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OBJECTIVE To establish the method for the content determination of 5-hydroxymethylfurfural （5-HMF） in glucosamine hydrochloride tablets， and to analyze its regularity and influential factors. METHODS Quantitative analysis of 5-HMF was performed using high-performance liquid chromatography. The analysis was conducted on Shim-pack GIST C18-AQ column with mobile phase consisted of 0.1% phosphoric acid solution-methanol （90∶10， V/V） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃， and detection wavelength was 284 nm. The injection volume was 20 μL. Reaction kinetics test of different temperatures was adopted to analyze the relationship of 5-HMF content with reaction temperature and reaction time， and utilized to build its formation kinetic model. RESULTS The linger range of 5-HMF was 0.057-5.698 μg/mL （r＝0.999 9）. The limits of detection and quantitation were 5.70 and 17.09 ng/mL； RSDs of precision， repeatability and stability （24 h） tests were all lower than 1.0% （n＝6）. The average recoveries ranged from 99.38% to 99.73%（RSD＝0.53%， n＝9）. The contents of the 5-HMF in 8 batches of samples ranged 4.10-35.13 μg/g. Results of data fitting in reaction kinetics test showed that the higher reaction temperature and the longer reaction time， the higher 5-HMF content in the sample. At 50， 60， 70 and 80 ℃ ， the relationship between the content of 5-HMF and the reaction time was linear， in accordance with a zero-order kinetic model. The reaction rate constants were 6.789， 7.715， 8.815 and 11.430， respectively. CONCLUSIONS The established method has strong specificity， high sensitivity， and good accuracy； the reaction temperature and reaction time are important influential factors for the formation of 5-HMF in glucosamine hydrochloride tables. The change rule of its content conforms to the zero-order kinetic model.
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ABSTRACT: The effect of methods to remove protein content on the properties of glucosamine hydrochloride from the shells of white leg shrimp (Litopenaeus vannamei) and black tiger shrimp (Penaeus monodon) was investigated. Chitin from shrimp shells was obtained by demineralization in 6% HCl for 12h, deproteinization by two different methods (first group soaked in 8% NaOH for 36h and second group treated in Alcalase enzyme at the concentration of 0.2% for 36h). Two group samples were converted to glucosamine hydrochloride by soaking in 36.76% HCl solution for 5h at 85 °C. The results of fourier transform infrared spectroscopy (FTIR), solubility and recovery yield analysis showed that deproteinization methods did not significantly affect the properties of glucosamine hydrochloride. However, glucosamine hydrochloride from white leg shrimp shells contained higher recovery yield and solubility than black tiger shrimp shells.
RESUMO: Investigou-se o efeito de métodos para remover o conteúdo de proteínas nas propriedades do cloridrato de glucosamina das conchas de camarão de pernas brancas (Litopenaeus vannamei) e camarão de tigre preto (Penaeus monodon). A quitina das cascas de camarão foi obtida por desmineralização em HCl a 6% por 12 h, desproteinização por dois métodos diferentes (primeiro grupo embebido em NaOH a 8% por 36 h e segundo grupo tratado na enzima Alcalase na concentração de 0,2% por 36 h). Duas amostras de grupo foram convertidas em cloridrato de glucosamina por imersão em solução de 12M HCl por 5 h a 85 °C. Os resultados das análises de FTIR, solubilidade e rendimento de recuperação mostraram que os métodos de desproteinização não afetaram significativamente as propriedades do cloridrato de glucosamina. No entanto, o cloridrato de glucosamina de cascas de camarão de pernas brancas continha maior rendimento e solubilidade de recuperação do que as cascas de camarão tigre preto.
