Identification of pyruvate dehydrogenase kinase 1 inhibitors with anti-osteosarcoma activity.

Overexpression of pyruvate dehydrogenase kinases (PDKs), especially PDK1 has been observed in a variety of cancers. Thus, targeting PDK1 offers an attractive opportunity for the development of cancer therapies. In this letter, we reported the identification of two novel PDK1 inhibitors as anti-osteosarcoma agents. We found that TM-1 and TM-2 inhibited PDK1 with the IC50 values of 2.97 and 3.41 µM, respectively. Furthermore, TM-1 and TM-2 dose-dependently reduced phosphorylation of pyruvate dehydrogenase complex in MG-63 osteosarcoma cells. Finally, TM-1 and TM-2 were found to inhibit the proliferation of MG-63 cells with the EC50 values of 14.5, and 11.0 µM, respectively, meaning TM-1 and TM-2 could be promising leads for the discovery of potent PDK1 inhibitors.

Therapeutic Whole-body Hypothermia Protects Remote Lung, Liver, and Kidney Injuries after Blast Limb Trauma in Rats.

BACKGROUND: Severe blast limb trauma (BLT) induces distant multiple-organ injuries. In the current study, the authors determined whether whole-body hypothermia (WH) and its optimal duration (if any) afford protection to the local limb damage and distant lung, liver, and kidney injuries after BLT in rats. METHODS: Rats with BLT, created by using chartaceous electricity detonators, were randomly treated with WH for 30 min, 60 min, 3 h, and 6 h (n = 12/group). Rectal temperature and arterial blood pressure were monitored throughout. Blood and lung, liver, and kidney tissue samples were harvested for measuring tumor necrosis factor-α, interleukin-6 and interleukin-10, myeloperoxidase activity, hydrogen sulfide, and biomarkers of oxidative stress at 6 h after BLT. The pathologic lung injury and the water content of the lungs, liver, and kidneys and blast limb tissue were assessed. RESULTS: Unlike WH for 30 min, WH for 60 min reduced lung water content, lung myeloperoxidase activity, and kidney myeloperoxidase activity by 10, 39, and 28% (all P < 0.05), respectively. WH for 3 h attenuated distant vital organs and local traumatic limb damage and reduced myeloperoxidase activity, hydrogen peroxide and malondialdehyde concentration, and tumor necrosis factor-α and interleukin-6 levels by up to 49% (all P < 0.01). Likewise, WH for 6 h also provided protection to such injured organs but increased blood loss from traumatic limb. CONCLUSIONS: Results of this study indicated that WH may provide protection for distant organs and local traumatic limb after blast trauma, which warrants further study.

An Uncommon Case of Double-Hit Mantle Cell Lymphoma That Demonstrates a Transformation Process.

OBJECTIVES: Mantle cell lymphoma (MCL) is a mature B-cell lymphoma characterized by CCND1/IGH rearrangement. We reported a case of MCL harboring both CCND1/IGH and MYC/IGH rearrangements that also presented with an aggressive clinical course. METHODS: Biopsy specimens were evaluated by morphological staining, immunohistochemistry, flow cytometry, conventional cytogenetics, fluorescence in situ hybridization (FISH), and next-generation sequencing (NGS). RESULTS: Morphological and immunohistochemical staining of gallbladder samples demonstrated blastoid variant MCL. However, in the bone marrow sample, FISH indicated rearrangements in CCND1/IGH and MYC/IGH. Flow cytometry identified two groups of malignant lymphocytes. We sorted these two groups of cells. NGS then revealed that both cell types carried CCND1/IGH rearrangements and TP53 mutations. Furthermore, the CD19+/CD10+ cells carried additional MYC/IGH rearrangement and NOTCH2 mutation. CONCLUSIONS: The rearrangement of MYC and a mutation in NOTCH2 probably induced the transformation of MCL cells in this patient. This uncommon double-hit MCL case clearly demonstrates a transformation process.

