Application and problems analysis of traditional Chinese medicine volatile oil preparation technology based on ionic liquids / 中国中药杂志

Ionic liquids(ILs) are salts composed entirely of anions and cations in a liquid state at or near room temperature, which have a variety of good physicochemical properties such as low volatility and high stability. This paper mainly reviewed the research overview of ILs in the application of traditional Chinese medicine(TCM) volatile oil preparation technology. Firstly, it briefly introduced the application of TCM volatile oil preparation technology and composition classification and physicochemical properties of ILs, and then summarized the application of ILs in the extraction, separation, analysis, and preparation of TCM volatile oil. Finally, the problems and challenges of ILs in the application of TCM volatile oil were explained, and the application of ILs in TCM volatile oil in the future was prospected.

Dosagem do magnésio eritrocitário: Qual é o seu real valor? / Measurement of erythrocyte magnesium: What is its real value?

O magnésio é um cátion de localização predominantemente intracelular e de grande importância em várias funções metabólicas. É possível que tenha uma participação importante em processos álgicos e inflamatórios. Sua dosagem sérica possivelmente não representa a realidade de sua concentração corporal. A mensuração do magnésio eritrocitário talvez possa representar um avanço na sua melhor avaliação

Magnesium is a cation with location predominantly intracellular and of great importance in several metabolic functions. It is possible that it plays an important role in pain and inflammatory processes. It's serum dosage possibly does not represent the reality of it's body concentration. The measurement of erythrocyte magnesium may represent an advance in its better evaluation.

A novel method for DNA delivery into bacteria using cationic copolymers

Amphiphilic copolymers have a wide variety of medical and biotechnological applications, including DNA transfection in eukaryotic cells. Still, no polymer-primed transfection of prokaryotic cells has been described. The reversible addition-fragmentation chain transfer (RAFT) polymer synthesis technique and the reversible deactivation radical polymerization variants allow the design of polymers with well-controlled molar mass, morphology, and hydrophilicity/hydrophobicity ratios. RAFT was used to synthesize two amphiphilic copolymers containing different ratios of the amphiphilic poly[2-(dimethyl-amino) ethyl methacrylate] and the hydrophobic poly [methyl methacrylate]. These copolymers bound to pUC-19 DNA and successfully transfected non-competent Escherichia coli DH5α, with transformation efficiency in the range of 103 colony-forming units per µg of plasmid DNA. These results demonstrate prokaryote transformation using polymers with controlled amphiphilic/hydrophobic ratios.

Proteomics and peptidomics analysis of Sepiae Endoconcha / 中国中药杂志

Shotgun based proteomics and peptidomics analysis were used to investigate the proteins and peptides in marine traditional Chinese medicine(TCM) Sepiae Endoconcha(cuttlebone). Peptides were extracted from cuttlebone by acidified methanol, and then strong cation exchange(SCX) resin was used to enrich those peptides. Also, proteins from cuttlebone were extracted and digested by trypsin. nano-LC Q Exactive Orbitrap mass spectrometry was used to analyze proteins and peptides from cuttlebone. As a result, a total of 16 proteins and 168 peptides were identified by protein database search, and 328 peptides were identified by De novo sequencing. The identified proteins were hemocyanin, enolase, myosin, actin, calmodulin, etc., and the identified peptides were derived from actin, histone, and tubulin. All these proteins and peptides were important components in cuttlebone, which would provide important theoretical and research basis for marine TCM cuttlebone investigations.

