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1.
Food Chem ; 451: 139410, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38670024

RESUMO

Dipicolinic acid (DPA), as a biomarker for Bacillus anthracis, is highly toxic at trace levels. Rapid and on-site quantitative detection of DPA is essential for maintaining food safety and public health. This work develops a dual-channel self-calibrated fluorescence sensor constructed by the YVO4:Eu and Tb-ß-diketone complex for rapid visual detection of DPA. This sensor exhibits high selectivity, fast response time, excellent detection sensitivity, and the detection limit is as low as 4.5 nM in the linear range of 0-16 µM. A smartphone APP and portable ultraviolet lamp can assemble a mobile fluorescence sensor for on-site analysis. Interestingly, adding Cu2+ ions can quench the fluorescence intensity of Tb3+. In contrast, the addition of cysteine can restore the fluorescence, allowing the accurate detection of Cu2+ ions and cysteine in environmental water and food samples. This work provides a portable sensor that facilitates real-time analysis of multiple targets in food and the environment.


Assuntos
Antraz , Bacillus anthracis , Biomarcadores , Cobre , Cisteína , Análise de Alimentos , Contaminação de Alimentos , Ácidos Picolínicos , Smartphone , Cobre/análise , Cisteína/análise , Bacillus anthracis/isolamento & purificação , Bacillus anthracis/química , Biomarcadores/análise , Contaminação de Alimentos/análise , Antraz/diagnóstico , Análise de Alimentos/instrumentação , Análise de Alimentos/métodos , Ácidos Picolínicos/análise , Espectrometria de Fluorescência/instrumentação , Espectrometria de Fluorescência/métodos , Limite de Detecção , Fluorescência , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos
2.
Adv Mater ; 36(5): e2307366, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37805919

RESUMO

"Test-and-go" single-nucleotide variation (SNV) detection within several minutes remains challenging, especially in low-abundance samples, since existing methods face a trade-off between sensitivity and testing speed. Sensitive detection usually relies on complex and time-consuming nucleic acid amplification or sequencing. Here, a graphene field-effect transistor (GFET) platform mediated by Argonaute protein that enables rapid, sensitive, and specific SNV detection is developed. The Argonaute protein provides a nanoscale binding channel to preorganize the DNA probe, accelerating target binding and rapidly recognizing SNVs with single-nucleotide resolution in unamplified tumor-associated microRNA, circulating tumor DNA, virus RNA, and reverse transcribed cDNA when a mismatch occurs in the seed region. An integrated microchip simultaneously detects multiple SNVs in agreement with sequencing results within 5 min, achieving the fastest SNV detection in a "test-and-go" manner without the requirement of nucleic acid extraction, reverse transcription, and amplification.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Nucleotídeos , Proteínas Argonautas , DNA/genética , MicroRNAs/genética , Sondas de DNA
3.
Food Chem ; 439: 138095, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38039616

RESUMO

Excess formaldehyde (FA) is a strong carcinogen, so the development of a rapid visualized and portable formaldehyde detection platform is of great research importance. A multi-color fluorescence sensing system constituted of model compound (NAHN) and red-emitting InP/ZnS QDs was constructed herein, which can simultaneously realize fluorometric-colorimetric dual-mode sensing when exposed to FA environment. Its preparation process was simplified, the detection process was green, and the limits of detection (LOD) were 0.623 µM and 0.791 µM, respectively. The high recoveries of FA in actual water samples indicated that the sensor had broad application prospects. The prepared fluorescent film can be utilized for rapid visual simulation analysis of FA on the surface of various fruits and vegetables. In addition, a serial logic gate was designed to quickly semi-quantitatively assess FA concentration, which promoted the realization of on-site intelligent evaluation of FA.


