Paeonol from Hippocampus kuda Bleeler suppressed the neuro-in?ammatory responses in vitro via NF-j B and MAPK signaling pathways
S.W.A.Himaya a ,BoMi Ryu b ,Zhong-Ji Qian b ,c ,Se-Kwon Kim a ,b ,?
a
Biochemistry and Molecular Biology Laboratory,Department of Chemistry,Pukyong National University,Busan 608-737,Republic of Korea b
Marine Bioprocess Research Center,Pukyong National University,Busan 608-737,Republic of Korea c
Department of Marine Life Science and Marine Life Research &Education Center,Chosun University,Gwangju 501-759,Republic of Korea
a r t i c l e i n f o Article history:
Received 19April 2011Accepted 11April 2012
Available online 27April 2012Keywords:Paeonol Microglia
Anti-in?ammation MAPK NF-j B
a b s t r a c t
In?ammation has recently been implicated as a critical mechanism responsible for neurodegenerative diseases.In this study,paeonol (1-(2-hydroxy-4-methoxyphenyl)ethanone)isolated from the sea horse Hippocampus kuda Bleeler was studied as an agent to suppress LPS induced activation of BV-2microglial and RAW264.7macrophage cells.The results obtained showed that paeonol signi?cantly suppressed LPS induced release of pro-in?ammatory products such as nitric oxide (NO),prostaglandin E2(PGE 2),and cytokines;tumor necrosis factor-a (TNF-a ),interleukin-1b (IL-1b )and interleukin-6(IL-6).Furthermore,the compound down regulated the protein and gene expression levels of inducible nitric oxide synthase (iNOS),cyclooxygenase-2(COX-2),TNF-a ,IL-1b and IL-6in both cell lines.Molecular signaling pathway studies showed that paeonol inhibited the translocation of nuclear factor-j B (NF-j B)p65and p50sub-units to the nucleus by blocking IKK a /b (I j B kinase a /b )mediated degradation of I j B a .Moreover,it suppressed the phosphorylation of mitogen activated protein kinase (MAPK)pathway molecules;c-Jun N-terminal kinases (JNK)and p38in both cell lines.Collectively these results indicate that paeonol blocked the LPS stimulated in?ammatory responses in BV-2and RAW264.7cells via modulating MAPK and NF-j B signaling pathways.Therefore,paeonol could be a promising candidate to be used in neuro-in?ammatory therapy.
ó2012Elsevier Ltd.All rights reserved.
1.Introduction
In?ammation has been implicated as the pathophysiological mechanism underlying many chronic diseases,including diabetes,cardiovascular disease,cancer,arthritis,and neurodegenerative diseases (Lawrence et al.,2002;Libby,2007).The involvement of in?ammation as a potential pathogenic factor in chronic neurode-generative diseases such as Alzheimer’s disease,Parkinson’s dis-ease,HIV dementia and multiple sclerosis has been studied extensively in the recent past (Ekdahl et al.,2009).Central nerves system (CNS)resident mono-nuclear phagocyte populations;microglia and macrophages play a central role in initiating the
in?ammatory response (Block et al.,2007;Perry et al.,2007).At an incidence of systemic in?ammatory signaling these cells are readily activated and initiate the secretion of neurotoxic cytokines including tumor necrosis factor-a (TNF-a ),interleukin 1b (IL-1b ),interleukin-6(IL-6)reactive oxygen species (ROS)and prostaglan-dins.These molecules are associated in the subsequent progression of the neuron damage.
The in?ammatory responses in the microglial cells and macro-phages are found to be mainly regulated by NF-j B (Ock et al.,2009).NF-j B is a transcription factor that mediates immune and in?ammatory responses.The activation of NF-j B involves phos-phorylation and degradation of I j B bound to NF-j B,via signaling through IKK phosphorylation.The resulting free NF-j B is translo-cated into the nucleus to promote the expression of pro-in?amma-tory mediators such as iNOS,COX-2and cytokines (Cheong et al.,2011).Further,MAPK signaling pathways (JNK,ERK and p38)are also involved in modulation of in?ammatory responses through up-regulating the expression of cytokines (Gao et al.,2012).
Chinese seahorse (Hippocampus kuda Bleeler)is one of the most famous material in traditional Chinese medicine and has been studied for many years for its various biologic activities,including appetite enhancement,antioxidant,anti-tumor,anti-aging and
0887-2333/$-see front matter ó2012Elsevier Ltd.All rights reserved.https://www.doczj.com/doc/bb279323.html,/10.1016/j.tiv.2012.04.022
Abbreviations:CNS,central nervous system;MTT,3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide;LPS,lipopolysaccharide;PGE 2,prostaglandin E 2;iNOS,inducible nitric oxide synthase;COX-2,cycloxygenase-2;TNF-a ,tumor necrosis factor-a ;IL-1b ,interleukine-1b ;IL-6,interleukine 6;MAPK,mitogen-activated protein kinase;JNK,Jun N-terminal kinase;ERK,extracellular signal-regulated kinase;IKK a /b ,I j B kinase a /b ;NF-j B,nuclear factor-j B.
?Corresponding author at:Biochemistry and Molecular Biology Laboratory,Department of Chemistry,Pukyong National University,Busan 608-737,Republic of Korea.Tel.:+82516297094;fax:+82516297099.
E-mail address:sknkim@pknu.ac.kr (S.-K.Kim).
anti-fatigue activities(Li et al.,2008;Ryu et al.,2010;Zhang et al., 2003).As a part of an ongoing research on isolating bioactive metabolites from sea horse H.kuda Bleeler,the compound paeonol [1-(2-hydroxy-4-methoxyphenyl)ethanone]was isolated.Previ-ously,it has been found that paeonol is an active component in; moutan cortex(Paeonia suffruticosa Andrews,Ranunculaceae),a Chinese medicine(Lau et al.,2007;Li et al.,2009;Wu and Gu, 2009),roots of https://www.doczj.com/doc/bb279323.html,cti?ora Pallas,a traditional Chinese herb(Niza-mutdinova et al.,2007)and a Chinese yam,Dioscorea japonica (Miyazawa et al.,1996).Furthermore,paeonol has been character-ized for its inhibitory activity on TNF-a and IL-6production,anti-oxidant effect,anti-atherosclerosis effect,anti-diabetic effect and anti-mutagenic effect.However,no studies have been reported on the anti-neuroin?ammatory activity of paeonol up to date. Therefore,this study was conducted with the aim of investigating paeonol as a candidate natural metabolite to regulate chronic neu-ro-in?ammatory responses in LPS activated human microglial cells (BV-2)and murine macrophages cells(RAW264.7).Furthermore, efforts have been taken to identify the signaling pathways under-lying the anti-in?ammatory effect of paeonol.
2.Materials and methods
2.1.Materials and chemicals
Following instruments and chemicals have been used in this study to perform the experiments.1H NMR(400MHz)and13C NMR(100MHz)spectra were recorded on a JEOL JNM-ECP400 NMR spectrometer(JEOL,Japan),using CDCl3(1D,12.75ppm in 1H and113–9ppm in13C NMR)as the solvent.MS spectra were ob-tained on a JEOL JMS-700spectrometer(JEOL,Japan).Silica gel60 (230–400mesh,Merck,Germany)was used as the resin when per-forming column chromatography.Thin-layer chromatography (TLC)plates(Kieselgel60F254,0.25mm,Merck)were used in ana-lytical TLC.
Mouse microglial cells were a kind gift by professor Il-Whan Choi from Inje University,Korea.Mouse macrophages,RAW264.7cells were obtained from the American type culture collection(Manas-sas,VA,USA).Cell culture media[Dulbecco’s modi?ed Eagle’s min-imal essential medium(DMEM)],penicillin/streptomycin,fetal bovine serum(FBS)and the other materials required for culturing cells were purchased from Gibco BRL,Life Technologies(Grand Is-land,NY).The MTT reagent[(3-(4,5-dimethyl-2-yl)-2,5-diphenyl-tetrazolium bromide)],the Griess reagent and LPS of Escherichia coli026:B6were purchased from Sigma Chemicals.Primary and sec-ondary antibodies used for Western blot analysis were purchased from Santa Cruz Biotechnology Inc.(Santa Cruz,CA)and Amersham Pharmacia Biosciences(Piscataway,NJ).Other chemicals and re-agents used were of analytical grade available commercially.
