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In patients with localized scleroderma (LoS), facial deformity induced by subcutaneous atrophy greatly reduces life quality. Autologous fat grafting (AFG) is used for volume restoration but with low‐fat retention due to various reasons. Adipose‐derived stem cells (ADSCs) have shown potential effects in improving fat retention. We aimed to compare the feasibility and efficacy of improving fat retention in LoS patients among the ADSCs‐assisted, the stromal vascular fraction (SVF)‐assisted and conventional AFG methods. A pilot study with a 6‐month follow‐up among 18 LoS patients was conducted. Participants were randomly assigned into three AFG groups: conventional group, SVF‐assisted group, and ADSCs‐assisted group. The SVF‐assisted group received SVF‐assisted AFG at the SVF:fat ratio of 1:1. The ADSCs‐assisted group received the mixture of ADSCs‐enriched fat graft supplemented with 5 × 105 ADSCs/mL fat. Volume retention was measured by magnetic resonance imaging, and clinical photographs were taken for outcome evaluation. At sixth‐month follow‐up, the fat retention of ADSCs‐assisted group was 49.83 ± 3.61%, significantly higher than 31.75 ± 1.73% of SVF‐assisted group (P = .0004), and 21.86 ± 1.68% of the conventional group (P, Adipose‐derived stem cells (ADSCs)‐assisted autologous fat grafting (AFG) significantly improved fat retention in localized scleroderma patients. This pilot study shows the potential advantage of ADSCs‐assisted AFG in the correction of facial atrophy of localized scleroderma patients. SVF, stromal vascular fraction.

Treatment of oral squamous cell carcinoma remains a challenge due to a high incidence of treatment resistance, which is followed by tumor recrudescence and metastasis to the lymph nodes. Thus, it is important to explore novel inhibitors of OSCC. Here, we aimed to identify drugs that may cooperate with histone deacetylase inhibitors to reverse the EMT, inhibit EMT and cell migration and invasion, and contribute to therapeutic efficacy. We found that treatment with 4sc-202 potently reversed the EMT and thereby inhibited cell migration and invasion in vitro, in part by inducing expression of the FoxO1 tumor-suppressor gene. Furthermore, 4sc-202 also synergized with Ink-128 to inhibit tumor migration and invasion in vitro. Mechanistically, 4sc-202 induced FoxO1 expression, whereas Ink-128 promoted nuclear translocation of FoxO1. Our findings indicated that FoxO1 might reverse the EMT by interacting with Twist1 in OSCC. In conclusion, we identified an effective combination therapy involving class I histone deacetylase and mammalian target of rapamycin complex 1/2 inhibition that effectively blocked the EMT of tumor cells by upregulating FoxO1 expression to inhibit Twist1 transcription. These data have implications for developing new targets for early diagnosis and treatment of OSCC.

Localized scleroderma (LoS) is an autoimmune connective tissue disorder leading to serious long-term aesthetic impairment on patients. Objective evaluation methods are badly needed to facilitate the evaluation of the surgical treatment on individual patients and clinical studies. To develop and assess the reliability and validity of Peking Union Medical College LoS facial aesthetic index (PUMC LoSFAI). Twelve experts devoted their time and resources in the development and validation. LoS patients in the stable phase were recruited. Reliability and validity was then assessed. LoS patients were evaluated by two plastic surgeons using PUMC LoSFAI and LoS skin damage index (LoSDI). The PUMC LoSFAI comprises 4 domains for the local assessment (surface area of lesion, dyspigmentation, skin thickness and soft tissue atrophy) and 3 domains for the overall assessment (facial symmetry, proportion and profile) to describe LoS facial aesthetic impairment. Face-Q was completed by patients at each visit. Thirty-two LoS patients had 96 visits, during which 138 lesions were assessed. PUMC LoSFAI and 7 domains demonstrated substantial to excellent inter- and intra-rater reliability (ICC 0.995, κw 0.72-0.91, r 0.85–0.99, respectively). Seven domains considered to be important to extremely important variables (mean rank 3.2–3.8) had high I-CVI (> 0.78) and S-CVI (0.93). PUMC LoSFAI correlated excellently with LoSDI (r = 0.933, P

Purpose: On the basis of the recent discovery of mutations in Bruton tyrosine kinase (BTK) in follicular lymphoma, we studied their functional properties. Experimental Design: We identified novel somatic BTK mutations in 7% of a combined total of 139 follicular lymphoma and 11 transformed follicular lymphoma cases, none of which had received prior treatment with B-cell receptor (BCR) targeted drugs. We reconstituted wild-type (WT) and mutant BTK into various engineered lymphoma cell lines. We measured BCR-induced signal transduction events in engineered cell lines and primary human follicular lymphoma B cells. Results: We uncovered that all BTK mutants destabilized the BTK protein and some created BTK kinase-dead mutants. The phospholipase C gamma 2 (PLCγ2) is a substrate of BTK but the BTK mutants did not alter PLCγ2 phosphorylation. Instead, we discovered that BTK mutants induced an exaggerated AKT phosphorylation phenotype in anti-Ig-treated recombinant lymphoma cell lines. The short hairpin RNA–mediated knockdown of BTK expression in primary human nonmalignant lymph node–derived B cells resulted in strong anti-Ig-induced AKT activation, as did the degradation of BTK protein in cell lines using ibrutinib-based proteolysis targeting chimera. Finally, through analyses of primary human follicular lymphoma B cells carrying WT or mutant BTK, we detected elevated AKT phosphorylation following surface Ig crosslinking in all follicular lymphoma B cells, including all BTK-mutant follicular lymphoma. The augmented AKT phosphorylation following BCR crosslinking could be abrogated by pretreatment with a PI3Kδ inhibitor. Conclusions: Altogether, our data uncover novel unexpected properties of follicular lymphoma–associated BTK mutations with direct implications for targeted therapy development in follicular lymphoma. See related commentary by Afaghani and Taylor, p. 2123

