DSCR1-1 attenuates osteoarthritis-associated chondrocyte injury by regulating the CREB1/ALDH2/Wnt/ß-catenin axis: An in vitro and in vivo study.

The progression of osteoarthritis (OA) includes the initial inflammation, subsequent degradation of the extracellular matrix (ECM), and chondrocyte apoptosis. Down syndrome candidate region 1 (DSCR1) is a stress-responsive gene and expresses in varied types of cells, including chondrocytes. Bioinformatics analysis of GSE103416 and GSE104739 datasets showed higher DSCR1 expression in the inflamed cartilage tissues and chondrocytes of OA. DSCR1 had two major isoforms, isoform 1 (DSCR1-1) and isoform 4 (DSCR1-4). We found that DSCR1-1 had a faster (in vitro) and higher expression (in vivo) response to OA compared to DSCR1-4. IL-1ß-induced apoptosis, inflammation, and ECM degradation in chondrocytes were attenuated by DSCR1-1 overexpression. DSCR1-1 triggered the phosphorylation of cAMP response element-binding 1 (CREB1) at 133 serine sites by decreasing calcineurin activity. Moreover, activated CREB1 moved into the cell nucleus and combined in the promoter regions of aldehyde dehydrogenase 2 (ALDH2), thus enhancing its gene transcription. ALDH2 could recover Wnt/ß-catenin signaling transduction by enhancing phosphorylation of ß-catenin at 33/37 serine sites and inhibiting the migration of ß-catenin protein from the cellular matrix to the nucleus. In vivo, adenoviruses (1 × 108 PFU) overexpressing DSCR1-1 were injected into the articular cavity of C57BL/6 mice with medial meniscus surgery-induced OA, and it showed that DSCR1-1 overexpression ameliorated cartilage injury. Collectively, our study demonstrates that DSCR1-1 may be a potential therapeutic target of OA.

Effects of nanographene oxide on adipose-derived stem cell cryopreservation.

Cryoinjury mitigation is key in cell cryopreservation. Here, we aimed to assess the effectiveness of nanographene oxide (nano-GO) for improving cryoprotectant agents (CPAs) in human adipose stem cell (hADSC) cryopreservation. For in vitro experiments, nano-GO (5 µg/mL) was added to the CPAs in the control, and passage (P) 2 hADSCs were collected and cryopreserved for around two weeks. We compared cytotoxicity, cell viability, immunophenotypes, proliferation, cell apoptosis, and tri-lineage differentiation. In vivo, studies used lipoaspirate to create non-enriched or hADSC-enriched fat tissues by combining it with PBS or hADSCs cryopreserved with the aforementioned CPAs. Each nude mouse received a 0.3 mL subcutaneous injection of the graft. At 12 weeks, the grafts were harvested. Histology, adipocyte-associated genes and protein, vascular density and angiogenic cytokines, macrophage infiltration, and inflammatory cytokines were analyzed. Nano-GO CPA contributed to increased cell viability, improved cell recovery, and lowered levels of early apoptosis. Nano GO at concentrations of 0.01-100 µg/mL caused no cytotoxicity to hADSCs. The absence of nano GOs in the intracellular compartments of the cells was confirmed by transmission electron microscopy. The fat grafts from the CPA-GO group showed more viable adipocytes and significantly increased angiogenesis compared to the PBS and CPA-C groups. Adding hADSCs from the CPA-GO group to the graft reduced macrophage infiltration and MCP-1 expression. Nano-GO plays an anti-apoptotic role in the cryopreservation of hADSCs, which could improve the survival of transplanted fat tissues, possibly via improved angiogenesis and lower inflammatory response in the transplanted adipose tissue.

Impaired neuronal macroautophagy in the prelimbic cortex contributes to comorbid anxiety-like behaviors in rats with chronic neuropathic pain.

