Inhibition of thioredoxin reductase and upregulation of apoptosis genes for effective anti-tumor sono-chemotherapy using a meso-organosilica nanomedicine.

The thioredoxin system is involved in cancer development and therefore is a promising target for cancer chemotherapy. Thioredoxin reductase (TrxR) is a key component of the thioredoxin (Trx) system, and is overexpressed in many cancers to inhibit apoptosis-related proteins. Alternatively, inhibition of thioredoxin reductase and upregulation of apoptosis factors provide a therapeutic strategy for anti-tumor treatment. In this study, an ultrasound-activatable meso-organosilica nanomedicine was prepared by integrating chloroquine (CQ) into hollow mesoporous organosilica (CQ@MOS). The meso-organosilica nanomedicine can inhibit the activity of thioredoxin reductase, elevate cellular reactive oxygen species (ROS) levels, upregulate the pro-apoptotic factors in the c-Jun N-terminal kinase (JNK) apoptosis pathway and induce autophagy inhibition, further resulting in mitochondrial membrane potential (MMP) depolarization and cellular ATP content decrease, ultimately causing significant damage to tumor cells. Moreover, CQ@MOS can efficiently deliver chloroquine into cancer cells and promote an enhanced sonodynamic effect for effective anti-tumor chemotherapy and sonodynamic therapy. This study may enlighten us on a new anti-tumor strategy and suggest its promising applications in cancer treatments.

Integrated organosilica nanomedicine enables sonoimaging, sonochemistry and antitumor sonodynamic therapy.

Sonography with its non-invasive and deep tissue-penetrating characteristics, not only contributes to promising developments in clinical disease diagnosis but also obtains acknowledgments as a prospective therapeutic approach in the field of tumor treatment. However, it remains a challenge for sonography simultaneously to achieve efficient imaging and therapeutic functionality. Here, we present an innovative integrated diagnosis and treatment paradigm by developing the nanomedicine of percarbamide-bromide-mesoporous organosilica spheres (MOS) with RGD peptide modification (PBMR) by loading percarbamide and bromide in MOS which were prepared by a one-step O/W microemulsion method. The PBMR nanomedicine effectively modifies the tumor acoustic environment to improve sonoimaging efficacy and induces sonochemical reactions to enhance the production of reactive oxygen species (ROS) for tumor treatment efficiency under sonography. The combination of PBMR nanomedicine and SDT achieved multiple ROS generation through the controlled sonochemical reactions and significantly boosted the potency of sonodynamic therapy and induced significant tumor regression with non-invasive tissue penetrability and minimizing damage to healthy tissues. Simultaneously, the generation of oxygen gas in the sonochemical process augments ultrasound reflection, resulting in a 4.9-fold increase in imaging grayscale. Our research establishes an effective platform for the synergistic integration of sonoimaging and sonodynamic antitumor therapy, offering a novel approach for precise antitumor treatment in the potential clinical applications.

Elucidating the Roles of Nafion/Solvent Formulations in Copper-Catalyzed CO2 Electrolysis.

Nafion ionomer, composed of hydrophobic perfluorocarbon backbones and hydrophilic sulfonic acid side chains, is the most widely used additive for preparing catalyst layers (CLs) for electrochemical CO2 reduction, but its impact on the performance of CO2 electrolysis remains poorly understood. Here, we systematically investigate the role of the catalyst ink formulation on CO2 electrolysis using commercial CuO nanoparticles as the model pre-catalyst. We find that the presence of Nafion is essential for achieving stable product distributions due to its ability to stabilize the catalyst morphology under reaction conditions. Moreover, the Nafion content and solvent composition (water/alcohol fraction) regulate the internal structure of Nafion coatings, as well as the catalyst morphology, thereby significantly impacting CO2 electrolysis performance, resulting in variations of C2+ product Faradaic efficiency (FE) by >3×, with C2+ FE ranging from 17 to 54% on carbon paper substrates. Using a combination of ellipsometry and in situ Raman spectroscopy during CO2 reduction, we find that such selectivity differences stem from changes to the local reaction microenvironment. In particular, the combination of high water/alcohol ratios and low Nafion fractions in the catalyst ink results in stable and favorable microenvironments, increasing the local CO2/H2O concentration ratio and promoting high CO surface coverage to facilitate C2+ production in long-term CO2 electrolysis. Therefore, this work provides insights into the critical role of Nafion binders and underlines the importance of optimizing Nafion/solvent formulations as a means of enhancing the performance of electrochemical CO2 reduction systems.

