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2.
Anal Bioanal Chem ; 412(25): 7007-7016, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32740822

RESUMO

Qualitative and quantitative detection of genetically modified products is inseparable from the application of reference materials (RMs). In this study, a batch of genomic DNA (gDNA) certified reference materials (CRMs) was developed using genetically modified rice Kemingdao (KMD) homozygotes as the raw material. The gDNA CRMs in this batch showed good homogeneity; the minimum sample intake was determined to be 2 µL. The stability study showed that transportation by cold chain is preferable, no significant degradation trend was observed during a 12-month period when storing the gDNA CRMs at 4 °C and - 20 °C, and the number of freeze-thaw cycles cannot exceed 10. The property values of the copy number ratio of transgene and endogenous gene and the copy number concentration for gDNA CRMs were determined by a collaborative characterization of eight laboratories using the duplex KMD/PLD droplet digital PCR (ddPCR) assays. The uncertainty components of characterization, potential between-unit heterogeneity, and potential degradation during long-term storage were combined to estimate the expanded uncertainty of the certified value with a coverage factor k of 2.0. The certified value of copy number ratio for KMD gDNA CRM is 0.99 ± 0.05, and that of copy number concentration is (1.76 ± 0.10) × 105 copies/µL. Compared to the gDNA CRMs in availability, this batch of KMD gDNA CRMs is assigned accurate property values and can be directly used for qualitative and quantitative detection of GMOs as well as evaluation of the parameters of analytical methods with no need of further DNA concentration measurement. Graphical abstract.

3.
Zhongguo Fei Ai Za Zhi ; 23(8): 662-666, 2020 Aug 20.
Artigo em Chinês | MEDLINE | ID: mdl-32752579

RESUMO

BACKGROUND: How to locate pulmonary ground-glass nodules in thoracoscopic surgery is an important clinical topic in minimally invasive thoracic surgery. There is no unified localization method at present. This study intends to investigate the accuracy and security of body surface theodolitic puncture localization method in video-assisted thoracoscopic surgery for pulmonary ground-glass nodules. METHODS: The clinical data of 41 patients from August 2018 to December 2019 were analyzed retrospectively, including 28 males and 13 females. After anesthesia, the patient was located by body surface theodolitic puncture, and then partial lobectomy was performed under video-assisted thoracoscopy. The distance from the nodule to the marked suture and the distance from the nodule to the incisal margin were measured, and the accuracy of localization, the rate of complication and the success rate of surgical resection were calculated. RESULTS: A total of 51 nodules in 41 patients were located by body surface theodolitic puncture localization method. The accuracy rate was 96.1%, and the average location time was 8.3 min. Puncture bleeding occurred in 5 cases (12.2%), all of which were successfully stopped by video-assisted thoracoscopy, and there were no other complications. All patients underwent thoracoscopic partial lobectomy, including 33 cases of anatomical segmentectomy and 8 cases of wedge lobectomy. All the patients in operation process smoothly. The distance between nodule and incisal margin was measured, and all specimens were more than 2 cm, reaching a safe distance. The success rate of surgical resection was 100.0%. CONCLUSIONS: In video-assisted thoracoscopic surgery for ground glass nodules of lung, the body surface theodolitic puncture localization method can be accurate, safe and simple.

