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.     

ToxR负调控副溶血弧菌中c-di-GMP的合成

副溶血弧菌(Vibrio parahaemolyticus)是世界范围内引起海产品相关食物中毒的主要致病菌,具有很强的生物膜形成能力。ToxR是一种膜结合调控蛋白,对副溶血弧菌生物膜形成具有一定的调控作用,但具体机制尚未见报道。c-di-GMP是一种普遍存在于细菌中重要的第二信使,参与调控细菌的多种生物学行为包括生物膜的形成。本文探究ToxR对副溶血弧菌中c-di-GMP代谢的调控作用。利用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定副溶血弧菌野生株(wild-type,WT)和toxR突变株(ΔtoxR)中c-di-GMP水平的差异。挑选c-di-GMP代谢相关基因scrA、scrG和vpa0198为进一步研究的靶标,采用实时定量qPCR实验检测靶基因在WT和ΔtoxR中的转录水平差异;将靶基因调控区DNA序列克隆入pHRP309质粒中无启动子的β半乳糖苷酶基因上游,采用lacZ报告基因融合实验进一步研究ToxR对靶基因的转录调控关系;将重组质粒分别导入含有pBAD33或pBAD33-toxR的EC100lpir中,采用lacZ报告基因融合实验研究ToxR是否能在异体宿主中调控靶基因的表达;PCR扩增靶基因上游调控区DNA序列,并纯化His-ToxR蛋白,用凝胶阻滞实验(electrophoresis mobility shift assay,EMSA)研究His-ToxR与靶基因启动子区DNA序列是否具有结合作用。ELISA结果显示ΔtoxR中c-di-GMP含量显著性高于WT中的,说明ToxR抑制c-di-GMP的产生;实时定量qPCR结果表明WT中scrA、scrG和vpa0198的转录水平显著性高于ΔtoxR中的,表明ToxR抑制它们的转录;lacZ报告基因融合实验结果表明ToxR可抑制副溶血弧菌和EC100lpir中scrA、scrG和vpa0198的启动子区活性;EMSA实验显示His-ToxR能特异性地结合到scrA和scrG的上游调控区DNA序列上,而对vpa0198的上游调控区DNA序列无结合作用。综上所述,ToxR通过直接调控相关酶蛋白基因的转录来抑制副溶血弧菌内c-di-GMP的合成,从而有助于精确调控生物膜形成等细菌行为。     

Sleep Quality of Patients with Differentiated Thyroid Cancer

Objective

We aimed to measure prevalence of sleep disturbance in patients with differentiated thyroid cancer (DTC) by calculating Pittsburgh Sleep Quality Index (PSQI), and compare these data with patients with benign thyroid nodules or normal participants.

Methods

Three groups of patients participated in this cross-sectional study. In the first group, 162 patients with DTC received total thyroidectomy, and then ¹³¹I therapy. The second group consisted of 84 patients with benign thyroid nodules, who received partial thyroidectomy. The third group was 78 normal healthy control cases. PSQI was used to assess the sleep quality. Inter-group differences were analyzed by Kruskal-Wallis test or independent samples T test. χ² test was also used to check prevalence differences of poor sleep quality among the groups. Differences of PSQI score and poor sleep quality prevalence before and after ¹³¹I therapy in the same group of DTC participants were analyzed by paired T test and Mcnemar''s test.

Results

Higher PSQI score (7.59 ± 4.21) and higher rate of poor sleep quality (54.32%) were shown in DTC patients than in any other group. After ¹³¹I therapy, PSQI score and prevalence of poor sleep quality in DTC patients increased significantly to 8.78 ± 4.72 and 70.99%. Then DTC patients were divided into two subgroups based on their metastatic status. DTC patients with metastasis (87/162 cases, 53.70%) had significantly higher PSQI score (10.87 ± 5.18) and higher prevalence of poor sleep quality (79.31%).

Conclusion

DTC patients suffer from sleep disturbance, ¹³¹I therapy and awareness of metastatic status could worsen sleep problem. Psychological fear of cancer, nuclear medicine therapy and metastasis could be one major underlying reason. Longitude and interventional studies are necessary for further investigations.     

