Ubiquitin-specific protease 8 inhibits lipopolysaccharide-triggered pyroptosis of human bronchial epithelial cells by regulating PI3K/AKT and NF-kB pathways

Asthma, characterized by dysfunction of airway epithelial cells, is regarded as a chronic inflammatory disorder in the airway. Ubiquitin-specific protease 8 (USP8) belongs to ubiquitin proteasome system and mediates the stability of E3 ligases. The anti-inflammatory effect of USP8 has been widely investigated in distinct diseases, while the role of USP8 in asthma remains elusive. Firstly, human bronchial epithelial cells (BEAS-2B) were treated with lipopolysaccharide, which reduced the cell viability of BEAS-2B and induced the secretion of lactate dehydrogenase (LDH). Moreover, the expression of USP8 was downregulated in BEAS-2B post lipopolysaccharide treatment. Secondly, overexpression of USP8 enhanced cell viability of lipopolysaccharide-treated BEAS-2B, and reduced the LDH secretion. USP8 overexpression also attenuated lipopolysaccharide-induced upregulation of TNF-α, IL-6, and IL-1β in BEAS-2B. Thirdly, lipopolysaccharide treatment promoted the expression of NLRP3 (NLR Family Pyrin Domain Containing 3), N-terminal domain of gasdermin D (GSDMD-N), caspase-1, IL-1β, and IL-18 in BEAS-2B, which was inhibited by USP8 overexpression. Lastly, USP8 overexpression decreased the phosphorylation of NF-κB, while it increased the phosphorylation of PI3K and AKT in lipopolysaccharide-treated BEAS-2B. In conclusion, USP8 inhibited lipopolysaccharide-triggered inflammation and pyroptosis in human bronchial epithelial cells by activating PI3K/AKT signaling and inhibiting NF-κB signaling pathway (AU)

Risk factors for hospitalization of pregnant women with influenza A / 中华传染病杂志

Objective:To investigate the influencing factors of hospitalization for pregnant women with influenza A.Methods:From December 2018 to February 2019, 261 pregnant women with influenza A were admitted to Beijing Ditan Hospital, Capital Medical University. The clinical data of age, gestational period, underlying diseases, time from onset to treatment, white blood cell count and lymphocyte count of these patients were collected. Data of out-patients were compared with those of inpatients. Chi-square test and multivariate logistic regression were used to analyze the influencing factors of hospitalization in pregnant women with influenza A.Results:Among the 261 cases of pregnancy with influenza A, 36 cases (13.79%) were hospitalized, of which 10 (27.78%) were hospitalized due to severe influenza complications, the other 26 cases (72.22%) were hospitalized due to pregnancy related adverse events. The proportions of hospitalized patients with age ≥30 years old, gestational period ≥28 weeks, combined with underlying diseases and lymphocyte count <1×10 9/L were 75.00%(27/36), 83.33%(30/36), 16.67%(6/36) and 50.00%(18/36), respectively, which were significantly higher than those of out-patients (47.11%(106/225), 35.56%(80/225), 0.89%(2/225) and 13.22%(16/121), respectively; χ2=9.66, 29.05, 26.00 and 22.12, respectively, all P<0.05). The proportions of inpatients and out-patients with white blood cell count ≥4×10 9/L were 97.22%(35/36) and 97.52%(118/121), respectively, and there was no significant difference ( χ2=0.01, P=0.921). Multivariate logistic regression analysis showed that age ≥30 years (odds ratio ( OR)=5.181, 95% confidence interval ( CI) 1.628-16.489, P=0.005), gestational period ≥28 weeks ( OR=11.054, 95% CI 3.233-37.796, P<0.01), lymphocyte count <1×10 9/L ( OR=6.864, 95% CI 2.237-20.729, P=0.001), and time from onset to treatment <24 h ( OR=0.076, 95% CI 0.012-0.468, P=0.005) were the influencing factors for hospitalization of pregnant women with influenza A. Conclusion:Age ≥30 years old, gestational period ≥28 weeks, lymphocyte count <1×10 9/L and time from onset to treatment <24 h are the influencing factors for hospitalization of pregnant women with influenza A.