4.
Biochem Biophys Res Commun ; 528(3): 440-446, 2020 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-32507599

RESUMO

Previous studies have shown that the occurrence of atherosclerosis is closely related to changes of α2, 6-sialic acid transferase I (ST6Gal-I). Bace1 has been identified as a protease responsible for the cleavage and secretion of Golgi-resident ST6Gal-I. There have been only a few attempts to clarify the direct connection between Bace1 and atherosclerosis. The purpose of this study was to investigate the relationship between Bace1 gene and atherosclerosis. Expressions of Bace1 protein and mRNA in ApoE-/- mice fed on high-fat diet were evaluated and the development of atherosclerosis was assessed in Bace1-/- mice fed on high-fat diet. In vitro, the expression of Bace1 gene was detected in foam cell model and the formation of foam cells was examined after knocking down Bace1 by siRNA. We observed a significant increase in Bace1 expression in the aortic root in the model of atherosclerosis in ApoE-/-mice. The expression of Bace1 protein and mRNA levels had a remarkable increase in high-fat group. After knocking out the Bace1 gene, serum lipid levels were significantly lower and intimal thickness was obvious thinner than those in wild-type mice with high-fat diet. Expression of Bace1 protein and mRNA levels were significantly elevated in foam cell. The formation of foam cells was blocked when Bace1 was knocked down by siRNA interferes. Our results suggested that elevated Bace1 gene had a positive role in the progression of atherosclerosis. Affecting the glycosyltransferase may be one of its mechanisms.

5.
Biol Chem ; 2020 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-32386184

RESUMO

It has been reported that high-mobility group box 3 is overexpressed in various cancers. This study aimed to explore its function in non-small cell lung cancer (NSCLC). A546 and H460 cell lines were used for in vivo experiments, scratch healing tests, transwell migration and invasion experiments. It was first found that HMGB3 was highly expressed in tumor tissues in the patients and associated with NSCLC stage. Silencing of HMGB3 significantly slowed the growth, proliferation and invasion of NSCLC in vitro, and repressed cell growth in vivo. Mechanistic studies suggest that the observed effects were mediated by inhibiting the expression of ß-catenin/MMP7/c-Myc in Wnt pathway. Our study highlights the role of HMGB3 in NSCLC, which may provide a therapeutic target for the treatment of NSCLC.

6.
Immunol Lett ; 222: 67-72, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32197974

RESUMO

To develop anti-tumor agents for lung cancer, we aim to characterize a herbal compound, daidzein-rich isoflavones aglycone (DRIA), in inhibiting the proliferation and NF-κB signaling pathway of lung cancer. MTT and colony formation assays were used to analyze the proliferation of lung cancer cells in presence of DRIA treatment, which showed that DRIA dose-dependently inhibited the proliferation and colony formation of lung cancer cells. Enzyme-linked immunosorbent assay revealed that interleukin-6 (IL6) and interleukin-8 (IL-8) levels were reduced by DRIA. p65-NFκB expression and activation, which was enhanced by TNF-α and C/EBPß treatment, were attenuated by DRIA. Exogenous tumor necrosis factor-α (TNF-α) and CCAAT/enhancer binding protein (C/EBPß) were used to enhance NF-κB signaling in cells, and the effects of DRIA in attenuating NF-κB signaling were assessed by analyzing p65-NFκB expression in mRNA and protein levels, using quantitative real-time PCR (qRT-PCR), western blot and immunofluorescence staining. immunohistochemical staining revealed that Ki-67 and p65-NF-κB levels in A594 tumor xenografts of A594 tumors were also reduced by DRIA treatment in mice. Our data indicates that DRIA is effective in inhibiting the proliferation and NFκB signaling of lung cancer both in vitro and in vivo.

7.
Acta Biochim Biophys Sin (Shanghai) ; 52(4): 371-381, 2020 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-32188965

RESUMO

As a subtype of non-small-cell lung cancer, lung squamous cell carcinoma (LUSC) accounts for one-fifth of all lung cancers. Unfortunately, no specific targetable aberration has yet been identified. Hence, it is of huge urgency and potential to identify aberrantly regulated genes in LUSC. Here, five pairs of LUSC samples and their corresponding adjacent tissues were subject to whole transcriptome sequencing. Our results showed that CTD-2562J17.6 and FENDRR were significantly downregulated while MIR205HG, LNC_000378, RP11-116G8.5, RP3-523K23.2, and RP5-968D22.1 were significantly upregulated in all five LUSC samples. Importantly, MIR205HG was upregulated in LUSC clinical samples as well as in LUSC cell lines. Interestingly, our results demonstrated that the expression level of MIR205HG is positively correlated with the malignancy. In addition, MIR205HG is required for LUSC cell growth and cell migration. Most importantly, our results showed that MIR205HG prohibits LUSC apoptosis via regulating Bcl-2 and Bax. Taken together, our data shed lights on the lncRNA regulatory nexus that controls the carcinogenesis of LUSC and provided potential novel diagnostic markers and therapeutic targets for LUSC.

