Microbial-Driven Synthesis and Hydrolysis of Neohesperidin Dihydrochalcone: Biotransformation Process and Feasibility Investigation.

A novel yeast-mediated hydrogenation was developed for the synthesis of neohesperidin dihydrochalcone (NHDC) in high yields (over 83%). Moreover, whole-cell catalytic hydrolysis was also designed to hydrolyze NHDC into potential sweeteners, hesperetin dihydrochalcone-7-O-glucoside (HDC-G) and hesperetin dihydrochalcone (HDC). The biohydrogenation was further combined with whole-cell hydrolysis to achieve a one-pot two-step biosynthesis, utilizing yeast to hydrogenate C═C in the structure, while Aspergillus niger cells hydrolyze glycosides. The conversion of NHDC and the proportion of hydrolysis products could be controlled by adjusting the catalysts, the components of the reaction system, and the addition of glucose. Furthermore, yeast-mediated biotransformation demonstrated superior reaction stability and enhanced safety and employed more cost-effective catalysts compared to the traditional chemical hydrogenation of NHDC synthesis. This research not only provides a new route for NHDC production but also offers a safe and flexible one-pot cascade biosynthetic platform for the production of high-value compounds from citrus processing wastes.

Spectrally efficient multi-service fiber-wireless access in low-cost direct-detection THz system at 300†GHz.

This Letter demonstrates a novel, to the best of knowledge, overlapping single-sideband (OSSB) transmission scheme for spectrally efficient multi-service fiber-wireless (FiWi) access in a low-cost direct-detection (DD) THz system. Utilizing the proposed OSSB scheme, user data from different services can share the same spectrum resource yet can be successfully demodulated via one cost-effective DD THz receiver in conjunction with the Kramers-Kronig (KK) based SSB field reconstruction and look-up table (LUT) enabled signal separation algorithms. A proof-of-principle experiment is conducted. Based on an IQ modulator and a single THz zero-bias diode (ZBD), two independent 10-GBd quadrature phase shift keying (QPSK) signals with an overlapped spectrum are successfully demodulated after 20-km fiber and up to 3-m wireless transmission at the 300-GHz band. To the best of our knowledge, this is the first demonstration of multi-service FiWi access with an OSSB format in a 300-GHz DD THz system.

DNA methylation of DKK-1 may correlate with pathological bone formation in ankylosing spondylitis.

OBJECTIVE: To investigate DNA methylation (DNAm) status of dickkopf-associated protein 1 (DKK-1) in ossified hip capsule synovium and serum among patients with ankylosing spondylitis (AS). METHODS: Western blot was applied to detect the level of DKK-1 protein expression in hip joint capsule tissues from four patients with AS as well as four patients with femoral neck fracture (FNF) caused by trauma as control. DKK-1 gene promoter methylation (GPM) was examined by methylation-specific polymerase chain reaction. Reverse transcription-polymerase chain reaction was performed to examine the messenger RNA (mRNA) levels of DKK-1, ß-catenin, and Wnt3a in both tissue and serum. The DNAm status of serum DKK-1 was measured among 36 patients with AS and syndesmophytes (AS + syndesmophytes group), 40 patients with AS but no syndesmophyte (AS group), and 42 healthy individuals (control group). Also, the serum levels of DKK-1 were measured by enzyme-linked immunosorbent assay. The modified New York criteria (mNYC) together with the modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS) were adopted to examine the radiographic progression of AS. The receiver operating characteristic (ROC) curve was applied to investigate the diagnostic value of the methylation rate of DKK-1 with regard to radiographic progression. RESULTS: The expressions of DKK-1 protein and mRNA in hip joint capsule tissues of AS patients were significantly lower, while DKK-1 GPM rate, ß-catenin mRNA, and Wnt3a mRNA were markedly higher when compared with FNF group. For serum samples, the DKK-1 methylation rate was significantly higher in AS+ syndesmophytes group in contrast to AS group and healthy controls. Serum levels of DKK-1 protein and mRNA in AS with syndesmophytes group were markedly decreased, while ß-catenin mRNA and Wnt3a mRNA expressions were significantly increased than AS with no syndesmophyte group and the healthy control group. AS patients in Grade 4 showed a significantly higher serum DKK-1 GPM rate than those in Grade 3 based on mNYC. Serum DKK-1 GPM level was markedly and positively correlated with mSASSS. Serum levels of DKK-1 in AS+ syndesmophytes group were markedly lower compared with AS but no syndesmophyte group and healthy controls. ROC curve analysis indicated that serum DKK-1 methylation rate serves as a decent indicator for AS radiographic progression. CONCLUSION: DNAm of DKK-1 may correlate with pathological bone formation in AS, which may provide new strategies for the treatment of AS abnormal bone formation.

