Viral loads, lymphocyte subsets and cytokines in asymptomatic, mildly and critical symptomatic patients with SARS-CoV-2 infection: a retrospective study.

BACKGROUND: Tens of million cases of coronavirus disease-2019 (COVID-19) have occurred globally. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) attacks the respiratory system, causing pneumonia and lymphopenia in infected individuals. The aim of the present study is to investigate the laboratory characteristics of the viral load, lymphocyte subset and cytokines in asymptomatic individuals with SARS-CoV-2 infection in comparison with those in symptomatic patients with COVID-19. METHODS: From January 24, 2020, to April 11, 2020, 48 consecutive subjects were enrolled in this study. Viral loads were detected by RT-PCR from throat-swab, sputum and feces samples. Lymphocyte subset levels of CD3 + , CD4 + , and CD8 + T lymphocytes, B cells and NK cells were determined with biological microscope and flow cytometric analysis. Plasma cytokines (IL2, IL4, IL5, IL6, IL8, IL10, TNF-α, IFN-α and IFN-γ) were detected using flow cytometer. Analysis of variance (ANOVA), Chi-square or Fisher's exact test and Pearson's Correlation assay was used for all data. RESULTS: Asymptomatic (AS), mild symptoms (MS) and severe or critical cases (SCS) with COVID-19 were 11 (11/48, 22.9%), 26 (54.2%, 26/48) and 11 cases (11/48, 22.9%), respectively. The mean age of AS group (47.3 years) was lower than SCS group (63.5 years) (P < 0.05). Diabetes mellitus in AS, MS and SCS patients with COVID-19 were 0, 6 and 5 cases, respectively, and there was a significant difference between AS and SCS (P < 0.05). No statistical differences were found in the viral loads of SARS-CoV-2 between AS, MS and SCS groups on admission to hospital and during hospitalization. The concentration of CD 3 + T cells (P < 0.05), CD3 + CD4 + T cells (P < 0.05), CD3 + CD8 + T cells (P < 0.01), and B cells (P < 0.05) in SCS patients was lower than in AS and MS patients, while the level of IL-5 (P < 0.05), IL-6 (P < 0.05), IL-8 (P < 0.01) and IL-10 (P < 0.01), and TNF-α (P < 0.05) was higher. The age was negatively correlated with CD3 + T cells (P < 0.05), CD3 + CD4 + T cells (P < 0.05), and positively correlated with IL-2 (P < 0.001), IL-5 (P < 0.05), IL-6 (P < 0.05) IL-8 (P < 0.05), and IL-10 (P < 0.05). The viral loads were positively correlated with IL-2 (P < 0.001), IL-5 (P < 0.05), IL-6 (P < 0.05) IL-8 (P < 0.05) and IL-10 (P < 0.05), while negatively correlated with CD 3 + T cells (P < 0.05) and CD3 + CD4 + T cells (P < 0.05). CONCLUSIONS: The viral loads are similar between asymptomatic, mild and severe or critical patients with COVID-19. The severity of COVID-19 may be related to underlying diseases such as diabetes mellitus. Lymphocyte subset and plasma cytokine levels may be as the markers to distinguish severely degrees of disease, and asymptomatic patients may be as an important source of infection for the COVID-19.

Treatment of coronavirus disease 2019 in Shandong, China: a cost and affordability analysis.

BACKGROUND: Coronavirus disease 2019 (COVID-19) is now a global public threat. Given the pandemic of COVID-19, the economic impact of COVID-19 is essential to add value to the policy-making process. We retrospectively conducted a cost and affordability analysis to determine the medical costs of COVID-19 patients in China, and also assess the factors affecting their costs. METHODS: This analysis was retrospectively conducted in Shandong Provincial Chest Hospital between 24 January and 16 March 2020. The total direct medical expenditures were analyzed by cost factors. We also assessed affordability by comparing the simulated out-of-pocket expenditure of COVID-19 cases relative to the per capita disposable income. Differences between groups were tested by student t test and Mann-Whitney test when appropriate. A multiple logistic regression model was built to determine the risk factors associated with high cost. RESULTS: A total of 70 COVID-19 patients were included in the analysis. The overall mean cost was USD 6827 per treated episode. The highest mean cost was observed in drug acquisition, accounting for 45.1% of the overall cost. Total mean cost was significantly higher in patients with pre-existing diseases compared to those without pre-existing diseases. Pre-existing diseases and the advanced disease severity were strongly associated with higher cost. Around USD 0.49 billion were expected for clinical manage of COVID-19 in China. Among rural households, the proportions of health insurance coverage should be increased to 70% for severe cases, and 80% for critically ill cases to avoid catastrophic health expenditure. CONCLUSIONS: Our data demonstrate that clinical management of COVID-19 patients incurs a great financial burden to national health insurance. The cost for drug acquisition is the major contributor to the medical cost, whereas the risk factors for higher cost are pre-existing diseases and severity of COVID-19. Improvement of insurance coverage will need to address the barriers of rural patients to avoid the occurrence of catastrophic health expenditure.

