Objective To evaluate the efficacy of Yishen-Tongdu decoction combined with sulfasalazine and meloxicamin for ankylosing spondylitis(AS).Methods A total of 86 AS patients were recruited and randomized into an observation group and a control group,43 in each group.The control group was treated by sulphasalazine and meloxicam,while the observation group was treated by Yishen-Tongdu decoction combined with sulfasalazine and meloxicam.The physical function and the disease activity were measured with the Bath Ankylosing Spondylitis Functional Index (BASFI) and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI),respectively.The erythrocyte sedimentation rate (ESR) and serum C-reactive protein (CRP) were determined.Results The BASFI (3.25 ± 1.18 vs.4.18 ± 0.96;t=4.544,P＜0.01) and BASDAI (2.33 ± 1.46 vs.3.26 ± 1.43;t=5.245,P＜0.01) after treatment in the observation group were significantly decreased than those in the control group.Serum CRP level (8.62 ± 14.71 mg/L vs.12.57 ± 16.32 mg/L;t=3.143,P＜0.05) and ESR (14.93 ± 17.15 mm/h vs.18.61 ± 20.98 mm/h;t=3.615,P＜0.05) in the observation group were significantly lower than those in the control group.The occiput-wall distance (2.07 ± 0.59 vs.2.68 ±0.69 cm;t=5.332,P＜0.01),finger-floor distance (12.88 ± 1.92 vs.13.26 ± 1.71 cm;t=3.593,P＜0.05),and jaw-sternum distance (1.58 ± 0.63 vs.2.43 ± 0.64 cm;t=4.671,P＜0.01) in the observation group were significantly decreased than those in the control group,while the chest expansion (4.99 ± 0.73 cm vs.4.26 ± 0.68 cm;t=4.226,P＜0.01),and Schober test (6.57 ± 0.91 vs.6.13 ± 0.87em;t=3.733,P＜0.01) in the observation group were significantly increased than those in the control group.Conclusion Yishen-Tongdu decoction combined with sulfasalazine and meloxicam can improve the physical function and the disease activity and tts effect is superior to sulfasalazine and meloxicam in patients with AS.

Objective:To investigate the effect of Glycyrrhizin on the cytokines derived from peritoneal macrophages in mice.Methods:Glycyrrhizin was intraperitoneally administered 24 hr before the peritoneal macrophages (PMs) were harvested.The harvested PMs were then stimulated in vitro with lipopolysaccharide (LPS).The levels of tumor necrosis factor (TNF)-?,interleukin-12 (IL-12) p70,interleukin-10 (IL-10) and macrophage colony-stimulating factor (M-CSF) from culture supernatants were measured by ELISA.Results:Glycyrrhizin suppressed LPS-induced TNF-? production and increased LPS-induced IL-12 p70 production by PMs significantly. The production of IL-10 and M-CSF by PMs were not effected by Glycyrrhizin pretreatment.Conclusion:These findings demonstrate the ability of Glycyrrhizin to suppress LPS-induced TNF-? poduction and to enhance IL-12 production by peritoneal macrophages.

Objective: To study effect of small ubiquitin related modifier protein 1 (SUMO1) in inflammatory reactions mediated by tumor necrosis factor (TNF)-α and nuclear factor (NF)-κB in myocardial damage of rats with type 2 diabetes mellitus (T2DM). Methods: A total of 20 Goto-Kakizaki (GK) rats with spontaneous diabetes mellitus (DM) were randomly divided into group DM1 (pure DM group, n=10) and group DM2 (DM+high-fat diet group, n=10), and another 10 normal Wistar rats were regard as healthy control group. Expressions of SUMO1, TNF-α and NF-κB were measured by immunohistochemical method. Results: 1. Levels of blood glucose and TG in group DM1 and group DM2 were significantly higher than those of healthy control group, and those of DM2 group were higher than of DM1 group ,P<0.05 all; 2. Myocardial cells lined up in order and there was no hypertrophy in group DM1; but those in group DM2 showed cells loosely lined up and hypertrophy under light microscope; 3 Immunohistochemical assay indicated that expression of SUMO1 in group DM2 and DM1 group were significantly higher than those of healthy control group [(44.5±1.1) vs. (27.2±2.2) vs. (21.7±3.0)], and of group DM2 was significantly higher than that of DM1 group (P<0.01 all); expression of TNF-α in group DM2 and group DM1 were significantly higher than that of healthy control group [(27.5±1.5) vs. (20.2±2.7) vs. (13.1±1.6)], and of DM2 group was significantly higher than that of group DM1 (P<0.01 all)；expression of NF-κB in group DM2 and group DM1 were significantly higher than that of healthy control group [（30.1±1.7）vs.40.7±1.5）vs.（16.0±2.6）], but of group DM1 was significantly higher than that of group DM2 (P<0.01 all). Conclusion: There are obvious metabolic disorders of glucose and lipid in T2DM rats, and complicated morphological changes of myocardial tissues similar to myocardial lesions in DM humans; the expressions of SUMO1, NF-κB and TNF-α significantly increase, suggest SUMO1 takes part in inflammatory reaction mediated by NF-κB, TNF-α in myocardial lesion of rat with T2DM，and may inhibit NF-κB, possesses effect of protect myocardium.

