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OBJECTIVES: The previously established 38-plex InDel system was optimized and its performance was validated according to the Scientific Working Group on DNA Analysis Method (SWGDAM) application guidelines. The ancestry inference accuracy of individuals from East Asian, European, African and mixed populations was verified. METHODS: DNA standard sample 9947A was used as the template to establish the optimal amplification conditions by adjusting primer balance, Mg2+ final concentration and optimizing PCR thermal cycle parameters and amplification volume. The allelic dropout, nonspecific amplification and whether the origin of the inferred samples matched the known information were compared to evaluate the performance of this system. RESULTS: The optimal dosage of this system was 0.125-2 ng DNA template. The results of InDel typing were accurate, the amplification equilibrium was good, and the species specificity was good. This system showed certain tolerance to DNA samples including the inhibitor such as hemoglobin (≤80 µmol/L), indigo (≤40 mmol/L), calcium ion (≤1.0 mmol/L), and humic acid (≤90 ng/µL). The system enabled the direct amplification of DNA from saliva and blood on filter paper, and the results of ethnic inference were accurate. The system successfully detected the mixed DNA sample from two individuals. The test results of the system for common biological materials in practical cases were accurate. CONCLUSIONS: The results of the 38-plex InDel system are accurate and reliable, and the performance of the system meets the requirement of the SWGDAM guidelines. This system can accurately differentiate the ancestry origins of individuals from African, European, East Asian, and Eurasian populations and can be implemented in forensic practice.
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DNA , Polimorfismo de Nucleotídeo Único , Humanos , Genótipo , Reação em Cadeia da Polimerase , DNA/genética , Impressões Digitais de DNA/métodos , Mutação INDEL , Genética Populacional , Frequência do GeneRESUMO
Receptor like protein kinases (RLKs) play vital roles in both plant development and stress conditions. Using drought-treated "Yugu 1" as materials, a drought-responsive RLK gene, SiRLK35, was isolated through iTRAQ analysis. In this study, the further analyses of the gene functions were carried out. First, full-length SiRLK35 was amplified by PCR using the cDNA of foxtail millet seedlings as a template. The expression patterns of SiRLK35 under NaCl, PEG, ABA, GA and MeJA treatment were analyzed by quantitative real-time PCR (qRT-PCR), and we found that the expression of SiRLK35 could be induced under different treatments, especially under NaCl treatment. Second, the prokaryotic expression plasmid of SiRLK35 was constructed, and the salt resistance of SiRLK35 was detected by the bacterial plaque growth method. And we uncovered that the growth and tolerance of SiRLK35-containing Escherichia coli strains were in better conditions than control under the NaCl stress. Lastly, pCambia1301P-SiRLK35 was constructed and transformed into rice to obtain transgenic plants. The tolerance of transgenic rice plants to salt stress was higher than that of controls through physiological analysis. We propose that SiRLK35 may participate in salt and stress resistance processes, which could provide potential theoretical foundation for the stress resistance varieties cultivation and breeding of foxtail millet.
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Proteínas de Plantas/genética , Setaria (Planta)/genética , Clonagem Molecular/métodos , Regulação da Expressão Gênica de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plântula/genética , Cloreto de Sódio/metabolismo , Estresse Fisiológico/genéticaRESUMO
Immunogenic antigen (jujuboside A-BSA) and coating antigen (jujuboside A-OVA) of jujuboside A were synthesized by sodium periodate oxidation method for the first time. Jujuboside A artificial antigen was confirmed by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). The titer and specificity of the antibody in serum of immunized mice were detected by enzyme-linked immunosorbent assay (ELISA). The corrected relation curve of inhibition rate showed that the antibody against Jujuboside A obtained from immunized mice could bind to jujuboside A and the titer was up to 1â¶4 000. The jujuboside A artificial antigen was synthesized, which can be used further to preparation of monoclonal antibody and the pharmacokinetics study of jujuboside A in laboratory animals.
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Antígenos/química , Saponinas/síntese química , Animais , Ensaio de Imunoadsorção Enzimática , Camundongos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
A laser-diode-pumped dual-loss-modulated Q-switching Tm:LuAG laser with an acousto-optic modulator (AOM) and monolayer graphene saturable absorber (SA) around 2 µm is presented for the first time to the best of our knowledge. The average output power and the pulse widths for different repetition rates have been measured. In comparison with the singly Q-switching laser with AOM or with monolayer graphene SA, the dual-loss-modulated Q-switching laser could generate shorter pulse width and higher peak power. The maximum pulse width compression ratio was found to be 3.11, and the highest peak power was enhanced 97.4 times. The experimental results show that the dual-loss-modulated technology is an efficient method for compressing the pulse width, improving the peak power, and enhancing the pulse stability for the Q-switched lasers at 2 µm.