Perioperative Electroacupuncture Can Accelerate the Recovery of Gastrointestinal Function in Cancer Patients Undergoing Pancreatectomy or Gastrectomy: A Randomized Controlled Trial.

The effect of perioperative acupuncture on accelerating gastrointestinal function recovery has been reported in colorectal surgery and distal gastrectomy (Billroth-II). However, the evidence in pancreatectomy and other gastrectomy is still limited. A prospective, randomized controlled trial was conducted between May 2018 and August 2019. Consecutive patients undergoing pancreatectomy or gastrectomy in our hospital were randomly assigned to the electroacupuncture (EA) group and the control group. The patients in the EA group received transcutaneous EA on Bai-hui (GV20), Nei-guan (PC6), Tian-shu (ST25), and Zu-san-li (ST36) once a day in the afternoon, and the control group received sham EA. Primary outcomes were the time to first flatus and time to first defecation. In total, 461 patients were randomly assigned to the groups, and 385 were analyzed finally (EA group, n = 201; control group, n = 184). Time to first flatus (3.0 ± 0.7 vs 4.2 ± 1.0, P < 0.001) and first defecation (4.2 ± 0.9 vs 5.4 ± 1.2, P < 0.001) in the EA group were significantly shorter than those in the control group. Of patients undergoing pancreatectomy, those undergoing pancreaticoduodenectomy and intraoperative radiation therapy (IORT) surgery benefitted from EA in time to first flatus (P < 0.001) and first defecation (P < 0.001), while those undergoing distal pancreatectomy did not (P flatus=0.157, P defecation=0.007) completely. Of patients undergoing gastrectomy, those undergoing total gastrectomy and distal gastrectomy (Billroth-II) benefitted from EA (P < 0.001), as did those undergoing proximal gastrectomy (P=0.015). Patients undergoing distal gastrectomy (Billroth-I) benefitted from EA in time to first defecation (P=0.012) but not flatus (P=0.051). The time of parenteral nutrition, hospital stay, and time to first independent walk in the EA group were shorter than those in the control group. No severe EA complications were reported. EA was safe and effective in accelerating postoperative gastrointestinal function recovery. Patients undergoing pancreaticoduodenectomy, IORT surgery, total gastrectomy, proximal gastrectomy, or distal gastrectomy (Billroth-II) could benefit from EA. This trial is registered with NCT03291574.

Suppression of the long non-coding RNA MALAT-1 impairs the growth and migration of human tongue squamous cell carcinoma SCC4 cells.

INTRODUCTION: Aberrant expression of long non-coding RNAs (lncRNAs) is associated with metastasis and poor prognosis in patients with various cancer types. However, few studies have assessed lncRNAs in oral squamous cell carcinoma (OSCC). This study aimed to investigate the expression and impact of lncRNAs in OSCC. MATERIAL AND METHODS: Real-time PCR analysis was used to examine the expression of four lncRNAs, MALAT-1, UCA1, BC200 and SRA, in 14 OSCC and adjacent normal tissue pairs. The impact of MALAT-1 suppression by siRNA on the proliferation, apoptosis, anchorage-independent growth and migration of the human tongue carcinoma cell line SSC4 was also determined. RESULTS: MALAT-1 levels were significantly higher in the OSCC tissue than in the normal tissues (p < 0.004); no significant differences in UCA1, BC200 or SRA RNA levels were observed. Knockdown of MALAT-1 by siRNA significantly suppressed proliferation of SSC4 cells (p < 0.004) and enhanced their apoptosis (p < 0.001). In addition, siRNA-mediated suppression of MALAT-1 inhibited SSC4 cell colony formation (p < 0.001) and migration (p < 0.004). CONCLUSIONS: Elevated expression of MALAT-1 in OSCC may play a role in tumorigenesis and/or metastasis. Further studies are necessary to identify the mechanism by which MALAT-1 influences SCC4 growth and migration and validate its increased expression in OSCC patients.

