OBJECTIVE：To establish a metho d for sim ultaneous determination of 8 flavonoid glycosides in Sedum bulbiferum . METHODS：HPLC method was adopted to determine the contents of kaempferol- 3-O-β-D-glucopyranoside-（1→2）-α-L-glucopy- ranoside-7-O-α-L-glucopyranoside（KGGR），kaempferol-3-O-β-D-glucopyranosyl-7-O-α-L-rhamnopyranoside（KGR），quercetin-3- O-α-L-rhamnose-7-O-α-L-rhamnoside（QRR），BulbiferumosideⅡ，kaempferol-3-O-（6-coumarinyl）-β-D-glucose-（1→2）-β-D-glu- cose-7-O-α-L-rhamnoside（KcGGR），kaempferol-3-O-（2-β-D-glucose）-α-L-rhamnose-7-O-α-L-rhamnoside（KGRR），kaempferol-3- O-α-L-rhamnoside-7-O-α-L-rhamnoside（KRR），kaempferol-3-O-（6″-acetyl-β-D-glucose）-7-O-α-L-rhamnoside（KaGR）in S. bulbi- ferum. The determination was performed on Waters CORTECS C 18 column with mobile consisted of acetonitrile - 0.1％ phosphoric acid water solution （gradient elution ）at the flow rate of 0.8 mL/min. The detection wavelength was set at 254 nm，and column temperature was 35 ℃. The sample size was 5 μL. RESULTS：The linear range of 8 constituents were 0.013-0.052，0.005-0.018， 0.008-0.031，0.010-0.042，0.009-0.038，0.008-0.030，0.009-0.037，0.032-0.130 μg，respectively（all r were not less than 0.999 0）. The limits of detection were 0.08，0.14，0.11，0.21，0.42，0.35，0.23，0.28 μg/mL，respectively. The limits of quantification were 0.25，0.47，0.38，0.69，1.40，1.17，0.77，0.93 μg/mL，respectively. RSDs of precision ，reproducibility and stability tests （24 h） were all lower than 3％（n＝6 or n＝7）. The average re coveries were 99.67％-104.20％（RSDs＝0.17％-1.59％，n＝6）. Average contents of above 8 constituents in 13 batches of samples were 0.893 8，0.312 6，0.490 8，0.964 9，0.751 2，0.502 2，0.606 2， 1.915 7 mg/g（n＝3）. CONCLUSIONS ： The method is simple， acourate and reproducible ， and can be used for simultaneous determination of 8 flavonoid glycosides in 才〔2016〕5677） S. bulbiferum .

Aim To investigate the effects of starfish saponins(Sfs) on insulin signaling pathway in orotic acid-induced NAFLD rats.Methods 1% orotic acid was used to establish NAFLD model in male Wistar rats for six weeks.The NAFLD rats were randomly divided into two groups(eight rats in each group) and then fed with the corresponding diets: Model group(1% orotic acid)and Sfs group(1% orotic acid containing 0.04% starfish saponins).After starfish saponins feeding for 8 weeks, hepatic lipids content, liver function indices and relevant protein expression in muscle insulin signaling pathway were measured.Results Compared with model group, starfish saponins reduced hepatic lipids content and improved liver functions.In addition, it effectively ameliorated insulin resistance by improving insulin signaling pathway and improved glucose uptake in muscle.Conclusion The amelioration effect of starfish saponins on impaired insulin signaling pathway in muscle is observed in orotic acid-induced NAFLD rats.

Aim To investigate the immunomodulatory effects of sea cucumber fucoidan ( SC-FUC) on macro-phage and the signaling pathways. Methods Cell via-bilities in response to different concentrations of SC-FUC were analyzed by MTT, phagocytosis ability was detected by neutral red,and nitric oxide ( NO) produc-tion was examined by Griess reaction kit. The mRNA expression levels of IL-6 , IL-10 , Toll-like receptors (TLRs) and related signal molecules MyD88, TRIF, NF-κB were assayed by real-time PCR. All the experi-ments were based on murine RAW264. 7 cell line. Re-sults SC-FUC could promote RAW264 . 7 cell prolif-eration, phagocytosis as evidenced by uptake of neutral red and release of NO. The effects were significant at the early stage (6 h and 12 h) . SC-FUC could up-reg-ulate the expression of IL-6 , IL-10 , TLR4 , TLR5 , TLR9. Moreover, mRNA expressions of TLRs signaling molecules were increased, as well as MyD88, TRIF, NF-κB. Conclusions SC-FUC could activate macro-phage, and then promote the immune function by pro-moting production or expression of NO, IL-6, IL-10. It is speculated to be relevant to activated cell surface re-ceptors in macrophage, including TLR4, TLR5, TLR9, and NF-κB signaling pathways.