Effect of calcium on ion contents and expression of photosynthetic related genes in honeysuckle under salt stress / 中国中药杂志

Exogenous calcium can enhance the resistance of certain plants to abiotic stress. Research have demonstrated that exogenous calcium could enhances the resistance of honeysuckle under salt stress by promoting the transmission of photosynthetic electrons.The aim of this study was to investigate the effects of exogenous calcium on the contents of Na~+,K~+,Ca~(2+),Mg~(2+)and the expression of photosynthetic related genes Cab and rbc L. In this study,we used ICP-OES to analysis ion contents and used qRT-PCR to analysis the expression patterns of Cab and rbc L. The results showed that CaCl_2 significantly enhanced the K~+-Na~+,Ca~(2+)-Na~+,Mg~(2+)-Na+ratio of honeysuckle treated with 50 and 100 mmol·L~(-1) NaCl. Meanwhile,Cab and rbc L were significantly up-regulated under short-term salt stress,and CaCl_2 promoted this trend. From the two gene expression patterns,rbc L rapidly up-regulated on the first day of stress and then decreased,and was more sensitive to environmental changes. In summary,exogenous calcium could alleviate salt stress and increase plant development by increasing intracellular K~+-Na~+,Ca~(2+)-Na~+,Mg~(2+)-Na+ratio,and the transient overexpression of Cab and rbc L.

Tratamiento de agua contaminada con metales pesados utilizando como medios filtrantes biorresina intercambiadora de cationes de la cáscara y tallo de guineo y carbón activado de endocarpo de coco / Treatment of contaminated water with heavy metals, utilizing as filtering media a cation exchanger bio-resin from the banana peel and bush and activated carbon from coconut endocarp

En este trabajo de desarrollo experimental, la Escuela de Ingeniería Química de ITCA-FEPADE se propuso comprobar la efectividad de las cáscaras y pseudotallo de guineo y el endocarpo de coco, previamente tratados, para remover la contaminación por metales pesados en una muestra de agua. Para tal objeto, se procesaron dichas biomasas para ser utilizadas como medios filtrantes, los cuales se caracterizaron por medio de pruebas físicas: densidad y tamaño de partícula. Se evaluó su efectividad para remover metales, filtrando agua contaminada con cantidades conocidas de metales pesados tales como hierro, cromo y níquel (Fe3+Cr6+ y Ni2+), variando el tiempo de contacto y el tipo de medio filtrante. La cuantificación de los metales en el agua tratada se llevó a cabo por espectrofotometria de absorción atómica: para el níquel (λ = 232.0 nm); hierro (λ = 24830 nm) y cromo hexavalente (λ = 357.9 nm). Además, se determinó el color en los filtrados por el método de platino-cobalto. Se llegó a la conclusión que las biomasas utilizadas en este estudio resultaron efectivas para la disminución de metales pesados y color en la muestra de agua sintética elaborada en el laboratorio.

In this experimental development work, the Escuela de Ingeniería Química ITCA-FEPADE set out to verify the effectiveness of previously treated banana peels and pseudostem and endocarp to remove heavy metal contamination in a water sample. For this purpose, said biomasses were processed to be used as filter media, which were characterized by means of physical tests: density and particle size. Its effectiveness to remove metals was evaluated, filtering water contaminated with known amounts of heavy metals such as iron, chromium and nickel (Fe3 + Cr6 + and Ni2 +), varying the contact time and the type of filter medium. The quantification of the metals in the treated water was carried out by atomic absorption spectrophotometry: for nickel (λ = 232.0 nm); iron (λ = 24830 nm) and hexavalent chromium (λ = 357.9 nm). In addition, the color in the filtrates was determined by the platinum-cobalt method. It was concluded that the biomasses used in this study were effective in reducing heavy metals and color in the synthetic water sample prepared in the laboratory.

Elaboración de una biorresina intercambiadora de cationes a partir de cáscara de plátano o guineo para eliminar metales pesados en agua contaminada / Preparation of a cation exchanger bioresin start from banana peel or guineo to remove heavy metals in polluted water