2.2.Extraction and isolation
The live adults of sea horse were collected from Zhoushan Island, Zhejiang,China,in October,2005,and freeze-dried after removing the internal organs.The species was identi?ed as H.Kuda Bleeler by Professor Ming-Lu Deng(zoologist,Changchun University of Chi-nese Medicine,China).The freeze-dried H.Kuda Bleeler samples (3kg)were ground into powder and re?uxed with MeOH (3?5L)for3h,and the solvent was evaporated in vacuo to obtain the crude MeOH extract(217g).The extract was subjected to silica gel?ash chromatography by eluting with n-hexane/EtOAc/MeOH (gradient),and afforded10main fractions.Fraction6(502mg, EtOAc)was puri?ed over Silica gel repeatedly.The successful isola-tion resulted in36.05mg of the puri?ed https://www.doczj.com/doc/bb279323.html,prehensive spectral analysis was carried out to identify the structure of the compound,including LREIMS(low resolution electron impact mass spectra),1H NMR(nuclear magnetic resonance),13C NMR,and13C DEPT(distortionless enhancement by polarization transfer).
2.3.Culture of BV-2and RAW264.7cells
BV-2and RAW264.7cells were cultured in Dulbecco’s modi?ed Eagle’s medium(DMEM)supplemented with5%heat-inactivated fetal bovine serum(FBS),100l g/ml streptomycin,and100l g/ml penicillin in a humidi?ed atmosphere of5%CO2and95%O2at 37°C.RAW264.7cells and BV-2cells were maintained via two times passage per week and cells were utilized for experimenta-tion at70–80%con?uency.
2.4.Nitric oxide production assay
Nitric oxide(NO)levels in the culture supernatants were mea-sured by the Griess reaction as described in previously(Coker and Laurent,1998).In brief,BV-2and RAW264.7cells were pre-incubated for24h in96-well plates using DMEM without phenol red at a density of1?104cells per well,followed by the treatment of paeonol(5,10and50l M)for2h.Then the NO production was stimulated by incubating with LPS(1l g/ml?nal concentration)for 48h.Fifty microliter of culture supernatants from each sample was collected and mixed with a same volume of Griess reagent[1%sul-fanilamide/0.1%N-(1-naphthyl)-ethylenediamine dihydrochloride/ 2.5%phosphoric acid]and allowed to incubate for15min.Absor-bance values were read at540nm on an ELISA microplate reader (Tecan Austria GmbH).Nitrite concentration in the conditioned media was calculated with reference to a standard curve of sodium nitrite generated by known concentrations.
2.5.Enzyme immuno assay of PGE2
Assessment of PGE2synthesis was performed by enzyme immu-noassay using commercially available PGE2enzyme immunomet-ric assay kit(Amersham Pharmacia Biosciences,NJ,USA).BV-2 and RAW264.7cells were treated with different concentrations of paeonol(5,10and50l M)for2h and stimulated with LPS(1l g/ ml)for24h the conditioned media was collected and analyzed according to the manufacturer instructions.The concentration of PGE2was calculated with reference to the standard curve obtained by PGE2standard solution provided with the EIA kit.
2.6.Enzyme immuno assay of TNF-a,IL-1b and IL-6
The inhibitory effects of paeonol on the production of pro-in?ammatory cytokines;TNF-a,IL-1b and IL-6were determined by an enzyme-linked immunosorbent assay(ELISA).Cells were treated with different concentrations of paeonol(5,10and 50l M)for2h followed by LPS(1l g/ml)treatment for24h.Col-lected conditioned media were analyzed per the manufacturer’s recommendations of mouse cytokine-speci?c Biotral?ELISA kits (Amersham Pharmacia Biosciences,NJ,USA).The concentrations of TNF-a,IL-1b and IL-6were calculated according to the standard curves generated by each of the recombinant cytokines provided with the ELISA kits.
2.7.Reverse transcription polymerase chain reaction(RT-PCR)analysis
RT-PCR was performed to detect the mRNA expression levels of iNOS,COX-2,and cytokines(TNF-a,IL-1b and IL-6).Total RNA was extracted from BV-2and RAW264.7cells after treatment with paeonol at different concentrations(5,10and50l M)for2h followed by LPS(1l g/ml)treatment for18h.The cells were lysed with Trizolòand centrifuged at12,000rpm for15min at25°C
S.W.A.Himaya et al./Toxicology in Vitro26(2012)878–887879
following the addition of chloroform(5:1).Supernent was separated and isopropanol was added to it at a1:1ratio.By centri-fugation for10min at10,000rpm,RNA pellet was obtained.After washing with ethanol,extracted RNA was solubilized in diethyl pyrocarbonate(DEPC)treated RNase-free water and RNA amount was quanti?ed by measuring the absorbance at260nm using the GENiosòmicroplate reader(Tecan Austria GmbH).Resulted RNA (2l g)were reverse transcribed into cDNA using a master-mix con-taining1?reverse transcriptase(RT)buffer,1mM dNTPs,500ng of oligo(dT)15primers,140U of murine Moloney leukemia virus (MMLV)reverse transcriptase and40U of RNase inhibitor,for 45min at42°C.Polymerase chain reaction(PCR)was carried out in an automatic Whatman thermocycler(Biometra,Kent,UK)to amplify iNOS,COX-2,TNF-a,IL-1b,IL-6and glyceraldehyde-3-phosphate dehydrogenase(GAPDH)mRNA as the gene control. Each transcript was identi?ed using speci?c forward and reverse primers as manufacturer’s instructions(Promega,Madison,WI, USA).The conditions for the ampli?cation cycles were95°C for 30s,55°C for30s,and72°C for1min for35cycles.After poly-merase chain reaction products were electrophoresed on1.5%aga-rose gels in1?TAE buffer for30min at100V and were visualized by ethidium bromide staining using AlphaEaseògel image-analysis software(Alpha Innotech,San Leandro,CA,USA).Primer sequences used to amplify the desired cDNA were as follows:iNOS forward and reverse primers:50-CCCTTCCGAAGTTTCTGGCAGCAGC-30 and50-GGCTGTCAGAGCCT CGTGGCTTTGG-30;COX-2forward and reverse primers:50-GGGGTACCTTC CAGCTGTCAAAATCTC-30and 50-GAAGATCTCGCCAGGTACTCACCTG-30;TNF-a forward and re-verse primers:50-ATGAGCACAGAAAGCATGATC-30and50-TACA GGCTTGTCACTCGAATT-30;IL-1b forward and reverse primers: 50-ATGGCAACTGTTCCTGAACTCAACT-30and50-TTTCCTTTCTTAGA TATGGACAGGAC-30;IL-6forward and reverse primers:50-AGTTGCCTTCTTGGGACTGA-30and50-CAGAATTGCCATTGCACAAC-30; and G3PDH forward and reverse primers:50-TGAAGGTCGGTGT GAACGGATTTGGC-30and50-CATGTAGGCCATGAGGTCCACCAC-30.
2.8.Western blot analysis
Standard procedures were used for the Western blotting,in brief;BV-2and RAW264.7cells treated with different concentra-tions of paeonol(5,10and50l M)followed by LPS treatment (1l g/ml),were lysed in lysis buffer containing50mM Tris–HCl (pH7.5),0.4%Nonidet P-40,120mM NaCl,1.5mM MgCl2,2mM phenylmethylsulfonyl?uoride,80l g/ml of leupeptin,3mM NaF and1mM DTT at4°C for30min.Remaining cell debris were re-moved by centrifugation.
To obtain the nuclear extracts of BV-2and RAW264.7cells,Cel-Lytic?NuCLEAR?extraction kit(Sigma–Aldrich Co.,Mo,USA)was used according to the manufacturer’s instructions.The protein concentrations of the cell lysates were determined using Lowry method.Cell lysates contain equal amount of proteins(%20l g of total protein)were separated by SDS–PAGE gel electrophoresis and electro-blotted onto a nitrocellulose membrane.The mem-branes were blocked with5%BSA and then incubated with desired primary and secondary antibodies(Santa Cruz Biotechnology Inc.). The protein expressions were detected by chemiluminescent ECL assay kit(Amersham Pharmacia Biosciences),according to the manufacturer’s instructions.Blots were visualized using an LAS3000òLuminescent image analyzer and protein expression lev-els were quanti?ed by Multi Gauge V3.0software(Fuji?lm Life Sci-ence,Tokyo,Japan).
