检测心肌细胞钾离子通道蛋白活化水平方法的优化

介绍了一种如何合理的利用蛋白质免疫沉淀和蛋白质免疫印迹相结合的方法检测大鼠心肌细胞钾离子通道蛋白Kv1.2和Kv1.5的表达与活化水平.实验结果表明,与单独利用免疫印迹的方法相比较,本实验是对钾离子通道蛋白及其它亚家族的钾通道蛋白磷酸化表达水平检测方法的一种优化,从而获得一套可行、简单、合理的实验方案,同时也提高了检测的准确性,敏感性及特异性.     

The effects of severe carbon limitation on the green seaweed,Ulva conglobata (Chlorophyta)

Low inorganic carbon (Ci) concentrations in seawater are usually an important factor controlling photosynthesis and growth of seaweeds. The green seaweed, Ulva conglobata Kjellm, collected from a rock pool in a middle intertidal zone located at Nanao Island, Shantou, China, were cultured under low Ci level for several days, to examine the effect of severe carbon limitation on photosynthesis. The rather high pH compensation points obtained from the pH-drift experiments indicated that U. conglobata was capable of acquiring HCO₃ ^? from surrounding seawater as its Ci source for photosynthesis. However, thalli of U. conglobata cultured in Ci-starved seawater exhibited a decline of biomass, showing that the realistic photosynthetic carbon gain could not compensate for the respiratory carbon consumption in the thalli under severe Ci limitation during laboratory culture. Compared with ambient Ci conditions, the culture under severe Ci limitation significantly had an increased pigment content, but a lower maximum quantum yield and photosynthetic electron transport rate. Additionally, the maximum carbon-saturating photosynthesis rate and the apparent photosynthetic conductance of U. conglobata thalli increased in cultures with severe Ci limitation compared with ambient Ci in low N-grown thalli. The results suggest that under severe Ci limitation, U. conglobata thalli increased capacities of both light absorption processes and carbon fixation pathways.     

应用寡核苷酸微阵列检测肺癌样品中的P53和K-ras基因点突变

对影响寡核苷酸微阵列检测点突变的敏感性和特异性的各种因素,如杂交液,杂交温度,标记引物浓度及其比例等,进行了研究,采用不对称PCR扩增有利于敏感性提高,多重不对称PCR不影响杂交的特异性,且敏感性有所增加,对30例肺癌标本进行寡核苷酸微阵列检测,发现12例标本发生了P53基因来点突变,K-ras突变有5例,与测序结果相比,P53基因突变符合率达到80％,由于检测样本较少且检测位点不完全,因而未得到K－ras和P53基因突变与肿瘤的种类,病期及吸烟之间的明显相关性。     

猪O型口蹄疫病毒抗体化学发光酶联免疫检测方法的建立

表达并纯化猪O型口蹄疫病毒(FMDV)VP1重组蛋白作为检测抗原,建立了一种快速检测猪O型口蹄疫病毒抗体的化学发光酶联免疫(CLEIA)检测方法。建立的VP1-CLEIA方法特异性为100%,板内变异系数在1.10%–6.70%之间,板间变异系数在0.66%–4.80%之间,具有较好的特异性和重复性,且灵敏度高于ELISA方法。通过对山东、辽宁、河北地区采集的250份临床血清的检测表明,该方法与间接ELISA试剂盒的符合率为93.50%,与液相阻断ELISA试剂盒的符合率为94.00%,表明本次建立的VP1-CLEIA检测方法可以用于猪O型FMDV感染或疫苗免疫后抗体水平检测。     

Is it necessary for screening irregular antibodies of blood donors?