Nonlinearity mitigation in a fiber-wireless integrated system based on low-complexity autoencoder and BiLSTM-ANN equalizer.

We propose and experimentally demonstrate an intelligent nonlinear compensation method using a stacked autoencoder (SAE) model in conjunction with principal component analysis (PCA) technology and a bidirectional long-short-term memory coupled with ANN (BiLSTM-ANN) nonlinear equalizer for an end-to-end (E2E) fiber-wireless integrated system. The SAE-optimized nonlinear constellation is utilized to mitigate nonlinearity during the optical and electrical conversion process. Our proposed BiLSTM-ANN equalizer is primarily based on time memory and information extraction characteristics, which can compensate for the remaining nonlinear redundancy. A low-complexity 50 Gbps E2E-optimized nonlinear 32 QAM signal is successfully transmitted over a span of 20 km standard single-mode fiber (SSMF) and 6 m wireless link at 92.5 GHz. The extended experimental results indicate that the proposed E2E system can achieve a reduction of up to 78% in BER and a gain in receiver sensitivity of over 0.7 dB at BER of 3.8 × 10-3. Moreover, computational complexity is reduced by more than 10 times compared to the classical training model.

Analysis of circular RNA expression profile of pathological bone formation in ankylosing spondylitis.

OBJECTIVE: To screen and analyze the function of specific CircRNAs involved in pathological bone formation in patients with ankylosing spondylitis (AS). METHODS: From September 2019 to October 2020, hip capsule tissues obtained from 3 patients with AS developed hip joint fusion and 3 patients with femoral neck fracture (FNF) were obtained. The circular RNA expressions of hip capsule were analyzed by Arraystar CircRNA chip. qRT-PCR analysis wan performed to identify the expression patterns of differentially expression CircRNAs. RESULTS: Our findings showed that there were 25 up-regulated and 39 down-regulated differential CircRNAs. Among these CircRNAs, we screened 10 highest up-regulatedCircRNAs and 13 lowest down-regulated CircRNAs (Fold Change≥2,P<0.05). In further verification analysis, hsa_circ_0067103, hsa_circ_0004496, and hsa_circ_0002649, ACTG1 were significantly upregulated, while hsa_circ_0020273, hsa_circ_0005699, and hsa_circ_0048764 were markly downregulated in AS tissue than FNF controls. CONCLUSION: The expression of CircRNAs involved of pathological bone formation in AS were significantly different from those of control group. These differentially expressed Circular RNAs may be closely related to the occurrence and development of pathological bone formation in AS.

Real-time 100-GbE fiber-wireless seamless integration system using an electromagnetic dual-polarized single-input single-output wireless link at the W band.

This Letter demonstrates a real-time 100-GbE fiber-wireless seamless integration system operating at the whole W band (75-110 GHz). Based on a pair of commercial digital coherent optical modules, the real-time transparent transmission of 125-Gb/s dual-polarized quadrature phase-shift keying signal has been successfully achieved over two-spans of 20-km fiber and up to 150-m electromagnetic dual-polarized single-input single-output wireless link. To the best of our knowledge, this is the first real-time demonstration of 100-GbE signal transmission over >100-m wireless distance at the millimeter-wave band based on photonics. We believed this real-time and high-speed fiber-wireless seamless integration system with a wireless coverage up to hundreds of meters can significantly accelerate the progress of upcoming 6G.

Photonics-aided integrated sensing and communications in mmW bands based on a DC-offset QPSK-encoded LFMCW.

