Discovery and characterization of a specific inhibitor of serine-threonine kinase cyclin-dependent kinase-like 5 (CDKL5) demonstrates role in hippocampal CA1 physiology.

Pathological loss-of-function mutations in cyclin-dependent kinase-like 5 (CDKL5) cause CDKL5 deficiency disorder (CDD), a rare and severe neurodevelopmental disorder associated with severe and medically refractory early-life epilepsy, motor, cognitive, visual, and autonomic disturbances in the absence of any structural brain pathology. Analysis of genetic variants in CDD has indicated that CDKL5 kinase function is central to disease pathology. CDKL5 encodes a serine-threonine kinase with significant homology to GSK3ß, which has also been linked to synaptic function. Further, Cdkl5 knock-out rodents have increased GSK3ß activity and often increased long-term potentiation (LTP). Thus, development of a specific CDKL5 inhibitor must be careful to exclude cross-talk with GSK3ß activity. We synthesized and characterized specific, high-affinity inhibitors of CDKL5 that do not have detectable activity for GSK3ß. These compounds are very soluble in water but blood-brain barrier penetration is low. In rat hippocampal brain slices, acute inhibition of CDKL5 selectively reduces postsynaptic function of AMPA-type glutamate receptors in a dose-dependent manner. Acute inhibition of CDKL5 reduces hippocampal LTP. These studies provide new tools and insights into the role of CDKL5 as a newly appreciated key kinase necessary for synaptic plasticity. Comparisons to rodent knock-out studies suggest that compensatory changes have limited the understanding of the roles of CDKL5 in synaptic physiology, plasticity, and human neuropathology.

Discovery and characterization of a specific inhibitor of serine-threonine kinase cyclin dependent kinase-like 5 (CDKL5) demonstrates role in hippocampal CA1 physiology.

Pathological loss-of-function mutations in cyclin-dependent kinase-like 5 ( CDKL5 ) cause CDKL5 deficiency disorder (CDD), a rare and severe neurodevelopmental disorder associated with severe and medically refractory early-life epilepsy, motor, cognitive, visual and autonomic disturbances in the absence of any structural brain pathology. Analysis of genetic variants in CDD have indicated that CDKL5 kinase function is central to disease pathology. CDKL5 encodes a serine-threonine kinase with significant homology to GSK3b, which has also been linked to synaptic function. Further, Cdkl5 knock-out rodents have increased GSK3b activity and often increased long-term potentiation (LTP). Thus, development of a specific CDKL5 inhibitor must be careful to exclude cross-talk with GSK3b activity. We synthesized and characterized specific, high-affinity inhibitors of CDKL5 that do not have detectable activity for GSK3b. These compounds are very soluble in water but blood-brain barrier penetration is low. In rat hippocampal brain slices, acute inhibition of CDKL5 selectively reduces post-synaptic function of AMPA-type glutamate receptors in a dose-dependent manner. Acute inhibition of CDKL5 reduces hippocampal LTP. These studies provide new tools and insights into the role of CDKL5 as a newly appreciated, key kinase necessary for synaptic plasticity. Comparisons to rodent knock-out studies suggest that compensatory changes have limited the understanding of the roles of CDKL5 in synaptic physiology, plasticity and human neuropathology.

Effective method for low-light image enhancement based on the JND and OCTM models.

Low-light images always suffer from dim overall brightness, low contrast, and low dynamic ranges, thus result in image degradation. In this paper, we propose an effective method for low-light image enhancement based on the just-noticeable-difference (JND) and the optimal contrast-tone mapping (OCTM) models. First, the guided filter decomposes the original images into base and detail images. After this filtering, detail images are processed based on the visual masking model to enhance details effectively. At the same time, the brightness of base images is adjusted based on the JND and OCTM models. Finally, we propose a new method to generate a sequence of artificial images to adjust the brightness of the output, which has a better performance in image detail preservation compared with other single-input algorithms. Experiments have demonstrated that the proposed method not only achieves low-light image enhancement, but also outperforms state-of-the-art methods qualitatively and quantitatively.

Risk Genetic Variants (IL-10) for Osteoporosis in Han Population from Northwest China.