Esta investigación tuvo por objetivo obtener una bio-resina intercambiadora de cationes utilizando cáscaras de guineo o plátano, la cual reduzca la concentración de metales pesados en agua contaminada. A esta bio-resina se le realizaron pruebas fisicoquímicas: densidad seca aparente, pH y solubilidad en agua y solventes orgánicos. Se evaluó su efectividad filtrando agua contaminada con metales pesados, tales como hierro, cromo y níquel (Fe3+, Cr6+ y Ni2+), variando las condiciones de tiempo de contacto, temperatura y el tipo de cáscara. La cuantificación de la concentración de los metales en el agua filtrada se llevó a cabo por espectrofotometría visible. Se llegó a la conclusión que la bio-resina obtenida es efectiva para disminuir la concentración de metales pesados en agua, teniendo especial afinidad química por el cromo hexavalente; metal pesado que logró remover arriba del 90%. Las condiciones óptimas de operación de la bio-resina son a 30°C y 90 minutos de tiempo de contacto con la muestra. Además, las pruebas fisicoquímicas, permitieron tipificarla preliminarmente como una resina de intercambio catiónico débil con un grado de entrecruzamiento bajo.

This research aimed to obtain a cation exchange bio-resin, using plantain or banana peels, that reduces the concentration of heavy metals in contaminated water. The bio-resin underwent physico-chemical tests like apparent dry density, pH, solubility in water and organic solvents. Its effectiveness was tested by filtering contaminated water with heavy metals such as iron, chromium and nickel (Fe3+, Cr6+ y Ni2+) and by varying contact time conditions, temperature and type of peel. The measure of concentration of metals in filtered water was taken through visible spectrophotometry. We concluded that the resulting bio-resin is effective in reducing the concentration of heavy metals in water, showing special chemical affinity with hexavalent chromium, a heavy metal that was able to remove above 90%. The bio-resin optimum working conditions are 30°C and 90 minutes of contact with the sample. Also, the physico-chemical tests allowed to preliminary typify it as a weak cation exchange resin with a low degree of crosslinking.

Elaboración de bio-resina intercambiadora de cationes a partir de cáscara de plátano o guineo para eliminar metales pesados en agua contaminada / Manufacture of cation exchange bio-resin from banana peel to remove heavy metals in contaminated water

Esta investigación tuvo por objetivo obtener una bio-resina intercambiadora de cationes utilizando cáscaras de guineo o plátano, la cual reduzca la concentración de metales pesados en agua contaminada. A esta bio-resina se le realizaron pruebas fisicoquímicas: densidad seca aparente, pH y solubilidad en agua y solventes orgánicos. Se evaluó su efectividad filtrando agua contaminada con metales pesados, tales como hierro, cromo y níquel (Fe3+, Cr6+ y Ni2+), variando las condiciones de tiempo de contacto, temperatura y el tipo de cáscara. La cuantificación de la concentración de los metales en el agua filtrada se llevó a cabo por espectrofotometría visible. Se llegó a la conclusión que la bio-resina obtenida es efectiva para disminuir la concentración de metales pesados en agua, teniendo especial afinidad química por el cromo hexavalente; metal pesado que logró remover arriba del 90%. Las condiciones óptimas de operación de la bio-resina son a 30°C y 90 minutos de tiempo de contacto con la muestra. Además, las pruebas fisicoquímicas, permitieron tipificarla preliminarmente como una resina de intercambio catiónico débil con un grado de entrecruzamiento bajo.

The objective of this research was to obtain a cation exchange bio-resin using banana peels, which reduces the concentration of heavy metals in contaminated water. Physicochemical tests were carried out on this bio-resin: apparent dry density, pH and solubility in water and organic solvents. Its effectiveness was evaluated by filtering water contaminated with heavy metals, such as iron, chromium and nickel (Fe3 +, Cr6 + and Ni2 +), varying the conditions of contact time, temperature and the type of shell. The quantification of the metal concentration in the filtered water was carried out by visible spectrophotometry. It was concluded that the bio-resin obtained is effective in reducing the concentration of heavy metals in water, having a special chemical affinity for hexavalent chromium; heavy metal that was able to remove over 90%. The optimal operating conditions for the bio-resin are at 30 ° C and 90 minutes of contact time with the sample. Furthermore, physicochemical tests allowed it to be preliminarily typified as a weak cation exchange resin with a low degree of crosslinking.