A large proportion of patients with chronic pain experience co-morbid anxiety. The medial prefrontal cortex (mPFC) is proposed to underlie this comorbidity, but the molecular and neuronal mechanisms are not fully understood. Here, we reported that impaired neuronal macroautophagy in the prelimbic cortical (PrL) subregion of the mPFC paralleled the occurrence of anxiety-like behaviors in rats with chronic spared nerve injury (SNI). Intriguingly, such macroautophagy impairment was mainly observed in a FOS/c-Fos+ neuronal subpopulation in the PrL. Chemogenetic inactivation of this comorbid anxiety-related neuronal ensemble relieved pain-induced anxiety-like behaviors. Rescuing macroautophagy impairment in this neuronal ensemble relieved chronic pain-associated anxiety and mechanical allodynia and restored synaptic homeostasis at the molecular level. By contrast, artificial disruption of macroautophagy induced early-onset co-morbid anxiety in neuropathic rats, but not general anxiety in normal rats. Taken together, our work identifies causal linkage between PrL neuronal macroautophagy dysfunction and comorbid anxiety in neuropathic pain and provides novel insights into the role of PrL by differentiating its contribution in pain-induced comorbid anxiety from its modulation over general anxiety-like behaviors.Abbreviation: AAV: adeno-associated viruses; ACC: anterior cingulate cortex; ATG5: autophagy related 5; ATG7: autophagy related 7; ATG12: autophagy related 12; CAMK2/CaMKII: calcium/calmodulin-dependent protein kinase II; CNO: clozapine-N-oxide; CQ: chloroquine; DIA: data independent acquisition; DIO: double floxed inverse orf; DLG4/PSD-95: discs large MAGUK scaffold protein 4; Dox: doxycycline; GABA: γ-aminobutyric acid; GFP: green fluorescent protein; GO: gene ontology; Gi: inhibitory guanine nucleotide-binding proteins; HsCHRM4/M4D: human cholinergic receptor muscarinic 4; HsSYN: human synapsin; KEGG: Kyoto encyclopedia of genes and genomes; LAMP1: lysosomal-associated membrane protein 1; LC3-II: PE conjugated microtubule-associated protein 1 light chain3; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; mPFC: medial prefrontal cortex; P2A: 2A self-cleaving peptide; PPI: protein-protein interaction networks; PrL: prelimbic cortex; RBFOX3/NeuN: RNA binding protein, fox-1 homolog (C. elegans) 3; rtTA: reverse tetracycline-transactivator; SDS-PAGE: sodium dodecylsulfate-polyacrylamide gel electrophoresis; SHANK3: SH3 and multiple ankyrin repeat domains 3; SLC1A1/EAAC1: solute carrier family 1 (neuronal/epithelial high affinity glutamate transporter, systemXag), member 1; SNAP23: synaptosomal-associated protein 23; SNI:spared nerve injury; SQSTM1/p62: sequestosome 1; SYT3: synaptotagmin 3; TRE: tetracycline-responsive element; TRE3G: third-generation tetracycline-responsive element.

Comparison of the quality of Nurse-Led palliative care with standard medical care during six months in 405 patients with Parkinson's disease and burdens of their Caregivers: A retrospective study at a single center in China.

BACKGROUND: Palliative care is mainly used to improve the quality of life of patients with chronic diseases by addressing their medical conditions and psychological problems. End-stage Parkinson's disease (PD) is also a progressive disease like cancer and could be managed by palliative care. This study was conducted at a single center in China and aimed to compare the quality of nurse-led palliative care with standard medical care during six months in 405 patients with Parkinson's disease (PPD) and their caregivers using the Chinese version of the 39-item Parkinson's Disease Questionnaire (PDQ-39) and the Chinese Zarit Burden Interview (ZBI) scale. METHODS: PPD (stage 2-5) received nurse-led palliative care (NP cohort, 103 patients; 103 caregivers) or neurologist-led standard care (NS cohort, 134 patients; 134 caregivers), or primary care practitioner-led usual care (PS cohort, 168 patients; 168 caregivers) for six months. RESULTS: Before the health professional-led care (BN), the PDQ-39 score of PPD was 68 (71-64) and their caregivers had 54.86 ± 7.64 a ZBI scale. After 6-months of the health professional-led care (AN), the PDQ-39 score of PPD and a ZBI scale of their caregivers decreased for the NP cohort as compared to those of BN condition and those of patients in the NS and PS cohorts at AN condition (p < 0.001 for all). CONCLUSIONS: The quality of life of PPD must be improved and the burden on their caregivers must be relieved. Nurse-led palliative care successfully improved the quality of life of PPD and reduced their caregiver burden.