Epidemiological and clinical characteristics of 69 cases with imported corona virus disease 2019 / 中华传染病杂志

Objective:To analyze the epidemiological and clinical characteristics of patients with imported corona virus disease 2019 (COVID-19) in Beijing City.Methods:A case-control study was performed to retrospectively analyze 69 cases of imported COVID-19 from abroad and 147 cases of domestic confirmed COVID-19 from China as a control group from January 20 to March 20, 2020 admitted to Beijing Ditan Hospital, Capital Medical University.The epidemiological and clinical characteristics were compared.Statistical analysis were performed by t test, Mann-Whitney U test, chi-square test and Fisher exact test. Results:The main sources of the cases in the import group were from the United Kingdom, Italy, Spain and other European countries, with 44.9%(31/69) of the overseas students entering the country by air. The age of the imported group (27(21, 40) years) was lower than the domestic group (43 (32, 59)years), the difference between the two groups was statistically significant ( U=2 828.500, P<0.01). Compared with the domestic group, the proportion of cases with contact history of confirmed cases in the imported group was lower (30.4%(21/69) vs 68.0%(100/147)), the interval between onset and admission ≤seven days was higher (81.2%(56/69) vs 66.0%(97/147)), the proportion of cases with underlying diseases was lower (21.7%(15/69) vs 44.2%(65/147)). The differences between the two groups were all statistically significant ( χ2=26.935, 5.233 and 10.175, respectively, all P<0.05). The proportion of mild cases in the imported group was higher than that in the domestic group (42.0%(29/69) vs 10.9%(16/147)). Seventeen cases with olfactory abnormality and 12 cases with taste abnormality were found in the imported group, while no olfactory and taste abnormality was found in the domestic group. The proportions of fever, weakness, muscle soreness and dyspnea were all lower than those of the domestic group, the differences between the two groups were all statistically significant ( χ2=13.851, 8.118, 9.730 and 16.255, respectively, all P<0.01). The proportions of cases with decreased lymphocyte absolute numbers (37.7%(26/69) vs 67.3%(99/147)) and increased C reactive protein level (15.9%(11/69) vs 51.8%(72/139)) were both lower than the domestic group, and the differences between the two groups were both statistically significant ( χ2=18.015 and 24.722, respectively, both P<0.01). The proportions of cases with ground glass shadow and consolidation of chest computed tomography were lower than those of the domestic group and the differences between the two groups were all statistically significant ( χ2=11.961 and 5.099, respectively, all P<0.05). In terms of complications, the proportions of cases with acute respiratory distress syndrome and acute myocardial injury were lower (2.9%(2/69) vs 10.9%(16/147) and 4.3%(3/69) vs 14.0%(16/114), respectively), and there were statistically significant differences between the two groups ( χ2=4.017 and 4.335, respectively, both P<0.05). There were no cases received mechanical ventilation and extracorporeal membrane oxygenation in the imported group, and the proportions of patients received oxygen therapy and antibiotic treatment were significantly lower than those in the domestic group (13.0%(9/69) vs 26.5%(39/147) and 13.0%(9/69) vs 39.5%(58/147), respectively) and the differences between the two groups were statistically significant ( χ2=4.942 and 15.797, respectively, both P<0.05). Conclusions:The majority of imported COVID-19 cases are mainly from European countries, mostly young and middle-aged, and mostly mild and ordinary types.The symptoms of olfactory and taste abnormality are found for the first time.

Interpretation for diagnosis of Yersiniosis / 中华流行病学杂志

Yersiniosis is one of the "other infectious diarrhea" of the notifiable infectious diseases and also an important food-borne disease. However, it lacked the basis or standard for diagnosis. The Chinese Preventive Medicine Association coordinated experienced researchers from National Institute for Communicable Disease Control and Prevention, China CDC and other institutes to produce the group standard entitled "Diagnosis of Yersiniosis" (T/CPMA 005-2019). Based on the principle of "legality, scientificity, advancement, and feasibility" , the standard gives a clear definition for Yerisiniosis, stipulates diagnosis basis, principles and main differential diagnosis and provides two informative appendixes for epidemiological and clinical characteristics and a normative appendix for laboratory detection. The standard provides accurate basis and methods of Yersiniosis diagnosis for hospitals and CDCs at all levels in China. It will solve the problems that Yersiniosis cannot be clearly diagnosed for clinical cases and in the outbreaks.

