[Transcutaneous electrical acupoint stimulation based on electro-oculogram signal regulation for children with mental retardation: a randomized controlled trial].

OBJECTIVE: To observe the clinical efficacy of transcutaneous electrical acupoint stimulation (TEAS) at Changqiang (GV 1) based on the modulation of electro-oculogram (EOG) signal for children with mental retardation, and explore the evaluation effect of the goal attainment scale (GAS) in children with mental retardation. METHODS: Sixty children with mental retardation were randomly divided into a treatment group and a control group, with 30 cases in each one. The children in the control group were treated with conventional rehabilitation, 5 times a week. On the basis of the control group, TEAS at Changqiang (GV 1) under the modulation of EOG signal was adopted in the treatment group. When the similarity between the collected EOG signal and the template was within the range of EOG threshold, one electric stimulation was triggered at Changqiang (GV 1) for 20 s (continuous wave, 70-100 Hz in frequency, 0.1-0.2 ms in pulse width), lasting 30 min in each treatment, the intervention was given twice a week. One course of treatment was composed of 4 weeks, and 3 courses were required in total in the two groups. The infant-junior high school student's social living ability scale (S-M) and GAS were scored and compared before and after treatment in the two groups. RESULTS: After treatment, the scores of self-living ability in the treatment group and communication ability in the control group were higher than those before treatment (P<0.01, P<0.05). The scores of collective activity and motor ability in the treatment group were higher than those in the control group (P<0.05). After treatment, GAS scores were higher than before treatment in both groups (P<0.001), and the score in the treatment group was higher than the control group (P<0.05). CONCLUSION: TEAS under the modulation of EOG signal is conductive to improving the collective, motor and self-living abilities of the children with mental retardation and promoting children's individual goals. Compared with the standard score of S-M, the T value of GAS can better reflect the subtle progress of individual.

Using Nonlinear Dynamics and Multivariate Statistics to Analyze EEG Signals of Insomniacs with the Intervention of Superficial Acupuncture.

OBJECTIVE: As a noninvasive and nonpharmacological therapeutic approach, superficial acupuncture (SA) is a special method of acupuncture. In this study, using nonlinear dynamics and multivariate statistics, we studied the electroencephalography (EEG) of primary insomnia under SA intervention to investigate how brain regions change. METHOD: This study included 30 adults with primary insomnia. They underwent superficial acupuncture at the Shangen acupoint. The EEG signals were collected for 10 minutes at each state, including the resting state, the intervention state, and the postintervention state. The data were conducted using nonlinear dynamics (including approximate entropy (ApEn) and correlation dimension (CD)) and multivariate statistics. RESULT: The repeated-measures ANOVA results showed that both ApEn and CD values were not significantly different at the three states (p > 0.05). The paired t-test results showed that the ApEn values of electrodes O2 (the right occipital lobe) at the postintervention state have decreased, compared with the resting state (p < 0.05), and no difference was detected in CD (p > 0.05). The cluster analysis results of ApEn showed that patients' EEG has changed from the right prefrontal lobe (electrode Fp2) to the right posterior temporal lobe (electrode T6) and finally to the right occipital lobe (electrode O2), before, during, and after the SA intervention. In addition, the factor analysis results of CD revealed that patients' EEG of all brain regions except for the occipital lobes has changed to the frontal lobes and anterior temporal and frontal lobes from pre- to postintervention. CONCLUSION: SA activated the corresponding brain regions and reduced the complexity of the brain involved. It is feasible to use nonlinear dynamics analysis and multivariate statistics to examine the effects of SA on the human brain.

