Objective To discuss the role of p38MAPK signal pathway in the process of TWEAK inducing rheumatoid arthritis(RA) fibroblast-like synoviocyte(FLS) to synthesize MMP-9 and look for a new target for RA treatment. Methods RA FLS were primarily cultured and stimulated with TWEAK. Western blot was used to detect the expression of p-p38MAPK and p65 in RA FLS. FLS were pretreated by SB203580 or not. ELISA was used to detect the concentration of MMP-9 in cell-cultured fluid. The mRNA expression of MMP-9 was measured by RT-PCR. Results TWEAK( 100 ng/ml) can make p38MAPK phosphorylated and increased the expression of p65 protein in the cell nucleus. SB203580 can partially inhibit the expression of MMP-9 and MMP-9 mRNA produced by RA FLS which is induced by TWEAK. Conclusion TWEAK induced RA FLS to synthesize MMP-9, in that process, the p38MAPK signal trausduction pathway was in active state, and induced the expression of NF-κB.

Objective: To prepare and optimize the prescription of betulinic acid (BA) ethosomes modified by sodium deoxycholate (SDC) and then to investigate its transdermal penetration as carrier of BA. Methods: The BA ethosomes modified by SDC were prepared by the ethanol injection method. The encapsulation efficiency (EE) was considered as the evaluation index to optimize the prescription of the ethosomes by orthogonal design, and the shape and particle size of the optimized ethosomes were analyzed. The in vitro transdermal absorption of BA was evaluated using Franz diffusion cells. The accumulated permeation amounts and permeation rate of liposomes, ethosomes, and BA ethosomes modified by SDC were compared. Results: The best formulation consisted of soybean lecithin-SDC-BC (18:1:1) and 35% ethanol. The average EE and the particle size were (93.8 ± 1.6)% and (102.3 ± 3.6) nm, respectively. The accumulated permeation amount of BA ethosomes modified by SDC in 12 h was (99.62 ± 9.44) μg/cm2, which was 1.67, 3.85, and 8.33 times of ethosomes, liposomes, and satuated solution containing 10% isopropanol, respectively. Conclusion: The BA ethosomes, modified by SDC with high EE, obviously enhance the percutaneous absorption of BA and might be one of the most perspective percutaneous preparations.

Objective To study whether p38 mitogen-activated protein kinases (p38MAPK) signal pathway were activated in the process of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) which then induces rheumatoid arthritis (RA) fibroblast-like synoviocyte (FLS) to synthesize matrix metalloproteinase-1 (MMP-1) and look for the relative mechanisms of how TWEAK was involved in the destruction of articular bones and cartilage. Methods RA FLS were primarily cultured and stimulated with TWEAK. FLS were pretreated with SB203580 or not. ELISA was used to detect the concentration of MMP-1 in cell-cultured supernatant. Western blotting was used to detect the expression of p-p38MAPK and P65 in RA FLS. Results TWEAK (100 ng/ml) could induce RA FIS to synthesize MMP-1. SB203580 could partially inhibite the expression of MMP-1 producted by RA FLS which was induced by TWEAK. TWEAK could make p38MAPK phosphorylated and increase the expression of P65 protein in the cell nucleus. Conclusion TWEAK induces RA FLS to synthesize MMP-1. In this process, p38MAPK signal transduction pathway is activated and then induce the expression of NF-κB.

Female ; Granulomatosis with Polyangiitis ; complications ; diagnosis ; Humans ; Otitis Media ; diagnosis ; etiology ; Young Adult

Objective To explore the application of nurse-administered psychological assessment scale for patients early after trauma in the wounded after Wenchuan earthquake.Methods The clinically commonly used self-rating scales (integrated SCL-90,SDS and SAS,and mental status scale in non-psychiatric settings),clinician-administered scale (nurse-administered psychological assessment scale for patients early after trauma)and diagnostic criteria in psychiatric settings(CCMD-3)were employed to assess the mental status of 21 patients after Wenchuan earthquake,and the recovery rates of these scales were analysed.Results All the 21 patients suffered from psychological disturbance to some extent, while none met the criteria of mental diseases.The recovery rate of the integrated SCL-90,SDS and SAS was only 9.5%, while those of the nurse-administered psychological assessment scale for patients early after trauma and mental status scale in non-psychiatric settings reached 100%.Conclusion The nurse-administered psychological assessment scale for patients early after trauma and mental status scale in non-psychiatric settings with fewer items are more suitable for the wounded early after earthquake,with a favourable confidence and efficacy.The diagnostic criteria in psychiatric settings can not be universally used in the wounded after earthquake.