2.9.Statistical analysis
All the data were presented as mean±S.D from three indepen-dent experiments unless stated otherwise and the error bars represent the standard deviation.Biologic variability between re-sults were identi?ed through statistical comparisons between dif-ferent treatments using one way ANOVA employing Student Newman Keul’s post hoc test in SPSS program(version12.0).The sig-ni?cance level of P<0.05was considered to be statistically signi?cant.
3.Results
3.1.Elucidation of paeonol
On the basis of the comprehensive spectral analysis and data pub-lished previously,the isolated compound was elucidated as a known 1-(2-hydroxy-4-methoxyphenyl)ethanone(Paeonol)(Fig.1).The molecular formula of paeonol was determined as C9H10O3.
Paeonol:1H NMR(400MHz,CDCl3):d H ppm12.75(1H,s,HO-2), 7.62(1H,d,J=8.7Hz,H-6),6.44(1H,dd,J=8.7,2.4Hz,H-5),6.42 (1H,d,J=2.4Hz,H-3),3.84(3H,s,H-9),2.55(3H,s,H-8);13C NMR (100MHz,CDCl3):d c ppm113.9(C-1),165.2(C-2),100.8(C-3), 166.1(C-4),107.6(C-5),132.3(C-6),202.6(C-7),26.2(C-8),55.5 (C-9)(see Table1);LREIMS m/z166.05(76)[M]+,151.05(100) [M-CH3]+,123.05(4)[M-CH3–CO]+,108.00(23)[M-CH3–CO–CH3]+,95.05(31),79.95(10),51.00(17).
3.2.Paeonol inhibited the LPS induced NO production via suppressing iNOS expression
In this study,nontoxic concentrations of paeonol(5,10and 50l g/ml)was tested for its ability to inhibit the LPS stimulated NO production(data on toxicity assessment is not shown).Nitrite
of the1-(2-hydroxy-4-methoxyphenyl)ethanone
Table1
NMR data for paeonol a.
Position Paeonol
d H(mult,J)d C(mult)
1113.9(d) 2165.2(s)
3 6.42(1H,d,2.4Hz)100.8(d)
4166.1(s)
5 6.44(1H,dd,2.4,8.7Hz)107.6(d)
67.62(1H,d,8.7Hz)132.3(d)
7202.6(s)
8 2.55(3H,s)26.2(q)
9 3.84(3H,s)55.5(q)
10
11
2-OH12.75(1H,s)
a Recorded in CDCl
3
at400Hz for1H and100Hz for13C.
880S.W.A.Himaya et al./Toxicology in Vitro26(2012)878–887
2.Effects of paeonol on LPS induced NO production in BV-2(A)and RAW264.7(B)cells.Cells cultured in phenol red and serum free media were pre-treated with different concentrations(5,10and50l M)of paeonol for1h,followed by stimulation with LPS(1l g/ml).Conditioned medium was collected after48h and NO concentrations were measured via the Griess reaction.The nitrite(stable oxidation product of NO)levels in the conditioned media were calculated compared to standard nitrite curve.The indicate the mean±SD of three independent experiments.Paeonol inhibited LPS-induced iNOS protein and mRNA expressions in BV-2(C)and RAW264.7(D)cells.Cells were pretreated with paeonol(5,10and50l M)for1h,and then stimulated with LPS(1l g/ml)for18h.Cell lysates were extracted,and protein and gene levels of iNOS were analyzed by Western blotting and RT-PCR respectively.b-Actin and GAPDH was used as an internal controls.The expression levels of iNOS protein and gene were quanti?ed
a percentage compared to the LPS alone treated control group.The data were con?rmed by three independent experiments.?P<0.05indicates sample treatments were signi?cantly different from the value in LPS alone treated control group at the given signi?cance level.
3.Effects of paeonol on LPS induced PGE2production in BV-2(A)and RAW26
4.7(B)cells.Cells cultured in phenol red and serum free media were pre-treated with different concentrations(5,10and50l M)of paeonol for1h,followed by the stimulation with LPS(1l g/ml).Conditioned culture media was collected after24h in order measure PGE2concentrations by the enzyme immunoassay.The data indicate the mean±SD of three independent experiments.?P<0.05and#P<0.01compared to control. Paeonol inhibited LPS-induced COX-2protein and mRNA expressions in BV-2(C)and RAW264.7(D)cells.Cells were pretreated with paeonol(5,10and50l M)for1h, then stimulated with LPS(1l g/ml)for18h.Cell lysates were extracted,and protein and gene levels of iNOS were analyzed by Western blotting and RT-PCR respectively. Actin and GAPDH expressions were used as an internal controls.The expression levels of COX-2protein and gene were quanti?ed as a percentage compared to the LPS alone treated control group.The data were con?rmed by three independent experiments.?P<0.05,#P<0.01indicates sample treatments were signi?cantly different from the value LPS alone treated control group at given signi?cance level.
protein and mRNA expressions obseved after treatment with paeo-nol in LPS stimulated BV-2and RAW264.7cells,respectively.It could be clearly see that both protein and mRNA expressions were inhib-ited by the treatment of paeonol in both cell lines.This is in agree-ment with the results of paeonol attenuated decrease in PGE 2production.Therefore it could be suggested that paeonol induced inhibition of PGE 2production would due to inhibition of its up-stream enzyme COX-2at its transcriptional and translational levels.
3.4.Paeonol inhibited the LPS induced pro-in?ammatory cytokines production and expression
Pro-in?ammatory cytokines are key components of in?amma-tion.In the CNS,proin?ammatory cytokines produced from activated microglia or in?ltered macrophages is involved in patho-genesis of brain in?ammation.Therefore,the effect of paeonol on the production levels of in?ammatory cytokines TNF-a ,IL-1b
and
inhibited the production of LPS-induced TNF-a (A and B),IL-1b (E and F)and IL-6(I and J)in BV-2and RAW264.7cells.cells were pretreated h,and then stimulated with LPS (1l g/ml).Culture media were collected after 24h of treatment in order to measure the cytokine indicate the mean ±S.E.M of three independent experiments.Paeonol inhibited the gene expression of LPS-induced TNF-a (C and D),IL-1pretreated with paeonol for 1h,following LPS stimulation for 18h.Total RNAs were isolated,and mRNA levels of TNF-a were was used as an internal control.The data were con?rmed by three independent experiments.?P <0.05,#P <0.01indicates sample different from the value in LPS alone treated control group at given signi?cance level.
IL-6were investigated by ELISA using conditioned media of LPS challanged BV-2and RAW264.7cells.LPS stimulation have signif-icantly increased the production of pro-in?ammatory cytokines compaired to non-stimulated blank groups.LPS induced produc-tion of TNF-a,IL-1b and IL-6showed a signi?cant(P<0.05)con-centration dependent decrease(Fig.4A,B,E,F,I and J)following the treatment of paeonol(10,50and100l g/ml).In addtion,the transcriptional levels of TNF-a,IL-1b and IL-6were analyzed by RT-PCR using the RNA isolated from paeonol treated LPS chal-lenged cells.The results obtained were parallel with the above data signi?cant in both cell lines.These results indicates that paeonol mediated inhibition of pro-in?ammatory mediators and cytokines was probably occured via inhibition of MAP kinases signal trans-duction pathways;JNK and p38.
3.6.Paeonol blocked the NF-j B activation and translocation
NF-j B pathway plays a seminal role in in?ammation related pathogenesis,as it activates pro-in?ammatory genes encoding
5.Paeonol suppressed the LPS induced phosphorylation of MAPK pathway molecules in BV-2(A)and RAW264.7(B)cells.Protein expression levels of phosphorylated forms of three MAPK molecules,ERK,JNK and p38MAPK was assessed by Western blotting.To analyze phosphorylation levels of MAPK molecules cellular extracts were prepared from BV-2and RAW264.7cells pretreated with paeonol(5,10and50l g/ml)for1h,followed by stimulated with LPS(1l g/ml)for30min.The protein expression levels were quanti?ed as a percentage compared to the LPS alone treated control group.The data were con?rmed by three independent experiments.?P<0.05indicates sample treatments were signi?cantly different from the value in LPS alone treated control group at given signi?cance level.
