人肺精确薄片体外移植培养模型的建立及对流感病毒感染的反应

目的建立人肺精确薄片体外移植培养模型;并检测暴露于流感病毒下的人肺薄片里是否存在感染和病毒复制。方法参照改良琼脂糖充填肺组织的方法,以1.5%低熔低胶琼脂糖经充膨、钻取、切片及培养等步骤建立人肺精确薄片体外移植培养模型。选取不同时间点对移植培养的人肺薄片进行显微镜下组织形态观察;以乳酸脱氢酶释放毒性实验评价细胞毒性;以RT-PCR和免疫组织化学方法测定人肺薄片组织对暴露的流感病毒的反应。结果人肺精确薄片模型经长时间(>10d)的培养,维持正常的结构和功能,支持人肺薄片组织暴露于流感病毒后的感染和病毒复制。结论建立的人肺精确薄片体外移植模型可靠和稳定,可被用于流感病毒与人体肺器官感染的研究。     

Induction of Chemokine and Cytokine Responses by Helicobacter pylori in Human Stomach Explants

Background: The cytokine response during the acute phase of Helicobacter pylori infection in humans has not been studied. The aim of this study was therefore to investigate the early cytokine responses against H. pylori using cultured human stomach explants as a model of acute infection. Methods: Gastric corpus tissue obtained from 13 adult uninfected and 3 H. pylori-infected patients undergoing gastric surgery due to obesity was used for preparation of mucosal explants. The cultured explants were exposed to different H. pylori     

速尿对离体培养豚鼠血管纹毒性作用的观察

目的：观察速尿对离体培养的豚鼠血管纹组织的影响，探讨速尿耳毒性的作用机制。方法：20只花色豚鼠随机分成二组：速尿组（n=16），正常对照组（n=4）。应用组织块培养的方法，将血管纹组织培养24小时，随即对不同的实验组分别应用不同终浓度的的速尿（60、300、600、1250、2500μg/ml），分别继续培养30分钟和90分钟，观察培养的血管纹组织学结构。结果：速尿组在组织学方面均未出现血管纹水肿、缘细胞胞浆肿胀、细胞间隙扩大和中间细胞皱缩等病理改变，与对照组相比较血管纹结构无显著性差异。结论：速尿对体外培养的豚鼠血管纹组织无明显诱导水肿的作用，提示速尿耳毒性的产生，可能是间接的作用机制。     

Stimulated mouse ear epidermis in explant culture —The effect of retinoic acid and hexadecane

Summary Ear skin of adult mice was used as a source of explant cultures of epidermal cells and the requirements for growth were defined. Two growth phases were observed, the first more rapid than the second. All-trans-retinoic acid or n-hexadecane was applied topically for up to 9 days and the skin was then used for explant cultures. During both treatments the ear became red and swollen. The epidermis became hyperkeratotic or deprived of its horny layer. The thickness of the stratum Malpighii increased to up to nine layers, from the normal of two or three layers. The stratum granulosum became multilayered and keratohyalin granules were abundant. Explants of n-hexadecane-treated skin behaved exactly as their normal controls. All-trans-retinoic acid induced an increase in migratory cell activity during the first growth phase. In the second growth phase the growth rate was similar or lower than that of the controls.Supported by the Karolinska Institutet, the Finsen and E. O. and Edla Johansson Foundations, the Swedish Psoriasis Association and the Swedish Medical Research Council (12x-5665)     

植块联合培养法探讨备用背根猫脊髓背核组织对鸡胚背根节神经突起生长的影响

５只单侧备用背根猫，于术后第５ｄ分别切取手术侧、非手术侧脊髓Ｌ_３节段背核组织植块，与Ｈａｍｂｕｒｇｅｒ３５期鸡胚背根节（ＤＲＧ）进行植块联合悬滴培养，以单独ＤＲＧ培养作为参照。于培养２４ｈ、４８ｈ观察ＤＲＧ神经突起的生长情况并测量其长度。结果发现：１．所有ＤＲＧ从培养２４ｈ到４８ｈ其神经突起都有明显增长；２：同一观察时间内，参照组与对照组（ＤＲＧ与非手术侧背核植块联合培养组）神经突起的生长状况较为相似，神经突起少而短，从ＤＲＧ迁移出的细胞多。而实验组（ＤＲＧ与手术侧背核植块联合培养组）神经突起多且长，从ＤＲＧ迁移出的细胞少；３：在分别求出每批培养物的实验组、对照组ＤＲＧ神经突起平均长度与参照组者的比值及５批培养物的平均比值后，发现两个观测时间实验组的平均比值都明显大于对照组者。本研究结果提示，猫脊髓在部分腰骶背根切除后，背核组织的促神经突起生长作用增强。     

Cytokinin induced shoot regeneration and flowering of Scoparia dulcis L. (Scrophulariaceae)-an ethnomedicinal herb

Objective

To develop an improved protocol for micropropagation of ethnomedicinally important Scoparia dulcis (S. dulcis) L.

