OBJECTIVE:To determine the content of paeonol in Cynanchum paniculatum by three-dimensional fluorescence coupled with ATLD algorithm. METHODS:The trilinear model was established. The principle of least square was used for ATLD. The component value was fitted by core consistency diagnosis,and inner filter effect was corrected by mathematical correction method. Fluorescence scanning condition included excitation wavelength of 250-400 nm,emission wavelength of 410-600 nm, wavelength interval of 5 nm,slit width of 5.0/5.0 nm,scanning speed of 1200 nm/min. Determination and absorption spectrum condition included scanning wavelength of 250-600 nm,scanning speed of 600 nm/min;Al(Ⅲ)was used as the sensitizer to in-crease the fluorescence intensity of paeonol. The content of paeonol was determined by HPLC. RESULTS:The linear range of pae-onol was 0.132-1.188 μg/mL(r=0.9999). RSDs of precision,stability and reproducibility tests were less than 3.0%. The recover-ies were 100.1%-104.7%(RSD=2.39%,n=6). The analysis spectrum of paeonol was almost completely overlapped with the actu-al spectrum. The results of HPLC method are very similar to those of ATLD algorithm. CONCLUSIONS:The three-dimensional fluorescence coupled with ATLD algorithm is simple,rapid,efficient and accurate,and can be used for the qualitative and quantita-tive analysis of complex system.

Objective To investigate glucose metabolism , insulin secretion and serum lipid in patients of nonalcoholic fatty liver disease (NAFLD) with hypertension. Methods A total of 100 subjects including NAFLD with hypertension (n = 50) and NAFLD without hypertension (n = 50), in our hospital during Jul. 2010 to Jul. 2012, were enrolled in this study. Records the following information of the two groups: The clinical data , oral glucose tolerance and insulin releasing test , serum lipid , serum uric acid and ALT were recorded. Results Compared with the NAFLD without hypertension group, impaired glucose regulation was significantly increased in patients in the NAFLD with hypertension group(P < 0.05).The insulin releasing was significantly increased at 30 min and was increased at 180 min in patients in the NAFLD with hypertension group (P < 0.05).The insulin releasing during 30 min to 120 min was descend gradually in patients in the NAFLD with hypertension group,with no significant difference between these two groups. TG was markedly elevated in patients in the NAFLD with hypertension group (P < 0.05). Conclusion Prevalence of impaired glucose regulation was elevated obviously in NAFLD with hypertension group,and early-phase insulin secretion were reduced and delayed.

Objective To examine the correlation between urotensin Ⅱ (UⅡ) concentration and the severity of liver fibrosis. Methods Liver fibrosis model was induced in rats by intraperitoneal administration of CCl4. At 4,6,8 weeks, the rats were sacrificed, and the hepatic tissue hydroxyproline (HYP) content was measured. Hepatic tissue specimens were histopathologically evaluated according to a fibrosis scoring system. Plasma UⅡ levels were measured by ELISA method. Results Plasma UⅡ gradually increased with the increase of duration of CCL4, UⅡ concentration correlated to liver fibrosis ( R2 = 0.875, P ＜ 0.05) and hepatic HYP( R2 = 0.65, P ＜0.05). Conclusion UⅡ was involved in the pathogenesis of liver fibrosis.

Objective To analyze the false-positive results of Treponema pallidum antibody caused by 3 different assay in comparison with Treponema pallidum hemagglutination assay (TPHA).Methods Research group included 3957 clinically asymptomatic syphilis patients,and control group was 344 outpatients with sex-transmitted diseases (STD).The serum samples from the patients who were TPHA-positive were tested in parallel by enzymeimmunoassay (EIA) and syphilis toluidine red untreated serum test (TRUST).Western blot (WB) was performed as confirmatory test.Results In the clinically asymptomatic patients,60 were TPHA-positive.Among them 57 were confirmed by western blot assay,and 1 was false-positive and 2 were borderline in WB.Of the 60 TPHA-positive patients,53 were positive in EIA and 23 were positive in TRUST.In STD patients 40 were TPHA,WB and EIA-positive but 32 were TRUST-positive.Conclusions The results of TPHA and EIA were consistent for diagnosis of syphilis patients who may suffer from previous or latent infection.