Transcriptomic and machine learning analyses identify hub genes of metabolism and host immune response that are associated with the progression of breast capsular contracture.

Capsular contracture is a prevalent and severe complication that affects the postoperative outcomes of patients who receive silicone breast implants. At present, prosthesis replacement is the major treatment for capsular contracture after both breast augmentation procedures and breast reconstruction following breast cancer surgery. However, the mechanism(s) underlying breast capsular contracture remains unclear. This study aimed to identify the biological features of breast capsular contracture and reveal the potential underlying mechanism using RNA sequencing. Sample tissues from 12 female patients (15 breast capsules) were divided into low capsular contracture (LCC) and high capsular contracture (HCC) groups based on the Baker grades. Subsequently, 41 lipid metabolism-related genes were identified through enrichment analysis, and three of these genes were identified as candidate genes by SVM-RFE and LASSO algorithms. We then compared the proportions of the 22 types of immune cells between the LCC and HCC groups using a CIBERSORT analysis and explored the correlation between the candidate hub features and immune cells. Notably, PRKAR2B was positively correlated with the differentially clustered immune cells, which were M1 macrophages and follicular helper T cells (area under the ROC = 0.786). In addition, the expression of PRKAR2B at the mRNA or protein level was lower in the HCC group than in the LCC group. Potential molecular mechanisms were identified based on the expression levels in the high and low PRKAR2B groups. Our findings indicate that PRKAR2B is a novel diagnostic biomarker for breast capsular contracture and might also influence the grade and progression of capsular contracture.

Effectiveness of a New Enzyme-Free Method for the Preparation of a Decellularized Adipose-Derived Matrix.

BACKGROUND: Decellularized adipose-derived matrix (DAM) represents a new alternative to tissue fillers. The function of DAM is closely associated with the decellularization technique used for its preparation. However, most techniques are time-consuming and expensive, and this might reduce the popularity of DAM. OBJECTIVES: The study aimed to investigate an enzyme-free adipose decellularization method and generate a DAM capable of adipose tissue regeneration. METHODS: DAMs prepared by the enzyme-free and Flynn's methods were compared and co-cultured with human adipose-derived stem cells (hADSCs) to investigate cytocompatibility. Adipose tissue formation was evaluated by injecting the DAMs into the backs of nude mice over 4 weeks. Samples were harvested for gross and perilipin immunohistochemistry analysis at 1 and 4 weeks. RESULTS: The enzyme-free method is effective for adipose decellularization because it removes adipocytes and preserves the microstructure. In vitro, the DAM made by the enzyme-free method could support the attachment, growth, proliferation, and differentiation of hADSCs, and promote the enhanced secretion of vascular endothelial growth factor by hADSCs; this DAM also induced the formation and maturity of adipocytes in vivo. CONCLUSIONS: This study describes a highly effective enzyme-free method for adipose tissue decellularization that also promotes adipocyte formation and adipose tissue volume stability in vitro and in vivo, resulting in a new alternative tissue filler.

Effect of radiation sterilization on the ability to induce adipose regeneration in vivo in decellularized adipose-derived matrix.