Diagnostic sensitivity and related impact factors of T cell spot test of tuberculosis in acquired immunodeficiency syndrome/tuberculosis patients / 中华传染病杂志

Objective To analyze the diagnostic sensitivity and related impact factors of T cell spot test of tuberculosis (T‐SPOT .TB ) test in acquired immunodeficiency syndrome (AIDS )/tuberculosis (TB) patients .Methods Ninety‐two confirmed cases with AIDS/TB coinfection were tested by T‐SPOT . TB ,and the impact of CD4+ T cells counts on the diagnostic sensitivity was analyzed .Multivariate Logistic analysis was used for the analysis of impact factors of T‐SPOT .TB sensitivity .Blood samples of 19 cases with advanced stage AIDS/TB from January 2015 to January 2016 were collected ,and peripheral blood mononuclear cell (PBMC ) were isolated by Ficoll method , and lymphocytes were isolated by Percoll method .McNemar test was used for the comparison of these two methods .Results Among the 92 patients with AIDS/TB ,T‐SPOT .TB tests were positive in 51 cases ,with positive rate of 55 .4% .The sensitivity was 26 .3% (10/38) when CD4+ T cell count less than 20/μL ,and that was 92 .9% (13/14) when CD4+ T cell more than 200/μL .In Logistic analysis ,the sensitivity of T‐SPOT .TB test in patients with extra‐pulmonary tuberculosis was better than that in pulmonary tuberculosis patients (OR=3 .042 , P=0 .038) .The sensitivity of T‐SPOT .TB was positively correlated with CD4+ T cell count ,and the sensitivity increased by 2 .889 times when CD4+ T cells increasing 100/μL (OR=3 .889 ,P=0 .016) .The percentage of lymphocytes in PBMC was also positively correlated with T‐SPOT .TB positivity ,and the sensitivity increased by 1 .393 times when the percentage increasing 30% (OR=2 .393 ,P=0 .045) .When Percoll was used for lymphocytes isolation ,the T‐SPOT .TB sensitivity was 52 .6% (10/19) ,and when Ficoll was used for PBMC isolation ,the sensitivity was 36 .8% (7/19) .The difference was not statistically significant (P=0 .375) .Conclusion The sensitivity of T‐SPOT .TB test based on lymphocytes is higher than that based on PBMC .

Comparison analysis of clinical features of varicella in pregnant women and outcomes of mothers and their newborns / 中华传染病杂志

Objective To investigate the clinical features of varicella and the prognosis in pregnant women and their newborns .Methods Clinical data of pregnant women with varicella zoster virus (VZV) infection (n= 25) and their newborns hospitalized in Beijing Ditan Hospital from 1st Jan .2008 to 31st Dec .2014 were retrospectively analyzed and randomly compared to non‐pregnant women with VZV infection (n=50) .Clinical features and prognosis of varicella in pregnant women and their infants were analyzed .Chi‐square test was used for categorical data and t test was used for quantitative data .Results Time to rash scab of varicella in pregnant women was longer than non‐pregnant women ([10 .1 ± 2 .1] d vs [5 .6 ± 1 .4] d ,t=10 .941 ,P<0 .05) .The rate of bacterial infection in pregnant women was higher than non‐pregnant varicella women (72 .0% [18/25] vs 32 .0% [16/50] ,χ2 = 10 .761 , P < 0 .05) , with statistical significance .Among 25 cases of varicella pregnant women ,the pregnancy complications were observed in 3 cases of diabetes ,2 cases of premature rupture of membranes ,5 cases of anemia and 1 case of oligoamnios .Seven cases out of 25 pregnant women underwent parturition during fever and varicella period ,and 3 cases (12 .0% ) were complicated with intrapartum hemorrhage . Twenty five varicella pregnant women were all cured after antiviral and supportive treatment and gave birth to their babies ,with no abortion ,stillbirth or birth defects .No congenital varicella was observed in newborns .Of the 25 infants ,4 developed (16 .0% ) varicella within 2 weeks after birth and they were all born from mothers who developed varicella around delivery time .The clinical features of neonatal varicella presented with classic rash with no fever .The time to rash scab was longer (11 .0 ± 2 .1) d and antibody test for VZV was negative .All neonates were cured after antiviral and immunoglobulin treatment .Conclusions Longer duration of skin rash scab and higher rate of bacterial infections are the features of varicella in pregnant women .Intrapartum hemorrhage occurrs more commonly in pregnant women with varicella onset around delivery time .Varicella occurring during mid‐pregnancy may not increase the risk of neonatal birth defects after treatment .The newborns whose mothers with varicella onset during perinatal time especially around delivery time may suffer from varicella .The prognosis of neonatal period varicella is good after treatment .