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Abstract The objective of the present study is to develop and validate a simple, selective and accurate hydrophilic interaction liquid chromatography - a high performance liquid chromatography incorporating an evaporative light scattering detector (HILIC-HPLC-ELSD) method for simultaneously determining glucosamine hydrochloride and chondroitin sulfate in dietary supplements. The chromatographic separation was carried out on a ZIC-HILIC column (150 mm x 4.6 mm x 5µm) in isocratic system mode with a mobile phase of acetonitrile, 30 mM ammonium formate and water (77:20:3, v/v/v) at pH 4.5, a column temperature of 35°C, a flow rate of 1 mL.min-1, and an injection volume of 5 µL. An evaporative light scattering (ELS) detector was used. Effective separation was achieved by means of analyte resolution of more than 1.5 with an analysis run time of approximately 20 minutes. The linearity of glucosamine hydrochloride and chondroitin sulfate ranged from 0.4 to 2.5 mg.mL-1. The limits of the detection and quantification of glucosamine hydrochloride were 20 and 80 mg.mL-1 respectively, while for chondroitin sulfate they were 80 and 400 mg.mL-1. All validation parameters satisfied the acceptance criteria in accordance with International Conference on Harmonisation (ICH) guidelines. The method was successfully applied to the assay of commercial dietary supplement samples
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Chromatography, High Pressure Liquid/methods , Chromatography, Liquid/methods , Dietary Supplements/analysis , Validation Study , Glucosamine/agonistsABSTRACT
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.
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Objective: To study the effect of Bushen Zhuangjintang and glucosamine hydrochloride on the repairing of rat knee cartilage defect, the combined application of traditional Chinese and western medicine can promote the repairing of knee cartilage defect more effectively. To provide a new theoretical basis and method for the treatment of knee cartilage injury. Method: SPF rats (64 rats) were randomly divided into 5 groups, sham group,model group, Bushen Zhuangjintang group (7.5 g·kg-1·d-1), and glucosamine hydrochloride group(7.5 g·kg-1·d-1), Bushen Zhuangjin Decoction combined with glucosamine hydrochloride group[(7.5+7.5) g·kg-1·d-1] was administered to rats by drug gavage. The knee cartilage defects were repaired by gross observation and scanning electron microscopy at 4, 8 and 12 weeks. Real-time PCR was used to detect the expression of Collagen Ⅱ and Aggrecan mRNA in each group. Western blot was used to detect the expression of Collagen Ⅱ. Result: 4,8,12 weeks compared with group, Bushen Zhuangjintang combined with hyaluronic acid in the glucosamine hydrochloride group was filled with hyaline cartilage. The cartilage in the drilled area was smooth and smooth, and integrated with the surrounding cartilage tissue, which was superior to other groups. Real\|time PCR and Western blot analysis showed that the expression of Aggrecan protein and mRNA were significantly increased at 4, 8 and 12 weeks compared with the blank group (PPConclusion: The combined application of traditional Chinese and western medicine can obviously promote the repair of cartilage defects in the knee of rats. The possible mechanism of the treatment of cartilage injury by the combination of traditional Chinese and western medicine is analyzed from the level of gene protein expression and microstructure.
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Aim: To determine the pharmacokinetics and bioavailability of glucosamine after administration of glucosamine hydrochloride (GH) and glucosamine sulfate (GS). Methods: GH or GS was given ig and iv to rats, respectively. An LC-MS/MS method was developed to measure the concentrations of glucosamine in plasma. DAS 3. 0 was employed to obtain the pharmacokinetical parameters as well as bioavailability. Results: There was no marked difference in T1/2, Vz and absolute availability between GH and GS. However, the relative bioavailability of GH was higher than that of GS. In addition, Tmax of GS was shorter than that of GH. Conclusion: There are some differences between the glucosamine pharmacokinetics of GH and GS.
