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Objective To investigate the protection and mechanism of procyanidins (PC) against H2O2 induced oxidative damage of human trabecular meshwork cells (HTMC) in order to provide an experimental foundation for glaucoma clinical treatment.Methods HTMC were cultured and then divided randomly into 5 groups.As untreated group:Normal cultured HTMC;Control group:Normal cultured HTMC + H2O2 (500 μmol · L-1 for 1 hour);Treated group:Normal cultured HTMC + H2O2 (500 μmol ·L-1 for 1 hour) + PC (PC fmal concentrations were 0.02 g · L-1,0.05 g · L-1,0.10 g· L-1).Real-time fluorescence quantitative polymerase chain reaction (PCR) was used to investigate the expression of mitochondrial complex Ⅰ mRNA.Results Compared with untreated group (1.000 0 ± 0.000 0),the differences of mitochondrial complexⅠ mRNA expression in 0.02 g · L-1 PC (0.401 3 ±0.010 3),0.05 g · L-1 PC (0.791 5 ± 0.008 5) groups were statistically significant (all P ＜ 0.01),but the 0.10 g ·L-1 PC group (1.043 0 ± 0.062 2) had no significant differences (P ＞ 0.05).The differences between PC treated groups and control group were statistically significant (P ＜0.01),which showed HTMC treated with PC could increase the expression of mitochondrial complex Ⅰ mRNA.The differences in each PC treated groups were statistically significant (P ＜ 0.01),which showed the expression of mitochondrial complex Ⅰ mRNA were increased along with the concentration of PC gradually increased.Conclusion Exogenetic PC can increase the expression of mitochondrial complex Ⅰ mRNA in the oxidative damaged HTMC,and in a certain range of concentration,the protective effects of PC have the positive relationship of dose-effect,which suggest that PC may be a good candidate for further study of the clinical treatment of glaucoma.
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AIM To study the chemical constituents from Phyllodium pulchellum (L.) Desv..METHODS The ethyl acetate and n-butanol fractions of P.pulchellum 95％ ethanol extract were isolated and purified by Sephadex LH-20,silica,ODS and pre-HPLC column,then the structures of isolated compounds were identified by physicochemical properties and spectral data.RESULTS Eleven compounds were isolated and identified as phydroxybenzoic acid (1),protocatechuic acid (2),protocaechuic acid methyl ester (3),protocatechuic acid ethyl ester (4),ethyl gallate (5),p-coumaric acid (6),caffeic acid ester (7),loliolide (8),uridine (9),arbutin (10),daucosterol (11).CONCLUSION Compounds 4-11 are isolated from genus Phyllodium for the first time,compounds 2 and 3 are first obtained from this plant.
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Marine traditional Chinese medicine (MTCM) is an important part of Chinese medicine (CM),there are some differences in understanding of the current literature and the extension of the connotation of marine medicine,which leads to the definition dispute of MTCM,hindering clinical application and further development of MTCM.In this study,we explored the concept of MTCM in literature,discussed the attributes of ocean marine CM,summed up the differences between the land CM and MTCM over variety characteristics,effect of drug composition characteristics and biological activity characteristics,and discussed the connotation and extension of MTCM from three aspects of theoretical basis and the effect and source of the drug,leading to the formation of the narrow and broad concept of MTCM.The five kinds of disputes in the definition of MTCM were discriminated according to the concept and connotation,which provided a theoretical basis for the definition and the research of MTCM.Moreover,we also defined the English translation and its abbreviation as Marine Traditional Chinese Medicine (MTCM).
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Marine Chinese Medicine (MCM) is one of the important part of the traditional Chinese medicine.The exploration of marine organism resources provide a good base for the development of MCM.However,the evaluation for the nature of the new source of MCM becomes one of the key problem of the clinic application of MCM.In this study,613 MCM and their related 1 091 species of marine organisms were screened.Association Rules Mining method and Phylogenetic Tree constructing method were used to find out the association relationship and the distribution regularity of the MCM with different nature in the Phylogenetic Tree.The results showed high collection of the MCM with the same nature on the tree.The MCM from the organisms in the same family might have the same nature.And the association rules for the same nature assembled at the same branch or near branch of the tree.For example,the marine plantae,Chlorophyta,Florideophyceae,and Phaeohpyceae,were related with cold nature,while the marine animalia,including Decapoda,Malacostraca,and Arthropoda,had close relationship with hot nature.Moreover,the neutral nature was related with Squamata.These results implied that there were close relationship between the nature and the affinity relationship.The MCM from the same or close families may have the same or likely nature.This study provided a new index and reference for prediction and evaluation of the nature of new MCM.