The evolution of mobile communications towards millimeter-wave (mmW) bands provides a strong opportunity for the seamless integration of radar and wireless communications. We present a photonics-aided mmW integrated sensing and communications (ISAC) system constructed by photonic up-conversion using a coherent optical frequency comb, which facilitates zero frequency offset of the resulting mmW signal. The sensing and communications functions are enabled by a joint waveform that encodes a DC-offset QPSK signal on a linear frequency-modulated continuous wave (LFMCW) in baseband. The QPSK encoding ensures the constant envelope of the mmW ISAC signal for long-distance radar detection. The optimized DC offset preserves the distinctive chirp phase and good cross-correlation of the original LFMCW, which can achieve high-resolution sensing by radar de-chirping and assist in communication sequence synchronization by pulse compression, respectively. Experimental results show that the single-user detection with less than 20-mm sensing error and dual-user detection with a 10.4-cm ranging resolution are realized at 28-GHz band, respectively. The wireless communication with a 11.5-Gbit/s transmission rate also at 28-GHz band is successfully tested. Moreover, the proof-of-concept experiments demonstrate the good frequency tunability and wavelength tolerance of the proposed ISAC system.

Demonstration of 144-Gbps Photonics-Assisted THz Wireless Transmission at 500 GHz Enabled by Joint DBN Equalizer.

The THz wireless transmission system based on photonics has been a promising candidate for further 6G communication, which can provide hundreds of Gbps or even Tbps data capacity. In this paper, 144-Gbps dual polarization quadrature-phase-shift-keying (DP-QPSK) signal generation and transmission over a 20-km SSMF and 3-m wireless 2 × 2 multiple-input multiple-output (MIMO) link at 500 GHz have been demonstrated. To further compensate for the linear and nonlinear distortions during the fiber-wireless transmission, a novel joint Deep Belief Network (J-DBN) equalizer is proposed. Our proposed J-DBN-based schemes are mainly optimized based upon the constant modulus algorithm (CMA) and direct-detection least mean square (DD-LMS) equalization. The results indicate that the J-DBN equalizer has better bit error rate (BER) performance in receiver sensitivity. In addition, the computational complexity of the J-DBN-based equalizer can be approximately 46% lower than that of conventional equalizers with similar performance. To our knowledge, this is the first time that a novel joint DBN equalizer has been proposed based on classical algorithms. It is a promising scheme to meet the demands of future fiber-wireless integration communication for low power consumption, low cost, and high capacity.

Comparative transcriptomics to reveal the mechanism of enhanced catalytic activities of Aspergillus niger whole-cells cultured with different inducers in hydrolysis of citrus flavonoids.

A new Aspergillus niger whole-cell catalyst was cultured for the cascade hydrolysis of hesperidin (HES) to produce high-value hesperetin-7-O-glucoside (HG) and hesperetin with high conversion (above 90%). Moreover, the inducers used were shown to be useful for cell growth and to induce cells to produce specific enzymes. Remarkably, the type of inducers determined whether the cells can hydrolyze HES. The product composition was also controllable by adjusting different inducers. Transcriptome analysis suggested that both naringin-vs-blank group and saccharose-vs-blank group had obviously difference in gene expression. The naringin-vs-blank group was mainly up-regulated differentially expressed genes (DEGs), while saccharose-vs-blank group was mainly down-regulated DEGs. The Gene Ontology (GO) analysis showed that whether naringin or saccharose was added as an inducer would greatly affect the catalytic activity of cells. Furthermore, 3 genes related to rhamnosidase, 14 genes related to glucosidase and 5 genes related to hydrolase activity were found. These genes were not only involved in rhamnosidase and glucosidase activities, but also spliceosome and the sucrose and starch metabolic pathways. The quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the results of transcriptome sequencing were reliable. This study gave a new approach to hydrolyze HES, and new perspectives to understand the mechanisms associated with the hydrolysis of whole-cell catalyst.

Engineering Multifunctional Hydrogel With Osteogenic Capacity for Critical-Size Segmental Bone Defect Repair.