Assuntos
Colorimetria , Corantes Fluorescentes , Fluorometria , Formaldeído , Limite de Detecção
4.
Artigo em Inglês | MEDLINE | ID: mdl-37917908

RESUMO

Lung cancer, with lung adenocarcinoma comprising over 40% of cases, presents a global health challenge. Evidence indicates that long non-coding RNAs (lncRNAs), such as GUSBP11, could have therapeutic potential. Thus we explored the role and mechanism of GUSBP11 in lung adenocarcinoma. Bioinformatics analyses demonstrated GUSBP11 was upregulated in lung adenocarcinoma and was correlated with worsening prognosis. Quantitative PCR (qPCR) analysis revealed that of GUSBP11 was highly expressed in 61 paired lung adenocarcinoma patient tumor compared to paracancerous tissue samples. GUSBP11 knockdown suppressed lung adenocarcinoma cells proliferation and metastasis in vitro while promoted cell apoptosis, and the silencing of GUSBP11 impaired in vivo tumor growth in lung adenocarcinoma. Mechanistic insights revealed GUSBP11's role in inhibiting the regulatory functions of KHSRP, a protein essential for lung adenocarcinoma cell proliferation and metastasis. Taken together, our findings underscore the therapeutic and diagnostic potential of targeting the GUSBP11-KHSRP axis in lung adenocarcinoma, paving the way for further exploration in clinical settings.

5.
J Nutr Biochem ; 121: 109432, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37657642

RESUMO

Maternal protein malnutrition leads to liver dysfunction and increases susceptibility to nonalcoholic fatty liver disease in adult fetal growth restriction (FGR) offspring, yet the underlying mechanism remains unknown. Peroxisomes play vital roles in fatty acid ß-oxidation (FAO) and detoxification of reactive oxygen species (ROS). Using a well-defined rat model, the peroxins (PEXs), fatty acid metabolic enzymes, and oxidase stress regulators were investigated in the liver of FGR offspring. The results revealed that PEX3, 11b, 14, and 19 were obviously reduced in the fetal liver and lasted to adulthood, suggesting a decrease in the biogenesis and division of peroxisomes. FA metabolism enzymes and ferroptosis regulators were deregulated. To further investigate this association, small interfering RNA was employed to achieve knockdown (KD) of PEX14 in BRL cells (a rat hepatocyte line). PEX14 KD led to dysregulation of PEXs and long-chain FAs accumulation. PEX14 deficiency caused ROS accumulation and lipid peroxidation, finally induced regulated cell death (including apoptosis, autophagy, and ferroptosis). Double knock down (DKD) of PEX14 and fatty acyl-CoA reductase 1 (FAR1) revealed that PEX14 KD-induced ferroptosis was related with enhanced FAR1 level. DKD of PEX14 and Atg5 further confirmed that PEX14 KD-induced cell death was partly autophagy-dependent. Overall, these data demonstrate a vital role for PEX14 in maintaining peroxisome function and liver physiology, and suggest that hepatocyte peroxisome defects partly explain liver dysplasia and lipid metabolism disorders in fetal original liver disease.

6.
Sci Adv ; 9(38): eadj0839, 2023 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-37729411

RESUMO

Precision chemistry demands miniaturized catalytic systems for sophisticated reactions with well-defined pathways. An ideal solution is to construct a nanoreactor system functioning as a chemistry laboratory to execute a full chemical process with molecular precision. However, existing nanoscale catalytic systems fail to in situ control reaction kinetics in a closed-loop manner, lacking the precision toward ultimate reaction efficiency. We find an inter-electrochemical gating effect when operating DNA framework-constructed enzyme cascade nanoreactors on a transistor, enabling in situ closed-loop reaction monitoring and modulation electrically. Therefore, a comprehensive system is developed, encapsulating nanoreactors, analyzers, and modulators, where the gate potential modulates enzyme activity and switches cascade reaction "ON" or "OFF." Such electric field-effect property enhances catalytic efficiency of enzyme by 343.4-fold and enables sensitive sarcosine assay for prostate cancer diagnoses, with a limit of detection five orders of magnitude lower than methodologies in clinical laboratory. By coupling with solid-state electronics, this work provides a perspective to construct intelligent nano-systems for precision chemistry.


Assuntos
Bioensaio , Eletricidade , Masculino , Humanos , Catálise , Inteligência , Nanotecnologia
7.
Funct Plant Biol ; 49(10): 887-897, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35798353