BACKGROUND: Decellularized adipose-derived matrix (DAM), a biological scaffold that can induce adipose regeneration. The balance between its sterilization efficiency and its ability to maintain in situ adipose regeneration should be considered in terminal sterilization. The purpose of this study was to investigate the effects of radiation sterilization of cobalt-60 (60 Co)with different doses on adipogenesis induced by different forms of DAM, so as to reduce radiation dose under the premise of safe and effective sterilization and ensure adipogenesis induced by DAM in vivo. METHODS: High dose (25 kGy) and low dose (5 kGy) radiation were used to sterilize freeze-dried and wet DAM, respectively. The sterilization efficiency, macro and micro characteristics, mechanical and mechanical properties of DAM were compared, and then implanted into the immunocompromised mice to evaluate the adipose regeneration. RESULTS: Under the two radiation doses, no microbial growth was found in the freeze-dried and wet DAM sterility tests, and no significant changes were observed in the macro and micro structures. In terms of mechanical properties, the elastic modulus of high dose freeze-dried DAM decreased significantly (p < 0.001). In vivo animal experiments, the freeze-dried DAM irradiated with high dose almost completely lost its function of adipogenesis in vivo. Although the wet DAM irradiated with high dose could induce fat regeneration in the early stage, the adipocyte deformation and atrophy appeared in the later stage. The freeze-dried and wet DAM after low dose irradiation was similar to the wet DAM without irradiation in the blank control, which could maintain excellent adipogenic and angiogenic functions in vivo. CONCLUSION: High dose 60 Co irradiation can completely destroy the ability of freeze-dried DAM to induce adipose regeneration in situ, while low dose irradiation (5 kGy) can effectively sterilize the DAM without damaging in vivo induced adipose regeneration. Radiation has more damage to freeze-dried DAM than wet DAM in adipogenesis properties.

GuiLu-ErXian Glue extract promotes mesenchymal stem cells (MSC)-Induced chondrogenesis via exosomes release and delays aging in the MSC senescence process.

ETHNOPHARMACOLOGICAL RELEVANCE: The treatment of osteoarthritis (OA) patients is a challenging problem. Mesenchymal stem cells (MSCs) are multipotent cells and play key roles in regenerative medicine for cartilage degeneration. GuiLu-ErXian Glue (GLEXG) is an herbal remedy widely used in traditional Chinese medicine to treat joint pain and disability in elderly OA patients. However, the mechanisms of how GLEXG affects MSCs-induced chondrogensis remains to be elucidated. AIM OF THE STUDY: The aim of this study was to investigate the effects of GLEXG on MSC-derived chondrogenesis, both in vitro and in vivo and its potential mechanisms. METHODS: Using human MSC (hMSCs) as in vitro model, the effects of HPLC-profiled GLEXG water extract on chondrogenic differentiation were investigated by 3D spheroid cultures under chondrogenesis-inducing medium (CIM) condition. The chondrogenesis process was evaluated by measuring the sphere sizes, chondrogenesis-related genes expression by reverse transcription real-time PCR that targeted type II/X collagens, SOX9, aggrecan, and protein expression by immunostaining. Anti-TGF-ß1 neutralization antibody was used for mechanistic study. Mono-iodoacetate (MIA) induced OA joint was used to evaluate the effects of GLEXG on in vivo model. MSCs-derived exosomes were purified for proteomics study and senescence process was evaluated by cumulative population doublings and senescence-associated ß-Galactosidase staining. RESULTS: The results showed that GLEXG enhanced hMSCs chondrogenesis and upregulated RNA expression of type II/X collagen, SOX9 and aggrecan at 0.1 µg/mL, 0.3 µg/mL in vitro. In vivo, GLEXG at the dose of 0.3 µg intraarticular (i.a.) injection rescued the MIA-induced cartilage defect. Proteomics and ingenuity pathway analysis obtained from MSCs-released exosomes suggested that senescence pathway was less activated in GLEXG group than in vehicle group. Besides, GLEXG was able to increase cumulative population doubling and delayed hMSCs senescence process after four passages in cultures. CONCLUSION: we conclude that GLEXG promotes in vitro MSC-induced chondrogenesis possibly via exosomes release and delays aging in the MSC senescence process and that treatment with GLEXG (0.3 µg, i.a.) rescued cartilage defects in rat OA knee model.

