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Rotator cuff injuries are a common clinical condition of the shoulder joint. Surgery that involves reattaching the torn tendon to its humeral head bony attachment has a somewhat lower success rate. The scar tissue formed during healing of the rotator cuff leads to poor tendon-related mechanical properties. To promote healing, a range of genetic interventions, as well as cell transplantation, and many other techniques have been explored. In recent years, the therapeutic promise of mesenchymal stem cells (MSCs) has been well documented in animal and clinical studies. Some data have suggested that MSCs can promote angiogenesis, reduce inflammation and cell proliferation and increase collagen deposition. These functions are likely paracrine effects of MSCs, particularly mediated through exosomes. Here, we review the use of MSCs-related exosomes in tissues and organs. We also discuss their potential utility for treating rotator cuff injuries, and explore the underlying mechanisms of their effects.

The use of epigenetic bromodomain inhibitors as anticancer therapeutics has transitioned from targeting bromodomain extraterminal domain (BET) proteins into targeting non-BET bromodomains. The two most relevant non-BET bromodomain oncology targets are cyclic AMP response element-binding protein (CBP) and E1A binding protein P300 (EP300). To explore the growing CBP/EP300 interest, we developed a highly efficient two-step synthetic route for dimethylisoxazole-attached imidazo[1,2-a]pyridine scaffold-containing inhibitors. Our efficient two-step reactions enabled high-throughput synthesis of compounds designed by molecular modeling, which together with structure-activity relationship (SAR) studies facilitated an overarching understanding of selective targeting of CBP/EP300 over non-BET bromodomains. This led to the identification of a new potent and selective CBP/EP300 bromodomain inhibitor, UMB298 (compound 23, CBP IC50 72 nM and bromodomain 4, BRD4 IC50 5193 nM). The SAR we established is in good agreement with literature-reported CBP inhibitors, such as CBP30, and demonstrates the advantage of utilizing our two-step approach for inhibitor development of other bromodomains.

Mitochondrial dysfunction contributes to osteoarthritis (OA) onset and progress. Mitochondrial dynamics, coupled with mitophagy, is critical for the maintenance of mitochondrial fitness, involving many cellular processes, such as proliferation and apoptosis. Excessive mechanical stress induces chondrocyte apoptosis; however, the effects of mechanical stress on mitochondrial dynamics remain elusive. In this study, we performed fluorescence staining, flow cytometry, transmission electron microscope, Western blot analysis, and RNA-sequencing to assess the effects of different strength of mechanical stimulation on mitochondrial functions of chondrocyte treated with interleukin-1β (IL-1β). We found that moderate mechanical stress reduced the IL-1β-induced apoptosis by maintaining mitochondrial function and scavenging the reactive oxygen species, while excessive mechanical stress induced strong mitochondrial dysfunction and apoptosis. Moreover, RNAsequencing revealed that mitophagy and mitochondrial dynamics were involved in the regulation of mechanical stress on chondrocyte biology. In addition to the elevated mitophagy, moderate mechanical stress also promoted mitochondrial dynamics by enhancing the expression of MFN1/2 and OPA1 and the translocation of dynamin-related protein 1 from the cytoplasm to the mitochondria. However, an uncoupling of mitochondrial dynamics, characterized by strongly elevated fission, resulted in the unfavorable apoptosis of excessive mechanical stress-stimulated chondrocytes. This study revealed the effects of mechanical stress upon mitochondrial dynamics in chondrocyte.

To evaluate the recovery of early urinary continence in patients with prostate cancer using a suprapubic catheter during Retzius-sparing robotic-assistant laparoscopic prostatectomy. From January 2018 to January 2019, 223 patients diagnosed with prostate cancer who underwent Retzius-sparing robotic-assistant laparoscopic prostatectomy in Diakonie Klinikum Stuttgart were involved in our study. From January 2018 to June 2018, patients (112 cases) only had an indwelling urinary catheter during Retzius-sparing robotic-assistant laparoscopic prostatectomy, while from July 2018 to January 2019, patients (111 cases) were offered an extra suprapubic catheter during operation. The recovery of early urinary continence of patients was mainly investigated one month later. The overall early urinary continence rate was 81.61%. Patients with suprapubic catheter had better urinary control results, compared to patients with only indwelling urinary catheter (87.39% vs 75.89%, p = 0.027). In addition, International Prostate Symptom Score and irritative subscore in patients with good urinary control were significantly lower than that in patients with urinary incontinence. Suprapubic catheter insertion (OR 0.395; 95% CI 0.190–0.821) and advanced pathological tumor stage (T3a–T4) (OR 2.061; 95% CI 1.008–4.217) were two independent influencing factors for early urinary continence recovery in patients who underwent Retzius-sparing robotic-assistant laparoscopic prostatectomy through multivariate logistic regression analysis. Suprapubic catheter insertion may be helpful for early urinary continence recovery in patients with Retzius-sparing Robotic-assistant laparoscopic prostatectomy. Advanced pathological tumor stage (T3a–T4) before Retzius-sparing robotic-assistant laparoscopic prostatectomy might be associated with poor urinary control.

Fungi are important resources for drug development, as they have a diversity of genes, that can produce novel secondary metabolites with effective bioactivities. Here, five depsidone-based analogs were isolated from the rice media of Chaetomium brasiliense SD-596. Their structures were elucidated using NMR and mass spectrometry analysis. Five compounds, including three new depsidone analogs, mollicellin S (1), mollicellin T (2), and mollicellin U (3), and two known compounds, mollicellin D (4) and mollicellin H (5), exhibited significant inhibition against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA), with MIC values ranging from 6.25 to 12.5 μg ml-1. Herein, we identified the predicted plausible biosynthetic cluster of the compounds and discussed the structure-activity relationship. Finally, we found that the introduction of aldehyde and methoxyl groups provide marked improvement for the inhibition against MRSA.

Background: Hepatitis B virus infection is associated with liver disease, including cancers. In this study, we assessed the power of sex-determining region Y (SRY)-related high-mobility group (HMG)-box 4(SOX4) gene to predict the clinical course of hepatocellular carcinoma (HCC). Methods: To evaluate the differential expression of SOX4 and its diagnostic and prognostic potential in HCC, we analyzed the GSE14520 dataset. Stratified analysis and joint-effect analysis were done using SOX4 and clinical factor. We then designed a nomogram for predicting the clinical course of HCC. Differential SOX4 expression and its correlation with tumor stage as well as its diagnostic and prognostic value were analyzed on the oncomine and GEPIA websites. Gene set enrichment analysis was explored as well as candidate gene ontology and metabolic pathways modulated by in SOX4 HCC. Results: Our analysis revealed that the level of SOX4 was significantly upregulated in tumor issue (P

The Human Leukocyte Antigen (HLA) genes, playing key roles in mediating the immune response, especially HLA class II alleles were suggested to play a role in the activation of autoreactive T-cells in aplastic anemia (AA). Previous studies in different ethnic groups have indicated that some of HLA-A,-B,-DRB1 alleles had a protective or susceptive association with the prevalence, pathogenesis and development of AA. HLA class II genes, especially HLA-DQB1 and -DPB1 alleles or haplotypes at high-resolution level associated with AA have not been fully identified in northern Chinese Han populations. The aim of this study was to identify association of the variations in HLA class II region with AA in northern Chinese Han population. A recent case-control study, including 96 AA patients and 824 healthy controls was performed. The high-resolution HLA genotyping was conducted by PCR-SBT, -SSO and NGS-ION S5TM platform. Based on genotypic data of the three loci, haplotype estimation was carried out. HLA-DRB1*15:01 (Pc = 2.87 × 10-3; OR = 2.11, 95% CI = 1.45–3.07) and HLA-DQB1*06:02 (Pc = 1.86 × 10-2; OR = 2.01, 95% CI = 1.32–3.06) were the risk and predisposition alleles to AA in northern Chinese Han after considering multiple testing. Moreover, the HLA-DRB1*15:01-DQB1*06:02 (Pc = 4.90 × 10-3; OR = 2.09, 95% CI = 1.37–3.19) and HLA-DRB1*14:05-DQB1*05:03 (Pc = 2.65 × 10-2; OR = 2.82, 95%CI = 1.45–5.50) haplotypes had direct strong relevance to AA and were the susceptible haplotypes. HLA-DPB1 alleles and 23 polymorphic amino acid residues spanning exon 2 ~ 4 of DPβ1 molecules have showed no statistically significant associations between AA and controls. The present findings establish a novel link between inherited HLA-DRB1,-DQB1,-DPB1 risk alleles and haplotypes in northern Chinese Han with AA, and open new avenues for development of targeted therapies to prevent or redirect immunopathology in AA.

