, steppe and crop. 82% of study area was characterized by strong consistency between GIMMS and MODIS NDVI at pixel scale. In the Landsat NDVI vs. GIMMS and MODIS NDVI comparison of absolute values, the MODIS NDVI performed slightly better than GIMMS NDVI, whereas in the comparison of temporal change values, the GIMMS data set performed best. Similar with comparison results of GIMMS and MODIS NDVI, the consistency across the three datasets was clearly different among various vegetation types. In dynamic changes, differences between Landsat and MODIS NDVI were smaller than Landsat NDVI vs. GIMMS NDVI for forest, but Landsat and GIMMS NDVI agreed better for grass and crop. The results suggested that spatial patterns and dynamic trends of GIMMS NDVI were found to be in overall acceptable agreement with MODIS NDVI. It might be feasible to successfully integrate historical GIMMS and more recent MODIS NDVI to provide continuity of NDVI products. The accuracy of merging AVHRR historical data recorded with more modern MODIS NDVI data strongly depends on vegetation type, season and phenological period, and spatial scale. The integration of the two datasets for needleleaf forest, broadleaf forest, and for all vegetation types in the phenological transition periods in spring and autumn should be treated with caution.
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Surveillance de l'environnement , Forêts , Plantes , Chine , Écologie , Imagerie satellitaire , Saisons , Analyse spatio-temporelle , Analyse spectraleRÉSUMÉ
OBJECTIVE: To investigate sperm DNA integrity in male infertility patients with hepatitis B virus (HBV) infection. METHODS: This study included 90 infertile men with HBV infection (group A), 82 infertile men without HBV infection (group B) and 70 normal fertile men (group C). We detected sperm DNA integrity among the subjects, including DNA fragmentation index (DFI) and high DNA stainability (HDS), by sperm chromatin structure assay (SCSA), and compared them among the three groups. RESULTS: DFI was higher in group A ([28.17 +/- 13.06]%) than in B ([26.64 +/- 9.79]%) and C ([15.67 +/- 4.73]%), significantly higher in A and B than in C (P < 0.05) but with no significant difference between A and B (P > 0.05). HDS was higher in group A ([10.83 +/- 5.601]%) than in B ([9.04 +/- 3.48]%) and C ([8.04-2.25]%), with significant difference between A and C (P < 0.05). CONCLUSION: Sperm DNA integrity of infertile males is significantly different from that of normal fertile men, and infertility with HBV infection further impairs sperm DNA, which is manifested by abnormal sperm nuclear maturity.
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ADN/génétique , Hépatite B/anatomopathologie , Infertilité masculine/génétique , Infertilité masculine/virologie , Adulte , Études cas-témoins , Chromatine , Altération de l'ADN , Virus de l'hépatite B , Humains , Mâle , Numération des spermatozoïdes , Spermatozoïdes/anatomopathologie , Jeune adulteRÉSUMÉ
Detection of unlabeled oligonucleotides using surface plasmon resonance (SPR) is difficult because of the oligonucleotides' relatively lower molecular weight compared with proteins. In this paper, we describe a method for detecting unlabeled oligonucleotides at low concentration using a paired surface plasma waves biosensor (PSPWB). The biosensor uses a sensor chip with an immobilized probe to detect a target oligonucleotide via sequence-specific hybridization. PSPWB measures the demodulated amplitude of the heterodyne signal in real time. In the meantime, the ratio of the amplitudes between the detected output signal and reference can reduce the excess noise from the laser intensity fluctuation. Also, the common-path propagation of p and s waves cancels the common phase noise induced by temperature variation. Thus, a high signal-to-noise ratio (SNR) of the heterodyne signal is detected. The sequence specificity of oligonucleotide hybridization ensures that the platform is precisely discriminating between target and non-target oligonucleotides. Under optimized experimental conditions, the detected heterodyne signal increases linearly with the logarithm of the concentration of target oligonucleotide over the range 0.5-500 pM. The detection limit is 0.5 pM in this experiment. In addition, the non-target oligonucleotide at concentrations of 10 pM and 10nM generated signals only slightly higher than background, indicating the high selectivity and specificity of this method. Different length of perfectly matched oligonucleotide targets at 10-mer, 15-mer and 20-mer were identified at the concentration of 150 pM.
