Objective To analyze the enzyme kinetics of active ingredient of Buddleja lindleyana(AIBL)against Oncomela?nia hupensis,the intermediate host of Schistosoma japonicum. Methods O. hupensis snails were placed in 1 000 ml of 3.55 mg/L AIBL solution for 24,48 h and 72 h,respectively,and the enzyme kinetics of alanine aminotransferase(GPT)was deter?mined by Reitman?Frankel assay,lactate dehydrogenase(LDH)by the chemical inhibition lactic acid substrate method,alka?line phosphatase(AKP)by the disodium phenyl phosphate colorimetric method,acetylcholine esterase(AChE)and malate de?hydrogenas(MDH)by ELISA,and succinate dehydrogenase(SDH)by the phenazine methyl sulfate reaction method(PMS)in the soft tissues of O. hupensis before and after AIBL treatment. Results Following exposure to 3.55 mg/L AIBL solution for 24 h, the GPT,LDH,and AKP activities significantly improved in the soft tissues of O. hupensis,while the SDH and MDH activities were significantly lowered in the head?foot and liver. However,AIBL treatment did not cause significant effect on AChE activity in O. hupensis. Conclusions AIBL causes significant damages to O. hupensis liver and can efficiently act on anaerobic and aer?obic respiration loci,which will hinder energy metabolism,and cause inadequate energy supply in cells used for normal secre?tion,eventually leading to O. hupensis death.

Objective To optimize the fermentation conditions of molluscicidal endophyte LL3026 from Buddleia lindleyana. Method The medium composition and cultivation conditions were optimized by orthogonal and single factor experiments. Re-sults The experiments showed that the conditions of initial pH 3,fermentation temperature 30℃,volume of liquid 100 ml(250 ml Erlenmeyer flask),and 3D-xylitol 0.5 g/L were optimum,and the molluscicidal activity of the fermentation filtrate reached 95%. After three hatches of cultivation,the predicted values were verified by validation experiments. Conclusion Endophyte LL3026 from Buddleia lindleyana has a good molluscicidal activity after the optimization.

AIM:Xilei Powder,a traditional Chinese prescription,has been used to treat wounds for hundreds of years,but the mechanism has not been fully understood.METHODS:The effects of Xilei Powder on fibroblast proliferation,collagen accumulation,matrix metalloproteinases-2,9 ( MMP-2,9 ) activities and tissue inhibitor of metalloproteinase 1 (TIMP-1) production were investigated by MTT,chloramine T method,gelatin zymography and enzyme-linked immunosorbent assays ( ELISA),respectively.RESULTS: The aqueous extract of Xilei Powder significantly promoted fibroblasts proliferation in a time and concentration manner,the population doubling time (125 μg/mL) was 33.8 h,it also significantly (P ＜0.05 ) promoted collagen production.Both of the aqueous and alcoholic extracts could significantly ( P ＜ 0.05 ) increase MMPo2 activity,and also very significantly ( P ＜ 0.01 )promote TIMP-1 production.CONCLUSION: Xilei Powder could promote fibroblasts proliferation,collagen and TIMP-1 production,this might be parts of mechanism to promote wound healing.

AIM:Xilei Powder,a traditional Chinese prescription,has been used to treat wounds for hundreds of years,but the mechanism has not been fully understood.METHODS:The effects of Xilei Powder on fibroblast proliferation,collagen accumulation,matrix metalloproteinases-2,9(MMP-2,9)activities and tissue inhibitor of metalloproteinase 1(TIMP-1)production were investigated by MTT,chloramine T method,gelatin zymography and enzyme-linked immunosorbent assays(ELISA),respectively.RESULTS:The aqueous extract of Xilei Powder significantly promoted fibroblasts proliferation in a time and concentration manner,the population doubling time(125 ?g/mL)was 33.8 h,it also significantly(P