Cell sheets from bone marrow mesenchymal stem cells (BMSC) have been widely used in the field of bone tissue engineering, although their source remains a challenging issue. In this study, adipose-derived stem cells (ADSC) were induced to differentiate into osteoblasts, and the incorporation of coral scaffolds with ADSC sheets for bone formation at an ectopic site was also investigated. First, ADSC isolated from inguinal adipose tissue of New Zealand rabbits were cultured for two weeks without passaging under osteogenic induction, and the microstructures of cell sheets were analysed by histological and scanning electron microscope (EM) observation. In addition, the activity of alkaline phosphatase (ALP) and alizarin red staining was also measured to detect their osteogenic ability. Subsequently, ADSC were proved to be able to proliferate well when seeded on the coral scaffolds. Next, coral scaffolds were wrapped in cell sheets to prepare sheet-coral complexes, which were implanted into subcutaneous pockets in nude mice. At eight weeks after implantation, gross examination, microcomputed tomography (MicroCT), and histological analysis were investigated to assess new bone formation. MicroCT scanning and histological analysis showed that there was more highly dense tissue formed in the complex group than control group (p=0.0004). These results indicated that osteoblastic ADSC sheets could be used to construct engineered bone and the incorporation of coral scaffolds with ADSC sheets significantly improved bone formation, providing a newly approach for bone tissue engineering.

Bioactive scaffolds from synthetical polymers or decellularized cartilage matrices have been widely used in osteochondral regeneration. However, the risks of potential immunological reactions and the inevitable donor morbidity of these scaffolds have limited their practical applications. To address these issues, a biological extracellular matrix (ECM) scaffold derived from allogenic decellularized bone marrow mesenchymal stem cell (BMSC) sheets was established for osteochondral reconstruction. BMSCs were induced to form cell sheets. Three different concentrations of sodium dodecyl sulfate (SDS), namely, 0.5%, 1%, and 3%, were used to decellularize these BMSC sheets to prepare the ECM. Histological and microstructural observations were performed in vitro and then the ECM scaffolds were implanted into osteochondral defects in rabbits to evaluate the repair effect in vivo. Treatment with 0.5% SDS not only efficiently removed BMSCs but also successfully preserved the original structure and bioactive components of the ECM When compared with the 1% and 3% SDS groups, histological observations substantiated the superior repair effect of osteochondral defects, including the simultaneous regeneration of well-vascularized subchondral bone and avascular articular cartilage integrated with native tissues in the 0.5% SDS group. Moreover, RT-PCR indicated that ECM scaffolds could promote the osteogenic differentiation potential of BMSCs under osteogenic conditions while increasing the chondrogenic differentiation potential of BMSCs under chondrogenic conditions. Allogenic BMSC sheets decellularized with 0.5% SDS treatment increased the recruitment of BMSCs and significantly improved the regeneration of osteochondral defects in rabbits, thus providing a prospective approach for both articular cartilage and subchondral bone reconstruction with cell-free transplantation.

Background: Although the effectiveness of buprenorphine-naloxone (BUP-NX) has been established, real-world evidence on the benefits of early treatment initiation is limited. Objective: To evaluate the association between early BUP-NX initiation and health-related outcomes among insured adults with opioid use disorder (OUD). Methods: We conducted a cross-sectional analysis using the Optum’s de-identified Clinformatics® Data Mart Database from 2010 to 2018. Patients who initiated BUP-NX within 30 days of OUD diagnosis were classified as early initiators. Patients who initiated BUP-NX later, but within the one-year follow-up, were defined as late initiators. Outcomes included opioid overdose, opioid overdose-related emergency department (ED) visits, and all-cause healthcare cost during the year after OUD diagnosis. We employed generalized linear models to compare outcomes between early and late initiators, adjusting for baseline covariates and accounting for missing information for covariates using multiple imputation. Results: A total of 8,388 patients with OUD were identified; mean age was 39.9 years; 36% were female; and 67.6% were early initiators. Early initiators had an estimated 42% lower rate of opioid overdose (adjusted rate ratio (aRR) = 0.58; 95% confidence interval (CI): 0.52, 0.64); 51% lower rate of opioid overdose-related ED visits (aRR = 0.49; 95% CI: 0.44, 0.55); and 31% lower total healthcare cost (adjusted cost ratio = 0.69; 95% CI: 0.66, 0.72), compared to late initiators. Conclusion: Compared to late BUP-NX initiation, early initiation was associated with a lower risk of opioid overdose and opioid overdose-related ED visits, and reduced total healthcare cost among insured adult patients with OUD.

Background Osteosarcoma (OS) is a highly prevalent bone malignancy with poor clinical outcomes. Expression of the circular RNA, hsa_circ_0078767 (circFAM120B) is elevated in OS, however, its mechanisms in OS are unclear. Methods CircFAM120B levels were detected in OS tissue and cell lines. Silenced circFAM120B experiments were performed to assess its effects on OS in vitro cancer phenotypes and in vivo tumor growth. Then, bioinformatics analyses were used to predict circFAM120B target microRNAs (miRNAs) and associated genes. Results CircFAM120B and the transcription factor, PTBP1 were elevated in OS tissue and cell lines, while miR-1205 was poorly expressed. Silenced circFAM120B significantly suppressed in vitro OS cell proliferation and invasion, and inhibited in vivo tumor growth. CircFAM120B also appeared to function as an miR-1205 sponge, as miR-1205 bound to PTBP1. Interestingly, overexpressed PTBP1 (or miR-1205 inhibition) reversed the inhibitory effects mediated by circFAM120B downregulation in OS cells. Conclusion We hypothesize circFAM120B functions as a miR-1205 sponge to elevate PTBP1 levels, enhancing OS progression and associated malignant phenotypes. Thus, circFAM120B may function as a crucial mediator during OS progression.

