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1.
Molecules ; 26(4)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33572420

RESUMO

Breast cancer is one of the major cancers of women in the world. Despite significant progress in its treatment, an early diagnosis can effectively reduce its incidence rate and mortality. To improve the reliability of Raman-based tumor detection and analysis methods, we conducted an ex vivo study to unveil the compositional features of healthy control (HC), solid papillary carcinoma (SPC), mucinous carcinoma (MC), ductal carcinoma in situ (DCIS), and invasive ductal carcinoma (IDC) tissue samples. Following the identification of biological variations occurring as a result of cancer invasion, principal component analysis followed by linear discriminate analysis (PCA-LDA) algorithm were adopted to distinguish spectral variations among different breast tissue groups. The achieved results confirmed that after training, the constructed classification model combined with the leave-one-out cross-validation (LOOCV) method was able to distinguish the different breast tissue types with 100% overall accuracy. The present study demonstrates that Raman spectroscopy combined with multivariate analysis technology has considerable potential for improving the efficiency and performance of breast cancer diagnosis.


Assuntos
Adenocarcinoma Mucinoso/patologia , Neoplasias da Mama/classificação , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/patologia , Carcinoma Intraductal não Infiltrante/patologia , Carcinoma Papilar/patologia , Análise Espectral Raman/métodos , Adenocarcinoma Mucinoso/cirurgia , Algoritmos , Neoplasias da Mama/cirurgia , Carcinoma Ductal de Mama/cirurgia , Carcinoma Intraductal não Infiltrante/cirurgia , Carcinoma Papilar/cirurgia , Estudos de Casos e Controles , Análise Discriminante , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Análise de Componente Principal
2.
J Physiol ; 593(16): 3739-55, 2015 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-26110238

RESUMO

KEY POINTS: Neuropathic pain resulting from peripheral nerve injury is initiated and maintained by persistent ectopic activity in primary afferent neurons. Sciatic nerve injury increases the excitability of medium-sized dorsal root ganglion (DRG) neurons. Levels of the inflammatory cytokine interleukin 1ß (IL-1ß) increase and peak after 7 days. Five to six days of exposure of medium sized DRG neurons to 100 pm IL-1ß promotes persistent increases in excitability which abate within 3-4 days of cytokine removal. This is associated with a profound attenuation of K(+) channel currents but only modest increases in function of cyclic nucleotide-sensitive hyperpolarization-activated channels (HCNs) and of voltage-gated Na(+) and Ca(2+) channel currents. It is unlikely, therefore, that direct interaction of IL-1ß with DRG neurons is capable of initiating an enduring phenotypic shift in their electrophysiological properties that follows sciatic nerve injury. The findings also underline the importance of K(+) channel modulation in the actions of inflammatory mediators on peripheral neurons. ABSTRACT: Chronic constriction injury of rat sciatic nerve promotes signs of neuropathic pain. This is associated with an increase in the level of interleukin 1ß (IL-1ß) in primary afferents that peaks at 7 days. This initial cytokine exposure has been proposed to trigger an enduring alteration in neuronal phenotype that underlies chronic hyper-excitability in sensory nerves, which initiates and maintains chronic neuropathic pain. We have shown previously that 5-6 days of exposure of rat dorsal root ganglia (DRGs) to 100 pm IL-1ß increases the excitability of medium-sized neurons. We have now found using whole-cell recording that this increased excitability reverts to control levels within 3-4 days of cytokine removal. The effects of IL-1ß were dominated by changes in K(+) currents. Thus, the amplitudes of A-current, delayed rectifier and Ca(2+) -sensitive K(+) currents were reduced by ∼68%, ∼64% and ∼36%, respectively. Effects of IL-1ß on other cation currents were modest by comparison. There was thus a slight decrease in availability of high voltage-activated Ca(2+) channel current, a small increase in rates of activation of hyperpolarization-activated cyclic nucleotide-gated channel current (IH ), and a shift in the voltage dependence of activation of tetrodotoxin-sensitive sodium current (TTX-S INa ) to more negative potentials. It is unlikely, therefore, that direct interaction of IL-1ß with DRG neurons initiates an enduring phenotypic shift in their electrophysiological properties following sciatic nerve injury. Persistent increases in primary afferent excitability following nerve injury may instead depend on altered K(+) channel function and on the continued presence of slightly elevated levels IL-1ß and other cytokines.


