染色法

Methods Seventy-four strains of Salmonella enterica serovar enteritidis (S. enteritidis), S. pullorum and S, gallinarum isolated from chickens were determined for biofilm using crystal violet staining, and the strain C050041 with well biofilm formation was chosen to construct a mutant library using transposon mutagenesis.

利用结晶紫染色定量法对74株鸡源的肠炎、鸡白痢和鸡伤寒沙门氏菌进行生物膜测定，选择生物膜生长较好的肠炎沙门氏菌C050041，采用转座子随机插入法构建突变株库。

Results:①The amount of human colon carcinoma cell line SW480 treated by quercetin decreased. The morphology of partial SW480 cells was shrunk volume, integrated cell membrane, condensed cytoplasm, pyknotic chromatin, nuclear fragmentation. Apoptotic Corpuscles were found by electron microscope.②MTT colorimetric assay showed quercetin inhibited the growth of human colon carcinoma cell line SW480 in a time- and dose-dependent manner when the concentration of quercetin was 30、60、90μmol/L.③Flow cytometry analysis showed the cell cycle of SW480 cell was restricted in G1/S. G0/G1 phase rate increased and S phase rate decreased with increasing concentration of quercetin and time lasting.④ Zymogram analysis assay showed the secretion of matrix metalloproteinases in human colon carcinoma cell line SW480 treated by quercetin decreased. With increasing concentration of quercetin, the secretion of MMP-2 and MMP-9 decreased.⑤Immunohistochemistry method demonstrated the position expression of Cathepsin-D in SW480 cell was suppressed by quercetin in a time- and dose-dependent manner.

研究结果：经槲皮素处理的人结肠癌SW480细胞数量减少，部分细胞体积缩小，细胞膜完整，胞浆浓缩，核染色质固缩，细胞核碎裂，形成凋亡小体；MTT法检测显示当作用浓度为30μmol/L~90μmol/L时，槲皮素对人结肠癌SW480细胞的生长有抑制作用，其抑制作用随着作用浓度的增加和作用时间的延长而增强；流式细胞学发现槲皮素主要作用于人结肠癌SW480细胞周期的G1/S期，大部分细胞被阻断于S期，随药物浓度的升高和作用时间的延长，G0/G1期细胞比例逐渐增加，S期细胞比例逐渐减少；酶谱分析法检测显示不同浓度的槲皮素能够抑制人结肠癌SW480细胞分泌MMP-2及MMP-9，随浓度的升高，MMP-2及MMP-9的分泌量减少；免疫组织化学法显示不同浓度的槲皮素处理人结肠癌SW480细胞后，Cathepsin-D的表达随药物浓度的升高和作用时间的延长而降低。

Methods 24 SD rats were randomly divided into normal group,model group,aminophylline treatment group(35 mg/kg), 8 rats in each group. The bronchial asthma model in rats was built by egg protein sensibilization and longterm inhalation provocation. The rats in treatment group were lavaged each day from the first time of provocation.After 4week treatment, the rats were killed to obtain lung tissue for staining with HE.

24只SD大鼠随机分为正常组、模型组、治疗组，每组8只，除正常组外以卵蛋白致敏并吸入激发法制备大鼠哮喘模型，治疗组、模型组从第1次哮喘激发开始(造模第3周)分别给予氨茶碱、0.9%氯化钠溶液灌胃给药，1次/d，用药4周处死大鼠，取肺组织HE染色，用彩色图像分析仪测量支气管壁面积、支气管平滑肌面积，用ELISA双抗体夹心法测定肺组织MMP9和TIMP1含量。

The bronchial asthmatic model was established by egg protein sensibilization and longterm inhalation provocation.The rats of each treatment groups were lavaged each day from the first time of provocation to execution.After 4 weeks of treatment,the rats were killed to obtain lung tissue for dyeing of HE.A computer assisted image analysis system was used to determine the thickness change of bronchus wall and smooth muscle.The contents of MMP9 and TIMP1 in lung tissue were determined by ELISA double antibody sandwich method.

