朱顶红幼嫩花梗胚性愈伤组织诱导和高效植株再生

为建立高效的朱顶红植株再生和种苗繁育技术体系，以幼嫩花梗为外植体开展胚性愈伤组织诱导和植株再生研究。对2,4-D和噻苯隆（TDZ）浓度、外植体发育时期及大小进行了筛选，结果表明：将切片厚度为1 mm的幼嫩花梗外植体置于添加0.5 mg ? L-1 TDZ 和2.0 mg ? L-1 2,4-D的MS固体培养基上培养8周，胚性愈伤组织诱导率最高，达85.3%。将胚性愈伤组织转移至相同的培养基上，每月继代1次，平均每月增殖10.6倍。在不含任何生长调节剂的MS培养基上，胚性愈伤组织的植株再生率达98.0%，平均每块愈伤组织可再生出12.3个小植株。经过36次（3年）继代培养后，胚性愈伤组织的增殖和植株再生效率没有显著变化。幼苗移栽至温室驯化，成活率达97.5%。再生植株移栽到田间，没有发现明显的表型变异。对随机选择的再生植株和母株进行简单序列重复区间（ISSR）扩增分析，证实再生植株没有发生DNA水平变异。     

响应面试验优化霍山石斛原球茎增殖培养基

为可持续开发霍山石斛资源，采用响应面法优化霍山石斛原球茎增殖培养基。在单因素试验基础上，根据Box Behnken试验设计原理，采用三因素三水平的响应面分析法，以增殖率为响应值进行回归分析。结果表明，萘乙酸（简称NAA）及6 苄氨基嘌呤（简称6 BA）浓度对增殖率影响最为显著，6 BA与马铃薯两者间交互作用显著，适宜的NAA、6 BA及马铃薯浓度分别为0.05 mg·L-1、0.70 mg·L-1和185 g·L-1，此时增殖率为2 989.82%±205.55%，与模型预测值基本符合。响应面法优化霍山石斛原球茎增殖培养基切实可行。     

Liver of the fish Gymnotus inaequilabiatus and nematode larvae infection: Histochemical features and expression of proliferative cell nuclear antigen

Histopathological lesions due to third‐larval stage of nematode Brevimulticaecum sp. within the liver of a subpopulation of 31 Gymnotus inaequilabiatus from the Pantanal Region (Brazil) were studied with histochemical and immunohistochemical methods. In 93.5% of fish, livers harboured nematode larvae and the intensity of infection ranged from 8 to 293. In livers with highest number of larvae, the hepatic tissue was occupied primarily by the nematodes. Each larva was encircled by focal inflammatory granulomatous reaction. Within the thickness of the granuloma, three concentric layers were recognized: an inner layer of densely packed epithelioid cells, a middle layer of mast cells (MCs) entrapped in a thin fibroblast‐connective mesh and an outer layer of fibrous connective tissue with fibroblasts. Epithelioid cells and fibroblasts within the thickness of the granuloma wall were positive for proliferative cell nuclear antigen (PCNA). Moreover, several hepatocytes in infected liver were immunoreactive to PCNA. Occurrence of rodlet cells and MCs in parenchyma, in close proximity to the encysted nematode larvae and near the blood vessel of infected liver, was observed. Macrophage aggregates (MAs) were numerous within the granulomas and scattered in parenchyma of the infected liver. High quantity of haemosiderin was encountered in MAs and hepatocytes of infected liver.     

‘红国王’葡萄组织培养快速繁殖

为建立‘红国王’葡萄组培快繁体系,本试验研究了生长素、接种材料类型、培养条件对组培苗继代增殖与生根的影响。结果表明,0.5 mg/L IAA适合‘红国王’的继代增殖,繁殖系数为5.49,在培养基中附加0.3 mg/L NAA,愈伤组织少且根系粗壮;适合‘红国王’组培苗生长的蔗糖浓度为25~30 g/L,在该浓度下植株长势健壮,生根条数多;带半叶单芽茎段为‘红国王’适宜接种材料类型;接种后第1天至第10天暗培养,可促进‘红国王’根系发育;放置于LED灯复合光红∶蓝∶白=3∶1∶1下,可获得长势强、根系强壮的组培苗;继代间隔为40 d时,繁殖速度较快。     

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.     

