Department of Laboratory Medicine, People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, China
Department of Laboratory Medicine, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan
Huang Huayi, School of Laboratory Medicine, Youjiang National Medical University, Baise, Guangxi, 533000, Mindray North America, Mahwah, New Jersey, 07430, USA.
Department of Laboratory Medicine, People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, China
Department of Laboratory Medicine, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan
Huang Huayi, School of Laboratory Medicine, Youjiang National Medical University, Baise, Guangxi, 533000, Mindray North America, Mahwah, New Jersey, 07430, USA.
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In order to better understand the pathological changes of COVID-19, it is conducive to the clinical management of the disease and preparation for the wave of similar pandemics in the future.
The hematological parameters of 52 COVID-19 patients admitted to designated hospitals were retrospectively analyzed. The data was analyzed using SPSS statistical software.
Before treatment, T cell subsets, total lymphocytes, red blood cell distribution width (RDW), eosinophils and basophils were significantly lower than after treatment, while inflammation indicators of neutrophils, neutrophils and lymphocytes The ratio (NLR) and C β-reactive protein (CRP) levels as well as red blood cells (RBC) and hemoglobin decreased significantly after treatment. The T cell subsets, total lymphocytes and basophils of severe and critically ill patients were significantly lower than those of moderate patients. Neutrophils, NLR, eosinophils, procalcitonin (PCT) and CRP are significantly higher in severe and critically ill patients than in moderate patients. The CD3+, CD8+, total lymphocytes, platelets, and basophils of patients over 50 years old are lower than those under 50 years old, while neutrophils, NLR, CRP, RDW in patients over 50 years old are higher than those under 50 years old. In severe and critically ill patients, there is a positive correlation between prothrombin time (PT), alanine aminotransferase (ALT) and aspartate aminotransferase (AST).
T cell subsets, lymphocyte count, RDW, neutrophils, eosinophils, NLR, CRP, PT, ALT and AST are important indicators in management, especially for severe and critically ill patients with COVID-19.
The 2019 Coronavirus Disease (COVID-19) pandemic caused by a new type of coronavirus broke out in December 2019 and spread rapidly across the globe. 1-3 At the beginning of the outbreak, the clinical focus was on manifestations and epidemiology, combined with computed tomography to image patients 4 and 5, and then diagnosed with positive nucleotide amplification results. However, various pathological injuries were later found in different organs. 6-9 More and more evidences show that the pathophysiological changes of COVID-19 are more complicated. The virus attack causes multiple organ damage and the immune system overreacts. Increases in serum and alveolar cytokines and inflammatory response proteins have been observed7, 10-12, and lymphopenia and abnormal T cell subsets have been found in critically ill patients. 13, 14 It is reported that the ratio of neutrophils to lymphocytes has become a useful indicator for distinguishing malignant and benign thyroid nodules in clinical practice. 15 NLR can also help distinguish patients with ulcerative colitis from healthy controls. 16 It also plays a role in thyroiditis and is associated with type 2 diabetes. 17, 18 RDW is a marker of erythrocytosis. Studies have found that it helps to distinguish thyroid nodules, diagnose rheumatoid arthritis, lumbar disc disease, and thyroiditis. 19-21 CRP is a universal predictor of inflammation and has been studied in many cases. 22 It has recently been discovered that NLR, RDW and CRP are also involved in COVID-19 and play an important role in the diagnosis and prognosis of the disease. 11, 14, 23-25 Therefore, the results of laboratory tests are important for evaluating the patient’s condition and making treatment decisions. We retrospectively analyzed the laboratory parameters of 52 COVID-19 patients who were hospitalized in designated hospitals in South China according to their pre- and post-treatment, severity, and age, to further understand the pathological changes of the disease and help future clinical management of COVID-19 .
This study conducted a retrospective analysis of 52 COVID-19 patients admitted to the designated hospital Nanning Fourth Hospital from January 24, 2020 to March 2, 2020. Among them, 45 were moderately ill and 5 were critically ill. For example, the age ranges from 3 months to 85 years old. In terms of gender, there were 27 males and 25 females. The patient has symptoms such as fever, dry cough, fatigue, headache, shortness of breath, nasal congestion, runny nose, sore throat, muscle pain, diarrhea, and myalgia. Computed tomography showed that the lungs were patchy or ground glass, indicating pneumonia. Diagnose according to the 7th edition of the Chinese COVID-19 Diagnosis and Treatment Guidelines. Confirmed by real-time qPCR detection of viral nucleotides. According to the diagnostic criteria, patients were divided into moderate, severe, and critical groups. In moderate cases, the patient develops fever and respiratory syndrome, and imaging findings show pneumonia patterns. If the patient meets any of the following criteria, the diagnosis is severe: (a) respiratory distress (breathing rate ≥30 breaths/min); (b) resting finger blood oxygen saturation ≤93%; (c) arterial oxygen pressure (PO2) )/Inspiratory fraction O2 (Fi O2) ≤300 mm Hg (1 mm Hg = 0.133 kPa). If the patient meets any of the following criteria, the diagnosis is severe: (a) respiratory failure that requires mechanical ventilation; (b) shock; (c) other organ failure that requires treatment in the intensive care unit (ICU). According to the above criteria, 52 patients were diagnosed as severely ill in 2 cases, severely ill in 5 cases, and moderately ill in 45 cases.