This present study was designed to evaluate the immunomodulatory activity of Konjac glucomannan (KGM) on immunosuppressed mice induced by cyclophosphamide (CTX) treatment. The mice immunodeficiency model was established by CTX. KGM was used to modulate the activities of immunosuppressive mice. It was proved that KGM could promote the proliferation of lymphocyte, thymus, and spleen indices, and alleviate the atrophy of immune organs and weight loss. Besides, in mice serum, the levels of cytokines including TNF‐α, IgG, IL‐2, and the contents of hemolysin were also increased after treatment with KGM. Furtherly, in nonspecific immunity, KGM could enhance natural killer (NK) cell lethality and pinocytic activity of mouse peritoneal macrophages. Therefore, all of these results revealed that KGM could improve the reduced immunity of CTX‐induced mice via modulation innate immunity and adaptive immunity., This present study was designed to evaluate the immunomodulatory activity of Konjac glucomannan (KGM) on immunosuppressed mice induced by cyclophosphamide (CTX) treatment. It was found that KGM could improve the reduced immunocompetence of CTX‐induced mice via modulation innate immunity and adaptive immunity.

The transient receptor potential vanilloid type 4, TRPV4, is a polymodal cation channel which can be activated by diverse stimuli including mechanical, thermal and chemical cues. In the urinary bladder, TRPV4 is not only abundantly expressed in the urothelium but may also be localized in subepithelium, detrusor smooth muscles and afferent neurons. Emerging evidence indicates that the TRPV4 channel plays a sensory role in the uroepithelium, where it may regulate the release of sensory mediators such as ATP, which in turn modulates afferent nerve activity in response to bladder filling during the urination cycle. TRPV4 may also directly regulate detrusor contractility and the urothelial barrier function. Altered TRPV4 expression has been detected in various pathological bladder conditions. As such, TRPV4 may be a promising therapeutic target for bladder dysfunctions.

Despite advancements in treatment options, the overall cure and survival rates for non–small cell lung cancers (NSCLC) remain low. While small-molecule inhibitors of epigenetic regulators have recently emerged as promising cancer therapeutics, their application in patients with NSCLC is limited. To exploit epigenetic regulators as novel therapeutic targets in NSCLC, we performed pooled epigenome-wide CRISPR knockout screens in vitro and in vivo and identified the histone chaperone nucleophosmin 1 (Npm1) as a potential therapeutic target. Genetic ablation of Npm1 significantly attenuated tumor progression in vitro and in vivo. Furthermore, KRAS-mutant cancer cells were more addicted to NPM1 expression. Genetic ablation of Npm1 rewired the balance of metabolism in cancer cells from predominant aerobic glycolysis to oxidative phosphorylation and reduced the population of tumor-propagating cells. Overall, our results support NPM1 as a therapeutic vulnerability in NSCLC. Significance: Epigenome-wide CRISPR knockout screens identify NPM1 as a novel metabolic vulnerability and demonstrate that targeting NPM1 is a new therapeutic opportunity for patients with NSCLC.

Background This study was mainly to explore and study the potential application of lipoxygenases (ALOX) family genes in the diagnostic and prognostic values of colon adenocarcinoma (COAD). Methods Data sets related to the ALOX genes of COAD were obtained from The Cancer Genome Atlas and the University of California, Santa Cruz Xena browser. Then, the relevant biological information was downloaded from the public data platform. Finally, the bioinformatics technologies and clinical verification were employed to comprehensively analyze the potential values of ALOX genes. Results The Pearson correlation analysis indicated that there were correlations among ALOXE3, ALOX5, ALOX12, and ALOX12B. The diagnostic receiver operating characteristic (ROC) curves suggested that ALOXE3 and ALOX12 had significant diagnosis in COAD: ALOXE3; P

OBJECTIVES To validate the operational and diagnostic performances of a new device for transient elastography (TE), FibroTouch, for liver fibrosis in patients with chronic hepatitis B (CHB). METHODS In this prospective multicenter study, adult patients with CHB and valid liver pathological results were recruited to validate the operational and diagnostic performance of a TE device by FibroTouch for staging liver fibrosis. RESULTS In total, 517 patients with histologically proven CHB were enrolled. All had achieved at least 10 successful liver stiffness measurements (LSM), resulting in a success rate of 99.1% and reliable evaluations of 95.2%. Altogether 412 patients were included to analyze the diagnostic performance of FibroTouch. The area under the receiver operating characteristic curve for the LSM was 0.846 (95% confidence interval [CI] 0.808-0.880) for fibrosis stage ≥ F1, 0.850 (95% CI 0.811-0.883) for ≥ F2, 0.908 (95% CI 0.876-0.934) for ≥ F3 and 0.874 (95% CI 0.836-0.903) for F4. The diagnostic accuracy of LSM was superior to that of gamma-glutamyl transpeptidase-to-platelet ratio (GPR), aminotransferase-to-platelet ratio index (APRI), or fibrosis index based on 4 factors (FIB-4) index in staging fibrosis F2-F4 (P = 0.007 to

Immune-based rather than cancer-based classification systems are becoming an important additional component for prognostic prediction. Herein, we investigate the status of tumor-immune interaction and prognostic value of immune activity scores in bladder urothelial carcinoma (BLCA), kidney renal clear cell carcinoma (KIRC), and prostate adenocarcinoma (PRAD) from The Cancer Genome Atlas dataset and tracking the tumor immunophenotype platform. Traditional clinicopathological parameters still are effective predictors for prognosis. Comparison of 23-set, seven-step immune activity scores, and its distribution between favorable and adverse outcome groups within each cancer type and among cancer types show that each cancer has a distinct tumor-immune pattern. Adjusted overall immune activity scores based on the binary logistic regression analysis show a great discrimination ability in the progression-free survival (p = .0056 in BLCA, p < .0001 in KIRC, and p < .0001 in PRAD). K-mean cluster method and principal component analysis were performed to explore the cancer subtype. The results reveal that certain immune activity scores pattern such as high nature killer (NK) cell, T cell, macrophage, T helper type 1 (Th1) cell, and dendritic cell recruiting scores in BLCA, low NK cell, CD8 T cell, macrophage, T cell, Th1 cell recruiting scores in KIRC have a favorable clinical outcome comparing to other subtypes. Also, It is remarkable that a relatively small subtype of patients has a poor clinical prognosis in PRAD and this subtype features high CD4 T cell, macrophage, T cell, and regulatory T cell recruiting scores.

Background This meta-analysis aims to systematically evaluate the evidence for mindfulness-based stress reduction (MBSR) in cancer related fatigue (CRF). Material and methods In October 2018, PubMed, Embase, Cochrane Library, Clinical Trials, China Biomedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI) and China Science Periodical Database (CSPD) were searched for randomized controlled trials on MBSR in CRF patients. Literature screening and data extraction were conducted by two reviewers. Methodological quality evaluation was assessed by the Cochrane risk of bias tool. Revman 5.3.0 performs data analysis. The trial sequential analysis software estimated the required information size for each outcome indicator. Results There have been 5 studies included in this research for meta-analysis, 356 cases in the experimental group and 344 cases in the control group. The meta-analysis result indicates that: MBSR can reduce the cancer-related fatigue score of cancer patients, SMD = −0.51,95%CI [−0.81–0.20], P = 0.001, and the difference is statistically significant. The trial sequential analysis indicates that: The RIS required for the indicator to reach the level of significance test should be 1768. The sample size (700 cases) included in the study has not reached the RIS, but it has crossed the traditional threshold and the TSA threshold, indicating that the results tend to be stable. The grading results are shown as low-quality evidence. Conclusions This research has used evidence-based medicine to evaluate whether MBSR can alleviate CRF in cancer patients and provide evidence for the comprehensive intervention program for patients with cancer-related fatigue.

