接种后的

The cells labeled with PKH26 were seeded on the bio-derived bone to construct tissue engineered bone in vitro. Then the compound of cells and material were observed under fluorescence microscope. The compound of labeled cells and material were implanted into the rabbit thigh muscle, and the transformation of the labeled cells was observed by fluorescence microscope 14 and 28 days later.

将PKH26 标记的BMSCs 接种于生物衍生骨材料，48 h 后荧光显微镜观察细胞与材料的复合情况；将细胞-材料复合物植入成年新西兰大白兔双后肢肌袋，14、28 d 后荧光显微镜观察标记细胞的转归。

To explore the possibility of early and rapid diagnosis of mycosisby detecting the fungal autofluorescence. Methods: To culture M. furfur and M. japonica standard strains on Dixonmedium at 32℃, and to study the autofluorescence of colonies in 2nd, 4th, 6th, 8th, 10th day under Laser Scanning Confocal Microscopy (LeicaTCS-SP2 LSCM). To study the variation of autofluorescence whenMalassezia colonies were put in 3% glutaraldehyde, pH4.0 HCL, 10% KOH, fluconazol solution, 0.3% methylene blue separately. To detect theautofluorescence of C. albicans, S. schenchii, T. rubrum, T. tonsurans, A.terreus, A. fumigatus cultured on Sabouraud dextrose agar.

选用糠秕马拉色菌和日本马拉色菌2株标准菌，接种在Dixon培养基32℃培养，分别在培养的第2、4、6、8、10天挑取菌落放在TCS-SP2型激光扫描共聚焦显微镜(Leica TCS-SP2 LSCM)下观察、测定其自发荧光；分别用3％戊二醛溶液、pH4.0盐酸溶液、10％氢氧化钾溶液、1280μg／mL和64μg／mL氟康唑溶液、0.3％美蓝溶液处理后观察马拉色菌自发荧光的变化；选择白念珠菌、申克孢子丝菌、红色毛癣菌、断发毛癣菌、土曲霉、烟曲霉等用沙堡葡萄糖琼脂培养后观察其自发荧光的特点。

Cubense, indicating that the expression of M-PAL may be related to resistance of banana Fusarium wilt.

cubense race 4 后大蕉叶片中M-PAL基因的转录谱进行研究表明，在接种枯萎病菌后M-PAL基因在叶片中的转录水平提高，因此推测M-PAL基因的表达可能与香蕉枯萎病抗性相关。

Sp. cubense, indicating that the expression of M-PAL may be related to resistance of banana Fusarium wilt.

sp。 cubense race 4后大蕉叶片中M-PAL基因的转录谱进行研究表明，在接种枯萎病菌后，M-PAL基因在叶片中的转录水平提高，因此推测M-PAL基因的表达可能与香蕉枯萎病抗性相关。

METHODS: Gastric carcinoma cell line MKN45 at logarithmic growth phase was subcutaneously inoculated in 20 athymic mice.

将对数生长期的胃癌细胞株MKN45接种到裸鼠皮下，成瘤后用制备好的分别含GCRG213正、反义克隆和RNAi片段的腺相关病毒分别在肿瘤局部注射，以空病毒和生理盐水对照，继续喂养2 wk后处死，取肿瘤测量重量，并用半定量RT-PCR检测肿瘤组织GCRG213mRNA表达水平。

METHODS: 10 mL bone marrow was extracted from dog posterior superior iliac spine, heparin anticoagulation, diluted using Hanks juice, treated with 1.077 g/mL Ficoll solution 3 mL, and centrifuged at 2 000 r/min for 20 minutes. Karyocytes were absorbed to form white cloudlike layering interface, and then centrifuged twice using DMEM supplemented with fetal bovine serum, incubated at 12×104/cm2 at 37 ℃ in a 5% CO2 incubator.

