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OBJECTIVE: To examine the expression and function of necroptosis-associated miRNAs in esophageal squamous cell carcinoma. METHODS: A total of three microarray datasets, i.e., GSE122497, GSE114110, and GSE43732, were selected from the GEO database for differential analysis of necroptosis-related miRNA expression. The differentially expressed miRNAs were screened for target miRNAs using Kaplan-Meier survival analysis in the OncomiR database. The expression of the target miRNAs in the HEEC, KYSE-450, TE-1, and KYSE-410 cell lines was measured via qPCR. The expression of the target miRNAs in esophageal cancer cells was regulated by transfection with Lipofectamine 2000, and cell proliferation, cell migration, cell apoptosis and the cell cycle were detected by CCK-8, Transwell, and flow cytometry. RESULTS: The tumor tissue and peripheral blood of esophageal squamous cell cancer patients showed differential expression of 7 miRNAs related to necroptosis. Survival analysis revealed that miR-425-5p and miR-16-5p were negatively correlated with patient survival. The esophageal squamous cell carcinoma cell lines exhibited increased expression of miR-425-5p and miR-16-5p, with KYSE-410 exhibiting the most significant increase. Inhibition of miR-425-5p and miR-16-5p expression in the KYSE-410 cell line resulted in increased apoptosis, decreased proliferation, and decreased migration of esophageal cancer cells as well as a significant increase in the S phase and a decrease in the G2/M phase according to the cell cycle assay. CONCLUSION: The pro-carcinogenic role of miR-425-5p and miR-16-5p has been observed in esophageal squamous cell carcinoma.
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Neoplasias Esofágicas , Carcinoma de Células Escamosas de Esófago , MicroARNs , Humanos , Carcinogénesis/genética , Carcinógenos , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/patología , Carcinoma de Células Escamosas de Esófago/genética , Carcinoma de Células Escamosas de Esófago/metabolismo , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , MicroARNs/metabolismo , Necroptosis/genéticaRESUMEN
Recently, hip-hop pedagogy or Hip-Hop Based Education (HHBE) have become buzz words in the academic and public debate around hip-hop. However, we found that most definitions of hip-hop pedagogy are missing the concept of pedagogy itself. One consequence of failing to adequately explain the concept of pedagogy is that it may lead future hip-hop researchers, students, and teachers inadvertently to disseminate misinformation or foster unclear thinking by using "hip-hop pedagogy" in inaccurate or vague ways. For these reasons, it is important to have a shared understanding of hip-hop pedagogy. In this article, we present three updated, expanded definitions of hip-hop pedagogy with the potential for widespread acceptance. These definitions aim to convey in the simplest terms what hip-hop pedagogy is for the purpose of informing educators and preparing them to use data.
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The piggery digestate of high ammonia was mixed with the anoxic aerated effluent of high nitrate and phosphorus, to cultivate a microalgal-bacterial consortium for simultaneous pollution removal and resource recovery. The highest removal of total inorganic nitrogen was achieved at 324.77 mg/L in 40% piggery digestate mixed with 60% anoxic aerated effluent, along with the most microalgae biomass production. The crude protein and fatty acids of C14-C20 in microalgae cells were 21.80% and 69.78%, indicating that this mixing strategy could produce abundant microalgal biomass suitable for biofuel generation and animal feed. High-throughput sequencing showed that microbial diversity increased and Paenibacillus, Thiopseudomonas and Pseudomonas were the dominant species promoting microalgal growth. Overall, these results provided a new insight of mixing two types of wastewaters for cultivating microalgal-bacterial consortia, to remove contamination and recover nutrients simultaneously.