Methods

Explants were inoculated on MS basal medium supplemented with kinetin and 6-benzylaminopurine for shoot bud induction. To enhance the shoot induction, various auxins like 3-indoleacetic acid or 3-indolebutyric acid or α-naphthylacetic acid were tested along with 2.32 M KI and 4.44 µM BAP. The regenerated shoots were rooted in half strength MS medium supplemented with various concentrations of IAA, IBA or NAA. After roots were developed, the plantlets were transplanted to pots filled with vermiculate and sand and kept in growth chamber with 70%–80% humidity under 16 h photoperiod. After acclimatization, the plantlets were transferred to the garden and survival percentage was calculated. Data were statistically analyzed and means were compared using Duncan''s multiple range test (P<0.05).

Results

An in vitro method was developed to induce high frequency shoots regeneration from stem, mature leaf and young leaf explants of S. dulcis. Shoot induction on young leaf explants was most successful in MS medium supplemented with combination of two cytokinins (2.32 µM KI and 4.44 µM BAP) 2.85 µM IAA, 10% CM and 1 483.79 µM adenine sulfate. A single young leaf explant was capable of producing 59 shoots after 13 days of culture. Flower was induced in medium supplemented with combination of KI and BAP.

Conclusions

Cytokinins are the key factor to induce the direct shoot regeneration and flowering of S. dulcis.     

组织贴块法人的支气管上皮细胞的原代培养

目的 通过组织贴块法建立人的支气管上皮细胞原代培养的方法.方法 采用无血清支气管上皮培养基,对经手术切除获得的支气管进行组织贴块法培养获得的细胞进行原代培养和传代,通过倒置显微镜以及细胞免疫化学观察和鉴定细胞.结果 此方法获得的细胞成活率高、纯度高,细胞呈扁平、多边形,象铺路的鹅卵石样分布.细胞角蛋白表达阳性.结论 组织贴块法是一种简便、有效的培养人的支气管上皮细胞的方法,无血清培养基可提供满意的生长条件,提高上皮细胞的纯度,为进一步研究不同的呼吸系统疾病提供了良好的模型.     

用于神经突导向研究的螺旋神经元培养体系初步观察

目的 探寻适用于螺旋神经元神经突导向研究的螺旋神经元培养体系.方法 取新生SD大鼠的螺旋神经节,分别进行分离培养和组织块培养,比较神经突的生长模式.组织块培养时,采用不同底物(包括多聚赖氨酸、层粘连蛋白和I型胶原蛋白等)铺片,观察对螺旋神经元组织块贴壁和神经突生长的影响.用微管相关蛋白2(MAP2)抗体和微管聚合蛋白1(Tau-1)抗体鉴别神经突的极性.结果 组织块培养体系中,部分神经突长出细胞毯,在无干扰的环境中生长.以多聚赖氨酸加层粘连蛋白为底物铺片时,组织块贴壁率高,神经突数量多、生长较快,有利于获取更多适于神经突导向研究的神经突.培养体系中的神经突可被Tau-1抗体标记,而不能被MAP2抗体标记.结论 在多聚赖氨酸加层粘连蛋白包被的可移动玻片上,进行螺旋神经元组织块培养获取自由神经突的方法 适合螺旋神经元神经突的导向研究.     

Quality of placental RNA: Effects of explant size and culture duration

We evaluated the impact of placental micro (≤50 mg) and macro (∼200 mg) explants, oxygen concentration and culture method on placental RNA quality after long-term culture. Our findings show that micro explants cultured at 8% oxygen have the best RNA quality and tissue structure. Macro explants were less viable after long-term culture. Macro explants and explants undergoing syncytial degeneration produced poor quality RNA and should be avoided.     

Vasoactive intestinal peptide and gastrin-releasing peptide play distinct roles in the suprachiasmatic nucleus

Vasoactive intestinal peptide and gastrin-releasing peptide levels were measured in the punched-out suprachiasmatic nucleus tissue from rats kept under a prolonged dim light (in vivo). Vasoactive intestinal peptide content increased from 4 to 8 h, returned to the baseline level at 12 to 16 h, and then increased again until 36 h after the light was switched off (dim light). Gastrin-releasing peptide level, in contrast, showed no significant changes, but a slight decrease from 1 to 4 h was detected under the dim light. In suprachiasmatic nucleus explant-slice culture, in vitro, Arg-vasopressin release increased transiently or showed a decrease at 30 min after exposure to vasoactive intestinal peptide or gastrin-releasing peptide, respectively. Treatment with anti-vasoactive intestinal peptide and anti-gastrin-releasing peptide antibodies reversed these effects. These findings suggest reciprocal roles of vasoactive intestinal peptide and gastrin-releasing peptide in Arg-vasopressin release.