Activation of TRPV1 receptor facilitates myelin repair following demyelination via the regulation of microglial function.

The transient receptor potential vanilloid 1 (TRPV1) is a non-selective cation channel that is activated by capsaicin (CAP), the main component of chili pepper. Despite studies in several neurological diseases, the role of TRPV1 in demyelinating diseases remains unknown. Herein, we reported that TRPV1 expression was increased within the corpus callosum during demyelination in a cuprizone (CPZ)-induced demyelination mouse model. TRPV1 deficiency exacerbated motor coordinative dysfunction and demyelination in CPZ-treated mice, whereas the TRPV1 agonist CAP improved the behavioral performance and facilitated remyelination. TRPV1 was predominantly expressed in Iba1+ microglia/macrophages in human brain sections of multiple sclerosis patients and mouse corpus callosum under demyelinating conditions. TRPV1 deficiency decreased microglial recruitment to the corpus callosum, with an associated increase in the accumulation of myelin debris. Conversely, the activation of TRPV1 by CAP enhanced the recruitment of microglia to the corpus callosum and potentiated myelin debris clearance. Using real-time live imaging we confirmed an increased phagocytic function of microglia following CAP treatment. In addition, the expression of the scavenger receptor CD36 was increased, and that of the glycolysis regulators Hif1a and Hk2 was decreased. We conclude that TRPV1 is an important regulator of microglial function in the context of demyelination and may serve as a promising therapeutic target for demyelinating diseases such as multiple sclerosis.

Comparative Transcriptome Analysis of Hard and Tender Fruit Spines of Cucumber to Identify Genes Involved in the Morphological Development of Fruit Spines.

The spines of cucumber fruit not only have important commercial value but are also a classical tissue to study cell division and differentiation modes of multicellular trichomes. It has been reported that CsTs (C-type Lectin receptor-like kinase) can influence the development of fruit spines. In this study, we took a pair of cucumber materials defined as hard (Ts, wild type) and tender spines (ts, mutant) and defined the developmental process of fruit spines as consisting of four stages (stage I to stage IV) by continuously observing by microscope and SEM. Comparisons of transcriptome profiles at different development stages of wild-type spines showed that 803 and 722 genes were upregulated in the stalk (stage II and stage III) and base (stage IV) development stages of fruit spines, respectively. The function analysis of DEGs showed that genes related to auxin polar transport and HD-ZIP transcription factor are significantly upregulated during the development of the stalk. bHLH transcription factors and cytoskeleton-related genes were significantly upregulated during the development of the base. In addition, stage III is the key point for the difference between wild-type and mutant spines. We detected 628 DEGs between wild type and mutant at stage III. These DEGs are mainly involved in the calcium signaling of the cytoskeleton and auxin polar transport. Coincidentally, we found that CsVTI11, a factor involved in auxin signal transmission, can interact with CsTs in vivo, but this interaction does not occur between CsVTI11 and Csts, further suggesting that CsTs may regulate the development of fruit spines by influencing cell polarity. These results provide useful tools to study the molecular networks associated with cucumber fruit spine development and elucidate the biological pathways that C-type Lectin receptor-like kinase plays in regulating the development of fruit spines.

A specialized myodural bridge named occipital-dural muscle in the narrow-ridged finless porpoise (Neophocaena asiaeorientalis).

A dense bridge-like tissue named the myodural bridge (MDB) connecting the suboccipital muscles to the spinal dura mater was originally discovered in humans. However, recent animal studies have revealed that the MDB appears to be an evolutionarily conserved anatomic structure which may have significant physiological functions. Our previous investigations have confirmed the existence of the MDB in finless porpoises. The present authors conducted research to expound on the specificity of the MDB in the porpoise Neophocana asiaeorientalis (N.asiaeorientalis). Five carcasses of N.asiaeorientalis, with formalin fixation, were used for the present study. Two of the carcasses were used for head and neck CT scanning, three-dimensional reconstructions, and gross dissection of the suboccipital region. Another carcass was used for a P45 plastination study. Also, a carcass was used for a histological analysis of the suboccipital region and also one was used for a Scanning Electron Microscopy study. The results revealed that the MDB of the N.asiaeorientalis is actually an independent muscle originating from the caudal border of the occiput, passing through the posterior atlanto-occipital interspace, and then attaches to the cervical spinal dura mater. Thus the so called MDB of the N.asiaeorientalis is actually an independent and uniquely specialized muscle. Based on the origin and insertion of this muscle, the present authors name it the 'Occipital-Dural Muscle'. It appears that the direct pull of this muscle on the cervical spinal dura mater may affect the circulation of the cerebrospinal fluid by altering the volume of the subarachnoid space via a pumping action.

