BACKGROUND:The hepatic stellate cells(HSCs)plays a key role in the development of liver fibrosis.Studies have shown that bone marrow-derived mesenchymal stem cell(BMSCs)transplantation can be used to treat liver fibrosis,but the mechanism for reversal of liver fibrosis remains unknown.OBJECTIVE:To explore the mechanism of bone marrow mesenchymal stem cells to regulate the proliferation of HSCs under co-culture in vitro.METHODS:Rat BMSCs and HSCs in the experimental group were cultured in the plastic culture plate(6 holes)to establish the upper and lower double-cell co-culture system.Rat normal fibroblast cell lines were seeded as control group;HSCs were cultured alone as blank group.Cell proliferation was determined by WST8 and cell cycle was determined by flow cytometry.The Cyclin D1 and P27 mRNA expression in HSC was determined by reverse transcription-polymerase chain reaction(RT-PCR)and the level of Cyclin D1 and P27 protein by Western blot.RESULTS AND CONCLUSION:HSCs co-cultured with BMSCs significantly inhibited HSC proliferation compared with the blank and control groups at 24,48,and 72 hours(P ＜ 0.01);Flow cytometry showed that the percentage of G_0/G_1 phase cells of co-culture group was increased but the S phase cells reduced(P ＜ 0.01)compared with the other groups at 72 hours,and BMSCs blocked HSC to convert from G_0/G_1 period to S phase.After HSCs co-cultured with BMSCs for 24 hours,the expression of Cyclin D1 mRNA and protein was reduced,and significantly less than the blank and control groups at 72 hours(P ＜ 0.01);no differences were detected in P27 mRNA expression in each group during the co-culture(P ＞ 0.05).After co-culture of 24 hours,the p27 protein expression was significantly increased compared with the blank and control groups(P ＜ 0.01).BMSCs inhibited the proliferation of HSCs,possibly through inhibiting CydinDI expression,increasing the p27 protein expression to cause cell cyde arresting in G_0/G_1 phase.

Objective: To establish an optimal extraction process of the total glycoalkaloids in Solanum tuberosum L. and then study the anti-inflammatory activity. Methods:The extraction process of the total glycoalkaloids was optimized by orthogonal design. Compared with that of the total glycoalkaloids in Rhizoma Dioscoreae Melanophymatis and fresh potato pieces, the anti-inflammatory ac-tivity of the total glycoalkaloids in Solanum tuberosum L. was evaluated by mouse ear swelling model induced by xylene, rat paw swell-ing model induced by carrageenan, granuloma model caused by cotton and blood capillary permeability experiments. Results:The opti-mal extraction conditions were as follows:the extraction temperature was 65℃, 10-fold amount of methanol was used for twice extrac-tion with 45 min per time. The total glycoalkaloids from the optimal extraction had obvious anti-inflammatory activity,and the effect was related to the content ofα-chaconine. Conclusion:The results show that the order of different factors affecting the extraction rate is ex-traction temperature> extraction time, and the total glycoalkaloids in Solanum tuberosum L. has good anti-inflammatory effects in mice and rats.

Designed experiment in medical functional experiment has become an important way to promote creative thinking and innovation of medical students.We applied different modes of designed experiment in undergraduates of clinical medicine and basic medicine in capital medical university,including classroom designed experiments,proposition designed experiments and free proposition designed experiments.After above reforming implements,creative thinking and innovation ability of medical students were enhanced.It also provided new ideas in future teaching reform in functional experiment.

[Objective] To make a microscopic identification of medicinal material of Herba Rabdosiae Serrae (HRS) from different plant resources. [Methods] The dry leaves of the whole, the middle part and the upper-middle part from the plants of Isodon serra (Maxim.) Kudo, Isodon striatus (Benth.) Kudo, Isodon lophanthoides var. geradianus (Benth.) Hara, and Isodon lophanthoides Hara var. graciliflorus (Benth.) Hara were sliced by different methods. Structure of leaf epidermis, the transverse section of leaf and structure of leaf powder were observed under light microscope. [Results] There existed differences in epidermal cells, stomata distribution, nonglandular hair, glandular hair, small glandular hair and glandular scale of leaf epidermis from four kinds of medicinal plants of Herba Rabdosiae Serrae . And there also existed differences in epidermal cells, anticlinal wall and external-section wall of epidermal cells, nonglandular hair, glandular hair, glandular scale, stomata distribution, mesophyll, main vessels, and vascular bundle of leaf transverse section, as well as in epidermal cells, stomata distribution, glandular scale, nonglandular hair, glandular hair and ducts of leaf powder from four kinds of medicinal plants of Herba Rabdosiae Serrae. [Conclusion] Medicinal plants of different kinds of Herba Rabdosiae Serrae have different microscopic identification structure in leaf epidermis, leaf transverse section and leaf powder.

