Effects of Pasturella Multocida B:2 and its immunogens (LPS and OMP) on reproductive hormones in Nili-Ravi Buffaloes

Abstract Livestock is a fundamental part of the agriculture industry in Pakistan and contributes more than 11.53% to GDP. Among livestock species, the buffaloes are regarded as the black gold of Pakistan. Being the highest milk producers globally, Nili-Ravi buffaloes are the most famous ones. Buffaloes are affected by many endemic diseases, and "Hemorrhagic septicemia" (HS) is one of them. This study was designed to ascertain the effects of experimental exposure ofP. multocida B:2 (oral) and its immunogens, i.e., LPS (oral and intravenous) and OMP (oral and subcutaneous) on reproductive hormonal profiles in Nili-Ravi buffaloes. Repeated serum samples were collected from the jugular vein of experimental animals for 21 days (0, 02, 04, 08, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 and 504 hours). Hormonal assays to determine the serum concentrations of Gonadotropin-releasing hormone (GnRH), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Estrogen (E2) and progesterone (P4) were performed using (MyBioSource) commercial Elisa kits. The hormonal profile of all treatment groups of the buffalo heifers exhibited significant (P 0.05) variations as compared to the control group (G-1). These results indicate suppression in Nili-Ravi buffaloes' reproductive hormonal profile on exposure to P. multocida B:2 and its immunogens. This influence warrants that exposure to H.S may be a possible reason for delayed puberty and poor reproduction performance in Nili-Ravi buffaloes.

Resumo A pecuária é uma parte fundamental da indústria agrícola no Paquistão e contribui com 11,53% do PIB nacional. Entre as espécies de gado, os búfalos são considerados o ouro negro do Paquistão. Sendo os maiores produtores de leite em todo o mundo, os búfalos Nili-Ravi são os mais famosos. Os búfalos são afetados por muitas doenças endêmicas, entre as quais a septicemia hemorrágica (SH). Este estudo busca verificar os efeitos da exposição experimental de P. multocida B:2 (oral) e seus imunógenos, ou seja, LPS (oral e intravenoso) e OMP (oral e subcutâneo), nos perfis hormonais reprodutivos em búfalos Nili-Ravi. Amostras de soro repetidas foram coletadas da veia jugular de animais experimentais por 21 dias (0, 2, 4, 8, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 e 504 horas). Os ensaios hormonais para determinar as concentrações séricas do hormônio liberador de gonadotrofina (GnRH), hormônio foliculoestimulante (FSH), hormônio luteinizante (LH), estrogênio (E2) e progesterona (P4) foram realizados usando kits comerciais Elisa (MyBioSource). O perfil hormonal de todos os grupos de tratamento das novilhas bubalinas apresentou variações significativas (P 0,05) em relação ao grupo controle (G-1). Esses resultados indicam supressão no perfil hormonal reprodutivo de búfalos Nili-Ravi na exposição a P. multocida B:2 e seus imunógenos. Essa influência garante que a exposição à SH possa ser uma possível razão para o atraso da puberdade e o baixo desempenho reprodutivo em búfalos Nili-Ravi.

Effects of Pasturella Multocida B: 2 and its immunogens (LPS and OMP) on reproductive hormones in Nili-Ravi Buffaloes / Efeitos de Pasteurella multocida B: 2 e seus imunógenos (LPS e OMP) em hormônios reprodutivos em búfalos Nili-Ravi

