Optimization of Ultrasound-Assisted Extraction, HPLC and UHPLC-ESI-Q-TOF-MS/MS Analysis of Main Macamides and Macaenes from Maca (Cultivars of Lepidium meyenii Walp).

Ultrasound-assisted extraction (UAE), using petroleum ether as the solvent, was systematically applied to extract main macamides and macaenes from Maca hypocotyls. Extraction yield was related with four variables, including ratio of solution to solid, extraction temperature, extraction time, and extraction power. On the basis of response surface methodology (RSM), the optimal conditions were determined to be the ratio of solution to solid as 10:1 (mL/g), the extraction temperature of 40 °C, the extraction time of 30 min, and the extraction power of 200 W. Based on the optimal extraction method of UAE, the total contents of ten main macamides and two main macaenes of Maca cultivated in twenty different areas of Tibet were analyzed by HPLC and UHPLC-ESI-Q-TOF-MS/MS. This study indicated that UAE was able to effectively extract macamides alkaloids from Maca hypocotyls. Quantitative analysis showed that geographical origins, not ecotypes, played a more important role on the accumulation of active macamides in Maca.

[Effects of acupuncture-moxibustion intervention on proprioception in athletes with lateral collateral ligament injury of ankle joint].

OBJECTIVE: To compare the efficacy differences between acupuncture-moxibustion and physiotherapy interventions in improving proprioception of athletes with lateral collateral ligament injury of the ankle joint. METHODS: Thirty patients with injured lateral collateral ligament of ankle joint were randomly divided into acupuncture group (n = 15) and physiotherapy group (n = 15). Patients of the acupuncture group were treated by acupuncture and moxibustion stimulation of Qiuxu (GB40), Kunlun (BL 60), Shenmai (BL 62), Jiexi (ST 41), and Ashi-points, etc., and those of the physiotherapy group treated with TDP irradiation of the regional lateral malleolus. The treatment of the two groups was conducted once the other day, 3 times each week, continuously for 8 weeks. Before and after the treatment, the ankle-joint's active and passive repositioning error angles were measured by using a joint angle ruler. RESULTS: The average error angle values of active and passive reposition tests of the injured ankle-joint were 4.98 +/- 1.11 and 4.78 +/- 1.3 before the treatment, and 3.67 +/- 0.58 and 3.51 +/- 0.64 after the treatment, respectively in the acupuncture group, being reduced significantly after the treatment (both P < 0.01). No significant changes of the average error angle values of both active and passive reposition tests of the ankle-joint were found after the treatment in the physiotherapy group (P > 0.05). Comparison between two groups showed that the average error angle, average active and passive reposition angles of the injured ankle in the acupuncture group were evidently lower than those in the physiotherapy group (P < 0.01). CONCLUSION: Acupuncture and moxibustion can effectively improve the proprioception of the injured lateral collateral ligament of the ankle joint in athletes, which is superior to conventional physiotherapy in the therapeutic effect.

[Effects of sinomenine on synovial fluid and serum content of interleukin-1beta in rabbits with osteoarthritis].

OBJECTIVE: To investigate the effects of intra-articular injection of sinomenine on interleukin-1beta (IL-1beta) content in synovial fluid and serum in rabbits with osteoarthritis (OA). METHODS: Degenerative osteoarthritis in the knee joint of left posterior limb was induced in 36 rabbits by full extension using plaster cast for 6 weeks. Then the rabbits were randomly divided into untreated group, hyaluronic acid group and sinomenine group. Another 4 normal rabbits were selected as normal control group. Rabbits in the sinomenine group and the hyaluronic acid group received intraarticular injections of sinomenine and hyaluronic acid once a week for 5 weeks, respectively. The content of IL-1beta in synovial fluid and serum were measured before and after treatment by enzyme-linked immunosorbent assay. The pathological changes of cartilaginous tissue were analyzed by using Mankin's score. RESULTS: Compared with those in the untreated group, synovial fluid and serum IL-1beta contents in the sinomenine group and the hyaluronic acid group were significantly decreased (P<0.01). And the synovial fluid and serum IL-1beta contents in the sinomenine group were lower than those in the hyaluronic acid group (P<0.05). The mean Mankin's score of cartilage in the sinomenine group was significantly lower than that in hyaluronic acid group (P<0.05). CONCLUSION: Sinomenine may reduce the degree of articular degeneration in rabbit with OA through decreasing the content of IL-1beta in synovial fluid and serum.

In vitro anti-HCV activities of Saxifraga melanocentra and its related polyphenolic compounds.

The aim of this study was to search for new natural anti-HCV agents from Chinese herbal medicine. Bioactivity-guided extraction and isolation methods were used. Active part and pure compounds were obtained from ethanolic extract of Saxifraga melanocentra Franch. and their in vitro inhibitory activities (IC50) against HCV NS3 serine protease were tested by enzyme-linked immunosorbent assay. Results showed that the polyphenolic ethyl acetate part of the herbal extract was the most active, and from this 18 polyphenols representing active compounds were isolated and identified. IC50 values of these compounds and five related ones were obtained. A broad-degree of anti-HCV activity was observed among them in the following order: gallated esters of D-glucose and rutin (0.68-4.86 microM)> flavonoids (33.11-370.37 microM)> gallic acid and its methyl and ethyl esters, Bergenin and others (over 1000 microM). The most active compound was 1,2,3,4,6-penta-O-galloyl-beta-D-glucoside (0.68 microM). In conclusion, polyphenols were responsible for the anti-HCV constitution of S. melanocentra, and multigallated esters of D-glucose possessed the strongest inhibition against HCV NS3 serine protease and little cytotoxic effect, suggesting the potential use of these compounds for designing and developing drugs for treatment of the viral infection.

[Histopathological changes in rat transplanted hepatoma after lipiodol transarterial embolization].

OBJECTIVE: To study the histopathological effect of hepatic arterial infusion of lipiodol on transplanted hepatoma in rats. METHODS: Fourty-one rats bearing Walker-256 transplanted hepatoma were randomly divided into embolization group (n = 35, divided in 5 subgroups, with 7 rats in each) and control group (n = 6). Lipiodol (0.5 ml/kg)emulsified with 0.2 - 0.3 ml of 76% urografin (v:v = 1:1) was infused via gastroduodenal artery into hepatic artery in embolization group. Rats in the control group were given via the same route urografin only. Histopathological changes of the treated tumors were examined by light and transmission electron microscopy. RESULTS: In the control rats treated with urografin alone, the average tumor size increased 2.8 fold on day 3, while that in the lipiodol treated rats increased 1.7 fold (P < 0.01). Compared with the control group, on day 3, 5, 10 after embolization treatment, tumor necrosis was more extensive (P < 0.01). In one of the treated rats, the tumor was completely necrotic on day 10. Inflammatory reaction was marked in the early post-embolic period, but it was replaced by fibrous tissue encapsulation. From day 1 on, in 17 of the 18 treated rats, apoptotic cells, identified by typical morphology under light and electronic microscopes, were observed, mainly in the tumor periphery. CONCLUSION: In addition to cellular necrosis, apoptosis may be another important mechanism leading to cell death in hepatoma treated with transarterial embolization.