Polyphenol analysis in human milk by a rapid, cost-effective, and accurate micromethod: translational development / Análise de polifenóis no leite humano por meio de um micrométodo rápido, econômico e preciso: desenvolvimento translacional

ABSTRACT Objective: To develop a rapid method for analysing polyphenols, which are potentially active antioxidants against neonatal oxidative stress, from small human milk (HM) volumes. Methods: Acid and alkaline extractions were compared using two dyes: Folin-Ciocalteu and Fast Blue BB. Linearity, sensitivity, recovery percentage, polyphenol content, precision, and stability were assessed in 14 HM samples and compared using the Kruskal-Wallis H test (p<0.05). The best technique was applied to 284 HM samples to determine their polyphenolic content and its association with maternal diet by multifactorial linear regression. Results: Acidic extraction successfully recovered the gallic acid reference standard, whereas alkaline extraction overestimated it. Calibration curves for all methods were linear (R2>0.96) up to 500 mg/L. All bicarbonate-based Folin-Ciocalteu methods assayed were stable and repeatable, whereas Fast Blue BB-based variants were not. HM polyphenols (mean=94.68 mg/L) positively correlated to the dietary intake of hydroxycinnamic acids, the most consumed polyphenolic family in this population. Conclusions: A bicarbonate-based Folin-Ciocalteu micromethod allowed the accurate determination of polyphenols in HM, which might be useful for translational research settings and HM banks.

RESUMO Objetivo: Desenvolver um método rápido para analisar polifenóis, que são antioxidantes potencialmente ativos contra o estresse oxidativo neonatal, em pequenos volumes de leite humano (LH). Métodos: Foram comparadas extrações ácidas e alcalinas usando dois corantes: Folin-Ciocalteu e Fast Blue BB. Foram avaliadas variáveis como linearidade, sensibilidade, percentagem de recuperação, teor de polifenóis, precisão e estabilidade em 14 amostras de LH, comparadas usando o teste de Kruskal-Wallis H (p<0,05). A melhor técnica foi aplicada a 284 amostras de LH para determinar seu teor polifenólico e sua associação com a dieta materna por regressão linear multifatorial. Resultados: A extração ácida recuperou com sucesso o padrão de referência do ácido gálico, enquanto a extração alcalina o superestimou. As curvas de calibração para todos os métodos foram lineares (R2>0,96) até os 500 mg/L. Todos os métodos testados baseados em Folin-Ciocalteu com bicarbonato foram estáveis e repetíveis, enquanto as variantes baseadas em Fast Blue BB não. Os polifenóis do HM (média=94,68 mg/L) correlacionaram-se positivamente com a ingestão dietética de ácidos hidroxicinâmicos, a família de polifenóis mais consumida nesta população. Conclusões: Um micrométodo baseado em bicarbonato de Folin-Ciocalteu permitiu a determinação precisa de polifenóis no HM, o que pode ser útil para configurações de pesquisa translacional e bancos de HM.

Research progress on enteric-soluble soft capsule / 中国药房

Enteric-soluble soft capsule is a kind of new preparation that does not disintegrate in the stomach ，but releases rapidly in the intestinal tract to play a pharmacodynamic role. It has the unique advantages of improving drug stability ，reducing drug irritation ，delivering drugs directionally to the intestinal tract ，and prolonging drug action time. In this paper ，the decomposition and release mechanism ，application advantages ，classification of enteric-soluble coating materials and preparation methods of enteric-soluble soft capsule are sorted and summarized ，in order to provide reference for further development of this type of preparation.

