RESUMO
Decades of neuroscience research has shown that macroscale brain dynamics can be reliably decomposed into a subset of large-scale functional networks, but the specific spatial topographies of these networks and the names used to describe them can vary across studies. Such discordance has hampered interpretation and convergence of research findings across the field. To address this problem, we have developed the Network Correspondence Toolbox (NCT) to permit researchers to examine and report spatial correspondence between their novel neuroimaging results and sixteen widely used functional brain atlases, consistent with recommended reporting standards developed by the Organization for Human Brain Mapping. The atlases included in the toolbox show some topographical convergence for specific networks, such as those labeled as default or visual. Network naming varies across atlases, particularly for networks spanning frontoparietal association cortices. For this reason, quantitative comparison with multiple atlases is recommended to benchmark novel neuroimaging findings. We provide several exemplar demonstrations using the Human Connectome Project task fMRI results and UK Biobank independent component analysis maps to illustrate how researchers can use the NCT to report their own findings through quantitative evaluation against multiple published atlases. The NCT provides a convenient means for computing Dice coefficients with spin test permutations to determine the magnitude and statistical significance of correspondence among user-defined maps and existing atlas labels. The NCT also includes functionality to incorporate additional atlases in the future. The adoption of the NCT will make it easier for network neuroscience researchers to report their findings in a standardized manner, thus aiding reproducibility and facilitating comparisons between studies to produce interdisciplinary insights.
RESUMO
Objective: To investigate the correlation between serum ferritin (SF) level and liver damage in the acute stage of dengue fever. Methods: A retrospective study was conducted to analyze 171 cases diagnosed with dengue fever as dengue fever group and 130 healthy patients as control group in Hangzhou 3A grade hospital from July to December 2017. Clinical data, SF and liver function related indicators were collected from both groups: alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL) to analyze the correlation between liver damage and SF in patients with dengue fever. Results: ALT, AST, and SF levels were significantly higher in the dengue fever group than those in the healthy control group (Z = 11.553, 15.054 and 15.163, P < 0.001). SF levels were higher in the dengue fever combined with liver damage group than those without the liver damage group (z = 6.930, P < 0.001). However, there was no statistically significant differences in age, gender, peak body temperature, and history of liver disease (P > 0.05). In addition, Spearman's correlation analysis showed that SF was positively correlated with ALT, AST, and TBIL (r = 0.464, 0.531 and 0.315, P < 0.001). Among dengue patients with different SF levels, there were significant difference in ALT, AST levels and incidence of liver damage (H = 14.240 and 17.584, χ(2) = 49.547, P < 0.001). Patients with higher SF levels had higher ALT, AST levels and incidence of liver damage. Binary logistic regression analysis showed that hyperferritinemia (SF≥500 ng/ml) was the risk factor for dengue fever combined with liver damage (OR = 8.120, P < 0.001). Furthermore, ROC curve analysis showed that the AUC for SF to judge dengue fever combined liver damage was 0.846 (95% CI: 0.785-0.908), and the sensitivity and specificity when the SF cut-off value was 1 506 ng/ml were 74.8% and 83.3%. Conclusion: There is a certain correlation between the SF level and the degree of liver damage in acute stage of dengue fever patients, and hyperferritinemia is a risk factor for dengue fever combined with liver damage.
Assuntos
Dengue , Hepatopatias , Alanina Transaminase , Aspartato Aminotransferases , Dengue/complicações , Dengue/epidemiologia , Ferritinas , Humanos , Fígado , Estudos RetrospectivosRESUMO
We report a subharmonic (frequency-divide-by-2) optical parametric oscillator (OPO) with a continuous wavelength span of 3 to 12 µm (-37dB level) that covers most of the molecular rovibrational "signature" region. The key to obtaining such a wide spectral span is the use of an OPO with a minimal dispersion-through the choice of intracavity elements, the use of all gold-coated mirrors, and a special "injector" mirror. The system delivers up to 245 mW of the average power with the conversion efficiency exceeding 20% from a 2.35 µm Kerr-lens mode-locked pump laser.