To explore the necessity of electronic crossmatching applied to irregular antibodies from blood donors, to ensure blood transfusion safety. Irregular antibody screening was performed on blood samples collected in the Dongguan Blood Center from Apr 17, 2014 to Dec 31, 2017. Primary screening was performed on the Sanquin automatic blood group analyzer by the microcolumn gel method (Sanquin). The positive samples were analyzed again using the salt medium method, polybrene method and micro-column gel method (Diana) for the second screening. If the second screening was positive, it was used to determine irregular antibody specificity (using panel cells) and any irregular antibody titer was detected. A total of 208,004 samples were detected, of which 316 were positive (316/208,004; 0.152%). Among them, 120 alloantibodies (120/135,139; 0.088%) were detected in male donors, which was much lower than in female donors (173/72,865; 0.237%) (P<0.01). A total of 16 kinds of known irregular antibodies and 40 cases of unknown antibody specificity were detected, with 119 cases of IgG type and 197 cases of IgM type, at the ratio of 1.65:1. In female donors, the frequencies of anti-D, anti-E, anti-M and anti-Le^a were significantly higher than in male donors (P<0.01). In married couples, the frequencies of anti-D and anti-E were significantly higher than those unmarried (P<0.05). In minority nationalities, the frequency of anti-M was significantly higher than in the Han nationality (P<0.05). In non-Guangdong donors, the frequencies of anti-D and anti-Le^a were significantly higher than in Guangdong donors. 87.02% of irregular antibody positive donors’ antibody titers were lower than “++”, which was deemed as no serious hazard for clinical transfusion. The study suggests that it is unnecessary to screen for irregular antibodies in blood donors. Male donors from Guangdong may not be required to undergo screening for irregular antibodies, and anti-D and anti-E only identification is also not required to be detected in female donors.     

TNNI3K,a Cardiac-Specific Kinase,Promotes Physiological Cardiac Hypertrophy in Transgenic Mice

Purpose

Protein kinase plays an essential role in controlling cardiac growth and hypertrophic remodeling. The cardiac troponin I-interacting kinase (TNNI3K), a novel cardiac specific kinase, is associated with cardiomyocyte hypertrophy. However, the precise function of TNNI3K in regulating cardiac remodeling has remained controversial.

Methods and Results

In a rat model of cardiac hypertrophy generated by transverse aortic constriction, myocardial TNNI3K expression was significantly increased by 1.62 folds (P<0.05) after constriction for 15 days. To investigate the role of TNNI3K in cardiac hypertrophy, we generated transgenic mouse lines with overexpression of human TNNI3K specifically in the heart. At the age of 3 months, the high-copy-number TNNI3K transgenic mice demonstrated a phenotype of concentric hypertrophy with increased heart weight normalized to body weight (1.31 fold, P<0.01). Echocardiography and non-invasive hemodynamic assessments showed enhanced cardiac function. No necrosis or myocyte disarray was observed in the heart of TNNI3K transgenic mice. This concentric hypertrophy maintained up to 12 months of age without cardiac dysfunction. The phospho amino acid analysis revealed that TNNI3K is a protein-tyrosine kinase. The yeast two-hybrid screen and co-immunoprecipitation assay identified cTnI as a target for TNNI3K. Moreover, TNNI3K overexpression induced cTnI phosphorylation at Ser22/Ser23 in vivo and in vitro, suggesting that TNNI3K is a novel upstream regulator for cTnI phosphorylation.

Conclusion

TNNI3K promotes a concentric hypertrophy with enhancement of cardiac function via regulating the phosphorylation of cTnI. TNNI3K could be a potential therapeutic target for preventing from heart failure.     

棉田动物群落动态及天敌与害虫种群之间的关系

通过对棉田动物群落动态及天敌与害虫种群之间关系的系统研究,得出：l）棉田生物多样性指数越大,群落稳定性越高;2）龟纹瓢虫与棉蚜种群数量之间极相关,草间小黑株与棉蚜种群之间有相关关系,棉蚜及龟纹瓢虫种群增长为指数增长;3）棉蚜及其多种天敌在水平分布上均为聚集格局;4）棉蚜、龟纹瓢虫及草间小黑蛛在垂直分布上均为聚集格局,龟纹瓢虫对棉蚜聚集场所具有追随跟踪攻击作用．