Objective To optimize the extraction process of Euphorbia hirta L.by Central composite design/response surface methodology with ethanol by the ultrasonic extraction technology.Methods Based on the single factor experiment and central composite design-response surface methodology,the content of quercitroside was regarded as evaluation index,select ethanol concentration,extraction time,and solid-liquid ratio as three evaluation factors to achieve the purpose. Results The results indicated that the optimal extraction process was obtained as follows:10 times the amount of 60% ethanol under ultrasound for 2 times,50 min each;the average content of the quercitroside was 2.39 mg/g.Conclusion The method to optimize the extraction process is simple and reasonable.

Objective To evaluate the effect of enteral nutrition emulsion on the immunologic function and intestinal mucous barrier in diabetic patients. Methods Eighty diabetic patients were randomly divided into con-trol group (n=40) and enteral nutrition group (n=40). The urine lactulose (L) and mannitol (M) levels and the blood immunologic indicators recorded on day 1 and day 8. Results The L/M ratio was significantly lower in enteral nutrition group than in control group on day 1 and day 8 ( P < 0. 05 ). The IgG level was significantly higher in enteral nutrition group than in control group on day 8 ( P = 0. 02 ). Conclusion Enteral nutrition emulsion can decrease the permeability of intestinal mucous membrane and improve the immunologic function in diabetic pa-tients.

OBJECTIVE:To establish a method for simultaneous determination of the contents of ligustroflavone,specnuezhe-nide,demethylwedelolactone and wedelolactone in Zibu ganshen pill. METHODS:Dual-wavelength HPLC was performed on the column of Elite C18 with mobile phase of acetonitrile-0.5%acetic acid(gradient elution)at flow rate of 0.9 ml/min,column temper-ature was 25 ℃,detection wavelengths were 224 nm(0-30 min) and 351 nm (30-50 min),and the injection volume was 20 μl. RESULTS:The linear range was 6.75-135.00μg/ml(r=0.999 5)for ligustroflavone,6.54-130.80μg/ml(r=0.999 8)for specnuezhe-nide,4.90-98.00μg/ml(r=0.999 4)for demethylwedelolactone and 6.42-128.40μg/ml(r=0.999 6)for wedelolactone;RSDs of pre-cision,stability and reproducibility tests were no more than 1.25%;average recoveries were 96.15%-99.96%(RSD<2%,n=6). CONCLUSIONS:The method is rapid,sensitive and accurate,and can be used for the contents determination of ligustroflavone, specnuezhenide,demethylwedelolactone and wedelolactone in Zibu ganshen pill.

Objective To study the risk factors of Pseudomonas aeruginosa (PA)infection and homology of PA isolated from nasopharyngeal carcinoma patients undergoing radiotherapy,and prevent and control the spread of in-fection.Methods Bacteria isolated from clinical specimens were identified by BD Phoenix automated microbiology system. Gene homology were analyzed with randomly amplified polymorphic DNA (RAPD)technique.Results Forty-nine strains of PA were mainly isolated from 43 nasopharyngeal carcinoma patients,the major specimens were nasopharyngeal swab (46.94% ),sputum(32.65% ),and oral secretion (10.20% ).All these strains were amplified 46 electrophore-sis diagrams,and 19 genotypes were identified. The highly homologous genotypes of type H and J strains were mainly isolated from patients in the second section (57.14% ,4/7)and fourth section (60.00% ,3/5)of radiotherapy department respectively. Conclusion Localized epidemic of highly homologous PA exists in different sections of ward,transfer of patients between different sections is the risk factor for homology PA infection/colonization. Gen-otyping technique such as RAPD for analyzing the homology of pathogenic bacteria in healthcare-associated infection has important value in preventing and control Ling infection spread.

Objective:To develop an HPLC-DAD-ELSD method for the determination of nitidine chloride, 5-ethoxychelerythrine, bergeninum and ardisiacrispin A in Shangtong tinctures ( STD) . Methods: A Hypersil C18 column was used as the chromatographic column, the flow rate was 0. 8 ml·min-1 . For nitidine chloride and 5-ethoxychelerythrine, the mobile phase A consisted of acetoni-trile,the mobile phase B consisted of 0. 1% formic acid-triethylamine (pH 4. 5),and the DAD detection wavelength was at 273 nm. For bergeninum and ardisiacrispin A, the mobile phase consisted of methanol-water(25∶75), the temperature of drift tube was set at 95℃, and the gas flow (N2) was set at 2. 5 SLPM·min-1. Results:There was a good linear relationship between the concentration and peak area for nitidine chloride and 5-ethoxychelerythrine within the range of 0. 021-0. 426 μg (r=0. 999 5) and 0. 075-1. 494 μg (r=0. 999 8), respectively. The average recovery was 99. 22%(RSD=0. 64%) and 98. 61%(RSD=0. 46%), respectively. There was a good linear relationship between the concentration and peak area for bergeninum and ardisiacrispin A within the range of 0. 215-4. 304 μg(r=0. 999 3) and 0. 286-5. 728 μg(r=0. 999 7), respectively. The average recovery was 99. 15%(RSD=0. 77%) and 99. 25%(RSD=0. 56%) accordingly. Conclusion:The method is accurate, sensitive and reproducible, and can be used in the de-termination of nitidine chloride, 5-ethoxychelerythrine, bergeninum and ardisiacrispin A in STD.