Anti-inflammatory monoterpene esters from the stems of Illigera aromatica.

Two new monoterpene esters illigerates F and G (1 and 2) together with 5 know compounds illigerate A (3), illigerate C (4), actinodaphnine (5), N-methylactinodaphnine(6) and N-methyllaurotetanine(7) were isolated from Illigera aromatica S. Z. Huang et S. L. Mo. Their structures were identified by extensive NMR data and by comparing with the known compounds. The anti-inflammatory activity of four monoterpenes (1 - 4) was evaluated by inhibiting nitric oxide (NO) production in lipopolysaccharide-activated murine macrophage RAW 264.7 cells and four monoterpenoids exhibited inhibitory effect with IC50 values of 71.5 ± 7.3, 74.7 ± 5.6, 48.0 ± 7.4 and 65.1 ± 3.7 µM, respectively.

Genome-Wide Identification and Characterization of Lectin Receptor-Like Kinase Gene Family in Cucumber and Expression Profiling Analysis under Different Treatments.

Lectin receptor-like kinases (LecRLKs) are a class of membrane proteins found in plants that are involved in diverse functions, including plant development and stress responses. Although LecRLK families have been identified in a variety of plants, a comprehensive analysis has not yet been undertaken in cucumber (Cucumis sativus L.). In this study, 46 putative LecRLK genes were identified in the cucumber genome, including 23 G-type and 22 L-type, and one C-type LecRLK gene. They were unequally distributed on all seven chromosomes, with a clustering tendency. Most of the genes in the cucumber LecRLK (CsLecRLK) gene family lacked introns. In addition, there were many regulatory elements associated with phytohormones and stress on these genes' promoters. Transcriptome data demonstrated distinct expression patterns of CsLecRLK genes in various tissues. Furthermore, we found that each member of the CsLecRLK family had its own unique expression pattern under hormone and stress treatment by the quantitative real-time PCR (qRT-PCR) analysis. This study provides a better understanding of the character and function of the LecRLK gene family in cucumber and opens up the possibility to exploring the roles that LecRLKs might play in the life cycle of cucumber.

Scanning Electron Microscopic Observation of Myodural Bridge in the Human Suboccipital Region.

STUDY DESIGN: A scanning electron microscopic study performed on three cadaveric specimens focused on the human suboccipital region, specifically, myodural bridge (MDB). OBJECTIVE: This study showed the connection form of the MDB among the suboccipital muscles, the posterior atlanto-occipital membrane (PAOM) and the spinal dura mater (SDM), and provided an ultrastructural morphological basis for the functional studies of the MDB. SUMMARY OF BACKGROUND DATA: Since the myodural bridge was first discovered by Hack, researches on its morphology and functions had been progressing continuously. However, at present, research results about MDB were still limited to the gross anatomical and histological level. There was no research report showing the MDB's ultrastructural morphology and its ultrastructural connection forms between PAOM and SDM. METHODS: A scanning electron microscope (SEM) was used to observe the connection of myodural bridge fibers with PAOM and SDM in atlanto-occipital and atlanto-axial interspaces, and the connection forms were analyzed. RESULTS: Under the SEM, it was observed that there were clear direct connections between the suboccipital muscles and the PAOM and SDM in the atlanto-occipital and atlanto-axial spaces. These connections were myodural bridge. The fibers of the myodural bridge merged into the spinal dura mater and gradually became a superficial layer of the spinal dura mater. CONCLUSION: MDB fibers merged into the SDM and became part of the SDM in the atlanto-occipital and atlanto-axial space. MDB could transfer tension and pulling force to the SDM effectively, during the contraction or relaxation of the suboccipital muscles. LEVEL OF EVIDENCE: N/A.

[Electroacupuncture promotes regeneration and repair of myelin sheath of corpus callosum in demyelination mice].