Assuntos
Gânglios Espinais/fisiologia , Interleucina-1beta/farmacologia , Neurônios/efeitos dos fármacos , Canais de Potássio/fisiologia , Animais , Células Cultivadas , Masculino , Neurônios/fisiologia , Ratos Sprague-Dawley
3.
Zhong Yao Cai ; 36(8): 1340-3, 2013 Aug.
Artigo em Zh | MEDLINE | ID: mdl-24558834

RESUMO

OBJECTIVE: To investigate the optimal technology of extraction of fufang zhenqin feining capsules and establish a method for determination its oleanolic acid. METHODS: Orthogonal test was employed for selecting the optimum extraction technology by the index of the content of oleanolic acid in extraction. HPLC was performed on a C18 (250 mm x 4.6 mm, 5 microm) column using methanol-0.3% triethylamine solution (80: 20) as the mobile phase at a flow rate of 1 ml/min. The detection wavelength was set at 210 nm. The column temperature was 30 degrees C and the injection size was 25 microL. RESULTS: The optimum extraction technology was as follows: the concertration of alcohol was 70%, the ratio of solid/material was 8: 1, extracting 3 times with half an hour for each time. The linear range of oleanolic acid was 59.68 - 596.8 microg/ml (r = 0.9997), the average recovery was 99.79% (RSD = 0.5%). CONCLUSION: The considerable extraction rate of active components in the drugs is achieved by applying the selected technology, and the simple method is fit for production. The established HPLC method is simple, accurate, special and can be used for the quality control of fufang zhenqin feining capsules.


Assuntos
Ácido Oleanólico/análise , Cápsulas , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Controle de Qualidade
4.
Expert Opin Drug Saf ; 22(8): 707-714, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36803342

RESUMO

BACKGROUND: Romiplostim and eltrombopag are thrombopoietin receptor agonists (TPORAs) that have been approved by the FDA on 22 August 2008 and 20 November 2008 for pediatric immune thrombocytopenia (ITP). However, postmarketing pharmacovigilance of TPORAs in children still attracts much attention. We aimed to evaluate the safety of the TPORAs romiplostim and eltrombopag using data from the Adverse Event Reporting System database of FDA (FAERS). RESEARCH DESIGN AND METHODS: We conducted a disproportionality analysis and analyzed data from the FAERS database to characterize the key features of adverse events (AEs) associated with TPO-RAs approved for children under 18 years of age. RESULTS: Since their approval in the market in 2008, 250 and 298 reports of romiplostim and eltrombopag use in children have been published in the FAERS database, respectively. The most frequent AE associated with romiplostim and eltrombopag was epistaxis. Neutralizing antibodies and vitreous opacities showed the strongest signals for romiplostim and eltrombopag, respectively. CONCLUSIONS: The labeled AEs for romiplostim and eltrombopag in children were analyzed. Unlabeled AEs may reflect the potential of new clinical individuals. Early recognition and management of AEs that appear in children treated with romiplostim and eltrombopag are of key importance in clinical practice.


Assuntos
Púrpura Trombocitopênica Idiopática , Trombocitopenia , Humanos , Criança , Adolescente , Púrpura Trombocitopênica Idiopática/tratamento farmacológico , Púrpura Trombocitopênica Idiopática/induzido quimicamente , Farmacovigilância , Receptores de Trombopoetina/agonistas , Receptores de Trombopoetina/uso terapêutico , Trombocitopenia/induzido quimicamente , Benzoatos/efeitos adversos , Proteínas Recombinantes de Fusão/efeitos adversos
5.
Zhonghua Gan Zang Bing Za Zhi ; 20(3): 193-5, 2012 Mar.
Artigo em Zh | MEDLINE | ID: mdl-22475138

RESUMO

OBJECTIVE: To investigate the clinical characteristics and responsible agents of drug-induced liver injury (DILI) in pediatric patients. METHODS: Thirty-one cases of DILI treated in our hospital's pediatric ward were retrospectively analyzed. The clinical data for each patient were extracted from the patient's medical records, and included reported causes, physical and biochemical features, natural history, blood examination results, and hepatic pathology findings. RESULTS: The 31 pediatric cases of DILI accounted for 1.7% of the 1831 total cases of drug-induced liver injury treated at our hospital between February 2002 to June 2011. The pediatric DILI population was composed of 20 males and 11 females, with an average age of 8.8+/-3.9 years old (range, 0.3-14.0). The liver injury patterns represented among the cases were: hepatocellular (25.8%), cholestasis (25.8%), and mixed hepatocellular-cholestatic (48.4%). Antimicrobials were the most common cause (41.9%) of DILI, followed by the herbal medicine (29.0%) and febrifuge drugs (19.4%). A single drug was implicated in nine cases (29.0%), and two or more drugs were implicated in 22 cases (71%). Most of the children had good prognosis, but those with pre-existing disease had poor prognosis. One child died of hepatic failure, making the death rate 3.23%. The average hospitalization time was 25.2 days, and the patients with hepatocellular injury had shorter hospitalization time than those with mixed injury. CONCLUSION: Drug-induced liver injury in our pediatric population was most often caused by antimicrobials, followed by herbal medicine and febrifuge drugs. Most patients presented with mixed hepatocellular-cholestatic injury. Children with pre-existing diseases or hepatic failure had poor prognosis.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/diagnóstico , Doença Hepática Induzida por Substâncias e Drogas/patologia , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Prognóstico , Estudos Retrospectivos
6.
J Photochem Photobiol B ; 226: 112366, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34826719

