Single cell analysis reveals inhibition of angiogenesis attenuates the progression of heterotopic ossification in Mkx^(−/−)mice

Tendon heterotopic ossification(HO)is characterized by bone formation inside tendon tissue,which severely debilitates people in their daily life.Current therapies fail to promote functional tissue repair largely due to our limited understanding of HO pathogenesis.Here,we investigate the pathological mechanism and propose a potential treatment method for HO.Immunofluorescence assays showed that the Mohawk(MKX)expression level was decreased in human tendon HO tissue,coinciding with spontaneous HO and the upregulated expression of osteochondrogenic and angiogenic genes in the tendons of Mkx^(−/−)mice.Single-cell RNA sequencing analyses of wild-type and Mkx^(−/−)tendons identified three cell types and revealed the excessive activation of osteochondrogenic genes during the tenogenesis of Mkx^(−/−)tendon cells.Single-cell analysis revealed that the gene expression program of angiogenesis,which is strongly associated with bone formation,was activated in all cell types during HO.Moreover,inhibition of angiogenesis by the small-molecule inhibitor BIBF1120 attenuated bone formation and angiogenesis in the Achilles tendons of both Mkx mutant mice and a rat traumatic model of HO.These findings provide new insights into the cellular mechanisms of tendon HO and highlight the inhibition of angiogenesis with BIBF1120 as a potential treatment strategy for HO.

Simultaneous virus identification and characterization of severe unexplained pneumonia cases using a metagenomics sequencing technique

Many viruses can cause respiratory diseases in humans.Although great advances have been achieved in methods of diagnosis,it remains challenging to identify pathogens in unexplained pneumonia(UP) cases.In this study,we applied next-generation sequencing(NGS) technology and a metagenomic approach to detect and characterize respiratory viruses in UP cases from Guizhou Province,China.A total of 33 oropharyngeal swabs were obtained from hospitalized UP patients and subjected to NGS.An unbiased metagenomic analysis pipeline identified 13 virus species in 16 samples.Human rhinovirus C was the virus most frequently detected and was identified in seven samples.Human measles virus,adenovirus B 55 and coxsackievirus A10 were also identified.Metagenomic sequencing also provided virus genomic sequences,which enabled genotype characterization and phylogenetic analysis.For cases of multiple infection,metagenomic sequencing afforded information regarding the quantity of each virus in the sample,which could be used to evaluate each viruses' role in the disease.Our study highlights the potential of metagenomic sequencing for pathogen identification in UP cases.

Clinical factors associated with composition of lung microbiota and important taxa predicting clinical prognosis in patients with severe community-acquired pneumonia

Few studies have described the key features and prognostic roles of lung microbiota in patients with severe community-acquired pneumonia(SCAP).We prospectively enrolled consecutive SCAP patients admitted to ICU.Bronchoscopy was performed at bedside within 48 h of ICU admission,and 16S rRNA gene sequencing was applied to the collected bronchoalveolar lavage fluid.The primary outcome was clinical improvements defined as a decrease of 2 categories and above on a 7-category ordinal scale within 14 days following bronchoscopy.Sixty-seven patients were included.Multivariable permutational multivariate analysis of variance found that positive bacteria lab test results had the strongest independent association with lung microbiota(R2=0.033;P=0.018),followed by acute kidney injury(AKI;R2=0.032;P=0.011)and plasma MIP-1βlevel(R2=0.027;P=0.044).Random forest identified that the families Prevotellaceae,Moraxellaceae,and Staphylococcaceae were the biomarkers related to the positive bacteria lab test results.Multivariable Cox regression showed that the increase inα-diversity and the abundance of the families Prevotellaceae and Actinomycetaceae were associated with clinical improvements.The positive bacteria lab test results,AKI,and plasma MIP-1βlevel were associated with patients’lung microbiota composition on ICU admission.The families Prevotellaceae and Actinomycetaceae on admission predicted clinical improvements.