Background: Osteoporosis (OP) is a common metabolic bone disease characterized by loss of bone mass. IL-10 is considered to be a powerful immune and inflammatory suppressor. This study aimed to assess association between genetic loci in IL-10 and susceptibility to OP. Methods: Association analysis between IL-10 genetic loci and OP risk through SNPStats online software. FPRP analysis (false-positive report probability) verified whether the positive results were noteworthy findings. Linkage disequilibrium (LD) and haplotype analysis were completed by Haploview 4.2 and SNPStats. Multi-factor dimensionality reduction (MDR) was used to assess interaction of SNP-SNP in susceptibility to OP. Results: Allele "G" of IL-10-rs1554286 (OR = 1.21, p = 0.013), allele "C" of IL-10-rs1518111 (OR = 1.22, p = 0.011), allele "C" of IL-10-rs3024490 (OR = 1.20, p = 0.018), and allele "G" of IL-10-rs1800871 (OR = 1.21, p = 0.015) were risk factors for OP. In females, smoking, drinking, or aging ≤60 years old participants, the above genetic loci are also significantly associated with the increased risk of OP. FPRP analysis showed that all positive results are noteworthy findings. There are significant differences in serum levels of uric acid, mean hemoglobin concentration, or mean hemoglobin among different genotypes of IL-10 gene loci. MDR showed that four loci model composed rs1554286, rs1518111, rs3021094, and rs1800871 is the best model for predicting OP risk. Conclusion: IL-10-rs1554286, -rs1518111, -rs3021094, and -rs1800871 are risk factors for susceptibility to OP.

Pseudorabies virus exploits N6-methyladenosine modification to promote viral replication.

Introduction: Pseudorabies virus (PRV) is the pathogenic virus of porcine pseudorabies (PR), belonging to the Herpesviridae family. PRV has a wide range of hosts and in recent years has also been reported to infect humans. N6-methyladenosine (m6A) modification is the major pathway of RNA post-transcriptional modification. Whether m6A modification participates in the regulation of PRV replication is unknown. Methods: Here, we investigated that the m6A modification was abundant in the PRV transcripts and PRV infection affected the epitranscriptome of host cells. Knockdown of cellular m6A methyltransferases METTL3 and METTL14 and the specific binding proteins YTHDF2 and YTHDF3 inhibited PRV replication, while silencing of demethylase ALKBH5 promoted PRV output. The overexpression of METTL14 induced more efficient virus proliferation in PRV-infected PK15 cells. Inhibition of m6A modification by 3-deazaadenosine (3-DAA), a m6A modification inhibitor, could significantly reduce viral replication. Results and Discussion: Taken together, m6A modification played a positive role in the regulation of PRV replication and gene expression. Our research revealed m6A modification sites in PRV transcripts and determined that m6A modification dynamically mediated the interaction between PRV and host.

Effective enhancement method of low-light-level images based on the guided filter and multi-scale fusion.

Aiming to solve the problem of low-light-level (LLL) images with dim overall brightness, uneven gray distribution, and low contrast, in this paper, we propose an effective LLL image enhancement method based on the guided filter and multi-scale fusion for contrast enhancement and detail preservation. First, a base image and detail image(s) are obtained by using the guided filter. After this procedure, the base image is processed by a maximum entropy-based Gamma correction to stretch the gray level distribution. Unlike the existing methods, we enhance the detail image(s) based on the guided filter kernel, which reflects the image area information. Finally, a new method is proposed to generate a sequence of artificial images to adjust the brightness of the output, which has a better performance in image detail preservation compared with other single-input algorithms. Experiments show that the proposed method can provide a more significant performance in enhancing contrast, preserving details, and maintaining the natural feeling of the image than the state of the art.

Morinda officinalis polysaccharide attenuates osteoporosis in rats underwent bilateral ovariectomy by suppressing the PGC-1α/PPARγ pathway.