CRISPR-based genetic screening has revolutionized cancer drug target discovery, yet reliable, multiplex gene editing to reveal synergies between gene targets remains a major challenge. Here, we present a simple and robust CRISPR-Cas12a-based approach for combinatorial genetic screening in cancer cells. By engineering the CRISPR-AsCas12a system with key modifications to the Cas protein and its CRISPR RNA (crRNA), we can achieve high efficiency combinatorial genetic screening. We demonstrate the performance of our optimized AsCas12a (opAsCas12a) through double knockout screening against epigenetic regulators. This screen reveals synthetic sick interactions between Brd9&Jmjd6, Kat6a&Jmjd6, and Brpf1&Jmjd6 in leukemia cells., Reliable, multiplexed gene editing to uncover synergies between targets remains challenging. Here, the authors engineer AsCas12a and the crRNA to improve double knockout for synthetic sick/lethal interaction genetic screening.

Background and Aim In China, clinical experience with direct‐acting antiviral treatments for hepatitis C virus (HCV) infection is still emerging. C‐CORAL is a phase 3, multinational, placebo‐controlled, double‐blind trial of elbasvir/grazoprevir (EBR/GZR) in participants with HCV infection from the Asia‐Pacific region and Russia. Here, we report the data from participants enrolled in China. Methods Treatment‐naive participants with chronic HCV genotype (GT) 1, GT4, or GT6 infection were randomly assigned to receive 50 mg EBR/100 mg GZR for 12 weeks (immediate‐treatment group, ITG) or placebo followed by deferred treatment with EBR/GZR (deferred‐treatment group, DTG). The primary efficacy end‐point was sustained virologic response at 12 weeks after completing treatment (SVR12), and the primary safety end‐point was a comparison of safety between participants receiving EBR/GZR and placebo (NCT02251990; Protocol PN‐5172‐067). Results A total of 152 participants in China were randomly assigned (ITG, n = 115; DTG, n = 37). SVR12 was achieved in 96.7% (146/151) participants overall and in 97.3% (142/146) of those with GT1b infection. Four participants relapsed (GT1b, n = 3; GT6a, n = 1). Drug‐related AEs were reported in 25 (21.7%) and 9 (24.3%) participants receiving EBR/GZR and placebo, respectively; no drug‐related serious adverse events (AEs) occurred. Two (1.7%) participants receiving EBR/GZR had late hepatic transaminase elevations. Patient‐reported outcomes indicate improved quality of life at follow‐up week 4 in participants receiving EBR/GZR compared to placebo. Conclusion EBR/GZR administered for 12 weeks represents a highly effective and safe treatment option for Chinese individuals with HCV GT1 infection., A sustained virologic response was achieved by 96.7% (146/151) of treatment‐naive participants with hepatitis c virus infection receiving elbasvir/grazoprevir (50 mg/100 mg) for 12 weeks. Patient‐reported outcomes indicate improved quality of life following clearance of HCV.

Transcription is epigenetically regulated by the orchestrated function of chromatin-binding proteins that tightly control the expression of master transcription factors, effectors, and supportive housekeeping genes required for establishing and propagating the normal and malignant cell state. Rapid advances in chemical biology and functional genomics have facilitated exploration of targeting epigenetic proteins, yielding effective strategies to target transcription while reducing toxicities to untransformed cells. Here, we review recent developments in conventional active site and allosteric inhibitors, peptidomimetics, and novel proteolysis-targeted chimera (PROTAC) technology that have deepened our understanding of transcriptional processes and led to promising preclinical compounds for therapeutic translation, particularly in cancer.

Background To investigate the safety and efficacy of scleral concave pool trabeculectomy (SCPT) combined phacoemulsification for eyes with coexisting cataract and primary open-angle glaucoma (POAG). Methods This was a retrospective, controlled, interventional pilot case series. Thirty patients (30 eyes) were diagnosed with coexisting cataract and POAG between May 2015 and April 2018. Fourteen eyes underwent SCPT combined phacoemulsification were set as the study group, and 16 eyes received conventional phacotrabeculectomy were set as the control group. All patients were followed up for at least 6 months. The preoperative to postoperative changes in IOP, glaucoma medication requirements, BCVA, blebs functions, and adverse events were recorded. Results The groups were matched for baseline age, BCVA, IOP and types of IOP-lowering medications (all P > 0.05). At 6-month visit, there were no significant difference between control and study group in the improvement of BCVA (0.22 ± 0.24 versus 0.18 ± 0.26, P = 0.718), reduction of IOP (− 11.21 ± 8.61 mmHg versus − 9.19 ± 9.18 mmHg, P = 0.540) and the number of eyes that needed IOP-lowering medications (2 versus 3, P = 0.743). At the last visit, the rate of forming functioning blebs was significantly different between the study and control groups, (92.9% versus 68.7% respectively, P = 0.007). In the study group, 5 eyes developed hypotony, and 1 eye showed limited choroidal detachment, whereas in the control group 1 eye developed malignant glaucoma. All adverse events were successfully managed. Conclusion The SCPT combined phacoemulsification seems to be a safe and effective alternative to conventional phacotrabeculectomy for patients with POAG and visually significant cataract in the short-term.

Background Although circulating tumor cells (CTCs) are considered as a surrogate marker in monitoring disease progression and treatment response in late stage prostate cancer (PCa), its clinical impact in localized PCa remains unclear, indicating the limitation that is simply based on cell count. This perspective observational study aimed to detect the epithelial-to-mesenchymal transition (EMT) subtypes of CTCs in localized PCa and analyze their clinical relevance and application in predicting PCa stages before surgery compared with the Partin table. Patients and methods Between August 2017 and April 2019, 80 newly diagnosed localized PCa patients were enrolled in the study. Peripheral blood samples (5 mL) were collected prior to surgery. The CanPatrolTM CTC enrichment technique, a size-based isolation method, was used to detect the EMT CTCs. Clinical relevance of the CTCs was analyzed with Spearman's rank correlation test. Models to predict pathological were built with multivariate logistic regression. Receiver operating characteristic (ROC) curve and area under the curve (AUC) analysis were performed to evaluate the accuracy of the prediction model. Results CTCs were detected in 55% of all patients. The biophenotypic CTCs were most valuable and closely correlated with PSA, Gleason score, D'Amico risk classification, and pathological stage in localized PCa. The mesenchymal subtype was rare in this population but associated with seminal vesicle invasion, while the epithelial subtype had limited clinical significance. In addition, the biophenotypic CTCs combined with traditional clinical variables were analyzed by multivariate logistic regression to predict organ-confined disease before surgery, of which the AUC reached 0.818 and was superior to the Partin table 2017 in our cohort. Conclusion This study highlights the clinical impact of the biophenotypic CTCs in localized PCa, which was most closely related to clinical variables and could help to predict pathology outcomes before surgery.

We aimed to verify the expression status and diagnostic significance of isocitrate dehydrogenase 1 (IDH1) in non‐small‐cell lung cancer (NSCLC), especially during early stages. Serum IDH1 levels were measured by ELISA. A total of 1223 participants (660 patients with NSCLC, 276 healthy controls [HCs], 95 patients with benign pulmonary conditions [BPCs], 135 patients with other cancers [OCs], and 57 samples with interfering factors) were divided into a training cohort and a validation cohort according to 3 testing centers. The IDH1 concentrations in the NSCLC group were obviously higher than those in the control groups (P, This is the first large‐scale study aiming to verify the level and diagnostic value of serum isocitrate dehydrogenase 1 (IDH1) in non‐small‐cell lung cancer (NSCLC) with well‐designed controls in multicenter‐based cohorts. IDH1 showed high diagnostic efficiency in early stage NSCLC and high specificity for NSCLC comparing to other cancers.