The BCL2-inhibitor, Venetoclax (VEN), has shown significant anti-leukemic efficacy in combination with the DNMT-inhibitor, Azacytidine (AZA). To explore the mechanisms underlying the selective sensitivity of mutant leukemia cells to VEN and AZA, we used cell-based isogenic models containing a common leukemia-associated mutation in the epigenetic regulator ASXL1. KBM5 cells with CRISPR/Cas9-mediated correction of the ASXL1G710X mutation showed reduced leukemic growth, increased myeloid differentiation, and decreased HOXA and BCL2 gene expression in vitro compared to uncorrected KBM5 cells. Increased expression of the anti-apoptotic gene, BCL2, was also observed in bone marrow CD34+ cells from ASXL1 mutant MDS patients compared to CD34+ cells from wild-type MDS cases. ATAC-sequencing demonstrated open chromatin at the BCL2 promoter in the ASXL1 mutant KBM5 cells. BH3 profiling demonstrated increased dependence of mutant cells on BCL2. Upon treatment with VEN, mutant cells demonstrated increased growth inhibition. In addition, genome-wide methylome analysis of primary MDS samples and isogenic cell lines demonstrated increased gene-body methylation in ASXL1 mutant cells, with consequently increased sensitivity to AZA. These data mechanistically link the common leukemia-associated mutation ASXL1 to enhanced sensitivity to VEN and AZA via epigenetic upregulation of BCL2 expression and widespread alterations in DNA methylation.

The morbidity and mortality of lung cancer are increasing. The Corona Virus Disease 2019 (COVID-19) is caused by novel coronavirus 2019-nCoV-2, leading to subsequent pulmonary interstitial fibrosis with chronic inflammatory changes, e.g., inflammatory factors repeatedly continuously stimulating and attacking the alveolar epithelial cells. Meanwhile, 2019-nCoV-2 can activate PI3K/Akt and ERK signaling pathways, which can play the double roles as both anti-inflammatory and carcinogenic factors. Moreover, hypoxemia may be developed, resulting in the up-regulation of HIF-1 α expression, which can be involved in the occurrence, angiogenesis, invasion and metastasis of lung cancer. Additionally, the immune system in 2019-nCoV-2 infected cases can be suppressed to cause tumor immune evasion. Therefore, we speculate that COVID-19 may be a risk factor of secondary lung cancer.

Spore-forming probiotic Bacillus spp. have received extensively increasing scientific and commercial interest, but raised the concerns in the potential risks and pathogenesis. In this study, 50 commercial probiotic products were collected from all over the country and Bacillus spp. isolated from products were evaluated for the safety on the aspects of hemolytic activity, contamination profiles, toxin genes, cytotoxicity, antimicrobial resistance, and genotyping. 34 probiotic products (68%) exhibited hemolysis, including 19 human probiotics, 9 animal probiotics, and 6 plant probiotics. 28 products (56%) contained other bacteria not labeled in the ingredients. 48 strains in Bacillus spp. including 17 B. subtilis group isolates, 28 B. cereus, and 3 other Bacillus spp. were isolated from human, food animal, and plant probiotic products. Detection rates of enterotoxin genes, nheABC and hblCDA, and cytotoxin cytK2 in 48 Bacillus spp. isolates were 58%, 31%, and 46%, respectively. Also, one isolate B. cereus 34b from an animal probiotic product was positive for ces, encoding cereulide. 28 of 48 Bacillus spp. isolates were cytotoxic. 19 of 28 B. cereus isolates maintained to exhibit hemolysis after heat treatment. All 48 Bacillus spp. isolates exhibited resistance to lincomycin, and 5 were resistant to tetracycline. The genotyping of commercial probiotic Bacillus spp. reported in this study showed that ces existed in B. cereus 34b with the specific sequence type (ST1066). These findings support the hypothesis that probiotic products were frequently contaminated and that some commercial probiotics consisted of Bacillus spp. may possess toxicity and antimicrobial resistance genes. Thus, the further efforts are needed in regarding the surveillance of virulence factors, toxins, and antibiotic resistance determinants in probiotic Bacillus spp.

BACKGROUND No reports of the potential areas of surgeons' aesthetics in blepharoplasty. AIMS To explore the association between the surgeons' own double eyelid morphology and their aesthetics and surgical outcome. METHODS An investigation of 1605 patients was carried collecting the data of double eyelid shape, based on which to propose a preliminary double eyelid classification for analyzing the aesthetics of surgeons. Ten double eyelid surgical cases were randomly collected from each surgeon according to the inclusion criteria, whose double eyelid shape, ideal shape, the selection tendency of surgical approach, most cared factor during surgery, and design concept of eyelid shape were collected. Structural equation modeling (SEM) was performed to assess the association between participants' double eyelid shape, aesthetics, and blepharoplasty outcome. RESULTS Fifty-three double eyelid surgeons were enrolled for study, whose double eyelids shapes mostly were obviously fan-shaped (37.74%) and low-parallel eyelid (26.42%), and the ideal shapes were obviously fan-shaped (41.51%) and high-parallel eyelid (24.53%). 54.72% of the subjects preferred to use the full-incisional method. 45.28% cared for long-term outcome most. Regarding blepharoplasty outcome style, 49.06% of the subjects preferred obviously fan-shaped type and 24.53% for high-parallel type. SEM showed that aesthetics rank increased by 0.692 points for surgeons' shape (P

The Rx (prescription) for Addiction and Medication Safety (RAMS) program was developed during the 2017 through 2018 academic year to educate students from 6 selected Rhode Island public high schools about opioid misuse, overdose, and recovery. During 2016, 3 schools participated in the RAMS program and returned for RAMS-PEER in 2017; 3 schools were newly recruited in 2016. Tenth graders returned from schools that participated during RAMS in 2016, and all ninth graders were new. Our study's aim was to evaluate the overall effect and spillover benefit of the RAMS-PEER intervention from tenth to ninth graders by surveying students both before and after the education program. Survey questions were modified from the 2015 Youth Risk Behavior Survey and the 2015 Ontario Study Survey. Student responses were matched for preintervention and postintervention analysis using a unique identifier. We observed an improvement in knowledge of opioid misuse; however, we found no evidence of a significant spillover benefit.