Variação espacial de cátions, ânions e variáveis físico-químicas no rio Solimões-Amazonas entre Manaus e Jutaí, bacia amazônica / Spatial variation of cations, anions and physicochemical variables in the Solimões-Amazonas River, between Manaus and Jutaí, Amazon Basin

Cátions e ânions majoritários são espécies químicas de grande importância nos diversos compartimentos aquáticos, porém, pouco abordados de forma exclusiva na bacia amazônica. Neste sentido, o objetivo deste estudo foi avaliar a dinâmica espacial de Na+, Ca2+, K+, Mg2+, Cl- e SO4 2- em um trecho do rio Solimões-Amazonas, levando-se em consideração a alcalinidade, potencial hidrogeniônico (pH), condutividade elétrica (CE), sólidos totais em suspensão (STS) e carbono orgânico total (COT). As coletas foram realizadas em maio, julho e setembro de 2012 em sete pontos na calha principal do rio Solimões-Amazonas e na foz de alguns dos principais tributários: rios Jutaí, Juruá, Japurá e Purus. Os cátions e ânions foram analisados por cromatografia de íons, STS por gravimetria, pH por potenciometria, alcalinidade por titulação potenciométrica, CE por condutimetria e o COT por oxidação catalítica por combustão. As concentrações médias dos cátions seguiram esta ordem Ca2+>Na+>Mg2+>K+ e dos ânions SO4 2->Cl-. De montante à jusante a tendência foi diminuição nas concentrações de cátions e ânions, das variáveis pH, CE, STS e alcalinidade, além do aumento das concentrações de COT. O presente estudo evidenciou mudanças nas características químicas sofridas pelo rio Solimões em seu curso. O aporte dos tributários pode ser apontado como um dos fatores responsáveis por essas modificações.

Major cations and anions are chemical species with great importance in different water compartments. However, no specific attention has been given to them in the Amazon basin. This study aimed to assess the spatial dynamic of Na+, Ca2+, K+, Mg2+, Cl- and SO4 2- in a specific section of the Solimões-Amazonas river taking into account the alkalinity, hydrogen potential (pH), conductivity electric (EC), total suspended solids (TSS) and total organic carbon (TOC). Samples were collected in May, July and September 2012 in seven points in the Solimões-Amazonas River and in the mouth of some of its main tributaries: rivers Jutaí, Juruá, Japurá and Purus. Cations and anions were analyzed by ion chromatography, TSS by gravimetry, pH by potentiometry, alkalinity by potentiometric titration, EC by conductimetry and TOC by catalytic oxidation combustion. The average cation concentrations followed this order: Ca2+ > Na+ > Mg2+ > K+ and anion SO4 2- > Cl. From the upstream of the river towards its downstream section there was a tendency for a decrease in cations and anions accumulation, pH, EC, TSS and alkalinity. On the other hand, there was an increase in TOC. The present study showed changes in chemical characteristics of Solimões River along its course. It was observed the contribution of the tributary rivers as one of the responsible factors for these changes.

Ectasia ductal mamaria en niños, a propósito de un caso clínico: A case report / Mammary duct ectasia in children

Introducción: La telorragia es un síntoma poco frecuente en pacientes pediátricos, la causa más frecuente en esta población es la ectasia ductal mamaria (EDM), que es una afección benigna y autolimitada, caracterizada por la dilatación del conducto mamario, fibrosis e inflamación periductal. Objetivo: Presentar un caso de EDM, para facilitar el rápido reconocimiento por parte de los médicos, y evitar estudios y tratamientos agresivos. Caso clínico: Lactante de sexo masculino de 6 meses de edad, sano, alimentado por lactancia materna exclusiva; consultó por un nódulo retroareolar derecho y telorragia unilateral. Se realizó una ecografía Doppler que mostró una lesión multiquística, sugerente de una EDM. Se planteó tratamiento expectante y acudió a control a los 6 meses con excelente evolución. Conclusiones: La EDM es la principal causa de telorragia en niños, corresponde a una afección benigna, y la resolución generalmente es espontánea, antes de los 9 meses. Por lo que su conocimiento es de gran relevancia para el adecuado diagnóstico y manejo de estos pacientes.