Clinical analysis of early hemolysis and delayed hemolysis after treatment in patients with malaria / 中华传染病杂志

Objective To investigate the risk factors of delayed hemolysis after treatment in patients with malaria .Methods Eighty-nine cases of malaria were retrospectively analyzed .The incidence rate , time from treatment to delayed hemolysis and clinical features of delayed hemolysis after treatment in patients with malaria were investigated .The characteristics of demography ,etiology and laboratory data were compared between delayed hemolysis group and non-delayed hemolysis group .The t test ,χ2 test and Fisher exact test were used for comparison between groups .Results A total of 89 cases of malaria infection were included and 8 cases were diagnosed with delayed hemolysis after treatment among them , with incidence rate of 8 .99% .Patients developed delayed hemolysis after anti-malarial treatment with a median of 7 .5 d and patients recovered from hemolysis after the usage of glucocorticoid with a median of 2 .5 d .The 8 cases were all infected with Plasmodium f alciparum ,and 4 of which had high parasitemia . None of the patients with delayed hemolysis came from epidemic area ,while 28 of the patients without non-delayed hemolysis came from epidemic area .The difference was statistically significant (P=0 .042 , Fisher unilateral exact test) .The average level of minimum hemoglobin was (44 .87 ± 11 .58) g/L in patients with delayed hemolysis ,which was significantly lower than that of non-delayed hemolysis group (108 .35 ± 19 .72) g/L (t= -8 .923 , P< 0 .01) .Conclusion Plasmodium falciparum infection , hyperparasitemia and having no immunity against malaria may be risk factors of delayed hemolysis after treatment .

Clinical study on peripheral T lymphocyte subsets in children with hand-foot-and-mouth disease / 中华传染病杂志

Objective To evaluate the dynamic changes of T lymphocyte subsets in children with hand-foot-and-mouth disease(HFMD)and to provide new evidence for the therapy and prognosis.Methods Peripheral venous blood samples of 346 HFMD cases in acute stage who were hospitalized in Beijing Ditan Hospital from May 1,2008 to August 31,2008 were collected and T lymphocyte subsets were assayed by flow cytometer.Meanwhile,T lymphocyte subsets of 67 HFMD cases in recovery phase were also detected.The pathogens were determined by reverse transcriptionpolymerase chain reaction(RT-PCR)using pharynx swab samples from 99 cases.Different samples were compared by independent-sample t test,paired t test or variance analysis.Results The average levels of T lymphocyte subsets of HFMD children in different agc groups were all lower than reference levels of healthy children in according age groups.In severe cases.T lymphocyte(TL)/lymphocyte (L)ratio in all age groups,helper T cell(Th)/L ratio in children older than 1 year,TL,Th and Th/suppressor T cell(Ts)ratio in children of 1-2 years old were all lower than those in common eases (P＜0.05).The Th/L ratio tended to increase with the disease progression.Ratios of TL/L and Th/L in common cases were increased in recovery phase(TL/L:56.3±8.6 vs 61.1±9.1,t=2.56,P＜0.05;Th/L:30.2±7.2 vs 34.9±7.9,t=2.90,P＜0.05)and all indices of severe cases except Ts/Lratio and Th/Ts ratio increased apparently in recovery phase(P＜0.01).TL[(1.738±0.976)×10~6/Lvs(2.696±1.946)×10~6/L,t=2.17,P＜0.05],Th/L ratio(25.9±7.0 vs 30.2±7.2,t=2.34,P＜0.05),Th[(0.864±0.550)×10~6/L vs(1.459±0.879)×10~6/L,t=2.90,P＜0.01]and L[(3.352±1.458)×10~6/L vs(4.664±2.435)×10~6/L,t=2.32,P＜0.05]of severe cases in acute phase were all lower than those of common cases(P＜0.05),while those were not significantly different in recovery phase between two groups(P＞0.05).The T lymphocyte subsets of enterovirus(EV)71 positive cases were lower than EVT1 negative cases,but there was no significant difference between these two groups(P＞0.05).Conclusion T lymphocyte immune responses may be correlated with HFMD onset and progression.

