A simple thin layer chromatography ultraviolet densitometric method has been established for the separation and quantitative determination of Puerarin in plasma. Using ethyl acetate : metha-nol: water : toluene : absolute alcohol ( 100 : 15.5: 13.5: 2 :13.5 V/V ) as the developing system, 5?l supernatant of the sample extracts were spotted on high performance sillica gel G glass plates and developed. The separeted spots were scanned using Shimadzu CS-910 TLC scanner, the results were calculated by the areas of spots with Rf 0.47. The mininum detectable quantity of Puerarin was 7.5ng/spot. The plasma concentration-time curve of Puerarin in rabbits after single iv 30mg/kg was best fitted to a two- compartments open model. The T1/2? , T1/2? (3 were 5.7, 27.5 min respectively. This showed that Puerarin is distributed and eliminated in a fairly rapid rate in rabbits.

BACKGROUD: According to the record of Compendium of Herbology of herbs epimedium, Berberidaceae barrenwort genus, herbs epimedium has the effects of supplementing essence qi, consolidating bones and tendons, tonifying bruise of loin and strengthening cardiac ability, etc. In recent years, its clinicalapplications and pharmacological effects have been reported more frequently.OBJECTIVE: To investigate the effects of herba epimedium extract on spontaneous activity and sleep function of mice under the intervention of suprathreshold and subthreshold dose of pentobarbital sodium with the method of jiggle-cage transducer.SETTING: Department of Mathematics and Physics, Laboratory of Analysis, Department of Pharmacology, Gannan Medical College.MATERIALS: A total of 110 healthy adult Kunming male mice of clean grade, with a body mass between 18 g and 22 g.METHODS: The experiment was conducted in Laboratory of Scientific Research Institute of Gannan Medical College from August to September 2004. ① The mice (n=30) were injected intraperitoneally with 1 000 g/L herbs epimediu extract(0.01 mL/g) and put into hanging cage 15 minutes later, after a 3-minute stabilization period, waveforms of spontaneous activity within 15 s after the administration of drug were recorded. ②The mice (n=40) were intraperitoneally injected with 1000 g/L herba epimediu extract(0.01 mL/g), 15 minutes later, injected intraperitone ally with 0.02 mL/g suprathreshold dose of pentobarbital sodium (2.5 g/L), and then observe the disappearance time of righting reflex of the mouse. ③ The mice (n=40) were injected intraperitoneally with 1 000 g/L herba epimediu extract(0.01 mL/g), 15 minutes later,intraperitoneally injected with 0.01mL/g subthreshold dose of pentobarbital sodium (0.25%),and then observe the disappearance time of righting reflex of the mouse.MAIN OUTCOME MEASURES: Frequency of spontaneous activity of the mice; sleeping time of the mouse with suprathreshold and subthreshold dose of pentobarbital sodium.RESULTS: All the 110 mice entered result analysis. ① Herba epimedium extract (0.1 mL/10 g) could significantly inhibit the spontaneous activity of the mice, and the frequencies of large waves and moderate waves decreased obviously (7.4±6.5, 47.1±18.7; t=3.265, P ＜ 0.01). ② Herbs epimedium extract (0.01mL/10g) could shorten the time for falling asleep and prolong the sleeping time of the mouse with suprathreshold dose of pentobarbital sodium, and there was significant difference as compared with the control group [(3.9±2.8), (219.7±87.2); (5.8±2.9), (113.0±77.4) minutes; t=2.452, 2.501, P ＜ 0.05]. ③ Herba epimedium extract (0.01 mL/10 g)could significantly prolong the time for falling asleep and the sleeping time of the mouse with subthreshold dose of pentobarbital sodium, and there was significant difference as compared with the control group [(6.6±5.2),(60.3±71.7); 0, 0 minute; t=3.275, P ＜ 0.01]. Herba epimedium extract and pentobarbital sodium had synergistic effect of central inhibition.CONCLUSION: Herba epimedium extract has the effect of significant central inhibition, which could inhibit the spontaneous activity of mice ,remarkably shorten the time for falling asleep,and obviously prolong the sleeping time of the mouse with suprathreshold dose of pentobarbital sodium, prolong the time for falling asleep of the mouse with subthreshold dose of pentobarbital sodium and increase the amount of animals falling asleep.