Low sensitivity to chemotherapy has been a major challenge in the treatment of non-small-cell lung cancer (NSCLC). It is of great clinical significance to discover its mechanisms to improve cell sensitivity to chemotherapeutic drugs. The forkhead box subfamily O (FOXO) transcriptional factors are downstream factors of the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway and are reported to play pro-apoptotic roles in a variety of cells including NSCLC cells. But their roles and mechanisms in mediating cell response to chemotherapy remain to be discovered. We proposed that FOXO1 and FOXO3a may increase the sensitivity of NSCLC cells to cisplatin. Moreover, we presumed that LY294002, an inhibitor of the PI3K/AKT pathway, may enhance the cytotoxic effects of cisplatin through upregulating FOXO1 and FOXO3a. In the present study, we found that cisplatin initially increased the expressions and nuclear accumulation of FOXO1 and FOXO3a in NSCLC. Knockdown of FOXO1 and FOXO3a significantly decreased the cell sensitivity to cisplatin in vitro and in vivo. Moreover, inhibition of FOXO1 and FOXO3a attenuated cisplatin-induced cell apoptosis independent of Bim, a pro-apoptotic protein downstream of the FOXOs. Moreover, LY294002 synergistically increased the cytotoxic effects of cisplatin. Mechanistically, LY294002 increased the expressions and nuclear accumulation of FOXO1 and FOXO3a. Knockdown of FOXO1 and FOXO3a abrogated the enhancing effect of LY294002 on cisplatin. Taken together, our results suggested that FOXO1 and FOXO3a sensitize NSCLC cells to cisplatin and mediate the enhancing effects of LY294002 on cisplatin.

Cancer is a serious threat to human health. With the increasing resistance to known drugs, it is still urgent to find new drugs or pro-drugs with anti-tumor effects. Natural products produced by microorganisms have played an important role in the history of drug discovery, particularly in the anticancer and anti-infective areas. The plant rhizosphere ecosystem is a rich resource for the discovery of actinomycetes with potential applications in pharmaceutical science, especially Streptomyces. We screened Streptomyces-like strains from the rhizosphere soil of wheat (Triticum aestivum L.) in Hebei province, China, and thirty-nine strains were obtained. Among them, the extracts of 14 isolates inhibited the growth of colon tumor cell line HCT-116. Strain NEAU-wh-3-1 exhibited better inhibitory activity, and its active ingredients were further studied. Then, 16S rRNA gene sequence similarity studies showed that strain NEAU-wh3-1 with high sequence similarities to Embleya scabrispora DSM 41855T (99.65%), Embleya hyalina MB891-A1T (99.45%), and Streptomyces lasii 5H-CA11T (98.62%). Moreover, multilocus sequence analysis based on the five other house-keeping genes (atpD, gyrB, rpoB, recA, and trpB) and polyphasic taxonomic approach comprising chemotaxonomic, phylogenetic, morphological, and physiological characterization indicated that the isolate should be assigned to the genus Embleya and was different from its closely related strains, therefore, it is proposed that strain NEAU-wh3-1 may be classified as representatives of a novel species of the genus Embleya. Furthermore, active substances in the fermentation broth of strain NEAU-wh-3-1 were isolated by bioassay-guided analysis and identified by nuclear magnetic resonance (NMR) and mass spectrometry (MS) analyses. Consequently, one new Zincophorin analogue together with seven known compounds was detected. The new compound showed highest antitumor activity against three human cell lines with the 50% inhibition (IC50) values of 8.8&ndash, 11.6 &mu, g/mL and good antibacterial activity against four pathogenic bacteria, the other known compounds also exhibit certain biological activity.

Cell sheet techniques are widely used in bone engineering. However, vascularization remains a challenge in fabricating vascularized engineered bone. The goal of this study was to induce adipose-derived stem cell (ADSC) osteogenic and angiogenic lineage differentiation and investigate the use of bidiretionally differentiated ADSCs for bone regeneration. ADSCs were cultured to form an osteogenic cell sheet. Other ADSCs were induced to differentiate into endothelial progenitor cells (EPCs), which were identified and characterized by morphological observation and CD31 immunofluorescent staining. Then, the ADSC sheet-EPC complexes were implanted subcutaneously into nude mice, while ADSC sheets alone were implanted as a control. After 8 weeks of transplantation, microcomputed tomography (micro-CT) and histological observation were used to assess bone formation. We then implanted the complexes in calvarial defects in rabbits and assessed bone repair by micro-CT and histological analysis. The ADSC sheets consisted of multiple layers of cells and extracellular matrix. The obtained EPCs formed capillary-like structures and expressed the specific antigen marker CD31. The osteogenic ADSC sheet-EPC complexes formed dense and well-vascularized new bone tissue at 8 weeks after implantation. Bone density was significantly lower in the control group than in the complex group (p

Botulinum toxin injection, micro-needling injection, and filler injection are the most widely used non-surgical facial cosmetic treatments. Hypersensitivity reactions associated with injections have not received sufficient attention due to their low incidence rates. The authors tried to summarize the characteristics of the hypersensitivity caused by cosmetic injections and helped to improve the diagnosis and treatment. A comprehensive search of the PubMed database to September 2019 was performed. Articles were screened using predetermined inclusion and exclusion criteria. Data collected included patient characteristics, injection information (injected material, trade name, location of injection, symptoms of allergy, time of onset), diagnostic examination, treatment, and prognosis. A total of 14 articles (57 patients) were included. There were 3 patients receiving botulinum toxin injection, 3 patients receiving micro-needling injection, and 46 patients receiving hyaluronic acid injection. Five patients were injected with collagen, polyacrylamide, paraffin, alkyl-imide or hyaluronidase, separately. The symptoms were redness, swelling, itching or induration at the injection site. The diagnosis and treatment methods vary greatly depending on the injection fillers. Removing the injected filler is the primary treatment. Although hypersensitivity reaction is a relatively uncommon adverse event, more practice guidelines and research on diagnosis and treatment are demanded to help improve the outcomes. This journal requires that authors assign a level of evidence to each article. For a full description of these evidence-based medicine ratings, please refer to the table of contents or the online instructions to authors www.springer.com/00266 .