RESUMO

We investigated potassium (K) accumulation characteristics and expression of K metabolism related genes in one high-K variety (ND202) and a common variety (NC89) of tobacco (Nicotiana tabacum L.). Results showed that K accumulation and leaf K content in ND202 were higher than those in NC89. The distribution rate and K accumulation in the leaves of ND202 increased significantly, while the distribution rate in the roots and stems had lower values. In addition, the maximum K accumulation rate and high-speed K accumulation duration in ND202 were found to be better than those in NC89. The expression of NKT1 in the upper and middle leaves of ND202 had an advantage, and the relative expression of NtKC1 and NtTPK1 in both the upper and middle leaves, as well as the roots, was also significantly upregulated. Conversely, the expression of NTRK1 in the lower leaves and roots of ND202 was weaker. ND202 had significantly greater expression levels of NtHAK1 than NC89 in the upper and middle leaves and roots; moreover, the expression of NtKT12 in the upper leaves and roots of ND202 was also higher. In comparison with common varieties, high-K varieties had a stronger ability to absorb and accumulate K. They also possessed higher expression of K+ channel- and transporter-related genes and showed a superior K accumulation rate and longer duration of high-speed K accumulation. Furthermore, K accumulation rate at 40-60days can be suggested as an important reference for the selection of high-K tobacco varieties.


Assuntos
Nicotiana , Potássio , Folhas de Planta/genética , Raízes de Plantas/genética , Potássio/metabolismo , Nicotiana/genética
8.
J Cell Physiol ; 236(6): 4313-4329, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33230845

RESUMO

Hepatic stellate cell (HSC) activation plays an important role in the pathogenesis of liver fibrosis, and epithelial-mesenchymal transition (EMT) is suggested to potentially promote HSC activation. Superoxide dismutase 3 (SOD3) is an extracellular antioxidant defense against oxidative damage. Here, we found downregulation of SOD3 in a mouse model of liver fibrosis induced by carbon tetrachloride (CCl4 ). SOD3 deficiency induced spontaneous liver injury and fibrosis with increased collagen deposition, and further aggravated CCl4 -induced liver injury in mice. Depletion of SOD3 enhanced HSC activation marked by increased α-smooth muscle actin and subsequent collagen synthesis primarily collagen type I in vivo, and promoted transforming growth factor-ß1 (TGF-ß1)-induced HSC activation in vitro. SOD3 deficiency accelerated EMT process in the liver and TGF-ß1-induced EMT of AML12 hepatocytes, as evidenced by loss of E-cadherin and gain of N-cadherin and vimentin. Notably, SOD3 expression and its pro-fibrogenic effect were positively associated with sirtuin 1 (SIRT1) expression. SOD3 deficiency inhibited adenosine monophosphate-activated protein kinase (AMPK) signaling to downregulate SIRT1 expression and thus involving in liver fibrosis. Enforced expression of SIRT1 inhibited SOD3 deficiency-induced HSC activation and EMT, whereas depletion of SIRT1 counteracted the inhibitory effect of SOD3 in vitro. These findings demonstrate that SOD3 deficiency contributes to liver fibrogenesis by promoting HSC activation and EMT process, and suggest a possibility that SOD3 may function through modulating SIRT1 via the AMPK pathway in liver fibrosis.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/enzimologia , Colágeno Tipo I/metabolismo , Transição Epitelial-Mesenquimal , Células Estreladas do Fígado/enzimologia , Cirrose Hepática Experimental/enzimologia , Fígado/enzimologia , Superóxido Dismutase/deficiência , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Tetracloreto de Carbono , Células Cultivadas , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/patologia , Células Estreladas do Fígado/patologia , Fígado/patologia , Cirrose Hepática Experimental/induzido quimicamente , Cirrose Hepática Experimental/genética , Cirrose Hepática Experimental/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transdução de Sinais , Sirtuína 1/metabolismo , Superóxido Dismutase/genética
9.
Zhongguo Zhong Yao Za Zhi ; 41(24): 4556-4563, 2016 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-28936837

RESUMO

Soil microbes are the important indicator of soil quality. For exploring Chuanminshen violaceum planting to microbial effects in tobacco soil, this paper adopted Illumina MiSeq high-throughput sequencing to research the change of bacteria and fungi at the phylum and genus in the soil. The results showed that the Ch. violaceum planting increased the biodiversity of bacteria and fungi. The influence on fungi was greater than that on bacteria. It greatly increased the sequence of fungi, it obtained 32 978 16S rDNA and 32 229 18S rDNA sequence number. There was no change of the top three phylums in bacteria, but the content changed, Proteobacteria and Acidobacteria reduced by 1.73% and 1.4% respectively, and Actinobacteria increased by 0.65%. The advantage phylum Ascomycete in tobacco reduced by 27.99% to be second advantage phylum after Ch. violaceum planting, and the second advantage phylum Basidiomycete increased by 23.69% to become the first dominant fungi. At the genus, Ch. violaceum planting changed the order of dominant genus and the abundance was also changed. Some changed largely such as uncultured Acidobacteriaceae Subgroup-1, Gemmatimonas, Subgroup-2,uncultured Nitrosomonadaceae for bacteria, norank Sordariales, norank Agaricomycetes, Phialophora for fungi. Especially the rotation increased antagonistic microbes and physiological microbes and decreased pathogenic microbes. So the Ch. violaceum planting can improve the microbe community in tobacco soil.


