Moxibustion inhibits interleukin-12 and tumor necrosis factor alpha and modulates intestinal flora in rat with ulcerative colitis

AIM: To investigate the effect of moxibustion on intestinal flora and release of interleukin-12 (IL-12) and tumor necrosis factor-α (TNF-α) from the colon in rat with ulcerative colitis (UC). METHODS: A rat model of UC was established by local stimulation of the intestine with supernatant from colonic contents harvested from human UC patients. A total of 40 male Sprague-Dawley rats were randomly divided into the following groups: normal (sham), model (UC), herb-partition moxibustion (HPM-treated), and positive control sulfasalazine (SA-treated). Rats treated with HPM received HPM at acupuncture points ST25 and RN6, once a day for 15 min, for a total of 8 d. Rats in the SA group were perfused with SA twice a day for 8 d. The colonic histopathology was observed by hematoxylin-eosin. The levels of intestinal flora, including Bifidobacterium, Lactobacillus, Escherichia coli (E. coli), and Bacteroides fragilis (B. fragilis), were tested by real-time quantitative polymerase chain reaction to detect bacterial 16S rRNA/DNA in order to determine DNA copy numbers of each specific species. Immunohistochemical assays were used to observe the expression of TNF-α and IL-12 in the rat colons. RESULTS: HPM treatment inhibited immunopathology in colonic tissues of UC rats; the general morphological score and the immunopathological score were significantly decreased in the HPM and SA groups compared with the model group [3.5 (2.0-4.0), 3.0 (1.5-3.5) vs 6.0 (5.5-7.0), P < 0.05 for the general morphological score, and 3.00 (2.00-3.50), 3.00 (2.50-3.50) vs 5.00 (4.50-5.50), P < 0.01 for the immunopathological score]. As measured by DNA copy number, we found that Bifidobacterium and Lactobacillus, which are associated with a healthy colon, were significantly higher in the HPM and SA groups than in the model group (1.395 ± 1.339, 1.461 ± 1.152 vs 0.045 ± 0.036, P < 0.01 for Bifidobacterium, and 0.395 ± 0.325, 0.851 ± 0.651 vs 0.0015 ± 0.0014, P < 0.01 for Lactobacillus). On the other hand, E. coli and B. fragilis, which are associated with an inflamed colon, were significantly lower in the HPM and SA groups than in the model group (0.244 ± 0.107, 0.628 ± 0.257 vs 1.691 ± 0.683, P < 0.01 for E. coli, and 0.351 ± 0.181, 0.416 ± 0.329 vs 1.285 ± 1.039, P < 0.01 for B. fragilis). The expression of TNF-α and IL-12 was decreased after HPM and SA treatment as compared to UC model alone (4970.81 ± 959.78, 6635.45 ± 1135.16 vs 12333.81 ± 680.79, P < 0.01 for TNF-α, and 5528.75 ± 1245.72, 7477.38 ± 1259.16 vs 12550.29 ± 1973.30, P < 0.01 for IL-12). CONCLUSION: HPM treatment can regulate intestinal flora and inhibit the expression of TNF-α and IL-12 in the colon tissues of UC rats, indicating that HPM can improve colonic immune response.

蜂王浆主蛋白对人胚肺成纤维细胞HFL-I的抗衰老作用及其作用机制研究（英文）

目的:探讨不同浓度蜂王浆主蛋白(MRJPs)对人胚肺成纤维细胞HFL-I的抗衰老作用,并对其可能的分子机理进行预测。创新点:本实验采用拉曼光谱法探究添加MRJPs对HFL-I细胞内成分的影响。实验发现添加MRJPs的HFL-I与对照组的HFL-I的拉曼光谱图中,与DNA和蛋白质合成相关的拉曼峰发生了变化。该发现在未来可作为鉴定衰老HFL-I的一种手段。方法:实验以蜂王浆为原料,利用透析法得到MRJPs。通过对不同浓度MRJPs处理的HFL-I进行细胞形态学观察,测定HFL-I生命周期以及检测其β-半乳糖苷酶活性,评价了MRJPs的抗衰老效果。并且通过MTT法考察了不同浓度MRJPs对HFL-I的促增殖作用。同时,实验一方面采用实时定量聚合酶链反应(q PCR)测量了不同浓度MRJPs处理的HFL-I的细胞端粒相对长度以及MTOR、SOD1、CTNNB1、TP53四种衰老相关基因的相对表达量;另一方面还通过拉曼光谱实验法,进一步探索了MRJPs抗衰老的分子机理。结论:本实验结果显示,添加MRJPs能降低HFL-I衰老细胞所占比例,显著延长HFL-I的生命周期,一定量的MRJPs还能够促进HFL-I的增殖。此外,MRJPs还能减缓细胞端粒长度的缩短,上调衰老相关基因SOD1的表达,下调抗衰老基因MTOR、CTNNB1和TP53的表达。通过MRJPs添加组细胞的拉曼光谱上出现的独特的碱基、酰胺羰基和芳香氢峰,推测MRJPs的抗衰老作用可能与促进DNA及蛋白质在细胞中的合成密切相关。