[Electroacupuncture at "Jiaji" (EX-B2) induced down-regulation of OX-42 and P2X4 in lumbar spinal cord contributes to its analgesic after-effect in rats with chronic constriction injury].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) at "Jiaji" (EX-B2) at different time points on the expression of OX-42 (a monoclonal antibody with specific expression of complement receptor-3 in spinal microglial cells) and purinergic receptor P2X4 (P2X4) in rats with chronic constriction injury (CCI) of the sciatic nerve, as well as the possible after-effect mechanism of EA analgesia in neuropathic pain. METHODS: Sprague-Dawley rats were randomly divided into blank group, model group, immediately after EA group, 0.5-hour after EA group, 1-hour after EA group, 2-hour after EA group, 4-hour after EA group, 12-hour after EA group, and 24-hour after EA group, with 6 rats in each group. The rats in the model group and the EA groups were used to establish a model of CCI-induced neuropathic pain, and those in the immediately after EA group and the 0.5, 1, 2, 4, 12 and 24 hours after EA groups were treated with EA at bilateral L3 and L5 "Jiaji" points for 20 min after 7 d of modeling. Samples were collected immediately and at 0.5, 1, 2, 4, 12, and 24 hours after EA, and for the rats in the blank group and the model group, samples were collected after fixation the rats for 20 min. Heat pain threshold was observed before and after intervention, and immunohistochemistry was used to measure the protein expression of OX-42 and P2X4 in the spinal cord lumbar enlargement. RESULTS: After 7 days of modeling (before intervention), compared with the blank group, the heat pain threshold had a significant reduction in the model group and the EA groups (P<0.01). Compared with the model group after intervention, the immediately after EA group and the 0.5, 1 and 2 hours after EA groups had a significant increase in heat pain threshold (P<0.05). Compared with the model group, immediately after EA, the 0.5, 1 and 2 hours after EA groups had a significant reduction in the protein expression of OX-42 (P<0.01), and immediately after EA, the 0.5 and 1 hour after EA groups had a significant reduction in the protein expression of P2X4 (P<0.01). CONCLUSION: EA at "Jiaji" points can significantly increase heat pain threshold and down-regulate the protein expression of OX-42 and P2X4 in the spinal cord of CCI rats. The analgesic effect can last for 2 h.

[Multivariate statistical analysis of Evans blue exudation spots and acupoint sensitization distribution in rats with acute intestinal mucosal injury].

OBJECTIVE: To observe the distribution characters of Evans Blue (EB) exudation spots in the abdominal area in acute intestinal mucosal injury (AIMI) rats by using latitude and longitude grid counting and multivariate statistical analysis and to explore the feasibility of these methods. METHODS: Twenty-four SD rats were randomly divided into blank control, 7.5%, 10%, and 12.5% mustard oil groups (n＝6 rats in each group). The AIMI model was established by mustard oil enema, followed by injection of EB (0.1 mL/100g) into the tail vein. At 5, 10, 15, 20 and 25 h after EB injection, the rats under anesthesia were fixed in supine position for observing and photographing the abdominal subcutaneous EB exudation spots. The H．E. staining was used to observe histopathological changes of colonic mucosa. The longitude and latitude grids of the abdominal region were constructed (by taking the midpoint of the superior sternum as the origin) to determine the position of the blue spots. That the coordinate grids of the two regions can be connected geometrically is termed as "characteristic region". The data were processed by using multivariate statistical analysis. RESULTS: ① H．E. staining showed edema and inflammatory cell infiltration after colonic enema of different concentrations of mustard oil. ② Clustering analysis indicated that the distribution of exudation points in the "characteristic grid" had no temporal variation trend, and was not related with the concentration of mustard oil (P>0.05). ③ Factor analysis and contour analysis about the exudation spots of EB at 7.5% concentration showed that the "characteristic region Ⅱ" of different factors presented a tendency of time-dependent exudation, i．e. reduction of exudation degree along with time (P<0.05), and it is located near "Tianshu" acupoint. ④At 5 h after injection of EB, the 8 "characteristic regions" presented an EB-concentration-dependent tendency (reduction in exudation degree along with the increase of EB-concentration), among which the exudation degree of region C (near "Tianshu" acupoint) of the abdomen was higher (P<0.05). CONCLUSION: The multivariate statistical analysis method can be used to identify the abdominal "characteristic regions" of exudation spots of EB in rats with AIMI, and the characteristic region has acupoint sensitization characteristics related to the time and severity of mucosal injury.

[Metabonomics of Syndrome Markers in Infantile Bronchial Asthma Episode].