884S.W.A.Himaya et al./Toxicology in Vitro26(2012)878–887
level.
by the treatment with paeonol in both BV-2and RAW264.7cell lines.This I j B a degradation has decreased through paeonol medi-ated inhibition of IKK a/b(Fig.6A and B),which induces the degra-dation of I j B a.To?nd the effect of paeonol on nuclear translocation of NF-j B;protein levels of NF-j Bp65and NF-j Bp50in the nucleus was analyzed.In Fig.6C and D it was clearly shown that LPS treatment induces the nuclear translocation of NF-j Bp65and NF-j Bp50subunits when compared to non LPS challenged blank group in both cell lines.And this translocation of NF-j Bp65and NF-j Bp50subunits were dose dependently inhibited by the treatment of paeonol in both cell types.These re-sults suggested paeonol suppress the signaling cascade of NF-j B pathway and nuclear translocation of NF-j B in LPS induced BV-2 and RAW264.7cell lines.
4.Discussion
Activated microglial cells and macrophages are one of the most important types of effector cells in mediating in?ammatory re-sponses in the central nerve system(CNS).Chronic activation of these cells due to internal or external factors causes damage to neurons due to unregulated release of neuro-toxic factors(Pan et al.,2009).This phenomenon is identi?ed as one of the major underlying cause of neuron cell death during neuro-degenerative diseases such as Alzhehimer’s disease,Parkinson’s disease,Dimetia and multiple sclerosis.Effective use of anti-in?ammatory agents to combat the neuro-degenerative diseases has gained much atten-tion recently.Natural compounds isolated from marine organisms have shown potent anti-in?ammatory activity due to their unique structural features.In the present study paeonol isolated from Chi-nese sea horse was analyzed for its capacity to suppress the neuro-toxic factors and their upstream molecule targets produced during the pathogenesis of neuroin?ammation.Paeonol is a water soluble low molecular weight(166.05)compound with active functional groups such as methoxy,hydroxyl and ketone groups attached to it.The functional groups have the ability to scavenge neurotoxic radical species by donating electrons(Flora,2009).Interestingly, scienti?c evidences con?rms that the compounds bearing methoxy (àOCH3)groups are of high anti-in?ammatory activity(Srinivasan et al.,2009).Collectively these structural features of paeonol sup-ports the shown anti-in?ammatory activity of paeonol.
In experimental models activation of microglial and macro-phage cells could be mimicked by LPS treatment.LPS is a bacterial lipopolysaccharide which has the potency to trigger in?ammatory responses in microglial and macrophage cells(Pan et al.,2009).LPS stimulation induce expression of iNOS which triggers the produc-tion of its downstream effector molecule nitric oxide(NO)which ultimately results in neuronal damage by NO itself or its more-toxic metabolite,peroxynitrite(ONOOà)(Dringen,2005;Li et al., 2005).In the similar manner the inducible cyclooxygenase,COX-2drives the onset of in?ammation through the production of pro-in?ammatory prostaglandin E2(Tzeng et al.,2005).Accumu-lating evidences have shown that COX-2plays a crucial role in pathogenesis of brain in?ammation(Scali et al.,2003).Pro in?am-matory cytokines;TNF-a,IL-1b and IL-6are largely produced in an in?ammatory condition and these cytokines act as messengers which stimulate the in?ammatory process(Jung et al.,2009a). Therefore,down regulation of these pro-in?ammatory cytokine production is of utmost importance during anti-in?ammatory therapy.As demonstrated in the results paeonol have effectively suppressed these cytokine productions via inhibiting their respec-tive gene expressions.
In?ammatory process is mainly regulated by NF-j B signaling pathway in both microglial and macrophage cells.NF-j B is activated by phosphorylation,ubiquitination,and subsequent proteolytic degradation of the NF-j B bound protein I j B via acti-
vated I j B kinase(IKK)(Rajapakse et al.,2008).The liberated NF-j B transcription factor translocates to the nucleus and binds to
j B motifs in the promoters of target genes such as iNOS,COX-2 and cytokines,to promote their transcription(Lee et al.,2003). Paeonol has suppressed the NF-j B activation and thereby inhibit-ing the transcription of in?ammatory genes.
MAPK signaling pathways are also directly involved in the syn-thesis of proin?ammatory mediators in activated macrophages (Jung et al.,2009b)and microglia(Pan et al.,2009).Aberrant phos-phorylation of MAPKs molecules are signaling to the expression of pro-in?ammatory mediators.LPS induce a rapid phosphorylation of ERK,JNK and p38MAPK,and it has been reported that LPS stim-ulated induction of phosphorylation occurs within10–30min after LPS treatment(Jung et al.,2009b).Therefore,in this study the cells were lysed to obtain proteins for Western blot after2h paeonol treatment followed by30min of LPS(1l g/ml)stimulation.The re-sults showed that paeonol suppressed the JNK and p38signaling cascades.It has been reported that JNK and p38pathways are spe-ci?c upstream signaling pathways of in?ammatory responses and this shows that paenol speci?cally inhibited the in?ammatory sig-naling(Steeg,2003).
