Objective To investigate the anti-HBV constituents from Alternanthera philoxeroides.Methods The constituents were isolated with silica gel and gel permeation chromatography,and purified by HPLC.Their structures were elucidated by spectroscopy.The antivirus effects of the isolated compounds were tested by ELISA method in vitro.Results Ten compounds were isolated and elucidated as followings:oleanolic acid(Ⅰ),oleanolic acid 3-O-?-D-glucuronopyranoside(Ⅱ),oleanolic acid 28-O-?-D-glucopyranoside(Ⅲ),chikusetsusaponin Ⅳ a methyl ester(Ⅳ),4,5-dihydroblumenol(Ⅴ),N-trans-feruloyl 3-methyldopamine(Ⅵ),N-trans-feruloyl tyramine(Ⅶ),3?-hydroxystigmast-5-en-7-one(Ⅷ),24-methylenecycloartanol(Ⅸ),and cycloeucalenol(Ⅹ).The values of inhibition percent of compounds Ⅰ-Ⅲ,Ⅴ-Ⅶ revealed a significant distinction compared to the control group.Compounds Ⅱ and Ⅴ showed significant inhibition against HepG2 cells transected with cloned hepatitis B virus DNA,their inhibitive ratios were 85.38% and 87.37% at 50 ?g/mL,respectively.Conclusion Compounds Ⅳ-Ⅶ are isolated from this plant for the first time and phenolic amides have been determined as the new structure type from the plants of Alternanthera Forsk.Compounds Ⅱ and Ⅴ from A.philoxeroides show the more significant anti-HBV activities.

BACKGROUND:Cortical electrical stimulation has achieved good effects in treatment of stroke through animal and clinical experiments.
OBJECTIVE:To observe the effects of a ful y implanted cortical electrical stimulation device with long time, low intensity and various frequencies stimulation protocols on the neurological function recovery in a rat model of local cerebral infarction.
METHODS:The cerebral infarction model was established through middle cerebral artery occlusion in 60 Sprague-Dawley adult male rats. Forty rats with 1-3 points by Bederson scale were detected with magnetic resonance imaging, which was used to confirm cortex infarction and to identify a location for implantation of stimulating electrode over the peri-infarct cortex. Twenty-three rats with cortex infarction were randomly divided into cortical electrical stimulation group (CES group, n=13) and no stimulation group (NS group;n=10). The device was implanted on 6 days after middle cerebral artery occlusion, and the stimulation was given for 16 days. The stimulation program consists of two sessions lasting half an hour each in the morning and in the afternoon respectively. Stimulator delivered biphasic charge balanced pulses (pulse width=200μs) with various frequencies of 50 Hz, 20 Hz and 5 Hz within 10 second blocks and then repeated. The rats of NS group were implanted with the device, but received no electrical stimulation. The behavioral tests, includingforelimb use asymmetry test and foot fault test were performed at 2 and 16 days after implantation. Final y, al of the devices were taken out to test if they were normal y working and al of the rats were sacrificed for hematoxylin-eosin staining, which can reflect the structure of peri-infarct cortex and cellmorphology.
RESULTS AND CONCLUSION:There was only one stimulator in CES group cannot normal y work, and the remaining 22 ones worked wel . The skin covered the implanted stimulator was slightly ulcerated in one rat, and the incisions of the other rats were healed wel . Hematoxylin-eosin staining showed clear and intact structure in peri-infarction cortex (i.e., electrodes were implanted at the cortex), neurons arranged in neat rows, with abundant neuronal cytoplasm and clear nucleolus. The glial cells have complete structures, and there was no edema in the intercellular spaces. Foot-fault and forelimb use asymmetry tests showed the improved neurological function in rats of CES group than that of NS group. We designed a ful-implanted cortical electrical stimulator used in cerebral ischemic rats, and established an implanted method with long time, low intensity and various frequencies pulsed electrical stimulation. The results indicated the stimulation pattern in our study is safe and effective, and it can significantly promote functional recovery in local cerebral infarction rats.

Objective To study the diterpenoids possessed immunosuppressive activity from Tripterygium hypoglaucum. Methods The chemical constituents were isolated from the CHCl3 extracts of T. hypoglaucum by repeated column chromatography, including silica gel, Sephadex LH-20, and preparative HPLC. Their structures were elucidated on the basis of spectroscopic studies. The immunosuppressive activity of these compounds was tested using the lymphocyte transformation test. Results Compounds Ⅰ-Ⅵ were identified as triptobenzene H (Ⅰ), triptoquinone A (Ⅱ), triptoquinone B (Ⅲ), triptoquinone H (Ⅳ), triptonediol (Ⅴ), triptonoterpene (Ⅵ). Conclusion Compound Ⅰ-Ⅲ are isolated from this medicinal plant for the first time and all the compounds show the significant immunosuppressive activity.