Abstract Livestock is a fundamental part of the agriculture industry in Pakistan and contributes more than 11.53% to GDP. Among livestock species, the buffaloes are regarded as the black gold of Pakistan. Being the highest milk producers globally, Nili-Ravi buffaloes are the most famous ones. Buffaloes are affected by many endemic diseases, and "Hemorrhagic septicemia" (HS) is one of them. This study was designed to ascertain the effects of experimental exposure ofP. multocida B:2 (oral) and its immunogens, i.e., LPS (oral and intravenous) and OMP (oral and subcutaneous) on reproductive hormonal profiles in Nili-Ravi buffaloes. Repeated serum samples were collected from the jugular vein of experimental animals for 21 days (0, 02, 04, 08, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 and 504 hours). Hormonal assays to determine the serum concentrations of Gonadotropin-releasing hormone (GnRH), Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Estrogen (E2) and progesterone (P4) were performed using (MyBioSource) commercial Elisa kits. The hormonal profile of all treatment groups of the buffalo heifers exhibited significant (P<0.05) variations as compared to the control group (G-1). These results indicate suppression in Nili-Ravi buffaloes' reproductive hormonal profile on exposure to P. multocida B:2 and its immunogens. This influence warrants that exposure to H.S may be a possible reason for delayed puberty and poor reproduction performance in Nili-Ravi buffaloes.

Resumo A pecuária é uma parte fundamental da indústria agrícola no Paquistão e contribui com 11,53% do PIB nacional. Entre as espécies de gado, os búfalos são considerados o ouro negro do Paquistão. Sendo os maiores produtores de leite em todo o mundo, os búfalos Nili-Ravi são os mais famosos. Os búfalos são afetados por muitas doenças endêmicas, entre as quais a "septicemia hemorrágica" (SH). Este estudo busca verificar os efeitos da exposição experimental de P. multocida B:2 (oral) e seus imunógenos, ou seja, LPS (oral e intravenoso) e OMP (oral e subcutâneo), nos perfis hormonais reprodutivos em búfalos Nili-Ravi. Amostras de soro repetidas foram coletadas da veia jugular de animais experimentais por 21 dias (0, 2, 4, 8, 12, 16, 20, 24, 36, 48, 72, 120, 168, 216, 264, 360, 456 e 504 horas). Os ensaios hormonais para determinar as concentrações séricas do hormônio liberador de gonadotrofina (GnRH), hormônio foliculoestimulante (FSH), hormônio luteinizante (LH), estrogênio (E2) e progesterona (P4) foram realizados usando kits comerciais Elisa (MyBioSource). O perfil hormonal de todos os grupos de tratamento das novilhas bubalinas apresentou variações significativas (P < 0,05) em relação ao grupo controle (G-1). Esses resultados indicam supressão no perfil hormonal reprodutivo de búfalos Nili-Ravi na exposição a P. multocida B:2 e seus imunógenos. Essa influência garante que a exposição à SH possa ser uma possível razão para o atraso da puberdade e o baixo desempenho reprodutivo em búfalos Nili-Ravi.

Quantitative estimation of enzymatic released specific oligosaccharides from Hericium erinaceus polysaccharides using CE-LIF / 药物分析学报

Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features.Therefore,quantitatively quality control of polysaccharides based on their chemical charac-teristics is important for their application in biomedical and functional food sciences.However,poly-saccharides are mixed macromolecular compounds that are difficult to isolate and lack standards,making them challenging to quantify directly.In this study,we proposed an improved saccharide mapping method based on the release of specific oligosaccharides for the assessment of Hericium eri-naceus polysaccharides from laboratory cultured and different regions of China.Briefly,a polysaccharide from H.erinaceus was digested by β-(1-3)-glucanase,and the released specific oligosaccharides were labeled with 8-aminopyrene-1,3,6-trisulfonic-acid(APTS)and separated by using micellar electrokinetic chromatography(MEKC)coupled with laser induced fluorescence(LIF),and quantitatively estimated.MEKC presented higher resolution compared to polysaccharide analysis using carbohydrate gel elec-trophoresis(PACE),and provided great peak capacity between oligosaccharides with polymerization degree of 2(DP2)and polymerization degree of 6(DP6)in a dextran ladder separation.The results of high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector(HPSEC-MALLS-RI)showed that 12 h was sufficient for complete digestion of polysaccharides from H.erinaceus.Laminaritriose(DP3)was used as an internal standard for quantifi-cation of all the oligosaccharides.The calibration curve for DP3 showed a good linear regression(R2＞0.9988).The limit of detection(LOD)and limit of quantification(LOQ)values were 0.05 μg/mL and 0.2 μg/mL,respectively.The recovery for DP3 was 87.32(±0.03)％in the three independent injections.To sum up,this proposed method is helpful for improving the quality control of polysaccharides from H.erinaceus as well as other materials.