CalR激活副溶血弧菌Ⅵ型分泌系统1相关基因的转录

[目的]研究副溶血弧菌群体感应(quorum sensing,QS)系统核心调控子AphA和OpaR对calR基因以及CalR对Ⅵ型分泌系统l(type VI secretion system 1,T6SS1;vp1386-1420)相关基因的转录调控关系.[方法]提取副溶血弧菌野生株(wild-type,WT)和调控...     

High Expression of G6PD Increases Doxorubicin Resistance in Triple Negative Breast Cancer Cells by Maintaining GSH Level

Resistance to doxorubicin (DOX) remains a big challenge to breast cancer treatment especially for triple negative breast cancer (TNBC). Our previous study revealed that the antioxidant system plays an important role in conferring metastasis derived DOX resistance. In this study, we used two-dimensional difference gel electrophoresis (2D-DIGE) proteomics to compare the expression profiles of two generations of TNBC cell lines which have increased metastatic ability in nude mice and exhibited resistance to DOX. Through careful analyses, one antioxidant protein: glucose-6-phosphate dehydrogenase (G6PD) was identified with 3.2-fold higher level in metastatic/DOX-resistant 231-M1 than its parental 231-C3 cells. Analyses of clinical data showed that TNBC patients with higher G6PD levels exhibited lower overall survival than patients with lower G6PD level. Reducing G6PD expression by siRNA or inhibiting its activity with dehydroepiandrosterone (DHEA) significantly increased DOX''s cytotoxicity in both cell lines. Importantly, inhibiting G6PD''s activity with DHEA dramatically increased the apoptotic rate of 1.25 µM DOX from 2% to 54%. Our results suggest that high level of G6PD can help TNBC to resist DOX-induced oxidative stress. Thus, inhibiting G6PD shall be a good strategy to treat DOX-resistant TNBC.     

QsvR对副溶血弧菌VI型分泌系统1相关基因的转录调控

【目的】研究调控蛋白QsvR对副溶血弧菌VI型分泌系统1 （type VI secretion system 1,T6SS1）相关基因的转录调控关系。【方法】提取野生株（wild type,WT）和qsvR突变株（ΔqsvR）的总RNA,采用实时定量PCR （quantitative real-time PCR,qPCR）研究QsvR对靶基因的调控关系;进而采用引物延伸法定位靶基因的转录起始位点和核心启动子区,并根据引物延伸产物丰度判断QsvR对靶基因的调控关系;将靶基因的调控区DNA序列克隆入pHRP309质粒中的β-半乳糖苷酶基因上游（LacZ重组质粒）,并将重组质粒转化入WT和ΔqsvR中,通过LacZ报告基因融合试验研究QsvR对靶基因的调控关系;将LacZ重组质粒分别转化入含有pBAD33或pBAD33-qsvR的大肠杆菌100lpir中,进一步采用LacZ报告基因融合试验研究在异体宿主中QsvR对靶基因的调控关系;PCR扩增靶基因调控区DNA序列,同时表达并纯化His-QsvR重组蛋白,采用凝胶阻滞试验（electrophoresis mobility shift assay,EMSA）研究His-QsvR对靶基因调控区DNA序列是否具有直接的结合作用。【结果】qPCR结果显示,与WT相比,ΔqsvR中T6SS1相关基因VP1388 （操纵子VP1388-1390首基因）和hcp1 （操纵子VP1393-1406首基因）的转录水平显著性升高,表明QsvR抑制VP1388和hcp1的转录;引物延伸结果显示VP1388和hcp1各有一个转录起始位点,分别为C （-64）和T （-62）,且它们的转录活性受QsvR的抑制;LacZ报告基因融合试验结果显示QsvR可以抑制副溶血弧菌和EC100lpir中VP1388和hcp1的启动子区转录活性;EMSA结果显示His-QsvR对VP1388和hcp1的启动子区DNA序列具有直接的结合活性。【结论】QsvR对T6SS1相关操纵子VP1388-1390和VP1393-1406的转录具有直接的抑制作用。     

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.     