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Grafite/química , Lasers de Estado Sólido , Acústica , Alumínio , Desenho de Equipamento , Lasers , Lutécio , Dispositivos Ópticos , Oscilometria , TúlioRESUMO
OBJECTIVE: To establish an enzyme linked immunosorbent assay(ELISA) for mangiferin (MRn), artificial antigen of MRn was synthesized. METHODS: Oxidation method using sodium iodide was used to synthesize immunogenic antigen (MRn-BSA) and coating antigen(MRn-OVA) of MRn. The characterization of the synthesis was examined by UV spectrometry. BALB/c mice were immunized with the prepared MRn-BSA immunogenic antigen. The titer of the anti-serum was detected by ELISA, in the meanwhile the immunogenicity of MRn-BSA was confirmed by indirect competitive ELISA (icELISA). RESULTS: UV spectroscopy showed that MRn was successfully conjugated with OVA and BSA,so the new compound of MRn-BSA was synthesized, the coupling ratio was 6: 1. After immuned MRn-BSA, the mice could produce anti-MRn antibodies specifically, of which titer was up to 1: 4 000, and the general range was 1 - 100 µg/mL. CONCLUSION: Anti-MRn antibodies are discovered in the serum of mice, which indicates that artificial antigen of MRn is successfully synthesized,for the purpose of preparing monoclonal antibodies, as well as the establishment of appropriate immune methods.
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Antígenos/química , Ensaio de Imunoadsorção Enzimática , Xantonas/imunologia , Animais , Anticorpos/sangue , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine (TCM) and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF-MS) and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1:8000. The artificial antigen of catalpol was successfully synthesized.
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Antígenos/imunologia , Glucosídeos Iridoides/imunologia , Animais , Anticorpos/imunologia , Antígenos/química , Imunoensaio , Glucosídeos Iridoides/química , Masculino , Medicina Tradicional Chinesa , Camundongos , Camundongos Endogâmicos BALB C , Soroalbumina Bovina/química , Soroalbumina Bovina/imunologiaRESUMO
Geniposide, Geniposide, the main active component in extracts of Gardenia jasminoides ELLIS., is one of the main components of Huanglian-Jiedu-Tang (HJT). This study aimed to validate an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on monoclonal antibodies (mAb) against geniposide (anti-geniposide mAb), which was developed by our lab, and apply the assay to study the pharmacokinetics of geniposide in HJT in mice. Blood samples were drawn from mice at predetermined time points after oral administration of HJT in three dosages. A linear correlation was obtained for geniposide concentrations in the range from 1.17 to 37.50 µg/mL. The intra-day and inter-day precision values of the icELISA method were well within the recommended range (≤10%). The recovery rates ranged from 99.74 to 102.40%. Stability studies showed that geniposide sample solutions were intact for 12 h. The Tmax and mean residence time (MRT) of geniposide of the three groups were consistent with previous data. The results suggest that a reliable and effective method was established and could be applied to the study of the pharmacokinetics of geniposide in HJT.
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Medicamentos de Ervas Chinesas/farmacocinética , Iridoides/sangue , Administração Oral , Animais , Anticorpos Monoclonais/imunologia , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/química , Ensaio de Imunoadsorção Enzimática , Iridoides/análise , Iridoides/imunologia , Masculino , Camundongos Endogâmicos BALB C , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To observe the preventive effect of two fractionations for postoperative radiotherapy of keloid and discuss the optimal way for postoperative radiotherapy. METHODS: We enrolled 107 consecutive keloid patients with 139 lesions from August 2011 to October 2012 in Department of Radiation Oncology of Peking University First Hospital. There were 114 lesions (the largest lesion part will be accounted if there are several lesions in the single body area) into the curative effect of the statistics. All the patients received irradiation after operation within 24 hours. The patients were divided into two groups: 5 Gy/f for continuous 4 days (5 Gy group); 4 Gy/f for continuous 5 days (4 Gy group). The lesions were treated by 6 MeV-E by Varian 21EX medical linear accelerator made in America. The irradiation field was surgical incision plus 1 cm in radial directions. One centimeter bolus was put on the skin to attain the therapeutical dose of skin surface. The total dose for each lesion was 20 Gy. The treatment effect of keloid was classified into cure, excellence and recurrence, referring to Darzi's standard. Effectivity means the sum of cure and excellence. SPSS 14.0 was used to statistically analyze the data. RESULTS: The total effective rate for 5 Gy group was 90.7% (49/54) and 66.7% (40/60) for 4 Gy group (P = 0.001). The lesions were divided into three regions according to the tension of the skin: ear/face/neck region, chest wall/shoulder/back region and other regions. The treatment effects of 5 Gy group and 4 Gy group were 94.1% (16/17) vs. 85.0% (17/20) for ear/face/neck region, 89.7% (26/29) vs. 60.0% (18/30) for chest wall/shoulder/back region and 87.5% (7/8) vs. 50.0% (5/10) for other regions. Significant difference was found in chest wall/shoulder/back region (P = 0.009). No obvious toxicities occurred in any group. CONCLUSION: Postoperative radiation therapy within 24 hours of 5 Gy/f for continuous 4 days and 4 Gy/f for continuous 5 days is effective, especially in 5 Gy/f group. It is suggested that hypofractionated radiation therapy is more effective for keloid patients, and it is also economical and convenient for patients and worth further discussing.
