TMEM166 inhibits cell proliferation, migration and invasion in hepatocellular carcinoma via upregulating TP53.

Transmembrane protein 166 (TMEM166), an endoplasmic reticulum-associated protein, functions in many diseases via regulating autophagy and/or apoptosis. However, the role of TMEM166 in hepatocellular carcinoma (HCC) remains largely unknown. In this study, we detected the expression of TMEM166 in HCC by real-time fluorescent quantitative PCR (RT-qPCR), immunohistochemistry and western blot. To investigate its biological function and underlying mechanism in HCC, TMEM166 was overexpressed in HCC cell lines and assessed its effects on cell proliferation, migration, invasion, apoptosis and cell cycle by MTT assay, wound healing assay, Transwell assay, Annexin V-FITC/PI assay, JC-1 staining and flow cytometry assay, respectively. Results demonstrated that the expression of TMEM166 was significantly decreased in HCC and was associated with advanced TNM clinical stage and poor clinical outcome of HCC patients. TMEM166 overexpression inhibited HCC cells proliferation, migration and invasion. Furthermore, TMEM166 inhibited cell proliferation by inducing apoptosis and cell cycle arrest via upregulating anti-oncogene TP53 and TP53 knockdown significantly alleviated the anti-tumor effects of TMEM166 on HCC cells. This study provides the first comprehensive analysis the role of TMEM166 in HCC. TMEM166 displays a fine anti-tumor activity on HCC cells involving a mechanism of upregulating TP53. This study suggests TMEM166 is a potential target for the treatment of HCC.

An Autophagy-Related Gene Signature Associated With Clinical Prognosis and Immune Microenvironment in Gliomas.

Glioma is one of the leading causes of death from cancer, and autophagy-related genes (ARGs) play an important role in glioma occurrence, progression, and treatment. In this study, the gene expression profiles and clinical data of glioma patients were obtained from The Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA), respectively. ARGs were obtained from the Human Autophagy Database. We analyzed the expression of the ARGs in glioma and found that 73 ARGs were differentially expressed in tumor and normal tissues. Univariate Cox regression analysis was used to identify prognostic differentially expressed ARGs (PDEARGs). Least absolute shrinkage and selection operator (LASSO) and multivariate Cox regression analyses were performed on the PDEARGs to determine the risk genes; and BRIC5, NFE2L2, GABARAP, IKBKE, BID, MAPK3, FKBP1B, MAPK8IP1, PRKCQ, CX3CL1, NPC1, HSP90AB1, DAPK2, SUPT20H, and PTEN were selected to establish a prognostic risk score model for TCGA and CGGA cohorts. This model accurately stratified patients with different survival outcomes, and the autophagy-related signature was also appraised as being an independent prognostic factor. We also constructed a prognostic nomogram using risk score, age, gender, WHO grade, isocitrate dehydrogenase (IDH) mutation status, and 1p/19q co-deletion status; and the calibration plots showed excellent prognostic performance. Finally, Pearson correlation analysis suggested that the ARG signature also played an essential role in the tumor immune microenvironment. In summary, we constructed and verified a novel autophagy-related signature that was tightly associated with the tumor immune microenvironment and could serve as an independent prognostic biomarker in gliomas.

Transplantation of ACE2- Mesenchymal Stem Cells Improves the Outcome of Patients with COVID-19 Pneumonia.

