Increased ten-eleven translocation methylcytosine dioxygenase one in dorsal root ganglion contributes to inflammatory pain in CFA rats.

DNA hydroxylation catalyzed by Tet dioxygenases occurs abundantly in neurons in mammals. However, effects of ten-eleven translocation methylcytosine dioxygenase 1 (TET1) expression and hydroxymethylation status on neuron injury remain unclear. This study was designed to explore the effects of TET1 and TET2 expression in the inflammatory pain of rats induced by complete Freund's adjuvant (CFA). Mechanical paw withdrawal threshold (PWT) and thermal withdrawal latency (TWL) were detected to assess pain behavior. The expression of TET1 and TET2 were measured in the dorsal root ganglion (DRG) with western blotting analysis. Immunofluorescence staining is employed to detect the expression and co-location of TRPV1 with TET1. Intrathecal administration of Bobcat339 was used to inhibit TET1 function in dorsal root ganglion. The paw withdrawal threshold and thermal withdrawal latency of rats were significantly reduced after CFA Injection. Western blot results showed that the expression of TET1 was significantly increased at 3 days after CFA injection, but TET2 had no statistical difference. Immunofluorescence results showed that TET1 was co-localized with TRPV1. Intrathecal administration of Bobcat339 improved mechanical and thermal pain threshold in CFA rats. Our findings highlight the role of TET1 in chronic inflammatory pain model. The expression of TET1 was increased in CFA rats, and suppression of TET1 will ameliorate inflammatory pain.

[Construction of protoplast genetic transformation system for Mycena--symbiont of Gastrodia elata].

Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.

Electro-optical performance of polymer-stabilized sphere phase liquid crystal displays.

Electro-optical properties of polymer-stabilized sphere phase liquid crystal (PS-SPLC) switching devices are analyzed and validated experimentally. The experimental results show the voltage-dependent transmittance curves of PS-SPLC devices. A diffraction approach, called extended anomalous diffraction approach, is proposed to fit the experimental data. Good agreement between experiment and model is obtained. The scattering model provides practical guidance for the improvement of PS-SPLC displays performance and optimization.

Two new menthane monoterpenes from Illicium wardii.

Two new menthane monoterpenes, wardicines A (1) and B (2), were isolated from the fruits of Illicium wardii A.C. Smith. Their structures were determined on the basis of spectroscopic data analysis. Cytotoxicities of compounds 1 and 2 against three human tumor cell lines A549, HCT-116, and CCRF-CEM were also evaluated, but no significant activities were observed for the two compounds.

Two novel lignans from Gaultheria yunnanensis.

Two new lignans, 9-salicyl-(+)-isolariciresinol (1) and gaultheroside G (2), together with seven known compounds, were isolated from the ethanolic extract of the whole plant of Gaultheria yunnanensis. Their structures were determined by extensive NMR and MS analyses. Gaultheroside G (2) was found to have an unusual ether linkage between the 2 and 9' positions of aryl-tetralin lignan skeleton. All nine compounds were assayed for inhibitory effects against nitric oxide and pro-inflammatory cytokines TNF-α and IL-6 release in LPS-induced RAW 246.7 macrophages, while no significant activities were observed for the evaluated compounds.

Anti-rheumatoid arthritis potential of different fractions derived from of Coluria longifolia.

Coluria longifolia Maxim (C. longifolia) is a Chinese folk medication commonly used to treat arthritis and joint pain. Literatures have reported that C. longifolia has significant anti-inflammatory, analgesic and antipyretic effects. The aim of this research was to assay the effective fractions of C. longifolia (EFCL) against rheumatoid arthritis (RA) and to elucidate its anti-RA mechanism on a preliminary basis. The rat model of collagen-induced arthritis (CIA) was established. The therapeutic effects of different fractions in vivo were evaluated by body weight changes, a foot swelling score, inflammatory factors and histopathological examination. The mechanism of EFCL was investigated by activity of oxidative stress related enzyme, qPCR and Western blotting tests. In vivo results showed that total extraction (TE) and n-butanol fraction (NF) could significantly alleviate the symptoms of RA, decrease the levels of IL-6 and TNF-α (P < 0.01), and improve histopathological injury. The mechanism study showed that SOD level was significantly increased with MDA level decreased in the NF group. The upregulated proteins and mRNA expression levels of Nrf2, HO1 and NQO1 after TE and NF administration suggested that the anti-arthritic effect may be related to the Nrf2 signaling pathway and downstream HO1 and NQO1. In conclusion, this study confirmed that C. longifolia is capable of treating RA with NF as the main effective fraction. Its anti-RA action may be associated with Nrf2 signaling pathway and downstream HO1 and NQO1.

