Clinical and Translational Medicine

INTRODUCTIONElevated D-dimer is a predictor of severity and mortality in COVID-19 patients and heparin use during in-hospital stay has been associated with decreased mortality. COVID-19 patient autopsies have revealed thrombi in the microvasculature, suggesting intravascular coagulation as a prominent feature of organ failure in these patients. Interestingly, in COVID-19, pulmonary compliance is preserved despite severe hypoxemia corroborating the hypothesis that perfusion mismatch may play a significant role in the development of respiratory failure. METHODSWe describe a series of 27 consecutive COVID-19 patients admitted to Sirio-Libanes Hospital in Sao Paulo-Brazil and treated with heparin in therapeutic doses tailored to clinical severity. RESULTS AND DISCUSSIONPaO2/FiO2 ratio increased significantly over the 72 hours following the start of anticoagulation, from 254({+/-}90) to 325({+/-}80), p=0.013, and 81% of the patients were discharged home within a mean time of 11.4 ({+/-}7.9) days. Most mechanically ventilated patients (67%) were extubated within 12.5({+/-} 5.7) days. There were no bleeding complications or fatal events. Even though this uncontrolled case series does not offer absolute proof of DIC as the underlying mechanism of respiratory failure in COVID-19, patients positive response to tailored dose heparinization contributes to the understanding of the pathophysiological mechanism of the disease and provides valuable information for the treatment of these very sick patients while we await the results of further prospective controlled studies.

Critically ill COVID-19 patients have relatively well-preserved lung mechanics despite severe gas exchange abnormalities, a feature not consistent with classical ARDS but more consistent with pulmonary vascular disease. Patients with severe COVID-19 also demonstrate markedly abnormal coagulation, with elevated D-dimers and higher rates of venous thromboembolism. We present four cases of patients with severe COVID-19 pneumonia with severe respiratory failure and shock who demonstrated immediate improvements in gas exchange and/or hemodynamics with systemic tPA. Subject category4.6 ICU Management and Outcome

We detected active alveolar regrowth in the lung of a 58-year-old COVID-19 patient who underwent lung transplantation due to severe lung hemorrhage. Specifically, immunohistological and scanning electronic microscopy analyses revealed that alveolar type II epithelial cells (AT2 cells) accumulate in response to viral pneumonia and that these AT2 cells actively proliferate and differentiate into squamous AT1-like alveolar epithelial cells. Thus, our work establishes that alveolar regrowth does occur in post-COVID-19 injury adult human lungs.

Gain of chromosome 1q locus is a common cytogenetic feature associated with intracranial ependymomas; however, candidate non-coding RNAs on this locus have not been identified. Recent studies have reported somatic copy number alterations for long non coding RNA (lncRNA) FAL1/FALEC residing on chromosome 1q to stabilize BMI1, an epigenetic repressor and PRC1 component, leading with to downregulation of p21/CDKN1A tumor suppressor gene. We aimed to study the role of FAL1 in ependymomas, its association with 1q gain, BMI1/p21 regulatory axis and clinicopathological parameters. Using SNP array analysis (GSE32101), 31% (discovery cohort) and 63.8% (in-house cohort) showed amplification/gain of FAL1 locus with higher prevalence in intracranial tumors. Copy number gain of FAL1 locus was significantly associated with increased FAL1 (p=0.003) and BMI1 (p=0.007) levels, and reduced p21 (p=0.001) expressions. Interestingly, gain of FAL1 locus and FAL1 transcripts did not show any association with 1q gain or RELA fusions. Subcellular localization reported FAL1 to be expressed in nuclear compartment in ependymomas. Chromatin immunoprecipitation-qPCR demonstrated in-vivo binding of BMI1 at p21 promoter locus with BMI1 target genes to be enriched in cell architecture related pathways. A 3-tier survival analysis between FAL1 gain and increased expression levels of FAL1 and BMI1 correlated with poor outcome in our cohort. Ours is the first study demonstrating gain of FAL1 locus and its interplay with the BMI1/p21 axis in intracranial ependymomas. Further studies into this epigenetic regulatory mechanism will unravel novel driver mutations in intracranial ependymomas HighlightsSomatic variations in enhancer long non-coding RNA has been recently attributed for various clinical malignancies including cancers. Gain of 1q locus is a common cytogenetic variation observed in intracranial ependymomas. Our study has demonstrated, focal amplification of enhancer lncRNA mapping to Chromosome 1q, FAL1/FALEC, to be involved in oncogenicity/ progression of ependymomas. Moreover, our data suggests a positive association with BMI1 (a PRC1 component) with FAL1 levels, indicating downregulation of BMI1 target gene involved in cell cycle, p21. Furthermore, a 3-tier prognosis analysis (between FAL1 gain, FAL1 and BMI1 expressions) suggests a negative survival outcome. Our study highlights the importance of somatic variation in non-coding genome with ependymoma survival.