AIM Genetic factors are believed to be implicated in the pathogenesis of relapsing polychondritis (RP). However, the molecular genetic determinants remain to be elucidated. This study aimed to detect the susceptibility genes of RP with whole-exome sequencing (WES) in a Chinese family and deepen our understanding of the pathogenesis of RP thereafter. METHOD A 32-year-old Chinese female proband with RP and her family including her mother with RP were enrolled in the study. The genomic DNA of 6 human subjects was extracted from peripheral blood and then gene allele mutations were identified using WES. Candidate variants with low frequency (

Background: It has been demonstrated that miRNAs play critical roles in the tumorigenesis and progression of various tumors.Objective: The purpose of this research was to determine the serum miR-63...

Effective harvesting procedure for adipose tissue is demanded by the affordable Good Manufacturing Practice-Compliant Production of clinical-grade adipose tissue-derived stem cells (hADSCs). Enzymatic digestion using collagenase is the most reliable method of adipose tissue-derived stem cells (hADSCs) isolation, while the optimized loading volume ratios of digestion to container during the shaking process of adipose tissue and collagenase mixture are still lacking. This study was conducted to determine the optimized loading volume ratio (mixture to container) for enzymatic digestion of Stromal/Stem Cells from lipoaspirate. Lipoaspirates were obtained from twelve women immediately after liposuction. Then tissue from each patient was divided into four groups according to different loading volume ratios in 50 ml centrifugal tube: 0.2 group, 0.4 group, 0.6 group, 0.8 group. Stromal vascular fractions (SVF) were obtained from each group, then total cell counts, viability and viable cell count were performed. hADSCs were harvested at passage (P) 2, whose morphologies, immunophenotypes, proliferation, and tri-differentiation abilities were compared. 0.4 loading volume ratio provided the highest cell yield, favorable viability and viable cell yield. The proliferation and triple differentiation ability of hADSCs obtained by 0.4 group was not inferior to that of other groups. Therefore, 0.4 may be the optimal loading volume ratio for hADSCs isolation from lipoaspirate by enzymatic digestion in current setting.

Flexor tendon injuries and tendinopathy are very common but remain challenging in clinical treatment. Exosomes-based cell-free therapy appears to be a promising strategy for tendon healing, while limited studies have evaluated its impacts on tenocyte biology. The objective of this study was to characterize a novel purified exosome product (PEP) derived from plasma, as well as to explore its cellular effects on canine tenocyte biology. The transmission electron microscope revealed that exosomes of PEP present cup-shaped structures with the diameters ranged from 80 to 141 nm, and the NanoSight report presented that their size mainly concentrated around 100 nm. The enzyme-linked immunosorbent assay kits analysis showed that PEP was positive for CD63 and AChE expression, and the cellular uptake of exosomes internalized into tenocyte cytoplasm was observed. The cell growth assays displayed that tenocyte proliferation ability was enhanced by PEP solution in a dose-dependent manner. Tenogenic phenotype was preserved as is evident by that tendon-related genes expression (SCX, COL1A, COL3A1, TNMD, DCN, and MKX) were expressed insistently in a high level, while tenocytes were treated with 5% PEP solution. Furthermore, migration capability was maintained and total collagen deposition was increased. More interesting, dexamethasone-induced cellular apoptosis was attenuated during the incubation of tenocytes with a 5% PEP solution. These findings will provide the basic understandings about the PEP, and support the potential use of this biological strategy for tendon healing.

To investigate the dry eye symptoms after cataract surgery in MGD patients and their relationships METHODS: The study included 115 patients (115 eyes) with age-related cataract that underwent uncomplicated cataract surgery, and the patients were divided into two groups according to the MGD diagnostic criteria: group A (MGD group) and group B (control group). Schirmer I test (ST-I), tear breakup time (TBUT), and corneal fluorescein staining (CFS) were performed preoperatively and at 3 days, 7 days, 14 days, and 30 days postoperatively. We also measured eyelid meibomian gland morphology, meibomian gland expression, and meibum character scores before and after the cataract surgery.Postoperatively, in group A, TBUT decreased and CFS scores increased significantly. ST-I increased in the early postoperative course but decreased later. The eyelid margin morphology scores and meibomian gland expression scores of group A significantly increased after the cataract operation. Thus, patients with MGD may have a greater chance of developing the dry eye disease after cataract surgery. Cataract surgery may aggravate the signs of MGD, and the severity of MGD may positively correlate with TBUT, CFS, and corneal lesions after surgery.The characteristics of dry eye after cataract surgery in patients with MGD are different from common cataract patients, changes in the early postoperative phase to the ocular surface were caused by surgical factors, and the damages to epithelial function in the later postoperative phase were mainly associated with the inflammation of the meibomian gland and eyelid.

Increasing attention has recently been focused on complex symbiotic associations, for instance coral and its symbionts. Sea cucumber, harboring diverse fungi, has also attracted more and more attention for their functional diversity. Here, secondary metabolites produced by Chaetomium globosum associated with sea cucumber, Apostichopus japonicus, were investigated using gene mining with third-generation sequencing technology (PacBio SMRT). Nine compounds, including one new compound cytoglobosin X (1), were isolated from cultures of Chaetomium globosum. Compound 1 was identified based on NMR data, HRESIMS, and ECD, and the absolute configurations were identified as 3S, 4R, 7S, 8R, 9R, 16S, 19S, 20S, and 23S. In an antimicrobial assay, compound 4 showed moderate activity against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus with MICs of 47.3 and 94.6 μM, respectively. Our results suggest that the microbiomes associated with sea cucumber could be an important resource for biodiversity and structural novelty, and the bioactive compounds may protect the host from pathogen microbial.

This study aimed to evaluate the taste profile of traditional Chinese Fuliji Red-cooked Chicken (FRCC) in five processing steps, consisting of raw, frying, stewing, vacuum-packaging and sterilization by measuring sodium chloride (NaCl), sugar, free amino acids (FAAs) and nucleotides, and by subjecting all the samples to an electronic tongue. Results showed that NaCl, 5ʹ-Inosine monophosphate (5ʹ-IMP), lysine (Lys), methionine (Met) and cysteine (Cys), which had relatively high contents, might be main taste compounds in FRCC. The NaCl content increased gradually from the frying to the sterilization step (P

The structure-properties relationships of native and debranched starch (DBS) were investigated by analyzing the results of DSC, XRD, NMR, HPAEC, FT-IR, SEM, hydrolysis, and digestibility properties. After debranching of starch in waxy rice, japonica rice, and indica rice, the linear short-chain molecules formed were easier to alignment and aggregation, and associate into a double helix. The crystalline structure of the rice starch after the pullulanase treatment was transformed from the type A to the type V crystalline by XRD measurement. Based on FT-IR and 13C NMR observations, molecular rearrangement and degree of order in starch granules increased. The DSC curve showed an increase in gelatinization temperature of debranched starch compared to native starch. Meanwhile, Solubility, water holding capacity and resistant starch content of DBS also raised. The study of structure- properties relationship provides a theoretical foundation for the development of foods or drugs with targeted functional properties. Abbreviations: WS: waxy rice starch; JS: Japonica rice starch; IS: Indica rice starch; DBWS: debranched waxy rice starch; DBJS: debranched Japonica rice starch; DBIS: debranched Indica rice starch; RDS: rapidly digestible starch; SDS: slowly digestible starch; RS: resistant starch; DP: degree of polymerization; DSC: differential scanning calorimertry; Gelatinization temperature at onset (To), peak (Tp), and end (Tc); ∆H: transition enthalpy; HPSEC: high-performance size-exclusion chromatography; ATR-FTIR: Attenuated total reflectance Fourier transform infrared spectroscopy; NMR: nuclear magnetic resonance; XRD: X-ray diffraction; SEM: Scanning electron microscopy; WHC: Water holding capacity.

