OBJECTIVE:To establish a RP-HPLC method for determining rhein in plasma.METHODS:Column:Spherisorb C18(4.0mm?250mm,5?m),Mobile phase:0.1% of phosphoric acid(pH 3.0)-methanol(30∶70,V/V),Flow rate:1.0ml/min;Detection wavelength:435nm.RESULTS:Rhein was completely separated and no interference was found.The peak areas of rhein within the range between 0.032～4.8?g/ml showed good linearity of the concentrations,r=0.9 990.The detection limit of rhein in plasma was 16ng/ml.The recovery rate was 92.09%～98.53%.The within-day RSD was 1.65%～6.68%,and between-day RSD was 4.93%～10.01%.CONCLUSION:This method is simple and sensitive,and can be used for analysing rhein-containing biological samples.

Objective To observe the effect of total flavonoids of ampelopsis grossedentata on blood lipid and hemorheology in atherosclerosis rats .Methods The atherosclerosis model in rats was made by freeing high grease food and intraperitoneal injection of vitamin D3 ,total flavonoids of ampelopsis grossedentata was gave by gavage ,and the level of serum lipid and blood rheology were tested 24 weeks later .Results Total flavonoids of ampelopsis grossedentata could obviously decreased the level of TG ,TC ,LDL , and increased HDL(P<0 .05) .It could significantly decrease whole blood viscosity ,plasma viscosity and hematocrit (P<0 .05) . Conclusion Total flavonoids of ampelopsis grossedentata can adjust fatty substance metabolism ,improve hemorheology of athero-sclerosis rats ,and has therapeutic effect for atherosclerosis .

Objective:To study relationship among serum homocysteine (Hcy) level ,paraoxonase 1 (PON1) activity and carotid atherosclerosis .Methods:A total of 179 residents from a community of Shanghai ,who participated in cardiovascular risk factor screening from 2012 to 2014 ,were selected .They received carotid ultrasonic examination and measurements of serum Hcy ,PON1 and other biomarkers .According to serum Hcy level ,subjects were divided into elevated Hcy group (n=85) and normal Hcy group (n=94) .Results:Spearman correlation analysis indicated that serum Hcy level was significant inversely correlated with PON1 activity (r= -0.738 ,P=0.001) .Compared with normal Hcy group ,there were signifi‐cant rise in age [(60.66 ± 7.18) years vs .(64.57 ± 7.29) years] ,male proportion (27.66% vs .63.53% ) ,serum creati‐nine [(69.62 ± 12.76)μmol/L vs .(88.47 ± 20.86)μmol/L] ,uric acid [(267.85 ± 63.02)μmol/L vs .(307.51 ± 76.07)μmol/L] ,triglyceride [(1.33 ± 0.79) mmol/L vs .(1.76 ± 1.70) mmol/L]and systolic blood pressure [(134.93 ± 15.82) mmHg vs .(142.72 ± 17.86) mmHg] ,and significant reductions in levels of high density lipoprotein cholesterol [HDL‐C , (1.17 ± 0.26) mmol/L vs .(1.06 ± 0.27) mmol/L]and PON1 [(288.58 ± 73.80) kU/L vs .(187.81 ± 16.31) kU/L]in el‐evated Hcy group , P<0. 05 or <0. 01. Incidence rate of carotid atherosclerosis in elevated Hcy group was significantly higher than that of normal Hcy group (64. 7% vs .44. 7% ) , P=0. 001 .Multi‐variate gradual Logistic regression analysis indicated that serum creatinine and Hcy levels were independent risk factors for serum PON 1 activity(OR=1.055 ,1.139 , P<0.01 ,<0.05);Hcy isn′t an independent risk factor (OR=1.020 ,P=0.497) for carotid atherosclerosis .Conclusion:Serum Hcy level is significant inversely correlated with serum PON 1 activity ,and both of them are related to occurrence of carotid atherosclerosis .

Objective To investigate the expression of neuronal nitric oxide synthase(nNOS) and heme oxygenase-2(HO-2) in colon after stress in rats. Methods Water immersion-restraint stress(WRS) model was performed.The expressions of nNOS and HO-2 in colon of rats were examined by immunohistochemical staining and analysed by image analysis system. Results In control group rat,nNOS immune-positive substance mostly expressed in neurons of submucous plexus and myenteric plexus and HO-2 mostly expressed in lamina propria mucosal muscles of colon mucosa,circular muscles of colon and the endothelium and smooth muscle cells of submucosal bloodvessels.Compared with control group rat,the average gray value of nNOS positive neurons in submucous plexus and myenteric plexus and the average number of nNOS positive neurons increased significantly,the average gray value of HO-2 positive mucosal muscles and the positive unit(PU) of circular muscles increased in stress group rat,moreover,nNOS expression in mucosal epithelial cells and lamina propria lymphocytes and HO-2 expression in some large intestinal glands were observed.Compared with stress group rat,the average gray value of nNOS positive neurons in submucous plexus and myenteric plexus and the average number of nNOS positive neurons decreased in stress +L-NAME group,the average gray value of HO-2 positive mucosal muscles and the PU of circular muscles decreased in stress +ZnPP group.Conclusion Both nitric oxide(NO) and carbon monoxide(CO) are important gaseous signal molecules and neurotransmitters in colon of rats and may play a cooperative role in colonic functional disorder induced by WRS.

Gilbert Disease ; Hemochromatosis ; Humans

OBJECTIVETo compare the dissolution properties of Ginseng micropowder and common powder in vitro and investigate the effect of micronization on dissolution of Ginseng.

