Therapeutic potential of CB1R activation by Qingyangshen glycoside M1 for seizure relief.

ETHNOPHARMACOLOGICAL RELEVANCE: Cynanchum otophyllum C.K.Schneid.PI.Wilson, commonly referred as ''Qingyangshen'' (QYS), is a traditional folk medicine from Yunnan, renowned for its efficacy in neurological and psychiatric disorders. Glycosides isolated from QYS have shown promise in alleviating epilepsy, however, mechanisms of action and specific molecular targets remain to be elucidated. AIM OF THE STUDY: The study aimed to evaluate the anticonvulsant effects of Qingyangshen glycosides M1 (M1), a C21 steroidal glycoside from QYS, on pentylenetetrazol (PTZ)-induced convulsions in zebrafish (Danio rerio), and its neuroprotective effect on Glutamate (Glu)-induced damage to PC12 cells, and importantly to identify its potential molecular targets. MATERIALS AND METHODS: To evaluate anticonvulsant activity of M1, 7 days-post-fertilization (7-dpf) animals were pretreated (by immersion) and then exposed to PTZ (10 mM) solution. Furthermore, Glu-induced PC12 cell damage was employed to investigate the neuroprotective and anti-apoptotic capacity. Cells were pretreated with various concentrations of M1 (0-10 µM) for 12 h and then co-treated with Glu (15 mM) for an additional 24 h. The cell viability, apoptosis rate and apoptosis-related proteins (p-PI3K, PI3K, Akt, p-Akt, CREB, p-CREB, BDNF, Bax and Bcl-2) were measured using CCK-8, annexin V/PI and Western blot assays. To model the expected interaction between M1 and candidate cannabinoid receptor type 1 (CB1R), ERK phosphorylation, molecular docking, and drug affinity responsive target stability (DARTS) techniques were employed. Finally, CB1R antagonist Rimonabant (Rim) was validated by co-administration in both zebrafish and cells to confirm the requirement of CB1R for M1 efficacy. RESULTS: At a concentration of 400 µM, M1 dramatically reversed PTZ-induced convulsive-like behaviors in zebrafish, as evidenced by a significant reduction in locomotor activity. In the context of Glu-induced cytotoxicity, M1 (10 µM) demonstrated a notable increase in cell viability and suppressed apoptosis through modulation of the Bax/Bcl-2 ratio and activation of the PI3K/Akt/CREB/BDNF signaling axis. These effects were facilitated through CB1R activation. In contrast, Rim dampened the beneficial activities of M1 as a cannabinoid agonist. CONCLUSIONS: These results demonstrated that M1 as a potential CB1R activator, exhibiting anticonvulsive effects in a PTZ-induced zebrafish model and neuroprotective properties via the PI3K/Akt/CREB/BDNF signaling axis in a Glu-induced PC12 cell injury model. Notably, the observed seizure relief attenuated by CB1R chemical antagonism.