Clinical study of 15 children with hand foot and month disease and acute flaccid paralysis / 中华急诊医学杂志

Objective To discuss the clinical characteristics and prognosis of 15 children with hand foot and mouth disease (HFMD) and acute flaccid paralysis (AFP) who were admitted to Beijing Ditan Hospital during the outbreak of HFMD in 2008. Method The epidemiology, clinical manifestations, cerebrospinal fluid (CSF),magnetic resonance imaging and prognosis of 15 children with HFMD and AFP were retrospectively reviewed. The recovery of the patients' affected extremities were monitored for 4 weeks. Results The mean age of these patients was (22.47 ± 20.68) months (range: 5～72 months). Acute paralysis developed (3.47 ± 1.68) days after the onset of fever and progressed to maximum severity within (1～2) days. Poliomyelitis-like syndrome was observed in all cases. Of the 15 cases, 10 had monoplegia of lower limbs, two had paraplegia, one had monoplegia of upper limbs and two had quadriplegia. In these cases, the muscle power varied from level 0 to level 4, and six even showed no muscle power in their affected extremities. Thirteen cases developed neurologic complications (encephalitis, meningitis or ataxia) and three had transient urinary retention. Cerebrospinal MRI examination in eight cases showed hyperintense lesions on T2-weighted images, predominantly in the impaired anterior horn regions of the spinal cord (C2～C7 for cases with upper extremity impairments and T12～L1 for cases with lower extremity impairments), and displayed long T1 signals and long T2 signals. In addition, the midbrain, brain-stem or medulla was also involved in four cases who also contracted encephalitis or meningitis. The muscle strength in 11 patients with single lower extremity impairment showed improvements in the distal limb muscles within 4～8 days, and the other cases showed recovery 2～3 weeks later. Conclusions HFMD in combination with AFP most commonly occurs in children aged less than 2 years old. Acute paralysis develops during the early stage of infection and progresses to a maximum severity within 2 days. In most cases described here, paralysis occurred in a single lower extremity and recovered more rapidly than those with all four limbs affected or with single upper extremity impairment . MRI examination is particularly valuable for the diagnosis and prognosis of AFP because of its high sensitivity and accuracy.

A clinical analysis of 33 cases of H1N1 influenza A / 中华内科杂志

Objective To review the clinical characteristics of H1N1 influenza A, and suggest the clinical practices for the diagnosis and treatment of H1N1 influenza A in the future. Methods Thirty-three cases of H1N1 influenza A hospitalized in Beijing Ditan Hospital from May 15 to June 22, 2009 were studied and the clinical data were statistically analyzed with SPSS 11.0 for Windows. Results Twenty-five of the 33 patients had a history of travelling in America, Canada, Japan etc within a week; the latent period was between 1 and 6 days in 12 close contact patients. The main symptoms of H1N1 influenza A are fever (66.7%), dry cough (60.6%), cough with sputum (42.4%) and sore throat (36.4%). The laboratory tests in 24 cases(72.7%) were normal, while mild abnormal results were found in the remaining patients.All of the 33 cases were discharged according to the Standard of Diagnosis and Treatment of H1N1 Influenza A published by The Minister of Health, China. The period between 2-consecutive negative results in viral nucleic acid RT-PCR detection and the presenting symptom was 2 to 16 days and the period of hospitalization was 3 to 16 days. Conclusion The new type of H1N1 influenza A is characterised by mild symptoms, short period of hospitalization and good prognosis. All the patients can be cured, if they do not suffer from other severe chronic disease.

Reduced sera neutralization to Omicron SARS-CoV-2 by bothinactivated and protein subunit vaccines and the convalescents

Omicron variant continues to spread all over the world. There are lots of scientific questions remaining to be answered for such a devastating variant. There are a dozen of vaccines already in clinical use. The very urgent scientific question would be whether or not these vaccines can protect Omicron variant. Here, we tested the sera from both convalescents and vaccine recipients receiving either inactivated or protein subunits vaccines (CoronaVac from Sinovac, or BBIBP-CoV from Sinopharm, or ZF2001 from Zhifei longcom) for the binding antibody titers (ELISA) and neutralization antibodies titers (pseudovirus neutralization assay). We showed that Omicron do have severe immune escape in convalescents, with 15 of 16 were negative in neutralization. By contrast, in vaccinees who received three jabs of inactivated or protein subunit vaccine, the neutralizing activity was much better preserved. Especially in the ZF2001 group with an extended period of the second and third jab (4-6 months) remains 100% positive in Omicron neutralization, with only 3.1-folds reduction in neutralizing antibody (NAb) titer. In this case, we proposed that, the multi-boost strategy with an extended interval between the second and third jab for immune maturation would be beneficial for NAb against devastating variants such as Omicron.