RESUMO
Opiate addiction has a high rate of relapse. The accumulating evidence shows that electroacupuncture (EA) may be effective for the treatment of opiate relapse. However, the change of expression of CB1-Rs and CB2-Rs involve in 2Hz EA anti-relapse pathway is still unclear. To explore the changes of expression of CB1-Rs and CB2-Rs, heroin self-administration (SA) model rats were adopted and treated using 2Hz EA. The expressions of CB1-Rs and CB2-Rs were observed using immunohistochemistry method. The results showed that, compared with the control group, active pokes in the heroin-addicted group increased, while the active pokes decreased significantly in 2Hz EA group compared with heroin-addicted group. Correspondingly, the expression of CB1-Rs in prefrontal cortex (PFC), hippocampus (Hip), nucleus accumbens (NAc) and ventral tegmental area (VTA) all increased significantly while the expression of CB2-Rs in those relapse-relevant brain regions decreased obviously in heroin-addicted group when compared with the control group. In addition, the expression of CB1-Rs obviously decreased in the 2Hz EA group while the expression of CB2-Rs in those relapse-relevant brain regions increased significantly when compared with the heroin-addicted group. It indicated that 2Hz EA could attenuate the heroin-evoked seeking behaviors effectively. The anti-relapse effects of 2Hz EA might be related to the decrease of CB1-Rs and increase of CB2-Rs expression in relapse-relevant brain regions of heroin SA rats.
Assuntos
Comportamento Animal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Comportamento de Procura de Droga/efeitos dos fármacos , Eletroacupuntura , Dependência de Heroína/terapia , Heroína/administração & dosagem , Antagonistas de Entorpecentes/administração & dosagem , Receptor CB1 de Canabinoide/metabolismo , Receptor CB2 de Canabinoide/metabolismo , Animais , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Modelos Animais de Doenças , Extinção Psicológica/efeitos dos fármacos , Dependência de Heroína/metabolismo , Dependência de Heroína/fisiopatologia , Dependência de Heroína/psicologia , Locomoção/efeitos dos fármacos , Masculino , Ratos Sprague-Dawley , Recidiva , Autoadministração , Transdução de SinaisRESUMO
The hyperpolarization-activated cyclic-nucleotide-gated non-selective cation (HCN) channels play a potential role in the neurological basis underlying drug addiction. However, little is known about the role of HCN channels in methamphetamine (METH) abuse. In the present study, we examined the changes in working memory functions of METH re-exposed mice through Morris water maze test, and investigated the protein expression of HCN1 channels and potential mechanisms underlying the modulation of HCN channels by Western blotting analysis. Mice were injected with METH (1 mg/kg, i.p.) once per day for 6 consecutive days. After 5 days without METH, mice were re-exposed to METH at the same concentration. We found that METH re-exposure caused an enhancement of working memory, and a decrease in the HCN1 channels protein expression in both hippocampus and prefrontal cortex. The phosphorylated extracellular regulated protein kinase 1/2 (p-ERK1/2), an important regulator of HCN channels, was also obviously reduced in hippocampus and prefrontal cortex of mice with METH re-exposure. Meanwhile, acute METH exposure did not affect the working memory function and the protein expressions of HCN1 channels and p-ERK1/2. Overall, our data firstly showed the aberrant protein expression of HCN1 channels in METH re-exposed mice with enhanced working memory, which was probably related to the down-regulation of p-ERK1/2 protein expression.
