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1.
BMC Musculoskelet Disord ; 25(1): 656, 2024 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-39169286

RESUMO

OBJECTIVE: To investigate the clinical significance of using 3D printing guides in modified unilateral puncture percutaneous vertebroplasty (PVP) for the treatment of osteoporotic vertebral compression fractures (OVCF), and to explore a new method for preventing paravertebral vein leakage during PVP in conjunction with a previous study of the optimal puncture-side bone cement/vertebral volume ratio(PSBCV/VV%). METHODS: This retrospective study analyzed 99 patients who underwent unilateral puncture PVP between January 2023 and December 2023. Patients were divided into a guide plate group (46 patients) and a conventional group (53 patients). The guide plate group underwent modified unilateral puncture PVP with the guidance of 3D printing guides, while the conventional group underwent unilateral puncture PVP using the conventional pedicle approach. The distribution of bone cement, surgical outcomes, and the occurrence of cement leakage into paravertebral veins were observed in both groups. RESULTS: The guide plate group had significantly shorter operating time and required fewer fluoroscopies compared to the conventional group. The amount of bone cement volume (BCV) used in the guide plate group was higher, but the amount of bone cement volume on the puncture side(PSBCV), the PSBCV/VV%, and the rate of paravertebral vein leakage were lower in the guide plate group compared to the conventional group (P < 0.05). Within each group, significant improvements in anterior vertebral margin height, Cobb angle, visual analog scale (VAS) score, and Oswestry Disability Index (ODI) were observed at 1 day and 1 month postoperatively compared to preoperative values (P < 0.05). CONCLUSION: Using 3D printing guides in modified unilateral puncture PVP is a safe and effective method for treating OVCF. And it has the advantages of short operation time, less fluoroscopy, even distribution of bone cement, and a low rate of paravertebral vein leakage.


Assuntos
Cimentos Ósseos , Fraturas por Compressão , Fraturas por Osteoporose , Impressão Tridimensional , Fraturas da Coluna Vertebral , Vertebroplastia , Humanos , Estudos Retrospectivos , Fraturas por Compressão/cirurgia , Fraturas por Compressão/diagnóstico por imagem , Feminino , Vertebroplastia/métodos , Masculino , Idoso , Fraturas por Osteoporose/cirurgia , Fraturas da Coluna Vertebral/cirurgia , Fraturas da Coluna Vertebral/diagnóstico por imagem , Pessoa de Meia-Idade , Idoso de 80 Anos ou mais , Cimentos Ósseos/uso terapêutico , Resultado do Tratamento , Punções/métodos , Relevância Clínica
2.
Curr Issues Mol Biol ; 46(7): 6769-6782, 2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-39057046

RESUMO

Camellia fascicularis has important ornamental, medicinal, and food value. It also has tremendous potential for exploiting bioactivities. However, the bioactivities of secondary metabolites in C. fascicularis have not been reported. The structures of compounds were determined by spectral analysis and nuclear magnetic resonance (NMR) combined with the available literature on secondary metabolites of C. fascicularis leaves. In this study, 15 compounds were identified, including 5 flavonoids (1-5), a galactosylglycerol derivative (6), a terpenoid (7), 4 lignans (8-11), and 4 phenolic acids (12-15). Compounds 6-7 and 9-12 were isolated from the genus Camellia for the first time. The remaining compounds were also isolated from C. fascicularis for the first time. Evaluation of antioxidant and antimicrobial activities revealed that compounds 5 and 8-11 exhibited stronger antioxidant activity than the positive drug ascorbic acid, while compounds 7, 13, and 15 showed similar activity to ascorbic acid. The minimum inhibitory concentration (MIC) of antibacterial activity for compounds 5, 7, 9, 11, and 13 against Pseudomonas aeruginosa was comparable to that of the positive control drug tetracycline at a concentration of 62.50 µg/mL; other secondary metabolites inhibited Escherichia coli and Staphylococcus aureus at concentrations ranging from 125-250 µg/mL.