Objective: The goal of our current study is to assess the immunohistochemical of p53, p21, nm23, and VEGF expression in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) prognosis after hepatectomy, as well as the prospective molecular mechanisms of prognostic indicator. Methods: There were 419 HBV-related HCC patients who were from southern China of Guangxi province and were used to evaluate the immunohistochemical expression for these biomarkers in prognosis. A genome-wide expression microarray dataset of HBV-related HCC were obtained from GSE14520. Results: In our study, the expression of p53, p21, and nm23 in cancer tissues of patients with hepatitis B-related hepatocellular carcinoma did not affected the clinical outcome of 2 years, 5 years or overall. Patients with high expression of VEGF had a worse overall survival after 2 years of surgery than patients with low expression (adjusted P=0.040, adjusted HR = 1.652, 95% CI = 1.024-2.665). Survival analysis of VEGF in GSE14520 cohort also demonstrated that VEGF mRNA expression also significantly associated with HBV-related HCC OS (adjusted P=0.035, adjusted HR =1.651, 95% CI =1.035-2.634). The prospective molecular mechanisms by co-expression analysis suggested that VEGF might be correlated to regulation of cell proliferation, cell growth and apoptotic process, Rap1 signaling pathway, HIF-1 signaling pathway, PPAR signaling pathway, cell cycle. Whereas the GSEA suggested that VEGF might involve in the regulation of HIF and HIF1A pathway, and TP53 regulation pathway. Conclusion: Our findings suggested that VEGF might be a prognostic indicator of HBV-related HCC, and we also identified the VEGF prospective molecular mechanisms through the whole genome co-expression and GSEA approaches.

Amplification of MYCN is a poor prognostic feature in neuroblastoma (NBL) indicating aggressive disease. We and others have shown BET bromodomain inhibitors (BETi) target MYCN indirectly by downregulating its transcription. Here we sought to identify agents that synergize with BETi and to identify biomarkers of resistance. We previously performed a viability screen of ∼1,900 oncology-focused compounds combined with BET bromodomain inhibitors against MYCN-amplified NBL cell lines. Reanalysis of our screening results prominently identified inhibitors of aurora kinase A (AURKAi) to be highly synergistic with BETi. We confirmed the anti-proliferative effects of several BETi+AURKAi combinations in MYCN-amplified NBL cell lines. Compared to single agents, these combinations cooperated to decrease levels of N-myc. We treated both TP53-wild type and mutant, MYCN-amplified cell lines with the BETi JQ1 and the AURKAi Alisertib. The combination had improved efficacy in the TP53-WT context, notably driving apoptosis in both genetic backgrounds. JQ1+Alisertib combination treatment of a MYCN-amplified, TP53-null or TP53-restored genetically engineered mouse model of NBL prolonged survival better than either single agent. This was most profound with TP53 restored, with marked tumor shrinkage and apoptosis induction in response to combination JQ1+Alisertib. BETi+AURKAi in MYCN-amplified NBL, particularly in the context of functional TP53, provided anti-tumor benefits in preclinical models. This combination should be studied more closely in a pediatric clinical trial.

Two papers, published in Science and Nature, report inhibitiors selectively targeting distinct bromodomains of BET proteins. These advanced chemical tools facilitate functional understanding of epigenetic regulators in normal and disease states and enable development of new medicine potentially with lower toxicity.

We recently reported that inhibition of BET bromodomain suppresses adipogenesis in vitro. In the present study we aimed to address whether BET bromodomain inhibition can suppress adipogenesis in vivo. Brd4fl/fl mice were crossed with B6.Cg-Tg(Fabp4-cre)1Rev/J mice to generate Brd4fl/+/Fabp4-cre mice. We used high fat diet (HFD, 45% fat) mice treated with vehicle (DMSO) or JQ1 (intraperitoneal, IP injection, 50 mg/kg/day), respectively, for 6 weeks. Body weight was measured once a week. Dual-energy X-ray absorptiometry was determined and brown adipose tissue was harvested at the end of the experiment. Partial deletion of Brd4 leads to the lower body weight. JQ1 treatment further confirmed that BET bromodomain inhibition suppresses body weight gain and decreases white adipose depots compared with the control mice. In addition, JQ1 treatment reduces the size of brown adipose tissue and impairs its thermogenesis. BET bromodomain inhibition suppresses adipogenesis in the mice.

Endoscopic transaxillary augmentation mammaplasty breast augmentation offers several advantages over other augmentation methods. Nonetheless, this procedure is fraught with some problems, including greater surgical trauma due to the longer separation area. We hypothesized that cold separation of the interpectoral space could reduce surgical injury in comparison to the electrosurgical method. This study aimed to compare the outcomes of endoscopic-assisted transaxillary augmentation mammaplasty using cold separation versus electrosurgical separation of the interpectoral space.In this prospective clinical trial, cold and electrosurgical separation of the interpectoral space were achieved using a separation shovel and monopolar electrotome, respectively. A total of 20 patients who visited our department in Beijing, China, for primary breast augmentation surgeries from October 1, 2017, and May 31, 2018, were included. The primary outcome was total postoperative drainage volume. The secondary outcomes were operative time, daily drainage volume, daily pain as assessed using the visual analogue scale (VAS), and reoperation rate. Quantitative data were compared using independent-samplesThe total drainage volume was significantly lower in the cold separation group than in the electrosurgical separation group (170.45 ± 75.40 mL vs 281.05 ± 148.43 mL;Cold separation is more conducive to reducing drainage, relieving postoperative pain, and causing less damage than the electrosurgical method in endoscopic-assisted transaxillary dual-plane augmentation mammaplasty.L’augmentation mammaire transaxillaire par voie endoscopique comporte plusieurs avantages par rapport aux autres méthodes d’augmentation. Cette intervention se heurte toutefois à certains problèmes, y compris des traumatismes chirurgicaux plus importants à cause de la zone de séparation plus longue. Les auteurs ont postulé que la séparation de l’espace interpectoral par le froid réduirait davantage la lésion que la méthode électrochirurgicale. La présente étude visait à comparer les résultats cliniques de l’augmentation mammaire transaxillaire assistée par endoscopie au moyen de la séparation par le froid plutôt que par la séparation électrochirurgicale de l’espace interpectoral.Dans la présente étude clinique prospective, la séparation par le froid et la séparation électrochirurgicale de l’espace interpectoral ont été assurées par une pince de séparation et une électrode monopolaire, respectivement. Au total, 20 participants qui ont consulté le département des auteurs à Beijing, en Chine, afin de subir une augmentation mammaire primaire entre le 1Le volume de drainage total était considérablement plus faible dans le groupe de séparation par le froid que dans celui de séparation électrochirurgicale (170,45 ± 75,40 mL par rapport à 281,05 ± 148,43 mL; P = 0,005). Le score d’ÉAV le premier jour postopératoire était considérablement plus faible dans le groupe de séparation par le froid que dans celui de séparation électrochirurgicale (6,45 ± 1,93 par rapport à 7,55 ± 1,43; P = 0,048). Deux réopérations (20 %) causées par la douleur postopératoire ou la rigidité de l’implant ont été exécutées dans le groupe de séparation électrochirurgicale.La séparation par le froid favorise la diminution du drainage, le soulagement de la douleur postopératoire et la réduction des dommages davantage que la méthode életrochirurgicale en cas d’augmentation mammaire transaxillaire biplan assistée par endoscopie.

