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歲歲又重陽范文第1篇

【摘要】

目的：檢測視網(wǎng)膜色素上皮細(xì)胞對骨髓間充質(zhì)干細(xì)胞分化的調(diào)節(jié)作用。方法：體外培養(yǎng)人骨髓間充質(zhì)干細(xì)胞（BMSC）和視網(wǎng)膜色素上皮（RPE）細(xì)胞。RPE細(xì)胞采用紫外線照射處理，而后與CFSE標(biāo)記的BMSC共培養(yǎng)14d。并在共培養(yǎng)體系中加入牛眼視網(wǎng)膜提取物（BRE）以研究視網(wǎng)膜成分對此分化過程的影響。采用NSE， Nestin和GFAP抗體標(biāo)記檢測BMSC分化前后的表達(dá)特征。結(jié)果：BMSC在與RPE細(xì)胞共培養(yǎng)后，能夠分化成為神經(jīng)樣細(xì)胞，并表達(dá)神經(jīng)性細(xì)胞的特異性標(biāo)記NSE，Nestin和GFAP。BRE能夠顯著促進(jìn)共培養(yǎng)體系中BMSC向神經(jīng)樣細(xì)胞的分化。結(jié)論：RPE和BRE能夠誘導(dǎo)BMSC分化為神經(jīng)樣細(xì)胞。

【關(guān)鍵詞】 髓間充質(zhì)干細(xì)胞；視網(wǎng)膜色素上皮細(xì)胞；神經(jīng)樣細(xì)胞；牛眼視網(wǎng)膜提取物

AbstractAIM: To detect the differentiation effects of retinal cells or extracts on bone marrowderived mesenchymal stem cells (BMSC).METHODS: Human fetal BMSC were previously labeled by carboxyfluorescein succinimidyl ester (CFSE)， and cocultured with retinal pigment epithelial （RPE） cells which were pretreated with ultraviolet irradiation at a ratio of 1∶1 to induce the differentiation of BMSC for up to 14 days. In some assays， a retinal extract of bovine retinal extract (BRE) was added to detect the potential effects of retinal component on the differentiation of BMSC. In addition， neuronspecific enolase (NSE)， Nestin and Glial fibrillary acidic protein (GFAP) immunostaining were performed to determine the characteristics of BMSC.RESULTS: The results indicated that by cocultured with RPE cells， fetal BMSC were differentiated into neurallike cells expressing special neuronal markers Nestin， GFAP and NSE. And the expression of these markers was obviously increased by BRE. CONCLUSION: Retina derived cells and extracts can induce the differentiation of BMSC into neurallike cells.

KEYWORDS: bone marrowderived mesenchymal stem cells; retinal pigment epithelial cells; neurallike cells; bovine retinal extract

INTRODUCTION

According to the origin and potential of differentiation， two types of stem cells can be distinguished: embryonic stem cells (ESC) and somatic stem cells (SSC). SSC are isolated from fetal (after gastrulation) or adult tissues， but classically the differentiation fate of these cells are believed to be limited to the cell types that belong to the tissue from which they originate. However， previous studies have suggested that these tissuespecific stem cells might be able to differentiate into cell types of other lineages[1]. Bone marrowderived mesenchymal stem cells (BMSC) are one of the major subpopulations of SSC， which is extensively studied with respect to transdifferentiation potential[2]. Recent studies have described that BMSC can be differentiated into neurallike cells in vitro under specific induced culture conditions[3，4] or developed spontaneously differentiation under noninduced standard culture conditions[57]. Usually the differentiation is induced just by addition of growth factors and/or chemicals in culture medium. The agents include βmercaptoethanol and dimethylsulfoxide， epidermal growth factor (EGF) and brainderived neurotrophic factor (BDNF)， isobutylmethylxanthine/dibutyryl cAMP， or 5AzaC/growth factors. Moreover， a small proportion of BMSCderived cells differentiated into neuronlike cells expressing NeuN and glial cells expressing Glial fibrillary acidic protein (GFAP) when cocultured with rat fetal mesencephalic or striatal cells. The retinal pigment epithelium(RPE) is a hexagonally packed monolayer cell in ocular retina that performs critical functions in the maintenance of the physiology of photoreceptors which is developed from the outer layer of the optic cup. The research of Chiou et al[8] shows that coculture of human BMSC with human retinal pigment epithelium (HRPE) cells facilitates the retina and photoreceptorlineage differentiation of adult human BMSC. This system indicates the retinal differentiation potential of BMSC. In this study， we established a coculture fetal BMSC/RPE system by directly mixing them and showed that under such condition， fetal BMSC was differentiated into neural phenotypelike cells and expressed neuronal markers such as Nestin， GFAP and neural specific enolase[5]. Besides， our results demonstrated that bovine retinal extract (BRE) could markedly promoted the neurallineage differentiation of fetal BMSC in vitro.

MATERIALS AND METHODS

Materials Human fetal BMSC were cultured in this assay. The use of human tissues in this study was approved by the Shandong Eye Institute Medical Ethics Committee and was in compliance with the Declaration of Helsinki. In order to eliminate unwanted types of cells presented in the marrow aspiration， mononuclear cells were obtained by Ficoll Hypaque density gradient centrifugation (lymphoprep， 1073g/L; TBD， China) and BMSC were selected by plastic adhesion. Briefly， a small percentage of cells isolated from the density interface of 1073g/L were seeded in 6 well plates using the medium of LG DMEM supplemented with 100mL/L FBS (Gibco， USA). After 3 days undisturbed to promote cell attachment， the nonadherent cells were removed by changing the medium. At nearconfluence， about two weeks after initial plating， cells were subcultured after trypsin digestion. Fetal BMSC were isolated by Ficoll Hypaque density gradient centrifugation and the physical property of adherence to plastic culture dishes were confirmed. BMSC cultured in our experiment grew as fibroblastic cells with scant cytoplasm and contain granules around nuclei， which were similar to the cells described in previous reports[8]. Fetal RPE cells were isolated and cultured as described previously[9] and these cells showed typical polygonal shape. Briefly， sheets of RPE were dissected from the choroids of fetal eye cups and cultured in lowcalcium (0.05mmol/L Ca2+) MEM (Sigma， St. Louis， MO; catalog number M8028) medium containing proposed supplements according to the reports of Hu and Bok [9]. Floating RPE cells were collected and passaged with normalcalcium medium (Sigma， St. Louis， MO; catalog number M2279) containing the same supplements.

BMSC/RPE 　coculture Firstly， fetal BMSC at 35 passage were labeled with CFSE (PKCA705C375， Probior GmbH) in order to be distinguished from RPE cells. A total of 106 to 107 cells were washed twice with phosphate buffered saline (PBS) and then incubated with 2.5μmol/L CFSE in PBS for 10 minutes in dark at room temperature. Then cells were washed twice with media containing 100mL/L FBS. Secondly， the second passage of fetal RPE cells were pretreated under a UV lamp for 15 minutes to abolish its selfroliferation as the method of Chiou et al[8] and washed three times with DHanks. Then the inactivated cells were digested with 2.5g/L trypsin0.2g/L EDTA and resuspended in medium containing 100mL/L FBS. Lastly， 2×105 of fetal BMSC and RPE cells prepared as above were mixed together and plated on gelatincoated 24wells plates for up to 14 days. In some assays， 10μg BRE was added in the medium. BRE was prepared by homogenizing 12 fresh bovine retinas per 100mL CMFBSS and stirring overnight in the dark at 4℃. The mixture was cleared by centrifugation (12000g for 20 minutes) and the supernatants were collected[9]. The noninduced fetal BMSC were cultured as the control group. In each group four duplicate wells were set up， and the assay was repeated for 3 times independently.

