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Abstract Calcific aortic valve disease (CAVD) and osteoarthritis (OA) are common diseases in the ageing population and share similar pathogenesis, especially in inflammation. This study aims to discover potential diagnostic and therapeutic targets in patients with CAVD and OA. Three CAVD datasets and one OA dataset were obtained from the Gene Expression Omnibus database. We used bioinformatics methods to search for key genes and immune infiltration, and established a ceRNA network. Immunohistochemical staining was performed to verify the expression of candidate genes in human and mice aortic valve tissues. Two key genes obtained, leucine rich repeat containing 15 (LRRC15) and secreted phosphoprotein 1 (SPP1), were further screened using machine learning and verified in human and mice aortic valve tissues. Compared to normal tissues, the infiltration of immune cells in CAVD tissues was significantly higher, and the expressions of LRRC15 and SPP1 were positively correlated with immune cells infiltration. Moreover, the ceRNA network showed extensive regulatory interactions based on LRRC15 and SPP1. The authors’ findings identified LRRC15 and SPP1 as hub genes in immunological mechanisms during CAVD and OA initiation and progression, as well as potential targets for drug development.

Even though N6-methyladenosine (m6A) RNA modifications are increasingly being implicated in human disease, their mechanisms are not fully understood in smokers with coronary artery disease (CAD). Thirty m6A-related regulators’ expression (MRRE) in CAD individuals (smokers and non-smokers) were analyzed from GEO. Support Vector Machine, random forest, and nomogram models were constructed to assess its clinical value. Consensus clustering, principal component analysis, and ssGSEA were used to construct a full picture of m6A-related regulators in smokers with CAD. Oxygen-glucose deprivation (OGD) and qRT-PCR were used to validate hypoxia’s effect on MRRE. A comparison between smokers with CAD and controls revealed lower expression levels of RBM15B, YTHDC2, and ZC3H13. Based on three key MRREs, all models showed good clinical value, and smokers with CAD were divided into two distinct molecular subgroups. The correlations were found between key MRRE and the degree of immune infiltration. Three key MRREs in HUVECs and FMC84 mouse cardiomyocytes were reduced in the OGD group. Through hypoxia, smoking might reduce the expression levels of RBM15B, YTHDC2, and ZC3H13 in smokers with CAD. Our findings provide an important theoretical basis for the treatment of smokers with CAD.

There are limited cases of heart-lung transplantation (HLT) in children worldwide owing to lack of donors, demanding surgical teamwork, and arduous post-operative management. Post-transplant management difficulties stem from the possible development of several post-operative complications, with infection being a common complication. Intracranial fungal infections are difficult to diagnose and prone to treatment delays because of their relatively insidious onset and atypical clinical presentation. Here, we present a case of a cerebral infection developed 3 months after HLT in a 10-year-old child, showing no positive results on conventional imaging or cerebrospinal fluid (CSF) examination and culture. On metagenomic next-generation sequencing of the cerebrospinal fluid, the causative organism was finally determined as Aspergillus. After administering 1-year anti-Aspergillus treatment, no recurrence of intracranial fungal infection was noted during the 3-year follow-up. This case illustrates the multifaceted diagnostic techniques for cerebral aspergillosis after HLT and shows the significance of dynamic monitoring of symptoms, such as headache, and of metagenomic sequencing results, trends in intracranial pressure and (1-3)-β-D-glucan levels for guiding diagnosis and treatment.

CglE is a putative dihydrolipoamide dehydrogenase (DLDH) that shares significant protein sequence homology (50% identity and 70% similarity) to the IbeA invasin contributing to the pathogenesis of E. coli meningitis. However, the biochemical, biological and pathogenic functions of CglE are unknown. In order to characterize the role of CglE in the pathogenesis of meningitic infections, infectomic, bioinformatics and molecular approaches were used to analyze and predict its structure and function. First, our comparative infectomic studies showed that CglE and IbeA are present in the genetic island GimA as a pair of homologous proteins that are encoded by two different operons, cgl (GimA2) and ibe (GimA4), at different locations. A similar pair of proteins is also present in Silicibacter sp which belongs to the most abundant and ecologically relevant marine bacterial groups. Meningitic E. coli K1 and Silicibacter sp have to survive under harsh environments (cerebrospinal fluid and ocean) with poor nutrition, suggesting that this pair of proteins is important for energy metabolism in the both microbes. Secondly, bioinformatic analysis indicated that CglE is a putative DLDH, which is the E3 component of pyruvate dehydrogenase complex. A FAD-binding domain and homologous flavoprotein regions are present in CglE. DLDH has been identified as virulence factors contributing to the pathogenesis of hepatitis C virus and pneumococcal infections. The sequence of CglE is homology to that of IbeA, an invasion protein of meningitic E. coli, and the surface probability and hydrophilicity are very similar to each other. Thirdly, the expression, purification and biochemical analysis of CglE protein was further carried out to determine whether CglE shares similar functions as IbeA and whether it is a new class of DLDH. The cglE gene was amplified by PCR and subcloned into pET22b(+) then expressed in E. coli BL21(DE3) as a fusion protein with His6 tag at its C-terminus induced by IPTG. CglE fusion protein was purified. Like IbeA, CglE is able to bind to an IbeA-binding protein, vimentin. In further studies, we will examine whether CglE is a new class of DLDH and how it contributes to the pathogenesis of meningitic infections.