Trichostatin A relieves anxiety-and depression-like symptoms in APP/PS1 mice.

Background: Cognitive deficits and behavioral disorders such as anxiety and depression are common manifestations of Alzheimer's disease (AD). Our previous work demonstrated that Trichostatin A (TSA) could alleviate neuroinflammatory plaques and improve cognitive disorders. AD, anxiety, and depression are all associated with microglial inflammation. However, whether TSA could attenuate anxiety- and depression-like behaviors in APP/PS1 mice through anti-inflammatory signaling is still unclearly. Methods: In the present study, all mice were subjected to the open field, elevated plus maze, and forced swim tests to assess anxiety- and depression-related behaviors after TSA administration. To understand the possible mechanisms underlying the behavioral effects observed, CST7 was measured in the hippocampus of mice and LPS-treated BV2 microglia. Results: The results of this study indicated that TSA administration relieved the behaviors of depression and anxiety in APP/PS1 mice, and decreased CST7 levels in the hippocampus of APP/PS1 mice and LPS-induced BV2 cells. Conclusion: Overall, these findings support the idea that TSA might be beneficial for reducing neurobehavioral disorders in AD and this could be due to suppression of CST7-related microglial inflammation.

Overexpression of YTHDF3 increases the specific productivity of the recombinant protein in CHO cells by promoting the translation process.

Due to their high-quality characteristics, Chinese hamster ovary (CHO) cells have become the most widely used and reliable host cells for the production of recombinant therapeutic proteins in the biomedical field. Previous studies have shown that the m6A reader YTHDF3, which contains the YTH domain, can affect a variety of biological processes by regulating the translation and stability of target mRNAs. This study investigates the effect of YTHDF3 on transgenic CHO cells. The results indicate that stable overexpression of YTHDF3 significantly enhances recombinant protein expression without affecting host cell growth. Transcriptome sequencing indicated that several genes, including translation initiation factor, translation extension factor, and ribosome assembly factor, were upregulated in CHO cells overexpressing YTHDF3. In addition, cycloheximide experiments confirmed that YTHDF3 enhanced transgene expression by promoting translation in CHO cells. In conclusion, the findings in this study provide a novel approach for mammalian cell engineering to increase protein productivity by regulating m6A.

Micro-223 Promotes Diabetic Osteoarthritis Progression by Regulating Cartilage Degeneration and Subchondral Bone Remodeling.

OBJECTIVE: Our study was performed to investigate whether micro-223 promotes diabetic Osteoarthritis (OA) progression by regulating cartilage degeneration and subchondral bone remodeling. METHODS: The expression of miR-223 in human normal cartilage, OA cartilage, and subchondral bone tissue with or without DM was detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). miR-223 mimic or inhibitor was transfected into chondrocytes. Cell viability and apoptosis were assessed by 3-(4,5)-dimethylthiahiazo(-2)-3,5-diphenyltetrazolium bromide (MTT) and Terminal Deoxynucleotidyl Transferase(TdT)-mediated dUTP nick end labeling (TUNEL) assay, respectively. RESULTS: miR-223 was significantly higher in human diabetic OA cartilage and subchondral bone compared with normal OA and healthy control. Overexpression of miR-223 accelerated cartilage degeneration and subchondral bone sclerosis in diabetic OA mice, whereas miR-223 inhibition had the opposite effect. In vitro upregulation of miR-223 decreased proliferation and enhanced apoptosis of chondrocytes. Meanwhile, downregulation of miR-223 promoted glycosaminoglycan (GAG) production in chondrocytes. CONCLUSION: miR-223 promotes diabetic OA progression by regulating cartilage degeneration and subchondral bone remodeling both in vitro and in vivo.