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BACKGROUND: Glucosamine hydrochloride (GlcN·HCl) has been shown to inhibit cell growth and matrix synthesis, but not with N-acetyl-glucosamine (GlcNAc) supplementation. This effect might be related to an inhibition of critical growth factors (GF), or to a different metabolization of the two glucosamine derivatives. The aim of the present study was to evaluate the synergy between GlcN·HCl, GlcNAc, and GF on proliferation and cartilage matrix synthesis. METHOD: Bovine chondrocytes were cultivated in monolayers for 48 h and in three-dimensional (3D) chitosan scaffolds for 30 days in perfusion bioreactors. Serum-free (SF) medium was supplemented with either growth factors (GF) TGF-β (5 ng mL₋₁) and IGF-I (10 ng mL₋₁), GlcN·HCl or GlcNAc at 1mM each or both. Six groups were compared according to medium supplementation: (a) SF control; (b) SF + GlcN·HCl; (c) SF + GlcNAc; (d) SF + GF; (e) SF + GF + GlcN·HCl; and (f) SF + GF + GlcNAc. Cell proliferation, proteoglycan, collagen I (COL1), and collagen II (COL2) synthesis were evaluated. RESULTS: The two glucosamines showed opposite effects in monolayer culture: GlcN·HCl significantly reduced proliferation and GlcNAc significantly augmented cellular metabolism. In the 30 days 3D culture, the GlcN·HCl added to GF stimulated cell proliferation more than when compared to GF only, but the proteoglycan synthesis was smaller than GF. However, GlcNAc added to GF improved the cell proliferation and proteoglycan synthesis more than when compared to GF and GF/GlcN·HCl. The synthesis of COL1 and COL2 was observed in all groups containing GF. CONCLUSION: GlcN·HCl and GlcNAc increased cell growth and stimulated COL2 synthesis in long-time 3D culture. However, only GlcNAc added to GF improved proteoglycan synthesis.
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Bioreactors , Cartilage , Cell Proliferation , Chitosan , Chondrocytes , Collagen , Glucosamine , Insulin-Like Growth Factor I , Intercellular Signaling Peptides and Proteins , Metabolism , Methods , Perfusion , ProteoglycansABSTRACT
Objective To study the clinical effect of psychological intervention combined with oral administration of aminoglucose hydrochloride capsules combined with intra-articular injection of sodium hyaluronate combined with functional exercise in the treatment of knee osteoarthritis.Methods Forty-eight patients with knee osteoarthritis were randomLy divided into control group and study group(n=39).The patients were randomLy divided into control group and study group.The control group was given intraperitoneal injection of sodium hyaluronate combined with functional exercise, the study group on the basis of the control group to give psychological intervention and oral administration of aminoglucose capsules combined therapy.Comparison of two groups of Lysholm knee function score and postoperative efficacy.Results The scores of Lysholm knee function were significantly improved after treatment, but the improvement rate of the study group was significantly better than that of the control group (P<0.05).The clinical effect of the study group was significantly better than that of the control group(94.87% vs 43.59%).The difference was statistically significant (P<0.05).Conclusion The combination of psychological intervention combined with oral administration of aminoglucose capsules with intraperitoneal injection of sodium hyaluronate combined with functional exercise in the treatment of knee osteoarthritis can significantly improve the clinical efficacy, improve the knee function and improve the prognosis, worthy of clinical application and promotion.
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OBJECTIVE: To establish an HPLC method for determination of the related substances and content of glucosamine hydrochloride products from different manufacturers, and evaluate the quality variation of products of three dosage forms from eight different manufacturers. METHODS: The determination method was developed on a Phenomenex Luna NH2 column(4.6 mm × 250 mm, 5 μm), with phosphate buffer (7.0 g dipotassium hydrogen phosphate dissolved in a suitable amount of water, diluted with 0.5 mL ammonia, add water to 2 000 mL, adjusted to pH 7.5 with phosphoric acid)-acetonitrile (25:75) as mobile phase at a flow rate of 1.5 mL· min-1. The column temperature was maintained at 30℃ and detection wavelength was set at 195 nm. RESULTS: The system suitability test met the requirements, and the resolution between the peaks of glucosamine hydrochloride and the impurities were greater than 1.5. Good linearity was shown for glucosamine hydrochloride in the concentration range of 0.156 64-15.664 mg·mL-1 (r=0.999 9). The average recovery rate was 101.14%, with RSD of 1.20% (n=9). CONCLUSION: The established method can be applied in the simultaneous determination of the related substances and content of glucosamine hydrochloride products and provide reference for the quality control.
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Objective:To establish an HPLC method for the determination of indomethacin and glucosamine hydrochloride in glu-cosomine and indomethacin enteric-coated tablets. Methods:A waters Bridge C18(250 mm×4.6 mm,5 μm) column was used with ammonium dihydrogen phosphate (the pH value was adjusted to 3.0 by phosphoric acid) (A) and acetonitrile (B) as the mobile phase with gradient elution,the flow rate was 0.7 ml·min-1,the detection wavelength was 195 nm,the sample volume was 20 μl and the column temperature was 30℃. Results: Glucosamine hydrochloride and indomethacin had a good linear relationship within the range of 0.030 0-1.500 8 mg·ml-1(r=0.999 1) and 0.010 3-0.513 0 mg·ml-1(r=0.999 9), and the average recovery was 98.3%(RSD=0.33%,n=6)and 99.9%(RSD=0.06%,n=6), respectively. Conclusion: The method is accurate, rapid, sensi-tive and specific,and can be used to control the quality of the product.