Background Research showed that exposure of 530 nm monochromatic light can induce myopia in animal,and retinal Müller cells participate in the formation of myopia.However,the effect and mechanism of retinal Müller cells during the formation of monochromatic light induced-myopia is below understood.Objective This study was to investigate biologic characteristics of rat retina Müller cells and the expression of cell factors in Müller cells after being illuminated by the 530 nm monochromatic light,and discuss the role of the retina Müller cells in myopia induced by monochromatic light.Methods Immortalized rat retinal Müller cells were cultured with DMEM containing 10％ fetal bovine serum in a self-made cell incubator with monochromatic light by adjusting luminance of 530 nm LED source.The cells were exposed to 125,250 and 500 lx luminance respectively for 6,12 and 24 hours,and the cells without light-irradiation were used as control.The growth of the cells under the different light time and different illuminations was described by MTT as the absorbance at the wavelength 570 nm (A570),and cell cycle analysis of Müller cells was performed by flow cytometry 48 hours after cultured,and the expression of transforming growth factor-beta 1 (TGF-β1),tyrosine hydroxylase(TH),inducible nitric oxide synthase(iNOS) and basic fibroblast growth factor(bFGF)in the cells were detected by reverse transcription PCR(RT-PCR),respectively.Results The Müller cells were uniform in size with polygonal shape and defined edges.No statistically significant difference was found in the A570 value in the cells of the 125 lx and 250 lx illuminated groups compared with the control group in various time points(P＞0.05).However,significant lowing was seen in the A570 value in the cells of the 500 lx illuminating for 12 hours and 24 hours in comparison with the control group (P =0.013,0.001).Compared with the control group,the ratio of the number between G2 and G1 phase was not significantly declined in 125 lx,250 lx illuminating for 48 hours (P =0.073,0.330),and the ratio in the 500 lx illuminating group was significantly lower than those in the 250 lx illuminated group and the control group (P =0.028,0.038).RT-PCR revealed that the expression of TGF-β1 mRNA in the cells was higher in the 250 lx illuminated group than that of the 500 lx illuminated group (P=0.006).The expression of iNOS mRNA was gradually upregulated in the 250 lx illuminated group compared with the control group (P =0.001),but that in the 500 lx illuminated group was downregulated (P =0.000).The expression of bFGF mRNA was raised in the 125 lx and 250 lx groups but reduced in the 500 lx group when compared with the control group(P=0.002,0.000,0.005).Also,the expression of TH mRNA was significantly increased in the 250 lx group(P=0.000),but decreased in the 500 lx group(P=0.000,P=0.001).Conclusions The monochromatic light of 530 nm can inhibit the growth of rat Müller cells and downregulate the expression of myopia-related cell factors and therefore exert effect in the formation of myopia.