Assuntos
Regulação para Baixo/fisiologia , Hipocampo/metabolismo , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/biossíntese , Memória de Curto Prazo/fisiologia , Metanfetamina/toxicidade , Canais de Potássio/biossíntese , Córtex Pré-Frontal/metabolismo , Animais , Regulação para Baixo/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/antagonistas & inibidores , Locomoção/efeitos dos fármacos , Locomoção/fisiologia , Masculino , Memória de Curto Prazo/efeitos dos fármacos , Metanfetamina/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Córtex Pré-Frontal/efeitos dos fármacos , Distribuição AleatóriaRESUMO
The recombinant sTRAIL has been in clinical trial for various human malignancies. However, the half-life time of sTRAIL is very short, which might be an important factor influencing its clinical efficacy for cancer therapy. We previously reported the recombinant adeno-associated virus (AAV)-encoding sTRAIL95-281-mediated sTRAIL expression in vivo up to 8 months and suppressed tumor growth markedly in mouse xenografts. In the present study, we further evaluated the clinical potency for cancer gene therapy and the safety in mouse and non-human primates. The mouse models with HCT-116, NCI-H460 and BEL-7402 cancers were injected intraperitoneally with a single dose of 1.0 × 1011, 1.0 × 1010 and 1.0 × 109 vg of rAAV2-sTRAIL95-281 virus, respectively. The cynomolgus monkeys were injected (i.m.) with a single dose of rAAV2-sTRAIL95-281 of 1 × 1011, 3 × 1011 and 1 × 1012 vg, corresponding to 6-, 20- and 60-fold of intended use dosage for humans, respectively. The efficacy, pharmacology and toxicity of rAAV-sTRAIL in the animals were analyzed accordingly. The tumor inhibitory rates reached 44-76%, 48-52% and 55-74% in the three tumor models, respectively, and they had no influence on mouse spontaneous activity. Administration (s.c.) of a single dose of rAAV2-sTRAIL95-281 virus of 1.0 × 109 or 1.0 × 1010 vg in mice with implanted tumor led to mainly distribution in the spleen, liver, implanted tumor, blood, injected site of muscle and bone marrow. Two weeks later, there was no rAAV2-sTRAIL95-281 detected in blood and bone marrow, and it significantly decreased in other tissues and organs and then gradually cleared away in 4-12 weeks after administration. There was no rAAV2-sTRAIL accumulation in the animal's body and no influence on the body weights. Administration (i.v.) did not cause animal death, and no dose-related abnormal clinical symptoms were found in the mice. There were no abnormal tissue and organ found in all animals. Long-term toxicity test in cynomolgus monkeys did not cause rAAV2-sTRAIL95-281-related toxic and side effects, except that anti-AAV and anti-sTRAIL antibodies were generated. In conclusion, these data demonstrated that administration of rAAV2-sTRAIL95-281 in mice and in cynomolgus monkeys is safe without obvious toxic and side effects to the animals, and throw light on pharmacokinetics and safety in human clinical trials for cancer gene therapy.
Assuntos
Terapia Genética/efeitos adversos , Neoplasias/genética , Proteínas Recombinantes/administração & dosagem , Ligante Indutor de Apoptose Relacionado a TNF/administração & dosagem , Animais , Dependovirus/genética , Modelos Animais de Doenças , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Vetores Genéticos , Humanos , Macaca fascicularis , Neoplasias/patologia , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/genética , Ligante Indutor de Apoptose Relacionado a TNF/efeitos adversos , Ligante Indutor de Apoptose Relacionado a TNF/genética , Distribuição Tecidual/efeitos dos fármacosRESUMO
BACKGROUND: This paper describes a new multicountry collaborative project to assess the impact of alcohol control policy. Longitudinal surveys of drinkers in a number of participating countries and analysis of the policy context allow for the assessment of change over time within countries and comparison between countries. The design of the study is modeled on the International Tobacco Control study and aims to assess the impact of alcohol policies in different cultural contexts on policy-related behaviors and alcohol consumption. A survey instrument and protocol for policy analysis have been developed by the initial participating countries: England, Scotland, Thailand, South Korea, and New Zealand. The first round of data collection is scheduled for 2011-2012. MEASUREMENTS: The survey instrument (International Alcohol Control [IAC] survey) measures key policy relevant behaviors: place and time of purchase, amounts purchased and price paid; ease of access to alcohol purchase; alcohol marketing measures; social supply; perceptions of alcohol affordability and availability and salience of price; perceptions of enforcement; people's experiences with specific alcohol restrictions; support for policy and consumption (typical quantity, frequency using beverage and location-specific measures). The Policy Analysis Protocol (PoLAP) assesses relevant aspects of the policy environment including regulation and implementation. RESULTS: It has proved feasible to design instruments to collect detailed data on behaviors relevant to alcohol policy change and to assess the policy environment in different cultural settings. CONCLUSIONS: In a policy arena in which the interest groups and stakeholders have different perceptions of appropriate policy responses to alcohol-related harm, a robust methodology to assess the impact of policy will contribute to the debate.
