Leche materna deslactosada con la enzima beta galactosidasa para lactantes intolerantes / Leite materno sem lactose com a enzima beta-galactosidase para lactentes intolerantes / Lactose-free breast milk with the enzyme beta galactosidase for intolerant infants

Introducción: los bebés deben beneficiarse de la leche materna, incluso cuando presentan intolerancia a la lactosa. Por esto, se debe recurrir a la obtención de leche materna deslactosada. Objetivo: analizar el efecto de la enzima beta galactosidasa en la hidrólisis de la lactosa de leche materna madura para bebés clínicamente diagnosticados con intolerancia a la lactosa. Materiales y método: estudio exploratorio, descriptivo y explicativo. El contenido de lactosa se cuantificó desde el inicio hasta el final del tratamiento, controlando temperatura, tiempos y cantidad de enzima ß-galactosidasa adicionada en la leche materna. Se recolectaron 1000 ml de leche materna, obtenidos del Banco de Leche del Hospital General de Medellín (Antioquia, Colombia). Resultados: las muestras donadas se encontraban pasteurizadas y posteriormente fueron sometidas a la acción de la enzima lactasa. Se cuantificó el contenido de lactosa sin la enzima, reportando en promedio 6,34 mg/100 ml ± 0,23. El mayor aporte de lactosa obtenido posterior a la exposición a la enzima (30 minutos) fue de 6,07 mg/ml ± 0,35 (correspondiente a 95 % del contenido inicial), finalizando con un aporte de 0,35 % a una concentración de 0,4 % tras 24 horas, porcentaje que representa 95 % de la hidrólisis total en la leche materna. Conclusiones: en todas las muestras analizadas de diferentes madres se pudo obtener leche materna con bajas concentraciones de lactosa tras 24 horas de haber sido sometidas a la acción de ß-galactosidasa. Lo anterior se establece como una alternativa para los bebés intolerantes a la lactosa, que permitiría no privarlos de todos los beneficios que ofrece este alimento.

Introduction: Babies should benefit from breast milk, even when they are lactose intolerant. For this reason, parents should resort to obtaining lactose-free breast milk. Objective: To examine the effect of the enzyme ß-galactosidase on the hydrolysis of lactose in mature breast milk for babies clinically diagnosed with lactose intolerance. Materials and method: Exploratory, descriptive, and explanatory study. The lactose content was quantified from the beginning to the end of the treatment, controlling variables such as temperature, times, and the amount of ß-galactosidase enzyme added in breast milk. A total of 1000 ml of breast milk were obtained from the milk bank at Hospital General de Medellín (Antioquia, Colombia). Results: Donated samples were first pasteurized and subsequently subjected to the action of the enzyme lactase. The lactose content without the enzyme was quantified, reporting an average of 6.34 mg/100 mL±0.23. The highest contribution of lactose obtained after exposure to the enzyme was 6.07 mg/mL±0.35 (corresponding to 95% of the initial content), at 30 minutes, ending with a contribution of 0.35% at a concentration of 0.4% in 24 hours, percentage that represents 95% of total hydrolysis in breast milk. Conclusions: In all the examined samples from different mothers, it was possible to obtain breast milk with low concentrations of lactose 24 hours after these were exposed to the action of ß-galactosidase. This becomes an alternative for feeding lactose intolerant babies and not deprive them from all the benefits offered by breast milk.

Introdução: os bebês se devem beneficiar do leite materno, mesmo quando tenham intolerância à lactose, razão pela qual se deve recorrer à obtenção de leite materno sem lactose. Objetivo: analisar o efeito da enzima beta-galactosidase na hidrólise da lactose no leite materno maduro para bebês diagnosticados clinicamente com intolerância à lactose. Materiais e método: estudo exploratório, descritivo, explicativo. O teor de lactose foi quantificado do início ao fim do tratamento; temperatura, tempos e quantidade de enzima beta-galactosidase adicionada no leite materno foram controlados; foram coletados 1000ml de leite materno, obtidos no Banco de Leite do Hospital General de Medellín (Antioquia, Colômbia). VResultados: as amostras doadas foram pasteurizadas e posteriormente submetidas à ação da enzima lactase. O teor de lactose sem a enzima foi quantificado, relatando uma média de 6,34mg/100ml±0,23. A maior contribuição de lactose obtida após a exposição à enzima foi de 6,07mg/ml±0,35 (correspondendo a 95% do conteúdo inicial) em 30 minutos, finalizando com uma contribuição de 0,35% na concentração de 0,4% em 24 horas, percentual que representa 95% da hidrólise total no leite materno. Conclusões: em todas as amostras analisadas de diferentes mães, foi possível obter leite materno com baixas concentrações de lactose 24 horas após ser submetido à ação da beta galactosidase, como alternativa para bebês intolerantes à lactose e não os privar de todos os outros benefícios oferecidos por esse alimento ideal.