除正常组外以卵蛋白致敏并吸入激发法制备大鼠支气管哮喘模型，各治疗组均从第1次哮喘激发开始(造模第3周)至处死前每天灌胃给药，激发并给药4 w后处死大鼠，取肺组织HE染色，彩色图像分析仪测量支气管管壁厚度、平滑肌厚度，采用ELISA双抗体夹心法测定肺组织MMP9、TIMP1含量。

Six rats of each group were randomly sacrificed on day 7,14,28 and 42 respectively.The histological changes of lung tissue were studied by HE and Masson's trichrome staining.Hydroxyproline level in lung tissue was measured by digestion method.Protein and mRNA expression of transforming growth factor-β1（TGF-β1） were assayed by immunohistochemistry and RT-PCR respectively.

各组分别于造模后第7、14、28及42 d随机处死6只，取肺组织行HE、Masson染色评价肺组织病理变化，消化法测定肺组织羟脯氨酸含量，分别采用RT-PCR和免疫组化法测定肺组织转化生长因子β1（TGF-β1）的mRNA和蛋白表达。

The thymuses, spleens, sera and anticoagulant blood were collected after the rats were killed, to carry out these works:(1) Zinc concentrations were analyzed by atomic absorption spectrophotometry;(2) Body weight and the weight of thymuses and spleens were measured, the organ index were calculated;(3) Changes in pathology of thymus and spleen were observed;(4) Using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) method, the apoptosis of thmocytes and spleen lymphocytes was checked;(5) The expression of bcl-2、bax mRNA in thymus and spleen were detected by RT-PCR (reverse transcription polymerase chain reaction);(6) The expression of p56〓, a signal transduction protein in thymus and spleen were detected by immunohistochemistry;(7) Two-color cytofluorometric analysis was used to assess the expression of CD4, CD8, CD45RA and CD45RC in peripheral blood lymphocytes.

动物处死后留组织、抗凝血及血清，进行以下检测：(1)原子吸收法测血清锌浓度；(2)测胸腺、脾脏脏器重量并与大鼠体重比较，计算脏器指数；(3)HE染色对胸腺、脾脏进行病理观察；(4)原位末端标记法测胸腺、脾脏中淋巴细胞的自发凋亡，并通过图像分析进行比较；(5)逆转录聚合酶链式反应检测凋亡调控基因bcl-2、bax mRNA在免疫器官中的表达；(6)免疫组化方法检测信号转导蛋白p56〓在免疫器官中的表达；(7)流式细胞术分析外周血淋巴细胞中CD4+、CD8+、CD45RA+及CD45RC+CD4+、CD45RC-CD4+细胞的数量和比例。

The rats were killed by cutting theirnecks and physical blood was taken for preparing serum, interleukin 2 (IL-2) was detected by radioimmunoassay ; the spleen was taken for measuring ConA-induced reaction of lympho- cyte proliferation and induced production of IL-2; and the activity of induced production of spleen's IL-2 was determined by the means of cell proliferation of mouse thymus; the skin and trachea were taten for slice with paraffin wax, dyeing with methybenzoamine blue so as to survey mast cells .

将大鼠断头留取躯干血制备血清，用RIA法检测白细胞介素2（IL-2）；取脾脏行ConA诱导的淋巴细胞增殖反应测定和IL-2诱生；并用小鼠胸腺细胞增殖法检测脾IL-2诱生活性；取皮肤和气管做石蜡切片、甲苯胺兰染色检测肥大细胞，结果显示：应激组较对照组脾淋巴细胞增殖降低，cpm分别为9466±3.17和24019±3879（P.001）；MC明显减少，分别为8.6±3.0/HP和25.4±3.7/HP（P.001），前组形态异常较其甚（MC多较小，呈多角或不规则状），且组织中可见分散的崩解颗粒。

