Titanium-doped phosphate glasses containing zinc and strontium applied in bone regeneration.

Phosphate bioactive glass has been studied for its advanced biodegradability and active ion release capability. Our previous research found that phosphate glass containing (P2O5)-(Na2O)-(TiO2)-(CaO)-(SrO) or (ZnO) showed good biocompatibility with MG63 and hMSCs. This study further investigated the application of 5 mol% zinc oxide or 17.5 mol% strontium oxide in titanium-doped phosphate glass for bone tissue engineering. Ti-Ca-Na-Phosphate glasses, incorporating 5% zinc oxide or 17.5% strontium oxide, were made with melting quenching technology. The pre-osteoblast cell line MC3T3-E1 was cultured for indirect contact tests with graded diluted phosphate glass extractions and for direct contact tests by seeding cells on glass disks. The cell viability and cytotoxicity were analysed in vitro over 7 days. In vivo studies utilized the tibial defect model with or without glass implants. The micro-CT analysis was performed after surgery and then at 2, 6, and 12 weeks. Extractions from both zinc and strontium phosphate glasses showed no negative impact on MC3T3-E1 cell viability. Notably, non-diluted Zn-Ti-Ca-Na-phosphate glass extracts significantly increased cell viability by 116.8% (P < 0.01). Furthermore, MC3T3-E1 cells cultured with phosphate glass disks exhibited no increase in LDH release compared with the control group. Micro-CT images revealed that, over 12 weeks, both zinc-doped and strontium-doped phosphate glasses demonstrated good bone incorporation and longevity compared to the no-implant control. Titanium-doped phosphate glasses containing 5 mol% zinc oxide, or 17.5 mol% strontium oxide have promising application potential for bone regeneration research.

Preoperative homocysteine modifies the association between postoperative C-reactive protein and postoperative delirium.

Background: Homocysteine and C-reactive protein (CRP) may serve as biomarkers of postoperative delirium. We set out to compare the role of blood concentration of homocysteine versus CRP in predicting postoperative delirium in patients. Materials and methods: In this prospective observational cohort study, the plasma concentration of preoperative homocysteine and postoperative CRP was measured. Delirium incidence and severity within 3 days postoperatively were determined using the Confusion Assessment Method and Confusion Assessment Method-Severity algorithm. Results: Of 143 participants [69% female, median (interquartile range, 25th-75th) age of 71 (67-76) years] who had knee or hip surgery under general anesthesia, 44 (31%) participants developed postoperative delirium. Postoperative plasma concentration of CRP was associated with postoperative delirium incidence [adjusted odds ratio (OR) per one standard deviation change in CRP: 1.51; 95% Confidence Interval (CI): 1.05, 2.16; P = 0.026], and severity [in which each one standard deviation increase in postoperative CRP was associated with a 0.47 point (95% CI: 0.18-0.76) increase in the severity of delirium, P = 0.002] after adjusting age, sex, preoperative Mini-Mental State Examination score and the days when postoperative CRP was measured. A statistically significant interaction (adjusted P = 0.044) was also observed, in which the association between postoperative plasma concentration of CRP and postoperative delirium incidence was stronger in the participants with lower preoperative plasma concentrations of homocysteine compared to those with higher preoperative levels. Conclusion: Pending validation studies, these data suggest that preoperative plasma concentration of homocysteine modifies the established association between postoperative plasma concentration of CRP and postoperative delirium incidence.

Treatment of postoperative delirium with continuous theta burst stimulation: study protocol for a randomised controlled trial.

