Objective To establish a HPLC method for the determination of psoralen and bergapten in Ficus hirta Vahl.Methods A Merck-lichrospher C18(4 mm?250 mm,5 ?m)column was adopted.The mobile phase was acetonitrile-water(35∶65) with the flow rate of 1.0 mL/min.The column temperature was 35 ℃,and the detection wavelength was set at 222 nm.Results Psoralen's linearity was obtained in the range of 0.12 ?g～1.20 ?g(r=0.999 7),and bergapten's linearity in the range of 0.03 ?g～0.30 ?g(r=0.999 5).Psoralen's average recovery was 100.9 %with RSD 2.9 %,and that of bergapten was 99.6 %with RSD 1.9 %.Conclusion This is the first report of simultaneous determination of psoralen and bergapten in Ficus hirta Vahl.by HPLC,and the results showed the method is accurate,reproducible,and can serve as quality evaluation method for Ficus hirta Vahl.

BACKGROUND:Because macrophages play an important role in the body’s inflammatory response, the detection of the impact of biological materials on the behavior of macrophages can assess the immunogenicity of materials. OBJECTIVE: To analyze the activation effect of decelularized extracelular matrix materials on macrophages. METHODS: The peritoneal macrophages of BALB/c mouse were obtained and cultured by dividing into five groups. Control group was simple cel culture group, experimental group 1 was acelular matrix membrane material directly contacting with macrophage for culture, experimental group 2 was fresh pericardial material directly contacting with the macrophage for culture, experimental group 3 was acelular matrix membrane material indirectly contacting with macrophages for culture, experimental group 4 was fresh pericardium material indirectly contacting with macrophages for culture. After 24 hours of culture, the secretion of nitric oxide and cytokines in cel culture supernatant was determined. After 48 hours of culture, the absorbance value was determined by MTT method and the toxicity grading was determined. RESULTS AND CONCLUSION: The toxicity grading in experimental groups 1-4 was respectively grades 2, 4, 0, 2. The nitric oxide level in experimental groups 1 and 2 was higher than that in the control group (P < 0.05), and the nitric oxide level in the experimental group 2 was higher than that in the experimental group 1 (P < 0.05). There were no significant differences in interleukin-2, interleukin-4,interferon γ, interleukin-17A and interleukin-10 levels between these five groups. The interleukin-6 level in the experimental group 2 was higher than that in the control group (P < 0.05); The expression levels of tumor necrosis factors in experimental group 1, 2 and 4 were higher than those in the control group (P < 0.05), and experimental group 2 higher than the experimental group 1 (P < 0.05), experimental group 1 higher than the experimental group 4 (P < 0.05). These results show that acelular matrix material can activate macrophages in direct contact.

Objective To investigate the clinical and prognosis characteristics of SLE patients whose initial clinical presentations were Sj(o)gren's syndrome (SS).Methods Medical charts of 41 consecutive SS/SLE inpatients admitted to Peking Union Medical College Hospital (PUMCH) from February 1998 to February 2008 were systematically reviewed,including demographic data,clinical features,laboratory findings,treatment as well as prognosis.Two hundred and fourteen cases were randomly selected as controls from 2331 non SS-onset lupus (NSs/SLE) inpatients weated in PUMCH at the same time period.Results There were significant differences between SS/SLE and NSS/SLE patients in the following aspects (P＜0.05):①gender composition:(F/M) (41/0 vs 184/30),age at the diagnosis of SLE [(43±41) yrs vs (32±31) yrs)],disease duration L(114±84) mollths vs (45±18) months];②clinical features:xerostomia (85.3% vs 6.1%),xerophthalmia (75.6% vs 2.3%),faeial rash (9.8% vs 46.3%),renal tubular acidosis (21.9% vs 0),nephrotic syndrome (7.3% vs 31.3%),central nervous system invoivement (4.9% vs 19.6%),interstitial lung disease (12.2% vs 2.8%);③labratory findings:ESR[(65±75) mm/1 h vs (46±34) mm/1 h)],patients with elevated IgG level (56.4% vs 29.9%) and IgA level (38.5% vs 20.4%),RF,prevalence of anti-SSA and anti-SSB antibodies (70.8% vs 20.3%,82.9% vs 43.4% and 39.0% vs 7.9%);④SLEDAI score (8±8 vs 10±10),glucocorticosteroid treatment (methylprednisolone bolus/1～2 mg·kg-1·d-1 prednisone/1 mg·kg-1·d-1 prednisone)(8/26/7 vs 91/102/21),and rate of death and/or severe irreversible organ failure (2.4% vs 14.9%).SS/SLE patients were followed up for (33±34) months,40 cases remained stable and only one patient died of acute pulmonary embolism.Conclusion Compared to NSS/SLE,SS/SLE patients ale older and have more insidious disease course.They have higher prevalence rate of anti-SSA and anti-SSB antibodies,renal tubular acidosis and interstitial lung disease,but less severe neuropsychiatric and renal involvements and much better prognosis.

