Recent advances in non-optical microfluidic platforms for bioparticle detection.

The effective analysis of the basic structure and functional information of bioparticles are of great significance for the early diagnosis of diseases. The synergism between microfluidics and particle manipulation/detection technologies offers enhanced system integration capability and test accuracy for the detection of various bioparticles. Most microfluidic detection platforms are based on optical strategies such as fluorescence, absorbance, and image recognition. Although optical microfluidic platforms have proven their capabilities in the practical clinical detection of bioparticles, shortcomings such as expensive components and whole bulky devices have limited their practicality in the development of point-of-care testing (POCT) systems to be used in remote and underdeveloped areas. Therefore, there is an urgent need to develop cost-effective non-optical microfluidic platforms for bioparticle detection that can act as alternatives to optical counterparts. In this review, we first briefly summarise passive and active methods for bioparticle manipulation in microfluidics. Then, we survey the latest progress in non-optical microfluidic strategies based on electrical, magnetic, and acoustic techniques for bioparticle detection. Finally, a perspective is offered, clarifying challenges faced by current non-optical platforms in developing practical POCT devices and clinical applications.

Causal Associations Between Blood Lipids and COVID-19 Risk: A Two-Sample Mendelian Randomization Study.

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Analysis of Peripheral Blood IL-6 and Leukocyte Characteristics in 364 COVID-19 Patients of Wuhan.

SARS-CoV-2, the pathogen of COVID-19, is spreading around the world. Different individuals infected with COVID-19 have different manifestations. It is urgent to determine the risk factors of disease progress of COVID-19. 364 patients diagnosed with COVID-19, who were admitted to Wuhan Pulmonary Hospital from February 3, 2020 to March 16, 2020, were divided into mild, ordinary, severe, and critical groups, according to Chinese novel coronavirus pneumonia diagnosis and treatment plan. Peripheral blood IL-6 and leukocyte characteristics were analyzed, to evaluate the correlation with the severity of COVID-19. The levels of peripheral blood IL-6 were 2.35 ± 0.46 pg/ml (mild), 6.48 ± 1.13 pg/ml (ordinary), 20.30 ± 5.15 pg/ml (severe), and 123.48 ± 44.31 pg/ml (critical). The leukocytes were 5.70 ± 0.41×109/L (mild), 6.21 ± 0.14×109/L (ordinary), 6.37 ± 0.26×109/L (severe), and 10.03 ± 1.43×109/L (critical). The lymphocytes were 1.46 ± 0.19×109/L (mild), 1.89 ± 0.14×109/L (ordinary), 1.26 ± 0.07×109/L (severe), and 1.17 ± 0.23×109/L (critical). The neutrophils were 3.63 ± 0.36×109/L (mild), 3.78 ± 0.11×109/L (ordinary), 4.47 ± 0.25×109/L (severe), and 7.92 ± 1.19×109/L (critical). The monocytes were 0.42 ± 0.05×109/L (mild), 0.44 ± 0.01×109/L (ordinary), 0.46 ± 0.02×109/L (severe), and 0.78 ± 0.25×109/L (critical). Conclusively, increase of peripheral blood IL-6 and decrease of lymphocytes can be used as the indicators of severe COVID-19. The increase of neutrophils and monocytes was noticed in critical cases of COVID-19, suggesting that the increase of neutrophils and monocytes should be considered as risk factors of critical cases of COVID-19. Peripheral blood IL-6 and leukocyte characteristics were also analyzed in different age groups. The increase of serum IL-6, decrease of lymphocytes, and increase of neutrophils were noticed in patients over 60 years old.

Neutrophil-to-lymphocyte ratio, a critical predictor for assessment of disease severity in patients with COVID-19.

INTRODUCTION: Monitoring of laboratory indicators is important for predicting changes in disease severity and clinical outcomes. We aimed to identify the critical predictors that can effectively assess the disease conditions of patients with COVID-19 by analyzing the clinical characteristics and laboratory findings of patients with SARS-CoV-2 infection. METHODS: All consecutive patients (n = 294) with confirmed SARS-CoV-2 infection admitted to the General Hospital of Central Theater Command of the PLA from February 6 to February 21, 2020, were enrolled. These patients were divided into the severe group and the nonsevere group according to disease severity during hospitalization. RESULTS: The median neutrophil-to-lymphocyte ratio (NLR) value of the severe patients was dramatically higher than that of the nonsevere patients (10.4 vs 2.6; P < .001). The NLR value equal to 5 was a boundary value worthy of reference, because more than 80% severe patients had an NLR value greater than 5 and over 80% nonsevere patients had an NLR value less than 5. The NLR value of these COVID-19 patients was positively and respectively correlated with the values of C-reactive protein (R = .5921, P < .001), lactate dehydrogenase (R = .4509, P < .001), procalcitonin (R = .5504, P < .001), fibrinogen (R = .4710, P < .001), and D-dimers (R = .4425, P < .001). However, the NLR value was merely and positively correlated with the interleukin-6 value (R = .3594, P < .05), but had no correlations with the values of interleukin-10, interleukin-4, interleukin-17, interferon-γ, and tumor necrosis factor-α (P > .05). DISCUSSION: Neutrophil-to-lymphocyte ratio is a critical predictor for assessment of disease severity in patients with COVID-19, and it has a close relation with the laboratory indicators related to disease conditions.