Introduction: Bloody nipple discharge is an infrequent symptom during childhood. The most common cause in this population is mammary duct ectasia (MDE), which is a benign and self-limiting condition, that is characterized by dilatation of the mammary ducts, fibrosis and periductal inflammation. Objective: Report of a case of MDE in order to improve physicians’ diagnosis accuracy and avoid aggressive studies and treatments. Case report: Six-months old male healthy infant, exclusively breastfeeded, that visited our clinic with a lump beneath his right nipple and bloody discharge from the same nipple. An ultrasound was performed which showed a multicystic lesion suggestive of MDE. Watchful waiting was decided as treatment, with good evolution after six months of follow up. Conclusions: The MDE is the leading cause of bloody discharge in pediatric population, being a benign condition that resolves spontaneously before nine months. The knowledge of this condition is essential so as to accurately diagnose and treat it.

Three-dimensional Culture of Chondrocyte Using Methacrylic Alginate Gel Beads Cross-linked with Mixed Metal-cation / 生物医学工程学杂志

This study was to explore a better three-dimensional (3-D) culture method of chondrocyte. The interpenetrating network (IPN) gel beads were developed through a photo-cross linking reaction with mixed barium ions and calcium ions at the ratio of 5:5 with the methacrylic alginate (MA), which was a chemically conjugated alginate with methacrylic groups. The second generation of primary cartilage cells was encapsulated in the MA gel beads for three weeks. In the designated timing, HE stain, Alamar blue method and Scanning electron microscopic were used to determine the cartilage cells growth, proliferation and the cell distribution in the scaffolds, respectively. The expression of type II collagen was investigated by an immunohistochemistry assay and the glycosaminoglycan content was quantitatively evaluated with the spectrophotometry of 1, 9 dimethylene blue assay. Compared to the alginate control group, the deposition of glycosaminoglycan was significantly upregulated in IPN-MA gel beads with higher cell proliferation. The secretion of extracellular matrix and proliferation of chondrocyte in methacrylic alginate gel beads were higher than that in Alginate beads. Cells were able to attach, to grow well on the scaffolds under scanning electron microscopy. The result of immunohistochemistry staining of collagen type II was positive, confirming the maintenance of chondrocyte phenotype in methacrylic alginate gel beads. This study shows a great potential for three-dimensional culture of cartilage.

Efficacy of RNA interference mediated by cationic liposomes / 生物工程学报

To investigate the cytotoxicity of the homemade peptide cationic liposome CDO14 and its efficacy of RNA interference (RNAi). MTT method was used to determine the cytotoxicity of the liposome to a human lung cancer cell line Luc-A549 that can express luciferase stably. Luciferase siRNA (Luc-siRNA) was transfected into Luc-A549 cells by CDO14. Contents of luciferase in the transfected cells were detected by luminous instrument and contents of total protein in these cells were detected by BCA method. Nude mice were inoculated with Luc-A549 cells in axilla to establish xenograft tumor model. Complexes of Luc-siRNA and the cationic liposomes were injected into the modeling mice via tail vein. Contents of luciferase in the transfected mice were detected by the whole body imaging system. The cytotoxicity of the homemade cationic liposome was similar to that of commercial liposome DOTAP, and lower than that of Lipo2000. The siRNA transfection efficacy mediated by CDO14 was higher than that mediated by DOTAP. The homemade peptide cationic liposome CDO14 is expected to serve as delivery vector in gene therapy because of its low cytotoxicity and high transfection efficiency.