Assuntos
Agricultura/métodos , Apiaceae/crescimento & desenvolvimento , Nicotiana/crescimento & desenvolvimento , Microbiologia do Solo , Apiaceae/microbiologia , Bactérias/classificação , Biodiversidade , Fungos/classificação , RNA Ribossômico 16S , Solo , Nicotiana/microbiologia
10.
Waste Manag Res ; 32(3): 179-85, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24616342

RESUMO

Sewage sludge and industrial steel slag (SS) pose threats of serious pollution to the environment. The experiments aimed to improve the stabilizing effects of heavy metal Ni and Cd morphology in composting sludge. The total Ni and Cd species distribution and chemical forms in the compost sewage sludge were investigated with the use of compost and co-compost with SS, including degradation. The carbon/nitrogen ratio of piles was regulated with the use of sawdust prior to batch aerobic composting experiments. Results indicated that the co-composting with SS and organic matter humification can contribute to the formation of Fe and Mn hydroxides and that the humus colloid significantly changed Ni and Cd species distribution. The decreased content of Ni and Cd in an unstable state inhibited their biological activity. Conclusions were drawn that an SS amount equal to 7% of the dry sludge mass was optimal value to guarantee the lowest amount of Cd in an unstable state, whereas the amount was 14% for Ni.


Assuntos
Cádmio/química , Resíduos Industriais/análise , Níquel/química , Esgotos/química , Solo/química , Aço/química , China , Aço/análise
11.
Zhonghua Xin Xue Guan Bing Za Zhi ; 41(7): 572-6, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-24284184

RESUMO

OBJECTIVE: The types and risk factors of arrhythmia were analyzed on acute coronary syndrome (ACS) patients under the age of 44 years who were hospitalized in Henan province between September 2009 to June 2012. METHODS: Medical records of eligible patients were obtained from the information system of the First Affiliated Hospital of Zhengzhou University teleconsultation information center. Middle aged and elderly ACS patients who were hospitalized at the same period served as controls. Data on arrhythmia types, blood pressure, thyroid disease, respiratory sleep apnea syndrome, smoking history, history of alcohol consumption, eating habits, family history of early-onset arrhythmia, laboratory tests were analyzed. RESULTS: (1) Arrhythmia was detected in 110 out of young ACS patients (55%), which was significantly lower than that in the elderly ACS patients (71.05%, P < 0.01). (2) The top three arrhythmias in young ACS patients were: sinus tachycardia (30.50%), the premature ventricular contractions (19.00%), atrial flutter/atrial fibrillation (16.50%). Incidence of sinus tachycardia, atrial flutter/atrial fibrillation were significantly higher while incidence of ventricular tachycardia, ventricular fibrillation, paroxysmal supraventricular tachycardia were significantly lower in young ACS patients than in middle-aged ACS patients (all P < 0.05). The incidence of sinus tachycardia was higher while incidence of ventricular premature accelerated ventricular spontaneous cardiac rhythm, ventricular tachycardia, ventricular fibrillation, non-paroxysmal supraventricular tachycardia, atrial flutter/atrial fibrillation, paroxysmal supraventricular tachycardia, sinus bradycardia, nodal escape, atrioventricular block were significantly lower in young ACS patients than in elderly ACS patients (all P < 0.05). (3) Body mass index, incidence of smoking, coronary three-vessel disease, drinking, eating salty foods, thyroid dysfunction, sleep apnea were significantly higher in youth ACS patients with arrhythmia than in young ACS patients without arrhythmia (all P < 0. 05). (4) Logistic regression analysis found that number of diseased coronary vessels (OR = 24.293), smoking (OR = 1.112) and alcohol consumption (OR = 1.039) were independent risk factor for developing arrhythmia in young ACS patients from Henan province. CONCLUSIONS: The main types of arrhythmia are sinus tachycardia, premature ventricular contractions, atrial flutter/atrial fibrillation and the major risk factors related to the arrhythmia are number of diseased coronary vessels, smoking and alcohol consumption in young ACS patients from Henan province.