RESUMO

Confocal Raman Microspectroscopy (CRM) was employed to clarify the cellular response of cisplatin in osteosarcoma (OS) cells with different dosages and incubation times. The K7M2 mouse osteosarcoma cells were treated by cisplatin in 0 µM (UT group), 20 µM (20 T group), and 40 µM (40 T group) doses for 24-h (24H group) and 48-h (48H group), respectively. Raman spectroscopy was utilized to analyze the drug induced variations of intracellular biochemical components in osteosarcoma cells. The spectral results shows that the main changes in its biochemical composition come from nucleic acids. By adopting three different kernel functions (linear, polynomial, and Gaussian radial basis function (RBF)), principal component analysis combined with support vector machine models (PCA-SVM) was built to address the spectral variations among all investigated groups. Meanwhile, multivariate curve resolution alternating least squares (MCR-ALS) was further utilized to discuss on the chemical interpretation on the acquired spectral results. Moreover, Raman spectral images, which is reconstructed by K-means cluster analysis (KCA) with point-scanned hyperspectral dataset, was applied to illustrate the drug induced compositional and morphological variations in each subcellular region. The achieved results not only prove the application potential of Raman based analytical technique in non-labeled intracellular studies, but also illustrate the detailed compositional and structural information of cisplatin induced OS cell responses from the perspective of multivariate analysis and imaging of Raman spectroscopy.


Assuntos
Cisplatino
7.
J Biophotonics ; 15(12): e202200189, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36057844

RESUMO

By using Raman microspectroscopy, it aims to elucidate the cellular variations caused by the combination drug of γ-secretase inhibitor (DAPT) and cisplatin in osteosarcoma (OS) cells. Illustrated by the obtained results of spectral analysis, the intracellular composition significantly changed after combined drug actions compared to the solo DAPT treatment, indicating the synergistic effect of DAPT combined with cisplatin on OS cells. Meanwhile, multivariate curve resolution-alternating least squares (MCR-ALS) algorithm was utilized to address the biochemical constitution changes in all investigated groups including the untreated (UT), DAPT (40D) and combined drug (40D + 20C) treated cells. K-means cluster and univariate imaging were both utilized to visualize the changes in subcellular morphology and biochemical distribution. The presented study provides a unique understanding on the cellular responses to DAPT combined with cisplatin from the natural biochemical perspectives, and laids an experimental foundation for exploring the therapeutic strategies of other combined anticancer drugs in cancer cell model.


Assuntos
Neoplasias Ósseas , Osteossarcoma , Humanos , Secretases da Proteína Precursora do Amiloide , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/patologia , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Osteossarcoma/tratamento farmacológico , Osteossarcoma/patologia , Inibidores da Agregação Plaquetária/uso terapêutico , Antinematódeos/uso terapêutico
8.
Materials (Basel) ; 15(12)2022 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-35744191

RESUMO

The present study is intended to develop and test a cost-effective and efficient printing method for fabricating flexible metamaterial film with high electromagnetic wave absorptivity. The film can be easily applied to the surfaces with curved aspects. Firstly, numerical parametric study of the absorption characteristics of the film is performed for the range of frequency varying from 2.0 to 9.0 GHz based on commercial software package. Secondly, the flexible metamaterial films are fabricated, and experiments are conducted. The flexible metamaterial film consists of a flexible dielectric film made of polyimide (PI) and an array of split-ring resonators. The split-ring resonators of different geometric dimensions are fabricated on the PI film surface by using a silver nanoparticles ink jet printer. The performance of the flexible structure is then measured and dependence of operation frequency with higher absorptivity on the dimensions of the split-ring resonators is investigated. A comparison between the numerical and experimental data shows that the numerical predictions of the operation frequency with higher absorptivity closely agree with the experimental data.

