Progress in protein extraction methods for fungal identification with MALDI-TOF MS / 中华微生物学和免疫学杂志

With the development of techniques for rapid microbial identification, MALDI-TOF MS has become an important tool for clinical identification of fungi. Problems such as the applicability and standardization of protein extraction methods have hindered the development of MALDI-TOF MS technology in the fungal field. This paper analyzed the complex structure of fungal cell walls, introduced the protein extraction methods recommended by MALDI-TOF MS commercial mass spectrometry systems, discussed the protein extraction methods for the identification of various genera of yeast-like fungi and filamentous fungi by MALDI-TOF MS, such as direct smear method, formic acid acetonitrile extraction method and magnetic bead grinding method, and summarized the current status and drawbacks of protein extraction methods in fungal identification by MALDI-TOF MS with a view to providing theoretical reference for subsequent research.

Effects of different extraction methods on chemical compositions and biological activities of polysaccharides from Lycium barbarum / 中国中药杂志

In this study, five polysaccharides from Lycium barbarum(LBPs)(LBP-1-LBP-5) were selectively extracted by different extraction methods, and the chemical composition, structural characteristics, and biological activities of LBPs were explored. The results of chemical composition analysis showed that alkaloids were not detected in the five LBPs. The total polysaccharide content was(81.95%±1.6%)-(92.96%±0.76%), the uronic acid content was(8.26%±0.46%)-(24.81%±0.46%), and the protein content was(0.06%±0.03%)-(1.35%±0.13%). The monosaccharide compositions of the five LBPs were basically same, mainly including glucose, xylose, and galactose. However, there was significant difference in the content ratio of different monosaccharide. The results of infrared spectra analysis indicated that the five LBPs had typical infrared spectral characteristics of polysaccharides. The results of nuclear magnetic resonance characteristic spectrum analysis revealed that the five LBPs had two configurations of α and β. Meanwhile, there were triple helix structures in LBP-2, LBP-3, and LBP-4, which enhanced the activities of polysaccharides. The results of activities screening suggested that the biological activities of the five LBPs were significantly different. LBP-3 showed the highest lipid oxidation clearance rate, and its antioxidant activity was equivalent to that of the positive control group. The inhibitory rate of LBP-4 on α-amylase and its activation rate of alcohol dehydrogenase were better than those of other fractions, and the inhibitory rate of LBP-4 on α-amylase was slightly higher than that of the positive control group when the mass concentration was 10 g·L~(-1). LBP-2 showed stronger inhibitory activity against α-glucosidase and hyaluronidase. This study provides references for the precise development and utilization of LBPs.

Biological screening of herbal extracts and essential oil from Plectranthus species: α-amylase and 5-lipoxygenase inhibition and antioxidant and anti-Candida potentials

Abstract The phenolic compound content, the antioxidant and α-amylase inhibition potentials of different extracts of the Plectranthus amboinicus, P. barbatus and P. ornatus were evaluated. We also evaluated the influence of plant growth and harvest time on the chemical composition of the essential oil (EO) of P. amboinicus, its antioxidant and anti-Candida activities and the α-amylase and lipoxygenase inhibitions. The turbo-extract of P. barbatus showed the greatest phenolic compound content and antioxidant activity. No α-amylase inhibition activity was observed in the analyzed extracts, but the turbo-extraction and refluxing extracts possessed high antioxidant activities. Protected cultivation and morning harvest conditions gave the best antioxidant activities, which was associated to the highest carvacrol content. P. amboinicus EO antioxidant activity could contribute to the reduction of oxidative stress in diabetes. Causal Candida strains of diabetic foot ulcers showed sensitivity to P. amboinicus EO. C. albicans and C. dubliniensis were the most sensitive of the selected Candida strains. Turbo-extracts or refluxing of the three species extracts and the EO of P. amboinicus should be considered as a potential candidate for the management the complications of type 2 diabetes.

Composição de fenólicos, flavonoides, antocianinas, determinação da atividade antioxidante e comparação de métodos extrativos da folha e da casca de Caryocar brasiliense / Composition of Phenolic, Flavonoids, Anthocyanins, Determination of Antioxidant Activity and Comparison of Extractive Methods of Leaf and Bark of Caryocar brasiliense

Introdução: Caryocar brasiliense é conhecida popularmente como Pequi ou Pequizeiro, sendo uma planta com propriedades medicinais para tratamento de doenças respiratórias, úlceras gástricas e dores musculares. Objetivo: comparar a extração de biocompostos por meio de dois métodos extrativos, agitação magnética e banho ultrassônico. Além disso, avaliar a atividade antioxidante, o teor de compostos fenólicos, flavonoides e antocianinas em extratos das folhas e da casca de Pequi (C. brasiliense), com vistas a agregar valor quanto às suas propriedades funcionais. Metodologia: o conteúdo de compostos fenólicos foi determinado pelo método de Folin-Ciocalteu, flavonoides e antocianinas pelo método de Lima e Melo. A atividade antioxidante foi medida pelo método 2,2-difenil-1-picrilhidrazil (DPPH). Resultados: o extrato das folhas de Pequi exibiu maior teor de fenólicos em relação à casca, independente do método de extração. O extrato das folhas obtido em banho ultrassônico apresentou forte atividade antioxidante com valor de 72,2%. Conclusão: os extratos de Pequi demonstraram um perfil fitoquímico promissor que deve ser investigado no futuro para aplicação farmacológica como adjuvante ou precursor na síntese de novos cosméticos ou medicamentos com propriedades antioxidante.