3.
Foods ; 13(14)2024 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-39063349

RESUMO

Camellia fascicularis has important ornamental, medicinal, and food values, which also have tremendous potential for exploiting bioactivities. We performed the bioactivity-guided (antioxidant and antimicrobial) screening of eight fractions obtained from the ethyl acetate phase of C. fascicularis. The antioxidant activity was measured by DPPH, ABTS, and FRAP, and the antibacterial activity was measured by the minimum inhibitory concentration (MIC) of Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus. The results of bioactivity-guided isolation indicated that the major antioxidant compounds in the ethanolic extracts of C. fascicularis may be present in fractions (Fr.) (A-G, obtained after silica gel column chromatography). Fr. (D-I, obtained after silica gel column chromatography) is a fraction of C. fascicularis with antimicrobial activity. The structures of compounds were determined by spectral analysis and nuclear magnetic resonance (NMR) combined with the available literature on secondary metabolites of C. fascicularis leaves. In this study, 17 compounds were identified, including four phenolics (1, 3-4, and 14), a phenylpropane (2), five terpenoids (5-7, 12, and 15), four flavonoids and flavonoid glycosides (8-10 and 16), and two lignins (13 and 17). Compounds 4-7, 13-15, and 17 were isolated from the genus Camellia for first time. The remaining compounds were also isolated from C. fascicularis for first time. The evaluation of antioxidant and antimicrobial activities revealed that compounds 1, 3, 9, 11, and 17 exhibited higher antioxidant activity than the positive control drug (ascorbic acid), and compounds 4, 8, 10, and 13 showed similar activity to ascorbic acid. The other compounds had weaker or no significant antioxidant activities. The MIC of antibacterial activity for compounds 4, 7, and 11-13 against P. aeruginosa was comparable to that of the positive control drug tetracycline at 125 µg/mL, and other secondary metabolites inhibited E. coli and S. aureus at 250-500 µg/mL. This is also the first report of antioxidant and antimicrobial activities of compounds 5-7, 13-15, and 17. The results of the study enriched the variety of secondary metabolites of C. fascicularis and laid the foundation for further research on the pharmacological efficacy and biological activity of this plant.

4.
Cell Death Discov ; 10(1): 212, 2024 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-38697996

RESUMO

The escalating threat of bone-related diseases poses a significant challenge to human health. Mesenchymal stem cell (MSC)-derived extracellular vesicles (MSC-EVs), as inherent cell-secreted natural products, have emerged as promising treatments for bone-related diseases. Leveraging outstanding features such as high biocompatibility, low immunogenicity, superior biological barrier penetration, and extended circulating half-life, MSC-EVs serve as potent carriers for microRNAs (miRNAs), long no-code RNAs (lncRNAs), and other biomolecules. These cargo molecules play pivotal roles in orchestrating bone metabolism and vascularity through diverse mechanisms, thereby contributing to the amelioration of bone diseases. Additionally, engineering modifications enhance the bone-targeting ability of MSC-EVs, mitigating systemic side effects and bolstering their clinical translational potential. This review comprehensively explores the mechanisms through which MSC-EVs regulate bone-related disease progression. It delves into the therapeutic potential of MSC-EVs as adept drug carriers, augmented by engineered modification strategies tailored for osteoarthritis (OA), rheumatoid arthritis (RA), osteoporosis, and osteosarcoma. In conclusion, the exceptional promise exhibited by MSC-EVs positions them as an excellent solution with considerable translational applications in clinical orthopedics.