Assuntos
Síndrome Coronariana Aguda/complicações , Arritmias Cardíacas/etiologia , Síndrome Coronariana Aguda/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Arritmias Cardíacas/epidemiologia , China/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco
12.
Shanghai Kou Qiang Yi Xue ; 22(2): 126-31, 2013 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-23708021

RESUMO

PURPOSE: To explore the feasibility of dual release baicalin and rhBMP-2 system for the treatment of periodontal diseases in minipigs. METHODS: Four miniature swines were selected to establish the model of periodontitis and randomly divided them into four groups: Dual release group, hydrogel with baicalin-GMS and rhBMP-2; Single Huang group, hydrogel with baicalin-GMS; BMP group, hydrogel with rhBMP-2; Negative control group, blank hydrogel. All parameters including clinical indicators of periodontitis, the level of IL-1 and TNF-α in the gingival crevicular fluid (GCF) were measured before and 6 weeks after modeling, and 4, 8 weeks after treatment. The data was analyzed using SPSS13.0 software package. The periodontitis model animals were sacrificed at the end of 8 weeks after treatment. The histological changes of periodontal tissues were observed under microscope with HE staining. RESULTS: (1)In all groups, there were significant differences of clinical indicators and levels of IL-1 and TNF-α in the GCF between periodontitis was established and 4, 8 weeks after treatment (P<0.05). Dual release group was significantly superior than that of other three groups (P<0.05). Moreover, all the parameters improved continuously. X-ray showed that bone mineral density and height in the remaining three groups increased compared with the control group, with highest in dual release group. (2)The results of histological observation showed that the inflammation reaction of periodontal tissues in negative control group was more serious than that in other groups 8 weeks after treatment. There was no obvious repair reaction in the control group, but different amount of new bone was seen in other three groups. Among them, dual release group had most obvious repair reaction. CONCLUSIONS: The dual-slow-release chitosan thermosensitive hydrogel system is employable to be used effectively in the regeneration of periodontal defects, which laid a foundation for further clinical use.


Assuntos
Líquido do Sulco Gengival , Periodontite , Regeneração , Animais , Flavonoides , Doenças Periodontais , Periodonto , Suínos , Porco Miniatura , Fator de Necrose Tumoral alfa , Cicatrização
13.
Zhonghua Xin Xue Guan Bing Za Zhi ; 41(12): 1000-5, 2013 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-24524600

RESUMO

OBJECTIVE: To evaluate the effects and clinical prognosis of out-patient department-based smoking cessation services for coronary heart disease (CHD) patients. METHODS: A total of 140 smoking patients diagnosed with coronary heart disease in our cardiovascular department were randomly divided into the intensive smoking cessation clinic follow-up group (intervention group, patients were informed on the importance and methods to quit smoking at the first visit and reminded for that at months interval for 6 months, n = 70) and the conventional treatment group (control group, n = 70). After 6 months, the smoking status, cardiovascular event rates, drug usage, out-patient medical costs and quality of life were compared between the two groups. RESULTS: Age, gender, concomitant diseases, drug usage were similar between the two groups at baseline (all P > 0.05). After 6 months, smoking quit rate [34.2% (24/70) vs. 5.7% (4/70), P < 0.01], drug use rates: lipid-lowering drugs [95.3% (67/70) vs. 80.4% (56/70)], ß blockers [82.4% (57/70) vs. 41.3% (28/70)], and ACEI/ARB [61.4% (43/70) vs. 34.4% (24/70)] were significantly higher in the intervention group than in the control group, while total cardiovascular event rates [21.4% (15/70) vs. 47.1% (33/70), P < 0.01] and out-patient medical costs (3789.3 RMB vs. 4984.2 RMB, P < 0.01) were significantly lower in the intervention group than in the control group. The quality of life scores derived from MYO health survey questionnaire was significantly higher in the intervention group than in the control group (P < 0.01). The top three reasons responsible for continuous smoking for all patients failed to quit smoking were: (1) others smoked more than me and still alive and healthy [90.3% (56/62)]; (2) smoking helped me to keep relaxed and reduce trouble in daily work and life [70.9% (44/62)]; (3) smoking was essential while chatting and drinking with friends [66.1% (41/62)]. The overall satisfactory rate to this smoking cessation program was 42.8% and the satisfactory rate was up to 50.0% by patients. CONCLUSIONS: Intensive outpatient smoking cessation follow-up program can significantly improve the smoking cessation rates, the guideline drug use rate and the quality of life while reduce medical costs for coronary heart disease patients.


