Background and objectives: Hepatic stellate cell (HSC) activation is the cardinal factor due to the accumulation of extracellular matrix proteins during the development of liver fibrosis. The aim of the present study was to find new targets for developing drugs to treat liver fibrosis, by screening the key genes involved in the activation of hepatic stellate cells. Methods: Differentially expressed genes were identified through TCGA database. RT-PCR, immunohistochemistry (IHC) assay, western blot, and ELISA were performed to evaluate the expression levels of FAT10 and fibrotic molecules. In vitro experiments were conducted to investigate the signaling pathways and biological functions of FAT10 in LX-2 cell lines. Results: In the present study, expression profiles obtained from the Gene Expression Omnibus (GEO) were used to explore the different genes expression between HSCs treated with or without carbon tetrachloride (CCl4). Human leukocyte antigen (HLA)-F adjacent transcript 10 (FAT10) was selected for further investigations. In animal model of carbon tetrachloride-induced liver fibrosis, the expression of FAT10 on activated HSCs is upregulated. In vitro, silencing FAT10 reduced TGF-β1-induced ECM activation and accumulation in LX-2 cells, and also suppressed the inflammatory response of LX-2 cells. Further Transwell results suggested that knockdown of FAT10 could inhibit TGF-β1-induced LX-2 cell migration and invasion. Mechanistically, FAT10 promotes its fibrotic activity through regulating sirtuin 1 (SIRT1), with a concomitant activation of ECM. Conclusions: These findings indicated an unexpected role of FAT10 in liver fibrosis development, suggesting that silencing FAT10 might represent a new strategy for the treatment of fibrotic liver diseases.

Most cardiomyocytes (CMs) in the adult mammalian heart are either binucleated or contain a single polyploid nucleus. Recent studies have shown that polyploidy in CMs plays an important role as an adaptive response to physiological demands and environmental stress and correlates with poor cardiac regenerative ability after injury. However, knowledge about the functional properties of polyploid CMs is limited. In this study, we generated tetraploid pluripotent stem cells (PSCs) by fusion of murine embryonic stem cells (ESCs) and somatic cells isolated from bone marrow or spleen and performed a comparative analysis of the electrophysiological properties of tetraploid fusion-derived PSCs and diploid ESC-derived CMs. Fusion-derived PSCs exhibited characteristics of genuine ESCs and contained a near-tetraploid genome. Ploidy features and marker expression were also retained during the differentiation of fusion-derived cells. Fusion-derived PSCs gave rise to CMs, which were similar to their diploid ESC counterparts in terms of their expression of typical cardiospecific markers, sarcomeric organization, action potential parameters, response to pharmacologic stimulation with various drugs, and expression of functional ion channels. These results suggest that the state of ploidy does not significantly affect the structural and electrophysiological properties of murine PSC-derived CMs. These results extend our knowledge of the functional properties of polyploid CMs and contribute to a better understanding of their biological role in the adult heart.

The aim of the current study was to investigate the anti-lung cancer effects of astragalin. Studies were also undertaken to evaluate its effects on apoptosis induction, ROS production, cellular migration and invasion and JAK/STAT3 signalling pathway. MTT assay was used to evaluate cell viability in NSCLC A549 cells after exposure to astragalin molecule. Apoptosis was investigated using AO/EB staining, comet assay and western blotting assay. Fluorescence microscopy was implemented to estimate ROS production. Cell migration and invasion were measured using transwell chambers assay. Effects of astragalin on JAK/STAT pathway were investigated using western blotting assay. Results showed astragalin molecule induced inhibition of proliferation in A549 cells in a dose-dependent fashion. Further, the antiproliferative effects were found to mediate via apoptosis as suggested by AO/EB staining and western blotting assay. Astragalin modulated the expressions of caspase-3, caspase-9, Bax, Bak, Cyt-c Bcl-2, XIAP and Bcl-xL. Astragalin induced DNA damage in A549 cells which too indicated apoptotic cell death. Astragalin molecule enhanced the production of ROS by A549 cells. It inhibited both cell migration and invasion of A549 cells in a concentration -dependent manner. Finally, astragalin drug was observed with remarkable potential of targeting JAK/STAT pathway in A549 NSCLC cells. These results indicated that astragalin drug could prove helpful in lung cancer treatment and research provided more in-vivo studies are performed.