Establishment and metabonomics analysis of nonalcoholic fatty liver disease model in golden hamster.

The aim is to establish a model of nonalcoholic fatty liver disease (NAFLD) caused by feeding with high-fat, high-fructose, and high-cholesterol diet (HFFCD) in golden hamsters, and to investigate the characteristics of the NAFLD model and metabolite changes of liver tissue. Golden hamsters were fed HFFCD or control diets for six weeks. Body weight, abdominal fat index, and liver index was assessed, serum parameters, hepatic histology, and liver metabolites were examined. The results showed that body weight, abdominal fat, and liver index of hamsters were significantly increased in the model group, the level of serum total cholesterol (TC), triglyceride (TG), and low density lipoprotein-cholesterol (LDL-C) were significantly increased in model group as well, and high density lipoprotein-cholesterol (HDL-C) was significantly decreased. In addition, lipid deposition in liver tissue formed fat vacuoles of different sizes. Metabonomics analysis of the liver showed that the metabolic pathways of sphingolipid, glycerophospholipids, and arginine biosynthesis were disordered in the NAFLD model. The modeling method is simple, short time, and uniform. It can simulate the early fatty liver caused by common dietary factors, and provides an ideal model for the study of the initial pathogenesis and therapeutic drugs for NAFLD.

Dataset for liver metabolomic profile of highland barley Monascus purpureus went extract-treated golden hamsters with nonalcoholic fatty liver disease.

Nonalcoholic Fatty Liver Disease (NAFLD) is a serious problem endangering human health in the world. The pathogenesis of this disease is often accompanied by lipid metabolism disorder and can cause liver lipid accumulation. Highland barley Monascus purpureus Went extract (HBMPWE) can inhibit the liver lipid accumulation caused by a high-fat, high-fructose, high-cholesterol diet. However, it is not clear what changes have taken place in the process of liver lipid metabolism after HBMPWE administration. To fill this knowledge gap and to support the findings published in the companion research article entitled "Highland Barley Monascus purpureus Went Extract Ameliorates High-Fat, High-Fructose, High-Cholesterol Diet Induced Nonalcoholic Fatty Liver Disease by Regulating Lipid Metabolism in Golden Hamsters" [1], we provided important information related to the liver differential metabolites and identified twenty-one differential metabolites of liver metabolism. In the model group, the levels of lactate, linoleic acid, and malic acid increased significantly. After HBMPWE treatment, the expressions of these metabolites reduced significantly. Therefore, these liver differential metabolites could be used as biological signatures reflecting the severity of NAFLD and HBMPWE treatment outcomes.

Symbiotic bacteria attenuate Drosophila oviposition repellence to alkaline through acidification.

Metazoans harbor a wealth of symbionts that are ever-changing the environment by taking up resources and/or excreting metabolites. One such common environmental modification is a change in pH. Conventional wisdom holds that symbionts facilitate the survival and production of their hosts in the wild, but this notion lacks empirical evidence. Here, we report that symbiotic bacteria in the genus Enterococcus attenuate the oviposition avoidance of alkaline environments in Drosophila. We studied the effects of alkalinity on oviposition preference for the first time, and found that flies are robustly disinclined to oviposit on alkali-containing substrates. This innate repulsion to alkaline environments is explained, in part, by the fact that alkalinity compromises the health and lifespan of both offspring and parent Drosophila. Enterococcus dramatically diminished or even completely reversed the ovipositional avoidance of alkalinity in Drosophila. Mechanistically, Enterococcus generate abundant lactate during fermentation, which neutralizes the residual alkali in an egg-laying substrate. In conclusion, Enterococcus protects Drosophila from alkali stress by acidifying the ovipositional substrate, and ultimately improves the fitness of the Drosophila population. Our results demonstrate that symbionts are profound factors in the Drosophila ovipositional decision, and extend our understanding of the intimate interactions between Drosophila and their symbionts.