OBJECTIVE: To explore the mechanism of electroacupuncture (EA) in accelerating the aggregation of microglia and promoting the remyelination at the location of demyelination. METHODS: C57BL/6 mice were randomly divided into 4 groups: normal, control, model (LPC) and LPC＋EA. The demyelination model was established by microinjection of Lysolecithin (LPC, 1 µL) into the left corpus callosum. EA (2 Hz/15 Hz, 2－4 mA) was applied to "Baihui"(GV20)and "Zhiyang"(GV9)for 30 min，once daily for 3 days, then, once every other day for 18 days. Immuno-fluorescence staining was used to observe the expression of myelin basic protein (MBP) and Axl tyrosine kinase receptor (Axl), Iba1 and numbers of Olig2-positive oligodendrocytes in the corpus callosum. Western blot was employed to detect the expression of MBP in the corpus callosum, and Oil Red O staining was used to observe changes of number of myelin pieces. RESULTS: Following modeling, the expression levels of MBP on day 5 and 10 after modeling were significantly decreased (P<0.05, P<0.01), Iba1 expression and Olig2-positive oligodendrocyte numbers on day 10 apparently increased (P<0.001, P<0.01). On day 21 after modeling, the levels of the above mentioned indexes returned to normal. After EA intervention, the levels of MBP expression on day 5 and 10, Axl, Iba1 protein expression and Olig2-positive oligodendrocyte numbers on day 5 were markedly increased (P<0.001，P<0.01，P<0.05), while Iba1 expression on day 10 was considerably decreased in comparison with the model group (P<0.01)．Oil Red O staining showed that on day 5 after modeling, the number of red lipid droplets were obviously increased in the corpus callosum tissue on the injection side, and apparently reduced in the EA group, suggesting a clearance of the accumulated myelin fragments by EA. CONCLUSION: EA intervention may reduce myelin debris and promote the aggregation of microglial cells and oligodendrocytes to the injured site, accelerate the myelin regeneration and up-regulate the expression of MBP and Axl of corpus callosum in demyelination mice.

miR-124/VAMP3 is a novel therapeutic target for mitigation of surgical trauma-induced microglial activation.

Activation of microglia and the subsequently elevated inflammatory cytokine release in the brain during surgery predispose individuals to cognitive dysfunction, also known as postoperative cognitive dysfunction (POCD). miR-124 is one of the most abundant microRNAs in the brain that regulates microglial function. Elucidating the role of miR-124 in microglial activation in the context of surgery may therefore promote understanding of as well as therapeutic development for post-surgical disorders involving microglial activation. The downstream targets of miR-124 were investigated using bioinformatic screening and dual-luciferase reporter assay validation, and vesicle-associated membrane protein 3 (VAMP3) was identified as a potential target. The kinetics of miR-124/VAMP3 expression was first examined in vitro in microglial cells (primary microglia and BV2 microglial cells) following lipopolysaccharide (LPS) stimulation. LPS induced a time-dependent decrease of miR-124 and upregulated the expression of VAMP3. Manipulating miR-124/VAMP3 expression by using miR-124 mimics or VAMP3-specific siRNA in LPS-stimulated BV2 microglial cells inhibited BV2 microglial activation-associated inflammatory cytokine release. To further examine the role of miR-124/VAMP3 in a surgical setting, we employed a rat surgical trauma model. Significant microglial activation and altered miR-124/VAMP3 expression were observed following surgical trauma. We also altered miR-124/VAMP3 expression in the rat surgical trauma model by administration of exogenous miR-124 and by using electroacupuncture, which is a clinically applicable treatment that modulates microglial function and minimizes postoperative disorders. We determined that electroacupuncture treatment specifically increases the expression of miR-124 in the hypothalamus and hippocampus. Increased miR-124 expression with a concomitant decrease in VAMP3 expression resulted in decreased inflammatory cytokine release related to microglial activation post-surgery. Our study indicates that miR-124/VAMP3 is involved in surgery-induced microglial activation and that targeting miR-124/VAMP3 could be a potential therapeutic strategy for postoperative disorders involving microglial activation.

Three new diterpenoids from Aralia dumetorum.

Three new diterpenoids, dumetoranes A (1) and B (2), melanocane B (3), together with four known ones including melanocane A (4), ent-15S,16-dihydroxypimar-8(14)-en-19-oic acid (5), ent-pimara-8(14),15-diene-19-oic acid (6), and ent-pimara-8(14),15-diene-19-ol (7) were obtained from the ethanol extract of the roots of Aralia dumetorum. Their structure elucidation was achieved by the methods of spectroscopic HRMS, IR, NMR, and by comparison with literature. The cytotoxicities of compounds 1-3 and 5 were assayed by in vitro MTT methods.

α-Glucosidase Inhibitory and Anti-Inflammatory Coumestans from the Roots of Dolichos trilobus.