Construction of prognostic risk prediction model of oral squamous cell carcinoma based on co-methylated genes.

This study aimed to identify DNA methylation markers in oral squamous cell carcinoma (OSCC) and to construct a prognostic prediction model of OSCC. For this purpose, the methylation data of patients with OSCC downloaded from The Cancer Genome Atlas were considered as a training dataset. The methylation profiles of GSE37745 for OSCC samples were downloaded from Gene Expression Omnibus and considered as validation dataset. Differentially methylated genes (DMGs) were screened from the TCGA training dataset, followed by co­methylation analysis using weighted correlation network analysis (WGCNA). Subsequently, the methylation and gene expression levels of DMGs involved in key modules were extracted for correlation analysis. Prognosis­related methylated genes were screened using the univariate Cox regression analysis. Finally, the risk prediction model was constructed and validated through GSE52793. The results revealed that a total of 948 DMGs with CpGs were screened out. Co­methylation gene analysis obtained 2 (brown and turquoise) modules involving 380 DMGs. Correlation analysis revealed that the methylation levels of 132 genes negatively correlated with the gene expression levels. By combining with the clinical survival prognosis of samples, 5 optimized prognostic genes [centromere protein V (CENPV), Tubby bipartite transcription factor (TUB), synaptotagmin like 2 (SYTL2), occludin (OCLN) and CAS1 domain containing 1 (CASD1)] were selected for constructing a risk prediction model. It was consistent in the training dataset and GSE52793 that low­risk samples had a better survival prognosis. On the whole, this study indicates that the constructed risk prediction model based on CENPV, SYTL2, OCLN, CASD1, and TUB may have the potential to be used for predicting the survival prognosis of patients with OSCC.

Twenty­four signature genes predict the prognosis of oral squamous cell carcinoma with high accuracy and repeatability.

Oral squamous cell carcinoma (OSCC) is the sixth most common type cancer worldwide, with poor prognosis. The present study aimed to identify gene signatures that could classify OSCC and predict prognosis in different stages. A training data set (GSE41613) and two validation data sets (GSE42743 and GSE26549) were acquired from the online Gene Expression Omnibus database. In the training data set, patients were classified based on the tumor­node­metastasis staging system, and subsequently grouped into low stage (L) or high stage (H). Signature genes between L and H stages were selected by disparity index analysis, and classification was performed by the expression of these signature genes. The established classification was compared with the L and H classification, and fivefold cross validation was used to evaluate the stability. Enrichment analysis for the signature genes was implemented by the Database for Annotation, Visualization and Integration Discovery. Two validation data sets were used to determine the precise of classification. Survival analysis was conducted followed each classification using the package 'survival' in R software. A set of 24 signature genes was identified based on the classification model with the Fi value of 0.47, which was used to distinguish OSCC samples in two different stages. Overall survival of patients in the H stage was higher than those in the L stage. Signature genes were primarily enriched in 'ether lipid metabolism' pathway and biological processes such as 'positive regulation of adaptive immune response' and 'apoptotic cell clearance'. The results provided a novel 24­gene set that may be used as biomarkers to predict OSCC prognosis with high accuracy, which may be used to determine an appropriate treatment program for patients with OSCC in addition to the traditional evaluation index.

