Objective To study the effects of endotoxin on aldosteron secretion and nuclear factor-κB P65(NF-κB P65)mRNA expression in rat hepatic stellate cell(HSC).Methods Cultured rat HSC were treated with endotoxin of different concentrations. Aldosteron secretion of HSC was detected by radio-immunoassay. NF-κB P65 mRNA expression of HSC was determined by semiquantitative reverse transcription-polymerase chain reaction(RT-PCR).The data were analyzed by variance analysis,t-test and Pearson linear regression analysis. Results Aldosteron secretions of HSC groups treated with 0.01,0.1,1.0 and 10.0 mg/L endotoxin[(4.95±0.35),(5.52±0.32),(6.04±0.60)and(5.16±0.46)μg/L, respectively] were all significantly higher than that in control group(3.655±0.51)μtg/L(t=2.9745,5.8725,6.8465 and 3.2065,respectively;all P＜0.05).NF-κB P65 mRNA expressions of HSC groups treated with 0.01,0.1,1.0 and 10.01 mg/L endotoxin (0.82±0.06、1.07±0.07,1.23±0.06 and 0.96±0.05.respectively)were also significantly higher than that in control group 0.43±0.04(t=5.4776,6.8084,7.9382 and 7.5136,respectively;all P＜0.01).Both aldosteron secretion and NF-κB P65 mRNA expression in HSC groud treated with 10.0 mg/L endotoxin were significantly lower than those in HSC group treated with 1.0 mg/L endotoxin(t=4.3865,3.7246;both P＜0.05).In these treated HSC,aldosteron secretion was positively correlated with NF-κB P65 mRNA expression(r=0.886,P＜0.01).Conclusions Aldosteron secretion and NF-κB P65 mRNA expression in rat HSC could be up-regulated by stimulation with endotoxin, which shows a certain dose-response relationship. This may be one of the important factors of hepatic fibrosis development.

Objective:To re-establish the quality control standard for Weiling granules. Methods:The 4 chief herbs in the prepa-ration, radices paeoniae alba, licorice, rhizoma corydalis and hawthorn were identified by TLC qualitatively. The content of paeoniflor-in in radices paeoniae alba was determined by HPLC. The separation was performed on an Agilent XDB-C18 (250 mm × 4. 6 mm, 5μm)coulme with mobile phase consisting of acetonitrile-0. 1% phosphoric acid solution (10:90). The detection wavelength was 230 nm and the flow rate was 1. 0 ml·min-1 . Results:The spots in TLC were clear without any interference. The linear range for paeoni-florin was 0. 151-1. 212 μg(r=0. 999 9,n=5). The average recovery was 99. 63% and RSD was 2. 01%(n=9). Conclusion:The method is simple and accurate with high reproducibility, which can be used for the quality control of Weiling granules.