Immunocytochemistry Cells were fixed at 20℃ with cold methanol for 5 minutes. For staining， samples were rehydrated in PBS/2mL/L Triton X100. Nonspecific binding was blocked with 100mL/L normal serum in PBS/2mL/L Triton X100 for 30 minutes at room temperature. Cells were then incubated overnight at 4℃ with the following primary antibodies diluted in blocking solution， polyclonal rabbit antinestin (1∶100， Boster， China); polyclonal rabbit antiNSE (1∶100， Boster， China); polyclonal rabbit antiGFAP (ready to use， Beijing Zhongshan Golden Bridge Biotechnology CO， LTD， China). Rhodamine conjugated Goat anti rabbit IgG (1∶100; Beijing Zhongshan Golden Bridge Biotechnology CO， LTD， China) was used as secondary antibody. After washing with PBS for 3 times， samples were counterstained with DAPI (Santa Cruz， USA). Negative controls were performed by omitting primary antibodies.

RESULTS

Differentiation of BMSC In this assay， we detected the differentiation characteristics of BMSC cocultured with inactivated RPE cells. In order to distinguish BMSC from RPE cells， BMSC were labeled with CFSE before coculture. After 3 days， we observed that some of cytoplasm of cultured cells retracted toward the nucleus， presenting a more spherical shape and extending processes; the changed cells were CFSEpositive demonstrating the BMSC origin(Figure 1). Furthermore， the cocultured BMSC aggregated into neurospherelike body and cells with a spindleshaped morphology were observed 10 days later (Figure 2). Immunocytochemistry was performed to investigate the expression of neuronal markers， including Nestin (neural precursor marker)， NSE (neuronal marker) ， and GFAP (astroglial maker). These analysis revealed that BMSC expressed the neural makers when cocultured with RPE. On the contrary， no neural marker was found in untreated fetal BMSC (Figure 3).Figure 1The morphological changes of fetal BMSC after induced for 3 days A and D， BMSC cocultured with RPE cells without addition of BRE; B and E， BMSC cocultured with RPE cells with addition of BRE; C， untreated BMSC. A， B and C， phasecontrast photomicrographs; D and E， confocal photomicrographs presenting the CFSEstaining cells， ×100 （略）Figure 2The morphological changes of fetal BMSC after induced for 10 days A and D， BMSC cocultured with RPE cells without addition of BRE; B and E， BMSC cocultured with RPE cells addition of BRE; C， untreated BMSC. A， B and C， phasecontrast photomicrographs; D and E， confocal photomicrographs presenting the CFSEstaining cells， ×100 （略）Figure 3Nestin， NSE and GFAP expression of BMSC upregulated in BRE treated group after induced for 14 days(Bar=100μm)A， D and G， +RPE cells， BRE; B， E and H， +RPE cells +BRE; C， F and I， untreated（略）

Effects of BRE In some assay， 10μg BRE was added to the induction system. The neuronlike morphology of BMSC treated with BRE in the medium was induced much more markedly at the early induction stage (Figure 1). CFSE is used to fluorescently label live cells and is equally partitioned to daughter cells during pision and can be used to measure cell proliferation. After 10 days， the CFSE intensity of the cells was markedly decreased by addition of BRE in culture medium(Figure 2)， suggesting that BRE may promote the proliferation of the cells. The immunoreactivity for specific neural markers was increased obviously (Figure 3). Especially， the expression of NSE and GFAP was significantly upregulated as compared with the group without BRE.

DISCUSSION

In this study， we detected the effects of RPE cells and BRE on the differentiation of BMSC to determine the potential induction function of retinaderived components on BMSC. Before induction， fetal BMSC had a flat， elongated， spindlelike structure， similar to that of fibroblasts. After induction， the CFSE positive BMSC showed the morphological characteristics of neuronal cells such as long multipolar extensions and branching ends after 3 days; and then aggregated into spheroid 10 days later. And the neural lineage differentiation of BMSC was demonstrated by the expression of some specific neural markers detected by immunocytochemistry assay. In addition， BRE could promote the neurallike cells differentiation of BMSC in this coculture system. We didnt get the similar results of differentiation BMSC into retinal lineage cells as Chiou et al by coculture of BMSC with RPE cells in this experiment. This may be attributed to the existence of some differences between our induction methods. Chiou et al[8] first induced the BMSC to a spheroid body using neurogenic selection medium for 2 weeks， and then followed a further differentiation in the medium containing RPE cells as a feeder layer for another 23 weeks. However， we directly combined the undifferentiated BMSC and RPE cells together and induced differentiation for 2 weeks. Considering the neural ectoderm developing origin of RPE， RPE cells may play a role in inducing BMSC into neurallineage cells.

Previous reports seemed that the protein of BRE has the function of maintenance the proliferation and differentiation of RPE cells[9]. In order to detect whether BRE could have some function in BMSC differentiation， 10μg protein/mL of BRE was used. The results revealed that BRE could promote a higher expression of nestin， an intermediate filament protein that is predominantly expressed during neural development to some degree[10]. Especially， BRE also enhanced the expression of NSE， a unique form of the glycolytic enzyme enolase found in neurons and in virtually all of the neuroendocrine， paraneuronal cell types， and GFAP， a glial protein that is found in glial cells such as astrocytes. These results indicated that BRE could promote the neural differentiation of BMSC. Given that the retina is developed from neural ectoderm and considering that BRE contains prominent portion of intracellular proteins which do not release and contact cell during retinal development; our results suggested that some of the retinal component must have functions in inducing the differentiation of neural lineage cells. Interesting， these neurallike cells differentiated from BMSC in this assay are both NSE and GFAP positive， we suppose that these neurallike cells are neuron/astrocyte precursor cells which may further differentiated into neuron or astrocyte cells. In conclusion， BMSC from fetal bone marrow were differentiated into neurallike cells expressing the special markers of neural cells by coculture with RPE cells， and the component from retinal may promote BMSC changing into neural lineage cells.

參考文獻(xiàn)

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歲歲又重陽范文第2篇

【摘要】 目的 觀察血管緊張素Ⅱ聯(lián)合5-氮雜胞苷體外誘導(dǎo)骨髓間充質(zhì)干細(xì)胞 (BMMSCs) 定向分化為心肌樣細(xì)胞的作用。方法 采用密度梯度離心法分離大鼠BMMSCs，取第3代BMMSCs進(jìn)行分組誘導(dǎo):血管緊張素Ⅱ組(AngⅡ，終濃度為0.1μmol/L)、5-氮胞苷組 (5-aza，終濃度為10μmol/L)、AngⅡ聯(lián)合 5-aza組 (終濃度分別為0.1μmol/L與10μmol/L)、空白對照組。誘導(dǎo)24h后更換常規(guī)培養(yǎng)液繼續(xù)培養(yǎng) 4 周。倒置相差顯微鏡觀察細(xì)胞的形態(tài)學(xué)變化，MTT法檢測細(xì)胞活性及增殖能力，免疫熒光染色法鑒定誘導(dǎo)后BMMSCs中心肌特異性肌鈣蛋白 I (cTnI) 的表達(dá)，流式細(xì)胞計(jì)數(shù)法計(jì)算心肌樣細(xì)胞誘導(dǎo)率， 透射電鏡觀察誘導(dǎo)后心肌樣細(xì)胞的超微結(jié)構(gòu)。結(jié)果 原代培養(yǎng)的BMMSCs14天形成集落， 傳代細(xì)胞體積變大， 誘導(dǎo)后細(xì)胞呈長梭形，呈一致性生長， 并出現(xiàn)肌島樣結(jié)構(gòu)。MTT法顯示AngⅡ聯(lián)合 5-aza組細(xì)胞增殖能力優(yōu)于AngⅡ組與5-氮胞苷組。免疫熒光染色結(jié)果顯示誘導(dǎo)后BMMSCs 表達(dá)心肌特異性蛋白cTnI。流式細(xì)胞計(jì)數(shù)法顯示：AngⅡ組、5-aza 、AngⅡ聯(lián)合 5-aza組心肌樣細(xì)胞誘導(dǎo)率分別為(25.3±2.2)%、(24.6±1.9)%、(30.0±1.7)%，AngⅡ聯(lián)合 5-aza組心肌樣細(xì)胞誘導(dǎo)率高于AngⅡ組與5-氮胞苷組(P>0.05)。透射電鏡可見平行排列的肌絲、Z線樣物質(zhì)。結(jié)論 血管緊張素Ⅱ聯(lián)合5-氮雜胞苷可在體外誘導(dǎo)大鼠BMMSCs定向分化為心肌樣細(xì)胞，其誘導(dǎo)分化率高于單純5-氮雜胞苷誘導(dǎo)。