Iristectorigenin C suppresses LPS-induced macrophages activation by regulating mPGES-1 expression and p38/JNK pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Nonsteroidal anti-inflammatory drugs (NSAIDs) have been used clinically to treat inflammatory diseases clinically. However, the adverse effects of NSAIDs cannot be ignored. Therefore, it is critical for us to find alternative anti-inflammatory drugs that can reduce adverse reactions to herbal medicine, such as Iris tectorum Maxim., which has therapeutic effects and can treat inflammatory diseases and liver-related diseases. AIM OF THE STUDY: This study aimed to isolate active compounds from I. tectorum and investigate their anti-inflammatory effects and action mechanisms. MATERIALS AND METHODS: Fourteen compounds were isolated from I. tectorum using silica gel column chromatography, Sephadex LH-20, ODS and high performance liquid chromatography, and their structures were identified by examining physicochemical properties, ultraviolet spectroscopy, infrared spectroscopy, mass spectrometry, and nuclear magnetic resonance spectroscopy. Classical inflammatory cell models were established using lipopolysaccharide (LPS)-stimulated RAW264.7 cells and rat primary peritoneal macrophages to examine the effect of these compounds. To examine the action mechanisms, the nitric oxide (NO) levels were measured by Griess reagent and the levels of inflammatory cytokines in the supernatant were measured by ELISA; The expressions of major proteins in prostaglandin E2 (PGE2) synthesis and the nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways were examined by Western blotting, and the mRNA expression levels were measured by quantitative real-time polymerase chain reaction; and the nuclear translocation of p65 was examined by high content imaging. Molecular docking was used to predict the binding of active compound to target protein. RESULTS: Our findings revealed that Iristectorigenin C (IT24) significantly inhibited the levels of NO and PGE2 without affecting cyclooxygenase (COX)-1/COX-2 expression in LPS-induced RAW264.7 cells and rat peritoneal macrophages. Furthermore, IT24 was shown to decrease the expression of microsomal prostaglandin synthetase-1 (mPGES-1) in LPS-induced rat peritoneal macrophages. IT24 did not suppress the phosphorylation and nuclear translocation of proteins in the NF-κB pathway, but it inhibited the phosphorylation of p38/JNK in LPS-stimulated RAW264.7 cells. Additionally, molecular docking analysis indicated that IT24 may directly bind to the mPGES-1 protein. CONCLUSION: IT24 might inhibit mPGES-1 and the p38/JNK pathway to exert its anti-inflammatory effects and could be also developed as an inhibitor of mPGES-1 to prevent and treat mPGES-1-related diseases, such as inflammatory diseases, and holds promise for further research and drug development.

Glutathione-modified graphene quantum dots as fluorescent probes for detecting organophosphorus pesticide residues in Radix Angelica Sinensis.

A novel fluorescent sensor was developed in this study based on glutathione-functionalized graphene quantum dots (GQDs@GSH) to detect organophosphorus pesticide residues in Radix Angelica Sinensis. GQDs@GSH was synthesized by a one-step pyrolysis method with a fluorescence quantum yield as high as 33.9% and its structure was characterized by transmission electron microscopy and X-ray photoelectron spectroscopy. GQDs@GSH exhibited excellent fluorescence property showing strong blue fluorescence under UV irradiation. The fluorescence of GQDs@GSH could be quenched by Fe3+ by electron transfer and the quenched fluorescence could be recovered due to the strong chelating and reducing ability of phytic acid (PA). Under the catalyzation of acetylcholinesterase (AChE) and choline oxidase (ChOx), acetylcholine (ACh) could be decomposed to H2O2, which could further oxidize Fe2+ to Fe3+ thus quenching the fluorescence of GQDs@GSH once again. Coumaphos, a kind of organophosphorus pesticide, could inhibit AChE activity, thus making the quenched fluorescence turn on again. Several parameters influencing the fluorescence response such as Fe3+, PA, ACh and coumaphos concentration, pH value and reaction time were optimized. Based on such a fluorescence "off-on-off-on" ngkmechanism, GQDs@GSH was successfully applied to the detection of coumaphos in Radix Angelica Sinensis. A good linear relationship between the fluorescence intensity and coumaphos concentration was obtained in the range of 0.1-10.0 µmol·L-1. By a standard addition method, the recoveries were measured to be 101.44-117.90% with RSDs lower than 1.98%. The biosensor system is simple, sensitive and accurate. It has a good application prospect in the detection of organophosphorus pesticide residues in traditional Chinese medicine and agricultural products, and also expanded the application scope for glutathione as a highly selective biological molecule.

Decoding the Key Functional Combined Components Group and Uncovering the Molecular Mechanism of Longdan Xiegan Decoction in Treating Uveitis.