BACKGROUND: Revascularization is a challenge for the tissue-engineered bone carrying cells after implanted into human body. Previous studies have found that tanshinol can improve the functions of endothelial progenitor cells and exert vascular protective effects.OBJECTIVE: To prepare the β-calcium phosphate (β-TCP) scaffold with tanshinol coating, and to observe its cytocompatibility.METHODS: The β-TCP scaffolds coated with 10-7, 10-6 and 10-5 mol of tanshinol were constructed by negative pressure absorption method. The distribution of tanshinol coating on the scaffold was observed using scanning electron microscopy,and the inner ingredients were analyzed by infrared spectrum. Human endothelial progenitor cells (hEPCs) were cultured in the extracts of β-TCP and β-TCP scaffolds with 10-7, 10-6, 10-5 mol of tanshinol coatings, respectively. The cell proliferation was detected at 2, 4, 6, 8 and 10 days of culture; the levels of nitric oxide and vascular endothelial growth factor in the supernatants were detected at 1, 7 and 14 days of culture; the lumen formation on the matrigel was observed after 14-day culture. hEPCs were respectively seeded onto the β-TCP and β-TCP scaffolds with different dosages of tanshinol coating,and then the cell growth was observed under scanning electron microscope at 7 days.RESULTS AND CONCLUSION: The tanshinol coating evenly distributed on the inner surface of the pores, and its crystalline structure became dense with dosage increasing. Infrared spectrum analysis revealed no changes in the characteristic absorption peak of tanshinol and TCP in the scaffold. The β-TCP scaffolds with tanshinol coating could promote the proliferation of hEPCs, especially the scaffolds with 10-6 and 10-5 mol tanshinol coating. Compared with the β-TCP scaffold, the scaffolds with 10-6 and 10-5 mol tanshinol coating significantly upregulated the nitric oxide level at 14 days of culture, and significantly increased the level of vascular endothelial growth factor at 7 and 14 days of culture (P <0.05). Although it could be found in all β-TCP scaffolds with tanshinol coating, the lumen formation was the maturest in the scaffold with 10-5 mol tanshinol coating. These results suggest the β-TCP scaffolds with tanshinol coating can promote the proliferation and endothelial differentiation of hEPCs, and hold a good cytocompatibility.

Objective To explore the etiopathogenisis of fungal infections induced by systemic lupus erythematosus (SLE). Methods One hundred and forty-seven SLE patients with fungal infections during 2004-2013 were assigned as experimental group and the same number of SLE patients without infections were randomly selected as control group. Clinical and laboratory documents of these patients were comparatively analyzed. Results The fungal infections in SLE patients affected oral mucosa , lower respiratory tract and skin , and the pathogenic bacteria were mainly candida albicans. The wide application of glucocorticoid, immunosuppressants and antibiotics pushed the rise of incidence of fungal infections in SLE patients significantly. Conclusions The patients′ use of glucocorticoid , immunosuppressants and antibiotics attributes to a higher risk of fungal infections , especially infections by candida albicans. Fungus infection may raise the level of C4.

Objective To investigate the effect of long-term low dose prednisone administration on bone mineral density (BMD) in patients with inactive systemic lupus erythematosus (SLE).Methods A total of 118 inactive female SLE patients with long-term administration of low dose prednisone were recruited from the Department of Rheumatology and Immunology at An hui Provincial Hospital.All patients were given low dose prednisone for long-term (≤ 10 mg/d,more than half a year).According to prednisone doses,subjects were divided into two groups,namely group A (≤7.5 mg/d) and group B (7.5-10 mg/d).In addition,patients were also divided into four groups based on the duration of administration,including group Ⅰ ≤3 years,Ⅱ from 4-5 years,Ⅲ 6-10 years and Ⅳ ＞ 10 years.Twenty-nine healthy people were recruitedas normal controls.The BMD was measured by dual energy X-ray absorptiometry.The association of BMD with prednisone dose and duration was compared between different groups.Results The incidencc of osteopenia in all patients with SLE was 42.4％ (50/118),and the incidence of osteoporosis was 14.4％ (17/118).BMD of all bone sites in both group A and B were significantly lower than that in normal control group (P ＜ 0.05).Similarly,the BMD of all bone sites in group Ⅰ,Ⅱ,Ⅲ and Ⅳ were significantly decreased (P ＜ 0.05).What needed to be stressed was the BMD in group Ⅳ was lower than those in other three groups (P ＜ 0.05).Multiple logistic regression analysis showed that the cumulative prednisone dose was the risk factor for osteopenia,while taking calcium and alfacalcidol were protective factors.Conclusion Long-term use of low dose prednisone result in the decrease of BMD in patients with inactive SLE.The lumbar spine and femoral neck had more severe osteopenia.Long-term administration of prednisone,even less than 7.5 mg/d,can also cause osteopenia.Calcium and alfacalcidol were protective factors of BMD.