Clinical application of Chemiluminescence Microparticle Immunoassay for SARS-CoV-2 infection diagnosis.

BACKGROUND: The unsatisfactory accuracy and capacity of real time RT-PCR depends on several unavoidable reasons, which cannot meet the demands for COVID-19 diagnosis. METHODS: 206 serum samples were collected from patients who were treated in the General Hospital of the Central Theater Command of the PLA between January 18 and April 4, 2020. 270 serum samples from healthy blood donors were used as control. IgM and total antibodies (Ab) against SARS-CoV-2 were detected by Chemiluminescence Microparticle Immunoassay (CMIA). RESULTS: Among the 206 patients, the positive rate of IgM and Ab were 149/206 (72.3 %) and 187/206 (90.8 %), respectively. And the specificity of IgM and Ab detection were 99.3 % and 98.9 %, respectively. The sensitivity of CMIA for Ab detection was significantly higher than that of IgM. An increase of the positive rate and S/CO value for detecting IgM and Ab accompanied with the increasing of days post-disease onset (d.p.o.) were observed. The positive rate of Ab detected by CMIA increased rapidly after 7 d.p.o., while that of IgM was obviously increased after 14 d.p.o.. In addition, the age and gender of these patients did not affect the seroconversion and titer of antibodies during the whole course. The disease-severity of patients had no effect on the seroconversion of antibodies. However, the critical patients possessed a much higher antibody titers than the no-critical cases after 14 d.p.o.. CONCLUSIONS: The CMIA can provide important complementation to nucleic acid assay and help to enhance the accuracy and capacity of diagnosis of SARS-CoV-2 infection.

[Large- scale prospective clinical study on prophylactic intervention of COVID-19 in community population using Huoxiang Zhengqi Oral Liquid and Jinhao Jiere Granules].

To scientifically evaluate the intervention effect of Chinese medicine preventive administration(combined use of Huo-xiang Zhengqi Oral Liquid and Jinhao Jiere Granules) on community population in the case of coronavirus disease 2019(COVID-19), a large cohort, prospective, randomized, and parallel-controlled clinical study was conducted. Total 22 065 subjects were included and randomly divided into 2 groups. The non-intervention group was given health guidance only, while the traditional Chinese medicine(TCM) intervention group was given two coordinated TCM in addition to health guidance. The medical instructions were as follows. Huoxiang Zhengqi Oral Liquid: oral before meals, 10 mL/time, 2 times/day, a course of 5 days. Jinhao Jiere Granules: dissolve in boiling water and take after meals, 8 g/time, 2 times/day, a course of 5 days, followed up for 14 days, respectively. The study found that with the intake of medication, the incidence rate of TCM intervention group was basically maintained at a low and continuous stable level(0.01%-0.02%), while the non-intervention group showed an overall trend of continuous growth(0.02%-0.18%) from 3 to 14 days. No suspected or confirmed COVID-19 case occurred in either group. There were 2 cases of colds in the TCM intervention group and 26 cases in the non-intervention group. The incidence of colds in the TCM intervention group was significantly lower(P<0.05) than that in the non-intervention group. In the population of 16-60 years old, the incidence rate of non-intervention and intervention groups were 0.01% and 0.25%, respectively. The difference of colds incidence between the two groups was statistically significant(P<0.05). In the population older than 60 years old, they were 0.04% and 0.21%, respectively. The incidence of colds in the non-intervention group was higher than that in the intervention group, but not reaching statistical difference. The protection rate of TCM for the whole population was 91.8%, especially for the population of age 16-60(95.0%). It was suggested that TCM intervention(combined use of Huoxiang Zhengqi Oral Liquid and Jinhao Jiere Granules) could effectively protect community residents against respiratory diseases, such as colds, which was worthy of promotion in the community. In addition, in terms of safety, the incidence of adverse events and adverse reactions in the TCM intervention group was relatively low, which was basically consistent with the drug instructions.

[Analysis of potential role of Chinese classic prescriptions in treatment of COVID-19 based on TCMATCOV platform].