Objective To investigate the relationship between dendritic cell (DC)and intestinal endotoxemia in patients with chronic hepatitis B (CHB).Methods Peripheral blood were collected from CHB patients (n = 80)and healthy controls (n = 21 ).Plasma endotoxin (ET)levels,liver function (alanine transaminase,total bilirubin)were detected.According to plasma ET concentration,all CHB patients were divided into two groups:ET positive and ET negative.The peripheral blood mononuclear cells (PBMCs)were isolated and then cultured with recombinant human granulocyte-macrophage colony-stimulating factor ( rhGM-CSF),recombinant human interleukin-4 ( rhIL-4 ),FMS-related tyrosine kinase 3 ligand (Flt3L)and tumor necrosis factor-alpha (TNF-α)to derive DC.The phenotypic patterns were characterized by flow cytometry.The proliferation of T lymphocytes was evaluated with mixed leukocytes reaction (MLR)and the levels of IL-12 and interferon-γ (IFN-γ)produced by DC were analyzed with enzyme-linked immunosorbent assay (ELISA).Comparisons among the two groups and healthy control group were done by single factor analysis of variance.Results Compared to healthy controls,the expressions of CD83,CD80,CD86,human leucocyte antigen (HLA)-DR and the proliferation of allogeneic T lymphocytes by DC were all significantly reduced in CHB patient groups.The expressions of CD83,CD80,CD86,HLA-DR and the activation of proliferation in ET positive subjects were lower than those in ET negative subjects [CD83 (8.25±3.63)% vs(11.39±4.35)% ,CD80 (10.63±4.52)% vs (13.56±5.13)%,CD86 (36.61±16.16)% vs (45.90±15.35)%,HLA-DR (61.65±14.33)% vs (70.35±18.89)%,the activation of proliferation0.812±0.311 vs 1.153±0.324; F=5.123,4.213,3.714,3.323 and 3.125,respectively; all P＜0.05].After cultured for 9 days,the secretions of IL-12 and IFN-γ by DC were significantly lower in CHB patients than in healthy controls [IL-12 (16.99± 6.74)pg/mL vs (44.51±14.56)pg/mL,IFN-γ (10.52±4.19)pg/mL vs (17.94±5.86)pg/mL].The level of IL-12 in the ET positive group was significantly lower than that ET negative group [( 13.14 ±5.71)pg/mL vs (20.98 ± 9.03)pg/mL; F= 3.225,P = 0.016].The level of IFN-γ was not different between two groups [(9.46 ± 3.24)pg/mL vs (11.54 ± 5.20)pg/mL; F = 2.003,P =0.076].Conclusion The intestinal endotoxemia may play a role in DC dysfunction in CHB patients.

We set up the characteristic medical psychology teaching mode based on the talent training objectives for three-year clinical medical majors and the requirements of the course qualification exam.The mode includes the basic psychological and clinical psychological teaching content system based on cases and tasks.Case-based teaching and Problem-based learning were used and research report and net exam were employed as main assessment methods.Online teaching resources were explored through establishing excellent course website,medlical psychology website and psychological test web

Objecfive To investigate the effects of endotoxin on nuclear factor-κB p65(NF-κB p65)mRNA expression and ahtosteron secretion in rat hepatic stellate cells(HSCs).Methods Cultured rat HSCs(HSC-T6)were divided into endotoxin-treated group and control group.Cells in endotoxin-treated group were exposure to 1 mg/ml.endotoxin.Aldosteron secretions of HSCs were determined by radioimmunoassay,and NF-κB p65 mRNA expressions of HSCs were detected by one-step RT-PCR.Results At 6,12,24 and 48 h,aldosteron secretions in endotoxin-treated group were significantly hisher than those in the control group(t=3.063,4.577,6.847 and 9.317,P<0.05),and the expressions of NF-κB p65 mRNA in endotoxin-treated group were also higher than those in control group(t=5.155,6.095,7.875 and 9.313,P<0.01).Aldosteron secretions and NF-κB p65 mRNA expressions in HSCs displayed a positive correlation(r=0.886,P<0.01).Conclusion Endotoxin can up-regulate the aldosteron secretion and NF-κB p65 mRNA expression in rat HSCs,which may be one of the mechanisms of liver fibrosis induced by endotoxin.