BackgroundPatients with sepsis-induced acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) commonly suffer from severe pulmonary thrombosis, but clinical trials of anti-coagulant therapies in sepsis and ARDS patients have failed. ARDS patients with thrombocytopenia also exhibit increased mortality, and widespread pulmonary thrombosis is often seen in coronavirus disease 2019 (COVID-19) ARDS patients. MethodsEmploying different amounts of microbeads to induce various levels of pulmonary thrombosis. Acute lung injury was induced by either lipopolysaccharide i.p. or cecal ligation and puncture. Endothelial cell (EC)-targeted nanoparticle coupled with CDH5 promoter was employed to delivery plasmid DNA expressing the CRISPR/Cas9 system for EC-specific gene knockout or expressing Alox15 for EC-specific overexpression. Additionally, thrombocytopenia was induced by genetic depletion of platelets using DTRPf4Cre mice by breeding Pf4Cre mice into the genetic background of DTR mice. ResultsWe show that while severe pulmonary thrombosis or thrombocytopenia augments sepsis-induced ALI, the induction of mild pulmonary thrombosis conversely reduces endothelial cell (EC) apoptosis, ALI, and mortality via sustained expression of endothelial arachidonate 15-lipoxygenase (Alox15). Endothelial Alox15 knockout via EC-targeted nanoparticle delivery of CRISPR/Cas9 plasmid DNA in adult mice abolished the protective impact of mild lung thrombosis. Conversely, overexpression of endothelial Alox15 inhibited the increases in ALI caused by severe pulmonary thrombosis. The clinical relevance of the findings was validated by the observation of reduced ALOX15-expressing ECs in lung autopsy samples of ARDS patients. Additionally, restoration of pulmonary thrombosis in thrombocytopenic mice also normalized endotoxemia-induced ALI. ConclusionWe have demonstrated that moderate levels of thrombosis protect against sepsis-induced inflammatory lung injury via endothelial Alox15. Overexpression of Alox5 inhibits severe pulmonary thrombosis-induced increase of ALI. Thus, activation of ALOX15 signaling represents a promising therapeutic strategy for treatment of ARDS, especially in sub-populations of patients with thrombocytopenia and/or severe pulmonary thrombosis.

