CO2对转基因鱼腥藻生长的影响

为了进一步探索转基因鱼腥藻高密度培养的方法,在小型气升式反应器中研究了CO2对转基因鱼腥藻7120培养的影响。结果发现转基因鱼腥藻培养过程中通入5％ CO2能促进藻细胞生长,12d生物量提高7．44％,由于光照限制,不能大幅提高15d收获生物量,但生长周期能缩短近20％;而高浓度(10％)的CO2抑制转基因鱼腥藻的生长。CO2是通过调节pH值和影响碳源利用来影响藻细胞生长的,合适浓度的CO2有利于转基因鱼腥藻的培养。     

环境因子对转基因鱼腥藻培养的影响

摇瓶中对转TNF-α基因鱼腥藻7120（Anabaenasp.PCC7120,pDC-TNF)混合培养条件进行了研究,在含蔗糖9g/L,NaNO32.25g/L的BG-11培养基混合培养时,得到最适培养条件接种量5%,光照强度为1000Lux,光/暗周期（光照时间/黑暗时间）12h/12h,温度25-30℃,自然初始pH值,100mL摇瓶装液量40mL,转基因鱼腥藻15d生物量可达到3g/L以上,可溶性蛋白含量接近30%,TNF表达水平大于22%,与自养相比,生物量增加82.06%,表达水平提高38.79%,证明混合营养型培养是转rhTNF-α基因鱼腥藻7120实现高密度,高表达培养的途径。     

碳氮源对转基因鱼腥藻Anabaena sp.PCC7120培养的影响

对碳源、氮源种类和用量对转rhTNF-α基因鱼腥藻7120（Anabaena sp.PCC7120）培养的影响进行了研究,发现最适碳源为蔗糖,最适氮源为NaNO3,最佳用量分别为9g/L和2．25g/L,此时生物量远高于自养方式,达2．52g/L,比相同条件下在BG－11培养基培养高71．66％,ATNF－α表达量为16％－22％,生物活性为10^5U/mg。     

利用气升式内环流生物反应器培养青蒿毛状根生产青蒿素

利用自制的气升式内环流生物反应器进行青蒿（ArtemisiaannuaL．）毛状根多层培养生产青蒿素。毛状根培养物在培养过程中均匀分布在生物反应器的筛网间，或以不锈钢网为附着点向四周生长，在25℃和12h／d光照周期下，经20d分批培养获得生物量干重22．57g／L，青蒿素产量374．4mg／L，并对培养过程中蔗糖、磷酸盐、硝酸盐和氨盐消耗的动力学进行了分析。     

转基因鱼腥藻7120适宜生长条件的初步研究

以前的研究报导了将人肿瘤坏死因子基因(TNF-α)成功转入鱼腥藻7120中,这项研究在实验室小规模范围内探讨了其转TNF-α基因鱼腥藻7120的适宜生长条件。结果表明,转基因鱼腥藻7120最适生长温度在25～30℃,最适pH7.0～7.5。不同光照强度下的净光合放氧速率测定结果表明,转基因鱼腥藻7120与野生型具有一致的光饱和点和光补偿点,且适合在10～700μE.m-2.s-1下生长。外加有机碳源(4g/L葡萄糖)一定程度上可以增加转基因鱼腥藻7120生长速度。对不同藻龄转基因鱼腥藻7120中TNF-α的累积量的检测发现:生长8d左右时转基因鱼腥藻7120中TNF-α累积量达到最大值。这将为产业化生产、适时收获转基因蓝藻提供理论指导,同时讨论了葡萄糖对转基因鱼腥藻7120代谢的调节。     

利用气升式内环流生物反应器培养青蒿毛状根生产青蒿素

利用自制的气升式内环流生物反应器进行青蒿(Artemisia annua L.)毛状根多层培养生产青蒿素。毛状根培养物在培养过程中均匀分布在生物反应器的筛网间,或以不锈钢网为附着点向四周生长,在25 ℃和12 h/d光照周期下,经20 d分批培养获得生物量干重22.57 g/L,青蒿素产量374.4 mg/L,并对培养过程中蔗糖、磷酸盐、硝酸盐和氨盐消耗的动力学进行了分析。     

