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.     

Clinical Features and Treatment of 2019-nCov Pneumonia Patients in Wuhan: Report of A Couple Cases

正Dear Editor,Till January 20, 2020, the 2019-new coronavirus(2019-nCoV) has caused more than one hundred cases in Wuhan(WMHC 2020). During a retrospective study of recent pneumonia patients in our department, we found two patients who are likely being infected with the 2019-nCoV.     

Evidence of compliance with and effectiveness of guidelines for noninvasive prenatal testing in China: a retrospective study of 189,809 cases

In China,the medical guidelines recommend performing noninvasive prenatal testing (NIPT) with caution for pregnant women aged 35 years or older.However,the Mother and Child Health Care Law suggests that all primiparous women whose age is older than 35 years undergo prenatal diagnosis.These two inconsistent suggestions/recommendations have made obstetricians confused about whether to offer NIPT to these older pregnant women.To face this issue and find out the solution we performed a retrospective study of 189,809 NIPT samples collected from 28 provincial-leveled administrative units in China.Of 1,564women with high-risk pregnancies who underwent NIPT,459 (29.3%) did not participate in follow-up.The compound sensitivity and specificity of NIPT for trisomies 21,18 and 13 detection was 99.1%(95%CI,98.0%-99.6%) and 99.9%(95%CI,98.8%-99.9%),respectively.In secundiparous women,NIPT showed high sensitivity and specificity similar to that in primiparous women.The observed risk for trisomies 21 and 18 significantly increased when the maternal age was 39 and older.After the publication of the current NIPT policy,the follow-up rate at our center was 97.9%;however,a large number of women are not in maternal and infant care networks nationwide,and that makes the follow-up rate outside our center relatively low.Our study shows that to balance the prevention of major aneuploidies and the limited resources for prenatal diagnosis,the cut-off age of 35for invasive prenatal diagnosis might be unnecessary.Although the NIPT guidelines are well written,how to practice it effectively,especially in less industrialized areas,is worth discussing.     

应用iTRAQ技术分析滩羊二毛期不同花穗型裘皮差异蛋白

为了解二毛期时滩羊串子花型裘皮与其它类型裘皮中蛋白质的差异,本试验采用iTRAQ技术及LC-MS/MS蛋白质组学研究方法,对串子花型、软大花型、绿豆丝型及其它不规则型花穗裘皮蛋白质进行鉴定和筛选,并运用Proteome Discoverer l.4软件进行定量分析,结合数据库搜索,鉴定出具有显著表达差异的蛋白,同时应用生物学技术对其进行GO和Pathway分析。结果显示4类花穗型裘皮共检测出2 886个蛋白,其中有135个、142个、113个差异蛋白分别存在于软大花型与串子花型、绿豆丝型与串子花型、其它不规则型与串子花型3个对比组中。对有表达差异的蛋白进行分析,发现膜联蛋白与血管内皮生长因子可能与软大花型毛股形成相关,KAP3和KAP6可能与滩羊串子花型毛股结构相关。研究发现滩羊不同二毛裘皮蛋白水平上的差异,可为选育优良的滩羊串子花型二毛裘皮提供理论基础。     

The use of SARS-CoV-2-related coronaviruses from bats and pangolins to polarize mutations in SARS-Cov-2

正Dear Editor,The coronavirus disease 2019 (COVID-19) caused by the SARS-CoV-2 coronavirus has become a global pandemic.The SARS-CoV-2 genome has a similarity of 96.2%to that of RaTG13, a bat SARS-CoV-2-related coronavirus detected in Rhinolophus affinis (Paraskevis et al., 2020; Zhou et al.,2020). The SARS-CoV-2 genome also has 85.5%-92.4%     

Depth‐dependent soil organic carbon dynamics of croplands across the Chengdu Plain of China from the 1980s to the 2010s

Agricultural soils have tremendous potential to sequester soil organic carbon (SOC) and mitigate global climate change. However, agricultural land use has a profound impact on SOC dynamics, and few studies have explored how agricultural land use combined with soil conditions affect SOC changes throughout the soil profile. Based on a paired soil resampling campaign in the 1980s and 2010s, this study investigated the SOC changes of the soil profile caused by agricultural land use and the correlations with parent material and topography across the Chengdu Plain of China. The results showed that the SOC content increased by 3.78 g C/kg in the topsoil (0–20 cm), but decreased in the 20–40 cm and 40–60 cm soil layers by 0.90 and 1.26 g C/kg respectively. SOC increases in topsoil were observed for all types of agricultural land. Afforestation on former agricultural land also caused SOC decreases in the 20–60 cm soil layers, while SOC decreases only occurred in the 40–60 cm soil layer for agricultural land using a traditional crop rotation (i.e. traditional rice–wheat/rapeseed rotation) and with rice–vegetable rotations converted from the traditional rotations. For each agricultural land use, SOC decreases in deep soils only occurred in high relief areas and in soils formed from Q4 (Quaternary Holocene) grey‐brown alluvium and Q4 grey alluvium that had a relatively low soil bulk density and clay content. The results indicated that SOC change caused by agricultural land use was depth dependent and that the effects of agricultural land use on soil profile SOC dynamics varied with soil characteristics and topography. Subsoil SOC decreases were more likely to occur in high relief areas and in soils with low soil bulk density and low clay content.     

New bioluminescence pyrophosphate assay for high‐sensitivity detection of food‐borne pathogens

Traditional methods of identifying food‐borne pathogens are time consuming and laborious, so innovative methods for their rapid identification must be developed. Testing for bioluminescence pyrophosphate is a convenient and fast method of detecting pathogens without complex equipment. However, the sensitivity of the method is not as high as that of other methods, and it has a very high detection limit. In this study, the method was optimized to improve its sensitivity. The shortcomings of the method were first identified and corrected using dATPαS instead of dATP for the polymerase chain reaction (PCR), therefore reducing the background signal. Also, when the DNA template extracted from the food‐borne pathogens was purified, the new bioluminescence pyrophosphate assay had a limit of detection of <10 copy/μl or 10 colony‐forming units/ml, and its sensitivity was higher than that of fluorescent real‐time quantitative PCR. Moreover, a single copy of a food‐borne pathogen could be detected when a single DNA template was included in the PCR. Salmonella was detected in and isolated from 60 samples of broiler chicken, and the accuracy of the results was verified using a culture method (GB 4789.4–2010). These results showed that the new bioluminescence pyrophosphate assay has the advantages of an intuitive detection process, convenient operation, and rapid measurements. Therefore, it can be used for the rapid detection of pathogenic bacteria and probiotics in various fields.