原花青素A-1体内外对猪与小鼠T淋巴细胞亚群的影响

为研究从碎米花杜鹃叶中分离得到的化合物原花青素A-1（Proanthocyanidin A-1，简称PAA-1）的免疫增强活性，初步从细胞水平上探讨其免疫增强机制。体外试验采用流式细胞术，检测PAA-1体外对小鼠脾淋巴细胞T细胞亚群CD4和CD8单阳性和双阳性T淋巴细胞亚群百分率及CD4+/CD8+比值；体内试验通过饲喂不同剂量的PAA-1后，检测试验猪外周血淋巴细胞CD4和CD8单阳性以及CD4+/CD8+比值。结果证明，体外试验中同阴性对照组相比，PAA-1能单独或协同Con A提升具CD4+、CD8+及CD4+CD8+表型的T淋巴细胞亚群百分率，提高CD4+/CD8+的比值；体内试验中饲喂中、高剂量的PAA-1的试验猪外周血中具有CD4+、CD8+表型的T淋巴细胞的百分率及CD4+/CD8+的比值均有显著提高。说明PAA-1可通过增加辅助性T淋巴细胞和细胞毒性T淋巴细胞数量、促进T淋巴细胞的成熟以及增加CD4+/CD8+的比值发挥增强细胞免疫的作用，为其进一步开发为新型免疫增强剂提供试验依据。     

黑龙江地区鲤疱疹病毒3型的流行病学调查及其主要囊膜蛋白的抗原表位预测

2015-2016年对来自黑龙江地区46个养殖场送检的鲤鱼进行锦鲤疱疹病毒(Koi herpes virus,KHV)的分离鉴定,并应用DNAStar等软件对检出的阳性样本的主要囊膜蛋白(Major envelop protein,MEP)基因进行生物信息学分析。结果显示,KHV的检出率约为6.5%,3株阳性样本具有相同的MEP基因序列,称之为KHV-HLJ1516。MEP基因开放阅读框为771 bp,编码256个氨基酸,相对分子质量为28 280.58,等电点为8.40。对KHV-HLJ1516的MEP基因序列同源性分析表明,其与KHV-GZ11分离株仅有1个碱基不同,但并未导致氨基酸发生改变,碱基相似性为99%;而与HZ419分离株相比,KHV-HLJ1516的MEP基因有3个碱基发生突变,相应的氨基酸由Asn变为IIe。聚类分析结果显示,KHV-HLJ1516与KHV-GZ11处于同一分支,而HZ419位于另一分支。B细胞抗原表位预测结果显示,KHV-HLJ1516的MEP的抗原表位主要位于4~11,24~40,42~63,82~110,209~218,238~249氨基酸区段。     

Antioxidant and anti‐inflammatory activities of Pinus radiata bark extract in salmonid cell lines

A fish meal supply shortage is limiting aquaculture development. Currently, plant‐based proteins, such as soya bean meal, are being used as an alternative protein source, despite that such a diet can adversely affect fish, such as by inducing an inflammatory response. A possible solution is to include dietary additives in farm diets to counteract negative effects. One such solution originates from pine bark extracts, which present bioactive properties. In this study, the antioxidant and anti‐inflammatory properties of Pinus radiata bark extracts were evaluated for the first time in a salmonid cell line. This extract chemically demonstrated antioxidant activity through 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH = 58.4 ± 1.1%) and ferric ion reducing antioxidant power (FRAP = 575 ± 17 mgEqFe(II)·g extract^?1) assays. Additionally, the extract showed high flavonoid and phenolic compound contents. Up to 100 mg mL^?1, the P. radiata extract showed no cytotoxicity in the CHSE‐214 salmonid embryo cell line. Moreover, the antioxidant activity of the extract (50 μg mL^?1) was evaluated by a dichlorofluorescein (DCFH) assay in the SHK‐1 salmon cell line challenged with an oxidant stimulus (H₂O₂), showing 58.9% activity. The extract also protected DNA from oxidative damage, as observed through a comet assay. When assessing anti‐inflammatory properties in an in vitro inflammation model, the extract significantly reduced the relative expression of the pro‐inflammatory cytokines interleukin‐1β (IL‐1β), tumour necrosis factor‐α (TNF‐α) and interleukin‐8 (IL‐8) and of the inducible cyclooxygenase‐2 (COX‐2) enzyme. These results suggest a potential application of P. radiata bark extract in functional foods in aquaculture.     

