Detectable Serum Severe Acute Respiratory Syndrome Coronavirus 2 Viral Load (RNAemia) Is Closely Correlated With Drastically Elevated Interleukin 6 Level in Critically Ill Patients With Coronavirus Disease 2019.

BACKGROUND: Although the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral load in respiratory specimens has been widely used to diagnose coronavirus disease 2019 (COVID-19), it is undeniable that serum SARS-CoV-2 nucleic acid (RNAemia) could be detected in a fraction of COVID-19 patients. However, it is not clear whether testing for RNAemia is correlated with the occurrence of cytokine storms or with the specific class of patients. METHODS: This study enrolled 48 patients with COVID-19 admitted to the General Hospital of Central Theater Command, People's Liberation Army, a designated hospital in Wuhan, China. The patients were divided into 3 groups according to the Diagnosis and Treatment of New Coronavirus Pneumonia (sixth edition) guidelines issued by the National Health Commission of China. Clinical and laboratory data were collected, and the serum viral load and interleukin 6 (IL-6) level were determined. RESULTS: Analysis of clinical characteristics of 48 cases of COVID-19 showed that RNAemia was diagnosed only in the critically ill group and seemed to reflect the severity of the disease. Furthermore, the level of the inflammatory cytokine IL-6 in critically ill patients increased significantly, almost 10 times that in other patients. More importantly, the extremely high IL-6 level was closely correlated with the detection of RNAemia (R = 0.902). CONCLUSIONS: Detectable serum SARS-CoV-2 RNA (RNAemia) in patients with COVID-19 was associated with elevated IL-6 concentration and poor prognosis. Because elevated IL-6 may be part of a larger cytokine storm that could worsen outcome, IL-6 could be a potential therapeutic target for critically ill patients with an excessive inflammatory response.

Epidemiological and etiological characteristics of hand, foot, and mouth disease in Wuhan, China from 2012 to 2013: outbreaks of coxsackieviruses A10.

Hand-foot-mouth disease (HFMD) is a common infectious disease which often occurs in young children. It is caused by enteroviruses, most commonly enterovirus71 (EV71) and Coxsackievirus A16 (CVA16). The present study focuses on the molecular epidemiology of the pathogen of HFMD in the Wuhan region of China during the period 2012 to 2013. A total of 463 viruses were isolated from throat swab of 3,208 HFMD patients and analyzed by quantitative RT-PCR with all sets of specific primers for EV71, CVA16, and pan-enterovirus. Of the 463 viruses, 111 (21.2%) were EV71, 52 (9.6%) were CVA16, and 300 (69.2%) were pan-enterovirus. In pan-enterovirus isolations 190 (52.8%) were CVA10, 50 (13.9%) were CVA4, 30 were CB2, 17 were CB3, 13 were CB5 identified by VP4 gene sequencing. Eleven EV71 isolates were complete genome sequenced and phylogenetic analysis revealed that the EV71 strains that circulated in Wuhan belonged to the C4 subgenotype. Among the 190 CVA10 isolations, 187 CVA10 strains have the same nucleotide sequence, the other three CVA10 strains belongs to another type of nucleotide sequence. Phylogenetic analysis based on 19 CVA10 isolations suggested that they belonged to the clade of Chinese strains, but form different clusters isolated from Japan, Europe. This study showed that EVA71 and CVA16 were detected as the predominant viruses (>60%) in 2012 and the total reported HFMD cases attained a peak in June and July. In contrast, CVA10 was also detected during April 2012 and replaced EVA71 and CVA16 as the major HFMD-associated pathogen from May 2013.

Multiomics analysis of metabolic heterogeneity in cervical cancer cell lines with or without HPV.

