Acupuncture at Back-Shu point improves insomnia by reducing inflammation and inhibiting the ERK/NF-κB signaling pathway.

BACKGROUND: Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep. Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia, and long-term use is prone to drug resistance and other adverse reactions. Acupuncture has a good curative effect and unique advantages in the treatment of insomnia. AIM: To explore the molecular mechanism of acupuncture at Back-Shu point for the treatment of insomnia. METHODS: We first prepared a rat model of insomnia, and then carried out acupuncture for 7 consecutive days. After treatment, the sleep time and general behavior of the rats were determined. The Morris water maze test was used to assess the learning ability and spatial memory ability of the rats. The expression levels of inflammatory cytokines in serum and the hippocampus were detected by ELISA. qRT-PCR was used to detect the mRNA expression changes in the ERK/NF-κB signaling pathway. Western blot and immunohistochemistry were carried out to evaluate the protein expression levels of RAF-1, MEK-2, ERK1/2 and NF-κB. RESULTS: Acupuncture can prolong sleep duration, and improve mental state, activity, diet volume, learning ability and spatial memory. In addition, acupuncture increased the release of 1L-1ß, 1L-6 and TNF-α in serum and the hippocampus and inhibited the mRNA and protein expression of the ERK/NF-κB signaling pathway. CONCLUSION: These findings suggest that acupuncture at Back-Shu point can inhibit the ERK/NF-κB signaling pathway and treat insomnia by increasing the release of inflammatory cytokines in the hippo-campus.

Different proteomic profiles of cinnabar upon therapeutic and toxic exposure reveal distinctive biological manifestations.

ETHNOPHARMACOLOGICAL RELEVANCE: Cinnabar, a traditional Chinese mineral medicine with sedative and tranquilizing effects, is known to be toxic to the neural system, but its detailed pharmacological and toxicological mechanisms are still unclear. AIM OF THE STUDY: This study aimed to explore the potential neuropharmacological and neurotoxicological mechanisms of cinnabar by investigating the differentially expressed proteins in cerebral cortices of mice exposed to therapeutic and toxic doses of cinnabar. MATERIALS AND METHODS: Label-free quantitative proteomics and bioinformatics analysis were used to characterize the proteins, pathways, and potential targets associated with therapeutic (50 mg/kg) and toxic (1000 mg/kg) doses of cinnabar in cerebral cortices of mice. Proteomic analysis was verified by parallel reaction monitoring. RESULTS: A total of 6370 and 6299 proteins were identified in the cerebral cortices of mice after exposure to therapeutic and toxic doses of cinnabar, among which 130 and 119 proteins were differentially expressed, respectively. Functional/pathway enrichment analysis showed that both exposure doses of cinnabar could affect transport processes in the cerebral cortex through different proteins. The changes induced by the therapeutic dose included pathways involved in translation and sphingolipid metabolism. Interestingly, for the toxic dose, differentially expressed proteins were enriched for functions and pathways related to RNA splicing, transcription, synaptic plasticity regulation and developmental processes, among which RNA splicing was the most significantly affected function. ATP6V1D and CX3CL1 were shown to be possible key proteins affected by cinnabar, leading to multiple functional changes in the cerebral cortex at the therapeutic and toxic doses, respectively. Furthermore, Connectivity Map (CMap) analysis predicted LRRK2 to be a potential therapeutic target and FTase to be a potential toxic target for cinnabar. CONCLUSION: Our results suggest that the pathways and potential targets identified in the mouse cerebral cortex exposed to therapeutic and toxic doses of cinnabar are different, which provides novel insights into the potential molecular mechanisms underlying the pharmacological and toxicological effects of cinnabar.

Effectiveness and safety of acupuncture for insomnia: Protocol for a systematic review.