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Fracionamento da Dose de Radiação , Queloide/radioterapia , Humanos , Período Pós-Operatório , RecidivaRESUMO
Oxidation method with sodium iodide was used to synthesize immunogenic antigen (PF-BSA) and coating antigen (PF-OVA) of paeoniflorin. UV spectroscopy showed that paeoniflorin was successfully conjugated with BSA and OVA. After immunized by PF-BSA, the mice can produce anti-paeoniflorin antibodies specifically. The ELISA test results showed the high titer (1:12 800) and specificity (IC50 = 0.791 mg x L(-1)) of the antiserum from mice injected with PF-BSA. Also, the antiserum showed low cross activities against nine traditional Chinese medicine (TCM) of small molecules. These artificial antigens were successfully synthesized and the anti-paeoniflorin antibody well prepared, which provides the experimental basis for the further study of ELISA and its kit.
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Antígenos/química , Medicamentos de Ervas Chinesas/química , Glucosídeos/química , Monoterpenos/química , Animais , Anticorpos/análise , Antígenos/imunologia , Ensaio de Imunoadsorção Enzimática , Glucosídeos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Monoterpenos/imunologia , Soroalbumina Bovina/química , Soroalbumina Bovina/imunologiaRESUMO
The establishment of high specificity and sensitivity method of small molecule monoclonal antibody-based immunoassay has a great importance in the study of small molecule compounds in Chinese medicine, wherein synthesis of small molecule artificial antigen is a critical step in the preparation of small molecule antibodies. Oxidation method using sodium iodide was used to synthesize immunogenic antigen (FRn-BSA) and coating antigen (FRn-OVA) of forsythin. UV spectroscopy and thin layer chromatography showed that forsythin was successfully conjugated with BSA and OVA. After immuned FRn-BSA, the mice could specifically produce anti-forsythin antibodies with titer up to 1:8 000, and the linear range was from 1 mg x L(-1) to 100 mg x L(-1). In this paper, the artificial antigen of forsythin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.
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Antígenos/imunologia , Compostos Bicíclicos Heterocíclicos com Pontes/imunologia , Medicamentos de Ervas Chinesas/química , Furanos/imunologia , Animais , Anticorpos/imunologia , Antígenos/química , Compostos Bicíclicos Heterocíclicos com Pontes/química , Furanos/química , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
This paper proposes an extended state observer (ESO) based data-driven set-point learning control (DDSPLC) scheme for a class of nonlinear batch processes with a priori P-type feedback control structure subject to nonrepetitive uncertainties, by only using the process input and output data available in practice. Firstly, the unknown process dynamics is equivalently transformed into an iterative dynamic linearization data model (IDLDM) with a residual term. A radial basis function neural network is adopted to estimate the pseudo partial derivative information related to IDLDM, and meanwhile, a data-driven iterative ESO is constructed to estimate the unknown residual term along the batch direction. Then, an adaptive set-point learning control law is designed to merely regulate the set-point command of the closed-loop control structure for realizing batch optimization. Robust convergence of the output tracking error along the batch direction is rigorously analyzed by using the contraction mapping approach and mathematical induction. Finally, two illustrative examples from the literature are used to validate the effectiveness and advantage of the proposed design.
