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1.
AIMTo investigate whether Rho-associated coiled-coil kinase (ROCK) is involved in high glucose-induced apoptosis of primary cardiomyocytes by regulating PI3K/Akt signaling pathway. METHODSPrimary Wistar rat cardiomyocytes were cultured and identified by α-sarcomeric actin (α-SCA) immunohistochemistry. Cardiomyocytes were treated with 5.5, 33 and 40 mmol/L glucose for 48 h. The cell viability was measured by MTT assay, and the mRNA expression of ROCK1 and ROCK2 in the cardiomyocytes was detected by RT-qPCR. Flow cytometry was used to analyze the apoptosis of the cardiomyocytes. The protein levels of ROCK1, ROCK2, cleaved caspase-3, Bcl-2, PI3K, Akt and p-Akt were determined by Western blot. In order to confirm the regulatory effect of ROCKs on PI3K/Akt signaling pathway, the cells were divided into control group (5.5 mmol/L glucose), high glucose group (33 mmol/L glucose) and high glucose+Y27632 (ROCK inhibitor) group. Western blot was used to detect the protein levels of ROCK1, ROCK2, PI3K, Akt and p-Akt. RESULTSAfter 48 h of high glucose exposure, the values of relative cell viability in 33 and 40 mmol/L glucose groups were (79.71±2.43)% and (68.41±7.49)%, respectively, both of which were significantly decreased compared with normal control group (P<0.05). After 48 h of high glucose exposure, the relative mRNA levels of ROCK1 and ROCK2 in 33 and 40 mmol/L glucose groups were significantly increased compared with normal control group (P<0.05). Compared with normal control group, the apoptotic rate in 33 and 40 mmol/L glucose groups was increased significantly (P<0.05). Compared with normal control group, the protein expression of ROCK1, ROCK2 and cleaved caspase-3 in 33 and 40 mmol/L glucose groups was increased (P<0.05), while the protein expression of Bcl-2 was decreased (P<0.05). No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed, while the protein level of p-Akt in 33 and 40 mmol/L glucose groups was decreased compared with normal control group (P<0.05). Compared with high glucose group, the expression of ROCK1 and ROCK2 was decreased in high glucose+Y27632 group. No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed. Compared with normal control group, the protein level of p-Akt in high glucose group was decreased, and the protein level of p-Akt in high glucose+Y27632 group was increased significantly compared with high glucose group. CONCLUSION Under high glucose environment, ROCK may reduce the level of p-Akt by inhibiting the PI3K/Akt signaling pathway, thus promoting the apoptosis of cardiomyocytes.  相似文献   
2.
A 10‐week feeding trial was conducted to evaluate the growth performance, glucose transport and metabolism of Chinese soft‐shelled turtles (Pelodiscus sinensis) exposure to graded levels of dietary starch (0.52%, 7.43%, 14.74%, 22.99% and 31.38%). The 360 turtles (initial body weight, 12.94 ± 0.50 g) with 12 replicates were randomly assigned to five experimental diets. The highest weight gain and specific growth rate (SGR) were observed in 7.43% group and the lowest in 31.38% group. The protein efficiency ratio, whole‐body lipid contents, hepatic glycogen contents and the 4‐hr postprandial plasma glucose levels were significantly increased with the increment of starch levels (p < .05). In contrast, the daily feed intake and feed conversion ration were significantly declined (p < .05). The mRNA levels of glucose transporter 2, glucokinase, pyruvate kinase, malic enzyme and acetyl‐CoA carboxylase alpha genes in the liver significantly increased as the increase in starch levels at 4‐hr and 24‐hr post feeding (p < .05). No significant differences were observed in the expression of gluconeogenesis genes at each time point (p > .05). These results suggested that dietary addition of starch up‐regulated hepatic glycolysis, glycogenesis and lipogenesis genes expression, but the deficient response of gluconeogenesis to dietary starch might be part of the causes limited the starch utilization. Based on the secondary polynomial regression of SGR, y = ?0.0011x2 + 0.028x + 1.63 (R2 = 0.9292), the 12.73% inclusion level of dietary starch was recommended in juvenile turtles.  相似文献   
3.