BackgroundSchizandrin A is major components extracted from Schisandra chinenzis-Turcz. Baill and Schisandra sphenanthear Rend. etWils. Schizandrin A exhibits remarkable hepatoprotective, antiviral and anti-inflammatory effects. However, the anti-tumor effects and its molecular mechanism were still known limited. MethodsThe affinity between Schizandrin A and COX-2/ALOX5 protein was analyzed using network pharmacology, computer molecular docking, and surface plasmon resonance experiments. Bioinformatic analysis and review of clinical characteristics were conducted to assess the necessity of simultaneous blocking of COX-2 and ALOX5 in pancreatic cancer (PC). LC/MS metabolomics and RNA-sequencing were utilized to investigate the effects of schizandrin A on the activation and expression of COX-2/ALOX5 in PC cells. Biological function experiments were conducted to investigate the inhibitory effects of Schizandrin A on PC cell proliferation and cancer-associated fibroblast activation in vitro and in vivo. ResultsSchizandrin A demonstrated a high affinity for binding directly with COX-2 and ALOX5, with kinetic association constants of 14.8 M and 21.8 M, respectively. PC exhibited a significant COX-2/ALOX5 signature, while PC cases with a high COX-2/ALOX5 signature showed lower overall survival and disease-free survival rates. Treatment of PC cells with schizandrin A resulted in decreased COX-2/ALOX5 activity and expression, leading to inhibition of leukotriene and prostaglandin production, as well as suppression of the downstream pathway NF-kappaB signaling. Schizandrin A demonstrated significant inhibitory effects on the proliferation and sphericity of PC cells in vitro, as well as on cell proliferation in vivo, while exhibiting low toxicity to normal tissues. Treatment of conditioned medium from PC cells with schizandrin A resulted in reduced induction of normal fibroblasts into cancer-associated fibroblasts. Furthermore, mutations in the binding sites of ALXO5 (Arg246) and COX-2 proteins (Ile124 and Ser126) resulted in a significant decrease in affinity to Schizandrin A, and blocking the inhibitory effects of schizandrin A. ConclusionsTaken together, schizandrin A directly bound with COX-2 and ALOX5, reduced their activation and leukotrienes and prostaglandins production, thus exhibiting distinguished effects on suppressing PC proliferation and inhibiting the ability of PC cell to induce normal fibroblasts to transform into tumor-associated fibroblasts. Therefore, schizandrin A represents a potentially novel therapeutic approach for PC.

Dyslipidemia is a hallmark of obstructive sleep apnea (OSA)-induced metabolic syndrome, yet the mechanisms remain poorly understood. We conducted a genome-wide association study on lipid traits in the OSA cohorts, identifying the SNP rs3745683 in ANGPTL8, significantly associated with reductions in multiple lipid traits. ANGPTL8, an essential lipogenic hormone and potential therapeutic target for metabolic syndrome, showed elevated expression in OSA patients compared to healthy controls, strongly correlated with increased insulin levels. Notably, ANGPTL8 expression can be upregulated by insulin stimulation, indicating it as an insulin-responsive hormone regulating dyslipidemia in OSA. Mechanistically, SNP rs3745683 attenuated ANGPTL8 transcription by inhibiting its binding to transcription factor YBX1. Insulin prompted AKT1 to phosphorylate YBX1 at Ser102, facilitating YBX1s nuclear translocation and subsequent regulation of ANGPTL8 expression and lipid synthesis. Specific knockdown of YBX1 in mouse liver confirmed its necessity for ANGPTL8 expression and hepatic lipid synthesis in vivo. Our findings highlight ANGPTL8 as a critical regulator of dyslipidemia in OSA patients, offering a promising therapeutic avenue for managing metabolic syndrome in OSA.

Head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy with high mortality rates, often exhibiting resistance to conventional treatments such as radiotherapy (RT) or a combination of chemotherapy and radiotherapy (CRT). The nerve growth factor receptor (NGFR, also known as p75NTR or CD271) is a well-established cancer stem cell marker in melanoma, where it has been linked to resistance to multiple therapies. In HNSCC, NGFR has been reported as a poor prognostic marker, with its overexpression associated with disease progression. However, its contribution to therapy resistance in HNSCC remains unknown. Here, we show in a cohort of RT/CRT-treated patients that NGFR expression identifies individuals with poor prognosis and increased risk of recurrence following standard RT/CRT. Moreover, we found that NGFR is upregulated in the human Detroit 562 cisplatin (CDDP)-resistant HNSCC cell line in vitro and in vivo. Functional studies demonstrated that genetic knock out of NGFR in these cisplatin-resistant cells restored sensitivity to CDDP in vivo. These results indicate that NGFR contributes to cisplatin resistance in HNSCC. NGFR is upregulated in tumors from patients with poorer prognosis and an increased risk of recurrence after standard radiotherapy and/or RT/CRT, as well as in cisplatin-resistant models. Altogether, our findings open the way to consider NGFR as a new potential therapeutic target to overcome or mitigate cisplatin resistance in HNSCC.

