Outbreaks of the Chikungunya virus (CHIKV) infection has been documented in over 40 countries, resulting in clinical symptoms characterized by fever and joint pain. Diagnosing CHIKV in a clinical lab setting is often omitted because of the high lab safety requirement. An infection system that mimics CHIKV infection will permit clinical evaluation of the production of neutralizing antibody for both disease diagnostics and treatment. We generated a CHIKV construct expressing CHIKV structural proteins. This construct permits the production of CHIKV pseudo-viral particles with a luciferase reporter. The pseudo-virus was able to infect a wide range of cell lines. The pseudovirus could be neutralized by the addition of neutralizing antibodies from patients. Taken together, we have developed a powerful system that can be handled at biosafety level 2 laboratories for evaluation of existence of CHIKV neutralizing antibodies.

Background: To investigate the epidemiology of Klebsiella pneumoniae (K. pneumoniae) inducing pyogenic liver abscess (PLA) in east China and the role of hypervirulent carbapenem-resistant K. pneumoniae (Hv-CRKP). Methods: Forty-three K. pneumoniae strains were collected from 43 patients with PLA at Hangzhou, China in 2017. Antimicrobial susceptibility tests, string test, multilocus sequence typing, pulsed-field gel electrophoresis, mobile genetic elements typing, regular PCR and sequencing, and Galleria mellonella (G. mellonella) lethality test were used to elucidate the epidemiology. Clinical data were collected. Results: K. pneumoniae strains with serotypes K1 and K2 accounted for 69.8%, which shared 46.5% and 23.3% respectively. K. pneumoniae strains with clonal group 23 were predominant with a rate of 34.9%. Such antimicrobials showed susceptible rates over 80.0%: cefuroxime, cefotaxime, gentamycin, ticarcillin/clavulanate, ceftazidime, cefoperazone/tazobactam, cefepime, aztreonam, imipenem, meropenem, amikacin, tobramycin, ciprofloxacin, levofloxacin, doxycycline, minocycline, tigecycline, chloramphenicol, and trimethoprim-sulfamethoxazole. PFGE dendrogram showed 29 clusters for the 43 K. pneumoniae strains. Three Hv-CRKP strains were confirmed by G. mellonella lethality test, showing a constituent ratio of 7.0% (3/43). Totally three deaths were found, presenting a rate of 7.0% (3/43). The three died patients were all infected with Hv-CRKP. Conclusions: K1 and K2 are the leading serotypes of K. pneumoniae causing PLA, which show highly divergent genetic backgrounds. Aminoglycosides, Generation 2^nd to 4^th cephalosporins, β-lactamase/β-lactamase inhibitors, carbapenems, fluoroquinolones are empirical choices. Hv-CRKP may confer an urgent challenge in the future.

Streptococcus pneumoniae is one major cause of pneumonia in human and contains various virulence factors that contribute to pathogenesis of pneumococcal disease. This study investigated the role of pneumolysin, Ply, in facilitating S. pneumoniae invasion into the host blood stream. S. pneumoniae strains were isolated from clinical blood and sputum samples and confirmed by PCR. Expression of ply gene was assessed by infecting human monocytes and pneumocytes. A total of 23 strains of S. pneumoniae isolated from blood (n = 11) and sputum (n = 12) along with S. pneumoniae ATCC49619 were used to infect human monocyte (THP-1) and Type II pneumocyte (A549) cell lines. All clinical strains of S. pneumoniae showed higher expression of ply mRNA than the American Type Culture Collection (ATCC) strain. Among the clinical strains, blood isolates showed higher expression of ply genes than sputum isolates, i.e., 2^1.5- to 2^1.6-folds in THP-1 and 2^0.4- to 2^4.9-folds in A549 cell lines. The data from the current study demonstrated that ply gene of blood- and sputum-derived S. pneumoniae is differentially expressed in two different cell lines. Under survival pressure, ply is highly expressed in these two cell lines for blood-derived S. pneumoniae, indicating that ply gene may facilitate S. pneumoniae invasion into the host blood system.

Background: To investigate the secretion of interleukin‐1β (IL‐1β), IL‐6, IL‐10, IL‐8, and soluble intercellular adhesin molecule 1 (sICAM‐1) from THP‐1 monocytes stimulated by different Streptococcus pneumoniae (S pneumoniae) strains.Methods: Fifty‐eight strains of S pneumoniae were collected: ATCC49619, 23 from sputum (sd‐SP), 23 from blood (bd‐SP), and 11 from cerebrospinal fluid (CSF; cd‐SP). Such strains were cultured and suspended at 0.5 McFarland. THP‐1 monocytes were cultured and resuspended at 5.0 × 10⁸/L, which were stimulated by S pneumoniae for 4, 8, and 12 hours, respectively. The suspensions were analyzed for IL‐1β, IL‐6, IL‐10, IL‐8, and sICAM‐1 using an ELISA method. The data were assayed with SPSS 19.0.Results: Contrary to IL‐10, the concentrations of IL‐1β, IL‐6, IL‐8, and sICAM‐1 all increased first and then decreased. IL‐1β and sICAM‐1 levels in the ATCC49619 group were both higher than all the clinical S pneumoniae groups (sd‐SP, bd‐SP, and cd‐SP), IL‐6 and IL‐8 versa, and IL‐10 equal. The difference among clinical S pneumoniae groups lay only in sICAM‐1. cd‐SP group showed lower sICAM‐1 concentrations than sd‐SP and bd‐SP groups at both 4 and 8 hours. However, they became equal at 12 hours.Conclusions: The secretion summit is 8 hours for IL‐1β, IL‐6, IL‐8, and sICAM‐1, bottom for IL‐10. Different clinical S pneumoniae strains show similar ability to induce THP‐1 cells secreting interleukins. However, cd‐SP induces THP‐1 cells secreting lower sICAM‐1 than sd‐SP and bd‐SP, which may in turn facilitate its invasion into CSF.

BackgroundCOVID-19 has been circulating worldwide since December 2019. However, its independent risk factors of mortality need further insights in elderly critical COVID-19 patients.MethodsTotally 48 elderly and critically ill COVID-19 patients with clear end point were enrolled when the data were collected, with 16 discharged and 32 died. Kaplan-Meier analysis and Cox regression were performed to identify the risk factors of mortality in elderly critical COVID-19 patients. Survival curve was conducted to present the impact of Diabetes Mellitus on mortality. Mann-Whitney U and t tests were used to 39 clinical variates between the survivor and non-survivor groups.ResultsAs Kaplan-Meier analysis confirmed, only three variates Diabetes Mellitus, chronic obstructive pulmonary disease and family aggregation showed significant difference between the survivor and non-survivor groups. However, only variate Diabetes Mellitus presented significance in Cox regression. Higher C-reaction protein, interleukin-2 (IL-2), IL-8 and creatinine were detected in survivor group than non-survivor group, which was reverse to estimated glomerular filtration rate. The other laboratory finding showed no significant difference between survivor and non-survivor groups.ConclusionsDiabetes Mellitus, chronic obstructive pulmonary disease and family aggregation can contribute to mortality by COVID-19 while Diabetes Mellitus also reduces the survival time of elderly and critically ill COVID-19 patients, highlighting the more important role of Diabetes Mellitus. Laboratory findings could not serve as good predictors of death for elderly and critically ill COVID-19 patients.