蝴蝶兰‘大辣椒’组培快繁技术体系的优化

探索优化蝴蝶兰组培快繁技术体系，以蝴蝶兰‘大辣椒’半木质化花梗为试验材料，研究消毒剂、基础培养基、植物生长调节剂配比及浓度对蝴蝶兰不定芽诱导、增殖和生根过程的影响。结果表明:1）先用2% H₂O₂溶液处理15 min，再用10% NaClO溶液处理10 min，消毒效果良好，花梗污染率低，为26.6%；2）不定芽诱导最适培养基为花宝1号+花宝2号+ 6-BA 3 mg/L+NAA 0.05 mg/L，出芽率为78.9%；3）不定芽增殖最适培养基为花宝1号+6-BA 5 mg/L+NAA 0.05 mg/L，增殖系数为3.60；4）幼苗生根最适培养基为1/2MS+NAA 0.2mg/L，生根率为73.7%。本研究旨在优化‘大辣椒’品种的组培快繁技术体系，为进一步建立蝴蝶兰再生体系和工厂化生产提供技术支持。     

蛋白质激发子Hrip1对小麦黄矮病的诱抗作用

小麦黄矮病是由大麦黄矮病毒（barley yellow dwarf virus，BYDV）引起的一种小麦病毒病，其传播介体是小麦蚜虫，在小麦生产中造成巨大的经济损失。近年来，植物诱导抗性作为一种新兴的植物病虫害防治措施引起了广泛的关注。蛋白质激发子Hrip1可以激活多种植物的免疫防御反应诱导植物产生广谱抗性。本研究评价了Hrip1对小麦黄矮病的诱抗效果。用30 μg/mL的Hrip1溶液进行小麦浸种和幼苗喷雾，随后接种BYDV，接种后第14 d，Hrip1对小麦黄矮病控制效果在50%以上，接种后第21 d控制效果仍在30%以上。实时荧光定量PCR检测结果显示，在Hrip1处理的小麦幼苗体内，BYDV外壳蛋白mRNA的数量显著低于对照组；EPG结果显示，在Hrip1处理的小麦幼苗上，麦二叉蚜寻找叶片刺吸位点和韧皮部取食位点的时间增加。以上结果表明：Hrip1能够有效地抑制BYDV在小麦体内的增殖；影响传毒媒介麦二叉蚜的取食行为，抑制其传毒能力。此外，Hrip1处理小麦能有效缓解BYDV引起的叶片黄化和植株矮化的症状。因此，Hrip1可以作为生物诱导剂综合控制小麦黄矮病。     

Expression and function of circular RNA_0000231 in non-small-cell lung cancer

AIM: To investigate the expression and function of circular RNA_0000231 (circ_0000231) in non-small-cell lung cancer (NSCLC). METHODS: RT-qPCR was used to detect the expression of circ_0000231 in the NSCLC tissues and cell lines. circ_0000231 small interfering RNA (si-circ_0000231) or negative control siRNA of circ_0000231 (NC) was transfected into the NSCLC cells. The proliferation and apoptosis of the NSCLC cells were detected by CCK-8 assay, colony formation assay and flow cytometry, respectively. The expression of cyclin D1 (CCND1) and anti-apoptotic protein Bcl-2 were determined by RT-qPCR and Western blot. RESULTS: The expression of circ_0000231 in the NSCLC tissues and cell lines was significantly up-regulated compared with precancerous tissues and lung epithelial cells BEAS-2B (P<0.05). After transfection of NSCLC cells with si-circ_0000231, the cell viability, colony formation numbers were significantly decreased, and the apoptotic rate in si-circ_0000231 group was significantly increased as compared with NC group (P<0.01). In addition, the results of RT-qPCR and Western blot showed that transfection of si-circ_0000231 inhibited the expression of CCND1 and Bcl-2 (P<0.01). CONCLUSION: The expression of circ_0000231 is significantly increased in the NSCLC tissues and cells. Knock-down of circ_0000231 expression significantly inhibits the proliferation of NSCLC cells.     

猪lncRNA TCONS_00791383对骨骼肌卫星细胞增殖分化的影响

为了探究lncRNA TCONS_00791383对猪骨骼肌卫星细胞增殖和分化的影响。本研究利用qRT-PCR技术检测出生7 d内大白仔猪6种组织（心、脾、肺、肾、背肌和腿肌）以及猪骨骼肌卫星细胞增殖分化前后TCONS_00791383的表达水平；通过设计反义核苷酸（antisense oligonucleotides，ASO）片段在猪骨骼肌卫星细胞中对TCONS_00791383进行敲低，检测敲低TCONS_00791383之后增殖分化标志基因的表达量变化；通过trans （co-expression）对TCONS_00791383进行靶基因预测，使用DAVID对其进行GO富集和KEGG通路分析。结果显示，TCONS_00791383在猪心脏中表达量最高，在脾和肾组织中不表达。在骨骼肌卫星细胞从增殖到分化的过程中，TCONS_00791383的表达量逐渐上升，且在分化后30 h表达量达到最高。在使用ASO片段敲低TCONS_00791383之后，与对照组相比，在分化24 h，增殖标志基因Pax3、Pax7表达量显著或极显著降低（P<0.05，P<0.01），分化标志基因MyoG表达量极显著降低（P<0.01），在分化48 h，增殖标志基因Pax3表达量极显著降低（P<0.01），Pax7表达量显著降低（P<0.05），分化标志基因MyHC表达量显著降低（P<0.05）。预测得到的相关靶基因富集到AMPK、ATP等多个与骨骼肌卫星细胞增殖和分化过程相关的重要信号通路。本研究表明，lncRNA TCONS_00791383可能促进猪骨骼肌卫星细胞的增殖和分化。