6.
Kaohsiung J Med Sci ; 37(6): 459-467, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33611825

RESUMO

Dysregulation of the long intergenic noncoding RNA 01315 (LINC01315) has recently been demonstrated in cancer. However, the role of LINC01315 in papillary thyroid cancer (PTC) has not been determined. We attempted to determine the function of LINC01315 in PTC. The levels of LINC01315 were higher in thyroid carcinoma tissues and cell lines compared with that in noncancerous tissues or normal cells, respectively. LINC01315 knockdown significantly inhibited the in vitro colony formation and invasion of PTC cells. Upregulation of LINC01315 produced opposite effects. Bioinformatic analysis and luciferase reporter assays indicated direct binding of miR-497-5p to LINC01315. Gain- and loss-of-function assays indicated that miR-497-5p acts as a suppressive miRNA in PTC. Furthermore, LINC01315 facilitated the growth and invasion of PTC cells by sponging miR-497-5p. Our results demonstrated the critical role of the LINC01315-miR-497-5p axis in the growth and invasion of PTC cells.


Assuntos
RNA Longo não Codificante/farmacologia , Câncer Papilífero da Tireoide/genética , Câncer Papilífero da Tireoide/metabolismo , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/metabolismo , Adulto , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Biologia Computacional , Feminino , Humanos , Luciferases/metabolismo , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Invasividade Neoplásica , Fenótipo , Transfecção , Regulação para Cima
7.
Waste Manag ; 103: 370-377, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31927327

RESUMO

To understand the evolution and mechanism of the hydrothermal carbonization products of colored cotton fiber, hydrothermal carbonization experiments were carried out on colored cotton fiber dyed with reactive red X-3B and reactive blue KN-R. Fourier transform infrared spectroscopy, X-ray diffraction, and elemental analysis were used to characterize the hydrochar from colored cotton fiber and the hydrothermal behavior of reactive dyes under different reaction temperatures and time conditions. The study showed that when the reaction temperature was in the range of 210-250 °C, hydrolysis of reactive dyes was complete in 6 h, and organic acid that is produced by the degradation of cotton fiber accelerates the hydrolysis of dyes. Also, the hydrolysis products of dyes were involved in the hydrothermal carbonization of cotton fiber and covered the inside of the carbon microspheres; this led to changes in the elemental composition and functional group of the hydrochar. However, the dye content in colored fabric was limited because of limited dye-uptake. Consequently, the reactive activity of dyes was not strong enough to change the hydrothermal behavior of cotton fiber.


Assuntos
Corantes , Fibra de Algodão , Carbono , Hidrólise , Temperatura
8.
Front Plant Sci ; 10: 1270, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31649715

RESUMO

To identify drought-tolerant crop cultivars or achieve a balance between water use and yield, accurate measurements of crop water stress are needed. In this study, the canopy temperature (Tc) of maize at the late vegetative stage was extracted from high-resolution red-green-blue (RGB, 1.25 cm) and thermal (7.8 cm) images taken by an unmanned aerial vehicle (UAV). To reduce the number of parameters for crop water stress monitoring, four simple methods that require only Tc were identified: Tc, degrees above non-stress, standard deviation of Tc, and variation coefficient of Tc. The ground-truth temperatures obtained using a handheld infrared thermometer were used to calibrate the temperature obtained from the UAV thermal images and to evaluate the Tc extraction results. Measured leaf stomatal conductance values were used to evaluate the performance of the four Tc-based crop water stress indicators. The results showed a strong correlation between ground-truth Tc and Tc extracted by the red-green ratio index (RGRI)-Otsu method proposed in this study, with a coefficient of determination of 0.94 (n = 15) and root mean square error value of 0.7°C. The RGRI-Otsu method was most accurate for estimating temperatures around 32.9°C, but the magnitude of residuals increased above and below this value. This phenomenon may be attributable to changes in canopy cover (leaf curling) under water stress, resulting in changes in the proportion of exposed sunlit soil in UAV thermal orthophotographs. Therefore, to improve the accuracy of maize canopy detection and extraction, optimal methods and better strategies for eliminating mixed pixels are needed. This study demonstrates the potential of using high-resolution UAV RGB images to supplement UAV thermal images for the accurate extraction of maize Tc.