Treating critical-size segmental bone defects is an arduous challenge in clinical work. Preparation of bone graft substitutes with notable osteoinductive properties is a feasible strategy for critical-size bone defects. Herein, a biocompatible hydrogel was designed by dynamic supramolecular assembly of polyvinyl alcohol (PVA), sodium tetraborate (Na2B4O7), and tetraethyl orthosilicate (TEOS). The characteristics of the supramolecular hydrogel were evaluated by rheological analysis, swelling ratio, degradation experiments, and scanning electron microscopy (SEM). In in vitro experiments, this TEOS-hydrogel had self-healing property, low swelling rate, degradability, good biocompatibility, and induced osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by upregulating the expression of Runx-2, Col-1, OCN, and osteopontin (OPN). In segmental bone defect rabbit models, the TEOS-containing hydrogel accelerated bone regeneration, thus restoring the continuity of bone and recanalization of the medullary cavity. The abovementioned results demonstrated that this TEOS-hydrogel has the potential to realize bone healing in critical-size segmental bone defects.

Tyrosinase inhibitory mechanism and the anti-browning properties of piceid and its ester.

Different mechanisms for inhibiting tyrosinase can be exploited to avoid quality losses caused by the enzymatic browning of fruits and vegetables. Piceid (PI) and piceid 6″-O- azelaic acid ester (PIA) are oxidized by tyrosinase; however, their oxidation products may have inhibitory effects on tyrosinase. This notion is because l-DOPA oxidation was inhibited after the pre-incubation of PI/PIA with tyrosinase, however, l-DOPA oxidation was not affected if this pre-incubation was not performed. Circular dichroism analysis indicated a conformational change in the secondary structure of tyrosinase after pre-incubation. Further, molecular docking and enzyme reaction kinetics assays were employed to reveal the mechanism underlying the effects of PI/PIA on tyrosinase in the absence of pre-incubation with tyrosinase. PI/PIA had anti-browning effects in the potato models. The increased rate of A420 in PI/PIA groups at 24 h were 281% and 279%, which were approximately 2.4- and 2.5-fold lower than that of control (668%).

Real-time demonstration of 103.125-Gbps fiber-THz-fiber 2 × 2 MIMO transparent transmission at 360-430 GHz based on photonics.

In this Letter, we experimentally demonstrate the first real-time transparent fiber-THz-fiber 2 × 2 multiple-input multiple-output (MIMO) transmission system with a record line rate of 125.516 Gbps at 360-430 GHz based on photonic remote heterodyning, hybrid optoelectronic down-conversion, and commercial digital coherent modules. The 103.125-Gbps net data rate using dual-polarization quadrature phase-shift keying (DP-QPSK) modulation is successfully transmitted over two spans of 20-km standard single-mode fiber (SSMF) and 60-cm wireless distance under 15% soft-decision forward error correction (SD-FEC) for a pre-FEC bit error ratio (BER) threshold of 1.56 × 10-2 (post-FEC BER < 10-15). The optical signal to noise ratio (OSNR) margin and the stability of the transmission system are extensively investigated. To the best of our knowledge, this is the first time to realize >100-Gbps real-time transparent fiber-THz-fiber link transmission at beyond the 350-GHz band, making it a promising scheme to pave the way towards a practical seamless integration of a fiber-THz-fiber link to the future 6G mobile communication system.

Decreased serum CXCL12/SDF-1 concentrations may reflect disease severity of non-traumatic osteonecrosis of femoral head.

OBJECTIVE: The current study was performed to investigate the potential association of serum CXCL12 with disease severity in non-traumatic ONFH. METHODS: This study enrolled 182 patients with non-traumatic ONFH and 182 age- and gender-matched healthy controls. The CXCL12 levels in serum were measured by enzyme-linked immunosorbent assay. Meanwhile, serum levels of procollagen type I (PINP) and Interleukin-33(IL-33) were also detected. The radiographic severity was determined by FICAT grade. Clinical severity was evaluated by visual analogue scale (VAS), Harris Hip Score (HHS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Among the non-traumatic ONFH, 90 patients ONFH received total hip arthroplasty, the localization and expression of the CXCL12 protein and mRNA were detected by immunohistochemistry, Western blot analysis, RT-PCR and in necrotic area and adjacent non-necrotic area from lesioned femoral neck from ONFH patients and healthy femoral head from femoral neck fracture patients. Receiver operating characteristic (ROC) curve analysis was carried out to confirm the diagnostic value serum CXCL12, PINP and IL-33 with regard to the FICAT grade. RESULTS: Serum CXCL12 levels were significantly lower in non-traumatic ONFH patients compared with healthy controls. CXCL12 mRNA and protein expressions were both significantly decreased in necrotic area in comparison with non-necrotic area and healthy femoral head. Serum CXCL12 concentrations were drastically reduced in patients with FICAT stage 4 compared with stage 3, and CXCL12 concentrations in patients with stage 3 were markedly lower than stage 2. Serum CXCL12 levels were negatively related to FICAT grading. In addition, Serum CXCL12 concentrations were also negatively related to VAS, WOMAC scores and positively correlated with HHS scores. Meanwhile, serum CXCL12 levels were positively correlated with serum PINP and negatively correlated with IL-33 levels. ROC curve analysis implicated that decrease CXCL12 in serum may act as a favorable marker for FICAT grade. CONCLUSIONS: Decreased serum CXCL12 concentrations may reflect disease severity of non-traumatic ONFH.