8.
Arch. bronconeumol. (Ed. impr.) ; 55(12): 627-633, dic. 2019. graf
Artigo em Espanhol | IBECS | ID: ibc-186396

RESUMO

Introduction: Lung cancer is a major public health problem, as the second causes of cancer related death worldwide, with relatively low survival rates, and accessible drug resistance. Long non-coding RNAs (LncRNAs) have been identified as activator in lung cancer with elusive mechanisms. We aimed to detect the regulation of LncRNA MALAT1 in the proliferation and gefitinib resistance in lung cancer cells. Methods: MALAT1 in A549 and HCC 1299 human lung adenocarcinoma cell lines was silenced by shRNA or overexpressed using plasmid, and the cell viability and cell proliferation were evaluated by MTT assay and soft agar colony formation assay. RNA levels were detected by RT-PCR, and the protein expression was measured by western blot. The binding between MALAT1 and miR-200a was validated by luciferase reporter assays using pSi-Chech 2 vectors. Results: The cell viability and proliferation of A549 cells transfected with MALAT1 shRNA were significantly lower than the control. The MALAT1 expression in gefitinib resistant A549 cells was upregulated. miR-200a significantly inhibited the fluorescence of pSi-Check 2 vector with MALAT1 gene, suggesting the direct binding between MALAT1 and miR-200a. In addition, LncRNA MALAT1 promotes ZEB1 expression in A549 cells. Conclusion: Our study showed that MALAT1 promoted the proliferation and gefitinib resistance of lung cancer cells by sponging miR-200a, which regulates expression of ZEB1 in the A549 cells. This MALAT1/miR-200a axis could serve as new therapeutic target for lung cancer treatment


Introducción: El cáncer de pulmón es un importante problema de salud pública. Constituye la segunda causa de muerte por cáncer en el mundo y se asocia a tasas de supervivencia relativamente bajas y a resistencia a fármacos accesibles. Los RNA largos no-codificantes (lncRNA) se han identificado como activadores en el cáncer de pulmón con mecanismos aún no esclarecidos. El objetivo de este estudio fue detectar la regulación del LncRNA MALAT1 en la proliferación y la resistencia de las células tumorales de pulmón. Métodos: La expresión de MALAT1 se silenció con un shRNA o se sobreexpresó mediante el uso de un plásmido en las líneas celulares de adenocarcinoma pulmonar humano A459 y HCC. La viabilidad celular y la proliferación se evaluaron mediante un ensayo MTT y el ensayo de formación de colonias en agar blando. Los niveles de RNA se detectaron con RT-PCR y los niveles de proteína se midieron con Western Blot. La unión entre MALAT1 y miR-200a se validó gracias a un ensayo de luciferasa utilizando vectores pSi-Chech 2. Resultados: La viabilidad y la proliferación de las células A549 transfectadas con el shRNA contra MALAT1 resultaron significativamente más bajas que en el control. Se produjo un incremento en la expresión de MALAT1 en las células A549 resistentes a gefitinib. MiR-200a inhibió significativamente la fluorescencia del vector pSi-Check 2 que contenía el gen MALAT1, sugiriendo la unión directa entre MALAT1 y miR-200a. Además, el LncRNA de MALAT1 promovió la expresión de ZEB1 en las células A549. Conclusión: Nuestro estudio demuestra que MALAT1 promovió la proliferación y resistencia a gefitinib en células tumorales de pulmón actuando como "esponja" de miR-200a, que regula la expresión de ZEB1 en células A549


