Methods The maximum tolerated dose( MTD)of water and ethanol extracts of Radix millettiae speciosae were measured for the safety evaluation. Results Maximal tolerated dose( MTD)of water extract was higher than 1 000 g·kg-1 ,which equaled to 110 times the dose for human adults(60 kg). MTD of ethanol extract was higher than 1 700 g·kg-1 ,which equaled to 186 times the dose for human adults(60 kg). Conclusion Radix millettiae speciosae does not have obvious toxicity,thus its routine clinical dose is safe and feasible.

Objective To explore the role of hydroxychloroquine (HCQ) in ultraviolet B (UVB)- induced expression of interleukin (IL)-10 and interferon (IFN)-γ from CD4+T cells in patients with systemic lupus erythematosus (SLE). Methods Thirty patients with SLE and 10 healthy controls were enrolled in the study. CD4+ T cells were isolated using magnetic beads from SLE patients and healthy controls. HCQ was added in culture media before and after irradiation with UVB 311 nm narrow band ultraviolet B (NB-UVB). The levels of IL-10 and IFN-γ in the supernatant were detected with enzyme-linked immunosorbent (ELISA). Comparisons between groups were performed by t-test. Results The level of IL-10 was higher in SLE patients [(27±4) pg/ml] than that in healthy controls [(18±3) pg/ml, P=0.011]. After exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages, the level of IL-10 was increased significantly in patients with active disease (P=0.022, P=0.048). After exposure of CD4+T cells to UVB in 100 mJ/cm2 dosages, the levels of IL-10 was higher in patients with active disease [(77±42) pg/ml] than patients with stable disease [(24± 4) pg/ml, P=0.029]. When CD4+ T cell were cultured with HCQ, IL-10 and IFN-γ levels in patients with active disease [(2.6±4.0), (17.5±2.3) pg/ml] were decreased significantly (P=0.018, P=0.017). HCQ reversed UVB-induced IL-10 expression in active SLE patients after exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages (P=0.037, P=0.04). HCQ also reversed UVB-induced IFN-7 expression in active SLE patients and stable SLE patients after exposure to CD4+T cells with UVB in 100 mJ/cm2 dosages (P=0.013, P= 0.049). Conclusion UVB can aggravate the imbalance of Th1 and Th2 cytokines. HCQ inhibits UVB-induced IL-10 and IFN-7 expression of CD4+T cells in patients with SLE, especially in patients with active disease.

Objective To explore the effect of hydroxychloroquine on ultraviolet ray-induced genomic DNA hypomethylation in CD4+ T cells from patients with systemic lupus erythematosus (SLE). Methods Thirty patients with SLE and 10 normal human controls were enrolled in the study. CD4+ T cells were isolated from these subjects by using magnetic beads, and cultured. Hydroxychloroquine of 50 mg/L was added to the culture medium of CD4+ T cells before or after the exposure to narrow band ultraviolet B (NB-UVB) 311 nm.After additional culture, the levels of genomic DNA methylation in CD4+ T cells were determined with the Imprint Methylated DNA Quantification kit. Results The levels of DNA methylation was lower in SLE patients than in the normal controls [(3.922 ±2.215)% vs. (10.210 ± 5.573)%, t= 3.450, P = 0.026]. After exposure to UVB at 45 and 100 mJ/cm2, the DNA methylation level in patients with active SLE decreased from (7.235 ±3.846)% to (1.784 ± 1.033)% and (1.932 ± 1.844)% respectively (t= 3.000, 4.118, both P< 0.05). Decreased DNA methylation level was observed in CD4+ T cells from patients with active SLE compared with those from patients with stable SLE and normal human controls [(1.932 ± 1.844)% vs. (7.235 ± 3.846)% and (5.472 ±5.573)%, t = 2.648, 3.000, both P< 0.05] after irradiation with UVB of 100 mj/cm2. A significant increase in the methylation level was observed in active SLE patient-derived CD4+ T cells treated with hydroxychloroquine following the irradiation with UVB of 45 (4.698% ± 1.948%) and 100 mJ/cm2(8.698% ± 3.151%) compared with those only treated with UVB irradiation (t = 4.827, 3.184, both P< 0.05), as well as in those treated with hydroxychloroquine before and after the irradiation with UVB of 45 mJ/cm2 compared with those receiving irradiation alone [(5.404 ± 2.308)% vs. (1.784 ± 1.033)%, t = 4.827, P< 0.01]. Conclusion Hydroxychloroquine can reverse the UVB-induced genomic DNA hypomethylation in CD4+ T cells from patients with SLE,especially in those from patients with active SLE.

