It has been shown that two pathways of signal transduction are activated under the stimulation of cytokines , such as IL-1?TNF-? ?EGF?TGF-? ?PDGF?IGF-1, secreted by fibroblast-like synoviocytes with rheumatoid arthritis. They are receptor tyrosine kinases-Ras-MAPK pathway and non- receptor tyrosine kinases-JAK-STAT pathway. The former activation plays an important role in chronic synovitis of rheumatoid arthritis. More researches are expected to focus on G-protein-AC-cAMP signal transduction pathway and the internal relations of all kinds of pathways.

10.3969/j.issn.2095-4344.2013.25.019

Aim To develop a convenient and effective method to isolate and culture primary rabbit aortic vascular smooth muscle cells(VSMCs).Methods The thoracic aortas were removed by dissection under sterile conditions.Aortic smooth muscle cells were excised and cleaned of fat and connective tissue,and the isolated vessel media was cut into 1 mm3 pieces.The explants were digested with different concentrations of collagenase typeⅠ,and incubated at 37℃ for different time,then undispersed explants were placed onto a sterile 100-mm plastic tissue culture dish with growth medium.Results VSMCs could emigrate from the explants digested 6 h by collagenase typeⅠ(1.5 g?L-1)for 24 hours,the cells would passage for another 4~5 days.Confluency could be reached within 3~4 days after subculturing.VSMCs were identified by immunoreactivity with ?-actin and by the smooth muscle cell-specific,hills and valley-like morphology.Conclusion It was an effective method to culture primary VSMCs from the explants digested for 6h by collagenase type Ⅰ(1.5 g?L-1),which could shorten primary culture time.

Aim To investigate the effects of paeonol on lipopolysaccharide ( LPS) and adenosine 5′-triphos-phate ( ATP) induced NLRP3 inflammasome activation in primary rat microglia and the mechanisms responsi-ble for this anti-inflammatory effects. Methods Pri-mary rat microglia were identified immunohistochemi-cally using the cluster of differentiation 11 b ( CD11 b ) antibody. Proinflammatory cytokine IL-1β was deter-mined by ELISA. Western blot was performed to ob-serve the protein expression of NLRP3 , ASC and caspase-1 in cultured primary rat microglia. The level of intracellular reactive oxygen species ( ROS) was mo-nitored by using the fluorescent probe 2′, 7′-dichlo-rofluorescein diacetate ( DCFH-DA ) . Results LPS (0. 5 mg · L-1 )/ATP ( 5 mmol · L-1 ) significantly increased intracellular ROS level and IL-1β secretion and upregulated NLRP3 , ASC and caspase-1 protein expression in primary rat microglia. Paeonol signifi-cantly decreased intracellular ROS level and IL-1β se-cretion, and inhibited LPS/ATP induced overexpres-sion of NLRP3 , ASC and caspase-1 in cultured primary rat microglia. Conclusion Paeonol can inhibit LPS/ATP induced NLRP3 inflammasome activation in pri-mary rat microglia, and this inhibitory effect may be through the suppression of intracellular ROS.

Aim To develop a convenient and effective method to isolate and culture primary rat aortic endothelial cells (RAECs). Methods The thoracic aortas were removed by dissection under sterile conditions. Aortas were turned over to expose the luminal surface, and the surfaces were digested with different concentrations of collagenase typeⅠ, incubated at 37℃ for different times, then, cut into pieces and placed luminal side down onto collagen-coated flask with growth medium. Results RAECs could emigrate from explants digested 1h by collagenase typeⅠ(2.0 g?L-1) for 24 h and cells would passage for another 4~5 days. Reached confluency within 3~4 d after subculturing. RAECs were identified by immunoreactivity with Factor-Ⅷ and by the endothelial cell-specific, cobblestone-like morphology. Conclusion It is an effective method to culture primary RAECs from explants digested for 1h by collagenase type Ⅰ(2.0 g?L-1),that can shorten primary culture time.

