[Interactive scalp acupuncture for hemiplegic upper extremity motor dysfunction in patients with ischemic stroke: a randomized controlled trial].

OBJECTIVE: To compare the curative effect between interactive scalp acupuncture and traditional scalp acupuncture on hemiplegic upper extremity motor dysfunction in the patients with ischemic stroke. METHODS: Seventy cases of hemiplegic upper extremity motor dysfunction of ischemic stroke were randomly divided into an interactive scalp acupuncture group (35 cases, 1 case breaked off) and a traditional scalp acupuncture group (35 cases, 1 case dropped off). The patients of the two groups received the secondary prevention medication and routine rehabilitation therapy. Besides, in the interactive scalp acupuncture group, the upper extremity occupational therapy was operated during the needle retaining of scalp acupuncture; and in the traditional scalp acupuncture group, the upper extremity occupational therapy was delivered after the completion of scalp acupuncture. The same points were selected in the two groups such as Fuxiang head area, Fuxiang upper-limb-shoulder point, Fuxiang upper-limb-elbow point and Fuxiang upper-limb-wrist point. The needles were inserted perpendicularly by flying-needle technique and manipulated by triple technique of gentle twisting, heavy pressure and vibrating. The needles were retained for 30 min. Based on the degree of the upper extremity motor impairment, the regimen of the upper extremity occupational therapy was formulated individually and one treatment took 30 min. In the two groups, the therapies were delivered once daily, 5 times a week, lasting 4 weeks. Before and after treatment, the scores of Fugl-Meyer assessment of upper extremity (FMA-UE), Wolf motor function test (WMFT), the modified Barthel index (MBI) and the modified Ashworth scale (MAS) grade in the two groups were observed before and after treatment. RESULTS: After treatment, the scores of FMA-UE, WMFT and MBI were higher than those before treatment (P<0.01), and MAS grade was improved (P<0.05) in the two groups. The scores of FMA-UE, WMFT and MBI in the interactive scalp acupuncture group were higher than those in the traditional scalp acupuncture group (P<0.01, P<0.05), and there was no statistical significance in the difference of MAS grade between the two groups (P>0.05). CONCLUSION: The interactive scalp acupuncture can effectively improve the motor function of the hemiplegic upper extremities and the activities of daily living in the patients with ischemic stroke and its efficacy is better than traditional scalp acupuncture. But these two types of scalp acupuncture obtain the similar effect on spasticity.

[Effect of Tongli Shuji acupuncture on the expression of Jagged2 and Notch2 in a rat model of permanent ischemia of the right middle cerebral artery].

OBJECTIVE: To investigate the effect of Tongli Shuji acupuncture on the behavioristics, the percentage of cerebral infarct volume, and the expression of Jagged2 and Notch2 proteins in the hippocampus of the ischemic hemisphere in rats with permanent ischemia of the right middle cerebral artery, as well as the mechanism of Tongli Shuji acupuncture in improving ischemic cerebrovascular disease. METHODS: Sprague-Dawley rats were randomly divided into sham-operation group, model control group, medication group, routine acupuncture group, and Tongli Shuji group, with 10 rats in each group. The suture method was used to establish a model of permanent ischemia of the right middle cerebral artery. The rats in the medication group were given Citicoline (0.4 mg/kg) by gavage, those in the routine acupuncture group were given acupuncture at"Baihui"(GV20) and"Dazhui" (GV14) 15 minutes per day, and those in the Tongli Shuji group were given acupuncture at "Zhengying "(GB17), "Tianjing"(SJ10), and"Huantiao"(GB30) at the left side. The neurological deficit score was determined on days 1, 7, 14, 21 and 28 after surgery, the percentage of cerebral infarct volume was measured after treatment, and Western blot was used to measure the protein expression of Jagged2 and Notch2 in the right hippocampal tissue. RESULTS: Compared with the sham-operation group, the model control group had significant increases in the neurological deficit score on day 1 after surgery (P<0.001). Compared with the model control, medication and routine acupuncture groups, the Tongli Shuji group had significant reductions in the neurological deficit score on day 21 after surgery (P<0.01). Compared with the sham-operation group, the model control group had a significant increase in the percentage of cerebral infarct volume(P<0.05)．The Tongli Shuji group had a significant reduction in the percentage of cerebral infarct volume compared with the medication groups (P<0.05). Compared with the model control group, the routine acupuncture group had a significant increase in the expression of hippocapmal Jagged2(P<0.01)．Compared with the model control, medication and routine acupuncture groups, the Tongli Shuji group had significant increases in the expression of hippocampal Jagged2 and Notch2 (P<0.01). CONCLUSION: Acupuncture can improve the behavioral manifestations of neurological deficit in rats with permanent ischemia of the right middle cerebral artery. Tongli Shuji acupuncture can significantly up-regulate the protein expression of Jagged2 and Notch2 in the hippocampal tissue of the ischemic region, possibly by activating the Notch signaling pathway to exert a neuroprotective effect.