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Study on pharmacodynamic material basis of traditional Chinese medicines is one of the key issues for the modernization of traditional Chinese medicine. Having introduced the monoclonal antibody technology into the study on pharmacodynamic material basis of traditional Chinese medicines, the author prepared the immunoaffinity chromatography column by using monoclonal antibodies in active components of traditional Chinese medicines, so as to selectively knock out the component from herbs or traditional Chinese medicine compounds, while preserving all of the other components and keeping their amount and ratio unchanged. A comparative study on pharmacokinetics and pharmacodynamics was made to explicitly reveal the correlation between the component and the main purpose of traditional Chinese medicines and compounds. The analysis on pharmacodynamic material basis of traditional Chinese medicines by using specific knockout technology with monoclonal antibodies is a new method for study pharmacodynamic material basis in line with the characteristics of traditional Chinese medicines. Its results can not only help study material basis from a new perspective, but also help find the modern scientific significance in single herb or among compounds of traditional Chinese medicines.
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Anticorpos Monoclonais/química , Cromatografia de Afinidade/métodos , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Adsorção , Animais , Linhagem Celular , Cromatografia de Afinidade/instrumentação , Medicamentos de Ervas Chinesas/química , HumanosRESUMO
This article presents an event-triggered adaptive NN command-filtered control for a class of multi-input and multi-output (MIMO) nonlinear systems with unknown rate-dependent hysteresis in the actuator and the constraints on full states. The ETM is used to reduce the communication frequency between controller and actuator. The command filter technique is first employed to solve the dilemma between the nondifferentiable control signal at triggering instants and rate-dependent hysteresis input premise while avoiding the "explosion of complexity" problem. During the backstepping design, the barrier Lyapunov functions are utilized to guarantee that system states will stay in certain regions and the unknown nonlinear items are approximated by adaptive neural networks. The compensating signals are constructed to eliminate filtering errors. The estimates of unknown hysteresis parameters are updated by adaptive laws. The stability analysis is given and the effectiveness of the proposed method is verified by simulation.
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This article is concerned with passivity analysis of neural networks with a time-varying delay. Several techniques in the domain are improved to establish the new passivity criterion with less conservatism. First, a Lyapunov-Krasovskii functional (LKF) is constructed with two general delay-product-type terms which contain any chosen degree of polynomials in time-varying delay. Second, a general convexity lemma without conservatism is developed to address the positive-definiteness of the LKF and the negative-definiteness of its time-derivative. Then, with these improved results, a hierarchical passivity criterion of less conservatism is obtained for neural networks with a time-varying delay, whose size and conservatism vary with the maximal degree of the time-varying delay polynomial in the LKF. It is shown that the conservatism of the passivity criterion does not always reduce as the degree of the time-varying delay polynomial increases. Finally, a numerical example is given to illustrate the proposed criterion and benchmark against the existing results.
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This note shows an elegant relationship between the quadratic optimal control and robust stabilization for linear time-invariant (LTI) systems, where the former control can robustly stabilize the latter system, provided that the matched uncertainty is bounded. Through reviewing the relevant literature, some common mistakes in regard to this relationship are found. The correct results are obtained and proved in both frequency and time domains. The results are applicable to both single- and multi-input cases. They are significant as the simple LQR design for the nominal system can be utilized to directly solve-with no further effort-the complex robust stabilization problem for a class of linear uncertain systems.
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Vascular and neurological damage are the typical outcomes of ischemic strokes. Vascular endothelial cells (VECs), a substantial component of the blood-brain barrier (BBB), are necessary for normal cerebrovascular physiology. During an ischemic stroke (IS), changes in the brain endothelium can lead to a BBB rupture, inflammation, and vasogenic brain edema, and VECs are essential for neurotrophic effects and angiogenesis. Non-coding RNAs (nc-RNAs) are endogenous molecules, and brain ischemia quickly changes the expression patterns of several non-coding RNA types, such as microRNA (miRNA/miR), long non-coding RNA (lncRNA), and circular RNA (circRNA). Furthermore, vascular endothelium-associated nc-RNAs are important mediators in the maintenance of healthy cerebrovascular function. In order to better understand how VECs are regulated epigenetically during an IS, in this review, we attempted to assemble the molecular functions of nc-RNAs that are linked with VECs during an IS.
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AVC Isquêmico , MicroRNAs , Acidente Vascular Cerebral , Humanos , Células Endoteliais/metabolismo , Acidente Vascular Cerebral/metabolismo , RNA não Traduzido/genética , RNA não Traduzido/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Endotélio Vascular/metabolismo , RNA Circular/metabolismo , AVC Isquêmico/genéticaRESUMO
Compatibility mechanism study of Chinese herbal compound (CHC) has been one of key contents in Chinese medical research, but the present research methods are not suitable for its own features due to its complexity, which has restricted the process of modernization and intemationalization of Chinese medicine. In this paper, we addressed that the compatibility is closely correlated to their in vivo metabolic processes. The preparation of active small molecules monoclonal antibodies in herbs and testing a variety of effective compositions simultaneously using immunoassay can clarify the in vivo metabolism and mutual interactions of Chinese herbs, which is a new thought of studying the compound compatibility mechanisms.