A coronavirus (HCoV-19) has caused the novel coronavirus disease (COVID-19) outbreak in Wuhan, China. Preventing and reversing the cytokine storm may be the key to save the patients with severe COVID-19 pneumonia. Mesenchymal stem cells (MSCs) have been shown to possess a comprehensive powerful immunomodulatory function. This study aims to investigate whether MSC transplantation improves the outcome of 7 enrolled patients with COVID-19 pneumonia in Beijing YouAn Hospital, China, from Jan 23, 2020 to Feb 16, 2020. The clinical outcomes, as well as changes of inflammatory and immune function levels and adverse effects of 7 enrolled patients were assessed for 14 days after MSC injection. MSCs could cure or significantly improve the functional outcomes of seven patients without observed adverse effects. The pulmonary function and symptoms of these seven patients were significantly improved in 2 days after MSC transplantation. Among them, two common and one severe patient were recovered and discharged in 10 days after treatment. After treatment, the peripheral lymphocytes were increased, the C-reactive protein decreased, and the overactivated cytokine-secreting immune cells CXCR3+CD4+ T cells, CXCR3+CD8+ T cells, and CXCR3+ NK cells disappeared in 3-6 days. In addition, a group of CD14+CD11c+CD11bmid regulatory DC cell population dramatically increased. Meanwhile, the level of TNF-α was significantly decreased, while IL-10 increased in MSC treatment group compared to the placebo control group. Furthermore, the gene expression profile showed MSCs were ACE2- and TMPRSS2- which indicated MSCs are free from COVID-19 infection. Thus, the intravenous transplantation of MSCs was safe and effective for treatment in patients with COVID-19 pneumonia, especially for the patients in critically severe condition.

Exosomes from mesenchymal stem/stromal cells: a new therapeutic paradigm.

Mesenchymal stem/stromal cells (MSCs) have been demonstrated to hold great potential for the treatment of several diseases. Their therapeutic effects are largely mediated by paracrine factors including exosomes, which are nanometer-sized membrane-bound vesicles with functions as mediators of cell-cell communication. MSC-derived exosomes contain cytokines and growth factors, signaling lipids, mRNAs, and regulatory miRNAs. Increasing evidence suggests that MSC-derived exosomes might represent a novel cell-free therapy with compelling advantages over parent MSCs such as no risk of tumor formation and lower immunogenicity. This paper reviews the characteristics of MSC exosomes and their fate after in vivo administration, and highlights the therapeutic potential of MSC-derived exosomes in liver, kidney, cardiovascular and neurological disease. Particularly, we summarize the recent clinical trials performed to evaluate the safety and efficacy of MSC exosomes. Overall, this paper provides a general overview of MSC-exosomes as a new cell-free therapeutic paradigm.

Perfusion Zones of Extended Transverse Skin Paddles in Muscle-Sparing Latissimus Dorsi Myocutaneous Flaps for Breast Reconstruction.

BACKGROUND: The authors report their experience using extended transversely oriented skin paddles in muscle-sparing latissimus dorsi pedicled flaps for breast reconstruction as an alternative to thoracodorsal artery perforator flaps. METHODS: A retrospective review was conducted of patients who underwent muscle-sparing latissimus dorsi flap pedicled breast reconstruction from January of 2009 to July of 2014 with at least 3-month follow-up. Surgical outcomes and complications were analyzed. RESULTS: Fifty-three patients underwent a total of 81 muscle-sparing latissimus dorsi pedicled flaps for breast reconstruction. Extended transversely oriented skin paddles ranged from 7 to 9 cm vertically by 25 to 35 cm horizontally and were perfused by a strip of latissimus dorsi muscle that was approximately 25 percent of the total muscular volume. Twenty patients had indocyanine green angiography revealing three distinct zones of perfusion in the extended transversely oriented skin paddles. The area of earliest perfusion (designated zone 1) was directly over the muscle containing the perforators. The second best area of perfusion (zone 2) was lateral to the muscle (toward the axilla). The last and relatively least well-perfused area (zone 3) was medial to the muscle (toward the spine). Zone 3 still had adequate viability. There were no flap losses. Minor complications included wound infection [six of 81 (7.4 percent)], fat necrosis [three of 81 (3.7 percent)], and seroma [four of 81 (4.9 percent)]. CONCLUSIONS: Muscle-sparing latissimus dorsi pedicled flaps with extended transversely oriented skin paddles are reliable alternatives to thoracodorsal artery perforator flaps for breast reconstruction. Three zones of perfusion were delineated in the extended transversely oriented skin paddles on indocyanine green imaging, and all three zones were viable. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.

Outcome of Dupuytren Contractures After Collagenase Clostridium Histolyticum Injection: A Single-institution Experience.