Identification and activity evaluation of Astragalus Radix from different germplasm resources based on specific oligosaccharide fragments / 中草药·英文版

Objective: Based on trifluoroacetic acid (TFA) hydrolysis, polyacrylamide gel electrophoresis (PAGE) and high performance thin layer chromatography (HPTLC) analysis, the carbohydrate responsible for immunomodulatory activity are used as quality indicators for Astragalus Radix (AR). Methods: In this study, 24 batches of AR from different germplasm resources were selected as the research object, and AR polysaccharides were extracted. PAGE and HPTLC methods were used to analyze the partial acid hydrolyzate of AR polysaccharides and obtain a series of saccharide fingerprints. The data were analyzed by principal component analysis to obtain the difference between AR from different germplasm resources. Results: The results showed that trisaccharide and tetrasaccharide could be used as differential fragments to distinguish AR of different cultivation methods; Disaccharides and trisaccharides can be used as differential fragments to distinguish different species of AR. The immunological activity analysis of the specific oligosaccharide fragment of AR showed that the specific oligosaccharide fragment of AR could promote the secretion of TNF-α, IL-1β, IL-6, and NO in THP-1 cells in a concentration-dependent manner. Conclusion: Both PAGE and HPTLC methods can be used to evaluate AR from different germplasm resources. This study laid the foundation for the quality evaluation of AR medicinal herbs.

Fast saccharide mapping method for quality consistency evaluation of commercial xylooligosaccharides collected in China / 药物分析学报

Due to the extensive use of xylooligosaccharides (XOS) as functional food ingredients,many inferior goods and even adulterants are generally found in the market,which may pose a health hazard to certain populations.Chromatography method such as high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) is traditionally applied for the quality analysis of XOS.However,it is time consuming due to the prolonged separation and pre-or post-derivatization procedure.In this study,a fast saccharide mapping method based on matrix-assisted laser desorption/time-of-flight mass spectrometry (MALDI-TOF-MS) was developed for the quality consistency analysis of 22 batches of XOS collected from different manufacturers in China.The time needed for saccharides analysis using MALDI-MS was less than 30 min for one plate,at least 6 times faster than that by the traditional HPTLC chromatography method.In addition,MALDI-MS possessed higher resolution for XOS with DP4-DP7 based on the difference of m/z,which is hardly separated using HPTLC.The results showed that XOS were present only in samples XY01-XY11,samples XY12-XY14 only consisted of hex oligo-saccharides,and samples XY15-XY22 were free of oligosaccharides.These indicate that the quality consistency of XOS products in the China market was poor,which should be carefully investigated.

New Rabbit Erythrocyte Specific Mycelial Lectins from Fusarium sp. with Complex Saccharide Specificity

Eight Fusarium sp. namely, F. acutatum, F. globosum, F. graminearum, F. lactis, F. nivale, F. proliferatum, F.pseudoanthophilum and F. robustum were screened for the presence of lectins by hemagglutination activityusing human ABO, porcine, ovine, goat and rabbit erythrocytes. Mycelial extracts of all the fungal culturesexcept F. graminearum displayed unique lectin activity with only rabbit erythrocytes. Enzymatic treatment ofrabbit erythrocytes with neuraminidase has significantly enhanced the titre of all the lectin-positive extractsof fungal cultures. In contrast, most of the lectins showed a decline in lectin activity with protease treatedrabbit erythrocytes. Saccharide specificity studies have shown that majority of the lectins are inhibitory towardsO-acetyl sialic acids. None of the lectins from Fusarium sp. were inhibited by dextran, meso-inositol, andN-acetyl-D-glucosamine. Most of the fungal cultures displayed highest hemagglutination activity during the10th day of growth in broth cultures. The unique saccharide specificity of Fusarium sp. lectins can be used forelucidating their clinical role in glycobiology research.