Tetrahydroxystilbene glucoside relieves the chronic inflammatory pain by inhibiting neuronal apoptosis, microglia activation, and GluN2B overexpression in anterior cingulate cortex.

Tetrahydroxystilbene glucoside (THSG) is one of the active ingredients of Polygonum multiflorum. It has been shown to exert a variety of pharmacological effects, including antioxidant, anti-aging, and anti-atherosclerosis. Because of its prominent anti-inflammatory effect, we explored whether THSG had analgesic effect. In this study, we used a model of chronic inflammatory pain caused by injecting complete Freund's adjuvant into the hind paw of mice. We found THSG relieved swelling and pain in the hind paw of mice on a dose-dependent manner. In the anterior cingulate cortex, THSG suppressed the upregulation of GluN2B-containing N-methyl-D-aspartate receptors and the downregulation of GluN2A-containing N-methyl-D-aspartate receptors caused by chronic inflammation. In addition, THSG increased Bcl-2 and decreased Bax and Caspase-3 expression by protecting neuronal survival. Furthermore, THSG inhibited the phosphorylation of p38 and the increase of nuclear factor κB (NF-κB) and tumor necrosis factor α (TNF-α). Immunohistochemical staining revealed that THSG blocked the activation of microglia and reduced the release of proinflammatory cytokines TNF-α, interleukin 1ß (IL-1ß), and interleukin 6 (IL-6). In conclusion, this study demonstrated that THSG had a certain effect on alleviating complete Freund's adjuvant-induced chronic inflammatory pain.

Compound to Extract to Formulation: a knowledge-transmitting approach for metabolites identification of Gegen-Qinlian Decoction, a traditional Chinese medicine formula.

Herbal medicines usually contain a large group of chemical components, which may be transformed into more complex metabolites in vivo. In this study, we proposed a knowledge-transmitting strategy for metabolites identification of compound formulas. Gegen-Qinlian Decoction (GQD) is a classical formula in traditional Chinese medicine (TCM). It is widely used to treat diarrhea and diabetes in clinical practice. However, only tens of metabolites could be detected using conventional approaches. To comprehensively identify the metabolites of GQD, a "compound to extract to formulation" strategy was established in this study. The metabolic pathways of single representative constituents in GQD were studied, and the metabolic rules were transmitted to chemically similar compounds in herbal extracts. After screening diversified metabolites from herb extracts, the knowledge was summarized to identify the metabolites of GQD. Tandem mass spectrometry (MSn), fragment-based scan (NL, PRE), and selected reaction monitoring (SRM) were employed to identify, screen, and monitor the metabolites, respectively. Using this strategy, we detected 131 GQD metabolites (85 were newly generated) in rats biofluids. Among them, 112 metabolites could be detected when GQD was orally administered at a clinical dosage (12.5 g/kg). This strategy could be used for systematic metabolites identification of complex Chinese medicine formulas.

Chemical constituents from rhizome of Anemone amurensis.