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Oligonucléotides/analyse , Résonance plasmonique de surface/méthodes , Séquence nucléotidique , Limite de détection , Hybridation d'acides nucléiques , Sondes oligonucléotidiques/génétique , Oligonucléotides/génétique , Sensibilité et spécificité , Rapport signal-bruit , Résonance plasmonique de surface/statistiques et données numériquesRÉSUMÉ
In this study, we applied the developed paired surface plasma waves biosensor (PSPWB) in a dual-channel biosensor for rapid and sensitive detection of swine-origin influenza A (H1N1) virus (S-OIV). In conjunction with the amplitude ratio of the signal and the reference channel, the stability of the PSPWB system is significantly improved experimentally. The theoretical limit of detection (LOD) of the dual-channel PSPWB for S-OIV is 30 PFU/mL (PFU, plaque-forming unit), which was calculated from the fitting curve of the surface plasmon resonance signal with a S-OIV clinical isolate concentration in phosphate-buffered saline (PBS) over a range of 18-1.8 × 10(6) PFU/mL. The LOD is 2 orders of magnitude more sensitive than the commercial rapid influenza diagnostic test at worst and an order of magnitude less sensitive than real-time quantitative polymerase chain reaction (PCR) whose LOD for S-OIV in PBS was determined to be 3.5 PFU/mL in this experiment. Furthermore, under in vivo conditions, this experiment demonstrates that the assay successfully measured S-OIV at a concentration of 1.8 × 10(2) PFU/mL in mimic solution, which contained PBS-diluted normal human nasal mucosa. Most importantly, the assay time took less than 20 min. From the results, the dual-channel PSPWB potentially offers great opportunity in developing an alternative PCR-free diagnostic method for rapid, sensitive, and accurate detection of viral pathogens with epidemiological relevance in clinical samples by using an appropriate pathogen-specific antibody.
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Sous-type H1N1 du virus de la grippe A/isolement et purification , Infections à Orthomyxoviridae/diagnostic , Infections à Orthomyxoviridae/médecine vétérinaire , Résonance plasmonique de surface/instrumentation , Suidae/virologie , Animaux , Conception d'appareillage , Humains , Limite de détection , Résonance plasmonique de surface/économie , Résonance plasmonique de surface/méthodes , Facteurs tempsRÉSUMÉ
Measuring the kinetic constants of protein-protein interactions at ultralow concentrations becomes critical in characterizing biospecific affinity, and exploring the feasibility of clinical diagnosis with respect to detection sensitivity, efficiency and accuracy. In this study, we propose a method that can calculate the binding constants of protein-protein interactions in sandwich assays at ultralow concentrations at the pg/mL level, using a localized surface plasmon coupled fluorescence fiber-optic biosensor (LSPCF-FOB). We discuss a two-compartment model to achieve reaction-limited kinetics under the stagnant conditions of the reaction chamber. The association rate constant, dissociation rate constant, and the equilibrium dissociation constant, that is, k(a), k(d), K(D), respectively, of the kinetics of binding between total prostate-specific antigen (t-PSA) and anti-t-PSA at concentrations from 0.1 pg/mL to 1 ng/mL, were measured either in PBS or in human serum. This is the first time that k(a), k(d), and K(D) have been measured at such a low concentration range in a complex sample such as human serum.
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Techniques de biocapteur/méthodes , Or/composition chimique , Nanoparticules métalliques/composition chimique , Animaux , Anticorps immobilisés/immunologie , Techniques de biocapteur/instrumentation , Bovins , Humains , Immunoglobuline G/immunologie , Cinétique , Limite de détection , Souris , Fibres optiques , Liaison aux protéines , Spectrométrie de fluorescence , Spécificité du substratRÉSUMÉ
Swine-origin influenza A (H1N1) virus (S-OIV) was identified as a new reassortant strain of influenza A virus in April 2009 and led to an influenza pandemic. Accurate and timely diagnoses are crucial for the control of influenza disease. We developed a localized surface plasmon coupled fluorescence fiber-optic biosensor (LSPCF-FOB) which combines a sandwich immunoassay with the LSP technique using antibodies against the hemagglutinin (HA) proteins of S-OIVs. The detection limit of the LSPCF-FOB for recombinant S-OIV H1 protein detection was estimated at 13.9 pg/mL, which is 10(3)-fold better than that of conventional capture ELISA when using the same capture antibodies. For clinical S-OIV isolates measurement, meanwhile, the detection limit of the LSPCF-FOB platform was calculated to be 8.25 × 10(4)copies/mL, compared with 2.06 × 10(6)copies/mL using conventional capture ELISA. Furthermore, in comparison with the influenza A/B rapid test, the detection limit of the LSPCF-FOB for S-OIV was almost 50-fold in PBS solution and 25-fold lower in mimic solution, which used nasal mucosa from healthy donors as the diluent. The findings of this study therefore indicate that the high detection sensitivity and specificity of the LSPCF-FOB make it a potentially effective diagnostic tool for clinical S-OIV infection and this technique has the potential to be applied to the development of other clinical microbe detection platforms.