BACKGROUND: Collagenase clostridium histolyticum (CCH) injection is an alternative to surgery for patients with Dupuytren disease (DD) of the metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints. The success of surgical and nonsurgical treatment modalities for DD is reported to vary widely between 25% and 80% (J Bone Joint Surg Am. 1985;67:1439-1443; Plast Reconstr Surg. 2007;120:44e-54e; J Bone Joint Surg Am. 2007;89:189-198; J Hand Surg Am. 2011:36:936-942; J Hand Surg Am. 1990;15:755-761; J Hand Surg Br. 1996;21:797-800; J Bone Joint Surg Br. 2000;82:90-94; Plast Reconstr Surg. 2005;115:802-810; Ann Plast Surg. 2006;57:13-17). This study presents the outcomes of patients with DD contractures treated with CCH injections at a single institution. METHODS: An institutional review board-approved retrospective study was conducted of patients with DD of the hand treated with CCH injections in a single institution from February 2010 to April 2015. All patients received the recommended dose of 0.58 mg of CCH and returned for joint manipulation the following day. Data for follow-up at 7 and 30 days postoperatively and up to 5 years for patients who returned seeking further therapy for recurrent symptoms were reviewed. RESULTS: One hundred thirteen patients with a total of 146 affected joints (72 MCP; 74 PIP) were treated with CCH injections (95 males; 18 females; age, 40-92 y). Successful CCH therapy occurred in 75% of injected joints (109/146 joints; 59 MCP; 50 PIP), as defined by less than 5 degrees of contracture after treatment. Twenty-three percent of treated joints had partial correction (34/146 joints; 13 MCP; 21 PIP), as defined by between 5 and 30 degrees of residual contracture after treatment. Three patients (2%) had a failure of treatment, as defined by unchanged or worsened contracture from pretreatment baseline measurements. Fifteen patients (13%) returned to the clinic seeking additional therapy for recurrent joint contracture symptoms in 17 joints over a span of 1.5 months to 4 years after initial successful or partially successful treatment (17/143, 12%; 5 MCP; 12 PIP). Recurrence was defined as patients who sought treatment for a return of symptoms or greater than 20 degrees contracture in the setting of a palpable cord after initial full or partial contracture correction. DISCUSSIONS: Our 5-year outcome of CCH injections for DD contractures revealed full correction in 75% and partial correction in 23% of treated joints, with failure of treatment seen in only 2% of patients. Thirteen percent of the patients returned for additional treatment because of symptoms resulting from contracture recurrence in 12% of initially corrected or partially corrected joints. These positive outcomes are comparable with current surgical treatment modalities (J Hand Surg Am. 1990;15:755-761; J Bone Joint Surg Am. 1962;44B:602-613; J Clin Epidemiol. 2000;53:291-296). The use of CCH injections is an important nonsurgical treatment alternative for DD contractures of the MCP and PIP joints.

Low level laser (LLL) attenuate LPS-induced inflammatory responses in mesenchymal stem cells via the suppression of NF-κB signaling pathway in vitro.

BACKGROUND: Considering promising results in animal models and patients, therapeutic use of MSCs for immune disease is likely to undergo continued evaluation. Low-lever laser (LLL) has been widely applied to retard the inflammatory reaction. LLL treatment can potentially be applied in anti-inflammatory therapy followed by stem cell therapy. AIM OF THE STUDY: The purpose of this study was to investigate the effect of LLL (660 nm) on the inflammatory reaction induced by LPS in human adipose derived mesenchymal stem cells (hADSCs) and pertinent mechanism. MATERIALS AND METHODS: Anti-inflammatory activity of LLL was investigated by LPS-induced mesenchymal stem cells. The production and expression of pro-inflammatory cytokines were evaluated by ELISA kits and RT-qPCR. Nuclear translocation of NF-κB was indicated by immunofluorescent staining. Phosphorylation status of NF-κB p65 and IκBα were illustrated by western blot assay. ROS generation was measured with CM-H2DCFDA, and NO secretion was determined by DAF-FM. We studied surface expression of lymphocyte activation markers when Purified peripheral blood mononuclear cell (PBMC) were activated by phytohaemagglutinin (PHA) in the presence of 3 types of treated MSCs. RESULTS: LLL reduced the secretion of IL-1ß, IL-6, IL8, ROS and NO in LPS treated MSCs. Immunofluorescent assay demonstrated the nuclear translocation decrease of NF-κB in LLL treated LPS induced MSCs. Western blot analysis also suggested that LLL suppressed NF-κB activation via regulating the phosphorylation of p65 and IκBα. MSC significantly reduced the expression of activation markers CD25 and CD69 on PHA-stimulated lymphocytes. CONCLUSION: The results indicate that LLL suppressed the activation of NF-κB signaling pathway in LPS treated MSCs through inhibiting phosphorylation of p65 and IκBα, which results in good anti-inflammatory effect. In addition, LLL attenuated activation-associated markers CD25 and CD69 in co-cultures of PBMC and 3 types of treated MSCs.