Aberrant microRNA (miRNA) expression plays a critical role in osteosarcoma (OS) pathogenesis.In this study,we elucidated the involvement of miR-487a in OS and the underlying molecular mechanisms.We found that miR-487a was upregulated in OS clinical samples and cell lines.Knockdown of miR-487a suppressed OS cell growth and invasion and induced apoptosis;however,overexpression of miR-487a promoted OS cell growth and invasion.Accordingly,downregulation of miR-487a significantly suppressed tumor growth of OS xenografts in vivo.Furthermore,B-cell translocation gene 2 (BTG2) mRNA was found to be a novel target of miR-487a.Knockdown of BTG2 using small interfering RNA (siRNA) recapitulated the oncogenic effects of miR-487a,whereas BTG2 overexpression partially reversed these effects.Finally,miR-487a levels were found to be negatively correlated with BTG2 expression in OS clinical samples.Collectively,our data suggest that miR-487a is an oncogenic miRNA in OS and it lowers BTG2 expression.

Background: Coronavirus disease 2019 (COVID-19), a newly erupted respiratory infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has swept across the most of countries. The laboratory characteristics of COVID-patients accompanied with cancer and the risk factors for disease progression and survival of this particular population were few reported. Methods: We enrolled 585 confirmed COVID-19 patients admitted to our hospitals with measured interleukin-6 level on admission. Laboratory tests and outcome were extracted from electronic medical records. Data was divided to cancer group and non-cancer group to explorer the risk factors of progression and survival.Findings: A total of 44 patients with different cancer type (cancer group) and 541 patients without cancer (non-cancer group) were included. Cancer group had significant higher levels of NEUT, NLR, IL-6, and CRP than non-cancer group, but lymphocyte count and ALB were lower. Cancer group showed significantly higher progression rate (42·1% vs 22·5%) and mortality (27·27% vs 11·91%) than non-cancer group. Elevated IL-6 and CRP were the risk factors associated with progression among moderate patients and death in-hospital (all p<0·05) in non- cancer group. This correlation was not observed in caner group.Interpretation: IL-6, CRP, NEUT, and NLR were elevated in COVID-19 patients with cancer, with lower level of LYMP and ALB. IL-6 and CRP were positively correlated with progression and poor outcome in patients without cancer. As one of combined diseases, despite malignancy history did not directly affect the prognosis of COVID-19, but it could play a role in the poorer outcome through release of IL-6 and CRP.

Doxorubicin is a commonly used anthracycline chemotherapeutic agent; however, its application is limited owing to its cardiotoxicity. Current clinical treatments cannot efficiently or fully prevent doxorubicin-induced toxicity, primarily because its pathogenesis and mechanisms of action remain unknown. In this study, we established a rat model of chronic doxorubicin-induced cardiotoxicity, in which the severity of cardiac fibrosis and hydroxyproline levels increased in a time-dependent manner. Doxorubicin damaged the mitochondria and blood vessels and induced autophagy. Cells undergoing endothelial-to-mesenchymal transition (EndoMT)and those expressing endothelial cell and myofibroblast markers were simultaneously observed in vitro and in rats treated with doxorubicin. The NF-κB pathway was activated during EndoMT, andp65 and p-p65 were strongly expressed in the nucleus of endothelial cells in vitro. Taken together, these results suggest that vascular injury and cardiac fibrosis are characteristic symptoms of doxorubicin-induced cardiotoxicity. The NF-κB pathway-associated EndoMT may influence the pathogenesis of doxorubicin-induced cardiotoxicity, and the constituents of this pathway may be potential therapeutic targets to prevent the development of this condition.