Metabolomics analysis revealed the metabolic heterogeneity of cervical cancer (CC) cell lines C33A and CaSki, and their molecular mechanisms were explored. Using the modified Bligh-Dyer method, the endogenous metabolites of C33A and CaSki cells were divided into polar and nonpolar fractions. The metabolites were analysed by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Then, the differential metabolites were screened by combining multivariate statistical analysis and volcano maps, and functional enrichment and pathway analysis of the differential metabolites were performed. Finally, association analysis was carried out in combination with transcriptomics, and the important differential metabolisms were experimentally verified by real-time PCR (RT-qPCR) and oil red staining. The results showed that between the C33A and CaSki cell lines, there were significant differences in amino acids, nucleotides and lipids, such as in threonine, arachidonic acid and hypoxanthine, in the metabolic pathways. These compounds could be used as markers of differences in cellular metabolism. The heterogeneity of lipid metabolism accounted for 87.8%, among which C33A cells exhibited higher contents of fatty acid polar derivatives, while CaSki cells showed higher contents of free fatty acids and glycerides. Based on correlation analysis of the above metabolic differences in HPV pathways as well as lipid metabolism-related genes, p53 and the genes involved in lipid metabolism pathways, such as Peroxisome Proliferator Activated Receptor Gamma(PPARG) and stearoyl-CoA desaturase (SCD), are relevant to the metabolic heterogeneity of the cells. The differential expression of some genes was validated by RT-qPCR. CaSki cells showed significantly higher glyceride levels than that of C33A cells, as verified by oil red O staining and glyceride assays. The above results showed that the metabolomic differences between C33A and CaSki cells were relatively obvious, especially in lipid metabolism, which might be related to the decreased expression of PPARG and p53 caused by HPV E6. Further studies on the molecular mechanism of lipid metabolism heterogeneity in cervical cancer cell lines with or without HPV could provide a new reference for the development of CC and individualized treatments of tumour patients.

Methods for the rapid screening of group A streptococci: fluorescent in situ hybridization versus immunochromatography.

OBJECTIVE: To evaluate the accuracy of detection for screening group A streptococci (GAS) in pediatric clinics using fluorescent in situ hybridization (FISH) and immunochromatographic assay (ICA). SUBJECTS AND METHODS: A group of 630 children who attended an outpatient clinic with signs and symptoms of acute upper respiratory tract infection were enrolled in this study. Specimens were collected using a double-swab collection device. Culturing methods were employed as the gold standard for comparison. Discordant results (i.e., positive results for FISH or ICA along with negative culture results) were further evaluated by using Todd-Hewitt broth (THB) with subsequent subculture for group A selective streptococcus agar. True-positive or false-positive of GAS was determined by the presence or absence of THB subculture. The diagnostic characteristics of FISH and ICA were assessed. RESULTS: After discrepant analysis, the sensitivity, specificity and positive and negative predictive values were as follows: 89.2, 100, 100 and 95.4%, respectively, for FISH; corresponding values for ICA were 76.8, 98.6, 96.1 and 90.5%. CONCLUSION: The results demonstrated that the FISH assay had a higher detection sensitivity than ICA and is suitable for rapid and accurate GAS screening in pediatric clinics.

Correction: Lymphopenia predicts disease severity of COVID-19: a descriptive and predictive study.

[This corrects the article DOI: 10.1038/s41392-020-0148-4.].

A preliminary study on serological assay for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in 238 admitted hospital patients.

In this study, we aimed to evaluate the diagnostic value of serological assay for SARS-CoV-2. A newly-developed ELISA assay for IgM and IgG antibodies against N protein of SARS-CoV-2 was used to screen the serums of 238 admitted hospital patients between February 6 and February 14, 2020 with confirmed or suspected SARS-CoV-2. SARS-CoV-2 RNA was detected on pharyngeal swab specimens using real time RT-PCR. 194 (81.5%) of the serums were detected to be antibody (IgM and/or IgG) positive, significantly higher than the positive rate of viral RNA (64.3%). There was no difference in the positive rate of antibodies between the confirmed patients (83.0%, 127/153) and the suspected patients (78.8%, 67/85), whose nucleic acid tests were negative. The antibody positive rates were very low in the first five days after initial onset of symptoms, and then rapidly increased as the disease progressed. After 10 days, the antibody positive rates jumped from below 50% to over 80%. However, the positive rates of viral RNA maintained above 60% in the first 11 days after initial onset of symptoms, and then rapidly decreased. Overall, the suspected patients were most likely infected by SARS-CoV-2. Before the 11th day after initial onset of symptoms, nucleic acid test is key for confirmation of viral infection. The combination of serological assay can greatly improve the diagnostic efficacy. After the 11th day post-disease onset, the diagnosis for viral infection should be majorly dependent on serological assay.

Aberrant lncRNA Expression in Multiple Myeloma.

Multiple myeloma (MM), a type of malignant tumor, is characterized by dysplasia of clonal plasma cells in the bone marrow. People with MM will have damaged organs or tissues due to secretion of large amounts of monoclonal immunoglobulin or fragments (M protein). Despite improved survivability by novel treatment strategies over the last decade, MM is still incurable by current therapies. Long noncoding RNAs (lncRNAs), with length of more than 200 nucleotides, have been reported to act as important regulators in many diseases, including MM. Recent studies have reported aberrant lncRNA expression in MM; these dysregulated lncRNAs can play oncogenic and/or tumor-suppressive roles in the development and progression of MM. In this article, we present a general overview on the role of lncRNAs in MM pathogenesis and discuss their potential as prognostic biomarkers and targets for treatment.