BACKGROUND: Insomnia is one of the most common diseases in modern society, the main characteristics of the patients were difficulty in falling asleep at night and/or failure to maintain effective sleep after falling asleep. It can lead to early awakening, short sleep, heavy sleeplessness, dreaming, poor sleep quality, and working hours after waking up, causes a series of negative emotions, such as fatigue, inefficiency, cognitive decline, social interaction, tension, and anxiety, which affect social harmony and stability. So Insomnia has gained more and more attention. At present, acupuncture has been proved effective in the treatment of insomnia by many studies. The purpose of this study is to evaluate the efficacy and safety of acupuncture in the treatment of insomnia, and to provide the latest evidence for clinical application. METHODS AND ANALYSIS: We collected the qualified literature on acupuncture treatment of insomnia by electronic retrieval of Cochrane Library, China National Knowledge Infrastructure (CNKI), China Biomedical Disc (CBMDISC), PubMed, China Science and Technology Journal Database (VIP) and Wanfang Database, and manual retrieval of papers and internal reports. We will select the eligible studies published up to September 30, 2019. We use Insomnia Severity Index (ISI) as the main outcome of insomnia and Pittsburgh Sleep Quality Index (PSQI), Hamilton Depression Scale(HAMD) and Self-Rating Anxiety Scale (SAS) as secondary indicators to evaluate the efficacy and safety of acupuncture treatment of insomnia, we will use Revman v.5.3 software to calculate data synthesis, and if the results are appropriate, meta-analysis can also be carried out. RESULTS: This study will provide comprehensive evidence of high quality of acupuncture treatment for insomnia from ISI, PSQI, HAMD, SAS, and adverse reactions. CONCLUSION: The systematic review will provide a basis for evaluating the efficacy and safety of acupuncture in the treatment of insomnia. TRIAL REGISTRATION NUMBER: PROSPERO CRD42019131957.

Effect of Euphorbia factor L1 on intestinal barrier impairment and defecation dysfunction in Caenorhabditis elegans.

BACKGROUND: Euphorbia factor L1 (EFL1) is a lathyrane-type diterpenoid from the medicinal herb Euphorbia lathyris L., and has been reported with intestinal toxicity, but the potential mechanisms remain unknown. PURPOSE: The objective of this study was to investigate the intestinal toxicity of EFL1 and the underlying mechanisms using nematode Caenorhabditis elegans. METHODS: C. elegans were exposed to 0-200 µM EFL1 for 72 h, then the survival rate, body length and body width, locomotion and chemoreception behavior, intestinal ROS and lipofuscin accumulation, intestinal permeability, and defecation rhythm were detected. The γ-aminobutyric acid（GABA) energic neurons AVL and DVB were shown via green fluorescent protein under a laser scanning confocal microscope. The structure of GABA transporter UNC-47 were predicted by homology modeling, and the interaction between EFL1 and UNC-47 was simulated by molecular docking. The mRNA expression of genes related to oxidative stress, intestinal permeability and defecation after EFL1 exposure were detected by RT-qPCR. RESULTS: EFL1 did not induce lethality of nematodes. The general toxicity was characterized by abnormal growth, locomotion and chemoreception. The intestinal barrier was leaky, due to down-regulated cell junction and active cation transport. The mean defecation cycle length in nematodes was decreased, relating to disorder vesicular and ion transport, enhanced rhythm behavior and muscle contraction. The dysfunctional defecation also attributed to injured UNC-47 protein, as well as GABAergic neurons AVL and DVB. Excessive ROS and lipofuscin accumulation were observed in intestine, along with activation of antioxidant enzymes of SOD, COQ7 and CAT. CONCLUSION: This study elucidated the EFL1-induced intestinal toxicity in nematodes, characterized as leaky intestinal barrier and accelerated defecation behavior. The underlying mechanisms were involved in oxidative stress, cell junctions, transportation, rhythm behavior, muscle contraction, and GABAergic neurons.

Effect of euphorbia factor L1 on oxidative stress, apoptosis, and autophagy in human gastric epithelial cells.

BACKGROUND: Euphorbia factor L1 (EFL1), a lathyrane-type diterpenoid from the medicinal herb Euphorbia lathyris L. (Euphorbiaceae), has been reported for many decades to induce gastric irritation, but the underlying mechanisms remain unclear. PURPOSE: The objective of this study was to investigate EFL1-induced cytotoxicity and the potential mechanisms of action on the human normal gastric epithelial cell GES-1. METHODS: GES-1 cells were treated with EFL1 (12.5-200 µM) for different time intervals, and cell survival, LDH release, intracellular reactive oxygen species (ROS), malondialdehyde (MDA) content, and superoxide dismutase (SOD) activity were detected. Mitochondrial membrane potential (MMP) assay, DAPI staining, DNA fragment assay, and annexin V-FITC/PI staining were performed. The interaction between EFL1 and Bcl-2, cytochrome c, caspase-9, caspase-3, PI3K, AKT, and mTOR proteins was simulated by molecular docking. The mRNA and protein expression of apoptosis and autophagy factors were detected by RT-qPCR and Western blotting. RESULTS: EFL1 decreased survival, increased LDH leakage, and induced abnormal production of ROS, MDA and SOD in GES-1 cells. Mitochondria-mediated apoptosis was characterized by decreased MMP, condensed nuclei, fragmented DNA, and increased apoptosis rate. EFL1 interacted with proteins via hydrogen bonding. The mRNA, total or phosphorylated protein expression of Bcl-2, mitochondrial cytochrome c, PI3K, AKT, mTOR and p62 were downregulated; in contrast, those of cytoplasmic cytochrome c, cleaved caspase-9, cleaved caspase-3, LC3-ll and Beclin-1 were upregulated. CONCLUSION: These findings indicated that EFL1 decreased the survival of GES-1 cells through EFL1-induced oxidative stress, activation of the mitochondria-mediated apoptosis as well as autophagy via inhibition of the PI3K/AKT/mTOR pathway.