There are a number of patients who develop severe anxiety or depression after receiving facial cosmetic injections. We presented a 32-year-old woman who developed frequent panic, tension headache, tachycardia, shortness of breath, and sleep disorder for a year after the injection of hyaluronic acid on her forehead and glabella. Brain magnetic resonance imaging (MRI) showed multiple flaky and slightly longer T1, slightly longer T2 signals on the anterior frontal white matter. However, the patient's brain MRI scan 5 months before the injection showed no such performance. The patient was asked to consult the psychiatrist and was diagnosed with anxiety disorder. We name such phenomenon by Emotional disorder syndrome after cosmetic facial injection and assume that there are three major reasons for the emergence of this syndrome. One reason may be that emotional disorder is caused by the mental state of the patient. The second reason to explain the emotional disorder might be the frontal lobe syndrome caused by the frontal embolism during the filler injection. Another reason may be leukoaraiosis, a brain white matter change which may cause depression and anxiety.

Introduction: Individual mutations in the SCN5A-encoding cardiac sodium channel α-subunit usually cause a single cardiac arrhythmia disorder, some cause mixed biophysical or clinical phenotypes. Here we report an infant, female patient harboring a N406K mutation in SCN5A with a marked and mixed biophysical phenotype and assess pathogenic mechanisms. Methods and Results: A patient suffered from recurrent seizures during sleep and torsades de pointes with a QTc of 530 ms. Mutational analysis identified a N406K mutation in SCN5A. The mutation was engineered by site-directed mutagenesis and heterologously expressed in HEK293 cells. After 48 hours incubation with and without mexiletine, macroscopic voltage-gated sodium current (INa) was measured with standard whole-cell patch clamp techniques. SCN5A-N406K elicited both a significantly decreased peak INa and a significantly increased late INa compared to wide-type (WT) channels. Furthermore, mexiletine both restored the decreased peak INa of the mutant channel and inhibited the increased late INa of the mutant channel. Conclusion: SCN5A-N406K channel displays both “gain-of-function” in late INa and “loss-of-function” in peak INa density contributing to a mixed biophysical phenotype. Moreover, our finding may provide the first example that mexiletine exerts a dual rescue of both “gain-of-function” and “loss-of-function” of the mutant sodium channel.

The hemispherical phased transducer maximizes the coverage of the skull and the ultrasonic energy per unit area of the skull is minimized, thereby reducing the risk of skull burns, but the transducer has a small focal area adjustment range, increasing the focal length of treatment is an urgent question for this type of transducer. In this paper, a three-dimensional high-intensity focused ultrasound (HIFU) transcranial propagation model is established based on the human head structure. The finite difference time domain (FDTD) is combined with the Westervelt acoustic wave nonlinear propagation equation and Penne's biological heat conduction equation for numerical simulation of the sound pressure field and temperature field. Forming a treatable focal area in a small-opening hemispherical transducer with a small amount of numerical simulation calculation focusing at a set position to determine the minimum partial excitation area ratio of focusing. And then, applying these preliminary results to a large-opening diameter hemispherical transducer and the temperature field formed by it or full excitation is studied. The results show that the focus area with the excitation area ratio of less than 22% moves forward to the transducer side when the excitation sound is formed. When the excitation area ratio is greater than or equal to 23%, it focuses at the set position. In the case of partial incentives, using 23% of the partial array, the adjustable range of the treatable focal area formed in the three-dimensional space is larger than that of the full excitation.

// Zheng Ge 1, 2 , Xilian Zhou 3 , Yan Gu 3 , Qi Han 3 , Jianyong Li 3 , Baoan Chen 1, 2 , Qinyu Ge 4 , Elanora Dovat 5 , Jonathon L. Payne 5, 6 , Tianyu Sun 7 , Chunhua Song 2, 5 , Sinisa Dovat 2, 5 1 Department of Hematology, Zhongda Hospital, Medical School of Southeast University, Nanjing 210009, China 2 International Cooperative Leukemia Group and International Cooperative Laboratory of Hematology, Zhongda Hospital, Medical School of Southeast University, Nanjing 210009, China 3 Department of Hematology, The First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing 210029, China 4 State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China 5 Department of Pediatrics, Pennsylvania State University Medical College, Hershey, PA 17033, USA 6 Loma Linda University School of Medicine, Department of Basic Sciences, Loma Linda, CA 92350, USA 7 Department of Internal Medicine, Lewis Katz School of Medicine at Temple University, Philadelphia, PA 19140, USA Correspondence to: Zheng Ge, email: Janege879@hotmail.com Sinisa Dovat, email: sdovat@hmc.psu.edu Keywords: BCL6, BACH2, IKZF1, acute lymphoblastic leukemia Received: October 11, 2016 Accepted: November 24, 2016 Published: December 20, 2016 ABSTRACT B-Cell CLL/Lymphoma 6 ( BCL6 ) is a proto-oncogene that is highly expressed in acute lymphoblastic leukemia (ALL). BTB and CNC Homology 1 Basic Leucine Zipper Transcription Factor 2 (BACH2) is a suppressor of transcription. The BACH2 – BCL6 balance controls selection at the pre-B cell receptor checkpoint by regulating p53 expression. However, the underlying mechanism and the clinical relevance of the BCL6/BACH2 axis are unknown. Here, we found that Ikaros, a tumor suppressor encoded by IKZF1 , directly binds to both the BCL6 and BACH2 promoters where it suppresses BCL6 and promotes BACH2 expression in B-cell ALL (B-ALL) cells. Casein kinase 2 (CK2) inhibitors increase Ikaros function thereby inhibiting BCL6 and promoting BACH2 expression in an Ikaros-dependent manner. We also found that the expression of BCL6 is higher while BACH2 expression is lower in patients with B-ALL than normal bone marrow control. High BCL6 and low BACH2 expression is associated with high leukemic cell proliferation, unfavorable clinical and laboratory features, and inferior outcomes. Moreover, IKZF1 deletion is associated with high BCL6 and low BACH2 expression in B-ALL patients. CK2 inhibitors increase Ikaros binding to the promoter of BCL6 and BACH2 and suppress BCL6 while promoting BACH2 expression in the primary B-ALL cells. Our data indicates that Ikaros regulates expression of the BCL6/BACH2 axis in B-ALL. High BCL6 and low BACH2 expression are associated with Ikaros dysregulation and have a potential effect on the development of B-ALL.