Background: This study aimed to explore the correlation of integrin-α7 (ITGA7) with common cancer stem cell marker (CD44 and CD133) expressions, clinicopathological characteristics and survival profiles in non-small cell lung cancer (NSCLC) patients. Methods: Two hundred and seventy NSCLC patients who underwent resection were reviewed in this study and the expressions of ITGA7, CD44 and CD133 were detected using immunohistochemistry (IHC) assay in tumor tissue specimens. The clinical data (including age, gender, pathological grade, tumor size, lymph node metastasis (LYN), and TNM stage), survival data [including disease-free survival (DFS) and overall survival (OS)] were collected. Results: The numbers of NSCLC patients with ITGA7 high expression, CD44 high expression and CD133 high expression were 170 (63.0%), 241 (89.3%) and 37 (13.7%) respectively, and ITGA7 expression was positively associated with CD44 expression and CD133 expression. ITGA7 high expression was associated with higher pathological grade, larger tumor size, LYN and elevated TNM stage. However, there was no correlation of ITGA7 expression with age or gender. As for survival, patients with ITGA7 high expression presented reduced DFS and OS compared with those with ITGA7 low expression. In addition, multivariate Cox’s proportional hazards regression model analyses exhibited that ITGA7 high expression, higher pathological grade and LYN were independent risk factors for DFS and OS in NSCLC patients. Conclusions: ITGA7 expression positively correlates with CD44 and CD133 expressions, and its high expression associates with advanced tumor features as well as poor survivals in NSCLC.

Background Kimura’s disease (KD) is a rare chronic inflammatory disease with unknown etiology. It usually manifests as a painless soft tissue mass or subcutaneous nodule on one side of the patient’s head and/or neck and rarely affects multiple parts of the body. The disease is more common among young Asian males. Case presentation A 57-year-old Chinese woman complained of multiple masses on her body surface. Ultrasonography was used to examine the retroperitoneal, bilateral neck, bilateral supraclavicular, bilateral axillary, and bilateral inguinal superficial lymph nodes. Enlargement of multiple lymph nodes was found in all areas. Many solid nodules were also found in the right parotid gland and right posterior neck area, respectively. Numerous solid nodules were seen on the left chest wall. Laboratory tests showed that the percentage of eosinophils in the whole blood was 39.40%, total immunoglobulin E (IgE) level was > 5000 kU/L, and serum special IgE to Phadiatop (inhaled allergens) and fx5 (food allergens) were 1.01 and 1.04 kUA/L, respectively. After a complete examination, the masses located in the right neck, retroauricular and left axillary regions, and left chest wall were resected directly. Postoperative pathological findings revealed KD. Conclusions The case discussed in this study is extremely rare and did not meet the common affected areas and age characteristics of KD. This presentation can be used to improve disease awareness among physicians.

We report here a transgenic murine induced pluripotent stem cell (iPSC) line expressing puromycin N-acetyltransferase (PAC) and enhanced green fluorescent protein (EGFP) under the control of α-myosin heavy chain promoter. This transgenic cell line reproducibly differentiates into EGFP-expressing cardiomyocytes (CMs) which can be generated at high purity with puromycin treatment and exhibit molecular and functional properties of immature heart muscle cells. This genetically modified iPSC line can be used for assessment of the utility of CMs for myocardial repair, pharmacological and toxicological applications and development of improved cardiac differentiation protocols.

Ischemic heart disease is the main cause of death in western countries and its burden is increasing worldwide. It typically involves irreversible degeneration and loss of myocardial tissue leading to poor prognosis and fatal outcome. Autologous cells with the potential to regenerate damaged heart tissue would be an ideal source for cell therapeutic approaches. Here, we compared different methods of conditional culture for increasing the yield and cardiogenic potential of murine skeletal muscle-derived stem cells. A subpopulation of nonadherent cells was isolated from skeletal muscle by preplating and applying cell culture conditions differing in support of cluster formation. In contrast to static culture conditions, dynamic culture with or without previous hanging drop preculture led to significantly increased cluster diameters and the expression of cardiac specific markers on the protein and mRNA level. Whole-cell patch-clamp studies revealed similarities to pacemaker action potentials and responsiveness to cardiac specific pharmacological stimuli. This data indicates that skeletal muscle-derived stem cells are capable of adopting enhanced cardiac muscle cell-like properties by applying specific culture conditions. Choosing this route for the establishment of a sustainable, autologous source of cells for cardiac therapies holds the potential of being clinically more acceptable than transgenic manipulation of cells.