Assuntos
Humanos , MicroRNAs/genética , Proteínas de Ligação a RNA/genética , RNA Longo não Codificante/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/diagnóstico , Resistência a Medicamentos/efeitos dos fármacos , Adenocarcinoma/diagnóstico , Gefitinibe/uso terapêutico , Reação em Cadeia da Polimerase
9.
Artif Cells Nanomed Biotechnol ; 47(1): 3904-3912, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31566021

RESUMO

This study aimed to investigate the effect of Junduqing extractive on proliferation, apoptosis, migration and invasion of nasopharyngeal carcinoma (NPC) cells and the involved mechanism. Junduqing extractive was prepared. CCK-8 assay found that IC50 of Junduqing extractive in HNE-1 cells was 2.99 mg/ml, so its concentration of 1.0, 2.0 and 3.0 mg/ml was selected to perform the following experiments. HNE-1, HNE-2 and HONE1 cells were then divided into four groups: (1) Control (no treatment); (2) 1.0 mg/ml (1.0 mg/ml Junduqing); (3) 2.0 mg/ml (2.0 mg/ml Junduqing) and (4) 3.0 mg/ml (3.0 mg/ml Junduqing). Cell viability, apoptosis, migration and invasion were examined by CCK-8 assay, annexin V-FITC/PI staining, scratch wound assay and transwell assay, respectively. Compared with the control group, the viability, migration rates and invasive capacity of HNE-1, HNE-2 and HONE1 cells with Junduqing treatments decreased significantly. Higher concentration of Junduqing extractive caused lower viability, smaller migration rates and weaker invasive capacity. Compared with the control group, the apoptosis of HNE-1, HNE-2 and HONE1 cells after treatment with 2.0 and 3.0 mg/ml of Junduqing extractive increased remarkably. Levels of Bcl-xL, Mcl-1, Caspase-3, Caspase-8 and Caspase-9 were examined by western blotting. Compared with the control group, the expression of Bcl-xL and Mcl-1 and the expression of Caspase-3, Caspase-8 and Caspase-9 in HNE-1, HNE-2 and HONE1 cells were significantly down-regulated and up-regulated, respectively, after treatment with Junduqing extractive. In conclusion, Junduqing extractive could inhibit the proliferation, migration and invasion, and promote the apoptosis of human NPC cells through down-regulating Mcl-1 and Bcl-xL and up-regulating Caspase-3, Caspase-8 and Caspase-9.


Assuntos
Apoptose/efeitos dos fármacos , Caspases/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Carcinoma Nasofaríngeo/patologia , Proteína bcl-X/metabolismo , Antineoplásicos Fitogênicos/farmacologia , Caspase 3/metabolismo , Caspase 8/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Humanos , Invasividade Neoplásica , Regulação para Cima/efeitos dos fármacos
10.
Mol Cell Biochem ; 460(1-2): 1-8, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31187349

RESUMO

Lung cancer is the major cause leading to cancer mortality, and the 5-year survival rate for patients with lung cancer still remains low. It is urgent to fully understand the development and progression of lung cancer to discover new therapeutic targets and develop new therapeutic approaches. H19 was documented to be upregulated in lung cancer and related to cell proliferation. However, it is still unclear if H19 has other functions in lung cancer. The mRNA levels of genes were detected by qRT-PCR, and the cell proliferation rate and cell viability were measured through cell count assay and MTT assay. Transwell assays were applied to detect cell abilities to migration and invasion, while luciferase reporter assay and RNA pull-down assay were used to examine interaction between H19 and miR-200a. H19 expression was elevated in the lung cancer tissues and cell lines, while H19 overexpression promoted the lung cancer cell growth, cell migration and invasion, as well as the epithelial mesenchymal transition (EMT). Meantime, RNA pull-down assay showed that H19 interacted with miR-200a, and miR-200a inhibited the activity of H19-fused luciferase. Furthermore, H19 overexpression inhibited miR-200a function and thereby upregulated miR-200a target genes, ZEB1 and ZEB2.H19 sponged and inhibited miR-200a to de-repress expression of ZEB1 and ZEB2, and thereby enhanced lung cancer proliferation and metastasis.