Objective To analyze urinary metabolites of bronchial asthma children patients with phlegm-heat obstructing Fei syndrome (PHOFS) and non-PHOFS using gas chromatography-mass spec- trometry/mass spectrometer ( GC-MS/MS) , thus performing research on syndrome markers. Methods Totally 44 bronchial asthma children patients with PHOFS and non-PHOFS in onset of asthma were recrui- ted. Another 29 healthy children were also recruited. Their urine samples were analyzed by GC-MS/MS. The profiles were analyzed using orthogonal partial least squares-discriminant analysis (OPLS-DA) , vari- able importance in the projection (VIP) , and non-parametric test to determine intergroup differential metabolites. Abnormal metabolic pathways were determined by Metaboanalyst. Results Compared with the health control group, contents of fourteen substances like inositol, uric acid, stearic acid, and so on de- creased, and mino-malonic acid content increased in asthma episode children (P <0. 05). The process was mainly involved in 5 metabolic pathways such as lysine degradation and biosynthesis, pyruvate me- tabolism, and so on. Compared with the non-PHOFS group in bronchial asthma episode, contents of nine substances like oxalic acid, L-threonine, pyrimidine, and so on decreased in the PHOFS group (P < 0. 05). The process was mainly involved in 5 metabolic pathways such as pentose phosphate pathway, inositol phosphate metabolism, and so on. Conclusions Urinary metabolites are different in infantile bronchial asthma episode and healthy children. Metabolic biomarkers and pathways exist in different syn- dromes in bronchial asthma episode.

Application of Traditional Chinese Medical Herbs in Prevention and Treatment of Respiratory Syncytial Virus.

Respiratory syncytial virus (RSV) is a common viral pathogen of the lower respiratory tract, which, in the absence of effective management, causes millions of cases of severe illness per year. Many of these infections develop into fatal pneumonia. In a review of English and Chinese medical literature, recent traditional Chinese medical herb- (TCMH-) based progress in the area of prevention and treatment was identified, and the potential anti-RSV compounds, herbs, and formulas were explored. Traditional Chinese medical herbs have a positive effect on inhibiting viral attachment, inhibiting viral internalization, syncytial formation, alleviation of airway inflammation, and stimulation of interferon secretion and immune system; however, the anti-RSV mechanisms of TCMHs are complicated, which should be further investigated.

Manual acupuncture at the SJ5 (Waiguan) acupoint shows neuroprotective effects by regulating expression of the anti-apoptotic gene Bcl-2.

Acupuncture at the SJ5 (Waiguan) acupoint has neuroprotective effects in cerebral infarction, but the underlying mechanism remains poorly understood. Here, we analyzed gene expression in healthy rat cerebellum using a pathway-focused DNA microarray to screen 113 genes associated with 18 signal transduction pathways. After 20 minutes of acupuncture at SJ5, the expression of Bcl-2 and Birc1b mRNA was markedly increased. This was confirmed by real-time reverse transcription PCR. Furthermore, western blot analysis showed that Bcl-2 protein expression remained high in the cerebellum until at least 2 hours after cessation of acupuncture. These findings indicate that acupuncture at SJ5 exerts neuroprotective effects by regulating the expression of anti-apoptotic gene Bcl-2.

Effects of Pinellia ternata (Thunb.) Berit. on the metabolomic profiles of placenta and amniotic fluid in pregnant rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Banxia (BX) is the root of Pinellia ternata (Thunb.) Berit. Its processed products, such as Jiang Banxia (JBX), have been clinically used in traditional Chinese medicine to treat vomiting, coughing, and inflammation. However, data for their safety for pregnant women are contradictory and confusing. AIM OF THE STUDY: To further explore the safety of BX, an ultra-performance liquid chromatography coupled with liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) metabolomics approach was used to evaluate the metabolic perturbation in pregnant rats caused by BX and JBX. MATERIALS AND METHODS: Placenta and amniotic fluid samples were collected from control Sprague-Dawley pregnant rats and exposed to BX suspension and JBX decoction (1.434g/kg/day). Samples were analyzed using LC-MS and GC-MS. The acquired MS data of above samples were further subjected to multivariate data analysis, and the significantly altered metabolites were identified. The associated pathways were constructed using MetaboAnalyst 3.0. RESULTS: The weight and histopathology of the placenta from each group of rats had no definite difference. However, we found 20 differential endogenous metabolites that changed significantly in the placenta and amniotic fluid samples. The alterations of identified metabolites indicated a perturbation in glycerophospholipid metabolism, amino acid metabolism, and carbohydrate metabolism in pregnant rats exposed to BX and JBX. CONCLUSION: In summary, this work suggested that oral administration of BX and JBX may induce disturbances in the intermediary metabolism in pregnant rats. This work contributes to further understanding the safety of BX and its processed products.