Cervical microendoscopic laminoplasty-induced clinical resolution of disc herniation in patients with single- to three-level myelopathy.

This study aimed to explore the effects on resorption of cervical disc herniation (CDH) and clinical outcomes of surgery. Cervical microendoscopic laminoplasty (CMEL), which is commonly preferable to anterior corpectomy and fusion, was applied to patients with 1- to 3-level degenerative cervical myelopathy (DCM). DCM patients with 1-3 levels DCM underwent either conservation treatment or CMEL. In conservation-treated patients (53 cases), CDH volume remained unchanged with no improvement in JOA and VAS scores. Conversely, 63 patients with 1-3 levels DCM were prospectively enrolled and exhibited a profound decrease in CDH volume: 89.1% of CDHs (123/138) regressed over 10%, 64.5% of CDHs (89/138) regressed over 25%, while 27.5% and 6.5% of CDHs (38/138 and 9/138) largely regressed over 50% and 75%, respectively. Meanwhile, the JOA and VAS scores were improved in different ways. Intriguingly, CDH volume changes correlated significantly with elevations in JOA scores, indicating an association of clinical CDH resolution with neurological recovery. We showed that CMEL induced clinically related diminishment of CDH and alleviation of clinical symptoms in patients with 1- to 3-level myelopathy and that it could help avoid anterior dissection of the disc to some extent.

Generating Self-Shaped 2D Aluminum Oxide Nanopowders.

The thermal-assisted exfoliation phenomena of boehmite particles under moderate heating rates were examined. The exfoliation that generated flakes of 5−6 nm in thickness can be achieved because of the perfect cleavage on the boehmite particles that are stripped when thermal treatments bring about dehydration and γ-Al2O3 formation in sequential phase transformation of boehmite. Examinations of the exfoliation effects were carried out on calcined boehmite single crystal particles, which were about 500 nm in diameter, and obtained at three heating rates 0.5, 1.0, and 2.0 °C/min with the heating schedules. The TEM techniques, BET-N2 measurements, XRD-Scherrer equation, and AFM images were employed in the examination. That the BET values increased as increasing of exfoliated flakes reflected two stages of exfoliation. In the beginning stage, during which the BET values were <40 m2/g, the exfoliation resulted from the stress produced by dehydration. In the second stage, the increased rate of surface area was due to the additional force, which originated from the γ-Al2O3 formation. Exfoliation occurred on the cleavage planes {010}, the side pinacoid of the boehmite particle. The generation of flakes resulted in the thinning of boehmite particles. Some of the flakes preserved the external form of boehmite crystals. From the surface energy evaluations of boehmite and γ-Al2O3, it can be inferred that exfoliation is a natural way of thermal treatment.

A Two-Step Technique for Correction of Severe Inverted Nipples with Minimally Invasive Procedures.

BACKGROUND: Congenital inverted nipple is not uncommon in Asian females, which impairs the patient's psychology and breastfeeding function. Although various methods have been reported in references, no consensus has been reached on an ideal treatment for severe inverted nipples. OBJECTIVES: This study aimed to propose a minimally invasive and reliable method that could correct severe inverted nipples, reduce their recurrence rate, and create a definite nipple neck for a natural shape. METHODS: We designed a two-step technique to correct severe inverted nipples, including congenital grade III inversion and recurrent grade III inversion with or without scar adhesion after correction with different methods from other hospitals. A retrospective study was performed on 38 patients (68 nipples) who underwent the new two-step technique in our department. The follow-up time was at least 6 months. Patient demographics, operation details, and complication rate were documented. RESULTS: A total of 68 nipples in 38 patients (bilateral: 30 patients; unilateral: 8 patients) received correction, among which there were four recurrent cases from other hospitals treated with different methods. In this study, the total recurrence rate was 7.89%. In these cases, the nipple exhibited the appearance of Grade I inversion, but not back to the preoperative state of Grade III. There was one case that suffered exposure of thread knot. CONCLUSIONS: The two-step technique is a minimally invasive method to successfully correct severe inverted nipples with a low recurrence rate and achieve a natural shape with a definite nipple neck. LEVEL OF EVIDENCE IV: 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 .