9.
mSystems ; 7(3): e0020922, 2022 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-35615956

RESUMO

The rapid emergence of multidrug-resistant/extensively drug-resistant tuberculosis (TB) is responsible for treatment failure in patients with TB and significantly endangers global public health. Recently, bioenergetics has become a new paradigm for anti-TB drug discovery and is based on the link between bacterial ATP levels and drug efficacy. A better understanding of the role of ATP fluctuations during antibiotic treatment may provide insight into antibiotic-mediated killing of mycobacteria. Here, we employed an advanced single-fluorescence FRET (fluorescence resonance energy transfer)-based ATP biosensor, ATPser, for the stable and convenient detection of intracellular ATP fluctuations in mycobacteria. This strategy correlated closely with the results obtained from conventional luminescence ATP assays, indicating the reliability of the system for bioenergetics analysis in mycobacteria. Moreover, the reporter strains expressing ATPser displayed obvious ATP changes when subjected to different stresses, such as starvation and ATP depletion. Interestingly, we observed that different antibiotics induced fluctuations in cellular ATP levels in individual cells of various magnitudes, revealing a strong connection between ATP fluctuations and drug efficacy. Furthermore, drug combinations accelerated ATP perturbation, resulting in increased cell death. We concluded that ATPser enabled real-time measurement of ATP at the single-cell level in mycobacteria, and monitoring ATP dynamics in drug-treated bacteria may shed light on novel treatment strategies. IMPORTANCE Bioenergetics has emerged as a new paradigm for antituberculosis (anti-TB) drug discovery, and the cellular ATP level is the core indicator reflecting bacterial metabolic homeostasis. Although several bulk assays have been designed for the measurement of cellular ATP content, a more convenient strategy is required for real-time ATP measurement of single viable cells. In this study, by combining the ε-subunit of Bacillus subtilis FoF1-ATP synthase with a circularly permuted green fluorescent protein [(cp)GFP], we constructed a FRET-based single-fluorescence ATP sensor, ATPser, for real-time single-cell ATP detection among a mycobacterial population. Using the ATPser, we designed different drug combinations containing components that have similar/opposite effects on ATP alternation. Our results demonstrated that increased cellular ATP fluctuations were associated with depletion of mycobacterial viability, while counteracting ATP fluctuations weakened the killing effect of the drug regime. Thus, potentially efficient drug combinations can be considered based on their similar effects on mycobacterial ATP levels, and ATPser may be a useful tool to study mycobacterial bioenergetics and to guide drug regime design.


Assuntos
Transferência Ressonante de Energia de Fluorescência , Mycobacterium , Humanos , Reprodutibilidade dos Testes , Mycobacterium/metabolismo , Antituberculosos/farmacologia , Trifosfato de Adenosina/metabolismo
10.
Microbiol Spectr ; 10(1): e0132221, 2022 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-35019676

RESUMO

Clostridioides difficile, which causes life-threatening diarrheal disease, presents an urgent threat to health care systems. In this study, we present a retrospective genomic and epidemiological analysis of C. difficile in a large teaching hospital. First, we collected 894 nonduplicate fecal samples from patients during a whole year to elucidate the C. difficile molecular epidemiology. We then presented a detailed description of the population structure of C. difficile based on 270 isolates separated between 2015 and 2020 and clarified the genetic and phenotypic features by MIC and whole-genome sequencing. We observed a high carriage rate (19.4%, 173/894) of C. difficile among patients in this hospital. The population structure of C. difficile was diverse with a total of 36 distinct STs assigned. In total, 64.8% (175/270) of the isolates were toxigenic, including four CDT-positive (C. difficile transferase) isolates, and 50.4% (135/268) of the isolates were multidrug-resistant. Statistically, the rates of resistance to erythromycin, moxifloxacin, and rifaximin were higher for nontoxigenic isolates. Although no vancomycin-resistant isolates were detected, the MIC for vancomycin was higher for toxigenic isolates (P < 0.01). The in-hospital transmission was observed, with 43.8% (110/251) of isolates being genetically linked to a prior case. However, no strong correlation was detected between the genetic linkage and epidemiological linkage. Asymptomatic colonized patients play the same role in nosocomial transmission as infected patients, raising the issue of routine screening of C. difficile on admission. This work provides an in-depth description of C. difficile in a hospital setting and paves the way for better surveillance and effective prevention of related diseases in China. IMPORTANCE Clostridioides difficile infections (CDI) are the leading cause of healthcare-associated diarrhea and are known to be resistant to multiple antibiotics. In the past decade, C. difficile has emerged rapidly and has spread globally, causing great concern among American and European countries. However, research on CDI remains limited in China. Here, we characterized the comprehensive spectrum of C. difficile by whole-genome sequencing (WGS) in a Chinese hospital, showing a high detection rate among patients, diverse genome characteristics, a high level of antibiotic resistance, and an unknown nosocomial transmission risk of C. difficile. During the study period, two C. difficile transferase (CDT)-positive isolates belonging to a new multilocus sequence type (ST820) were detected, which have caused serious clinical symptoms. This work describes C. difficile integrally and provides new insight into C. difficile surveillance based on WGS in China.