Introduction: Caryocar brasiliense is a medicinal plant used in the treatment of respiratory diseases, gastric ulcers and muscle pain. Objective: to compare the extraction of natural compounds using two extractive methods, magnetic stirring and ultrasonic bath. In addition, to evaluate the antioxidant activity, the content of phenolic compounds, flavonoids and anthocyanins in the leaves and bark of Pequi (C. brasiliense), with a view to adding value through its functional properties. Methodology: the phenolic content was determined by the Folin-Ciocalteu method, flavonoids and anthocyanins by the Lima and Melo method. Antioxidant activity was measured using the 2.2-diphenyl-1-picrilhhydrazyl (DPPH) method. Results: the extract of the Pequi leaves exhibited a higher phenolic content in relation to the bark for both extraction methods. The Pequi leaf in an ultrasonic bath showed strong antioxidant activity with a value of 72.2%. Conclusion: Pequi extracts demonstrated a promising phytochemical profile that should be investigated in the future for pharmacological application as an adjuvant or precursor in the synthesis of a new cosmetic or medicine with antioxidant function.

Advances in isolation and extraction standardization and clinical translation of exosomes / 口腔疾病防治

@#Exosomes are phospholipid bilayer vesicles secreted by living cells that can carry a variety of signaling molecules, such as RNA, DNA, protein, and lipids. Exosomes play a role in the transmission of signaling molecules between cells, thus regulating many physiological and pathological processes. The methods of extracting exosomes include differential centrifugation, density gradient centrifugation, exclusion chromatography, ultrafiltration, coprecipitation, polymer immune affinity, microfluidic separation technology, etc. Each of these extraction technologies has advantages and disadvantages; however, there is no unified international standard. In addition, the expression of specific proteins and genetic material of exosomes from different cell sources are different; thus, their expression characteristics and functions are also distinctive. Based on this situation, research on exosomes is limited to preclinical studies, and difficulties and challenges still exist in clinical application. This paper summarizes the progress of research in the field of exosomes, to understand the characteristics, modification and application of exosomes from different cell sources, and to summarize their advantages and disadvantages as well as challenges, which can help researchers better understand and master the performance of exosomes. Furthermore, improvement of standard procedures in the extraction and manufacturing of exosomes is important, as it will provide a reference for researchers to carry out exosome-related translational clinical research.

A comparative study of the antifungal activity of essential oils of Varronia curassavica Jacq. obtained by different distillation methods / Estudo comparativo da atividade antifúngica dos óleos essenciais de Varronia curassavica Jacq. obtidos por diferentes métodos de destilação

This work aimed to compare the antifungal activity of the essential oil of Varronia curassavica obtained by hydrodistillation and microwave against the fungus Colletotrichum musae and verify the alterations caused by these extraction methods on the leaf surface. This study used four essential oil samples obtained by different methods, two by hydrodistillation [HD1 (1.0 L of water and 100 min.) and HD2 (2.0 L of water and 140 min.)] and two by microwave [MI1 (500W, 20 min, without water) and MI2 (700W, 40 min, with 50 mL of water added to fresh leaves)]. Essential oils concentrations of 0.05, 0.1, 0.5, 1.0, and 3.0% (v / v) were tested in PDA medium. The mycelial growth of C. musae was evaluated by measuring the diameter, every 24 hours up to 144 hours after the beginning of the incubation. Untreated leaves and leaves treated with HD1 and MI1 were prepared for observation in a scanning electron microscope (SEM) LEO EVO 40. The most abundant compounds detected in the essential oil samples analyzed by gas chromatography were: shyobunol, germacrene D-4-ol, E-caryophyllene, bicyclogermacrene, and α-cadinol. Up to 72 hours after the beginning of the incubation, C. musae presented no mycelial growth, even at the lowest essential oil concentration. Conversely, mycelial growth was detected in the control (PDA + DMSO) from 24 hours after incubation. At 144 hours after incubation, regardless of the concentration, the essential oil samples obtained by HD provided lower mycelial growth of C. musae (1.49 cm) when compared with samples obtained by MI (1.80 cm). This difference possibly occurred due to the reduction to less than half of the germacrene D-4-ol content in the samples obtained by MI. The four essential oil samples tested inhibited the mycelial growth and thus presented a inhibitory effect on C. musae. The SEM revealed more drastic changes on the surface of the leaf treated with MI than on those treated with HD. The essential oil of V. curassavica, mainly when obtained by hydrodistillation, has the potential for use in the control of C. musae.