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Microalgas , Animales , Biomasa , Nitrógeno , Fósforo , Aguas ResidualesRESUMEN
OBJECTIVE: To observe the clinical effect of acupuncture therapy in the treatment of postpartum obesity subjects. METHODS: A total of 71 postpartum obesity subjects were allocated to treatment group (n=35) and control group (n=36). Participants of the control group were asked to receive weekly nutrition consultation (body-weight management) for calorie intake control during 4 weeks' treatment, and those of the acupuncture group were treated by manual acupuncture stimulation of Zhongwan (CV 12), Zhongji(CV 3), Qihai(CV 6), Shuifen (CV 9), and Guanyuan (CV 4), bilateral Tianshu (ST 25), Guilai (ST 29), Shousanli (LI 10), Zusanli (ST 36), Fenglong (ST 40), and Yinlingquan (SP 9) in combination with nutrition consultation. The treatment was conducted once every other day, continuously for 4 weeks. The body weight(BW), body mass index (BMI), percent of body fat (PBF) and waist-to-hip ratio (WHR) were determined before and after the treatment. RESULTS: After the treatment, the BW, BMI, PBF and WHR in the acupuncture group, and the BW and WHR in the control group were significantly decreased (P<0.01, P<0.05), and the therapeutic effect of the acupuncture group was notably superior to that of the control group in reducing BW and WHR (P<0.05, P<0.01). CONCLUSIONS: Acupuncture therapy combined with nutritional calorie control has a positive role in relieving obesity in postpartum obesity participants.
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Terapia por Acupuntura , Obesidad/terapia , Periodo Posparto , Pérdida de Peso , Puntos de Acupuntura , Dieta Reductora , Femenino , HumanosRESUMEN
A Gram-stain-negative, rod-shaped and motile bacterial strain, designated A9T, was isolated from the surface of rock collected from the shore of Nvshan lake in Mingguang, Anhui province, China. Phylogenetic analysis based on 16S rDNA sequence data showed that strain A9T was affiliated with the genus Massilia and showed the highest sequence similarities to Massilia plicata KCTC 12344T (98.8â%) and Massilia lurida CGMCC 1.10822T (97.9â%). The major fatty acids (>5â%) were summed feature 3 (C16â:â1ω7c and/or C15â:â0 iso 2-OH), C16â:â0 and C18â:â1ω7c. Strain A9T contained Q-8 as the predominant ubiquinone and diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and an unidentified aminophospholipid as the predominant polar lipids. The DNA G+C content was 69.9 mol%. Mean DNA-DNA relatedness values between strain A9T and its closest phylogenetic relatives, M. plicata KCTC 12344T and M. lurida CGMCC 1.10822T, were 38.8â% and 23.23â%, respectively. On the basis of the results obtained in this study, strain A9T is considered to represent a novel species of the genus Massilia, for which the name Massilia buxea sp. nov. is proposed. The type strain is A9T (=DSM 103547T=CGMCC 1.15931T=KCTC 52429T).
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Oxalobacteraceae/clasificación , Filogenia , Técnicas de Tipificación Bacteriana , Composición de Base , China , ADN Bacteriano/genética , Ácidos Grasos/química , Hibridación de Ácido Nucleico , Oxalobacteraceae/genética , Oxalobacteraceae/aislamiento & purificación , Fosfolípidos/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Ubiquinona/químicaRESUMEN
AIM:To investigate the protective effect of curcumin analogue L 6H4 on diaphragm of type 2 dia-betic rats.METHODS: SPF male Sprague-Dawley rats ( n=40) were randomly divided into 5 groups: normal control (NC) group, high fat (HF) group, high fat+L6H4 treatment (FT) group, diabetes mellitus (DM) group and DM +L6H4 treatment (DT) group.The rats in the later 4 groups were fed with high-fat diet.After 4 weeks of high-fat diet fee-ding, the rats in DM and DT groups were intraperitoneally injected with streptozotocin to induce type 2 diabetes melliutus. The rats in FT and DT groups were given L6H4 by gavage for 8 weeks.Blood glucose and blood lipid levels were detected biochemically .Fasting serum insulin ( FINS ) level was measured by radioimmunoassay and insulin resistance index (HOMA-IR) was calculated.Serum adiponectin (APN) level was measured by ELISA.The morphological changes of the diaphragm were observed under light and transmission electron microscopes .Lipid deposition and the activity of succinate dehydrogenase (SDH) and NADH-tetrazolium reductase (NADH-TR) were observed by enzyme histochemical staining . The content of malondialdehyde ( MDA) and the activity of superoxide dismutase ( SOD) in the diaphragm were measured by thiobarbituric acid method and hydroxylamine method , respectively .The protein expression of adiponectin receptor 1 ( AdipoR1) in the diaphragm was determined by immunohistochemistry and Western blot .RESULTS:The levels of blood lipids , blood glucose , FINS and HOMA-IR in HF and DM groups were higher than those in NC group , but decreased after L6H4 treatment.The serum APN level in HF and DM groups was lower than that in NC group , but increased after treat-ment with L6H4.The muscle fibers of the diaphragm were shrunk , fat particles accumulated in the muscle fibers , and the mitochondria were slightly swollen in HF and DM groups .The diaphragmatic fibrosis was obvious in DM group .These le-sions were relieved after L6H4 treatment.Compared with NC group, the level of MDA and the activity of SDH and NADH-TR in the diaphragm were increased in HF and DM groups , but decreased after treatment with L 6H4.The activity of SOD and the expression of AdipoR1 in the diaphragm were lower than those in NC group , but increased after L6H4 treatment. CONCLUSION: The curcumin analogue L6H4 exerts a protective effect on diaphragm in type 2 diabetic rats.The strengthened protein expression of AdipoR 1, the increased serum level of APN , and anti-lipid peroxidation may be involved in the process .