Objective To study the role of peripheral blood T regulatory cells (Treg) markers, as well as different Treg cells in the pathogenesis of systemic lupus erythematosus (SLE) and explore the correla-tion betweenCD127 and Foxp3. The immunosuppressive effect of CD4~+CD25~+CD127~(low/-) T cell is explored. Methods ①Four-color direct fluorescence-labeled and multi-parameter flow cytometry were used to detect peripheral blood CD4~+CD25~+ T cells and other Treg cells accounted for the proportion of CD4~+ T cells in 40 SLE patients (19 cases with active disease 21 cases in remission) and 15 healthy controls. Meanwhile, its correlations with anti-dsDNA antibodies among 7 groups were analyzed.②Flow cytometry sorting combined with cell culture were applied to detect and analyze the proliferation inhibition effect of CD4~+CD25~+CD127~(low/-)regulatory T cells to CD4~+CD25~-effector T cell.Two independent samples t test,ANOVA for repeated measures,Pearson's correlation and Spearman's correlation were used for statistical analysis.Results ①The cell ratio of the 7 groups of SLE patients was(6.1±1.7)%,(3.1±1.3)%,(2.1±1.0)%,(1.6±0.3)%(0.97±O.28)%,(0.69±0.23)%and(O.71±0.35)%respectively.In the SLE group,the proportion of the first 6 groups was lower than the control group (P<0.05).For CD4~+CD127~(low/-)Foxp3~+,there wag no difference in the two groups(P>O.05).②CD4~+CD25~+Foxp3~+,CD4~+CD25~(high)Foxp3~+ T cells and IgA ratio Was positivelv correlated.While CD4~+CD25~(high)CD127~(low/-) T cells and anti-SSB antibodies was positively correlated in SLE patients.③The seven group of cells,in addition to CD4~+CDl27~(low/-)Foxp3~+ T cell ratio were lower in early-onset SLE patients than those in patients at remission(P<0.05).④The cell propor-tion of the first 6 groups was lower than that of post steroid treatment(P<0.05).⑤Foxp3 expression of CD4~+CD25~+T ceils and CD4~+CD25~(high) T cells was positively correlated with low expression of CDl27 in the early onset group,remission group and the control group.⑥The CD4~+CD25~- effect T cells could be suppressed by their own CD4~+CD25~+CDl27~(low/-) regulatonry T cells in vitro,and the inhibition of SLE patients was significantly lower than controls.Conclusion The immunological abnormality of SLE may be associated with the qnatity and functional defects of immune regulatory T cells.CDl27 may be a possible alternative to Foxp3 regulatory T cell-specific surface markers.

Objective To investigate the expression of regulatory T cells in the peripheral blood of rheumatoid arthritis patients and to explore the possibility of using CD127 instead of FOXP3 in the study of regulatory T cells. Methods Thirty-two patients with active rheumatoid arthritis (RA) [ including 20 patients who hadn't been treated with disease modifying antirheumatic drugs (DMARDs) and 12 patients whose disease were poorly improved after DMARDs therapy], 25 patients with primary Sjogren's syndrome (pSS) and 24 healthy controls were selected. The percentages of CD4+CD25high, CD4+CD25+CD127low/'- and CD4+CD25+ FOXP3+T cells were analyzed by 3-color flow cytometry. Meanwhile, the levels of CRP, ESR, immunoglobulin and complement were measured. Results Percentages of CD4+CD25+CD127low/-T cells and CD4+CD25+FOXP3+ T cells in RA patients were significantly lower than those of healthy controls (P<0.01 for each category of T cells),but the percentage of CD4+CD25high T cells didn't show significant changes (P>0.05). The expression of CD4+CD25+CD127ow/- and CD4+CD25+FOXP3+T cells showed no difference between RA and pSS patients (P> 0.05 for each). Differences of the three group cells between untreated RA patients and poorly-improved RA patients were not significant (P>0.05 for each case). CD4+CD25+CD127low/-T cells were positively associated with CD4+CD25+FOXP3+T cells (r=0.698, P=0.001), but the two groups had no correlation with the levels of CRP, ESR and anti-CCP antibody and RF (P>0.05 for each case). Conclusion The percentage of CD4+CD25+CD127low/-T cells is decreased in the peripheral blood of active RA patients and positively correlates with CD4+CD25+FOXP3+T cells.It suggests that CD127 may be an ahemative marker of FOXP3 in the study of Treg cells.

