Inflammation and bone erosion are suppressed in models of rheumatoid arthritis following treatment with a novel Syk inhibitor
Polly R.Pine a ,Betty Chang a ,Nathan Schoettler b ,Mona L.Banquerigo b ,Su Wang a ,Angela Lau a ,Feifei Zhao a ,Elliott B.Grossbard a ,Donald G.Payan a ,Ernest Brahn b,?
a
Rigel Pharmaceuticals,Inc.,1180Veterans Blvd.,San Francisco,CA 94080,USA
b UCLA School of Medicine (NS,MB,EB),Division of Rheumatology,1000Veteran Ave.Rm 32-59,Los Angeles,CA 90095-1670,USA
Received 6February 2007;accepted with revision 9March 2007Available online 29May 2007
Spleen tyrosine kinase (Syk)is an intracellular protein tyrosine kinase that serves as a key mediator of Fc receptor and B cell receptor signaling in inflammatory cells,including mast cells,macrophages,dendritic cells (DC),NK cells,and neutrophils.Through its interaction with the phosphorylated gamma chains of the heterodimeric Fc γreceptors,Syk is essential for the signal transduction initiated by activated receptors for immunoglobulin G.Aggregation of the Fc
Abbreviations:Syk,spleen tyrosine kinase;FcR,Fc receptor;RPA,reverse passive Arthus;RA,rheumatoid arthritis;IFA,incom-plete Freund's adjuvant;CIA,collagen-induced arthritis;COMP ,cartilage oligomeric matrix protein;ELISA,enzyme-linked immuno-sorbant assay;IgG,immunoglobulin;Micro-CT ,micro-computed tomography;LC,liquid chromatography;MS/MS,tandem mass spectrometry;TNF α,tumor necrosis alpha;MCP-1,monocyte chemotactic protein 1;Gro/KC,keratinocyte derived chemokine;BCR,B cell receptor .
?Corresponding author .Fax:+13102068606.
E-mail address:EBrahn@https://www.doczj.com/doc/9f17647363.html, (E.Brahn).
1521-6616/$–see front matter ?2007Elsevier Inc.All rights reserved.doi:10.1016/j.clim.2007.03.543
ava i l a b l e a t w w w.s c i e n c e d i r e c t.c o m
w w w.e l s ev i e r.c o m /l o c a t e /yc l i m
Clinical Immunology (2007)124,244–257
receptors,induced by antibody–antigen complexes,can induce many cellular functions including phagocytosis, degranulation,NK cell antibody-dependent cellular cyto-toxicity,and cytokine secretion,depending on the cell type. The initiation of inflammatory and destructive responses by these effector cells is believed to be crucial for the development and pathogenesis of autoimmunity[1].Immune complex(IC)-mediated acute inflammatory tissue injury has been implicated not only in rheumatoid arthritis(RA),but in a variety of human autoimmune diseases,including systemic lupus erythematosus and certain types of glomerulonephritis [2,3].
Several recent studies in Fc receptor(FcR)-deficient mouse strains suggest that signaling through this receptor is the critical step in the progression of tissue destruction in murine models of inflammation[4–7].In KRN TCR transgenic mice,in which the arthritis is chronic and symmetric with pannus formation and destructive lesions of joints and bones, FcγRIII-deficient mice have markedly diminished paw swel-ling following treatment with K/BxN serum,and mast cell-deficient mice have markedly attenuated synovitis[5].Ji et al.[8]confirmed that the arthritogenic immunoglobulins act through Fc receptors(FcγRIII and complement).Further-more,in mice immunized with collagen,on an arthritis susceptible DBA background,mice lacking FcRγchains were protected relative to wild-type controls,although both groups produced similar levels of anti-collagen antibody[6].
Despite extensive efforts,the etiology and pathogenesis of RA remain poorly understood,and numerous cell popula-tions have been implicated including T cells,B cells, monocytes/macrophages,mast cells,dendritic cells and fibroblasts[9].Chemokines,matrix metalloproteinases, adhesion molecules,angiogenic growth factors,and dysre-gulated intra-articular expression of proinflammatory cyto-kines(in particular IL-1βand TNFα)play key roles in the pathogenesis of RA[10].It is known that complement is activated in RA synovial fluids,and complement components are even locally produced[11].Moreover,immune complexes are deposited at the cartilage level[12].Humoral mediators may play a role as well;rheumatoid factors,found in90%of patients,are associated with a poor prognosis.Genetic linkage studies implicate FcR polymorphisms as a potentially important disease susceptibility factor.In humans,there are three classes of FcγR(I–III);the level of expression and localization of FcγR(activating and inhibitory)restricts specific biological properties to certain cell types.Functional polymorphic variants within FCRG2A or FCRG2B,or within FCGR3A or FCGR3B,may additionally contribute to disease susceptibility and pathogenesis[13–17].FcγR polymorphisms appear to be important predisposing factors in systemic lupus erythematosus(SLE)and multiple sclerosis(MS)[15,18].In addition,IgG receptor polymorphisms are also reported to influence the efficacy of intravenous immunoglobulin ther-apy for patients with autoimmune diseases[19].
The effects of a novel small molecule,R406,have been examined in several cell systems,and its activity is consistent with Syk inhibition in isolated mast cells, macrophages,DC,and synoviocytes.R406potently inhibits IgG and IgE-induced mast cell degranulation in cell-based assays with an EC50of approximately50nM[20,21]. Selectivity towards Syk inhibition was demonstrated in mast cells activated by FcεR1cross-linking,in which R406inhibited the phosphorylation of linker for activation of Tcell (LAT)tyrosine Y191(a Syk substrate)about50-fold more potently than the phosphorylation of Syk itself,which is phosphorylated by Lyn kinase.Subsequent signaling events in that pathway,including JNK and ERK1/2,but not p38,were also inhibited[21].IC stimulation of DC cytokine secretion is also inhibited by R406.R406prevented cytokine release(IL-13)from DC isolated from the bone marrow of ovalbumin (OVA)-sensitized mice and cultured in the presence of R406 and OVA-IgG immune complexes[22].Recent data implicate Syk signaling in TNFα-induced inflammatory cytokine and matrix metalloproteinase(MMP)production by fibroblast-like synoviocytes(FLS)isolated from joint tissues of RA patients[23],and treatment of cultured FLS with R406 markedly suppressed TNFα-induced IL-6and MMP-3produc-tion in RA FLS.Syk is an attractive target for immune and inflammatory diseases because inhibition of Syk can suppress numerous overlapping pathways simultaneously[1,23,24].
Given that Syk has a pivotal role in mediating FcR activation in hematopoietic cells,we investigated R406and R788(prodrug of R406)for their capacity to suppress the reverse passive Arthus(RPA)reaction and collagen-induced arthritis(CIA).
Materials and methods
Animals
Female Sprague–Dawley rats(Hilltop Laboratories,Scott-dale,PA),aged8–10weeks,were used in the Arthus studies. For CIA,female Lewis rats,aged6–8weeks,were purchased from Harlan Laboratories(Indianapolis,IN),and syngeneic LOUVAIN(LOU)rats were inbred at UCLA(Los Angeles,CA). In-life procedures were approved for each model by the respective Institutional Animal Care and Use Committee.
R788/R406
R406was synthesized and was also prepared as a soluble prodrug(R788).R406is soluble in a nonaqueous medium and was formulated(Tocopheryl Polyethylene Glycol1000 Succinate,TPGS:Propylene Glycol,PG,40:60,vol:vol)for in vivo administration;R788was prepared as a suspension in 0.1%carboxymethylcellulose,0.1%methylparaben–0.02% propylparaben–99.78%water.R788was found to be superior to R406in pharmaceutic properties critical for a drug product,exhibiting improved solubility and bioavailability. Following oral administration of R788,the active moiety is R406,and very low levels of the prodrug reach the systemic circulation.R788was prepared as the R406equivalent by weight.
Immune complex induction of the reverse passive Arthus(RPA)reaction,treatment with R788/R406, and measurement of inflammation and edema
Test article was administered to animals60min prior to an intravenous challenge with1%ovalbumin(OVA)in saline (10mg/kg)containing1%Evans blue dye.Ten minutes later, rabbit anti-OVA IgG(50μg/25μl)was injected intradermally
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(ID).Reagents were obtained from Sigma Chemical Co.(St. Louis,MO).Four hours after challenge,local skin edema was measured by dye extravasation into the surrounding tissue from an8-mm punch biopsy(OD610following incubation of tissue overnight in formamide).Edema was determined by the difference in each tissue biopsy injected with specific anti-OVA compared with tissue from a biopsy injected with non-specific IgG.
Induction of CIA
Lewis rats
CIA was induced in6-to7-week-old female Lewis rats.ID injection(10–15sites)of500μg bovine collagen II(CII, Chondrex,Inc.,Redmond,WA)emulsified1:1(vol:vol)with incomplete Freund adjuvant(IFA,Chondrex,Inc.)was used to initiate clinical arthritis in Lewis rats.On day7,immunized Lewis rats were boosted with CII/IFA(100μl emulsion administered ID).Arthritis normally appeared on one or both hind paw ankles in most rats between days10and12. Naive rats(n=4)were not immunized and did not receive drug treatment.
LOU rats
Syngeneic LOU rats,6–8weeks of age were maintained in the Department of Laboratory Animal Medicine at UCLA[25].ID injection(15–20sites)of500μg chicken CII in IFA(1:1)was used to immunize the LOU rats.LOU rats received a single immunization,and arthritis occurred in virtually all rats on day10.
Treatment with R788/R406
R406,R788,or vehicle was administered orally for18days to arthritic rats(b.i.d.for Lewis rats or qd for LOU rats). Pharmacokinetic parameters dictated an interval of at least 8h between the b.i.d.drug administration in the Lewis rats. The treatment started after disease onset within6h after individual rats showed a clinical score of“1”or higher.The enrollment day of each rat was designated as day0.For brevity,results of treatment with R406in Lewis rats and R788treatment in LOU rats are presented here;however, both agents were tested in each strain with similar results. R406and R788plasma levels were measured following extraction of protein using a mixture consisting of acetoni-trile,ethanol and dimethyl sulfoxide(DMSO)(2:1:1vol:vol). The extract was analyzed by LC-MS/MS.The C max was observed at1h for the low dose.The limit of quantitation (LOQ)was1.0ng/ml.