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The quality of traditional Chinese medicines (TCMs) has been mainly evaluated based on chemical ingredients, yet recently more attentions have been paid on biological ingredients, especially for pill-based preparations. It is a key approach to establish a fast, accurate and systematic method of biological ingredient analysis for realization of modernization, industrialization and internationalization of TCMs. The biological ingredient analysis of TCM preparations could be abstracted as the identification of multiple species from a biological mixture. The metagenomic approach based on high-throughput-sequencing (HTS) and big-data-mining has been considered as one of the most effective methods for multiple species analysis of a biological mixture, which would also be helpful for the analysis of biological ingredients in TCMs. Simultaneous identification of diverse species, including the prescribed species, adulterants, toxic species, protected species and even the biological impurities introduced through production process, could be achieved by selecting appropriate DNA biomarkers, as well as applying large-scale sequence comparison and data mining. By this approach, it is prospective to offer an evaluation basis for the effectiveness, safety and legality of TCM preparations.
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Experience mode of marine medicine (EMMM) is the summary of experiences on marine medicine con-cluded by ancient doctors for thousands of years. The mining of EMMM is very important for the clinic application and development of marine medicine. However, these experiences are very difficult to discover for they are all scat-tered in the large number of ancient documents. We proposed that the EMMM can be built by the procedure of in-formation of ancient documents, text-mining of ancient documents, and associate network of informatics unit of ma-rine medicine by non-interactive literature-based knowledge discovery methods, and expert evaluation at the end, so as to provide database for the clinic application and further development of marine medicine.
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ated acute peritonitis induced by LPS in rats.
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Objective To explore the effects of peroxisome proliferator-activated receptorγ (PPARγ) agonist rosiglitazone and 15-deoxy-delta-12,14-prostaglandin J2 (15d-PGJ2) on the expression of PPARγ, toll-like receptor 4 (TLR4) and the activation of STAT1 as well as the local inflammation reaction of abdominal cavity in sprague dawley (SD) rats with peritoneal dialysis- related acute peritonitis induced by lipopolysaccharide (LPS). Methods Twenty-four male SD rats were equally randomized to four groups(n=6 each): control group, injected with 4.25% dextrose peritoneal dialysate (PDF) via abdominal cavity(90 ml/kg); LPS group, injected with LPS(1 mg/kg) via abdominal cavity 4 hours later follewed by PDF injection; rosiglitazone plus LPS group (Rosi group), preconditioned with rosiglitazone (20 mg·kg-1·d-1) by intragastric way for 3 days, then injected with LPS and PDF via abdominal cavity; 15d-PGJ2 plus LPS group (15d-PGJ2 group), preconditioned with 15d-PGJ2 (0.3 mg·kg-1·d-1)via abdominal cavity injection for 3 days, then injected with LPS and PDF via abdominal cavity. The rats were killed 4 hours after PDF injection, IL-6 level in abdominal dropsy was determined by ELISA. Peritoneum tissue was stained by Masson. Leucocyte count in abdominal dropsy was performed. The mRNA expression of PPARγ and TLR4 in peritoneum tissue was determined by RT-PCR; the protein expression of PPARγ, TLR4, p-STAT1 and STAT1 in peritoneum tissue was analyzed by Western blot. Results IL-6 level of abdominal dropsy in LPS group [median 268.53 (range 201.87-335.19) ng/L] was significantly higher than that of control group [median 147.62 (range 130.60-164.64) ng/L] (P<0.01). The IL-6 level of abdominal dropsy in Rosi group [median 110.20 (range 77.60-142.80) ng/L] was significantly lower than that of LPS group (P<0.05). Compared to that of control group, the edematous degree of peritoneum in LPS group was significantly severer, meanwhile, mRNA and proteins expression of PPARγ and TLR4 in rat peritoneum were also significantly higher (P<0.05, P<0.01). Compared to that of LPS group, the edematous degree of peritoneum in Rosi group was lighter, the expression of PPARγ and TLR4 mRNA was significantly up-regulated (P<0.05), meanwhile their proteins expression was down-regulated (P<0.05); and in 15d-PGJ2 group, the edematous degree of peritoneum, the expression of PPARγ mRNA and protein was also decreased (P<0.05), but TLR4 mRNA expression was up-regulated (P<0.01), however, its protein expression was down-regulated (P<0.05). There were no significant differences in leucocyte count of abdominal dropsy among the four groups. The p-STAT1 expression in the rats peritoneum induced by LPS was markedly increased by both rosiglitazone and 15d-PGJ2 (P<0.01). Conclusions Both rosiglitazone and 15d-PGJ2 can down-regnlate the inflammatory reaction in rat peritonitis induced by LIPS, which may be involved in modulating the expression of associated functional protein during LPS signal pathway.