This study was conducted to examiune the fibrotic effect of Ni-Ti and 317L al loys in esophagus.The extract fluid from Ni-Ti,317L alloys was made according t o the ASTM standards of U.S.A. The Fb of esophageal scar was cultured primarily ,then incubated with alloy abstract fluid. The proliferating activity of Fb was measured by MTT at 4, 24, 48, 72 hours in the course of culturing. The esophagu s embedding test of Ni-Ti,317L alloys was made according to ASTM standards of U .S.A.The tissue around the alloys was taken at weeks 2 and 12,and the pathologi c changes were analysed.

为探讨新型支架材料Ni-Ti、317L合金在食管局部的致纤维化作用，按美国ASTM标准制备NiTi、317L合金的金属浸提液；&组织块培养法&原代培养食管壁疤痕的成纤维细胞，传代后以金属浸提液进行培养，分组后分别培养4、24、48、72 h，MTT法检测不同培养时间后Fb增殖功能的变化；按美国ASTM标准进行NiTi、317L合金试件的食管壁内包埋实验，即将金属试件经表面处理后直接置入食管壁粘膜层与肌层之间，术后2、12周取出包埋组织，分析试件周围组织的病理变化，并进行胶原纤维染色，观察纤维形成状况。

The proliferation of cells were detected by trypan blue exclusion, methyl thiazolyl tetrazolium assay, Hoechst33342 fluorescence staining and flow cytometry method.

利用台盼兰排斥试验、噻唑蓝比色法、Hoechst33342 荧光染色、流式细胞术测细胞周期法等实验技术分别检测了不同浓度AA 和DHA 对大鼠前体脂肪细胞活力、细胞周期及凋亡的影响。

objective the aim of this study is to investigate the expression and the distribution of the nerve growth factor during the period of neural tube development of human embryo.method early development of neural tube was studied in human embryos about 35 gestational days by using immunocytochemical abc technique.result there were ngf immuno-positive substances in the cytoplasm and nuclei of neuroepithelial cells in the ventricular zones of neural tube.in the intermediate zone of neural tube,ngf immunoreactivity was detected in the nuclei of some neurons,or the processes of other neurons which contained no ngf-immunoreactive substances in their nuclei;the expression pattern of ngf in the marginal zone of neural tube was similar to that of the intermediate zone.the density of ngf-immunorecative particles was higher on the rostrum side of neural tube than on the caudal side.the ngf immuno-positive cells were also observed among the somites of embryo under the neural tube.conclusion these results suggest that ngf was an important signal molecule to induce neural tube differentiation,and that ngf may play a significant role in regulation of the biological function of neurons in developing neural tube.

目的 研究捷安肽素的抗真菌作用机理。方法采用形态学方法和同位素标记法。显微形态观察经捷安肽素处理后的供试真菌的形态学变化。进一步采用14c同位素标记的特异底物&尿苷二磷酸-（14c）-葡萄糖&示踪，研究捷安肽素对真菌(1，3)-β-d-葡聚糖合成酶活性反应的影响。结果研究神经生长因子在早期人胚神经管发育过程中的定位表达。方法采用免疫细胞化学 abc法染色，研究35天人胚的发育情况。结果在人胚神经管的室管带中，神经元的细胞质和细胞核ngf免疫反应阳性；在中间带，一部分神经元的细胞核ngf免疫反应阳性，另外一部分神经元的细胞核ngf免疫反应阴性，而其突起ngf免疫反应阳性；在边缘带ngf的表达与中间带相似。在神经管的头侧ngf阳性反应较强，神经管的尾侧ngf阳性反应较弱。结论 ngf在人胚神经管免疫反应阳性，表明ngf可能是诱导神经管分化发育的重要信号分子，提示ngf可能在人胚神经管的发育中具有十分重要的作用。神经生长因子；人胚；神经管；发育

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。