OBJECTIVE:To study the effects of propofol on invasion of hepatoma carcinoma HepG2 cell. METHODS:MTT method was used to detect the viability of HepG2 cells which were cultured with 0(negative control),1,3 and 10 μg/ml propofol for 48 h. The ability of cell invasion was detect by Tranaswell method. The phosphorylation level of nuclear factor-κB p65(NF-κB p65) and the expression of matrix metalloproteinase 2 (MMP-2),MMP-9,E-cadhherin and Snail were detected by Western blot method. RESULTS:Compared with negative control,1,3,10 μg/ml propofol inhibited the cell viability and invasion,down-regu-lated the expression of Snail and the phosphorylation level of NF-κB p65,and up-regulated the expression of E-cadherin (P<0.01). 3,10μg/ml propofol down-regulated the expression of MMP-2 and MMP-9(P<0.01). Above effects depended on drug con-centration(P<0.01). CONCLUSIONS:Propofol can suppress HepG2 invasion,which might be related to the inhibition of NF-κB/Snail signal pathway.

Objective To establish the method to determine the content of ?-sitosterol in Rhizoma Heterosmilacis Japonicae.Method ?-sitosterol in Rhizoma Heterosmilacis Japonicae was determined by HPLC.Chromatographic column was sinachrom ODS-BP 5 ?m column(4.6 mm?250 mm).Mobile phase was methanol-0.1% H3PO4 liquor.Detection wavelength was 210 nm.Flow rate was 1.0 mL/min.Result ?-sitosterol showed a good linear curve in the range of 0.22～8.8 ?g(r=1.000 0).The average recovery was 96.5% with RSD of 1.15%.Conclusion The method was accurate and reproducible,and can be used to control the quality of Rhizoma Heterosmilacis Japonicae.

Objective To observe the antioxidant activities of whey protein peptides (WPP) in aged mice.Method The subacute aged model mice were made by neck back subcutaneous injection of D-galactose every day.Compared with VE as positive control,the mice were given three different doses of WPP,100,200,400 mg/(kg bw?d) respectively,the effect of WPP on the content of catalase (CAT),malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxidase (GSH-PX)in serum,liver and brain were observed after 45 d.Results The CAT,SOD and GSH-PX activities in aged model group were significantly decreased,but MDA was significantly increased as compared to normal mice.While in the aged mice treated with WPP 200 and 400 mg/(kg bw?d),the activity of CAT,SOD and GSH-PX were significantly increased and the content of MDA significantly decreased as compared to aged mice.Conclusion WPP shows dose-dependant antioxidant effect in aged mice.

Objective To investigate the effect of STAT3 knockdown on the sensitivity of breast cancer cells with drug-resistant to adriamycin (MCF-7/ADR). Methods Levels of STAT3 and p-STAT3 in MCF-7/ADR and MCF-7 cells were detected by Western Blot. The MCF-7/ADR cells were infected with lentivirus expressing STAT3-shRNA and the negative control vectors in the STAT3-RNAi group and NC group, respectively, wihle the cells in the blank group received no treatment. The transfection efficiency was observed with fluorescence microscope, the mRNA level of STAT3, protein levels of STAT3 and p-STAT3 were detected by qRT-PCR and Western Blot, respectively. MCF-7/ADR cells were treated with different concentrations of adriamycin for 48 hours, cell proliferation was detected by MTT assay and cell apoptosis was detected by flow cytometry. Results Levels of STAT3 and p-STAT3 in MCF-7/ADR cells were significantly higher than those in the MCF-7 cells (P < 0.05). The levels of STAT3 mRNA, STAT3 and p-STAT3 in the STAT3-RNAi group were significantly lower than those in the Con group and the NC group (P<0.05, respectively). The Adriamycin IC50 in the Con group, NC group and STAT3-RNAi group was (56.1 ± 3.00)ug/mL,(54.9 ± 11.9)ug/mL and (7.6 ± 0.2)ug/mL, respectively. The flow cytometry results showed that the cell apoptosis in the Con group, the NC group and the STAT3-RNAi group was (10.5+0.7)%, (11.7+0.7)%and (34+3.1)%, respectively. Conclusion LV-shRNA-STAT3 can significantly inhibit STAT3 expression and enhance the sensitivity of breast cancer cells to adriamycin, and the underlying mechanism may be related to cell apoptosis.