Assessment of clinical arthritis in CIA
The clinical arthritis was observed daily and the severity assessed by a semi-qualitative clinical score[25]as follows: 0,normal,without any macroscopic signs of arthritis;1, mild,but definite redness and swelling of the ankle,or apparent redness and swelling limited to individual digits, regardless of the number of affected digits;2,moderate redness and swelling of the ankle;3,redness and swelling of the entire paw including digits;4,maximally inflamed limb with involvement of multiple joints.In these studies,the maximum score was8,which was the sum of scores from both hind paws of each animal.
Antibodies to type II collagen by enzyme-linked immunosorbant assay(ELISA)
Antibody titers to type II collagen were assayed by ELISA using peroxidase-conjugated goat anti-rat IgG antibodies according to the manufacturer's instructions(Accurate Chemical and Scientific Co.,Westbury,NY).Alternatively, antibody titers to type II collagen were assayed by ELISA using a kit from Chondrex,Inc.(Redmond,WA).The limit of detection was b3ng/ml.
Antibodies to mouse anti-human TNFα
Antibody levels of anti-human TNFαIgG administered intravenously were assayed by ELISA using purified mouse monoclonal IgG(R&D Systems,Minneapolis,MN).Plates were coated with human TNFαat a concentration of1.0μg/ml, incubated with test serum,peroxidase-conjugated goat anti-mouse IgG-HRP antibody,and color developed with tetra-methylbenzidine(TMB).The reaction was stopped with50μl of1N sulfuric acid and the plates were read at450nm. Radiographic assessments
Radiographic severity of CIA was assessed blindly on day 28.High-resolution digital radiographs(48kV,2mA s)of hind limbs were performed on all animals.Rats were given a score of0–3for each hind limb with a summated maximum score of6based on the extent of soft tissue swelling,joint space narrowing,bone destruction,and periosteal new bone formation(0,normal;1+,soft tissue swelling only;2+soft tissue swelling and early erosions;3+, severe erosions).
Micro-CT analysis was performed on a representative subset of hind limbs excised from arthritic LOU rats treated with vehicle alone or30mg/kg/day R788for18days.The acquisition parameters were:70kVp,25mA,20-ms exposure time,500 views over200°.The three-dimensional image matrix was 1740×1740×910.After each rat paw was segmented from the primary micro-CT data,a three-dimensional volume rendering of the skeletal structures was created[26].
Cartilage oligomeric matrix protein(COMP)analysis
Serum COMP was measured using a competitive ELISA kit (Kamiya Biomedical Company,Seattle,WA)following proto-cols from the https://www.doczj.com/doc/9f17647363.html,P concentration was determined by using SoftMax Pro from Molecular Devices (Sunnyvale,CA).The limit of detection was b0.2U/l. Histologic analysis
Rat paws were fixed in phosphate-buffered10%formalde-hyde,decalcified,and one-half of each limb was processed in paraffin blocks and sectioned at approximately5μm.The tarsus and digits were cut in a sagittal plane.All sections were stained with Safranin O and examined by light microscopy.
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Scoring(0–17)was performed by a single blinded observer using a modified Mankin score[27],as outlined in Fig.7F. Immunohistochemistry and Western blot analysis
of Syk
Deparaffinized sections(tarsus)were stained with H&E, blocked with1.5%normal goat serum(Vector Labs,Burlin-game,CA),incubated with polyclonal antibody to Syk(Cell Signaling,Beverly,MA),followed by incubation with bioti-nylated goat anti-rabbit IgG(BioCare,Concord,CA).Color was developed with Vectastain ABC(Vector Labs,Burlin-game,CA)and peroxidase substrate solution(Vector Labs) for10min,counterstained with hematoxylin.
Rat hind paws were snap frozen in liquid nitrogen and lysates immunoprecipitated(4μg paw lysate in10μl lysis buffer)with30μl of anti-Syk antibody conjugated to agarose beads(N-19,sc-1077AC,Santa Cruz Biotechnology Inc.,Santa Cruz,CA).Isolated beads were resuspended in loading buffer (Invitrogen,Carlsbad,CA),protein subjected to SDS-P AGE and transferred to PVDF membrane(Invitrogen,Carlsbad,CA). Membranes were incubated with primary anti-Syk monoclonal antibody ab3993(Abcam,Cambridge,MA),then secondary anti-mouse IgG-HRP antibody(Amersham,Piscataway,NJ),and developed with enhanced chemiluminescence(Amersham). Synovial fluid and synovium cytokine/chemokine analysis
On the day of sacrifice both hind legs of each rat were transected between the distal femur and proximal tibia. After removal of skin and hair,a2-mm incision was made lateral to the patella region of each leg.First,synovial fluid of the knee was collected by lavaging the viscous fluid under the patella and around the joints,followed by flushing two times,each time flushing with40μl of ice-cold PBS.The synovial fluid supernatant was collected and stored imme-diately at?80°C until multiplex immunoassay analysis. Synovium was collected by microdissection and immediately frozen in liquid nitrogen.Frozen synovial tissue was pulverized under liquid nitrogen,weighed,resuspended in RIPA buffer(250mg tissue/ml buffer),homogenized for1min at30,000rpm.Supernatants were centrifuged for2addi-tional cycles then diluted for immunoassay.Immunoassays were performed using a rat cytokine/chemokine multiplex immunoassay kit(Linco,St.Charles,MI)consisting of8 cytokines/chemokines(IL-1α,IL-1β,IL-6,IL-18,MCP-1,Gro/ KC,TNFα,and IFN-γ),and read on a Luminex100instrument (Luminex Corp.Austin,TX).The limits of detection were 15pg/ml for IL-1α,11.9pg/ml for IL-1β,14pg/ml for IL-6, 19.8pg/ml for IL-18,9.6pg/ml for MCP-1,1.6pg/ml Gro/KC, 8pg/ml for TNFα,and5.6pg/ml for IFN-γ.
Data analysis
Significant changes in clinical arthritis as a result of drug treatment were determined using a dynamic modeling approach, assuming a linear fit for the slope of arthritis progression for each individual animal(SAS Institute,Inc.,Cary,NC).A repeated measures analysis using an assumption of a constant correlation structure was used to corroborate the determination of statistical significance.Significant differences in serum or synovial cyto-kines,antibody levels,radiology scores,and histopathology scores were assessed using the Student's t-test applying the Welch's correction or the Wilcoxon rank-sum test,if applicable (GraphPad Prism3.0,San Diego,CA).
Results
R788/R406inhibit the Arthus reaction
Since FcγR(I,III)expressed on synovial macrophages are critical in the onset of IC-mediated inflammation,R406and R788activity was assessed in the RPA reaction.IC formation produced a severe Arthus reaction beginning within minutes after antigen/antibody injection,which increased in severity for4–8h.Prophylactic treatment of rats with R406or R788 administered1h prior to IC challenge resulted in a significant reduction(p b0.05)in vascular leakage(Fig.1),and the severity of local edema could be assessed by measurement of dye extravasation into the surrounding tissue.
R406treatment reduced the cutaneous RPA reaction and inflammatory edema in a dose-dependent manner,which was reflected by an approximate72%reduction of extravascular leakage of Evans blue dye and tissue swelling at the highest dose level used(10mg/kg)compared with the vehicle control (p b0.05).Treatment with vehicle alone had no effect on the RP A.The mean net OD610of extravasated blue dye into the surrounding tissue after treatment with10mg/kg R406was 0.05±0.01compared to0.17±0.02for animals treated with vehicle(p=0.004).
Similarly,treatment of rats with R788reduced the cutaneous RP A reaction and inflammatory edema in a dose-dependent manner(Fig.1B),which was reflected by an approximately70%and90%reduction of extravascular leakage of Evans blue dye and tissue swelling at the10mg/kg and 30mg/kg dose,respectively,compared with the vehicle control.In rats treated with R788,the mean net OD610 following treatment was reduced to0.05±0.005(10mg/kg, p b0.0001)and0.02±0.006(30mg/kg,p b0.0001).Four hours after challenge,the average net OD610from saline and vehicle-treated animals was0.24±0.02and0.3±0.04,via R406, respectively.This difference was not statistically significant. These findings showed that both the prodrug(R788),via R406, and the active metabolite(R406)directly administered could inhibit local inflammatory injury mediated by IC.
Oral administration of R788is metabolized to R406 The pharmacokinetic profile of the metabolite(R406)and the prodrug(R788)in both rat strains indicated that approximately equivalent systemic exposures of R406(AUC0–16)at a specified dose was obtained when Lewis rats were dosed twice daily and LOU rats were dosed once daily.The pharmacokinetic profile of R406in Lewis rats indicated that systemic exposure is proportional to dose,with an AUC0–16of7295ng h/ml for the10mg/kg dose and17,951ng h/ml for the30mg/kg dose. The C max of R406was1500ng/ml at10mg/kg(1h),and the measured peak at the high dose(30mg/kg)was delayed by approximately1h(4500ng/ml at2h).The pharmacokinetic profile of the prodrug,R788,in LOU rats indicated an AUC0–16 for systemic R406of10,618ng h/ml for a single10mg/kg dose
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and 30,650ng h/ml for a 20mg/kg dose.The C max was observed at 1h for the both doses (approximately 2600ng/ml at 10mg/kg and 6500at 20mg/kg)with a T 1/2of 4.2h.R406exposure was greater with the higher dose of R788,and the target concentration of R406(500ng/ml)was sustained for at least 8h in the LOU strain.The prodrug was not detected in the plasma in either rat strain,indicating that R788was completely converted to the active metabolite,R406.
Oral administration of R788/R406inhibits CIA progression and severity
Immunization with either bovine (in Lewis rats)or chicken (in LOUVAIN [LOU]rats)collagen type II (CII)co-administered with incomplete Freund's adjuvant (IFA)produces a severe
clinical arthritis beginning approximately 10–11days after immunization.R406or R788administration was initiated when at least one hind paw was inflamed (clinical arthritis score =1);enrollment into a treatment group was designated as day 0of the study for each individual animal.Progression of disease was evidenced by increased edema and erythema of one or both ankle joints,followed by involvement of the metatarsal and interphalangeal joints.Fully developed arthritis,including red and swollen paws,was observed 3–7days after onset of inflammation (maximum severity for LOU rats =6.1±0.7;maximum severity observed for Lewis rats =5.6±0.5).
