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ABSTRACT BACKGROUND: Coronavirus disease 2019 (COVID-19) can cause cytokine release syndrome (CRS), which leads to high mortality rates. Tocilizumab suppresses CRS by blocking the signal transduction of interleukin-6 (IL-6). OBJECTIVE: To evaluate the clinical and laboratory parameters associated with mortality among patients receiving tocilizumab treatment. DESIGN AND SETTING: Retrospective observational study conducted in the chest disease departments of two different training and research hospitals in the center of Ankara, Turkey. METHODS: Patients who were hospitalized and treated with tocilizumab in September 2020 were retrospectively analyzed. Their laboratory parameters and clinical characteristics were obtained from the hospital information system database. Comparative analyses were performed between the patients who died and the ones who survived. RESULTS: A total of 58 patients who received tocilizumab treatment were included in this study, among whom 35 (60.3%) died. There was no difference between the mortality and survival groups in terms of white blood cell (WBC), neutrophil, lymphocyte, ferritin or C-reactive protein (CRP) levels detected on admission. WBC, lymphocyte, neutrophil and CRP levels measured on the third and fifth days after tocilizumab administration were found to be significantly lower in the survival group (P < 0.05). In multiple logistic regression analysis, age and oxygen saturation were determined to be independent risk factors for mortality. CONCLUSION: Persistently high WBC, CRP and neutrophil levels and low lymphocyte levels could be considered to be valuable indicators of mortality among COVID-19 patients treated with tocilizumab. Age and low oxygen saturation are independent risk factors for mortality among patients receiving tocilizumab treatment.
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@#SARS-CoV-2 has been spreading rapidly since its outbreak in December 2019.Understanding its epidemiological characteristics, especially cutting off transmission routes, is crucial to controlling the spread of the disease. In the study of transmission pathway, the issue of whether SARS-CoV-2 is transmitted through ocular surface tissue has also aroused concerns, but there are still no clinically confirmed cases and laboratory evidence of its infection through ocular surface tissue. New research suggests that the SARS-CoV-2 belongs to the same genus as SARS coronavirus(SARS-CoV), and that it enters cells in the same way as SARS-CoV. This paper reviews the research on SARS-CoV to investigate the possible mechanism of eye transmission of SARS-CoV-2.
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Diabetes mellitus (DM) has been shown to be an independent risk factor for developing the severe form of COVID-19, which in most cases requires management in intensive care units with mechanical ventilation and is associated with increased mortality. Objective Review and analyze the available scientific literature on COVID-19 in patients with Diabetes Mellitus Methods A search and analysis of the scientific literature was carried out in Pubmed, including publications in Spanish, English and French that deal directly with the topic diabetes mellitus and COVID-19. Results and Conclusions Adequate glycemic control has been shown to decrease mortality both in patients with previous diabetes and in those who develop hyperglycemia during hospitalization for COVID-19. For patients with diabetes mellitus that should be evaluated in an outpatient setting, telemedicine strategies are effective and should encompass nutritional management, adherence to treatment, and pharmacological aspects. Management with insulin therapy is the treatment of choice for hospitalized patients with moderate or severe COVID-19.
La diabetes mellitus (DM) ha demostrado ser un factor de riesgo independiente para desarrollar COVID-19 grave, la mayoría de estos casos requiere manejo en unidades de cuidado intensivo y se asocia a mayor mortalidad y costos sanitarios. Objetivo Realizar una búsqueda y análisis de la literatura científica disponible sobre COVID-19 en pacientes con Diabetes Mellitus. Métodos Se realizó una búsqueda sistemática y análisis de la literatura científica en Medline a través de PubMed, incluyendo publicaciones en español, inglés y francés que incluyan los siguientes términos de búsqueda: diabetes mellitus y COVID-19. Resultados y conclusiones Se ha demostrado que la hiperglucemia es un factor predictor para COVID-19 grave y se asocia a un incremento de mortalidad, así también se describe que un adecuado control glucémico disminuye la mortalidad tanto en pacientes con diabetes previa, como en aquellos que desarrollan hiperglucemia durante la hospitalización por COVID-19. El manejo con insulinoterapia es el tratamiento de elección para pacientes hospitalizados con COVID-19 moderada o severa. Para pacientes con diabetes mellitus que deben ser evaluados en un escenario ambulatorio, las estrategias de telemedicina son eficaces y deben abarcar el manejo nutricional, apego al tratamiento y aspectos farmacológicos
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Humans , Male , Female , Adult , Middle Aged , Aged , Therapeutics , Diabetes Mellitus, Type 2 , Insulin , SARS-CoV-2 , COVID-19ABSTRACT
Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.