O objetivo do trabalho foi comparar a atividade antifúngica do óleo essencial de Varronia curassavica obtido por hidrodestilação e micro-ondas frente ao fungo Colletotrichum musae e verificar as alterações que esses métodos de extração causam na superfície da folha. Quatro amostras de óleo essencial obtidas em diferentes condições foram utilizadas. Sendo duas por hidrodestilação, HD1 (1,0 L de água e 100 min.) e HD2 (2,0 L de água e 140 min.); e duas por micro-ondas, MI1 (500W, 20 min. sem adição de água) e MI2 (700W, 40 min. com adição de 50 mL de água às folhas frescas). Foram testadas as concentrações 0,05; 0,1; 0,5; 1,0 e 3,0 % (v/v) de óleo essencial em meio BDA. O crescimento micelial do C. musae foi avaliado por medições do diâmetro, a cada 24 horas até 144 horas após o início da incubação. Folhas sem qualquer tratamento e após os tratamentos HD1 e MI1 foram preparadas para observação em microscópio eletrônico de varredura (MEV) LEO EVO 40. Os compostos mais abundantes nas amostras de óleo essencial analisadas por cromatografia gasosa foram: shyobunol, germacreno D-4-ol, E-cariofileno, biciclogermacreno e α-cadinol. Até 72 horas após o início da incubação, não foi observado nenhum crescimento micelial do C. musae, mesmo nas concentrações mais baixas de óleo essencial, enquanto, para o controle (BDA + DMSO), foi observado crescimento do fungo a partir de 24 horas. Após 144 horas, independentemente da concentração, as amostras de óleo essencial obtidas por HD proporcionaram menor crescimento micelial do C. musae (1,49 cm) quando comparadas às amostras obtidas por MI (1,80 cm). Possivelmente essa diferença ocorreu devido à redução para menos da metade, do teor de germacreno D-4-ol, nas amostras obtidas por MI. As quatro amostras de óleo essencial testadas foram capazes de inibir o crescimento micelial, apresentando portanto, um efeito inibitório sobre o C. musae. Alterações mais drásticas observadas através da MEV foram visualizadas na superfície da folha submetida ao processo de extração por MI em comparação à HD. O óleo essencial de V. curassavica, sobretudo o obtido por hidrodestilação, apresenta potencial para o controle de C. musae.

Tratamiento del cuerpo extraño rectal en un Servicio de Urgencia / Rectal foreign body treatment at an urgency service

Resumen Introducción: El cuerpo extraño rectal (CER) es un problema de difícil manejo para el cirujano general. La vía anal es la puerta de entrada más habitual seguido de la oral. La sospecha clínica es fundamental para el diagnóstico, pudiendo apoyarse de imágenes. El tratamiento es su extracción por vía transanal o por vía anterior. Objetivos: Caracterizar, describir y presentar el tratamiento de los pacientes con CER que consultaron en urgencia del Hospital el Pino (HEP) entre los años 2011 y 2016. Material y Método: Estudio serie de casos. Se realiza revisión de fichas clínicas de pacientes con CER en un periodo de 5 años en el Servicio de Urgencia del HEP. Se identificó a los pacientes tratados en box y a aquellos tratados en pabellón. Se caracterizó según variables demográficas, tratamiento, complicaciones y días de hospitalización. Resultados: Se identificaron 18 pacientes, 13 hombres y 5 mujeres de 45 años (± 36-51) y 34 años (± 23-64) respectivamente. En 1 de ellos fue posible extracción manual transanal en box sin anestesia y en 17 se requirió tratamiento en pabellón, donde 12 fue por vía transanal y 5 por abordaje abdominal. En ningún caso se identificó lesión intestinal. Evolución postoperatoria adecuada. Días de hospitalización promedio 1,8 días. Conclusión: Este es un problema infrecuente y de difícil manejo. Existe mayor frecuencia en hombres de edades media siendo la puerta de entrada la vía anal en todos los casos. En la mayoría se requirió extracción en pabellón bajo anestesia siendo el abordaje transanal el más utilizado y con buenos resultados.

Introduction: The rectal forcing body (RFB) is a problem which involves a difficult management for surgeons. Anal via is the most frequent entrance area, followed by the oral via. Clinical suspicion is fundamental for diagnosis and it is possible to support it using imaging. Treatment considers extraction through transanal or frontal via. Aim: To describe and present the treatment of patients with rectal foreing body in the Urgency Service of El Pino Hospital during years 2011 to 2016. Materials and Method: Case studies. Clinical records revision of patients with rectal foreing body in the Urgency Service of El Pino Hospital considering a 5-year period. Patients are identified as those treated in the box and in a surgical pavilion. Patients were classified according to demographic variables, received treatment, associated complications and hospitalization time. Results: 18 patients are identified, 13 men and 5 women, 45 (± 36-51) and 34 (± 23-64) years respectively. It was possible to manually extract via trans-anal at box, in one of these cases; 17 of them required treatment at surgical pavilion, where 12 were via trans-anal and 5 via abdominal. None of these cases had intestinal injury. Post-surgery evolution was adequate. Average hospitalization was 1.8 days. Conclusion: RFB is a non-frequent problem which requires high diagnose suspicious to be able to carry out proper treatments. It is more frequent in middle age men, with transanal input. In most cases, the extraction was done in surgical pavilion and required anesthesia and were done via trans anal with good results.