蓝细菌病毒A-4L在鱼腥藻(Anabaena variabilis)藻苔中形成同心圆噬斑的成因

摘要:【目的】分析蓝细菌病毒A-4L在鱼腥藻(Anabaena sp.) PCC 7120藻苔中形成同心圆噬斑的原因,阐明A-4L的一个重要生物学特征,为分离、鉴定和筛选新的水华蓝藻病毒提供借鉴。【方法】用初始滴度为2.8×1010PFU/mL的A-4L悬液感染鱼腥藻,在不同时间点收集裂解液绘制一步生长曲线,获得A-4L对鱼腥藻的潜伏期和释放量。将适量A-4L悬液感染不同培养时间的藻苔,逐日观察和记录藻苔病变情况。培养藻苔并接种适量A-4L悬液,分别置于完全持续光照(Light:Dark=24 h:0 h,L:D=24 h:0 h)条件;完全周期光照(L:D=14 h:10h)条件;或前3 d周期光照转后3 d持续光照的条件下,比较不同光照条件对同心圆噬斑形成的影响。然后挑取单个噬斑进行扩大培养,纯化后,负染电镜观察A-4L的超微形态。【结果】A-4L的潜伏期为0.5－2h,释放量约为247 IU/cell (Infectious Units)。在周期光照条件下,藻苔接种A-4L 3－4 d后,出现同心圆噬斑,且同心圆数量与攻毒后的天数(n)有相关性,为“n－1”;同心圆间距约为3 mm。与周期光照条件相比,在持续光照条件下未形成同心圆噬斑。而在周期光照条件转持续光照条件下,由先周期光照时所形成的同心圆在转持续光照后逐渐消失,证实同心圆噬斑的形成依赖于周期光照。负染电镜观察显示A-4L具有一个近似球形的头部,直径约为50 nm以及长度约为10 nm的尾部,形态与短尾蓝细菌病毒相似。【结论】A-4L是一株能形成同心圆噬斑的蓝细菌病毒,并揭示其同心圆噬斑形成的关键条件是周期光照。     

布朗葡萄藻的培养基选择及其产物代谢规律

通过对布朗葡萄藻分别在Chu13、Chu13×2和BG-11培养基中培养结果的比较,发现在气升式光照生物反应器中Chu13培养基最有利于布朗葡萄藻的生长和烃的合成,培养15d后,其生物量和粗烃质量分数分别为1.82g/L和58.7%;棕榈酸、油酸和亚麻酸是布朗葡萄藻的主要脂肪酸组成,Chu13培养获得的藻体不饱和脂肪酸比例最高。Chu13培养基中布朗葡萄藻代谢规律的研究表明:粗烃含量随着生物量的增加而逐渐增大,15d后粗烃产量达到最大值1.07g/L,不同生长周期烃的组成保持一致,布朗葡萄藻的烃主要由C33H56和C34H58组成;在布朗葡萄藻生长周期中,不饱和脂肪酸的比例显著上升,培养15d达到64%以上。     

蓝细菌病毒A-4L在鱼腥藻(Anabaena variabilis)藻苔中形成同心圆噬斑的成因

【目的】分析蓝细菌病毒A-4L在鱼腥藻(Anabaena sp.)PCC 7120藻苔中形成同心圆噬斑的原因,阐明A-4L的一个重要生物学特征,为分离、鉴定和筛选新的水华蓝藻病毒提供借鉴。【方法】用初始滴度为2.8×1010PFU/mL的A-4L悬液感染鱼腥藻,在不同时间点收集裂解液绘制一步生长曲线,获得A-4L对鱼腥藻的潜伏期和释放量。将适量A-4L悬液感染不同培养时间的藻苔,逐日观察和记录藻苔病变情况。培养藻苔并接种适量A-4L悬液,分别置于完全持续光照(Light∶Dark=24 h∶0 h,L∶D=24 h∶0 h)条件;完全周期光照(L∶D=14 h∶10 h)条件;或前3 d周期光照转后3 d持续光照的条件下,比较不同光照条件对同心圆噬斑形成的影响。然后挑取单个噬斑进行扩大培养,纯化后,负染电镜观察A-4L的超微形态。【结果】A-4L的潜伏期为0.5-2 h,释放量约为247 IU/cell(Infectious Units)。在周期光照条件下,藻苔接种A-4L 3-4 d后,出现同心圆噬斑,且同心圆数量与攻毒后的天数(n)有相关性,为"n-1";同心圆间距约为3 mm。与周期光照条件相比,在持续光照条件下未形成同心圆噬斑。而在周期光照条件转持续光照条件下,由先周期光照时所形成的同心圆在转持续光照后逐渐消失,证实同心圆噬斑的形成依赖于周期光照。负染电镜观察显示A-4L具有一个近似球形的头部,直径约为50 nm以及长度约为10 nm的尾部,形态与短尾蓝细菌病毒相似。【结论】A-4L是一株能形成同心圆噬斑的蓝细菌病毒,并揭示其同心圆噬斑形成的关键条件是周期光照。     

两种微藻在光生物反应器中的生长和胞内多糖含量研究

利用气升式光生物反应器就光照周期、光质和通气速度对湛江叉鞭金藻和盐藻的生长和胞内多糖含量进行了研究。结果表明：(1)湛江叉鞭金藻和盐藻在亮暗比分别为12L：12D和18L：6D时生长最快,在18L：6D下两种微藻多糖含量较高;(2)不同光质比较,叉鞭金藻在蓝光、红光下,盐藻在红光、白光下有较高的生长速率和多糖含量;(3)600ml／min的通气速度可以获得较好的培养效果。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.