INTRODUCTION: Postoperative delirium is one of the most common postoperative complications among elderly patients (65 years old or older). However, there are no effective treatments for this condition. Recent research suggests that continuous theta burst stimulation (cTBS), a non-invasive brain stimulation, can reduce pain level, improve cognitive function and affective symptoms in multiple diseases or dysfunctions, including anxiety disorders, major depressive disorder, sleep disorders and pain. But the potential benefits of cTBS in reducing postoperative delirium have not been investigated. Therefore, we propose determining whether cTBS can prevent and/or treat postoperative delirium in senior patients. METHODS AND ANALYSIS: The study will be a double-blind, randomised controlled trial. Participants (65 years old or older) undergoing scheduled orthopaedic surgery (≥2 hours, general anaesthesia) will be randomised to receive either cTBS or sham stimulation with a focal figure-of-eight coil over the right dorsolateral prefrontal cortex at 80% of the resting motor threshold. Every patient will receive 2-3 sets of stimulations during postoperative days (40 s per session, 3 sessions per set, 1 set per day). Participants will be assessed twice daily by a research assistant blinded to allocation. The primary outcome will be the incidence of postoperative delirium measured by the Confusion Assessment Method on postoperative days 1, 2 and 3. The secondary outcomes will be the severity and duration of postoperative delirium, cognitive function, pain, sleep quality, activities of daily living, length of hospital stay, discharge-to-facility or home, and rate of complication and mortality during the hospital stay. ETHICS AND DISSEMINATION: Ethical approval has been obtained from the ethics committee of Shanghai 10th People's Hospital. The principal investigator will submit a research progress report to the ethics committee regularly. All participants will provide written informed consent. Study results will be published in a peer-reviewed journal. TRIAL REGISTRATION NUMBER: NCT04661904.

SOX11 promotes osteoarthritis through induction of TNF-α.

OBJECTIVE: Osteoarthritis (OA) is a degenerative disease and the molecular mechanism of OA remains unclear. Transcription factor SOX11 has been proved to be involved in the development progress of OA. The present study aimed to evaluate the potential function of SOX11 during the development of OA. METHODS: SOX11 expression in patients with OA and health donator was determined with qRT-PCR. Subsequently, in vitro OA model was established by treating the chondrocyte cells CHON-001 with IL-1ß. Next, we validated the function of SOX11 in in vitro OA model by using siRNAs. Finally, the relationship between SOX11 and TNF-α was explored. RESULTS: SOX11 was upregulated in patients with OA and in IL-1ß treated cells. IL-1ß significantly increased both the mRNA and protein levels of MMP13 and cleaved caspase 3, while decreased collagen II and aggrecan in CHON-001 cells. In addition, knockdown of SOX11 could significantly decrease IL-1ß-induced apoptosis in CHON-001 cells. Meanwhile, IL-1ß induced OA like phenomenon was significantly reversed by siRNA interference. Moreover, inhibition of SOX11 decreased the level of TNF-α in patients with OA and in IL-1ß treated cell supernatant. CONCLUSION: Inhibition of SOX11 could improve IL-1ß-induced OA like phenomenon in CHON-001 cells, which suggesting SOX11 played an important role during the pathogenesis of OA. Thus, we hypothesized that SOX11 could be a potential target for the treatment of patients with OA.

Pullulanibacillus camelliae sp. nov., isolated from Pu'er tea.

A novel Gram-stain-positive, strictly aerobic, endospore-forming, rod-shaped bacterial strain 7578-24T was isolated from ripened Pu'er tea. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 7578-24T clustered with species of the genus Pullulanibacillus in the family Sporolactobacillaceae with 97.8-95.2 % sequence similarities, and was most closely related to Pullulanibacillus pueri YN3T with 97.8 % 16S rRNA gene sequence similarity. The DNA-DNA relatedness value between strain 7578-24T and P. pueri YN3T was 35 %. Strain 7578-24T had a cell-wall type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. The major menaquinone was menaquinone 7 (MK-7). C18 : 1ω7c (45.4 %), anteiso-C17 : 0 (30.6 %) and anteiso-C15 : 0 (10.1 %) were the predominant fatty acids, and diphosphatidylglycerol, phosphatidylglycerol, five unknown phospholipids and one unknown aminolipid were the major polar lipids. The DNA G+C content of strain 7578-24T was 45.2 mol%. Strain 7578-24T could be differentiated from other related species of the genus Pullulanibacillus based on phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of polyphasic evidence from this study, a novel species of the genus Pullulanibacillus named Pullulanibacillus camelliae sp. nov. is proposed, with strain 7578-24T (=CGMCC 1.15371T=JCM 31236T) as the type strain.

Current Status of Biological Therapies for the Treatment of Metastatic Melanoma.