Paying attentions to advantages and disadvantages of intraocular femtosecond laser in cataract surgery / 中华实验眼科杂志

Requirements for cataract surgery have gradually increased from the original rehabilitation procedure to personalized refraction surgery.Femtosecond laser-assisted cataract surgery is being widely used because of its precision of incision,accuracy of capsule cutting and high efficiency of nuclear fracturing in recent year.Currently some operating systems of femtosecond laser-assisted cataract surgery include LenSx,Catalys,LenSAR and Victus,and they are different in the design of contact interfaces,video and operating bed and therefore present different working methods.Ophthalmologists should be aware of the differences of these operating systems and accurate assess the clinical findings of patients in order to consider the preciseness,accuracy,efficiency of incision,capsulorhexis and nucleus fragmentation to avoid the intraoperative and postoperative complications.

Chemical constituents and pharmacological effect of eaglewoood: research advances / 国际药学研究杂志

Eaglewood is a rare and precious traditional medicine in China. It mainly contains the characteristic sesquiterpenes,2-(2-phenylethyl)chromones,aromatic compounds and the others. Eaglewood is used to treat disorders related to the digestive system,respiratory system,cardiovascular system,and central nervous system. This review sum- marizes the chemical constitutuents and pharmacological effect of eaglewood,so as to provide a reference for further re- search and development of eaglewood.

Sustainable production with obtaining glucosamine from crab exoskeletons / Produção sustentável para obtenção de glucosamina a partir de exoesqueletos de caranguejo

ABSTRACT: Two chemical treatments, five enzymatic (pectinase, lipase, hemicellulase, hemicellulose-cellulase or lipase-pectinase) and one microbiological (Bacillus subtilis) treatment were evaluated to obtain glucosamine hydrochloride (Gluc-HCl) from the chitin obtained from crab (Callinectes bellicosus) exoskeletons. Chemical treatments were referred as Method A (HCl hydrolysis during 75 min at 90°C) and Method B (HCl hydrolysis during 20 min and 14 h of rest). Glucosamine and, in some cases, N-acetyl-D-glucosamine were identified and quantified by HPLC. Treatments with the greater concentrations of Gluc-HCl in descending order were: lipase (94.4 mg/g), microbiological (45.7 mg/g), lipase-pectinase (22.9 mg/g), hemicellulase-cellulase (20.9 mg/g), hemicellulase (15.3 mg/g), pectinase (10.7 mg/g), Chemical A (7.3mg/g) and Chemical B (7.3mg/g). In terms of yield, the best treatments in descending order were: pectinase (94%), microbiological (94%), hemicellulase (92%), lipase (91%), Chemical B (88%), lipase-pectinase (88%), hemicellulase-cellulase (86%) and Chemical A (28.5%). The two most profound treatments were lipase and microbiological, so they are proposed as part of a viable method to produce Gluc-HCl from crab exoskeletons; they are ecofriendly procedures and could add value to the crab´s productive chain.

RESUMO: Dois tratamentos químicos, cinco enzimáticos (pectinase, lipase, hemicelulase, hemicelulose-celulase ou lipase-pectinase) e um microbiológico (Bacillus subtilis) foram avaliados para obter o cloridrato de glucosamina (Gluc-HCl) da quitina obtida a partir de exoesqueletos de caranguejo (Callinectes. Bellicosus). Os dois tratamentos químicos foram nomeados como método A (hidrólise de HCl para 75 min a 90 °C) e método B (hidrólise de HCl para 20 min e 14 h de repouso). A Glucosamina e, em alguns casos, N-acetil-D-glucosamina foram identificados e quantificados por HPLC. Os tratamentos em que as melhores concentrações de Glucosamina-HCl foram obtidas, em ordem decrescente: lipase (94,4 mg/g), microbiológica (45,7 mg/g), lipase-pectinase (22,9 mg/g), hemicelulase-celulase (20,9 mg/g), hemicelulase (15,3 mg/g), pectinase (10,7mg/g), Quïmica A (7,3 mg/g) e Quïmica B (7,3 mg/g). Em termos de produtividade, os melhores tratamentos em ordem decrescente foram: pectinase (94%), microbiológica (94%), hemicelulase (92%), lipase (91%), química B (88%), lipase-pectinase (88%), hemicelulase- celulase (86%) e produto químico A (28,5%). Os dois melhores tratamentos foram lipase e microbiológicos, propostos como método viável para obtenção de Gluc-HCl a partir de exoesqueletos de caranguejo; cumprem procedimentos ecologicamente corretos e podem agregar valor à cadeia produtiva do caranguejo.