The Muscle-Sparing Latissimus Dorsi Flap for Breast Reconstruction: A Retrospective Review of 126 Consecutive Flaps.

Our experience in the use of muscle-sparing latissimus dorsi (MSLD) flaps for breast reconstruction is presented. The procedure was performed on 83 patients by the senior author over an 8-year period. Of the 83 patients reviewed, a total of 126 MSLD flaps were done for immediate (26) or delayed (100) breast reconstructions. Preoperative and postoperative photographs were taken of all patients, and complications as well as ancillary procedures were recorded. The MSLD flap is shown to be a versatile option for breast reconstruction in a variety of clinical settings, with minimal complications and satisfactory aesthetic results.

Low-Level Laser Effect on Proliferation, Migration, and Antiapoptosis of Mesenchymal Stem Cells.

Mesenchymal stem cells (MSCs) have been proved to be an important element in cell-based therapy. Photobiomodulation used extremely low-level lasers (LLLs) to affect the behavior of cells. The effect mechanism of LLLs on MSCs from human remained to be discovered. In this study, cell viability was assessed using MTS assays and cell cycle was evaluated by fluorescence-activated cell sorting (FACS). The influence of LLLs on mitochondrial biogenesis (fission or fusion) and function (ATP, reactive oxygen species [ROS], nitric oxide [NO]) was evaluated by transmission electron microscope, FACS, quantitative real time polymerase chain reaction (q-PCR), and immunocytochemistry. Cell migration and cytoskeleton alteration (actin and tubulin) were evaluated using transwell assay, immunocytochemistry, enzyme-linked immunosorbent assay, and western blotting. Cell apoptosis was evaluated using FACS, immunocytochemistry, and western blotting. We investigated that certain influence of LLLs on MSCs in vitro 6 or 24 h after 1 h of LLL irradiation. The mechanism of the effects included proliferation rate increase mediated by increased S phase proportion; mitochondrial biogenesis and function alteration mediated by fusion (Mfn1, Mfn2, and Opa-1) and fission (Fis1, Drp-1, and MTP18)-related proteins, NRF1, TFAM, PGC-1a, and upregulated intracellular ROS and NO concentration; migration acceleration through the ERK1/2 and FAK pathway and upregulation of growth factors such as HGF and PDGF; and resistance to apoptosis with increased Bcl-2 and decreased Bax, or through tunneling nanotube formation between LLL-treated MSCs and 5-fluorouracil-induced apoptotic MSCs. These observations suggested that LLLs enhanced stem cell survival and therapeutic function, which could appear to be an innovative pretreatment in the application of MSCs.

Mesenchymal Stem Cells and Cell Therapy for Bone Repair.

Mesenchymal stem cells (MSCs) represent a new therapeutic paradigm for a number of diseases because they possess unique biological characteristics such as multipotency, immunomodulation and production of cytokines. Currently, 425 MSC based clinical trials have been conducted for at least 12 kinds of pathological conditions, with many completed trials demonstrating the safety and efficacy of MSCs. Here, we provide an overview of the clinical status of MSCs by searching the public clinical trials database http://clinicaltrials.gov. Particularly, the role of MSCs in clinical trials to treat bone defects and injuries is highlighted.