p15基因真核表达重组体及人宫颈癌细胞转染的研究

将含人p15cDNA质粒pBS-SK-p15以EcoRI和XhoI进行双酶切，再与真核表达载体pCR^TM3经相同酶切点连接，转化感受态细菌，筛选，鉴定得到可在真核细胞中表达人p15基因的重组质粒PCR-hp15。以脂质体介导法转染人宫颈癌细胞，用G418筛选，得到转染的人宫颈癌细胞，转染细胞的恶变程度有所改善。     

辣椒碱对斜纹夜蛾细胞增殖及凋亡的影响

采用3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四唑(MTT)检测法、荧光显微镜细胞形态观察法和流式细胞仪(flow cytometry,FCM)技术,研究了辣椒碱对离体培养斜纹夜蛾Spodopt-era litura(SL)细胞增殖及凋亡的影响。结果表明:辣椒碱对SL细胞24 h的IC50值为46.89μg/mL,与鱼藤酮差异不显著。40μg/mL辣椒碱处理SL细胞24h后,细胞形态明显不规则,呈现出凋亡的典型症状。10、20和40μg/mL辣椒碱处理SL细胞48h后,细胞早期凋亡率分别为13.56%、18.61%和3.81%,而晚期凋亡和坏死率分别为23.78%、34.28%和61.72%。辣椒碱处理细胞48h后,细胞周期变化明显,细胞生长阻滞于G2/M期。表明辣椒碱对斜纹夜蛾细胞具有良好的抑制作用,能致细胞凋亡并有一定的浓度依赖性。     

云南松松塔提取物对H22实体瘤小鼠模型的抑瘤作用

研究云南松松塔乙醇提取物(PEA)和碱水提取醇沉物(PED)对H22实体瘤小鼠的抑瘤作用。建立H22实体瘤小鼠模型,随机分组,灌胃给药,1次/d,给药10d。末次给药24h后,取血检测γ干扰素(IFN-γ)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)和白细胞介素-10(IL-10)含量;剥离肿瘤组织并称重,计算抑瘤率;检测肿瘤组织匀浆中环氧化酶-2(COX-2)和前列腺素E2(PGE2)含量;石蜡包埋HE染色观察肿瘤细胞病理组织学变化。结果显示,PEA和PED各剂量组小鼠平均肿瘤重量较模型组显著减小;最大抑瘤率分别达46.11%和58.05%。PEA和PED各剂量组小鼠外周血中IFN-γ和IL-2含量较模型组显著升高;IL-4和IL-10含量较模型组显著降低。PEA和PED各剂量组小鼠肿瘤组织中COX-2和PGE2含量较模型组显著减少;给药组正常肿瘤组织减少,有出血和坏死,脂肪组织增多。结果表明,PEA和PED可抑制H22实体瘤小鼠的肿瘤生长,其机制与调节Th1/Th2细胞平衡和抑制COX-2与PGE2的表达有关。     

植物衰老的分子机制及其调控

综述了植物衰老的意义和影响衰老的内部、外部因素 ,并从分子水平上探讨了植物衰老的机制及其遗传学调控     

蜂胶对小鼠T细胞亚群及主要相关细胞因子分泌的影响

为阐明蜂胶对小鼠特异性细胞免疫应答的影响特点,选择1、2、4 mg/ 3个剂量分别经腹腔注射给6周龄Balb/c小鼠,在0、7、10、15 d采血样,采用流式细胞术、ELISA分别测定小鼠外周血CD4+T细胞、CD8+T细胞数量、血清中IFN-γ、IL-4含量.结果显示,与注射蜂胶前相比,各不同剂量的蜂胶均能够使CD4...     