9.
Sensors (Basel) ; 19(13)2019 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-31261734

RESUMO

Rational utilization of water resources is one of the major methods of water conservation. There are significant differences in the irrigation needs of different agricultural fields because of their spatial variability. Therefore, a decision support system for variable rate irrigation (DSS-VRI) by center pivot was developed. This system can process multi-spectral images taken by unmanned aerial vehicles (UAVs) and obtain the vegetation index (VI). The crop evapotranspiration model (ETc) and crop water stress index (CWSI) were obtained from their established relationships with the VIs. The inputs to the fuzzy inference system were constituted with ETc, CWSI and precipitation. To provide guidance for users, the duty-cycle control map was outputted using ambiguity resolution. The control command contained in the map adjusted the duty cycle of the solenoid valve, and then changed the irrigation amount. A water stress experiment was designed to verify the rationality of the DSS-VRI. The results showed that the more severe water stress is, the more irrigation is obtained, consistent with the expected results. Meanwhile, a user-friendly software interface was developed to implement the DSS-VRI function.

10.
J Cell Mol Med ; 23(5): 3505-3511, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30873755

RESUMO

In current study, a bioinformatic-based network pharmacology was used to identify the osteosarcoma (OGS)-pathological targets and formononetin (FN)-treated targets before the main core predictive biotargets were screened. In addition, all core targets were selected through a number of bioinformatic databases, followed by identification of predominant biological processes and signalling pathways of FN anti-OGS. Further, top three core targets of FN anti-OGS were determined as oestrogen receptor 1 (ESR1), tumour protein p53 (TP53), receptor tyrosine-protein kinase erbB-2 (ERBB2) respectively. In clinical biochemical data, the plasma samples of OGS showed the increased trends of alkaline phosphatase, triglyceride, blood glucose, lactate dehydrogenase, high-sensitive C-reactive protein and some immune cell counts when referenced to medical criteria. In clinicopathological examination, histological OGS sections resulted in increased positive cell counts of neoplastic ESR1, TP53, ERBB2. To further validate these corn proteins in experimental study in vivo, FN-treated tumour-bearing nude mice showed intracellular reductions of ESR1, TP53, ERBB2 positive expressions, accompanied with visibly reduced tumour weights. Collectively, our bioinformatic and experimental findings disclosed main core targets, biological processes and signalling pathways of FN anti-OGS. Interestingly, the top core targets were representatively validated following FN treatment in vivo. Therefore, we reasoned that these predictive targets might be the potential biomarkers for screening and treating osteosarcoma.


Assuntos
Antineoplásicos/farmacologia , Biomarcadores Tumorais/antagonistas & inibidores , Neoplasias Ósseas/tratamento farmacológico , Isoflavonas/farmacologia , Terapia de Alvo Molecular/métodos , Osteossarcoma/tratamento farmacológico , Adolescente , Adulto , Fosfatase Alcalina/antagonistas & inibidores , Fosfatase Alcalina/sangue , Animais , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/metabolismo , Neoplasias Ósseas/sangue , Neoplasias Ósseas/metabolismo , Linhagem Celular Tumoral , Biologia Computacional/métodos , Receptor alfa de Estrogênio/antagonistas & inibidores , Receptor alfa de Estrogênio/metabolismo , Feminino , Humanos , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Osteossarcoma/sangue , Osteossarcoma/metabolismo , Receptor ErbB-2/antagonistas & inibidores , Receptor ErbB-2/metabolismo , Carga Tumoral/efeitos dos fármacos , Proteína Supressora de Tumor p53/antagonistas & inibidores , Proteína Supressora de Tumor p53/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
11.
J Cell Physiol ; 234(10): 17305-17313, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30790283