Objective: Longdan Xiegan Decoction (LXD) is a famous herbal formula in China. It has been proved that LXD has been shown to have a significant inhibitory effect on suppresses the inflammatory cells associated with uveitis. However, the key functional combination of component groups and their possible mechanisms remain unclear. Methods: The community detecting model of the network, the functional response space, and reverse prediction model were utilized to decode the key components group (KCG) and possible mechanism of LXD in treating uveitis. Finally, MTT assay, NO assay and ELISA assay were applied to verify the effectiveness of KCG and the accuracy of our strategy. Results: In the components-targets-pathogenic genes-disease (CTP) network, a combination of Huffman coding and random walk algorithm was used and eight foundational acting communities (FACs) were discovered with important functional significance. Verification has shown that FACs can represent the corresponding C-T network for treating uveitis. A novel node importance calculation method was designed to construct the functional response space and pick out 349 effective proteins. A total of 54 components were screened and defined as KCG. The pathway enrichment results showed that KCG and their targets enriched signal pathways of IL-17, Toll-like receptor, and T cell receptor played an important role in the pathogenesis of uveitis. Furthermore, experimental verification results showed that important KCG quercetin and sitosterol markedly inhibited the production of nitric oxide and significantly regulated the level of TNF-α and IFN-γ in Lipopolysaccharide-induced RAW264.7 cells. Discussion: In this research, we decoded the potential mechanism of the multi-components-genes-pathways of LXD's pharmacological action mode against uveitis based on an integrated pharmacology approach. The results provided a new perspective for the future studies of the anti-uveitis mechanism of traditional Chinese medicine.

First report of Alternaria alternata causing leaf blight on Actaea dahurica in China.

In July 2019, leaf blight on Actaea dahurica, a plant with high value in Chinese traditional medicine, was discovered in a 2 ha planting area in Heilongjiang Province (129.6°E, 44.6°N), China. Disease incidence was 90% in the field. Symptoms consisted of irregular black spots with gray margins on both sides of the leaf, often at the leaf margin, mostly on the older leaves. To isolate the pathogen, ten diseased leaves were randomly collected, surface disinfested, and 5 x 5 mm segments were removed from the margin of the lesions. Leaf segments were placed onto potato dextrose agar (PDA) and incubated at 25 ℃ for 7 days. Ten pure cultures with the same morphological characteristics were obtained from three leaves showing typical symptoms. Cultures on PDA initially had a cottony mycelium, white-gray to gray. After two to three weeks of growth, mycelium color changed from gray to black. Conidiophores were clustered, dark at the base, tapering to the apex, born from simple sublates, unbranched, with 1 to 5 septa, and 70.4-530.3 × 5-7.5 µm in size. Conidia were 12.5-82.5 × 5.2-20.3 µm, usually in chains, had 2 to 8 transverse septa, 0 to 4 longitudinal or oblique septa, and a smooth brown surface. Simple, pale, vimineous or verrucous beaks developed from the apical cells with 0 to 4 septa. The morphological characteristics were consistent with Alternaria species (Simmons, 2007). To fulfill Koch's postulates, pathogenicity tests were carried out on three-month-old A. dahurica plants. A spore suspension was prepared from PDA cultures of isolates SM0101 and SM0102 and adjusted to 105 spores/mL using a hemocytometer. Each leaf was sprayed with 2 mL of the spore suspension, then incubated at 25 ℃ for 7 days. The same number of healthy A. dahurica plants were sprayed with sterile water as a control. After 7 days, small brown necrotic spots appeared on inoculated plants, but the control group showed no symptoms. A fungus with the same characteristics as that used for inoculation was re-isolated from the lesions. This experiment was replicated three times, and the results of each experiment were consistent. Genomic DNA was extracted from isolates SM0101 and SM0102 and used for PCR amplification of the rDNA internal transcribed spacer regions (ITS), RNA polymerase II gene (RPB2) and Alternaria allergen a 1 (Alt a 1) gene sequences using the primer pairs ITS1/ITS4 (White et al. 1990), RPB2-5F2/RPB2-7CR (Khodaei and Arzanlou, 2013) and Alt-for/Alt-rev (Hong et al. 2005), respectively. The ITS (OL703042, OL616086), RPB2 (OL703043, OL898416), and Alt a 1 sequences (OL616087, OL898415) were deposited in GenBank. The sequences obtained in this study had the highest match to corresponding sequences of Alternaria alternata CBS 916.96 (AF347031, KC584375, AY563301). For isolate SM0101 the matches were ITS (461/461 bp), RPB2 (897/985 bp), and Alt a 1 (488/488 bp). For isolate SM0202 the matches were ITS (457/457 bp), RPB2 (893/985 bp), and Alt a 1 (484/484 bp). A phylogenetic analysis was performed using MEGA7 software. The alignment included sequences from 16 ex-type Alternaria species and the two isolates causing leaf blight on A. dahurica. Branch supports were calculated with 1,000 bootstrap replicates, and phylogenetic inference was performed using the maximum likelihood estimation. The fungus isolated from A. dahurica clustered with A. alternata. This is the first report of A. alternata on A. dahurica in the world. This report will help to identify the disease symptoms in the field and provides a basis for research into the occurrence, distribution, and control of leaf blight on A. dahurica.