Assuntos
Doença das Coronárias , Abandono do Hábito de Fumar/métodos , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pacientes Ambulatoriais
14.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 27(7): 767-9, 2011 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-21722530

RESUMO

AIM: Design and synthesis complementary DNA sequences of 3 pairs of short hairpin structure and a pair of negative control sequence by RNA interference technique and construct and identify a lentiviral interference vector with human BIRC5 gene as target gene. METHODS: The designed and synthesised Single-Stranded primer were annealed to Double-Stranded oligo sequences and subcloned into linear pMAGic lentiviral plasmid vector digested by enzyme Age I and EcoR I. Screening positive clone after transformed into DH5α competent cells and identified by PCR amplification and DNA sequencing. RESULTS: 335 bp straps of positive clone and 298 bp straps of negative clone form PCR amplification production have been obtained after gel electrophoresis, the designed and synthesised sequences have been contained in these clone straps confirmed by the result of DNA sequencing. CONCLUSION: Four pairs of BIRC5 shRNA recombinant lentiviral expression vector were constructed successfully, which laid the foundation for researching the inhibition of BIRC5 siRNA target against tumor cells proliferation, induction apoptosis and gene therapy.


Assuntos
Vetores Genéticos/genética , Proteínas Inibidoras de Apoptose/deficiência , Proteínas Inibidoras de Apoptose/genética , Lentivirus/genética , Engenharia de Proteínas/métodos , RNA Interferente Pequeno/genética , Apoptose/genética , Expressão Gênica , Terapia Genética , Humanos , Reação em Cadeia da Polimerase , Survivina
15.
Hepatobiliary Pancreat Dis Int ; 5(3): 428-31, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16911944

RESUMO

BACKGROUND: Brain-dead donors have been the main sources in organ transplantation. But many studies show that brain-death affects the organ's function after transplantation. This study was undertaken to investigate liver injury after brain-death in rats and the protective effects of N-acetyleysteine (NAC) on liver injury. METHODS: A total of 30 Wistar rats were randomized into 3 groups: normal control group (C), brain-dead group (B), and NAC pretreatment group (N). At 4 hours after the establishment of a brain-dead model, serum was collected to determine the levels of ALT, AST, TNF-alpha and hyaluronic acid (HA). Hepatic tissue was obtained for electron microscopic examination. RESULTS: At 4 hours, the levels of ALT, AST, TNF-alpha, and HA in group N were significantly higher than those in group C, but these parameters were significantly lower than those in group B. Electron microscopy showed activated Kupffer cells, denuded sinusoidal endothelial cells (SECs), and widened fenestration in group B, but eliminated activation of Kupffer cells and intact SECs in group N. CONCLUSION: Brain death can cause liver injury, and N-acetyleysteine can protect the liver from the injury.


Assuntos
Acetilcisteína/farmacologia , Morte Encefálica , Fígado/efeitos dos fármacos , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Fígado/patologia , Fígado/ultraestrutura , Microscopia Eletrônica , Ratos , Ratos Wistar
16.
Hepatobiliary Pancreat Dis Int ; 5(2): 237-41, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16698583