R406or R788treatment of arthritic rats reduced the progression and severity of clinical arthritis that was evident within 7days after initiation of therapy,and the clinical condition of rats receiving drug therapy at efficacious doses remained improved throughout the study.In Lewis rats (Fig.2A),the high dose of R406(30mg/kg twice daily [b.i.d.])completely suppressed progression of arthritis in a majority of the animals (12/13),and the clinical arthritis scores regressed to “0”by the end of the study (mean clinical score =0.1±0.1,p b 0.0001vs.vehicle).Moderate arthritic symptoms were observed in only one paw of one rat treated with 30mg/kg b.i.d.(clinical score=2.5)for approximately 1week of the study (days 7–12),and by the end of the study,only mild redness was apparent in one of the paws.Treatment with the lower dose of 10mg/kg b.i.d.was also efficacious and resulted in a delayed onset and reduced clinical arthritis (mean clinical score =3.3±0.06,p b 0.0001vs.vehicle).The clinical score in the vehicle control group reached a peak approximately 1week after disease onset with a mean arthritis score of 5.6±0.5,and arthritis symptoms did not change during the study.Measurements of paw thickness and paw weight (data not shown)were consistent with clinical scores.Similar results (not shown)in LOU rats treated with R406were observed,with a highly significant reduction in arthritis severity (p b 0.001in rats treated with 30mg/kg R406once a day (qd)compared with vehicle).
Treatment of arthritic LOU rats with R788alone (15or 30mg/kg/day)significantly reduced clinical arthritis (p b 0.0001)compared with vehicle (Fig.2B).R788treat-ment,initiated after the onset of clinical arthritis,pre-vented an increase in arthritis scores in animals treated with 30mg/kg R788per day,and after 18days of treatment,the mean arthritis score was reduced to 2.4±0.2(compared with 7.4±0.6in vehicle-treated rats).R788administered at a dose level of 15mg/kg administered once a day (qd)reduced the mean clinical arthritis score to 4.1±0.6by day 18.In LOU rats the arthritic score gradually increased in the rats treated with vehicle alone after day 2,and by day 8,the mean arthritic score reached 7.2±0.6and remained essen-tially unchanged throughout the study.Lewis rats also responded to treatment with R788(data not shown).Thus,R406administered directly or indirectly as its prodrug (R788)regressed joint inflammation and edema in two rat models of rheumatoid arthritis.
The inhibition of FcR signaling following IC engagement should reduce the clearance and extend the half-life of antibodies administered therapeutically.Therefore,we examined the effect of R406on clearance of a commonly used biologic (anti-TNF αantibody)by administering
R406
Figure 1The reduction in severity of local edema with R406or R788prophylactic treatment was assessed by the concentration of Evans blue dye extracted from dissected tissues (OD 610).The concentration of blue dye,which reflected the extravascular leakage of dye and the severity of inflammatory edema,was calculated by subtraction of corresponding IgG control tissues from anti-OVA challenged sites.(A)Vascular leakage of dye in rats treated with R406.The percent inhibition in animals treated with 5or 10mg/kg,compared with vehicle control,was approxi-mately 60or 70%,respectively,compared with control and was statistically significant (*p b 0.05,**p b 0.01vs.vehicle).(B)The treatment with R788reduced vascular edema following treat-ment with 10mg/kg or 30mg/kg of R788,compared with control,and the percent inhibition was 83%or 93%,respectively (p b 0.001vs.vehicle).Data are presented as mean O.D.±SEM.
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(30mg/kg)in combination with a heterologous mouse monoclonal anti-human TNF αantibody.After a single dose of R406,mouse anti-human TNF αantibody levels were 33%higher in treated rats.The improved pharmacokinetic profile of R406that has been observed in normal human volunteers [20],suggests that co-administration of R788/R406with biological therapeutics may result in reduced clearance of the antibody therapies.
Bone destruction protection with R788/R406
The hind paws were examined by radiography and micro-CT .No evidence of bone damage was observed in X-rays obtained from LOU or Lewis rats treated with R788or R406(30mg/kg/day or 30mg/kg b.i.d.,respectively).At study termination
(day 18),using a score range of 0–6,with a score of “0”indicating no destruction and a score of “6”indicating severe destruction in both paws,blinded radiographs demonstrated a significant protection of joint damage in R406-or R788-treated rats (Fig.3).Arthritic Lewis rats treated with 30mg/kg R406b.i.d.had mean radiographic scores of 0.69±0.35(p b 0.0001vs.vehicle),compared with 3.69±0.60for rats treated with 10mg/kg b.i.d.R406(p b 0.02vs.vehicle),and 5.46±0.31for rats treated with vehicle alone (Fig.3A).LOU rats (Fig.3B)treated with R788(30mg/kg qd)had mean radiographic scores of zero (p b 0.00000001vs.vehicle).None of the rats treated with the high dose (30mg/kg qd)of R788(11/11)had any signs of bone destruction.Rats treated with the intermediate dose of 15mg/kg qd R788had mean X-ray scores of 0.6±0.3,p b 0.000007vs.vehicle,while those treated with vehicle alone had mean radiographic scores of 5.1±0.6,indicating that treatment with R788was highly effective in protecting bone integrity.
Photographs of rat limbs on day 18showed that no edema or inflammation was observed in hind paws of treated rats (Fig.4,upper panel).In immunized rats treated
with
Figure 3Blindly scored radiographs demonstrated a signifi-cant reduction in bone erosion with animals treated with (A)R406(10mg/kg b.i.d.,p b 0.05vs.vehicle;30mg/kg b.i.d.,p b 0.001vs.vehicle)and (B)R788(p b 0.001,15and 30mg/kg qd vs.vehicle).Data are the means ±standard error for n =13per
group.
Figure 2(A)Mean hind paw clinical scores ±standard error as a function of time.Disease onset (day 0)occurred when a rat was scored a “1”or more on one or both hind paws.Lewis rats were treated with vehicle or R406(10and 30mg/kg)at disease onset.Arthritis clinical scores from both hind paws of each rat were evaluated daily during the 18-day treatment course.Oral treatment of R406at 10or 30b.i.d,significantly ameliorated the severity and development of arthritis (p b 0.0001,10mg/kg and 30mg/kg vs.vehicle).(B)LOU rats were immunized with chick CII/IFA.Rats (9–11per group)were randomized into treatment groups on the day of arthritis onset (day =0).Clinical arthritis was scored daily.Oral treatment of rats with R788once a day (15or 30mg/kg)reduced clinical arthritis (p b 0.0001vs.vehicle).
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Syk inhibitor suppress C/A
vehicle alone,edema and inflammation were evident in the hind paws (Fig.4A).Administration of R406inhibited the development of arthritis,indicated by the absence of paw swelling or edema and inflammation in hind paws (Fig.4B).A representative X-ray of a paw from a rat treated with vehicle alone is shown in Fig.4C,and an X-ray of a rat treated with R406(30mg/kg)is shown in Fig.4D.Radiographic evidence of disintegration of cortical surface in the vicinity of the inflamed joints is clearly apparent in the arthritic rats treated with vehicle alone,whereas bone is preserved,or bone destruction retarded,in rats treated with R406.Notably ,the joint deformity observed in control rats was completely eliminated,leaving the appearance of paw damage indistinguishable from that of naive animals.Micro-CT analysis of the hind limbs of arthritic LOU rats treated with vehicle alone showed a substantial loss of bone integrity and damage,whereas animals treated with 30mg/kg R788qd showed no evidence of bone erosion (Fig.5).
Reduction of cartilage degradation products in R788/R406-treated rats
The appearance of cartilage oligomeric matrix protein (COMP),a cartilage degradation product in the serum,is associated with loss of bone integrity in RA patients [28,29].In response to treatment of arthritic rats with either R406or R788,serum COMP was significantly reduced in treated animals (Fig.6).The net average serum COMP levels in Lewis rats treated with 10mg/kg or 30mg/kg R406were decreased 53%and 63%respectively when compared with the vehicle group.Similarly,R788reduced serum COMP levels in Lewis rats (1.5±0.2U/l,p b 0.001,at a dose level of 10mg/kg b.i.d.),compared with those treated with
vehicle alone (serum COMP level =2.7±0.22U/l).Overall,treatment of arthritic rats with either R406or R788reduced serum COMP levels in a dose-dependent manner ,although a plateau was reached.Interestingly,R788and R406com-pletely prevented any evidence of bone destruction by plain radiographs or micro-CT ,suggesting that subclinical carti-lage degradation occurred in these studies or that
R788/
Figure 4Upper panel:representative photographs of hind paws from a control rat (A)and a treated rat (B)revealed marked swelling and redness with joint thickening that was undetectable following treatment with R406.Arrows indicate swollen and erythematosus regions (A)and no swelling (B).Lower panel:radiographs of hind limbs were obtained at the conclusion of the study.Representative radiographs from a LOU rat treated with vehicle (C)and 30mg/kg R406(D).Bone erosion is evident in animals treated with vehicle alone;circled areas indicate soft tissue swelling and erosions at the ankle and tarsus.No evidence of any damage is seen in animals treated with R406(circled areas highlight the absence of swelling and bone erosions).Radiographs of paws from LOU rats are indistinguishable from paws of Lewis rats (X-rays not
shown).
Figure 5Micro-CTanalysis of hind limbs excised from arthritic LOU rats treated with vehicle alone (A)or R788(B)for 18days.Treatment of rats with R788reduced the bone loss seen in arthritic rats.The integrity of the bone structure is maintained (circled areas)in animals treated with R788.
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R406treatment reduced,but did not eliminate,cartilage turnover .
Reduced histologic synovitis and destruction in R788/R406-treated rats
Hind paws evaluated for histopathological changes in cartilage structure,cellularity,matrix glycosaminoglycans,and synovial inflammation indicated that arthritic rats exhibited extensive inflammatory infiltrate within the synovium and synovial hyperplasia.Erosion of articular cartilage in animals treated with vehicle alone (Figs.7A and B)was most severe at the joint margins,sometimes extending to the subchondral bone,and glycosaminoglycan content of the cartilage was moderately to severely reduced in CIA.Hind paw sections from rats treated with R406indicated a reduction of synovitis that was also seen in rats treated with R788(Figs.7C and D).
Composite histopathology scores from rats treated with either R406or R788are presented in Fig.7E,and the scoring system is presented in Fig.7F (0–17for each hind paw,with a score of “0”indicating no damage,and a score of “17”indicating complete disorganization and loss of matrix glycosaminoglycans,tissue necrosis,marked inflammation and hyperplasia).The high dose of R406(30mg/kg b.i.d.)reduced modified Mankin scores,with an approximate equal reduction in severity for all parameters examined (2.0±3.9,p b 0.0001vs.vehicle),while treatment with R406at an intermediate dose (10mg/kg b.i.d.)significantly reduced mean scores to 8.9±5.6(p b 0.001vs.vehicle),compared with a total score of 14.4±3.1in animals treated with vehicle alone.Treatment of Lewis rats with R788reduced histo-pathological scores and synovial inflammation;an inter-
mediate dose of 10mg/kg R788b.i.d.significantly reduced inflammatory changes in the joints (mean score of 7.2±6.6,p b 0.001vs.vehicle),while a low dose (5mg/kg b.i.d.)resulted in a modest impact on joint integrity (mean score of 12.0±3.5,p N 0.05vs.vehicle).The mean total score in animals treated with vehicle alone was 15.4±1.7.