Assuntos
Antibacterianos/farmacologia , Clostridioides difficile/efeitos dos fármacos , Clostridioides difficile/genética , Infecções por Clostridium/microbiologia , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana , Adolescente , Adulto , Idoso , Proteínas de Bactérias/genética , Criança , Pré-Escolar , China/epidemiologia , Clostridioides difficile/classificação , Clostridioides difficile/isolamento & purificação , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/transmissão , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/transmissão , Eritromicina/farmacologia , Feminino , Genoma Bacteriano , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Moxifloxacina/farmacologia , Filogenia , Estudos Retrospectivos , Rifaximina/farmacologia , Vancomicina/farmacologia , Sequenciamento Completo do Genoma , Adulto Jovem
11.
J Neurosci Res ; 89(5): 689-99, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21337369

RESUMO

A significant loss of neurons in the dorsal root ganglia (DRG) has been reported in animal models of peripheral nerve injury. Neonatal sensory neurons are more susceptible than adult neurons to axotomy- or nerve growth factor (NGF) withdrawal-induced cell death. To develop therapies for preventing irreversible sensory cell loss, it is essential to understand the molecular mechanisms responsible for DRG cell death and survival. Here we describe how the expression of the growth arrest- and DNA damage-inducible gene 45α (GADD45A) is correlated with neuronal survival after axotomy in vivo and after NGF withdrawal in vitro. GADD45A expression is low at birth and does not change significantly after spinal nerve ligation (SNL). In contrast, GADD45A is robustly up-regulated in the adult rat DRG 24 hr after SNL, and this up-regulation persists as long as the injured fibers are prevented from regenerating. In vitro delivery of GADD45A protects neonatal rat DRG neurons from NGF withdrawal-induced cytochrome c release and cell death. In addition, in vivo knockdown of GADD45A expression in adult injured DRG by small hairpin RNA increased cell death. Our results indicate that GADD45A protects neuronal cells from SNL-induced cell death.


Assuntos
Proteínas de Ciclo Celular/metabolismo , Gânglios Espinais/metabolismo , Degeneração Neural/patologia , Degeneração Neural/prevenção & controle , Proteínas Nucleares/metabolismo , Doenças do Sistema Nervoso Periférico/patologia , Doenças do Sistema Nervoso Periférico/prevenção & controle , Células Receptoras Sensoriais/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Axotomia/métodos , Proteínas de Ciclo Celular/genética , Morte Celular/genética , Modelos Animais de Doenças , Gânglios Espinais/patologia , Ligantes , Masculino , Degeneração Neural/metabolismo , Fator de Crescimento Neural/deficiência , Proteínas Nucleares/genética , Doenças do Sistema Nervoso Periférico/metabolismo , Ratos , Ratos Sprague-Dawley , Neuropatia Ciática/genética , Neuropatia Ciática/metabolismo , Neuropatia Ciática/prevenção & controle , Células Receptoras Sensoriais/metabolismo , Regulação para Cima/genética
12.
J Biophotonics ; 14(5): e202000456, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33547854

RESUMO

It is a practical necessity for non-professional users to interpret biologically derived Raman spectral information for obtaining accurate and reliable analytical results. An integrated Raman spectral analysis software (NWUSA) was developed for spectral processing, analysis, and feature recognition. It provides a user-friendly graphical interface to perform the following preprocessing tasks: spectral range selection, cosmic ray removal, polynomial fitting based background subtraction, Savitzky-Golay smoothing, area-under-curve normalization, mean-centered procedure, as well as multivariate analysis algorithms including principal component analysis (PCA), linear discriminant analysis, partial least squares-discriminant analysis, support vector machine (SVM), and PCA-SVM. A spectral dataset obtained from two different samples was utilized to evaluate the performance of the developed software, which demonstrated that the analysis software can quickly and accurately achieve functional requirements in spectral data processing and feature recognition. Besides, the open-source software can not only be customized with more novel functional modules to suit the specific needs, but also benefit many Raman based investigations, especially for clinical usages.