Adrenomedullin promotes the odontogenic differentiation of dental pulp stem cells through CREB/BMP2 signaling pathway.

Adrenomedullin (AM) could promote the proliferation, the odontogenic differentiation and inhibit the apoptosis of dental pulp stem cells (DPSCs). AM in combination with DPSCs may be an effective strategy for pulp repair. However, there was no report on the mechanisms of AM in the odontogenic differentiation of DPSCs. The aim of this study is to investigate the molecular mechanisms through which AM promotes the odontogenic differentiation of DPSCs. Freshly extracted wisdom teeth were obtained from 27 patients. Cells at passage 3 to passage 5 were used in this study. DPSCs were treated with or without 10-7 M AM in Dulbecco's modified Eagle's medium culture, and then the accumulated calcium deposition was analyzed after 21 days by using alizarin red S staining. Odontogenic differentiation markers were determined by western blot analysis and quantitative real-time PCR. Western blot analysis results showed that AM had the capability of promoting the odontogenic differentiation of DPSCs and AM could enhance the phosphorylation of CREB and up-regulate the expression of BMP2. H89 is a CREB inhibitor which can inhibit the odontogenic differentiation of DPSCs through inhibiting the phosphorylation of CREB. Noggin could inhibit the odontogenic differentiation of DPSCs through inhibiting the activity of BMP2. These results indicated that AM could promote the odontogenic differentiation of DPSCs by upregulating the expression of BMP2 through the CREB signaling pathway.

Interaction between MALAT-1, CCR7 and correlated genes in oral squamous cell carcinoma.

OBJECTIVE: This study focuses on the feasible molecular mechanism of the interaction between MALAT-1, CCR7 and related genes in oral squamous cell carcinoma, to find new target molecules that can block the lymph node metastasis. METHODS: The expression of MALAT-1, miRNA-320s, SRSF1, YB-1 and CCR7 were detected in T3/T4-phase OSCC tissues of two groups with or without lymph node metastasis using real-time qPCR. CO-IP and western blot to test the interaction of RNAs (MALAT-1, miRNA-320s) with SRSF1 protein or YB-1 were evaluated by CO-IP, Western blot and real-time qPCR. The expression change of chemokine receptor CCR7 were investigated using CO-IP, Western blot and real-time qPCR after silencing miRNA-320d (one of the miRNA-320s family members) by transfection of miRNA mimics to explore related signaling pathway. RESULTS: The expression levels of MALAT-1 SRSF1 and CCR7 in OSCC tissues with were differentially higher compared with those of samples without lymph node metastasis as well as para-carcinoma tissues, exclusive of miRNA-320d. Moreover, it is confirmed that the target RNA (MALAT-1, miRNA-320s) and SRSF1 protein can combine with each other, based on the statistically significant difference compared with negative control group (P<0.05). In addition, the expression of CCR7 was higher than the negative control group after silencing miRNA-320d. CONCLUSION: SRSF1 is likely to mediate the interactive relationship between MALAT-1 and miRNA-320d. CCR7 expression can be distinctly increased by silencing miRNA-320d. The effect of long-chain non-coding RNA MALAT-1 on chemokine receptor CCR7 and possibly further influence on lymph node metastasis of oral squamous cell carcinoma are revealed in molecular level to offer help for prevention and treatment of OSCC in future.

Comparative Genomic Analysis of Delftia tsuruhatensis MTQ3 and the Identification of Functional NRPS Genes for Siderophore Production.