OBJECTIVE: Osteoporosis (OP) is a widespread disease that causes risks of spine and hip fractures. Morinda officinalis polysaccharide (MOP) shows therapeutic potential in OP. This article intended to understand the mechanism by which MOP impacts bone mineral density (BMD) and serum trace elements in OP rats. METHODS: OP rat models were established by bilateral ovariectomy (OVX). Rats were intragastrically administered with MOP or ZLN005 [the activator of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α)] since the first day after operation for 8 weeks. Microstructural changes in OP rats were analyzed using micro-computed tomography system. Contents of serum Zn, Cu, Fe, and Mg in rats were measured. Levels of serum superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), GSH, and malondialdehyde (MDA) in rats were determined by Enzyme-linked immunosorbent assay. Protein levels of PGC-1α and peroxisome proliferator-activated receptor γ (PPARγ) in cartilage tissues of rats were determined via Western blotting. RESULTS: MOP enhanced BMD, bone volume per trabecular volume (BV/TV), Tb.N, and Tb.Th and reduced Tb.Sp in the distal femur of OVX rats, elevated levels of serum Cu, Fe, and Mg and contents of SOD, GSH, and GSH-PX and decreased MDA content. Moreover, MOP suppressed the PGC-1α/PPARγ pathway. Activation of PGC-1α partially abolished the action of MOP on ameliorating OP in OVX rats and strengthening anti-oxidation ability. CONCLUSION: MOP mitigated OP in OVX rats by inhibiting the PGC-1α/PPARγ pathway.

Adaptive method for image dynamic range adjustment and detail enhancement.

Tone mapping operators (TMOs) aim to adjust high dynamic range (HDR) images to low dynamic range (LDR) ones so that they can be displayed on conventional devices with visual information retained. Nonetheless, existing TMOs can successfully tone-map only limited types of HDR images, and the parameters need to be manually adjusted to yield the best subjective-quality tone-mapped outputs. To cope with the aforementioned issues, an adaptive parameter-free and scene-adaptive TMO for dynamic range adjusting and detail enhancing is proposed to yield a high-resolution and high-subjective-quality tone-mapped output. This method is based on detail/base layer decomposition to decompose the input HDR image into coarse detail, fine detail, and base images. After that, we adopt different strategies to process each layer to adjust the overall brightness and contrast and to retain as much scene information. Finally, a new method, to the best of our knowledge, is proposed for visualization to generate a sequence of artificial images to adjust the brightness. Experiments with numerous HDR images and state-of-the-art TMOs are conducted; the results demonstrate that the proposed method consistently produces better quality tone-mapped images than the state-of-the-art methods.

Narcissistic Enough to Challenge: The Effect of Narcissism on Change-Oriented Organizational Citizenship Behavior.

During the COVID-19 pandemic, organizations need to effectively manage changes, and employees need to proactively adapt to these changes. The present research investigated when and how individual employees' narcissism was related to their change-oriented organizational citizenship behavior. Specifically, based on a trait activation perspective, this research proposed the hypotheses that individual employees' narcissism and environmental uncertainty would interactively influence employees' change-oriented organizational citizenship behavior via felt responsibility for constructive change; furthermore, the effect of narcissism on change-oriented organizational citizenship behavior via felt responsibility for constructive change would be stronger when the environmental uncertainty prompted by the COVID-19 pandemic was high rather than low. Two studies were conducted to test these hypotheses: an online survey of 180 employees in mainland China (Study 1) and a field study of 167 leader-follower dyads at two Chinese companies (Study 2). The current research reveals a bright side of narcissism, which has typically been recognized as a dark personality trait, and enriches the understanding of the antecedents of change-oriented organizational citizenship behavior. This research can also guide organizations that wish to stimulate employee proactivity.

[Effect of acupuncture at the anterior oblique parietotemporal line on gray matter remodeling in patients with hemiplegia of cerebral infarction].