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Objective To compare the differences in setup error ( SE) assessment and correction between three-dimensional cone-beam computed tomography ( 3DCBCT ) and four-dimensional CBCT ( 4 DCBCT ) in breast irradiation patients during free breathing after breast-conserving surgery . Methods Twenty patients with breast cancer after breast-conserving surgery were recruited for external beam breast irradiation and 4DCBCT and 3DCBCT simulation. The target volumes were delineated. Volumetric modulated arc therapy plans were designed using the MONACO v510 treatment planning system. 3DCBCT and 4DCBCT images were collected alternately five times each before breast irradiation. The CT images were matched, and the interfraction SEs were acquired. After online setup correction, the residual errors were calculated, and the SEs, systematic errors, and random errors were compared. The paired t test was used for comparison between groups. Results The SEs acquired by 4DCBCT were significantly larger than those acquired by 3DCBCT in three directions ( P=0035, 0018, 0040 ) . After online setup correction, the random errors based on 3DCBCT were significantly smaller than those based on 4DCBCT in left-right and anterior-posterior ( AP ) directions ( 0.5± 039 mm vs. 0.7± 030 mm, P=0005;0.9± 109 mm vs. 1.2± 048 mm, P=0000) , and the residual errors based on 3DCBCT were also significantly smaller than those based on 4DCBCT in AP direction (0.2±033 mm vs. 0.6±063 mm, P=0000). The setup margins based on 4DCBCT was significantly larger than those based on 3DCBCT in AP direction both before and after online setup correction (P=0002). Conclusions Compared with 3DCBCT, 4DCBCT has more advantages in monitoring the SEs in three directions. Both 3DCBCT and 4DCBCT have similar efficacy in random error correction. The satisfying position repeatability and minimized target volume margins will be achieved by online setup correction.
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AIM:To investigate the protective effect of curcumin analogue L 6H4 on diaphragm of type 2 dia-betic rats.METHODS: SPF male Sprague-Dawley rats ( n=40) were randomly divided into 5 groups: normal control (NC) group, high fat (HF) group, high fat+L6H4 treatment (FT) group, diabetes mellitus (DM) group and DM +L6H4 treatment (DT) group.The rats in the later 4 groups were fed with high-fat diet.After 4 weeks of high-fat diet fee-ding, the rats in DM and DT groups were intraperitoneally injected with streptozotocin to induce type 2 diabetes melliutus. The rats in FT and DT groups were given L6H4 by gavage for 8 weeks.Blood glucose and blood lipid levels were detected biochemically .Fasting serum insulin ( FINS ) level was measured by radioimmunoassay and insulin resistance index (HOMA-IR) was calculated.Serum adiponectin (APN) level was measured by ELISA.The morphological changes of the diaphragm were observed under light and transmission electron microscopes .Lipid deposition and the activity of succinate dehydrogenase (SDH) and NADH-tetrazolium reductase (NADH-TR) were observed by enzyme histochemical staining . The content of malondialdehyde ( MDA) and the activity of superoxide dismutase ( SOD) in the diaphragm were measured by thiobarbituric acid method and hydroxylamine method , respectively .The protein expression of adiponectin receptor 1 ( AdipoR1) in the diaphragm was determined by immunohistochemistry and Western blot .RESULTS:The levels of blood lipids , blood glucose , FINS and HOMA-IR in HF and DM groups were higher than those in NC group , but decreased after L6H4 treatment.The serum APN level in HF and DM groups was lower than that in NC group , but increased after treat-ment with L6H4.The muscle fibers of the diaphragm were shrunk , fat particles accumulated in the muscle fibers , and the mitochondria were slightly swollen in HF and DM groups .The diaphragmatic fibrosis was obvious in DM group .These le-sions were relieved after L6H4 treatment.Compared with NC group, the level of MDA and the activity of SDH and NADH-TR in the diaphragm were increased in HF and DM groups , but decreased after treatment with L 6H4.The activity of SOD and the expression of AdipoR1 in the diaphragm were lower than those in NC group , but increased after L6H4 treatment. CONCLUSION: The curcumin analogue L6H4 exerts a protective effect on diaphragm in type 2 diabetic rats.The strengthened protein expression of AdipoR 1, the increased serum level of APN , and anti-lipid peroxidation may be involved in the process .
