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Objective To evaluate the effect and mechanism of terminal fucosylation inhibitor 2-deoxy-D-galactose (2-D-gal) on ciclosporin (CsA)-induced renal epithelial-mesenchymal transition (EMT). Methods Fifteen male C57BL/6 mice aged 8-10 weeks were randomly and evenly divided into the control group (Ctrl group), CsA group and CsA+2-D-gal group (n=5). The expression levels of fucosyltransferase 1 (FUT1), EMT-associated proteins including E-cadherin, Vimentin, α-smooth muscle actin (α-SMA) in the kidney tissues of the Ctrl and CsA groups were detected by Western blot. The expression levels of terminal fucose in the kidney tissues of Ctrl and CsA groups were determined by immunofluorescence. The renal fibrosis of mice in each group was evaluated by Masson staining. The blood urea nitrogen and serum creatinine levels of mice in each group were detected. The in vitro EMT model of renal tubular epithelial cell HK2 was induced by CsA. HK2 cells were stimulated with 0, 2.5, 5.0 and 10.0 μmol/L CsA for 24 h, respectively. In addition, HK2 cells were divided into the Ctrl, 2-D-gal, CsA and CsA+2-D-gal groups. The morphology of HK2 cells after stimulation with different concentrations of CsA and in each group was observed. The expression levels of FUT1, E-cadherin, Vimentin and α-SMA in HK2 cells after stimulation with different concentrations of CsA and in each group were detected by Western blot. The expression level of terminal fucose in HK2 cells of the Ctrl and CsA groups was measured by immunofluorescence. Results Compared with the Ctrl group, the relative expression of E-cadherin protein was down-regulated, those of FUT1, Vimentin and α-SMA proteins were up-regulated (all P < 0.05), and that of terminal fucose in the mouse kidney tissues was up-regulated in the CsA group. Compared with the Ctrl group, the blood urea nitrogen and serum creatinine levels in the CsA and CsA+2-D-gal groups were up-regulated (all P < 0.05). Compared with the CsA group, the blood urea nitrogen and serum creatinine levels in the CsA+2-D-gal group were down-regulated (both P < 0.05). Compared with the Ctrl group, the collagen fiber deposition was increased and the relative expression of α-SMA protein was up-regulated in the mouse kidney tissues of CsA and CsA+2-D-gal groups (all P < 0.05). Compared with the CsA group, the collagen fiber deposition was decreased and the relative expression of α-SMA protein in the mouse kidney tissues was down-regulated in the CsA+2-D-gal group (both P < 0.05). With the increase of CsA concentration, the morphology of HK2 cells gradually became longer and thinner from original normal cobblestone shape, the relative expression levels of FUT1, Vimentin and α-SMA protein in HK2 cells were up-regulated, and that of E-cadherin protein was down-regulated in a concentration-dependent manner. Compared with the Ctrl group, the expression level of terminal fucose of HK2 cells was up-regulated in the CsA group. After CsA treatment combined with 2-D-gal intervention, the morphology of HK2 cells in the CsA+2-D-gal group was restored to resemble that of normal HK2 cells. Compared with the CsA group, the relative expression of E-cadherin protein in HK2 cells was up-regulated, whereas those of Vimentin and α-SMA proteins were down-regulated in the CsA+2-D-gal group (all P < 0.05). Conclusions CsA may induce EMT both in vivo and in vitro, and the terminal fucosylation is increased. 2-D-gal may inhibit CsA-induced EMT by suppressing the terminal fucosylation.