Assuntos
Consumo de Bebidas Alcoólicas/legislação & jurisprudência , Alcoolismo/prevenção & controle , Consumo de Bebidas Alcoólicas/economia , Consumo de Bebidas Alcoólicas/epidemiologia , Custos e Análise de Custo , Estudos Transversais , Coleta de Dados , Inquéritos Epidemiológicos , Humanos , Cooperação Internacional , Estudos Longitudinais , Nova Zelândia , Pesquisa , Inquéritos e QuestionáriosRESUMO
AIM: There is a lack of research, internationally and in New Zealand, on the harms experienced as a result of drinking by others. Such effects have often been neglected in policy development and in estimates of the economic burden associated with alcohol consumption. This study describes the broad range of harms reported by New Zealanders due to the drinking of someone else. METHOD: A representative national survey was conducted using Computer Assisted Telephone Interviewing with New Zealanders aged 12 to 80 years (N=3068) in 2008/2009 (response rate - 64%). Harms experienced due to the drinking of others were reported along with demographic variables. RESULTS: One in four respondents indicated that they had at least one heavy drinker in their life. Most of these respondents indicated they had experienced a range of harms because of this person's drinking. Further, 17% of respondents with children reported that their children experienced harm because of the drinking of someone else. Seventy-one percent of those sampled reported experiencing at least one harm because of the drinking of a stranger. CONCLUSION: A large proportion of New Zealanders report the experience of physical, social, economic, and psychological harms because of the drinking of others. These harms should be considered in the discussion of alcohol policy.
Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Consumo de Bebidas Alcoólicas/epidemiologia , Efeitos Psicossociais da Doença , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Intoxicação Alcoólica/epidemiologia , Intoxicação Alcoólica/psicologia , Criança , Saúde da Família , Medo , Feminino , Amigos , Humanos , Relações Interpessoais , Masculino , Pessoa de Meia-Idade , Nova Zelândia/epidemiologia , Fatores Sexuais , Inquéritos e Questionários , Violência , Local de Trabalho , Adulto JovemRESUMO
In capillary electrophoresis and capillary electrochromatography, the driving factor of the separation is electroosmotic flow (EOF). Pressurized capillary electrochromatography, in which the separation is controlled by EOF as well as the pressure, becomes more and more attractive. We studied the influence of various pressures on capillary electrochromatographic separation. The results reveal that in pressurized capillary electrochromatography, which was performed by EOF combined with the forward and reverse pressure, the main driving factor is still the EOF. It was also found that, when reverse pressure was applied in capillary electrochromatography, the repeatability of the capillary electrochromatographic separation was increased dramatically.
Assuntos
Cromatografia Capilar Eletrocinética Micelar/métodos , Pressão , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To establish a rapid and simple method with high efficiency in detecting point mutation of genomic DNA. METHODS: Four DNA fragments that were different from each other in only one based were amplified by using primers with artificial point mutation based on the sequence of exon 7 of p53 gene, and then were separated by capillary electrophoresis(CE). The neutral coated capillary and 4% linear polyacrylamide gel buffer were used, and the wave length of ultraviolet detector was 254 nm. RESULTS: A homozygous 196 bp DNA fragment can be separated into one dsDNA peak and two ssDNA peaks within 25 minutes, and the heterozygous 196 bp DNA fragments that were made with mixed wild type and mutated DNA can be separated into one dsDNA peak and three ssDNA peaks. The three ssDNA fragments that differ in one nucleotide can be easily separated with good resolution. CONCLUSION: CE technique is rapid, sensitive, accurate and well reproducible. It is an efficient and reliable method for rapidly screening point mutation.