Purpose To investigate the gene expression and significance ofβ-catenin, Cyclin D1 and Notch1 in primary breast cancer stem cells ( BCSC) and matched lymph node metastasis stem cells. Methods 30 cases of breast invasive ductal carcinoma and matched metastasis lymph nodes were made into single cell suspensions, then BCSC were separated from them by immunomagnetic sor-ting. β-catenin, Cyclin D1 and Notch1 gene expressions of Wnt, Notch signaling pathway were detected by real time PCR. ResultsThe expression of β-catenin in primary BCSC and matched lymph nodes metastasis stem cells had statistically no differences ( P >0.05), while the expression of Cyclin D1 and Notch1 in matched lymph nodes metastasis stem cells were significantly higher than the expression in primary BCSC (P<0.01, respectively). Conclusion Compared with the primary cancer stem cells, Cyclin D1 and Notch1 activation in metastasis cancer stem cells are in higher level, which leades to a higher capability of invasion and metastasis, which may be a new therapeutic target.

Objective To investigate the effects of Shuangligan and Glycine on Thl/Th2 balancing on severe acute pancreatitis ( SAP) in rats. Methods Thirty-two Wistar rats weighing (260 ± 20) g were randomly divided into sham operation (SO) group, SAP group, SAP + Slg (with the treatment by Shuangligan) group and SAP + Gly (with the treatment by Glycine ) group. Each group included 8 rats, which accepted different treatment according to the experimental design. Changes of plasma level of endotoxin ( ET) and serum amylase (AMY) and the effects of Shuangligan and Glycine on Thl/Th2 ratio at the 24th hour after operation were observed respectively. Results The plasma endotoxin (ET) level ( (0. 67 ±0. 11) EU/ml),proinflammatory cytokine (INF-γ:(8.43 ± 0.86) ng/L, IL-12: (8.26 ± 1.97) ng/L) and Thl/Th2 ratio (0.36 ± 0.07) in SAP group were significantly higher than those in SO group( ET: (0. 44 ±0.07) EU/ml, INF-γ: (3. 80 ±0. 55) ng/L, IL-12: (3. 34 ± 1. 34)ng/L,Thl/Th2 ratio (0. 24 ±0. 05) ) (P <0. 05). Compared with SAP group, SAP + Slg and SAP + Gly group had remarkably decreased plasma ET level ( (0. 57 ± 0. 08,0. 52 ± 0. 04) EU/ml) (P < 0. 05) and the Thl/Th2 ratio reached equilibrium ( SAP + Slg group; (0. 29 ± 0. 04 ), SAP + Gly group: (0. 25 ± 0. 06 )) . Conclusions In the earlier stage of SAP, the rising plasma ET level may cause the overreaction of the cell mediated immune response, which leads to the aggravated damages in tissue cells. Our data indicates that Shuangligan and Glycine can restrain the formation of intestinal endotoxemia and alleviate or prevent the tissue injuries.

Objective To study the antidepression action of curcumin and to explore its mechanism. Methods Mouse reserpine-induced depression model and mouse tetrabenazine-induced acquired despair model were used to observe the effect of curcumin on relieving depression. According to the hypothesis of monoamine, the effect of curcumin on activating monoamine, and inhibiting reuptake of monoamine neurotransmitters and monoamine oxidase inhibitor (MAOI) were observed. Results Curcumin in the dose of 50 mg/kg and more can relieve melancholic symptoms in mice induced by reserpine and tetrabenazine. Curcumin had no direct activation on monoamine either had no obvious inhibition on reuptake of monoamine neurotransmitters of noradrenalin, 5-hydroxytriptamine and dopamine . However, Curcumin had an obvious effect on improving rat forelimb spasm induced by tryptamine hydrochloride. Conclusion Curcumin acts like a kind of monoamine oxidase inhibitor, which exerts antidepression action by inhibiting monoamine oxidase activity and increasing the concentration of monoamine neurotransmitter in brain.