山茶油对大鼠脂肪肝作用的新观察

目的:评价山茶油、大豆油和橄榄油对非酒精性脂肪性肝(NAFLD)病的影响。创新点:通过不同食用油的脂肪酸成分及对大鼠血糖、血脂、肝脏组织细胞特性和超微结构影响的比较分析,揭示了山茶油对NAFLD的影响与n-6/n-3比率不完全相关,可能还涉及其它因子。方法:分析了山茶油、大豆油和橄榄油的脂肪酸成分;测定和分析了三种油对大鼠的体重、肝脏系数、血脂的影响;采用细胞学和透射电镜技术对大鼠肝进行细胞学和亚细胞结构的观察。结论:山茶油、橄榄油和大豆油的多不饱和脂肪酸n-6/n-3比率分别为7.69%、12.50%和33.33%,但三种油对大鼠血糖、血脂、NAFLD肝细胞特性和亚细胞结构的影响结果比较表明,山茶油对诱导大鼠NAFLD的风险性高于大豆油,但低于橄榄油。这为油脂饮食对NAFLD的影响提供了新的见解。

饲料中添加姜黄渣对高温环境下养殖中华鳖存活率的改善作用（英文）

目的:评估饲料中添加姜黄渣对中华鳖存活率及抗高温应激能力的改善作用;通过中华鳖的抗氧化能力分析,探讨姜黄渣的生化作用机制。创新点:证实了饲料中添加姜黄渣对中华鳖具有增强抗高温能力,提高存活率的作用,为姜黄渣作为新的水产饲料添加剂的资源开发应用提供了重要科学依据。方法:将320只雌性中华鳖(体重约250 g)随机分为对照组(标准饲料)和实验组(饲料中添加10%的姜黄渣)。在2013年7月到10月分别考察实验组和对照组中华鳖的体重、存活率;分析试验地海盐县的气温变化与中华鳖存活率的相关性;用超氧化物歧化酶(SOD)和丙二醛(MDA)试剂盒分别检测了两组中华鳖肝组织的SOD酶活力及MDA含量;用高效液相色谱分别测定比较了两组中华鳖皮肤的姜黄素含量。结论:补充姜黄渣的中华鳖皮肤呈现金黄色,皮肤所含色素为姜黄素,平均含量为(0.14±0.03)μg/g。存活率调查显示,实验组的中华鳖存活率比对照组高135.5%;SOD和MDA分析显示,实验组中华鳖肝组织的SOD酶活比对照组高112.8%,MDA含量比对照组低36.4%。根据气温资料分析推测,持续高温容易导致中华鳖死亡,补充姜黄渣可有效降低高温造成的死亡率。综合分析显示,补充姜黄渣能显著提高中华鳖的存活率及抗高温应激能力,其作用与姜黄渣所含姜黄素的抗氧化活性相关。

用高度特异性王浆主蛋白1多克隆抗体ELISA法检测蜂王浆新鲜度(英文)

目的：为蜂王浆主蛋白1（MRJP1）的快速检测和鉴别提供科学依据,为蜂王浆的质量控制提供技术支持。创新点：首次比较了MRJP1特异性多克隆抗体与MRJP1重组表达蛋白多克隆抗体对王浆主蛋白家族的免疫反应差异,验证了蜂王浆中MRJP1蛋白降解与保温时间的相关性,建立了以MRJP1作为蜂王浆新鲜度生物标志物的快速检测方法。方法：通过蜂王浆主蛋白家族蛋白的氨基酸序列同源性分析,筛选出MRJP1的特异性多肽区域,进行人工合成,免疫兔子后取血清制备成特异性多克隆抗体。用蛋白质印迹法（Western blot）检测了MRJP1特异性多克隆抗体与MRJP1重组表达蛋白多克隆抗体对王浆主蛋白家族的免疫反应。以新鲜蜂王浆为对照品,用MRJP1特异性抗体酶联接免疫吸附剂测定（ELISA）法和变性电泳胶灰度扫描法分别测定保温（40°C）7-49天的蜂王浆中MRJP1含量的变化,并进行了相关性分析。结论：MRJP1的特异性抗体对MRJP1蛋白具有专一的免疫识别特性,可特异性地检测代表蜂王浆新鲜度的MRJP1含量变化,并鉴别蜂王浆的真伪。