Objective To compare the influence of different extraction technology on the pharmacodynamic actions of drug pair of Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba.Methods The influences of water extraction,alcohol extraction,and SFE-CO2 extraction on pharmacodynamic actions of drug pair of Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba were evaluated by mice intestine propulsive motility test,rats repelling acute gastric mucosa damage test,mice twisting test and hot-plate test.Results The paired drugs extracted by SFE-CO2 combining with water extraction or alcohol extraction had better pharmacological actions.Conclusion This study provides a scientific basis for optimizing the extraction technology of drug pair of Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba.

Objective To develop a quality standard for Dongfengju oral liq ui d (DOL). Methods Atalantia buxjfolia, Radix Scrophulariae, Fructus Aurantii in DOL was identified by TLC. Results The obtained TLC spots of Atalantia buxj folia, Radix Scrophulariae, Fructus Aurantii in DOL occurred at the correspond ing positions compared to the controls. Conclusion The method is proved to be simple, sensitive, precise and reproducible and can be used for the quality con trol of DOL.

[Objective] Antifungal action of fat-soluble extracts from Bulbus Allii Cepae (BAC) by different extraction technology on 16 strains of dermatophytes was observed to supply evidence for the optimization of extraction technology for BAC. [Methods] Four methods were used for the extraction of fat-soluble extract from BAC and the antifungal action of the extract was tested by medium dilution method. [Results] Fat-soluble extract freshly extracted from BAC by ether had a strong in-vitro antifungal action and its minimum inhibitory concentration (MIC) was not over 6.25 mL?L-1 against 10 strains of fungi including Shanghai Epidermophytom floccosum and its MIC against other strains ranged 12.5 -50.0 mL?L-1. The minimum fungicidal concentration of fat-soluble extract freshly extracted from BAC by ether was as the same as MIC. Fat-soluble extract freshly extracted by ether had no antifungal action after water removal by heating. [Conclusion] The extraction technology of freshly extracted by ether without heating is a better method for BAC and the fat-soluble extract extracted by this method has a better antifungal action.

Objective To establish a method for the determination of berberine and salvianolic acid B in Fuyankang En- ema.Methods The chromatograhie separation was performed on a Kromasil C_(18) column (250 mm?4.6 mm,5?m) with a gradient elution of acetonitrile-0.04 mol/L potassium dihydrogen phosphate solution containing 0.1% phosphoric acid.The flow rate was 1.0 mL/min and column temperature was set at 30℃.The detection wavelength was 230 nm.Results There was a good linearity in the range of 0.0924～2.3100?g (r=0.999 9) for berbeine,and 0.1000～2.5000?g (r=0.999 9) for salvianolic acid B.The average recoveries of berberine at high,middle and low concentra- tions were 100.40 %,99.51% and 99.15 % respectively with RSD being 0.59 %,0.67 %and 1.02 % (n=3) re- spectively.The average recoveries for salvianolic acid B were 101.52 %,99.32 % and 98.68 % respectively with RSD being 0.60 %,0.92 % and 1.00 % (n=3),respectively.Conclusions This method is convenient,specific and re- producible for the determination of berbeine and salvianolic acid B in Fuyankang Enema by HPLC.

Objective To establish the HPLC fingerprint for the quality control of Yanhuanglian Injections.Methods HPLC Chromatography method was used.The conditions included a Shim-pack CLC-ODS column(250 mm?6.0 mm,5 ?m),the gredient elution was adopted with acetonitrile-buffer solution(1∶1),the detection wavelength was at 285 nm,and the flow rate was 1.0 mL/min.The Operating Standard of Similarty Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica(Version 2004A)was used to calculate.Results Similarity of 13 batches of injections was over 0.95,the fingerprint of Yanhuanglian Injections was established,and 12 common peaks were indicated.Conclusion This method can be applied to the quality control of Yanhuanglian Injections.

Objective To study the effects of ganoderma spore oil on Friend murine leukemia virus(Fr.MuLV)and its mechanism.Methods The BALB/C female mouse was infected with Fr.MuLV,body weight,spleen index,thymus index,the percentages of CD4+,CD8+,and CD4+/CD8+ were mea-sured.Results Compared to the control group,body weight extenuation,splenomegaly,and atrophy of thymus gland severely in the model group,the percentages of CD4+,CD8+,and CD4+/CD8+ were significantly decreased in the model.Body weight,spleen index,and thymus index(P