This study was aimed to establish an HPLC method for the determination of chlorogenic acid, rutin and kaempferide in Solidago decurrens Lour. A Diamonsil C18 column (4.6 mm í 200 mm, 5 μm) was adopted with the mobile phase of acetonitrile-0.2% phosphoric acid. The flow rate was 1.0 mL·min-1. The column temperature was at 25℃. The wavelength of detection was set at 282 nm. The results showed that the calibration curve was linear over the range of 63.2~442.4 μg for chlorogenic acid (r = 0.999 3), 8.1~56.8 μg for rutin (r = 0.999 4) and 10.8~75.7μg for kaempferide (r = 0.999 8). The average recovery of chlorogenic acid was 98.6% (RSD = 1.4%), that of rutin was 99.2% (RSD = 0.8%) and that of kaempferide was 100.3% (RSD = 1.0%). It was concluded that the method was simple, economical and accurate with good reproducibility.

Objective To investigate the oxidative status in systemic sclerosis (SSe) and explore the correlation between oxidative status and clinical features by measuring serum 8-isoprostaglandin-F2α (8-isoPGF2α) concentrations. Methods Serum 8-iso-PGF2α levels were detected by enzyme-linked immunosorbent assay (ELISA) in 51 SSe patients and 22 matched healthy controls. And in some SSc patients, serum homocysteic acid (Hey) concentrations were examined by cyclophorasc assay. Plasma yon Wilebrand factor (vWF) activity and serum immunoglobulin (lg) concentrations were detected by immunoturbidimetry.Antinuclear antibodies (ANA) and anti-endothelial antibodies (AECA) were detected by indirect immunofluorescence. Anti-sc170 antibody was detected by immunoblotting. Thirty-five out of 51 SSc patients were assessed for clinical features and laboratory parameters in order to analyze the correlations between 8-isoPGF2α levels and clinical features. Results Serum 8-iso-PGF2α levels were higher in sclerederma patients than in healthy controls. Values of 8-iso-PGF2α correlated with pulmonary involvement, such as diffusion capacity for carbon monoxide (DLCO) and pulmonary interstitial fibrosis by pulmonary high-resolution computed tomography (PhrCT), and correlated positively with renal vascular damage determined by the resistant index (RI) of renal glomeruli interlobular arteries. There was no correlation between 8-iso-PGF2αconcentrations and skin, peripheral vascular, heart, esophagus involvement and disease activity, diseasepattern, vWF,Hcy, lg, or autoantibedy profiles.Conclusion Increased Jevels of 8-iso-PGF2α, marker of oxidative stress, is correlated with pulmonary fibrosis and the extent of renal vascular damage in SSc and this supports the hypothesis that oxidative stress plays an important role in SSc pathogenesis.

This study was aimed to establish the fingerprints of Qian Solidaginis Herba by HPLC. The fingerprints of Qian Solidaginis Herba were built by using Diamonsil C18 (200 mm í 4.6 mm, 5 μm) column and acetonitrile -0.2%H3PO4 aqueous solution in gradient as mobile phase. The flow rate was 1.0 mL·min-1. Detecting wavelength was set at 282 nm. The total 10 batches of Qian Solidaginis Herba samples and their adulterants were analyzed. The results showed that 13 peaks were identified as the fingerprints of Qian Solidaginis Herba. Under chromatographic conditions mentioned above. It was concluded that the HPLC fingerprints method was found to have satisfactory accuracy, sta-bility and reproducibility, which was a potential method for the quality control of Qian Solidaginis Herba.

AIM: To establish HPLC fingerprint for the identification of Hydrocotyle sibthorpioides and its adulterants. METHODS: The chromatographic seperation was performed on a Diamonsil C_(18) (250 mm×4.6 mm,5 μm)with a mobile phase consisting of acetonitrile-0.1% phosphoric acid, gradient eluent, at the flow rate of 1.0 mL/min. The UV detection was set at 340 nm. RESULTS: There was an apparent difference in fingerprint between Hydrocotyle sibthorpioides and its adulterants. CONCLUSION: The method is stable and reliable with a good reproducibility andprovides a reference standard for identifying Hydrocotyle sibthorpioides and its adulterants.