Expression of a bee venom phospholipase A_2 from Apis cerana cerana in the baculovirus-insect cell

Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2) gene from the Chinese honeybee(Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector.Tn-5B-4(Tn) cells were transfected with the recombinant bacmid DNA for expression.Sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa.Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins.The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2(BvPLA2 of the European honeybee,Apis mellifera) polyclonal serum.The reaction resulted in a double glycosylation band,which agrees with the band generated by the native AmPLA2 in Western blot analysis.The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg).In summary,the recombinant AccPLA2 protein,a native BvPLA2-like structure with corresponding biological activities,can be glycosylated in Tn cells.These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.

一种基于频率细分的X射线脉冲星周期估计方法（英文）

目的:针对现有脉冲星周期估计算法精度低的问题,研究一种利用短时观测数据进行高精度X射线脉冲星周期估计的算法,为促进实时高精度X射线脉冲星导航提供算法支撑。创新点:提出频率细分的方法,推导continuous Lomb periodogram(CLP),实现X射线脉冲星非等间隔到达光子序列在细分频率处的频域分析。该方法可以显著减少运算复杂度,同时提高频域分析的频率分辨率,进而提高脉冲星周期估计的精度。方法:首先,考虑到X射线脉冲星信号是非等间隔到达的光子序列,本文采用专门用于非等间隔数据处理的fast Lomb方法对一段短时观测的脉冲星实测数据进行频域分析,获得一个初始频率作为脉冲星旋转频率的初值。然后,在该初始频率附近以高的频率分辨率做频率细分,获取设定数量的高精度细分频率。最后,对该段短时观测的脉冲星数据在这些细分频率处做CLP分析。在CLP中,峰值位置所对应的频率即为估计出的具有较高精度的脉冲星旋转频率,由该频率就可以确定高精度的脉冲星旋转周期。实测数据分析表明:当观测数据小于135 s时,本文算法的周期估计精度比fast Lomb方法和FFT方法高1到3个数量级,且仅增加了一点计算复杂度。同时,相比于HEAsoft的周期估计方法(efsearch),本文算法具有精度高计算复杂度低的优势。结论:本文算法解决了fast Lomb方法在周期估计时精度受数据长度和观测时间限制的问题,显著提高了X射线脉冲星周期估计的精度并降低了计算复杂度。同时短时高精度的周期估计有助于提高TOA估计精度及X射线脉冲星导航中实时位置和速度的估计精度。本文算法还可以用于变星及其他天体的周期估计。

蜂王浆主蛋白(MRJPs)作为灭活胎牛血清(FBS)替代品培养人体细胞的效果评价（英文）

目的:为MRJPs应用于人体细胞培养提供科学依据,为发展蜂王浆在细胞工程的新用途提供技术支撑。创新点:首次验证MRJPs对多种人体细胞的促分裂效果,证实MRJPs可部分替代FBS培养人体细胞,并取得培养人体细胞的MRJPs与FBS和细胞因子的适合比例。方法:鲜蜂王浆中分离得到的MRJPs溶液与FBS按不同比例(0/10、3/7、6/4和9/1)复配成复合血清,分别添加于无血清培养基中用于培养293T、HFL-I、231、HCT116和Changliver等5种人体细胞。以添加体积分数为10%的FBS的无血清培养基为对照,根据MTT法测定的吸光度和细胞形态比较分析结果,得到促进细胞分裂的MRJPs溶液与FBS最佳配合比例。然后在筛选出的最佳复合血清中添加不同的细胞因子,再通过同样的比较分析,得到适合的细胞因子组合。结论:MRJPs对多种人体细胞具有促进分裂作用,可部分替代FBS培养人体细胞。MRJPs与FBS的最佳配比为:60%MRJPs溶液和40%FBS(复合血清);该复合血清与细胞因子的最佳组合为:复合血清+表皮生长因子+胰岛素-转铁蛋白-硒。