Expression of Syk in tissue sections from vehicle-treated rats (Figs.8A –C)and rats receiving R406(Figs.8D –F)correlated with the extent of inflammation.Joint sections from animals treated with vehicle and stained with anti-Syk antibodies,H&E,or Safranin O,showed prominent areas of Syk expression (Fig.8A),narrowing of the joint space and extensive inflammatory cells in the pannus (Fig.8B),and severe reduction of proteoglycans (Fig.8C),whereas in animals treated with R406(R788),Syk expression was nearly undetectable (Fig.8D),inflammation was substantially reduced (Fig.8E),and synovial glycosaminoglycans were better preserved (Fig.8F).Western blot analysis of paw Syk protein (Fig.9)demonstrated that Syk expression following treatment with R788/R406was reduced to levels of that seen in naive rats.Modulation of Syk expression is most likely due to an overall dampening of the inflammatory cascade and reduced inflammatory cell infiltration into the joint tissues following R788/R406treatment.
Effects on cytokine and chemokine levels
The cytokine and chemokine profile from synovial fluid and synovial tissue was examined in rats treated with R406(Lewis)and R788(LOU),respectively.Tissues and fluid were collected at sacrifice (18days after the start of drug treatment).Eight cytokines/chemokines (Gro/KC,IFN-α,IL-18,IL-1β,IL-6,IL-1α,MCP-1and TNF α)were analyzed using a multiplex immunoassay.IL-18,IL-6,MCP-1,and IL-1β,and Gro/KC were consistently and significantly increased in vehicle-treated arthritic rats compared to naive rats (Fig.10),and treatment with R406completely suppressed the expression of these 5cytokines and chemokines in arthritic rats (R406,30mg/kg b.i.d.).A similar pattern of cytokine expression was seen in synovial fluid isolated from the ankle joints (unpublished data).Synovium was microdissected from the ankle joints of LOU rats treated with R788or vehicle.Most of the eight cytokines were below the limit of detection;however ,measurable levels of IL-18and Gro/KC were demonstrated in the synovium.Treatment of LOU rats with R788(30mg/kg qd)resulted in an approximate 50%decrease of IL-18(2896.5pg/ml in vehicle-treated rats vs.1288.3pg/ml in R788-treated rats)and Gro/KC (147pg/ml in vehicle-treated rats vs.93pg/ml in R788-treated rats).
Disease attenuation is not attributable to inhibition of humoral collagen-specific immunity
Sera were analyzed at various times after immunization and at sacrifice for anti-CII antibody by an ELISA (Fig.11).Sera from Lewis rats were collected 2days prior to initiation of drug treatment and on day 7after the onset of disease,as well as at the time of sacrifice (day 18).The CII IgG antibody levels in the sera of rats increased as disease progressed,and the level of serum CII antibody was highly elevated prior to any signs of joint inflammation or tissue swelling.
There
Figure 6Prevention of cartilage destruction in rats treated with R788/R406.The average COMP in rats treated with vehicle alone was 2.55±0.19U/l.(A)After 18days of treatment,R406administered at dose levels of 10mg/kg or 30mg/kg significantly reduced COMP levels (p b 0.001vs.vehicle)to mean levels of 1.62U/l and 1.45U/l in animals treated with 10or 30mg/kg R406,respectively ,compared with mean serum COMP of 2.55U/l in rats treated with vehicle.(B)R788treatment reduced serum COMP to 2.1U/l (5mg/kg b.i.d.,p N 0.05)and 1.5U/l (10mg/kg b.i.d.,p b 0.001),compared to 2.7U/l in rats treated with vehicle alone.
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was no statistically significant difference in CII antibody (p N 0.05)between R406-treated groups compared to the vehicle control at any time point tested.Likewise,sera collected from LOU rats treated with R788at study termination indicated that treatment did not change anti-body level.The mean CII antibody level (O.D.)in
arthritic
Figure 7Histopathology of hind paw sections from Lewis rats treated with R788/R406.Note changes in cellular infiltration,Safranin O staining,and cartilage erosions.(A)Vehicle 20×,(B)vehicle 100×,(C)R40630mg/kg qd 20×,(D)R40630mg/kg qd 100×.(E)A quantitative assessment of histopathological hind paw sections for the changes in cartilage structure,cellularity,matrix glycosaminoglycans,and synovial inflammation indicated a significant reduction in tissue destruction was seen in rats treated twice daily with R406or R788.Data are the means ±standard error for n =13per group.(F)The detailed tabular summary of the scoring system used to evaluate the reduction in joint destruction.
252P .R.Pine et al.
Figure 8Syk expression in hind limbs.(A)Vehicle,anti-syk;(B)vehicle,H&E;(C)vehicle,Safranin O;(D)R40630mg/kg b.i.d.,anti-syk;(E)R40630mg/kg b.i.d.,H&E;(F)R40630mg/kg b.i.d.,Safranin O.All magnifications are 100×.Note the increased Syk expression,inflammatory cell infiltrate,and joint-space narrowing in joints of arthritic rats treated with vehicle only (A –C),which is reduced in animals treated with R406(D –F)or R788(data not shown).Tarsus sections were deparaffinized,blocked with 1.5%normal goat serum (Vector Labs,Burlingame,CA)for 1h,then incubated with polyclonal antibody to Syk (Cell Signaling,Beverly,MA)for 1h,washed and incubated with biotinylated goat anti-rabbit IgG (BioCare,Concord,CA).Sections (A,D)were treated with Vectastain ABC (Vector Labs,Burlingame,CA)and peroxidase substrate solution (Vector Labs)and counterstained with hematoxylin.Slides were stained with hematoxylin and eosin alone (B,E),or Safranin O (C,F).
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rats treated with vehicle was 0.195±0.001compared with 0.191±0.002for rats treated with 15mg/kg R788qd (p =0.12)and 0.193±0.004for rats treated with 30mg/kg R788qd (p =0.68).In the short-term treatment model used in these studies,blockade of B cell function and decreased autoantibodies may not be immediately detectable,since the in vivo half-life of immunoglobulins can extend several weeks.
Discussion
The results reported here demonstrate for the first time that inhibition of Fc γR signaling via Syk can completely suppress joint inflammation and bone erosion in CIA.The pharmaco-logical anti-inflammatory potential of R788,a prodrug of R406,to inhibit activating FcR signaling was initially demonstrated in a cutaneous reverse passive Arthus reac-tion,in which inhibition of FcR signal transduction reduces the IC-mediated inflammatory response [30,31].In the CIA model,treatment of arthritic rats with R788/R406,adminis-tered after disease onset,completely suppressed all mani-festations of arthritis in rats,including tissue swelling,joint space narrowing,inflammatory cell infiltration into the synovial fluid and tissue,inflammatory cytokine secretion
into the joint space,IL-18accumulation in the synovium,bone erosion,and accumulation of cartilage degradation products in the https://www.doczj.com/doc/9f17647363.html,ing immunochemistry,the expres-sion of Syk in joint tissue was reduced in animals treated with R406/R788;the reduction of Syk expression in tissues may be due to the inhibition of inflammatory cell infiltrates as a result of R788/R406treatment.Pharmacokinetic analysis demonstrated that R788is rapidly and completely converted to R406in the gastrointestinal tract,providing an orally bioavailable small molecule alternative and/or addition to protein therapeutics.
In RA patients,as well as in the synovium of rats immunized with CII and adjuvant,inflammatory disease is marked by synovial hyperplasia associated with neutrophil and macrophage infiltration,increased proinflammatory cytokines,and inflammatory cell invasion of bone and cartilage [32,33].Following sensitization of rodents (active or passive),macrophage-like phagocytic cells line the synovium and synovial-like fibroblasts in the sublining region proliferate,resulting in massive recruitment and
infiltration
Figure 9(A)Pulverized hind paws from naive,vehicle-,or R406-treated rats were probed with monoclonal anti-Syk using Western blot analysis.(B)Pulverized hind paws from either naive,vehicle-,or R406-treated rats were immunoprecipitated with polyclonal anti-Syk agarose beads.Western analysis of immunoprecipitated samples was probed with
anti-Syk.
Figure 10Synovial fluid of animals treated with vehicle alone showed elevated levels of Gro-KC,IL-1β,IL-18,MCP-1and IL-6that were reduced in treated rats to the levels seen in naive rats (black bar).Rats were treated with either vehicle (white bar)or R406(grey bar)for 18days (30mg/kg b.i.d.),synovial fluid was lavaged from the knee area at study termination and analyzed using multiplexed
immunoassays.
Figure 11Serum was collected on day ?2(2days prior to disease onset),day 8(1week after the onset of clinical arthritis in the majority of animals),and day 18after the start of drug treatment.Anti-CII was elevated as disease progressed;treat-ment with R788/R460did not reduce antibody levels (see text).Data are the means±standard error for vehicle (n =8),10mg/kg (n =7),30mg/kg (n =8),and naive (n =3)groups.
254P .R.Pine et al.
of neutrophils and macrophages.In addition to synovial hyperplasia,bone destruction is a predominant feature of RA associated with pain and morbidity.Erosion of trabecular and cortical bone is common,leading to the characteristic erosions seen on radiography[34].Notably,our studies demonstrated that no radiographic damage whatsoever was observed in rats treated with R788/R406.Micro-CT analysis and a reduction of serum COMP levels in treated rats support the protective effect of R788/R406treatment in the reduction of bone destruction.In patients with RA,elevated serum COMP levels are an important marker of cartilage degradation;treatment with TNFαinhibitors has been shown to reduce COMP levels[35],suggesting the possibility of using this biomarker to follow disease progression.
As bone erosion results from the proliferation and activation of osteoclasts,complete inhibition of bone erosion in joints of rats treated with R788/R406may result from R406 action on the osteoclast itself.Mocsai et al.[36]have shown that Syk is critical in regulating the development of functional osteoclasts through DAP12and FcRγ.Both DAP12and FcRγcontain ITAM domains that couple to downstream signaling pathways through Syk.In addition,DAP12?/?,FcRγ?/?,and syk?/?osteoclasts fail to resorb mineralized matrix.While most anti-rheumatic drugs such as methotrexate,corticoster-oids,and nonsteroidal anti-inflammatory drugs can suppress inflammation and pain,only a few agents are capable of slowing bone and cartilage erosion.In particular,the benefits of Anikinra[37],leflunomide[38],and anti-TNFαtherapy administered in combination with methotrexate[39,40]on bone structure have been reported.