Killer cell immunoglobulin-like receptor genotypes and haplotypes with susceptibility to pulmonary tuberculosis infection.

BACKGROUND: Killer cell immunoglobulin-like receptors (KIRs) are expressed on natural killer (NK) cells and T cells and organized in highly polymorphic families. Genetic diversity is an important characteristic of KIR genes. The aim of the study was to investigate the influence of KIR genotypes and halotypes on the risk of pulmonary tuberculosis (PTB). METHODS: A sequence specific primer polymerase chain reaction (SSP-PCR) was employed to amplify the KIR genes and pseudogenes in 139 pulmonary tuberculosis (PTB) patients and 30 healthy controls. The innovative point of our study was the subdivision of the patient group according to sputum smear test (positive and negative). KIR genotype and haplotype frequencies were compared between the PTB group and the control group by Chi-square test, and p < 0.05 was regarded as statistically significant. RESULTS: The genotype AH and FZ14 may be associated with the clearance of Mycobacterium. In addition, haplotype B may be the susceptive haplotype that facilitated the clearance of Mycobacterium and haplotype A may be protective haplotype of PTB. CONCLUSIONS: Therefore, the diversity of genotypes and haplotypes induced an inflammatory reaction that resulted in continuous infection.

[A preliminary study on the relationship between HLA-Cw polymorphism and susceptibility to pulmonary tuberculosis].

OBJECTIVE: To analyze the relationship between HLA-Cw polymorphism and susceptibility to pulmonary tuberculosis (PTB), and therefore to explore the susceptible or resistant genes of PTB. METHODS: A hundred and twelve patients who were confirmed to have secondary PTB in Shandong Chest Hospital from May 2010 to May 2011 were selected as the PTB group, including 62 males and 50 females aged 19 - 69 years (mean 41.7). According to the acid-fast staining results, PTB patients were divided into a smear-negative group (SN group, 77 cases) and a smear-positive group (SP group, 35 cases). A hundred and ten subjects who underwent physical examination in Shandong Chest Hospital at the same period were selected as the control group, including 59 males and 51 females aged 21 - 67 years (mean 38.3). After genomic DNA was extracted, genotyping of HLA-Cw was conducted by sequence specific primer polymerase chain reaction (PCR-SSP) method. Then Hardy-Weinberg (H-W) equilibrium was tested, and gene frequencies(%) were estimated = 1-(1-phenotype frequencies)(1/2). Gene frequencies were compared between the PTB group and the control group, and between the SN group and SP group by χ(2) test. According to Bonferroni's principle, α was divided by the number of alleles (n = 8), and P < 0.006 25 was regarded as statistically significant. RESULTS: The frequency of HLA-Cw08 was significantly higher in PTB patients (43.6%, 75/112) compared with the controls (27.4%, 52/110), χ(2) = 8.790, P < 0.006 25. Among PTB patients, HLA-Cw04 had a significantly higher frequency in the SP group (20.7%, 13/35) than in the SN group (4.7%, 7/77), while HLA-Cw08 had a significantly lower frequency in the SP group (22.5%, 14/35) than in the SN group (54.4%, 61/77), χ(2) = 12.909, 16.732, both P < 0.006 25. CONCLUSIONS: HLA-Cw polymorphism is related to susceptibility to PTB. HLA-Cw08 may be one of the susceptible genes for PTB, and HLA-Cw04 and 08 may be related to MTB infectious status and clinical outcomes.

[Study on molecular characteristics regarding DNA genotype of Mycobacterium tuberculosis clinical strains in Shandong].