AIM: To develop the methods for the quantitative analysis of gallic acid and total phenolic acid in Sedum Aizoon L. METHODS: Gallic acid was analyzed by HPLC on a Hypersil BDS C_(18) column and detected at 271 nm.The mobile phase was methol-water(adjusted to pH=3.0 with H_3PO_4)(90∶10)at a flow rate of 1.0 mL/min.Total phenolic acid was analyzed by spectrophotometry at 720 nm.The colour-developing agent was the mixture solution of 0.6%FeCl_3—0.9%K_3[Fe(CN)_6](1∶0.9). RESULTS: Calibration graphs were constructed in the range of 0.343 0-1.200 ?g(r=0.999 7) for gallic acid and 0.4640-2.320 ?g/mL(r=(0.999 3)) for total phenolic acid.The average recoveries were 97.91%(n=6) for gallic acid and 99.21%(n=6) for total phenolic acid.The RSD of the methods were 1.8% and 2.0%,respectively. CONCLUSION: The methods were fast,reliable,accurate and suitable for analysis of gallic acid and total phenolic acid and quality control in Sedum Aizoon L.

Forty-five male mice were used and divided into three groups: i.e. experimental gastric ulcer group, saline control group and normal control group. After experimental gastric ulcer was induced, the mice of three groups received intraperitoneal injection of colchicine and were sacrificed 3h later at 3, 6, 9 and 20 days, respectively. The antral mucosa was removed and processed by Sternberger's immunocytochemical PAP method to show G cells and counterstained with hematoxylin. In normal control group, the mitotic index of the antral mucosal epithelial and glandular cells was 5.93?1.23; the percentage of G cells was 2.76?0.45; the mitotic index I of G cells (the number of the mitotic G cells per 100 G cells) was 0.85?0.18; and the mitotic index II of G cells (the number of mitotic figures of the G cells per 100 antral epithelial, glandular and G cells) was 0.02?0.01. The mitotic index of the antral mucosal epithelial and glandular cells, the percentage and the mitotic index II of G cells on 6th, 9th and 20th days in experimental gastric ulcer group was raised and showed highly significant statistical difference from that of the control group, respectively. The mitotic index I of G cells was raised on 9th day in the experimental gastric ulcer group and significant difference between experimental gastric ulcer group and the control group was found. It also revealed a significant diference in the experimental gastric ulcer group as compared with saline control group on 20th day. The percentage of G cells on 6th day was most high, but the peak of mitotic number of G cells appeared on 9th day in the experimental gastric ulcer group. The distribution of G cells was found upward in the glands near the ulcer on 3rd and 6th day than in normal control. These findings suggest that the number, origin, distribution and shape of the G cells in the pyloric glands exhibited dynamic changes with the passage of time. The results suggested that the G cells might participate in the regulation of regeneration of antral mucosa during experimental gastric ulcer.

AIM: To establish HPLC fingerprint of Sishen Pill and a determination of main component of Sishen Pills(Fructus Evodiae, Semen Myristicae, Fructus Psoraleae, etc.) by HPLC. METHODS: The samples were extracted with the solvent methanol. The separation was performed on YWG-C 18(4.6mm?250mm, 10?m) column with water- methanol-acetonitrile as a mobile phase gradient elution. The flow rate was 1.0mL?min -1, the column temperature was at 30℃ and the wavelength of UV detector was at 225nm. RESULTS: There are 21 higher peaks and 12 marker peaks in the fingerprint of eight batches of samples. For evodiamine, the regression equation was Y=147.37X+1186.32, r=0.9999, the liner range was from 5 to 1000?g?mL -1 and the recovery was 99.83%. For rutaecarpin, the regression equation was Y=101.19X+279.81, r= 0.9999, the liner range was from 4.5 to 900?g?mL -1. The recovery was 98.72%. CONCLUSION: The quality standard of Sishen Pill was established firstly. It can be used for quality cantrol of Sishen Pill.