Objective To investigate the pharmacodynamics of different local anesthetics administered intrathecally for elderly patients undergoing transurethral resection of the prostate (TURP). Methods Ninety ASA Ⅰ - Ⅲ elderly patients, aged 69-82 yr, with body mass index less than 30 kg/m2 , undergoing TURP under combined spinal-epidural anesthesia, were randomly divided into 3 groups ( n = 30 each): levobupivacaine group (group L), ropivacaine group (group R) and bupivacaine group (group B). Group L, R and B received intrathecai (IT) 0.5 % levobupivacaine, 0.5 % ropivacaine and 0.5 % bupivacaine respectively. The initial dose was 7,10 and 6 mg in group L, R and B respectively. The ratio of two successive doses was 0.9. If the upper sensory block reached T10 within the 20 min after IT injection, the IT analgesia was considered to be effective. The median effective dose (EDs0) and 95 % confidence interval (95 % CI) were calculated by Dixon. Results The ED50 and 95% CI of levobupivacaine, ropivacaine and bupivacaine were 6.781 (95% CI 6.561-7.024) mg, 9.135 (95%CI8.670-9.616) mg and 5.170 (95% CI 5.012-5.333) ng respectively. The relative potency ratio between levobupivacaine, ropivacaine and bupivacaine is 0.76∶0.57∶1.00. ConclusionThe relative potency ratio be tween levobupivacaine, ropivacaine and bupivacaine is 0.76∶0.57∶1.00.

Objective To study the relationship between graft-versus-host disease (GVHD) and endothelium injury following hematopoietic stem cells transplantation in mice. Methods C57BL/6 mice as donors and Balb/c mice as recipients were randomly divided into 4 groups: control group, bone marrow transplantation group, GVHD group, GVHD mitigation group. The clinical manifestations,circulating endothelial cells and tissue pathological changes were observed at different time points after transplantation. Results No manifestations of GVHD were found in each group at the day 5, while those were found in GVHD group at the day 9 and all died within 15 days. The counts of endothelial cells in peripheral blood showed no significant difference at the day 5 between GVHD group (7. 34 ±1.26 cells/μl) and bone marrow transplantation group (11.51 ± 7. 40 cells/μl) or GVHD mitigation group (7. 36 ± 0. 16 cells/μl), while among three groups there was statistically significant difference at the day 9 (GVHD group: 153. 64 ± 35. 35 cells/μl vs bone marrow transplantation group: 10. 49 ±5. 61 cells/μl and GVHD mitigation group: 47. 82 ± 4. 69 cells/μl). The scores of pathological aGVHD had no significant difference at the day 5 between GVHD group (4. 33± 1. 53) and bone marrow transplantation group (3. 33 ± 0. 58) or GVHD mitigation group (4. 00 ± 1.73), while among three groups there was statistically significant difference at the day 9 (GVHD group: 10. 0 vs bone marrow transplantation group: 3. 33 ± 1.15 or GVHD mitigation group: 4. 33 ± 0. 58) and at the day 14 (GVHD group: 10. 33 ± 2. 58 vs bone marrow transplantation group: 2. 33 ± 1.25 or GVHD mitigation group 3. 33 ± 1.15). Conclusion Occurrence of GVHD causes endothelial damage again and injured endothelium worsens the GVHD.

Objective To investigate the curative effect of granulated cancellous bone autograft in treatment of refractory bone nonunion after limited contouring of bone ends. Methods Between 2003 and 2006, 13 patients with refractory bone nonunion were treated with external fixation and granulated cancellous bone autograft after limited contouring of bone ends. Results The mean follow-up period was 22.6 months (19-30 months), which showed that all patients gained bone union and resumed com-plete weight loading or previous job at final follow-up. The mean fixation time of external fixators was 10.6 months (7-18 months). The intermittent or persistent pin-track infection occurred in eight patients and relieved by pin-track care and oral or parenteral antibiotics, with no infection after removing external fixator. Conclusion The granulated cancellous bone autograft after limited contouring of bone ends is an effective method for treatment of refractory bone nonunion.

Purpose The diagnosis, treatment and prognosis of benign and malignant biliary stricture are significantly different. This study aims to evaluate the quantitative analysis of biliary structures using magnetic resonance cholangiopancreatography (MRCP) combined with dynamic contrast enhanced CT (DCE-CT).Materials and Methods The quantitative parameters of MRCP and DCE-CT imaging data from 27 patients with benign biliary stricture (benign group) and 30 patients with malignant biliary stricture (malignant group) were retrospectively analyzed. The wall thickness, stricture length and diameter, proximal ductal dilatation and degree of enhancement in two groups were compared, and its correlation was analyzed to evaluate the accuracy of MRCP and DCE-CT.Results There were significant differences in wall thickness [(3.2±2.0) mmvs (2.1±0.6) mm], stricture length [(15.8±8.1) mmvs (9.5±6.5) mm] and diameter [0 mmvs (2.0±0.9) mm], proximal ductal dilatation and the degree of enhancement [(12.7±3.6) mmvs (9.3±2.7) mm] between the two groups (t=2.825, 3.270, 4.025,P<0.001;Z=-3.909,P<0.001). Multivariable stepwise regression analysis showed that the wall thickness and diameter, and the CT HU in portal venous and equilibrium phases combined with CT plain scanning were significant predictors of malignant biliary strictures (t=-6.424-2.309,P<0.05). The sensitivity, specificity, inter-modality agreement and Youden index of MRCP and DCE-CT in diagnosing 57 patients with biliary stricture were 96.67%, 100.00%, 98.25% and 0.97, respectively; with statistical significance in predicting benign and malignant biliary stricture (AUC=0.994,P<0.001).Conclusion Using MRCP and DCE-CT, the wall thickness and diameter of the stricture, and the difference in CT HU in portal venous and equilibrium phases combined with CT plain scanning are valuable in differential diagnosis of benign and malignant biliary stricture.