With the global outbreak of coronavirus disease 2019(COVID-19), screening of effective drugs has became the emphasis of research today; furthermore, screening of Chinese classic prescriptions has became one of the directions for drug development. This study analyzed the application of classic prescriptions in the diagnosis and treatment schemes based on the Diagnosis and Treatment Schemes for Coronavirus Disease at the country, provincial and municipal levels, and further explored its disrobing effect on COVID-19 disease severe phase network, and selected representative prescriptions for core target screening and gene enrichment analysis, so as to reveal its mechanism of action. Among them, 13 prescriptions were found to be used for 10 times or more, including Maxing Shigan Tang, Yinqiao San, Shengjiang San, Dayuan Drink, Xuanbai Chengqi Decoction. In addition, the COVID-19 efficacy prediction analysis platform(TCMATCOV platform) was used to calculate the network disturbances of the Chinese classic prescriptions involved. Based on the prediction results, 68 classic prescriptions were assessed on the COVID-19 disease network robustness disturbance. The average disturbance scores for the interaction confidence scores were ranked to be 0.4, 0.5, and 0.6 from the highest to the lowest. There were 7 prescriptions with a score of 17 or more, and 50 prescriptions with a score of 13 or more. Among them, the top three prescriptions were Ganlu Xiaodu Dan(18.19), Lengxiao Wan(17.74), and Maxing Shigan Tang(17.62). After further mining the action targets of these three prescriptions, it was found that COVID-19 disease-specific factors Ccl2, IL10, IL6 and TNF were all the targets of three prescriptions. Through the enrichment analysis of the biological processes of the core targets, it was found that the three prescriptions may prevent the development of the disease by affecting cell-to-cell adhesion, cytokine-mediated signaling pathway, and chronic inflammatory responses to COVID-19 at the severe phase. This study showed that the TCMATCOV platform could evaluate the disturbance effect of different prescriptions on the COVID-19 disease network, and predict potential effectiveness based on the robustness of drug-interfered pneumonia disease networks, so as to provide a reference for further experiments or clinical verification.

Evaluation of Nucleocapsid and Spike Protein-Based Enzyme-Linked Immunosorbent Assays for Detecting Antibodies against SARS-CoV-2.

At present, PCR-based nucleic acid detection cannot meet the demands for coronavirus infectious disease (COVID-19) diagnosis. Two hundred fourteen confirmed COVID-19 patients who were hospitalized in the General Hospital of Central Theater Command of the People's Liberation Army between 18 January and 26 February 2020 were recruited. Two enzyme-linked immunosorbent assay (ELISA) kits based on recombinant severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (rN) and spike protein (rS) were used for detecting IgM and IgG antibodies, and their diagnostic feasibility was evaluated. Among the 214 patients, 146 (68.2%) and 150 (70.1%) were successfully diagnosed with the rN-based IgM and IgG ELISAs, respectively; 165 (77.1%) and 159 (74.3%) were successfully diagnosed with the rS-based IgM and IgG ELISAs, respectively. The positive rates of the rN-based and rS-based ELISAs for antibody (IgM and/or IgG) detection were 80.4% and 82.2%, respectively. The sensitivity of the rS-based ELISA for IgM detection was significantly higher than that of the rN-based ELISA. We observed an increase in the positive rate for IgM and IgG with an increasing number of days post-disease onset (d.p.o.), but the positive rate of IgM dropped after 35 d.p.o. The positive rate of rN-based and rS-based IgM and IgG ELISAs was less than 60% during the early stage of the illness, 0 to 10 d.p.o., and that of IgM and IgG was obviously increased after 10 d.p.o. ELISA has a high sensitivity, especially for the detection of serum samples from patients after 10 d.p.o., so it could be an important supplementary method for COVID-19 diagnosis.

[TCMATCOV--a bioinformatics platform to predict efficacy of TCM against COVID-19].

There is urgent need to discover effective traditional Chinese medicine(TCM) for treating coronavirus disease 2019(COVID-19). The development of a bioinformatic tool is beneficial to predict the efficacy of TCM against COVID-19. Here we deve-loped a prediction platform TCMATCOV to predict the efficacy of the anti-coronavirus pneumonia effect of TCM, based on the interaction network imitating the disease network of COVID-19. This COVID-19 network model was constructed by protein-protein interactions of differentially expressed genes in mouse pneumonia caused by SARS-CoV and cytokines specifically up-regulated by COVID-19. TCMATCOV adopted quantitative evaluation algorithm of disease network disturbance after multi-target drug attack to predict potential drug effects. Based on the TCMATCOV platform, 106 TCM were calculated and predicted. Among them, the TCM with a high disturbance score account for a high proportion of the classic anti-COVID-19 prescriptions used by clinicians, suggesting that TCMATCOV has a good prediction ability to discover the effective TCM. The five flavors of Chinese medicine with a disturbance score greater than 1 are mainly spicy and bitter. The main meridian of these TCM is lung, heart, spleen, liver, and stomach meridian. The TCM related with QI and warm TCM have higher disturbance score. As a prediction tool for anti-COVID-19 TCM prescription, TCMATCOV platform possesses the potential to discovery possible effective TCM against COVID-19.