Analysis of Transport Mechanism of Flavanomarein in MDCK Monolayer Cell Model / 中国实验方剂学杂志

Objective:To explore the absorption and transport properties of flavanomarein in the Madin-Darby canine kidney(MDCK) monolayer cell model. Method:Methyl thiazolyl tetrazolium(MTT) assay was used to investigate the toxicity of flavanomarein in MDCK cells. The resistance value of MDCK monolayer cell model was detected by Millicell-ERS-2 cell resistometer. The effects of mass concentration of flavanomarein,administration time,sodium-glucose cotransporter(SGLTs) inhibitor and glucose transporter 2(GLUT2) inhibitor on the transmembrane transport of flavanomarein were investigated. The concentration of flavanomarein was determined by UPLC-MS/MS, and the apparent permeability coefficient(Papp) and the efflux ratio(ER) were calculated. Result:When the concentration of flavanomarein was 5.625-120 mg·L-1, there was no significant toxic effect on MDCK cells. The transport of flavanomarein in MDCK monolayer cell model was time-dependent and concentration-dependent. The Papp values of flavanomarein were basically between 1×10-6 cm·s-1 to 10×10-6 cm·s-1. Compared with the blank group, the phlorizin group significantly reduced the transport of flavanomarein on the MDCK monolayer cell model at 60 min and 90 min. Conclusion:Flavanomarein is a moderately absorbed drug in the intestine, its transmembrane transport mechanism is dominated by passive transport along with active transport. SGLTs may be involved in mediating the transport of flavanomarein on the MDCK monolayer cell model.

Comparison of Improving Effects for Diabetic Erectile Dysfunction according to the Anti-Glycemic Agents: Phlorizin and Insulin

PURPOSE: To compare the improving effects of diabetic erectile dysfunction with two anti-glycemic agents; phlorizin and insulin. MATERIALS AND METHODS: Sixty Sprague-Dawley rats were divided into four groups (n=15 in each group): normal control (C), untreated diabetic rats (D), and diabetic rats treated by phlorizin (P) or insulin (I). Ten weeks after the diabetic induction using an injection of streptozotocin (55 mg/kg), four weeks of diabetic control was conducted. Erectile response, Western blot, and immunohistochemistry were assessed. RESULTS: During the experiment, the C-group showed continuous weight gain, while the other groups suffered from weight loss. After start of diabetic control, the body weight of I-group was increased; whereas, there was no meaningful change in the P-group. Meanwhile, comparable blood glucose levels were achieved in the P- and I-groups. The erectile response was markedly decreased in the D-group, whereas the P- and I-groups were similar as good as the C-group. In addition, D-group showed the significant decrease in the cavernosal smooth muscle content and increased apoptosis. Platelet endothelial cell adhesion molecule-1 protein expression, phosphorylation of endothelial nitric oxide synthase and myosin phosphatase target subunit 1 were significantly distorted in the D-group, while the P- and I-groups were comparable with the C-group. CONCLUSIONS: Phlorizin treatment resulted in the improvement of erectile function as same as insulin despite the lack of anabolic weight gains. These results suggest that control of blood glucose level rather than a type of anti-glycemic agents is more important for the prevention and treatment of diabetic erectile dysfunction

Metabolomics analysis and quantitative determination of five components in Malus leaves / 中草药

Objective The aim of this experiment is to compare the chemical composition of Malus leaves from different origins, which can be as a fundamental research for the comprehensive utilization and resource development. Methods A UPLC-DAD method has been developed for the determination of five compounds (phlorizin, trilobatin, phloretin, rutin, and quercitrin) in Malus leaves, and UPLC-QTOF-MS based metabonomics was applied to the comparison among different groups from different origins and species. Results There were significant difference in the content of five compounds, of which phlorizin was widely contained and its content was relatively high in the range of 1.01% -17.15%. Phloretin, rutin, and quercutrin were also widespread, while the trilobatin was only in Ser. Baccatae (Rehd.) Rehd and Sect. Sorbomalus Zabel, which was not detected in the cultivating-varieties. The results of metobonomics showed that the chemical composition in leaves of Malus between Ser. Pumilae (Rehd). Rehd and Ser. baccatae (Rehd). Rehd were clearly distinguished, and 26 discrimination markers as the basis for distinction were identified; The chemical composition of Malus leaves from different sources of origins were different while the wild species were similar to the cultivating- varieties. Conclusion The Malus leaves generally contain phlorizin and other polyphenols, which are the potential drugs and health products resources; There is certain difference in chemical composition and content of Malus leaves, which provide the scientific basis for the next utilization; Malus leaves are so rich and renewable that are worthy of further research and development.