RESUMO

Osteogenic sarcoma (OGS) is a primary bone cancer, characterized by aggressive neoplasm from mesenchymal oncogenesis. However, the clinical therapeutic regimen against OGS is limited. Therefore, potential medication warrants to be further developed. Our previous study indicates that formononetin (FN) exerts effective pharmacological activity against OGS. This study aimed to further decipher the molecular mechanism behind this benefit. Patients with OGS were recruited for clinical data assay and immunoassay. Human OGS cell line (U2OS) and tumor-bearing nude mice were subjected to a battery of biochemical analyses and immunoassays for integrative evaluation of FN-exerted anti-OGS effects. In human data, OGS samples showed increased expressions of ERα, p-PI3KCATyr317 , and p-AKT Ser473 proteins, followed by notably upregulated miR-375 content in comparison with that in OGS-free. In addition, FN-treated U2OS cells showed inhibited cell proliferation, elevated lactic dehydrogenase production and lowered endogenous miR-375 level in cells. Further, reduced immunopositive cells of Ki-67, p-PI3KCA Tyr317 , and p-AKT Ser473 were observed by the treatments of FN, while the intracellular Bax- and Apaf-1-positive cells were increased dose-dependently. Beneficially, FN-treated tumor-bearing mice exhibited reduced tumor mass and intercellular miR-375 expression. Meanwhile, immuno-labeled cells and proteins of Bax, Caspase-3, and Apaf-1 in FN-treated mice were increased dose-dependently, whereas ERα, p-PI3KCA Tyr317 , and p-AKT Ser473 positive cells and proteins were downregulated, respectively. Collectively, our current results elucidate that FN exerts effective therapeutic benefits against OGS, and the pharmacological mechanism may be related to promoting cell apoptosis by inactivating intracellular miR-375/ERα-PI3K/AKT cascaded pathway.


Assuntos
Neoplasias Ósseas/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Isoflavonas/farmacologia , Osteossarcoma/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Neoplasias Ósseas/metabolismo , Linhagem Celular Tumoral , Receptor alfa de Estrogênio/metabolismo , Humanos , Camundongos Nus , Osteossarcoma/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo
12.
Int Orthop ; 40(9): 1883-90, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26987979

RESUMO

PURPOSE: The aim of this study is to compare mTLIF vs. oTLIF with regard to peri-operative complications, operative time, estimated blood loss, fluoroscopic time, and the length of hospital stay. METHODS: The PubMed and EMBASE databases were searched for relevant articles reporting patients undergoing TLIF, and a comparison between mTILF and oTLIF was performed. The database included patient demographic information, complications, operative time, fluoroscopic time, and the length of hospital stay. RESULTS: Fourteen studies were included in this systematic review. The total number of subjects included was 901, of which 455 underwent mTLIF (50 %) and 446 underwent oTLIF (50 %). The operating time for the mTLIF was ranged from 116 to 390 minutes, compared with 102 to 365 minutes for oTLIF, the operating time tended to be longer in the mTLIF group than the oTLIF group. The estimated blood loss was lower in the mTLIF group, ranging from 51 to 578 ml in mTLIF and 225 to 961 ml in oTLIF, respectively. Length of hospital stay was short for the mTLIF with a 2.3 to 10.6 days hospitalization compared to 2.9 to 14.6 days for oTLIF. However the fluoroscopic time was consistently higher in the mTLIF group with a 49 to 106 seconds of fluoroscopy compared to 16.4 to 44 seconds for oTLIF. The complications divided into technical complications and infection complications. The main technical and infection complications included dural tears, screw malposition, and wound infection. Systemic complications included pneumonia, urinary tract infection, and DVT. The numbers of patients with complication was 54 out of 455 (11.87 %) in the mTLIF, and 64 out of 446 (14.35 %) in the oTLIF. CONCLUSION: The review shows mTLIF offers several potential advantages in reducing blood loss and the length of hospital stay, especially lowering the complication rates for patients compared with oTLIF. However, it required much more operative time and radiation exposure. Class I evidence and high-quality randomized controlled trials are needed for further study.