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Background: The objective of this study was to evaluate the serum and salivary L-fucose in oral potentially malignant disorders (OPMDs) and oral cancer (OC) in order to investigate the possibility of using this as biomarker for early diagnosis. Materials and Methods: The study included 85 participants, who were grouped as control (30), OPMDs patients (25), and OC patients (30). Serum and unstimulated whole saliva were collected from participants of all groups and fucose estimation was done using spectrophotometry. The results were tabulated and analyzed statistically. Results: The mean serum L-fucose levels in normal, OPMDs, and OC group were 3.49, 19.18, and 35.75 mg/dl, respectively, while the levels of salivary L-fucose were 3.18, 7.02, and 11.66 mg/dl, respectively. A highly significant rise (P < 0.001) in serum and salivary L-fucose was observed in the study participants compared to control. Conclusions: The present study showed a significant and gradual increase in serum and salivary L-fucose from control to OPMDs to OC. From this study, we suggest that L-fucose can be used as a reliable biomarker and saliva can be used as a diagnostic fluid for screening and early detection of OC
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Background: Cellular glycosylation changes are associatedwith different types of neoplastic transformation. Fucose is adeoxyhexose sugar that the body requires for optimal functionsof cell to cell communications and which plays a role in severalbiological events. Fucose has been considered to play asignificant role in cancer and its spread. Alpha L Fucosidase(ALF) is an exoglycosidase involved in the hydrolyticdegradation of fucose containing components of glycoproteins,glycolipids and oligosaccharides. The significance of thisenzyme in human catabolism is implied by geneticneurovisceral storage disease. Altered levels of ALF has beenreported in the plasma/serum of patients with oral cancer.Aims: To investigate the clinical usefulness of serum fucoseand α-L-fucosidase in diagnosing oral pre-cancer and cancerand study the variations of the levels of both metabolites innormal, precancerous and cancerous conditions (Squamouscell carcinoma).Methodology: The study group comprised of 87 samples of(age range: 20-70 years): control samples – healthy individualswithout any systemic illness (n =20), clinically andhistopathologically diagnosed cases of leukoplakia (n=16) andoral submucous fibrosis (n=16) and oral squamous cellcarcinoma (n=35) respectively. 2ml blood was collected byvenipuncture from every subject after informed consent, serumwas separated and checked for fucose and fucosidase byspectrophotometric analysis.Results: The Normal value range of fucose is 8.3 to 9.5 mg/ dland that of fucosidase is 22.8 ± 7.1 U/L. There is an increasein the value range of fucose and fucosidase in the tissues ofpotentially malignant disorders and Squamous cell Carcinoma.
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Los oligosacáridos de la leche materna (HMOs) son unas 200 moléculas distintas sintetizadas y secretadas por la glándula mamaria a partir de lactosa a la que diversos enzimas unen monosacáridos simples (glucosa, galactosa, n-acetil galactosamina, fucosa y ácido siálico). Estas uniones y sus diferentes orientaciones espaciales generan una gran diversidad de estructuras químicas y de funcionalidades. La concentración de los HMOs es mayor en el calostro (± 25 g/L), está relacionada con la duración del embarazo y la lactancia: disminuyen progresivamente hasta la mitad de los niveles iniciales. La genética materna influye en el perfil de algunos HMOs; el gen FUT2, que codifica la síntesis de la fucosiltransferasa 2 (FUT2) condiciona el llamado carácter secretor en 75-85% de las mujeres y hace que los antígenos del grupo ABO(H) sean secretados en los líquidos orgánicos (saliva, lágrimas, semen). La ausencia de actividad del gen FUT2 condiciona el carácter no-secretor (15-25% de las mujeres). La actividad del gen FUT3 condiciona la actividad de la fucosiltransferasa 3 (FUT3) que se asocia con el grupo sanguíneo Lewis+ mientras que su ausencia caracteriza a los portadores como Lewis 0. Los HMOs son absorbidos a nivel del intestino como trazas (1%) pero incluso en esas cantidades ejercerían efectos sistémicos.
Human milk oligosaccharides (HMOs) are a family of some 200 different molecules synthesized by the mammary gland. At the core is a molecule of lactose, which is linked by different enzymes to glucose, galactose, n-acetyl galactosamine, fucose or sialic acid. These linkages and their different spatial orientation generate, besides the possibilities of numerous chemical structures, the potential for different spatial isomers. The concentration of HMOs in human milk depends on pregnancy and breastfeeding duration. They are highest in colostrum (± 25 g/L) and decrease over time to half this initial level. Maternal genetics modifies the concentration and profile of some oligosaccharides. For example, the FUT2 gene codifies the synthesis of fucosyltransferase 2 (FUT2) whose activity generates the secretor status for antigens of the ABO(H) blood group in organic fluids (saliva, milk, tears, semen) among 75-85% of the carriers of the trait. The absence of activity of the FUT2 gene conditions the non-secretor status (15-25% of women). The FUT3 gene regulates the activity of the fucosyltransferase 3 (FUT3) that is associated with the Lewis blood group. Traces of HMOs (1%) are absorbed in the intestinal tract, however, they exert important systemic effects even at low concentrations.
