接种后的

Allantois and surrounding tissues of 8.5 day post coitum embryos, the hindhut and surrounding tissues of 9.5 dpc and 10.5 dpc embryos, the gonadal ridges and surrounding tissues of 11.5 dpc and 12.5 dpc embryos were collected and digested with 0.25%pancreatin-0.02%EDTA, then the cells were cultured in the plastic petridishes which are pretreated with 0.1% gelatine.

实验方法：取胚龄8.5 d胎鼠尾端尿囊及周围组织，取胚龄9.5~10.5 d胎鼠后肠及周围组织，取胚龄11.5 d，12.5 d胎鼠生殖嵴及周围组织，分别经0.25%胰酶-0.02%EDTA消化并接种于0.1%明胶包被的培养皿中。

The stem was cut into several segments away from the pine seeding, arranged and collected in the order of chronology time and distance from where they were inoculated. To separate B.xylophilus coarsely out from the stems and made them into paraffin slices. And be observed under the potics microscope by dyed with safranin-fast green contrast-stain method and PI fluorescence stain method. In order to find out how about the nematode rigour distributing , but also ebb and flow of their amounts after invading the host pine seeding. Seek the pathological changing orderliness of host cells in different position along with the remotion of nematodes. By deeply analysing the variety of host cell nucleolus to estimate whether the host cell took place the programmed cell death after inoculated B.xylophilus.

按时间进程和与接种点上下距离的远近，对松苗分茎段进行粗分离和制作成石蜡切片，分别用番红-固绿双重染色法和碘化丙锭荧光染色法在光学显微镜下观察，以了解线虫侵染后在寄主体内的准确分布和数量的消长，寻找随着线虫的移动扩散，寄主不同部位细胞病变的数量、速度发生的变化规律，并进一步通过分析其细胞核的变化来判定松材线虫侵染后是否能诱发寄主细胞发生程序性死亡(programmed cell death，PCD)。

Depositing cells were incubated in high-sugar DMEM culture medium containing 15% fetal bovine serum, 100 U/mL penicillin, and 100 U/mL Streptomycin, then cells were seeded in a 25 T culture bottle after cell density was adjusted to 1×109 L-1, and incubated at 37 ℃, 0.05 volume fraction CO2 with saturated humidity. After 72 hours, the culture medium was replaced, and red blood cells and other nonadherent cells were removed. Cells began to passage at 70%-80% confluence.

实验方法：穿刺抽取15 mL脐带血，密度梯度法分离吸取分层界面的白色环形絮状物，其中富含单个核细胞，离心后沉积细胞用含15％胎牛血清、100 U/mL青霉素、100 U/mL链霉素的DMEM高糖培养基悬浮，调整细胞密度为1×109 L-1接种于25 T培养瓶内，放置在37 ℃、体积分数为0.05的CO2饱和湿度培养箱中培养，72 h后更换培养基，去除红细胞及其他未贴壁细胞，至70％~80％融合时传代。

The comparison test of this new methodwith spray inoculation method was conducted. The results showed the former method could increase the incidence ofdisease and uniformity than the latter one. Used cotyledon-brushing with conidial suspension and fungicide foliarspraying method, the EC 50 values of hexaconazole, myclobutanil, triadimefon, thiophanate-methyl and chlorotha lonil to Sphaerotheca fuliginea treated in different time after inoculation were determined; the results showed that 96 h was better time for disease severity evaluating.

比较了白粉病菌分生孢子悬浮液涂抹法和喷雾法接种黄瓜幼苗子叶的效果，结果表明，涂抹法发病率高，均匀度更好；测定了接菌后不同时间施药，白粉病菌对己唑醇、腈菌唑、三唑酮、甲基硫菌灵和百菌清等 5 种杀菌剂的敏感性，结果表明，接菌后 96h 施药较为敏感，测得的 EC 50 较小。

METHODS: Bone marrow was sterilely separated from human. After heparinization, human BMSCs were harvested using density gradient centrifugation and adherence method. At the fifth passage, BMSCs at 1×108/L were incubated in the 6-well plate and divided into 2 groups. BMSCs in the edaravone group were 50% confluent and incubated in L-DMEM containing basic fibroblast growth factor and fetal bovine serum for 24 hours. After washing in PBS, these BMSCs were incubated in serum-free L-DMEM containing 20 mg/L edaravone for 24 hours. BMSCs in the blank control group were incubated in L-DMEM, supplemented with 10% fetal bovine serum.

无菌抽取的骨髓经肝素化后，采用密度梯度离心法及贴壁筛选法分离获得人骨髓间充质干细胞，传至第5代按1× 108 L-1接种于6孔板内，设立2组，依达拉奉组细胞达50%融合时用含碱性成纤维生长因子、胎牛血清的L-DMEM预诱导24 h，PBS洗涤后再用20 mg/L依达拉奉无血清L-DMEM诱导24 h；空白对照组始终用含体积分数为10%胎牛血清的L-DMEM培养，不加任何预诱导剂和诱导剂。

Sp. tritici conidiospores. 40 h after inoculation, penetration of the fungus and formation of haustoria in transformed cells were observed to evaluate the effect of the target gene's product on the invasion of the fungus.