OBJECTIVE: To observe the intervention effect of acupuncture at the anterior oblique parietotemporal line on gray matter remodeling in patients with hemiplegia of cerebral infarction based on voxel-based morphology (VBM), and to reveal the advantages and central effect sites of scalp acupuncture for hemiplegia of cerebral infarction. METHODS: A total of 18 patients with hemiplegia of cerebral infarction in the right basal ganglia and 18 healthy subjects were enrolled and T1 structural scan was performed. The patients were randomly divided into scalp acupuncture group and non-scalp acupuncture group, with 9 patients in each group. SPSS20.0 was used to analyze the degree of neurological deficit (NIHSS) before and after treatment, and SPM8 software package was used to compare the change in gray matter after treatment between the two groups, the difference in gray matter between patients and healthy subjects. RESULTS: After treatment, the scalp acupuncture group had a significant reduction in NIHSS (P<0.05), while the non-scalp acupuncture group had no significant change in NIHSS (P>0.05); the scalp acupuncture group had a significantly lower NIHSS than the non-scalp acupuncture group after treatment (P<0.05). Compared with the healthy subjects, the patients with cerebral infarction had a voxel increase of gray matter in the bilateral cerebellum; after 2 weeks of treatment, the scalp acupuncture group had a voxel increase of gray matter in the motor-related cortex (such as the cerebellum, the anterior cingulate, the basal nucleus, and the medial frontal gyrus) and some parts of the sensory cortex, while the non-scalp acupuncture group had a slight voxel increase of gray matter in few brain regions which were not associated with motor. After treatment, the scalp acupuncture group had a voxel increase in the brain regions of the basal ganglia, mainly the lenticular nucleus and the claustrum. Compared with the non-scalp acupuncture group after treatment, the scalp acupuncture group had a greater gray matter increase in the brain regions mainly in the bilateral cerebellar cortex. CONCLUSION: Acupuncture at the anterior oblique parietotemporal line can promote neurological function recovery in patients with hemiplegia, possibly by promoting gray matter remodeling in the motor centers of the extrapyramidal system (mainly the bilateral cerebellum and the contralateral basal ganglia) and some parts of the sensory cortex and inducing functional compensation in corresponding brain regions.

Quantitative immunochemical fecal occult blood test for neoplasia in colon cancer screening.

OBJECTIVE: To investigate the performance of the quantitative immunochemical fecal occult blood test (qFIT) and to determine the association between the fecal hemoglobin (Hb) level and the location and size of adenomas and the stages of colorectal cancer (CRC). METHODS: A total of 692 participants were included in the study. Their fecal Hb level was measured using an OC-SENSA MICRO qFIT. The colonoscopy results, including the location, size, and histological features of the adenomas, as well as the relationship between the Hb level and different characteristics were analyzed. Performance of the qFIT at various thresholds of fecal Hb levels was evaluated. RESULTS: Advanced colorectal neoplasia (ACRN) was identified in 76 patients based on the colonoscopic and pathological examinations. Large adenomas (≥10 mm) had a higher fecal Hb level than small adenomas (<10 mm). Advanced adenomas located on the left side of the colon presented with a higher fecal Hb level than those on the right side (P = 0.022). Stage III-IV CRC patients had a significantly higher Hb level than stage I-II patients (P = 0.013). The sensitivity and specificity of qFIT for ACRN was 51.3% and 86.4%，respectively, with the best cut-off level of 400 ng/mL. The sensitivity and specificity for CRC was 61.0% and 89.1%, with the best cut-off level of 500 ng/mL. CONCLUSIONS: qFIT has an acceptable sensitivity and specificity for ACRN detection. Furthermore, the qFIT results are associated with the location and size of adenomas as well as the grade of CRC.

Effect of emodin on chondrocyte viability in an in vitro model of osteoarthritis.

Emodin is an anthraquinone isolated from the Chinese herb Radix et Rhizoma Rhei and has been used to treat various diseases for centuries. The aim of the present study was to investigate the effect of emodin on the inflammatory mediators in rat chondrocytes. In the present study, chondrocytes were isolated from rats, cultured and harvested when they reached generation P3. Cells were treated with different doses of emodin (10, 20, and 30 µg/ml) followed by interleukin 1ß (IL-1ß, 10 ng/ml). Control cells were either untreated or treated with IL-1ß alone. An enzyme-linked immunosorbent assay was used to measure levels of nitric oxide (NO) and prostaglandin E2 (PGE2). Reverse transcription-quantitative polymerase chain was performed to measure levels of matrix metallopeptidase (MMP)-3 and -13 mRNA. The expression of MMP-3, MMP-13, extracellular-signal regulatory kinase (ERK)1/2, phosphorylated ERK1/2 and ß-catenin proteins were detected by western-blot analysis. The results demonstrated that treatment with emodin treatment reduced the cytotoxicity of IL-1ß and inhibited the expression of NO and PGE2 in rat chondrocytes. Furthermore, emodin inhibited the expression of MMP3 and MMP13, and the phosphorylation of various proteins involved in the ERK and Wnt/ß-catenin pathway. Therefore, emodin is able to promote the proliferation of chondrocytes by inhibiting the ERK and Wnt/ß-catenin pathway and downregulating the expression of a series of inflammatory mediators in chondrocytes.

