Maturation and fertilization of porcine oocytes in vitro

Four experiments were conducted to study 1) factors affecting porcine oocyte maturation in culture medium and 2) a new method for oocyte maturation outside the porcine body. In Experiment 1, five groups of oocytes were cultured in m-TCM199 or m-KRB medium for 24 to 28, 32 to 36 or 40 to 42 hours and then were fertilized in vitro. The cleavage rate (two to four-cell stage) of oocytes cultured for 32 to 36 hours was significantly higher than those of the other oocytes. The results indicate that a suitable culture period for the in vitro maturation of porcine oocytes is 32 to 36 hours. In Experiment 2, four groups of oocytes were cultured in m-KRB or m-KRB supplemented with PFF, PMSG or FSH for in vitro maturation, and the cleavage rates of oocytes were 7.94, 22.56, 30.23 and 23.26%, respectively, after in vitro fertilization. The results show that porcine follicular fluid (PFF) and gonadotrophins added to the culture medium promote porcine oocyte maturation in vitro. In Experiment 3, oocytes were cultured in m-KRB or m-TCM199, supplemented with both gonadotrophin and pocine folliclar fluid for maturation in vitro. After fertilization in vitro, the cleavage rates of oocytes were 26.32 and 27.93% for the two media. The results indicate that the difference between m-KRB and m-TCM199 was insignificant when the media were used to culture porcine oocytes. But there was a significant difference when PFF and gonadotrophins were added to the basic media. In Experiment 4, porcine oocytes were transferred into the reproductive tracts of other animals for maturation. After 34 to 36 hours, the oocytes were collected and fertilized in vitro. The cleavage rates of oocytes were 10.42, 28.45, 3.33 and 36.36%, respectively, for the oocytes matured in mouse uterine horns, rat uterine horns, rat oviducts or rabbit oviducts. The results show that porcine oocytes can be matured in the reproductive tracts of other animals.     

小黑杨花粉植株H_1株系间变异及其选择利用的研究

小黑杨(Populus xiaohei T.S.Hwang ex C.Wang et Tung)花药离体培养获得单倍体植株,经14年的生长观察,发现花粉植株间表现出明显分离。选出19个株系进行田间比较试验,统计分析,结果发现材积生长差异显著。可从中选出优良株系加以利用,并提出杨树单倍体育种程序。     

Luteolin and Apigenin Attenuate 4-Hydroxy-2-Nonenal-Mediated Cell Death through Modulation of UPR,Nrf2-ARE and MAPK Pathways in PC12 Cells

Luteolin and apigenin are dietary flavones and exhibit a broad spectrum of biological activities including antioxidant, anti-inflammatory, anti-cancer and neuroprotective effects. The lipid peroxidation product 4-hydroxy-2-nonenal (4-HNE) has been implicated as a causative agent in the development of neurodegenerative disorders. This study investigates the cytoprotective effects of luteolin and apigenin against 4-HNE-mediated cytotoxicity in neuronal-like catecholaminergic PC12 cells. Both flavones restored cell viability and repressed caspase-3 and PARP-1 activation in 4-HNE-treated cells. Luteolin also mitigated 4-HNE-mediated LC3 conversion and reactive oxygen species (ROS) production. Luteolin and apigenin up-regulated 4-HNE-mediated unfolded protein response (UPR), leading to an increase in endoplasmic reticulum chaperone GRP78 and decrease in the expression of UPR-targeted pro-apoptotic genes. They also induced the expression of Nrf2-targeted HO-1 and xCT in the absence of 4-HNE, but counteracted their expression in the presence of 4-HNE. Moreover, we found that JNK and p38 MAPK inhibitors significantly antagonized the increase in cell viability induced by luteolin and apigenin. Consistently, enhanced phosphorylation of JNK and p38 MAPK was observed in luteolin- and apigenin-treated cells. In conclusion, this result shows that luteolin and apigenin activate MAPK and Nrf2 signaling, which elicit adaptive cellular stress response pathways, restore 4-HNE-induced ER homeostasis and inhibit cytotoxicity. Luteolin exerts a stronger cytoprotective effect than apigenin possibly due to its higher MAPK, Nrf2 and UPR activation, and ROS scavenging activity.     

