Long Term Culture of Human Kidney Proximal Tubule Epithelial Cells Maintains Lineage Functions and Serves as an Ex vivo Model for Coronavirus Associated Kidney Injury

The mechanism of how SARS-CoV-2 causes severe multi-organ failure is largely unknown. Acute kidney injury(AKI) is one of the frequent organ damage in severe COVID-19 patients. Previous studies have shown that human renal tubule cells could be the potential host cells targeted by SARS-CoV-2. Traditional cancer cell lines or immortalized cell lines are genetically and phenotypically different from host cells. Animal models are widely used, but often fail to reflect a physiological and pathogenic status because of species tropisms. There is an unmet need for normal human epithelial cells for disease modeling. In this study, we successfully established long term cultures of normal human kidney proximal tubule epithelial cells(KPTECs) in 2 D and 3 D culture systems using conditional reprogramming(CR) and organoids techniques.These cells had the ability to differentiate and repair DNA damage, and showed no transforming property. Importantly, the CR KPTECs maintained lineage function with expression of specific transporters(SLC34 A3 and cubilin). They also expressed angiotensin-converting enzyme 2(ACE2), a receptor for SARS-CoV and SARS-CoV-2. In contrast, cancer cell line did not express endogenous SLC34 A3, cubilin and ACE2. Very interestingly, ACE2 expression was around twofold higher in 3 D organoids culture compared to that in 2 D CR culture condition. Pseudovirion assays demonstrated that SARS-CoV spike(S) protein was able to enter CR cells with luciferase reporter. This integrated 2 D CR and 3 D organoid cultures provide a physiological ex vivo model to study kidney functions, innate immune response of kidney cells to viruses, and a novel platform for drug discovery and safety evaluation.     

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...     

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

Virologica Sinica - The corrected legend is given below.     

Epidemiological,Clinical and Serological Characteristics of Children with Coronavirus Disease 2019 in Wuhan: A Single-centered,Retrospective Study

In December 2019, SARS-CoV-2 was first detected in the samples obtained from three adult patients who suffered from an unknown viral pneumonia in Wuhan (Li et al. 2020). This unknown viral pneumonia is further named as coronavirus disease 2019 (COVID-19) by the World Health Organization. To date, the number of new COVID-19 cases has continued to skyrocket and the impact of SARS-CoV-2 on humans is far greater than any pathogen of this century in both breadth and depth. Previous studies have shown that adults with COVID-19 have symptoms of fever, dry cough, dyspnea, fatigue and lymphocytopenia. Moreover, COVID-19 is more likely to cause death in the elderly, especially those with chronic comorbidities (Huang et al. 2020). In Wuhan, more than 50, 000 COVID-19 cases have been confirmed, including over 780 pediatric patients, and only one child death case (Lu et al. 2020). Although the number of children cases was far fewer than that of adults, COVID-19 might endanger children's health and the information on children remains limited, especially in serological study. In the retrospective study, the investigators analyzed the epidemiological, clinical and serological characteristics of children with COVID-19 in Wuhan in the early stages of the outbreak, which might provide theoretical and practical help in controlling COVID-19 and similar emerging infectious diseases in the future.     

Epidermal growth factor improves intestinal morphology by stimulating proliferation and differentiation of enterocytes and mTOR signaling pathway in weaning piglets

Epidermal growth factor(EGF) has been shown to improve piglet intestinal morphology and epithelial recovery. In an attempt to further understand the mechanisms behind these improvements, this study tested the hypothesis that dietary EGF may affect intestinal morphology by stimulating the proliferation and differentiation of enterocytes in weaning piglets. In piglets receiving200 μg kg–1 EGF, crypt depth and villus height increased(P0.05). Adding 400 μg kg–1 EGF increased villus height-to-crypt depth ratio(P0.05), but reduced crypt depth(P0.05). Dietary supplementation with 200 μg kg–1 EGF significantly increased the number of Ki67-positive cells(P0.01) and tended to increase the mRNA level of proliferating cell nuclear antigen(P0.10).However, this supplementation decreased the expression level of intestinal fatty acid-binding protein(P0.05). Piglets fed with400 μg kg–1 EGF had an increased mRNA level of intestinal alkaline phosphatase(P0.05). The phosphorylation of m TOR(mammalian target of rapamycin) was observed in the 200 μg kg–1 EGF group. These results suggest that dietary supplementation with a low level of EGF improved piglet intestinal morphology through stimulating the proliferation and differentiation of enterocytes, and the mTOR signaling pathway may partly be involved in this process.     

玉米种子丸粒化包衣处理对草地贪夜蛾的防治效果

本研究明确了乙基多杀菌素和氯虫苯甲酰胺丸粒化包衣后对玉米种子的安全性和玉米幼苗对草地贪夜蛾的防治效果,为玉米苗期草地贪夜蛾防治提供新技术。首先在室内验证了乙基多杀菌素和氯虫苯甲酰胺拌土后对草地贪夜蛾的防治效果,又进行了种子丸粒化包衣处理,测定了丸粒化包衣方式下药剂对种子萌发的影响和对草地贪夜蛾的防治效果。结果表明,250 mg/kg的乙基多杀菌素和氯虫苯甲酰胺拌土处理后,草地贪夜蛾对14天的玉米苗的取食率分别为1.67%、5.01%,与对照组取食率27.02%差异显著。药种比为1:100和1:200的乙基多杀菌素和氯虫苯甲酰胺丸粒化包衣处理组与对照组的发芽率之间无显著差异。乙基多杀菌素和氯虫苯甲酰胺丸粒化包衣处理组在播种后第7天对草地贪夜蛾广州种群防治效果显著,其中药种比为1:100的乙基多杀菌素丸粒化包衣处理组致死率最高为89%;两药剂丸粒化包衣处理组在播种后第21天对草地贪夜蛾云南种群仍有较好防治效果,两个浓度的乙基多杀菌素丸粒化包衣处理对草地贪夜蛾的致死率均达100%。本研究表明玉米种子丸粒化包衣技术可有效防控玉米苗期草地贪夜蛾的危害。     