Assuntos
Análise Mutacional de DNA/métodos , DNA/análise , Eletroforese Capilar/métodos , Mutação Puntual , Polimorfismo Conformacional de Fita Simples , Sequência de Bases , DNA/química , Fragmentação do DNA , Primers do DNA , Conformação de Ácido NucleicoRESUMO
On the basis of structure analysis and computer modeling of proUK, two-chain DNA fragment encoding RGD peptide was inserted into the corresponding proUK cDNA site between Gly118-Leu119, using site-directed mutagenesis and DNA recombinant techniques. The chimeric gene was expressed in methylotrophic yeast Pichia pastoris expression system. The chimeric protein was purified after two step purification of Zn2+ chelating column and SP cation exchange column. The specific activity was 65,000 IU/mg protein. The chimeric protein had somewhat lower catalytic efficiency (kcat/Km) on the substrate S2444 as compared to Urokinase. But it had high anti-platelet aggregation activity, and the half inhibit constant was 2.1 mumol/L. The results showed that the chimeric protein not only had higher thrombolytic activity but also obtained anti-thrombus function. Further evaluation of the thrombolytic and antithrombolytic potential in appropriate animal models seemed to be investigated.
Assuntos
Oligopeptídeos/genética , Proteínas Recombinantes/genética , Ativador de Plasminogênio Tipo Uroquinase/genética , Sequência de Aminoácidos , Western Blotting , DNA Recombinante/genética , Relação Dose-Resposta a Droga , Eletroforese em Gel de Poliacrilamida , Expressão Gênica , Humanos , Pichia/genética , Agregação Plaquetária/efeitos dos fármacos , Inibidores da Agregação Plaquetária/farmacologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/farmacologiaRESUMO
Apoptosis is a distinct mode of cell death that is responsible for deletion of cells in normal tissues; it also occurs in specific pathologic contexts. The observation of apoptosis is very important in the research of cancer and cancer therapy. The traditional observation method of apoptosis was agarose gel electrophoresis, which is depending on the determination of ladder-liking DNA fragments extracted from apoptotic cells. It is time-consuming and low-sensitive. Recently, the sieving capillary electrophoresis has been used to detect apoptosis too. However, the problem of DNA fragments contamination is still existing. Here, we have developed a capillary electrophoresis method that could detect apoptosis of whole cell directly and do not need to extract DNA fragments from cells. Apoptosis of adherent cell HeLa cell of carcinoma induced by cyclophosphamide was used as the model to establish the method. The effluence of medicine concentration on apoptosis of cells was studied in detail. It was also found that the method could detect the change of cells in the early period of apoptosis. The induction of apoptosis of HeLa cell by trichosanthin was determined with the method, and the result of flow cytometry was also proved that trichosanthin could result in apoptosis of HeLa cells.
Assuntos
Apoptose/efeitos dos fármacos , Eletroforese Capilar/métodos , Tricosantina/farmacologia , Ciclofosfamida/farmacologia , Células HeLa , HumanosRESUMO
The polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) technique is developed for the detection of point mutations in DNA samples, and is very useful in the research of tumors. The traditional SSCP was carried out with slab gel electrophoresis (SGE), but this is time-consuming and labor-intensive, particularly for clinical diagnoses. We have developed a capillary electrophoresis (CE) method for SSCP detection with a linear polyacrylamide gel solution as the sieving matrix. Twenty colon tumor samples were detected with SSCP-CE and the point mutation in exon 7 of the p53 gene was found in six of the samples. Based on the sequencing results, the accuracy of SSCP-CE was better than that of SSCP-SGE. We hope this rapid and convenient method could be applied in the clinical diagnosis of tumors soon.