Multi-functional gene ZNF281 identified as a molecular biomarker in soft tissue regeneration and pan-cancer progression.

Regeneration and tumorigenesis are indicated as related processes, while regeneration leads to life and the outcome of tumorigenesis is death. Here, we show the upregulation of zfp281 (zinc finger 281) in our adipose de novo regeneration model through RNA-seq analysis. Then, we validated the upregulation of zfp281 in adipose regeneration via immunofluorescence. Following that, we found that ZNF281 (the human homolog of Zfp281) was upregulated in most types of cancer and related to worse prognosis in 10 tumors. We further investigated the role of ZNF281 in cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), pancreatic adenocarcinoma (PAAD), and stomach adenocarcinoma (STAD) and confirmed the high accuracy in the clinical diagnostic feature. Beyond that, based on these three types of cancers, we analyzed the ZNF281-related tumor immune infiltration and DNA methylation sites and finally built risk prediction models for future disease diagnosis. Taken together, our findings provide new insights into the dual role of ZNF281, and we found that it was a potential biomarker for regeneration and tumor prognosis.

Active compound identification by screening 33 essential oil monomers against Botryosphaeria dothidea from postharvest kiwifruit and its potential action mode.

The antifungal activity of postharvest kiwifruit against the pathogen Botryosphaeria dothidea was evaluated for 33 essential oil monomers. The possible mechanism for the known active compounds were further assessed in this study. The results show all the EO components exhibit inhibitory effects on the pathogen to different degrees except for Farnesol. Carbon chain length and C2-C3 double bonds had a great effect on the antifungal activities of aldehydes. Of all of these, carvacrol had the strongest antifungal activity with EC50 of 12.58 µL/L and EC90 of 22.08 µL/L. Carvacrol also exhibits significant inhibitory effects on the pathogen, both in vivo and in vitro. Carvacrol evidently alters the hyphal morphology of B. dothidea and severely damages cell membrane and inhibits the formation of lipid components on the membrane. As cell membrane permeability increases, intracellular homeostasis including ion and biomacromolecules were destroyed by carvacrol. Furthermore, carvacrol appears to significantly inhibit mitochondrial activity and respiration rates, resulting in cell death of B. dothidea. Our results provide evidence that carvacrol could be a very useful compound for controlling postharvest rot soft in kiwifruit.

Evidence of Browning of White Adipocytes in Poorly Survived Fat Grafts in Patients.

BACKGROUND: Browning adipocytes induced by burn and cancer were assumed less viable and more prone to necrosis for their hypermetabolic properties. Recent studies have shown browning of white adipose after fat engraftment in mice. OBJECTIVES: The authors sought to evaluate whether fat transfer could induce browning biogenesis in fat grafts in humans and if it is associated with graft necrosis. METHODS: Necrotic adipose grafts were excised from 11 patients diagnosed with fat necrosis after fat grafting or flap transfer. Non-necrotic fat grafts were from 5 patients who underwent revisionary surgeries after flap transfer. Histology and electronic microscopy as well as protein and gene expression of browning-related marker analyses were performed. RESULTS: Fat grafts with necrosis demonstrated a higher gene expression level of uncoupling protein-1 (greater than fivefold increase, **P < 0.01), a master beige adipocyte marker, than non-necrotic fat grafts. Electronic microscopy and histology showed that browning adipocytes were presented in necrotic adipose in patients. CONCLUSIONS: Fat transfer induced browning adipocytes in patients and was evident in patients with postgrafting necrosis.

Browning of White Adipocytes in Fat Grafts Associated With Higher Level of Necrosis and Type 2 Macrophage Recruitment.