Research progress on traditional Chinese medicine against diabetes based on intestinal flora new targets / 药物评价研究

Diabetes has become the third largest killer to human life and health,and its development is closely related to the intestinal flora imbalance caused by series of consequences.So adjusting the structure of intestinal flora becomes a new idea for the treatment of diabetes.Traditional Chinese medicine (TCM) has good curative effect in prevention and treatment of diabetes,and improvement on the structure of intestinal flora may be one of its important mechanisms.The article reviewed the research progress on prevention and treatment of diabetes by TCM single components (berberine,phloridzin,rheinic acid,etc),TCM single prescription (essential oil from Cinnamomum cassia,flower stalk of Coptis chinensis,total coumarin of Feucedani Radix) and TCM priscriptions (Gegen Qinlian Decoction,Shengjiang Powder,Buzhong Yiqi Decoction,Huanglian Jiedu Decoction,Wenyang Yiqi Huoxue Compound,etc.) based on new targets in intestinal flora,so as to provide reference for the development of TCM for prevention and treatment of diabetes.

Research progress on traditional Chinese medicine against diabetes based on intestinal flora new targets / 药物评价研究

Diabetes has become the third largest killer to human life and health,and its development is closely related to the intestinal flora imbalance caused by series of consequences.So adjusting the structure of intestinal flora becomes a new idea for the treatment of diabetes.Traditional Chinese medicine (TCM) has good curative effect in prevention and treatment of diabetes,and improvement on the structure of intestinal flora may be one of its important mechanisms.The article reviewed the research progress on prevention and treatment of diabetes by TCM single components (berberine,phloridzin,rheinic acid,etc),TCM single prescription (essential oil from Cinnamomum cassia,flower stalk of Coptis chinensis,total coumarin of Feucedani Radix) and TCM priscriptions (Gegen Qinlian Decoction,Shengjiang Powder,Buzhong Yiqi Decoction,Huanglian Jiedu Decoction,Wenyang Yiqi Huoxue Compound,etc.) based on new targets in intestinal flora,so as to provide reference for the development of TCM for prevention and treatment of diabetes.

Therapeutic effects of phlorizin on the early stage of diabetic retinopathy in diabetic db/db mice / 中华老年医学杂志

Objective To investigate therapeutic effects of phlorizin on the early stage of diabetic retinopathy(DR)in diabetic db/db mice.Methods The 24 db/db male mice were divided into 3 groups of phlorizin-treated diabetic group(20mg/kg of phlorizin by intragastric gavage)(DMT,n =8),diabetic group(normal saline solution by intragastric gavage)(DM,n=8),with C57BLKS/J db/ m mice as the control group(CC,n=8)for ten weeks.After ten weeks,all mice were sacrificed and the retina was isolated.The specimen was embedded in paraffin wax and sliced.The sections were dyed with Sudan red and observed for retinal morphological change.Meanwhile,the expressions of GSK-3β and phospho-GSK-3β in retinas were determined by western blot.Results During the study,the CC group showed good condition.The DM group featured with polydipsia,polyphagia and a rapid increase of body weight.Phlorizin-treated diabetic mice displayed much better than the DM group.Compared with CC group,DM group showed cells proliferation in capillary endothelia and obvious thickening in basement membrane.After administration of phlorizin,the vascular lesion of central regions was remarkably improved.The expression level of phospho-GSK-3β protein was lower in DM group than in control group(76.2±15.8 vs.255.4 ± 10.7,t =16.30,P＜0.01)and was higher in Phlorizin-treated DM group(188.5±18.4)than in DM group(76.2± 15.8,t=8.05,P＜0.01).Conclusions Phlorizin decreases phospho-GSK-3β3expression in retina in db/db diabetic mice and suppresses the GSK-3βactivity,which leads to the protection of DR.