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Severe Acute Respiratory Syndrome(SARS)is a deadly infectious disease caused by SARS Coronavirus(SARS-CoV).Inactivated SARS-CoV has been explored as a vaccine against SARS-CoV.However,safe and potent adjuvants,especially with more efficient and economical needle-free vaccination are always needed more urgently in a pandemic.The development of a safe and effective mucosal adjuvant and vaccine for prevention of emergent infectious diseases such as SARS will be an important advancement.PIKA,a stabilized derivative of Poly(I:C),was previously reported to be safe and potent as adjuvant in mouse models.In the present study,we demonstrated that the intraperitoneal and intranasal co-administration of inactivated SARS-CoV vaccine together with this improved Poly(I:C)derivative induced strong anti-SARS-CoV mucosal and systemic humoral immune responses with neutralizing activity against pseudotyped virus.Although intraperitoneal immunization of inactivated SARS-CoV vaccine alone could induce a certain level of neutralizing activity in serum as well as in mucosal sites,co-administration of inactivated SARS-CoV vaccine with PIKA as adjuvant could induce a much higher neutralizing activity.When intranasal immunization was used,PIKA was obligatorily for inducing neutralizing activity in serum as well as in mucosal sites and was correlated with both mucosal IgA and mucosal IgG response.Overall,PIKA could be a good mucosal adjuvant candidate for inactivated SARS-CoV vaccine for use in possible future pandemic.
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Objective To study the change of special antibodies titer IgG, IgM and nucleocaspid to SARS coronavirus (CoV) and observing the expression of stomach and enteric involvement on SARS-CoV infection by monoclonal antibody against N protein of SARS-CoV in the 7- year recovery period among family clustering cases of severe acute respiratory syndrome. Methods Special antibody titer to SARS-CoV of 14 patients from 5 different families and their 10 kinfolks continuously tested by IFA and antigen-capturing ELISA methods. Samples were taken in the 1st-7th year periods after SARS patients infected by SARS-CoV, being diluted and measured on it titers of three kinds of antibodies. Immunochemical staining with monoclonal antibody (mAb) against N protein of SARS-CoV was used to determine the stomach and enteric tissues among 5 SARS patients with their nucleocaspid antibody titer ascended obviously after 1st-7th year. Results When testing the IgG antibody titer of the 14 SARS patients by IFA method, the average titer was 1/71 (95%CI:1/58-1/85) in the 1st year, but began to descend in the following years, and the IgG antibody of the most SARS patients disappeared in the 7th year. Regarding the IgM titer, it disappeared in most of the SARS patients 1 year later. The average value of nucleocaspid antibody titer was 1/146 (95% CI:1/122-1/171) in the 1st year, and it descended as the IgG antibody titer did. In 5 cases, differences appeared.The nucleocaspid antibody titer was between 1/156 and 1/210 in 3 cases, and 2 cases were normal.Immunochemical staining with mAb against N protein of SARS-CoV was identified in the stomach and enteric tissues of 5 SARS patients with the nucleocaspid antibody titer increased significantly, 1st-7th year later. The five patients were detected by gastroscopy detection and cell immunohistochemistry test. 3 cases showed N protein antibody positive in the serum, and positive immunohistochemical expression in most of the cytoplasm in the gastric tissue mucous gland epithelial cells. 1 case also expressed in the intestinal tissue slurry columnar epithelium and interstitial cells. The other two cases showed negative on both serum N protein antibody and immunohistochemical expression. The biopsy results of the 5 patients were as follows: 1 case diagnosed as "signet-ring cell carcinoma of the stomach and rectum multiple transfer", 1 case of gastric polyp, 1 case of superficial antral gastritis and 2 cases were normal. Conclusion By testing the special IgG, IgM, nucleocaspid antibody to SARS-CoV of the 14 family clustering cases , we found that they all decreased in the 7th year, and most of them disappeared. The nucleocaspid antibody titer was related to pathogenetic condition. SARS-CoV was proved to be still present in stomach and enteric tissues of SARS patients with the nucleocaspid antibody titer increased significantly after the 7th year.