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Sous-type H1N1 du virus de la grippe A/isolement et purification , Fibres optiques , Virus recombinants/isolement et purification , Résonance plasmonique de surface/instrumentation , Suidae/virologie , Animaux , Anticorps antiviraux , Séquence nucléotidique , Amorces ADN/génétique , Test ELISA , Fluorescence , Glycoprotéine hémagglutinine du virus influenza/analyse , Glycoprotéine hémagglutinine du virus influenza/génétique , Glycoprotéine hémagglutinine du virus influenza/immunologie , Humains , Sous-type H1N1 du virus de la grippe A/génétique , Sous-type H1N1 du virus de la grippe A/immunologie , Grippe humaine/diagnostic , Grippe humaine/épidémiologie , Grippe humaine/virologie , Pandémies , Virus recombinants/génétique , Protéines recombinantes/analyse , Protéines recombinantes/génétique , Protéines recombinantes/immunologie , RT-PCR , Sensibilité et spécificité , Résonance plasmonique de surface/méthodes , Résonance plasmonique de surface/statistiques et données numériquesRÉSUMÉ
In this study, we demonstrated that an amplitude-sensitive paired surface plasma wave biosensor (PSPWB) is capable of real-time detection of prostate-specific antigen (PSA) in diluted human serum without labeling. Experimentally, the detection limit of PSPWB was 8.4 x 10(-9) refractive index unit (RIU) and the PSPWB could measure PSA in a phosphate buffered saline solution from 10 fg/mL ( approximately 300 aM) to 100 pg/mL ( approximately 3 pM) successfully, with demonstration of a linear relationship between PSA concentrations and surface plasmon resonance (SPR) signals. Therefore, results were obtained over a wide dynamic range 5 orders of magnitude for analyte concentration. In addition, the PSPWB successfully detected PSA in diluted human serum as well. These experimental results indicate that the PSPWB is capable of detection with high sensitivity over a wide range by using SPR-based biosensors and has a capability of detecting biological analytes in clinical sample without complicated operating procedures.
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Techniques de biocapteur/méthodes , Antigène spécifique de la prostate/sang , Résonance plasmonique de surface , Humains , Limite de détection , Mâle , Résonance plasmonique de surface/méthodesRÉSUMÉ
This research proposed a dual-frequency heterodyne ellipsometer (DHE) in which a dual-frequency collinearly polarized laser beam with equal amplitude and zero phase difference between p- and s-polarizations is setup. It is based on the polarizer-sample-analyzer, PSA configuration of the conventional ellipsometer. DHE enables to characterize a generalized elliptical phase retarder by treating it as the combination of a linear phase retarder and a polarization rotator. The method for measuring elliptical birefringence of an elliptical phase retarder based on the equivalence theorem of an unitary optical system was derived and the experimental verification by use of DHE was demonstrated too. The experimental results show the capability of DHE on characterization of a generalized phase retardation plate accurately.
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In this study, we demonstrated that the fiber-optic biosensor based on localized surface plasmon coupled fluorescence (LSPCF) is capable of detecting alpha-fetoprotein (AFP) in human serum. The sensitivity of LSPCF fiber-optic biosensor is not only enhanced but also the specific selectivity is improved since the fluorophores are excited by the localized surface plasmon with high efficiency. Experimentally, this fiber-optic biosensor is able to detect AFP concentration in phosphate buffered saline (PBS) solution from 0.1ng/mL to 100ng/mL whereas the linear relationship between the AFP concentrations and the fluorescence signals is shown. Furthermore, a linear response between the fluorescence signals and the concentrations of AFP in human serum from 2.33ng/mL to 143.74ng/mL is also obtained. As a result, the detection limit of the LSPCF fiber-optic biosensor on AFP detection is comparable with the conventional enzyme-linked immunosorbent assay (ELISA). Additionally, the LSPCF fiber-optic biosensor benefits on inexpensive, disposable and simpler optical geometry that can become a high efficient immunoassay comparable with the conventional ELISA and radioimmunoassay (RIA) clinically.