JNK pathway mediates curcumin-induced apoptosis and autophagy in osteosarcoma MG63 cells.

Human osteosarcoma is a common primary malignancy of the bone in children and adolescents. It has been reported that curcumin is able to induce apoptosis in osteosarcoma MG63 cells through the mitochondrial pathway. However, whether curcumin is able to induce autophagy and the interaction between apoptosis and autophagy in osteosarcoma cells has yet to be fully elucidated. In the current study, it was determined that curcumin was able to significantly induce apoptosis, and lead to autophagy in MG63 cells. Notably, inhibition of apoptosis enhanced curcumin-induced autophagy due to upregulation of the c-Jun N-terminal kinase (JNK) signaling pathway. This finding was confirmed by the use of JNK-specific inhibitor, SP600125. Furthermore, our data showed that curcumin-induced apoptosis was increased when autophagy was completely inhibited by 3-methyladenine in MG63 cells. These results suggest that autophagy may have an important role in resistance to apoptosis when MG63 cells are incubated with curcumin. Thus, these results provide important insights into the interaction between apoptosis and autophagy in osteosarcoma cells and clinical treatment strategies using curcumin.

[Space-Time Estimations and Mapping of PM2.5 Fine Particulates Based on Multi-source Data].

PM2.5 pollution in China has become an extreme environmental and social problem and has generated widespread public concern. We estimate ground-level PM2.5 from satellite-derived aerosol optical depth (AOD), topography data, meteorological data, and pollutant emissions using a new technique, Bayesian maximum entropy (BME) combined with geographically weighted regression (GWR), to evaluate the spatial and temporal characteristics of PM2.5 exposure in an eastern region of China in winter. The overall 10-fold cross-validation R2 is 0.92, and the root mean squared prediction error (RMSE) is 8.32 µg·m-3. The mean prediction error (MPE) of the predicted monthly PM2.5 is -0.042 µg·m-3, the mean absolute prediction error (MAE) is 4.60 µg·m-3. Compared with the results of the Geographically Weighted Regression model-GWR (R2=0.71, RMSE=15.68 µg·m-3, MPE=-0.095 µg·m-3, MAE=11.14 µg·m-3), the prediction by the BME were greatly improved. In this location, the high PM2.5concentration area is mainly concentrated in North China, the Yangtze River Delta, and Sichuan Basin. The low concentration area is mainly concentrated in the south of China, including the Pearl River Delta and southwest of Yunnan. Temporally, there is migration trend from the coastal areas inland, and PM2.5 pollution is most serious in December 2015 and January 2016. It is relatively low in November 2015 and February 2016.

Effect of Sinomenine on Expression of COX2, α7nAChR and A2A in A549 Cells / 广州中医药大学学报

Objective To observe the effect of sinomenine (SIN) on the expression of cyclooxygenase (COX2),alpha-7 nicotinic acetylcholine receptor(α7nAChR) and adenosine receptor(A2A) in A549 cells,and to explore the relative mechanism for cell proliferation.Methods The effect of SIN and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on the proliferation of A549 cells was determined by methyl thiazolyl tetrazolium (MTT) assay.The effect of SIN and NNK on the migration of A549 cells was detected by cell wound scratch assay.The effect of SIN and NNK on COX2 expression in A549 cells was determined by Western blotting method.The effect of SIN and NNK on the expression of α7nAChR and A2A mRNA and protein was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting method.Results NNK increased the proliferation and migration of A549 cells,while SIN inhibited the proliferation and migration of A549 cells.COX2 expression level was increased in NNK group but was decreased in SIN group.The expression levels of α7nAChR and A2A were up-regulated in NNK group but were down-regulated in SIN group.Conclusion SIN plays a role in inhibiting the proliferation and migration of A549 cells by suppressing COX2 expression.SIN has an inhibitory effect on the expression of α7nAChR and A2A.