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Purpose To produce enzymatic hydolysates with angiotensin Ⅰ-converting enzyme (ACE) inhibitory activity and antihypertensive activity from Acetes chinensis. Methods ACE inhibitory activity of the hydrolysates of Acetes chinensis by five commercial proteases were determined in vitro to select a protease as the enzyme used in the preparation of enzymatic hydrolysate of Acetes chinensis, and orthogonal trials were employed to optimize its hydrolysis parameters, spontaneously hypertensive rats (SHR) were used to assess in vivo the hypotensive effects of hydrolysate under optimized condition. Results The hydrolysate with IC50 being0.65 mg/ml was obtained under the optimized condition of pH 2.4, 41℃, 3 h hydrolysis time, 3% enzyme/substrate ratio (E/S) and 8 % substrate concentration, and a recorded SBP reduction of 3 886.3 Pa (29mmHg) at 4 h after administration (1.0 g/kg BW) was observed. Conclusion Peptic hydrolysate of Acetes chinensis shows significant ACE inhibitory activity and antihypertensive activity.
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Objective To investigate the expression of E26 transformation-specific-1 (E26ts-1),matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in choroidal melanoma and the correlation with the tumor′s infiltration and metastasis. Methods Immunohistochemistry was used to detect the expression of E26ts-1,MMP-1and TIMP-1 in 78 cases of choroidal melanoma who were divided into shuttle-cells,paraepithelial-cells and mixed-cells type according to the configuration of tumor cells.The patients were followed up and their average existing time was calculated.The results were statistically computed with statistic SPSS 10.0 package. Results In the 78 cases,shuttle-cells type was found in 21,paraepithelial-cells type in 34,and mixed-cells type in 23. Expression of TIMP-1was low in uveal melanoma,while expression of E26ts-1 and MMP-1 was obviously found in the three types of choroidal melanoma;the sequence of expression intensity was shuttle-cells,mixed-cells and paraepithelial-cells type.Among 37 cases who had been followed up,the shuttle-cells type was in 18 with the average existing time of (78.33?24.69) months,the mixed-cells type was in 10 with the average existing time of (61.44?20.46) months,and the paraepithelial-cells type was in 9 with the average existing time of (36.76?12.19) months.The existing time was negative correlated with the intensity of expresion of E26ts-1 and MMP-1. Conclusion The high expression of E26ts-1 and MMP-1and low expression of TIMP-1may relate to the choroidal melanoma′s infiltration and metastasis.
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AIM:To study the effects and mechanism of recombinant human defensin ?1 on cell proliferation in cultured rat glomerular mesangial cells.METHODS:The influences of defensin ?1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay.The different concentrations of U0126,signal-regulated protein kinase(MEK)inhibitor,were added into the culture mediums of mesangial cells to do blocking test.Incubated with a final concentration of 3 mg/L defensin ?1,the phosphorylation of extracellular signal regulated kinase(ERK)1/2 and type IV collagen of mesangial cells in different times were evaluated by Western blotting.RESULTS:Defensin ?1 at 3-20 mg/L enhanced proliferation of rat glomerular mesangial cells.The incubation times for the maximum effect on proliferation was 12 h(P20 mg/L decreased cell proliferation.The cell proliferation induced by defensin ?1 was inhibited by U0126.Stimulation of the cells with defensin ?1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK.After 12 h incubation with defensin ?1,an increase in type IV collagen was observed by Western blotting and continued to increase at 24 h and 48 h(P
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Glycoprotein of Chlamys ( Azumapecten ) farreril ( GCFI) was prepared from the skirt of scallop Chlamys (Azumapecten)farreri by extraction with hot water,isolation and purification with column chromatography "on DEAE-cellulose and Sephadex G-100 successively. GCFI was hydrolyzes with enzymes to obtain glycoprotein of Chlamys (Azumapecten)farreri IKGCFID-The chemical composition and antitumor activity of GCFI and GCFII were studied. The result showed that GCFI and GCFII were two glycoproteins in which the composition and content of protein and sugar were different. They could obviously inhibit Sarcoma 180 cells proliferation of mice with the inhibition rate of 47. 29% and 46. 97%,respectively. Pharmacological test indicated that antitumor activity of GCFI and GCFII were associated with the structure of sugar in the molecules.