Objectives The goal of this study was to prepare lyophilized platelet-rich fibrin (L-PRF) and analyze the combined use of L-PRF and osteogenic bone marrow mesenchymal stem cell (BMSC) sheet fragments for bone tissue engineering via in vivo injection. Methods First, fresh PRF (F-PRF) was lyophilized to prepare L-PRF, the characteristics of which were examined through gross morphological, and histological and microstructural observations. In addition, the kinetics of growth factor release from L-PRF and F-PRF were also determined by enzyme-linked immunosorbent assay (ELISA). Subsequently, after assessing the proliferation and osteogenic differentiation of BMSCs exposed to L-PRF or F-PRF in vitro, we subcutaneously injected BMSC sheet fragments with L-PRF or F-PRF into nude mice and assessed bone formation through microcomputed tomography and histological analyses. Results We observed that L-PRF released growth factors that favored BMSC proliferation and osteogenic differentiation in vitro. The combined use of L-PRF and osteogenic BMSC sheet fragments enabled bone tissue regeneration in vivo, and no significant difference between the F-PRF and L-PRF groups was observed (P = 0.24). Conclusions The results of this study demonstrate that the combined use of L-PRF and osteogenic BMSC sheets may have potential in the fabrication of engineered bone., Bioengineering; Cell biology; Materials science; Molecular biology; Stem cell research; lyophilized platelet-rich fibrin; bone marrow mesenchymal stem cells; cell sheet; bone tissue engineering.

Most nanoparticles (NPs) are reported to block autophagic flux, accompanied by accumulated p62/SQSTM1 resulting from degradation inhibition. p62 also acts as a multifunctional scaffold protein that contains multiple domains, involved in various cellular processes. However, the autophagy substrate-independent role and regulation at a transcriptional level of p62 upon NPs exposure are ignored. Here, we exposed BEAS-2b cells to silica nanoparticles (SiNPs), and found that p62 degradation was inhibited due to autophagic flux blockade. Mechanically, SiNPs blocked autophagy flux through lysosomal capacity impairment rather than defective autophagosome fusion with lysosomes. Moreover, SiNPs stimulated translocation of NF-E2-related factor 2 (Nrf2) to the nucleus from the cytoplasm, and upregulated p62 transcriptional activation through direct binding of Nrf2 to p62 promoter. Nrf2 siRNA dramatically decreased both mRNA and protein levels of p62. Above two mechanisms led to p62 protein accumulation, therefore increasing IL-1 and IL-6 expression. SiNPs activated nuclear Factor kappa B (NF-κB), which can be alleviated by p62 knockdown. In summary, SiNPs accumulated p62 by both pre- and post-translational mechanisms, resulting in pulmonary inflammation. These findings improve our understanding of SiNP-induced pulmonary damage and molecular targets to antagonise it.

The GRAS (generally recognized as safe) status of Enterococcus has not yet been authenticated, but enterococci, as probiotics, have been increasingly applied in human healthcare and animal husbandry, for instance as a dietary supplement, feed additive, or growth promotor. The food chain is the important route for introducing enterococci into the human gut. The pathogenicity of Enterococcus from probiotic products requires investigation. In the study, 110 commercial probiotic products used for human, animal, aquaculture, and plants were examined, among which 36 enterococci were identified, including 31 from Enterococcus faecium, 2 from E. faecalis, 2 from E. casseliflavus, and 1 from E. gallinarum. Strikingly, 28 of the 36 enterococci isolated from probiotics here did not mention the presence of Enterococcus in the labeled ingredients, and no Enterococcus isolates were found from 5 animal probiotics that were labeled with the genus. In total, 35 of the 110 products exhibited hemolysis, including 5 (10.6%) human probiotics, 14 (41.2%) animal probiotics, 8 (57.1%) aquaculture probiotics, and 8 (53.3%) plant probiotics. The detection rates of virulence factors associated with adhesion, antiphagocytosis, exoenzyme, biofilm, and other putative virulence markers (PVM) in 36 enterococci were 94.4%, 91.7%, 5.6%, 94.4% and 8.3%. Twenty-six of the 36 isolated strains exhibited biofilm formation ability, where 25 strains (69.4%) and one (2.8%) were strong and weak biofilm producers, respectively. We analyzed the resistance rates against erythromycin (97%), vancomycin and ciprofloxacin (8%), tetracycline (3%), and high-level aminoglycosides (0%), respectively. High detection rates of msrC/lsaA (86%) and aac(6′)-Ii (86%) were observed, followed by vanC (8%), tetM (3%). The Tn5801-tetM-like integrative conjugative element (ICE) was identified in E. gallinarum, exhibiting resistance to tetracycline (64 μg/mL). Seven probiotic E. faecalis and E. faecium, as active ingredients in human probiotics, shared the same STs (sequence types) and were distinct from the STs of other contaminated or mislabeled enterococci, indicating that two particular STs belonged to native probiotic isolates. These findings advocate appropriate assessments of enterococci when used in probiotics.

The assessment of single gene such as ERCC1, TYMS, RRM1, TUBB3, EGFR, KRAS, BRAF, PIK3CA, ALK, and ROS1 is now widely applied in therapeutic decisions of non-small cell lung cancer (NSCLC). The aim of our study was to concurrently analyze these genes and evaluate their clinical significance in patients with NSCLC. Rearrangement of ALK and ROS1 was analyzed in 120 patients using FISH assays. Somatic mutation of EGFR, KRAS, BRAF, PIK3CA and mRNA expression of ERCC1, TYMS, RRM1, TUBB3, EGFR were examined by liquidchip platform in 350 patients . Data on clinical features were obtained from medical records of 119 patients, and the follow-up was conducted in 106 patients who received platinum-based adjuvant chemotherapy. We identified 5.0 % ALK rearrangements, 1.7 % ROS1 rearrangements, 36.6 % EGFR mutations, 8.9 % KRAS mutations, 0 % BRAF mutations, and 4.0 % PIK3CA mutations. Double or coexisting mutations were identified in 13 patients. Significant correlations were observed among EGFR, KRAS mutation, ERCC1, TYMS, RRM1, TUBB3, EGFR expression, and clinical features, especially histology (P

Introduction Hyaluronic acid injections is relatively safe with little risk of complications. Although herpes reactivation after the injection of hyaluronic acid is rare, it produces quite a huge pressure and panic on patients. Quite a lot cosmetic practitioners have no awareness of preventing, diagnosing, and giving correct treatment in time due to lack of experience. Patient concerns A 24-year-old woman presented with erythema, crusted papules, pain and swelling on the nose after receiving the injection of hyaluronic acid. A swab of the discharge fluid was obtained for bacterial and viral culture, showing positive for herpes simplex virus. Diagnosis The patient was diagnosed as herpes reactivation after the injection of hyaluronic acid. Interventions The patient underwent antiviral therapy with acyclovir 400 mg, 3 times daily for seven days. Outcomes After a week of antiviral treatment, the clinical signs improved. Conclusion Herpes reactivation after the injection of hyaluronic acid is quite rare but needed sufficient attention of cosmetic practitioners to make the proper diagnosis, prevention and treatment.