Assuntos
Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs/genética , Invasividade Neoplásica , RNA Longo não Codificante/genética , Regulação para Cima/genética , Homeobox 2 de Ligação a E-box com Dedos de Zinco/genética , Homeobox 2 de Ligação a E-box com Dedos de Zinco/metabolismo , Homeobox 1 de Ligação a E-box em Dedo de Zinco/genética , Homeobox 1 de Ligação a E-box em Dedo de Zinco/metabolismo
11.
Arch Bronconeumol ; 55(12): 627-633, 2019 12.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-31133357

RESUMO

INTRODUCTION: Lung cancer is a major public health problem, as the second causes of cancer related death worldwide, with relatively low survival rates, and accessible drug resistance. Long non-coding RNAs (LncRNAs) have been identified as activator in lung cancer with elusive mechanisms. We aimed to detect the regulation of LncRNA MALAT1 in the proliferation and gefitinib resistance in lung cancer cells. METHODS: MALAT1 in A549 and HCC 1299 human lung adenocarcinoma cell lines was silenced by shRNA or overexpressed using plasmid, and the cell viability and cell proliferation were evaluated by MTT assay and soft agar colony formation assay. RNA levels were detected by RT-PCR, and the protein expression was measured by western blot. The binding between MALAT1 and miR-200a was validated by luciferase reporter assays using pSi-Chech 2 vectors. RESULTS: The cell viability and proliferation of A549 cells transfected with MALAT1 shRNA were significantly lower than the control. The MALAT1 expression in gefitinib resistant A549 cells was upregulated. miR-200a significantly inhibited the fluorescence of pSi-Check 2 vector with MALAT1 gene, suggesting the direct binding between MALAT1 and miR-200a. In addition, LncRNA MALAT1 promotes ZEB1 expression in A549 cells. CONCLUSION: Our study showed that MALAT1 promoted the proliferation and gefitinib resistance of lung cancer cells by sponging miR-200a, which regulates expression of ZEB1 in the A549 cells. This MALAT1/miR-200a axis could serve as new therapeutic target for lung cancer treatment.

12.
Anal Bioanal Chem ; 411(9): 1729-1744, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30707265

RESUMO

The enforcement of GMO labeling regulations requires validated analytical methods and certified reference materials (CRMs). The early labeling regulations stipulated that the GMO content should be expressed as percentage, but did not specify what unit this percentage referred to. Two reference systems, using mass fraction and copy number ratio as measurement units, individually, are established for GMO analysis using different metrological traceability chains. Three types of CRMs, powder CRMs certified for mass fractions, genomic DNA CRMs, and plasmid DNA CRMs certified for copy number ratios, were developed for calibration and quality control. The type, certification, and measurement unit commutability of current GMO CRMs are presented and discussed in this paper. Both existing reference systems are facing a metrological challenge, although later EU regulations specified that the measurement unit of GMO content must be expressed in mass fraction and recommended to convert one unit into another by introducing a conversion factor, further efforts are required to explore which reference system is more metrologically sound. The determination of conversion factor per CRM batch is recommended to be based on the pure CRMs produced from pure GM materials, which is expected to be the best choice for calibration of PCR measurement results.


