Objective To investigate the effect of bifidobacteria on dextran sulphate sodium(DSS)- induced acute ulcerative colitis in mice.Methods Thirty BALB/C mice were randomly divided into nor- mal control group (n=10),0501 strain group (n=10) and c122 strain group (n=10).Fifty BALB/ C mice received 5% dextran sulphate sodium(DSS) for 7 days to induce ulcerative colitis.The mice were then divided to model group,negative control group(perfused with 0.9 NaCl solution ),positive control group(perfused with SASP of 20 mg/ml),DSS + 0501 strain group(perfused with 1?10~9 CFU/ml bifidobacteria 0501 strain solution and DSS + c122 strain group (perfused with 1?10~9 CFU/ml bifidobacteria c122 strain solution).All mice were sacrificed 9 days later.The colon specimens were measure by histoehemical staining with H-E.The expressions of interleukin-10 (IL-10) and its protein were detected by RT-PCR and immunohistochemistry respectively.Results The degree of colon inflam- mation in mice both in DSS+ 0501 strain and DSS+ c122 strain groups were aggravated and expressions of IL-10 mRNA and protein were reduced compared to model group.No colon inflammation was found in 0501 strain and c122 strain groups.Conclusion Some strain of bifidobaeteria may aggravate colon in- flammation in mice when mucosal harrier is destroyed.

Background The select of supporter is critical for the construction of tissue engineering cornea.Many carrier carl be utilized in the construction of tissue engineering cornea,but de-cellular corneal matrix is known to be one of optimal supporters.Objective Present study was to investigate the characteristics of de-cellular corneal matrix of porcine of structure and biocompatibility for rabbit cornea stroma and limbal epithelial ceHs. Methods The porcine cornea was prepared as de-cellular corneal matrix of porcine by the application of detergent enzyme combined process.The corneal epithelial cells,keratocyte and endothelial cells of porcine were removed completely and stored in -20℃ refrigerator after sterilization.The morphology of de-cellular corneal matrix of porcine was examined by hematoxylin-eosin staining under the light microscope.The structure characteristics of de.cellular corneal matrix of porcine under the scan electron microscope,and its physics features were investigated by the evaluation of water content,strength,expansion and transparency.The de-cellular corneal matrix of porcine were implanted to cornea stroma of rabbit and co-cultured with rabbit corneal epithelial cells for 4 weeks in vitro to assess the keracyte compatibility. Results The epithelial cells,keratocyte and endothelial cells of porcine were removed completely by trypsogen digestion.The collagen fibril network and collagen plates paralleled to corneal surface under the light microscope.The water content,strength,expansion。Ratio of light transparency of de-cellular corneal matrix of porcine were similar to normal porcine cornea.After implantation of de.cellular comeal matrix of porcine into rabbits corneal stroma,the edema of tissue was found in one week,and edema disappeared on two weeks and became clear on four weeks after surgery.The de-cellular eorneal matrix attached to rabbit cornea stroma well.No inflammatory eell and new vessel were found after surgery.The co-cultured rabbit corneal epithelial cells differentiated and proliferated on the surfaee of de-cellular corneal matrix and showed positive response for CK3.No statistically significant differences were found in the water content,strength,expansion of de-cellular cornea matrix of porcine among the normal,before dehydration,2 and 4 hours after dehydration cornea matrix(P＞0.05).However,the transparency was much better in the corneal matrix with 2-hour,4-hour dehydration in comparison with non-dehydration one(P＜0.05). Conclusion The structure features of de-cellular cornea matrix of porcine are similar to normal porcine cornea.Good biocompatibility is proved for xenogenesis of rabbit cornea.

Six flavonol glycosides were isolated and calibrated from Ginkgo biloba extract, and then used to calibrate the content in 2 baiches of G. biloba reference extract, so was rutin. RSD values of rutin, kaempferol-3-O-rutinoside, kaempferol-3-O-rhamnoside-2-glu- coside, quercetin-3-O-rhamnop-yranosyl-2-O-(6-O-p-coumaroyl)-glucoside, kaempferol-3-O-rhamnopyranosyl-2-O-(6-O-p-coum-aroyl) - glucoside were around 1.1%-4.6%, nevertheless, RSD values of quercetin-3-O-glucoside and isorhamnetin-3-O-rutinoside were more than 5%. According to the results, the reference extract of G. biloba can be used as the substitute to determine rutin, kaempferol-3-O- rutinoside, kaempferol-3-O-rhamnoside-2-glucoside, quercetin-3-O-rhamnopyranosyl-2-O-(6-O-p-coumaroyl)-glucoside and kaempferol-3-0-rhamnopyranosyl-2-O-(6-O-p-coumaroyl)-glucoside instead of corresponding reference substances. So reference extract in place of single component reference in assay is feasible.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonols ; chemistry ; isolation & purification ; Ginkgo biloba ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