[Treatment of COVID-19 guided by holistic view of traditional Chinese medicine--therapy aimed at both viral and host].

The global outbreak of coronavirus disease 2019(COVID-19) has further spread, and there is an increasing number of confirmed cases in many countries. On February 28, 2020 of Geneva time, the World Health Organization has raised global risk level to the very high level in view of outbreak of COVID-19. Since some patients' condition appeared to deteriorate rapidly after infection of this 2019 novel coronavirus(2019-nCoV), a variety of treatments should be considered. Holistic view and syndrome differentiation are the two characteristics of traditional Chinese medicine(TCM). Therefore, under the guidance of the holistic view, syndrome diffe-rentiation of TCM has achieved good effects in the treatment of COVID-19. This treatment mainly aimed at eliminating pathogens and strengthening overall health, regulating the balance of body and coordinating various of functions of Zangfu organs. In addition, modern medical proposes host-directed therapy(HDT), a strategy aims to interfere with host cell mechanism, enhance immune responses, and reduce exacerbated inflammation. To some extent, the combined application of HDT and antiviral therapy is highly consistent with the therapeutic concept of the holistic view of TCM. Therefore, under the guidance of the holistic view, syndrome differentiation of TCM uses treatments, such as clearing heat, detoxification, relieving asthma, clearing damp and phlegm, together with Lianhua Qingwen Capsules, Maxing Shigan Decoction, and Haoqin Qingdan Decoction under the guidance of these therapeutic methods. These therapeutic methods and prescriptions intervened with both virus and host at the same time in the treatment of COVID-19, which has important implications for the effective clinical treatment of COVID-19.

[Intervention mechanism of Qingwen Baidu Yin on cytokine storm based on network pharmacology].

The aim of this paper was to explore the intervention mechanism of Qingwen Baidu Yin in cytokine storm based on network pharmacology. TCMSP and TCMIP V2.0 server were used to predict all chemical components and action targets of Qingwen Baidu Yin. Diseases that could be treated by Qingwen Baidu Yin were predicted through Enrichr database. A compound target interaction(PPI) network diagram was constructed using STRING 11.0. OmicShare was used to analyzed the gene ontology(GO) enrichment and enrichment of the Kyoto encyclopedia of genes and genomes(KEGG) pathway of core targets. Component-target-path network diagram was constructed with Cytoscape 3.6.0 software. After analysis of the database, 267 compounds were screened for Qingwen Baidu Yin, involving 1 450 targets, and a protein interaction network was constructed. Total 219 core target proteins were predicted, such as NFKB1, STAT1, RAF1, IL2, JAK1, IL6, TNF, BCL2 and other important targets, and 221 core target pathways were enriched, including cancer pathway, Kaposi's sarcoma-associated herpes virus infection, chemokine signal pathway, PI3 K-AKT signal pathway, EB virus infection, virus carcinogenesis and T cell receptor signaling pathways, a collection of which were highly related to cytokine storms. GO annotation analysis suggested that Qingwen Baidu Yin Decoction may exert therapeutic effects by regulating protein phosphorylation, cell response to cytokine stimulation, cell proliferation, inflammatory response, transmembrane receptor protein tyrosine kinase signaling pathway, and cytokine-mediated signaling pathways. This study revealed potential active components of Qingwen Baidu Yin in defending against cytokine storm and its possible mechanism of action, and provided theoretical basis and technical support for further clinical application of this prescription.

Coronavirus disease 2019 (COVID-19): current status and future perspectives.

Coronavirus disease 2019 (COVID-19) originated in the city of Wuhan, Hubei Province, Central China, and has spread quickly to 72 countries to date. COVID-19 is caused by a novel coronavirus, named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) [previously provisionally known as 2019 novel coronavirus (2019-nCoV)]. At present, the newly identified SARS-CoV-2 has caused a large number of deaths with tens of thousands of confirmed cases worldwide, posing a serious threat to public health. However, there are no clinically approved vaccines or specific therapeutic drugs available for COVID-19. Intensive research on the newly emerged SARS-CoV-2 is urgently needed to elucidate the pathogenic mechanisms and epidemiological characteristics and to identify potential drug targets, which will contribute to the development of effective prevention and treatment strategies. Hence, this review will focus on recent progress regarding the structure of SARS-CoV-2 and the characteristics of COVID-19, such as the aetiology, pathogenesis and epidemiological characteristics.