Chemical Substantial Basis of Shengmai Injection(Ⅰ): Qualitative Analysis of Chemical Components / 中国药学杂志

OBJECTIVE： To conduct a comprehensive and systematic qualitative investigation on chemical components in Shengmai injection. METHODS： Gas chromatography-mass spectroscopy (GC-MS) with pre-column derivatization, rapid resolution liquid chromatography-ion trap-tandem mass spectroscopy (RRLC-IT-MSn) and ion exchange chromatography (IEC) with post-column derivatization were applied in identification of saccharides, saponins, lignins and amino acids in the sample. Furthermore, preliminary investigation on test of macromolecules was performed using size exclusion chromatography-evaporative light scattering detection (SEC-ELSD). RESULTS： Fructose, glucose, sucrose, maltose, 5-hydroxymethyl-2-furaldehyde, 19 saponins, 14 lignins and 16 amino acids were found in Shengmai injection. CONCLUSION： The proposed methods are simple and accurate. The main chemical substantial basis of Shengmai injection is clarified, laying a foundation for further assay of chemical components.

Correlation analysis between the chemical contents and bioactivity for the quality control of Alismatis Rhizoma

In order to clarify regions of production and to discriminate processing methods, quantitative and qualitative analyses for saccharides and terpenes in 35 batches of Alismatis Rhizoma were performed. Methodologies included HPLC-PDA, HPLC-VWD and UHPLC-MS , combined with principal component analysis (PCA) and partial least squares regression techniques (PLSR). The inhibitory effects of triterpenes and Alismatis Rhizoma extracts on lipase activity were evaluated . PLSR analysis revealed significant positive correlations ( = 0.5795) between the contents of triterpenes , , , and and the inhibitory effects of Alismatis Rhizoma. The present study establishes an effective method for simultaneous determination of multiple components, and identifies key bioactive triterpenes. These results can be used for systematic and novel analytical strategies for the quality control of Alismatis Rhizoma production.

Content Determination of Saccharide in Polysaccharides Containing Galacturonic Acid by Phenol-sulfuric Acid Method Combined with Calibration Factor Method / 中国药房

OBJECTIVE:To establish a method for the content determination of saccharide in polysaccharides containing galact-uronic acid based on phenol-sulfuric acid method combined with correction factor method. METHODS:The determination condi-tion of phenol-sulfuric acid was optimized. A series of standard curves were drawn with glacturonic aid-glucose mixed control with different mass ratio. The content of saccharide in Codonopsis pilosula polysaccharides CPP1b samples containing galacturonic acid was determined. According to regression equation of galacturonic acid-glucose ratio of 0-100%,the correction factor was calculated by using C. pilosula polysaccharides CPP1b as the reference polysaccharide,and the results of content determination of saccharide were corrected. The rationality of this method was verified by using mixed monosaccharide control with same composition as C. pi-losula polysaccharides CPP1b. RESULTS:The correction factor of C. pilosula polysaccharide CPP1b to glucose was 3.33;in vali-dation test,the content of saccharide in mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b was 103.47%. RSDs of precision and stability tests was <1%. The recoveries ranged 93.52%-107.35%(RSD=5.09%,n=6). CONCLUSIONS:The established method can accurately determine the content of saccharide in C. pilosula polysaccharides CPP1b containing galacturonic acid.