All patients, including moderate, severe and critically ill patients, are treated in accordance with the following basic procedures: (a) General adjuvant therapy; (b) Antiviral therapy: lopinavir/ritonavir and α-interferon; (c) The dosage of traditional Chinese medicine formula can be adjusted according to the patient’s condition.
This study was approved by the Review Committee of the Research Institute of Nanning Fourth Hospital and was used to collect patient information.
Peripheral blood hematology analysis: routine hematology analysis of peripheral blood is performed on Mindray BC-6900 hematology analyzer (Mindray) and Sysmex XN 9000 hematology analyzer (Sysmex). The fasting ethylenediaminetetraacetic acid (EDTA) anticoagulant blood sample was collected the morning after the patient was admitted to the hospital. The consistency assessment between the above two blood analyzers was verified in accordance with laboratory quality control procedures. In hematology analysis, white blood cell (WBC) count and differentiation, red blood cell (RBC) and index are obtained together with scatter plots and histograms.
Flow cytometry of T lymphocyte subpopulations: BD (Becton, Dickinson and Company) FACSCalibur flow cytometer was used for flow cytometry analysis to analyze T cell subpopulations. Analyze the data by MultiSET software. The measurement was carried out in accordance with standard operating procedures and the manufacturer’s instructions. Use an EDTA anticoagulated blood collection tube to collect 2 ml of venous blood. Gently mix the sample by turning the sample tube several times to prevent condensation. After the sample is collected, it is sent to the laboratory and analyzed within 6 hours at room temperature.
Immunofluorescence analysis: C-reactive protein (CRP) and procalcitonin (PCT) were analyzed immediately after the completion of the analysis using blood samples analyzed by hematology, and analyzed on the FS-112 immunofluorescence analyzer (Wondfo Biotech Co., LTD .) on the analysis. ) Follow the manufacturer’s instructions and laboratory procedure standards.
Analyze serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) on the HITACHI LABOSPECT008AS chemical analyzer (HITACHI). The prothrombin time (PT) was analyzed on the STAGO STA-R Evolution analyzer (Diagnostica Stago).
Reverse transcription quantitative polymerase chain reaction (RT-qPCR): Use RNA templates isolated from nasopharyngeal swabs or lower respiratory tract secretions to perform RT-qPCR to detect SARS-CoV-2. Nucleic acids were separated on the SSNP-2000A nucleic acid automatic separation platform (Bioperfectus Technologies). The detection kit was provided by Sun Yat-sen University Daan Gene Co., Ltd. and Shanghai BioGerm Medical Biotechnology Co., Ltd. The thermal cycle was performed on an ABI 7500 thermal cycler (Applied Biosystems). Viral nucleoside test results are defined as positive or negative.
SPSS version 18.0 software was used for data analysis; paired-sample t-test, independent-sample t-test, or Mann-Whitney U test were applied, and a P value <.05 was considered significant.
Five critically ill patients and two critically ill patients were older than those in the moderate group (69.3 vs. 40.4). The detailed information of 5 critically ill and 2 critically ill patients are shown in Tables 1A and B. Severe and critically ill patients are usually low in T cell subsets and total lymphocyte counts, but the white blood cell count is roughly normal, except for patients with elevated white blood cells (11.5 × 109/L). Neutrophils and monocytes are also usually high. The serum PCT, ALT, AST and PT values of 2 critically ill patients and 1 critically ill patient were high, and the PT, ALT, AST of 1 critically ill patient and 2 critically ill patients were positively correlated. Almost all 7 patients had high CRP levels. Eosinophils (EOS) and basophils (BASO) tend to be low in critically ill and critically ill patients (Table 1A and B). Table 1 lists the description of the normal range of hematological parameters in the Chinese adult population.
Statistical analysis showed that before treatment, CD3+, CD4+, CD8+ T cells, total lymphocytes, RBC distribution width (RDW), eosinophils and basophils were significantly lower than after treatment (P = .000,. 000, .000, .012, .04, .000 and .001). The inflammatory indicators neutrophils, neutrophil/lymphocyte ratio (NLR) and CRP before treatment were significantly higher than after treatment (P = .004, .011 and .017, respectively). Hb and RBC decreased significantly after treatment (P = .032, .026). PLT increased after treatment, but it was not significant (P = .183) (Table 2).
The T cell subsets (CD3+, CD4+, CD8+), total lymphocytes and basophils of severe and critically ill patients were significantly lower than those of moderate patients (P = .025, 0.048, 0.027, 0.006 and .046). The levels of neutrophils, NLR, PCT and CRP in severe and critically ill patients were significantly higher than those in moderate patients (P = .005, .002, .049 and .002, respectively). Severe and critically ill patients had lower PLT than moderate patients; however, the difference was not statistically significant (Table 3).