研究发现NH_2-MIL-101(Fe)金属有机框架材料可催化H_2O_2氧化3,3,5,5-四甲基联苯胺(TMB)显蓝色,表现出过氧化物模拟酶特性,而且在较宽的温度(4~80℃)及pH值范围(2~10)内保持其模拟酶活性,结合葡萄糖氧化酶,建立了测定葡萄糖的方法.在优化条件下,吸光度与葡萄糖浓度在0.75~50μmol/L范围内呈现良好的线性关系,对葡萄糖的检出限为0.75μmol/L.将本法用于血清中葡萄糖的测定,获得满意结果.  相似文献   
4.
Skin mucus has been demonstrated to provide stress biomarkers for evaluating the physiological status, providing new convenient and non‐invasive methods to detect stress response in fish. Here, we investigated the anaesthetic efficacy of tricaine methanesulphonate (MS‐222; 75–115 mg/L) for discus Symphysodon aequifasciata (34.27 ± 4.46 g; 8.10 ± 0.59 cm) using skin mucus stress biomarkers. The induction time, recovery time and respiratory frequency were also determined. According to the criteria for anaesthesia and recovery, discus fish to reach stage A3 (deep anaesthesia) within 3 min and to reach stage R4 (full recovery of normal behaviour) within 5 min were observed at 95–105 mg/L MS‐222. Respiratory frequency increased first and then decreased during MS‐222 exposure and increased after recovery. At 10 min after deep anaesthesia, a lower mucus glucose was only observed at 115 mg/L MS‐222. No change in mucus cortisol and increased lactate were observed in all treatments. Increased mucus protein was observed at 75, 85 and 95 mg/L MS‐222. At 10 min after recovery, increased mucus glucose and decreased mucus protein were observed at 85, 95 and 115 mg/L MS‐222, but increased mucus cortisol only at 115 mg/L and lactate only at 75 and 105 mg/L MS‐222. At 24 hr after recovery, mucus glucose returned to the initial level only at 75, 95 and 105 mg/L MS‐222, while cortisol at 75 and 85 mg/L and protein and lactate at 75 mg/L respectively. Overall, the effective dose of MS‐222 for discus fish has been suggested to be 95–105 mg/L.  相似文献   
5.
In dairy cows, glucose is essential as energy source and substrate for milk constituents. The objective of this study was to investigate effects of long‐term manipulated glucose and insulin concentrations in combination with a LPS‐induced mastitis on mRNA abundance of glucose transporters and factors involved in milk composition. Focusing on direct effects of insulin and glucose without influence of periparturient endocrine adaptations, 18 dairy cows (28 ± 6 weeks of lactation) were randomly assigned to one of three infusion treatments for 56 h (six animals each). Treatments included a hyperinsulinemic hypoglycaemic clamp (HypoG), a hyperinsulinemic euglycaemic clamp (EuG) and a control group (NaCl). After 48 h of infusions, an intramammary challenge with LPS from E. coli was performed and infusions continued for additional 8 h. Mammary gland biopsies were taken before, at 48 (before LPS challenge) and at 56 h (after LPS challenge) of infusion, and mRNA abundance of genes involved in mammary gland metabolism was measured by RT‐qPCR. During the 48 h of infusions, mRNA abundance of glucose transporters GLUT1, 3, 4, 8, 12, SGLT1, 2) was not affected in HypoG, while they were downregulated in EuG. The mRNA abundance of alpha‐lactalbumin, insulin‐induced gene 1, κ‐casein and acetyl‐CoA carboxylase was downregulated in HypoG, but not affected in EuG. Contrary during the intramammary LPS challenge, most of the glucose transporters were downregulated in NaCl and HypoG, but not in EuG. The mRNA abundance of glucose transporters in the mammary gland seems not to be affected by a shortage of glucose, while enzymes and milk constituents directly depending on glucose as a substrate are immediately downregulated. During LPS‐induced mastitis in combination with hypoglycaemia, mammary gland metabolism was more aligned to save glucose for the immune system compared to a situation without limited glucose availability during EuG.  相似文献   
6.