Comparison of pharmacokinetics and subacute toxicity for four arsenic species in rats / 中国食品卫生杂志

Objective Study on the absorption,distribution,transformation,excretion and toxicity of four forms of arsenic in rats.Methods Four thousand and fifty data were obtained from 28 days animal metabolism experiments of 81 rats exposed to the four As species.Distribution,metabolism,excretion,and subacute toxicity of 4 As species were compared by analyzing the arsenate [As(V)],arsenite [As(Ⅲ)],methyl arsenate (MMAV) and dimethyl arsenate (DMAV) content of rat feces (excrement and urine),blood,seven organs,and liver and kidney pathology.Results After oral administration of each As species,82.9％ of As(Ⅲ),85.1％ of As(V),95.0％ of MMAV and 96.2％ of DMAV were accumulatively secreted via feces and urine,while 16.2％ of As(Ⅲ),14.1％ of As(V),4.65％ of MMAV and 0.120％ of DMAV were detected in blood.The DMAV accumulated in blood and urine after dosing As(Ⅲ),As(V) and MMAV,and the content of DMAV in the blood was 8 times greater than urine.Four kinds of arsenic had effects on rat liver and kidney function,the influence on liver was acute phase effect while that on renal was cumulative effects.The effect of As(Ⅲ) was the most significant,followed by MMAV.Four kinds of arsenic could cause tissue deformation and inflammatory cells infiltration,but showed no difference between groups.Conclusion The distribution,metabolism,excretion,and toxicity of the four kinds of arsenic in rats were different in varying degrees.The research of health risk for arsenic should consider its different forms.

[Imaging Observation of Scalp Acupuncture on Brain Gray Matter Injury in Stroke Patients with Cerebral Infarction].

OBJECTIVE: To study features of brain gray matter injury in cerebral infarction patients and intervention of scalp acupuncture by using voxel-based morphology. METHODS: A total of 16 cerebral infarction patients were recruited in this study, and assigned to the scalp acupuncture group and the control group, 8 in each group. Another 16 healthy volunteers were recruited as a normal group. All patients received scanning of T1 structure. Images were managed using VBM8 Software package. Difference of the gray matter structure was compared among the scalp acupuncture group, the control group, and the healthy volunteers. RESULTS: Compared with healthy volunteers, gray matter injury of cerebral infarction patients mainly occurred in 14 brain regions such as cingulate gyrus, precuneus, cuneus, anterior central gyrus, insular lobe, and so on. They were mainly distributed in affected side. Two weeks after treatment when compared with healthy volunteers, gray matter injury of cerebral infarction patients in the scalp acupuncture group still existed in 8 brain regions such as bilateral lingual gyrus, posterior cingulate gyrus, left cuneus, right precuneus, and so on. New gray matter injury occurred in lingual gyrus and posterior cingulate gyrus. Two weeks after treatment when compared with healthy volunteers, gray matter injury of cerebral infarction patients in the control group existed in 23 brain regions: bilateral anterior cingulum, caudate nucleus, cuneate lobe, insular lobe, inferior frontal gyrus, medial frontal gyrus, precuneus, paracentral lobule, superior temporal gyrus, middle temporal gyrus, lingual gyrus, right postcentral gyrus, posterior cingulate gyrus, precentral gyrus, middle frontal gyrus, and so on. New gray matter injury still existed in 9 cerebral regions such as lingual gyrus, posterior cingulate gyrus, postcentral gyrus, and so on. CONCLUSIONS: Brain gray matter structure is widely injured after cerebral infarction. Brain gray matter volume gradually decreased as time went by. Combined use of scalp acupuncture might inhibit the progression of gray matter injury more effectively.

Effect of Sinomenine on Expression of Purinergic Receptors A2A and P2X7 in Mouse Model and In-vitro Macrophages Stimulated by Lipopolysaccharide / 广州中医药大学学报