Seasonal effect in essential oil composition and antioxidant activity of Plectranthus amboinicus leaves / Efeito sazonal na composição e atividade antioxidante do óleo essencial de malvarisco (Plectranthus amboinicus)

In view of the increasing search for natural products with biological activities, such as essential oils are of high quality because of their therapeutic and economic importance and represent the second class of natural compounds with the largest number of active constituents and production through Plants minimizes the environment and sanitary impacts . The objective of the present work is to assess the seasonal effect on the composition and antioxidant activity of essential oil in leaves of Plectranthus amboinicus (Lour.) Spreng using hydro and steam distillation techniques. With regarding to the extraction methodology of essential oils from the P. amboinicus, the hydrodistillation technique is the most efficient in obtaining the volatile product with the best features for the determination of antioxidant activity. The analyses of the essential oil indicated a similarity in their composition, being observed the Carvacrol as major compound in almost all analysis, except in January 2015 using steam distillation extraction. Regarding antioxidant activity and seasonality, it was verified that in October 2014 obtained the best value of inhibitory concentration, with IC50 = 124.97 ppm. However, it was possible to obtain the essential oil from the P. amboinicus leaves through hydro and steam distillation methodologies, but also noted the seasonality influence on it's the context of antioxidant capacity.

Tendo em vista a crescente busca de produtos naturais com atividades biológicas, os óleos essenciais apresentam alta qualidade devido à sua importância terapêutica e econômica e representam a segunda classe de compostos naturais com o maior número de constituintes ativos com produção através das plantas, o que minimiza os impactos ao meio ambiente. O objetivo do presente trabalho foi avaliar o efeito sazonal na composição e na atividade antioxidante do óleo essencial presente nas folhas da Plectranthus amboinicus (Lour.) Spreng utilizando as técnicas de hidrodestilação e arraste a vapor d'água. No que se refere à metodologia de extração dos óleos essenciais do malvarisco, constatou-se que a técnica de hidrodestilação é a mais eficiente na obtenção do produto volátil com as melhores características para a determinação da atividade antioxidante. Quanto à sazonalidade, verificou-se que no mês de Outubro/2014 obteve-se o melhor valor da concentração inibitória com CI50=124,97 ppm. Este trabalho mostra que é a obtenção do óleo essencial das folhas do malvarisco, utilizando as metodologias de hidrodestilação e arraste a vapor de água, como também, constatar a influência da sazonalidade na determinação da capacidade antioxidante do mesmo.

A randomized controlled clinical study on reverse-chopper prechop technique with phaco-and-chop technique for high myopia associated with hard nucleus cataract / 中华实验眼科杂志

Background The innovation of pre-chop technique lies in shortening surgery duration,reducing ultrasound power,and deseasing the loss of corneal endothelial cells (CECs).Objective This study was to compare the efficacy and safety of pre-chop technique using a reverse-chopper and phaco-and-chop phaco technique to super high myopia associated with hard nucleus cataract.Methods A prospective randomized-controlled-clinical interventional study was performed.Fourty eyes of 40 high myopia associated with cataract patients with Ⅲ-ⅣV degree of nucleus were enrolled in Beijing Tongren Hospital from March to September 2016.The patients were randomized into the pre-chop group and matched phaco-and-chop group according to random number table,and the self-made reverse chopper-assisted pre-chop phacoemulsification (phaco) surgery and phaco-and-chop phaco surgery were performed on the eyes of different groups,respectively.The phaco power,effective phaco duration and best corrected visual acuity (BCVA) were recorded and compared between the two groups to evaluate the efficacy,and the loss rate of corneal endothelial cells and eyes in different grades of cornea edema after operation were compared between the two groups to assess the safety of surgery procedure.Written informed consent from each patient was obtained prior to relevant examination and surgery.Results The surgery was finished smoothly in the eyes.The mean effective phaco duration was (47.30±11.29) seconds and (57.70± 14.51) seconds in the pre-chop group and phaco-andchop group,respectively,with a significant difference between them (t =-2.530,P =0.016).The BCVA was better at the seventh day after surgery in the pre-ehop group than that in the phaco-and-chop group (4.75t0.11 vs.4.67± 0.14),showing a significant difference between the two groups (t=2.147,P=0.038).In 1 month after surgery,the CECs loss rate was (10.82±3.77)％ in the pre-chop group,which was lower than (16.11±6.47)％ in the phacoand-chop group (t=-3.758,P=0.001).The number of the eyes with grade 2-3 of corneal edema in the pre-chop group was significantly decreased in comparison with the phaco-and-chop group 7 days after surgery (Z =11.822,P=0.008).Conclusions Compared with the conventional phaco-and-chop technique,reverse-chopper prechop technique appears to have a better efficacy and safety in phaco surgery for high-myopia associated with hard nuclear cataractous eyes.