Specific Dynamic Variations in the Peripheral Blood Lymphocyte Subsets in COVID-19 and Severe Influenza A Patients: A Retrospective Observational Study

BackgroundBoth COVID-19 and influenza A contribute to increased mortality among the elderly and those with existing comorbidities. Changes in the underlying immune mechanisms determine patient prognosis. This study aimed to analyze the role of lymphocyte subsets in the immunopathogenesisof COVID-19 and severe influenza A, and examined the clinical significance of their alterations in the prognosis and recovery duration. MethodsBy retrospectively reviewing of patients in four groups (healthy controls, severe influenza A, non-severe COVID-19 and severe COVID-19) who were admitted to Ditan hospital between 2018 to 2020, we performed flow cytometric analysis and compared the absolute counts of leukocytes, lymphocytes, and lymphocyte subsets of the patients at different time points (weeks 1- 4). ResultsWe reviewed the patients data of 110 healthy blood donors, 80 Non-severe-COVID-19, 19 Severe-COVID-19 and 43 severe influenza A. We found total lymphocytes (0.93 x109/L, 0.84 x109/L vs 1.78 x109/L, P < 0.0001) and lymphocyte subsets (T cells, CD4+ and CD8+ T cell subsets) of both severe patients to be significantly lower than those of healthy donors at early infection stages. Further, significant dynamic variations were observed at different time points (weeks 1-4). ConclusionsOur study indicates lymphopenia to be associated with disease severity and suggests the plausible role of lymphocyte subsets in disease progression, which in turn affects prognosis and recovery duration in patients with severe COVID-19 and influenza A.

Comprehensive genomic, immunological and clinical analysis of COVID-19 vaccine breakthrough infections: a prospective, comparative cohort study

ObjectivesAs the COVID-19 pandemic is still ongoing and SARS-CoV-2 variants are circulating worldwide, an increasing number of breakthrough infections have been detected despite the good efficacy of COVID-19 vaccines. MethodsA prospective, comparative cohort study was conducted in Beijing Ditan Hospital to evaluate the clinical, immunological and genomic characteristics of COVID-19 breakthrough infections. Data on 88 COVID-19 breakthrough cases (vaccinated group) and 41 unvaccinated cases (unvaccinated group) from June 1 to August 20, 2021 were extracted from a cloud database. Among these 129 COVID-19 cases, we successfully sequenced 33 whole genomes, including 16 from the vaccinated group and 17 from the unvaccinated group. ResultsAsymptomatic and mild cases predominated in both groups, but 2 patients developed severe disease in the unvaccinated group. Between the two groups, the median time of viral shedding in the vaccinated group were significantly lower than those in the unvaccinated group (p = 0.003). A comparison of dynamic IgG titres of cases in the two groups indicated that IgG titres in the vaccinated group showed a significantly increasing trend (P =0.028). The CD4+T lymphocyte count was lower in the unvaccinated group, and there was a significant difference between the two groups (p=0.018). In the vaccinated group, the number of moderate cases who received Sinopharm BBIBP (42 cases) was significantly higher than those who received Sinovac Coronavac (p=0.020). Whole-genome sequencing revealed 23 cases of delta variants, including 15 patients from the vaccinated group. However, no significant difference was observed in either the RT-qPCR results or viral shedding time. ConclusionsCOVID-19 vaccine breakthrough infections were mainly asymptomatic and mild, the IgG titres were significantly higher and increased rapidly, and the viral shedding was short. Delta variants may be more likely to cause breakthrough infections, and vaccination may not reduce the viral loads and shedding time.

Impact of various vaccine boosters on neutralization against Omicron following prime vaccinations with inactivated or adenovirus-vectored vaccine

Since the first report on November 24, 2021, the Omicron SARS-CoV-2 variant is now overwhelmingly spreading across the world. Two SARS-CoV-2 inactivated vaccines (IAVs), one recombinant protein subunit vaccine (PRV), and one adenovirus-vectored vaccine (AdV) have been widely administrated in many countries including China to pursue herd immunity. Here we investigated cross-neutralizing activities in 341 human serum specimens elicited by full-course vaccinations with IAV, PRV and AdV, and by various vaccine boosters following prime IAV and AdV vaccinations. We found that all types of vaccines induced significantly lower neutralizing antibody titers against the Omicron variant than against the prototype strain. For prime vaccinations with IAV and AdV, heterologous boosters with AdV and PRV, respectively, elevated serum Omicron-neutralizing activities to the highest degrees. In a mouse model, we further demonstrated that among a series of variant-derived RBD-encoding mRNA vaccine boosters, it is only the Omicron booster that significantly enhanced Omicron neutralizing antibody titers compared with the prototype booster following a prime immunization with a prototype S-encoding mRNA vaccine candidate. In summary, our systematical investigations of various vaccine boosters inform potential booster administrations in the future to combat the Omicron variant.

