AIMTo study the effect of atropine, muscarinic cholinergic antagonist, on the central analgesic action of melatonin (MT) and to explore the mechanism of MT analgesia.

METHODSAs an indicator of visceral pain, the unit discharges of the neurons in the posterior group of thalamic nuclei (PO) were caused by stimulating the great splanchnic nerve (GSN) of the cat. The cranial stereotaxic and extracellular glass microelectrode record technique were used. The drugs were given through the intra-cranial-ventricle (icv).

RESULTS0.1% MT (10 micrograms.kg-1, icv) was shown to inhibit the unit discharge of the neurons in PO of the cat, whether the long latency or the short latency, which was evoked by stimulating GSN. The inhibition of 0.1% MT (10 micrograms.kg-1, icv) on the short latency discharge of neurons in PO was antagonized by 0.1% atropine (20 micrograms, icv). However, 0.1% atropine (20 micrograms, icv) did not show antagonistic effect on the inhibition of 0.1% morphine (5 micrograms, icv) at the same latency.

CONCLUSIONMT exhibited central analgesic action with mechanism different from morphine. It was suggested that the cholinergic system may be involved in analgesic process of MT.

Analgesics ; administration & dosage ; pharmacology ; Animals ; Atropine ; pharmacology ; Cats ; Electric Stimulation ; Evoked Potentials ; drug effects ; Female ; Injections, Intraventricular ; Male ; Melatonin ; administration & dosage ; pharmacology ; Morphine ; pharmacology ; Muscarinic Antagonists ; pharmacology ; Neurons ; physiology ; Splanchnic Nerves ; physiology ; Thalamic Nuclei ; drug effects ; physiology

Objective: Cimicifuga dahurica (Ranunculaceae) has many bioactivities. Although there have been intensive studies for saponin constituents at present, only a few studies have focused on for chemical constituents of phenolic acid. To define the phenolic acid constituents,C. dahurica was separated, and the structures of the compounds were identified,in the expectation of providing a basis for its further development,utilization and quality control. Method: A total of 16 kg rhizome of C. dahurica was extracted with 70%ethanol for three times by heating reflux. These 3 extracts were decompressed and concentrated,and then dissolved in water. Then the solvent was successively extracted with petroleum ether,ethyl acetate(EtOAc) and n-butanol(BuOH). The components of EtOAc and water extract were isolated and purified by macroporous,silica gel,ODS,Sephadex LH-20 column chromatography,preparative HPLC and recrystallization,and the structures were identified by nuclear magnetic resonance(NMR) and physicochemical analysis etc. Result: Fifteen compounds were isolated from the ethyl acetate and water fractions,and identified as cimicifugic G (1),2-caffeoyl piscidic acid (2),cimicifugic A (3),cimicifugic B (4),caffeic acid 3-O-β-D-glucopyranoside (5),cimicifugic E (6),cimicifugic F (7),trans-ferulic acid 4-O-β-D-glucopyranoside (8),carboxymethyl isoferulate (9),3,4-dimethoxycinnamic acid (10),ethyl ferulate (11),caffeic ester glucoside (12),shomaside A (13),isoferulic acid (14),caffeic acid (15). Conclusion: Compounds 1-7,9-10,13 were isolated from the plant for the first time.

Chondroitinase has been used as an important tool in the study of the structure, function and distribution of glycosaminoglycans for many years. Recently, the enzyme has been reported to be a potential enzyme for chemonucleolysis, an established treatment for intervertebral disc protrasion. In this paper, a chondroitinase had been purified from the culture supernatant of Aeromonas sobria YH311 using a simple purification procedure of ammonium sulfate precipitation, QAE-Sephadex A50 ion exchange chromatography and Sephadex G-150 gel filtration. The immobilization of purified chondroitinase using sodium alginate or cellulose as carriers has also been studied. The chondroitinase obtained from Aeromonas sobria YH311 was purified 55-fold to 95.3% pure, the specific activity of the purified enzyme was 31.86u/mg and the yield was 37%. The molecular weight of chondroitinase from Aeromonas sobria YH311 was determined by SDS-PAGE to be 80kD, which was almost the same as those chondroitinase AC from Arthrobacter aurescens, Aeromonas liquefaciens and Flavobacterium heparinum. But its isoelectric point was 4.3 - 4.6, which was far lower than the microbial chondroitinase AC. After the immobilization on sodium alginate or cellulose, the properties of chondroitinase changed greatly. The optimum temperature and pH of the free enzyme were 50 degrees C and 7.0 respectively, and about 10% activity remained after heat treatment at 80 degrees C for 20 minutes, and 47% activity remained after two weeks storage at 4 degrees C. The chondroitinase immobilized on sodium alginate had the optimum temperature and pH of 40 degrees C and 7.0 respectively, about 50% activity remained after 80 degrees C heat treatment for 120 minutes and 50% remained after 30 days storage at 4 degrees C. The chondroitinase immobilized on cellulose had the optimum temperature and pH of 70 degrees C and 6.0 respectively, and more than 70% activity remained after heat treatment at 80 degrees C and 30 days storage at 4 degrees C. The yield of the immobilization was very low, with 18.56% for alginate and 18.86% for cellulose.
Aeromonas ; enzymology ; Chondroitinases and Chondroitin Lyases ; isolation & purification ; metabolism ; Enzyme Stability ; Enzymes, Immobilized ; metabolism ; Temperature