Objective To identify interleukin 17 (IL-17) and B cell activating factor (BAFF) that could influence B cell biology by detecting the expression of BAFF in the serum and labial salivary glands from primary Sj(o)gren's syndrome (pSS) patients and to test the apoptosis rates of B cells cultured with Th17 cells which were transfected with IL-17-siRNA,BAFF-siRNA.Methods A total of 40 patients with pSS who were referred to the Department of Rheumatology and Immunology at Anhui Provincial Hospital from June 2011 to June 2012 were enrolled into this study.The expression of BAFF on salivary gland and serum from pSS patients and healthy controls were detected by ELISA and immunohistochemical examination (22 patients with pSS).Flow cytometry was used to detect B cell's apoptosis,BAFF and IL-17 interfered with amplified Th17 cells,and co-cultured with B cells.Immunoblot was used to detect supernatant antibody in 5 patients with pSS.Independent samples t test was used for statistical analysis.Results In all pSS specimens,infiltrating inflammatory cells expressed BAFF,so did some ductual cells,but acinar cells did not express these markers.There was no expression of BAFF in the controls.BAFF-positive cell numbers in the labial salivary glands of pSS patients with focal infiltrating lymphocytes were more than that with non-focal infiltrating lymphocytes (888±372 vs 164±161,t=5.94,P＜0.05),and the percentage of BAFF-positive lymphocytes over the total infiltrating lymphocytes in the salivary glands of pSS patients with focal infiltrating lymphocytes [(0.18 ±0.08) ％] was higher than those with non-focal infiltrating lymphocytes [(0.09 ±0.07) ％] (t =3.03,P＜0.05).The level of soluble BAFF in patients with pSS [(6.0±2.8) ng/ml] was significantly higher than the controls [(3.8±1.7) ng/ml,t=3.26,P＜0.05].BAFF or IL-17 transfected group,B cell apoptosis rate [(24± 5)％,(23±5)％] were significantly higher than the non-transfected group [(7±4)％],t=4.6,4.4; P＜0.05].And there was no significant difference when compared with cultured B cells (P＞0.05).Compared with the controls,no antibody could be detected in the supernatants.Conclusion BAFF may be involved in the process of local inflammatory damage of the pSS,it may have a synergistic effect with IL-17 on abnormal B cell function.

Objective To investigate the changes of plasmacytoid dendritic cells (pDC) in peripheral blood of patients with systemic lupus erythematosus(SLE) and its roles in the pathogenesis of SLE.Methods The level of pDC and the expressions of CD32,CD40,CD86,CD62L and CXCR4 were analyzed by flow cytometry.The concentrations of IFN-α in serum were detected by ELISA assay.Results The levels of circulating pDC were significantly decreased in SLE patients compared with healthy controls.Moreover,the pDC levels in active SLE patients were lower than those in inactive SLE patients,and compared with the primary group,the pDC levels were increased in the treatment group.The levels of pDC showed a significant decrease in SLE patients with arthritis,proteinuria or leucopenia in comparison with patients without those manifestations,showing a negative correlation with proteinuria.The expressions of cell surface molecules including CD32,CD86,CD62L and CXCR4 on pDCs were significantly increased in SLE patients compared with healthy controls,and the levels of pDC were negatively correlated with the expressions of CD32 and CXCR4 in patients with SLE.The concentrations of IFN-α in serum of patients with SLE were significantly higher than those in healthy controls,and the levels of pDC were positively correlated with the concentrations of IFN-α in patients with SLE.Conclusion The level of circulating pDC in patients with SLE was remarkably reduced,but the expressions of molecules involved in cell activation and migration were upregulated,accompanied by enhanced IFN-α production,which might promote the onset and progression of SLE.

