Ethanol extract from Cortex Albiziae attenuates mouse acute liver injury induced by carbon tetrachloride via modulation of NF-κB pathway

AIM:To investigate the protective effect of ethanol extract from Cortex Albiziae on acute liver injury, and to explore its possible mechanism. METHODS:Acute liver injury in mice was induced by single intraperitoneal injection of 25% carbon tetrachloride (olive oil solubilization). The effective parts of ethanol extract from Cortex Albizziae against acute liver injury were screened. The pathological changes of the liver tissues were examined by pathological sections with HE staining. The activity of total superoxide dismutase (T-SOD) and the content of malondialdehyde (MDA) of the liver tissues were detected, the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were mea-sured by ELISA, and the protein expression levels of NF-κB p65, Bcl-2 and Bax in the liver cells of the mice in each group were determined by Western blot. RESULTS:Compared with model group, the serum levels of AST and ALT in low-dose n-butanol phase of ethanol extract from Cortex Albiziae (AB-L, 4 mg·kg^-1·d^-1) group and high-dose n-butanol phase of ethanol extract from Cortex Albiziae (AB-H, 8 mg·kg^-1·d^-1) group were significantly decreased. The necrosis extent and degree of the hepatocytes and infiltration of inflammatory cells were significantly lower than that in model group. Compared with model group, the serum levels of TNF-α and IL-6 in AB-H group and AB-L group were significantly decreased (P<0.05). The protein level of NF-κB p65 in the nuclei of mouse liver cells in AB-H group and AB-L group were also decreased significantly (P<0.05). Compared with model group, the protein expression of Bax was decreased, the protein expression of Bcl-2 was increased, and the Bcl-2/Bax ratio was increased in AB-L group and AB-H group. CONCLUSION:The n-butanol phase of ethanol extract from Cortex Albiziae may protect the liver by reducing the activation of NF-κB p65, inhibiting the excessive release of inflammatory cytokines IL-6 and TNF-α, and decreasing hepatocyte apoptosis via regulating Bcl-2 and Bax expression.     

miR-204 inhibits proliferation of Hodgkin lymphoma cells by targeting Sirt1/p53 signaling pathway

AIM: To investigate the effect of microRNA-204 (miR-204) on the proliferation of Hodgkin lymphoma cells and the underlying mechanism. METHODS: The expression of miR-204 and Sirt1 mRNA in Hodgkin lymphoma tissues was detected by RT-qPCR. After transfection with miR-204 mimic, Sirt1 siRNA and miR-204 mimic+pcDNA3.1-Sirt1 into the L428 cells, the cell viability and BrdU incorporation were measured by CCK-8 assay and BrdU assay, respectively. The protein levels of Sirt1 and acetylated p53 (ac-p53) were determined by Western blot.The targeting relationship between miR-204 and Sirt1 was verified by double luciferase reporter assay. RESULTS: The low expression of miR-204 and the high mRNA expression of Sirt1 were found in the Hodgkin lymphoma tissues. Compared with control group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were significantly decreased after L428 cells were transfected with miR-204 mimic or Sirt1 siRNA (P<0.05). Compared with miR-204 mimic alone group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were increased after L428 cells were co-transfected with miR-204 mimic and pcDNA3.1-Sirt1 (P<0.05). The results of double luciferase reporter assay confiermed that Sirt1 was the target gene of miR-204. CONCLUSION: The inhibitory effect of miR-204 on the proliferation of L428 cells may be achieved by inhibiting the expression of Sirt1 and promoting the up-regulation of ac-p53.     

黑麦草（Lolium perenne）根际细菌多样性对Ni污染沉积物修复效果的响应

应用黑麦草（Lolium perenne）对某滨海城市南排污河的重金属污染沉积物——底泥进行植物修复,并结合变性梯度凝胶电泳（DGGE）分析了黑麦草根际土壤的重金属Ni含量和细菌多样性的关系。结果表明,黑麦草对Ni的修复效果较好,种植黑麦草后根际土壤Ni浓度降低了11.8%。根际土壤中Ni有效态含量与根际土壤pH值的变化呈较好的正相关关系,相关系数为0.968 8。对根际土壤细菌多样性进行分析,重金属Ni含量变化能够影响土壤细菌的多样性,根际土壤细菌多样性随黑麦草的生长而不断演变,且与根际土壤中Ni浓度变化有密切联系。对DGGE电泳后回收的部分片段进行测序并建立了进化树,结果表明,测序片段与非培养方法得到的菌种的同源性较高,且其均在土壤或河道底泥的相关研究中被发现。不同时期细菌群体组成和Shannon指数随着修复时间而变化,并与细菌总数有正相关关系。     

鸡p15INK4B基因的克隆表达及纯化

为研究鸡p15INK4B蛋白的功能及为制备此蛋白的单克隆抗体提供材料,应用引物搭桥法合成p15INK4B基因,应用基因工程的手段构建p15INK4B基因的原核表达载体,在E.coliRosetta(DE3)菌进行融合表达,并对表达产物进行亲和层析纯化。结果成功地构建了p15INK4B基因原核表达载体,经亲和层析制备了较高纯度的目的表达产物。     