[Current status of organophosphorus pesticide residues in root and rhizome medicinal materials and research progress of rapid detection methods].

The medicinal plants with roots and rhizomes as the medicinal parts account for about 1/3 of Chinese medicinal herbs. Root and rhizome medicinal materials are widely used in clinical practice, whereas their wild resource reserves are insufficient to meet the market demand. With the expansion of planting areas, the formation of large-scale production areas, and the increase in planting years, diseases and insect pests of these medicinal plants, which are diverse and have broad transmission routes, strong concealment, and heavy damage, have become more and more serious. The prevention and control of these diseases and insect pests is characterized by multiple ways of pesticide application, large consumption of pesticides, susceptibility to soil barrier, difficulty in the control, and unstable control efficiency. Organophosphorus pesticides(OPPs) are widely used in the cultivation of Chinese medicinal plants because of their diverse varieties, broad-spectrum, good efficacy, and low residues, and have a positive effect on the yield and quality of Chinese medicinal materials. However, the abuse of OPPs not only increases the planting cost, but also affects the quality and safety of Chinese medicinal plants, the safety of clinical use of Chinese medicine, and the ecological safety of production areas. This paper reviewed the research and development progress of OPPs, the registration status of OPPs used in root and rhizome medicinal materials, residue limit standards, residue status, and rapid detection technology progress of OPPs. This review aims to provide research ideas and references for standardizing the use of OPPs in root and rhizome medicinal materials, reducing OPP residues, and establishing a fast, efficient, accurate, and reliable method for the detection of OPP residues in Chinese herbal medicine.

[ZHUANG Li-xing's experience of using mind-regulation acupuncture for postherpetic neuralgia].

Professor ZHUANG Li-xing's clinical experience of using mind-regulation acupuncture for postherpetic neuralgia is summarized. Professor ZHUANG believes that the physical pain symptoms in patients could cause negative emotions, which often lead to a series of mental symptoms, which in turn aggravate or induce pain. Therefore, the treatment for this disease should start with mind regulation, with Baihui (GV 20) through Sishenzhen, Shenmen (HT 7), Neiguan (PC 6), Shenmai (BL 62), Zhaohai (KI 6), Siguan (Hegu (LI 4) and Taichong (LR 3)), Sanyinjiao (SP 6), Shuigou (GV 26) and Suliao (GV 25) as main acupoints to regulate mind and relieve pain, in combination of local analgesia and ear acupuncture, which has obtained satisfied curative effect.

's experience of using mind-regulation acupuncture for postherpetic neuralgia / 中国针灸

Professor 's clinical experience of using mind-regulation acupuncture for postherpetic neuralgia is summarized. Professor believes that the physical pain symptoms in patients could cause negative emotions, which often lead to a series of mental symptoms, which in turn aggravate or induce pain. Therefore, the treatment for this disease should start with mind regulation, with Baihui (GV 20) through , Shenmen (HT 7), Neiguan (PC 6), Shenmai (BL 62), Zhaohai (KI 6), Siguan (Hegu (LI 4) and Taichong (LR 3)), Sanyinjiao (SP 6), Shuigou (GV 26) and Suliao (GV 25) as main acupoints to regulate mind and relieve pain, in combination of local analgesia and ear acupuncture, which has obtained satisfied curative effect.