[Metabolomics analysis of Tripterygium wilfordii formulation based on theory of detoxicity compatibility].

Tripterygium wilfordii Hook. f. induced-hepatotoxicity was the main limitation for its usage in clinic. Qingluo Tongbi formulation showed obvious attenuation for hepatotoxicity in clinic and fundamental research in vivo. To explore the potential mechanism of the attenuation, we conducted a study on the plasma metabolomic profiles of T. wilfordii and Qingluo Tongbi formulation in rats by a sensitive gas chromatography-mass spectrometry (GC-MS/MS) method. In plasma samples, a total of 72 compounds were analyzed by EI source MS, and were successfully identified by matching NIST database. The semi-quantification results were then calculated by OPLS-DA model with SIMCA-P 13.0 software. The three groups were clearly distinguished in OPLS-DA score plot. In addition, the observation values of Qingluo Tongbi formulation showed the obvious trend towards the control levels, suggesting the detoxicity effect of the formulation. Variation metabolites were further analyzed by VIP and One Way ANOVAs, and the results showed a significant increase in compounds of glycogenic amino acids, such as alanine, proline, serine and glutamine after the administration of T. wilfordii, indicated that the tissue proteins were decomposed and amino acids were leakage into blood. Qingluo Tongbi formulation could reverse the amino acids into normal level. On the contrary, the levels of glucose, lactic acid and hydroxy butyrate decrease, and the formulation can relieve the disorder in the levels of lactic acid, suggesting the regulation of the energy metabolism. Additionally, the level of branched chain amino acid was decreased, suggested the toxicity was induced, but the formulation cannot increase it into the normal levels. Nevertheless, all the above results suggested that the classical Qingluo Tongbi formulation displayed the liver protection effect by adjusting the amino acid levels and regulating the energy metabolism. Qingluo Tongbi formulation was developed based on traditional Chinese medicine theory "detoxicity compatibility", and contained Panax notoginseng (Burk.) F. H. Chen to nourish blood and absorb clots. Modern pharmacology suggested that its liver protection effect was correlated with the promotion of protein synthesis. Another important herb is Rehmannia glutinosa Libosch., which can regulate the energy metabolism. Both were consistent with the metabolomic results in this study, which explained the potential mechanism of "detoxicity compatibility" theory. Therefore, the currently developed metabolomic approach and the obtained results would be highly useful for the comprehensive toxicity studies for other herbal medicines and various complex deoxicity formulations.

A combination of neostigmine and anisodamine protects against ischemic stroke by activating α7nAChR.

BACKGROUND: Increasing endogenous acetylcholine by neostigmine decreased the ischemic cerebral injury. The off-target action on muscarinic receptor produced a variety of adverse effects and limited the clinical application on stroke. AIM: We combined neostigmine with anisodamine and investigated the neuroprotection and mechanism. METHODS: Male Sprague-Dawley rats were subjected to middle cerebral artery occlusion. Neuroprotective action of neostigmine in combination with anisodamine at varying ratios was examined to determine the optimal combination as well as ideal therapeutic window. Potential involvement of α7 nicotinic acetylcholine receptor was examined by measuring the infarct size, the expression of proinflammatory cytokines, and the biomarkers of apoptosis in α7 nicotinic acetylcholine receptor knockout mice. A set of in vitro experiments was conducted in RAW264.7 cells to probe into potential molecular mechanisms. RESULTS: The neostigmine/anisodamine combination conferred neuroprotection. The protection was most potent at a ratio of 1:500. At such a ratio, the combination increased the binding of acetylcholine to α7 nicotinic acetylcholine receptor and reduced proinflammatory cytokines. The neuroprotection was evident only in wild-type and not in α7 nicotinic acetylcholine receptor knockout mice. The combination significantly decreased the expression of Bad and Bax, and increased Bcl-2 and Bcl-xl in α7 nicotinic acetylcholine receptor wild-type mice but not in knockout mice. The combination did not affect caspase-8, cleaved caspase-8, or caspase-12. CONCLUSIONS: Current study identified the optimal combination of neostigmine and anisodamine against ischemic stroke, and indicated that the acetylcholine-α7 nicotinic acetylcholine receptor is involved in the protective effects.