PurposeAHNAK2 has been reported as tumor promoting protein by mediating tumor cell invasion and metastasis in a variety of malignancies, but the role of AHNAK2 in GC is indistinct. MethodsImmunohistochemistry(IHC)was used to verify in GC tissues, clinical and pathological files were collected to figure out the correlation of AHNAK2 and prognosis in GC patients. GC cell lines were cultured to detect the expression and location of AHNAK2 by westernblot and immunofluorescence. Knockdown AHNAK2 to observe the invasion and metastasis of GC cell. Transcriptome sequencing was performed to explore the potential regulatory mechanism of AHNAK2. ResultsIHC results shows that AHNAK2 is upregulated in GC patients compared to normal people, overall survival in highly expressed AHNAK2 is poor,and AHNAK2 expression was positively correlated with lymph node metastasis. The expression of AHNAK2 in tumor cells is higher than in normal cell. Knockdown AHNAK2 decreased the proliferation, invasion, migration ability of GC cells, and increased cell apoptosis. Transcriptome sequencing reveals that AHNAK2 mediates GC progression by regulating Wnt/{beta}-catenin axis. ConclusionOur findings suggest that AHNAK2 may promote the progression of GC by regulating Wnt/{beta}-catenin signaling pathway.

As a novel genetic biomarker, the potential role of SH3D21 in hepatocellular carcinoma remains unclear. Here, we decipher the expression and function of SH3D21 in human hepatocellular carcinoma. The expression level and clinical significance of SH3D21 in hepatocellular carcinoma patients, the relationship between SH3D21 and the features of tumor microenvironment (TME) and role of SH3D21 in promoting hepatocellular carcinoma progression were analyzed based on the bulk samples obtained from The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) databases. Single-cell sequencing samples from Gene Expression Omnibus (GEO) database were employed to verify the prediction mechanism. Additionally, different biological effects of SH3D21 on hepatocellular carcinoma cells were investigated by qRT-PCR, CCK-8 assay, colony forming assay and Western blot analysis. Bioinformatics analysis and in vitro experiments revealed that the expression level of SH3D21 was up-regulated in hepatocellular carcinoma and correlated with the poor prognosis in hepatocellular carcinoma patients. SH3D21 effectively promoted the proliferation, invasion, and migration as well as the formation of immunosuppressive microenvironment of hepatocellular carcinoma. In addition, SH3D21 can activate the PI3K/AKT/mTOR signaling pathway. SH3D21 stimulates the progression of hepatocellular carcinoma by activating the PI3K/AKT/mTOR signaling pathway, and SH3D21 can serve as a prognostic biomarker and therapeutic target for hepatocellular carcinoma.

According to studies, numerous chemotherapeutic drugs can facilitate programmed cell death via pyroptosis. Clarifying the mechanism by which cisplatin kills gastric cancer cells is crucial for enhancing gastric cancers sensitivity to chemotherapy and elucidating the mechanism of drug resistance in gastric cancer. The differentially expressed genes following cisplatin treatment were identified using second-generation sequencing technology. Bioinformatics was used to investigate the functional enrichment of differentially expressed genes and core genes in tumor cells killed by cisplatin. Cox regression analyses were used to examine the pyroptosis core genes that worked as independent prognostic factors for patients with gastric cancer. The expression of core genes in gastric cancer cells was silenced by siRNA, and the changes in the proliferation of gastric cancer cells were observed. The expression of related genes and the survival of gastric cancer cells after the addition of cisplatin were observed. The second-generation sequencing, RT-PCR and Western blotting showed that the pyroptosis core gene was significantly highly expressed after cisplatin treatment. The results of differential gene enrichment of cisplatin-treated gastric cancer cells showed that differential genes were mainly concentrated in biological processes and signaling pathways related to pyroptosis. GSDME protein is highly expressed after cisplatin treatment, and it is also a poor prognostic factor for gastric cancer patients and an independent prognostic factor. After the same dose of cisplatin treatment, the survival rate of siGSDME gastric cancer cells was significantly higher than that of GSDME regular expression gastric cancer cells. After acting on gastric cancer cells, cisplatin triggers pyroptosis by stimulating the activation of genes such as GSDME, resulting in the death of gastric cancer cells. GSDME is an independent prognostic factor for gastric cancer patients and is significantly linked with a shorter OS. In gastric cancer cells, silencing GSDME can substantially reduce cisplatins cytotoxicity.