Assuntos
Procedimentos Cirúrgicos Minimamente Invasivos/efeitos adversos , Fusão Vertebral/efeitos adversos , Parafusos Ósseos , Fluoroscopia , Humanos , Tempo de Internação , Vértebras Lombares , Região Lombossacral , Duração da Cirurgia , Estudos Retrospectivos , Fusão Vertebral/métodos , Resultado do Tratamento
13.
Exp Ther Med ; 12(6): 3735-3741, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28105105

RESUMO

Adaptation to hypoxia is an important process physiologically and pathologically. Hypoxia-inducible factor-1α (HIF-1α) participates in the cancer biology of numerous endocrine tumors, including their proliferation and differentiation. In the present study, the hypothesis that HIF-1α promotes tumorigenesis in thyroid cancer via upregulating angiogenesis-associated markers is investigated. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used to examine the expression of HIF-1α in thyroid cancer cell lines, and to detect the expression of WW domain containing E3 ubiquitin protein ligase (WWP)2, WWP9, vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2) in MZ-CRC-1 and TT thyroid cancer cells. Cell proliferation was measured using a Cell Count Kit-8. Cell apoptosis and cell cycle was assessed by flow cytometry. Cell invasive ability was examined by Matrigel transwell analysis. RT-qPCR and western blot analyses demonstrated that the mRNA and protein expression levels of HIF-1α were significant higher in MZ-CRC-1 and TT thyroid cancer cells than in another three thyroid cancer cells (P<0.01). HIF-1α knockdown cells demonstrated inhibition of cell proliferation and invasion, arrested cell cycle at the G1 phase, and induction of cell apoptosis. The protein expression levels of WWP2, WWP9, VEGF and VEGFR2 were decreased in HIF-1α knockdown MZ-CRC-1 and TT cells. In conclusion, HIF-1α may be important in cell apoptosis and invasion of thyroid cancer cells, likely through regulating WWP2, WWP9, VEGF and VEGFR2 expression.

14.
Cell Biochem Biophys ; 71(1): 383-8, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25134663

RESUMO

Akt2 overexpression correlates with chemoresistance of colorectal cancer (CRC). However, the cellular functions and precise signals elicited by Akt2 in LSCC have not been elucidated. Here, we transfected a CRC cell line HCT116 with Akt-2 targeted shRNA in order to establish a cell line with Akt2 knockdown. In vitro experiments showed that knockdown Akt2 in HCT116 cells was associated with decrease in cell proliferation as well as enhanced cell apoptosis. Furthermore, our results demonstrated that Akt2 knockdown correlated with elevated chemosensitivity of HCT116 cells to paclitaxel. Importantly, we found that knockdown of AKt2 resulted in downregulation of MDR-1 and MRP-1. Our findings may lead to a better understanding of the biological effect of Akt2 and may provide mechanistic insights for developing potential therapeutic strategies targeting AKt2.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias Colorretais/patologia , Técnicas de Silenciamento de Genes , Paclitaxel/farmacologia , Proteínas Proto-Oncogênicas c-akt/deficiência , Proteínas Proto-Oncogênicas c-akt/genética , RNA Interferente Pequeno/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Apoptose/genética , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/genética , Células HCT116 , Humanos , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo
15.
Parasitol Int ; 61(1): 187-90, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21777693

RESUMO

Clonorchiasis caused by Clonorchis sinensis is a fish-borne parasitic disease which is endemic in a number of countries. Using the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of C. sinensis as genetic markers, a pair of C. sinensis-specific primers was designed and used to establish a specific PCR assay for the diagnosis of C. sinensis infection in humans, cats and fish. This approach allowed the specific identification of C. sinensis after optimizing amplification conditions, with no amplicons being amplified from related heterogeneous DNA samples, and sequencing of amplicons confirmed the identity of the sequences amplified. The detection limit of this assay was 1.03 pg of adult C. sinensis, 1.1 metacercariae per gram of fish filet, and a single egg in human and cat feces. The PCR assay should provide a useful tool for the diagnosis and molecular epidemiological investigation of clonorchiasis in humans and animals.