Objective To construct a eukaryotic expression plasmid of pcDNA 3.1-Ag85A-CD226, and to use it as DNA vaccine then further evaluate its immunogenicity through oral administration in a mouse model.Methods The CD226-PCR2.1-ToPo plasmid was used as the template to clone CD 226 gene by PCR.The CD226 gene was then inserted into pcDNA 3.1-Ag85A plasmid to construct the recombinant plas-mid of pcDNA3.1-Ag85A-CD226.After identified by restriction enzyme analysis and sequencing , the re-combinant plasmid was transfected into HEK 293 cells by using lipofection .The expression of Ag85A-CD226 gene in HEK293 cells was detected by RT-PCR, Western blot and indirect immunofluorescence assay .The purified recombinant plasmid was used to prepare the Ag 85A-CD226 DNA vaccine by liposomal encapsula-tion.The vaccine was administered intragastrically to mice .The activities of NK cells , the cytokine levels in the supernatants of spleen cell cultures and the mRNA level of cytokines in the intestines were evaluated to analyze the immunogenicity of Ag85A-CD226 DNA vaccine.Results The Ag85A-CD226 DNA vaccine was prepared successfully .The expression of Ag85A-CD226 fusion protein was detected in HEK293 cells.The activities of NK cells from mice vaccinated with Ag 85A-CD226 DNA vaccine were higher than those from other control groups (P<0.01).The level of TNF-α, IFN-γand IL-2 in the supernatants of spleen cell cul-tures and in the intestines were significantly up-regulated in comparison with other control groups ( P <0.01).The level of IL-4 in the supernatants of spleen cell cultures was down-regulated in the experimental group (P<0.01), but the level of IL-4 in intestines showed no significant difference among the five groups (P>0.05).Conclusion The Ag85A-CD226 DNA vaccine could significantly enhance Th1 type immune responses systemically and in the intestine as in comparison with those vaccinated with single dose of Ag 85A DNA vaccine or CD226 DNA vaccine.

Objective: To study expressions of small ubiquitin-related modifier protein（SUMO）4 (SUMO4), nuclear factor (NF)- κB and inhibitory factor of NF-κB (IκB) in kidneys of rats with type 2 diabetes mellitus (T2DM). Methods: A total of ten 40-week-old male Goto-Kakizaki (GK) rats （with spontaneous diabetes mellitus）of specific-pathogen free (SPF) grade, and ten 40-week-old male Wistar rats of SPF grade were selected. The lesion of renal tissue was observed by hematoxylin eosin (HE) staining. Expressions of SUMO4, NF-κB and IκB in renal tissue were observed by immunohistochemistry methods. Results: In the GK rats, glomerular capillary ball hypertrophy, basilar membrane slightly thickening; glomerular mesangial cells hyperplasia, hypertrophy and renal tubular epithelial cells hypertrophy were observed. Compared with normal Wistar rats, expression levels of NF-κB [(0.232±0.034) vs. (0.634±0.058)], IκB [(0.242±0.027) vs. (0.712±0.078)] and SUMO4 [(0.160±0.031) vs. (0.545±0.045)] significantly increased in renal tissue of GK rats (P<0.01 all). Conclusion: Compared with Wistar rats, expressions of NF-κB, IκB and SUMO4 significantly increase in renal tissue of GK rats, suggesting that SUMO inhibiting transcriptional activity of NF-κB may exist in kidneys of T2DM rats. Therefore, sumoylation may be a new therapeutic target for inhibit renal microvascular lesion of diabetic disease.

