Objective To investigate the role of TRAF2 in AP-1 signaling pathway of human B cells.Methods Human Ramos B cells were transfected with plasmids expressing YFP fusion wild type TRAF2(YFP-WT-TRAF2) or YFP fusion dominant-negative TRAF2(DN-TRAF2),or transfected with shRNA-TRAF2 and control shRNA plasmid.After incubation overnight,cells were either sorted with flowcytometry or screened by antibiotics G418.Activation of AP-1 pathway,including phosphorylation of ERK,JNK and P38,as well as nuclear translocation of AP-1 subunits were detected by western blot and ELISA.Results Overexpression of WT-TRAF2 selectively induced activition of MAPK by phosphorylation of ERK and P38,and further induced nuclear translocation of C-FOS.Moreover,both overexpression of DN-TRAF2 and transfection of shRNA-TRAF2 inhibited phosphorylation of ERK and P38,and nuclear translocation of C-FOS.Conclusion TRAF2 selectively activates some kinases in CD40 mediated AP-1 signaling pathway,and plays an important role in AP-1 activation.

Objective To select the optimum bases for Sihuang Gao and study the method for quality control.Method Ultra-fine powders of Radix Scutellariae,Cortex Phellodendri,Radix et Rhizoma Rhei and Rhizoma Coptidis were used to prepare Sihuang Gao with different formulations,the bacteriostatic action of which were compared and stability were inspected.The best prescription was identified by TLC,and the skin irritation was observed.Result Sihuang Gao made of water-soluble base had a strong antibacterial activity and stable quality,and it was proved no irritation,the contents of active component could be detected by TLC.Conclusion The quality was controllable,and the improvement prescription can be used in the clinic.

Objective To optimize the preparation process of Xiaochuan Tiegao. Methods The effective compound imporatorin of Dahurian Angelica Root was determined by HPLC. The best condition of preparation process for the ethanol-extraction method was investigated by L9(34) orthogonal design. Results The optimum technical condition:adding 2 times ethanol, cooking 1 hour and 2 times with 90% ethanol. Conclusion The method is reliable, accurate and easy to operate.

Objective To develop an HPLC method for the assay of sinapine thiocyanate in Semen Sinapis Albae of different processed products. Methods It was assayed by RP-HPLC. The column was Hypersil BDS Phenyl (5 ?m, 250 mm?4.6 mm), the mobile phase was acetonitrile-0.08 mol/L KH2PO4 (15∶85). The UV detection wavelength was 240 nm. The flow rate was 1.0 mL/min. Results Good linearity of sinapine thiocyanate was showed within the range of 0.8～4.0 ?g (r=0.999 7). The average recovery was 100.98% (RSD=3.51%). Conclusion The method is simple, accurate and good reproducible. It can be used for quality control of Semen Sinapis Albae of different processed products.

Objective To investigate the clinical characteristics of gliomas in children,and assess the value of CT and MRI for diagnosing glioma.Methods Fifty-three children under 14 years old who were admitted into department of neurosurgery of Henan Provincial People's Hospital from Jan.2001 to Dec.2006 were diagnosed as glioma.Their age and sex distribution,clinical symptoms and signs,imaging examinations and pathological examinations were retrospectively analyzed.Results 1.There were 35 males and 18 females.2.The common symptoms and signs were headache and dizziness(32 cases,60.4%),nausea and vomiting(32 cases,60.4%),balance function disorder(31cases,58.5%),episodic convulsion(6 cases,11.3%) and fundus changes(33 cases,62.3%).3.There were 24 gliomas located at supratentorial and 29 gliomas located at infertentorial in this group.4.Thirty-three cases had a CT examination(1 case was wrongly diagnosed) and 28 cases had a MRI examination(all cases were positive);The positive rate of CT for displaying these tumors was 97%,MRI was 100%.5.Pathological examination revealed gliomas in 42 cases after operation.The most common histologically confirmed tumor type reported was astrocytoma,which accounted for 40.5%(17 cases),and the second was medulloblastoma,which accounted for 31.0%(13 cases).Conclusions 1.Glioma are more frequently affected in males than females in children.2.It often occurs in cranial posterior fossa and midline,which causes the symptoms of intracranial hypertension including headache,dizziness,nausea,vomiting,fundus changes,and so on.3.It often occurs in infertentorial,which causes the symptoms of balance function disorder,since it can invade cerebellum directly or indirectly.4.CT and MRI play an important role in the diagnosis of glioma in children.MRI is more valuable than CT in glioma diagnosis.5.Histological types of glioma in children are significantly different from adults.The most common histological types are astrocytoma and medulloblastoma.