Objective To analyze the expression of purinergic receptor P2X ligand-gated ion channel 7 (P2X7R) on different cells and peripheral blood mononuclear cell (PBMC) and to investigate its correlation with inflammatory cytokines in patients with SLE.Methods Flow cytometry was used to detect surface expression of P2X7R on lymphocytes,CD4+ cells,and CD19+ cell in 29 SLE patients and 28 healthy human controls to compare the difference between the SLE patients and the controls in P2X7R expression.Enzyme linked immunosorbent assay (ELISA) was performed to detect P2X7R-related serum cytokines interleukin (IL)-1β,IL-6,tumor necrosis factor (TNF)-α level.T test,Wilcoxon rank sum test,Spearman's correlation analysis were used for statitical analysis.Results ① SLE patients had significantly higher expression of P2X7R on CD4+,CD8+ lymphocytes compared to controls [CD4+ cells∶ 2.21(3.55) vs 0.89(1.15),Z=-1.527,P=0.015； CD19+ cells∶ 11.53(20.01) vs 6.66 (6.27),Z=-2.091,P=0.037]； ② The levels of three cytokines in patients with SLE were significantly higher than those in control.The positive relationship between P2X7R expression in lymphocytes with the serum IL-6 level was found in SLE patients (r=0.449,P=0.015)；③ Patients with arthritis showed significantly higher expression of P2X7R on lym-phocytes compared to patients without arthritis (Z=-2.772,P=0.006).The expression of P2X7R on lymphocytes and CD19+ cell was significantly positively correlated with the SLEDAI score.Positive correlation with anti-β2GP Ⅰ in lymphocyteswas also found.Conclusion P2X7R may mediate the release of inflammatory cytokines involved in the pathogenesis of SLE,and may participate the development of arthritis,lupus nephritis and NPSLE in SLE patients.

Objective To detect the levels of peptidylarginine deiminase 4 (PAD4) mRNA and neutrophil extracellular traps (NETs) of the peripheral blood neutrophils in rheumatoid arthritis (RA) patients and to study the association between PAD4 and NETs in RA.The anti-cyclic citrullinated peptide (anti CCP) antibodies,disease activity score (DAS28) score,erythrocyte sedimentation rate (ESR) were recorded.Its value in the pathogenesis of RA was explored.T test,rank-sum test,Pearson and Spearman correlative analysis were used for statical analysis.Methods The serum double-stranded DNA (dsDNA)/NETs in 43 RA patients and 20 healthy controls were detected by PicoGreen dsDNA Quantitation Kits (Invitrogen).Real-time quantitative polymerase chain reaction (RT-PCR) was used to determine the expression of PAD4 mRNA in neutrophils,and the relationship between dsDNA/NETs and their clinical indexes were analyzed.Results ① The level of peripheral blood neutrophils PAD4 mRNA in RA was significantly higher than healthy controls [1.493(0.831,2.607)] vs [0.631(0.358,1.489)],(Z=-2.07,P=0.039).The levels of serum PAD4 mRNA in RA treated with disease-modifying antirheumatic drugs (DMARDs) were lower than untreated with DMARDs but no significant difference was found (Z=-1.19,P=0.234).② The levels of serum dsDNA/NETs in RA patients were significantly higher than in healthy controls (t=3.4,P=0.001).③ The levels of serum dsDNA/NETs in RA were positively correlated with DAS28 score,ESR,CRP and PAD4 mRNA (r=0.36,P=0.036;r=0.345,P=0.042;r=0.36,P=0.043;r=0.42,P=0.017) but not with anti-CCP antibodies (r=0.277,P=0.154).Conclusion ① PAD4 may play an important role in the pathogenesis of RA.② NETs maybe associated with disease activity and play an important role in RA pathogenesis,inhibit NETs generation or improve the ability to clear NETs,perhaps can treat RA.③ PAD4 probably play an important role in rheumatoid arthritis by influencing the formation of the NETs.