牛瑟氏泰勒虫双拷贝p23表面蛋白基因真核表达

为构建牛瑟氏泰勒虫双拷贝p23表面蛋白基因真核表达质粒,根据GenBank牛瑟氏泰勒虫p23表面蛋白基因序列（D84447）,分别设计2对特异性引物,利用全血基因组DNA提取试剂盒提取牛瑟氏泰勒虫基因组DNA,采用SOE—PCR技术构建双拷贝p23基因,克隆到pMD-18-T载体上,经过PCR、酶切鉴定及测序后,亚克隆到pVAX-Ⅰ真核表达载体上,经过鉴定后采用脂质体法将重组质粒pVAXI-2p23转染到BHK-21细胞,用IFA和RT—PCR来鉴定目的基因的表达情况．结果表明,成功构建了牛瑟氏泰勒虫双拷贝p23表面蛋白基因真核表达质粒,并在BHK-21细胞中获得表达．     

绵羊肺炎支原体 p113 基因的序列分析及功能预测

采用多种软件对p113基因的相似性、跨膜结构、重复序列、信号肽、抗原表位等进行预测,并与同源序列进行比较分析.结果表明,p113基因是猪肺炎支原体黏附素基因p97的直系同源基因,并具有与P97蛋白R2区类似的特征性重复序列.通过综合比较分析,推测P113蛋白极可能是绵羊肺炎支原体的黏附素和膜表面免疫原.     

水稻启动子OsN1p的克隆与功能分析

将OsN1基因上游881 bp启动子(OsN1p)序列取代pBI121中gus基因上游的35S启动子,构建植物表达载体pBIN1p,经农杆菌介导转化水稻品种‘日本晴’,获得转基因植株。GUS组织化学染色结果表明,由该启动子驱动的gus基因能在愈伤组织中较低水平地表达;稻瘟病菌接种转基因植株24 h后,GUS活性为未接种前的4.2倍;5 mmol.L-1水杨酸(SA)和0.5 mmol.L-1茉莉酸甲酯(MeJA)分别喷施转基因植株叶面6 h后,GUS活性分别为处理前的5.9和2.4倍。表明,OsN1p启动子具有启动活性,同时明显具有受稻瘟病菌、SA和MeJA诱导表达的特性。     

多因子对墨吉明对虾氮磷代谢的影响

对不同条件下的墨吉明对虾氮磷代谢进行了相关实验。实验结果表明：温度、体质量、盐度、p H、摄食状态和光照条件都对墨吉明对虾磷、硝酸氮、亚硝酸氮和氨氮代谢影响显著。在20~30℃范围内,随温度升高,墨吉明对虾磷、亚硝酸氮、氨氮代谢率均显著升高;硝酸氮代谢率在25℃条件下最大;磷、亚硝酸氮、硝酸氮代谢率随体质量增加而减慢,氨氮代谢加快;盐度在10~30范围内,随盐度升高,磷、亚硝酸氮代谢率提高,硝酸氮、氨氮代谢率下降;p H在7.5~9.0范围内,随p H值升高,磷代谢水平明显下降,氨氮代谢率显著升高,亚硝酸氮代谢率在p H 8.5时出现最大值（0.451±0.039）μg/（g·h）;硝酸氮代谢率在p H 8.0时有最小值（1.364±0.028）μg/（g·h）;饱食状态下墨吉明对虾磷、硝酸氮、亚硝酸氮、氨氮代谢率相对于饥饿组分别提高了33.85%、68.07%、233.81%、72.22%;正常光照条件下墨吉明对虾磷、硝酸氮、亚硝酸氮、氨氮代谢率相对于遮光组分别提高了130.79%、37.58%、120.23%、103.93%。     

新型穿膜肽介导 p53融合蛋白的表达及其免疫原性分析

目的：原核表达新型细胞穿膜肽RDP介导p53融合蛋白,并检测其免疫原性的强弱．方法：以质粒pET28a‐p53为模板扩增p53基因,并克隆至原核表达载体pET28a‐RDP中,构建重组表达质粒pET28a‐RDP‐p53,转化至大肠杆菌Rosetta ,IPTG诱导表达蛋白并纯化,SDS‐PAGE确定该蛋白准确性．同时,将昆明小鼠分为空白对照组（等容生理盐水）和RDP‐p53融合蛋白高、中、低剂量（4 mg／kg ,2 mg／kg ,1 mg／kg）组,经腹腔注射免疫,每隔2 d给药1次,30 d后眼球取血,用酶联免疫吸附法（ELISA）测定血清中IgG的含量．结果：双酶切及测序结果显示, p53基因已克隆入表达载体中;RDP‐p53融合蛋白在上清和沉淀中均获得表达．ELISA测定结果表明,与对照组比较,低、中剂量组小鼠血清中IgG含量没有明显升高（p＞0.05）,高剂量组显著升高（p＜0.05）．结论：成功表达并纯化RD P‐p53融合蛋白,其免疫原性较弱且与给药剂量相关,为进一步研究RD P‐p53融合蛋白对脑肿瘤的药理作用奠定基础．