BACKGROUND: Induced browning adipocytes were assumed less viable and more prone to necrosis for their hypermetabolic property. A previous study showed that browning of adipocytes was more evident in fat grafts with necrosis in humans. OBJECTIVES: The authors aimed to estimate whether fat transfer-induced browning biogenesis was associated with necrosis and its potential inflammation mechanisms in murine models. METHODS: Human subcutaneous adipose from thigh or abdomen of 5 patients via liposuction was injected in 100 µL or 500 µL (n = 20 per group) into the dorsal flank of 6- to 8-week-old female nude mice fed with normal chow diet and harvested after 2, 4, 8, and 12 weeks. Control groups did not receive any grafting procedures (sham operation), where lipoaspirates were analyzed immediately after harvest. Histology and electronic microscopy, immunological analyses of browning markers, necrosis marker, and type I/II macrophages markers in mice were performed. RESULTS: Histology and electronic microscopy showed browning adipocytes in fat grafts with a higher level of necrosis (0.435 ±â€…0.017 pg/mL for cleaved caspase-3, **P < 0.01), IL-6 (749.0 ±â€…134.1 pg/mL,***P < 0.001) and infiltration of type 2 macrophage profiles in mice (twofold increase, *P < 0.05). CONCLUSIONS: Browning of adipocytes induced by fat transfer in mice is in parallel with post-grafting necrotic levels associated with elevated interleukin-6 and activated type 2 macrophage profiles, which promote browning development.

Synthesis and evaluation of new compounds bearing 3-(4-aminopiperidin-1-yl)methyl magnolol scaffold as anticancer agents for the treatment of non-small cell lung cancer via targeting autophagy.

Magnolol and honokiol are the two major active ingredients with similar structure and anticancer activity from traditional Chinese medicine Magnolia officinalis, and honokiol is now in a phase I clinical trial (CTR20170822) for advanced non-small cell lung cancer (NSCLC). In search of potent lead compounds with better activity, our previous study has demonstrated that magnolol derivative C2, 3-(4-aminopiperidin-1-yl)methyl magnolol, has better activity than honokiol. Here, based on the core of 3-(4-aminopiperidin-1-yl)methyl magnolol, we synthesized fifty-one magnolol derivatives. Among them, compound 30 exhibited the most potent antiproliferative activities on H460, HCC827, H1975 cell lines with the IC50 values of 0.63-0.93 µM, which were approximately 10- and 100-fold more potent than those of C2 and magnolol, respectively. Besides, oral administration of 30 and C2 on an H460 xenograft model also demonstrated that 30 has better activity than C2. Mechanism study revealed that 30 induced G0/G1 phase cell cycle arrest, apoptosis and autophagy in cancer cells. Moreover, blocking autophagy by the autophagic inhibitor enhanced the anticancer activity of 30in vitro and in vivo, suggesting autophagy played a cytoprotective role on 30-induced cancer cell death. Taken together, our study implied that compound 30 combined with autophagic inhibitor could be another choice for NSCLC treatment in further investigation.

Does Water-Jet Force Affect Cryopreserved Adipose-Derived Stem Cells? Evidence of Improved Cell Viability and Fat Graft Survival.

BACKGROUND: Adipose tissue harvested by liposuctions is an available source of adipose-derived stem cells (ASCs). Water-jet-assisted liposuction is a favorable method for fat collection with little mechanical damage. This study aimed to investigate whether or not the water-jet-assisted liposuction made a difference in the biological characteristics of cryopreserved ASCs and fat graft survival in cell-assisted lipotransfer. METHODS: Human lipoaspirates were obtained from the abdomen or thighs of 20 female participants for body contouring. A single surgeon randomly harvested 50 mL of adipose tissue by the water-jet-assisted liposuction and the conventional liposuction, respectively. Adipose-derived stem cells were isolated from lipoaspirates and then cryopreserved for 4 weeks. Cryopreserved ASCs were used to examine the surface markers, cell proliferation, migration, and adipogenic differentiation in vitro. The fat survival of ASCs-enriched grafts from different liposuctions was measured in animal models. RESULTS: The cryopreserved ASCs with the water-jet assistance had better capacities of cell proliferation, migration, and adipogenic differentiation and achieved a better survival result of ASCs-enriched fat grafting. CONCLUSIONS: Cryopreservation of ASCs with the water-jet force showed more excellent biological characteristics. The water-jet-assisted liposuction was superior to the conventional liposuction in obtaining ASCs and fat survival of coimplantation with grafts.

