Neurotoxicity study of ethyl acetate extract of Zanthoxylum armatum DC. on SH-SY5Y based on ROS mediated mitochondrial apoptosis pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Zanthoxylum armatum DC. (ZADC) is a traditional medicinal plant with various pharmacological activities and is widely used in China, Japan, India, and other regions. Previous studies have revealed that the methanol extract of ZADC can cause neurotoxicity symptoms in rats, such as drooling, decreased appetite, decreased movement, and increased respiratory rate. However, the basis of these toxic substances and the mechanism of neurotoxicity remain unclear. AIM OF THE STUDY: To evaluate the effects of ZADC on nerve cells and their damage mechanisms and discuss the possible toxic substance basis. MATERIALS AND METHODS: The ethyl acetate extract of ZADC is obtained by extracting the methanol extract of ZADC with ethyl acetate. The Q-Orbitrap LC-MS/MS method was employed to analyze the chemical composition of the EA extract of ZADC. SH-SY5Y cells were incubated with different concentrations of the ethyl acetate extract of ZADC. The cytotoxicity of the extract was evaluated using CCK-8, LDH, and ROS assays, and the oxidative stress status of cells was assessed using MDA, GSH, and SOD. Cell apoptosis was detected using flow cytometry. Damage to mitochondrial function was evaluated by labeling mitochondria, ATP, and MMP with fluorescence. Cyto-C, Caspase-3, Caspase-9, Apaf-1, Bax, and reduced Bcl2 expression were measured to evaluate the activation of the mitochondrial apoptosis pathway. Finally, NAC intervention was used to detect changes in the relevant indicators. The activation of mitochondrial apoptosis pathway was evaluated by measuring Cyto-C, Caspase-3, Caspase-9, Apaf-1, and Bax and Bcl2 expression. Finally, NAC intervention was utilized to detect changes in the relevant indicators. RESULTS: After treating SY-SY5Y cells with EA extract from ZADC, cell viability decreased significantly, and the intracellular ROS level increased in a dose-dependent manner. Meanwhile, ZADC can cause cellular oxidative stress and increase MDA and SOD concentrations while decreasing GSH concentrations. It can also shorten the mitochondrial cristae and decrease the number of mitochondria. In contrast, it can reduce ATP synthesis in the mitochondria and mitochondrial membrane potential (MMP). Furthermore, it increased the apoptosis rate and the expression of Cyto-C, Caspase-3, Caspase-9, Apaf-1, and Bax and reduced Bcl2 expression. NAC intervention alleviated the reduction in SH-SY5Y cell survival and the accumulation of reactive oxygen species induced by the EA extract in ZADC. It also inhibits signaling pathways dominated by proteins, such as Cyto-C, reducing cell apoptosis and cytotoxicity. A total of 46 compounds were identified in the extracts. CONCLUSIONS: The results suggest that EA extract of ZADC can induce the mitochondrial apoptotic pathway by accumulating ROS in cells, leading to apoptosis. Antioxidants had a good inhibitory and protective effect against cell damage caused by the EA extract of ZADC. The neurotoxic components of ZADC may be organic acids and compounds containing amino groups.

Effect of Acupuncture Combined with Low Frequency Electrical Stimulation on Cerebral Palsy Salivation / 中国康复理论与实践

Objective To explore the effect of oral sensorimotor therapy (OSMT), acupuncture, and low frequency electrical stimulation on salivation in patients with cerebral palsy. Methods From September, 2014 to October, 2015, 80 cerebral palsy patients with salivation were recruited. They were randomly divided into four treatment groups:group A was treated with OSMT, group B was treated with acupunc-ture, group C was treated with frequency electrical stimulation, and group D was treated with acupuncture and low frequency electrical stim-ulation, for eight weeks. Meanwhile, 20 cerebral palsy patients with salivation from outpatients were chosen as control group. Results There was no improvement in the root mean square (RMS) value of surface electromyography (sEMG) after treatment in both the control group and group A (t0.05), and the RMS value significantly improved in the groups B, C and D (t>8.983, P0.05), and they was better in the groups B, C and D than in the control group and group A (W>226.0, t>8.534, P306.0, t>3.663, P<0.05). Conclusion Acupuncture and low frequency electrical stimulation could effectively improve salivation in children with cerebral palsy, and their combination was superior to either ones, while OSMT did not show obvious benefit.

Systematic Review on Vertigo Treatment with Tian-Ma Gou-Teng Decoction and Its Modifications / 世界科学技术-中医药现代化

This study was aimed to make a systematic and comprehensive evaluation on clinical efficacy and safety of vertigo treatment with Tian-Ma Gou-Teng (TMGT) Decoction. Articles had been searched in the Chinese Journal Full-text Database, Chongqing VIP Chinese Scientific Journals Database, China National Knowledge Infrastructure (CNKI), Chinese Biomedical Literature Database Online, Wanfang Data and Chinese Medicine Journal Literature Database for randomized controlled trials (RCT) on vertigo treatment with TMGT Decoction. The data retrieval time was from the establishment time of the database until present. Two investigators screened literatures, extracted data and assessed quality according to inclusion and exclusion criteria independently. RevMan5.0.2 was used in the meta-analysis. The results showed that a total of 33 trials were involved, which covered 2922 vertigo cases. The meta-analysis showed that from the aspect of clinical effectiveness, the comparison between Chinese medicine treatment group and modern medicine treatment group (OR = 3.67, 95% CI [2.66~5.07]), comparison between integrative medicine treatment group and modern medicine treatment group (OR = 3.28, 95% CI [2.33~4.62]), the comparison between Chinese medicine combined with other method (such as acupuncture, Tuina) treatment group and other Chinese medicine treatment group (OR = 2.29, 95% CI [1.34~3.91]), therapeutic effect in the experimental group was better than the control group. For the aspect of relapse rate, TMGT Decoction in the treatment of vertigo was better than the control group (OR = 0.27, 95% CI [0.12~0.65]). In the aspect of adverse reactions, TMGT Decoction in the treatment of vertigo was better than the control group (OR = 0.40, 95% CI [0.13~1.24]). For aspects of other indicators such as quality of life, no further analysis was conducted since there was no complete reported literature. It was concluded that the system evaluation results showed that compared with other methods, TMGT Decoction has better clinical efficacy and low recurrence rate, but safety needs to be further eualuated. However, due to the quality of included studies was not high, current evidences cannot be determined to be fully applicable in the clinical practice. But its efficacy advantages are worthy of further research. It especially requires high quality multi-center, large sample, double-blind, RCTs to verify.