Background Calcifying fibrous tumor (CFT) is a rare benign mesenchymal tumor that often occurs in deep soft tissue of children and young adults. CFT rarely occurs in the mediastinum. Case summary In this paper, we describe a 31-year-old male patient with CFT in the mediastinum. The patient did not have any symptoms, and the posterior mediastinal lesion was unintentionally found during routine re-examination of thyroid cancer. The tumor had no adhesion to the surrounding tissue and was successfully and completely removed. Pathology showed a large amount of collagen-rich fibrous connective tissue. There was scattered dystrophic calcification and gravel in the fibrous tissue and a small amount of lymphocyte and plasma cell infiltration and lymphoid follicle formation in the interstitial fluid. In addition, findings showed 20 IgG4+ plasma cells per high-powered field of the diseased tissue, an IgG4+/IgG ratio of about 20%, and normal serum IgG4 levels. The final diagnosis was CFT of the mediastinum (CFTM). No evidence of tumor recurrence was observed by computed tomography at 3 mo after surgery. Conclusion IgG4+ plasma cell enlargement may occur in CFTM, but clinical manifestations and serological tests suggest that it is not IgG4-related disease. We speculate that it may be an independent tumor subtype.

Identity determining transcription factors (TFs), or core regulatory (CR) TFs, are governed by cell-type specific super enhancers (SEs). Drugs to selectively inhibit CR circuitry are of high interest for cancer treatment. In alveolar rhabdomyosarcoma, PAX3-FOXO1 activates SEs to induce the expression of other CR TFs, providing a model system for studying cancer cell addiction to CR transcription. Using chemical genetics, the systematic screening of chemical matter for a biological outcome, here we report on a screen for epigenetic chemical probes able to distinguish between SE-driven transcription and constitutive transcription. We find that chemical probes along the acetylation-axis, and not the methylation-axis, selectively disrupt CR transcription. Additionally, we find that histone deacetylases (HDACs) are essential for CR TF transcription. We further dissect the contribution of HDAC isoforms using selective inhibitors, including the newly developed selective HDAC3 inhibitor LW3. We show HDAC1/2/3 are the co-essential isoforms that when co-inhibited halt CR transcription, making CR TF sites hyper-accessible and disrupting chromatin looping., Core regulatory transcription factors are usually regulated by cell-type specific super enhancers (SEs). Here, the authors screen for chemical probes able to distinguish between SE-driven and promoter-driven transcription and find that histone deacetylases are selectively required for core regulatory transcription.

BET-bromodomain inhibition (BETi) has shown pre-clinical promise for MYC-amplified medulloblastoma. However, the mechanisms for its action, and ultimately for resistance, have not been fully defined. Here, using a combination of expression profiling, genome-scale CRISPR/Cas9-mediated loss of function and ORF/cDNA driven rescue screens, and cell-based models of spontaneous resistance, we identify bHLH/homeobox transcription factors and cell-cycle regulators as key genes mediating BETi’s response and resistance. Cells that acquire drug tolerance exhibit a more neuronally differentiated cell-state and expression of lineage-specific bHLH/homeobox transcription factors. However, they do not terminally differentiate, maintain expression of CCND2, and continue to cycle through S-phase. Moreover, CDK4/CDK6 inhibition delays acquisition of resistance. Therefore, our data provide insights about the mechanisms underlying BETi effects and the appearance of resistance and support the therapeutic use of combined cell-cycle inhibitors with BETi in MYC-amplified medulloblastoma., BET-bromodomain inhibitors could be used to treat medulloblastoma tumors with Myc amplifications. Here, the authors show that both the response and resistance to BET inhibitors in mice is mediated by bHLH/homeobox transcription factors.

Proteins usually associate with other molecules physically to execute their functions. Identifying these interactions is important for the functional analysis of proteins. Previously, we reported the parallel analysis of translated ORFs (PLATO) to couple ribosome display of full-length ORFs with affinity enrichment of mRNA/protein/ribosome complexes for the “bait” molecules, followed by the deep sequencing analysis of mRNA. However, the sample processing, from extraction of precipitated mRNA to generation of DNA libraries, includes numerous steps, which is tedious and may cause the loss of materials. Barcoded PLATO (PLATO-BC), an improved platform was further developed to test its application for protein interaction discovery. In this report, we tested the antisera-antigen interaction using serum samples from patients with inclusion body myositis (IBM). Tripartite motif containing 21 (TRIM21) was identified as a potentially new IBM autoantigen. We also expanded the application of PLATO-BC to identify protein interactions for JQ1, single ubiquitin peptide, and NS5 protein of Zika virus. From PLATO-BC analyses, we identified new protein interactions for these “bait” molecules. We demonstrate that Ewing sarcoma breakpoint region 1 (EWSR1) binds to JQ1 and their interactions may interrupt the EWSR1 association with acetylated histone H4. RIO kinase 3 (RIOK3), a newly identified ubiquitin-binding protein, is preferentially associated with K63-ubiquitin chain. We also find that Zika NS5 protein interacts with two previously unreported host proteins, par-3 family cell polarity regulator (PARD3) and chromosome 19 open reading frame 53 (C19orf53), whose attenuated expression benefits the replication of Zika virus. These results further demonstrate that PLATO-BC is capable of identifying novel protein interactions for various types of “bait” molecules. Keywords: Barcoded PLATO, Protein interaction, Ubiquitin-binding protein, Bromodomain inhibitor JQ1, Zika virus

BackgroundThe concept of mixed adenoneuroendocrine carcinoma (MANEC) was introduced in the 2010 World Health Organization classification of digestive neuroendocrine neoplasms. Bile duct invasion by MANEC is exceptionally rare. We herein report a case of MANEC with invasion of multiple bile ducts. Case presentation: A 60-year-old man presented with a 7-day history of upper abdominal pain, and a mass in the cystic duct was suspected based on computed tomography findings. The patient underwent resection of the extrahepatic bile ducts with concomitant radical lymphadenectomy and Roux-en-Y cholangiojejunostomy. Large cell neuroendocrine carcinoma was detected in a component of the resected tumor. According to the pathological and immunohistochemical features of the tumor, the final histopathological diagnosis was a biliary MANEC, tumor stage T2N0M1 (Stage IIIC). The patient recovered uneventfully and was discharged from the hospital 10 days after surgery.ConclusionsWe have described a rare case of extrahepatic MANEC invading multiple bile ducts, with particular emphasis on the physician’s awareness of MANEC and its optimal treatment. MANEC arising from extrahepatic bile ducts is rare, and surgical resection is the most effective treatment method.

OBJECTIVE: Distress tolerance and anxiety sensitivity may differentiate among internalizing disorders, though few studies have examined differential associations of distress tolerance and anxiety sensitivity with depression and anxiety symptoms while adjusting for their intercorrelation. METHOD: In an adolescent genetic epidemiological sample (age 15 to 21), the present study (N = 848, 56.97% female) examined concurrent associations of distress tolerance and anxiety sensitivity with internalizing psychopathology (i.e., symptoms of depression, anxiety, and general stress) at baseline and prospective, predictive associations of baseline distress tolerance and anxiety sensitivity with internalizing psychopathology at 2-year follow-up. Additionally, the present study assessed distress tolerance with two laboratory-based tasks, a carbon dioxide challenge and the mirror-tracing task, to distinguish between tolerance of physiological and cognitive distress, respectively. RESULTS: Elevated anxiety sensitivity was broadly associated with elevated symptoms of internalizing psychopathology at baseline and prospectively predicted elevated depression, anxiety, and stress symptoms at 2-year follow-up. Higher tolerance of cognitive distress was associated with lower concurrent anxiety symptoms but not with anxiety symptoms at follow-up. CONCLUSIONS: The present results clarify previously mixed findings; during adolescence, anxiety sensitivity showed broad concurrent and prospective associations with internalizing disorder risk whereas distress tolerance, specifically regarding cognitive distress, was associated with only elevated concurrent anxiety symptoms.