Ocotillol (OT)-type ginsenosides, one subtype of ginsenosides, consist of a dammarane skeleton and a tetrahydrofuran ring. Most naturally-occurring OT-type ginsenosides exist in Panax species, particularly in Panax quinquefolius, which may be attributed to the warm and humid climate of its native areas. Till now, merely 28 types of naturally-occurring OT-type ginsenosides have been isolated. In contrast, semi-synthesized OT-type ginsenosides are attracted considerable attentions. These ginsenosides can be obtained through oxidation and cyclization of side chains of dammarane-type ginsenosides, and other methods, which may change their physical and chemical properties and further improve their bioavailabilities. It is also notable that the pharmacological activities of ginsenosides are closely related to the stereoisomers caused by the configuration at C-20. Semi-synthesis of OT-type ginsenosides can facilitate our understanding of the biosynthesis, transformation and metabolism of OT-type ginsenosides in the body. This review will systematically summarize the research progress on naturally-occurring and semi-synthetic OT-type ginsenosides, which provides a theoretical basis for their bioactivity-guided research.

OBJECTIVETo study the effects of valproate sodium (VPA) on the level and axle of pituitary gonadotropin in adolescent girls with epilepsy.

METHODSTwenty-three adolescent girls with epilepsy aged from 8 to 14 years were treated with VPA for 1 year. The levels of serum pituitary gonadotropin including estradiol, follicle-stimulating hormone, luteinizing hormone, prolactin and testosterone were measured before treatment and 3 months, 6 months and 1 year after treatment.

RESULTSThe serum levels of estradiol, follicle-stimulating hormone, luteinizing hormone and prolactin in the children with epilepsy were not significantly different during the 1 year VPA treatment compared with pretreatment. However, the serum level of testosterone was reduced 1 year after treatment (0.4±0.3 ng/mL) compared with pretreatment (0.7±0.4 ng/mL) and 3 months after treatment (0.7±0.4 ng/mL) (P<0.05).

CONCLUSIONSVPA treatment for 1 year does not increase serum levels of androgen in adolescent girls with epilepsy, suggesting that VPA is an ideal choice of treatment for the girls.

Adolescent ; Anticonvulsants ; therapeutic use ; Child ; Epilepsy ; blood ; drug therapy ; Female ; Gonadotropins, Pituitary ; blood ; Humans ; Valproic Acid ; therapeutic use

BACKGROUND: To analyze early changes in white blood cells (WBCs), C-reactive protein (CRP) and procalcitonin (PCT) in children with multiple trauma, before secondary inflammation develops. METHODS: This single-center retrospective study collected data from patients with blunt traumatic injury admitted to the pediatric intensive care unit (PICU). According to the prognostic outcome of 28 d after admission to the PICU, patients were divided into survival group (n=141) and non-survival group (n=36). Characteristics between the two groups were compared. Receiver operation characteristic (ROC) curve analysis was conducted to evaluate the capacity of different biomarkers as predictors of mortality. RESULTS: The percentages of children with elevated WBC, CRP, and PCT levels were 81.36%, 31.07%, and 95.48%, respectively. Patients in the non-survival group presented a statistically significantly higher injury severity score (ISS) than those in the survival group: 37.17±16.11 vs. 22.23±11.24 (t=6.47, P<0.01). WBCs were also higher in non-survival group than in the survival group ([18.70±8.42]×109/L vs. [15.89±6.98] ×109/L, t=2.065, P=0.040). There was no significant difference between the survival and non-survival groups in PCT or CRP. The areas under the ROC curves of PCT, WBC and ISS for predicting 28-day mortality were 0.548 (P=0.376), 0.607 (P=0.047) and 0.799 (P<0.01), respectively. CONCLUSIONS: Secondary to multiple trauma, PCT levels increased in more patients, even if their WBC and CRP levels remained unchanged. However, early rising WBC and ISS were superior to PCT at predicting the mortality of multiple trauma patients in the PICU.