Compared to early-stage melanoma when surgical excision is possible, metastatic disease continues to offer a much grimmer prognosis as traditional chemotherapy treatment regimens offer relatively little survival benefit. This has led to changes in treatment approaches over the preceding two decades as contemporary methods for the treatment of advanced or metastatic melanoma now involve a number of biological modalities, which include immunotherapeutic approaches, targeted therapies and epigenetic modification therapies. Clinically available immunotherapeutic agents include interleukin 2 (IL-2), as well as drugs targeting the important immune checkpoint molecules, such as cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1). The targeted therapeutic agents modulate specific pro-oncogenic mutations such as v-Raf murine sarcoma viral oncogene homolog B (BRAF), receptor tyrosine kinases, MEK inhibitors and potential future therapeutic targets, such as the CDK4/CDK6, PTEN and GNAQ/GNA11 genes. Additionally, an increasing understanding of the role of epigenetic alterations in the development and progression of melanoma now offers a new potential drug target. Several of these agents have shown promising results; however, in many investigations, combinations of different therapeutic approaches, each with different mechanisms of action, have yielded improved outcomes as treatment regimens continue to be further optimized by active research and patient disease sub-group analyses. This review summarizes the novel biological agents and new treatments, directly contributing to the significant improvement of biological therapies and markedly advancing knowledge of clinical application of newly approved and developed therapies in treatment of patients with metastatic melanoma.

Aeromicrobium camelliae sp. nov., isolated from Pu'er tea.

A novel Gram-reaction-positive, aerobic and non-spore-forming rod-shaped bacterial strain, YS17T, was isolated from ripened Pu'er tea. Growth of the strain was observed at 15-50 °C (optimum 30-37 °C) and at pH 5.5-10.5 (optimum 6.0-9.5). Phylogenetic analysis of 16S rRNA gene sequences indicated that the strain represented a member of the genus Aeromicrobium. The strains most closely related to YS17T were Aeromicrobium erythreum DSM 8599T, Aeromicrobium alkaliterrae JCM 13518T and Aeromicrobium ginsengisoli JCM 14732T, with 16S rRNA gene sequence similarities of 96.8, 96.8 and 96.7 %, respectively. DNA-DNA hybridization of YS17T with the type strains of the most closely related species, A. erythreum DSM 8599T, A. alkaliterrae JCM 13518T and A. ginsengisoli JCM 14732T, yielded reassociation values of 10.9, 16.8 and 10.9 %, respectively. The diagnostic diamino acid of the cell wall peptidoglycan was ll-diaminopimelic acid. The predominant menaquinones were menaquinone MK-9(H4) (76 %) and MK-8(H4) (17 %). The major fatty acids were C16 : 0, 10-methyl C18 : 0 and C18 : 1ω9c. The DNA G+C content of YS17T was 66 mol%. YS17T could be differentiated from recognized species of the genus Aeromicrobium on the basis of phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of evidence from the polyphasic analyses performed as part of this study a novel species, Aeromicrobium camelliae sp. nov., is proposed, with strain YS17T ( = CGMCC 1.12942T = JCM 30952T) as the type strain.

Pullulanibacillus pueri sp. nov., isolated from Pu'er tea.

A novel Gram-stain-positive, aerobic, endospore-forming, rod-shaped bacterial strain YN3(T) was isolated from ripened Pu'er tea. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain belonged to the family Sporolactobacillaceae and was closely related to Pullulanibacillus naganoensis DSM 10191(T) (95.8% 16S rRNA gene sequence similarity) and Pullulanibacillus uraniitolerans DSM 19429(T) (95.4%). Growth of the strain was observed at 20-50 °C (optimum 30-37 °C), at pH 4.0-8.0 (optimum pH 5.0-6.0). The strain had a cell-wall type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinone was menaquinone-7 (MK-7). The major fatty acids were anteiso-C15:0, anteiso-C17:0 and C18:1ω7c. The DNA G+C content of strain YN3(T) was 38.7 mol%. Strain YN3(T) could be differentiated from recognized species of the genus Pullulanibacillus based on phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of polyphasic evidence from this study, Pullulanibacilluspueri sp. nov., is proposed, with strain YN3(T) ( = CGMCC 1.12777(T ) = JCM 30075(T)) as the type strain.

Paenibacillus yunnanensis sp. nov., isolated from Pu'er tea.