Seasonal Variation, Microscopic and Chromatographic Analysis of Leaves in Malus hupehensis: A Protocol for Its Quality Control / 中草药·英文版

Objective: To establish a quality control protocol based on microscopic, TLC, and HPLC methods, and to verify the optimal harvesting time for the leaves of Malus hupehensis (LMH). Methods: The LMH were pulverized into powder for microscopic identification or TLC and HPLC analysis after ultrasonic extraction with methanol. Seasonal variations of the phlorizin content and average leaf weight were determined by HPLC analysis and weighing up the leaves collected from May to October. Results: Microscopic and macromorphologic characteristics have been described for the leaf identification. A qualitative TLC assay and a quantitative HPLC method have been established for the quality control of LMH. Phlorizin was selected as a reference marker, which resolved at Rf 0.53 in TLC assay and at 14.0 min in HPLC assay. The content of phlorizin decreased gradually from 17.0% in leaves collected in May to 7.5% in October. The average leaf weight reached the level of 0.6 g in August and maintained until its falling. Conclusion: These methods are simple, selective, accurate, and reliable for the quality control of LMH. The period from late August to early September is suggested as the optimal harvesting time of the LMH. © 2013 Tianjin Press of Chinese Herbal Medicines.

SGLT-2 inhibitors: The glucosuric antidiabetics.

Despite availability of a number of oral antidiabetics, a sizeable population of diabetics remains uncontrolled. Thus there is growing need of new group of drugs for diabetic control. Understanding renal conservation of glucose by efficient reabsorption through sodium glucose cotransporter-2 (SGLT-2) has paved way for development of an entirely new group of drugs, the SGLT-2 inhibitors. These glucosuric antidiabetic agents have shown promise in early clinical studies. Canagliflozin is recently approved for use in diabetes alone or along with other antidiabetics. Other highly selective inhibitors undergoing various stages of clinical developments are dapagliflozin, sergliflozin, remogliflozin, ipragliflozin, empagliflozin, luseogliflozin, tofogliflozin and desoxyrhaponticin. KGA-2727 (pyrazole-O-glucoside) is the first selective SGLT-1 inhibitor undergoing intense preclinical testing. There are safety issues associated with this group like urogenital infections (fungal), weight loss, initial osmotic diuresis and increased incidence of cardiovascular events. The long term safety remains to be established. Despite these limitations, SGLT-2 inhibition offers a unique target for achieving adequate control of diabetes in adults.

Effects of phlorizin on aorta of diabetes db/db mice / 中华老年医学杂志

Objective To observe the effects of phlorizin on aorta of diabetes db/db mice and to explore its mechanism.Methods Sixteen diabetes db/db mice were randomized into two groups:untreated diabetic group (DM group) and diabetic group treated by phlorizin(DMT group) with eight db/m mice as normal control group(CC group).Phlorizin (20 mg/kg) was given in normal saline solution intragastrically to the mice of the DMT group for 10 weeks.Mice of the other two groups were given the same amount of saline solution intragastrically for 10 weeks.Animals were weighed weekly.At 10th weekend,all mice were fasted overnight and then sacrificed.Fasting blood was collected,and aortas were dissected.The blood samples were analyzed for fasting blood glucose (FBG),triglycerides (TG),total cholesterol (TC),Serum advanced glycation end products(AGEs)and malondialdehyde (MDA).Aortic tissue were examined microscopically.Results At 10th weekend,the weight (57.53±3.40)g and serum concentration of FBG(31.21±2.16) mmol/L,TG (0.39±0.12) mmol/L,TC(3.15±0.30)mmol/L,AGEs (0.28±0.04) AU/mg and MDA (15.18± 1.60)mol/L in DM group were increased than those in CC group (P＜0.01),and the weight(54.24± 1.28)g,FBG(29.17±1.41) mmol/L,TG(0.26±0.06) mmol/L,TC(2.71±0.26) mmol/L,AGEs (0.24±0.03) AU/mg and MDA(13.46±1.28)mol/L were lowered significantly in DMT group than those in DM group (P＜0.05 or P＜0.01).The severity of aorta damage in DMT group was less than that in DM group.Conclusions Phlorizin can protect the db/db mice from diabetic macrovascular complications,which may be attributed to its decreasing of blood glucose,TG,TC,and AGEs levels,and its antioxidant potential.

Correlation between lactose absorption and the C/T-13910 and G/A-22018 mutations of the lactase-phlorizin hydrolase (LCT) gene in adult-type hypolactasia

The C/T-13910 mutation is the major factor responsible for the persistence of the lactase-phlorizin hydrolase (LCT) gene expression. Mutation G/A-22018 appears to be only in co-segregation with C/T-13910. The objective of the present study was to assess the presence of these two mutations in Brazilian individuals with and without lactose malabsorption diagnosed by the hydrogen breath test (HBT). Ten milk-tolerant and 10 milk-intolerant individuals underwent the HBT after oral ingestion of 50 g lactose (equivalent to 1 L of milk). Analyses for C/T-13910 and G/A-22018 mutations were performed using a PCR-based method. Primers were designed for this study based on the GenBank sequence. The CT/GA, CT/AA, and TT/AA genotypes (lactase persistence) were found in 10 individuals with negative HBT. The CC/GG genotype (lactase non-persistence) was found in 10 individuals, 9 of them with positive HBT results. There was a significant agreement between the presence of mutations in the LCT gene promoter and HBT results (kappa = -0.9, P < 0.001). The CT/AA genotype has not been described previously and seems to be related to lactase persistence. The present study showed a significant agreement between the occurrence of mutations G/A-22018 and C/T-13910 and lactose absorption in Brazilian subjects, suggesting that the molecular test used here could be proposed for the laboratory diagnosis of adult-type primary hypolactasia.