In vitro absorption and lipid-lowering activity of baicalin esters synthesized by whole-cell catalyzed esterification.

Baicalin, a phenolic glycoside with good lipid-lowering activity, has poor intestinal absorption due to low lipophilicity. In this study, six ester derivatives of baicalin, named BECn (n = 2, 3, 4, 6, 8, and 10) based on their fatty chain lengths, were synthesized by whole-cell catalyzed esterification to improve lipophilicity, and the intestinal absorption and lipid-lowering activity of the synthesized esters were investigated using cell models in vitro. BEC2, BEC3, and BEC4 exhibited higher Papp values than baicalin in Caco-2 cell monolayers. The lipid-lowering activity of the three esters was stronger than baicalin in the cell models of hepatic steatosis, adipocytes and foam macrophages, and was attributed to their higher intracellular accumulation and stronger direct activation of the carnitine palmitoyltransferase 1A. Moreover, these esters were easily hydrolyzed by carboxylesterase and were unstable at pH 7.4, which significantly weakened their absorption and lipid-lowering activity. This study laid the foundation for industrial production and practical application of BAI esters.

Increased serum CXCL10 levels are associated with clinical severity and radiographic progression in patients with lumbar disc degeneration.

BACKGROUND: Lumbar intervertebral degenerative disc disease (IDD) is a multifaceted progressive condition that commonly occurs in conjunction with lumbar disc herniation (LDH). CXCL10 mRNA appears to be increased in both IDD and LHD. OBJECTIVE: This study was performed to identify the relationship between serum CXCL10 levels and disease severity in patients with IDD. METHODS: 136 IDD patients with low back pain, 127 asymptomatic volunteers and 120 healthy controls were enrolled. Serum CXCL10 protein concentrations were detected using commercial human CXCL10 ELISA Kits. Serum CXCL10 mRNA were examined using qRT-PCR. Clinical severity was assessed using the visual analog scale (VAS) and Oswestry Disability Index(ODI) scores. Radiographic severity was defined using the MRI-based Pfirrmann classification of disc degeneration. Receiver operating characteristic (ROC) curve analysis was used in estimating the correlation between CXCL10 and Pfirrmann grade. The cross-sectional area (CSA) of the lumbar multifidus muscle (LMM) and psoas major (PM) were calculated, and fat infiltration was evaluated by Ropponen-Kjaer criteria. RESULTS: Serum CXCL10 concentrations were markedly raised in IDD patients with low back pain in contrast to asymptomatic individuals and healthy controls. Serum CXCL10 levels were positively associated with Pfirrmann grade. ROC curve analysis indicated that serum CXCL10 correlated well with Pfirrmann grade. In addition, serum CXCL10 concentrations were significantly higher in IDD patients with LMM and PM degeneration compared with IDD patients without degeneration. Increased CXCL10 levels positively correlated with VAS and ODI scores, as well as decreased CSA and fat filtration of the LMM and PM. CONCLUSION: Increased serum CXCL10 levels correspond to clinical severity and radiographic progression in IDD patients.

Efficient Enzymatic Synthesis of Lipophilic Phenolic Glycoside Azelaic Acid Esters and Their Depigmenting Activity.

