Glucocorticoids and the relationship between glucocorticoids and viral load, Omicron wave in mainland China | Journal of Virology

Study Population and Design

From December 2022 to January 2023, patients diagnosed with COVID-19 infection and admitted to the emergency department of Beijing United Medical College Hospital participated in this prospective study. All patients signed informed consent, and the study was approved by the hospital ethics committee. This study was approved by the PUMCH Ethics Committee and registered at Chictr.org.cn (identifier CHICTR2000030349). Inclusion criteria were (1) duration of hospitalization ≥48h, (2) confirmed severe and critical Covid-19 infection. Exclusion criteria were (1) confirmed COVID-19 infection in mild and moderate patients, and (2) all patients using antiviral drugs during illness, including before visiting a doctor. According to the diagnostic and treatment protocols for Covid-19 (9th and 10th edition) established by the National Health Commission of the People’s Republic of China, an infection with Covid-19 is defined as having (1) one of the following clinical manifestations of COVID-19 (2) or multiple etiological and serological test results: (a) positive covid-19 nucleic acid test, (b) positive covid-19 antigen, (c) positive covid-19-19 isolation and culture, and (d)) in recovery In the mid-term, the level of novel coronavirus-specific immunoglobulin G antibodies was equal to or four times greater than in the acute phase. Clinical categories are defined as follows. “Mild” refers to mild manifestations of the main clinical symptoms of upper respiratory tract infection, such as dry throat, sore throat, cough and fever. “Moderate” refers to persistent high fever > 3 days or cough, or both with respiratory rate (RR) < 30 breaths/min/min when breathing air at rest<30次，氧饱和度> 93%, and imaging shows that. Characteristic manifestations of 19009 pneumonia.A “severe” classification was used for adults who met any of the following criteria that could not be explained by anything other than COVID-19 infection: (a) shortness of breath (i.e., RR ≥ 30 breaths/min), (b) ) Oxygen saturation<93％时，当静止呼吸空气时，（c）动脉部分氧气压力/吸收氧气≤300mmHg（1 mm Hg = 0.133 kPa），（d）逐渐恶化临床症状和肺部成像，表明病变的病变在24至48小时内取得了显着进步（即，> 50%). Critical patients are defined as patients who develop one of the following conditions: (1) respiratory failure requiring mechanical ventilation; (2) shock; (3) complicated by other organ failure requiring ICU care.

All enrolled patients were labeled as early (8–14 days), recovered (8–14 days), and late (>14 days) based on symptoms (fever, sore throat, cough, and dyspnea) or dyspnea (0–7 days) ) and late (>14 days) the time of their first positive COVID-19 nucleic acid test. Since the criteria for glucocorticoids are severe and important patients with progressive deterioration of oxygenation index, rapid imaging progress, and excessive activation of the body’s inflammatory response as established in the Diagnosis and Treatment Protocol for COVID-19 (9th Edition). After enrollment in the People’s Republic of China, the need for glucocorticoids, administered orally at a dose of 5 mg dexamethasone daily, was assessed by at least two experienced clinicians.

clinical and laboratory assessment

On the day of hospitalization, all registered patients underwent comprehensive clinical and laboratory evaluation. Information recorded includes age, sex, underlying disease, important biochemical indicators of infection (e.g., procalcitin, creatinine, albumin, bilirubin, and coagulation), Acute Physiology and Chronic Health Evaluation II (Apache-II), and sequential organ failure assessment (SOFA) (15) score, and documentation of any life-sustaining treatment lasting ≥24 hours (e.g., mechanical ventilation, noninvasive positive pressure ventilation, high-flow nasal cannula, Venturi membrane, and extracorporeal membrane oxygenation, accelerator oxidant drugs or renal replacement therapy).

Immunology laboratory tests

Serum was obtained from all patients to test immune parameters including white blood cell count, complement, immunoglobulins, inflammatory factors, T cell subsets and ferritin. Briefly, detection of T cell subsets begins with the isolation of peripheral blood mononuclear cells, which are then stained with different combinations of fluorescent monoclonal antibodies. Finally, T cells (CD3⁺), CD4⁺ T cell subset (CD4⁺CD3⁺ and CD28⁺CD4⁺), CD8⁺ T cell subset (CD8⁺CD3⁺CD28⁺CD8⁺), B cells (CD19⁺) and NK cells (CD3^–CD16⁺CD56⁺Detected using flow cytometry).

RNA extraction and reverse transcription-polymerase chain reaction (RT-PCR) detection of 19 viral payloads with the HNA signature

Throat swabs are taken daily from all admitted patients for 19 viral load tests. Instruct the patient to rinse their mouth with water and then insert the swab into their mouth and pass it through their tongue. We rub back and forth at least three times on both sides of the pharyngeal tonsils, and then we rub at least three times on the posterior pharyngeal wall. The swab is withdrawn and placed into a collection tube. Collect all swabs and soak them in 1,000 μl phosphate buffered saline. After shaking for 30 s, 400 μl of the sample was taken out to extract nucleic acids using SY619 nucleic acid extraction kit (Suzhou Xinbo Biotechnology Co. Ltd., Suzhou, China). The number of viral copies is measured using RT-PCR using primers and probes targeting the N gene of severe acute respiratory syndrome coronavirus 2 (SARS-COV-2).Dilute reference standard from 1×10⁸ The copy is 1×10⁹ copy. PCR amplification cycles were 15 min at 50°C, 3 min at 95°C, 15 s at 95°C, and 45 s at 60°C, followed by plate reading at 50 cycles. The amplification process, fluorescence signal detection, data storage and analysis were completed by quantitative fluorescence PCR and its software Bio-Rad CFX Manager (Bio-Rad Laboratories Co. Ltd., California, USA). The number of virus copies was calculated based on the standard curve and then converted to log 10 for statistical analysis. Negative transition of COVID-19 is defined as two consecutive cycle threshold (CT) values ​​of SARS-COV-2 nucleic acid nitrogen

genes and open reading frame Gene ≥35.

Statistical Analysis

All analyzes were performed using SPSS for Windows version 24.0 (IBM Corp., Armonk, NY, USA). Kolmogorov-Smirnov test, Student t-Test and Mann-Whitney U test were used to examine the cumulative distribution function and analyze normally distributed continuous and non-parametric variables. Choose chi-square or Fisher’s exact test, appropriate for comparing categorical variables. p-values ​​associated with “not assumed to be equal” are reported for variables that violate the homogeneity of variance assumption. All tests performed were two-tailed, and P < 0.05 was considered statistically significant.