Background The human transforming growth factor beta-induced gene (TGFBI) is the first determined pathogenic gene to corneal dystrophy.But the molecular genetic mechanism is completely unknown.The study of concerning role of TGFBI is very important for us understand the physiological function of cornea,and the pathogenesis of corneal dystrophy.Objective The vector of human transforming growth factor beta-induced gene (TGFBI) in eukaryotic expression was constructed and transfected into the human corneal epithelial cells in order to explore its influence on the growth of human corneal epithelial cells.Methods Total RNA was extracted from normal donor cornea tissue and cDNA was obtained by reverse transcription.TGFBI cDNA was synthesized by reverse transcription-PCR and cloned into pCMV-N-HA vector and identified by sequencing with PCR and EcoRV,XhoI double restriction endonuclease.The cells were grouped into recombinant pCMV-N-HA-TGFBI plasmid group,pCMVN-HA plasmid group,non-transfected group and pGFP-C2 transfected group.The recombinant pCMV-N-HA-TGFBI plasmid was transfected to human corneal epithelial cells and identified by observing the expression of enhanced green fluorescence protein(EGFP) in the cells.The TGFBI mRNA and proteins were harvested from the cells for real-time PCR analysis and Western blot assay respectively in 58 hours after transfection.The growth of the transfected cells was assessed by Cell Counting Kit-8.The expressions of matrix metalloproteinase(MMP) and tissue inhibitors of matrix metalloproteinase (TIMP) proteins and their mRNA in transfected cells were detected using SYBR fluorescence realtime PCR analysis and Western blot assay.Results The sequencing result of pCMV-N-HA-TGFBI positive clone plasmid showed that amplified TGFBI eDNA inserted into the vector at the correct sequence.EGFP was expressed in transfected cells in 48 hours after transfer of pGFP-C2 with the transfer efficacy 70％.The expression intensity of TGFBI mRNA was significantly higher in recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group,and TGFBI protein was expressed in recombinant pCMV-N-HA-TGFBI plasmid group.No significant difference was found in the A450value among recombinant pCMV-N-HA-TGFBI plasmid group,pCMV-N-HA plasmid group and non-transfected group ( F=3.34,P＞0.05 ).The mRNA level of MMP1,MMP3in the transfected cells was significant elevated but that of TIMP1 was declined in the recombinant pCMV-N-HA-TGFBI plasmid group compared with pCMV-N-HA plasmid group and non-transfected group (all P ＜ 0.05 ).Meanwhile,the expressions of MMP1,MMP3 and TIMP1 proteins appeared the same tendency( all P＜0.05).Conclusions Eukaryotic expression vector harboring human TGFBI eDNA can be successfully constructed and efficiently overexpressed in human corneal epithelial cells.TGFBI gene is involved in the physical and pathological conditions of human corneal epithelial cells by regulating the activity of MMP1,MMP3 and TIMP1.The results offer a new approach for the study of the role of TGFBI in pathogenesis of corneal transparency.

Background Wearing contaclenincreasethe risk of infection of the cornea.Some studieshowed the gas-permeability of materialused foconstructing corneal contaclenione of the contributing factorrelated to corneal health.Objective Thistudy wato observe the in vitro adherence ability of differenbacterito rigid gas-permeable contaclense(RGP-CL) made with varioumaterials.MethodContaclensemade with hexafocon,enflufocon opolymethyl methacrylate (PMMA) were placed into Staphylococcuaureus,Staphylococcuepidermidis,oPseudomonaaeruginosbacterial suspension(0.5 MCF) fo24 hours.The strength of bacterial adherence watested and studied by the methyl thiazolyl tetrazolium (MTT) colorimetrimethod based on absorbance (value),and the vortex method waused to calculate the colony forming units.The bactericlump formation waexamined with scanning electron microscope (SEM).ResultMTcolorimetrimethod showed thathe adherence ability of Staphylococcuaureuto hexafocon (value) wasignificantly lowethan thato enflufocon and PMMA,respectively (q=7.379,8.207,P＜0.01),buno significandifference wafound in the adherence ability of Staphylococcuaureubetween enflufocon and PMM(q =0.828,P＞0.05).The adherence ability of Staphylococcuepidermidito XO and enflufocon walowethan thato PMM(q =14.000,12.800,P＜0.01),buno significandifference wafound between the adherence of Staphylococcuepidermidito hexafocon and enflufocon material (q =1.200,P＞0.05).There wano significandifference in the adherence ability of Pseudomonaaeruginosto all three material(F=2.155,P=0.138).The vortex method presented the colony forming unitof Staphylococcuaureuto hexafocon,enflufocon and PMMwith (37.9± 1.5)×106,(49.9±2.2)×106 and (67.4± 1.6)×106,respectively,with significandifference among them (F =206.240,P＜0.01),showing the lowesvalue in hexafocon,the highesvalue in PMMand middle value in enflufocon (q=11.650,28.640,16.990,P＜0.01),Moreover,colony forming uniof Staphylococcuepidermidito hexafocon,enflufocon and PMMwa(7.9 ± 1.3) × 106,(10.5 ± 1.5) × 106,(11.2 ±1.2) × 106,respectively.And thaof hexafocon walowethan one of the PMMmaterial (q =5.060,P＜0.05).No significandifference wafound between hexafocon and enflufocon nobetween hexafocon and PMM(q =3.290,1.770,P＞0.05).In addition,the resultthacorresponded to the vortex method were seen in the MTcolorimetriassay (F =0.232,P =0.799).SEM examination showed dispersed population of Staphylococcuaureuand Staphylococcuepidermidion the surfaceof hexafocon and enflufocon;while much more Staphylococcuaureuand Staphylococcuepidermidiadhered on the surface of PMMA,forming net-like appearance.Conversely,high numbeof Pseudomonaaeruginoswaseen on the surface of all three materials,withounoticeable differencein the bacterial shape and quantity on each of the material.ConclusionThe adherence ability of bacterito PMMistrongethan thaof hexafocon and enflufocon,and gas-permeable material of RGP-CL doenoimpacthe adherence ability of bacteria.