Aim To investigate the effects of saponin of sea cucumber ( SSC ) on the blood pressure in obese mice. Methods C57BL/KsJ(db/db) mice were ran-domized into 3 groups ( 8 mice each ): model group, low-dose SSC group and high-dose SSC group. Normal C57BL/KsJ mice were used as control. The low and high SSC groups were fed on basal diets incorporated with 0. 02% and 0. 04% SSC. Different treatments were administered for 6 weeks and arterial pressure was measured in the third and sixth weeks. The abundance of renal ACE, ACE2 and REN mRNA was detected by real time PCR . Results Compared with control group, the blood pressure of model group mice was ob-viously raised ( P<0. 01 ) . Low-dose SSC group mice showed lower blood pressure than model group without statistically significant differences, and the blood pres-sure of high-dose SSC group mice was similar to that of control group and significantly lower than model group. ( P<0. 05 ) There were no remarkable differences a-bout ACE and REN mRNA among the groups, howev-er, ACE2 mRNA level was significantly increased in high-dose SSC group. Conclusion SSC plays a vital role in decreasing blood pressure, which probably re-lates to the regulating function of renin-angiotensin sys-tem( RAS) .

Objective To study the sleep improvement function of DHA-PC.Methods The mice were randomly divid-ed into control, vehicle, DHA+Lecithin (60+200 mg/kg) and DHA-PC(50,100,200 mg/kg) groups.Ten mice were enrolled in each group .The mice of control were administered with normal food , the vehicle group was orally given normal saline at the dosage of 0.2 ml/10 g, while both DHA-PC and DHA+Lecithin were orally given corresponding drugs at the dosage of 0.2 ml/10 g.All the groups were treated for 30 days except control group .The direct sleep-inducing test, the test of lengthening sleep time induced by pentobarbital sodium , the test of pentobarbital sodium subthreshold-hypnosis and the test of barbital sodium sleep latency were conducted to observe the inductive effect of DHA -PC.Results Neither the effect on mice body mass nor directly-induced sleep was observed .DHA-PC (50,100, and 200 mg/kg) could prolong sleep time to (56.2 ±13.7),(57.9 ±25.4) and(64.1 ±18.4) min, respectively,compared to vehicle(32.9 ±10.8)min (P<0.05).DHA+Lecithin could not prolong sleep time (38.6 ±11.7)min compared to (32.9 ±10.8)min of vehicle.There was significant difference compared with DHA-PC at the dosage of 200 mg/kg (64.1 ±18.4)min (P<0.05).DHA-PC (200 mg/kg) enhanced pentobarbital sodium subthreshold-hypnosis (70%) compared to vehicle (10%) (P<0.05),so did DHA+Lecithin (60%) compared to vehicle (10%) (P<0.05).Both DHA-PC (200 mg/kg)[(22.9 ±4.1)min ] and DHA+Lecithin [(19.5 ±2.7) min ]could shorten sleep latency compared to vehicle (31.3 ±6.9) min(P<0.01), and the sleep latency of DHA +Lecithin (19.5 ±2.7) min was shorter than that of DHA-PC(50,100 mg/kg).Conclusion DHA-PC has some effect some sleep improvement in mice .