Effect of Codonopsis and Glycyrrhizae saccharide extracts on polyamine-dependent signaling pathway during cell migration in IEC-6 / 中国药理学通报

Aims To observe the effect of saccharide extracts of Yiqijianpi herb Codonopsis and Glycyrrhizae on polyamine-dependent activation of K+ channels sig-nal pathway during cell migration and to investigate their mechanism of promoting restoration in gastrointes-tinal mucosal injuries. Method The study was based on IEC-6 cell migration model. While in a normal polyamine level or polyamine was inhibited by DFMO, the effect of Codonopsis saccharide extracts and Glycyr-rhizae saccharide extracts on polyamine-dependent acti-vation of K+ channels signal pathway during cell mi-gration was observed. (1) K+ channel protein Kv1. 1 was determined by Western blot. (2)Membrane poten-tial was measured by Flow Cytometer. (3) Laser scan-ning confocal microscope was used for measuring [ Ca2+] cyt. ( 4 ) The expression of RhoA, which is Ca2+ downstream protein, was determined by Western blot. Results During cell migration, Codonopsis and Glycyrrhizae saccharide extracts could: ( 1 ) improve the expression of Kv1 . 1 protein and ameliorate the de-crease of kv1. 1 protein expression by DFMO;(2) in-crease membrane hyperpolarization and reverse mem-brane depolarization resulted by DFMO; ( 3 ) improve intracellular [ Ca2+] cyt, while Codonopsis could re-verse the decrease of [ Ca2+] cyt caused by DFMO;(4) improve the expression of RhoA protein, reversing its decline caused by DFMO. Conclusion Codonop-sis and Glycyrrhizae saccharide extracts can promote cell migration in IEC-6 cell, which is correlated with their effect on polyamine-dependent activation of K+channels signal pathway.

Effects of Hedysarum Polybotys Saccharide on Lipid Metabolism and Expression of Stearoyl-CoA Desaturase-1 Gene in NAFLD Rats / 中国中医药信息杂志

Objective To investigate the effects of hedysarum polybotys saccharide (HPS) on lipid metabolism and the expression of stearoyl-CoA desaturase-1(SCD-1) gene in rats with non-alcoholic fatty liver disease (NAFLD). To discuss the interfering effects of HPS on NAFLD. Methods The SD rats were randomly divided into the blank control group and the experiment group. Rats in the experiment group were fed with lipid rich food for 8 weeks to establish model and were randomly divided into model group, drug positive group and HPS group. After 8 weeks of drug intervention, the level of ALT, AST, TC, TG, HDL-c and LDL-c were measured with automatic chemistry analyzer, and expression of SCD-1 gene was measured by semi-quantitative polymerase chain reaction.Results Compared with blank control group, serum ALT, AST and TC, TG, LDL-c of model group were higher (P<0.05,P<0.01), the level of HDL-c of model group and the expression of SCD-1 gene were lower (P<0.01). Compared with model group, HPS was useful to decrease serum ALT, AST, LDL-c, TC and TG (P<0.05,P<0.01), and increase the level of HDL-c (P<0.01) and the expression of SCD-1 gene (P<0.01).ConclusionHPS had a positive effect on regulating lipid metabolic disturbance of NAFLD rats and promoting the expression of regulatory gene SCD-1.

Effects of lipopolysaccharide preconditioning on the activities of nuclear factor kappa-B and the expression of intercellular adhension molecule-1/lymphocyte function associated antigen-1 in rats graft with liver ischemia/reperfusion injury / 重庆医科大学学报

Objective: To study the effects and significances of lipopolysaccharide preconditioning on the activities of NF-?B and the expression of ICAM-1/LFA-1 in rats graft liver ischemia/reperfusion injury. Methods: Male Sprague-Dawley rats were divided into three groups: sham operation group(Sham group),orthotopic liver transplantation group (OLT group) and LPS preconditioning group (LPS group). Only dissecting hepatoduodenal ligament was perfomed in Sham group. Experiments of OLT were performed by two-cuff method in OLT group and LPS group. The activities of NF-?B and the expression of ICAM-1/LFA-1 in hepatic tissue ,the levels of ALT,AST in inferior caval vein blood were detected at 0,60,180 min after dissecting of hepatoduodenal ligament in Sham group and after portal vein reperfusion in OLT group and LPS group. Results: Compared with those in Sham group at the different time points respectively,the activities of NF-?B and the expression of ICAM-1/LFA-1 were higher in OLT group and LPS group(P0.05) at 0 min after reperfusion,they were evidently higher in OLT group than in LPS group (P