Safety and immunogenicity of a recombinant tandem-repeat dimeric RBD protein vaccine against COVID-19 in adults: pooled analysis of two randomized, double-blind, placebo-controlled, phase 1 and 2 trials

BackgroundA safe and effective coronavirus disease 2019 (COVID-19) vaccine is urgently needed to control the ongoing pandemic. Although progress has been made recently with several candidates reporting positive efficacy results, COVID-19 vaccines developed so far cannot meet the global vaccine demand. We developed a protein subunit vaccine against COVID-19, using dimeric form of receptor-binding domain (RBD) as the antigen. We aimed to assess the safety and immunogenicity of this vaccine in humans and determine the appropriate dose and schedule for an efficacy study. MethodsWe did two randomized, double-blind, placebo-controlled, phase 1 and 2 trials for an RBD-based protein subunit vaccine, ZF2001. In phase 1 study, 50 healthy adults aged 18-59 years were enrolled and randomly allocated to three groups to receive three doses of vaccine (25 g or 50 g RBD-dimer, with adjuvant) or placebo (adjuvant-only) intramuscularly, 30 days apart. In phase 2 study, 900 healthy adults aged 18-59 years were enrolled and randomly allocated to six groups to receive vaccine (25 g or 50 g RBD-dimer, with adjuvant) or placebo (adjuvant-only) intramuscularly, with the former 3 groups given two doses and the latter 3 groups given three doses, 30 days apart. For phase 1 trial, the primary outcome was safety, as measured by the occurrence of adverse events and serious adverse events. The secondary outcome was immunogenicity as measured by the seroconversion rate and magnitude of antigen-binding antibodies, neutralizing antibodies and T-cell cytokine production. For phase 2 trial, the primary outcome included both safety and immunogenicity. These trials are registered with ClinicaTrials.gov, NCT04445194 and NCT04466085. FindingsBetween June 22 and September 15, 2020, 50 participants were enrolled to the phase 1 study (mean age 32.6 years) and 900 participants were enrolled to phase 2 study (mean age 43.5 years), to receive vaccine or placebo with a two-dose or three-dose schedule. For both trials, local and systemic adverse reactions were absent or mild in most participants. There were no serious adverse events related to vaccine in either trial. After three doses, neutralizing antibodies were detected in all participants receiving either 25 g or 50 g dose of vaccine in phase 1 study, and in 97% (the 25 g group) and 93% (the 50 g group) of participants, respectively, in phase 2 study. The SARS-CoV-2-neutralizing geometric mean titres (GMTs) were 94.5 for the 25 g group and 117.8 for the 50 g group in phase 1, and 102.5 for the 25 g group and 69.1 for the 50 g group in phase 2, exceeding the level of a panel of COVID-19 convalescent samples (GMT, 51). Vaccine induced balanced TH1 and TH2 responses. The 50 g group did not show enhanced immunogenicity compared with the 25 g group. InterpretationThe protein subunit vaccine ZF2001 is well-tolerated and immunogenic. The safety and immunogenicity data from phase 1 and 2 trials for ZF2001 support the use of 25 g vaccine dose with three-dose schedule to an ongoing phase 3 large-scale evaluation for safety and efficacy. FundingNational Program on Key Research Project of China, National Science and Technology Major Projects of Drug Discovery, Strategic Priority Research Program of the Chinese Academy of Sciences, and Anhui Zhifei Longcom Biopharmaceutical.

Neutrophil-to-Lymphocyte Ratio Predicts Severe Illness Patients with 2019 Novel Coronavirus in the Early Stage

BackgroundSevere ill patients with 2019 novel coronavirus (2019-nCoV) infection progressed rapidly to acute respiratory failure. We aimed to select the most useful prognostic factor for severe illness incidence. MethodsThe study prospectively included 61 patients with 2019-nCoV infection treated at Beijing Ditan Hospital from January 13, 2020 to January 31, 2020. Prognostic factor of severe illness was selected by the LASSO COX regression analyses, to predict the severe illness probability of 2019-CoV pneumonia. The predictive accuracy was evaluated by concordance index, calibration curve, decision curve and clinical impact curve. ResultsThe neutrophil-to-lymphocyte ratio (NLR) was identified as the independent risk factor for severe illness in patients with 2019-nCoV infection. The NLR had a c-index of 0.807 (95% confidence interval, 0.676-0.38), the calibration curves fitted well, and the decision curve and clinical impact curve showed that the NLR had superior standardized net benefit. In addition, the incidence of severe illness was 9.1% in age [≥] 50 and NLR < 3.13 patients, and half of patients with age [≥] 50 and NLR [≥] 3.13 would develop severe illness. Based on the risk stratification of NLR with age, the study developed a 2019-nCoV pneumonia management process. ConclusionsThe NLR was the early identification of risk factors for 2019-nCoV severe illness. Patients with age [≥] 50 and NLR [≥] 3.13 facilitated severe illness, and they should rapidly access to intensive care unit if necessary.