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A group of SARS-Iike coronaviruses(SL-CoV)have been identified in horseshoe bats.Despite SL-CoVs and SARS-CoV share identical genome structure and high-level sequence similarity,SL-CoV does not bind to the same cellular receptor as for SARS-CoV and the N-terminus of the S proteins only share 64% amino acid identity,suggesting there are fundamental differences between these two groups of coronaviruses.To gain insight into the basis of this difference,we established a recombinant adenovirus system expressing the S protein from SL-CoV(rAd-Rp3-S)to investigate its immune characterization.Our results showed that immunized mice generated strong humoral immune responses against the SL-CoV S protein.Moreover,a strong cellular immune response demonstrated by elevated IFN-γ and IL-6 levels was also observed in these mice.However,the induced antibody from these mice had weaker cross-reaction with the SARS-CoV S protein,and did not neutralize HIV pseudotyped with SARS-CoV S protein.These results demonstrated that the immunogenicity of the SL-CoV S protein is distinct from that of SARS-CoV,which may cause the immunological differences between human SARS-CoV and bat SL-CoV.Furthermore,the recombinant virus could serve as a potential vaccine candidate against bat SL-CoV infection.
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To evaluate the immunogenicity of inactivated SARS coronavirus (SARS-CoV), three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant, adjuvant,and normal saline respectively. Eight batchs of serum were sampled from the auricular vein at day 7 to day 51, and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test. Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining. The results showed that, rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity, which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses. The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively. In the experimental group, no obvious histopathological changes was detected in the H&E stained slides of heart, spleen, kidney and testis samples, but the livers had slight histopathological changes, and the lungs presented remarkable histopathological changes. These findings are of importance for SARS-CoV inactivated vaccine development.
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Objective To develop plaque assay for SARS virus, in order to provide a reliable means for SARS research. Methods SARS virus BJ 01 strain in various dilutions was inoculated to Vero E6 cells, the cells were then covered with nutritious agar. The titers of the plaque were measured by Dullbecco R method. Results The plaque of SARS virus appeared on the third day after the cell cultures infected with virus. The plaque was round in shape, 2.5-3 mm in diameter. Conclusion The plaque assay developed in present study was stable and regular, and it could be used in SARS research.
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SARS coronavims is an emerging virus. A lot of animals could be infected by SARS-CoV and Himalayan palm civets, as one of important hosts, is an ideal animal model. Viral genetic factors have been implicated in the emergence of SARS-CoV, with the suggestion that this virus is a recombinant between mammalian and avian coronaviruses. However, the recombination is unlikely to explain the appearance of SARS in humans.
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Objective:To obtain the spike glycoprotein(S protein) of SARS coronavirus (SARS-Cov) for detection of the corresponding antibody and preparation of diagnostic kits.Methods:Genes encoding S protein were picked out to form a new gene and divided into two parts according to the prediction of antigenicty.They were obtained by nucleotide synthesis and were cloned into expression vector pET32a(+) respectively.Proteins then were induced by IPTG and purified through Ni-NTA.Results:Target proteins were successfully expressed and purified.Conclusion:The recombinant S proteins of SARS-CoV could be of great help in diagnosing the disease and investigating the virus further.