[Research Advance in Light Chain Escape of Multiple Myeloma -Review].

More than 80% of patients with MM is present as intact monoclonal immunoglobulin (Ig). Usually, the patients with intact immunoglobulin MM (IIMM) show parallel fluctuations of their intact Ig and FLCs or BJP. In the era of novel agents, including thalidomide, lenalidomide and bortezomib, the natural disease development and classic relapse patterns have been changed, the relapse was characterized by an increase in sFLC or BJP without a corresponding increase in paraprotein level, a phenomenon termed "light chain escape", indicates a worse outcome in patients with MM. This review focuses on the mechanism, clinical significance and early diagnosis of light chain escape.

Risk factors and outcomes of hospitalized patients with blood infections caused by multidrug-resistant Acinetobacter baumannii complex in a hospital of Northern China.

The purpose of this study was to determine the risk factors and outcomes of bloodstream infections caused by multidrug-resistant (MDR) Acinetobacter baumannii complex in a hospital of Northern China. Risk factors associated with MDR A baumannii complex included older age, pneumonia, using drainage catheters, and intensive care unit stay. Multivariate analysis showed that multidrug resistance and mechanical ventilation were identified as independent risk factors for 30-day mortality in patients with A baumannii complex bacteremia.

Analysis of methicillin-resistant Staphylococcus aureus major clonal lineages by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS).

Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen associated with nosocomial infections in many countries. Multilocus sequence typing (MLST) is one of the genetic typing methods used to type MRSA with a high discriminatory power, however, it is labor-intensive, timely, and costly. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) coupled with ClinProTools is a potential tool to discover biomarker peaks and to generate a classification model based on highly sophisticated mathematical algorithms to discriminate clonal lineages. We investigated the performance of MALDI-TOF MS for discriminating 154 MRSA-ST239, 72 MRSA-ST5, 30 MRSA-ST59, 14 MRSA-ST45, and 20 MRSA-OST (other clonal lineages). Our results indicate that the model construction and validation have good potency to discriminate ST45 from other lineages with a sensitivity and a specificity of both 100%, and a sensitivity of 95.80% and a specificity of 94.62% to identify ST239. For Biotyper classification, the sensitivity and specificity were more than of 90% for ST239, ST59 and ST45, whereas only 81.94% sensitivity for ST5. By single-peak analysis, the peaks m/z 4808 and 9614 can correctly discriminate ST45 a sensitivity and a specificity of both 100%; the peak m/z 6554 can also discriminate ST239 with a sensitivity of 91.9% and a specificity of 85.4%. In conclusion, MALDI-TOF MS coupled with ClinProTools has a high detection performance for MRSA typing with obvious advantages of being rapid, highly accurate, and being a low cost in comparison with MLST.

[Gene cloning, expression and polyclonal antibody preparation and application of translocated intimin receptor-cytoskeleton coupling protein].

AIM: To prepare translocated intimin receptor-cytoskeleton coupling protein (TccP) of Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 and its polyclonal antibody. METHODS: TccP was amplified from the genome of EHEC O157:H7 Sakai strain by PCR and used to construct the recombinant prokaryotic expression vector pET28a-TccP. The recombinant vector was transformed into E.coli BL21( DE3) to express the protein in the bacteria under the induction of isopropy-D-thiogalactoside (IPTG). After purification, the protein was injected into New Zealand rabbits to prepare polyclonal antibody. Then the antibody was tested by ELISA and Western blotting for its sensitivity and specificity. The rabbit anti-TccP polyclonal antibody was then applied in the study on the localization of TccP within the host cells adhered by EHEC O157:H7. RESULTS: The sequence of TccP cDNA we amplified was the same as reported by GenBank. The recombinant prokaryotic expression vector pET28a-TccP was constructed successfully. Western blotting revealed that M(r); of the target protein expressed in E.coli BL21(DE3) was 37 000 and the rabbit anti-TccP polyclonal antibody had a specific reaction with the target protein, which demonstrated that the recombinant protein and its polyclonal antibody were prepared successfully. Immunofluorescence detection using rabbit anti-TccP polyclonal antibody showed that TccP aggregated in the cell membrane of the host cell adhered by EHEC O157:H7. CONCLUSION: We successfully prepared the recombinant vector pET28a-TccP and the anti-TccP polyclonal antibody and applied the antibody to confirm the localization of TccP in EHEC O157:H7 adhesion host cells.