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Objective To compare the differences in setup error ( SE) assessment and correction between three-dimensional cone-beam computed tomography ( 3DCBCT ) and four-dimensional CBCT ( 4 DCBCT ) in breast irradiation patients during free breathing after breast-conserving surgery . Methods Twenty patients with breast cancer after breast-conserving surgery were recruited for external beam breast irradiation and 4DCBCT and 3DCBCT simulation. The target volumes were delineated. Volumetric modulated arc therapy plans were designed using the MONACO v510 treatment planning system. 3DCBCT and 4DCBCT images were collected alternately five times each before breast irradiation. The CT images were matched, and the interfraction SEs were acquired. After online setup correction, the residual errors were calculated, and the SEs, systematic errors, and random errors were compared. The paired t test was used for comparison between groups. Results The SEs acquired by 4DCBCT were significantly larger than those acquired by 3DCBCT in three directions ( P=0035, 0018, 0040 ) . After online setup correction, the random errors based on 3DCBCT were significantly smaller than those based on 4DCBCT in left-right and anterior-posterior ( AP ) directions ( 0.5± 039 mm vs. 0.7± 030 mm, P=0005;0.9± 109 mm vs. 1.2± 048 mm, P=0000) , and the residual errors based on 3DCBCT were also significantly smaller than those based on 4DCBCT in AP direction (0.2±033 mm vs. 0.6±063 mm, P=0000). The setup margins based on 4DCBCT was significantly larger than those based on 3DCBCT in AP direction both before and after online setup correction (P=0002). Conclusions Compared with 3DCBCT, 4DCBCT has more advantages in monitoring the SEs in three directions. Both 3DCBCT and 4DCBCT have similar efficacy in random error correction. The satisfying position repeatability and minimized target volume margins will be achieved by online setup correction.
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To further unravel adsorption mechanisms of effluent organic matter (EfOM) on the PVDF ultrafiltration membranes modified by nano-silica particles from micro perspective during different filtration phases, the membranes were prepared by adjusting the dosage of nano-silicon. The adsorption of EfOM on the surface of the membranes and the interaction between EfOM and the membranes were measured by quartz crystal microbalance with dissipation monitoring (QCM-D) and atomic force microscopy (AFM).The QCM-D results suggested that adsorbing capacity and adsorption rate of EfOM on the hydrophilic surfaces were lower than on the hydrophobic surfaces. Meanwhile, it was found that EfOM underwent adsorption via two steps: In the initial 15 min stage, a rapid adsorption of EfOM accumulated onto the membrane surface; The change in dissipation still occurred when the EfOM adsorption frequency reached balance, which demonstrated that the adsorption of EfOM remained unchanged on the membrane surfaces, and changes in the conformation of adsorption layer still occurred. For the AFM force test, it was found that the EfOM-membranes and EfOM-EfOM interactions declined with the increase of hydrophily, which revealed the essential reason for the decrease of adsorbing capacity and adsorption rate. The combined utilization of QCM-D and AFM effectively explained the effect of modified membranes on adsorption mechanisms of EfOM.