Plant growth-promoting rhizobacteria (PGPR) are a group of rhizosphere bacteria that promote plant growth. Delftia tsuruhatensis MTQ3 is a member of PGPR that produces siderophores. The draft genome sequence of MTQ3 has been reported. Here, we analyzed the genome sequence of MTQ3 and performed a comparative genome analysis of four sequenced Delftia strains, revealing genetic relationships among these strains. In addition, genes responsible for bacteriocin and nonribosomal peptide synthesis were detected in the genomes of each strain. To reveal the functions of NRPS genes in siderophore production in D. tsuruhatensis MTQ3, three NRPS genes were knocked out to obtain the three mutants MTQ3-Δ1941, MTQ3-Δ1945, and MTQ3-Δ1946, which were compared with the wild-type strain. In qualitative and quantitative analyses using CAS assay, the mutants failed to produce siderophores. Accordingly, the NRPS genes in MTQ3 were functionally related to siderophore production. These results clarify one mechanism by which plant growth is promoted in MTQ3 and have important applications in agricultural production.

Adrenomedullin Promotes the Proliferation and Inhibits Apoptosis of Dental Pulp Stem Cells Involved in Divergence Pathways.

INTRODUCTION: Adrenomedullin (ADM) is highly expressed in dental tissues at the critical developmental time points during tooth development. However, its role in pulp repair and pulp injury is still unknown. The study aimed to investigate the mechanisms by which ADM affects the proliferation and apoptosis of human dental pulp stem cells (DPSCs). METHODS: Fifty-four freshly extracted wisdom teeth were obtained from 27 patients with caries and 27 without. Enzyme-linked immunosorbent assay and immunohistochemistry were used to examine the expression levels of ADM in caries and normal pulp tissues. Human primary DPSCs from the third passage were incubated with ADM (10(-8) mol/L) for 24 hours; the cell cycle and apoptosis were investigated by flow cytomery, and protein expression was investigated using Western blot analysis. RESULTS: The levels of ADM in caries pulp tissues were significantly higher than those in normal pulp tissues. Incubation with ADM enhanced the percentage of G2/S/M phase DPSCs (P < .01) by the addition of a JNK/c-Jun inhibitor. Incubation with ADM reduced DPSC apoptosis by the addition of the Src/glycogen synthase kinase-3 inhibitor. ADM also ameliorated CoCl2-induced apoptosis and caspase-3 expression. CONCLUSIONS: ADM enhances the proliferation of DPSCs through activation of the JNK/c-Jun signaling pathway and inhibits apoptosis of DPSCs through activation of the Src/glycogen synthase kinase-3 signaling pathway.

Synthesis of trace element Si and Sr codoping hydroxyapatite with non-cytotoxicity and enhanced cell proliferation and differentiation.

The main inorganic minerals in natural bones are non-stoichiometric hydroxyapatite (HA, Ca10[PO4]6[OH]2) doped with various trace elements, which may possess important biochemical effects. To investigate the functions of Sr and Si elements in human hard tissues, non-doped HA, trace Si doped HA, Si and Sr codoped HA with the concentration of natural bones are synthesized by hydrothermal method in this study. The samples are characterized by powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). The biological activities are evaluated via cytotoxicity study, adhesion and proliferation of osteoblast measurement, and alkaline phosphatase (ALP) assay. All the synthesized materials are HA phase, which have hierarchical structures with oriented HA nanorods assembled into the platy particles. These materials are non-cytotoxic against L929 cells line even at 400 µg/ml powder suspension. The results clearly indicate that the proliferation of L929 cells increases with trace elements doping from trace Si-HA to Si + Sr-HA. The adhesion and proliferation of osteoblast measurement illustrates that proliferation of osteoblasts advances about 1.3 times for Si-HA and about 1.8 times for Si + Sr-HA compared with undoped HA. In general, Si-HA with trace Si element shows enhanced cell differentiation, and Si + Sr-HA dual-doped with Si and Sr elements presents increased biological activity compared with Si-HA.

Functional hydroxyapatite bioceramics with excellent osteoconductivity and stern-interface induced antibacterial ability.