ObjectiveTo investigate whether hypothalamic silencing of suppressor of cytokine signaling 3( SOCS3 ) by RNA interference ( RNAi ) attenuates diet-induced obesity and leptin resistance in rats.Methods Hypothalamic SOCS3-deficient rats were established by means of lentiviral vector ( LV ) mediated RNAi technique,and then were fed with a high-fat diet for 8 weeks.After the rats were sacrificed,the serum leptin and insulin concentrations were measuredbyRIA, andthe expressionof SOCS3inhypothalamus was detectedby immunohistochemistry and realtime PCR.ResultsThe immunostaining showed that SOCS3 protein expression was significantly reduced and the expression of SOCS3 mRNA was decreased by 49％ in SOCS3 RNAi group compared to the controls( P＜0.01 ).The rats with hypothalamic SOCS3 knockdown exhibited a significant hampering in gaining body weight with lowered concentrations of leptin[ ( 8.18±2.10 vs 10.85±2.23 ) ng/ml ],insulin[ ( 18.89±4.88 vs 26.78±6.01 )mU/L],glucose [ (4.89±0.91 vs 6.26± 1.41 )mmol/L] and triglyceride [ (0.47 ±0.10 vs 0.62 ±0.16)mmol/L] when exposed to a high-fat diet(P＜0.05 or P＜0.01 ).ConclusionThe results provide evidence that rats with hypothalamic SOCS3 silencing by RNAi are resistant to diet-induced obesity,leptin resistance,and metabolic disorder.

Objective To explore the mechanism of glucocorticoids in the treatment of acute lung injury(ALI) by studying the expression changes of aquaporin-1 (AQP-1) in the lung.Methods SpragueDawley rats were randomly divided into 3 groups,which were ALI group injected by lipopolysaccharide (LPS group),therapy group treated by methylprednisolone (Mp) after LPS injection (Mp group) and control group injected by normal saline.PaO2 and wet/dry mass ratio were assayed at 6 hours after the injection.Changes of AQP-1 expression in the lungs were analyzed by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR),and pathological examination was performed in each group.Results RT-PCR results showed that the expression of AQP-1 in LPS group was lower than that in control group (0.28 ± 0.05 vs 0.50 ± 0.24,P ＜ 0.05),while the expression of AQP-1 in Mp group was higher than that in LPS group (0.47 ±0.16 vs 0.28 ±0.05,P ＜0.05).The immunohistochemistry results were consistent to the RT-PCR results,meanwhile glucocorticoids therapy could improve the blood oxygen level (P ＜ 0.01),reduce the wet/dry mass ratio (P ＜ 0.05) and relieve the lung injury.Conclusion Glucocorticoids may have beneficial effects on ALI by affecting the expression of AQP-1 in the lungs of rats.

Objective To investigate the changes of immune function,serum procalcitonin (PCT) and C-reactive protein (CRP) in children with mycoplasma pneumoniae pneumonia (MPP),and to provide clinical evidence for immunotherapy in children with MPP.Methods A total of 126 children with MPP during their hospitalization were enrolled into lobar pneumonia group (n =42) and bronchopneumonia group (n =84),and 28 healthy children were enrolled into normal control group.The immunoglobulin(Ig),CD4+T,CD8+T,PCT and CRP of all children were determined.Results The levels of IgG,IgM and IgE of children in lobar pneumonia group and bronchopneumonia group were significantly higher than that of the normal control group (P ＜0.05),but there were no significant differences in the level of IgA between three groups (P ＞ 0.05).The levels of IgG in lobar pneumonia group were significantly higher than that of bronchopneumonia group (P ＜ 0.05),but there were no significant differences in the levels of IgM,IgE,IgA between the two groups(P ＞0.05).The ratios of CD4+T and CD4+T/CD8+T in lobar pneumonia group and bronchopneumonia group were significantly lower than that of the normal control group(P ＜ 0.05).The levels of PCT and CRP in lobar pneumonia group and bronchopneumonia group were significantly higher than that of the normal control group (P ＜ 0.05).Conclusion Humoral immunity and cellular immune dysfunction plays an important role in pathogenesis of MPP.They are important for PCT and CRP in evaluating clinical condition and immunotherapy.