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The glycosaminoglycan (GAG), isolated from Bay scallop Argopectenirradians, contains neutral monosaccharides besides hexosamines and hexosuronic acids. The monosaccharides obtained by alcoholysis with HCI-methylalcohol from the sample of GAG was trimethylsilanized with hexamethyldisilan and chlortrimethylsilan (HMDS ' TMCS = 2 : 1). And the trimethylsilyl derivatives of monosaccharides was determined by gas chro-matography. Compared the gas chromatography of the sample with that of standard monosaccharides, it was found that the GAG of the Bay scallop contains five neutral monosaccharides, viz glucose, galactose, xylose, fucose and rhamnose.
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Fraction F3-3-3 of glycosaminoglycan was isolated from the leftover bits of Bay scallop Argopecten irradiaus . Infrared spectra of F3-3-3 was similar to that of heparin. Agarose gel electrophoresis of this fraction was performed in the system of barbital buffer (0. 06 mol/L,pH 8. 6). It was found that F3-3-3 showed single band in the electrophoresis. Compared with the agarose gel electrophoresis of standard glycosaminoglycan, such as heparin, hyaluronic acid and chondroitin 6-sulfate, it indicated that the relative migration rate of the fraction was close to that of heparin.
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The studies on glycosaminogiycans (GAG) from animals are carrying out extensively in the fields of biology.biochemistry and medicine. The methodology of GAG researches has been developed rapidly at present. The researches of GAG have just begun in China. Synthesizing a large number of references,this paper gives a brief introduction on the methods of extraction isolation and determination for GAG. And the latest methods and developing tendency are summarized as well.
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Objective To investigate the chemical constituents of gorgonian Muriceides collaris. Methods The compounds were isolated and purified by silica gel column,Sephadex LH-20 column chromatography and HPLC. Their chemical structures were identified by physicochemical analysis,spectroscopic analysis,and comparison with the data of literatures. Results From the CH3OH soluble extract of Muriceides collaris,nine compounds were isolated,and their structures were determined as cholesterol (1),batyl alcohol (2),benzoic acid (3),uracil (4),thymine (5),2'-deoxyuridine (6),2'-deoxythymidine (7),thymidine (8) and 2'-deoxyadenosine (9),respectively. Conclusion All compounds were obtained from the species Muriceides collaris for the first time.
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Objective To report a method for preparation of saturated oligosaccharides from al- ginate. Methods The alginate was degraded by acid hydrolysis, followed by fractionation at pH 2.85 resulting in the homopolyguluronic acids (PG) and homopolymannuronic acids (PM). The alginate-derived homopolymers were further degraded by acid hydrolysis at pH 3.8, 120 ℃ for 3 h, and at pH 4, 120 ℃ for 4 h, respectively. The resulted acidic hydrolysates were separated by gel permeation chromatography to get oligosaccharide fractions, and the oligosaccharides were obtaind by repeated purification on gel chromatography. The structures of the oligosaccharides were characterized by analyses of infra-red spectroscopy (IR), nuclear magnetic resonance spectrometry (NMR), and mass spectrometry (MS). Results It was showed that the derived oligosaccharides were in saturated structures with the same structure characteristics as the original alginate macromolecule. Conclusion The method des cribed in this paper is a effective and convenient approach for the preparation of low-molecular-weight oligosaccharides from alginate.
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Objective: To investigate the antitumor activity and immune enhancing effect of glycoprotein from Chlamys (Azumapecten) farreri (FGP). Methods: S 180 bearing mice were used as animal model. The effects of FGP on tumor weight and immune function were observed in S 180 bearing mice. Results: The results showed that FGP could markedly decrease tumor weight and promote the phagocytic rate of macrophages, NK activity and increase weight of immune organs. Conclusion: FGP can inhibit the growth of S 180 tumor and enhance cell mediated immune function. The antitumor effect of FGP may be related to its immune enhancing activity.