Assuntos
Análise Espectral Raman , Máquina de Vetores de Suporte , Algoritmos , Análise Discriminante , Análise dos Mínimos Quadrados , Análise de Componente Principal
13.
Anal Methods ; 13(22): 2527-2536, 2021 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-34008598

RESUMO

Confocal Raman microspectral analysis and imaging were used to elucidate the drug response of osteosarcoma (OS) to cisplatin. Raman spectral data were obtained from OS cells that were untreated (UT group) and treated with 20 µM (20T group) and 40 µM (40T group) cisplatin for 24 hours. Statistical analysis of the changes in specific Raman signals was performed using a one-way ANOVA and multiple Tukey's honest significant difference (HSD) post hoc tests. Principal component analysis-linear discriminant analysis (PCA-LDA) was used to highlight the featured cellular drug responses based on the obtained spectral information. For spectral imaging analysis, k-means cluster analysis (KCA) was adopted to clarify the effect of cisplatin dose changes on the subcellular structure and its biochemical composition. The results suggest that the major biochemical changes induced by cisplatin in OS cells undergoing apoptosis are reduced protein and nucleic acid content. Through univariate analysis, the changes in the distribution of nucleic acids in OS cells induced by different doses of cisplatin were obtained. The combination of Raman spectroscopy and multivariate analysis shows that cisplatin mainly acts on the nucleus and causes changes in the secondary structure of proteins. These results indicate that Raman imaging technology has the potential to offer the basis of dose optimization for personalized cancer treatment by helping to understand in vitro cellular drug interactions.


Assuntos
Neoplasias Ósseas , Osteossarcoma , Preparações Farmacêuticas , Neoplasias Ósseas/diagnóstico por imagem , Cisplatino/farmacologia , Humanos , Osteossarcoma/diagnóstico por imagem , Análise de Componente Principal
14.
J Biomed Sci ; 17: 44, 2010 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-20525259

RESUMO

BACKGROUND: Gene transfer into many cell types has been successfully used to develop alternative and adjunct approaches to conventional medical treatment. However, effective transfection of postmitotic neurons remains a challenge. The aim of this study was to develop a method for gene transfer into rat primary dorsal root ganglion neurons using sonoporation. METHODS: Dissociated cells from adult rat dorsal root ganglion (DRG) cells were sonicated for 1-8 s at 2.5-10 W to determine the optimal ultrasound duration and power for gene transfection and cell survival. Transfection efficiency was compared between sonoporation, liposome and lentiviral vector gene transfer techniques. RESULTS: The optimum ultrasound intensity was 5 W for 2 s and yielded an efficiency of gene transfection of 31% and a survival rate of 35%. CONCLUSIONS: Sonoporation can be optimized to minimize cell death and yield a high percentage of transfected neurons and that this technique can be easily applied to primary cultures of rat dorsal root ganglion neurons.


Assuntos
Gânglios Espinais/citologia , Gânglios Espinais/metabolismo , Técnicas de Transferência de Genes , Animais , Sobrevivência Celular , Células Cultivadas , Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Lentivirus/genética , Lipossomos , Neurônios/citologia , Neurônios/metabolismo , Ratos , Proteínas Recombinantes/genética , Transfecção , Tubulina (Proteína)/metabolismo , Ultrassom
15.
Biomed Opt Express ; 11(2): 1061-1072, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32133237

RESUMO

Lung cancer is the leading cause of death in cancer patients, and microwave ablation (MWA) has been extensively used in clinical treatment. In this study, we characterized the spectra of MWA-treated and untreated lung squamous cell carcinoma (LSCC) tissues, as well as healthy lung tissue, and conducted a preliminary analysis of spectral variations associated with MWA treatment. The results of characteristic spectral analysis of different types of tissues indicated that MWA treatment induces an increase in the content of nucleic acids, proteins, and lipid components in lung cancer tissues. The discriminant model based on the principal component analysis - linear discriminant analysis (PCA-LDA) algorithm together with leave-one-out cross validation (LOOCV) method yield the sensitivities of 90%, 80%, and 96%, and specificities of 86.2%, 93.8%, and 100% among untreated and MWA-treated cancerous tissue, and healthy lung tissue, respectively. These results indicate that Raman spectroscopy combined with multivariate analysis techniques can be used to explore the biochemical response mechanism of cancerous tissue to MWA therapy.