On the basis of collective?ndings of the study it could be con-cluded that paeonol attenuated the LPS-induced gene and protein expression of iNOS,COX-2and pro-in?ammatory cytokines via blocking NF-j B,JNK and p38MAPK pathways in BV-2and RAW264.7cell lines.Thus it could be suggested that paeonol pos-sess potential in neuro-in?ammation therapy.
Con?ict of interest statement
The authors declare that they do not have any con?ict of interest.
Acknowledgments
This research was supported by a grant from Marine Bioprocess Research Center of the Marine Biotechnology Program funded by the Ministry of Land,Transport and Maritime,Republic of Korea.
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控制软件说明书 PC端软件FTM 安装及应用 系统运行环境： 操作系统中英文Windows 98/2000/ NT/XP/WIN7/ Vista， 最低配置 CPU：奔腾133Mhz 内存：128MB 显示卡：标准VGA，256色显示模式以上 硬盘：典型安装 10M 串行通讯口：标准RS232通讯接口或其兼容型号。 其它设备：鼠标器 开始系统 系统运行前，确保下列连线正常： 1：运行本软件的计算机的RS232线已正确连接至控制器。 2：相关控制器的信号线，电源线已连接正确； 系统运行步骤： 1：打开控制器电源，控制电源指示灯将亮起。 绿色，代表处于开机运行状态；橙色代表待机状态。 2. 运行本软件 找到控制软件文件夹，点击FWM.exe运行。出现程序操作界面：
根据安装软件版本不同，上图示例中的界面及其内容可能会存在某些差别，可咨询我们的相关的售后服务人员。 上图中用红色字体标出操作界面的各部分的功能说明： 1. 菜单区：一些相关的菜单功能选择执行区。 2. 操作区：每一个方格单元代表对应的控制屏幕，可以通过鼠标或键盘的点选，拖拉的方式选择相应控制单元。 3.功能区：包含常用的功能按钮。 4.用户标题区：用户可根据本身要求，更改界面上的标题显示 5.用户图片区：用户可根据本身要求，更改界面上的图片显示，比如公司或工程相关LOGO图片。 6.附加功能区：根据版本不同有不同的附加项目。 7.状态区：显示通讯口状态，操作权限状态，和当前的本机时间，日期等。 如何开始使用 1. 通讯设置 单击主菜单中“系统配置”――》“通讯配置” 选择正确的通讯端口号，系统才能正常工作。 可以设置打开程序时自动打开串口。 2.系统配置
用友T6管理软件操作手册总账日常业务处理 日常业务流程 1、进入用友企业应用平台。 T6 双击桌面上的 如设置有密码，输入密码。没有密码就直接确定。 2、填制凭证进入系统之后打开总账菜单下面的填制凭证。如下图 丄总账［演示版】国B设畫 -二疑证 i :卜0 直接双击填制凭证，然后在弹出凭证框里点增 制单日期可以根据业务情况直接修改，输入附单据数数（可以不输），凭证摘要（在后面的匝可以选择常用摘要），选择科目直接选择（不知道可以选按F2或点击后面的一）, 输入借贷方金额，凭证完后如需继续作按增加自动保存，按保存也可，再按增加 3．修改凭证 填制凭证 证 证 证 总 ￡ ■ 凭 汇 汇 流
没有审核的凭证直接在填制凭证上面直接修改，改完之后按保存。（审核、记帐了凭 证不可以修改，如需修改必须先取消记帐、取消审核）。 4．作废删除凭证只有没有审核、记帐的凭证才可以删除。在“填制凭证”第二个菜单“制单” 下面有 一个“作废恢复”，先作废，然后再到“制单”下面“整理凭证”，这样这张凭证才被彻底删除。 5．审核凭证 双击凭证里的审核凭证菜单，需用具有审核权限而且不是制单人进入审核凭证才能审核（制单单人不能审核自己做的凭证） 选择月份，确定。 再确定。 直接点击“审核”或在第二个“审核”菜单下的“成批审核” 6．取消审核 如上所述，在“成批审核”下面有一个“成批取消审核”，只有没有记帐的凭证才可 以取消审核
7．凭证记账 所有审核过的凭证才可以记帐，未审核的凭证不能记账，在“总帐——凭证——记账” 然后按照提示一步一步往下按，最后提示记帐完成。 8．取消记帐 在“总帐”—“期末”—“对帐”菜单按“ Ctrl+H ” 系统会提示“恢复记帐前状态已被激活”。然后按“总帐”——“凭证”——“恢复 记帐前状态”。最后选“月初状态”，按确定，有密码则输入密码，再确定。 10、月末结转收支 当本月所有的业务凭证全部做完，并且记账后，我们就要进行当月的期间损益结转。 点击：月末转账并选择期间损益结转。 选择要结转的月份，然后单击“全选”。点击确定后
智能窗户控制系统软件V1.0设计说明 目录 前言 (1) 第一章软件总体设计 (1) 1.1. 软件需求概括 (1) 1.2. 定义 (1) 1.3. 功能概述 (1) 1.4. 总体结构和模块接口设计 (2) 第二章控制系统的总体设计 (3) 2.1. 功能设计 (3) 第三章软件控制系统的设计与实现 (5) 3.1. RF解码过程程序设计介绍 (5) 3.2. RF对码过程设计 (6) 3.3. 通信程序设计 (8) 3.4. IIC程序设计介绍 (9) 3.5. 接近开关程序设计 (12) 3.6. 震动开关检测程序设计 (13) 3.7. 墙面按键程序设计 (15) 第四章智能窗户控制系统的设计 (17) 第五章实测与结果说明 (18) 第六章结论 (18)
前言 目的 编写详细设计说明书是软件开发过程必不可少的部分，其目的是为了使开发人员在完成概要设计说明书的基础上完成概要设计规定的各项模块的具体实现的设计工作。 第一章软件总体设计 1.1.软件需求概括 本软件采用传统的软件开发生命周期的方法，采用自顶向下，逐步细化，模块化编程的软件设计方法。 本软件主要有以下几方面的功能 (1)RF遥控解码 (2)键盘扫描 (3)通信 (4)安全检测 (5)电机驱动 1.2.定义 本项目定义为智能遥控窗户系统软件。它将实现人机互动的无缝对接，实现智能关窗，遥控开关窗户，防雨报警等功能。 1.3.功能概述 1.墙体面板按键控制窗户的开/关 2.RF遥控器控制窗户的开/关 3.具有限位，童锁等检测功能 4.实时检测大气中的温湿度，下雨关窗 5.具有防盗，防夹手等安全性能的检测
NC系统培训手册 编制单位：用友软件股份有限公司 中央大客户事业部 目录 一、NC系统登陆 .................................... 二、消息中心管理................................... 三、NC系统会计科目设置 ............................ 四、权限管理....................................... 五、打印模板设置................................... 六、打印模板分配................................... 七、财务制单....................................... 八、NC系统账簿查询 ................................ 九、辅助余额表查询................................. 十、辅助明细账查询................................. 十一、固定资产基础信息设置......................... 十二、卡片管理..................................... 十三、固定资产增加................................. 十四、固定资产变动................................. 十五、折旧计提..................................... 十六、折旧计算明细表...............................
门禁考勤管理软件 使 用 说 明 书
软件使用基本步骤
一．系统介绍―――――――――――――――――――――――――――――2二．软件的安装――――――――――――――――――――――――――――2 三．基本信息设置―――――――――――――――――――――――――――2 １）部门班组设置―――――――――――――――――――――――――3 ２）人员资料管理―――――――――――――――――――――――――3 ３）数据库维护――――――――――――――――――――――――――3 ４）用户管理―――――――――――――――――――――――――――3 四．门禁管理―――――――――――――――――――――――――――――4 １）通迅端口设置―――――――――――――――――――――――――4２）控制器管理――――――――――――――――――――――――――4３）控制器设置――――――――――――――――――――――――――6４）卡片资料管理―――――――――――――――――――――――――11 ５）卡片领用注册―――――――――――――――――――――――――12６）实时监控―――――――――――――――――――――――――――13 五．数据采集与事件查询――――――――――――――――――――――――13 六．考勤管理―――――――――――――――――――――――――――――14 1）班次信息设置――――――――――――――――――――――――――14 2）考勤参数设置――――――――――――――――――――――――――15 3）考勤排班――――――――――――――――――――――――――――15 4）节假日登记―――――――――――――――――――――――――――16 5）调休日期登记――――――――――――――――――――――――――16 6）请假/待料登记―――――――――――――――――――――――――17 7）原始数据修改――――――――――――――――――――――――――17 8）考勤数据处理分析――――――――――――――――――――――――17 9）考勤数据汇总―――――――—――――――――――――――――――18 10）考勤明细表—―――――――――――――――――――――――――18 11）考勤汇总表――――――――――――――――――――――――――18 12）日打卡查询――――――――――――――――――――――――――18 13）补卡记录查询—――――――――――――――――――――――――19