Assuntos
Doenças do Gato/parasitologia , Clonorquíase/diagnóstico , Clonorchis sinensis/isolamento & purificação , DNA de Helmintos/isolamento & purificação , Fezes/parasitologia , Doenças dos Peixes/parasitologia , Parasitologia/métodos , Reação em Cadeia da Polimerase/métodos , Animais , Gatos , Clonorchis sinensis/genética , Clonorchis sinensis/crescimento & desenvolvimento , DNA de Helmintos/genética , DNA Espaçador Ribossômico , Peixes , Humanos , Metacercárias/genética , Metacercárias/crescimento & desenvolvimento , Metacercárias/isolamento & purificação , Óvulo/crescimento & desenvolvimento , Sensibilidade e Especificidade
16.
Parasitol Int ; 61(1): 183-6, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21729765

RESUMO

Clonorchiasis caused by the oriental liver fluke Clonorchis sinensis is a fish-borne zoonosis endemic in a number of countries. This article describes the development of a TaqMan based real-time PCR assay for detection of C. sinensis DNA in human feces and in fishes. Primers targeting the first internal transcribed spacer (ITS-1) sequence of the fluke were highly specific for C. sinensis, as evidenced by the negative amplification of closely related trematodes in the test with the exception of Opisthorchis viverrini. The detection limit of the assay was 1pg of purified genomic DNA, 5EPG (eggs per gram feces) or one metacercaria per gram fish filet. The assay was evaluated by testing 22 human fecal samples and 37 fish tissues microscopically determined beforehand, and the PCR results were highly in agreement with the microscopic results. This real-time PCR assay provides a useful tool for the sensitive detection of C. sinensis DNA in human stool and aquatic samples in China and other endemic countries where O. viverrini and Opisthorchis felineus are absent.


Assuntos
Clonorquíase/diagnóstico , Clonorchis sinensis/isolamento & purificação , DNA de Helmintos/isolamento & purificação , Fezes/parasitologia , Parasitologia/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Doenças do Gato/parasitologia , Gatos , China , Clonorchis sinensis/genética , Clonorchis sinensis/crescimento & desenvolvimento , DNA de Helmintos/genética , DNA Espaçador Ribossômico , Doenças dos Peixes/parasitologia , Peixes , Humanos , Óvulo/crescimento & desenvolvimento , Sensibilidade e Especificidade
17.
Parasit Vectors ; 4: 180, 2011 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-21929783

RESUMO

BACKGROUND: Clonorchiasis, caused by Clonorchis sinensis, is one of the major parasitic zoonoses in China, particularly in China's southern Guangdong province where the prevalence of C. sinensis infection in humans is high. However, little is known of the prevalence of C. sinensis infection in its reservoir hosts dogs and cats. Hence, the prevalence of C. sinensis infection in dogs and cats was investigated in Guangdong province, China between October 2006 and March 2008. RESULTS: A total of 503 dogs and 194 cats from 13 administrative regions in Guangdong province were examined by post-mortem examination. The worms were examined, counted, and identified to species according to existing keys and descriptions. The average prevalences of C. sinensis infection in dogs and cats were 20.5% and 41.8%, respectively. The infection intensities in dogs were usually light, but in cats the infection intensities were more serious. The prevalences were higher in some of the cities located in the Pearl River Delta region which is the most important endemic area in Guangdong province, but the prevalences were relatively lower in seaside cities. CONCLUSIONS: The present investigation revealed a high prevalence of C. sinensis infection in its reservoir hosts dogs and cats in China's subtropical Guangdong province, which provides relevant "base-line" data for conducting control strategies and measures against clonorchiasis in this region.