大白菜游离小孢子培养技术高效体系的研究

以大白菜吉红82、#534DH 群体及#438 群体为试材，研究了利用细胞破碎仪机械提取小孢子与人工挤压提取小孢子对大白菜游离小孢子活力及提取量的影响。结果表明：机械提取替代人工挤压可在短时间内同时提取多份样品，并可快速调整小孢子的培养浓度。收集40 个适期花蕾于5 mL 离心管内，3 000 r·min-1 破碎10 s，将提取的小孢子定容于25 mL NLN-13 培养基，可确保小孢子浓度在适宜培养范围内，并成功诱导胚胎发生。     

FGF4在肺癌细胞体内和体外中表达差异的研究

FGF4已经被证明是癌基因,它涉及肿瘤的生长和转移,为了解FGF4的表达与肿瘤微环境的关系。我们利用FGF4抗体通过免疫组化对一名肺癌患者癌旁,癌组织,癌组织小鼠移植瘤,二次移植瘤以及原代培养的细胞爬片进行FGF4检测,探究其表达差异。通过对比癌组织与对照组（癌旁组织）,FGF4在癌巢中高表达;同样,移植瘤与二次移植瘤的癌巢中与癌旁组织比较,FGF4表达相对较高;但是将癌组织进行原代培养后,免疫组化检测细胞爬片FGF4,发现仅有5%±0.21%的肿瘤细胞表达FGF4,对照蛋白Cytokine作为肿瘤标记物,则在100%的肿瘤细胞中表达。研究提示免疫组化检测到FGF4在体内和体外表达不同,提示肿瘤细胞FGF4的表达与肿瘤微环境调控密切相关,肿瘤微环境对肿瘤细胞的FGF4的调控有着重要作用。     

水稻半外卷叶突变体sol1的表型分析与基因定位

适度卷曲有利于提高水稻叶片的光合效率,增加植株光合产物的有效积累量。我们利用甲基磺酸乙酯(EMS)处理籼型水稻保持系西农1B,获得一个稳定遗传的水稻半外卷叶突变体。该突变体从十叶期开始各叶片逐渐向外卷曲直至半卷状,并伴随茎秆半矮化和叶片披垂,暂被命名为semi-outcurved leaf 1(sol1)。与野生型(WT)相比,sol1的叶片卷曲指数均达到30%以上(P<0.01);倒一、倒二、倒三、倒四节节间长度和穗长极显著缩短,倒一、倒二、倒三叶的叶夹角显著或极显著增加;有效穗数、千粒重、每穗实粒数、结实率显著或极显著下降,一次枝梗数则增加11.3%(P<0.05)。sol1的蒸腾速率、胞间CO2浓度、气孔导度显著高于野生型。石蜡切片显示,sol1倒一叶的泡状细胞体积变小,数量显著增多,表皮细胞体积略微增大。遗传分析表明,sol1的半外卷叶性状受1对隐性核基因调控,定位于6号染色体标记JY6-3和JY6-10之间165 kb的物理范围内,共含15个注释基因。qRT-PCR结果表明,与泡状细胞相关的内卷基因和外卷叶基因RL14、Roc5、REL1在突变体sol1中呈不同程度的上调,NRL、BRD1、OsHox32、ADL1、LC2则呈不同程度的下调。研究结果为SOL1基因的克隆和功能研究奠定了基础。     

‘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。     

体细胞克隆荷斯坦奶牛出生后死亡犊牛内脏器官的组织病理学观察

为探究体细胞克隆荷斯坦奶牛出生后死亡的原因,对新生后死亡的克隆荷斯坦公牛和自然繁殖的荷斯坦公犊的主要组织器官进行比较。通过解剖和石蜡切片-HE(hematoxylin-eosin staining)染色技术,对主要的组织器官结构进行观察和分析。结果表明,新生后死亡的克隆荷斯坦公牛肺部结构清晰;肝脏的肝细胞明显肿大,出现脂肪轻度变性;肾小管上皮细胞出现变性;心肌的肌纤维间空隙增大;骨骼肌纤维间隙明显,空泡变性,这可能导致该克隆牛犊肌肉无力并功能不全;淋巴结皮髓质界限不分明,淋巴小结细胞较稀疏,生发中心不明显,淋巴窦细胞较少;脾脏内红细胞较少,这说明该克隆牛的造血功能可能不完善;胸腺的皮质部分和髓质部分界限不明显,嗜酸性胸腺小体不易辨认,可能发育不全。该克隆公牛免疫器官出现不同程度的发育不全现象较严重,这有可能是其出生后死亡率高的主要原因。     

miR-140-3p inhibits viability,invasion and migration of non-small-cell lung cancer A549 cells by targeting PD-L1

AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1.