奶牛MTT比色法的最佳条件探讨

为建立奶牛淋巴细胞增殖反应MTT比色法,对试验条件进行了研究。应用L16(45)正交试验,对影响MTT比色法的四个主要因素,包括ConA浓度、细胞浓度、培养液小牛血清浓度及培养时间进行了比较和探索。试验表明几个因素对其增殖反应都有显著影响(P<0.05),且最佳反应条件为:15μg/mL的ConA、1×106/mL细胞浓度、10%小牛血清及60h的培养时间;影响增殖反应的先后顺序为细胞浓度、ConA浓度、培养时间及血清浓度。     

利用体外培养法建立饲料营养成分与瘤胃丙酸产量的回归模型

根据美国康乃尔净碳水化合物和蛋白质体系 (CNCPS) ,分析测定了 4类 2 4种饲料的营养成分 ,利用体外批次培养技术测定了它们在体外培养一定时间后的丙酸产量 ,并由此建立了饲料中产丙酸的营养成分和瘤胃丙酸产量的回归方程     

不同培养条件对鲍鱼菇菌丝生长的影响

通过不同温度、pH值及培养基含水量处理 ,对鲍鱼菇菌丝生长状况进行了研究。结果表明鲍鱼菇菌丝生长适宜的温度范围为 2 0～ 31℃ ,最适温度为 2 5～2 8℃ ;pH值适宜的范围为 4 0～ 7 0 ,最适pH值为 5 5～6 0 ;培养基含水量适宜的范围为 6 0 %～ 80 % ,最适含水量为 80 %。     

Relation between TNF-α,TNFR I expression and apoptosis in oral lichen planus

AIM: To examine the expression and distribution of tumor necrosis factor-α (TNF-α), tumor necrosis factor receptor I (TNFR I) and apoptosis in oral lichen planus, and evaluate their roles and relation in the oral lichen. METHODS: Immunohistochemical technique and TUNEL were employed to study the expression of TNF-α, TNFR I and apoptosis in 50 cases of oral lichen planus and 10 normal oral mucosa specimens. RESULTS: Compared with the normal control group, TNF-α expression was upregulated in mononuclear cells in lamina propria and decreased in keratinocytes in oral lichen planus lesion (P<0.05). On the contrary, TNFR I expression was increased in keratinocytes and decreased in lamina propria in oral lichen planus lesion (P<0.05). The increased apoptosis index in keratinocytes and the decreased apoptosis index in lamina propria were found in oral lichen planus (P<0.05). CONCLUSION: The accelerated apoptosis of keratinocytes and the inhibition of lymphocytes apoptosis may contribute to the formation and progression of oral lichen planus.     

Analysis of blood biochemistry and arterial blood gas of systemic inflammatory response syndrome in the early time after trauma

AIM: To explore the variation of blood biochemistry and arterial blood gas of patients with systemic inflammatory response syndrome (SIRS) in the early time after trauma and improve the diagnosis and first aid. METHODS: Eighty-eight patients with trauma from August 2003 to February 2004 were divided into two groups by their AIS-ISS90 score. The data of temperature, pulse, respiratory rate, white blood cell counts, Hb, blood glucose and arterial blood gas (PaO₂, PaCO₂, HCO₃^-, AG) were collected and compared with each group by statistic methods. RESULTS: Of the 88 patients, 49 underwent SIRS, 12 in light trauma group (ISS≥16) and 37 in severe trauma group (ISS<16). Compared with light trauma group, the data of pulse, respiratory rate, white blood cell counts, blood glucose, AG and rate of SIRS of severe trauma group were higher, PaO₂ and HCO₃^- were lower and the cases of PaCO₂>45 mmHg or <35 mmHg were more (P<0.01). The data of temperature and Hb had not significant difference between two groups (P>0.05). 13 patients had MODS in severe trauma group and 2 died while none had MODS or died in light trauma group. CONCLUSION: Application of AIS-ISS90 and SIRS-related blood biochemistry and arterial blood gas is beneficial for the diagnosis and treatment of patients in the early time after trauma.     

Inhibition of K562 cell growth by antisense drug targeting with VEGF mRNA in vitro

AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs.     

薇甘菊天敌安婀珍蝶生物学特性的研究

安婀珍蝶Actinote anteas(Doubleday ＆ Hewitson)是薇甘菊的天敌,取食薇甘菊Mikania micrantha Kunth.的叶片,能有效的控制薇甘菊的蔓延和生长。在广州市和深圳龙岗进行的观察表明,在实验室条件下,安婀珍蝶一年生长3代~4代。世代平均历期为112.44±1.18天;卵期平均为11.44±1.18天;幼虫期平均为82.54±4.05天;蛹期平均为12.98±1.31天;成虫期平均为7.33±0.80天。本文还记述了该蝶各虫态的形态特征、生活习性及其天敌等。     

引诱剂在综合防治桔小实蝇作用中的评估

以使用引诱剂为主 ,坚持在杨桃园挂引诱笼 2 - 3年后 ,大量降低田间桔小实蝇BactroceradorsalisHendel雄虫数量和卵的受精率 ,配合其它的综防措施 ,防效逐年提高。三个综防点试验实践表明 :杨桃每果虫口数 ,卵孵化率及受害果率大大降低 ,防效达 92 5%。对桔小实蝇的防治初见成效。     

Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37％ at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.     

Effects of inhibition of Kupffer cell and splenectomy on thioacetamide-induced hepatic injury

AIM: To examine the effects of inhibition of Kupffer cell and splenectomy on intestinal endotoxemia and hepatic injury. METHODS: The hepatic injury model was established by treatment with thioacetamide (TAA). At the same time, inhibition of Kupffer cells by intravenous GdCl₃ and splenectomy were performed. Serum alanine aminotransferase (ALT), TNF-α, endotoxin content and phagocytic index were observed. RESULTS: In the TAA+GdCl₃ group, and TAA+splenectomy group, the endotoxin content was significently higher than that in normal and TAA group (P<0.05). The plasma TNF-α, ALT and total bilirubin were significantly lower than those in TAA group (P<0.05). CONCLUSION: Inhibition of Kupffer cells and splenectomy increase plasma endotoxin level, but decreases the plasma TNF-α levels and alleviates the hepatic injury induced by TAA.