Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown

Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78(DRi P78) and Na+-H+ exchanger regulatory factor 1(NHERF1) are the CXCR4 and CCR5 homo- or hetero-dimerinteracting proteins. DRi P78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRi P78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs(sh RNAs) targeting the DRi P78 and NHERF1, respectively, and constructed the p Lenti6/BLOCK-i T-DEST lentiviral plasmids expressing DRi P78 or NHERF1 sh RNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST(3). Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST(3) with DRi P78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.     

池塘混养模式下生态基对鲤生长及水质的影响

为探索池塘混养模式下生态基对鱼类生长的影响, 在6个土池中进行了一个饲养试验, 将土池分为2组, 一组为不放生态基的对照组, 另一组为放置生态基的生态基组, 每组3个重复。将尾均重为(310±11) g鲤3867尾、尾均重(810±15) g鲢及鳙370尾平均分别分为2组, 平均放养于6个土池中。对池塘鲤每天饲喂颗粒饲料3次, 饲养周期62d。在饲养期内, 每隔10天左右采集水样与底泥样品, 检测其中的浮游生物与微生物群落。在饲养结束后, 将试验鱼捕出并称重, 计算鲤的饲料效率。结果表明, 生态基组鲤鱼的增重率与饲料效率显著高于对照组; 生态基组的鲢、鳙末重显著低于对照组; 生态基组水体透明度与微生物群落多样性显著高于对照组; 生态基组浮游生物浓度低于对照组。结果表明, 在混养模式下土池中生态基的应用有利用促进鲤的生长, 然而池塘生态基的应用对于滤食性鲢、鳙的生长并无促进作用。     

黄鳝二价体上随机引物原位DNA合成的探讨

采用随机引物,在黄鳝二价体上建立了氚标记引物原位DNA合成技术,并详细探索了该技术的实验条件。其结果显示黄鳝二价体上DNA延伸效果十分明显,银颗粒基本上是非随机性的相间排列,而且主要特征是基本稳定的,据此可以进行染色体识别。此外,还讨论了建立该技术的重要意义。 Abstract:We have developed a technique of random-primed tritium labeled DNA extension on the pachytene bivalents of rice-field eels.The bivalents were not homogeneously labeled with this technique and the extension signals were very apparent and relatively stable,and these can be applied to identify the individual bivalent.The significance to develop this technique has also been discussed in detail.     

黄鳝二价染色体上地高辛配基标记随机引物原位DNA合成的研究

在黄鳝二价染色体上, 运用地高辛配基标记的随机引物原位DNA合成技术诱导出了明暗相间、基本稳定且类似于R带的带状结构。本文对该结果进行了较详细的分析和讨论。 Abstract:The dark bands alternating with light bands,relatively stable and R-band-like structure was showed on the pachytene bivalents of Rics-field eel(Monopterus albus Zuiew)by using the random-primed in situ digoxigenin-labelled DNA synthesis technique,and the results were analysed and discussed in detail.     

Revealing the Circadian Rhythm of Taliang Crocodile Newt(Liangshantriton taliangensis) During Breeding Migration Using a Novel Monitoring Technique

Many animals migrate during the breeding season. It is important to study the patterns of breeding migration of wild animals, especially rare and endangered species. However, current data acquisition methods are too coarse and imprecise for investigating the circadian rhythms of migrations in amphibians. Based on the frustrated total reflection image(FTRI), we developed a new device and recorded the precise migration time of an endangered salamander, Liangshantriton taliangensis.During the breed...     

应用BIGIS-4测序系统完成Glaciecola mesophilasp．nov．的全基因组测序和组装

BIGIS．4是中国新一代基于焦磷酸测序技术的测序仪．本实验利用BIGIS．4完成了对Glacielola mesophila sp．nov．（Gmn）的全基因组测序．Gmn是一株从海洋无脊椎动物体内分离到的革兰氏阴性菌．BIGIS-4测序得到152043个高质量的测序片段,平均读长406bp．测序片段由BIGIS-4系统后处理模块组装．除单核苷酸同聚体引起的测序错误外,测序结果中没有检测到其他低质量错误．Gmn基因组全长5144318bp,共注释得到4303个基因,其中有大量的代谢基因,与菌种在海洋表面非脊椎动物的生长环境相关．Gmn的冷适应和信号转导相关基因为其对海洋低温环境的适应提供了依据．     

miRNA-711-SP1-collagen-I pathway is involved in the anti-fibrotic effect of pioglitazone in myocardial infarction

Although microRNAs(miRNAs) have been intensively studied in cardiac fibrosis,their roles in drug-mediated anti-fibrotic therapy are still unknown.Previously,Pioglitazone attenuated cardiac fibrosis and increased miR-711 experimentally.We aimed to explore the role and mechanism of miR-711 in pioglitazone-treated myocardial infarction in rats.Our results showed that pioglitazone significantly reduced collagen-I levels and increased miR-711 expression in myocardial infarction heart.Pioglitazone increased the expression of miR-711 in cardiac fibroblasts,and overexpression of miR-711 suppressed collagen-I levels in angiotensin II(Ang II)-treated or untreated cells.Transfection with antagomir-711 correspondingly abolished the pioglitazone-induced reduction in collagen-I levels.Bioinformatics analysis identified SP1,which directly promotes collagen-I synthesis,as the putative target of miR-711.This was confirmed by luciferase assay and western blot analysis.Additionally,increased SP1 expression was attenuated by pioglitazone in myocardial infarction heart.Furthermore,transfection of antagomir-711 attenuated pioglitazone-reduced SP1 expression in cardiac fibroblasts with or without Ang II stimulation.We conclude that pioglitazone up-regulated miR-711 to reduce collagen-I levels in rats with myocardial infarction.The miR-711-SP1-collagen-I pathway may be involved in the anti-fibrotic effects of pioglitazone.Our findings may provide new strategies for miRNA-based anti-fibrotic drug research.     

Effect of vitrification on meiotic maturation and expression of genes in immature goat cumulus oocyte complexes

The aim of the study was to evaluate meiotic maturation, and expression of genes coding for oocyte secreted factors (GDF9, BMP15, TGFBR1, and BPR2) and apoptosis (BCL2, BAX and P53) after vitrification of immature goat cumulus oocyte complexes (COCs) and in vitro maturation. COCs were vitrified in a solution containing ethylene glycol, dimethyl sulfoxide and sucrose using either a conventional straw (CS), open pulled straw (OPS), cryoloop (CL), hemistraw (HS) or cryotop (CT). Freshly collected COCs (Control), COCs exposed to vitrification and dilution solutions without cryopreservation (EC) and vitrified-warmed COCs were matured in vitro for 27h. The viability of vitrified-warmed COCs 2 h post warming and in vitro maturation was similar for CL, HS and CT. The proportion of oocytes that extruded a 1st polar body and reached TI/MII was significantly higher with CT and HS followed by CL, OPS and CS. Gene expression of GDF9, BMP15, BMPR2, BAX and P53 were comparable to control levels for OPS, CL, HS and CT. The gene expression pattern in CS vitrified COCs was by contrast changed in that GDF9, BMP15, TGFBR1 and BAX were up regulated and BMPR2, BCL2 and P53 down regulated. In conclusion immature goat COCs vitrified using CT and HS showed that viability, maturation rates and expression of genes coding for oocyte secreted factors and apoptosis were similar to non-vitrified controls.