Highly oxygenated guaianolides from Artemisia dubia.

Investigation of a Chinese folk medicine, Artemisia dubia (syn. Artemisia subdigitata), has resulted in the isolation of eight new guaianolides (artemdubolides A-H, 1-8), and nineteen known sesquiterpene lactones. Their structures were elucidated primarily on the basis of spectroscopic methods, including extensive 2D NMR experiments. The cytotoxic activities of compounds 1-8 were evaluated against A549 (human lung adenocarcinoma), Colo205 (human colon carcinoma), HepG2 (human hepatoma), and MDA-MB-435 (human melanoma) cell lines. Compounds 2 and 5 showed weak cell growth inhibition of Colo205 and MDA-MB-435 cells.

miR-21 may Act as a Potential Mediator Between Inflammation and Abnormal Bone Formation in Ankylosing Spondylitis Based on TNF-α Concentration-Dependent Manner Through the JAK2/STAT3 Pathway.

OBJECTIVE: To explore the role of microRNA (miR-21) in new bone formation in ankylosing spondylitis (AS) as mediated by different concentration of tumor necrosis factor-α (TNF-α). METHODS: Fibroblasts isolated from the hips of patients with AS were induced to osteogenesis. These cells were then stimulated with varying concentrations of TNF-α. MicroRNA-21 expressions were evaluated using reverse transcription-polymerase chain reaction (RT-PCR) and osteogenesis was detected via Alizarin Red S (ARS) staining and measurement of alkaline phosphatase (ALP) activity. Relative expressions of p-STAT3, Nuclear STAT3, cytoplasm STAT3, Runx2, BMP2, osteopontin, osteocalcin, and LC3B in AS fibroblasts were measured after exposure to different concentrations of TNF-α. The STAT3-inhibiting small interfering RNA allowed further exploration on its impact on miR-21 and primary miR-21 expressions. A proteoglycan-induced arthritis (PGIA) Balb/c mouse model was established in order to monitor sacroiliac joint (SIJ) inflammation and subsequent damage through magnetic resonance image. Serum miR-21 and TNF-α expressions were evaluated using RT-PCR and enzyme-linked immunosorbent assay. At week 16, mice models were transfected intravenously with miR-21 overexpressing agomir and miR-21 inhibiting antagomir for 7 successive days. The rate of abnormal bone formation at SIJ was evaluated using microcomputed tomography and hematoxylin and eosin staining at week 24. Western blot analysis enabled quantification of STAT-3, JAK-2, and interleukin (IL)-17A expressions present in the SIJ. RESULTS: The in vitro miR-21 expression and osteogenesis activity were noted to be augmented in the setting of low TNF-α concentrations (0.01-0.1 ng/mL) while they were depressed in settings with higher TNF-α concentrations (1-10 ng/mL). Samples with the most distinct ARS manifestation and ALP activity as well as the highest miR-21 expressions were those who received 0.1 ng/mL of TNF-α. Primary miR-21 was found to be notable raised by Si-STAT3, while the converse effect was seen in mature miR-21 expressions. Intravenous injection of exogenous miR-21 contributed to new bone formation and significantly elevated expressions of STAT3, JAK2, and IL-17 in PGIA mice. CONCLUSIONS: The results revealed that miR-21 may act as a potential mediator between new bone formation and inflammation in AS.

Serum miR-21 expression correlates with radiographic progression but also low bone mineral density in patients with ankylosing spondylitis: a cross-sectional study.