The present study was undertaken to observe the effect of exogenously administered melatonin on the intensity of β-endorphin (β-Ep) immunoreactivity of the neuron in the arcuate nucleus of rat hypothalamus with an aim to explore the possible mechanisms of the analgesic effect of melatonin. The experimental rats were divided into two groups, one injected intraperitoneally with melatonin (110 mg/kg) and the other with only a vehicle. One hour after injection, the brain was processed for coronal sections, which were stained with immunohistochemical ABC technique. The integral optical density (IOD) and mean optical density (OD) of the stained sections were measured with a computer-assisted image-processing and analytical system. β-Ep immunoreactivity was much decreased in the sections treated with melatonin and the IOD and OD were also decreased significantly (P<0.01; P<0.05). The above results suggest that melatonin may result in a decrease of β-Ep content in the arcuate nucleus, as a result of increased β-Ep release induced by administration of melatonin. It is likely that the analgesic effect of melatonin may be in part mediated by the release of β-endorphin from the arcuate nucleus.

Plant active components characterized of many different structures and activities on multiple targets, have made them to be the important sources of inhibitors on HIV-1. For finding leading compounds with new structure against HIV-1, three key HIV-1 replicative enzymes (reverse transcriptase, protease and integrase) were used as screening models. The in vitro activities of 45 plant derived components isolated from Schisandraceae, Rutaceae and Ranunculaceae were reported. Within twelve triterpene components isolated, eight compounds were found to inhibit HIV-1 protease, in these eight active compounds, kadsuranic acid A (7) and nigranoic acid (8), inhibited both HIV-1 protease and integrase; Among fifteen lignans, meso-dihydroguaiaretic acid (15) and kadsurarin (16) were active on HIV-1 reverse transcriptase, and 4, 4-di(4-hydroxy-3-methoxyphenly)-2, 3-dimethylbutanol (13) active on HIV-1 integrase. All of the six alkaloids, seven flavones, and five others compounds were not active or only with low activities against HIV-1 replicative enzymes. Further studies of the triterpene components showing strong inhibitory activities on HIV-1 were warranted.
Alkaloids ; chemistry ; isolation & purification ; pharmacology ; Anti-HIV Agents ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Flavones ; chemistry ; isolation & purification ; pharmacology ; Guaiacol ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; HIV Integrase ; drug effects ; HIV Protease ; drug effects ; HIV Reverse Transcriptase ; antagonists & inhibitors ; Lignans ; chemistry ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Ranunculaceae ; chemistry ; Rutaceae ; chemistry ; Schisandraceae ; chemistry ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

OBJECTIVETo study the culture of adventitious root of Salvia miltiorrhiza in vitro systemically.

METHODEffects of sucrose concentrations, medium pH, inoculum size and plant growth regulators on adventitious root growth and secondary metabolites production in S. miltiorrhiza were investigated.

RESULTWith the increase of initial sucrose concentration, adventitious root growth rates increased and tanshinone II A content decreased, while content of protocatechuic aldehyde showed a broken line change and its highest production was obtained under 30 g x L(-1) sucrose in the medium. As for the effect of medium pH, medium pH of 6.5, 5.5 (or 6.0) and 5.8 was favorable for adventitious root growth, tanshinone II A and protocatechuic aldehyde synthesis respectively. Furthermore, adventitious root growth, rate was greatly increased when inoculum size was 2.5%. MS medium added with 0.5 mg x L(-1) KT was much favorable for tanshinone II A and protocatechuic aldehyde accumulation.

CONCLUSIONParameters including sucrose concentrations, medium pH, inoculum size and plant growth regulators have distinct effects on the in vitro culture of adventitious root growth and secondary metabolites synthesis of S. miltiorrhiza.

Benzaldehydes ; metabolism ; Catechols ; metabolism ; Culture Media ; Diterpenes, Abietane ; Hydrogen-Ion Concentration ; Phenanthrenes ; metabolism ; Plant Growth Regulators ; pharmacology ; Plant Roots ; growth & development ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Salvia miltiorrhiza ; growth & development ; metabolism ; Sucrose ; Tissue Culture Techniques

This study is to investigate the antitumor activity of CIP-36 on multidrug resistant human oral squamous carcinoma cell line (KBV200 cells) in vitro and the possible anticancer mechanisms. MTT assay, Hoechst fluorescein stain, RT-PCR and immunohistochemistry were carried out on KBV200 and KB cells. The growth of many tumor cells was obviously inhibited by CIP-36, especially the multidrug resistant cells KBV200. Obvious apoptosis could be observed in the Hoechst 33342 staining experiments. The results of RT-PCR showed that the levels of p53, p21, caspase-3 and bax mRNA increased, and meanwhile the expression of mdr-1 and bcl-2 mRNA decreased in a dose-dependent manner. The data were significantly different from that of vehicle. The expression of P-gp significantly decreased with the increasing dosage of CIP-36 examined by immunohistochemistry. It can be concluded that CIP-36 could change resistance-related genes and proteins to overcome multidrug resistance in the KBV200 cell line.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Caspase 3 ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; KB Cells ; Mouth Neoplasms ; metabolism ; pathology ; Podophyllotoxin ; administration & dosage ; analogs & derivatives ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins p21(ras) ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