开票端操作说明 双击桌面“博思开票”图标，单击“确定”，进入开票界面： 一、开票： 日常业务——开票——选择票据类型——增加——核对票号无误后——单击“请核对票据号”——输入“缴款人或缴款单位”——选择”收费项目”、“收入标准”——单击“收费金额”。 （如需增加收费项目，可单击“增一行”） （如需加入备注栏（仅限于收款收据）），则在右侧“备注”栏内输入即可） 确认无误后，单击“打印”——“打印” 二、代收缴款书： 日常业务——代收缴款书——生成——生成缴款书——关闭——缴款——输入“专用票据号”——保存——缴款书左上角出现“已缴款”三个红字即可。 三、上报核销： 日常业务——上报核销——选择或输入核销日期的截止日期——刷新——核销。 （注意：“欠缴金额”处无论为正或为负均不可核销，解决方法见后“常见问题”）
常见问题 一、如何作废“代收缴款书” 日常业务——代收缴款书——缴款——删除“专用票据号”和“缴款日期”——保存——作废。 二、上报核销时出现欠缴金额，无法完成核销，或提示多缴。 1、首先检查有没有选择好截止日期，选择好后有没有点击“刷新”。 2、其次检查有没有做代收缴款书。注意：最后一张缴款书的日期不得晚于选择上报核 销日期。 3、若上述方法仍无效，则可能是由于以前作废过票据而未作废缴款书。解决方法： 首先作废若干张缴款书（直到不能作废为止），然后重新做一张新的缴款书。再核销。 三、打开“博思开票”时，出现“windows socket error：由于目标机器积极拒绝，无法连接。 （10061），on API’connect’” 单击“确定”，将最下面一行的连接类型“SOCKET”更换为“DCOM”，再点“连接” 即可。 四、如何设置密码 双击桌面“博思开票”，单击登录界面的右下方“改口令”输入用户编号、新密码和确认密码，单击“确认”即可。 五、更换开票人名称或增加开票人 进入开票系统——系统维护——权限管理 1、更换开票人名称：单击“用户编码”——删除“用户名”——输入新的开票人名称 ——单击“保存用户”即可。 2、增加开票人：单击“新增用户”——输入“用户编码”和“用户名”——单击“保 存用户”——单击新增的用户编码——将右边的“权限列表如下”下面的“所有”前的小方框勾上——单击右侧“保存用户权限”。 六、重装电脑系统 1、由于博思开票软件安装在D盘，所以重装电脑系统前无需做任何备份。 2、重装系统后，打开我的电脑—D盘，将“博思软件”文件夹复制到桌面上（或U盘）。 3、将安装时预留的安装光盘放入主机，打开后找到“票据核销及管理_开票端(江西欠 缴不能上报版)”（或者进入D盘----开票软件备份目录勿删文件夹里也可找到）。双 击，按提示点击“下一步”，直到“完成”。 4、双击桌面任务栏右下角“博思开票服务器”，将其关闭（或右键点击“博思开票服 务器”——“关闭服务器”）。 （这一步若找不到“博思开票服务器”，也可以用重启电脑来代替） 5、将刚才复制到桌面（或U盘）的“博思软件”再复制粘贴回D盘，若提示“此文 件夹已包含名为博思软件的文件夹”，点击下面的“全部”。 6、双击桌面“博思开票”——输入用户编码(001)——确定。 7、确认原来的票据数据没有丢失后，将桌面（或U盘）的“博思软件”文件夹删除。
控制系统使用说明 系统针对轴流风机而设计的控制系统, 系统分为上位监视及下位控制两部分 本操作为上位监控软件的使用说明: 1: 启动计算机: 按下计算机电源开关约2秒, 计算机启动指示灯点亮, 稍过大约20秒钟屏幕出现操作系统选择菜单, 通过键盘的“↑↓”键选择“windows NT 4.0”菜单，这时系统进入WINDOWS NT 4.0操作系统，进入系统的操作画面。 2：系统操作 系统共分：开机画面、停机画面、趋势画面、报警画面、主机流程画面、轴系监测画面、润滑油站画面、动力油站画面、运行工况画面、运行记录画面等十幅画面，下面就十幅画面的作用及操作进行说明 A、开机画面： 开机： 当风机开始运转前，需对各项条件进行检查，在本画面中主要对如下指标进行检查，红色为有效： 1、静叶关闭：静叶角度在14度
2、放空阀全开：放空阀指示为0% 3、润滑油压正常 4、润滑油温正常 5、动力油压正常 6、逆止阀全关 7、存储器复位：按下存储器复位按钮，即可复位，若复位不成 需查看停机画面。 8、试验开关复位：按下试验开关按钮即可，试验开关按钮在风 机启动后，将自动消失，同时试验开关也自动复位。 当以上条件达到时，按下“允许机组启动”按钮，这时机组允许启动指示变为红色，PLC机柜里的“1KA”继电器将导通。机组允许启动信号传到高压柜，等待电机启动。开始进行高压合闸操作，主电机运转，主电机运转稳定后，屏幕上主电机运行指示变红。这时静叶释放按钮变红，按下静叶释放按钮后，静叶从14度开到22度，静叶释放成功指示变红。 应继续观察风机已平稳运行后，按下自动操作按钮，启机过程结束。 B、停机画面： 停机是指极有可能对风机产生巨大危害的下列条件成立时，PLC 会让电机停止运转： 1、风机轴位移过大
用 友 T+ 软 件 系 统 操 作 手 册版本号：v1.0
目录 一、系统登录 (3) 1.1、下载T+浏览器 (3) 1.2、软件登陆 (3) 二、基础档案设置 (5) 2.1、部门、人员档案设置 (5) 2.2、往来单位设置 (6) 2.3、会计科目及结算方式设置 (6) 三、软件操作 (9) 3.1、凭证处理 (9) 3.1.1、凭证填制 (9) 3.1.2、凭证修改 (10) 3.1.3、凭证审核 (11) 3.1.4、凭证记账 (12) 3.2、月末结转 (13) 四、日常帐表查询与统计 (14) 4.1、余额表 (14) 4.2、明细账 (15) 4.3、辅助账 (16) 五、月末结账、出报表处理 (17) 5.1、总账结账 (17) 5.2、财务报表 (20)
一、系统登录 1.1、下载T+浏览器 首次登陆需要用浏览器打开软件地址，即：127.0.0.1:8000(一般服务器默认设置，具体登陆地址请参考实际配置)，第一次登陆会提示下载T+浏览器，按照提示下载安装T+浏览器，然后打开T+浏览器，输入软件登陆地址。 ，T+浏览器， 1.2、软件登陆 按键盘上的“回车键（enter）”打开软件登陆页面，如下： 选择选择“普通用户”，输入软件工程师分配的用户名和密码，选择对应的账套，以下以demo 为例，如下图：
点击登陆，进入软件，
二、基础档案设置 2.1、部门、人员档案设置 新增的部门或者人员在系统中可按照如下方法进行维护，
2.2、往来单位设置 供应商客户档案的添加方法如下： 添加往来单位分类： 2.3、会计科目及结算方式设置会计科目：
LED-ECS编辑控制系统V5.2 用 户 手 册 目录 第一章概述 (3) 1.1LED-ECS编辑控制系统介绍 (3) 1.2运行环境 (3) 第二章安装卸载 (3) 2.1安装 (3) 2.2卸载 (5) 第三章软件介绍 (5) 3.1界面介绍 (5) 3.2操作流程介绍 (13) 3.3基本概念介绍 (21) 第四章其他功能 (25) 4.1区域对齐工具栏 (25) 4.2节目对象复制、粘贴 (26) 4.3亮度调整 (26) 第五章发送 (27) 5.1发送数据 (27) 第六章常见问题解决 (28) 6.1计算机和控制卡通讯不上 (28) 6.2显示屏区域反色或亮度不够 (29)
6.3显示屏出现拖尾现象，显示屏的后面出现闪烁不稳定 (29) 6.4注意事项 (31) 6.5显示屏花屏 (31) 6.6错列现象 (32) 6.7杂点现象 (32) 第一章概述 1.1LED-ECS编辑控制系统介绍 LED-ECS编辑控制系统，是一款专门用于LED图文控制卡的配套软件。其具有功能齐全，界面直观，操作简单、方便等优点。自发布以来，受到了广大用户的一致好评。 1.2运行环境 ?操作系统 中英文Windows/2000/NT/XP ?硬件配置 CPU:奔腾600MHz以上 内存:128M 第二章安装卸载 2.1LED-ECS编辑控制系统》软件安装很简单，操作如下：双击“LED-ECS编辑控制系统”安装程序，即可弹出安装界面，如图2-1开始安装。如图所示 图2-1 单击“下一步”进入选择安装路径界面，如图2-2，如果对此不了解使用默认安装路径即可 图2-2 图2-3 单击“完成”，完成安装过程。 2.2软件卸载如图2-2 《LED-ECS编辑控制系统V5.2》提供了自动卸载功能，使您可以方便的删除《LED-ECS编辑控制系统V5.2》的所有文件、程序组件和快捷方式。用户可以在“LED-ECS编辑控制系统V5.2”组中选择“卸载LED-ECS编辑控制系统V5.2”卸载程序。也可以在“控制面板”中选择“添加/删除程序”快速卸载。卸载程序界面如图2-4，此时选择自动选项即可卸载所有文件、程序组和快捷方式。 图2-4 第三章、软件介绍
财政票据(网络版)电子化系统 开票端 操 作 说 明 福建博思软件股份有限公司
目录 1.概述 业务流程 流程说明：
1.单位到财政部门申请电子票据，由财政把单位的基本信息设置好并审核完后，财政部门给用票单位发放票据，单位进行领票确认并入库。 2.在规定时间内，单位要把开据的发票带到财政核销，然后由财政进行审核。 系统登录 登入系统界面如图： 登录日期：自动读取主服务器的日期。 所属区划：选择单位所属区划编码。【00安徽省非税收入征收管理局】 所属单位：输入单位编码。 用户编码：登录单位的用户编码【002】 用户密码：默认单位密码为【123456】 验证码：当输入错误时，会自动换一张验证码图片； 记录用户编码：勾选系统自动把用户编码保存在本地，第二次登录不需要重新输入。 填写完正确信息，点【确定】即可登入系统。 进入系统 进入系统界面如图： 当单位端票据出现变动的时候，如财政或上级直管下发票据时，才会出现此界面:
出现此界面后点击最下方的确认按钮，入库完成。 当单位端票据无变动时，直接进入界面： 2.基本编码人员管理 功能说明：对单位开票人员维护，修改开票人名称。 密码管理 修改开票人员密码，重置等操作。 收发信息 查看财政部门相关通知等。
3.日常业务 电脑开票 功能说明：是用于开票据类型为电子化的票据。 在电脑开票操作界面，点击工具栏中的【增加】按钮，系统会弹出核对票号提示框，如图： 注意：必须核对放入打印机中的票据类型、号码是否和电脑中显示的一致，如果不一致打印出来的票据为无效票据，核对完后，输入缴款人或缴款单位和收费项目等信息，全部输入完后，点【增加】按钮进行保存当前票据信息或点【打印】按钮进行保存当前票据信息并把当前的票据信息打印出来；点电脑开票操作界面工具栏中的【退出】则不保存。 在票据类型下拉单框中选择所要开票的票据类型，再点【增加】进行开票。
用友T软件系统操作手 册 Pleasure Group Office【T985AB-B866SYT-B182C-BS682T-STT18】
用 友 T+ 软 件 系 统 操 作 手 册 版本号：目录