Increased expressions of miR-21 have been detected in ankylosing spondylitis (AS) patients. The current study was performed to examine the serum miR-21 expression with radiographic severity in AS patients, which was determined based on the modified New York (NY) criteria for sacroiliac joints assessment and modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS) system for spine involvement. Bone mineral density at lumbar 1-4 and femoral neck were examined by dual-energy absorptiometry (DXA). Serum miR-21 expressions were determined by quantitative real-time PCR, and receiver operating characteristic curve analysis was performed to identify the diagnostic value of miR-21 expression levels regarding the NY criteria. Elevated levels of serum miR-21 expressions were detected in AS patients compared with healthy controls. AS patients with modified NY grade 4 showed significantly higher miR-21 expression than grade 3 and grade 2. AS patients with spinal syndesmophytes had significantly higher serum miR-21 expressions than non-syndesmophyte patients. Increased miR-21 expressions were significantly related to the disease radiographic severity. In addition, serum miR-21 expressions were negatively associated with lumbar 1-4 and femoral neck bone mineral density. In summary, serum miR-21 expressions were related to structural damage and radiological progression in AS, indicating that miR-21 may act as a switch between inflammation and new bone information and regulate different signal ways between lesioned enthesis and trabecular bone.

[Relationship between RAR-beta gene expression defect and its methylation].

OBJECTIVE: To evaluate the expression of RAR-beta gene in cervical carcinoma cell lines SiHa, HeLa, C33A and Caski and to analyze the relation between their gene expression and the promoter methylation of RAR-beta DNA. METHODS: The expression of mRNA and protein of RAR-beta gene in the four cell lines were analyzed by RT-PCR, western blot and immunofluorescence, respectively. Methylation specific PCR (MSP) was used to check whether there was methylation in the promoter of RAR-beta gene. The demethylating agent 5-aza-2'-deoxycytidine (5-Aza-cdR) was used to treat methylated cell lines and the change of RAR-beta gene methylation and RAR-beta gene expression defects were observed. The cell proliferation was assayed with 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide method. RESULTS: The mRNA and protein expression levels of RAR-beta in cell lines SiHa, HeLa, Caski and C33A were 0.25 +/- 0.08, 0, 0.60 +/- 0.19, 3.12 +/- 0.92 and 0.23 +/- 0.07, 0, 0.14 +/- 0.05, 0.68 +/- 0.21, respectively. The mRNA and protein expression of RAR-beta in SiHa, HeLa and Caski cell lines were decreased or silenced, whereas its expression increased in C33A cell line. MSP method showed that there were RAR-beta gene methylation in SiHa, HeLa and Caski cell lines, while there was no RAR-beta gene methylation in C33A cell line. After treated with 5-Aza-cdR, the mRNA and protein expression levels of RAR-beta in SiHa, HeLa, Caski and C33A cell lines were 1.82 +/- 0.59, 2.13 +/- 0.62, 1.67 +/- 0.43, 2.95 +/- 0.89 and 0.69 +/- 0.21, 0.83 +/- 0.29, 0.56 +/- 0.16, 0.64 +/- 0.20 respectively. The mRNA and protein levels of RAR-beta had a significant difference between before and after interference with 5-Aza-cdR in SiHa, HeLa, and Caski cell lines (P < 0.05). However, they had no significant difference between before and after interference with 5-Aza-cdR in C33A cell line (P > 0.05). The 5-Aza-cdR treatment could suppress cell proliferation. CONCLUSIONS: The RAR-beta gene expression defects play an important role in the carcinogenesis of cervical cancer. Aberrant methylation in promotor region of RAR-beta gene may be an important mechanism for the loss of expression of RAR-beta gene.

Triterpenoids from Coluria longifolia.

A new triterpenoid named 19-hydroxy swinhoeic acid, together with four known compounds were isolated from the whole plants of Coluria longifolia Maxim. Their structures were elucidated on the basis of spectroscopic data analysis and comparison with previously reported data. Cytotoxicities of the five compounds against HepG2 cell lines were also evaluated, unfortunately, no obvious activities were observed.

The crystal structure and photocatalytic properties of a three-dimensional cadmium(II) metal-organic framework: poly[bis(µ3-benzene-1,2-dicarboxylato)[µ2-1,4-bis(pyridin-3-ylmethoxy)benzene]dicadmium(II)].