Study on the Influence of Cellulase on the Extraction Rate of Medlar Total Saccharide in Medlar by Sphere Symmetry Design / 中国中医药信息杂志

Objective To study the effect of cellulase on the extraction rate of medlar total saccharide, considering three factors of temperature, time and quantity of cellulose. The best extracting condition was determined according to the require of sphere symmetry design. Methods Add cellulase liquid into the medlar slurry and determine the content of medlar total saccharide by DNS method. Result The extraction rate of medlar total saccharide was increased 1.66% by cellulase at temperature of 60 ℃, time of 300 s and cellulase volume of 5 mL. Conclusion Cellulase can promote the extraction rate of medlar total saccharide.

Effect of Hedysarum Polybotys Saccharide on Insulin Resisance of Experimental NIDDM Rats / 中国中医药信息杂志

Objective To investigate the effect of Hedysarum Polybotys saccharide(HPS)on insulin resistance rats.Methods The model of type 2 diabetic-insulin resistance rat was successfully made by STZ and high caloric diet.Then the rats were intervene by Metformin(MT)and HPS.Results Compared with the model group,the level of blood glucose,INS and CP were lowered,ISI were improved and FFA were restrained by HPS.Conc(?)sion HPS can reduce the insulin resistance in multiple ways.

Componentes glicosilados de la envoltura de Mycobacterium tuberculosis que intervienen en la patogénesis de la tuberculosis / Structural components of the envelope of Mycobacterium tuberculosis that intervene in the pathogenesis of tuberculosis

Los componentes glicosilados de la envoltura de Mycobacterium tuberculosis tienen un papel importante en la inmunopatogénesis de la tuberculosis. Permiten la adhesión, penetración y persistencia de la micobacteria en el macrófago; de igual manera, participan en los mecanismos de activación de estas células y la producción de citocinas relevantes durante la respuesta inmune. En esta revisión, examinamos las características de las principales estructuras sacarídicas de la superficie de la micobacteria y su relación con la modulación de la respuesta inmune.

The glycosylated compounds of Mycobacterium tuberculosis envelope play an important role in the immunopathogenesis of tuberculosis. These molecules are involved in the binding to the host cell surface followed by their internalization and persistence in macrophages; likewise they take part in the macrophage's activation and the production of cytokines that are relevant during the immune response against mycobacteria. In this review we examine the molecular characteristics of the main mycobacterial cell surface saccharide structures and their relation with the modulation of the immune response to tuberculosis.

The cytokine-inducing activities of surface components of the periodontopathogenic bacterium Porphyromonas gingivalis / 대한치주과학회지

This study was carried out to examine the potency of the three surface components from Porphyromonas gingivalis to stimulate the murine macrophage cell line RAW264.7 to synthesize the pro-inflammatory cytokine tumor necrosis factor alpha(TNF-alpha) and nitric oxide (NO). Lipopolysaccharide(LPS), lipid A-associated proteins(LAP) and saline-extractable surface -associated material(SAM) were isolated from P. gingivalis 381. TNF-alpha release into culture supernatants was determined by two-site ELISA. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. Western blot analysis of iNOS and analysis of reverse transcription(RT)-PCR products were carried out. The surface components extracted from this bacterium were almost equally potent in stimulating release of TNF-alpha and NO by RAW264.7 cells. TNF-alpha that was being measured immunologically was due to activation of TNF-alpha gene transcription. The present study clearly shows that P. gingivalis surface components fully induced iNOS expression in RAW264.7 cells in the absence of other stimuli. The ability of P. gingivalis surface components to promote the production of TNF-alpha and NO may be important in the pathogenesis of inflammatory periodontal disease.