The present study aimed to investigate the clinical significance and prospective molecular mechanism of cystatin (CST) genes in patients with hepatitis B virus (HBV)‑related hepatocellular carcinoma (HCC). The role of CST genes in the molecular mechanism of HCC was revealed through bioinformatics analysis. The clinical significance of CST genes was investigated using GSE14520‑derived data from patients with HBV‑related HCC. Gene set enrichment analysis (GSEA) was used to identify pathways in which the CST genes were enriched, as well as the association between these pathways and HCC. The expression levels of CST1, CST2, CST5, CSTA and CSTB genes were higher in HCC tissue compared with in normal tissue; conversely, CST3 and CST7 were reduced in HCC tissue. Subsequent receiver operating characteristic analysis of the CST genes demonstrated that CST7 and CSTB genes may function as potential diagnostic markers for HCC. Furthermore, the expression levels of CST6 and CST7 were strongly associated with recurrence‑free survival and overall survival of patients with HBV‑related HCC. GSEA of the CST genes revealed that CST7 was significantly enriched in tumor evasion and tolerogenicity, cancer progenitors, liver cancer late recurrence, liver cancer progression and several liver cancer subclasses. In addition, CST genes demonstrated homology in terms of protein structure and were revealed to be strongly co‑expressed. The present findings suggested that CST7 and CSTB genes may serve as potential prognostic and diagnostic biomarkers for HCC.

Verticillium wilt caused by the soil-borne fungus Verticillium dahliae is a devastating disease of cotton, leading to serious loss of lint yield in China. It is important to characterize the prevailing populations of V. dahliae for understanding disease development. In this study, the representative 74 V. dahliae isolates from 28 different cotton regions in three major cotton-production areas in China were identified for their pathotypes, races, vegetative compatibility groups (VCGs) and genetic diversity by specific PCR, amplified fragment length polymorphism (AFLP) and inter simple sequence repeat (ISSR) analysis. Of 74 isolates, 60 isolates (81%) were defoliating (D) pathotype strains, especially in Yellow River/Yantze River Valley region. Seventy isolates belonged to race 2, whereas no race 1 specific fragment was amplified. These isolates were grouped into five VCGs: VCG1A (48), VCG2A/VCG4B (3), VCG2B824 (6), VCG6 (5). The D pathotype and VCG1A isolates have become the dominant strains, moderately and highly virulent isolates were widely distributed in three major cotton-growing areas in China. High level of polymorphic markers was obtained by AFLP and ISSR analyses. Through comparison of the obtained dendrograms by unweighted paired group method with arithmetic averages cluster analysis (UPGMA), the V. dahliae isolates were divided into two major groups and five subgroups. The obvious correlations with the geographic origin of the isolates between AFLP and ISSR were revealed, with significant diversity among V. dahliae isolates. In addition, AFLP dendrogram showed relationships with pathotypes. The results offered a better understanding of the epidemiology of Verticillium wilt in China.

Flexor tendon injury is often associated with suboptimal outcomes and results in substantial digit dysfunction. Stem cells have been isolated from several experimental animals for the growing interest and needs of utilizing cell-based therapies. Recently, turkey has been developed as a new large animal model for flexor tendon research. In the present study, we reported the isolation and characterization of bone marrow-derived mesenchymal stem cells (BMSCs) from 8- to 12-month-old heritage-breed turkeys. The isolated cells demonstrated fibroblast-like morphology, clonogenic capacity, and high proliferation rate. These cells were positive for surface antigens CD90, CD105, and CD44, but were negative for CD45. The multipotency of turkey BMSCs was determined by differentiating cells into osteogenic, adipogenic, chondrogenic, and tenogenic lineages. There was upregulated gene expression of tenogenic markers, including mohawk, tenomodulin, and EGR1 as well as increased collagen synthesis in BMP12 induced cells. The successful isolation and verification of bone marrow-derived MSCs from turkey would provide opportunities of studying cell-based therapies and developing new treatments for tendon injuries using this novel preclinical large animal model. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1419-1428, 2019.

Rationale: Hepatitis B virus (HBV) is a leading cause of liver diseases. HBV covalently closed circular DNA (cccDNA) is a critical obstacle of complete elimination by anti-HBV therapy. HBV cccDNA accumulates in nucleus as a chromatin-like cccDNA minichromosome assembled by histones and non-histones. However, the underlying mechanism of modulation of cccDNA minichromosome in hepatocytes is poorly understood. Methods: A human liver-chimeric mouse model was established. The cccDNA-ChIP, Southern blot analysis, confocal assays, RIP assays and RNA pull-down assays, et al. were performed to assess the mechanism of assembly and epigenetic regulation of cccDNA minichromosome in human liver-chimeric mouse model, human primary hepatocytes (PHH), dHepaRG, HepG2-NTCP cell lines and clinical liver tissues. Results: Importantly, the expression levels of HAT1, CAF-1 and lncRNA HULC were significantly elevated in the liver from HBV-infected human liver-chimeric mice. Strikingly, the depletion of HAT1 reduced HBV replication and cccDNA accumulation, and impaired the assembly of histone H3/H4 and the deposition of HBx and p300 onto cccDNA to form cccDNA minichromosome in the cells. Mechanically, chromatin assembly factor-1 (CAF-1) was involved in the events. Interestingly, HAT1 modified the acetylation of histone H3K27/H4K5/H4K12 on cccDNA minichromosome. Moreover, lncRNA HULC-scaffold HAT1/HULC/HBc complex was responsible for the modification on cccDNA minichromosome. Additionally, HBV activated HAT1 through HBx-co-activated transcriptional factor Sp1 in a positive feedback manner. Conclusion: HAT1 signaling contributes to assembly and epigenetic regulation of HBV cccDNA minichromosome.

PGGFRα+ preadipocytes are the major subpopulations that can regenerate into adipocytes. Two different types of macrophages exist in the fat tissue: the classically activated macrophage (M1) and the alternatively activated macrophage (M2). In this study, we investigated whether M1/M2 macrophages play distinct roles in adipogenic differentiation of PDGFRα+ preadipocytes. Mouse preadipocytes and macrophages were isolated from C57BL/6 male mice of 6–8 weeks. The culture supernate of M1 and M2 macrophages was collected and co-cultured with the PDGFRα+ preadipocytes. After 3 days, Oil Red O staining was used to evaluate to adipogenic differentiation of PDGFRα+ preadipocytes and the expression of adipogenic-related transcription factors (C/EBP-α, PPARγ) were also tested. The results showed that when cultured in vitro, M1 macrophages could significantly suppress the adipogenesis of PDGFRα+ preadipocytes as well as the C/EBP-α and PPARγ expression, but M2 macrophages did not have significant influence on the adipogenesis of PDGFRα+ preadipocytes nor on C/EBP-α and PPARγ expression compared with the control group. M1 macrophages significantly suppress PDGFRα+ preadipocyte adipogenesis which provides a possible way to improve adipogenesis and fat retention after fat-free grafting by mitigating acute inflammation and manipulating M1 macrophage levels. 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 .

Background Transforming growth factor-β (TGF-β) pathway presents dysregulation in pathological scarring and mediates hypertrophic scar (HS) formation. Objectives The study aims to analyze the potential mechanism of long non-coding RNA NORAD (LncRNA NORAD) and microRNA (miR-26a) regulation of the TGF-β pathway in hypertrophic scar fibroblasts (HSFs). Material and methods Hypertrophic scar tissues were collected and assayed for LncRNA NORAD, miR-26a, transforming growth factor β receptor I (TGF-βR1) and TGF-βR2, with enzyme-linked immunosorbent assay (ELISA) or qualitative polymerase chain reaction (qPCR). LncRNA NORAD interfering plasmids were transfected into HSFs and induced with TGF-β1. Cell Counting Kit-8 (CCK-8) assays were performed to assess HSF proliferation, and flow cytometry to analyze apoptosis and the cell cycle. TGF-βR1, TGF-βR2, Smad2, and p-Smad2 levels were detected using western blot (WB). The related proteins (p21, cyclin D1 and cyclin-dependent kinase 4 (CDK4)) regulating the cell cycle, and apoptosis-related proteins (caspase-3 and Bcl-2) were also detected using WB. The binding sites of miRNA-26a and LncRNA NORAD, TGF-βR2, or UBE3A were predicted using Starbase and confirmed with dual luciferase reporter assay. RNA immunoprecipitation (RIP) was utilized to explore the interplay of miR-26a with its target genes. Results LncRNA NORAD is decreased, miR-26a is increased and TGF-β receptors show abnormal expression in scar tissue. LncRNA NORAD knockdown inhibits proliferation of HSF cells induced by TGF-β1 treatment. In addition, cell apoptotic levels are markedly increased and cell numbers in G0/G1 phase are increased. Moreover, the TGF-β/Smad pathway is regulated by decreasing endogenous LncRNA NORAD levels, possibly by affecting the relative levels of TGF-βR1. p21 is notably upregulated, while cyclin D1 and CDK4 are downregulated. Apoptosis-related proteins are significantly affected. LncRNA NORAD may act as a sponge, binding miR-26a and changing its expression. Finally, RIP shows that miR-26a targets the 3'UTRs of TGF-βR2 and UBE3A. Conclusions LncRNA NORAD regulates HSF proliferation via miR-26a mediating the regulation of TGF-βR2/R1. LncRNA NORAD/miR-26a could be a potential target for treating HS.