Epimedium flavonoids ameliorate experimental autoimmune encephalomyelitis in rats by modulating neuroinflammatory and neurotrophic responses.

The present study was designed to determine whether epimedium flavonoids (EF) had effect on the development of experimental autoimmune encephalomyelitis (EAE) in rats and to elucidate its underlying mechanisms. EAE was induced by immunization of adult female Lewis rats with partially purified myelin basic protein (MBP) prepared from guinea-pig spinal cord homogenate. EF was administrated intragastrically once a day after immunization until day 14 post immunization (p.i.). Histopathological staining, enzyme-linked immunosorbent assay (ELISA), biochemical methods and western blotting approaches were used to evaluate the disease incidence and severity, neuroinflammatory and neurotrophic response in the central nervous system (CNS). Intragastrical administration of EF (20 and 60 mg/kg) significantly reduced clinical score of neurological deficit in EAE rats; alleviated demyelination and inflammatory infiltration; and inhibited astrocytes activation, production of proinflammatory molecules such as interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α), nitric oxide (NO) and nuclear transcription factor (NF-κB) in the spinal cord of EAE rats. Treatment with EF also enhanced the expression of 2', 3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) and nerve growth factor (NGF), increased the number of oligodendrocytes and protected the ultrastructure of myelin sheaths and axons in the spinal cord of EAE rats. Our results showed that EF inhibited the development of partial MBP-induced EAE in rats. This effect involved reducing neuroinflammation and enhancing myelination and neurotrophins and our findings suggest that EF may be useful for the treatment of multiple sclerosis.

Feeding deterrents from Aconitum episcopale roots against the red flour beetle, Tribolium castaneum.

The screening for insecticidal principles from several Chinese medicinal herbs showed that the ethanol extract of Aconitum episcopale roots possessed significant feeding deterrence against the red flour beetle, Tribolium castaneum . From the ethanol extract, six feeding deterrents were isolated by bioassay-guided fractionation. The compounds were identified as chasmanine, crassicauline A, karacoline, sachaconitine, talatisamine, and yunaconitine from their spectroscopic data. Chasmanine, talatisamine, karacoline, and sachaconitine exhibited feeding deterrent activity against T. castaneum adults, with EC(50) values of 297.0, 342.8, 395.3, and 427.8 ppm, respectively. Yunaconitine and crassicauline A also possessed feeding deterrent activity against T. castaneum adults, with EC(50) values of 653.4 and 1134.5 ppm, respectively.

[Clinical observation on treatment of primary dysmenorrhea with acupuncture and massage].

OBJECTIVE: To observe the clinical therapeutic effect of acupuncture massage for primary dysmenorrhea and its influence on prostaglandin F2alpha level in menstrual fluid. METHODS: Ninety patients with primary dysmenorrhea were assigned equally to the acupuncture group, the massage group and the control group (oral administrated with Somiton tablet). The treatment course was 3 menstrual cycles. The effect in relieving menalgia was evaluated using numerical rating scale (NRS) before and after treatment and change of PGF2alpha content in menstrual fluid was tested as well. RESULTS: The NRS score was 0.60 +/- 0.67 in the acupuncture group and 0.53 +/- 0.63 in the massage group respectively, significantly lower than that in the control group, which was 2.30 +/- 1.73 (P < 0.01). The PGF2alpha level after treatment was significantly lower in the acupuncture group (176.58 +/- 17.85 ng/L) and the massage group (171.32 +/- 17.15 ng/L) than that in the control group (220.56 +/- 54.47 ng/L), respectively (P <0.01). There showed no difference in the two aspects between the acupuncture group and the massage group (P >0.05). CONCLUSION: Acupuncture and massage has a reliable therapeutic effect on primary dysmenorrhea, which is superior to that of Somiton tablet, and its mechanism might be exerted by regulating PGF2alpha level in menstrual fluid.