AIM To evaluate the prevention and treatment of N-(2-mercaptopropionyl)-glycine sodium (MPG-Na) and tiopronin (MPG) on acute liver injury. METHODS The experimental mouse model of hepatotoxicity induced by D-galactosamine (Gal) was applied to investigate preventive and remedial effects. In the preventive experiment, the mice were ip administered with MPG-Na or MPG 37.5，75 and 150 mg·kg~(-1), respectively, for 7 d. Gal 800 mg·kg~(-1) was ip given into the mice 30 min after the last administration. In the remedial experiment, the mice were ip given Gal 800 mg·kg~(-1) and 30 min later followed by MPG-Na or MPG 37.5, 75 and 150 mg·kg~(-1) , respectively, for 2 d. The mice were euthanized and serum was prepared 24 h (pre-treatment) or 48 h (post-treatment) after Gal injection. The activities of serum glutamyl pyruvic transaminase (GPT) and glutamyl oxaloacetic transaminase (GOT), the contents of total protein (TP) and albumin (Alb), and the Alb/globulin (A/G) ratio were determined. The liver tissues were collected for histopathological assessment (HE staining) under light microscope. RESULTS Compared with normal control group, the activities of serum GPT and GOT in model group were significantly increased. The injuries such as fatty degeneration and liver cell necrosis were observed. Compared with model group, the activities of GPT and GOT in pre-treatment groups were obviously decreased in MPG-Na 150 mg·kg~(-1) group. In post-treatment groups, the activity of GPT decreased in 3 MPG-Na groups. The contents of TP, Alb and A/G ratio had little change. In addition, MPG-Na alleviated the injuries such as fatty degeneration and liver cell necrosis obviously. Compared with MPG, MPG-Na showed similar effect. CONCLUSION MPG-Na has an obvious protective effect against Gal-induced acute liver injury in mice and the efficiency is equivalent as MPG.

Objective:To investigate the effect of dietary cholesterol on hepatic TG accumulation in rats. Method:Fourteen male Wistar rats were randomly divided into 2 groups and fed 1% cholesterol or cholesterol free AIN76 diets. After 4 w,serum triglyceride(TG) ,total cholesterol(TC) ,high density lipoprotein cholesterol(HDL-C) ,phospholipids(PL) ,glucose and free fatty acid(NEFA) levels were determined. Hepatic lipid concentrations(TG,TC,PL) and the activities and/or mRNA expression of malic enzyme(ME) ,glucose-6-phosphate dehydrogenase(G6PDH) ,fatty acid synthase(FAS) ,phosphatidate phophatase(PAP) ,carnitine palmitoyl transferase(CPT1,2) ,HMG-CoA reductase,acylCoA-cholesterol acyltransferase(ACAT) ,cholesterol 7?-hydroxylase(CYP7A) were also determined. Results:The serum TC and non-HDL-C levels were significantly increased but TG and HDL-C levels were significantly decreased by cholesterol feeding. The concentrations of hepatic TC and TG were 4-20 folds higher in cholesterol group than those in cholesterol free group. The activities of hepatic ME,G6PDH,FAS,PAP and CPT were depressed by cholesterol(40%,70%,50%,15% and 25% respectively) . The mRNA expression of FAS,CPT1,CPT2,and HMG-CoA reductase were down-regulated(35%,30%,50% and 25% respectively) and CYP7? and ACAT were up regulated(6.5 and 1.6 fold) by cholesterol in liver. Conclusion:The dietary cholesterol increases TG accumulation in liver,but dose not stimulates the activity and the gene expressionof hepatic TG synthesis related enzymes.

10000 u,SP2:6 000 u0.05),the melanogenesis and tyrosinase activity were inhibited remarkably(P

Objective To investigate the effects of sepia on stem cells,granulocyte and monocyte progentior cells and peripheral WBC in mice.Methods Different dosages of sepia were given to normal and model mice with hematopoietic system impairment respectively.The numbers of CFU-S,CFU-GM and peripheral WBC in normal and model mice were measured respectively with the method of hematopoietic progenitor cells cultured in vitro and the technique of experimental hematology.Results Sepia could enhance the number of CFU-S,CFU-GM and peripheral WBC in normal mice significantly,resist the decrease of CFU-S,CFU-GM,and peripheral WBC in model mice of hemapoiecsis impaired effectively and promote the restoration of those indices mentioned above in model mice significantly.Conclusion Sepia has significant effects on stimulating granulopoiesis in bone marrow in mice.The mechanism may be related to regulating immunological function and inducement GM-CSF and other sorts of cellular factors,which in turn promote the multiplication differentiation of CFU-S and CFU-GM.