A novel Gram-staining-positive, aerobic, endospore-forming, rod-shaped bacterial strain, YN2T, was isolated from ripened Pu'er tea. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain represented a novel species of the genus Paenibacillus. The strains most closely related to strain YN2T were Paenibacillus vulneris JCM 18268T and Paenibacillus rigui JCM 16352T, with 16S rRNA similarities of 98.6 and 95.5 %, respectively. Chemotaxonomic data supported the affiliation of the new isolate to the genus Paenibacillus, including MK-7 as the major menaquinone, DNA G+C content of 51 mol%, cell-wall type A1γ (meso-diaminopimelic acid as the diagnostic diamino acid) and anteiso-C15 : 0 and iso-C16 : 0 as the major fatty acids. Major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine and phospholipid. Strain YN2T could be differentiated from recognized species of the genus Paenibacillus based on phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of evidence from this polyphasic study, Paenibacillus yunnanensis sp. nov., is proposed, with strain YN2T ( = CGMCC 1.12968T = JCM 30953T) as the type strain.

Desnutrin/ATGL activates PPARδ to promote mitochondrial function for insulin secretion in islet ß cells.

Excessive caloric intake leading to obesity is associated with insulin resistance and dysfunction of islet ß cells. High-fat feeding decreases desnutrin (also called ATGL/PNPLA2) levels in islets. Here we show that desnutrin ablation via RIP-Cre (ßKO) or RIP-CreER results in hyperglycemia with impaired glucose-stimulated insulin secretion (GSIS). Due to decreased lipolysis, islets have higher TAG content but lower free FA levels. ßKO islets exhibit impaired mitochondrial respiration and lower production of ATP required for GSIS, along with decreased expression of PPARδ target genes involved in mitochondrial oxidation. Furthermore, synthetic PPARδ, but not PPARα, agonist restores GSIS and expression of mitochondrial oxidative genes in ßKO mice, revealing that desnutrin-catalyzed lipolysis generates PPARδ ligands. Finally, adenoviral expression of desnutrin in ßKO islets restores all defects of ßKO islet phenotype and function, including GSIS and mitochondrial defects, demonstrating the critical role of the desnutrin-PPARδ-mitochondrial oxidation axis in regulating islet ß cell GSIS.

Phosphorylation and recruitment of BAF60c in chromatin remodeling for lipogenesis in response to insulin.

Fatty acid and triglyceride synthesis is induced in response to feeding and insulin. This lipogenic induction involves coordinate transcriptional activation of lipogenic enzymes, including fatty acid synthase and glycerol-3-phosphate acyltransferase. We recently reported the importance of USF-1 phosphorylation and subsequent acetylation in insulin-induced lipogenic gene activation. Here, we show that Brg1/Brm-associated factor (BAF) 60c is a specific chromatin remodeling component for lipogenic gene transcription in liver. In response to insulin, BAF60c is phosphorylated at S247 by atypical PKCζ/λ, which causes translocation of BAF60c to the nucleus and allows a direct interaction of BAF60c with USF-1 that is phosphorylated by DNA-PK and acetylated by P/CAF. Thus, BAF60c is recruited to form the lipoBAF complex to remodel chromatin structure and to activate lipogenic genes. Consequently, BAF60c promotes lipogenesis in vivo and increases triglyceride levels, demonstrating its role in metabolic adaption to activate the lipogenic program in response to feeding and insulin.

Uncoupling of inflammation and insulin resistance by NF-kappaB in transgenic mice through elevated energy expenditure.

To study the metabolic activity of NF-kappaB, we investigated phenotypes of two different mouse models with elevated NF-kappaB activities. The transcriptional activity of NF-kappaB is enhanced either by overexpression of NF-kappaB p65 (RelA) in aP2-p65 mice or inactivation of NF-kappaB p50 (NF-kappaB1) through gene knock-out. In these models, energy expenditure was elevated in day and night time without a change in locomotion. The mice were resistant to adulthood obesity and diet-induced obesity without reduction in food intake. The adipose tissue growth and adipogenesis were inhibited by the elevated NF-kappaB activity. Peroxisome proliferator-activator receptor gamma expression was reduced by NF-kappaB at the transcriptional level. The two models exhibited elevated inflammatory cytokines (tumor necrosis factor-alpha and interleukin-6) in adipose tissue and serum. However, insulin sensitivity was not reduced by the inflammation in the mice on a chow diet. On a high fat diet, the mice were protected from insulin resistance. The glucose infusion rate was increased more than 30% in the hyperinsulinemic-euglycemic clamp test. Our data suggest that the transcription factor NF-kappaB promotes energy expenditure and inhibits adipose tissue growth. The two effects lead to prevention of adulthood obesity and dietary obesity. The energy expenditure may lead to disassociation of inflammation with insulin resistance. The study indicates that inflammation may prevent insulin resistance by eliminating lipid accumulation.