Autocuidado do cuidador familiar de adultos ou idosos dependentes após a alta hospitalar / Self-care in family caregiver of dependent adults or elderly persons after hospital discharge / El autocuidado del cuidador familiar de adultos o ancianos dependientes tras el alta hospitalaria

Este artigo apresenta uma compreensão sobre o autocuidado do cuidador familiar segundo a teoria de Dorothea Orem. Resulta de uma pesquisa qualitativa com aporte da Teoria Fundamentada nos Dados, utilizando-se das técnicas de visita domiciliar, registro de notas de campo e entrevista semiestruturada com 11 cuidadores, após a internação de familiar em um Hospital Universitário de Minas Gerais. Foram obtidas quatro categorias, destacando uma categoria central, em torno da qual se analisaram as facilidades, dificuldades e estratégias para o autocuidado do cuidador. Entre as dificuldades, foram evidenciadas: tempo insuficiente para os cuidados com a saúde e, entre as facilidades, o apoio de outros familiares. As principais estratégias foram: apoio na fé; revezamento nos cuidados e recursos na comunidade. Concluiu-se que orientações no momento da alta e o acompanhamento de enfermagem após a alta contribuem para o autocuidado do cuidador, atuando sobre suas dificuldades e estimulando suas potencialidades.

This article presents an understanding concerning self-care in family caregivers according to Dorothea Orem's theory. Resulting from a qualitative research based on Grounded Theory, this work uses the techniques of home visiting, field notes and semistructured interviews with 11 caregivers after the hospitalization of a family member in a teaching Hospital located in Minas Gerais. Four categories were found and among them a central category is highlighted from which some facilities, difficulties and strategies for selfcare in caregiver were analyzed. Considering the difficulties, insufficient time for healthcare was noticed whereas the support from other family members appeared as a facility. The main strategies were: faith as a support; shift work in healthcare and community resources. This study demonstrated that hospital discharge guidelines and nursing follow-up after discharge were responsible for positive contributions to self-care in caregivers helping them to overcome their difficulties and enhancing their potentialities.

El artículo presenta una comprensión sobre el autocuidado del cuidador familiar, según la teoría de Dorothea Orem. Resulta de investigación cualitativa con aporte de la Teoría Fundamentada en los Datos, utilizándo se de las técnicas de visitas a domicilio; registro de apuntes de campo y entrevista semiestructurada, tras la internación de un familiar en un hospital universitario de Minas Gerais. Se llegó a cuatro categorías, señalando una categoría central, alrededor de la cual se analizaron las facilidades, dificultades y estrategias para el autocuidado del cuidador. Se evidenció, entre las dificultades, tiempo insuficiente para los cuidados con la salud y, entre las facilidades, el apoyo de otros familiares. Las principales estrategias fueron: apoyo en la fe, revezo en los cuidados y recursos en la comunidad. Se concluyó que orientaciones el momento del alta y el acompañamiento de enfermería tras el alta contribuyen para el autocuidado del cuidador, actuando sobre sus dificultades y estimulando sus potencialidades.

Cationic topical formulations tested in high dilutions offers protective effects in fibroblast cells culture

Topical administration is a simple and comfortable form of cutaneous administration of drugs. However, in this route of administration the drug needs to overcome the barrier posed by the skin to reach an effective concentration. For this reason, many topical formulations are developed with a cationic component. The promotion of absorption occurs due to the disruption of the stratum corneum. But this cationic component has also high irritating potential to the skin. The biotherapics are medicines prepared from a toxic product or etiologic agent, following the homeopathic pharmacopoeia technique, and they are used mainly in cases of hipersensitization. In this experiment, high dilutions (HD) obtained from a cationic formulation were prepared and evaluated considering cell viability in “in vitro” mouse fibroblast (L929) culture cells model by a colorimetric MTT assay. No signs of toxicity were observed, which demonstrates the safety of these HD preparations to the healthy cells. The effectiveness of these HD was also investigated in cells damaged by cationic formulations. The results demonstrated that the HD 30c was the most effective preparation in preventing the cell damage caused by the tested irritating product, increasing cell viability from 56.6% (damaged cells) to 100% (similar to negative control group, p>0.05). These results provide evidence of the positive action of high dilutions against the exposure to a cytotoxic agent.

Construction and characterization of liposomal magnetofection system in pig kidney cells / 生物工程学报

Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.