The primary consequence of R788/R406treatment does not appear to be on B cell function in the rat CIA model. Treatment of arthritic rats with R788/R406did not significantly alter serum CII antibody levels throughout the course of the study.CII antibody levels had increased to nearly maximum levels prior to the appearance of paw swelling and reached a plateau within4days following the onset of disease.R788/R406might still have a role in modulating autoantibody production,but the effect may be difficult to evaluate in this model,as immunoglobulin half-life can be several weeks.Targeted disruption of Syk has revealed important functions of Syk in immune cell development and in B cell receptor signaling[41,42],and disruption of B cell function will likely be beneficial to RA patients.
Inhibition of Syk in other inflammatory cell types, including antigen-presenting cells,mast cells,neutrophils, macrophages,and B cells or in non-inflammatory cells,such as synoviocytes[23]and osteoclasts[36],contribute directly to the protective effects of R788/R406in CIA.Impaired dendritic cell(DC)migration to the draining lymph nodes, resulting in defective DC maturation,antigen presentation, and T cell activation[43,44]supports the potential of FcR-mediated modulation of DC maturation and antigen pre-sentation as a therapeutic strategy for the treatment of RA [45].R788/R406may exert indirect effects on T cell phenotypes through its modulation of antigen presentation or B cell activation.Interestingly,an aberrant T cell signaling phenotype has been reported in patients with systemic lupus erythematosus(Liossis et al.[46])in which the TCR activation occurs through FcγR and Syk,rather than through the canonical TCR activation pathway involving the zeta chain and Zap-70,suggesting that R788/R406may be particularly useful for the treatment of lupus or other autoimmune disorders in which this aberrant phenotype may be found.To our knowledge,this phenotype has not been described in RA patients.Recent observations by Cha et al.[23]show that Syk expression is increased in synovial tissue of RA patients,and that R406inhibited TNFα-induced Syk activation in cultured FLS and decreased production of IL-6and MMP.
R406is relatively selective to Syk inhibition as assessed using anti-phosphopeptide Western blots,although we have demonstrated that R406also inhibits Flt3,Jak,and Lck, albeit less potently than Syk[20].R788/R406treatment reduces activated T lymphocytes and the number of class-switched germinal center B cells,as well as selectively reduces costimulatory molecules in non-B-cell antigen-presenting cells in a spontaneous model of SLE(unpublished data).Thus,inhibition of pathways other than Syk may be responsible for the potential effects of R406in CIA.
Modulation of proinflammatory cytokines,specifically IL-1and TNFα,are known to improve clinical outcomes in RA.In patients,elevated level of IL-1βhas been characterized as one of the manifestations of bone resorption in RA[39].In addition,local overexpression of IL-18,a member of the IL-1 superfamily,resulted in pronounced joint inflammation and stimulated the release of proteoglycans from human articular cartilage[47].IL-6has been shown to be associated with osteoclastogenesis,and blockade of these cytokines reduces bone resorption[48].In animal models,overexpres-sion of IL-18[49]in TNF-deficient mice exacerbated cartilage proteoglycan loss,compared with mice expressing TNF.IL-1βblockade prevented cartilage and bone destruc-tion in mouse CIA,whereas TNFαblockade only ameliorated joint inflammation[50].In the study reported here, proinflammatory cytokines,Gro/KC,IL-6,IL-18,IL1β,and the protease,MCP-1,were highly elevated in the synovial fluid isolated from arthritic rats,and these inflammatory mediators were significantly reduced following treatment with the Syk inhibitors.TNFαwas not detected in our assays, likely due to the time of sampling,as anti-TNFαtreatment in the mouse CIA model is effective only if started shortly after disease onset.The reduction of synovial cytokines is consistent with the effect on disease severity reported here for R788/R406in the CIA model.
The successful treatment of RA sufferers with biological therapies,including TNFαinhibitors[51],IL-1receptor antagonist[39],anti-CD-20monoclonal antibody[52,53], and the costimulator antagonist,CTLA4Ig[54–57],and the capacity of R788/R406to decrease the clearance of IgG antibodies,as reported here,suggests that R788might also be useful in extending the half-life and efficacy of these Fc-containing biologic agents and other antibody therapies.
In conclusion,we have shown that the pharmacological intervention of FcR signaling via Syk inhibition,can effec-tively ameliorate CIA.Clinical trials(phase II)are currently underway to test the hypothesis that Syk is a therapeutic target for autoimmune diseases,including RA. Acknowledgments
The authors gratefully acknowledge Abraham Silvers for the statistical evaluation of clinical arthritis,Muhammad Baluom
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for the pharmacokinetic analysis,and Anne Ponugoti for graphics assistance.
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Syk inhibitor suppress C/A
中英文自荐信范文 第一篇：中英文-自荐信dear sir, professor kenneth h. chan, head of the department of metallurgy, has informed me that your agency is looking for an engineering graduate with the ability to write about technical subjects for non-technical readers. the four years at hong kong university have given me a thorough foundation in such fundamental sciences as physics, mathematics, and chemistry, in addition to specialized courses in metallurgy. i have also taken as many courses in english and psychology as possible with the expectation that i would enter the field of technical sales or editing following my graduation this summer. fortunately, as my resume shows, i now have a mature interest in finding a satisfying job. may i come in for an interview at your convenience? sincerely yours, 敬啟者: 冶金系主任陈教授告诉我,贵公司在物色有能力写专门性文章给一般读者阅读的工科毕业生。 在香港大学四年我奠定了冶金及物理、数学、化学基本科学的良好基础。我曾经选修许多门英语和心理学方面的课程,期望今年夏天
电商常用术语中英对照 ?CVR (Click Value Rate): 转化率，衡量CPA广告效果的指标 ?CTR (Click Through Rate): 点击率 ?CPC (Cost Per Click): 按点击计费 ?CPA (Cost Per Action): 按成果数计费 ?CPM (Cost Per Mille): 按千次展现计费 ?CPS(Cost Per Sales)：以实际销售产品数量来换算（淘客） ?PV (Page View): 流量 ?PV单价: 每PV的收入，衡量页面流量变现能力的指标 ?ADPV (Advertisement Page View): 载有广告的pageview流量 ?ADimp (ADimpression): 单个广告的展示次数 ?RPS (Revenue Per Search): 每搜索产生的收入，衡量搜索结果变现能力指标 ?ROI：return on investment回报率(ROI)是指通过投资而应返回的价值，它涵盖了企业的获利目标。利润投入的经营所必备的财产相关，因为管理人员必须通过投资和现有财产获得利润。又称会计收益率、投资利润率。?UV(独立访客)：Unique Visitor ?PV(访问量)：PageView,即页面浏览量或点击量，用户每次刷新即被计算一次。 ?SKU：SKU=Stock Keeping Unit（库存量单位）