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Humans , Oligosaccharides , Carbohydrates , Milk, Human , Fucose , LactoseABSTRACT
Objective To investigate the core fucosylated alpha 2-macroglobulin(LCA-α2M) level in patients with hepatocellular carcinoma (HCC), liver cirrhosis (LC) and chronic hepatitis B (CHB), and explore its diagnostic value in HCC. Methods A total of 193 HCC patients,104 LC patients and 71 HC patients in Shanghai Eastern Hepatobiliary Surgery Hospital and 45 CHB patients in Changzheng Hospital from January 2013 to December 2016 were included for retrospective study. The method for detecting LCA-α2M was set up, and then the levels of serum α2M and LCA-α2M in each group were detected. The diagnostic value of LCA-α2M for HCC was evaluated by receiver operating characteristic (ROC) analysis. Result The level of LCA-α2M/α2M × 100(LCA-α2M％) was significantly higher in HCC patients[31.25(26.61-35.42)] than that in LC patients [26.00(22.30-30.64)], CHB patients[26.23 (23.86-31.86)] and healthy controls[20.29(17.35-22.60)] (H values were 5.626, 3.388 and 10.942, respectively, P<0.05). The area under the receiver operating characteristic curve (AUC) of LCA-α2M％for identifying HCC was 0.768 (0.725-0.808). Combined α-fetoprotein(AFP) and LCA-α2M％, the area under the ROC curve was 0.890(0.856-0.919). For AFP negative HCC patients, the sensitivity of LCA-α2M％was 77.42％(24/31). Conclusion LCA-α2M％ has some values in assistant diagnosis of HCC, and could improve the detection of AFP negative HCC patients.
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Glycosylation of the Fc region of IgG has a profound impact on the safety and clinical efficacy of therapeutic antibodies. While the biantennary complex-type oligosaccharide attached to Asn297 of the Fc is essential for antibody effector functions, fucose and outer-arm sugars attached to the core heptasaccharide that generate structural heterogeneity (glycoforms) exhibit unique biological activities. Hence, efficient and quantitative glycan analysis techniques have been increasingly important for the development and quality control of therapeutic antibodies, and glycan profiles of the Fc are recognized as critical quality attributes. In the past decade our understanding of the influence of glycosylation on the structure/function of IgG-Fc has grown rapidly through X-ray crystallographic and nuclear magnetic resonance studies, which provides possibilities for the design of novel antibody therapeutics. Furthermore, the chemoenzymatic glycoengineering approach using endoglycosidase-based glycosynthases may facilitate the development of homogeneous IgG glycoforms with desirable functionality as next-generation therapeutic antibodies. Thus, the Fc glycans are fertile ground for the improvement of the safety, functionality, and efficacy of therapeutic IgG antibodies in the era of precision medicine.
Subject(s)
Animals , Humans , Antibodies, Monoclonal , Pharmacokinetics , Therapeutic Uses , Glycosylation , Immunoglobulin G , Chemistry , Metabolism , Protein Engineering , Methods , Receptors, Fc , Chemistry , Metabolism , Treatment OutcomeABSTRACT
Objective: To establish a polysaccharide fingerprint analysis method for identifying Jinshuibao Capsule. Methods: Polysaccharide from fermentation Cordyceps powder in Jinshuibao Capsule was extracted by boiling water, hydrolyzed by acid, and precolumn derivatization method for 1-phenyl-3-meth yl-5-pyrazolone (PMP), analyzed by HPLC. The chromatographic conditions were Phenomenex OOG-4252-EO C18 column (250 mm × 4.6 mm, 5 μm), eluting with acetonitrile-0.05 mol/L phosphate buffer salt gradient, and the detection wavelength was 250 nm. The column temperature was 30 ℃. Results: Eleven monosaccharide components that made up the fermentation Cordyceps powder were identified and established the fingerprint of fermentation Cordyceps powder polysaccharide. Ten batches of monosaccharide components in Jinshuibao Capsule were analyzed by fingerprints. The similarity was greater than 0.99 with no significant difference. Conclusion: The fingerprints of fermentation Cordyceps powder were simple, specific and reproducible, and the quality of Jinshuibao Capsule is evaluated objectively.
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The present study was aimed to evaluate the antibacterial capability of fucoidan from Sargassum wightii against the chosen human bacterial pathogens. The major chemical constituents of the extracted fucoidan were analyzed by biochemical methods. It showed that the extracted fucoidan contains 52.86 ± 0.64% of fucose and 29.26 ± 0.83% of sulphate. The antibacterial efficacy was performed by agar well diffusion, minimum inhibitory concentration (MIC) and minimum inhibitory concentration (MBC) method. The maximum antibacterial activity 18.6 ± 0.32 mm was obtained for Vibrio cholera and the minimum activity 8.6 ± 0.26 mm was obtained for Salmonella typhi. Result of this manifested the considerable antibacterial potentiality of fucoidan against human bacterial pathogens. Toxicity of fucoidan was evaluated by brine shrimp toxicity assay. No toxic effect was observed in fucoidan. Our study concluded that fucoidan might be used as natural and safe antibiotics in curing many bacterial diseases. Further study is required to get the better understanding of mode of action of fucoidan against the bacterial pathogens.