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OBJECTIVE: To establish a1 H-NMR internal standard method for measurement of glucosamine hydrochloride in glucosamine hydrochloride capsules. METHODS: For the HPLC-UV method, a China national drug tentative standard (No. YBH11442006) was employed. For the H-NMR analysis, the optimal determination conditions were selected as follows; the pulse sequence was zgpr, hydrogen spectral width was 6 009 Hz, pulse width was 11. 95μs, delay time was 6.50μs, the temperature was 27℃ and the number of scans was 32. RESULTS: The peak of δ 3.05 ppm of the glucosamine hydrochloride and the peak of δ 0 ppm of TSP were selected as quantitative peak and internal standard peak, respectively, and the concentration of the internal standard TSP was 1.02 mmol·L-1. A good linear relationship was obtained with a regression equation as Y = 2.717 IX +0.149 9(r =0.9998) in the concentration range of 2.0-12.0 mg·mL-1. The average recovery rate and the RSD of glucosamine hydrochloride capsules were 103.59% and 1.09% (n = 9), respectively. CONCLUSION: With simple pretreatment, high sensitivity, good repeatability and short analysis time, the NMR-based method could be applied to the measurement of glucosamine hydrochloride in glucosamine hydrochloride capsules.
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Objective To establish an high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method for determination of glucosamine hydrochloride and its related substances in glucosamine and indometacin enteric-coated capsules.Methods Separation was carried out on a Inertsil C8 Column (250 mm ×4.6 mm,5μm) with a mobile phase composed of acetonitrile-water(70∶30) at a flow rate of 1.0 mL/min and the column temperature was 25℃.Results The linear ranges of calibration curve was 25 -312.5 μg/mL(r=0.9991, n=6).The average recovery was 98.0%, and the relative standard deviation was 1.3%( n =5 ) .The glucosamine hydrochloride was stable with acid destruction, but unstable with alkali, heat and strong light destruction.Conclusion The method is simple, accurate and reproducible for determination of glucosamine hydrochloride and related substances.
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Objective: To establish an HPLC method for determining the content of glucosamine hydrochloride and its prepara-tions. Methods:A Phenomenex Luna NH2 column was used, and the mobile phase was ammonium phosphate buffer (dissolving 3. 5 g dipotassium phosphate and 0. 25 ml ammonium hydroxide into 1 000 ml water, adjusting pH to 7. 5 with phosphoric acid) – acetoni-trile (25 ∶75). The flow rate was 1. 5 ml·min-1 and detection wavelength was 195 nm. The column temperature was 35℃ and the injection volume was 20 μl. Results:The linear range of glucosamine hydrochloride was 1. 6-16. 0 mg·ml-1(r=0. 999 9). The av-erage recovery of glucosamine hydrochloride tablets and capsules was 99. 3% and 99. 5%, respectively, and RSDs were 0. 3% ( n=9). Conclusion:The method is simple, accurate and reliable, and suitable for the quality control of glucosamine hydrochloride and its preparations.