METHODSFive ginsenosides including Rg1, Re, Rf, Rb1, and Rb2 were determined by high performance liquid chromatography, and the dissolution curves of Ginseng micropowder and common powder were drawn.

RESULTSThe dissolution rates of ginsenosides from micropowder exceeded 90% within 5 min, significantly greater than that from common powder. But when the powders were packed in tea bags, the dissolution rates of ginsenosides were higher from common powder than from micropowder.

CONCLUSIONMicronization treatments can promote dissolution of ginsenosides from Ginseng.

Chromatography, High Pressure Liquid ; Ginsenosides ; analysis ; chemistry ; Panax ; chemistry ; Particle Size ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; chemistry ; Solubility

Objective To investigate the expressions of platelet activation-dependent granule membrane protein and platelet-derived growth factor receptor-αB, and the ultra-microstructure changes of platelets in patients with acute ST-segment elevation myocardial infarction(STEMI). Method The expressions of platelet activationdependent granule of glycoprotein (CD62P)and platelet derived growth factor receptor αβ subtype (PDGFR-αβ)of platelets in peripheral blood in 36 patients with acute ST-segment elevation myocardial infarction(STEMI) hospitalized and another 34 healthy subjects over the same period (control group) were investigated by flow cytometry and data were analyzed. The changes of ultra microstructure and activity of blood platelets in those patients and control group were observed under the scanning electron microscope. Results The expressions of CD62P and PDGFR-αβin patients with STEMI group before treatment were (3.65 ± 1.87) % and (0.43 ± 0.39) %, respectively, and those after treatment were (0.96 ± 0.79) % and (0.28 ± 0. 24) %, respectively, whereas those in control group were (0.67 ± 0.35) % and (0.27 ± 0.22) %, respectively, which were much lower in control than those in patients with STEMI before treatment (P ＜ 0.01 or P ＜ 0.05) respectively. There were statistically significant differences in the expressions of CD62P and PDGFR-αβ in patients group between pre-treatment and posttreatment (P ＜0.01 or P ＜0.05), respectively. Obvious ultra-microstructure changes of platelet surface in patients with STEMI group were observed. Conclusions Due to platelet activation in AMI, the expressions of CD62P can be used as effective indicators for monitoring coronary heart disease, and the PDGFR-αβ can be used as a reference indicator. The platelet surface ultra-microstructure changes during platelet activation in patients with AMI can be found by scanning electron microscopy.

BACKGROUNDTo establish an ADM resistant Lewis lung cancer cell line and to investigate its biological characteristics.

METHODSThe multidrug-resistant cell line was gradually induced by ADM from Lewis lung cancer cell line (L3-8), its growth characteristics and cancinogenicity were observed. The fluorescent density of ADM and Rh-B in the cells were analyzed by IPP image analysis system. The ADM concentration in cells was assayed with fluorescent meter. The IC50 was evaluated by MTT assay and the drug resistant index was counted.

RESULTSThe drug resistant Lewis lung cancer cell line, L3-8/ADM, which can grow in the medium containing 0.4mg/l ADM was acquired after 14 months selective culture. Its tumor generatic rate was 10/10. When the L3-8/ADM was cultured in 0.15mg/l ADM and general 1640 medium, the doubling time was 22.0h and 21.8h separately. After culturing in 4mg/l ADM and Rh-B medium, the fluorescent density ratio of L3-8/ADM and L3-8 were 2.82:1 and 2.65:1 separately. The ADM concentration of L3-8/ADM was only 64.7% of its parental, but there was no significant difference in their ADM release rate. Except for ADM, L3-8/ADM was resistant to DNR, VCR, MIT and CDDP in different degrees.

CONCLUSIONSL3-8/ADM cell is a typical MDR cell line. The cell membrane obstruction of drug infiltration might be the cause of drug resistance. This study will provide a basis for the establishment of lung cancer MDR animal model.

BACKGROUNDTo study the cell immunity induced by lung cancer B7 vaccine, FLB2C cells.

METHODSCompared withparental lung cancer cell LA795 line, proliferation of mouse T lymphocytes stimulated by FLB2C was observed through mixed lymphocyte culture. CTLL cell MTT test was used to detect whether FLB2C stimulated T lymphocyte to secrete IL-2. After immunized with the FLB2C and LA795 cells, the CTL activity of mouse was observed in vivo.

RESULTSThe spleen lymphocyte proliferation stimulated by FLB2C cells was remarkably stronger than that by LA795 . FLB2C might stimulated the T lymphocyte to secrete IL-2 in vitro, but LA795 didn't. FLB2C cell could induce CTL activity in vivo and the induced CTL killed FLB2C cells and LA795 in vitro, with the killing rate of 34% and 25.3% respectively; while LA795 induced CTL killing rate being 10.5% and 12.25% respectively (P < 0.01).

CONCLUSIONSFLB2C can stimulate T cell immunity in vivo or in vitro. The effect of FLB2C is significantly stronger than that of LA795 . The results suggest that FLB2C may be used to treat lung cancer through improving immunity. This provides immunological basis for applying FLB2C as a vaccine to clinical use.

Objective To compare the dissolution properties of Ginseng micropowder and common powder in vitro and investigate the effect of micronization on dissolution of Ginseng. Methods Five ginsenosides including Rg1, Re, Rf, Rb1, and Rb2 were determined by high performance liquid chromatography, and the dissolution curves of Ginseng micropowder and common powder were drawn. Results The dissolution rates of ginsenosides from micropowder exceeded 90%within 5 min, significantly greater than that from common powder. But when the powders were packed in tea bags, the dissolution rates of ginsenosides were higher from common powder than from micropowder. Conclusion Micronization treatments can promote dissolution of ginsenosides from Ginseng.