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Objective:To prepare monoclonal antibodies with high specificity against different regions of N protein and to analyze antigenic epitopes of SARS-CoV N protein.Methods: Balb/ c mice were immunized with purified different region N protein to prepare McAb by hybridoma technique and select by indirect ELISA.Both of these McAbs were characterized for their antibody titre in the culture supernatant as well as ascites,class,subclass and affinity analysis.The antigen specificity for McAb were evaluated and confirmed by Western blot.The binding site and capability of McAb were observed by ELISA additive assay.Results: Seven hybridoma cell lines secreting monoclonal antibodies against SARS-CoV N1 and two hybridoma cell lines secreting monoclonal antibodies against SARS-CoV N2 were obtained.About the immunoglobulin subclass of McAb,six were IgG1,two were IgG2b and one was IgG3.Western blot showed that the McAbs reacted specifically with SARS-CoV N protein.ELISA additive assay indicated that two antibodies against SARS-CoV N1 recognized same epitopes,while others recognized distinct epitopes.Conclusion: We successfully obtained nine specific McAbs against the different region SARS-CoV N protein after expression and purification of the different part SARS-CoV N protein.It shows that the N1 McAb recognizs the N-terminus of the N protein whereas the N2 McAb recognizs the C-terminus.
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Objective:To evaluate the immunogenicity of an experimental SARS-CoV inactivated vaccine.Methods:The virus suspension of F69 strain was inactivated with 0.4% formaldehyde and purified,then used as the immune antigen combined with Freund′s adjuvant.Eight adult New Zealand rabbits were immunized 4 times with this vaccine.12 sets of rabbit serum were sampled from the third day to 74th day after first immunization.Titers of specific IgG antibody and neutralizing antibody were determined by indirect ELISA and micro-cytopathic effect neutralizing test.Results:Rapid and potent humoral immune responses were induced by F69 inactivated vaccine in all eight immunized rabbits.Both specific IgG antibody and neutralizing antibody all peaked just with 2 vaccinations,with the maximum titer of 1∶81 920 and 1∶20 480 respectively about 6 weeks after first immunization.Across neutralizing reaction existed between F69 and Z2-Y3 strains.Conclusion:F69 inactivated vaccine owns strong immunogenicity.Similar antigenic epitopes exist between the F69 strain and Z2-Y3 strain,which ensured the cross neutralizing reaction.
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Objective To predict the B cell epitopes for S protein of severe acute respiratory syndrome (SARS) coronavirus. Methods Based on SARS coronavirus genome sequence and the analysis of the flexible regions of secondary structure for S protein, the B cell epitopes for S protein were predicted by methods of Kyte Doolittle hydrophilicity plot, Emini, and Jameson Wolf. Results The computer predicted most possible epitopes for S protein were located within or nearby its N terminal No. 91-98, 1121-1153 and 185-193. The N terminal No. 1050-1059, 752-765, 41-50, 666-674, 264-287, 340-348, 408-416, and 424-439 may be the possible B cell epitopes. Conclusion Prediction of the B cell epitopes for the S protein based on multiple parameters is helpful for the identification of B cell epitopes using experimental methods.
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Objective To explore the effects of severe acute respiratory syndrome coronavirus (SARS-CoV) on heart and its conduction system in SARS patients. Methods Six specimens of heart tissue and one specimen of heart condunction system from patients who died from SARS were studied histologically, and by histochemical and in situ hybridization examinations. Results The pathological changes showed that a part of cardiomyocytes manifested slight vacuolar degeneration, atrophy and cytoplasmic lysis, stromal edema, mild mononuclear infiltration, and vasculitis. SARS-CoV was identified within some cardiomyocytes and specialized cardiomyocytes which belonged to the conduction system of the heart by in situ hybridization in combination with Macchiavello's viral inclusion stain. Conclusions The results showed that SARS-CoV could invade not only cardiomyocytes, but also the specialized cells of heart conduction system, thus resulting in mild viral myocarditis-like pathological changes. The results provided the evidence in explaining the clinical manifestations of cardiac dysfunction in patients with SARS.