16.
J Neurophysiol ; 102(6): 3203-15, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19793881

RESUMO

Injury or section of a peripheral nerve can promote chronic neuropathic pain. This is initiated by the appearance and persistence of ectopic spontaneous activity in primary afferent neurons that promotes a secondary, enduring increase in excitability of sensory circuits in the spinal dorsal horn ("central sensitization"). We have previously shown that 10-20 days of chronic constriction injury (CCI) of rat sciatic nerve produce a characteristic "electrophysiological signature" or pattern of changes in synaptic excitation of five different electrophysiologically defined neuronal phenotypes in the substantia gelatinosa of the dorsal horn. Although axotomy and CCI send different signals to the dorsal horn, we now find, using whole cell recording, that the "electrophysiological signature" produced 12-22 days after sciatic axotomy is quite similar to that seen with CCI. Axotomy thus has little effect on resting membrane potential, rheobase, current-voltage characteristics, or excitability of most neuron types; however, it does decrease excitatory synaptic drive to tonic firing neurons, while increasing that to delay firing neurons. Since many tonic neurons are GABAergic, whereas delay neurons do not contain gamma-aminobutyric acid, axotomy may reduce synaptic excitation of inhibitory neurons while increasing that of excitatory neurons. Further analysis of spontaneous and miniature (tetrodotoxin-resistant) excitatory postsynaptic currents is consistent with the possibility that decreased excitation of tonic neurons reflects loss of presynaptic contacts. By contrast, increased excitation of "delay" neurons may reflect increased frequency of discharge of presynaptic action potentials. This would explain how synaptic excitation of tonic cells decreases despite the fact that axotomy increases spontaneous activity in primary afferent neurons.


Assuntos
Axotomia/métodos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Nervo Isquiático/fisiologia , Substância Gelatinosa/fisiologia , Sinapses/fisiologia , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Animais Recém-Nascidos , Biofísica , Proteínas de Ligação ao Cálcio/metabolismo , Estimulação Elétrica/métodos , Antagonistas de Aminoácidos Excitatórios/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Antagonistas GABAérgicos/farmacologia , Potenciais Pós-Sinápticos Inibidores/efeitos dos fármacos , Laminectomia/métodos , Proteínas dos Microfilamentos , Técnicas de Patch-Clamp/métodos , Piridazinas/farmacologia , Ratos , Sinapses/efeitos dos fármacos
17.
Biosens Bioelectron ; 24(1): 136-40, 2008 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-18490150

RESUMO

A piezoelectric immunosensor assay was developed with immobilizing immunoglobulin G (IgG) as a probe to detect Schistosoma japonicum circulating antigens (SjCAg). Analytical strategy utilizes the polyclonal antibodies with broad-spectrum recognition to a complex target with high specificity. The immobilized antibodies were purified from immunized rabbit's sera (im-S) and infected rabbit's sera (inf-S) by S. japonicum. The detection capacities of antibodies were compared between the sera of different phenotypes and purified fractions. The sample dilution ratios were also evaluated and optimized. Additionally, the sera with a variety of infection degrees for validation could be discriminated quantitatively. The linear dose-response relationship indicates that the systematic sensitivity of this method is below 150 Hz and the lowest limit of detectable range is above 500 cercariae of S. japonicum infection for 2 weeks. The novel immunosensor technique is well potential to determine the SjCAg in serum samples for clinical diagnosis of parasitosis in early stage.


Assuntos
Antígenos de Helmintos/sangue , Técnicas Biossensoriais/métodos , Imunoensaio/métodos , Schistosoma japonicum/imunologia , Animais , Humanos , Quartzo/química
18.
Anesth Analg ; 102(6): 1765-7, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16717323

RESUMO

We evaluated paravertebral transcutaneous electrical nerve stimulation (TENS) as a means of enhancing anesthesia during hysterectomy. Patients were randomly assigned to experimental (n = 21) and control (n = 20) groups. Anesthesia with isoflurane was performed uniformly for all patients. Paravertebral (T6 and T7) TENS (50 mA, 15 Hz, continuously) was applied in the experimental group. After 15 min of isoflurane, a lower abdominal, skin-to-adipose-tissue incision was made. Seventeen of 21 patients in the experimental group showed no arm or leg movements during the incision, compared to 8 with 20 patients in the control group (P = 0.007). TENS deserves further exploration as an adjunct technique for general anesthesia.