Changes of cryoprecipitate coagulation factors prepared by different preparation methods and quick-freezing time / 中国输血杂志

【Objective】 To analyze the changes of fibrinogen (Fg) and Ⅷ factor levels of cryoprecipitated coagulation factors prepared by different methods and post-preparation quick-freezing time. 【Methods】 The fresh frozen plasma (FFP), prepared from 400mL whole blood, was randomly divided into 6 groups(group A1, A2, A3, B1, B2, B3) with 20 eliquots each, to prepare cryoprecipitate coagulation factors. Group A1, B1 were prepared by automatic cryoprecipitation preparation instrument. group A2, B2 applied the instrument after centrifugation and group A3, B3 were prepared manually. The quick-freezing time after preparation in group A1-A3 and B1-B3 were different(within 1 hour vs. more than 1 hour after preparation). The automated coagulation analyzer was used to measure Fg and Ⅷ factor levels in six groups, and further statistical analysis was carried out. 【Results】 The Fg content (mg) of six groups were 245.29±27.44 in group A1, 227.13±18.68 in group A2, 221.11±20.95 in group A3, 182.12±9.15 in group B1, 163.68±15.50 in group B2, and 155.61±19.28 in group B3, respectively. The Ⅷ factor levels(IU) were of six groups were 115.86±27.99 in group A1, 93.79±36.29 in group A2, 91.92±34.75 in group A3, 83.04±18.82 in group B1, 66.33±19.57 in group B2, and 69.34±13.26 in group B3, respectively. There were no significant differences in gender or age between group As and groups Bs. The levels of Fg and Ⅷ factors in group A1 were significantly higher than those in group A2 and group A3 (P<0.05). In addition, the levels of Fg and Ⅷ factors in group B1 were also obviously higher than those in group B2 and group B3 (P<0.05). Further, the levels of Fg and Ⅷ factors in group As were significantly higher than those in group Bs (P<0.05). 【Conclusion】 The automatic cryoprecipitation preparation instrument plus quick-freezing within 1 hour after preparation contribute to a higher efficiency and better quality than others.

Construction and application of gene delivery systems for primary dendritic cells / 中国药科大学学报

@#Nowadays, there is still no mature gene delivery system for safe and effective transfection on primary dendritic cells (DC). Herein, we constructed a liposome-based gene delivery system for primary DCs and optimized the preparation method to improve the transfection efficiency of siRNA on primary DCs. In this study, different methods, including co-incubation method, ethanol injection method, and protamine compound method, were used to prepare liposome/siRNA complexes based on different cationic lipids. Moreover, particle size, zeta potential, siRNA loading capacity, safety, stability, uptake efficiency and gene silencing efficiency of various liposome/siRNA complexes were detected to screen the optimal cationic lipid as well as its preparation method. We demonstrated that the OA2/siRNA delivery system prepared by the co-incubation method exhibited the best safety, uptake efficiency and gene silencing effect, compared to other siRNA delivery systems including the commercial Lipo2000. In summary, we provide a safe and effective gene delivery vector for primary DC cells through simple preparation method, which could also offer a gene delivery platform for other immune cells.

The preparation and evaluation of CRP trueness verificator: homogeneity, stability and commutability / 中华检验医学杂志

Objective To prepare the trueness verification materials of C-reactive protein (CRP) and evaluate its homogeneity, stability and commutability. Methods The high and low CRP concentrations trueness verification materials were from patient leftover sera which were pooled, mixed thoroughly, filtered and aliquoted. The homogeneity, stability and commutability of these materials were evaluated according to CNAS(China National Accreditation Service for Conformity Assessment, CNAS)-GL29:2010 "Reference materials-General and statistical principles for certification (ISO Guide35:2006)"and the Clinical and Laboratory Standards Institute (CLSI) EP30A. The trueness verification materials were used to evaluate the commutability in 10 clinical CRP detection systems, using forty-five patients' leftover sera with different CRP concentration evaluated by Deming regression in EP30A of CLSI. Meanwhile, the commutability of dilution series of ERM DA-474/IFCC were evaluated using the same method. Results A total of two CRP concentration level trueness verification materials were prepared, with high and low concentration levels of 754 and 743 vials, 1 ml each, respectively. The preparation showed good homogeneity (F<F0.05(14,30);On the condition of room temperature, 2-8 ℃ and -80 ℃, these materials were stable for 7 days and 44 months respectively, the slope of the linear equation of | b1 | less than t0.95,n-2 · s(b1), there was no statistically significant difference between the slope and zero, the stability is satisfied. The materials and the dilution series of ERM-DA 474/IFCC also showed good commutability among patient sera in 10 systems. Conclusions The trueness verification materials of C-reactive protein (CRP) showed good homogeneity, stability and commutability. The dilution series ERM DA-474/IFCC also have good commutability. These provided experimental support for the value transfer and application of the trueness verification materials .