Accurate determination of HIV-1 proviral burden and viral load is very useful in prognosis of HIV-1 infected patients and in assessment of drug for therapy of AIDS patients. In order to establish a quantitative method in detecting HIV-1 proviral burden and viral load, 8E5 cell line and a recombinant RNA constructs were used as the HIV-1 proviral DNA and viral RNA external references, respectively. The PCR products were labeled with the fluorescent DNA dye SYBR green. The amount of burden or load was measured by GeneAmp 5700 Sequence Detection System. Using this method, the HIV-1 proviral burdens in PBMC of patient and in cell suspension treated with the compounds AZT, GL and WT were measured. HIV-1 viral loads in supernatant of the cell culture treated with the above compounds were also determined. The therapeutic indices (TIs) of the compounds calculated based on the inhibition of virus induced syncytial formation, and inhibitionn of proviral burdens and viral loads were compared, and their TIs successively increased. The fluorescent real time quantitative PCR possesses very good specificity, sensitivity and duplication. TI value of a drug based on inhibition of proviral burden in cell culture, and the proviral burden in PBMC of patient may be useful in evaluating a drug on eradicating provirus from resting and memory CD4 T cells.

AIM:To investigate the effect of mesalazine treatment on regulation of Th1, Th17 and Treg cells in mice with dextran sulfate sodium ( DSS)-induced ulcerative colitis ( UC) .METHODS:The expression of IL-17, IFN-gam-ma and Foxp3 in the peripheral blood mononuclear cells ( PBMC) and intestinal mucosa lamina propria mononuclear cells ( LPMC) of DSS-induced UC mice was detected by flow cytometry analysis.The effect of mesalazine treatment on regulaiton of Th1, Th17 and Treg cells in the mice with DSS-induced ulcerative colitis was examined.RESULTS:The expression of IL-17, IFN-γand Foxp3 on CD4 +T cells were significantly higher in the PBMC of DSS-induced mice than those in control group.CD4+IFN-γ+T cells and CD4 +Foxp3 +T cells were higher in LPMC than those in control group, except CD4+IL-17 +T cells.Moreover, the Th1, Th17 and Treg cells were higher in DSS group than those in control group in LPMC.How-ever, only Tregs was higher in PBMC.Pre-treatment with mesalazine significantly decreased the number of Th17, Th1 and Treg cells of UC model mice both in PBMC and LPMC.CONCLUSION: The Th1, Th17 and Tregs cells in DSS-induced mice were significantly higher than those in control mice, suggesting that CD4 +T cell subsets play an important role in the pathogenesis of UC.Mesalazine may play a role in the treatment of UC by regulating the Th1, Th17 and Tregs cells.

Objective To (e)xplore inhibition of endothelial progenitor cells (EPCs) against hepatic vein thrombosis after allogeneic bone marrow transplantation (BMT).Methods Balb/c mice were randomly divided into three groups: (1) BMT group [Balb/c mice were injected intravenously with 5 × 106 bone marrow cells after total body irradiation (TBI)]; (2) EPCs co-transfusion with bone marrow cells group: 5 × 105 EPCs were infused into recipient mice simultaneously; (3) Normal control group.Liver index was detected on the day 0,5,10,15 and 20 after transplantation.Hepatic vein thrombosis,hepatic cells and vascular endothelial damage were observed under the light microscopy after H&E staining.The injury of liver cells,liver veins,hepatic sinusoidal endothelial cells (SECs)and platelet adhesion conditions were observed under a transmission electron microscope (TEM).The proportion of activated platelets and TNF-α concentration in peripheral blood were detected by using flow cytometry.Results On the day 0,5,10,15 and 20 after transplantation,the proportion of activated platelets,liver index and TNF-α concentrations in BMT group and EPCs co-transfusion group showed an upward trend,peaked on the 15th day,and then decreased.However,they were still significantly higher than those in normal control group (P＜0.05).The above parameters in EPCs co-transfusion group at each time point were significantly lower than those in BMT group (P＜0.05).As compared with BMT group,platelet adhesion decreased,hepatic vein thromboses were reduced,hepatocyte swelling and necrosis were alleviated,and liver damage repaired rapidly in EPCs co-transfusion group.Conclusion EPCs co-transfusion with bone marrow cells could inhibit the hepatic veins thrombosis and ameliorate liver damage significantly.