Influence of cationic cyclopeptide on microstructure and permeability of Caco-2 cell membrane / 药学学报

The microstructure of cationic cyclopeptide (TD-34) treated Caco-2 cell membrane was observed, and we discussed the relationship between membrane structure and insulin transmembrane permeability. Atomic force microscope (AFM) was used to observe living cell membrane in air condition and tapping mode. Results showed that the surface of Caco-2 cell membrane treated with TD-34 lost its smoothness and nearly doubled its roughness. Apparent permeability coefficients (P(app)) of insulin in Caco-2 cell monolayers increased 2.5 times. In conclusion, AFM can be used to observe microstructure of cationic cyclopeptide treated cell membrane and cationic cyclopeptide enhanced insulin delivery across Caco-2 cell membrane by increasing membrane fluidity.

Progress in the study of multidrug and toxin extrusion proteins / 药学学报

Mammal multidrug and toxin extrusion proteins (MATEs) play an important role in the transport of organic cations in the body. MATEs mediate the final excretion step for multiple organic cation drug used clinically and important endogenous substances. This article reviews the discovery, type, gene coding and polymorphism, body distribution, classification of substrates and inhibitors and their research method of MATEs. The article also discusses the major research significance of MATEs with examples.

Novel cationic liposome loading siRNA inhibits the expression of hepatitis B virus HBx gene / 药学学报

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.

Interactions between proteins and cation exchange adsorbents analyzed by NMR and hydrogen/deuterium exchange technique / 生物工程学报

In silico acquirement of the accurate residue details of protein on chromatographic media is a bottleneck in protein chromatography separation and purification. Here we developed a novel approach by coupling with H/D exchange and nuclear magnetic resonance to observe hen egg white lysozyme (HEWL) unfolding behavior adsorbed on cation exchange media (SP Sepharose FF). Analysis of 1D 1H-NMR shows that protein unfolding accelerated H/D exchange rate, leading to more loss of signal of amide hydrogen owing to exposure of residues and the more unfolding of protein. Analysis of two-dimensional hydrogen-hydrogen total correlation spectroscopy shows that lysozyme lost more signals and experienced great unfolding during its adsorption on media surface. However, for several distinct fragments, the protection degrees varied, the adsorbed lysozyme lost more signal intensity and was less protected at disorder structures (coil, bend, and turn), but was comparatively more protected against exchange at secondary structure domains (α-helix, β-sheet). Finally, the binding site was determined by electrostatic calculations using computer simulation methods in conjunction with hydrogen deuterium labeled protein and NMR. This study would help deeply understand the microscopic mechanism of protein chromatography and guide the purposely design of chromatographic process and media. Moreover, it also provide an effective tool to study the protein and biomaterials interaction in other applications.

An oxidant and organic solvent tolerant alkaline lipase by P. aeruginosa mutant: downstream processing and biochemical characterization

An extracellular alkaline lipase from Pseudomonas aeruginosa mutant has been purified to homogeneity using acetone precipitation followed by anion exchange and gel filtration chromatography and resulted in 27-fold purification with 19.6% final recovery. SDS-PAGE study suggested that the purified lipase has an apparent molecular mass of 67 kDa. The optimum temperature and pH for the purified lipase were 45°C and 8.0, respectively. The enzyme showed considerable stability in pH range of 7.0-11.0 and temperature range 35-55 °C. The metal ions Ca2+, Mg2+ and Na+ tend to increase the enzyme activity, whereas, Fe2+ and Mn2+ ions resulted in discreet decrease in the activity. Divalent cations Ca+2 and Mg+2 seemed to protect the enzyme against thermal denaturation at high temperatures and in presence of Ca+2 (5 mM) the optimum temperature shifted from 45°C to 55°C. The purified lipase displayed significant stability in the presence of several hydrophilic and hydrophobic organic solvents (25%, v/v) up to 168 h. The pure enzyme preparation exhibited significant stability and compatibility with oxidizing agents and commercial detergents as it retained 40-70% of its original activities. The values of Km and Vmax for p-nitrophenyl palmitate (p-NPP) under optimal conditions were determined to be 2.0 mg.mL-1 and 5000 μg.mL-1.min-1, respectively.