Objective: To study the effect of curcumin on cardiac remodeling and heart function change induced by adriamycin in rats. Methods: A total of 50 SD rats were randomly divided into blank control group (n=10), adriamycin group (n=20) and curcumin intervention group (n=20). General conditions of rats, including weight, hair color, diet and activity capacity, were observed. Left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS) and left ventricular end-diastolic dimension (LVEDd) were observed by echocardiography. Rat heart was weighed and levels of cardiac collagen type Ⅰ and Ⅲ were observed using immunohistochemistry method. Results: Compared with blank control group after experiment, there were significant increase in heart weight [(168.53±26)mg vs. (208.34±31)mg], heart weight/body weight [(0.36±0.06) vs. (0.52±0.08)], and significant decrease in body weight [(469.27±34)g vs. (428.47±45)g]; significant increase in LVEDd [(4.87±0.19)mm vs. (7.49±0.83)mm], significant decrease in LVEF [(69.53±6.25)% vs. (41.46±4.38)%] and LVFS [(45.83±3.79)% vs. (17.25±2.92)%] in adriamycin group, P<0.05 all, significant increase in expression rates of collagen type Ⅰ and Ⅲ, P<0.01.Compared with adriamycin group, there were significant increase in body weight (452.46±38)g, significant decrease in heart weight (178.73±38)mg, heart weight/body weight (0.39±0.07); significant decrease in LVEDd (5.96±0.65)mm and significant increase in LVEF (53.12±5.43)% and LVFS (36.57±3.66)%, P <0.05 all; significant decrease in expression rates of collagen type Ⅰ and Ⅲ in curcumin group, P <0.01. Conclusion: Curcumin can improve cardiac remodeling and heart function induced by adriamycin in rats.

Objective:To compare the steam distillation( SD) and supercritical CO2 ( SFE-CO2 ) method in the extraction of vola-tile constituents from such 3 traditional Chinese drugs as cnidium monnieri, radix sileris and atractylodes lancea in compound Kuhuang prescription. Methods:Essential oil was extracted from the 3 traditional Chinese drugs by SD and SFE-CO2 method, respectively. The components were identified by GC-MS, and their relative contents were calculated with peak area normalization method. Results:To-tally 36 components were isolated and identified using SD extraction and 31 components were identified using SFE-CO2 extraction, a-mong them, 22 components were the same and their relative molecular weight mainly concentrated within 200-230. Conclusion: SFE-CO2 method can extract effective components from the 3 traditional Chinese drugs in compound Kuhuang prescription with higher selec-tivity, which is the more suitable extraction method for essential oil from the prescription.

Objective To evaluate the effect of anti-histamine treatment on intestinal endotoxemia and liver inflammation in experimental chronic hepatitis rats.Methods Thirty Wistar rats (15 males and 15 females) were randomly divided into control group (n =8),chronic hepatitis group (n =12) and hepatitis + anti-histamine group (n =10).Chronic hepatitis was induced by subcutaneous injection with 40％ of CCl4,and feeding with low protein,low choline,high cholesterol and high alcohol diet.Antihistamine treatment was given 1 week after the modeling by intragastric administration of ketotifen (1.25 mg/kg).All rats were sacrificed 4 weeks later.Plasma endotoxin,alanine aminotransferase (ALT),total bilirubin (TBil),tryptase,histamine,interferon-γ (IFNγ),iuterleukin (IL)-12,IL-10 and IL-4levels were detected,and the changes in liver histology,the morphology and ultrastructure of mast cells were observed.SPSS 13.0 software package was used for statistical analysis.ANOVA was used for the comparison of measurement data,and SNK method was used for pairwise comparison.Results Plasma endotoxin,ALT,TBil,tryptase,plasma and liver tissue histamine concentrations were (81 ± 19) pg/mL,(186 ± 140) U/L,(10.2±6.2) μmol/L,(0.75 ±0.21) mg/mL,(145 ±52) ng/mL,and (107 ±43) ng/100 mg in chronic hepatitis group,while the above parameters were significantly lower in anti-histamine group except TBil (P ＜ 0.05).Under light microscope,fatty degeneration and fibrosis were formed in liver of chronic hepatitis rats,the hepatic injury was attenuated in anti-histamine group.Toluidine blue stain showed that there was many degranulating and degranulated mast cells filled with purple granula around liver blood vessels and in fiber-interval in chronic hepatitis group,and there were few purple granula in anti-histamine group.The number of mast cells in anti-histamine group was (6.5 ± 1.5)/HP,which was significantly lower than chronic hepatitis group [(10.9 ± 1.6)/HP,P =0.000],but was still higher than that in the control group [(2.2 ± 0.9)/HP,P =0.000].Under electron microscope,the phenomenon of degranulation was severe in chronic hepatitis group and moderate in the anti-histamine group.Compared with the chronic hepatitis group,IL-4 and IL-10 in anti-histamine group were significantly decreased (P ＜0.05),IL-12 was increased (P ＜0.05),but the level of IFN-γ had no significant change (P ＞ 0.05).Conclusion Anti-histamine therapy can significantly improve liver inflammation and alleviate intestinal endotoxemia.