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To determine the fouling behavior of bovine serum albumin (BSA) on differet hydropniic PVDF ultrafiltration membrane over a range of pH, atomic force microscopy (AFM) and self-made colloidal probes were used to detect the microscopic adhesion forces of membrane-BSA and BSA-BSA, respectively. The results showed a positive correlation between the flux decline extent and the membrane-foulant adhesion force in the initial filtration stage, whereas the foulant-foulant interaction force was closely related to the membrane fouling in the later filtration stage. Moreover, the membrane-BSA adhesion interaction was stronger than the BSA-BSA adhesion interaction, which indicated that the fouling was mainly caused by the adhesion interaction between membrane and foulant. At the same pH, the adhesion force between PA membrane-BSA was smaller than that of PP membrane-BSA, illustrating the more hydrophilic the membrane was, the better the antifouling ability it had. The adhesion force between BSA-BSA fouled PA membrane was similar to that between BSA-BSA fouled PP membrane. These results confirmed that elimination of the membrane-BSA adhesion force is important to control the protein fouling of membranes.
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Incrustaciones Biológicas , Membranas Artificiales , Proteínas/química , Microscopía de Fuerza Atómica , Polivinilos , Albúmina Sérica Bovina/química , UltrafiltraciónRESUMEN
The hydrophilic modification of PVA composite membrane was applied in the reversed A2/O-MBR process to treat wastewater, the removal efficacy of COD, NH4(+) -N, TN, TP, turbidity and performance of composite membrane were investigated. The results indicate that the average removal rates of COD, NH4(+) -N and TP were higher than 90%, 95% and 80% under different reflux ratio, respectively. The reflux ratio had large impact on TN removal rate: when the reflux ratio was 100%, the removal rate was low; when the reflux ratio increases the range from 100% to 300%, the removal rate was correspondingly increased. Under the efficient interception of membrane, water turbidity was always less than 0.05NTU, and the composite film was controlled at (12 ± 0.5) L x (m2 x h)(-1) flux, the operation was uninterrupted for 52 days without any cleaning process of the membrane, the average rate of membrane fouling is 13.22 Pa x h(-1) and the process of membrane fouling was very slow. After FTIR analysis, we confirmed that polysaccharide and protein is a main composition of organic pollutants. LB is further proved to be the main pollutants from micro acting force between the membrane and the pollutants, which is consistent with FTIR analysis.
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Membranas Artificiales , Eliminación de Residuos Líquidos/métodos , Aguas Residuales/química , Compuestos de Amonio/análisis , Nitrógeno/análisis , Fósforo/análisis , Contaminantes Químicos del Agua/análisisRESUMEN
To further unravel the humic acid (HA) fouling mechanism during microfiltration under different conditions, such as pH, ionic strength, the concentration of calcium ions, atomic force microscopy (AFM) combined with self-made PVDF colloidal probe was applied to determine the relationship between the adhesion forces of membrane-HA or HA-HA and the flux decline of membrane. The results indicate adhesion forces were the main reason of membrane fouling. With the decrease of pH or increase of the ionic strength, due to the electrical neutralization caused by pH and electrical shielding effect of ionic strength, the adhesion forces of membrane-HA and HA-HA increased. Because of the comprehensive effect of "salt bridge" and electrical neutralization, there was a transition from increase to decrease for the adhesion forces of membrane-HA and HA-HA as the doses of calcium ions increased. In all cases, both of membrane-HA and HA-HA adhesion forces had the same variation tendency, which displayed a good correlation with the flux decline trends during fouling experiments, respectively, and provided certain theoretical support to further understand the formation mechanism of membrane fouling.
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Sustancias Húmicas , Membranas Artificiales , Ultrafiltración/métodos , Adhesividad , Calcio/química , Concentración de Iones de Hidrógeno , Iones , Microscopía de Fuerza AtómicaRESUMEN
Mixtures of polyvinylidene fluoride (PVDF) and polyvinyl alcohol (PVA) containing hydrophilic ultrafiltration membranes were prepared by adding PVA (5 to 30%) to PVDF by the phase inversion method. The hydrophilic contact angle (CA), equilibrium water content, pure water flux and bovine serum albumin retention were studied to assess the membrane performance. The anti-fouling performance of modified membrane to the secondary treated water was evaluated by flux decline, washing recovery rate and fouling resistance analysis. Scanning electron microscopy showed that the cross-section structure of the membranes had finger-like pores, which were well developed and uniformly distributed, and the sub-layer structure was looser and more porous with the increasing content of PVA. The CA gradually decreased. The steady flux was 800 L/m(2) h from P15 to P30, and the BSA retention sharply declined. The ultrafiltration tests for secondary treated water indicated that the main fouling source of the modified membrane was the concentration polarization and cake layer resistance. After physical flushing, the flux recovery ratio of the membrane could reach 100% when the PVA content was 5-15%, which shows excellent anti-pollution performance and good prospects for use in processing wastewater from urban sewage.