In this paper, an enzymatic route for synthesizing phenolic glycoside azelaic acid esters was successfully set up via lipase-catalyzed esterification and transesterification. Among the lipases tested, Candida antarctica lipase B (Novozyme 435) showed the highest activity in catalyzing esterification and Thermomyces lanuginosus (Lipozyme TLIM) gave the highest substrate conversion in catalyzing transesterification for the synthesis of ester. The addition of 4A molecular sieves into the reaction system is found to be an effective method for in situ absorption of the byproduct water and methanol, with which the substrate conversions of the enzymatic esterification and transesterification were 98.7 and 95.1%, respectively. Also, the main product ratios in transesterification were above 99.0% with lipozyme TLIM as a catalyst because the hydrolysis reaction was hindered. The results of the physical and biological properties indicate that all esters had higher Clog p values than their parent compounds. Also, the esters showed higher intracellular tyrosinase inhibitory and depigmentating activities than phenolic glycosides, azelaic acid (AA), and their physical mixtures due to their higher membrane penetration and tyrosinase inhibitory effects. In particular, piceid 6″-O-azelaic acid ester (PIA) showed the strongest inhibitory effect against melanin production. Its inhibitory rate was 77.4% at a concentration of 0.25 mM, about 4.2 times higher than that of arbutin (18.5%).

Tail suspension delays ectopic ossification in proteoglycan-induced ankylosing spondylitis in mice via miR-103/DKK1.

Ankylosing spondylitis (AS), characterized by inflammatory lesions and osteophyte formation, is a common immune rheumatic disease affecting the sacroiliac and axial joints. A high-intensity mechanical load is known to accelerate the heterotopic ossification associated with enthesitis in AS. Thus, the present study explored whether decreased mechanical load could delay the heterotopic ossification in AS. First, 24-week-old female BALB/c mice were induced with proteoglycan (PG) to establish an AS model. The AS-induced pathological and bone morphological changes of the sacroiliac joint were confirmed by hematoxylin and eosin staining and microCT analysis, respectively. Subsequently, the mice were treated with interventions of different mechanical loads. Using reverse transcription-quantitative PCR, it was revealed that expression levels of the osteogenesis-related genes bone morphogenetic protein-2, runt-related transcription factor 2 and osteocalcin were significantly reduced in sacroiliac bone tissue after intervention with a reduced mechanical load. The level of mechanosensory microRNA (miR)-103 increased in response to reduced mechanical loads. Consistently, in groups with reduced mechanical load, proteins with mechanical functions, including ρ-associated coiled-coil-containing protein kinase 1 (ROCK1), phosphorylated (p)-Erk1/2 and ß-catenin, were reduced compared with the PG control. A dual-luciferase assay verified that miR-103 binds to the 3'-untranslated region end of Rock1 mRNA, thus negatively regulating the activity of Rock1 and affecting pathological ossification during AS. However, immunohistochemical staining indicated that the expression of dickkopf Wnt signaling pathway inhibitor 1, an inhibitor of the Wnt/ß-catenin pathway, was increased in sacroiliac tissues. The results indicated that tail suspension decreased the mechanical load, thus reducing the bone formation in AS mice. Furthermore, tail suspension could inhibit the activation of mechanical kinase ROCK1 and p-Erk1/2 in the MAPK signaling pathway by upregulating miR-103, thereby inhibiting the classical osteogenesis-related Wnt/ß-catenin pathway in AS. In summary, the present study uncovered the ameliorative effect of suspension on AS and its therapeutic potential for AS.

Antityrosinase Mechanism and Antimelanogenic Effect of Arbutin Esters Synthesis Catalyzed by Whole-Cell Biocatalyst.

Tyrosinase is a key enzyme responsible for enzymatic browning of fruits and vegetables and skin disorders due to overproduction of melanin. Arbutin is an inhibitor of tyrosinase; however, its high polarity and weak transdermal absorption capacity limit its applications. In this paper, a green solvent system was developed to successfully synthesize arbutin esters with improved liposolubilities (Clog P values = 0.27-5.03). Among the obtained esters, arbutin undecenoate (AU) showed the strongest tyrosinase-inhibiting activity (15.6%), which was 9.0 times higher than that of arbutin. An enzyme kinetics study indicated that AU was a competitive inhibitor with reversible inhibition. The esters inhibited tyrosinase by making the secondary structure of tyrosinase looser and less stable; moreover, the interactions between tyrosinase and AU driven by metal interactions and hydrogen bonds also offered a mechanism for inhibition of AU on tyrosinase. In addition, AU (100 µM) reduced the melanin content of B16 mouse melanoma cells to 61.3% of the control group.