Objective: To study influence of overweight and obesity on blood coagulation and metabolic disorders in patients with type 2 diabetes mellitus (T2DM). Methods: A total of 248 preliminary diagnosed T2DM patients were selected. According to body mass index (BMI), they were divided into normal BMI control group (n=95), overweight group (n=87) and obesity group (n=66). Blood lipids, blood glucose and fasting insulin (FINS) were measured in all patients and homeostasis model-insulin resistance index (HOMA-IR) was calculated then. Statistical analysis was performed. Results: Compared with normal BMI control group, there were significant increase in fibrinogen [(3.37±0.55) g/L vs. (4.04±0.70) g/L vs. (5.20±0.69) g/L], urine microalbumin [(14.46±8.90) mg/g vs. (47.33±42.54) mg/g vs. (104.45±60.78) mg/g], fasting blood glucose [ (7.15±0.97) mmol/L vs. (8.84±1.81) mmol/L vs. (10.06±2.28) mmol/L], FINS [(10.09±8.21) IU/ml vs. (14.33±15.55) IU/ml vs. (19.69±10.86) IU/ml], HOMA-IR[(3.19±2.59) vs. (5.51±5.38) vs. (8.48±4.62)], TG, TC and LDL-C levels in overweight group and obesity group, and the more BMI patients were, the higher these indicators were; There were significant decrease in plasma prothrombin time [(13.33±0.69)s vs. (12.74±0.69)s vs. (11.43±0.53)s], activated partial thromboplastin time [ (37.32±2.31)s vs. (36.55±2.41)s vs. (34.61±1.53)s] and HDL-C [(1.54±1.12) mmol/L vs. (1.27±0.41) mmol/L vs. (1.09±0.28) mmol/L] in overweight group and obesity group（P<0.05 all）. Conclusions: Overweight and obesity aggravate coagulation and metabolic disorders in patients with type 2 diabetes mellitus. It also aggravates degree of insulin resistance, the more BMI patients are, the more serious they are.

BACKGROUND:Fibronectin type Ⅲ domain-containing protein 5(FNDC5)is a muscle factor that can regulate glucose and lipid metabolism,and exert anti-inflammatory,anti-oxidative effects and improvement in insulin resistance.Moreover,FNDC5 could control various cell pyroptosis. OBJECTIVE:To explore the effect and potential mechanism of FNDC5 on macrophage pyroptosis. METHODS:(1)After completing the construction of the lentivirus virus overexpressing FNDC5 or silencing FNDC5,the THP-1 cells were transfected with the lentivirus vector.The result of transfection was detected by the expression of green fluorescence,qPCR,and western blot assay.(2)Phorbol ester induced THP-1 cells to differentiate into macrophages.Oxidized low-density lipoprotein(ox-LDL)was added to induce the cell pyroptosis model.There were six groups,i.e.,NC group,ox-LDL group,ox-LDL+MOCK1 group,ox-LDL+Ov-FNDC5 group,ox-LDL+MOCK2 group,and ox-LDL+shFNDC5 group.(3)The level of cell pyroptosis was evaluated by Hoechst 33342/propidium iodide fluorescence double staining and lactate dehydrogenase release assay.The expression levels of related molecules in THP-1 cells were analyzed by qPCR and western blot assay.The interleukin-18 and interleukin-1β in cell supernatant were detected by ELISA. RESULTS AND CONCLUSION:Compared with the ox-LDL+MOCK1 group,overexpression of FNDC5 significantly reduced the pyroptosis rate of macrophages and the release levels of lactate dehydrogenase,interleukin-1β and interleukin-18,significantly inhibited the mRNA expression of NF-κB p65,NF-κB p50,NLRP3,ASC,Caspase-1,and GSDMD,and significantly inhibited the protein expression of NF-κB p65,NF-κB p50,NLRP3,ASC,cleaved Caspase-1 and GSDMD-N.Compared with the ox-LDL+MOCK2 group,the silence of FNDC5 showed the opposite result.These findings suggest that FNDC5 attenuates pyroptosis in macrophages by inhibiting the NF-κB/NLRP3 pathway.