To explore the influence of calculus bovis on the function of primary cultured mice oral fibroblasts, we determined the effects of calculus bovis on the fibroblast proliferation, collagen production, matrix metalloproteinases-2, -9 activities and tissue inhibitor of metalloproteinase-1 production by MTT assay, chloramine T method, gelatin zymography and enzyme-linked immunosorbent assays respectively. The results showed that calculus bovis could significantly inhibit the proliferation of fibroblasts and collagen synthesis in a concentration dependent manner, could significantly (P<0.05) suppress matrix metalloproteinases-2 activity and very significantly (P<0.01) inhibit the production of tissue inhibitor of metalloproteinase-1. In conclusion, the major function of calculus bovis in the process of ulcer healing is not to promote tissue regeneration, the mechanism that calculus bovis inhibits collagen synthesis may be partly due to its ability to very significantly (P<0.01) suppress the production of tissue inhibitor of metalloproteinase-1.
Animals ; Cattle ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cholelithiasis ; chemistry ; veterinary ; Collagen ; drug effects ; metabolism ; Fibroblasts ; cytology ; physiology ; Materia Medica ; pharmacology ; Mice ; Mice, Inbred BALB C ; Mouth Mucosa ; cytology ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; metabolism

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.

Objective To analyze the composition of the aqueous extract of Polygonatum Cyrtonema Hua(PCHE)and evaluate its antitumor activity in vitro and in vivo.Our aim is to provide a theoretical basis for the further development and utilization of Polygonatum Cyrtonema Hua.Methods(1)PCHE was prepared by aqueous extraction,and the chemical composition of PCHE was analyzed by UPLC-Q-TOF/MS and phenol-sulfuric acid method.The inhibitory activity on tumor cells proliferation of PCHE was detected by CCK-8 assay.Cell cycle and apoptosis were detected by flow cytometry,and the expression of apoptosis-related proteins Bcl-2 and Bax was detected by Western Blot.The inhibitory activity of PCHE-containing serum on cell proliferation was detected.(2)A B16 tumor-bearing mice model was constructed and model mice were randomly divided into the model group(saline),the positive drug group(CTX:50 mg·kg-1),and PCHE low-,medium-,and high-dose groups(55.9,111.8,223.6 mg·kg-1),and treated by gavage for 7 days.Changes in body weight and tumor volume of mice were observed during the treatment period.The mice were executed after the treatment,and the histopathological changes of heart,liver,spleen,lung,kidney and tumor were observed by hematoxylin-eosin(HE)staining.The protein expression of Bcl-2 and Bax in tumor tissues was detected by immunohistochemistry(IHC).Results The polysaccharide content of PCHE reached(10.07±1.3)%,and the flavonoid content was(0.044±0.004)%,and thirty-nine components were detected by UPLC-Q-TOF/MS,which contained antitumor components such as flavonoids(baicalein,quercetin,luteolin and rutin),organic acids(ferulic acid)and polyphenols(gallic acid),etc.PCHE exhibited the inhibitory effects on Hela,A549,4T1,B16,MFC and HepG2 cells,among which the inhibitory effect on B16 cells was the most significant(P＜0.001),and PCHE induced cell cycle arrest at G0/G1 phase in B16 cells(P＜0.001).The results of double-staining flow cytometry and Western Blot showed that PCHE significantly promoted apoptosis of B16 cells,decreased the expression of Bcl-2,and promoted the expression of Bax(P＜0.01,P＜0.001).and PCHE constituents absorbed into blood also had an inhibitory effect on B16 cells(P＜0.001).In addition,the results of in vivo activity assay showed that different doses of PCHE could inhibit tumor growth,induce tumor cell necrosis,reduce Bcl-2 expression,and increase Bax expression compared with the model group.Conclusion The ingredients in PCHE are abundant.It contains a variety of antitumor active ingredients,which can inhibit tumor growth,induce tumor cell apoptosis,show strong anti-tumor effects and be worthy of in-depth study.