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Anticorpos Monoclonais/administração & dosagem , Medicamentos de Ervas Chinesas/administração & dosagem , Combinação de Medicamentos , Incompatibilidade de MedicamentosRESUMO
OBJECTIVE: To define the weighting coefficients of the symptoms and signs in the diagnosis of corresponding traditional Chinese medicine (TCM) syndrome elements of ulcerative colitis based on expert questionnaire investigation. METHODS: The questionnaire included eight syndrome elements of location and 11 syndrome elements relating to disease characteristics. Experts who answered the questionnaire were titled as associate chief physician or above, and had been practicing as professionals in general practice of medicine of digestive system for a long time. They came from 15 third-grade class-A hospitals (6 provinces and Beijing) in China. The weighting coefficients of the symptoms and signs in the diagnosis of corresponding TCM syndrome elements of ulcerative colitis were identified preliminarily based on consent degree of the experts (mean value and variation coefficient). RESULTS: There were 9, 6, 14, 5, 9, 8, 12 and 2 symptoms and signs corresponding to 8 TCM syndrome elements related to disease location and the ranges of weighting coefficients were listed as follows: liver, from 7.49 to 4.18; heart, from 6.90 to 5.51; spleen, from 7.96 to 5.55; lung, from 6.30 to 5.27; kidney, from 7.82 to 5.71; stomach, from 7.53 to 6.15; large intestine, from 8.40 to 6.70; bowel collaterals, from 8.49 to 6.41. Numbers of symptoms and signs corresponding to 11 TCM syndrome elements related to disease nature were 13, 8, 17, 18, 17, 18, 6, 11, 2, 10 and 11 and the ranges of weighting coefficients were listed as follows: qi deficiency, from 7.44 to 5.60; blood deficiency, from 7.90 to 5.59; yin deficiency, from 6.88 to 4.91; yang deficiency, from 7.54 to 5.57; dampness, from 7.91 to 4.96; excess heat (fire), from 7.13 to 5.69; excess cold, from 7.51 to 6.14; qi stagnation, from 7.38 to 5.88; qi sinking, from 7.65 to 7.43; blood stasis, from 7.60 to 5.79; fluid retention, from 7.08 to 4.46. CONCLUSION: On the basis of previous document research and clinical research in ulcerative colitis, the expert questionnaire can collect consensus opinions of experts. The weighting coefficients of the symptoms and signs in the diagnosis of corresponding TCM syndrome elements of ulcerative colitis are defined preliminarily. Thereby it will lay the foundation for establishing differentiation criteria of ulcerative colitis based on TCM syndrome elements.
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Colite Ulcerativa/diagnóstico , Medicina Tradicional Chinesa , Médicos , Inquéritos e Questionários , Adulto , Idoso , Diagnóstico Diferencial , Humanos , Pessoa de Meia-Idade , Deficiência da Energia Yang , Deficiência da Energia YinRESUMO
This article investigates the problem of command-filtered event-triggered adaptive fuzzy neural network (FNN) output feedback control for stochastic nonlinear systems (SNSs) with time-varying asymmetric constraints and input saturation. By constructing quartic asymmetric time-varying barrier Lyapunov functions (TVBLFs), all the state variables are not to transgress the prescribed dynamic constraints. The command-filtered backstepping method and the error compensation mechanism are combined to eliminate the issue of "computational explosion" and compensate the filtering errors. An FNN observer is developed to estimate the unmeasured states. The event-triggered mechanism is introduced to improve the efficiency in resource utilization. It is shown that the tracking error can converge to a small neighborhood of the origin, and all signals in the closed-loop systems are bounded. Finally, a physical example is used to verify the feasibility of the theoretical results.
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This brief presents a modified event-triggered command filter backstepping tracking control scheme for a class of uncertain nonlinear systems with unknown input saturation based on the adaptive neural network (NN) technique. First, the virtual control functions are reconstructed to address the uncertainties in subsystems by using command filters. A piecewise continuous function is employed to deal with the unknown input saturation problem. Next, an event-triggered tracking controller is developed by utilizing the adaptive NN technique. Compared with standard NN control schemes based on multiple-function-approximators, our controller only requires a single NN. The closed-loop system stability is analyzed based on the Lyapunov stability theorem, and it is shown that the Zeno behavior is also avoided under the designed event-triggering mechanism. Simulation studies are performed to validate the effectiveness of our controller.