OBJECTIVE: To establish the molecular characteristics of Mycobacterium tuberculosis and on factors influencing the recent transmission of drug resistant isolates in Shandong. METHODS: Mycobacterium tuberculosis isolated from active pulmonary tuberculosis patients of 13 counties were genotyped by mycobacterial interspersed repetitive units (MIRU) methods. RESULTS: 12 loci of MIRU were detected in 558 isolates and a total of 143 MIRU patterns were confirmed. 66 isolates had distinct patterns, and 481 (86.2%) strains were in clusters. Shandong cluster included 177 strains with 74.6% of the isolates belonged to Beijing family. The recent transmission index of multi-drug resistance strains was in lower level, comparing to the susceptible strains. CONCLUSION: Our results showed that the Shandong cluster isolates had capacities of facilitating person-to-person transmission and high level of drug resistance.

[Application of mycobacterial interspersed repetitive units in typing Mycobacterium tuberculosis in Shandong].

OBJECTIVE: To evaluate the application of mycobacterial interspersed repetitive units (MIRU) in typing Mycobacterium tuberculosis in Shandong. METHOD: 826 strains of M. tuberculosis from laboratories of 12 county tuberculosis dispensaries in Shandong were cultured and typed by MIRU genotyping in the KICID laboratory. RESULTS: The 826 strains were typed to 201 distinct MIRU patterns, and 123 isolates were unique, 703 strains were grouped into 1 of 78 different MIRU clusters, and the MIRU genotype of the largest cluster was 223325173533. Among the clustered strains, 18 patients had an epidemiological history of contact. The Hunter-Gaston discriminatory index was 0.90. The allelic diversity of the sample was calculated for each other of the MIRU loci, and showed that MIRU26 had 10 alleles and was highly discriminative, while other five MIRU loci (MIRU31, 10, 39, 40, 4) were moderately discriminative. The reproducibility of the MIRU method was 100%. It took about RMB yen50 to genotype one strain. CONCLUSIONS: MIRU genotyping is a reproducible, fast, simple, and relatively cheap method. But because the isolates of 223325173533 genotype are predominant (30.89% of the isolates) in Shandong Province, it needs a second method IS6110 RFLP or by adding other more discriminative variable-number tandem repeat (VNTR) loci to improve the discriminative power.

[The short-term and long-term treatment outcomes in patients with pulmonary tuberculosis positive for drug-resistant and sensitive strains].

OBJECTIVES: To analyze the treatment outcomes in patients with smear positive tuberculosis, and to compare the difference in treatment response among patients infected with drug-sensitive and drug-resistant strains. METHODS: From 1998 to 2000, seven hundred and seventy-seven patients with primary smear-positive tuberculosis, which were from 30 surveillance sites, were followed for two years to monitor their treatment outcomes. RESULTS: At the completion of the 6 months' therapy, the overall rate of treatment failure was 1.8%, 2.6% for the drug-resistant cases and 1.6% for the drug-sensitive cases. Six-month follow-up showed a positive conversion rate of 2.7% in all the cases, 8.5% and 1.2% (P < 0.005) in the drug-resistant and the drug-sensitive cases respectively. One year follow-up showed that the positive conversion rate was 2.6% in all the cases, 6.9% and 1.6% (P < 0.005) in the drug-resistant and the drug-sensitive cases, respectively. Two-year follow-up showed an overall positive conversion rate of 1.3%, 1.0% and 1.3% in the drug-resistant and the drug-sensitive cases, respectively. Of the 152 drug-resistant cases, the rate of treatment failure was 2.6% at the completion of 6 months' therapy, but in cases with MDR-TB the rate was 10.3%. Six-month follow-up showed an overall positive conversion rate of 8.5%, but the rate reached 37.0% in cases with MDR-TB. One-year and two-year follow-up showed that the positive conversion rates were 6.9% and 1.0% respectively in all the drug-resistant cases, but 6.3% and 6.7% respectively in the MDR cases. CONCLUSIONS: Under the guidelines of the National Tuberculosis Program (NTP), the 2H(3)R(3)S(3)Z(3)/4H(3)R(3) regimen for primary smear-positive pulmonary TB was effective. But the cure rate was lower and the positive conversion rate higher in patients with MDR-TB.