Assuntos
DNA de Plantas/genética , Organismos Geneticamente Modificados , Genes de Plantas , Limite de Detecção
13.
Surg Innov ; 26(3): 337-343, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30694104

RESUMO

OBJECTIVES: Video-assisted thoracoscopic surgery (VATS) pulmonary segmentectomy is commonly used in treating small ground-glass opacity (GGO) nodules in lung. The identification of the intersegmental plane is one of the challenges. In this pilot study, we aimed to evaluate the feasibility of indocyanine green (ICG) angiography in VATS segmentectomy. METHODS: Nineteen GGO patients were enrolled, and VATS segmentectomy with ICG near-infrared angiography were performed between July 2017 and December 2017. Conventional 3-port VATS was used. ICG was injected intravenously after dominant arties were ligated. Intersegmental plane was identified and divided by stapler and electrocautery. RESULTS: All patients had perfect intersegmental plane visualization. The mean operation time was 140.8 minutes, and the mean blood loss was 23.7 mL. No complications due to ICG occurred. The mean chest tube duration was 4.6 days. No severe complications occurred in the perioperative period. The mean chest tube drainage duration was 4.6 days. Prolonged postoperative air leak (>5 days), which required no surgical intervention, occurred in 2 cases. There were no severe complications or in-hospital deaths. CONCLUSIONS: VATS segmentectomy with ICG near-infrared angiography is a reasonable treatment option to treat small GGO in lung, especially due to its good surgical view maintenance.


Assuntos
Angiografia/métodos , Verde de Indocianina/administração & dosagem , Neoplasias Pulmonares/cirurgia , Pneumonectomia/métodos , Cirurgia Torácica Vídeoassistida , Perda Sanguínea Cirúrgica/estatística & dados numéricos , Tubos Torácicos/estatística & dados numéricos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Projetos Piloto
14.
Afr Health Sci ; 19(3): 2526-2536, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32127825

RESUMO

Background: Erythrina variegata has been widely used as a traditional medicine. Objective: The study was designed to evaluate the anxiolytic and anti-depressant effects of an extract from Erythrina variegata. Methods: The extract was evaluated for anxiolytic and anti-depressant action using the elevated plus maze, light/dark box, open field, forced swimming and tail suspension tests in mice. The mechanism of action was further elucidated using high-performance liquid chromatography with fluorescence detection methods to assay the levels of five neurotransmitters in brain. Results: The extract exhibited significant increase in the percentage of the open arms entries and the time spent in the open arms in the elevated plus maze test. The results of the light/dark box test revealed a significant increase in the amount of time spent in the light chamber. Extract- treated mice also produced significant increase in the number of crossings and rearings in the open field test. In the forced swimming and tail suspension tests, the extract was able to promote significant decrease in the immobility time. In addition, the extract significantly altered the levels of five neurotransmitters in the brain tissue. Conclusion: These findings suggest that Erythrina variegata presents potential anxiolytic and anti-depressant activity, and the mechanism may be related to the alteration of neurotransmitter levels.

15.
Neural Regen Res ; 13(7): 1225-1230, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30028331

RESUMO

Chronic stress is strongly associated with the occurrence and development of depression and cardiovascular disease. Stress can induce altered mitochondrial function and activation of apoptosis in the cardio-cerebral system. However, it is unknown whether the protein kinase C ε (PKCε)-aldehyde dehydrogenase 2 (ALDH2) pathway is altered under chronic stress, and this study sought to address this question. A rat model of depression was established using a chronic unpredictable mild stress (CUMS) protocol. After experiencing CUMS for 4 weeks, the sucrose preference test and the forced swim test verified depressive-like behaviors. Enzyme linked immunosorbent assays showed that ALDH2 activity was decreased in the rat hippocampus and prefrontal cortex, but was not altered in the myocardium. Western blot assays demonstrated reduced levels of ALDH2 and PKCε, but increased levels of 4-hydroxy-2-nonenal (4HNE) adducts. Caspase-3 expression did not obviously alter, but active forms of caspase-3 were increased in the hippocampus and prefrontal cortex. In the myocardium, expression of ALDH2, PKCε and 4HNE adducts did not remarkably alter; while caspase-3 expression was reduced and the active forms of caspase-3 were upregulated. Pearson's correlation test demonstrated that expression of 4HNE adducts was positively correlated with levels of the active forms of caspase-3 in the hippocampus and prefrontal cortex, but not in the myocardium. In conclusion, chronic stress can damage the PKCε-ALDH2 signaling pathway in the hippocampus and prefrontal cortex, but not in the myocardium. Moreover, 4HNE is associated with active forms of caspase-3 in the hippocampus and prefrontal cortex.