Objective To investigate the effect of implanted cortical electrical stimulation (CES) on the expression of Nissl bodies and growth-associated protein 43 (GAP-43) in the brain after ischemic injury,and its mechanism.Methods Models of middle cerebral artery occlusion (MCAO) were established in 23 male Sprague-Dawley rats.They were randomly divided into a CES group (CES,n=13) and a no stimulation group (NS,n=10) and electrical stimulators were implanted in both groups.CES was applied for 14 d in the CES group but not in the NS group.The expression of Nissl bodies and GAP-43 around the infarct were quantified using version 6.0 of the ImagePro Plus system.Results In the CES group the Nissl bodies had a deep color,and their percentage of area was higher than that in the NS group.The GAP-43 positive expression area also had a relatively deep color,and the average percentage of positive expression area was also higher than that in the NS group.Conclusions CES can enhance the expression of Nissl bodies and GAP-43 after cerebral infarction.This suggests that CES can promote axon growth and the formation of new neural circuits.

Objective To develop an HPLC method for determination of triptoquinone H in Tripterygium wilfordii and its Tablet preparations. Methods An external method with Agilent Zorbax SC-C_(8)(250 mm?4.6 mm, 5 ?m) column as fixed phase and methanol-water (75∶25) as mobile phase was adopted. The detective wavelength was 258 nm and the flow rate was 1.0 mL/min. Results The linearity range of triptoquinone was 6.28 — 100.5 ?g/mL (r=0.999 7). The average recovery of T. wilfordii was 96.94%, RSD was 1.57% (n=9) and the average recovery of T. hypoglaucum Tablet was 100.02%, RSD was 1.74% (n=9). Conclusion The method is accurate and sensitive. It is adoptable for quantity analysis of triptoquinone H in T. wilfordii and its Tablet.

Objective To determine the content of tripterine in Tripterygium wilfordii and its tablets by HPLC.Methods An external standard method by HPLC with Zorbax C_(18)column as fixed phase and methanol-1% HAc(87∶13) as mobile phase was adopted.The detection wavelength was 425 nm and the flow rate was 1.0 mL/min.Results The linear range for tripterine was 40.96～204.8 ?g/mL(r=(0.999 6).) The average recovery of Chinese medicinal materials was 98.37% and RSD was 1.01%(n=9);the average recovery of preparation sample was 98.59% and RSD was 1.18%(n=9).Conclusion The method is simple and accurate,which can be adoptable for quantitative analysis of tripterine in the plants of Tripterygium L.

Objective To establish an HPLC method for determination of triptoquinone B in Radix Folium Seu Flos Tripterygii Wilfordii(RFFTW) and its tablets.Methods An external method with HiQ siL KYA-C_(18)(250 mm?4.6 mm,5 ?m) column as fixed phase and methanol-1% acetic acid solution((66∶)34) as mobile phase was adopted.The detective wave length was 254 nm and the flow rate was 1.0 mL/min.Results The linearity range was 6.62—105.9 ?g/mL(r=0.999 9),the average recovery of T.wilfordii from Hunan Province was 97.78%,RSD was 1.09%(n=9),and the average recovery of tablet of T.hypoglaucum was 99.83%,RSD was 1.76%(n=9).Conclusion The method is accurate and sensitive.It is adoptable for quantitative analysis of triptoquinone B in RFFTW and its tablets.

OBJECTIVETo investigate antitumor constituents from n-butyl alcohol extract of Alternanthera philoxeroides.

METHODThe constituents were isolated with silica gel, gel permeation chromatography, and purified by HPLC. Their structures were elucidated by spectroscopy and acid hydrolysis. The antitumor effects of extracts and isolated compounds were tested by MTH method in vitro.

RESULTSeven compounds were isolated and elucidated as followings: oleanolic acid 3-O-beta-D-glucuronopyranoside (1), oleanolic acid 28-O-beta-D-glucopyranoside (2), oleanolic acid 3-O-beta-D-glucuronopyranoside-6'-O-methyl ester (3), chikusetsusaponin IV a methyl ester (4), hederagenin 3-O-beta-D-glucuronopyranoside-6'-O-methyl ester (5), 4,5-dihydroblumenol (6), 6S,7E,9R-6,9-di-hydroxymegastigma-4,7-dien-3-one-9-O-beta-D-glucopyranoside (7). Compound 1 showed significant inhibitory effect against Hela and L929 with inhibitive ratios 91.3% and 92.9% at 30 mg x L(-1), respectively.

CONCLUSIONCompounds 4, 5 and 7 were isolated from this genus for the first time. Compound 1 showed significant inhibitory effect against Hela and L929 at 30 mg x L(-1).

Amaranthaceae ; chemistry ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; Hydrolysis