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Techniques de biocapteur/instrumentation , Analyse chimique du sang/instrumentation , Technologie des fibres optiques/instrumentation , Dosage immunologique/instrumentation , Spectrométrie de fluorescence/instrumentation , Résonance plasmonique de surface/instrumentation , Alphafoetoprotéines/analyse , Conception d'appareillage , Analyse de panne d'appareillage , Humains , Reproductibilité des résultats , Sensibilité et spécificitéRÉSUMÉ
In this paper, a novel differential-phase-sensitive surface plasmon resonance biosensor (DP-SPRB) is proposed and developed, in which a two-frequency laser is integrated with a differential amplifier in order to analytically convert the phase modulation into amplitude modulation. With the use of the conventional envelope detection technique, the differential phase is precisely decoded in real time in terms of the demodulated amplitude. In order to verify high detection sensitivity of the DP-SPRB, a sucrose-water solution and glycerin-water solution at low concentrations were both tested, and the experimental results confirm that the detection sensitivity on wt % concentration of the sucrose solution is 0.00001%. Moreover, the real-time monitoring mouse IgG/antimouse IgG interaction shows the minimum concentration of mouse IgG to be at 10 fg/mL. To our knowledge, this is the highest sensitivity ever measured by a surface plasmon resonance biosensor. However, because of the limited dynamic range of DP-SPRB, it can only apply to biomolecule interactions at extremely low concentration.
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Techniques de biocapteur/instrumentation , Techniques de biocapteur/méthodes , Résonance plasmonique de surface/instrumentation , Résonance plasmonique de surface/méthodes , Glycérol , Immunoglobuline G/analyse , Immunoglobuline G/immunologie , Solutions , Saccharose , EauRÉSUMÉ
Objective To construct recombinant eukaryotic expression plasmid encoding Swedish and Flemish mutations of amyloid precursor protein (APP) fused with fluorescent protein and to investigate the APP cleavage progress. Methods The last 300 bases of APP (named as C99 containing Flemish mutation), together with cyan and yellow fluorescence sequence (named as CFP and YFP,respectively) were obtained by polymerase chain reaction (PCR). The 54 bases in the middle of APP sequence were synthesized (named as 54 bp containing Swedish mutation). The 4 fragments mentioned above (CFP, YFP, C99 as well as 54 bp) were inserted into the vector pcDNA3.0. By genetic engineering, the recombinant plasmid pcDNA3.0-CFP-54bp-YFP-C99 was constructed and identified by enzyme digestion, PCR and sequencing. Then the plasmid was transfected into SH-SY5Y cells. Its expression was examined by fluorescence confocal microscopy and the fluorescence resonance energy transfer (FRET) signal was collected. The amyloid beta (A) deposition was detected by immunocytochemistry. Results (1) DNA sequencing showed the sequence of the constructed recombinant plasmid was correct. (2) FRET and two types of fluorescence could be seen by the spectrum confocal fluorescence microscopy. (3) The expression product of fusion gene was correct and cleaved by and secretases. (4) The A deposition was detected in the cell membrane, cytoplasma and intercellular space. Conclusion (1) The fusion protein can generate A by and γproteolytic processing. (2) It is for the first time to observe the APP cleavage by FRET. (3) It is also the first time to find that APP may be cleaved during its transportation from cell plasma to cell membrane. (4) C99 is very important for the correct cleavage of APP. Our test data strongly suggest that C99 may function as the signal peptide. It might guide and direct the APP to the right location for the cleavage.
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Objective To observe the effect of movement exercise combined with electroaeupuneture on the expression of Nestin in the hippocampus dentate gyrus (DG) after cerebral ischemia-repeffusion.Methods Fifty- four Wistar rats were used and randomly divided into a control group (Group A),an exercise training group (Group B),and an exercise training combined with electroacupuncture group (Group C).The middle cerebral arteries (MCA) of all the rats were occluded for 1 h,followed by reperfusion for 7,14 and 21 davs.Immunohistochemistry method was used to detect the expression of Nestin in the hippoeampus dentate gyrus.Results The number of Nes- tin-positive cells peaked in DG in all groups on the 7th day after cerebral isehemia-reperfusion.The number of Nes- tin-positive cells in DG ipsilateral to the ischemia-reperfusion lesion were significantly more than those in the opposite side at various time points (P