一、系统登录 、下载T+浏览器 首次登陆需要用浏览器打开软件地址，即：(一般服务器默认设置，具体登陆地址请参考实际配置)，第一次登陆会提示下载T+浏览器，按照提示下载安装T+浏览器，然后打开T+浏览器，输入软件登陆地址。 ，T+浏览器， 、软件登陆 按键盘上的“回车键（enter）”打开软件登陆页面，如下： 选择选择“普通用户”，输入软件工程师分配的用户名和密码，选择对应的账套，以下以demo为例，如下图： 点击登陆，进入软件， 二、基础档案设置 、部门、人员档案设置 新增的部门或者人员在系统中可按照如下方法进行维护， 、往来单位设置 供应商客户档案的添加方法如下： 添加往来单位分类： 、会计科目及结算方式设置 会计科目： 系统预置170个《2013小企业会计准则》科目，如下：
结算方式，如下： 三、软件操作 、凭证处理 填制 进入总账填制凭证菜单，增加凭证，填制摘要和科目，注意有辅助核算的会计科目， 以下为点开总账的处理流程图： 如若现金流量系统指定错误，可按照以下步骤修改： 凭证在没有审核时，可以直接在当前凭证上修改，然后点击“保存”完成修改； 凭证审核 进入总审核凭证菜单下，如下图： 选择审核凭证的会计期间： 、凭证记账 进入凭证菜单下的记账菜单， 、月末结转 期间损益结转 四、日常帐表查询与统计 、余额表 用于查询统计各级科目的本期发生额、累计发生额和余额等。传统的总账，是以总账科目分页设账，而余额表则可输出某月或某几个月的所有总账科目或明细科目的期初余额、本期发生额、累计发生额、期末余额，在实行计算机记账后，我们建议用户用余额表代替总账。
创维液晶拼接控制系统 软件操作指南 【LCD-CONTROLLER12】 请在使用本产品前仔细阅读该用户指导书
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一、功能特点 二、技术参数 三、控制系统连接示意图 四、基本操作 五、故障排除 六、安全注意事项
一、功能特点创维创维--液晶液晶拼接拼接拼接控制器特点控制器特点 ★采用创维第四代V12数字阵列高速图像处理技术 视频带宽高达500MHZ，应用先进的数字高速图像处理算实时分割放大输入图像信号，在多倍分割放大处理的单屏画面上，彻底解决模/数之间转换带来的锯齿及马赛克现象，拼接画面清晰流畅，色彩鲜艳逼真。 ★具有开窗具有开窗、、漫游漫游、、叠加等功能 以屏为单元单位的前提下，真正实现图像的跨屏、开窗、画中画、缩放、叠加、漫游等个性化功能。 ★采用基于LVDS 差分传送技术差分传送技术，，增强抗干扰能力 采用并行高速总线连接技术，上位控制端发出命令后，系统能快速切换信号到命令指定的通道，实现快速响应。 采用基于LVDS 差分传送技术，提高系统抗干扰能力，外部干扰对信号的影响降到了最低，并且，抗干扰能力随频率提高而提升。★最新高速数字阵列矩阵通道切换技术 输入信号小于64路时，用户不需要再另外增加矩阵，便可以实现通道之间的任意换及显示。 ★断电前状态记忆功能 通过控制软件的提前设置，能在现场断电的情况下，重启系统后，能自动记忆设备关机前的工作模式状态。 ★全面支持全高清信号 处理器采用先进的去隔行和运动补偿算法，使得隔行信号在大屏幕拼接墙上显示更加清晰细腻，最大限度的消除了大屏幕显示的锯齿现象，图像实现了完全真正高清实时处理。纯硬件架构的视频处理模块设计，使得高清视频和高分辨率计算机信号能得到实时采样，确保了高清信号的最高视频质量，使客户看到的是高质量的完美画质。
工会经费收入专用收据(1) 福州博思软件开发有限公司 2010年6月 目录 第一部分初始安 装 ....................................................... (1) 1.1系统 安装 (1) 1.2系统登录 ............................................................ (4) 第二部分组成模块介 绍 ................................................... (4) 2.1模块组 成 (4) 2.1.1票据资料 .......................................................... (5) 2.1.2用户管 理 .......................................................... (5) 2.1.3 票据领用 (6) 2.1.4电脑开票 .......................................................... (6) 2.1.5手工开 票 .......................................................... (7) 2.1.6 手工批开票 (7) 2.1.7票据查询 .......................................................... (8) 第三部分软件操 作 ....................................................... (9) 3.2用户 管理 (10)
大屏幕控制系统软件详解说明 一软件安装 安装注意事项： 非专业人事安装：安装前请先关闭防火墙（如360安全卫士，瑞星，诺盾等），等安装完并且成功启动本软件后可重新开启防火墙； 专业人事安装：先把防火墙拦截自动处理功能改为询问后处理，第一次打开本软件时会提示一个拦截信息； 安装前请校对系统时间,安装后不能在错误的系统时间下运行/启动软件,否则会使软件注册失效，这种情况下需要重新注册； Windows 7,注意以下设置 0.1)打开控制面板 0.2) 选择系统和安全 0.3) 选择操作中心 0.4) 选择更换用户帐户控制设置 0.5)级别设置，选择成从不通知 １．软件解压后，请选择双击，进入安装界面如图１，图２ 图1
图2 ２．选择键，进入下一界面如图3 图３ 3.选中项，再按键，进入下一界面如图４
图4 4.选择键，进入下一界面如图５ 图５ ５.选中项，再选择键，进入下一界面如图６
图６ ６.选择键，进入下一界面如图７ 图８ ７.选择键，软件安装完成 二软件操作 选择WINDOWS 下开始按钮，选择程序，选择Wall Control项， 点击Wall Control软件进入大屏幕控制系统软件主界面如图9所示,整个软件分为3个区，标题区，设置区，功能区
图9 1.1标题区 大屏幕控制系统软件(只有管理员才可设置此项目) 1.2设置区 1.2.1系统 高级功能：管理员登录。 产品选型：选择拼接盒型号。 定时系统：设置定时时间。 幕墙开机：开机 幕墙关机：关机 退出：退出软件系统。 1.2.2设置 串口设置：设置使用的串口参数。 矩阵设置：设置矩阵的相关参数。 幕墙设置：幕墙设置参数。 幕墙颜色：幕墙颜色设置。 标志设置：更改幕墙名称。 系统设置：控制软件系统设置。 1.2.3工具 虚拟键盘：虚拟键盘设置。 硬件注册：可以通过时钟IC注册处理器的使用权限。 1.2.4语言 中文选择:选择软件语言类型为中文。 Ｅnglish:选择软件语言类型为英语。
用友财务管理系统操作手册 北京用友政务软件有限公司 2011年05月25日
一、账务系统： 流程：1、初始化设置及期初数装入=》2、凭证录入=》 3、凭证审核=》 4、凭证记账=》 5、月结 1、初始化设置: (1)、用自己的用户名登录【账务管理系统】=》 点击界面右边【基础资料】前的【+】号=》点击【会计科 目】前的【+】号=》双击【建立会计科目】=》设置会计科 目及挂接辅助账。(2)、点击界面右边【账务】前的【+】号 =》点击【初始建账数据】前的【+】号=》双击【期初余额 装入】=》点击【确定】=》然后对期初数据进行录入 2、凭证录入:用自己的用户名登录【账务管理系统】=》点击界 面右边【账务】前的【+】号=》点击【凭证管理】前的【+】 号=》双击【编制凭证】=》然后在【编制凭证】界面录入 收入/支出的凭证。 3、凭证审核：点击界面右边【账务】前的【+】号=》点击【凭 证管理】前的【+】号=》双击【凭证处理】=》选中需要审 核凭证的日期=》在左下角选择凭证的状态【未审核】=》 点击右键全选=》点击【审核】； 4、凭证记账：点击界面右边【账务】前的【+】号=》点击【期 末处理】前的【+】号=》双击【凭证处理】=》选中需要记 账凭证的日期=》在左下角选择凭证的状态【已审核】=》 点击右键全选=》点击【记账】； 5、月结：点击界面右边【账务】前的【+】号=》点击【期末
处理】前的【+】号=》双击【期末处理向导】=》点击【结 账向导】=》全部点击【下一步】=》下到最后点击【完成】 二、电子系统： 1、输出单位资产负债表：双击【电子报表系统】=》【管理员】 登录=》在右上角【报表数】下点击【基本户】/【专账一】 /【专账二】下前的【+】号=》双击【资产负债表】=》点击 最右上面【数据】下=》=》点击【登录数据库】=》双击【账 务系统】=》用自己的用户进行登录=》如果图片闪烁就证 明已经登录=》点击【退出】=》点击最右上角找到【插入】 功能菜单=》点击【表页】=》选择出报表的最后日期(如1 月：则时间2011年1月31日)=》选择复制指定表页 =》点击放大镜=》选择【本公司】=》选中【格式】点击【确定】=》在点【确定】=》左 下角有【第201101期】=》点击编制【眼睛图标】。=》调 试报表=》点击【保存】=》打印报表。 2、输出单位支出明细表：双击【电子报表系统】=》【管理员】 登录=》在右上角【报表数】下点击【基本户】/【专账一】 /【专账二】下前的【+】号=》双击【支出明细表】=》点击 最右上面【数据】下=》=》点击【登录数据库】=》双击【账 务系统】=》用自己的用户进行登录=》如果图片闪烁就证 明已经登录=》点击【退出】=》点击最右上角找到【插入】
目录 1，系统概述--------------------------------------------------------------------------------------------------1 1.1 系统简介---------------------------------------------------------------------------------------------2 1.2 系统主要组成---------------------------------------------------------------------------------------2 1.3 系统硬件简要连接图------------------------------------------------------------------------------3 1.4 实际连线图------------------------------------------------------------------------------------------3 2,系统软件使用软件简要说明-----------------------------------------------------------------------------5 2.1 介绍---------------------------------------------------------------------------------------------------5 2.2 操作步骤---------------------------------------------------------------------------------------------5 2.3 