A new UPLC method was developed for the simultaneous determination of eleven characteristic flavonoid glycosides in Ginkgo biloba leaves. The natural occurrence of flavonoid glycosides in Ginkgo biloba leaves within one vegetative season was investigated for the first time. The analysis was performed on an Agilent ZORBAX Eclipse Plus C18 column (50 mm x 4.6 mm, 1.8 microm), the mobile phase A was acetonitrile, the mobile phase B was 0.4% phosphate aqueous solution in a gradient elution at a flow rate of 0.6 mL x min(-1), the detection was carried out at 360 nm. The result showed that eleven flavonoid glycosides had good linearity with good average recovery, separately. The method was proved to be accurate, rapid and good reproducible for the quality evaluation of Ginkgo biloba leaves, and provide an easy and rapid means for the quantitative analysis of flavonoid glycosides and their content fluctuation with seasons.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; chemistry ; Flavonoids ; analysis ; chemistry ; Ginkgo biloba ; chemistry ; Glycosides ; analysis ; chemistry ; Molecular Structure ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Seasons

OBJECTIVETo explore the diagnostic efficiency of circulating antigen using the TM5.28 mAB-biotin-avidin system for the detection of schistosomiasis japonica.

METHODSA mAb-biotin-avidin system was set up using a TM5.28 mAB which was prepared against a gut associated antigen of Schistosoma japonicum. Detection was performed on the sera from 50 acute schistosomiasis patients, 224 chronic patients, 49 advanced patients and 46 schistosomiasis patients who were followed up at 6 months and 12 months post treatment. In addition, 19 cases of clonorchiasis, 31 cases of paragonimiasis, 23 cases of hepatitis B and 100 healthy individuals were also included.

RESULTSThe system showed sensitivity of 83.1% and specificity of 94.0% when applied to detect chronic schistosomiasis and healthy persons respectively, while 94.0% to acute schistosomiasis. The Youden's index of the system was 0.771. The rate of cross-reaction to paragonimiasis, clonorchiasis and hepatitis B was 12.9%, 15.8% and 13.0% respectively. The rates of negative turning were 43.9% and 62.1% respectively in chronic schistosomiasis at the 6 month and 12 month intervals after treatment. Geometric mean of the OD values also decreased from 0.172 before treatment to 0.081 at 6 months and 0.068 at 12 months after treatment with a reduction rate of 60.30%. The detection rate in the heavy infected population reached a maximum of 90.0%. This was similar in moderate and light infected populations, i.e., 83.9% and 82.1%, respectively.

CONCLUSIONThe TM5.28 mAb-biotin-avidin system showed a relatively high efficiency in the diagnosis of schistosomiasis and a high negative turning rate after treatment. It is, therefore, a valuable tool for the estimation of prevalence in endemic populations, as well as individual diagnosis and for assessing the effect of chemotherapy.

Animals ; Antibodies, Helminth ; immunology ; Antibodies, Monoclonal ; immunology ; Avidin ; immunology ; Biotin ; immunology ; Cell Fusion ; Humans ; Mice ; Mice, Inbred BALB C ; Schistosomiasis japonica ; diagnosis ; immunology ; Serologic Tests

OBJECTIVETo study the role of platelet-derived growth factor (PDGF) in the pathogenesis of preeclampsia (PRE).

METHODSThirteen normal and 20 PRE late-pregnancy women were enrolled in this study. The serum PDGF-BB levels were measured with enzyme-linked immunosorbent assay, and the expression of PDGF-B mRNA in the decidual blood vessel was determined using in situ hybridization.

RESULTSPDGF-BB levels in PRE group was significantly higher than that in normal pregnant women (83.54 -/+34.52 vs 39.61-/+18.20, P<0.001), and the expression of PDGF-B mRNA in decidual blood vessel was also significantly higher in PRE group (P<0.001), showing a positive correlation between serum PDGF and PDGF-B mRNA expression (r=0.603, P<0.001).

CONCLUSIONPDGF is associated with the pathology of decidual blood vessel. Elevated serum PDGF levels and PDGF-B mRNA expression in the decidual blood vessel may play an important role in the pathogenesis of PRE.

Adult ; Decidua ; blood supply ; Female ; Gene Expression Regulation ; Humans ; Platelet-Derived Growth Factor ; genetics ; metabolism ; Pre-Eclampsia ; blood ; genetics ; metabolism ; pathology ; Pregnancy ; Proto-Oncogene Proteins c-sis ; RNA, Messenger ; genetics ; metabolism