【關(guān)鍵詞】 血管緊張素Ⅱ; 5-氮雜胞苷; 骨髓間充質(zhì)干細(xì)胞;心肌樣細(xì)胞

[Abstract] Objective To observe the effect of the differentiation of bone marrow mesenchymal stem cells(BMMSCs) into cardiomyocyte-like cells with 5-azacytidine and angiotensinⅡ.Methods BMMSCs were isolated from bone marrow of SD mouse by density gradient centrifugation. The third passage cells were pided into four groups:AngⅡgroup (0.1μmol/ L)，5-aza group(10μmol/L)，AngⅡ combined with 5-aza group (0.1μmol/L and 10μmol/L)，untreated group as control. After 24h induction， the medium was changed to complete culture medium without any inductor and the cells were culture for 4 weeks. The morphological changes were observed under phase contrast microscope. The effect of AngⅡ and 5-aza on the BMMSCs proliferation was observed with methyl thiazolyl tetrazolium (MTT) assay. The cardiomyogenic cells were identified by immunofluorescence staining. The differentiation ratio was examined by flow cytometer. The ultrastructures of the induced cells were viewed with a transmission electron microscope.Results BMMSCs of primary culture formed cell colonies at 14 days. The passaged cells were larger than those of primary culture. After induction， the cells presented long spindle， aligned in parallel and formed “muscle island”- like structure. MTT assay showed that the cell proliferation in AngⅡ and 5-aza group outweighed that of in AngⅡgroup or 5-aza group. The expression of specific protein of cardiac troponin I (cTnI) in induced BMMSCs was positive. Flow cytometer showed that the differentiation ratio in AngⅡgroup，5-aza group and AngⅡ combined with 5-aza group were respectively(25.3±2.2)%，(24.6±1.9)%，(30.0 ±1.7)%， demonstrating that the differentiation ratio in AngⅡ combined with 5-aza group was higher than that of in AngⅡ group or 5-aza group(P>0.05). Transmission electron microscopy showed that the induced cells had myofilaments， Z line-like substances.Conclusion AngiotensinⅡ combined with 5-Azacytidine can induce the differentiation of BMMSCs into cardiomyocyte-like cells， and the differentiation ratio was higher than that of in 5-aza only.

[Key words] angiotensin Ⅱ;5-azacytidine;bone marrow mesenchymal stem cells;cardiomyocyte-like cells

缺血性心臟病在我國的發(fā)病率逐年升高，嚴(yán)重威脅著人類的生存及生活質(zhì)量。近幾年來，細(xì)胞移植技術(shù)發(fā)展迅速，借助此技術(shù)可以在壞死部位移植具有收縮功能的細(xì)胞，為治療缺血性心臟病提供了新的方法，也成為治療缺血性心臟病的研究熱點(diǎn)課題。隨著研究的深入，選用的靶細(xì)胞目前多集中于有多向分化能力的各種干細(xì)胞，其中研究的較多的是骨髓間充質(zhì)干細(xì)胞(bone marrow mesenchymal stem cells， BMMSCs)[1]，有研究顯示BMMSCs可以被化學(xué)物質(zhì) 5-氮雜胞苷 (5-azacytidine， 5-aza)誘導(dǎo)為心肌樣細(xì)胞[2，3]。也有研究顯示，血管緊張素Ⅱ在體外可能經(jīng)細(xì)胞外信號調(diào)節(jié)激酶通路誘導(dǎo)人BMMSCs向心肌樣細(xì)胞分化[4]，提示血管緊張素Ⅱ?qū)MMSCs的分化發(fā)揮重要作用。于是我們設(shè)想：血管緊張素Ⅱ聯(lián)合5-氮雜胞苷共同誘導(dǎo)能否提高BMMSCs向心肌細(xì)胞的分化率 ?與單純血管緊張素Ⅱ及單純 5-aza的誘導(dǎo)作用相比又有何差別?本實(shí)驗(yàn)即通過聯(lián)合誘導(dǎo)培養(yǎng)，觀察血管緊張素Ⅱ聯(lián)合5-氮雜胞苷對 BMMSCs向心肌細(xì)胞分化所起的作用，并與傳統(tǒng)誘導(dǎo)分化劑 5-aza比較。

1 材料與方法

1.1 主要試劑與實(shí)驗(yàn)動物 AngⅡ ( Sigma， USA)， 5-azacytidine ( Sigma， USA)，L-DMEM 培養(yǎng)基(Hyclone， USA)，胎牛血清(杭州四季青)， 胰蛋白酶(Sigma， USA)， Ficoll 淋巴細(xì)胞分離液(1.077g/ml，TBD公司)， MTT (Sigma，USA)，DMSO (Sigma，USA)，羊抗鼠cTnI抗體(Santa cruz)，F(xiàn)ITC標(biāo)記的抗羊IgG (Boster)，Hoechst33258 (Sigma， USA)。健康SD 大鼠，4周齡，質(zhì)量80～100g，雄性，由第四軍醫(yī)大學(xué)動物實(shí)驗(yàn)中心提供。

1.2 方法

1.2.1 BMMSCs的體外分離和培養(yǎng) 頸椎脫臼法處死大鼠，取其四肢長骨，75%酒精浸泡5min。無菌條件下分離出脛骨和股骨，剪除兩端骨骺，用不含血清的DMEM培養(yǎng)液沖洗骨髓腔，充分混勻。將細(xì)胞懸液緩慢加入含淋巴細(xì)胞分離液的離心管中，2000r/min， 離心20min。取中間云霧狀有核細(xì)胞層， 加入L-DMEM不完全培養(yǎng)液，充分混勻后，1500r/min， 離心10min，棄上清，加入完全培養(yǎng)液，充分混勻后，調(diào)整細(xì)胞密度為1×106/ml，接種于T-25塑料培養(yǎng)瓶， 37℃含體積分?jǐn)?shù)為0.05的CO2孵箱中飽和濕度培養(yǎng)。3天后首次換液， 除去不貼壁的懸浮細(xì)胞，以后每3天換液1次。待細(xì)胞長到80%融合時(shí)， 用2.5g/L胰蛋白酶進(jìn)行消化，按1：2的比例進(jìn)行傳代。體外培養(yǎng)至第3代備用。

1.2.2 BMMSCs的誘導(dǎo)分化 將傳至第3代的BMMSCs進(jìn)行分組誘導(dǎo)，共分為4組：(1)AngⅡ組(終濃度為0.1μmol/L);(2)5-aza組(終濃度為10μmol/L);(3)AngⅡ加5-aza組(終濃度分別為0.1μmol/L、10μmol/L);(4)空白對照組， 僅用基本培養(yǎng)基誘導(dǎo)。各組誘導(dǎo)24h后更換完全培養(yǎng)基繼續(xù)培養(yǎng)，每3天換液1次， 并觀察細(xì)胞形態(tài)變化， 連續(xù)培養(yǎng)4周。