Objective To observe the histopathologic injury of small intestine and intestinal permeability in chronic renal failure (CRF) rats. Methods Twenty male Sprague-Dawley rats were randomly assigned to CRF group (n=10) and control group (n=10). 5/6 nephrectomy was used to establish CRF rats, while sham operation for control. Blood biochemistry was regularly monitored until CRF model was successfully established. The model rats were fed with lactulose (L) and mannitol (M) through intragastric administration. Urine was collected after 6 hours, and the concentration of lactulose and mannitol in urine was measured using high pressure liquid chromatograph with refractive index detector (HPLC-RID), and the ratio of urinary excretion of L/M was calculated to evaluate intestinal permeability. Small intestinal mucosa were stained by hematoxylin-eosin (HE) and observed with light microscope (villus height, thickness of muscle layer and villus count), histological damage score was used to evaluate intestinal injury. Results The L/M ratio of CRF group was higher than that of control group (1.75±0.11 vs 1.20±0.06, P<0.01). The small intestinal mucosal villus height and thickness of muscle layer in CRF group were higher (P<0.01), and the number of villi was lower compared to control group (P<0.01). The score of histopathologic intestine damage of CRF group was higher than that of control group (1.00±0.71 vs 0, P<0.01). Conclusion The intestinal permeability of CRF rats is increased with varying degrees of intestinal damage.

Objective: To study the pharmacokinetics and brain/plasma concentration ratio of nortriptyline at multiple doses in mice which were pre-treated with physiological saline, piperine and verapamil. Methods: A total of 216 male Kun Ming mice[(25±3) g] were equally divided into 4 groups randomly. Each group was intragastrically administered physiological saline (B), piperine (170 μg/kg), piperine (5 mg/kg) and verapamil (5 mg/kg) for 8 days. On the 8th day, 1 h atfer giving the above drugs, each mice was intraperitoneally injected nortriptyline (13 mg/kg). The mice were sacriifced by picking off eyeballs at the time intervals of 5, 15, 30 min, and 1, 2, 4, 6, 8 and 12 h, andthe cerebra were collected and weighted. Nortriptyline in mouse plasma and brain was determined by HPLC-MS/MS. The pharmacokinetic properties of the plasma, brain and brain/plasma were calculated. Results: hTe AUC0-12 h of brain/plasma concentration ratio in the 170 μg/kg piperine group was significantly lower than that in the other groups (P<0.05), while the AUC0-12 h of brain/plasma concentration ratios in the 5 mg/kg piperine group and the verapamil group were not signiifcantly different from those of untreated mice. Conclusion: Piperine (170 μg/kg) may induce P-glycoprotein expression in the blood-brain barrier, while piperines at 5 mg/kg has no influence on P-glycoprotein expression in the blood-brain barrier.

AIM: To observe the changes of cardiomyocytes after stimulation by TNF-?, IL-1?, LPS.METHODS: Cardiac ventricular myocytes were cultured in vitro. Different doses of TNF-?, IL-1?, LPS were added to stimulate the cardiomyocytes, the hypertrophy of cardiomyocytes 8 h, 24 h, and 48 h after stimulation was determined and the apoptosis were also observed 24 h, 48 h, 72 h after stimulation. RESULTS: Compared to the normal myocytes, the cardiomyocytes were hypertrophied after stimulation by 10 ?g/L, 15 ?g/L of TNF-?, 20 ?g/L, 100 ?g/L of IL-1? and 10 mg/L, 15 mg/L, 20 mg/L of LPS, and the effect was dose-dependent, the strongest effect was showed in 24 h. Moreover, 20 ?g/L of TNF-?, 100 ?g/L of IL-1? and 30 mg/L of LPS caused cardiomyocyte apoptosis, especially in 72h. CONCLUSION: TNF-?, IL-1?, LPS induced the cardiomyocyte hypertrophy and apoptosis, suggesting the inflammation may be the main cause of cardiovascular disease.