Assuntos
Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Clonorquíase/veterinária , Clonorchis sinensis/isolamento & purificação , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Animais , Gatos , China/epidemiologia , Clonorquíase/epidemiologia , Clonorquíase/parasitologia , Cães , Prevalência
18.
Rapid Commun Mass Spectrom ; 22(6): 766-72, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18275095

RESUMO

Terpene lactones are a family of compounds with unique chemical structures, first recognised in an extract of Ginkgo biloba. The discovery of terpene lactone derivatives has recently been reported in more and more plant extracts and even food products. In this study, mass spectrometric characteristics of the standard terpene lactones in Ginkgo biloba were comprehensively studied using both an ion trap and a quadrupole time-of-flight (QTOF) mass spectrometer. The mass spectral fragmentation data from both techniques was compared to obtain the mass spectrometric fragmentation pathways of the terpene lactones with high confidence. The data obtained will facilitate the analysis and identification of terpene lactones in future plant research via the fragmentation knowledge reported here.


Assuntos
Ginkgo biloba/química , Lactonas/química , Extratos Vegetais/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Terpenos/química
19.
Rapid Commun Mass Spectrom ; 20(24): 3619-24, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17094168

RESUMO

Ginkgo biloba is one of the most popular herbal nutritional supplements, with terpene lactones and flavonoids being the two major active components. An on-line purification high-performance liquid chromatography/mass spectrometry (HPLC/MS) method was successfully developed for the quantitative determination of flavonoids and terpene lactones excreted in human urine after ingesting the herbal supplement. Satisfactory separation was obtained using a C18 capillary column made in-house with sample clean-up and pre-concentration achieved using a C18 pre-column with column switching. High selectivity and limits of detection of 1-18 ng/mL were achieved using a selected ion monitoring (SIM) scan in negative ion mode; the on-line solid-phase extraction (SPE) recovery of the active components in Ginkgo biloba determined in this study was greater than 75%.


Assuntos
Eletrocromatografia Capilar/métodos , Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/urina , Ginkgo biloba/química , Espectrometria de Massas/métodos , Extratos Vegetais/urina , Urinálise/métodos , Eletrocromatografia Capilar/instrumentação , Cromatografia Líquida de Alta Pressão/instrumentação , Humanos , Espectrometria de Massas/instrumentação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Urinálise/instrumentação
20.
Rapid Commun Mass Spectrom ; 20(18): 2753-60, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16921563

RESUMO

A reversed-phase high-performance liquid chromatography/electrospray ionisation mass spectrometry (RP-HPLC/ESI-MS) method was developed and validated for the simultaneous determination of ten major active components in Ginkgo biloba extract (bilobalide, ginkgolides A, B, C, quercetin, kaempferol, isorhamnetin, rutin hydrate, quercetin-3-beta-D-glucoside and quercitrin hydrate) which have not been previously reported to be quantified in a single analysis. The ten components exhibit baseline separation in 50 min by C18 chromatography using a water/1:1 (v/v) methanol/acetonitrile gradient. Quantitation was performed using negative ESI-MS in selected ion monitoring (SIM) mode. Good reproducibility and recovery were obtained by this method. The sensitivity of both UV and different mass spectrometry modes (full scan, selected ion monitoring (SIM), and selected reaction monitoring (SRM)) were compared and both quantitation with and without internal standard were evaluated. The analysis of Ginkgo biloba commercial products showed remarkable variations in the rutin and quercetin content as well as the terpene lactone contents although all the products satisfy the conventional quality control method.


Assuntos
Cromatografia Líquida de Alta Pressão , Suplementos Nutricionais/análise , Ginkgo biloba , Extratos Vegetais/análise , Espectrometria de Massas por Ionização por Electrospray/métodos , Ciclopentanos/análise , Flavonóis/análise , Furanos/análise , Ginkgolídeos/análise , Quempferóis/análise , Quercetina/análogos & derivados , Quercetina/análise , Rutina/análise
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