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Ciudades , Membranas Artificiales , Aguas del Alcantarillado , Ultrafiltración/instrumentación , Eliminación de Residuos Líquidos/instrumentación , Ultrafiltración/métodos , Eliminación de Residuos Líquidos/métodos , Purificación del AguaRESUMEN
Sow milk yield and quality is crucial for the survival and growth of piglets. To understand the molecular mechanisms of lactogenesis and lactation, mammary tissue samples were taken from six sows at -17(±2), 1 and 17(±2) days relative to parturition. Mammary tissues from two sows in the same stage were used to extract RNA, which were subsequently pooled in equal amounts. Nine pooled samples were hybridized to porcine Affymetrix GeneChips. Totally 1,524 genes were detected as significantly differentially expressed over the time course tested (p<0.01, q<0.01, fold change≥2 or ≤-2), including 709 upregulated and 575 downregulated genes identified at peak lactation compared to late pregnancy. Gene ontology analysis revealed that most of the upregulated genes were involved in transport, biosynthetic processes, and homeostasis, whereas most of the downregulated genes were involved in intracellular signaling cascades, cell cycle, and DNA replication. Furthermore, we identified 64 differentially expressed genes of the solute carrier families. Taken together, our microarray analysis provides insights into previously uncharacterized changes in transcriptome between late pregnancy and peak lactation in the porcine mammary gland. The solute carrier genes and other differentially expressed genes identified in this study will guide further characterization of their function to enhance milk yield and piglet growth.
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Lactancia/metabolismo , Glándulas Mamarias Animales/metabolismo , Animales , Femenino , Análisis de Secuencia por Matrices de Oligonucleótidos , Embarazo , Porcinos , Transcriptoma/genéticaRESUMEN
OBJECTIVE: To investigate the limited digestion of recombinant staphylococcal enterotoxin C2 (SEC2-His)in different conditions. METHODS: The purified recombinant SEC2-His was treated with different reagents and the cleavage of rSEC2 molecule was observed by SDS-PAGE. RESULT: The cleavage occurred in positions Cys93-Cys110 of the disulfide loop. Complete auto-cleavage of recombinant SEC2 was observed in solution at 37degrees within 24 hrs, and that was accelerated under alkaline conditions. The auto-cleavage of the recombinant protein was inhibited in the presence of beta-ME (2%), PMSF (5-10 mmol/L), imidazole (1 mol/L) or crude E.coli lysate. Non-specific degradation of recombinant SEC2 was promoted with the increasing of the concentration of H(2)O(2). CONCLUSION: The recombinant SEC2-His is broken down in special site of protein, which may be associated with the protein structure.