AIM To observe the antinociceptive effects of glucosides of Chaenomeles speciosa(GCS) and to study their relative mechanism. METHODS The effects of GCS on normal and inflammatory animals were observed in mouse acetic acid writhing test, mouse formalin test and arthritic flexion test of adjuvant arthritis(AA) rats; the concentration of prostaglandin E2 (PGE2) and tumor necrosis factor-α(TNF-α) secreted by the synovial cells of AA rats were measured by radioimmunoassay. RESULTSDifferent doses of GCS (60, 120, 240 mg·kg-1 for mice and 30, 60, 120 mg·kg-1 for rats, ig) inhibited mice′s writhing response and second phase of formalin response. It also suppressed the increased arthritic flexion scores in AA rats. On 28 d after inflammation induction, GCS (60, 120 mg·kg-1) decreased the concentration of PGE2 and TNF-α of synovial cells in the AA rats. CONCLUSIONGCS have antinociceptive effects, which related to its inhibitory effects on peripheral inflammatory mediators.

Aim To investigate the effects and mechanisms of glucosides of chaenomeles speciosa (GCS) in mice with contact hypersensitivity (CHS) response. Methods CHS model in mice induced by 2,4-dinitro-I-dinitroflurobenzene (DNFB) was used in this study. Concanavalin A (Con A)-induced splenocytes proliferation was measured by the MTT colorimetric method and interleukin-2 (IL-2) activity was measured by testing its ability to support ConA-induced mice splenocytes proliferation by MTT method. Interleukin-4 (IL-4) and transforming growth factor -beta 1 (TGF-? 1) levels were determined by ELISA. T lymphocytes subsets were measured by flow cytometry. Results Similar as the control drug 4-acetylaminophenylacetic acid (actarit or Acta) (120 mg?kg -1), GCS (240 mg?kg -1) could inhibit the thymus index and spleen index in CHS mice. GCS( 60,120,240 mg?kg -1) could inhibit the ear swelling of CHS mice and could inhibit the splenocytes proliferation induced by ConA. GCS (240 mg?kg -1)decreased CD4 +CD8 +T lymphocytes subsets ratio and resumed the CD4 +CD8 -subsets ratio in CHS mice;GCS(240,60 mg?kg -1) resumed the CD4 -CD8 -subsets ratio in CHS mice. GCS also decreased the TGF-? 1 and IL-2 levels and increased the IL-4 levels in mice thymus with CHS. Conclusion GCS could inhibit mice CHS reaction and resume the balance of T lymphocytes subsets in mice thymus with CHS, and also could modulate the cytokines production by CD4 + T lymphocytes of CHS mice.

AIM To investigate the therapeutic effect and mechanism of glucosides of Chaenomeles speciosa (GCS) on adjuvant arthritis (AA) in rats. METHODS Complete Freund's adjuvant was used to induce AA in rats. Secondary paw swelling of AA rats was measured with volume meter. Pain response and polyarthritis index were scored. Meanwhile, ConA induced thymocyte proliferation and LPS induced splenocyte proliferation were assayed by 3 (4,5 dimethylthiazol 2 thiazolyl) 2,5 diphenyl 2H tetrazolium bromide (MTT) assay. IL 1 production was measured by thymocyte proliferation assay; TNF ? and PGE 2 production were determined by radio immunoassay. RESULTS GCS (30, 60, 120 mg?kg -1 ) and Actarit (60 mg?kg -1 ) were given by intragastric administration for 8 days from the 17th day after immunization. Treatment with GCS (60, 120 mg?kg -1 ) and Actarit (60 mg?kg -1 ) for 5 days significantly diminished the secondary hind paw swelling, as well as relieved the pain response and the polyarthritic symptoms of the whole body as compared with AA model group. GCS (120 mg?kg -1 ) and Actarit (60 mg?kg -1 ) restored the diminished thymocytes proliferation ConA induced in AA rats. GCS reduced the enhanced productions of IL 1,TNF ? and PGE 2 from peritoneal macrophages in AA rats. CONCLUSION GCS has therapeutic effect on AA in rats, which may be related to regulation function of thymocyte T cells and inhibiting the productions of proimflammatory cytokines secreted by peritoneal macrophages.