1.2.3 形態(tài)學(xué)鑒定 每日在倒置顯微鏡下觀察誘導(dǎo)前、后細(xì)胞生長和形態(tài)變化。

1.2.4 MTT法檢測細(xì)胞活性并描繪細(xì)胞增殖曲線 將第3代BMMSC以1×105/ml接種于4個(gè)96孔板中， 每塊板選取24孔，分為4組，每組6孔，每孔加200μl細(xì)胞懸液。培養(yǎng)24h后吸棄孔內(nèi)培養(yǎng)液，PBS洗滌2次，前3組分別加入含AngⅡ、5-aza、AngⅡ和5-aza的誘導(dǎo)培養(yǎng)液，第4組加入基本培養(yǎng)基作為對照組。誘導(dǎo)24h后各組更換完全培養(yǎng)基繼續(xù)培養(yǎng)。于培養(yǎng) 1、3、5、7天后進(jìn)行MTT 檢測，每孔加入20μl MTT(5mg/ml)，孵育4h后終止培養(yǎng)。棄孔內(nèi)上清液，每孔再加入150μlDMSO， 打勻振蕩10min，使結(jié)晶物充分溶解。選擇490nm波長，在酶聯(lián)免疫檢測儀上測定各孔光吸收值(A)，并描繪細(xì)胞增殖曲線。

1.2.5 免疫熒光細(xì)胞化學(xué)檢測 將誘導(dǎo)培養(yǎng)1、2、3、4 周的細(xì)胞進(jìn)行爬片，4%的多聚甲醛固定，3%過氧化氫-甲醇室溫孵育10min ，正常山羊血清封閉，加cTnI抗體，4℃過夜，PBS沖洗，加FITC標(biāo)記的二抗，37℃孵育40min，PBS 沖洗，Hoechst33258室溫孵育10min，PBS 沖洗，甘油封片，熒光顯微鏡觀察并拍照。

1.2.6 流式細(xì)胞計(jì)數(shù)法 2.5g/L的胰蛋白酶消化收集誘導(dǎo)4周的細(xì)胞， PBS調(diào)整細(xì)胞懸液密度約1×106個(gè)細(xì)胞 /ml， 滴加羊抗鼠 cTnI抗體，室溫 1h，再滴加 FITC標(biāo)記的抗羊 IgG，室溫避光 40min，40g/L多聚甲醛固定 20min。最后流式細(xì)胞儀測定 FITC陽性標(biāo)記細(xì)胞占細(xì)胞總數(shù)百分比。

歲歲又重陽范文第3篇

重陽節(jié)，又稱重九節(jié)、曬秋節(jié)、踏秋，中國傳統(tǒng)節(jié)日。慶祝重陽節(jié)一般會包括出游賞秋、登高遠(yuǎn)眺、觀賞、遍插茱萸、吃重陽糕、飲酒等活動。每年的農(nóng)歷九月初九日，與除夕、清明節(jié)、中元節(jié)三節(jié)統(tǒng)稱中國傳統(tǒng)四大祭祖的節(jié)日。重陽節(jié)，早在戰(zhàn)國時(shí)期就已經(jīng)形成，到了唐代被正式定為民間的節(jié)日，此后歷朝歷代沿襲至今。重陽與三月初三日踏春皆是家族傾室而出，重陽這天所有親人都要一起登高避災(zāi)。

《易經(jīng)》中把六定為陰數(shù)，把九定為陽數(shù)，九月九日，日月并陽，兩九相重，故曰重陽，也叫重九。重陽節(jié)早在戰(zhàn)國時(shí)期就已經(jīng)形成，自魏晉重陽氣氛日漸濃郁，倍受歷代文人墨客吟詠，到了唐代被正式定為民間的節(jié)日，此后歷朝歷代沿襲至今。

1989年農(nóng)歷九月九日被定為老人節(jié)，倡導(dǎo)全社會樹立尊老、敬老、愛老、助老的風(fēng)氣。2006年5月20日，重陽節(jié)被國務(wù)院列入首批國家級非物質(zhì)文化遺產(chǎn)名錄。 2016重陽節(jié)經(jīng)典賀詞

1.送你九九重陽節(jié)開心秘方：工作放一放，心事擱一擱，煩惱拋一拋，身體歇一歇，心情釀一釀，開心笑一笑，牙齒露一露，快樂好心情。祝天天開心!

2.歲歲重陽，今又重陽，日日思念，來把情傳，重陽九月九，友誼就如酒，永遠(yuǎn)都會久，重陽登高日，祝福送給你，愿你重陽佳節(jié)日，開心無限期!九九重陽節(jié)祝福語

3.歲歲重陽，今又重陽，傲霜滿地香;久久思量，久久難忘，九九重陽勝春光;美酒一杯，秋雁兩行，福澤深厚又綿長。祝重陽快樂，永遠(yuǎn)健康!

4.歲歲重陽，今又重陽，九九相逢祥瑞降。不似春光，勝似春光，秋高氣爽喜洋洋。金菊飄香，歸雁成行，健康喜樂福綿長。重陽節(jié)，祝你幸福安康!

5.松柏不殘四季翠，山村難老百歲人，三三令節(jié)春時(shí)松，更高九九芳辰重陽鶴添壽，愿秋風(fēng)捎去我的思念和祝福，祝你越活越精神，越活越年輕!

6.思念在歲月中蕩漾，南飛的大雁披上了霓裳，金菊在清風(fēng)中綻放，轉(zhuǎn)眼又到了重陽?？匆换靥旄咴频?，走一路山高水長，道一聲幸福安康。重陽快樂!

7.說重陽，道重陽，收獲季節(jié)，遍地金黃;秋風(fēng)吹動，秋高氣爽，稻子熟透，漸香，歲月歲歲重陽，今又重陽，不是春光，勝似春光，一根扁擔(dān)，兩挑籮筐，三秋雁陣，四分五行，六六大順，七下八上，九九重陽，十月農(nóng)忙，短信在此，祝你健康!

8.雖然我不能陪你品酒賞菊，但我們可以共賞同一條短信;雖然我不能陪你登高插萸，但我們可以轉(zhuǎn)發(fā)同一條祝福;重陽節(jié)，總有一條短信專門送給你。

9.送一縷陽光給你，在秋日的蕭瑟中為你捎去溫暖，送一份祝福給你，在重陽佳節(jié)即將到來的日子里為你帶來關(guān)懷，預(yù)祝重陽節(jié)快樂，天天快樂。

10.送上一支山茱萸，愿你永遠(yuǎn)愉快;送上一塊重陽糕，愿你永遠(yuǎn)高興;送上一杯酒，愿你幸福久久。重陽節(jié)到了，祝福你節(jié)日天天好心情!

11.送你清風(fēng)讓你心曠神怡，送你細(xì)雨洗去你疲憊的汗跡，讓藍(lán)天作封白云作信，讓流星作我的特快專遞，送你彩虹通向夢想之旅，祝你重陽節(jié)快樂!

12.如歌，天地變涼，思念不變，祝福奉上。重陽節(jié)快樂!

13.霜降遇重陽送你養(yǎng)身小方子，沒事洗個(gè)鴨梨子，潤潤小嗓子，睡覺蓋好了被子，趕走疾病的引子，打著問候的旗子，送來好運(yùn)的影子，祝你快樂過日子!

14.雙九重陽佳節(jié)到，短信祝福來問好：相逢總是很奇妙，生活無憂無煩惱，工作壓力瞬間消，傷心失落全跑掉，鍛煉身體多散步，愿你重陽更美好!2013重陽節(jié)祝詞

15.雙九節(jié)，出游賞景好時(shí)節(jié);九九節(jié)，登高遠(yuǎn)眺值佳節(jié);重陽節(jié)，觀賞好季節(jié);遍插茱萸、吃重陽糕、飲酒，可去邪，祝君重陽過好節(jié)!

16.帥哥味好純：但得夕陽無限好，何須惆悵近黃昏，愿你擁有一個(gè)美好、快樂的節(jié)日!秋風(fēng)徐徐，重陽九九，蒸上九重的粟糕，備好香醇的菊酒，等著與你分享。

17.歲歲重陽，今又重陽，又見香;不是春光，勝似春光，秋色迷人心芬芳。讓我登上高處，為你送來真誠和愛的祝福：祝重陽快樂，合家康泰!