The effect of qigong for pulmonary function and quality of life in patients with covid-19: A protocol for systematic review and meta-analysis.

BACKGROUND: Qigong is a traditional Chinese exercise method for health care, keeping fit and getting rid of diseases. It has the advantages of simple operation and few side effects. Corona Virus Disease 2019 (COVID-19) is an acute respiratory infectious disease caused by severe acute respiratory syndrome coronavirus 2(SARS-COV-2). Its clinical manifestations mainly include fever, fatigue, and dry cough. Clinical practice showed that Qigong had some therapeutic effects on pulmonary dysfunction caused by novel Coronavirus, but there was lacking in evidence of evidence-based medicine. The purpose of this protocol is to systematically evaluate the effects of Qigong on lung function and quality of life in COVID-19 patients, and to add evidence to evidence-based medicine for the clinical application of Qigong therapy. METHODS: Use computer to retrieve English database (PubMed, Embase, Web of Science, the Cochrane Library) and Chinese database (China Knowledge Network (CNKI), Wanfang Database, VIP Information Chinese Journal Service Platform (VIP), Chinese Biomedical Database). In addition, we manually retrieve randomized controlled clinical research from Baidu academic and Google academic from its establishment to July 2020. Two researchers independently extracted and evaluated the quality of the data included in the study, using RevMan5.3 to do meta-analyses of articles included, without language restrictions. RESULTS: This research evaluated the effectiveness and safety of Qigongs influence on patients pulmonary function and life quality by index such as 6-minute walk distance (6MWD), Forced expiratory volume in 1 second (FEV1), Forced vital capacity (FVC), Forced expiratory volume in 1 second/Forced vital capacity (FEV1/FVC), Forced expiratory volume in 1 second/prediction (FEV1/PRE), Self-rating anxiety scale (SAS), etc. CONCLUSIONS:: This study will provide reliable evidence-based evidence for the clinical application of Qigong in the treatment of COVID-19. PROSPERO REGISTRATION NUMBER: CRD42020191877.

Optimized multiparametric flow cytometric analysis of circulating endothelial cells and their subpopulations in peripheral blood of patients with solid tumors: a technical analysis.

BACKGROUND: Circulating endothelial cells (CECs) and their subpopulations could be potential novel biomarkers for various malignancies. However, reliable enumerable methods are warranted to further improve their clinical utility. This study aimed to optimize a flow cytometric method (FCM) assay for CECs and subpopulations in peripheral blood for patients with solid cancers. PATIENTS AND METHODS: An FCM assay was used to detect and identify CECs. A panel of 60 blood samples, including 44 metastatic cancer patients and 16 healthy controls, were used in this study. Some key issues of CEC enumeration, including sample material and anticoagulant selection, optimal titration of antibodies, lysis/wash procedures of blood sample preparation, conditions of sample storage, sufficient cell events to enhance the signal, fluorescence-minus-one controls instead of isotype controls to reduce background noise, optimal selection of cell surface markers, and evaluating the reproducibility of our method, were integrated and investigated. Wilcoxon and Mann-Whitney U tests were used to determine statistically significant differences. RESULTS: In this validation study, we refined a five-color FCM method to detect CECs and their subpopulations in peripheral blood of patients with solid tumors. Several key technical issues regarding preanalytical elements, FCM data acquisition, and analysis were addressed. Furthermore, we clinically validated the utility of our method. The baseline levels of mature CECs, endothelial progenitor cells, and activated CECs were higher in cancer patients than healthy subjects (P<0.01). However, there was no significant difference in resting CEC levels between healthy subjects and cancer patients (P=0.193). CONCLUSION: We integrated and comprehensively addressed significant technical issues found in previously published assays and validated the reproducibility and sensitivity of our proposed method. Future work is required to explore the potential of our optimized method in clinical oncologic applications.

Circulating endothelial cells and their subsets: novel biomarkers for cancer.