Molecular phylogeny and species delimitation of Stachyuraceae: Advocating a herbarium specimen‐based phylogenomic approach in resolving species boundaries

Species concept and delimitation are fundamental to taxonomic and evolutionary studies. Both inadequate informative sites in the molecular data and limited taxon sampling have often led to poor phylogenetic resolution and incorrect species delineation. Recently, the whole chloroplast genome sequences from extensive herbarium specimen samples have been shown to be effective to amend the problem. Stachyuraceae are a small family consisting of only one genus Stachyurus of six to 16 species. However, species delimitation in Stachyurus has been highly controversial because of few and generally unstable morphological characters used for classification. In this study, we sampled 69 individuals of seven species (each with at least three individuals) covering the entire taxonomic diversity, geographic range, and morphological variation of Stachyurus from herbarium specimens for genome‐wide plastid gene sequencing to address species delineation in the genus. We obtained high‐quality DNAs from specimens using a recently developed DNA reconstruction technique. We first assembled four whole chloroplast genome sequences. Based on the chloroplast genome and one nuclear ribosomal DNA sequence of Stachyurus, we designed primers for multiplex polymerase chain reaction and high throughput sequencing of 44 plastid loci for species of Stachyurus. Data of these chloroplast DNA and nuclear ribosomal DNA internal transcribed spacer sequences were used for phylogenetic analyses. The phylogenetic results showed that the Japanese species Stachyurus praecox Siebold & Zucc. was sister to the rest in mainland China, which indicated a typical Sino‐Japanese distribution pattern. Based on diagnostic morphological characters, distinct distributional range, and monophyly of each clade, we redefined seven species for Stachyurus following an integrative species concept, and revised the taxonomy of the family based on previous reports and specimens, in particular the type specimens. Furthermore, our divergence time estimation results suggested that Stachyuraceae split from its sister group Crossosomataceae from the New World at ca. 54.29 Mya, but extant species of Stachyuraceae started their diversification only recently at ca. 6.85 Mya. Diversification time of Stachyurus in mainland China was estimated to be ca. 4.45 Mya. This research has provided an example of using the herbarium specimen‐based phylogenomic approach in resolving species boundaries in a taxonomically difficult genus.     

全反式维甲酸对骺软骨细胞生物学行为及其功能影响的研究

该研究主要探讨了体外高浓度全反式维甲酸(all-trans retinoic acid,ATRA)对SD大鼠骺软骨细胞生物学性状和功能的影响以及体内ATRA对SD大鼠胫骨生长板的影响。以SD大鼠骺软骨细胞为研究对象、ATRA为干预因素,采用CCK-8、细胞流式术、HE染色、Annexin V-FITC细胞凋亡流式检测术、Hoechst染色、细胞划痕、Transwell实验分别评估ATRA处理后细胞的增殖、周期、形态、凋亡及迁移情况,Western blot检测蛋白聚糖、Ⅱ型胶原、X型胶原等相关功能蛋白的变化;以3周雄性SD大鼠为实验对象,分为对照组、60 mg/kg·d ATRA组、80 mg/kg·d ATRA组,进行10天连续ATRA灌胃处理,测量每只SD大鼠灌胃第1天、第10天的头尾长,处理10天后对胫骨生长板进行HE染色。结果表明,ATRA作用SD大鼠骺软骨细胞后,增殖能力减弱且细胞周期被阻滞在S期(P<0.01),细胞形态由三角形、多边形变为长条状,凋亡的发生增多(P<0.01),迁移能力受到抑制(P<0.05)以及Western blot结果显示蛋白聚糖、Ⅱ型胶原、X型胶原等功能相关蛋白较对照组表达均明显降低(P<0.01);对SD大鼠进行ATRA灌胃处理后,与对照组比较,60 mg/kg·d ATRA组和80 mg/kg·d ATRA组的头尾长均变短(P<0.01);胫骨生长板HE染色显示,ATRA灌胃组的生长板变窄甚至闭合。该研究证实了体外高浓度ATRA能够对SD大鼠骺软骨细胞的增殖、迁移起抑制作用,同时能够诱导凋亡,降低相关功能蛋白的表达,在SD大鼠体内证实,过量ATRA可影响生长板软骨内成骨过程,最终使生长板部分或全部提前闭合,进而影响SD大鼠身长的增长。     

Alpine grassland plants grow earlier and faster but biomass remains unchanged over 35 years of climate change

Satellite data indicate significant advancement in alpine spring phenology over decades of climate warming, but corresponding field evidence is scarce. It is also unknown whether this advancement results from an earlier shift of phenological events, or enhancement of plant growth under unchanged phenological pattern. By analyzing a 35‐year dataset of seasonal biomass dynamics of a Tibetan alpine grassland, we show that climate change promoted both earlier phenology and faster growth, without changing annual biomass production. Biomass production increased in spring due to a warming‐induced earlier onset of plant growth, but decreased in autumn due mainly to increased water stress. Plants grew faster but the fast‐growing period shortened during the mid‐growing season. These findings provide the first in situ evidence of long‐term changes in growth patterns in alpine grassland plant communities, and suggest that earlier phenology and faster growth will jointly contribute to plant growth in a warming climate.