Acid–Base Parameters and Steroid Concentrations in Pre‐Ovulatory Follicles and Plasma of Lactating Dairy Cows with Spontaneous and Synchronized Oestrus or Follicular Cyst

The study aimed to compare the acid–base balance and steroid concentrations between follicular fluids (FF) of pre‐ovulatory follicles derived from a spontaneous oestrus (SO), synchronized or induced oestrus (IO) and follicular cysts (CYS) and between FF and blood in dairy cows. Forty‐two dairy cows were included in this study. The animals were allocated to three groups: SO (n = 23); IO (n = 11) using GnRH at day 0 and day 9 and PGF₂α at day 7; and animals with CYS (n = 10). The follicular fluids (FF) were aspirated from the cyst/pre‐ovulatory follicles (? ≥ 15 mm) after SO and after second GnRH dose in IO by transvaginal ultrasound‐guided ovum pick‐up technique. Blood samples (BL) were collected in heparinized vacutainer tubes. The oxygen tension (pO2) in FF of IO was higher (p < 0.05) than in SO and CYS groups. There were negative correlations (p < 0.001, r = ?0.89) between FF and blood pO2. The carbon dioxide tension (pCO2) and lactate level in FF of CYS group were higher (p < 0.05) than in SO and IO groups. There were negative correlations (p < 0.01, r = ?0.73) between blood and FF pO2. Oestradiol‐17β concentration in pre‐ovulatory follicles and plasma of the SO group was higher (p < 0.001) than in IO and CYS groups. Progesterone concentration in pre‐ovulatory follicles and plasma of the SO and IO groups was lower (p < 0.01) than in CYS group. Plasma androstenedione concentration in SO and IO groups was higher (p < 0.05) than in CYS group. In conclusion, acid–base parameters, E2 and P4 levels in the follicular fluid of both IO and CYS groups were deviated greatly from the physiological level (disturbances of intrafollicular/intracystic environment), which may affect the quality of both the oocyte and the granulosa cells.     

Expression of natural killer cell stimulatory receptor NKG2D and its ligand MICA in Tibetan patients with gastric cancer at Qinghai plateau

AIM: To investigate the expression of natural killer cell stimulation receptor D(NKG2D) in peripheral blood and the expression of major histocompatibility complex class I chain-related protein A(MICA) on the gastric carcinoma tissue and the neighboring non-cancerous tissue in Tibetan patients with gastric cancer at Qinghai plateau. METHODS: Samples of 33 patients and 20 healthy persons were collected to detect NKG2D in peripheral blood by Flow cytometry analysis. The expressions of MICA in part of the correspondent tissues were examined by RT-PCR and immunohistochemistry.RESULTS: The expression of NKG2D in the plateau of Tibetan patients with gastric cancer group (13.47 ±5.26)% was significantly lower than that in healthy groups (32.62±10.08)%. The mRNA level of MICA in gastric cancer was significantly higher than that in neighboring non-cancerous tissue (P<0.05). The expressions of MICA proteins showed a significant difference between the neighboring non-cancerous tissue (21.21%, 7/33) and the gastric carcinoma tissue (78.79%, 26/33), between the high differentiation (60.00%, 3/5) and the moderate (72.73%, 8/11), low differentiation (88.24%, 15/17). The expressions of MICA were not correlated with tumor size, sex, age, lymph node metastasis of the patients (P>0.05). Spearman rank correlation displayed that the expressions of MICA mRNA were positively correlated with the MICA protein level (r=0.903, P<0.01).CONCLUSION: The immune escape in Tibetan patients with gastric cancer at Qinghai plateau probably relates to the down-regulation of NKG2D and the up-regulation of its ligand MICA. The activity of NK cells and the anti-cancer cellular immunity level are descending in the plateau of Tibetan patients with gastric cancer. The decrease in the receptor NKG2D is a reason for the activity of NK cell descending.     