Assuntos
Neoplasias do Colo/genética , Eletroforese Capilar/métodos , Genes p53 , Mutação Puntual , Polimorfismo Conformacional de Fita Simples , Sequência de Bases , Primers do DNA , HumanosRESUMO
A pressurized gradient capillary electrochromatography (pCEC) instrument was developed to separate 18 amino acid derivatives. A reversed-phase C18 column (3 microm, 130 mm x 75 microm I.D.) and an acetate buffer (50 mmol/l NaAc, pH 6.4) with an ion-pair reagent (1% N,N-dimethylformamide) were used to separate derivatized amino acids from a standard solution (2 microg/ml), and the wavelength of the UV-Vis detector was 360 nm. The pressure on the capillary column was kept at approx. 70 Pa and 3 kV positive voltage was added on the outlet end of column. The effect of voltage on the eluting order of amino acids and the resolution of separation were studied, and it was found that when the voltage was higher than 3 kV, the adsorption of amino acids in the porous C18 column occurred. The effect of salt concentration, injection volume, and column length on the separation of amino acids was determined. The amino acid sample was separated by pCEC, and RSDs of the migration times of each amino acid were all less than 2.5%.
Assuntos
Aminoácidos/isolamento & purificação , Cromatografia Líquida de Alta Pressão/métodos , Eletroforese Capilar/métodos , Pressão , Espectrofotometria UltravioletaRESUMO
Previous findings have indicated that HIV-1 gp41 like human type I interferon (IFN) could inhibit lymphocyte proliferation and up-modulate MHC class I, II and ICAM-1 molecule expression, and a common epitope exists between gp41 and type I interferon (IFN-alpha and -beta) in the receptor binding regions. To clarify the relationship between human type I interferon and HIV-1 gp41, we tried to inhibit recombinant soluble gp41-binding to human T, B and monocyte cell lines by human IFN-alpha, -beta and -gamma. It was interestingly observed that IFN-beta after preincubating with cells could inhibit the binding of rsgp41 to H9, Raji and U937 cells (T, B and monocyte cell lines), while this binding could not be inhibited by another type I interferon (IFN-alpha) and a type II interferon (IFN-gamma). It was further examined whether human IFN-alpha and -beta bind to the gp41 binding protein P50. In ELISA-assay, the human IFN-beta, but not IFN-alpha, could bind to P50 which was identified as a potential cellular receptor protein for gp41-binding. By the affinity capillary electrophoresis (ACE) analysis, formation of stable IFN-beta-P50 complex was observed. These results indicate that IFN-beta binds the potential receptor protein P50. Based on these experimental evidences and previous studies, it was presumed that the potential cellular receptor protein P50 may be the 51 kDa subunit of human IFN-alpha/beta receptor, which needs to be verified in the future.
Assuntos
Proteína gp41 do Envelope de HIV/metabolismo , Interferon beta/fisiologia , Linfócitos/metabolismo , Monócitos/metabolismo , Receptores de HIV/metabolismo , Linhagem Celular , Eletroforese Capilar , Epitopos/metabolismo , Antígenos HIV/imunologia , Antígenos HIV/metabolismo , Proteína gp41 do Envelope de HIV/imunologia , Humanos , Interferon Tipo I/imunologia , Interferon Tipo I/metabolismo , Interferon-alfa/metabolismo , Interferon beta/metabolismo , Interferon gama/metabolismo , Isoformas de Proteínas/metabolismo , Isoformas de Proteínas/fisiologia , Proteínas RecombinantesRESUMO
The phospholipid monolayer at an air/water interface is widely used to mimic the biological membrane. The dynamic process of the protein or peptide interacting with lipid molecules can be reflected in the change in surface pressure of the monolayer. But the conventional method used to measure the surface pressure change gives results that cannot easily be correlated with the contribution of a single protein molecule. Previously, measuring the surface concentration of the protein molecules at the air/water interface has required the protein to be labeled with radioactivity or fluorescence. Here, a new method using capillary electrophoresis is introduced to measure the surface concentration of the protein. The results show at least two advantages of the new method: The numerical results of protein concentration can be obtained in a more precise and rapid way; and there is no need to label the protein sample or to build a special monolayer setup.