AIM　To study the protective effect of Gn-3 (a stilbene polymer isolated from Gnetum parvifolium) against liver injury induced by CCl4, N-acetyl-p-aminophenol (APAP) and Bacillus Calmette-Guerin (BCG) plus bacterial lipopolysaccharide (LPS) in mice. METHODS　The experimental model of liver injury were induced by 0.1% CCl4 ip (10 mL*kg-1*d-1 for 3d), APAP ip (150 mg*kg-1) or BCG (5 mg) plus LPS (7.5 μg) in mice. The levels of ALT in serum, MDA and GSH in liver tissues were detected. The histopathologic changes were observed by light microscope. RESULTS　Gn-3 was shown to markedly reduce the elevated serum ALT levels, liver tissue MDA and improve the histopathological changes in all the three experimental liver injury models. No effect of Gn-3 was observed on the liver GSH level in liver injury mice. CONCLUSION　Gn-3 was found to inhibit the development of liver injury caused by CCl4, APAP, or BCG plus LPS. This means that Gn-3 has liver protective effects.

Objective: To investigate the influencing factors of depressive symptoms among rural elderly patients with chronic diseases in China, so as to provide insights into depression prevention and control among the rural elderly patients with chronic diseases. Methods: The basic demographics, health status, and lifestyle of rural residents at ages of 65 years and older who had at least one chronic disease were retrieved from The Chinese Longitudinal Healthy Longevity Survey (CLHLS) database in 2018, and participants' depressive symptoms were assessed with The Center for Epidemiological Studies Depression-10 (CES-D-10) scale. Factors affecting the depressive symptoms were identified with a multivariable logistic regression model. Results: Totally 5 146 rural elderly patients with chronic diseases were enrolled, including 2 373 men (46.11%) and 2 773 women (53.89%). The prevalence of depressive symptoms was 27.13%. Multivariable logistic regression analysis identified having two and more children (OR=0.614, 95%CI: 0.387-0.975), living alone (OR=1.450, 95%CI: 1.192-1.764), life satisfaction (general, OR=1.933, 95%CI: 1.651-2.264; low, OR=5.366, 95%CI: 3.488-8.254), self-assessed health status (general, OR=2.697, 95%CI: 2.284-3.185; poor, OR=5.338, 95%CI: 4.262-6.685), disability in instrumental activities of daily living (OR=1.592, 95%CI: 1.328-1.908), sleep duration (normal, OR=0.502, 95%CI: 0.429-0.586; too long, OR=0.494, 95%CI: 0.405-0.603), exercise (OR=0.721, 95%CI: 0.607-0.856), watching TV (OR=0.787, 95%CI: 0.664-0.933), and gardening activities (OR=0.781, 95%CI: 0.626-0.975) as factors affecting depressive symptoms among rural elderly patients with chronic diseases. Conclusions The prevalence of depressive symptoms was 27.13% among rural elderly patients with chronic diseases. Number of children, living style, life satisfaction, health status, sleep duration, exercise, watching TV, and gardening activities are associated with the development of depressive symptoms among rural elderly patients with chronic diseases.