销售常用术语及释义1.时间 2.生意衡量指标 1) 销售指标 ?GSV Gross Sales Value Sell-in的销售额 ?NSV Net Sales Value Sell-in的销售额减去TP
眼镜行业市场现状和发 展趋势分析 集团标准化工作小组 #Q8QGGQT-GX8G08Q8-GNQGJ8-MHHGN#
中国眼镜行业市场现状和发展趋势分析 改革开放以来，我国眼镜行业所有制结构发生了很大变化，形成国有、集体、私营、合资、独资等多种所有制并存的局面，企业充满活力，眼镜行业从一个名不见经传的小行业，发展成一个充满生机，充满潜力的朝阳行业，我国也逐步发展成为世界主要眼镜消费国家和生产国家之一。目前中国不仅是世界潜力最大的眼镜消费大国，而且已成为世界领先的眼镜生产大国。 中国眼镜协会分析，近5年来，中国眼镜产业年均增幅达17％。目前，我国眼镜生产企业超过4000家，有一定规模的验光配镜店超过2万家，并已形成广东东莞、福建厦门、浙江温州、江苏丹阳、上海、北京等主要生产基地。眼镜的材质、品种、款式日益多元化，能够生产纯钛、钛合金、记忆金属等眼镜架、角膜接触镜、CR-39光学树脂镜片、镀膜镜片、渐进多焦点镜片、非球面镜片等高技术含量的新型眼镜产品，并出现江苏“万新”、浙江“海豚”等知名品牌。据保守估计，中国约有3亿人配戴眼镜（矫正屈光镜），按每3年更新一付来计算，每年的市场需求量就达1亿付，如果加上太阳镜，市场需求量就更为可观。除了实用功能，眼镜还被赋予装饰功能，眼镜消费的个性化、时尚化、品牌化、高档化的趋势日益显着，这进一步催生了中国眼镜市场的商机。 受中国市场诱惑，几乎所有的国际知名眼镜厂家和商家纷纷来华掘金，或设立生产基地，或编织销售网络。譬如，1997年，全球最大的镜片制造商法国依视路(ESSILOR)投资5300万美元在上海建立了镜片生产基地，产量每年以两位数的速度增长，并占据了12％的中国市场。日本野尻1989年登陆中国，不仅在上海设立了镜架生产厂，而且在全国布局了上千家连锁店。香港是全球第二大眼镜生产中心，目前已将70％－80％的生产企业转移到广东东莞。台湾是全球主要的太阳镜生产中心，也将生产重心转移到福建厦门。。
中英文自荐信范文 第1篇第2篇第3篇第4篇第5篇更多顶部 目录 第一篇：中英文-自荐信第二篇：中英文自荐信四第三篇：中英文自荐信五第四篇：个人简历中英文自荐信第五篇：医药学毕业中英文自荐信更多相关范文 正文第一篇：中英文-自荐信dear sir, professor kenneth h. chan, head of the department of metallurgy, has informed me that your agency is looking for an engineering graduate e a thorough foundation in such fundamental sciences as physics, mathematics, and chemistry, in addition to specialized courses in metallurgy. i have also taken as many courses in english and psychology as possible y graduation this summer. fortunately, as my resume shoature interest in finding a satisfying job. may i come in for an intervie the university of kansas asters degree in june of this year. my major is pharmacy and i am very interested in securing a position before i graduate, if possible. i am a hong kong chinese and my bachelor’s degree is
外文文献及译文 文献、资料题目：Electronic Commerce 外文文献： Electronic Commerce Electronic commerce, or Electronic trade, or electronic business as a newly rising mode of commerce will have far-reaching influence on social economy and play an important role in social development worldwide. It represents the trend of world trade in the 21st century and beyond. 1. What is Electronic Commerce? Electronic commerce refers to commercial data exchange in digital form through electronic transmission means and commercial activities conducted on-line. Usually, electronic commerce can be divided into two levels: One is low-level electronic commerce that is, electronic commercial intelligence, electronic trade, and electronic contracts. Another is high-level electronic commerce which includes all commercial activities done via Internet, ranging from searching for clients, commercial negotia-tion, making orders, on-line payment, releasing electronic invoice, to electronic dec-laration to Customs, electronic tax-payment, all conducted on Internet. Electronic commerce means electrification of all trade transactions. It is featured by these characters: ①fairness and freedom, ②high efficiency, ③globalization, ④virtualization, ⑤interactivity, ⑥autonomy, ⑦personalized service. With electronic commerce, clients and
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眼镜行业产业链及主要企业分析 眼镜是以矫正视力或保护眼睛而制作的简单光学器件，由镜片和镜架组成。眼镜既是保护眼睛的工具，又是一种美容的装饰品。中国庞大眼镜市场且增长潜力巨大，预计2022年中国眼镜行业销售收入将达547亿元，未来行业兼并整合将增多，企业正探索线上线下互动的经营模式。 眼镜零售行业现状 1、眼镜零售向连锁化发展 上世纪80年代之前，中国没有严格意义上的眼镜零售市场；80年代末和90年代初，我国眼镜店的结构中，以国营店为主，私营店处于萌芽状态，眼镜店主要销售的产品为玻璃镜片和款式单一、材质普通的镜架。随着金属钛架和树脂镜片的应用，新科技与营销技术带动了眼镜零售店的发展。 90年代中期，眼镜行业进入了快速成长期。在这期间，一些重视信誉的眼镜店在竞争中逐渐发展壮大，由单个店面发展壮大成为区域性或者跨区域的大型连锁店。如今，国内眼镜连锁企业如宝岛眼镜等也各自发力，规模逐渐成长，行业梯队逐渐形成，品牌企业开始涌现。至今，国内具有一定规模的眼镜零售店已遍布全国大中城市，并朝现代营销、连锁经营的方向发展。 2、多元化零售业态正在形成 随着居民可支配收入的日益上涨，消费者消费能力增强，消费心理也日渐成熟，眼镜的消费需求正朝着多元化、个性化的方向发展。因此，深度挖掘特定群体的消费偏好，细分消费者市场，已成为眼镜零售新趋势：既有中高端眼镜连锁品牌来满足中高端消费者的个性化需求，也有以价格为导向的平价眼镜超市以刺激低端消费；既有眼科医院等专业医疗视力保健机构，也有鼓励一人多镜的快时尚眼镜店。 3、外资企业进入国内眼镜零售市场 鉴于中国眼镜市场具有较大的发展潜力，一些国外眼镜零售巨头积极拓展国内市场。凭借着雄厚的资本，跨国眼镜公司纷纷通过投资、收购等方式抢滩中国市场。2005年，荷兰眼镜投资公司HALInvestments Asia .以亿元人民币收购上海老字号红星眼镜公司78%的股权，红星眼镜公司在全国拥有49家直营店和64家加盟连锁店。2005年7月，Luxottica收购北京雪亮眼镜，收购价为收购广东明廊眼镜，收购价亿元，收购经营网点278个；2006年11月，Luxottica收购上海现代光学，收购价亿元，收购门店28家。这些举措虽然在短时间内使外资企业的门店数量能迅速增长，但国内消费者消费理念、消费习惯、消费水平与国外消费者存在较大差异，进一步适应本土化的需求是外资企业提高在中国市场的竞争力的关键。 4、市场潜力巨大 我国经济的快速发展、国民收入和消费水平的不断提高，近视低龄化、近视人口的增加，社会老龄化程度的提高以及国内消费者保健意识的增强，催生中国庞大眼镜
【中文名】软骨发育不全 【英文名】achondroplasia 【定义】软骨发育不全是由于基因突变或者环境影响而导致的长骨生长增殖缺陷，主要表现为四肢短小，是最常见的一种侏儒症。 【临床表现】本病是侏儒的最常见原因。胎儿娩出时即可见其身体长度正常而肢体较短，这种差别以后逐渐明显。肢体近端如肱骨及股骨比远端骨更短。患儿脂肪臃肿，至发育成熟，平均身高男性为135cm，女性为123cm。文献上有身高62.8cm的报道。患儿身体的中点在脐以上，有时甚至在胸骨下端，两手只能碰到股骨下粗隆的下方，而不像正常人那样可以达到大腿下1/3。 头颅增大，有的患者有轻度脑积水，前额突出，马鞍型鼻梁，扁平鼻、厚嘴唇，舌伸出（婴儿）。中耳炎，反复发作会导致传导性耳聋。胸腔扁而小，肋骨异常的短。胸椎后突，腰椎前突，以后者为明显。骶骨较水平使臀部特征性的突出。手指粗而短，分开，常可见4、5指为一组，2、3指为一组，拇指为一组，似“三叉戟”。有的患者伸肘动作轻度受限。下肢呈弓形，走路有滚动步态。 智力发展正常，牙齿好，肌力亦强，性功能正常。 【诊断】诊断依据：①四肢短小与躯干长短不成比例，呈软骨发育不全性侏儒形，即头大、鼻塌、下颌及前额突出、颈短，臀部后突、腰前凸，骨盆前倾。②智力、性征和肌肉发育均正常。③手指呈车辐样散开，手常不能摸及臀部。④有的表现出脊柱侧弯，甚至出现椎管狭窄、脊髓和神经症状。常伴有O型腿畸形。 检查方法：产前超声检查及DNA检测、X线检查。 颅盖大，前额突出，顶骨及枕骨亦较隆突，但颅底短小，枕大孔变小而呈漏斗型。 长骨变短，骨干厚，髓腔变小，骨骺可呈碎裂样或不整齐。在膝关节部位，常见骨端呈V 形分开，而骨骺的骨化中心正好嵌入这V形切迹之中。由于骨化中心靠近骨干，使关节间隙有增宽的感觉。下肢弓形，腓骨长于胫骨，上肢尺骨长于桡骨。 椎体厚度减少，但脊柱全长的减少要比四肢长度的减少相对少很多。自第一腰椎至第五腰椎，椎弓间距离逐渐变小。脊髓造影可见椎管狭小，有多处椎间盘后突。 骨盆狭窄，各个径均小，髋臼向后移，接近坐骨切迹，有髋内翻，髋臼与股骨头大小不对称。肋骨短，胸骨宽而厚。肩胛角不锐利，肩胛盂浅而小。 【中文名】季肋发育不全 【英文名】hypochondroplasia 【定义】季肋发育不全是一种遗传病，患者四肢短小，前额突出，脊柱发育不正常，但症状通常于2～4岁后才会出现。 【临床表现】出生时症状并不明显，身高、体重及头围皆为正常。童年时期2～4岁时症状会变得显著，手、脚和四肢短小且不成比例，前额通常会突出但头颅部却并未受影响，脊柱有些不正常与软骨发育不全症相同，但依个案状况程度不同，较少像软骨发育不全症发生的情况。 季肋发育不全是一种不太严重的软骨发育不全症，通常患者成长至成人时身高为128～165cm。有些患者在童年早期会有典型的弓型腿发育，但随着年龄增长这种状况时常会自发性地改善，也有些患者会因此症而合并有巨脑症，此外约有10%的患者有中度的智能迟缓现象产生。 与正常人一样的生命期，可能会有中度的智力缺损。
应聘秘书的中英文求职信范文 Model Chinese and English application letter for Secreta ry
应聘秘书的中英文求职信范文 前言：个人简历是求职者给招聘单位发的一份简要介绍，包括个人的基本信息、过往实习工作经验以及求职目标对应聘工作的简要理解，在编写简历时，要强调工作目标和重点，语言精简，避免可能会使你被淘汰的不相关信息。写出一份出色的个人简历不光是对找工作很有用处，更是让陌生人对本人第一步了解和拉进关系的线。本文档根据个人简历内容要求和特点展开说明，具有实践指导意义，便于学习和使用，本文下载后内容可随意调整修改及打印。 英文版 personal details name: miss. liang xiaoyan nationality: china （mainland） current place: guangzhou height/weight: 163 cm marital status: married age: 27 years career objective
application type: jobseeker preferred job title: admin staff/assistant: assistant 、 shipping: 、 secretary/paper staff: working life: 3 title: no title job type: full time expected start date: in a day expected salary: ￥2，000--￥3，500 preferred working place: guangzhou work experience company's name: guangzhou sanny import and export co.， ltdbegin and end date: XX-01-XX-08 enterprise nature: private enterprisesindustry: mechnical follow all the business for old customers. 2> place new order to factory， follow producing process. 3> prepare paper work for custom declaration and inspection.