RESUMO

BACKGROUND: Phage display technology has become a vital tool in studies aimed at identifying molecules binding to a specific target. It enables the rapid generation and selection of high affinity, fully human antibody product candidates to essentially any disease target appropriate for antibody therapy. In this study, we prepared the recombinant single-chain fragment variable (ScFv) antibody to hepatitis B virus surface antigen (HBsAg) by the phage display technology for obtaining a virus-targeting mediator. METHODS: mRNA was isolated from B-lymphocytes from a healthy volunteer and converted into cDNA. The fragment variables of heavy and light chain were amplified separately and assembled into ScFv DNA with a specially constructed DNA linker by polymerase chain reaction. The ScFv DNA was ligated into the phagmid vector pCANTAB5E and the ligated sample was transformed into competent E.coli TG1. The transformed cells were infected with M13K07 helper phage to form a human recombinant phage antibody library. The volume and recombinant rate of the library were evaluated by bacterial colony count and restriction analysis. After two rounds of panning with HBsAg, the phage clones displaying ScFv of the antibody were selected by enzyme-linked immunosorbant assay (ELISA) from the enriched phage clones. The antigen binding affinity of the positive clone was detected by competition ELISA. HB2151 E.coli was transfected with the positive phage clone demonstrated by competition ELISA for production of a soluble form of the anti-HBsAg ScFv. ELISA assay was used to detect the antigen binding affinity of the soluble anti-HBsAg ScFv. Finally, the relative molecular mass of soluble anti-HBsAg ScFv was measured by SDS-PAGE. RESULTS: The variable heavy (VH) and variable light (VL) and ScFv DNAs were about 340 bp, 320 bp and 750 bp, respectively. The volume of the library was up to 2 x 10(6) and 8 of 10 random clones were recombinants. Two phage clones could strongly compete with the original HBsAb for binding to HBsAg. Within 2 strong positive phage clones, the soluble anti-HBsAg ScFv from one clone was found to have the binding activity with HBsAg. SDS-PAGE showed that the relative molecular weight of soluble anti-HBsAg ScFv was 32 kDa. CONCLUSION: The anti-HBsAg ScFv successfully produced by phage antibody technology may be useful for broadening the scope of application of the antibody.


Assuntos
Anticorpos Monoclonais/metabolismo , Antígenos de Superfície da Hepatite B/imunologia , Fragmentos de Imunoglobulinas/imunologia , Região Variável de Imunoglobulina/metabolismo , Anticorpos Monoclonais/imunologia , Afinidade de Anticorpos , Ensaio de Imunoadsorção Enzimática , Estudos de Avaliação como Assunto , Humanos , Fragmentos de Imunoglobulinas/metabolismo , Programas de Rastreamento/métodos , Biblioteca de Peptídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estudos de Amostragem , Sensibilidade e Especificidade
17.
World J Gastroenterol ; 11(7): 1023-7, 2005 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-15742407

RESUMO

AIM: To detect the expression of CK20mRNA in peripheral blood of pancreatic cancer and evaluate its clinical significance. METHODS: Expression of CK20mRNA in peripheral blood was detected by fluorogenic qualitative reverse transcription-polymerase chain reaction (RT-PCR) in 40 cases of pancreatic cancer at the night before operation, in 5 cases of benign pancreatic diseases, in 5 cases of healthy individuals. The relationships were investigated between CK20mRNA expression and the clinicopathological variables, and clinical follow-up outcome in those patients with pancreatic cancer having undergone radical resection. RESULTS: Of the 40 patients with pancreatic cancer, 23 (57.5%) cases were positive for CK20mRNA expression. CK20mRNA expression was significantly correlated with lymphatic metastasis (P = 0.008), histopathological grading (P = 0.009), and pathological stage (P = 0.021); there was no significant correlation between CK20mRNA expression and age, gender, tumor diameter, and depth of invasion. The cumulative metastasis rates of patients with CK20mRNA expression were higher than those of patients with no CK20mRNA expression within 6 mo (34.7% vs 5.9%, P = 0.043) or 12 mo (73.9% vs 35.3%, P = 0.02) after operation. CK20mRNA expression in peripheral blood of pancreatic cancer indicated poorer prognosis. The survival rate of patients with CK20mRNA expression was lower than that of patients with negative CK20mRNA expression (Log-Rank = 13.31, P = 0.0003). CONCLUSION: CK20mRNA is a sensitive and specific molecular marker for the detection of micrometastasis in peripheral blood of patients with pancreatic cancer. The CK20mRNA expression in peripheral blood is correlated with biological characteristic of pancreatic cancer. It can help to predict the prognosis of pancreatic cancer after operation, and to determine which patient will benefit from aggressive adjuvant therapies.


Assuntos
Biomarcadores Tumorais/sangue , Proteínas de Filamentos Intermediários/genética , Neoplasias Pancreáticas/sangue , Neoplasias Pancreáticas/secundário , Adulto , Idoso , Feminino , Seguimentos , Humanos , Queratina-20 , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/mortalidade , Neoplasias Pancreáticas/cirurgia , Prognóstico , RNA Mensageiro/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sobrevida
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