Intraventricular administration of tigecycline for the treatment of multidrug-resistant bacterial meningitis after craniotomy: a case report.

BACKGROUND: Intracranial infections, especially multidrug-resistant (MDR) bacterial meningitis, are one of the most severe complications after craniotomy and may greatly impact patient outcomes. CASE PRESENTATION: We report a case of severe MDR Klebsiella pneumonia meningitis after craniotomy that was treated with three different dosages of tigecycline (Pfizer, New York, NY, U.S.A.)via a combined intravenous (IV) and intracerebroventricular (ICV) administration. Here, we discuss the pharmacokinetics (PK) of a combined IV and ICV tigecycline administration for a patient with an intracranial infection after craniotomy. CONCLUSION: In the present case, three different dosages of tigecycline were administered: 49 mg IV plus 1 mg ICV q12 h, 45 mg IV plus 5 mg ICV q12 h, 40 mg IV plus 10 mg ICV q12 h. The combined IV and ICV administration might improve CSF tigecycline concentrations, and in this case, the methods of administration were safe and effective.

Shikonin kills glioma cells through necroptosis mediated by RIP-1.

BACKGROUND AND PURPOSE: Shikonin was reported to induce necroptosis in leukemia cells, but apoptosis in glioma cell lines. Thus, it is needed to clarify whether shikonin could cause necroptosis in glioma cells and investigate its underlying mechanisms. METHODS: Shikonin and rat C6 glioma cell line and Human U87 glioma cell line were used in this study. The cellular viability was assayed by MTT. Flow cytometry with annexin V-FITC and PI double staining was used to analyze cellular death modes. Morphological alterations in C6 glioma cells treated with shikoinin were evaluated by electronic transmission microscopy and fluorescence microscopy with Hoechst 33342 and PI double staining. The level of reactive oxygen species was assessed by using redox-sensitive dye DCFH-DA. The expressional level of necroptosis associated protein RIP-1 was analyzed by western blotting. RESULTS: Shikonin induced cell death in C6 and U87 glioma cells in a dose and time dependent manner. The cell death in C6 and U87 glioma cells could be inhibited by necroptosis inhibitor necrotatin-1, not by pan-caspase inhibitor z-VAD-fmk. Shikonin treated C6 glioma cells presented electron-lucent cytoplasm, loss of plasma membrane integrity and intact nuclear membrane in morphology. The increased ROS level caused by shikonin was attenuated by necrostatin-1 and blocking ROS by anti-oxidant NAC rescued shikonin-induced cell death in both C6 and U87 glioma cells. Moreover, the expressional level of RIP-1 was up-regulated by shikonin in a dose and time dependent manner as well, but NAC suppressed RIP-1 expression. CONCLUSIONS: We demonstrated that the cell death caused by shikonin in C6 and U87 glioma cells was mainly via necroptosis. Moreover, not only RIP-1 pathway, but also oxidative stress participated in the activation of shikonin induced necroptosis.

[Sequential acupuncture method for treatment of apoplexy hemiplegia and its effects on plasma ET-1 and NO levels].

OBJECTIVE: To observe clinical therapeutic effect of sequential acupuncture method on apoplexy hemiplegia and to probe into the mechanism. METHODS: One hundred and forty-four cases of apoplexy hemiplegia were randomly divided into a treatment group treated with sequential acupuncture method and a control group treated with traditional acupuncture, 72 cases in each group. After treatment for 3 months, their clinical therapeutic effects were observed and plasma endothelin-1 (ET-1) and nitric oxide (NO) levels were detected. RESULTS: The treatment group in the improvement of the nerve function-deficiency, total life ability and clinical overall effect, and the course of recovery, decreasing of ET-1 level and increasing of NO level was superior to the control group (P < 0.05, P < 0.01), the shorter the course, the better the therapeutic effect. CONCLUSION: The sequential acupuncture method has an obvious therapeutic effect on apoplexy hemiplegia, and it can decrease plasma ET-1 level and increase NO level. It is an effective therapy for apoplexy hemiplegia.