LncRNA TTN-AS1 promotes the progression of oral squamous cell carcinoma via miR-411-3p/NFAT5 axis.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a common kind of squamous cell carcinoma of the head and neck, which is a threat to public health. Long noncoding RNAs (lncRNAs) are associated with the development of various diseases, including cancers. LncRNA titin antisense RNA 1 (TTN-AS1) is known as a crucial regulatory factor in several cancers. Nevertheless, the specific functions of TTN-AS1 in OSCC remains obscure. METHODS: The expression of TTN-AS1 in OSCC samples or cells was analyzed through qRT-PCR. Colony formation assay, EdU assay, flow cytometry assay, TUNEL assay and wound healing assay were conducted to estimate the functions of TTN-AS1 in OSCC cells. RIP and luciferase reporter assays were utilized to detect the interaction between TTN-AS1 and miR-411-3p as well as between miR-411-3p and NFAT5. RESULTS: TTN-AS1 expression was stronger in OSCC cells. Knockdown of TTN-AS1 effectively restrained cell proliferation and migration but had inductive role in apoptosis. Moreover, TTN-AS1 could function as the miR-411-3p sponge in OSCC and miR-411-3p exerted the inhibitory functions on OSCC cell growth. In addition, NFAT5 was proven as the target of miR-411-3p. Rescue assay indicated that overexpressing NFAT5 could reverse the inhibitory function of TTN-AS1 depletion on cell growth. CONCLUSION: lncRNA TTN-AS1 contributed to the progression of OSCC via miR-411-3p/NFAT5 axis.

Effects of Harvest Sites on Cryopreserved Adipose-Derived Stem Cells and ASC-Enriched Fat Grafts.

BACKGROUND: Enrichment of adipose-derived stem cells (ASCs) with fat grafts has demonstrated benefit for graft retention and histologic appearance. There is no consensus on the optimal harvest site for adipose-derived stem cells. This study aimed to investigate the effects of harvest sites on the characteristics of cryopreserved adipose-derived stem cells and the graft retention of cell-assisted lipotransfer. METHODS: Lipoaspirates were harvested from 18 healthy volunteers who underwent liposuctions for body contouring. Twenty milliliters of lipoaspirates was, respectively, obtained from four sites, including the upper limb, abdomen, waist, and thighs, by the Coleman technique. Adipose-derived stem cells were ex vivo cultured and cryopreserved for four weeks. The biological characteristics of ASCs from four harvest sites were analyzed: MSC surface markers, cell proliferation, migration ability, and multipotential differentiation. The fat grafts were co-implanted with ASCs from four harvest sites and injected subcutaneously in mice. The ASC-enriched fat grafts were analyzed three months after transplantation. RESULTS: Cryopreserved ASCs from the abdomen and thighs maintained more significant cell proliferation, migration ability, and differentiation potential, compared with cells from the upper limb and waist. Moreover, we achieved better graft retention of cell-assisted fat grafts with cryopreserved ASC from the abdomen and thighs. CONCLUSIONS: The harvest site of adipose tissue affects the cellular activity and differentiation potential of cryopreserved ASCs. Improved understanding of harvest sites for ASCs can optimize the outcomes of cell-assisted fat grafts. Fat grafts enriched with cryopreserved ASCs from the abdomen or thighs are the optimal choices. LEVEL OF EVIDENCE V: 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 .