16.
Zhongguo Zhong Yao Za Zhi ; 42(21): 4115-4119, 2017 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-29271148

RESUMO

By determination the section color and lustre indexes as well as the content of baicalin in 30 batches of Scutellariae Radix slices, calculate the correlation of these two, screen the color and lustre indexes which could represent their intrinsic quality, and establish a new grade classification method based on the results. The results showed that samples met the conditions of △L≥-37, △b≥45 simultaneously were picked grade and the content of baicalin was of ≥200 mg•g⁻¹ definitely; Samples inconsistent with any one of above conditions were general grade. This research indicated that indexes of △L and △b could characterize both the color and luster of slice and intrinsic quality, so that could be used as the indexes to classify the grades of Scutellariae Radix slices accurately, easily and objectively. The research results would provide new ideas and references for grade classification of traditional Chinese medicine slices.


Assuntos
Medicamentos de Ervas Chinesas/normas , Raízes de Plantas/química , Scutellaria baicalensis/química , Medicamentos de Ervas Chinesas/química , Flavanonas/análise , Medicina Tradicional Chinesa
17.
J Sci Food Agric ; 97(5): 1634-1639, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27436567

RESUMO

BACKGROUND: As event-specific sequence information for most unauthorised genetically modified organisms (GMOs) is currently still unavailable, detecting unauthorised GMOs remains challenging. Here, we used insect-resistant rice TT51-1 as an example to develop a novel approach via detecting GMOs by RNA-seq (sequencing) and PCR. RNA-seq of TT51-1 generated 4.8 million (M) 21-nt cDNA tags. Alignment to the Oryza sativa subsp. japonica reference genome revealed 24 098 unmapped tags. Foreign tags from the nopaline synthetic enzyme gene (NOS) terminator and insect-resistant genes were then identified by searching against the NCBI VecScreen and NT databases. RESULTS: To further isolate foreign DNA sequences, putative NOS terminator and insect-resistant gene tags were combined and used directly as primer pairs for long-range PCR, producing a 5016-bp fragment. The inserted DNA sequence of TT51-1 has been submitted to a database, and thus, similarity analysis using the database could identify a test sample. CONCLUSION: The novel approach has a great potential for application to the detection and identification of unauthorised GMOs in food and feed products. © 2016 Society of Chemical Industry.


Assuntos
Oryza/genética , Plantas Geneticamente Modificadas/genética , Agrobacterium tumefaciens/genética , Aminoácido Oxirredutases/genética , Bacillus thuringiensis/genética , Bases de Dados de Ácidos Nucleicos , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA , Regiões Terminadoras Genéticas
18.
Sci Rep ; 6: 33955, 2016 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-27670217

RESUMO

Genic male sterility (GMS) has already been extensively utilized for hybrid rapeseed production. TE5A is a novel thermo-sensitive dominant GMS line in Brassica napus, however, its mechanisms of GMS remain largely unclear. Histological and Transmission electron microscopy (TEM) analyses of anthers showed that the male gamete development of TE5A was arrested at meiosis prophase I. EdU uptake of S-phase meiocytes revealed that the TE5A mutant could accomplish DNA replication, however, chromosomal and fluorescence in situ hybridization (FISH) analyses of TE5A showed that homologous chromosomes could not pair, synapse, condense and form bivalents. We then analyzed the transcriptome differences between young floral buds of sterile plants and its near-isogenic fertile plants through RNA-Seq. A total of 3,841 differentially expressed genes (DEGs) were obtained, some of which were associated with homologous chromosome behavior and cell cycle control during meiosis. Dynamic expression changes of selected candidate DEGs were then analyzed at different anther developmental stages. The present study not only demonstrated that the TE5A mutant had defects in meiotic prophase I via detailed cytological analysis, but also provided a global insight into GMS-associated DEGs and elucidated the mechanisms of GMS in TE5A through RNA-Seq.