Angiogenesis contributes to the growth of solid tumors. Antiangiogenic agents are widely used in various cancers and considerable efforts have been made in the development of novel biomarkers that can predict the outcome of an anticancer treatment. Of those, circulating endothelial cells (CECs) and their subsets constitute a surrogate tool for monitoring disease activity. However, owing to the lack of standardization on the phenotypes and detection of CECs and their subsets, results have always been inconsistent and uninterpretable. In this review, we focus on the biological characteristics in terms of physiology, phenotypes and detection of CECs along with their subsets; review the current scenario of CEC enumeration as a surrogate biomarker in clinical oncology; and explore their future potential applications.

Discovery of human lactate dehydrogenase A (LDHA) inhibitors as anticancer agents to inhibit the proliferation of MG-63 osteosarcoma cells.

Human lactate dehydrogenase A (LDHA) has been identified as a potential therapeutic target in the area of cancer metabolism. Herein, we report the discovery of novel LDHA inhibitors through docking-based virtual screening and biological assays. The primary enzymatic assay suggested that compound 11 targeted LDHA with an IC50 value of 0.33 µM. The in vitro cytotoxic assay demonstrated that compound 11 reduced the growth of MG-63 cancer cells with an EC50 value of 3.35 µM. Finally, we found that compound 11 induced the apoptosis of MG-63 cancer cells in a dose dependent manner, upregulated the oxygen consumption rate (OCR), and decreased the lactate formation and extracellular acidification rate (ECAR) in MG-63 cancer cells. Collectively, our data suggested that compound 11 could be a promising lead for the development of potent LDHA inhibitors.

Transforming growth factor ß plays an important role in enhancing wound healing by topical application of Povidone-iodine.

Povidone-iodine (PVI) is principally used as an antimicrobial agent. It has been found that 0.5% PVI can attenuate congestion, edema and pain induced by pressure sores. Thus this study aimed to assess the effects of 0.5% PVI on acute skin wounds. Four full-thickness excisional wounds were generated on the dorsal skin of male Sprague-Dawley rats with a 10-mm sterile punch. Two wounds were left untreated and the other two were dressed with gauze with 0.5% PVI for 1 hour per day for the first 5 days after injury. 10-mm full-thickness excisional wounds were also generated on the dorsal skin of rats treated with 10 mg/kg SB431542 and all wounds were treated with 0.5% PVI for 5 days. PVI treatment enhanced wound healing via promotion of expression of α SMA and TGF ß, neovascularization and re-epithelialization. Interleukin 6 was reduced following PVI treatment. Inhibition of TGF ß abolished the effect of PVI treatment on wound closure. These data show that topical application of 0.5% PVI could promote acute skin wound healing though increased expression of TGF ß leading to enhanced formation of granulation tissue, even in the absence of obvious infection.

[The effect of murine mesenchymal stem cells on the differentiation and function of allogenetic bone marrow-derived dendritic cells.].

AIM: To study the effect of murine mesenchymal stem cells (MSCs) on the differentiation, maturation and function of allogenetic bone marrow-derived dendritic cells (DCs) and to investigate the mechanism of MSCs displaying immunoregulatory activity. METHODS: BALB/c mice BMCs were isolated and cultured in vitro and then coclutured with C57BL/6 murine bone marrows cells (BMCs) at different ratios in vitro to induce DCs generation. The phenotype of cells was analyzed by flow cytometry. Endocytosis was measured as the cellular uptake of fluorescein isothiocyanate (FITC)-dextram amd was quantified by flow cytometry. The level of IL-12 secretion in the cell culture supernatants was detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: Decreased expression of CD11c, CD14, CD83, CD86 and I-A(b);, down-regulated endocytosis capacity and interleukin-12 (IL-12) secretion of DCs were all observed when MSCs cocultured with BMCs at a higher ratio (1:10). CONCLUSION: Murine MSCs could supress the differentiation and maturation of DCs derived from allogeneic BMCs in vitro.