Systemic inflammation in critically ill patients can lead to serious consequences such as acute respiratory distress syndrome (ARDS), a condition characterized by the presence of lung inflammation, edema, and impaired gas exchange, associated with poor survival. Understanding molecular pathobiology is essential to improve critical care of these patients. To this end, we use multimodal profiles of SARS-CoV-2 infected hospitalized participants to the Biobanque Quebecoise de la COVID-19 (BQC-19) to characterize endophenotypes associated with different degrees of disease severity. Proteomic, metabolomic, and genomic characterization supported a role for neutrophil-associated procoagulant activity in severe COVID-19 ARDS that is inversely correlated with sphinghosine-1 phosphate plasma levels. Fibroblast Growth Factor Receptor (FGFR) and SH2-containing transforming protein 4 (SHC4) signaling were identified as molecular features associated with endophenotype 6 (EP6). Mechanical ventilation in EP6 was associated with alterations in lipoprotein metabolism. These findings help define the molecular mechanisms related to specific severe outcomes, that can be used to identify early unfavorable clinical trajectories and treatable traits to improve the survival of critically ill patients.

The human gene microcephalin (MCPH1) plays a key role in DNA damage-induced cellular responses and chromosome condensation. Recent clinical studies proposed MCPH1 as a tumor suppressor gene in lung cancer, pancreatic cancer, and breast cancer, yet its roles remain poorly understood in other types of tumors. Pan-cancer analyses of MCPH1 are urgently required to help us understand the potential molecular functions of MCPH1 in other types of tumors. Here, we used several bioinformatic database and tools, including TCGA, GEO, ONCOMINE, and Human Protein Atlas to investigate the role of MCPH1 in 33 tumor types. We found that the expression of MCPH1 in tumor cases and normal cases were significantly different, and the higher expression of MCPH1 generally predicted poor overall survival for tumor patients, such as acute myeloid leukemia, liver hepatocellular carcinoma, and pancreatic adenocarcinoma. Meanwhile, lower expression of the MCPH1 gene was related to poor OS prognosis for KIRC and gastric cancer. Moreover, the expression level of MCPH1 was highly associated with the immune microenvironment. Our result provides some fresh light into the oncogenic roles of MCPH1 in various human cancers and revealed that MCPH1 may be a potential diagnostic and prognostic marker in LAML, PAAD, and gastric cancer.

ObjectiveS-Adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are indicators of global transmethylation and may play an important role as markers of severity of COVID-19. MethodsThe levels of plasma SAM and SAH were determined in patients admitted with COVID-19 (n = 56, mean age = 61). Lung injury was identified by computed tomography (CT) in accordance with the CT0-4 classification. ResultsSAM was found to be a potential marker of lung damage risk in COVID-19 patients (SAM > 80 nM; CT3,4 vs. CT 0-2: relative ratio (RR) was 3.0; p = 0.0029). SAM/SAH > 6.0 was also found to be a marker of lung injury (CT2-4 vs. CT0,1: RR = 3.47, p = 0.0004). Interleukin-6 (IL-6) levels were associated with SAM ({rho} = 0.44, p = 0.01) and SAH ({rho} = 0.534, p = 0.001) levels. ConclusionsHigh SAM levels and high methylation index are associated with the risk of lung injury in COVID-19 patients. The association of SAM and SAH with IL-6 indicates an important role of transmethylation in the development of cytokine imbalance in COVID-19 cases.

In contrast to early reports of conventional acute respiratory distress syndrome (ARDS) as the underlying pathophysiology of hypoxemic respiratory failure observed in patients with severe COVID-19, more recent findings implicate direct involvement of the pulmonary vasculature in giving rise to these symptoms. In earlier research, we demonstrated that patients with COVID-19 showed markedly reduced pulmonary blood volumes in pulmonary vessels <5 mm2 in cross-sectional area visible on imaging (termed "BV5"), with attendant dilation of larger, more proximal vessels. Here, we present preliminary results in which reduced BV5 is shown to correlate significantly with increased need for supplemental oxygen and abnormal arterial blood gas measurements in hospitalized COVID-19 patients. We suggest a potential mechanistic link between observed clinical, pathological, and imaging findings, and outline how these may be helpful in clinical assessment as well as the development of novel therapies.