19.
Eur J Pharm Sci ; 93: 224-32, 2016 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-27493021

RESUMO

Water-soluble Cistanche phenylethanoid glycosides (CPhGs) have poor permeability and low bioavailability. However, liposomes can improve the permeability of such drugs and their poor stability, and proliposomes have been used to overcome these problems. Based on this, Cistanche phenylethanoid glycoside liquid proliposomes (CPhGsP) and dripping(?) pills were prepared and optimized using response surface methodology. The properties of CPhGsP were evaluated in terms of their encapsulation efficiency, particle size, zeta potential, and morphology. The results obtained showed that the optimal formulation was drug/soybean phospholipid/poloxamer-188/sodium deoxycholate/propylene glycol 1:22.38:3.52:0.84:80 (w/w/w/w/v). This resulted in an encapsulation efficiency, particle size, and zeta potential of hydrated proliposomes with phosphate buffer solution (pH7.4) of 51.97%, 671.7nm, and -25.49mV, respectively. Stability testing of CPhGsP and CPhGs ordinary liposomes was carried out for 3months at 4±2°C, 25±2°C, 40±2°C, 75±5% RH. The results obtained showed that the stability of the proliposomes was better than that of ordinary liposomes at the same temperature, while a lower temperature of 4°C is ideal for storage. Cistanche phenylethanoid glycoside liquid proliposomes dripping pills (CPhGsPD) are efficiently released in gastrointestinal solution as shown by in vitro release experiments and the structure of the liposomes does not destroy the proliposome dripping pills by hydration. In vivo experiments showed that the areas under the plasma level-time curves and peak concentrations of CPhGsPD and hydrated proliposomes were higher than those of CPhGs. Moreover, with CPhGsPD, the pharmacokinetic parameters were similar to those with hydrated proliposomes. These results showed that CPhGsPD offer a good way to improve the oral delivery of CPhGs.


Assuntos
Cistanche , Glicosídeos/administração & dosagem , Administração Oral , Animais , Disponibilidade Biológica , Ácido Desoxicólico/química , Composição de Medicamentos , Glicosídeos/química , Glicosídeos/farmacocinética , Lipossomos , Masculino , Fosfolipídeos/química , Poloxâmero/química , Propilenoglicol/química , Coelhos
20.
Plasmid ; 87-88: 28-36, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27497661

RESUMO

The use of analytical controls is essential when performing GMO detection through screening tests. Additionally, the presence of taxon-specific sequences is analyzed mostly for quality control during GMO detection. In this study, 11 commonly used genetic elements involving three promoters (P-35S, P-FMV35S and P-NOS), four marker genes (Bar, NPTII, HPT and Pmi), and four terminators (T-NOS, T-35S, T-g7 and T-e9), together with the reference gene fragments from six major crops of maize, soybean, rapeseed, rice, cotton and wheat, were co-integrated into the same single plasmid to construct a general reference plasmid pBI121-Screening. The suitability test of pBI121-Screening plasmid as reference material indicated that the non-target sequence on the pBI121-Screening plasmid did not affect the PCR amplification efficiencies of screening methods and taxon-specific methods. The sensitivity of screening and taxon-specific assays ranged from 5 to 10 copies of pBI121-Screening plasmid, meeting the sensitivity requirement of GMO detection. The construction of pBI121-Screening solves the lack of a general positive control for screening tests, thereby reducing the workload and cost of preparing a plurality of the positive control.


Assuntos
Plantas Geneticamente Modificadas/genética , Plasmídeos/genética , Padrões de Referência , Código de Barras de DNA Taxonômico , Genes de Plantas , Plantas Geneticamente Modificadas/classificação , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade
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