小鼠胚胎干细胞密度梯度离心去核法

在细胞松弛素B的参与下，通过Ficoll密度梯度离心法可有效制备小鼠无核ES细胞，在适宜条件下，悬液培养中40％的小鼠ES细胞可发生去核作用，经Hoechst染色和形态观察确定：80％的去核ES细胞在30min内可恢复其原有形态，但其生活力不会超过48h，这为研究ES细胞的胞质功能及核质互作效应提供了一种可行的细胞去核方法。     

Squamous cell carcinoma in a digit of the hind limb with systemic metastasis in a 17-year-old female koala

We encountered a case of cutaneous squamous cell carcinoma (SCC) in a 17-year-old female koala at a zoo. A fragile, papillary, elevated mass was found on the third digit of the right hind limb. SCC was identified histopathologically: squamous cell-like polygonal tumor cells showed a nest-like growth pattern with epidermal down growth, central keratinization and necrotic foci, and invaded dermal connective tissues. Metastatic lesions were observed in various organs, including the lung and axillary lymph node: in the lung, multiple metastatic foci similar to the primary lesion, and in the axillary lymph node, individual polygonal tumor cells infiltrated the sinusoids. Immunohistochemistry revealed that the tumor cells were positive for proliferating cell nuclear antigen, which exhibited 32–33% of labeling indices in the tumor cells. To our knowledge, this is the first report of a case of SCC in a digit of a koala.     

Growth of Preimplatation Bovine Embryos

Development of mammalian embryos in vitro is functionally and temporally inferior to embryo development as it occurs inside the female reproductive tract. The deficiencies of cultured embryos range from slow cleavage rates to complete developmental arrests or blocks, occurring at particular stages in many species. A variety of approaches have been used to overcome the blocks, including most extensively the coculture of preimplantation embryos with various somatic cells. However, even with coculture, development of embryos in vitro is still not equivalent to that in vivo. In most laboratories, only 25–40% of inseminated oocytes develop into morulae and blastocysts in spite of numerous variations on the basic technique. A better understanding of the factors governing embryonic growth is required before we can hope to achieve results comparable with those occurring in vivo.     

Decreased blood cell counts were not observed in cattle living in the “difficult‐to‐return zone” of the Fukushima nuclear accident

White blood cells, especially lymphocytes, are susceptible to radiation exposure. In the present study, red blood cell, total white blood cell, and lymphocyte counts were repeatedly measured in cattle living on three farms located in the “difficult‐to‐return zone” of the Fukushima nuclear accident, and compared with two control groups from unaffected areas. Blood cell counts differed significantly between the two control groups, although almost all the values fell within the normal range. The blood cell counts of the cattle in the “difficult‐to‐return zone” varied across sampling times even on the same farms, being sometimes higher or lower than either of the two control groups. However, neither a statistically significant decrease in blood cell counts nor an increase in the rate of cattle with extremely low blood cell counts was observed overall. The estimated cumulative exposure dose for the cattle on the most contaminated farm was within a range of 500–1000 mSv, exceeding the threshold for the lymphopenia. Because of the low dose rate on these farms, potential radiation damages would have been repaired and have not accumulated enough to cause deterministic effects.     

果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及风味影响的研究

试验旨在研究果香味剂对奶牛乳腺上皮细胞培养液中乳成分含量及培养液风味的影响。试验选用自制的奶牛乳腺上皮细胞,设5个处理组,果香味剂的添加剂量分别为0、5、10、15和20μg/mL。结果表明:①当培养基中果香味剂添加量为15μg/mL时,乳腺上皮细胞培养液中总蛋白水平、甘油三酯及乳糖含量均达到最高,其中对脂肪含量影响显著(P<0.05),而对蛋白和乳糖含量影响不显著(P>0.05),之后呈下降趋势,说明果香味剂可能会诱导奶牛乳腺上皮细胞乳蛋白、乳脂和乳糖分泌水平的升高;②运用PT/GC-MS分析乳腺培养液中的风味成分,与对照组相比,各试验组中除具有原有乳腺培养液的风味外,还有乙酸异丁酯、乙酸异戊酯、丙酸异戊酯、丙酸戊酯、丁酸异丁酯、丁酸戊酯、丁酸乙酯、邻苯二甲酸二异丁酯等物质成分,而这些物质成分正好是果香味剂中进入到乳腺培养液中的风味物质,且当添加剂量为15μg/mL时,各物质成分含量为最高(P<0.05),之后呈下降趋势。果香味剂能显著影响奶牛乳腺上皮培养液中乳成分含量和风味。