Photocatalysis is a green technology for the treatment of all kinds of contaminants and has advantages over other treatment methods. Recently, much effort has been devoted to developing new photocatalytic materials based on metal-organic frameworks for use in the degradation of many kinds of organic contaminants. With the aim of searching for more effective photocatalysts, the title three-dimensional coordination polymer, [Cd2(C8H4O4)2(C18H16N2O2)]n, was prepared. The asymmetric unit contains one Cd(II) cation, one benzene-1,2-dicarboxylate anion (denoted L(2-)) and half of a centrosymmetric 1,4-bis(pyridin-3-ylmethoxy)benzene ligand (denoted bpmb). Each Cd(II) centre is five-coordinated by four carboxylate O atoms from two L(2-) ligands and by one N atom from a bpmb ligand, forming a disordered pentagonal pyramidal coordination geometry. The Cd(II) centres are interlinked by L(2-) ligands to form a one-dimensional [Cd2L2]n chain. Adjacent chains are further connected by bpmb linkers, giving rise to a two-dimensional network, and these networks are pillared by bpmb to afford a three-dimensional framework with a 3(3).4(2).6(3).7(1).8(1) topology. Each grid in the framework has large channels which are filled mainly by the two other equivalent frameworks to form a threefold interpenetrating net. The compound exhibits relatively good photocatalytic activity towards the degradation of methylene blue in aqueous solution under UV irradiation.

Studies on the interaction between promethazine and human serum albumin in the presence of flavonoids by spectroscopic and molecular modeling techniques.

Fluorescence, absorption, time-correlated single photon counting (TCSPC), and circular dichroism (CD) spectroscopic techniques as well as molecular modeling methods were used to study the binding characterization of promethazine (PMT) to human serum albumin (HSA) and the influence of flavonoids, rutin and baicalin, on their affinity. The results indicated that the fluorescence quenching mechanism of HSA by PMT is a static quenching due to the formation of complex. The reaction was spontaneous and mainly mediated by hydrogen bonds and hydrophobic interactions. The binding distance between the tryptophan residue of HSA and PMT is less than 8nm, which indicated that the energy transfer from the tryptophan residue of HSA to PMT occurred. The binding site of PMT on HSA was located in sites I and the presence of PMT can cause the conformational changes of HSA. There was the competitive binding to HSA between PMT and flavonoids because of the overlap of binding sites in HSA. The flavonoids could decrease the association constant and increase the binding distance. In addition, their synergistic effect can further change the conformation of HSA. The decrease in the affinities of PMT binding to HSA in the presence of flavonoids may lead to the increase of free drug in blood, which would affect the transportation or disposition of drug and evoke an adverse or toxic effect. Hence, rationalising dosage and diet regimens should be taken into account in clinical application of PMT.

Sonochemical degradation of methyl orange in the presence of Bi2WO6: Effect of operating parameters and the generated reactive oxygen species.

The Bi2WO6 was prepared by the hydrothermal method and its sonocatalytic activity was studied in the degradation of methyl orange (MO) solutions. The effects of catalytic activity of Bi2WO6 on dye were inspected by the change in absorbance of dye with UV-vis spectrometer. The influences of operational parameters such as the addition amount of Bi2WO6, pH, the initial concentration of dyes, ultrasonic power and irradiation time on the degradation ratio were investigated. In addition, the obtained results indicated that the kinetics of sonochemical reactions of MO were consistent with the pseudo first-order kinetics and Bi2WO6 had excellent reusability and stability during the sonochemical degradation processes. The generation and kinds of reactive oxygen species (ROS) and their influence on the sonochemical degradation of MO were determined by the methods of oxidation-extraction spectrophotometry and ROS scavengers. The results indicate that the degradation of MO in the presence of Bi2WO6 under ultrasonic irradiation is related to the generation of ROS, in which both singlet molecular oxygen ((1)O2) and hydroxyl radical (OH) play important roles in the sonochemical degradation of MO. These experimental results provide a sound foundation for the further development of Bi2WO6 as a sonocatalyst in wastewater treatment.