The CD3+, CD8+, total lymphocytes, platelets, and basophils of patients over 50 years old were significantly lower than those of patients under 50 years old (P = .049, 0.018, 0.019, 0.010 and .039, respectively), while those over 50 years old Patients’ neutrophils, NLR ratio, CRP levels and RDW were significantly higher than those of patients under 50 years of age (P = .0191, 0.015, 0.009, and .010, respectively) (Table 4).
COVID-19 is caused by infection with the coronavirus SARS-CoV-2, which first appeared in Wuhan, China in December 2019. The SARS-CoV-2 outbreak spread rapidly afterwards and led to a global pandemic. 1-3 Due to limited knowledge of the epidemiology and pathology of the virus, the mortality rate at the beginning of the outbreak is high. Although there are no antiviral drugs, the follow-up management and treatment of COVID-19 has been greatly improved. This is especially true in China when adjuvant therapies are combined with traditional Chinese medicine to treat early and moderate cases. 26 COVID-19 patients have benefited from a better understanding of the pathological changes and laboratory parameters of the disease. disease. Since then, the mortality rate has declined. In this report, there were no deaths among the 52 cases analyzed, including 7 severe and critically ill patients (Table 1A and B).
Clinical observations have found that most patients with COVID-19 have reduced lymphocytes and T cell subpopulations, which are related to the severity of the disease. 13, 27 In this report, it was found that CD3+, CD4+, CD8+ T cells, total lymphocytes, RDW before treatment, eosinophils and basophils were significantly lower than after treatment (P = .000, .000, .000, .012, .04, .000 and .001). Our results are similar to previous reports. These reports have clinical significance in monitoring the severity of COVID-19.8, 13, 23-25, 27, while the inflammatory indicators neutrophils, neutrophils/lymphocyte ratio (NLR ) And CRP after pre-treatment than treatment (P = .004, .011 and .017, respectively), which have been noticed and reported previously in COVID-19 patients. Therefore, these parameters are considered to be useful indicators for the treatment of COVID-19.8. After treatment, 11 hemoglobin and red blood cells were significantly reduced (P = .032, 0.026), indicating that the patient had anemia during the treatment. An increase in PLT was observed after treatment, but it was not significant (P = .183) (Table 2). The decrease in lymphocytes and T cell subpopulations is thought to be related to cell depletion and apoptosis when they accumulate in inflammatory sites that fight the virus. Or, they may have been consumed by excessive secretion of cytokines and inflammatory proteins. 8, 14, 27-30 If the lymphocyte and T cell subsets are persistently low and the CD4+/CD8+ ratio is high, the prognosis is poor. 29 In our observation, lymphocytes and T cell subsets recovered after treatment, and all 52 cases were cured (Table 1). High levels of neutrophils, NLR, and CRP were observed before treatment, and then significantly decreased after treatment (P = .004, .011, and .017, respectively) (Table 2). The function of T cell subsets in infection and immune response has been previously reported. 29, 31-34
As the number of severe and critically ill patients is too small, we did not perform statistical analysis on the parameters between severe and critically ill patients and moderate patients. The T cell subsets (CD3+, CD4+, CD8+) and total lymphocytes of severe and critically ill patients are significantly lower than those of moderate patients. The levels of neutrophils, NLR, PCT, and CRP in severe and critically ill patients were significantly higher than those in moderate patients (P = .005, .002, .049, and .002, respectively) (Table 3). Changes in laboratory parameters are related to the severity of COVID-19.35. 36 The cause of basophilia is unclear; this may be due to the consumption of food while fighting the virus at the site of infection similar to lymphocytes. 35 The study found that patients with severe COVID-19 also had reduced eosinophils; 14 However, our data did not show that this phenomenon may be due to the small number of severe and critical cases observed in the study.
Interestingly, we found that in severe and critically ill patients, there is a positive correlation between PT, ALT, and AST values, indicating that multiple organ damage occurred in the virus attack, as mentioned in other observations. 37 Therefore, they may be new useful parameters for evaluating the response and prognosis of COVID-19 treatment.
Further analysis showed that the CD3+, CD8+, total lymphocytes, platelets and basophils of patients over 50 years old were significantly lower than those of patients under 50 years old (P = P = .049, .018, .019, .010 and. 039, respectively), while the levels of neutrophils, NLR, CRP, and RBC RDW in patients over 50 years of age were significantly higher than those of patients under 50 years of age (P = .0191, 0.015, 0.009, and .010, respectively) (Table 4) . These results are similar to previous reports. 14, 28, 29, 38-41 Decrease in T cell subpopulations and high CD4+/CD8+ T cell ratios are related to disease severity; elderly cases tend to be more severe; therefore, more lymphocytes will be consumed in the immune response or Seriously damaged. Likewise, a higher RBC RDW indicates that these patients have developed anemia.
Our research results further confirm that hematological parameters are of great significance for better understanding of the clinicopathological changes of COVID-19 patients and for improving the guidance of treatment and prognosis.
Liang Juanying and Nong Shaoyun collected data and clinical information; Jiang Liejun and Chi Xiaowei performed data analysis; Dewu Bi, Jun Cao, Lida Mo, and Xiaolu Luo performed routine analysis; Huang Huayi was responsible for the conception and writing.
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Post time: Jul-22-2021