Determination of betulinic acid, oleanolic acid and ursolic acid from Achyranthes aspera L. using RP-UFLC-DAD analysis and evaluation of various parameters for their optimum yield.

Achyranthes aspera L. is a well known herb commonly used in traditional system of Indian medicine to treat various disorders, such as cough, dysentery, gonorrhea, piles, kidney stone, pneumonia, renal dropsy, skin eruptions, snake bite, etc. Here, we used RP-UFLC-DAD method for determining triterpenoids betulinic acid (BA), oleanolic acid (OA) and ursolic acid (UA) from A. aspera. Optimum yield of these compounds were studied and evaluated using parameters viz., method of extraction, time of extraction, age of plant and plant parts (leaves, stem and roots). Linear relationships in RP-UFLC-DAD analysis were obtained in the range 0.05-100 µg/mL with 0.035, 0.042 and 0.033 µg/mL LOD for BA, OA and UA, respectively. Of the variables tested, extraction method and parts used significantly affected content yield. Continuous shaking extraction (CSE) at ambient temperature gave better extraction efficiency than exposure to ultra sonic extraction (USE) or microwave assisted extraction (MAE) methods. The highest content of BA, OA and UA were determined individually in leaf, stem and root extracts with CSE. Collective yield of these triterpenoids were higher in leaf part exposed to 15 min USE method. To best of our knowledge, the study newly reports UA from A. aspera and the same was confirmed using ATR-FT-IR studies. This study explains the distribution pattern of these major triterpenoids and optimum extraction parameters in detail.

A comparative study of cystotome-assisted prechop technique with stop-and-chop technique for pseudoexfoliation syndrome with cataract / 中华实验眼科杂志

Background Pseudoexfoliation syndrome (PEX) has a high incidence in Uygur population and usually leads to secondary glaucoma and complicated cataract.The abnormal change of lens tissue and degeneration of zonular fibers bring a lot of difficulties for phacoemulsification (phaco) with intraocular lens implantation,especially stop-and-chop phaco technique.Prechop technique is a new choping technology,but its application in PEX with cataract is less.Objective This study was to compare the efficacy and safety of pre chop phaco technique and stop-and-chop phaco technique for PEX combined cataract.Methods A randomized controlled Clinical trial was designed.Forty-one eyes of 41 patients with PEX combined cataract of Ⅲ degree of nucleus were enrolled in People's Hospital of Hetian District from March 2015 to January 2016.The patients were randomized into the prechop group and stop-and-chop group according to random nubmer table,and cystotome-assisted prechop phaco surgery and stop-and-chop phaco surgery were performed in different groups,respectively.The effective phaco duration,corneal endothelium loss rate,cornea edema eye number after operation,vision outcomes and complications were compared between the two groups.Results The mean effective phaco duration was 14.0 (13.0,16.5) minutes and 18.5 (16.5,24.0) minutes in the prechop group and stop-and-chop group,with a significant difference between them (Z =17.354,P ＜ 0.01).The corneal endothelial cells were (2 101.90 ± 209.08)/mm in the prechop group,and the number was similar to (2 002.30 ± 207.04)/mm of the stop-and-chop group (t =-1.530,P =0.134).Corneal endothelial cell lossing rate was (8.27±2.23)％ in the prechop group,which was lower than (13.09±4.26)％ in the stop-and-chop group (t =3.810,P =0.001).The BCVA was better in the prechop group than that in the stop-and-chop group in postoperative day 3 (P =0.044),and the corneal edema degree was not signigicantly different in postoperative day 1 and day 3 between the two groups (P=0.221,0.446).Intraoperative complication was rapture of zonule and occurred in 1 eye and 2 eyes in the prechop group and stop-and-chop group,respectively.Conclusions Compared with the stop-and-chop phaco technique,the prechop phaco tequnique can decrease intraoperative complication,lighten the postoperative damage of corneal endothelial cells and accelerate visual rehabilitation in PEX combined with cataract patients.

Enchytraeid abundance in Araucaria Mixed Forest determined by cold and hot wet extraction

Abstract Enchytraeids are small oligochaetes found worldwide in soils with sufficient moisture and organic matter, but scarcely studied in the Southern hemisphere. This is the third study on enchytraeid abundance in Brazil using wet extraction and the first carried out in Araucaria Mixed Forest (subtropical region). The sampling and extraction were based on the standard method ISO 23611-3/2007 using an adapted split soil corer and wet extraction with and without heat to assess the abundance of enchytraeids in a forest fragment at Embrapa Forestry in Colombo, Paraná State. The samplings were performed in 3 occasions between September 2011 and April 2012. The average numbers estimated by each method varied from appr. 2.000-12.000 (cold) and 5.000-12.000 ind./ m2 (hot), respectively, with a maximum of 44.000 ind./ m2 in one of the samples, the highest value reported so far in Brazil. The hot extraction was more advantageous, given the speed and preservation of the specimens in vivo, allowing taxonomic identification. Advantages and disadvantages of wet extractions compared to handsorting and formol methods are also discussed. Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia and Achaeta were the genera identified in the samples.