第三节治疗依从性评估 治疗依从性是指患者对医嘱治疗措施的遵从执行程度，可分为完全依从、部分依从和完全不依从3类。治疗依从性不良在精神疾病患者中较为普遍，可引起多方面的后果，包括治疗失败、病情难以控制、反复住院、不良反应增加等，从而导致病程延长，严重影响生活质量与身体健康，同时也导致医疗费用的增加。因此应重视治疗依从性评估，采取积极有效的措施提高患者的治疗依从性。 治疗依从性没有统一适用的评估标准和形式，主要采用的方法有等级法（依据程度分为“好”、“中等”、“差”、“过度依从”）、自知力、药物用量计算法、药物浓度监测、依从性量表等。本节结合以上方法进行综合评估，对涉及药物评估的内容请参考第八节。 [目的] 1．监测患者对治疗的依从程度，识别出依从性不良患者。 2．根据依从程度采取针对性干预措施，提高治疗和康复的效果。 [用物]病历资料、各种治疗登记表、自知力与治疗态度问卷等。 [操作步骤] 1．环境准备安全、舒适。 2．护士准备着装整洁、情绪饱满、熟悉评估内容和流程，准备评估相关物品。 3．熟悉病例资料，了解患者的人口学特征和病情表现，根据患者的情况有所准备。 （1）人口学特征有研究表明青年人，尤其是男性青年依从性差；老年人可能会因记忆减退或身体上共病需要服多种药物而妨碍了依从性；女性比男性有较好的治疗依从性；文化层次越低，对疾病认识、治疗方案的理解有限，不清楚注意事项，依从性相对较差。 （2）病情表现相对而言，患者病情越重，症状越丰富，对病情的认识越差，依从性也越差；反之亦然。 4．查看近期各种治疗登记资料，分析患者的治疗依从性。 （1）找出患者不能顺利完成治疗的记录，登记拒绝参加和中途退出的次数、治疗种类，从而确定患者主要不依从治疗的种类及程度。 （2）对患者依从较好的治疗，登记治疗的种类。 5．护士做好以上准备工作后，向患者解释要对其治疗情况进行评估，希望患者配合。 6．征得患者同意后，安排一个安静不受打扰的环境进行评估。 7．访谈了解患者的经济状况和家庭社会支持情况。 （1）经济状况社会经济状况差、社会阶层低的患者依从性较差。经济状况好的患者依
电子商务简历中英文范文 个人简历最好用A4标准复印纸打印，字体最好采用常用的宋体或楷体，尽量不要用花里呼哨的艺术字体和彩色字体，排版要简洁明快，切忌标新立异，排的象广告一样。当然，如果你应聘的是排版工作则是例外。电子商务简历中英文范文篇一fwdq Female, 26, Education: college Working years: 3 to 5 years Expected salary: yuan Working location: chengdu to build Objective: electronic commerce competent specialist / | | shop operation content/new media operation promotion | | network shops operating Ability to perform Strong learning ability Have affinity Honesty integrity communication ability Work experience (work for 11 months, made a job) Star international health industry group Working time: March XX to February XX] [11 months Job title: general manager Education experience Graduated in July XX Sichuan chemical industry
眼镜行业目标市场分析 市场细分概念： 1：市场细分：在市场调查研究的基础上，依据消费者的需求、购买习惯和购买行为的异性，把整个市场细分成若干需要不同的产品和市场营销组合的子市场 （或细分市场）。 眼镜市场细分：在眼镜市场调查研究的基础上，依据汽车消费者的需求、购买习惯和购买行为的差异性，把整个汽车市场细分成若干需要不同的产品和市场营销组合的子市场（或细分市场）。 2：如何理解市场细分的概念？ 1）市场细分不是对产品进行分类，而是对消费者的需求和欲望进行分类。 2）每一个细分市场，都是一个由若干有相似的欲望和需求倾向的消费者构成的群体。 3）不同细分市场在需求倾向上的差异性，既表现在产品上，也表现在其他市场营销组合上。4）市场细分不是对市场简单分解，而是企业从更具体的角度寻找和选择市场机会。 3：为什么要进行市场细分？ 近几年来，中国眼镜产业年均增幅达17%。中国不仅已经成为世界潜力最大的眼镜消费大国，而且已成为世界领先的眼镜生产大国，眼镜产量已经占到企业界70%。眼镜行业效益水平良好，效益增幅高于产值增幅；从业人数持续增长。这些现象表明眼镜行业总体处于健康发展状态。 纵观近几年我国眼镜业的发展，从发展局面来看，我国眼镜行业形成了国有、集体、私营、合资、独资等多种所有制企业；从分布格局来看，中国眼镜企业主要分布在华东地，现已形成广东东莞、福建厦门、浙江温州、江苏丹阳、上海、北京等主要的生产基地，眼镜业出口主要集中在浙江温州、江苏丹阳和广东深圳，其中江苏丹阳已成为世界上最大的镜片生产基地和亚洲最大的眼镜产品集散地，镜架产量占国内三分之一，镜片产量占国内70%、占全球50%；总企业性质来看，外商和港澳台投资企业所占比重最大，其次是股份制企业。小型企业较多，大中型企业较少，且多为外商和港澳台投资企业或是股份制企业。目前，。在我国，对眼镜的需求呈现出极度多样化和个性化的特征 从品牌看，欧系、美系、日系、韩系，几乎世界所有国家的眼镜品牌和产品，在中国都有生存空间。 从眼镜类别看，不论是价值不菲的纯钛的材质的眼镜，还是价格一般的合金材质的眼镜，在中国都有市场。 从眼镜款式年代看，一边是越来越多的国际同步款型争相引入中国，一边是20年前的圆形黑框眼镜依然畅销。 因此，一个眼镜公司不可能生产满足所有消费者需求的眼镜，只能选择一部分消费者开展目标营销，这就需要对眼镜市场进行细分。 3.眼镜市场细分的好处 1）有利于企业发现新的最好的市场机会，从而选择适合本企业的目标市场。 2）有利于企业针对目标市场的需求特点，开发适销对路的产品，制定更有效的营销策略。3）有利于企业在产品定位、价格制定、广告策略和促销等营销要素的组合上正确的决策。4:市场细分的原则 可测量性可进入性可赢利性可区分性 5:市场细分的主要因素 心理因素环境因素人口因素行为因素 6:市场眼镜细分—整车市场 按外型分类：有全框、半框和隐型等
中英文求职信 提要：随信附上我的简历，如有机会与您面谈，我将十分感谢。即使贵公司认为我还不符合你们的条件，我也将一如既往地关注贵公司的发展，并在此致以最诚挚的祝愿。 更多内容通告 DearSir, Twoyearsasaseniorclerkin*salesdepartmentof*theGreatwallcompanyhave,Ibelievegivenmetheexpe riencetoqualifyforthejobyouadvertisedin51jobwebsite. Since2000Ihavebeenresponsiblefor*allofficedetailsintheadministrationofsales,,Ihavebecomefamili arwiththevarioussalesterritories,andhavealsoinmysparetimeexperienceofhandlingbusinessproblems otherthanmypropersphere. TheyearsbeforeIwasemployedattheGreatwall,Iwasasecretaryfor*LongDragon, Iwasgraduatedat*wuhanUniversity,*twenty-sevenyearsofageandsingle. Iamleavingmypresentpositio mayIseeyouatyourofficetotellyoumoreaboutmyselfandshowyoujusthowwellIcandotheworkyourequ ire. yoursfaithfully, 参考译文: 敬启者: 在51job人才网站上,获悉贵公司招聘人才。本人曾担任长城公司*销售部高级职员两年,自信有充分工作经验,可担任贵公司所招聘的职务。 本人自二零零零年开始,一直担任*售货业务以及函件的撰写工作。在工作期间,除正式业务范围外,与各地区客户颇有来往,并利用公余时间,学会了很多有关商业问题的处理。 在长城公司任职前,我曾在*长龙会计公司担任过秘书工作。在任职期间,学会很多有关会计名词及会计作业程序。 本人*现年二十七岁,未婚,一九九七年毕业于*武汉大学,我希望有机会充分利用自己工作能力,从事更大范围工作之需要,这是我急于离开现职的主要原国。本人现任职的公司老板,对我的工作雄心颇为赞许,因此,愿协助找另谋他职。 如果方便的话,我愿亲赴贵公司办公室晋见阁下,以便详细说明。
市场竞争状况分析计划书（模板） 一.市场背景分析 市场背景分析是非常重要的营销活动，也是开发区域市场迈出的第一步。只有通过周密的调研和分析，才能明确市场机会、市场威胁及自身的优劣势，从而为战略定位及营销策略提供决策依据。本将介绍与此相关的分析，内容涉及营销环境、消费者、竞争者、行业及企业自身共5个方面。 1.营销环境分析 1）人口统计 人口的一些相关资料因素如性别、年龄结构、教育水准、职业、家庭人数、地区人口数、总人口数、出生率、死亡率等，是用来区分购买者、进行市场细分的有用工具。 2）经济环境 市场不仅需要人口，而且还需要购买力。实际经济购买力取决于现行收入、价格、储蓄、负债，甚至信贷。 ①收入分配： 一般可以把收入分配分为5种类型：家庭收入极低；多数家庭低收入；家庭收入极低与家庭收入极高并存；低、中、高收入同时存在；大多数家庭属于中等收入。产品要寻找市场，必须在以上5种类型的分配结构中选择适宜的市场。 ②储蓄、债务、信贷的适用性： 营销人员必须注意收入、生活费、利息、储蓄和借款形式的变化，因为这对生产收入与价格敏感产品的企业特别具有重大影响。 3）法律法规环境 企业的定价、广告、促销等活动都将受到有关政策法规的限制，如专利法、商标法，商品检验法、关税法、消费者保护法、地方性法规等。 4）社会/文化环境
社会/文化反映着个人的基本信念、价值观和规范的变动，它会影响到企业的目标市场定位，营销活动必须符合社会文化的要求，才能顺应消费者的需求。 2.消费者状况分析 1）确定影响购买者购买行为的主要因素 ①文化因素： 文化因素对消费者的行为具有最广泛和最深远的影响。文化的层次分析: a．文化：如美国长大的儿童普遍有这样的价值观：成就与功名、活跃、效率与实践、上进心、物质享受、自我、自由、形式美、博爱和富有朝气。 b．亚文化：亚文化群体包括民族群体、宗教群体、种族团体和地理区域、社会阶层（如可以将社会阶层分为7个层次：上上层、上下层、中上层、中间层、劳动阶层、下上层、下下层。）。 ②社会因素： 影响消费者购买行为的社会因素有相关群体（包括家庭、朋友、邻居、同事等“主要群体”和、宗教、职业、贸易协会等“次要群体”）、家庭（包括“婚前家庭”和“子女家庭”，如丈夫支配型、妻子支配型、共同支配型）、和社会角色与地位。 ③个人因素： 包括年龄阶段、职业、经济环境、生活方式、个性等。 ④心理因素： 包括动机（需求）、知觉、学习及信念和态度。 2)五种类型的消费心理和模式 a．发烧型：是追求最新技术、最新产品的那一批人，但数量有限，他们对新发明新创造极感兴趣，愿意尝试不成熟的技术和产品，甚至自己动手参与个性或给厂家提出建议。在我国DVD和LD目前的消费群就属于这一类用户。 b．先锋型：是比较有远见、有追求的一批人，对技术、产品有一种敏感，愿意采用已