Objective To investigate the effect of intra-articular tumor necrosis factor (TNF) inhibitor injection in patients with moderate to severe rheumatoid arthritis (RA) and values of power Doppler ultrasonography in evaluating effect of intra-articular injection.Methods RA patients with arthritis in knee and/or elbow and/or ankle referred to the Department of Rheumatology in the First Affiliated Hospital of Xi'an Jiaotong University were enrolled to receive intra-articular injection with 50 mg or 25 mg of recombinant human tumor necrosis factor-α receptor Ⅱ:IgG Fc fusion protein (TNFR:Fc) for injection after synovial fluid aspiration.Evaluation of visual analogue scale for pain of the involved joints,erythrocyte sedimentation rate (ESR),C reactive protein (CRP) and 28-joint disease activity score (DAS28) were performed before and after intra-articular TNFR:Fc injection.Synovial hypertrophy,power Doppler signal and joint effusion were analyzed and graded by ultrasound before and after intra-articular TNFR:Fc injection.Comparisons of continuous data between groups was made by t test.The data that were not normally distributed was analyzed by Mann-Whitney U rank sum test.Results Fifty-four patients with RA [6 men and 48 women,mean age (52±11) years,mean duration of disease (7±3) years] were included in this study.A significant decrease in visual analogue scale for pain of the involved joints (t=2.630,P=0.018;t=2.160,P=0.043),ESR (t=2.094,P=0.030;Z=-2.242,P=0.030),CRP (Z=-2.199,P=0.030;Z=-3.337,P=0.001) and DAS28 (t=3.579,P=0.002;t=5.538,P=0.000) were observed after one month of injection of 50 mg or 25 mg of TNFR:Fc.Synovial hypertrophy (t=2.175,P=0.036;t=2.280,P=0.030) power Doppler signal (t=2.500,P=0.020;Z=-2.504,P=0.013) and joint effusion (Z=-1.790,P=0.042;t=2.230,P=0.027) were reduced significantly after one month of intra-articular TNFR:Fc injection in knee.Synovial hypertrophy (t=2.180,P=0.034;t=2.480,P=0.030) and power Doppler signal (t=2.681,P=0.020;t=5.482,P=0.000) were also reduced significantly after one month of intra-articular TNFR:Fc injection in elbow and ankle.Conclusion Intra-articular TNFR:Fc injection is an effective and safe treatment in RA patients with monoarthritis.Ultrasound may be an objective and valid method in evaluating the effect of intraarticular TNF inhibitor injection in RA patients.

Objective To explore the distribution characteristics and function of peripheral regulatory T cells (CD4+CD25+Foxp3+T cells) in patients with systemic lupus erythematosus (SLE).In addition,we analyzed the relationship between peripheral regulatory T cells and organ damage and the influence of different treatment regimens on them.Methods Two hundred and six SLE patients and 38 healthy volunteers were enrolled,which included 12 patients with untreated new-onset lupus,11 patients with drug withdrawal more than six months and 183 patients with treatments.Phenotypic and functional analysis of peripheral blood CD4+CD25+Foxp3+T cells were performed by flow cytometry.The correlations of CD4+CD25+Foxp3+ T cells with disease activity,organ involvement were analyzed.Thealtered frequency of CD4+CD25 +Foxp3+T cells under different treatment regimens was compared.Statistical Package form Soci-science (SPSS) 21.0 software was used for data analysis,Student's t test,one-way ANOVA,Mann-Whitney T test,Kruskal-Wallis test,Chi-square test,Simple linear correlation analysis was used.Results CD4 +CD25 +Foxp3 + T cells were significantly increased inactive SLE patients [1 1.9％ (9.3％,16.0％),mean difference =104.71,P＜0.01] and inactive SLE patients [11.0％(7.7％,14.7％),mean difference=86.10,P＜0.01] compared with healthy controls [6.1％(5.3％,7.4％)].CD4+CD25+Foxp3+T cellsshowed sign-ificantly positive correlations with SLEDAI-2K (r=0.191,P＜0.05),dsDNA (r=0.262,P＜0.05),ESR (r=0.208,P＜0.05) and lgG (r=0.163,P＜0.05),and significantly negatively correlated with complementC3 (r=-0.201,P＜0.05) and C4 (r=-0.227,P＜0.05).Compared with patients without organ damage (Occult lupus),the CD4+CD25+Foxp3+T cells were increased in SLE patients with organ damage,especially those with skin involvement [10.9％(7.8％,13.1％),mean difference=56.93,P＜0.05] and renal involvement [12.1％(9.1％,16.0％),mean difference=77.26,P＜0.05].The proportion of CD4+CD25+Foxp3+T cells had no significant difference between SLE patients with treatments and patients with untreated new-onset lupus.The expressions of CTLA-4 [(53±15)％,t=7.04,P＜0.01],GITR [(42±19)％,t=2.64,P＜0.01] and ICOS [(28±9)％,t=4.27,P＜0.01] on CD4+CD25+Foxp3+T cells were significantly lower in SLE patients than in healthy controls [CTLA-4 (71±4)％,GITR (53±10)％ and ICOS (41±6)％].IL-17 synthesized by CD4+CD25+Foxp3+T cells in SLE patients [3.0％(1.8％,3.9％)] was significantly higher than that in healthy controls [1.0％(0.7％,1.2％),Z=-4.40,P＜0.01].Conclusion The peripheral regulatory T cells are significantly increased in SLE patients and correlate with disease activity and organ damage.However,their inhibitory function is defective and they have more pro-inflammatory character-istics.