Assuntos
Anestesia Geral , Anestésicos Inalatórios , Extremidades , Isoflurano , Movimento , Estimulação Elétrica Nervosa Transcutânea , Adulto , Feminino , Humanos , Histerectomia , Pessoa de Meia-Idade
19.
Exp Neurol ; 279: 168-177, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26948545

RESUMO

Synapses transmitting nociceptive information in the spinal dorsal horn undergo enduring changes following peripheral nerve injury. Indeed, such injury alters the expression of the GluA2 subunit of glutamatergic AMPA receptors (AMPARs) in the substantia gelatinosa and this predicts altered channel conductance and calcium permeability, leading to an altered function of excitatory synapses. We therefore investigated the functional properties of synaptic AMPA receptors in rat substantia gelatinosa neurons following 10-20d chronic constriction injury (CCI) of the sciatic nerve; a model of neuropathic pain. We measured their single-channel conductance and sensitivity to a blocker of calcium permeable AMPA receptors (CP-AMPARs), IEM1460 (50µM). In putative inhibitory, tonic firing neurons, CCI reduced the average single-channel conductance of synaptic AMPAR from 14.4±3.5pS (n=12) to 9.2±1.0pS (n=10, p<0.05). IEM1460 also more effectively antagonized evoked, spontaneous and miniature EPSCs in tonic neurons from sham operated animals than in those from animals that had been subjected to CCI. By contrast, CCI did not change the effectiveness of IEM1460 in delay firing neurons although average single channel conductance was increased from 7.6±1.2pS (n=11) to 12.2±1.5pS (n=10, p<0.01). CCI thus elicits plastic changes in a specific set of glutamatergic synapses of substantia gelatinosa due to subunit recomposition and loss of GluA2-lacking CP-AMPAR. These insights reveal a molecular mechanism of nerve injury acting at synapses of inhibitory neurons to reduce their drive and therefore inhibitory tone in the spinal cord, therefore contributing to the central sensitization associated with neuropathic pain.


Assuntos
Cálcio/metabolismo , Constrição Patológica/metabolismo , Neuralgia/metabolismo , Neurônios/metabolismo , Receptores de AMPA/metabolismo , Substância Gelatinosa/metabolismo , Sinapses/metabolismo , Adamantano/análogos & derivados , Adamantano/farmacologia , Animais , Antagonistas de Aminoácidos Excitatórios/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Masculino , Ratos , Receptores de AMPA/antagonistas & inibidores , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , Sinapses/efeitos dos fármacos
20.
Chin J Integr Med ; 22(9): 696-703, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27614452

RESUMO

OBJECTIVE: To study a non-drug therapy for hypertension disease by combining percutaneous laser and electric pulse stimulation to acupoint with music, and to test the efficiency of the combining treatment to grade 1 essential hypertension. METHODS: A total of 174 patients with grade 1 essential hypertension were randomly assigned to 3 groups with a random number table after Chinese medicine (CM) syndrome differentiation: the photoelectric and musical treatment group (Group 1, with a self-developed multi-mode audio frequency pulse photoelectric therapeutic apparatus), acupuncture group (Group 2), and oral placebo group (Group 3), 58 cases per group. The curative effect of each group was evaluated by the changes of blood pressure and CM syndrome integral before and after treatment. RESULTS: Compared with Group 3, there were significant decrease of blood pressure and CM syndrome integral in Group 1 and Group 2 (P<0.01). Compared with Group 2, Group 1 showed the highest decrease in systolic pressure (P<0.017). The total effective rate of anti-hypertension in Group 1 (91.38%, 53/58) was significantly higher than that in Group 2 (74.13%, 43/58) and Group 3 (18.97%, 11/58, P<0.05 or P<0.01); and that in Group 2 was also significantly higher than that in Group 3 (P<0.01). There were significant difference in the total effective rate of CM syndrome integral in both Group 1 (93.10%, 54/58) and Group 2 (84.48%, 49/58) as compared with Group 3 (17.24%, 10/58, P<0.01), while there was no significant difference between Group 1 and Group 2 (P>0.05). CONCLUSIONS: The multi-mode audio frequency pulse photoelectric therapeutic apparatus, combining music, laser and electric pulse stimulation, is clinically useful for grade 1 essential hypertension. This "three in one" therapy method is non-invasive, easy and simple to handle. It is expected to be popularized as a new alternative treatment.


Assuntos
Pontos de Acupuntura , Terapia por Estimulação Elétrica , Hipertensão/terapia , Lasers , Música , Pressão Sanguínea , Hipertensão Essencial , Feminino , Humanos , Hipertensão/fisiopatologia , Masculino , Medicina Tradicional Chinesa , Pessoa de Meia-Idade , Síndrome , Resultado do Tratamento
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