The synergy of germline C634Y and V292M RET mutations in a northern Chinese family with multiple endocrine neoplasia type 2A

Genetic analysis for germline mutations of RET proto‐oncogene has provided a basis for individual management of medullary thyroid carcinoma (MTC) and pheochromocytoma. Most of compound mutations have more aggressive phenotypes than single point mutations, but the compound C634Y/V292M variant in MTC has never been reported. Thus, we retrospectively investigated synergistic effect of C634Y and V292M RET germline mutations in family members with multiple endocrine neoplasia type 2A. Nine of 14 family members in a northern Chinese family underwent RET mutation screening using next‐generation sequencing and PCR followed by direct bidirectional DNA sequencing. Clinical features of nine individuals were retrospectively carefully reviewed. In vitro, the scratch‐wound assay was used to investigate the difference between the cells carrying different mutations. We find no patients died of MTC. All 3 carriers of the V292M variant were asymptomatic and did not have biochemical or structural evidence of disease (age: 82, 62 and 58). Among 4 C634Y mutation carriers, 2 patients had elevated calcitonin with the highest (156 pg/mL) in an 87‐year‐old male. Two carriers of compound C634Y/V292M trans variant had bilateral MTC with pheochromocytoma or lymph node metastasis (age: 54 and 41 years, respectively). Further, the compound C634Y/V292M variant had a faster migration rate than either single point mutation in vitro (P < .05). In conclusion, the V292M RET variant could be classified as ‘likely benign’ according to ACMG (2015). The compound variant V292M/C634Y was associated with both more aggressive clinical phenotype and faster cell growth in vitro than was either single mutation.     

MicroRNA‐29b‐3p suppresses oral squamous cell carcinoma cell migration and invasion via IL32/AKT signalling pathway

Oral squamous cell carcinoma (OSCC) is aggressive accompanied with poor prognosis. We previously isolated the most invasive cells resembling the invasive tumour front by microfluidic technology and explored their differentially expressed microRNAs (miRNAs) in our previous work. Here, we verified the miR‐29b‐3p as a guarder that suppressed migration and invasion of OSCC cells and was down‐regulated in the most invasive cells. Besides that, the invasion suppression role of miR‐29b‐3p was achieved through the IL32/AKT pathway. Thus, miR‐29b‐3p and IL32 might serve as therapeutic targets for blocking the progression and improving the outcome of OSCC.     

Establishment of Murine Infection Models with Biological Clones of Dengue Viruses Derived from a Single Clinical Viral Isolate

Virologica Sinica - Dengue virus (DENV) is a single-stranded RNA virus transmitted by mosquitoes in tropical and subtropical regions. It causes dengue fever, dengue hemorrhagic fever and dengue...     

High-dose zolpidem abuse in a patient with insomnia comorbid with major depressive disorder

Sleep and Biological Rhythms - A 39-year-old married woman was referred to a psychiatry outpatient clinic for treatment of major depressive disorder (MDD). The dose of zolpidem she had been taking...     

Increased high‐latitude photosynthetic carbon gain offset by respiration carbon loss during an anomalous warm winter to spring transition