Expression pattern and pharmacological characterisation of two novel alternative splice variants of the glutamate-gated chloride channel in the small brown planthopper Laodelphax striatellus.

BACKGROUND: Glutamate-gated chloride channels (GluCl) mediate fast inhibitory neurotransmission in invertebrate nervous systems. Although only one GluCl gene was presented in insects, it showed diverse alternative splicing that was speculated could affect channel function and pharmacology. RESULTS: In this study, we isolated GluCl cDNAs from adults of the small brown planthopper (SBPH) Laodelphax striatellus and showed that six L. striatellus GluCl variants (LsGluCl-AS, LsGluCl-BS, LsGluCl-CS, LsGluCl-AL, LsGluCl-BL, LsGluCl-CL) were present in the SBPH. The expression patterns of six variants differed among developmental stages (egg, first- to fifth-instar nymphs, male and female adults) and among the body parts (head, thorax, abdomen, leg) of the female adult SBPH. All the transcripts were abundant in the head of the adult. When expressed in African clawed frog, Xenopus laevis, oocytes, the two functional variants (LsGluCl-AS, LsGluCl-AL) had similar EC50 and IC50 values for L-glutamate and channel blockers picrotoxinin and fipronil. CONCLUSION: This study represents a comprehensive molecular, expression and pharmacological characterisation of GluCl in the SBPH. These findings should be useful in providing more opportunities to discover novel insect control chemicals. © 2016 Society of Chemical Industry.

Impact evaluation of CONSORT and STRICTA guidelines on reporting quality for randomized controlled trials of acupuncture conducted in China.

OBJECTIVE: To evaluate and compare the reports' qualities of acupuncture randomized controlled trials (RCTs) conducted in China before and after the implementation of two guidelines, i.e., the Consolidated Standards of Reporting Trials (CONSORT) statement and the Standards for Reporting Interventions in Controlled Trials of Acupuncture (STRICTA). METHODS: Proportions of studies that reported CONSORT and STRICTA items were compared for the years before and after implementation of these two guidelines. In addition, the total score of each item's reporting were calculated and reported differences during different date ranges were compared. RESULTS: For CONSORT items (maximum score 8), there was evidence of a slight improvement in reporting between 1994-1995 and 1999-2000 combined (2.5±0.6) and 2004-2005 and 2009-2010 combined (3.0±0.9; difference 0.4, 95% confidence interval, 0.3 to 0.6, P<0.01). For STRICTA items (maximum score 17), there was evidence that a slight improvement in reporting between 1994-1995 and 1999-2000 combined (8.6±2.1) and 2004-2005 and 2009-2010 combined (10.1±1.8; difference 1.5, 95% confidence interval, 1.1 to 1.9, P<0.01). CONCLUSION: Quality of reporting for RCTs of acupuncture treatment conducted in China have generally improved since the introduction of the STRICTA and CONSORT guidelines.

Impact evaluation of CONSORT and STRICTA guidelines on reporting quality for randomized controlled trials of acupuncture conducted in China / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate and compare the reports' qualities of acupuncture randomized controlled trials (RCTs) conducted in China before and after the implementation of two guidelines, i.e., the Consolidated Standards of Reporting Trials (CONSORT) statement and the Standards for Reporting Interventions in Controlled Trials of Acupuncture (STRICTA).</p><p><b>METHODS</b>Proportions of studies that reported CONSORT and STRICTA items were compared for the years before and after implementation of these two guidelines. In addition, the total score of each item's reporting were calculated and reported differences during different date ranges were compared.</p><p><b>RESULTS</b>For CONSORT items (maximum score 8), there was evidence of a slight improvement in reporting between 1994-1995 and 1999-2000 combined (2.5±0.6) and 2004-2005 and 2009-2010 combined (3.0±0.9; difference 0.4, 95% confidence interval, 0.3 to 0.6, P<0.01). For STRICTA items (maximum score 17), there was evidence that a slight improvement in reporting between 1994-1995 and 1999-2000 combined (8.6±2.1) and 2004-2005 and 2009-2010 combined (10.1±1.8; difference 1.5, 95% confidence interval, 1.1 to 1.9, P<0.01).</p><p><b>CONCLUSION</b>Quality of reporting for RCTs of acupuncture treatment conducted in China have generally improved since the introduction of the STRICTA and CONSORT guidelines.</p>

[Post-stroke shoulder-hand syndrome treated with floating-needle therapy combined with rehabilitation training: a randomized controlled trial].