18.歲歲重陽，今又重陽，誰道秋日涼?重陽節(jié)里歌聲放，祝福飛來心飛揚(yáng)。九月九里共舉杯，聲聲祝福暖心扉。短信送到你這里，祝福也能化傷悲!

歲歲又重陽范文第4篇

2、我輕呵出一團(tuán)氣，將滿天飄舞的思緒，凝結(jié)成一朵白云，用心把它繪的五彩斑斕，載上沉淀的祝福，帶去來自遠(yuǎn)方的重陽節(jié)的問候——送給永遠(yuǎn)開心的你!

3、歲歲重陽，今又重陽，不似春光，勝似春光。松柏不殘四季翠，山村難老百歲人。祝您重陽節(jié)快樂!

4、佛說：重陽節(jié)又到，要相約登高，要常思故友，尤其是現(xiàn)在給你發(fā)短信的人!要經(jīng)常請他吃飯!特別是九月九!要把你最最真誠的祝福都給他!善哉!善哉!

5、回鄉(xiāng)的路，曲曲彎彎，轍痕深淺?；丶业男模缭缤硗?，沖過籬藩。故園的情，歲歲年年，月掛云端。又到重陽倍思親，異鄉(xiāng)客，心相近，祝吾友，多溫馨!

6、

7、愿友情像秋日的紅葉那樣美麗，愿你心情像秋日的藍(lán)天那樣爽朗，愿愛情像秋日的果實(shí)一樣豐碩，愿老朋友重陽節(jié)開心快樂!

8、一九二九，天長地久;三九四九，幸福永久;五九六九，福祿長久;七九八九，神交已久;重陽九九，相伴久久!重陽節(jié)快樂!

9、九九重陽，因?yàn)榕c“久久”同音，九在數(shù)字中又是數(shù)，有長久長壽的含意，況且秋季也是一年收獲的黃金季節(jié)，重陽佳節(jié)，寓意深遠(yuǎn)。祝你重陽節(jié)快樂!

10、重陽節(jié)到，我向月老許愿讓你幸福，他答應(yīng);我向財(cái)神許愿讓你富裕，他點(diǎn)頭;我向王母許愿。。。玉帝說：諸神放假過節(jié)，干脆祝收到短信的人心想事成吧!

11、您生命的秋天，是楓葉一般的色彩，不是春光勝似春光，時(shí)值霜天季節(jié)，卻格外顯得神采奕奕。祝您老重陽節(jié)快樂，健康長壽!

12、夜里開機(jī)看見，重陽佳節(jié)祝福。千萬里送來問候，萬千言語祝福，溫暖你心扉。我的短信飛快，猶如光速傳載，了卻重陽開心事，贏得佳人美酒歸，歌聲滿天飛!

13、重陽佳節(jié)到來，親朋好友開懷，祝你心情愉快，心事統(tǒng)統(tǒng)放開。九月九，讓我們一起登高望遠(yuǎn)吧，借著醇香的美酒，分享心中的憂愁與快樂，重陽節(jié)快樂!

14、九月九，重陽節(jié)。梳個(gè)鬏，換個(gè)裝。拜個(gè)舅，唱個(gè)喏。喝個(gè)酒，散個(gè)心，敘個(gè)舊，念個(gè)情。好日子，年年過。重陽節(jié)，更快樂。

15、又是一年重陽時(shí)，花點(diǎn)小錢祝福你;秋高氣爽好時(shí)節(jié)，登高賞菊最適宜;發(fā)封短信問候你，愿你事事都順利!祝重陽節(jié)快樂!

16、九九重陽節(jié)，九個(gè)祝福送給你，一祝身體好，二祝心情妙，三?？鞓防@，四祝好運(yùn)到，五祝來鈔票，六祝煩惱跑，七祝永不老，八祝萬事好，九祝短信告。

17、生活貧窮還是富裕，開心就好;工作辛苦還是輕松，快樂就好;朋友近鄰還是遠(yuǎn)離，惦記就好;祝福簡短還是冗長，真心就好!祝你九九重陽快樂一切都好!

18、千重坎萬重關(guān)，愿快樂陽光伴你燦爛;千重水萬重山，愿好運(yùn)陽光陪在你身邊;千重地萬重天，愿成功陽光溫暖你心間。重陽佳節(jié)，愿久久好運(yùn)把你圍繞!

19、九九重陽節(jié)，老人最希望健康永久，孩子最盼望快樂長久，情人最期望天長地久，上班族最渴望活力持久，在這個(gè)悠久的節(jié)日，我最想說：祝你好運(yùn)更長久!

20、當(dāng)國慶遇到重陽，這是盛世和傳統(tǒng)的碰撞：是要清楚告訴我們，出游歡樂的時(shí)光，莫忘家中蒼老的爹娘。陪父母登高望遠(yuǎn)賞菊香，祝愿父母快樂安康，莫忘!

21、九九重陽兩九相遇，物極必反，萬物凋零。我的祝福經(jīng)年累月伴隨你，不會斷絕，不會凋零，永遠(yuǎn)年輕。重陽節(jié)祝你帶著快樂迎接秋雨，帶著溫暖迎接嚴(yán)寒。

22、重陽氣壞了灰太狼，不能團(tuán)聚還要抓羊。對著發(fā)毒誓，一定殺盡小綿羊。他正趕著去找你，你要小心的提放。重陽節(jié)里多快樂，你這可愛的小綿羊。

23、飲一碗美酒，登一座高山，插一片茱萸，賞一壇，掬一把相思，送一份祝福，又逢重陽佳節(jié)，我祝您節(jié)日快樂，健康幸福!

24、一年一度重陽，祝福大不一樣，生活喜氣“陽陽”，職場“陽陽”得意，幸福身邊徜“陽”，愛情沐浴陽光，心情陽光燦爛。收下祝福吧!重陽節(jié)快樂!

25、重陽節(jié)到了，翻遍《辭?！芬矝]有找到一句值得送給你的祝福，所以只能俗氣地祝你享的福氣如東海泛濫，你領(lǐng)的薪水如南海澎湃。重陽“海皮”!

26、我用彩云編織繾綣的夢境，收集心中每一份感動，許下星空每一個(gè)祝愿，記載心頭每一絲企盼，交織成一首幸福美麗的樂章，在這重陽佳節(jié)伴隨你!

27、秋來天氣漸轉(zhuǎn)涼，不知不覺到重陽。溫馨問候情意長，真誠關(guān)懷存心房。秋菊艷艷多欣賞，秋雨瀟瀟添衣裳。秋風(fēng)送去我祝福，幸福永伴你身旁。重陽快樂!

28、歲歲重陽，今又重陽，傲霜滿地香;久久思量，久久難忘，九九重陽勝春光;美酒一杯，秋雁兩行，福澤深厚又綿長。祝重陽快樂，永遠(yuǎn)健康!