Objective To study the expressions of Toll-like receptor 9 protein (TLR9) in peripheral B and T lymphocytes in newly diagnosed, untreated patients with systemic lupus erythematosus (SLE) and their relationship with clinical parameters. Methods Blood samples were obtained from 35 newly diag-nosed, untreated patients with SLE and 16 healthy human controls. B, T lymphocytes and TLR9 protein were labeled with fluorescent antibodies, and the expressions of TLR9 protein were detected by flow cytometry in peripheral B and T lymphocytes. The relationship between TLR9 expression and clinical parameters was assessed. Results The proportions of B and T lymphocytes expressing TLR9 in newly diagnosed, untreated patients were (53.94±17.95)% and (49.33 ± 23.30)%, respectively, compared to (29.40 ± 10.54)% and (29.18 ± 14.78)%, respectively, in healthy controls (t = 6.11,3.73, respectively, both P < 0.01). Additionally,the proportion of B lymphocytes expressing TLR9 correlated negatively with SLE disease activity index (SLEDAI)(r = -0.39, P < 0.05), but positively with the level of serum IgA antibody (r = 0.74, P < 0.01).Condnsions The expression of TLR9 is elevated in peripheral T and B lymphocytes from patients with newly diagnosed, untreated SLE, and the proportion of TLR9-expressing B lymphocytes negatively correlates with SLEDAI, but positively correlates with the serum level of IgA antibody.

Objective To evaluate the safety of Solanumprocumbens by studying its acute toxicity to mice. Methods The dosage of 100% death ( Dm) and 100% survive ( Dn) were determined. Five groups were set between the dosage of Dm and Dn in a 1:0.8 ratio, and then were intragastrically administrated once at the dosage of 250,200,160,128,102.4 g·kg-1 respectively.Toxicity and mortality of mice after intragastricly administration of Solanumprocumbens were observed for 14 days continuously. Results After four hours of administration, there were death in each group except the lowest dosage group (102.4 g·kg-1).Number of death of the groups 250,200,160 and 128 g·kg-1 were 10,8,6 and 3 respectively.LD50 of Solanumprocumbens was 153. 02 g · kg-1 , the 95% confidence interval was ( 136. 55, 171. 47 ) g · kg-1 . Conclusion Solanumprocumbens has a certain toxicity.More attention should be payed to its toxicity for clinical rational drug use.

Objective: To study the causes and influencing factors of no seeking medical treatment among pneumoconiosis patients. Methods: Using stratified sampling method and typical survey method, we carried out the questionnaire survey in nine provinces in China including the east, the medium and the western region using a questionnaire on the seeking medical behavior pf pneumoconiosis patients and the influencing factors. The subjects include occupational pneumoconiosis cases and clinically diagnosed cases and the response rate is 94.3 percent. The data was entered twice with epidate3.1 and error detection and statistical analysis was completed with SPSS 20.0. Chi-square test was used for univariate analysis, and multivariate logistic regression was used for multivariate analysis. Results: One thousand and thirty-seven subjects were investigated with average age 55.9±11.2 years. Seventy percent of them were silicosis and 21.9 percent were coal worker’s pneumoconiosis with 67.5 percent of them residing permanently in the countryside, and 37.9 percent of their education background were primary school culture and 33.1 percent of them had junior high school culture. Thirty two point six percent of respondents had no personal income with a median monthly income of 1 200 yuan. Four hundred and thirty four of subjects hadn't seek medical treatment since they got the pneumoconiosis accounting for 41.9 percent with three hundred and thirty seven of them hospitalized directly. The reasons of no seeking medical treatment for the respondents mainly include the self-induction symptoms lighter, the high cost of treatment and cannot claiming the payment of the medical expenses, buying drugs in drugstore, thinking that no medicine can cure pneumoconiosis or no effect, complex procedures, too far away from medical institutions, no unaccompanied, needing a long time or no time, communication disorders, etc. accounting for 44.4 percent, 24.6 percent, 10.9 percent, 9.1 percent, 6.9 percent, 4.4percent, 3.2 percent, 2.9 percent, 1.9 percent, 1.5 percent, respectively. The results of multivariate analysis showed the main characteristics of subjects with restrictions to the outpatient health service utilization are as follows: demographic sociological indicators such as registered permanent residence area is western (OR_western=2.18, 95%CI:1.38-3.43) , more than seventy five years old (OR_{over 75}=6.82, 95%CI:2.04-22.9) , unemployment, temporary or permanent employment (OR _unemployment=1.90, 95%CI:1.17-3.08; OR_{temporary employment}=3.11, 95%CI:1.57-6.14; OR_{permanent employment}=2.10, 95%CI:1.18-3.74) , self-rated health score of 50 or above (OR_{self-rated-70}=2.04, 95%CI:1.18-3.51; OR_{self-rated-90}=3.00, 95%CI:1.97-5.37; OR_{self-rated 90}=2.95, 95%CI:1.74-8.07) ; with increase to the outpatient health service utilization are breath with difficulty (OR=0.57, 95%CI:0.41-0.78) , emphysema (OR=0.48, 95%CI:0.26-0.90) , hospitalized with pneumo-coniosis (OR=0.12, 95%CI:0.07-0.20) . Conclusion Pneumoconiosis patients no covered by injury insurance should be orderly included in the basic medical security system, and be given the medical treatment actively; It should be strengthened the health management for the pneumoconiosis patients and correctly guided the utilization of medical services.