ObjectivesMost mesenchymal tumors found in the uterine corpus are benign tumors; however, uterine leiomyosarcoma is a malignant tumor with unknown risk factors that repeatedly recurs and metastasizes. In some cases, the histopathologic findings of uterine leiomyoma and uterine leiomyosarcoma are similar and surgical pathological diagnosis using excised tissue samples is difficult. It is necessary to analyze the risk factors for human uterine leiomyosarcoma and establish diagnostic biomarkers and treatments. Female mice deficient in the proteasome subunit low molecular mass peptide 2 (LMP2)/{beta}1i develop uterine leiomyosarcoma spontaneously. MethodologyOut of 334 patients with suspected uterine mesenchymal tumors, patients diagnosed with smooth muscle tumors of the uterus were selected from the pathological file. To investigate the expression status of biomarker candidate factors, immunohistochemical staining was performed with antibodies of biomarker candidate factors on thin-cut slides of human uterine leiomyosarcoma, uterine leiomyoma, and other uterine mesenchymal tumors. ResultsIn human uterine leiomyosarcoma, there was a loss of LMP2/{beta}1i expression and enhanced cyclin E1 and Ki-67 expression. In human uterine leiomyomas and normal uterine smooth muscle layers, enhanced LMP2/{beta}1i expression and the disappearance of the expression of E1 and Ki-67 were noted. The pattern of expression of each factor in other uterine mesenchymal tumors was different from that of uterine leiomyosarcoma. ConclusionsLMP2/{beta}1i, cyclin E1, and Ki-67 may be candidate factors for biomarkers of human uterine leiomyosarcoma. Further large-cohort clinical trials should be conducted to establish treatments and diagnostics for uterine mesenchymal tumors.

Alarmins are endogenous molecules that alert the immune system to circumvent the invading antigens in the host. They are basically sensed by specific cellular receptors, leading to recruitment and activation of dendritic cells to mount innate and adaptive immune responses. However, a dual role of alarmins as mediators in either antitumor immunity or tumor progression has been also reported. Here, we investigated the status of the HMGB-1 alarmin and the alarmin-like NPM/B23 in 162 debut NSCLC patients and 60 age-matched healthy individuals as control. NSCLC patients exhibited lower median values of circulating HMGB-1 compared to control (30.04 versus 37.30 ng/ml, p=0.02), but higher levels of HMGB-1 were positively correlated with tumor size > 2.0 cm (p=0.004) irrespectively of tumor histology. Otherwise, circulating NPM/B23 levels were significantly increased in NSCLC patients respect to control (812 versus 551 pg/ml, p=0.00027). Likewise, patients with tumors > 2 cm exhibited higher levels NPM/B23. Strong association was observed between HMGB-1 and NPM/B23 in serum of NSCLC patients (r=0.679 p=0.0001) compared to healthy individuals who exhibited a weaker correlation (r=0.278 p=0.031). However, the strongest correlation between both alarmins was observed in patients with STAS and tumors [&le;] 2 cm (r=0.900 p=0.0001). Co-expression of HMGB-1 and NPM/B23 was also observed in tumor tissues although to a lesser extent. Our data unveil that concomitant circulating HMGB-1 and NPM/B23 could serve as a putative alarmin-based biomarker of NSCLC disease progression.

Patients with tuberous sclerosis complex (TSC) develop an array of multi-organ disease manifestations, with angiomyolipomas (AML) and cysts in the kidneys being one of the most common and deadly. Early and regular AML/cyst detection and monitoring are vital in lowering TSC patient morbidity and mortality. However, current standard of care for imaging-based methods are not designed for rapid screening, posing challenges for early detection. To identify potential diagnostic screening biomarkers of AML/cysts, we performed global untargeted metabolomics in blood samples from 283 kidney AML/cyst-positive or -negative TSC patients using mass spectrometry. We identified seven highly sensitive chemical features, including octanoic acid, that predict kidney AML/cysts in TSC patients. Patients with elevated octanoic acid have lower levels of very long-chain fatty acids (VLCFAs), suggesting that dysregulated peroxisome activity leads to overproduction of octanoic acid via VLCFA oxidation. This study highlights serum metabolites as novel biomarkers for TSC kidney tumor diagnosis and offers valuable metabolic insights into the disease.