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OBJECTIVE: To discuss the effect of glucosamine-hydrochloride (Glu/Ch) in protecting and repairing the cartilage in blood-induced joint damage (BJD) in vivo. METHODS: Thirty-two adult New Zealand rabbits were randomly divided into 4 groups (n=8):high-dose Glu/Ch treated group (group A), low-dose Glu/Ch treated group (group B), positive control group (group C), and negative control group (group D). A joint bleeding model was established by blood injection into articular cavity in groups A, B, and C. Glu/Ch was given by gavage in groups A (250 mg/kg) and B (21.5 mg/kg) once a day for 8 weeks, and the same dosage of saline was given in groups C and D. The serum cartilage oligomeric matrix protein (COMP), serum chondroitin sulfate 846(CS846), and urinary C-terminal telopepide of type II collagen (CTX-II) were measured at 3 days, 7 days, 2 weeks, and 8 weeks after modeling. The expressions of cytokines such as interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) in synovial fluid were analyzed by ELISA at 8 weeks after modeling. The expression of matrix metalloproteinase 13(MMP-13) was detected by immunohistochemistry. Alcian blue staining and Safranin-O staining were performed to calculate the percentage of the positive staining areas. The proteoglycan content was detected by semi-quantitative analysis in the articular cartilage. RESULTS: The COMP concentration was significantly higher in groups A, B, and C than group D, and in groups B and C than group A at 3 days after modeling (P0.05), and it was significantly lower in groups A, B, and D than group C (P0.05). Difference in CS846 concentration had no significance among 4 groups at each time point (P>0.05). The CTX-II concentration of groups A, B, and C was significantly higher than that of group D at each time point (P0.05). The IL-1β concentration was significantly higher in group C than the other groups (P0.05). The MMP-13 expression was significantly higher in group C than groups A, B, and D (P<0.05), in groups A and B than group D (P<0.05). A significant decrease in the area stained with Alcian blue and Safranin-O was observed in group C. There were significant differences in the percentage of the positive stained areas of Alcian blue and Safranin-O among 4 groups (P<0.05). The relative quantities of proteoglycan from small to large in order was groups C, B, A, and D, respectively, showing significant differences (P<0.05). CONCLUSIONS: The metabolism disorder of cartilage matrix and synovium inflammatory reaction can be observed in rat joint bleeding model. Glu/Ch has certain protective effect on the cartilage after BJD by down-regulating IL-1β, TNF-α, and MMP-13, as well as increasing proteoglycan content in the cartilage.
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Objective To explore the clinical value of Glucosamine hydrochloride tablets(GH)alone and combination with Xianlinggubao capsules(XLGB)for treatment patients with knee osteoarthritis(KOA).Methods 96 patients with KOA were selected,and were divided into two groups according to the random number method, 48 cases in each group.The control group was treated with GH alone,while the observation group was treated with XL-GB on the basis of the control group.US Western Ontario and McMaster University Osteoarthritis Index (WOMAC) were used to evaluate the symptoms change,including pain,stiffness,physical function scores and total scores before and after treatment.The onset time,knee function (HSS)score at 3 months and 6 months after treatment were recor-ded.CRP and ESR were measured,and the total efficacy was evaluated at the end of treatment.Results The differ-ence of WOMAC score between the two groups was not statistically significant before treatment (P >0.05 ).After treatment,pain,stiffness,physical function scores and total scores were (10.2 ±1.4)points,(3.5 ±1.6)points, (40.5 ±5.0)points and (56.4 ±6.7)points in the observation group,those were (14.8 ±2.6)points,(6.2 ±2.3)points, (52.2 ±6.8)points and (73.3 ±4.5)points in the control group,the differences were statistically significant (t =7.631,4.721,6.791,10.26,all P <0.01).The onset time,HSS score at 3 months and 6 months after treatment in the observation group were (6.0 ±2.2)d,(68.2 ±6.4)points and (84.3 ±6.2)points,while those were (9.8 ± 2.8)d,(58.5 ±3.9)points and (72.8 ±5.4)points in the control group,the differences were statistically significant (t =5.228,6.341,6.852,all P <0.01 ).The improvements of CRP level and ESR in the observation group were more pronounced than those in the control group (t =3.880,2.668,all P <0.05).After treatment,the total effective rate of the observation group was 93.8%(45 /48),while that was 77.1%(37 /48)in the control group,the difference was statistically significant (χ2 =5.352,P <0.05).Conclusion GH combines with XLGB can significantly improve symptoms in patients with KOA,with more rapid recovery of knee function,thus it is a safe and effective therapy.