The hierarchical interactions between the dental epithelium and dental mesenchyme represent a common paradigm for organogenesis. During tooth development, various morphogens interact with extracellular components in the extracellular matrix and on the cell surfaces to transmit regulatory signaling into cells. We recently found pivotal roles of FAM20B-catalyzed proteoglycans in the control of murine tooth number at embryonic stages. However, the expression pattern of proteoglycans in embryonic teeth has not been well understood. We extracted total RNA from E14.5 murine tooth germs for semi-quantitative RT-PCR analysis of 29 proteoglycans, and identified 23 of them in the embryonic teeth. As a major subfamily of FAM20B-catalyzed proteoglycans, Syndecans are important candidates being potentially involved in the tooth development of mice. We examined the expression pattern of Syndecans in embryonic teeth using in situ hybridization (ISH) and immunohistochemistry (IHC) approaches. Syndecan-1 is mainly present in the dental mesenchyme at early embryonic stages. Subsequently, its expression expands to both dental epithelium and dental mesenchyme. Syndecan-2 is strongly expressed in the dental mesenchyme at early embryonic stages, then shifts to the stratum intermedium and inner dental epithelium at cap stages. Syndecan-3 shows a gradually increased expression that initially in the dental epithelium of both incisors and molars and then in the inner dental epithelium and stratum intermedium in molars alone. Syndecan-4 is localized in the dental epithelium in incisors and the dental follicle mesenchyme in molars at early cap stage. The spatiotemporal expression pattern of Syndecans in murine embryonic teeth suggest potential roles of these proteoglycans in murine tooth morphogenesis.

The primary objective of this study was to mask bitter taste and decrease the disintegration time of carbinoxamine maleate (CAM) orally disintegrating tablets (ODTs). In order to screen the prescription of ODTs, a novel modified in vitro disintegration method (MIVDM) was developed to measure the in vitro disintegration time. In this method, different concentrations of ethanol served as disintegration medium in order to delay the in vitro water absorption and disintegration process of tablets. The MIVDM demonstrated good in vitro and in vivo correlation and proved more precise and discriminative than other reported methods. In this research, ion exchange resins (IERs) were used to mask bitter taste for improving mouthfeel. The drug-resin ratio and reaction temperature were investigated to obtain the optimum carbinoxamine resin complexes (CRCs). The characterization of CRCs revealed an amorphous state. ODTs were prepared by direct compression. Superdisintegrants and diluents of ODTs were screened first. Further optimization was carried out by using Box-Behnken design. The effect of (X1) mannitol/microcrystalline cellulose ratio, (X2) the amount of low-substituted hydroxypropylcellulose and (X3) the hardness was investigated for achieving the lowest (Y) in vitro disintegration time. Technological characterization, wetting time, water absorption ratio, and roughness degree were evaluated. The CRCs and ODTs proved successful taste-masking efficiency. The end product improved patients' compliance. The developed MIVDM was practical for commercial use.

Microbial infections continually present a major worldwide public healthcare threat, particularly in instances of impaired wound healing and biomedical implant fouling. The development of new materials with the desired antimicrobial property to avoid and treat wound infection is urgently needed in wound care management. This study reports a novel dual-functional biodegradable dextran-poly(ethylene glycol) (PEG) hydrogel covalently conjugated with antibacterial Polymyxin B and Vancomycin (Vanco). The hydrogel is designed as a specialized wound dressing that eradicates existing bacteria and inhibits further bacteria growth, while, ameliorating the side effects of antibiotics and accelerating tissue repair and regeneration. The hydrogel exhibits potent antibacterial activities against both gram-negative bacteria Escherichia coli (E. coli) and gram-positive bacteria Staphylococcus aureus (S. aureus) with no observable toxicity to mouse fibroblast cell line NIH 3T3. These results demonstrate the immense potential of dextran-PEG hydrogel as a wound dressing healthcare material in efficiently controlling bacteria growth in complex biological systems.

// Bo Deng 1, * , Tianyu Sun 1, * , Bo Tang 1, * , Shaolin Tao 1 , Poming Kang 1 , Kai Qian 1 , Bin Jiang 1 , Kun Li 1 , Kunkun Li 1 , Jinghai Zhou 1 , Ruwen Wang 1 and Qunyou Tan 1 1 Department of Thoracic Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, China * These authors have contributed equally to this work Correspondence to: Qunyou Tan, email: tanqy001@163.com Keywords: adenoviral p53 gene; non-small cell lung cancer; radical surgery; recurrence; metastasis Received: June 27, 2017 Accepted: September 20, 2017 Published: November 06, 2017 ABSTRACT Objective: To assess the efficacy of radical surgery combined with recombinant adenoviral human p53 (rAd-p53) gene therapy in treatment of resectable non-small cell lung cancer. Method: A total of 163 patients with resectable NSCLC meeting the inclusion criteria were randomly assigned to two groups: radical surgery alone (S) and radical surgery plus surgical wound surface injection of 2 x 10 12 rAd-p53 units (SP). All patients were followed up for at least 3 years for efficacy and safety. Study endpoints were loco-regional recurrence or distant metastasis (Rec-Met) rate as primary endpoints, and progression free survival (PFS), overall survival (OS) and safety assessments as secondary endpoints. Results: Recurrence or metastasis (Rec/Met) after surgery were 24/82 (29.27%) in SP group and 37/81 (45.68%) in S group. The difference in the Rec/Met rate was statistically significant (p = 0.0304) by chi-square test. The hazard ratios after adjusting of age and disease stage (S vs. SP) of PFS and OS are 1.772 (95% CI, 1.102 to 2.848) and 2.047 (95% CI, 1.109 to 3.377), respectively. The 3 years PFS and OS for SP vs. S were 71.9% vs. 46.9%, and 88.4% vs. 67.0%, respectively. Differences in PFS and OS between two treatment groups were significant with the p values of 0.0165 and 0.0191, respectively, using Log-Rank test. Conclusions: The wound surface injection of rAd-p53 showed efficacious effects in preventing recurrence or metastasis and improving PFS and OS after a radical surgery in patients with NSCLC.