Long QT syndromes (LQTS) are heritable diseases characterized by prolongation of the QT interval on an electrocardiogram, which often leads to syncope and sudden cardiac death. Here we report the generation of induced pluripotent stems (iPS) cells from two patients with LQTS type 3 carrying a different point mutation in a sodium channel Nav1.5 (p.V240M and p.R535Q) and functional characterization of cardiomyocytes (CM) derived from them. The iPS cells exhibited all characteristic properties of pluripotent stem cells, maintained the disease-specific mutation and readily differentiated to CM. The duration of action potentials at 50% and 90% repolarization was longer in LQTS-3 CM as compared to control CM but this difference did not reach statistical significance due to high variations among cells. Sodium current recordings demonstrated longer time to peak and longer time to 90% of inactivation of the Na+ channel in the LQTS-3 CM. This hints at a defective Na+ channel caused by deficiency in open-state inactivation of the Na+ channel that is characteristic of LQTS-3. These analyses suggest that the effect of channel mutation in the diseased CM is demonstrated in vitro and that the iPS cell-derived CM can serve as a model system for studying the pathophysiology of LQTS-3, toxicity testing and design of novel therapeutics. However, further improvements in the model are still required to reduce cell-to-cell and cell line-to-cell line variability.

Background/Aims: Induced pluripotent stem (iPS) cells generated from accessible adult cells of patients with genetic diseases open unprecedented opportunities for exploring the pathophysiology of human diseases in vitro. Catecholaminergic polymorphic ventricular tachycardia type 1 (CPVT1) is an inherited cardiac disorder that is caused by mutations in the cardiac ryanodine receptor type 2 gene (RYR2) and is characterized by stress-induced ventricular arrhythmia that can lead to sudden cardiac death in young individuals. The aim of this study was to generate iPS cells from a patient with CPVT1 and determine whether iPS cell-derived cardiomyocytes carrying patient specific RYR2 mutation recapitulate the disease phenotype in vitro. Methods: iPS cells were derived from dermal fibroblasts of healthy donors and a patient with CPVT1 carrying the novel heterozygous autosomal dominant mutation p.F2483I in the RYR2. Functional properties of iPS cell derived-cardiomyocytes were analyzed by using whole-cell current and voltage clamp and calcium imaging techniques. Results: Patch-clamp recordings revealed arrhythmias and delayed afterdepolarizations (DADs) after catecholaminergic stimulation of CPVT1-iPS cell-derived cardiomyocytes. Calcium imaging studies showed that, compared to healthy cardiomyocytes, CPVT1-cardiomyocytes exhibit higher amplitudes and longer durations of spontaneous Ca2+ release events at basal state. In addition, in CPVT1-cardiomyocytes the Ca2+-induced Ca2+-release events continued after repolarization and were abolished by increasing the cytosolic cAMP levels with forskolin. Conclusion: This study demonstrates the suitability of iPS cells in modeling RYR2-related cardiac disorders in vitro and opens new opportunities for investigating the disease mechanism in vitro, developing new drugs, predicting their toxicity, and optimizing current treatment strategies.

Several types of terminally differentiated somatic cells can be reprogrammed into a pluripotent state by ectopic expression of Klf4, Oct3/4, Sox2, and c-Myc. Such induced pluripotent stem (iPS) cells have great potential to serve as an autologous source of cells for tissue repair. In the process of developing iPS-cell-based therapies, the major goal is to determine whether differentiated cells derived from iPS cells, such as cardiomyocytes (CMs), have the same functional properties as their physiological in vivo counterparts. Therefore, we differentiated murine iPS cells to CMs in vitro and characterized them by RT-PCR, immunocytochemistry, and electrophysiology. As key markers of cardiac lineages, transcripts for Nkx2.5, alphaMHC, Mlc2v, and cTnT could be identified. Immunocytochemical stainings revealed the presence of organized sarcomeric actinin but the absence of mature atrial natriuretic factor. We examined characteristics and developmental changes of action potentials, as well as functional hormonal regulation and sensitivity to channel blockers. In addition, we determined expression patterns and functionality of cardiac-specific voltage-gated Na+, Ca2+, and K+ channels at early and late differentiation stages and compared them with CMs derived from murine embryonic stem cells (ESCs) as well as with fetal CMs. We conclude that iPS cells give rise to functional CMs in vitro, with established hormonal regulation pathways and functionally expressed cardiac ion channels; CMs generated from iPS cells have a ventricular phenotype; and cardiac development of iPS cells is delayed compared with maturation of native fetal CMs and of ESC-derived CMs. This difference may reflect the incomplete reprogramming of iPS cells and should be critically considered in further studies to clarify the suitability of the iPS model for regenerative medicine of heart disorders.