生物膜形成中间状态下副溶血弧菌的基因转录谱分析

【目的】探究生物膜形成中间状态下副溶血弧菌的差异基因表达情况,为今后研究生物膜形成调控机制提供基因信息。【方法】以非生物膜形成条件下为参照,采用Illumina HiSeq测序平台进行转录组测序（RNA sequencing,RNA-seq）研究,分析生物膜形成中间状态下副溶血弧菌的基因表达情况,并采用实时定量PCR （quantitative real-time PCR,qPCR）进行验证。【结果】本研究共获得979个差异显著性表达基因（differentially expressed gene,DEG）,其中下调基因379个,上调基因600个。基因本体（gene ontology,GO）分类分析结果显示,共有363个DEGs注释到分子功能、细胞组分和生物学过程3个一级分类和30个二级分类;京都基因和基因组百科全书（Kyoto encyclopedia of genes and genomes,KEGG）代谢途径分析结果显示,共有706个DEGs归到37个代谢通路中（Q value<0.05）,差异表达基因主要集中在细胞代谢和转运通路上;蛋白相邻类的聚簇（clusters of orthologous groups,COG）分类结果显示,有888个DEGs可归为20个类别,涉及DEGs最多的为氨基酸转运与代谢、一般功能预测基因、能量产生与转换以及未知功能基因。qPCR验证挑选的DEGs变化趋势均与RNA-seq的结果一致。此外,从转录组数据中共筛选出10个c-di-GMP代谢相关基因、1个侧生鞭毛蛋白基因（lafA）、13个极生鞭毛合成相关基因、6个荚膜多糖合成相关基因、6个胞外多糖合成相关基因、5个IV型菌毛合成相关基因、膜融合蛋白（membrane fusion protein,Mfp）基因（cpsQ-mfpABC）、14个III型分泌系统1（T3SS1）相关基因、14个Vp-PAI基因（1个tdh2和13个T3SS2基因）、3个VI型分泌系统1（T6SS1）相关基因、6个T6SS2基因、45个推定调控子基因和15个推定的外膜蛋白基因。【结论】生物膜形成引起副溶血弧菌基因表达谱发生明显变化,差异表达基因中包含生物膜形成相关基因、关键毒力基因和许多推定调控子基因等,这为后续研究生物膜形成调控机制提供重要信息。     

Transmembrane protein 106C promotes the development of hepatocellular carcinoma

Several protein-coding genes have been identified to play essential roles in cancer biology, and they are dysregulated in many tumors. Transmembrane protein 106C (TMEM106C) is differentially expressed in several human and porcine diseases; however, the expression and biological functions of TMEM106C in hepatocellular carcinoma (HCC) are not clear. In our study, we obtained paired tissue samples from patients undergoing resection for HCC and public databases, which were analyzed for TMEM106C expression using quantitative real-time polymerase chain reaction (qRT-PCR). We further conducted in vitro and in vivo experiments in HCC cell lines and nude mice, respectively, in which TMEM106C was overexpressed or knocked down. Cell-Counting Kit-8 and colony formation experiments were used to determine the influence of TMEM106C on cell proliferation, flow cytometric assays were used to detect the influence on cell cycle distribution and apoptosis, and transwell assays were used for detecting changes in cell migration and invasion. TMEM106C levels were significantly elevated in HCC tissues and cell lines from public databases and our collected specimens from patients. Moreover, higher TMEM106C expression levels predicted a poor prognosis in HCC patients in survival analysis. Overexpression of TMEM106C in HCC cells accelerated cell growth, migration, and invasion, but it inhibited cell apoptosis by targeting forkhead box O-1 (FOXO1) and FOXO3. Conversely, TMEM106C knockdown impeded cell proliferation and metastasis, whereas it enhanced the rate of apoptosis. More important, knockdown of the expression of TMEM106C in HCC cells inhibited the growth of xenograft tumors in vivo. Collectively, these results suggest that TMEM106C acts as an oncogene and can serve as a potential therapeutic target for HCC in the future.