番茄木虱取食对含Mi-1.2番茄抗氧化酶活性及防御反应相关基因表达的影响

Mi-1.2基因编码CC-NBS-LRR家族抗病蛋白,对线虫及刺吸式昆虫具有广谱抗性。以番茄木虱和携带Mi-1.2基因的番茄品系‘Motelle’作为研究体系,研究番茄木虱取食对番茄叶片抗氧化酶活性及防御反应相关基因表达的影响。结果表明：番茄木虱的取食显著提高了番茄叶片抗氧化酶POD、CAT、APX等的活性,同时诱导Mi-1.2基因表达;番茄木虱的取食上调SA合成有关基因PAL、SA信号途径标志基因PR-1（P4）及与Mi-1.2功能相关基因WRKY70、WRKY72a、WRKY72b的表达。JA/systemin信号途径和伤害有关的PinII、TPI-1、PR-6、PR-7、PR-5等及Prosystemin编码基因Psy的表达未发生变化;JA合成途径相关酶编码基因AOS2、LoxD下调表达,AOC的表达没有变化。SA信号途径可能参与Mi-1.2介导的番茄对于刺吸式昆虫番茄木虱的防御反应。另外,Mi-1.2的RNAi沉默不会改变上述基因的mRNA水平,说明Mi-1.2基因产物对这些基因的表达无调控作用。     

不同氮肥施用量对青花菜氮肥利用率、氮素含量及产量的影响

为了探明不同氮肥施用量对青花菜氮肥利用率、氮素含量和产量的影响,试验设置5个水平,分别为667 m2施0、5、10、20、30 kg,利用相关性分析及回归方程了解氮肥对青花菜的影响。结果表明,氮肥施用量与氮肥利用率间呈显著负相关,与花球中氮素含量呈极显著正相关。青花菜的氮肥效应方程为:Y=-0.829 5X2+33.17 1X+543.55,在其他条件相同的情况下,667 m2氮肥施用量为19.46 kg,青花菜达到最大收益。     

大肠杆菌不耐热肠毒素B亚单位在大肠杆菌中的表达及其特性研究

将大肠杆菌不耐热性肠毒素B亚单位(LTB)基因eltb克隆到表达载体pET-28a中,转化大肠杆菌BL21/DE3/plyss。采用SDS-PAGE和Western blot分析,结果表明,在大肠杆菌中高效表达了LTB蛋白,其产量约为细菌总蛋白的20%,GM1-ELISA方法鉴定目的重组蛋白可与GM1结合,具有潜在的佐剂活性;由纯化的重组蛋白免疫小鼠制备的抗血清效价为1:16000,并且可以特异性识别天然毒素。结果表明,所表达的LTB有良好的抗原性和佐剂活性,制备的抗血清具有良好的免疫活性,为下一步以LTB作为佐剂的黏膜疫苗研究提供基础材料。     

α-蒎烯合成2-羟基-3-蒎酮的研究

研究了高锰酸钾氧化α-蒎烯合成2-羟基-3-蒎酮的工艺条件,对助溶剂等影响因素进行了探讨,较优工艺条件为:以叔丁醇为助溶剂,不改变pH值,温度0-5℃,KMnO4与α-蒎烯的物质的量比为1.7:1～1.8:1,先加α-蒎烯后加KMnO4.所得2-羟基-3-蒎酮采用IR、MS、1H NMR、3C NMR进行了表征,产率达60%,与文献相同,助溶剂改用叔丁醇后回收率＞90%,高于原用助溶剂丙酮(回收率70%).     

淡水养殖水化学实验课教学改革

为了培养学生的实验技能和科研意识,提高综合能力,笔者对淡水养殖化学实验课教学改革展开探讨。目前淡水养殖化学实验课主要存在实验项目部分重复,实验试剂涉及一些贵重的、有毒的药品等问题。因此,在考虑到不重复设置实验项目及尽量避免有毒试剂大量使用的前提下,笔者结合农业分析测试中心的大型精密仪器设备,尝试实验课的教学改革,注重引进大型精密仪器,整合实验项目,向学生灌输环保安全的理念,培养学生的科研素质,提高学生的综合能力。     

用β-蒎烯为原料合成高纯度对异丙基苯酚

研究了以松节油中β-蒎烯为原料,通过氧化、酸催化开四元环和脱氢三步反应,合成了高纯度对异丙基苯酚,总产率达到31％。并对酸催化开四元环和脱氢步骤中各因素对反应的影响进行了探讨。发现诺蒎酮开四元环转化反应选择在40％的磷酸溶液催化下,80℃反应1h效果较好,产率达到67．9％。4-异丙基环己烯酮与硫脱氢生成对异丙基苯酚,反应选择密封反应2h效果较好,产率达到63％。