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The present study was aimed to evaluate the antibacterial capability of fucoidan from Sargassum wightii against the chosen human bacterial pathogens. The major chemical constituents of the extracted fucoidan were analyzed by biochemical methods. It showed that the extracted fucoidan contains 52.86 ± 0.64% of fucose and 29.26 ± 0.83% of sulphate. The antibacterial efficacy was performed by agar well diffusion, minimum inhibitory concentration (MIC) and minimum inhibitory concentration (MBC) method. The maximum antibacterial activity 18.6 ± 0.32 mm was obtained for Vibrio cholera and the minimum activity 8.6 ± 0.26 mm was obtained for Salmonella typhi. Result of this manifested the considerable antibacterial potentiality of fucoidan against human bacterial pathogens. Toxicity of fucoidan was evaluated by brine shrimp toxicity assay. No toxic effect was observed in fucoidan. Our study concluded that fucoidan might be used as natural and safe antibiotics in curing many bacterial diseases. Further study is required to get the better understanding of mode of action of fucoidan against the bacterial pathogens.
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We review the present knowledge of serum fucose with special attention to its relation with various malignant diseases. We summarize the role of serum fucose as a useful diagnostic and prognostic marker when used singly or in combination. The purpose of this review is to provide an expert opinion on the practical and applied aspect of serum fucose level in clinical practice and research settings. Our review is based on information from published research studies, library books, and electronic searches through Medline and PubMed. The available published data were used as the basis for recommendations. Each of the subsections concludes to provide information to assist the clinicians and the research scientists make informed decisions.
Subject(s)
Fucose/blood , Humans , Neoplasms/blood , Prognosis , Biomarkers, Tumor/bloodABSTRACT
S'. rum sialic acid(SA), fucost(Fuc) and five acute phase proteins were measured in 55 patients with digestive tract cancer(DTC), 61 with digestive tract bengin diseases and 58 hcalihy blood donors. The serum concentrations of SA, Fuc, ?1-acid glycoprotcin(?1-AG), ?1-antitrypsin(?1-AT) and haptoglobin (HP) were significantly elevated in the patients with cancer as compared with the control group(P
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Serum concentrations Of sialic acid(SA), fucose (Fuc) , ?1-acid glycoprotein (?1-AG), haptoglobi (HP),?1-antitrypsin (?1-AT), Prealbumin (PA) and tranrf-errin (TF)were co comitantly measured in 31 patier's with AFP negative primary hepatocellular carcinoma (HCC) , 33 with AFP positive HCC, 35 with cirrhosis and 58 normal blood donors. The results revealed that serum SA, Fuc, ?1-AG, HP and ?1-AT levels were all significantly higher in patients with AFP negative HCC than in control groups. Among them, Fuc, ?1-AG and HP levels were significantly higher in patients with AFP negative HCC than in AFP positive HCC. Therefore, measurements of these five indices are suggested in the diagnosis of the AFP negative HCC.Furtheremore, the raised concentration of protein bound sugars in HCC usually bear a strong positive correlation with increased concentrations of serum ?1-AG, HP and ?1-AT
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Serum concentrations of sialic acid, fucose, prealbumin(PA ), ?1-antitrypsin (?1-AT), ?1-acid glycoproteins(?1-AGP), haptoglobin (HP) and transferrin (TF) were measured in 94 nasopharyngeal carcinoma and 65 cancer of larynx. Statistical evalution was made with stepwise discriminant analysis, and the discriminant functions were obtained. The effectiveness of allocation were examined on every one of all patients in the original data, Our results suggested, with the discriminant function equation consisting of serum PA, ?1-AT, ?1-AGP, HP and TF concentrations we could best separate the patients sufferng from nasopharyngeal or laryngeal carcinoma from the normal; the effective rate of discriminant was 87.36% and 88% for nasopharyngeal carcinoma from normal and laryngeal carcin-oma from normal respectively. However using discriminant function equation,composed of serum sialic acid, PA and ?1-AT, we could also discriminate nasopharyngeal or laryngeal carcinoma with recurrence from the one with non-recurrence, the effective rate of discriminant was 93.1% and 80% for nasopharyngeal carcinoma and cancer of larynx respectively