歲歲又重陽范文第5篇

文案一

1、送你一棵茱萸，愿你大吉大利；敬一杯酒，祝你健康又長壽；送你一架云梯，助你登高遠(yuǎn)望；送你一句祝福，重陽節(jié)快樂！

2、有些往事，即使遠(yuǎn)去也會在心頭縈繞；有些人，即使不常聯(lián)系也會常記在心里；就像你，只輕輕的一個(gè)短信，就是我衷心心意的快遞！祝：重陽快樂！

3、藍(lán)天作信封，白云作信紙，流星是偶的特快專遞。送你清風(fēng)讓你心曠神怡，送你細(xì)雨洗去你疲憊的汗跡，送你彩虹通向夢想之旅重陽節(jié)！重陽節(jié)快樂！

4、九月初九重陽節(jié)，遵守傳統(tǒng)好風(fēng)俗。登高遠(yuǎn)眺金秋景，寓意吉祥步步高。飲酒祈福賞，保佑壽比南山松。尊俗重道度重陽，祝你幸福久久長。

5、朵朵迎風(fēng)開，悠悠花香沁心脾；九九重陽翩翩至，喜上心頭笑開懷；漫步郊外心情爽，徒步登高樂逍遙；舉目遠(yuǎn)眺觀美景，心曠神怡爽歪歪；朋友特來送祝福，情誼綿綿深又厚；祝你重陽好心情，生活幸福喜滋滋！

6、朵朵迎風(fēng)開，悠悠花香沁心脾；九九重陽翩翩至，喜上心頭笑開懷；漫步郊外心情爽，徒步登高樂逍遙；舉目遠(yuǎn)眺觀美景，心曠神怡爽歪歪；朋友特來送祝福，情意綿綿深又厚；祝你重陽好心情，生活幸福喜滋滋！

7、九九重陽是中國文化中最吉祥的日子，在這金風(fēng)送爽，桂花飄香，爭艷的日子里，我祝各位幸福久久，健康久久，更祝長輩們多福多壽。祝重陽節(jié)快樂！

8、送你一盒重陽糕，它以祝福與問候?yàn)樵?，相間疊放：一層幸福粉，一層平安糖，一層開心豆，一層健康果，愿你重陽節(jié)快樂。

9、藍(lán)天作信皮，白云作信紙，流星是我的特快專遞。送你清風(fēng)讓你心曠神怡，送你細(xì)雨洗去你疲憊的汗跡，送你彩虹通向夢想之旅，祝你重陽節(jié)快樂！

10、重慶遇見重陽就是雙重，今天祝福與問候雙重，祝你工作與生活雙重棒，健康與快樂雙重好，幸福與甜蜜雙重久，此短信你如果收到兩遍，那是因?yàn)槲野l(fā)重了。祝重陽節(jié)快樂！

11、遍地的茱萸都想戴在你頭上，長出很多的幸福吉祥，我們一起歡暢瘋狂；登到最高處哪怕是你的心上，讓金秋的為你飄香，九月九我們的重陽，祝你生活步步高，幸運(yùn)又安康。

12、月九，送美酒，幸福酒，好運(yùn)旺，快樂人生樂安康；友誼酒，情意深，久久的祝福長又長；九月九，重陽到，祝你快樂久久！

13、輕輕打開你的手機(jī)，慢慢翻看我的祝福：愿今天的你：快樂！今年的你：順利！今生的你！健康！今世的你：幸福！恭祝您及家人重陽快樂幸福！

14、金秋時(shí)節(jié)到，空氣好又好，九九重陽節(jié)，悄悄來到了，登高插茱萸，這個(gè)少不了，暢飲酒，這個(gè)最為妙，發(fā)個(gè)短信息，這個(gè)最重要，身體要健康，這個(gè)我看好。

15、可能你不知道重陽為什么要登高，不知道重陽為什么要插茱萸，不知道重陽為什么要賞，但是我知道發(fā)這條短信是為了給你帶去真切的祝福！重陽節(jié)快樂！

16、我在手機(jī)的這邊，你在手機(jī)的那邊，中間是無形的電波，距離阻隔了相見，卻阻隔不了祝福，讓這條短信隨著無形的電波傳送到你的手機(jī)中，重陽節(jié)快樂！

17、九九重陽，好事成雙，送給你雙重的祝福：快樂+快樂=快快樂樂，圓滿+圓滿=圓圓滿滿，長久+長久=長長久久。祝你快快樂樂圓圓滿滿長長久久。

18、重陽節(jié)快樂，一句最簡單的祝福送給你，希望你天天都能健康快樂，忘記心中的.煩悶，擁有一份陽光般燦爛的心情。

19、在思念的日子里，留下了如同秋葉般數(shù)不盡的美好回憶，我托秋雁把最真心的祝福送給你：愿你在重陽節(jié)里美麗如金秋菊，好運(yùn)如重陽糕，甜美如重陽酒。

20、九月九日上酒樓，酒醉醉酒忘憂愁，久久凝思九九日，九九重陽想舅舅，糾結(jié)思念家中舅，久久未見離家久，帶酒一瓶送舅舅，祝福永久在心中。

21、重陽節(jié)到了，祝你登高望遠(yuǎn)，事運(yùn)，財(cái)運(yùn)，愛情運(yùn)，運(yùn)運(yùn)皆高，祝你把酒暢歡，人歡，心歡，情歡，事事皆歡！重陽節(jié)快樂！

22、處事秘笈：做人要像李莫愁，刮風(fēng)下雨都不愁；做事要像張無忌，牛鬼蛇神都不懼；工作要像王重陽，全真七子來幫忙；生活要像韋小寶，快樂幸福沒煩惱！

23、稻花香，插茱萸，登高望遠(yuǎn)喝小酒。吃花糕，又相聚，賞菊品茗會摯友。九月九，重陽日，健康開懷人長久。重陽節(jié)即將到來，短信預(yù)祝重陽快樂！

24、九九重陽，沉淀著金秋的豐碩，感受了生活的溫馨。它是屬于花甲卸任轉(zhuǎn)折的輕松、古稀牽手連接的和睦、耄耋拐杖支撐的充實(shí)的。一句貼心的問候、一條溫馨的短信，祝愿所有朋友節(jié)日快樂。

25、重陽節(jié)到了，看在我這個(gè)這么"重"量級人物在這個(gè)"陽"光燦爛的節(jié)日里給你祝福問候的份上，你怎么得也該高興快樂吧！重陽節(jié)快樂哦！

26、正值秋高氣爽，人壽花香，一條短信帶來我的九九的渴望；正值彩云追月，桂花飄香，一條短信帶來我的九九的衷腸，祝你開心重陽，幸福重陽！

27、重陽節(jié)，更是老人的節(jié)日，您生命的秋天，是楓葉一般的色彩，不是春光勝似春光，霜天季節(jié)，卻格外神采奕奕。祝老人們重陽節(jié)快樂，健康長壽！

28、邀你一起品嘗九月九的酒，能賞個(gè)臉嗎？重陽節(jié)快樂！

29、九九重陽節(jié)就要到了，提前祝愿所有活躍的、沉默的、熱情的、深沉的、玩酷的、耍賴的、在線的、隱身的朋友們：重陽節(jié)快樂！

30、輕風(fēng)拂山崗，遍野黃，佳節(jié)到重陽，思君登高望。情意心中藏，祝福暖心房，快樂似水長，好運(yùn)伴君旁。鴻雁帶福至，悠閑莫匆忙，天高云淡時(shí)，愿你永安康！

31、今又重陽，祝福送上，敬老踏秋，只為安康。登高眺望，前程光芒，輕嗅菊香，心情舒暢，糕點(diǎn)黃黃，幸福蕩漾，九九節(jié)日，祝你全家快樂流長。

32、為了讓你更加開心，為了讓你忘記鉸單，為了讓你感受到生活的美好，請大聲地念出一下的祝福語：重陽節(jié)快樂。

33、酒是什么樣的酒，友情長青酒；久是什么樣的久，天長又地久；九是什么樣的九，一四六八九，祝你豐"一"足"四"，工作順"六""八"大財(cái)。重陽快樂！

34、獨(dú)在異鄉(xiāng)過重陽，每逢佳節(jié)把你想，不知好友怎么樣，編條短信送吉祥。過個(gè)重陽不容易，即使不能在一起，祝福收到笑嘻嘻：祝重陽節(jié)快樂重重！重陽節(jié)快樂！

35、雙九重陽佳節(jié)到，微信祝福來問好：相逢總是很奇妙，生活無憂無煩惱，工作壓力瞬間消，傷心失落全跑掉，鍛煉身體多散步，愿你重陽更美好！

36、九月九，送美酒，桃花酒，姻緣到，甜蜜愛情香又香；幸福酒，好運(yùn)旺，快樂人生樂安康；友誼酒，情意深，久久的祝福長又長；九月九，重陽到，祝你快樂久久，"久"是幸福喲！