Objective To investigate the effect of sinomenine on the purinergic receptors A2A and P2X7 in endotoxemia mouse model and RAW264.7 macrophage model stimulated by lipopolysaccharide(LPS). Methods BALB/c mice were randomly divided into blank control group, model group and sinomenine group. Thirty minutes after the rats of sinomenine group were pretreated with intraperitoneal injection of sinomenine (40, 80, 160 mg/kg), the mice were given intraperitoneal injection of 15 mg/kg LPS to induce endotoxemia model. The serum levels of tumor necrosis factor-alpha(TNF-α) and interleukin-6(IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). The expression levels of purinergic receptor A2A and P2X7 in the liver and spleen were detected by reverse transcription-polymerase chain reaction(RT-PCR). RAW264.7 macrophages were divided into blank control group, LPS group and sinomenine group. Sinomenine group was firstly treated with sinomenine(300μmol/L) for 2 h, and then LPS group and sinomenine group were treated with LPS (100 ng/mL) for another 8 hours. TNF-α in the cell supernatant was measured by ELISA, and the expression levels of A2A and P2X7 in RAW264.7 cells were detected by RT-PCR. Results Stimulation with LPS could induce the increase of the mouse serum levels of TNF-α and IL-6 as well as the expression of A2A and P2X7 in mouse liver and spleen, and sinomenine had a counteraction on the above indexes(P<0.05) . In-vitro stimulation with LPS could induce the increase of the content of TNF-α and the expression of A2A and P2X7 in RAW264.7 cells , and sinomenine decreased TNF-α content and P2X7 expression (P<0.05) , but had an effect on enhancing A2A expression. Conclusion Sinomenine suppresses the expression of purinergic receptor P2X7 in mouse spleen and liver as well as in RAW264.7 macrophages, but its effect on the expression of A2A in various tissues and cells varies, whose related mechanism is needed further study.

Covalent bonding of YIGSR and RGD to PEDOT/PSS/MWCNT-COOH composite material to improve the neural interface.

The development of coating materials for neural interfaces has been a pursued to improve the electrical, mechanical and biological performances. For these goals, a bioactive coating was developed in this work featuring a poly(3,4-ethylenedioxythiophene) (PEDOT)/carbon nanotube (CNT) composite and covalently bonded YIGSR and RGD. Its biological effect and electrical characteristics were assessed in vivo on microwire arrays (MWA). The coated electrodes exhibited a significantly higher charge storage capacity (CSC) and lower electrochemical impedance at 1 kHz which are desired to improve the stimulating and recording performances, respectively. Acute neural recording experiments revealed that coated MWA possess a higher signal/noise ratio capturing spikes undetected by uncoated electrodes. Moreover, coated MWA possessed more active sites and single units, and the noise floor of coated electrodes was lower than that of uncoated electrodes. There is little information in the literature concerning the chronic performance of bioactively modified neural interfaces in vivo. Therefore in this work, chronic in vivo tests were conducted and the PEDOT/PSS/MWCNT-polypeptide coated arrays exhibited excellent performances with the highest mean maximal amplitude from day 4 to day 12 during which the acute response severely compromised the performance of the electrodes. In brief, we developed a simple method of covalently bonding YIGSR and RGD to a PEDOT/PSS/MWCNT-COOH composite improving both the biocompatibility and electrical performance of the neural interface. Our findings suggest that YIGSR and RGD modified PEDOT/PSS/MWCNT is a promising bioactivated composite coating for neural recording and stimulating.

Effect of sinomenine on mouse RAW264.7 macrophage cells line polarization induced by LPS or IL-4 / 中国免疫学杂志

Objective:To investigate sinomenine (Sinomenine,SIN) effect on RAW264.7 cells polarization to M1 or M2 phenotype induced by lipopolysaccharide (LPS) or interleukin-4 (IL-4) .Methods:RAW264.7 cells were induced to polarize to M1 by LPS ,and to M2 by IL-4.Sinomenine effects on LPS or IL-4 induced macrophages:TNF-αand IL-10 secretion induced by different condition were detected by Enzyme linked immunosorbent assay (ELISA);The expression level of mRNA of Arginase1(Arg-1),Nitric oxide synthase(iNOS),suppressor of cytokine signaling protein-2(SOCS2) and suppressor of cytokine signaling protein-3(SOCS3) of M1/M2 phenotypes were detected by real time PCR respectively.Results:Sinomenine inhibited the increase of TNF-αsecretion,iNOS and SOCS3 mRNA expression level induced by LPS.Sinomenine inhibited the increase of IL-10 secretion and Arg-1 mRNA expression level induced by IL-4,but SOCS2 mRNA expression level was not affected by Sinomenine.Conclusion: Sinomenine can inhibite the macrophage polarization to M1 and M2 induced by LPS and IL-4.Sinomenine plays a regulatory role on imbalance of M1/M2,and is conducive to maintain the dynamic balance.