Endocannabinoid signaling in the lateral habenula regulates pain and alcohol consumption.

Hyperalgesia, which often occurs in people suffering from alcohol use disorder, may drive excessive drinking and relapse. Emerging evidence suggests that the lateral habenula (LHb) may play a significant role in this condition. Previous research suggests that endocannabinoid signaling (eCBs) is involved in drug addiction and pain, and that the LHb contains core components of the eCBs machinery. We report here our findings in rats subjected to chronic ethanol vapor exposure. We detected a substantial increase in endocannabinoid-related genes, including Mgll and Daglb mRNA levels, as well as monoacylglycerol lipase (MAGL) protein levels, as well as a decrease in Cnr1 mRNA and type-1 cannabinoid receptor (CB1R) protein levels, in the LHb of ethanol-exposed rats. Also, rats withdrawing from ethanol exposure displayed hypersensitivity to mechanical and thermal nociceptive stimuli. Conversely, intra-LHb injection of the MAGL inhibitor JZL184, the fatty acid amide hydrolase inhibitor URB597, or the CB1R agonist WIN55,212-2 produced an analgesic effect, regardless of ethanol or air exposure history, implying that alcohol exposure does not change eCB pain responses. Intra-LHb infusion of the CB1R inverse agonist rimonabant eliminated the analgesic effect of these chemicals. Rimonabant alone elicited hyperalgesia in the air-, but not ethanol-exposed animals. Moreover, intra-LHb JZL184, URB597, or WIN55,212-2 reduced ethanol consumption in both homecages and operant chambers in rats exposed to ethanol vapor but not air. These findings suggest that LHb eCBs play a pivotal role in nociception and facilitating LHb eCBs may attenuate pain in drinkers.

CircCDR1as Suppresses Bone Microvascular Endothelial Cell Activity and Angiogenesis Through Targeting miR-135b/ FIH-1 Axis.

OBJECTIVE: The current study investigated the role of CircCDR1as on angiogenesis of bone microvascular endothelial cells (BMECs) isolated from non-traumatic ONFH. METHODS: Forty corticosteroid-induced ONFH patients received THA were enrolled in our study. Expressions of CircCDR1as, miR-135b, and FIH-1 were detected by qRT-PCR in affected necrosis tissue and non-affected normal tissue. Bone microvascular endothelial cells (BMEC) were isolated from six patients and treated with 0.1 mg/mL hydrocortisone to establish a GC-damaged model of BMECs. Circ CDR1as plasmid and miR-135b mimic were transfected into BMECs. BMEC proliferation was assessed using MTT assays. The migration ability of cells was detected by scratch-wound assays. Matrigel assay was performed to detect angiogenesis in vitro. Western blot assay was used to detect HIF-1α, VEGF, and FIH-1 expressions. FISH, RNA pull down, RIP, and luciferase assay were carried out to determine the interaction of CircCDR1as, miR-135b, and FIH-1. RESULTS: CircCDR1as was upregulated(2.02 ± 0.30 vs. 1.00 ± 0.10,P < 0.001) whereas miR-135b was downregulated (0.55 ± 0.12 vs. 1.00 ± 0.10,P < 0.001) in affected tissues than in non-affected tissues. Expression of CircCDR1as and FIH-1 were negatively associated with miR-135b in affected tissues (CircCDR1as with miR-135b: r = -0.506, P < 0.001; FIH-1 with miR-135b r = -0.510, P < 0.001). Total blood tubule density was increased when CircCDR1as was silenced compared with NC (P < 0.01 vs. NC). The number of migrated BMECs were significantly increased in CircCDR1as silencing group compared with NC group (P < 0.05 vs. NC). In addition, CircCDR1as plasmids transfection increased the protein expressions of FIH-1 (P < 0.05 vs. NC) and reduced the HIF-1α as well as VEGF expression compared with NC group (P < 0.05 vs. NC). FISH, RNA pull down, RIP, and luciferase assay identified that FIH-1 was a target of miR-135b and could be modulated by CircCDR1as. CONCLUSION: CircCDR1as decreases angiogenesis and proliferation of BMECs by sponging miR-135b and upregulate FIH-1.