37、恁個(gè)久沒盯到你，重陽節(jié)我還是整條短信，假巴意思問候哈你嘛！祝愿你身體巴巴適適勒，生活得安安逸逸勒，工作得耍耍搭搭勒，日子過得撐撐抖抖勒！

38、您生命的秋天，是楓葉一般的色彩，不是春光勝似春光，時(shí)值霜天季節(jié)，卻格外顯得神采奕奕。祝您老重陽節(jié)健康長壽！

39、但是夕陽無限好，何須惆悵近黃昏！ 蒼龍日暮還行雨，老樹春深更著花！ 祝您越活越精神！年年重陽都快樂！

40、曾經(jīng)您告訴我，三月三風(fēng)箏飛上天，那是童年的快樂?，F(xiàn)在，九月九香，是老人節(jié)，是您的節(jié)日，親愛的爹娘，祝你們身體安康、福壽永享，重陽節(jié)快樂！

41、想念你，在每一個(gè)嫁接；祝福你，在每一個(gè)新的一天；愿快樂在你身邊圍繞，讓美夢總是在你身邊守候。重陽來臨的時(shí)刻，愿你今天比昨天更快樂！

42、斟上一杯重陽酒，遙表思念牽掛長，身插茱萸祈平安，悠悠惦念在心間，又到一年重陽節(jié)，點(diǎn)點(diǎn)祝福隨風(fēng)傳，重陽節(jié)，愿你平安幸福，重陽快樂！

43、愿你薪水多賺，獎金滿滿；快樂天天，幸福年年！祝我們這些表面風(fēng)光內(nèi)心彷徨；容顏未老，心已滄桑；成就難有，郁悶經(jīng)常；比螞蟻忙；比岳飛更忠良的兄弟姐妹們，重陽節(jié)快樂！

44、時(shí)光流逝，流不走我心中的你！將萬千思念托付那吉祥鳥，帶去我深深的問候；時(shí)間阻擋不了那份執(zhí)著，不管未來之路多么崎嶇，今天給你送上重陽祝福！祝你重陽節(jié)快樂！

45、一絲秋雨秋意濃，一縷秋風(fēng)秋情動，一輪明月誰與共，一份思念遙相送，一枝茱萸情義重，一朵香入夢，一條微信你會懂，一片真情重陽祝福中。

46、心存明月，月照九州，可知人生本無常？正香，今夕昨昔相聚相離，琴瑟琵琶絲竹管弦，遍插茱萸，花中行樂月中眠，半醉半醒，譜寫重陽喜慶的樂章

47、凋零的不是那季節(jié)，而是花朵；成熟的不是那果實(shí)，而是歲月；南飛的不是那大雁，而是心情；牽掛的不是那日子，而是家人。重陽節(jié)，氣候變，請您注意健康！

48、九月九，重陽節(jié)，一樣的季節(jié)，一樣的日子，一樣的牽掛，一樣的思念，一樣的深情，一樣的心情，一樣的問候，一樣的祝福，愿你擁有一個(gè)愉快的重陽佳節(jié)！

49、重陽節(jié)到了，回憶起你我當(dāng)初在一起的日子，那是一段永遠(yuǎn)難忘的友情歲月，當(dāng)你在孤獨(dú)時(shí)別忘記還有一位老朋友惦記著你，真心祝你一切順利，天天開心。

50、歲歲重陽，今又重陽，九九相逢祥瑞降。不似春光，勝似春光，秋高氣爽喜洋洋。金菊飄香，歸雁成行，健康喜樂福綿長。重陽節(jié)，祝你幸福安康，和和美美！

文案二

1、秋風(fēng)涼，秋菊香，秋景中迎來重陽，合家歡，兄弟聚，好日子里秉燭敘，插茱萸，齊登高，九九快樂更美好，思念深，祝福來，祝你重陽開心愉快！

2、重陽佳節(jié)憶兄弟，千言萬語不足惜；有人登高去遠(yuǎn)眺，有人林中來采菊；我在這里想著你，愿你重陽好運(yùn)氣；幸福全都在心底，健康好夢更吉利！

3、重陽佳節(jié)團(tuán)聚，親朋好友團(tuán)聚，情人戀人團(tuán)聚，你我他團(tuán)聚，在這個(gè)快樂的節(jié)日里！真心祝福你：事業(yè)豐收，薪水豐收，愛情豐收，快樂永相伴！

4、重陽佳節(jié)珍貴，周末不能浪費(fèi)。出門旅游太累，登高望遠(yuǎn)太貴?；丶倚⒕锤改福硎苡H情安慰。嘗嘗媽媽手藝，美食值得回味。家庭和諧幸福，工作才不疲憊！

5、重陽佳節(jié)又來臨，送你幾條小建議，出游一定不要去，費(fèi)錢費(fèi)神又費(fèi)力，登高危險(xiǎn)系數(shù)大，一不小心就要上擔(dān)架，不如發(fā)個(gè)短信，打個(gè)電話，陪我一起訴牽掛！

6、重陽將來到，快樂步步高。人間重晚晴，殷殷寸草心。霜葉紅勝花，歲歲好年華。莫道桑榆晚，幸福金不換。愿你重陽享安康，吉祥伴身旁。

7、重陽佳節(jié)來相聚，親朋好友在一起，情人戀人笑歡語，你我短信傳祝福。讓我們?yōu)槊魈烀篮玫那俺滔嗑郯桑∠嗑壑仃?，相守未來，重陽快樂?/p>

8、莫道重陽秋風(fēng)涼，果實(shí)累累喜洋洋；莫道重陽雨霏霏，紅葉掛滿心頭醉；莫道重陽夕陽紅，閱盡風(fēng)霜霞光美；莫道重陽話凄涼，登高遠(yuǎn)眺思緒飛！祝重陽節(jié)快樂！

9、飄香，秋風(fēng)清涼，高望遠(yuǎn)，開心舒暢，遍插茱萸，互祝健康，重陽佳節(jié)，兄弟同堂，故人重逢，暢抒心房，親情友情，隨秋高揚(yáng)，共度重陽，同慶安康。

10、重陽佳節(jié)到來，愿你心里幸福暖洋洋，開心笑聲隨風(fēng)揚(yáng)，美好心情任傳揚(yáng)，夫妻恩愛似鴛鴦，友情愛情齊蕩漾，事業(yè)身體皆無恙，喜慶多多勝驕陽。

11、重陽佳節(jié)送你一條充滿法力的短信，收到的人將會時(shí)來運(yùn)轉(zhuǎn)；前途平坦，兜里有款，甜長苦短，薪水翻番，好吃好穿，常有新歡，追求的路程越走越寬。

12、重陽佳節(jié)送你限量版重陽糕，祝你心情好開心程度高，工作順辦事效率高，薪水漲升級職位高，身體棒生活質(zhì)量高，朋友多人氣指數(shù)高，福氣到運(yùn)氣與天齊高！

13、重陽佳節(jié)送你幾顆心：早上出門舒心！路上小心！遇事耐心！做事細(xì)心！交友留心！待人誠心！對自己有信心！對情人有愛心！更重要的是重陽佳節(jié)你最開心！

14、重陽將來到，快樂步步高。人間重晚晴，殷殷寸草心。霜葉紅勝花，歲歲好年華。莫道桑榆晚，幸福金不換。愿你重陽享安康，吉祥伴身旁。

15、歲歲重陽，今又重陽，九九相逢祥瑞降。不似春光，勝似春光，秋高氣爽喜洋洋。金菊飄香，歸雁成行，健康喜樂福綿長。重陽節(jié)，祝你幸福安康！

16、我愿化作一片云，隨你漂流到天涯海角，我愿變成一首歌，寂寞時(shí)在你耳邊響起，我用心編寫最真摯的祝福，在這重陽佳節(jié)里送給您，愿您越活越精彩！

17、重陽佳節(jié)九月九，贈你酒，祝你平安健康久；邀你爬山觀秋景，登高轉(zhuǎn)運(yùn)好運(yùn)久；與你同享蜜花糕，事業(yè)甜蜜快樂久；送你一方茱萸佩，趨吉迎祥幸福久。