COVID-19 cases from the first local outbreak of SARS-CoV-2 B.1.1.7 variant in China presented more serious clinical features: a prospective, comparative cohort study

BackgroundThe SARS-CoV-2 B.1.1.7 variant which was first identified in the United Kingdom (U.K.) has increased sharply in numbers worldwide and was reported to be more contagious. On January 17, 2021, a COVID-19 clustered outbreak caused by B.1.1.7 variant occurred in a community in Daxing District, Beijing, China. Three weeks prior, another non-variant (lineage B.1.470) COVID-19 outbreak occurred in Shunyi District, Beijing. This study aimed to investigate the clinical features of B.1.1.7 variant infection. MethodsA prospective cohort study was conducted on COVID-19 cases admitted to Ditan hospital since January 2020. Data of 74 COVID-19 cases from two independent COVID-19 outbreaks in Beijing were extracted as study subjects from a Cloud Database established in Ditan hospital, which included 41 Shunyi cases (Shunyi B.1.470 group) and 33 Daxing cases (Daxing B.1.1.7 group) that have been hospitalized since December 25, 2020 and January 17, 2021, respectively. We conducted a comparison of the clinical characteristics, RT-qPCR results and genomic features between the two groups. FindingsCases from Daxing B.1.1.7 group (15 [45.5%] male; median age, 39 years [range, 30.5, 62.5]) and cases from Shunyi B.1.470 group (25 [61.0%] male; median age, 31 years [range, 27.5, 41.0]) had a statistically significant difference in median age (P =0.014). Seven clinical indicators of Daxing B.1.1.7 group were significantly higher than Shunyi B.1.470 group including patients having fever over 38{degrees}C (14/33 [46.43%] in Daxing B.1.1.7 group vs. 9/41 (21.95%) in Shunyi B.1.470 group [P = 0 .015]), C-reactive protein ([CRP, mg/L], 4.30 [2.45, 12.1] vs. 1.80, [0.85, 4.95], [P = 0.005]), Serum amyloid A ([SAA, mg/L], 21.50 [12.50, 50.70] vs. 12.00 [5.20, 26.95], [P = 0.003]), Creatine Kinase ([CK, U/L]), 110.50 [53.15,152.40] vs. 70.40 [54.35,103.05], [P = 0.040]), D-dimer ([DD, mg/L], 0.31 [0.20, 0.48] vs. 0.24 [0.17,0.31], [P = 0.038]), CD4+ T lymphocyte ([CD4+ T, mg/L], [P = 0.003]), and Ground-glass opacity (GGO) in lung (15/33 [45.45%] vs. 5/41 [12.20%], [P =0.001]). After adjusting for the age factor, B.1.1.7 variant infection was the risk factor for CRP (P = 0.045, Odds ratio [OR] 2.791, CI [1.025, 0.8610]), SAA (0.011, 5.031, [1.459, 17.354]), CK (0.034, 4.34, [0.05, 0.91]), CD4+ T (0.029, 3.31, [1.13, 9.71]), and GGO (0.005, 5.418, [1.656, 17.729]) of patients. The median Ct value of RT-qPCR tests of the N-gene target in the Daxing B.1.1.7 group was significantly lower than the Shunyi B.1.470 group (P=0.036). The phylogenetic analysis showed that only 2 amino acid mutations in spike protein were detected in B.1.470 strains while B.1.1.7 strains had 3 deletions and 7 mutations. InterpretationClinical features including a more serious inflammatory response, pneumonia and a possible higher viral load were detected in the cases infected with B.1.1.7 SARS-CoV-2 variant. It could therefore be inferred that the B.1.1.7 variant may have increased pathogenicity. FundingThe study was funded by the National Key Research and Development Program (grant nos.2020YFC0846200 and 2020YFC0848300) and National Natural Science Foundation of China (grant no. 82072295).