Objective To investigate the correlation between dose and effect of thioacetamide (TAA) on rat model of intestinal endotoxemia. Methods The models of intestinal endotoxemia were induced by three different doses of TAA by gavage administration of TAA 200, 400, 600mg/kg respectively once per day for two days.The doses were given at same time point every day. Each group included 10 rats. The rats in the control group were administrated with 2 mL 0.9% NaCl saline gavage. The death of the rats was observed at 24 hours and 48 hours after administration. The blood samples of the living rats were drawn from abdominal aorta for determining the plasma endotoxin levels, serum alanine aminotransferase(ALT)and aspartated transaminase (AST) levels. The histopathological changes of liver were examined. Single factor analysis of variance was performed and comparision between groups was done using t test. Results No rat in the control group died. Two rats of 200 mg/kg TAA group, five rats of 400 mg/kg TAA group and eight rats of 600 mg/kg TAA group died during the experiment. The mean serum ALT levels of TAA model groups [(305.09±116.78)U/L,(901.67±274.31)U/L,(1454.84±473.49)U/L] were all significantly higher than that of the control group(47.81±22.61)U/L(t=14.583, 25.896 and 20.596, respectively; all P＜0.05). The mean serum AST levels of TAA model groups [(465.88±139.96)U/L, (884.37±250.90)U/L,(1889.23±159.67)U/L] were all significantly higher than that of the control group (69.33±22.04)U/L(t=12.988,18. 455 and 13.542, respectively; all P＜0.05). The mean plasma endotoxin levels of TAA model groups [(0.436±0.110)EU/mL, (0.550±0.095) EU/mL, (0.620±0.057)EU/mL] were all significantly higher than that of the control group (0.103±0.056)EU/mL(t=7.335, 5.260 and 8.191, respectively; all P＜0.05). The histological results of TAA model groups showed hepatic cell degeneration and necrosis in different degrees. Conclusions TAA with 200-600mg/kg is proper to establish the rat model of intestinal endotoxemia. The death rate of rats in the 200mg/kg TAA group is lower than those of other model groups, which suggests that 200mg/kg TAA may be the best dosage for establishing rat model for further studies.

OBJECTIVE:To optimize the water-extraction technology of Compound kuhuang formulation. METHODS:The wa-ter-extraction technology of Compound kuhuang formulation was optimized by orthogonal design,with the total contents of matrine and oxymatrine,the total content of prim-O-glucosylcimifugin and 5-O-methylvisamminol,the content of berberine hydrochloride and extract yield,and with the extraction time,amount of water(3 reflux extractions)and soaking time as the factors;and verifi-cation tests were conducted. RESULTS:The extraction time had significant effect on comprehensive evaluation value (P<0.05). The optimal water-extraction technology was as follows as extraction time of 120 min,water amount of 8,6,and 6 times as the amount of herbs,and soaking time of 20 min. In verification tests,the average total content of matrine and oxymatrine was 3.152 6 mg/g(RSD=1.03%,n=3),that of prim-O-glucosylcimifugin and 5-O-methylvisamminol was 4.977 2 mg/g(RSD=2.27%,n=3),the average content of berberine hydrochloride was 3.345 0 mg/g (RSD=1.19%,n=3) and the average extract yield was 49.23%(RSD=2.43%,n=3). CONCLUSIONS:The optimal water-extraction technology of Compound kuhuang formulation is stable and feasible.