Dynamin-2 (DNM2) is a GTPase essential for intracellular vesicle formation and trafficking, cytokinesis and receptor endocytosis. Mutations in DNM2 are common in early T-cell precursor acute lymphoblastic leukemia. However, DNM2 expression in other types of ALL are not reported. We studied DNM2 mRNA level in adults with B- and T-cell ALL. We found DNM2 is more highly expressed compared with normals in both forms of ALL. High DNM2 expression is associated with some clinical and laboratory features, inferior outcomes and with leukaemia cell proliferation. We also found Ikaros directly binds the DNM2 promoter and suppresses DNM2 expression. Consequently IKZF1 deletion is associated with high DNM2 expression. Conversely, casein kinase-2 (CK2)-inhibitor increases Ikaros function thereby inhibiting DNM2 expression. Inhibiting DNM2 suppresses proliferation of leukemia cells and synergizes with CK2 inhibition. Our data indicate high DNM2 expression is associated with Ikaros dysregulation and may be important in the development of B-ALL.

INTRODUCTION: Hypoxia-induced apoptosis and arrhythmia are the important cause of sudden infant death syndrome (SIDS). ATP-sensitive K+ (KATP) channels are known to provide a functional linkage between the electrical activity of cell membrane and metabolism. The KCNJ8-encoded Kir6.1 KATP channel critically regulates vascular tone and cardiac adaptive response to systemic metabolic stressors, including sepsis. Previously, we reported two KCNJ8 mutations (E332del, V346I) in a large SIDS cohort that exhibited a marked loss-of-function phenotype, reduction of cell surface expression and increased cell death. Here we further investigate the effect of the mutants on apoptosis.METHODS AND RESULTS: HEK293 cells transfected with IRES-GFP constructs containing Kir6.1-WT or mutant (Kir6.1-E332del or Kir6.1-V346I) with SUR2A in a ratio of 1:1 for 48 hrs. For apoptosis assays, cells were stained with PE-Annexin V and 7-AAD, the apoptotic cells were measured by Flow-cytometry within gated GFP (+) cells. Caspase-3/7 activity was measured with Apo-ONE Homogenous caspase 3/7 assay kit. It showed the apoptotic ratio was significantly increased for mutants Kir6-1-E332del and Kir6.1-V346I compared to Kir6.1-WT. The Caspase-3/7 activity was also increased 1.5∼2.2 folds for Kir6.1-E332del and Kir6.1-V346I over Kir6.1-WT. Furthermore, the mitochondrial superoxide levels was measured by using MitoSOX Red and the basal level of superoxide were significantly higher in mutant Kir6.1-E332del and Kir6.1-V346I compared to Kir6.1-WT (p < 0.05). The mitochondrial membrane potential of Kir6.1-E332del, Kir6.1-V346I and Kir6.1-WT transfected cells were also measured by JC-1 staining with flow cytometry analysis. The results showed that the green fluorescent signal in the mutant cells were increased 1.5 ∼2.5 folds over Kir6.1-WT, indicating the mutants induced mitochondrial membrane damage.CONCLUSIONS: The loss-of-functional Kir6.1 KATP channel mutations found in SIDS cohort induce cell apoptosis through ROS production and mitochondrial dysfunction in heterologous expression system.

INTRODUCTION: Hypoxia-induced apoptosis and arrhythmia are the important cause of sudden infant death syndrome (SIDS). ATP-sensitive K+ (KATP) channels are known to provide a functional linkage between the electrical activity of cell membrane and metabolism. KCNJ8-encoded Kir6.1 KATP channel critically regulates vascular tone and cardiac adaptive response to systemic metabolic stressors, including sepsis. Previously, we identified two KCNJ8 mutations (E332del and V346I) in a large SIDS cohort that exhibited a marked loss-of-function phenotype and reduction of cell surface expression. Here we further investigate the mechanisms underlying the loss-of-functional Kir6.1 KATP channel mutations in SIDS.Methods and Results: A hemagglutinin (HA) epitope was inserted in an extracellular loop of Kir6.1 wild type (WT), Kir6.1-E332del and Kir6.1-V346I channels. HEK293 cells were co-transfected with cDNA encoding HA-tagged Kir6.1-WT, HA-tagged Kir6.1-E332del or -V346I and SUR2A in a ratio of 1:1:2. Cell surface expression was assessed by Flow-cytometry with FITC-conjugated anti-HA antibody. Apoptosis assays were performed on HEK293 cells transfected with IRES-GFP constructs containing Kir6.1-WT or mutant (E332del or V346I) with SUR2A. After staining with PE-Annexin V and 7-AAD, the apoptotic cells were measured by Flow-cytometry within gated GFP (+) cells. Caspase-3/7 activity was measured with Apo-ONE Homogenous caspase 3/7 assay kit. The cell-counting studies showed that the cell surface expression of Kir6.1-WT was suppressed 40% to 70% when co-expressed with Kir6.1-E332del or Kir6.1-V346I. The apoptosis assay data indicated that the apoptotic ratio was increased significantly for E332del (38.9%) compared to WT (4.02%) and for V346I (11.2%) compared to WT (3.04%). The Caspase-3/7 activity was also increased 2.1 fold for E332del and 1.6 fold for V346I over WT.Conclusions: The loss-of-functional Kir6.1 KATP channel mutations found in SIDS display a dominant-negative effect on Kir6.1-WT channels and induce apoptosis in heterologous expression system.