OBJECTIVE To investigate the anti-inflammatory effects and mechanism of Zhuang medicine Tongfeng li’an capsules on gouty arthritis in combination with in vivo and in vitro experiments. METHODS Sixty rats were randomly divided into normal group， model group， positive control group （27 mg/kg allopurinol+0.27 mg/kg colchicine）， Tongfeng li’an capsules low- dose， medium-dose and high-dose groups （2.2， 4.5， 9.0 g/kg）， with 10 rats in each group. Except for normal group， gouty arthritis model of rats was induced in other groups. Rats in each administration group were given corresponding drugs intragastrically， and rats in the normal group and model group were given equal volume of water intragastrically for 14 consecutive days. The degree of ankle joint swelling， serum level of interleukin-1β （IL-1β） and protein expressions of nuclear factor kappa-B （NF-κB） in synovial tissue were detected， and the histopathological changes of synovium tissue in the ankle joint of rats were observed. The inflammation model was established by stimulating RAW264.7 cells with lipopolysaccharide. After Tongfeng li’an capsules （62.5， 125， 250 μg/mL） were given， the levels of nitric oxide （NO）， reactive oxygen species （ROS） and IL-1β in the cells and protein expression of NF-κB were detected， and NF-κB localization in the cells was also determined. RESULTS Results of in vivo experiment showed that compared with normal group， the swelling degree of the ankle joint， serum IL-1β level and protein expression of NF-κB in synovium tissue were all increased significantly in model group （P＜0.05）； pathological changes such as synovial hyperplasia， edema， vascular congestion， capillary hyperplasia， and increased inflammatory cells were observed. Compared with model group， the levels of above indexes were all decreased significantly in Tongfeng li’an capsules high-dose group （P＜0.05）， and most of the above indexes were significantly reduced in Tongfeng li’an capsules medium-dose and low-dose groups （P＜0.05）； synovial hyperplasia of the ankle joint improved， and the infiltration of inflammatory cells 2019BS044） decreased. Results of in vitro experiment showed that Tongfeng li’an capsule could significantly reduce the levels of NO， ROS and IL-1β and protein expression of NF-κB（P＜0.01）， and inhibit NF- κB nucleation. CONCLUSIONS Tongfeng li’ancapsules have good anti-inflammatory effect on gouty arthritis， and its mechanism may be related to the inhibition of NF-κB signaling pathway activity.

Aldehyde dehydrogenase 2 (ALDH2) is one of important factors against from the damage under oxidative stress in human body. A high proportion of East Asians carry ALDH2 inactive mutation gene. There are many diseases closely related to ALDH2, such as cardiovascular diseases, neurodegenerative diseases and liver diseases. Recent studies also have found that ALDH2 is associated with ferroptosis. Therefore, ALDH2 has becoming a potential target for the treatment of the above related diseases. Several types of small molecule activators with potential value of clinical application have been reported. The research progress on the structure and function of ALDH2 , the relationship with human diseases and its activators were summarized in this paper.