OBJECTIVETo study the proliferation and apoptosis of tetramethylpyrazine (TMP) on leukemic U937 cells and its possible mechanism.

METHODThe inhibitory effect of TMP on the proliferation of U937 cells was detected by CCK-8 assay. The cell apoptosis and cycle distribution were examined by the flow cytometry. The mRNA expressions of bcl-2 and P27 were determined by the Real-time PCR. Western blot was carried out to detect bcl-2, caspase-3, cyclin E1, CDK2 and P27 expressions.

RESULTTMP inhibited the proliferation of U937 cells in a dose-and-time dependent manner, with IC50 value of 160 mg x L(-1) at 48 h. In addition, TMP could induce the apoptosis of U937 cells and block the cell cycle in G0/G1 phase. According to the results of Real-time PCR and Western blot, TMP could down-regulate the expression of apoptosis-related molecule bcl-2, cycle-related protein cyclin E1 and CDK2 and up-regulate caspase-3 and P27.

CONCLUSIONTMP shows the effects in inhibiting the proliferation of leukemic U937 cells and inducing the apoptosis. Its mechanism may be related to the impacts on the cell cycle distribution, down-regulation of the bcl-2 expression, which finally activates caspase-3, starts the apoptosis path and causes the cell apoptosis.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase 2 ; analysis ; Humans ; Leukemia ; drug therapy ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Pyrazines ; pharmacology ; therapeutic use ; U937 Cells

Objective To explore the association between inflammatory cytokines and islet β-cell function in chronic persistent asthma patients. Methods 112 adults with persistent asthma and 60 healthy volunteers were enrolled in this study. According to the severity of disease, all subjects were divided into persistent-mild group and persistent-moderate group. Plasma tumor necrosis factor-α(TNF-α), interleukin6(IL-6) and C-reactive protein (CRP) were determined. Oral glucose tolerance and insulin releasing test were performed. The ratio of the area under the curve of insulin to area under the curve of glucose ( AUC1/AUCG ), homeostasis model assessment for insulin resistance ( HOMA-IR), insulin sensitivity index( ISI),homeostasis model assessment for β-cell function (HBCI) and early insulin secretion index(△I30/△G30)were calculated. The values of forced expiratory volume in l second ( FEV1 ), forced vital capacity(FVC)and FEV1/FVC were recorded. Results In patient groups, the values for plasma TNF-α, IL-6, CRP,AUC1, AUC1/AUCG, HOMA-IR, HBCI significantly increased compared with those in control group, while ISI declined ( t =2. 02～13.62, P ＜0. 05). Multiple step regression analysis showed that HOMA-IR was positively correlated with CRP, LDL-C, BMI, AUC1, TNF-α( P ＜0. 01 orP ＜0. 05), but negatively correlated with FEV1 ( P ＜ 0. 05 ). Conclusion The results indicated that inflammatory cytokines ( TNF-α,IL-6,CRP) might result in insulin resistance in asthma patients who had hyperinsulinism at the same time.