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Enterotoxinas/química , Proteínas Recombinantes de Fusión/química , Secuencia de Aminoácidos , Enterotoxinas/genética , Datos de Secuencia Molecular , Conformación Proteica , Estabilidad Proteica , Proteínas Recombinantes de Fusión/genéticaRESUMEN
<p><b>OBJECTIVE</b>To investigate the limited digestion of recombinant staphylococcal enterotoxin C2 (SEC2-His)in different conditions.</p><p><b>METHODS</b>The purified recombinant SEC2-His was treated with different reagents and the cleavage of rSEC2 molecule was observed by SDS-PAGE.</p><p><b>RESULT</b>The cleavage occurred in positions Cys93-Cys110 of the disulfide loop. Complete auto-cleavage of recombinant SEC2 was observed in solution at 37degrees within 24 hrs, and that was accelerated under alkaline conditions. The auto-cleavage of the recombinant protein was inhibited in the presence of beta-ME (2%), PMSF (5-10 mmol/L), imidazole (1 mol/L) or crude E.coli lysate. Non-specific degradation of recombinant SEC2 was promoted with the increasing of the concentration of H(2)O(2).</p><p><b>CONCLUSION</b>The recombinant SEC2-His is broken down in special site of protein, which may be associated with the protein structure.</p>
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Secuencia de Aminoácidos , Enterotoxinas , Química , Genética , Datos de Secuencia Molecular , Conformación Proteica , Estabilidad Proteica , Proteínas Recombinantes de Fusión , Química , GenéticaRESUMEN
Staphylococcal enterotoxins (SEs) are powerful superantigens that stimulate non-specific T-cell proliferation produced by Staphylococcus aureus and draw considerable attention as ideal drugs for cancer therapy. The filtrate of S. aureus culture has been used as ampul named Staphylococcal enterotoxin C injection in clinic for 10 years in China and proved to be effective. The superantigen SEC claimed to be the only active component without certifiable evidences. For further investigations of the active components of this injection and establishment of foundations for the development of novel anti-cancer drugs, in this research we extracted total DNA from S. aureus (FRI 1230), cloned, expressed and purified recombinant proteins of Staphylococcal enterotoxin M and N (rSEM and rSEN). The MTT assay of the purified rSEM and rSEN demonstrated that their abilities of stimulating T cells and inhibiting the proliferation of K562-ADM cells and B16 cells were equivalent to that of purified SEC2 in vitro. These findings suggested that SEC was not the only active component of Staphylococcal enterotoxin C injection and the effective procedure of expression and purification may be useful for mass productions of these therapeutically important proteins.
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Enterotoxinas/biosíntesis , Enterotoxinas/inmunología , Staphylococcus aureus/inmunología , Superantígenos/biosíntesis , Superantígenos/inmunología , Animales , Antígenos Bacterianos/biosíntesis , Antígenos Bacterianos/genética , Antígenos Bacterianos/inmunología , Clonación Molecular , Enterotoxinas/genética , Humanos , Ratones , Ratones Endogámicos ICR , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Staphylococcus aureus/genética , Superantígenos/genética , Linfocitos T/inmunología , Células Tumorales CultivadasRESUMEN
OBJECTIVE:To study the effect of moxifloxacin on pharmacokinetics of phenytoin sodium in rabbits. METHODS: A total of 10 rabbits were injected with phenytoin sodium (10 mg?kg-1) via the vein in ear edge,with serum samples taken at 10,30,60,120,240,360,and 480 min,respectively for establishing phenytoin sodium alone group. 48 h after the last sampling,the rabbits were given oral moxifloxacin (10 mg?kg-1) for 5 consecutive days;on day 6,the rabbits were injected with phenytoin sodium (10 mg?kg-1) at 2 h after oral administration of moxifloxacin,then serum samples were taken at different time points for establishing combination group of phenytoin sodium and moxifloxacin. The serum concentrations of phenytoin sodium at different time points were determined by UV-spectrophotometry and the pharmacokinetic parameters were computed with 3p97 program. RESULTS: The plasma concentration of phenytoin sodium reduced in the combination group compared with alone group. The AUC in the two groups was (5 836.22?489.13) vs. (4 329.21?344.67) mg?min?L-1 (P0.05). CONCLUSION: The elimination of phenytoin sodium was significantly accelerated by combination with moxifloxacin.
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AIM: To clone the gene of staphylococcal enterotoxin C2 and express it in the form of a soluble fusion protein in E. coli. Then the activation of SEC2 on mice lymphocyte and its lethal effects on tumor cells were studied. METHODS: Staphylococcus aureus SEC2 gene was cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEC2 was used to transform E. coli BL21, where the GST-SEC2 fusion protein was expressed efficiently. The rSEC2 protein was purified with Glutathione Sepharose 4B affinity column and digested with thrombin. The in vitro culture system was utilized to observe the activation of the SEC2 on mice lymphocyte and the lethal effects on tumor cells of the activated mice lymphocyte. RESULTS: The proper gene of SEC2 was cloned and purified rSEC2 was obtained. The MTT results indicated that rSEC2 have strong ability to stimulate mice lymphocyte to proliferate with a dose-dependent manner. With the proliferation of mice splenic lymphocyte, rSEC2 has a strong lethal effect on tumor cells B16, K562 and K562-AD. CONCLUSION: In this study, the gene of SEC2 was cloned and the rSEC2 protein was obtained, which had strong lethal effect on tumor cells B16, K562 and K562-AD.