The biocompatibility and antibacterial properties of hydroxyapatite (HAp) bioceramics are crucial in medical applications. However, it is still a challenge to control HAp with antibacterial ability while maintaining other biological properties in the development of bioactive bone implants. Herein, we report functional silver ion substituted HAp bioceramics with excellent osteoconductivity and efficient antibacterial activity and propose a stern-interface induced antibacterial mechanism of such bioactive ceramics. In this antibacterial process, the concentration of Ag(+) at the stern-interface of Ag/HAp bioceramics is nearly 5 times higher than that in the bulk solution due to the trace dopant Ag(+) enrichment in the stern layer of the electric double layer at the negatively charged surface of Ag/HAp bioceramics. Trace Ag-doping in HAp induces a positive shift of zeta potential and increase of hydrophilicity, which may help inhibit bacterial proliferation. The positive osteoblast adhesion, proliferation and differentiation of ultra-trace doped Ag/HAp are also demonstrated through actin cytoskeleton staining, MTT and alkaline phosphatase (ALP) activity assays. This work may enlighten us on the artificial design of novel smart anti-infective bone grafts using ultra-trace functional elements and also suggest its potential applications in orthopedic surgery and bone osseointegration.

Ultra-trace silver-doped hydroxyapatite with non-cytotoxicity and effective antibacterial activity.

Hydroxyapatite (HAp) nanocrystals as the main inorganic component in hard tissue have been extensively studied for bone regeneration and dental implant treatment. However, failure of surgical reconstruction often occurs owing to the lack of effective antibacterial ability of HAp. It is still a challenge to develop artificial HAp with both efficient antibacterial ability and proper biological properties. Herein, a series of ultra-trace Ag-doped HAp nanocrystals have been elaborately prepared with the optimal doping concentration from 0.27 ppm to 2.2 ppm, which present non-cytotoxicity while possess effective bacteria reduction ability. Ultra-trace Ag-doped HAp nanocrystals possess higher protein adsorption than pure HAp nanocrystals due to the trace doping-induced less negative surface potential. The ultra-trace Ag-doped HAp nanocrystals showed effectively antibacterial ability, non-cytotoxicity and enhanced adsorbability that made them ideal materials for various biocompatible and antibacterial applications.

High genetic diversity and novelty in eukaryotic plankton assemblages inhabiting saline lakes in the Qaidam basin.

Saline lakes are intriguing ecosystems harboring extremely productive microbial communities in spite of their extreme environmental conditions. We performed a comprehensive analysis of the genetic diversity (18S rRNA gene) of the planktonic microbial eukaryotes (nano- and picoeukaryotes) in six different inland saline lakes located in the Qaidam Basin. The novelty level are high, with about 11.23% of the whole dataset showing <90% identity to any previously reported sequence in GenBank. At least 4 operational taxonomic units (OTUs) in mesosaline lakes, while up to eighteen OTUs in hypersaline lakes show very low CCM and CEM scores, indicating that these sequences are highly distantly related to any existing sequence. Most of the 18S rRNA gene sequence reads obtained in investigated mesosaline lakes is closely related to Holozoa group (48.13%), whereas Stramenopiles (26.65%) and Alveolates (10.84%) are the next most common groups. Hypersaline lakes in the Qaidam Basin are also dominated by Holozoa group, accounting for 26.65% of the total number of sequence reads. Notably, Chlorophyta group are only found in high abundance in Lake Gasikule (28.00%), whereas less represented in other hypersaline lakes such as Gahai (0.50%) and Xiaochaidan (1.15%). Further analysis show that the compositions of planktonic eukaryotic assemblages are also most variable between different sampling sites in the same lake. Out of the parameters, four show significant correlation to this CCA: altitude, calcium, sodium and potassium concentrations. Overall, this study shows important gaps in the current knowledge about planktonic microbial eukaryotes inhabiting Qaidam Basin (hyper) saline water bodies. The identified diversity and novelty patterns among eukaryotic plankton assemblages in saline lake are of great importance for understanding and interpreting their ecology and evolution.