BackgroundPathogenesis of Coronavirus disease 2019 (Covid-19) is poorly understood. Most histologic studies come from post-mortem analysis, with existing data indicating that histologic features of acute respiratory distress syndrome are typically present in fatal cases. However, this observation may be misleading, due to confounding factors in pre-terminal disease, including injury resulting from prolonged mechanical ventilation. Ante-mortem lung biopsy may provide major pathogenetic insights, potentially providing a basis for novel treatment approaches. AimThis comparative, multicenter, prospective, observational study was planned to identify ante-mortem histological profile and immunohistochemical features of lung tissue in patients with Covid-19 in early and late phases of the disease, including markers of inflammatory cells and major pathways involved in the cytokine storm triggering. MethodsEnrolled patients underwent lung biopsy, according to the study protocol approved by local Ethical Committee, either within 15 days of the first symptoms appearing (early phase) or after >15 days (more advanced disease). Key exclusion criteria were excessive or uncorrectable bleeding risk and cardiovascular disease with heart failure. Lung samples were obtained by conventional transbronchial biopsy, trans-bronchial lung cryobiopsy or surgical lung biopsy. Results23 patients were enrolled: 12 patients underwent lung biopsy within 15 days and 11 patients more than 15 days after the onset of symptoms. Early biopsies were characterized by spots of patchy acute lung injury (ALI) with alveolar type II cells hyperplasia and significant vascular abnormalities (disordered angiogenesis with alveolar capillary hyperplasia, luminal enlargement and thickened walls of pulmonary venules, perivascular CD4-T-cell infiltration), with no hyaline membranes. In the later stages, the alveolar architecture appeared disrupted, with areas of organizing ALI, venular congestion and capillary thromboembolic microangiopathy. Striking phenotypic features were demonstrated in hyperplastic pneumocytes and endothelial cells, including the expression of phospho-STAT3 and molecules involved in immunoinhibitory signals (PD-L1 and IDO-1). Alveolar macrophages exhibited macrophage-related markers (CD68, CD11c, CD14) together with unusual markers, such as DC-Lamp/CD208, CD206, CD123/IL3AR. ConclusionA morphologically distinct "Covid pattern" was identified in the earlier stages of the disease, with prominent epithelial and endothelial cell abnormalities, that may be potentially reversible, differing strikingly from findings in classical diffuse alveolar damage. These observations may have major therapeutic implications, justifying studies of early interventions aimed at mitigating inflammatory organ injury.

Non-small cell lung cancer (NSCLC), a subtype of lung cancer, affects millions of people. While chemotherapy and other treatments have improved, the 5 year survival rate of NSCLC patients is still only 21%. Early diagnosis is essential for increasing survival as treatments have higher effectiveness at earlier stages of NSCLC. Noninvasive blood-based liquid biopsy tests for NSCLC may be useful for diagnosis and prognosis. MicroRNA (miRNA) and messenger RNA present in blood can serve as biomarkers for such tests. The present study identified 13 miRNAs that are underexpressed in the tissue and blood of NSCLC patients using Gene Expression Omnibus data. Following Kaplan Meier analysis, miR-140-3p, miR-29c, and miR-199a were selected as candidate biomarkers and demonstrated statistically significant prognostic power. An ROC analysis of miR-140-3p expression between NSCLC patients and controls had an area under curve value of 0.85. Functional enrichment analysis of the miRNA target genes revealed several overrepresented pathways relevant to cancer. Eight target genes were hub genes of the protein protein interaction network and possessed significant prognostic power. A combination of IL6, SNAI1, and CDK6 achieved a hazard ratio of 1.4 with p < 0.001. These biomarkers are especially valuable because they can be identified in blood and reflect the tumor state. Since all miRNAs were underexpressed in both tissue and blood, detecting expression of a biomarker miRNA in blood provides information on its expression in tissue as well. These miRNAs may be useful biomarkers for NSCLC prognostic and diagnostic tests and should be further studied.