Objective To measure the number of peripheral blood CD34+ hematopoietic stem/progenitor cells (HSC/HPCs) and membrane expression of CD34 on these cells in patients with SLE. Methods Lymphocytes were isolated from peripheral blood of 30 patients with SLE and 14 normal human controls. Flow cytometry using FITC-labeled antibodies was performed to determine the percentage of CD34+ HSC/HPCs and mean fluorescence intensity (MFI) of CD34 on these cells. Their correlation with clinical data was analyzed.Results The percentage of CD34+ HSC/HPCs in peripheral lymphocytes was (0.15 ± 0.10)% and (0.09 ±0.07)% in active and stable SLE patients, respectively, significantly lower than that in normal controls [(0.37 +0.17)%, F = 17.18, P ＜ 0.01], however, there was no significant difference between active and stable SLE patients (t = 1.51, P＞ 0.05). The MFI of CD34 was higher in active SLE patients than in the normal controls (41.35 ± 19.24 vs. 27.43 ± 7.57, F= 3.13, P ＜ 0.05), but no difference was observed between stable SLE patients and normal controls (F= 3.13, P ＞ 0.05). In patients with SLE, the percentage of CD34+ HSC/HPCs was negatively correlated with serum IgG levels (r = -0.588, P ＜ 0.01 ), but uncorrelated with SLE disease activity index (SLEDAI) or serum levels of complement, anti-dsDNA antibodies, anti-C1q antibodies, antinucleosome antibodies, etc. Conclusions The count of CD34+ HSC/HPCs is reduced while the MFI of CD34 antigen is elevated in SLE patients, hinting that there is a functional abnormality of HSC/HPCs in SLE patients, which may be involved in the pathogenesis of SLE.

Objective To measure the number of peripheral blood CD34+ hematopoietic stem/progenitor cells (HSC/HPC) expression of CD34 in the peripheral blood of patients with rheumatoid arthritis and exploreits relationship with clinical manifestations. Methods CD34+ HSC/HPCs in the peripheral blood of RA patients (n=32) and healthy controls (n=16) were detected using flow cytometry. The relationship between the frequency of HSC/HPCs, mean fluorescence intensities (MFI) of CD34 and clinical manifestations and rheumatoid factor (RF), anti-cyclic citrullinated peptide (CCP) antibodies, disease activity score (DAS) 28,X rays stages and healthy assessment questionnaire (HAQ) were analyzed. Student's t-test and pearont test were used for statistical analysis. Results Frequency of CD34+ HSC/HPC in the peripheral blood of RA patients was decreased compared with normal controls [ (0.13±0.09)% vs (0.38±0.21)%, P＜0.05 ], CD34 MFIwere higher in RA patients than those in the normal controls (57±33 vs 3111, P＜0.05). The frequency was positively correlated with the number of (RBC red blood cell), (Hb hemoglobin), and was negatively correlated with C-reactive protein (CRP), and the MFI of RA patients was positively correlated with healthy assessment questionnaire (HAQ) and X ray stages, but negatively correlated with the number of platelets.Conclusion CD34+ HSC/HPC of the peripheral blood of RA patients are significantly abnormal, which is characterized by decreased CD34+ hematopoietic stem cell, and the decrease is positively correlated with RBC and Hb, but negatively correlated with CRP. CD34+ hematopoietic stem cell may play an important role in the pathogenesis of RA.

OBJECTIVE:To establish the HPLC fingerprint for Blumea balsamifera and its fake B. riparia. METHODS:HPLC was performed on the column of Uitimate-C18 with mobile phase of acetonitrile-0.05% phosphoric acid(gradient elution)at a flow rate of 0.6 mL/min,detection wavelength was 270 nm,column temperature was 25 ℃,and injection volume was 7 μL. Using quercetin as a reference,Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicine(2004 A edition)was used for the common peaks identification and similarity analysis of 16 batches of B. balsamifera and 5 batches of B. ri-paria. RESULTS:There were 61 common peaks in the 16 batches of B. balsamifera,similarity degree was 0.931-0.995,which was higher than the similarity degree of 5 batches of B. riparia. CONCLUSIONS:The established fingerprint can provide reference for the identification and quality evaluation of B. balsamifera.

AIM: To establish HPLC fingerprint for the identification of Hydrocotyle sibthorpioides and its adulterants. METHODS: The chromatographic seperation was performed on a Diamonsil C_18(250 mm?4.6 mm,5 ?m)with a mobile phase consisting of acetonitrile-0.1% phosphoric acid,gradient eluent,at the flow rate of 1.0 mL/min.The UV detection was set at 340 nm. RESULTS: There was an apparent difference in fingerprint between Hydrocotyle sibthorpioides and its adulterants. CONCLUSION: The method is stable and reliable with a good reproducibility and provides a reference standard for identifying Hydrocotyle sibthorpioides and its adulterants.