Advancements in the preparation of high-performance liquid chromatographic organic polymer monoliths for the separation of small-molecule drugs / 药物分析学报

The various advantages of organic polymer monoliths, including relatively simple preparation processes, abundant monomer availability, and a wide application range of pH, have attracted the attention of chromatographers. Organic polymer monoliths prepared by traditional methods only have macropores and mesopores, and micropores of less than 50 nm are not commonly available. These typical monoliths are suitable for the separation of biological macromolecules such as proteins and nucleic acids, but their ability to separate small molecular compounds is poor. In recent years, researchers have successfully modified polymer monoliths to achieve uniform compact pore structures. In particular, microporous materials with pores of 50 nm or less that can provide a large enough surface area are the key to the separation of small molecules. In this review, preparation methods of polymer monoliths for high-per-formance liquid chromatography, including ultra-high cross-linking technology, post-surface modifica-tion, and the addition of nanomaterials, are discussed. Modified monolithic columns have been used successfully to separate small molecules with obvious improvements in column efficiency.

Limiar de positividade e sensibilidade dos métodos de Faust et al. e Lutz para detecção de cistos de Giardia duodenalis / Positivity threshold and sensitivity of methods of Faust et al. and Lutz for detection of Giardia duodenalis cysts

Objetivo: O presente estudo teve como objetivo determinar e comparar o limiar de positividade e a sensibilidade dos métodos de centrífugo-flutuação em sulfato de zinco (Faust et al.) e sedimentação espontânea (Lutz) para o diagnóstico de cistos de Giardia duodenalis. Métodos: Para obtenção de amostras fecais com quantidades conhecidas de cistos de G. duodenalis, amostras positivas para o parasito foram purificadas e quantificadas, e posteriormente alíquotas com diferentes quantidades foram adicionadas a amostras fecais negativas para parasitos. Após a contaminação de oito amostras negativas com quantidades variando entre 1.000 e 200.000 cistos por grama de fezes (c/g/f), as mesmas foram submetidas aos métodos de Faust et al. e Lutz, onde o primeiro se mostrou mais sensível para a detecção de cistos de G. duodenalis. Resultados: O limiar de positividade do método de Faust et al. foi de 11.000 c/g/f, e do método de Lutz foi de 100.000 c/g/f, portanto, cargas parasitárias inferiores a esses limiares levariam a resultados falso-negativos. Conclusão: O método de Lutz não é adequado para o diagnóstico de giardíase, portanto deve ser sempre utilizado em conjunto com o método de Faust et al.

Objective: The present study aimed to determine and compare the positivity threshold and sensitivity of the methods of zinc sulfate centrifugal flotation (Faust et al.) and spontaneous sedimentation (Lutz) for the diagnosis of Giardia duodenalis. Methods: To obtain fecal samples containing known amounts of G. duodenalis cysts, the samples with the parasite were purified and quantified, then aliquots with different amounts were added to fecal samples negative for parasites. After the contamination of eight negative samples with amounts ranging between 1.000 and 200.000 cysts per gram of feces, they were subjected to methods of Faust et al. and Lutz, where the first was more sensitive for the detection of G. duodenalis cysts. Results: The positivity threshold of the method of Faust et al. was 11.000 c/g/f, and the method of Lutz was 100.000 c/g/f, so parasitic loads below those thresholds would lead to false-negative results. Conclusion: The method of Lutz is not suitable for the diagnosis of giardiasis, therefore must always be associated with the method Faust et al.