基因枪轰击后高密度接种白粉病菌孢子。40h后观察转化阳性表皮细胞度其表面抱子的发育，并以阳性转化细胞中白粉病菌成功侵入的细胞所占比例为指标分析目标基因的表达对白粉病菌入侵及吸器形成产生的影响(对照为单独导入GUS基因的表皮细胞)。

METHODS: Normal human BMSCs were isolated and cultured by the whole bone marrow method. Cells of the third passage at 1×108/L were incubated on coverslip in a 6-well plate, and randomly divided into 3 groups: osteogenic induction group, which was added with primary medium and osteogenic inducer, supplemented with 10-8 mol/L dexamethasone, 10 mmol/L β-phosphoglycerol and 50 mg/L vitamin C; leptin osteogenic induction group was added with 50 nmol/l leptin in addition to osteogenic inducer. Cells in the blank control group were only treated with LG-DMEM containing 10% fetal bovine serum.

采用全骨髓法体外分离培养人骨髓间充质干细胞，传至第3代以1×108 L-1密度接种至预置盖玻片的6孔培养板中，设立3组：成骨诱导组添加原代培养液后，再加入含10-8 mol/L地塞米松、10 mmol/L β-磷酸甘油、50 mg/L维生素C的成骨诱导培养基；成骨+瘦素诱导组添加成骨诱导培养基后，再加入50 nmol/L瘦素；空白对照组仅加入含体积分数为10%胎牛血清的LG-DMEM培养液。

ES cells were maintained on gelatin-coated dishes in Iscove's modified Dulbecco medium containing 0.1 mmol/L of nonessential amino acids, 2 mmol/L of L-glutamine, 0.1 mmol/L of monothioglycerol, 15% fetal bovine serum, and 1000 U/mL leukemia inhibitory factor. Embryonic bodies were firstly formed by ES cells with hang drip method in IMDM without LIF. Six-day-old EBs were trypsinized and the cells were resuspended at a concentration of （3-4）×109 L-1 cells per 1 mL of a 1:1 mixture of culture medium and Matrigel?

实验方法：将鼠胚胎干细胞株接种于被覆明胶的培养瓶内，加入含0.1 mmol/L非必需氨基酸、2 mmol/L L-谷酰胺、0.1 mmol/L二羟基丙硫醇、15%胎牛血清、1 000 U/mL白血病抑制因子的IMDM培养液。3 d后采用&悬滴法&使细胞在不含白血病抑制因子的上述IMDM培养液中形成胚胎体，6 d后胰酶消化得到胚胎干细胞，浓度调整为（3~4）×109 L-1，与Matrigel？

Methods The retinae of 16 postnatal 2～3 day Sprague-Dawley rats were dissected into cell suspension with trypsin digestion.

采用胰酶消化法将16只生后2～3天的Sprague-Dawley 大鼠视网膜制成细胞悬液后，接种于涂以鼠尾胶原的24孔培养板，预先置入1 cm×1 cm的载玻片。

Pneumophila serogroup 5 antibody to develop two immuo-colloidal gold tests respectively. Main works listed below as:1、Preparation and identification of polysaccharide antigen of L. pneumophila Bacteria of LP1 ~ 7,9 and 10 were cultured on yeast agar buffer activated carbon for three to five days at 37℃, 5%CO2. Protein-free polysaccharide antigens were obtained after harvest in cells, extraction, deproteinization, dialysis and other steps. Their immunogenicities were verified by ultraviolet spectrophotometer full-wavelength scanning and Western blotting.2、Preparation and identification of rabbit anti-LP1 antibodies and rabbit anti-LP5 antibody Rabbit anti-LP1 and anti-LP5 antibodies were purified after rabbits were immuned with antigens isolated as described above. The purities of both antibodies were above 80% and the titer of blood serum 1:32 tested by double antibody sandwich assay.3、Development of colloidal gold immunochromatographic assay kit The size of colloidal gold particles in the kit was 25nm. The optimal concentrations for antibodies were 30μg/ml and the sensitized concentrations of NC membrane were 5 mg/ml.

主要研究工作从以下几个方面进行：1、LP1~7、9和10型多糖抗原的制备与鉴定将LP1~7、9和10型菌株分别接种在缓冲活性炭酵母琼脂培养基上，37℃、5%CO2的条件下培养，3~5天后洗下菌苔，经抽提、除蛋白、透析等步骤后得到基本无蛋白的LP多糖抗原，经紫外分光光度仪全波长扫描及Western blotting验证其抗原良好。2、兔抗LP1抗体和兔抗LP5抗体的制备与鉴定分别用LP1、LP5型多糖抗原免疫家兔，采用琼脂糖双向扩散试验检测，两种抗体血清效价均为1：32；饱和硫酸铵法提取抗体，SDS-PAGE检定其抗体纯度均达到80%以上。3、胶体金免疫层析检测试剂的初步研制采用柠檬酸三钠还原法制备约25nm大小的胶体金颗粒；分别制备兔抗LP1抗体、兔抗LP5抗体的金标探针，两种抗体的最适标记量均为30μg/ml；选择适当孔径的微孔滤膜为载体包被两种抗体，NC膜包被浓度均为5mg/ml。

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。