Phytochemical investigation of the 70% EtOH extract of the rhizome of Anemone amurensis led to the isolation of two new oleanane-type triterpenoid saponins 1 and 2. Their structures were elucidated on the basis of chemical and spectral analysis, including 1D, 2D NMR data, and HR-ESI-MS. Compounds 1 and 2 were tested for cytotoxicities against two human cancer cell lines (A549 and Hep-G2). Compound 2 showed potent cytotoxicity with IC50 values of 38.53 and 66.17 µM, respectively, while compound 1 with IC50 > 100 µM.

Separation and detection of minor constituents in herbal medicines using a combination of heart-cutting and comprehensive two-dimensional liquid chromatography.

Herbal medicines contain a large number of minor constituents, which could contribute to their therapeutic effects and provide valuable lead compounds for drug discovery. However, to explore minor constituents from complicated herbal extracts is usually laborious and time-consuming. In order to discover minor novel herbal constituents efficiently, we combined heart-cutting and comprehensive two-dimensional liquid chromatography (HC-2DLC) to remove major components from herbal extracts, and then characterized the minor ones by mass spectrometry. This strategy was employed to analyze Pueraria lobata and Pueraria thomsonii, the roots of which are used as the Chinese herbal medicine Ge-Gen. Five major compounds in Ge-Gen extract were removed by on-line heart-cutting, and the minor compounds were separated on an RP×RP 2DLC system (1D, Acquity CSH C18, 2.1×100mm, 1.7µm; 2D, Poroshell Phenyl-Hexyl, 3.0×50mm, 2.7µm). A synchronized gradient elution program was used to improve chromatographic resolution of the second dimension. By using this 2DLC system, a total of 271 and 254 peaks were separated in P. lobata and P. thomsonii within 35min, respectively. The practical and effective peak capacity was 1593 and 677, respectively, and the orthogonality was around 70%. Structures of 12 selected compounds were tentatively characterized by mass spectrometry, and 9 of them were discovered from Ge-Gen for the first time. Contents of these minor compounds in Ge-Gen were preliminarily determined to be 0.01-0.1% (w/w). The HC-2DLC/MS system is a powerful and convenient tool to explore minor novel chemical constituents from complex herbal extracts.

In vivo metabolites and plasma exposure of TongMai Keli analyzed by UHPLC/DAD/qTOF-MS and LC/MS/MS.

ETHNOPHARMACOLOGICAL RELEVANCE: TongMai Keli (TM) is a widely used traditional Chinese medicine preparation for the treatment of cardiovascular and cerebrovascular diseases. It is composed of Puerariae Lobatae Radix (roots of Pueraria lobata (Willd.) Ohwi), Salviae Miltiorrhizae Radix (roots of Salvia miltiorrhiza Bge.), and Chuanxiong Rhizoma (rhizomes of Ligusticum chuanxiong Hort.). The aim of this study is to identify the in vivo metabolites of TM, and to elucidate the pharmacokinetics of TM constituents and their metabolites. MATERIALS AND METHODS: For metabolites identification, TM was orally administered to rats (n=3), and the metabolites in plasma were identified by UHPLC/DAD/qTOF-MS analysis and ß-glucuronidase hydrolysis. For pharmacokinetic study, rats (n=10) were treated with TM at a clinical dose, and the plasma was analyzed by LC/MS/MS. RESULTS: A total of 25 metabolites from TM were identified in rats plasma. Glucuronide and sulfate conjugations were the major metabolic reactions, and produced 14 metabolites. The analytical method for pharmacokinetic study was fully validated with good linearity (r>0.99), wide dynamic ranges (6-6000 ng/mL), and low variations (<14.3%). The plasma concentration-time curves of puerarin and nine metabolites were profiled. CONCLUSION: Isoflavones from Puerariae Lobatae Radix were the major metabolites in rat plasma after oral administration of TM. Puerarin and other isoflavone glycosides could reach their first C(max) within 30 min, and were then rapidly eliminated, followed by their phase II metabolites.