Investigation progress on receptors of saccharides / 中国药理学通报

More and more interest has been put on the investigations of saccharides recent years. Saccharides have a vast number of bioactivities and the immunomodulating effect of saccharides is one of the important mechanisms via which saccharides exert their bioactivities. Saccharides exert their bioactivities mainly by interacting with all kinds of saccharide receptors on cells, followed by a series of signal transduction process. In the present paper, numerous saccharide receptors and the characters of the mutual interaction between saccharides and their receptors are summarized.

Effect of Tripterygtam wilfordii Glycosides and poly saccharide sulphate on the intimal proliferation of injured rat aorta / 中国介入心脏病学杂志

31 rat were divided randomly into control group (n=10),tripterygium wilfordii glycosides group (n = 10 9. 4 mg/kg per day),and poly saccharide sulphate group (n=11. 30 mg/per day). We made balloon endothelium denudation in thoracic aortae of the rats. The treatments with drugs began 6 days before balloon injury and continously until the animal were killed 14 day after balloon injury. The determinations for area of neointima, neointima /media, coverage of neointima and 3H-TdR incoperation of thoracic aortae suggested that tripterygium wilfordii glycosides can inhibit intimal proliferation of injuried aorta, whill poly saccharide sulphate shows no effect.

Determination of monosaccaride and disaccaride in condensed Siwu Decoction by HPLC-ELSD / 中草药

Object To establish a method by high performance liquid chromatography/evaporative light scattering detector (HPLC-ELSD) for the determination of contents of D-fructose, D-glucose, and sucrose in Siwu Decoction (SWD). Methods Condensed SWD was analyzed by HPLC-ELSD after dilution, precipitation by ethanol. Results Condensed SWD, 1 mL, contained (33.4?1.5) mg of D-fructose, (24.2?0.9) mg of D-glucose and (112.7?6.1) mg of sucrose. The standard curves were linear within the range of 0.15—3.75 mg/mL for D-fructose and 0.15—5 mg/mL for D-glucose and sucrose. The recovery rates were 128.5% for D-fructose, 114.7% for D-glucose, and 124.7% for sucrose. The relative standard deviations (RSD) within-days were 3.0% for D-fructose, 3.2% for D-glucose, and 4.4% for sucrose. Conclusion SWD contains abundant monosaccharide and disaccharide. HPLC-ELSD can be used to analyse the monosaccharide and disaccharide in SWD.

Binding of 4-methyl umbelliferyl-α-D-glucopyranoside to Vicia faba lectin: Fluorescence-quenching studies.

On binding to Vicia faba lectin, the fluorescence of 4-methylumbelliferyl-α-Dglucopyranoside was quantitatively quenched showing that the interaction of 4- methylumbelliferyl-α-D-glucopyranoside took place in a binding environment. The binding of the fluorescent sugar was saccharide specific as evidenced by the reversal of 4-methylumbelliferyl- α-D-glucopyranoside fluorescence quenching by D-fructose. The association constant, Ka, values for the 4-methylumbelliferyl-α-D-glucopyranoside was determined by competition study employing reversal of fluorescence quenching of 4-methylumbelliferyl-α- D-glucopyranoside by D-fructose. The Ka value obtained for D-fructose was 1·07 ± 0·03 × 104 M–1 and for 4-methylumbelliferyl-α-D-glucopyranoside was 1·60 ± 0·05 × 104 M–1 at 15°C. The Ka values of 2·51 ± 0·06 × 104 M–1, l·26 ± 0·02 × 104 Μ–1 and 0·56 ± 0·01 × 104 M–1, respectively at 10°, 20° and 30°C were obtained from the Chipman equation. The relative fluorescence quenching, Δ Fa, at infinite concentration of the free saccharide sites of Vicia faba lectin [P’] was 93·5% at 30°C and the binding constant for 4-methylumbelliferyl-α-Dglucopyranoside lectin interaction as derived by Yank and Hanaguchi equation was 0·63 ± 0·01 × 104 M–1.