Study on Quality Standard of Lithocarpus polystachys / 中国药房

OBJECTIVE: To establish the quality standard of Lithocarpus polystachys.METHODS: TLC was used for the qualitative identification of L.polystachys.The content of total flavonoids of L.polystachys was determined by UV spectrophotometry and the content of phloridzin was determined by HPLC.RESULTS: The TLC spots of polyamide were clear and well separated.The maximum absorption wavelength of phlorizin was 284 nm.The content of total flavonoids of 6 batches of L.polystachys ranged from 103.12 mg?g-1 to 183.54 mg?g-1.The linear ranges of phlorizin were 0.099 8～1.197 6 ?g (r=0.999 7) with an average recovery of 97.23%(RSD=1.57%,n=6).CONCLUSION: Established quality standard is applicable for the quality control of L.polystachys.

Effect of cisplatin on sodium-dependent hexose transport in LLC-PK-1 renal epithelial cells

Cis-dichlorodiammine platinum II (Cisplatin), an effective chemotherapeutic agent, induces acute renal failure by unknown mechanisms. To investigate direct toxic effects of cisplatin on the renal proximal tubular transport system, LLC-PK-1 cell line was selected as a cell model and the sugar transport activity was evaluated during a course of cisplatin treatment. Cells grown to confluence were treated with cisplatin for 60 min, washed, and then incubated for up to 5 days. At appropriate intervals, cells were tested for sugar transport activity using alpha-methyl-D-(14C)glucopyranoside (AMG) as a model substrate. In cells treated with 100 micrometer cisplatin, the AMG uptake was progressively impaired after 3 days. The viability of cells was not substantially changed with cisplatin of less than 100 micrometer, but it decreased markedly with 150 and 200 micrometer. In cisplatin-treated cells, the Na+/-dependent AMG uptake was drastically inhibited with no change in the Na+/-independent uptake. Kinetic analysis indicated that Vmax was suppressed, but Km was not altered. The Na+/-dependent phlorizin binding was also decreased in cisplatin-treated cells. However, the AMG efflux from preloaded cells was not apparently retarded by cisplatin treatment. These data indicate that the cisplatin treatment impairs Na+/-hexose cotransporters in LLC-PK-1 cells and suggest strongly that defects in transporter function at the luminal plasma membrane of the proximal tubular cells constitute an important pathogenic mechanism of cisplatin nephrotoxicity.

Lactase mRNA Expression in Small Intestines of Korean Fetuses and Adults / 대한내과학회지

OBJECTIVE: The specific activity of lactase-phlorizin hydrolase (LPH) is very high at birth and sharply declines after weaning, producing lactose intolerance. The prevalence of lactose intolerance is up to 85% in Korean adults. Molecular basis of the regulatory mechanisms responsible for the decline of LPH specific activity is still unknown. In order to elucidate the molecular mechanisms regulating the LPH expression during development, LPH specific activity and mI4NA level of Korean fetal and adult intestines were compared. METHODS: 20 fetal small intestines (16-27 weeks) were obtained during therapeutic abortion and were divided into 3 equal length. 20 adult jejunal tissues were obtained from patients without small intestinal disease during laparotomy. Mucosal homogenates were prepared for dissacharidases specific activities measurement and total RNA was extracted for northern and slot hvbridization. LPH mRNA level was measured by laser densitometer. RESULTS: LPH specific activities of proximal, middle and distal portion of fetal intestines (n=20) were 36.2 +/- 22.5, 38.6 +/- 23.2 and 23.2 +/- 19.9 mu/mg protein, respectively. LPH specific activity of adult jejunum (n=8) was 5.9 +/- 1.8 mu/mg protein and significantly (p<0.05) lower than those of fetal intestines. However, there was no significant difference in sucrase and trehalase specific activities between fetal intestines and adult jejunum. Although LPH specific activity of adult jejunum was lower than those of fetal intestines, LPH mBNA level of adult jejunum was as high as those of fetal intestines. CONCLUSION: These results show that LPH specific activity and mRNA level do not parallel, indicating the posttranscriptional control of fetal development of LPH expression.