Objective To evaluate the efficacy of itraconazole oral solution for prevention of invasive fungal infection ( IFI ) in neutropenic patients with acute leukemia after chemotherapy.Methods Clinical data of 136 neutropenic patients with acute leukemia after chemotherapy at the Department of Hematology,Nanfang Hospital from January 2008 to December 2010 were retrospectively analyzed.Patients were divided into itraconazole group ( n =67 ) and control group ( n =69).There were 36 patients with acute nonlymphocytic leukemia ( ANLL),31 with acute lymphoblastic leukemia (ALL) in itraconazole group;while in control group,there were 30 patients with ANLL,38 with ALL and 1 with biphenotypic acute leukaemia (BAL).Patients in itraconazole group received intraconazole after chemotherapy until the neutrophil count was increased to 0.5 × 109/L or the body temperature returned to normal and without any imaging evidence of IFI.The incidence of IFI and clinical features were compared between the groups using SPSS 13.0 software.Pearson x2 test was used for nominal variables,for measurement data,t (normal distribution) or Mann-Whitney U (skewed distribution) test were used.Results There were 12 cases ( 17.9％ ) suffering from IFI in itraconazole group and 32 cases (46.4％) in the control group (x2 =12.59,P ＜ 0.01 ).For ANLL patients,the incidence of IFI in itraconazole group was significantly lower than that in control group ( 16.7％ vs.56.7％,x2 =11.53,P ＜0.01 ).In itraconazole group,the incidence of IFI in female patients was significantly lower than that in male patients ( 8.6％ vs.28.1％,x2 =4.35,P ＜0.05 ).And for the female patients,the incidence of IFI in itraconazole group was significantly lower than thatin the control group (8.6％ vs.44.7％,x2 =11.98,P＜0.01).Conclusion Itranconzole oral solution can effectively prevent IFI in neutropenic patients with acute leukemia after chemotherapy,especially for the female patients with ANLL.

Objective To investigate the effects of high flow hemodialysis (HFHD) on cardiac function in uremia patients. Methods A prospective randomized controlled study was conducted. Sixty patients who were diagnosed with uremia, taken maintenance hemodialysis (MHD) and 30 healthy controls admitted to the Second People's Hospital of Guiyang from December 2014 to June 2015 were enrolled. They were randomly divided into two groups:HFHD group (HFHD three times a week) and the routine hemodialysis group (HD group, HD three times a week), with 30 in each group. Patients in each group were received hemoperfusion and hemofiltration once a month. Before the treatment and 6 months after the treatment, venous blood from all the patients were collected for testing the brain natriuretic peptide (BNP), cardiac troponin T (cTnT) and the ultrasound cardiograph were done at the same period by a special person, the left ventricular end diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD), the left ventricular end diastolic volume (LVEDV), left ventricular end systolic volume (LVESV), left ventricular posterior wall thickness (LVPWT), interventricular septum thickness (IVST), early and late diastolic blood flow to the largest ratio (E/A), left ventricular ejection fraction (LVEF), left ventricular mass index (LVMI) were recorded. Results Compared with the health control group, BNP, cTnT, LVEDD, LVESD, LVESV, LVPWT, IVST were significantly increased, LVEDV were significantly lowered before treatment in the HD group and HFHD group. But no significant differences in the above indexes and E/A, LVEF, LVMI between two groups were found. Compared with the data before treatment, the BNP, LVPWT were significantly lowered after treatment in HD group [BNP (ng/L): 641.50±60.09 vs. 2676.20±454.30, LVPWT (mm): 10.57±1.16 vs. 12.57±1.41, both P < 0.05]. The BNP, LVPWT were significantly lowered in HFHD group as compared with HD group [BNP (ng/L): 253.10±48.77 vs. 641.50±60.09, LVPWT (mm): 9.29±1.08 vs. 10.57±1.16, both P < 0.05]; in addition, the cTnT, IVST, LVMI were significantly lowered after the treatment in HFHD group compared with those before treatment [cTnT (μg/L): 0.014±0.005 vs. 0.028±0.011, IVST (mm): 7.81±1.69 vs. 11.04±2.23, LVMI (g/m2): 149.10±15.77 vs. 158.70±17.25, all P < 0.05], and the LVEF were significantly increased in HFHD group as compared with those before treatment (0.574±0.068 vs. 0.528±0.082, P < 0.05). Conclusion HFHD has obvious advantages than the routine HD in improving cardiac function of uremia patients.