This study investigated the effect of repeated acute restraint stress and high‐fat diet (HFD) on intestinal expression of nutrient transporters, concomitant to intestinal inflammation. The ability of adenosine to reverse any change was examined. Six‐week‐old male Sprague Dawley rats were divided into eight groups: control or non‐stressed (C), rats exposed to restraint stress for 6 h per day for 14 days (S), control rats fed with HFD (CHF) and restraint‐stressed rats fed with HFD (SHF); four additional groups received the same treatments and were also given 50 mg/l adenosine dissolved in drinking water. Fasting blood glucose, plasma insulin, adiponectin and corticosterone were measured. Intestinal expression of SLC5A1, SLC2A2, NPC1L1 and TNF‐α was analysed. Histological evaluation was conducted to observe for morphological and anatomical changes in the intestinal tissues. Results showed that HFD feeding increased glucose and insulin levels, and repeated acute restraint stress raised the corticosterone level by 22%. Exposure to both stress and HFD caused a further increase in corticosterone to 41%, while decreasing plasma adiponectin level. Restraint stress altered intestinal expression of SLC5A1, SLC2A2 and NPC1L1. These changes were enhanced in SHF rats. Adenosine was found to alleviate HFD‐induced increase in glucose and insulin levels, suppress elevation of corticosterone in S rats and improve the altered nutrient transporters expression profiles. It also prevented upregulation of TNF‐α in the intestine of SHF rats. In summary, a combination of stress and HFD exaggerated stress‐ and HFD‐induced pathophysiological changes in the intestine, and biochemical parameters related to obesity. Adenosine attenuated the elevation of corticosterone and altered expression of SLC5A1, NPC1L1 and TNF‐α.  相似文献   
7.
为提高红霉素发酵单位,考察了二甲基亚砜(DMSO)添加时间和体积分数对红霉素合成的影响,采用筛选自身代谢产物突变株的方法,将突变株进行常压室温等离子体(ARTP)诱变和耐受高浓度DMSO处理。结果显示,在36 h向发酵瓶中添加30000μg/m L红霉素,筛选得到化学效价比对照提高22.4%的菌株。DMSO最适添加时间为发酵48 h,最适添加剂量为0.2%,其可提高发酵单位10.1%。  相似文献   
8.
本试验旨在通过人工瘤胃体外培养,研究不同代谢葡萄糖水平下的绵羊瘤胃发酵特性、微生物蛋白质浓度和产气参数。采用单因子试验设计,共设计4个代谢葡萄糖水平[125(A)、138(B)、153(C)、168 g/kg(D)]。体外试验所用瘤胃液采自4只安装有永久性瘤胃瘘管的绵羊。分别于培养0、2、4、6、8、12、24 h采集2 mL培养液用于分析。结果表明:1)8~24 h培养液pH随着代谢葡萄糖水平的提高而出现显著或极显著下降(P0.05或P0.01);氨氮浓度在2 h时D组显著高于A组(P0.05),而在6 h时A组显著高于其他3组(P0.05)。2)D组6h培养液细菌蛋白浓度显著高于A组(P0.05);随着代谢葡萄糖水平的提高,培养液丙酸、丁酸、总挥发性脂肪酸的浓度总体呈升高趋势,乙酸/丙酸呈下降趋势。3)随着代谢葡萄糖水平的提高,理论最大产气量极显著降低(P0.01),达1/2理论最大产气量的时间极显著缩短(P0.01);潜在产气量无显著变化(P0.05),24 h产气量和产气速率常数分别显著或极显著下降和上升(P0.05或P0.01)。结果提示,提高代谢葡萄糖水平,可以提高丙酸、总挥发性脂肪酸的浓度,同时可为绵羊提供较多的生糖前体物质。  相似文献   
9.
10.
甜玉米是世界上重要的果蔬两用型作物之一。建立快速、无损检测甜玉米糖分含量的方法,对于甜玉米品质育种工作中的材料鉴定、筛选具有重要意义。选取了104份糖分含量变幅较大的甜玉米材料进行了近红外光谱分析,结合酶法测量的化学值,利用偏最小二乘法以及不同光谱处理和数学处理相结合,对全光谱波段进行了化学计量学的分析统计,构建了葡萄糖、果糖和蔗糖含量的近红外光谱定标模型。用外部验证样品集进行验证,对所建模型的实际预测能力进行检验。结果表明:最优光谱处理方式葡萄糖为加权多元散射校正,果糖和蔗糖的为标准正常化加去散射处理;最优的导数处理葡萄糖为2阶导数,果糖和蔗糖的为1阶和3阶导数。葡萄糖、果糖和蔗糖定标模型的交叉验证相关系数分别为0.646、0.645、0.820,交叉验证标准偏差分别为0.321、0.275、1.508;外部验证集葡萄糖、果糖和蔗糖的预测相关系数分别为0.593、0.780、0.891,表明所建立的果糖和蔗糖预测模型具有较好的预测性,可应用于甜玉米种质资源筛选,而葡萄糖模型的预测性较差,需继续完善。  相似文献   
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