电子商务之诚信问题 一我国电子商务存在的诚信问题 中国电子商务诚信危机的表现 利用电子商务进行交易必然会涉及到信用与支付问题。由于电子商务的“无纸化”和“无址化”, 对参加交易的各方提出了更高的信用要求.处于转型期的中国社会,传统的“义理社会”价值体系的约束作用正在日趋削弱,而基于法制基础之上的“契约社会”还远未形成.信用的概念在不少人的眼中甚是淡薄,因此也给与电子商务密切配套的支付手段带来了很大的不利影响.目前国内所进行的电子商务交易,其支付手段可以说是土洋结合。信用卡、借记卡、储蓄卡、邮局汇款和货到付款等多种支付方式混合使用,有的甚至是使用网上查询、网下交易的方法.虽然现在有一些银行开始进行在线支付和开办网上银行业务等方面的试点工作,但是在中国信用制度还很不完善的情况下,单靠银行的力量也很难解决这一问题.而且,由于人与人间的信任度较差,很少有人愿意贸然通过网络的形式把自己的信用卡帐号等个人资料告诉企业,因为稍不留神就会发生想象不到的严重问题.现实中也确实发生过这样的情况,有人把信用卡帐号等个人资料在网上告诉了企业,被一些不良企业把不应收的货款划走,等到消费者发现已为时过晚。现在大多数从事电子商务的企业,都选择了货到付款这样一种较可靠的方式,以解决在货款的支付中双方互不信任的问题.但是,电子商务活动进行的最终目的,就是为了进行快捷、方便、安全的交易，使资金使用和货物流向趋于合理。如果我们仍然沿用传统的交易方法,使用现款支付的方式来实行网上交易，必然会制约企业电子商务的运作规模,而且违背了电子商务活动的初衷。 电子商务环节中存在的失信行为大体表现在一下几个方面： 1.货物品质失真 消费者利用电子商务网站网上购物，虽具有安全、方便等诸多的优点，但根据消费者反馈，通过上网所购的商品往往达不到消费者的心理预期，还存在以次充好的现象。例如作者经常在淘宝网上购物，往往在网站图片上看到的货品样式十分诱人，图片制作非常优美，可是等到购买之后，货物送到自己手上的时候才发现实物和网站上的图片相差甚，感觉非常不值。货品失真，是电子商务网上购物存在的一个非常严重的失信现象。 2．货物失踪 在网上购物的消费者投诉方面，消费者进行了网上支付，却没收到货物的投
中国眼镜行业市场现状和发展趋势分析 改革开放以来，我国眼镜行业所有制结构发生了很大变化，形成国有、集体、私营、合资、独资等多种所有制并存的局面，企业充满活力，眼镜行业从一个名不见经传的小行业，发展成一个充满生机，充满潜力的朝阳行业，我国也逐步发展成为世界主要眼镜消费国家和生产国家之一。目前中国不仅是世界潜力最大的眼镜消费大国，而且已成为世界领先的眼镜生产大国。 中国眼镜协会分析，近5年来，中国眼镜产业年均增幅达17％。目前，我国眼镜生产企业超过4000家，有一定规模的验光配镜店超过2万家，并已形成广东东莞、福建厦门、浙江温州、江苏丹阳、上海、北京等主要生产基地。眼镜的材质、品种、款式日益多元化，能够生产纯钛、钛合金、记忆金属等眼镜架、角膜接触镜、CR-39光学树脂镜片、镀膜镜片、渐进多焦点镜片、非球面镜片等高技术含量的新型眼镜产品，并出现江苏“万新”、浙江“海豚”等知名品牌。据保守估计，中国约有3亿人配戴眼镜（矫正屈光镜），按每3年更新一付来计算，每年的市场需求量就达1亿付，如果加上太阳镜，市场需求量就更为可观。除了实用功能，眼镜还被赋予装饰功能，眼镜消费的个性化、时尚化、品牌化、高档化的趋势日益显着，这进一步催生了中国眼镜市场的商机。 受中国市场诱惑，几乎所有的国际知名眼镜厂家和商家纷纷来华掘金，或设立生产基地，或编织销售网络。譬如，1997年，全球最大的镜片制造商法国依视路(ESSILOR)投资5300万美元在上海建立了镜片生产基地，产量每年以两位数的速度增长，并占据了12％的中国市场。日本野尻1989年登陆中国，不仅在上海设立了镜架生产厂，而且在全国布局了上千家连锁店。香港是全球第二大眼镜生产中心，
求职信中英文对照范文 求职信是我们求职过程中的一个重要环节，你知道你的中文求职信怎么翻译成英文吗?下面由为你提供的求职信中英文对照范文，希望能帮到你。 求职信中英文对照范文(一)尊敬的先生/小姐： 您好!我从报纸上看到贵公司的招聘信息，我对网页兼职编辑一职很感兴趣。 我现在是出版社的在职编辑，从1998年获得硕士学位后至今，一直在出版社担任编辑工作。两年以来，对出版社编辑的工作已经有了相当的了解和熟悉。经过出版者工作协会的正规培训和两年的工作经验，我相信我有能力担当贵公司所要求的网页编辑任务。 我对计算机有着非常浓厚的兴趣。我能熟练使用FrontPage和DreamWeaver、PhoteShop等网页制作工具。本人自己做了一个个人主页，日访问量已经达到了100人左右。通过互联网，我不仅学到了很多在日常生活中学不到的东西，而且坐在电脑前轻点鼠标就能尽晓天下事的快乐更是别的任何活动所不及的。 由于编辑业务的性质，决定了我拥有灵活的工作时间安排和方便的办公条件，这一切也在客观上为我的兼职编辑的工作提供了必要的帮助。基于对互联网和编辑事务的精通和喜好，以及我自身的客观
条件和贵公司的要求，我相信贵公司能给我提供施展才能的另一片天空，而且我也相信我的努力能让贵公司的事业更上一层楼。 随信附上我的简历，如有机会与您面谈，我将十分感谢。即使贵公司认为我还不符合你们的条件，我也将一如既往地关注贵公司的发展，并在此致以最诚挚的祝愿。 此致 求职信人：xxx 20xx年xx月xx日 April 13,2000 P.O. Box 36 BIIT University Beijing,China 100000 Dear Sir/Madam: Are you searching for a telecommunications manager with expertise in project management and team leadership? I specialize in creating and implementing high-performance strategies that directly impact growth and profitability of large telecommunications companies. In addition to my knowledge of business processes, I also offer proficiency in telecom software development and cutting-edge technologies. I am relocating to Shenzhen and would be interested in
住院患者服药依从性管理及影响因素分析 目的了解住院患者服药的依从性及相关影响因素。方法选择在我院住院治疗的患者作为研究对象，依照Morisky等推荐的用药依从性标准对患者用药依从性进行评定，采用χ2检验和Logistic回归分析筛选患者用药依从性的影响因素。结果157例患者中，服药依从者96例，依从率61.1%；服药不依从者61例，不依从率38.9%。多因素Logistic回归分析显示药费支付方式、用药时间、是否需要长期服药的知晓情况对住院患者服药依从性有影响。表现为：公费或医保患者依从性高于自费患者（OR=1.248）；服药时间在2年～和≥3年患者的依从性高于服药时间<3个月患者（OR=1.240和2.597）；需要长期服药患者的依从性比不需要长期服药者更高（OR=2.213）。结论应加强用药患者的依从性教育，努力提高患者的用药依从性。 标签：住院患者；药物治疗；依从性；影响因素 住院患者往往都要经历漫长的疾病恢复期，有很多患者需要长期服药甚至终身服药，患者服药依从性无论对于在家服药患者还是住院患者都是一个普遍存在的问题，对住院患者进行科学、系统的用药指导，培养其良好的服药习惯、提高患者的服药依从性，对于提高患者治疗效果、减低并发症等都有重要意义[1]。为了了解我院住院患者服药依从性程度及其影响因素，以便為患者设计更好的治疗方案，更好的指导患者用药，最终达到提高治疗效果的目的[2-3]。为此，我们以问卷调查的形式对四川省仁寿县中鑫医院的157例住院患者进行了服药依从性的临床调查研究。 1 资料与方法 1.1一般资料选择2014年4月～12月在四川省仁寿县中鑫医院住院治疗的住院患者160例，作为研究对象，发放调查问卷160份，最终收回有效问卷157份，有效回收率98.13%。 1.2方法服药依从性判定标准：依照Morisky等推荐的服药依从性标准判定患者的服药依从性，依从性量表：①你是否有忘记点药的经历；②你是否为了快速达到目的，而自己增加服药次数；③当你自觉症状改善时，是否会自行停药； ④当你觉得药效不明显时，是否会自行停药。当有≥3个条目回答为”否”则判定为依从，有≤2个条目回答为”否”则判定为不依从。同时对患者的基本情况、疾病情况、治疗情况等进行问卷调查。 1.3统计学处理数据资料通过Epidata3.10进行管理，采用SPSS16.0统计软件对数据资料进行统计学分析，检验水准α为0.05。单因素分析采用χ2检验对各调查因素的服药依从性进行分析，采用多因素Logistic回归分析筛选影响患者服药依从性的影响因素。 2 结果
天津大学《电子商务》课程教学大纲 课程编号：2160259 课程名称：电子商务1 学 时： 32 学 分： 1.5 学时分配： 授课：16 上机：16 实验： 实践： 实践（周）： 授课学院： 计算机科学与技术学院 适用专业： 计算机科学与技术 先修课程： 程序设计原理，计算机网络，Web开发技术 一．课程的性质与目的 电子商务课程是一门用以培养学生了解电子商务原理，设计电子商务网站能力的技术选修课。教学过程中综合运用先修课程中所学到的有关知识与技能，结合各种实践教学环节，进行IT工程技术人员所需的基本训练，为学生进一步学习有关专业课程和日后从事电子商务设计工作打下基础。 二．教学基本要求 1、掌握设计电子商务网站的基本理论、基本知识和基本技能，了解电子商务 网站的架构并进行相应设计的能力； 2、具有综合运用各种工具设计电子商务网站工具的能力； 3、掌握电子商务应用的部署能力； 4、了解电子商务的新理论、新方法及发展趋向。 三．教学内容 1、电子商务概述 电子商务的定义与分类 电子商务的发展现状 随需应变（On-demand）电子商务 2、构建Web应用技术 客户/服务器模型，COOKIE和SESSION的运用 WEB应用模型，三层和n层架构模型 Web Service的体系架构和SOAP，UDDI，WSDL
SOA的体系架构，SOA与Web服务的关系，ESB，工作流 3、盈利模式 Web目录盈利模式 数字内容盈利模式 广告支持的盈利模式 广告-订阅混合式盈利模式 按交易付费式盈利模式 按服务付费式盈利模式 4、电子商务模式 商业模式 集成模式 复合模式 定制设计 应用程序模式 运行时模式 5、电子商务安全技术 电子商务安全的基本概念 加密与验证，防火墙技术，数字证书 身份管理，单点登录 Web Service安全 6、电子支付技术 电子支付概述 安全套接层协议SSL 安全电子交易SET协议 电子支付工具 网上银行 7、随需应变解决方案的设计 方案设计师的角色 方案设计流程