Arctic and boreal ecosystems play an important role in the global carbon (C) budget, and whether they act as a future net C sink or source depends on climate and environmental change. Here, we used complementary in situ measurements, model simulations, and satellite observations to investigate the net carbon dioxide (CO₂) seasonal cycle and its climatic and environmental controls across Alaska and northwestern Canada during the anomalously warm winter to spring conditions of 2015 and 2016 (relative to 2010–2014). In the warm spring, we found that photosynthesis was enhanced more than respiration, leading to greater CO₂ uptake. However, photosynthetic enhancement from spring warming was partially offset by greater ecosystem respiration during the preceding anomalously warm winter, resulting in nearly neutral effects on the annual net CO₂ balance. Eddy covariance CO₂ flux measurements showed that air temperature has a primary influence on net CO₂ exchange in winter and spring, while soil moisture has a primary control on net CO₂ exchange in the fall. The net CO₂ exchange was generally more moisture limited in the boreal region than in the Arctic tundra. Our analysis indicates complex seasonal interactions of underlying C cycle processes in response to changing climate and hydrology that may not manifest in changes in net annual CO₂ exchange. Therefore, a better understanding of the seasonal response of C cycle processes may provide important insights for predicting future carbon–climate feedbacks and their consequences on atmospheric CO₂ dynamics in the northern high latitudes.     

Fluorescence characterization and microbial degradation of dissolved organic matter leached from salt marsh plants in the Yellow River Delta

盐沼植被是沿海水体中溶解有机物(Dissolved Organic Matter, DOM)的重要贡献者。然而,不同盐沼植物释放DOM的动力学特征尚缺乏系统研究和比较。黄河三角洲湿地是中国东海岸面积最大,保护最完善的沿海生态系统之一。本论文研究了2016年10月从黄河三角洲(Yellow River Delta, YRD)盐沼中采集的三种常见沼泽植物(芦苇(Phragmites australis),碱蓬(Suaeda salsa)和獐茅(Aeluropus littoralis)的DOM释放过程。通过测定溶解有机碳(Dissolved Organic Carbon, DOC)和溶解氮(Dissolved Nitrogen, DN)发现,植物叶片释放的DOM浓度远高于其根和茎。在27天的培养期内,平均有15%的生物碳和30%的生物氮以DOC和DN的形式通过植物叶片释放。从植物中释放的DOM非常不稳定,在27天的培养期内,细菌共消耗了92.4%–98.1%的DOC和88.0%–94.6%的DN。植物释放DOM的荧光特性表明,发色溶解的有机物(Chromophoric Dissolved Organic Matter, CDOM)是DOM的主要组分,而类蛋白组分是植物释放CDOM的主要组分。细菌的降解作用改变了DOM的荧光性质和化学组成。上述的室内研究结果得到了实地调查的充分支撑,表明在深秋时期黄河口湿地有大量DOM溢出。本研究结果表明,盐沼植物释放的DOM是沼泽和沿海水域DOC和DN的重要来源,而且易降解DOC和DN为黄河口湿地和邻近沿海水域中的微生物群落提供了重要的食物来源。     

黑斑侧褶蛙蝌蚪断尾后的补偿生长和发育研究

动物在经历不利的生长条件或环境后往往出现补偿生长。研究了黑斑侧褶蛙(Pelophylax nigromaculatus)蝌蚪经历来自食蚊鱼(Gambusia affinis)捕食造成断尾损伤后的补偿生长模式、断尾损失对蝌蚪游泳能力以及变态时间与大小的影响。结果表明,经历捕食压力后,全尾组和1/3断尾组蝌蚪的体长显著大于1/2断尾组蝌蚪的体长,全尾组和1/3断尾组之间的蝌蚪体长差异不显著;第19天时,1/2断尾组蝌蚪经过补偿生长后体长显著大于全尾组蝌蚪,1/3断尾组与全尾组和1/2断尾组之间的蝌蚪体长差异均不显著;三个实验组之间蝌蚪尾长差异不显著;全尾组蝌蚪的疾游速显著大于1/2断尾组蝌蚪的疾游速,1/3断尾组蝌蚪的疾游速与全尾组和1/2断尾组之间差异不显著,表明严重断尾对蝌蚪疾游速产生了消极影响。三个实验组蝌蚪的变态时间和变态前后形态差异均不显著。黑斑侧褶蛙蝌蚪能够在变态前调整生长轨迹补偿早期捕食风险造成的生长损失,断尾损失并不影响变态时间与大小,断尾超过一半的蝌蚪经过补偿生长后仍要付出一定的运动代价。