Objective:To explore the distribution characteristics of memory B cells and its relationship with bone erosion in patients with rheumatoid arthritis (RA), and to further understand the mechanism of B cells in the pathogenesis of RA.Methods:B cell subsets in peripheral blood of 200 RA patients and 50 healthy individuals were detected by flow cytometry. According to the surface markers CD19, CD27 and lgD, B cells were divided into CD19 +CD27 +lgD - switched memory B cells, CD19 +CD27 +lgD + non-switched memory B cells, CD19 +CD27 -lgD - double-negative memory B cells and CD19 +CD27 -lgD + naive B cells. B cells in RA patients with various disease activity score, course of disease and treatment were analyzed. Patients were divided into four groups according to the results of joint ultrasonography, including patients without bone erosion, patients with hand bone erosion, patients with knee bone erosion and patients with hand and knee bone erosion. The relationship between the distribution of B cell subsets, autoantibodies and RA bone erosion were analyzed. Differences between the groups were analyzed by independent-samples t test, Mann-Whitney U test and χ2 test. The analysis of variance, Kruskal-Wallis analysis were used for multi-group comparison, Spearman correlation analysis was also used for correlation analysis. Results:①RA patients showed significantly decreased non-switched memory B cells [(9.5±6.7)% vs (12.1±4.7)%, t=2.46, P=0.015] and increased double negative memory B cells [(3.8±2.5)% vs(2.7±1.3)%, t=-4.74, P<0.001] in comparison to healthy individuals. The percentage of non-switched memory B cells were decreased in RA patients with moderate disease activity [(8.4±4.7 )% vs (12.4±7.5)%, t=3.13, P=0.001] and high disease activity [(7.8±7.6)% vs (12.4±7.5)%, t=3.00, P=0.003] in comparison to those in RA patients who achieved remission. Meanwhile, the na?ve B cells [(70.3±15.0)% vs (63.9±14.6)%, t=-2.15, P=0.034] were increased in RA patients with moderate disease activity. No difference was found in RA patients with different disease courses. Total B cells [(4.8±2.9)% vs (7.2±4.1)%, t=-3.24, P=0.001], non-switched memory B cells (7.6±4.3)% vs (10.0±7.1)%, t=-2.63, P=0.010) in RA patients who received prednisone treatment were decreased, while double-negative memory B cells (4.9±3.0)% vs (3.6±2.3)%, t=-2.79, P=0.006] were increased compared with those in RA patients without prednisone treatment. Non-switched memory B cells was decreased in RA patients with hand and knee erosion compared with RA patients without erosion [6.8%(2.5%, 9.5%) vs 9.7%(5.5%, 17.5%), Z=-2.12, P=0.034]. Double negative memory B cells in subgroup with keen erosion [3.3%(2.7%, 5.0%) vs 2.6%(1.9%, 3.8%), Z=-2.09, P=0.036]as well as with hand and knee erosion [3.9%(2.3%, 5.6%) vs 2.6%(1.9%, 3.8%), Z=-2.41, P=0.016] were higher than those in patients without erosion. In addition, higher serum RF level was found in subgroup RA patients with hand and knee erosion compared with subgroup of RA patients without erosion [141.0 (38.0, 874.0) U/ml vs 53.5 (10.0, 106.0)U/ml, Z=-2.07, P=0.039]. Meanwhile, the positive rate of ACPA in RA patients with bone erosion of hand was significantly higher than that of RA patients without bone erosion [81%(52/64) vs 64%(38/59), χ2=4.44, P=0.043). Conclusions:The results suggest that the increase of double negative memory B cells, the decrease of non-switched memory B cells and higher level of autoantibodies may closely relate to bone erosion of RA, which may be one of the pathogenesis of disability in RA.