Objective To investigate the effects of CD40-P227A SNP at human B cell signaling and function. Methods Human Ramos B cells were transfected with plasmids of CFP fusion wild type CD40 and mutant type CD40-P227A, together with plasmids of luciferase fusion NF-κB and AP-1, as well as plasmids of GFP fusion NF-κB. Activation of NF-κB and AP-1 pathway were detected by luciferase assay and flow cytometry, degradation of IκBα and nuclear translocation of NF-κB subunits were tested by Western blot and transfactor ELISA assay. Results Compared with wild type CD40, CD40-P227A SNP induced more degradation of IκBα, increased nuclear translocation of p65, p50 and c-Rel. As well as higher activity of NF-κB, as shown by increased NF-κB luciferase and GFP-NF-κB expressing B cells. Moreover, CD40-P227A SNP induced more activation of AP-1 pathway than CD40. CD40-P227A enhanced B cells function by inducing more excretion of IL-6 and TNF-α. Conclusion Our data indicate that CD40-P227A is a gain-of-function phenotype which induces activation of NF-κB and AP-1 signaling pathways and may contribute to the pathogenesis of the SLE autoimmune disease.

Objective To investigate the role of TRAF6 in NF-?B and AP-1 signaling pathway in human B cell line.Methods Human Ramos B cells were transfected with plasmids expressing YFP fusion dominant-negative TRAF6(DN-TRAF6),or transfected with shRNA-TRAF6 plasmid.After incubation overnight,cells were either sorted with flowcytometry or screened by G418.Activation of NF-?B and AP-1 pathway,including phosphorylation of I?B?,ERK,JNK and P38,as well as nuclear translocation of NF-?B subunits(P65,P50 and c-Rel)and AP-1 subunits(C-FOS,C-JUN,CREB and ATF) were detected by Western blot and ELISA.Results In cells which overexpress DN-TRAF6 or endogenous TRAF6 expression were knocked-down by shRNA,the phosphorylation of I?B?,as well as phosphorylation of JNK were inhibited.Furthermore,nuclear translocation of NF-?B subunits P65,P50,c-Rel,and AP-1 subunits C-FOS and C-JUN were also inhibited in B cells through overexpression of DN-TRAF6.Conclusion TRAF6 selectively activates some kinases in CD40 mediated NF-?B and AP-1 signaling pathway,and plays an important role in their activation.

Objective To compare the phenotypes of abnormal CD4+CD25-Foxp3+ T cells with traditional regulatory T cells (CD4+CD25+Foxp3+) in patients with untreated new-onset lupus (UNoL) and investigate their clinical relevance. Methods The expressions of surface markers (CD25, CD127, CCR4, GITR, CTLA-4) and intracellular marker(Foxp3) on the peripheral blood mononuclear cells from twenty-two UNoL patients were analyzed by flow cytometry analysis, and their clinical relevance were assessed. Results There were no significant differences between CD4+CD25-Foxp3+ and CD4+CD25+Foxp3- T cells in the expressions of GITR, CTLA--4 and CCR4 (P>0.05), but they were significantly lower than those of CD4+CD25+ Foxp3+ T cells in UNoL patients (P<0.01). The percentages of CD127low- in CD4+Foxp3+CD25high,CD4+Foxp3+ CD25low and CD4+Foxp3+CD25+ T cells were (93.8±3.5 )%, (93.7±2.3)% and (92.0±2.1)% respectively (P> 0.05), whereas the expressions of Foxp3 on CD4+CD127low- T subpopulations showed significant differences in CD4+CDI27low-CD25high (91.4±2.6)%, CD4+CD127low-CD25low (71.9±3.3)% and CD4+CD127low-CD25- (9.0± 2.2)% T cells(P<0.01 ). The frequency of CD+CCR4+CD25high T cells correlated negatively with SLEDAI (r=-0.695,P<0.001).and it was significantly lower in lupus nephritis patients(1.10±0.17)%compared with SLE patients without nephritis [(1.61±0.23)%,P<0.01]and healthy controls [(1.75±0.10)%,P<0.01], furthermore,the frequency of CD4+CCR4+CD25low-T cells in lupus nephritis was significantly higher than that in healthy controls[(11.5±2.3)%vs (8.0±1.0)%,P<0.01].Conclusion The increased CD4+CD25-Foxp3+ T cells in the Untreated Newonset Lupus(UNoL)patients mimic activated T effector cells.CD4+CD25high-CD127low-T cells can be used to isolate live CD4+CD25highFoxp3+regulatory T cells.CCR4+regulatory T cells may be involved in the pathogenesis of lupus nephritis.

Female ; Food Contamination ; Humans ; Infant ; Infant Formula ; Kidney Calculi ; chemically induced ; diagnosis ; Male ; Triazines ; toxicity

Objective To determine the expression level of CD154 in systemic lupus erythematosus(SLE) and its role in the pathogenesis. Methods CD154 were detected by flow cytometry in CD19+ peripheral blood lymphocytes from 16 active lupus patients and 14 healthy controls. Effects of anti-CD154, on the proliferation and IgG secretion by peripheral B lymphocytes were also observed. Results ① The positive rates and mean fluroscence intensity of CD154 in peripheral blood B lymphocytes from active lupus [(36?17)%,364?238] were significantly higher than from normal controls[(10?8)%, 124?97], (P