Resumo Os enquitreídeos são pequenos oligoquetas encontrados no mundo todo em solos com suficiente umidade e matéria orgânica, porém muito pouco estudados no hemisfério Sul. Este é o terceiro estudo sobre a abundância de enquitreídeos no Brasil utilizando o método de extração úmida e o primeiro realizado em Floresta Ombrófila Mista (região subtropical). A amostragem e extração foram baseadas no método padrão ISO 23611-3/2007, utilizando-se um trado desmontável adaptado e extração úmida com e sem aquecimento para acessar a abundância de enquitreídeo em um fragmento de floresta na Embrapa Florestas em Colombo, Paraná. As amostragens foram realizadas em três ocasiões entre setembro, 2011 e abril 2012. Os números médios estimados através de cada método variaram de 2.000-12.000 (frio) e 5.000-12.000 ind./ m2 (quente), respectivamente, e o máximo de 44.000 ind./ m2 em uma das amostras, o mais alto já relatado no Brasil. A extração quente foi a mais vantajosa, considerando a rapidez e preservação dos exemplares in vivo. As vantagens e desvantagens das extrações úmidas comparadas aos métodos de triagem manual e extração com formol foram discutidas. Os gêneros Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia e Achaeta foram identificados nas amostras.

Enchytraeid abundance in Araucaria Mixed Forest determined by cold and hot wet extraction / Abundância de enquitreídeos na Floresta Ombrófila Mista determinada por extração úmida quente e fria

Abstract Enchytraeids are small oligochaetes found worldwide in soils with sufficient moisture and organic matter, but scarcely studied in the Southern hemisphere. This is the third study on enchytraeid abundance in Brazil using wet extraction and the first carried out in Araucaria Mixed Forest (subtropical region). The sampling and extraction were based on the standard method ISO 23611-3/2007 using an adapted split soil corer and wet extraction with and without heat to assess the abundance of enchytraeids in a forest fragment at Embrapa Forestry in Colombo, Paraná State. The samplings were performed in 3 occasions between September 2011 and April 2012. The average numbers estimated by each method varied from appr. 2.000-12.000 (cold) and 5.000-12.000 ind./ m2 (hot), respectively, with a maximum of 44.000 ind./ m2 in one of the samples, the highest value reported so far in Brazil. The hot extraction was more advantageous, given the speed and preservation of the specimens in vivo, allowing taxonomic identification. Advantages and disadvantages of wet extractions compared to handsorting and formol methods are also discussed. Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia and Achaeta were the genera identified in the samples.

Resumo Os enquitreídeos são pequenos oligoquetas encontrados no mundo todo em solos com suficiente umidade e matéria orgânica, porém muito pouco estudados no hemisfério Sul. Este é o terceiro estudo sobre a abundância de enquitreídeos no Brasil utilizando o método de extração úmida e o primeiro realizado em Floresta Ombrófila Mista (região subtropical). A amostragem e extração foram baseadas no método padrão ISO 23611-3/2007, utilizando-se um trado desmontável adaptado e extração úmida com e sem aquecimento para acessar a abundância de enquitreídeo em um fragmento de floresta na Embrapa Florestas em Colombo, Paraná. As amostragens foram realizadas em três ocasiões entre setembro, 2011 e abril 2012. Os números médios estimados através de cada método variaram de 2.000-12.000 (frio) e 5.000-12.000 ind./ m2 (quente), respectivamente, e o máximo de 44.000 ind./ m2 em uma das amostras, o mais alto já relatado no Brasil. A extração quente foi a mais vantajosa, considerando a rapidez e preservação dos exemplares in vivo. As vantagens e desvantagens das extrações úmidas comparadas aos métodos de triagem manual e extração com formol foram discutidas. Os gêneros Guaranidrilus, Hemienchytraeus, Enchytraeus, Fridericia e Achaeta foram identificados nas amostras.

Recent Advances in Extraction Methods, Pharmacology,Toxicology and Pharmacokinetics Studies of Phill-yrin / 中国药师

Phillyrin is an important lignans compound from Forsythia. It is not only found in the raw medicinal material with high content, but also shows a variety of pharmacological activities. In the paper, the recent advances in extraction methods, pharmacology-toxicology, pharmacokinetics and the other studies of phillyrin were reviewed in order to provide reference and suggestions for the fur-ther research.