取景窗口---------------------------------------------------------------------------------------------7 2.4 flash/cel文件的播放--------------------------------------------------------------------------------7 注1:连接网络的相关设置修改--------------------------------------------------------------9 注2:本机IP的查询----------------------------------------------------------------------------9 注3:本机IP的修改----------------------------------------------------------------------------10 注4:控制器IP的修改-------------------------------------------------------------------------11 3,对应表制作与选择-----------------------------------------------------------------------------------------12 3.1 介绍---------------------------------------------------------------------------------------------------12 3.2 操作步骤---------------------------------------------------------------------------------------------12 4,说明-----------------------------------------------------------------------------------------------------------14 4.1 ONC1A------------------------------------------------------------------------------------------------14 4.2 ONC1B------------------------------------------------------------------------------------------------14 4.3 ONC1C------------------------------------------------------------------------------------------------15 4.4 ONC1D------------------------------------------------------------------------------------------------15 4.5 ONC1E------------------------------------------------------------------------------------------------16 4.6 ONC1F------------------------------------------------------------------------------------------------17 4.7 ONC1G------------------------------------------------------------------------------------------------17 4.8 ONC1F------------------------------------------------------------------------------------------------17 5,附件-----------------------------------------------------------------------------------------------------------19 5.1 数码按钮控制板说明--------------------------------------------------------------------------------19 5.2 象素点排列说明--------------------------------------------------------------------------------------19
政府采购网上公开信息系统供应商操作指南 福建博思软件股份有限公司 2017年06月
目录 1. CA办理 (4) 1.1 CA办理 (4) 1.2 CA盖章 (4) 2. 系统注册 (4) 2.1系统注册 (4) 2.2 注册成功，供应商登录 (5) 3.系统基础操作 (6) 3.1 CA控件下载 (6) 3.2进行政府采购活动 (9) 3.3 供应商资料维护 (10) 3.4 供应商项目报名 (11) 3.5供应商用户管理 (11) 4．投标流程 (11) 4.1项目报名 (11)
4.2 投标文件编制 (13) 4.2.1客户端安装 (13) 4.2.2投标客户端路径修改 (15) 4.2.3应答 (16) 4.2.4标书加密 (21) 4.2.5接着点击退出系统 (21) 4.3投标文件上传： (22) 4.4合同签订： (23) 4.5验收申请： (23)
1.CA办理 供应商须办理福建省CA，进行政府采购活动 1.1CA办理 可登陆https://www.doczj.com/doc/bb279323.html,/或者联系客服0591-968975。 1.2CA盖章 CA盖章操作系统为：XP(SP2)不支持（浏览器目前测试都支持）。 2. 系统注册 2.1系统注册 登入福建省政府采购网https://www.doczj.com/doc/bb279323.html,/，找到登陆与注册进行供应商注册
2.2 注册成功，供应商登录 注：原省网已注册的供应商已完成迁移，请各供应商使用注册时的组织机构代码登录，密码初始为1。
3.系统基础操作 3.1 CA控件下载与安装 登录之前可先进行CA控件下载 (1)打开ISignature,点击installer.exe进行安装
用友财务软件操作流程手册 系统管理 一增加操作员 1、系统管理→系统→注册→输入用户名（admin）→无密码→确定 2、单击权限→操作员→点增加→输入编号、姓名、口令→点增加 二、建新账套 1、系统管理→系统→注册→输入用户名（admin）→无密码→确定 2 单击帐套→建立→输入帐套号、帐套名称、→设置会计期间→下一步→ 输入单位名称→下一步→选择企业类型（工业类型比商业类型多产成品入库单，和材料出库单）→行业性质→选择帐套主管→在“行业性质预置科目”前面打钩则系统将预置所选行业会计科目（否则不予预置）→下一步→如需分类在项目前面方框内打钩→下一步→完成 三、分配权限 1、系统管理→系统→注册→输入用户名（admin）→无密码→确定 2、赋权限的操作顺序： A 受限，明细权限设置权限”-→“权限”菜单→首先选择所需的账套→选操作员→点增加 B 帐套主管权限设置选择所需帐套→再选操作员→在帐套主管前面直接打钩 四、修改账套 1、以“账套主管（不是admin）”身份进入“系统管理”模块（进入系统服务→系统管理→注册） 2、单击帐套→修改 五．备份 打开系统管理→系统→注册admin →帐套→备份→选择存放路径 六．恢复 系统管理→系统→注册→admin →帐套→恢复（选择本分文件的路径，lst为后缀名的文件）总帐系统 初始化 一、启用及参数设置 二、设置“系统初始化”下的各项内容（其中：最后设置会计科目和录入期初余额，其余各项从上向 下依次设置） 1 会计科目设置 1、指定科目 系统初始化→会计科目→编辑（菜单栏中的）→指定科目 现金总帐科目把现金选进以选科目 银行总帐科目把银行存款选进已选科目
动环监控软件操作 手册
深圳市通讯威科技有限公司 EP-MEVP SYSTEM 动力环境集中监控系统 安装使用说明书 版本 2.0
目录 第一章软件的安装卸载升级 ................................. 错误!未定义书签。 1.1软件安装对计算机的配置要求........................ 错误!未定义书签。 1.2软件的安装 ....................................................... 错误!未定义书签。 1.3软件的卸载 ....................................................... 错误!未定义书签。 1.4软件的升级 ....................................................... 错误!未定义书签。第二章软件的基本操作 .......................................... 错误!未定义书签。 2.1登录和进入软件操作界面................................ 错误!未定义书签。 2.2添加/设置/修改/删除硬件设备以及参数设置错误!未定义书签。 2.2.1添加/设置控制器、采集器参数................. 错误!未定义书签。 2.2.2修改/删除硬件设备 .................................... 错误!未定义书签。 2.3监控设置及记录查询 ....................................... 错误!未定义书签。 2.3.1报警方式定义 ............................................. 错误!未定义书签。 2.3.2语音电话报警 ............................................. 错误!未定义书签。 2.3.3短信报警 ..................................................... 错误!未定义书签。 2.3.4监控实时记录 ............................................. 错误!未定义书签。 2.3.5监控报警记录 ............................................. 错误!未定义书签。 2.3.6温湿度数据记录.......................................... 错误!未定义书签。 2.3.7 UPS监控数据记录 ...................................... 错误!未定义书签。 2.3.8 电话短信报警数据记录 .............................. 错误!未定义书签。 2.3.9 空调监控数据记录...................................... 错误!未定义书签。