于犬髂后上棘抽取骨髓液10 mL，肝素抗凝，Hanks液稀释后，加入1.077 g/mL Ficoll液3 mL，2 000 r/min离心20 min，吸取有核细胞形成白色云雾状的分层界面，用含胎牛血清的DMEM离心2遍，按12×104/cm2密度接种，于37 ℃、体积分数为5%的CO2培养箱内培养。

Alloy of polyethylene of element of the methodological freeboard that make, titanium and sample of alloy of cobaltic chromic molybdenum, FITC of classics of cuticular grape coccus labels, the cuticular grape coccus that after sample of artificial and articulatory material is disinfected, vaccinal FITC labels, sample surface cent is slick surface and rough surface, every groups each 6 sample, brood the 24 orifice plate that contain bacterium and sample below 37 ℃ after Yo 30min, with fluorescent microscope observation, sample uses observation of scanning report lens.

方法制作超高分子聚乙烯、钛合金和钴铬钼合金试样，表皮葡萄球菌经FITC标记，人工关节材料试样消毒后接种FITC标记的表皮葡萄球菌，试样表面分为光滑表面和粗糙表面，每组各6个试样，将含有细菌和试样的24孔板在37℃下孵育30min后，用荧光显微镜观察，试样用扫描电镜观察。

The introduction combines traditional Chinese and Western medicine below the experience of enteritis of noxiousness of fine ailment of remedial Tibet mastiff, in order to offer reference. 1 hair sick condition and clinical expression ill dog are 2~4 month more age young dog, majority is come on suddenly, anorexia or abandon absolutely, mental depressed, systemic symptom worsens quickly, produce acuteness sex vomiting and diarrhoea, puke is first feed rotten, afterwards is yellow or olive bubble mucus and hematic type thing, have diarrhoea dung has the sticky stiff thing of grey yellow, turn after for rare dung of effluvial shape of embedded mucous membrane, then shows hematic dysentery, because ill dog is acuteness vomiting and diarrhoea are rapid dehydration, eyeball cave in, temperature is shown two-way and calorific,℃ of the 40~41 at the beginning of disease, 1~2d falls it is normal to come, after 3~4d answer elevatory, die very quickly next, 4~5d of course of diseases, the temperature when on one's deathbed drops more to normal temperature the following. Bottom of stomach of dog of die in one's bed of 2 analyse check has haemorrhage sex inflammation, show cardinal, small intestine mucous membrane falls off, alvine wall attenuates, there is gules mucus inside, mix inside large intestine have show wine blood excrement and urine, there are a lot of haemorrhage places on mucous membrane, haemorrhage of rectum mucous membrane is more, mesentery lymph node enlargement, sometimes afterwards sends intussusception, alvine tangent plane bleeds, cardiac muscle is loose, color becomes weak, cystic plentiful, liver is qualitative fragile and brittle. 3 diagnose 3.1 epidemiology to diagnose this ill much hair at young dog, rather dog of epidemic disease have an inoculation feels the most easily, be affected directly or affect secondhand, basically pass enteron infection, this disease all can happen all the year round, but with cold...

下面介绍中西医结合治疗藏獒细小病毒性肠炎的心得，以供参考。1发病情况和临床表现病犬多为2~4月龄幼犬，多数为忽然发病，食欲减退或废绝，精神沉郁，全身症状急剧恶化，发生剧烈性呕吐和腹泻，呕吐物先为食糜，继为黄色或黄绿色泡沫黏液和血样物，泻粪有灰黄色的黏稠物，后转为恶臭的内含黏膜状稀粪，继而呈血痢，由于病犬剧烈呕吐和腹泻迅速脱水，眼球下陷，体温呈双向发热，病初40~41℃，1~2d降至正常，3~4d后又复升高，然后很快死亡，病程4~5d，临死时体温多下降至常温以下。2剖检病死犬胃底部有出血性炎症，呈深红色，小肠黏膜脱落，肠壁变薄，内有红色黏液，大肠内混有呈暗红色血液粪便，黏膜上有许多出血点，直肠黏膜出血较多，肠系膜淋巴结肿大，有时继发肠套叠，肠切面出血，心肌松软，颜色变淡，胆囊充盈，肝质脆而易碎。3诊断3.1流行病学诊断本病多发于幼犬，未免疫接种犬最易感，直接感染或间接感染，主要通过消化道感染，该病一年四季均可发生，但以寒。。。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫，第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株；随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛，三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接／酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D、pTARGET／P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1／IFN分别／同时免疫，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：随后将质粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1／IFN同时免疫牛，三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。