Comparison of three methods for the extraction of mycobacterial DNA / 中华皮肤科杂志

Objective To compare three methods for the extraction of mycobacterial DNA.Methods Two commercial DNA extraction kits and an ordinary freeze-thawing method were used to extract DNA from the pure suspensions of three species of Mycobacteria (M.tuberculosis,M.leprae and M.smegmatis) at different densities (1 × 10 to 1 × 105 cells/ml),simulated clinical specimens containing different concentrations of mycobacterial cells (1 × 10 to 1 × 104 cells/ml).The purity and concentration of the extracted DNA were evaluated.Then,PCR was performed to amplify the 16S rRNA region of Mycobacteria.The performance of the three methods was compared by the purity and concentration of extracted DNA as well as the results of PCR.Further more,76 clinical skin specimens suspected to be infected with Mycobacteria were used to further validate the performance of these methods.Results All the extracted DNA samples could be detected by PCR.The highest purity of DNA was obtained by the kit A,followed sequentially by the freeze-thawing method and the kit B.When pure suspensions were used,the detection limit was consistently 1 × 102 cells/ml for all the three methods.With simulated specimens,the detection rate was consistently 100％ for all the three methods at the concentration of 1 × 103 cells/ml,60％ (12/20),55％ (11/20) and 55％ (11/20) for the kit A,kit B and freeze-thawing method respectively at the concentration of 1 × 102 cells/ml.The analysis of clinical specimens showed that the kit B could be used to extract DNA from paraffin-embedded specimens,with the detection rate similar to that of kit A and freeze-thawing method.Conclusions The kit A could rapidly yield high-quality genomic DNA of Mycobacteria by repeated cleaning of columns,and may serve as the optimal method for scientific and clinical studies,and the kit B is suitable for extracting mycobacterial DNA from fresh tissue specimens besides paraffin-embedded specimens.

Evaluation of three different DNA extraction methods from blood samples collected in dried filter paper in Plasmodium subpatent infections from the Amazon region in Brazil / Avaliação de três métodos de extração de DNA obtidos de amostras de sangue coletadas em papel de filtro em infecções subpatentes de Plasmodium da região Amazônica no Brasil

Asymptomatic Plasmodium infection is a new challenge for public health in the American region. The polymerase chain reaction (PCR) is the best method for diagnosing subpatent parasitemias. In endemic areas, blood collection is hampered by geographical distances and deficient transport and storage conditions of the samples. Because DNA extraction from blood collected on filter paper is an efficient method for molecular studies in high parasitemic individuals, we investigated whether the technique could be an alternative for Plasmodium diagnosis among asymptomatic and pauciparasitemic subjects. In this report we compared three different methods (Chelex®-saponin, methanol and TRIS-EDTA) of DNA extraction from blood collected on filter paper from asymptomatic Plasmodium-infected individuals. Polymerase chain reaction assays for detection of Plasmodium species showed the best results when the Chelex®-saponin method was used. Even though the sensitivity of detection was approximately 66% and 31% for P. falciparum and P. vivax, respectively, this method did not show the effectiveness in DNA extraction required for molecular diagnosis of Plasmodium. The development of better methods for extracting DNA from blood collected on filter paper is important for the diagnosis of subpatent malarial infections in remote areas and would contribute to establishing the epidemiology of this form of infection.

Infecção assintomática por Plasmodium é um novo desafio para a saúde pública no Brasil. A reação em cadeia da polimerase (PCR) é o melhor método para detectar baixas parasitemias presentes em pacientes com infecção assintomática. Nas áreas endêmicas, a coleta de sangue total é dificultada pela distancia geográfica, transporte e adequada armazenagem das amostras. A coleta de sangue em papel de filtro pode ser uma alternativa nessas áreas de difícil acesso. Neste estudo foram comparados três diferentes métodos de extração de ADN a partir de papel de filtro usando como controle extração a partir de sangue total. O protocolo Chelex®-Saponina foi o que obteve o melhor resultado quando comparado com os outros três protocolos. No entanto a sensibilidade foi de 66,7% para o P. falciparum e 31,6% para o P. vivax. Conclui-se que em caso de infecção assintomática o papel de filtro não é ainda uma boa alternativa para coleta de amostras.

Bioactive compounds in Bidens pilosa L. populations: a key step in the standardization of phytopharmaceutical preparations

The total flavonoid content (TFC), total polyphenols content (TPC), and in vitro antioxidant activity (AA) of six Bidens pilosa L., Asteraceae, populations harvested from different localities were evaluated in this work. The plants were separated in roots, stems, and leaves/flowers, and the influence of extraction methods was investigated.Areversed-phase high-performance liquid chromatography method (HPLC) was developed and employed to obtain characteristic HPLC fingerprints of the bioactive compounds present in the extractive solutions, which were correlated with the TFC, TPC, and AA. Extractive solutions of leaves/flowers presented a higher AA when compared with those obtained from other parts of the plant (IC50 of 35.35±0.10 µg/mL). The stem extracts presented the lowest AA (IC50 117.2±1.96 µg/mL). A direct correlation of AA with TFC and TPC was evidenced. The highest AA was obtained by dynamic maceration and was statistically different from the AA presented by the extractive solutions obtained by other extraction methods. The results of this work evidenced differences that can be found at different stages of development of phytopharmaceutical preparations from B. pilosa and highlighted the importance of using the concentration of marker compounds as well as HPLC fingerprints as quality control parameters.