18、重陽佳節(jié)九月九，美好祝福送不夠。生活甜蜜如美酒，愛情幸福到永久。工作輕松常袖手，家庭和諧福常有。諸事順利煩惱走，一切如意喜臨頭。

19、思念如一葉扁舟，在歲月的長河中漂流，劃過一年一年一天一天。記錄點(diǎn)點(diǎn)滴滴，絲絲縷縷，到重陽節(jié)那天，一觸即發(fā)！祝重陽節(jié)快樂！

20、九月九，再聚首，重陽相聚會老友；是親友，是朋友，一切盡在杯中酒；情也深，愛也久，佳節(jié)團(tuán)圓來敘舊；念今昔，盼永久，每年相聚九月九。重陽節(jié)快樂！

21、送你一盒重陽糕，它以祝福與問候?yàn)樵?，相間疊放：一層幸福粉，一層平安糖，一層開心豆，一層健康果，愿你重陽節(jié)快樂。

22、重陽臨，提前許九個(gè)美麗愿望送你：一愿心情好，二愿憂愁拋，三愿幸福繞，四愿收入高，五愿身體康，六愿煩惱消，七愿不變老，八愿困難少，九愿樂逍遙！

23、重陽佳節(jié)九月九，吉祥好運(yùn)送朋友：事事順心好運(yùn)有，煩惱沒有幸福留；親情友情和愛情，猶如日月天長久；短信祝福送你手，美滿如意到久久。重陽節(jié)快樂！

24、秋菊，傾吐悠悠清香；茱萸，環(huán)繞片片吉祥；登高，遙寄無限思念；酒杯，斟滿情誼美酒；心靈，發(fā)送誠摯祝福：九九重陽，愿你開心，心情愉悅！

25、將彩云編織繾綣的夢境，收集心中每一份感動，許下星空每一個(gè)祝愿，記載心頭每一絲企盼，交織成一首幸福美麗的樂章，在這重陽佳節(jié)伴隨你！

26、祝福你理想幻想夢想心想事成；公事私事心事事事稱心；財(cái)路運(yùn)路人生路路路暢通；晴天雪天天天開心，親情友情愛情情情似海；重陽節(jié)快樂！

27、重陽佳節(jié)秋高氣爽，登高鍛煉飄香，遙望遠(yuǎn)方思緒飛揚(yáng)，家中親人是否安好？發(fā)條短信道聲平安，問聲安好表表心愿，祝福親友久久好運(yùn)久久健康！

28、重陽佳節(jié)秋風(fēng)涼，多添衣來保安康，時(shí)節(jié)好做茶，瀉火安神心清爽，天漸干多多喝水，夜?jié)u長作息如常，飲食注意規(guī)律，鍛煉適當(dāng)加強(qiáng)，愿重陽再添安康。

29、有你相伴，黃金不換，有我祝福，身體舒服，快樂重陽，天天吉祥，短信字少，情意卻多，在這美好的節(jié)日里祝你永遠(yuǎn)幸福，重陽節(jié)快樂！

30、重陽節(jié)，登高行。鮮花旁，笑臉映?？諝饫剩榍屣L(fēng)。吉祥茶，如意餅。邀親朋，相思情。若懷念，望熒屏。問候語，表心聲。遙祝愿，笑不停。祝重陽快樂！

31、歲歲重陽，今又重陽，又見香；不是春光，勝似春光，秋色迷人心芬芳。讓我登上高處，為你送來真誠和愛的祝福：祝重陽快樂，合家康泰！

32、歲歲重陽，今又重陽，誰道秋日涼？重陽節(jié)里歌聲放，祝福飛來心飛揚(yáng)。九月九里共舉杯，聲聲祝福暖心扉。短信送到你這里，祝福也能化傷悲！

33、重陽節(jié)到了，也沒啥好送你的，就送你一束吧。可別小看它，你可以用它釀成酒，飲后無憂愁；還可制成糕，吃后樂逍遙。重陽節(jié)快樂！

34、歲歲重陽，今又重陽，不是春光，勝似春光，一根扁擔(dān)，兩挑籮筐，三秋雁陣，四分五行，六六大順，七下八上，九九重陽，十月農(nóng)忙，短信在此，祝你健康！

35、雖然我不能陪你品酒賞菊，但我們可以共賞同一條短信；雖然我不能陪你登高插萸，但我們可以轉(zhuǎn)發(fā)同一條祝福；重陽節(jié)，總有一條短信專門送給你。

36、重陽佳節(jié)翩然到，尊長敬老盡孝道，登高祈福好運(yùn)臨，家人同食重陽糕。金黃富貴臨，茱萸懸門福氣繞，踏秋田野賞美景，舉杯美酒吉祥兆。愿你重陽心情好，家人團(tuán)聚樂逍遙，健康常在迎祥瑞，福氣臨門好運(yùn)到！

37、重陽佳節(jié)到來，親朋好友開懷，祝你心情愉快，心事統(tǒng)統(tǒng)放開。九月九，讓我們一起登高望遠(yuǎn)吧，借著醇香的美酒，分享心中的憂愁與快樂，重陽節(jié)快樂！

38、重陽佳節(jié)到來了，親朋好友開懷笑，心情愉快外出游，心事統(tǒng)統(tǒng)來丟掉。九月初九好日子，登高望遠(yuǎn)吉祥擁，醇香美酒舉杯飲，分享心中憂與樂。祝愿朋友重陽節(jié)快樂！

39、空氣中彌漫著歡樂，樹梢上飄落著祝福，重陽的溫馨在招手，節(jié)日的激情在噴薄。偶愿化作清風(fēng)、陽光、白云，給你載來如意、健康、財(cái)富。重陽節(jié)快樂重陽節(jié)來歷！

40、帶一顆心去登高，讓一雙眼去望遠(yuǎn)，攜一份情懷去思念，寄一縷清風(fēng)去相見，托一個(gè)好夢去連線，送一聲祝福去你身邊。重陽節(jié)，愿你步步高，天天好。

41、送一縷陽光給你，在秋日的蕭瑟中為你捎去溫暖，送一份祝福給你，在重陽佳節(jié)即將到來的日子里為你帶來關(guān)懷，預(yù)祝重陽節(jié)快樂，天天快樂。

42、送上一支山茱萸，愿你永遠(yuǎn)愉快；送上一塊重陽糕，愿你永遠(yuǎn)高興；送上一杯酒，愿你幸福久久。重陽節(jié)到了，祝福你節(jié)日天天好心情！

43、送你清風(fēng)讓你心曠神怡，送你細(xì)雨洗去你疲憊的汗跡，讓藍(lán)天作封白云作信，讓流星作我的特快專遞，送你彩虹通向夢想之旅，祝你重陽節(jié)快樂！

44、重陽敬請歇一歇，又是一個(gè)傳統(tǒng)節(jié)；南方飄雨北方雪，朋友還是我倆鐵；人生相逢皆緣起，日月輪轉(zhuǎn)點(diǎn)點(diǎn)滴；祝福就是送給你，祝你快樂又如意！

45、九九芳辰重陽鶴添壽，愿秋風(fēng)捎去我的思念和祝福，祝你越活越精神，越活越年輕！

46、借此佳節(jié)之際我感謝你，你的笑顏似燦爛的陽光照亮了我的'世界，你給予我的安慰和鼓勵(lì)支持我度過一切艱苦，言語不能表達(dá)我對你的情誼，你是一位了不起的人。祝你重陽節(jié)快樂！

47、江涵秋影雁初飛，與客攜壺上翠微。塵世難逢開口笑，須插滿頭歸。但將酩酊酬佳節(jié)，不作登臨恨落暉。古往今來只如此，牛山何必獨(dú)沾衣。

48、紅顏未虛伏虎降龍，白發(fā)帶笑鋪金疊翠！天高氣爽，人壽花香！