Sirolimus combined with oseltamivir and corticosteroid treatment for a puerpera with severe pneumonia caused by 2009 pandemic H1N1:A case report

Severe pneumonia in patients infected with the 2009 pandemic H1N1(pH1N1)virus was partially attributed to excessive immune response.Anti-virus treatment for these patients was insufficient.Here we reported the therapy effect of sirolimus,an immunosuppressor,combined with oseltamivir and corticosteroid for a puerpera with severe pneumonia caused by pH1N1 virus.This patient has infected with the pH1N1 virus in late pregnancy,and antiviral therapy was not implemented timely.She developed severe pneumonia and ARDS rapidly and need receive a cesarean section on the 39th week after pregnancy.After giving birth to a healthy baby,she received a combination of oseltamivir,sirolimus and corticosteroid,and improved in the following days.Moreover,the cytokines in serum and viral loads in BALF decreased significantly.She recovered without infectious symptoms and was discharged.Sirolimus combined with oseltamivir and corticosteroid is likely responsible for lowering the viral loads,reducing the patient's cytokine level,and further improving her clinical outcomes.It provides evidence that adjuvant treatment was beneficial to patients with severe pneumonia induced by the pH1N1 virus.

The substitution V379I in PA protein attenuates the pathogenicity of influenza A(H1N1)pdm09 viruses in mice

Dear Editor,Since August 2010,the world was in the post-pandemic period and the influenza A(H1N1)pdm09(pdmH1N1)virus co-circulated with H3N2 and B virus seasonally(WHO,2010).The continuous surveillance on the pdmH1N1 virus in the post-pandemic period was recommended by WHO.In order to detect unusual events that might indicate a shift in severity disease,or emergence of a new virus,we collected a total of 312 throat swabs of laboratory-confirmed pdmHlNl patients from Hunan and Jilin provinces in

Characterization of two SARS-CoV-2 subgenomic RNA dynamics in severe COVID-19 patients

Little is known about Subgenomic RNA(sgRNA)dynamics in patients with Coronavirus diseases 2019(COVID-19).We collected 147 throat swabs,74 gut swabs and 46 plasma samples from 117 COVID-19 patients recruited in the LOTUS China trial(ChiCTR2000029308)and compared E and orf7a sgRNA load in patients with different illness duration,outcome,and comorbidities.Both sgRNAs were detected in all the three types of samples,with longest duration of 25,13,and 17 days for E sgRNA,and 32,28,and 17 days for orf7a sgRNA in throat,gut,and plasma,respectively.A total of 95%(57/60)of patients had no E sgRNA detected after 10 days post treatment,though 86%of them were still E RNA positive.High correlation on titer was observed between sgRNA encoding E and orf7a gene.sgRNA showed similar variation in the standard care and Lopinavir-Ritonavir group.Patients with diabetes and heart diseases showed higher pharyngeal E sgRNA at the first day(P=0.016 and 0.013,respectively)but no difference at five days after treatment,compared with patients without such commodities.Patients with hypertension and cerebrovascular diseases showed no difference in the pharyngeal sgRNA levels at both one and five days after treatment,compared with patients without these two commodities.E sgRNA levels in the initial infection showed no correlation with the serum antibody against spike,nucleoprotein,and receptor binding domains at ten days later.sgRNA lasted a long period in COVID-19 patients and might have little effect on humoral response.

Battling COVID-19 Using Lessons Learned from 100 Years of Fighting Against Influenza

In 1918,a letter from a doctor at an USA Army camp read:“These men start with what appears to be an ordinary attack of LaGrippe or Influenza,and when brought to the Hosp.they very rapidly develop the most vicious type of pneumonia that has ever been seen…and a few hours later you can begin to see the Cyanosis extending from their ears and spreading all over the face until it is hard to distinguish the colored men from the white.It is only a matter of a few hours then until death comes…It is horrible.One can stand it to see one,two or twenty men die,but to see these poor devils dropping like flies…We have been averaging about 100 deaths per day…Pneumonia means in about all cases death…We have lost an outrageous number of Nurses and Drs.It takes special trains to carry away the dead.For several days there were no coffins and the bodies piled up something fierce…It beats any sight they ever had in France after a battle.An extra long barracks has been vacated for the use of the Morgue,and it would make any man sit up and take notice to walk down the long lines of dead soldiers all dressed and laid out in double rows…Good By old Pal,God be with you till we meet again.”