Background: Chronic prostatitis is hard to be identified in benign prostatic hyperplasia (BPH) patients in clinical works. This study aimed to diagnose chronic prostatitis in BPH patients by noninvasive methods.Methods: The research was carried out on the BPH patients who received transurethral resection of prostate at Xinhua hospital from January 2014 to Jul 2015. Before operation, patients were asked for medical history and required to receive physical examination, serum sample collection including PSA, sex hormones, inflammatory cytokines, metabolic panel and transrectal ultrasonography. According to histological results, the patients were divided into 2 group of BPH with/without prostatitis. Logistic regression was used to find the risk factors of chronic prostatitis.Results: As a result, 181 men with an average age of 72.15±8.41 years were enrolled in this study, including 116 patients with prostatitis and 65 patients without prostatitis. The storage sub-score (9.24±1.55 vs 8.52±1.63, p=0.009), PSA (8.23±7.69 vs 4.92±3.84, p=0.005) and IL-2R (531.96±200.75 vs 434.11±153.54, p=0.001) were significantly higher in patients with prostatitis than those without prostatitis. Based on logistic regression analysis, the above three parameters were also the risk factors of BPH with prostatitis. The diagnostic model was calculated as: 0.317* storage sub-score+0.092* PSA+0.003* IL-2R-4.296. The AUC was 0.725. Conclusions: Histological prostatitis in BPH patients can be diagnosed by the combination of serum IL-2R, PSA and storage sub-score. Identification of chronic prostatitis among patients with BPH is beneficial for medical decisions, which can more efficiently alleviate urinary symptoms and reduce the risk of disease progression.

Objectives Our study aimed to investigate the correlation of prostatic morphological parameters and benign prostatic hyperplasia (BPH) clinical progression in aging Chinese men. Methods In this retrospective study, a total of 1038 patients were reviewed. Prostatic morphology was measured by transrectal ultrasound (TRUS). Detailed medical history of all candidates was recorded and analyzed after being classified by specific prostatic measurements. Univariate and multivariate logistic regression analyses were used to estimate the correlation between variables. Results The cumulative incidence of BPH clinical progression was 63.68% (661/1038) in the study population. Prostate volume (PV), transitional zone volume (TZV), transitional zone index (TZI), and intravesical prostatic protrusion (IPP) were all positively associated with BPH progression (all p 60 ml, TZV > 15 ml, TZI > 0.5, or IPP > 5 mm had a significantly higher possibility of overall BPH clinical progression (adjusted odds ratio (OR): 2.485, 1.678, 1.886, and 1.924, respectively; 95% confidence interval (CI): 1.559-3.960, 1.131-2.489, 1.379-2.579, and 1.357-2.728, correspondingly). Conclusion Prostatic morphological parameters are significantly associated with BPH clinical progression. Patients with larger prostatic morphological parameters are more easily prone to clinical progress. As a result, reasonable managements should be timely considered for those patients before clinical progression occurs.

Neurogenesis in the developing neocortex begins with the generation of the preplate, which consists of early-born neurons including Cajal-Retzius (CR) cells and subplate neurons. Here, utilizing the Ebf2-EGFP transgenic mouse in which EGFP initially labels the preplate neurons then persists in CR cells, we reveal the dynamic transcriptome profiles of early neurogenesis and CR cell differentiation. Genome-wide RNA-seq and ChIP-seq analyses at multiple early neurogenic stages have revealed the temporal gene expression dynamics of early neurogenesis and distinct histone modification patterns in early differentiating neurons. We have identified a new set of coding genes and lncRNAs involved in early neuronal differentiation and validated with functional assays in vitro and in vivo. In addition, at E15.5 when Ebf2-EGFP+ cells are mostly CR neurons, single-cell sequencing analysis of purified Ebf2-EGFP+ cells uncovers molecular heterogeneities in CR neurons, but without apparent clustering of cells with distinct regional origins. Along a pseudotemporal trajectory these cells are classified into three different developing states, revealing genetic cascades from early generic neuronal differentiation to late fate specification during the establishment of CR neuron identity and function. Our findings shed light on the molecular mechanisms governing the early differentiation steps during cortical development, especially CR neuron differentiation., Author summary Neural stem cells and progenitor cells in the embryonic brain give rise to neurons following a precise temporal order after initial expansion. Early-born neurons including Cajal-Retzius (CR) cells and subplate neurons form the preplate in the developing cerebral cortex, then CR neurons occupy the layer 1, playing an important role in cortical histogenesis. The molecular mechanisms governing the early neuronal differentiation processes remain to be explored. Here, by genome-wide approaches including bulk RNA-seq, single-cell RNA-seq and ChIP-seq, we comprehensively characterized the temporal dynamic gene expression profile and epigenetic status at different stages during early cortical development and uncovered molecularly heterogeneous subpopulations within the CR cells. We revealed CR neuron signatures and cell type-specific histone modification patterns along early neuron specification. Using in vitro and in vivo assays, we identified novel lncRNAs as potential functional regulators in preplate differentiation and CR neuron identity establishment. Our study provides a comprehensive analysis of the genetic and epigenetic programs during neuronal differentiation and would help bring new insights into the early cortical neurogenesis process, particularly the differentiation of CR neurons.

For controlling recent COVID-19 outbreaks around the world, many countries have implemented suppression and mitigation interventions. This work aims to conduct a feasibility study for accessing the effect of multiple interventions to control the COVID-19 breakouts in the UK and other European countries, accounting for balance of healthcare demand. The model is to infer the impact of mitigation, suppression and multiple rolling interventions for controlling COVID-19 outbreaks in the UK, with two features considered: direct link between exposed and recovered population, and practical healthcare demand by separation of infections. We combined the calibrated model with COVID-19 data in London and non-London regions in the UK during February and April 2020. Our finding suggests that rolling intervention is an optimal strategy to effectively control COVID-19 outbreaks in the UK for balancing healthcare demand and morality ratio. It is better to implement regional based interventions with varied intensities and maintenance periods. We suggest an intervention strategy named as “Besieged and rolling interventions” to the UK that take a consistent suppression in London for 100 days and 3 weeks rolling intervention in other regions. This strategy would reduce the overall infections and deaths of COVID-19 outbreaks, and balance healthcare demand in the UK.

BACKGROUND: While acute urinary retention (AUR) is a severe complication of benign prostatic hyperplasia (BPH), it can also indicate the progression of this common disease in aging men. This study aimed at exploring the possible relationship between serum interleukin 6 (sIL-6) and AUR in BPH patients. METHODS: A cross-sectional study was conducted based on 256 elderly men with BPH in China. The association between the sIL-6 level and the occurrence of AUR was evaluated by univariate and multivariable logistic regressions. The receiver operating characteristic (ROC) curve was utilized to determine the discriminant validity of the sIL-6 level and the optimal cut-off value. RESULTS: The concentration of sIL-6 was significantly elevated in the AUR group (P