Assuntos
Membrana Celular/metabolismo , Eletroforese Capilar/métodos , Glicoproteínas/biossíntese , Dimiristoilfosfatidilcolina/metabolismo , Humanos , Indicadores e Reagentes/metabolismo , Membranas Artificiais , Fosfatidilgliceróis/metabolismo , Fosfolipídeos/metabolismo , Fatores de Tempo , beta 2-Glicoproteína IRESUMO
Based on the separation of 1-palmitoyl-2-(13-hydroperoxy-cis-9,trans-11-octadecadienoyl)-L-3- phosphatidylcholine (PC-OOH) and 1-palmitoyl-2-(13-hydroxy-cis-9,trans-11-octadecadienoyl)-L-3- phosphatidylcholine (PC-OH) and the quantitative determination of PC-OH, the enzymatic activity of phospholipid hydroperoxide glutathione peroxidase (PHGPx) can be measured by capillary electrophoresis. The separation was carried out in a fused-silica capillary (30 cm x 100 microns id) at 15 kV positive voltage. Sodium borate (100 mM; pH = 8.4) was used as the running buffer, and the photodiode array detector wavelength was 232 nm. The determination can be completed in 5 min. The detection limit was 5 pmol; and the relative standard deviation (RSD) of the peak area was less than 1% with an average recovery of 98.6%. Compared with traditional methods such as HPLC and spectrophotometry, it is faster and more convenient. Using capillary electrophoresis, the enzymatic activities of PHGPx expressed by the rice PHGPx gene in E. coli. M15 was determined as 1.25 x 10(-5) mumol min-1, and the specific activity of partially purified trans-gene PHGPx was 3.1 x 10(-2) mumol min-1 per mg. The stability of the trans-gene PHGPx was also studied.
Assuntos
Glutationa Peroxidase/metabolismo , Calibragem , Eletroforese Capilar , Escherichia coli/enzimologia , Glutationa Peroxidase/genética , Oryza/genética , Fosfolipídeo Hidroperóxido Glutationa Peroxidase , TransgenesRESUMO
Capillary isoelectric focusing electrophoresis (cIEF) has been developed to detect the genotype of apolipoprotein H, which was purified from the serum of a Chinese subject. Depending upon the observation of splitting peaks in cIEF, it is possible to determine if the protein was expressed from a heterozygote gene.
Assuntos
Eletroforese das Proteínas Sanguíneas/métodos , Eletroforese Capilar/métodos , Triagem de Portadores Genéticos/métodos , Glicoproteínas/genética , Focalização Isoelétrica/métodos , Genótipo , Glicoproteínas/isolamento & purificação , Humanos , beta 2-Glicoproteína IRESUMO
OBJECTIVE: Exploring the relationship between Liver-Blood Stasis (LBS) and liver fibrosis (LF) to lay a theoretical foundation of rational using traditional Chinese medicines against LF. METHODS: Human procollagen peptide III (hPC III), hyaluronic acid (HA), laminin (LN) in the sera of 35 patients with hepatopathy and Blood Stasis Syndrome (HP-BS) and 35 patients with hepatopathy and Non-Blood Stasis Syndrome (HP-NBS) were measured by radioimmunoassay. Thirty healthy subjects were taken as control. Correlation analysis between the degrees of LBS and those of the serum indexes of LF was made. RESULTS: (1) hPC III, HA, LN in the sera of the patients with HP-BS were markedly higher than those in the sera of the patients with HP-NBS, but the latter were markedly higher than healthy subjects. (2) Degrees of LBS correlated closely with those of LF. (3) (Xuefu Zhuyu Decoction XFZYD) not only might improve the degrees of LBS but also decline the serum LF indexes. Nevertheless, the curative effects in early period of taking the herbs only showed serum HA dropped. CONCLUSION: The nature of LF in traditional Chinese medicine concept is mainly LBS. The degrees of LBS might reflect those of LF to a certain extent. XFZYD showed effective in declining serum LF indexes but it needs at least 2 months.