Identificación molecular de genotipos papilomavirus humanos en pacientes con cáncer de cuello uterino / Molecular identification of human papilloma virus genotypes in patients with cervical cancer

Fundamento: la principal causa para el cáncer cervico uterino es el papilomavirus humano de alto riesgo. No existen antecedentes de estudios moleculares para la tipificación de papilomavirus humano en la población de Camagüey. La reacción en cadena de la polimerasa es una técnica de Biología Molecular que se ha usado desde siempre para el diagnóstico clínico; esta permite confirmar la presencia del ADN del Papilomavirus en el ADN total extraído a partir de muestras de pacientes con cáncer de cuello uterino. Objetivo: demostrar por primera vez los genotipos papilomavirus humano de alto riesgo circulantes, que causan cáncer de cuello uterino en la población femenina de Camagüey, Cuba. Métodos: se realizó un estudio analítico prospectivo donde se estudiaron 22 pacientes femeninas de la provincia de Camagüey, que fueron atendidas en la consulta de Patología de cuello del Hospital Ginecoobstétrico. La identificación y tipificación de los genotipos papilomavirus humano se realizó mediante el procedimiento molecular polimorfismo de longitud en los fragmentos de restricción. Resultados: el 63, 6 % de los pacientes presentaron lesiones tipo exofítica, el 4, 5 % endofítica y el 31, 8 % de otros tipos. Este estudio confirmó que los genotipos papilomavirus humanos de alto riesgo circulantes en la provincia Camagüey son los genotipos 16 y 31, donde el más frecuente fue el genotipo 16. Conclusiones: la presente investigación constituye el primer reporte de un estudio molecular de papilomavirus humanos a partir de muestras de pacientes con cáncer de cuello uterino en la provincia de Camagüey, Cuba. Estos resultados, junto a los obtenidos por otros autores, tienen una contribución importante en el diseño de preparados vacunales preventivos o terapéuticos, cada vez más efectivo hacia una solución anticipada para el cáncer de cuello uterino en Cuba.

Background: it is demonstrated that the main cause of cervical cancer is high risk humanp virus. There is no precedent of molecular studies for the typing of Human Papilloma Virus in the population of Camagüey. Polymerase chain reaction is Molecular Biology technique that has been used traditionally for the clinical diagnosis and other purposes. This technique allows confirming the presence of papillomavirus´DNA in the total extracted DNA, from samples of patients with cervical cancer. Objective: to demonstrate for the first time existing high-risk human papilloma virus genotypes that cause cervical cancer in female population of Camagüey, Cuba. Methods: a prospective analytic study was conducted, in which 22 female patients of the province of Camagüey were studied. They received medical attention at Ana Betancourt Hospital. Identification and typing of the Human Papilloma Virus genotypes was carried through the molecular procedure Restriction Fragment Length Polymorphism. Results: patients who presented exophytic lesions accounted for 63, 6%, 4, 5 % had endophytic type, and 31, 8 % presented other types. This study confirmed that high-risk Human Papilloma Virus genotypes existing in the province of Camagüey are genotypes 16 and 31, and the most frequent is 16. Conclusions: this research is the first report of a molecular study of Human Papilloma Virus from samples of patients with cervical cancer in the province of Camagüey, Cuba. These results, along with the ones obtained by other authors, make an important contribution in the design of the increasingly effective therapeutic and preventive vaccine to an anticipated solution to cervical cancer in Cuba.

Research progress of drug-loading gold nanoparticles / 药学实践杂志

In recent years ,as a novel drug delivery system ,gold nanoparticles have attracted widespread attention .Be-cause of their special physicochemical properties ,such as quantum size effect ,unique optical phenomenon ,easily reacting with thiol compounds or disulfides and so on ,gold nanoparticles can delivery variety of types of drugs ,like proteins ,nucleic acid , small molecular drugs ,therefore they can be applied in tumor treatment and detection .In this paper the preparation of drug-loading gold nanoparticles ,their drug-loading ways and safety issues were reviewed .

Preparation and Efficacy Evaluation of the Placebos of Sicao Tongmai Capsules / 中国药师

Objective:To prepare the placebos of Sicao Tongmai capsules to provide the reference for the placebo preparation in clinical trials of traditional Chinese medicines. Methods: Taking the placebo preparation of Sicao Tongmai capsules as the example, the appearance, color and smell of the placebo were assessed by the scoring method and an electronic nose. Results:The appearance, color and taste of the placebo were in line with those of the drug powder, while there was difference in smell. The simulated clinical ex-periments showed that the placebo was difficult to be unblinded. Conclusion:The placebo can be used in TCM clinical trials, howev-er, the production technology should be further improved and modified.