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ObjectiveTo investigate the effect of combined therapy with external use of wishful golden cream and oral use of glucosamine hydrochloride on knee osteoarthritis.Methods200 patients with knee osteoarthritis were divided into treatment group and control group according to the order of treatment.Control group were given one piece of oral glucosamine at a time,three times a day.It consisted of six weeks treatment for lcowrse,two courses per year.Treatment group were given oral glucosamine plus external usage of wishful golden cream every other day with a new one.It consisted of 14 days for a course,two courses per year.ResultsAll patients were followed up for 1year.The efficien rate of the treatment group was 85.0％,and significantly higher than than of the control group (73.0％) ( x2 =4.34,P ＜ 0.05 ).ConclusionCombined therapy with wishful golden cream and oral usage of glucosamine hydrochloride on knee osteoarthritis could significantly improve the clinical symptoms.It was more effective than oral treatment with glucosamine hydrochloride alone and it had great clinical value.
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Objective To study the catalytic capability of glucosamirie-Cu( Ⅱ) complex for decomposition of H2O2 and its relative factors. Methods Glucosamine-Cu( Ⅱ ) complex was prepared by the reaction of D-glucosamine hydrochloride with Cu2+ in aqueous solution, then added it into H2O2 solution. The concentration of H2O2 was determined by titrimetric analysis in a regular interval of time, the rate of decomposition of H2O2 was obtained in various conditions. Results Strong catalytic capability of glucosamine-Cu( Ⅱ ) complex was obtained at 30℃ pH 6. 5, the rate of decomposition was over 90% after 12h, and was almost 100% after 24h. Conclusion The complex of glucosamine-Cu( Ⅱ ) showed good catalytic capability for decomposition of H2O2.
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Objective To study the effects of D-glucosamine hydrochloride on the apoptosis of gastric carcinoma cell line SGC-7901 and its mechanism.Methods SRB assay was used to examine the effects of D-glucosamine hydrochloride on the growth inhibition of SGC-7901.The cell-cycle and expression of cytochrome-C were observed by flow cytometry,the concentrations of intracellular Ca2+ was determined by CLSM and the expressions of cyclinB1,calcinerin were investigated by Western-Blot.Results D-glucosamine hydrochloride could block SGC-7901 cell-cycle at S phase of cell division,increase the concentration of intracellular Ca2+ and decrease the expression of cyclinB1,the expressions of calcinerin and cytochrome-c were increased respectively.Conclusion Block of cell-cycle may be one of the mechanism of inducing cell apoptosis by D-glucosamine hydrochloride.
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Objective To study the effect of GAH on lymphocytes proliferation and its mechanism. Methods MTT, fluorescence probe and immunofluorescent methods were used to investigate the effects of GAH on the lymphocytes proliferation, intracelluar Ca2+ concentration and expression of calcineurin(CaN). Results GHA induced lymphocyte proliferation, raised the intracelluar Ca2+ concentration and calcineurin expression. Conclusions GAH is a new kind of activator, which can induce lymphocyte proliferation through Ca2+ signal pathway.
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Objective The growth inhibitory effects of GlcNH_2?HCl,GlcNH_2 and NAG on human hepatoma SMMC-7721 cells in vitro was investigated.The antitumor activity of GlcNH_2?HCl against Sarcoma_(180) in KM mice was also investigated.Methods The cell viability in vitro was examined with MTT assay.DNA isolation and cell-cycle analyses were also performed.GlcNH_2?HCl was ig administered to Sarcoma_(180) KM mice.The inhibition rates,spleen and thymus index were calculated.Results GlcNH_2?HCl and GlcNH_2 resulted in a concentration-dependent reduction in hepatoma cell growth.The inhibition rates against SMMC-7721 cell of GlcNH_2?HCl and GlcNH_2 at concentration of 500?g?mL~(-1) were(50.24)% and 52.19%.As to the concentration of 1000?g?mL~(1), the rates were(82.21%) and 83.20%.This effect was accompanied by a marked increase in the proportion of S phase cells.Compared with the control,there was no significant difference among various concentrations of NAG,GlcNH_(2)?HCl exhibited inhibitiory effect against Sarcoma_(180) in mice at the dosage of 125～500 mg?kg~(-1),and the inhibition rate was about 27.84%～34.02%.The optimal inhibitory effect was 250 mg?kg~(-1).GlcNH_2?HCl could enhance the weights of thymus and spleen.In addition,it could promote lymphocyte transformation.Conclusion It is therefore postulated that the antitumor effect of GlcNH_2?HCl is probably host-mediated and cytocidal.