Four new coumestans dolichosins A - D (1: -4: ) were isolated from the roots of Dolichos trilobus, together with four known compounds: isosojagol (5: ), phaseol (6: ), psoralidin (7: ), and 4″,5″-dehydroisopsoralidin (8: ). Their structures were elucidated on the basis of spectroscopic data interpretation, mass spectrometric analyses, and the comparison with literature data of related compounds. The anti-inflammatory activity of these compounds (1: -8: ) was evaluated through the inhibition of nitric oxide production in lipopolysaccharide-activated murine macrophage RAW 264.7 cells, in which compounds 1: and 6: displayed moderate inhibitory activity and no cytotoxic effects. In a α-glucosidase inhibitory assay, compounds 1: and 5: -8: exhibited appreciable inhibition on α-glucosidase. Especially compounds 1, 7: , and 8: showed IC50 values lower than 20.0 µM.

Chemical constituents from the whole herb of Hemiphragma heterophyllum.

Phytochemical investigation on Hemiphragma heterophyllum led to the isolation of two new compounds, heterophyllumin A (1) and heterophylliol (3), along with nine known compounds, (‒)-sibiricumin A (2), iridolactone (4), jatamanin A (5), dihydrocatalpolgenin (6), 25-hydroperoxycycloart-23-en-3ß-ol (7), 24-methylenecycloartanol (8), (+)-pinoresinol (9), hexadec-(4Z)-enoic acid (10), and 9,12, 15-octadecatrienoic acid (11). Their structures were elucidated on the basis of detailed spectroscopic analyses and by comparison with literature data. Further, the structure of compound 3 was unambiguously confirmed by single-crystal X-ray analysis. Some of those compounds showed moderate activity in the α-glucosidase inhibition assay.

Effect of local wound infiltration with ropivacaine on postoperative pain relief and stress response reduction after open hepatectomy.

AIM: To prospectively evaluate the effect of local wound infiltration with ropivacaine on postoperative pain relief and stress response reduction after open hepatectomy. METHODS: A total of 56 patients undergoing open hepatectomy were randomly divided into two groups: a ropivacaine group (wound infiltration with ropivacaine solution) and a control group (infiltration with isotonic saline solution). A visual analog scale (VAS) at rest and on movement was used to measure postoperative pain for the first 48 h after surgery. Mean arterial pressure (MAP), heart rate (HR), time to bowel recovery, length of hospitalization after surgery, cumulative sufentanil consumption, and incidence of nausea and vomiting were compared between the two groups. Surgical stress hormones (epinephrine, norepinephrine, and cortisol) were detected using enzyme-linked immunosorbent assay, and the results were compared. RESULTS: VAS scores both at rest and on movement at 24 h and 48 h were similar between the two groups. Significantly lower VAS scores were detected at 0, 6, and 12 h in the ropivacaine group compared with the control group (P < 0.05 for all). MAP was significantly lower at 6, 12, and 24 h (P < 0.05 for all); HR was significantly lower at 0, 6, 12, and 24 h (P < 0.05 for all); time to bowel recovery and length of hospitalization after surgery (P < 0.05 for both) were significantly shortened; and cumulative sufentanil consumption was significantly lower at 6, 12, 24, and 36 h (P < 0.05 for all) in the ropivacaine group than in the control group, although the incidence of nausea and vomiting showed no significant difference between the two groups. The levels of epinephrine, norepinephrine, and cortisol were significantly lower in the ropivacaine group than in the control group at 24 and 48 h (P < 0.01 for all). CONCLUSION: Local wound infiltration with ropivacaine after open hepatectomy can improve postoperative pain relief, reduce surgical stress response, and accelerate postoperative recovery.

Acutissimanide, a new lignan with antioxidant activity isolated from the bark of Quercus acutissima Carruth.

Acutissimanide (1), a new lignin, together with 11 known polyphenols (2-12) were isolated from the bark of the deciduous oak tree, Quercus acutissima Carruth. The structure of compound 1 was determined using multidimensional (1)H and (13)C NMR and mass spectroscopy. The antioxidant properties of compounds 1-12 were investigated using a 1,1-diphenyl-2-picryhydrazyl radical-scavenging assay with compounds 6-11 displaying significant antioxidant activity (EC50 values of 5.2-23.7 µM). Our findings suggest the extracts of Q. acutissima Carruth are a potential source of natural antioxidant additives for use in the food and other allied industries.