OBJECTIVE: To compare the clinical efficacy in the treatment of post-stroke shoulder-hand syndrome between floating-needle therapy and conventional acupuncture on the basis of rehabilitation training. METHODS: One hundred cases of post-stroke shoulder-hand syndrome were randomized into a floating-needle group and an acupuncture group, 50 cases in each one. The passive and positive rehabilitation training was adopted in the two groups. Additionally, in the floating-needle group, the floating-needle therapy was used. The needle was inserted at the site 5 to 10 cm away from myofasical trigger point (MTrP), manipulated and scattered subcutaneously, for 2 min continuously. In the acupuncture group, the conventional acupuncture was applied at Jianqian (EX-UE), Jianyu (LI 15), Jianliao (TE 14), etc. The treatment was given once every two days, 3 times a week, and 14 days of treatment were required. The shoulder hand syndrome scale (SHSS), the short form McGill pain scale (SF-MPQ) and the modified Fugl-Meyer motor function scale (FMA) were used to evaluate the damage severity, pain and motor function of the upper limbs before and after treatment in the two groups. The clinical efficacy was compared between the two groups. RESULTS: SHSS score, SF-MPQ score and FMA score were improved significantly after treatment in the two groups (all P < 0.01), and the improvements in the floating-needle group were superior to those in the acupuncture group (all P < 0.05). The total effective rate was 94.0% (47/50) in the floating-needle group, which was better than 90.0% (45/50) in the acupuncture group (P < 0.05). CONCLUSION: The floating-needle therapy combined with rehabilitation training achieves a satisfactory efficacy on post-stroke shoulder-hand syndrome, which is better than the combined therapy of conventional acupuncture and rehabilitation training.

Post-stroke shoulder-hand syndrome treated with floating-needle therapy combined with rehabilitation training: a randomized controlled trial / 中国针灸

<p><b>OBJECTIVE</b>To compare the clinical efficacy in the treatment of post-stroke shoulder-hand syndrome between floating-needle therapy and conventional acupuncture on the basis of rehabilitation training.</p><p><b>METHODS</b>One hundred cases of post-stroke shoulder-hand syndrome were randomized into a floating-needle group and an acupuncture group, 50 cases in each one. The passive and positive rehabilitation training was adopted in the two groups. Additionally, in the floating-needle group, the floating-needle therapy was used. The needle was inserted at the site 5 to 10 cm away from myofasical trigger point (MTrP), manipulated and scattered subcutaneously, for 2 min continuously. In the acupuncture group, the conventional acupuncture was applied at Jianqian (EX-UE), Jianyu (LI 15), Jianliao (TE 14), etc. The treatment was given once every two days, 3 times a week, and 14 days of treatment were required. The shoulder hand syndrome scale (SHSS), the short form McGill pain scale (SF-MPQ) and the modified Fugl-Meyer motor function scale (FMA) were used to evaluate the damage severity, pain and motor function of the upper limbs before and after treatment in the two groups. The clinical efficacy was compared between the two groups.</p><p><b>RESULTS</b>SHSS score, SF-MPQ score and FMA score were improved significantly after treatment in the two groups (all P < 0.01), and the improvements in the floating-needle group were superior to those in the acupuncture group (all P < 0.05). The total effective rate was 94.0% (47/50) in the floating-needle group, which was better than 90.0% (45/50) in the acupuncture group (P < 0.05).</p><p><b>CONCLUSION</b>The floating-needle therapy combined with rehabilitation training achieves a satisfactory efficacy on post-stroke shoulder-hand syndrome, which is better than the combined therapy of conventional acupuncture and rehabilitation training.</p>