Four new prenylflavonoid glycosides, namely koreanoside H-K (1-4), together with eleven known ones (5-15) were isolated from the leaves of Epimedium koreanum Nakai. Their structures were elucidated by 1D NMR, 2D NMR, HR-ESI-MS, IR and UV. The identification of the sugar moieties was carried out by means of acid hydrolysis and HPLC analysis of their derivatives. It is worth noting that compound 3 and compound 4 were elucidated to contain fucose and quinovose moieties, which were two extremely rare sugar units from the genus Epimedium. The anti-pulmonary fibrosis activity of the new compounds was evaluated using A549 cell line. Compounds 1, 2 and 4 showed significant anti-pulmonary fibrosis activities.
Chromatography, High Pressure Liquid ; Epimedium/chemistry* ; Glycosides/pharmacology* ; Plant Extracts/pharmacology* ; Plant Leaves/chemistry*

A rapid ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometric (UPLC-ESI-MS/MS) method is developed for the qualitative identification of constituents in the flower buds of seven Lonicera species. The optimal condition of separation and detection were achieved on an AcQuity UPLC BEH C18 column with a gradient elution with acetonitrile and 0.1% acetic acid within 17 min. Among the 33 constituents detected, 6 caffeoylquinic acids (including caffeic acid), 8 flavonoids and 8 iridoid glycosides were characterized based on their fragmentation patterns in collision-induced dissociation (CID) experiments and/or by comparison with standard compounds. In addition, to statistically establish the correlation and discrimination of the Lonicera species, principle component analysis (PCA) was applied in this study. Lonicera samples were divided into well-defined groups directly related to their species based on PCA in terms of the log transformed relative contents of the major caffeoylquinic acids (including caffeic acid) as the variables. All of results indicated that the method presented here is able to classify the sample species and to reveal characteristic details of the chemical constituents of different Lonicera species.
Chromatography, Liquid ; methods ; Flowers ; chemistry ; Lonicera ; chemistry ; classification ; Principal Component Analysis ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods

AIMTo observe the role of melatonin receptor and GABAA receptor in sleeping time prolonged by melatonin in mice.

METHODSThe absence of the righting reflex was considered as the sleep onset and the duration of the loss of the righting reflex was recorded as the sleeping time. The effects of receptor agonist and antagonist on hypnotic activity of melatonin were studied in the paper.

RESULTSPrazosin hydrochloride, the blocker of melatonin 3 receptor, didn't affect the sleeping time prolonged by melatonin in mice. GABA, the endogenous agonist of GABA receptor, significantly potentiated the hypnotic activity of melatonin. When picrotoxin, the ligand of picrotoxin site on GABAA receptor, used together with melatonin, it significantly antagonized the sleeping time prolonged by melatonin, however, bicuculline, the specific antagonist of GABA binding site in GABAA receptor, didn't affect the hypnotic activity of melatonin in mice.

CONCLUSIONMelatonin does not exhibit its potentiation sleeping time in mice through melatonin 3 receptor. Hypnotic activity of melatonin may be mediated through picrotoxin site on GABAA receptor.

Animals ; Bicuculline ; pharmacology ; Male ; Melatonin ; physiology ; Mice ; Mice, Inbred Strains ; Picrotoxin ; pharmacology ; Prazosin ; pharmacology ; Receptors, GABA-A ; physiology ; Receptors, Melatonin ; physiology ; Sleep ; physiology

Structure of natural product-ursolic acid was modified for increasing its antitumor activity. Ursolic acid was acylated, esterified, hydrolized or oxidized to obtain target pentacyclic triterpenoid compounds with different substitutes. Sixteen derivatives of ursolic acid were designed and synthesized including eleven new compounds. Anti-tumor activities of ursolic acid and these derivatives against HeLa, SKOV3 and BGC-823 cells in vitro were investigated by MTT assay. The results indicated that compounds 7a and 8a were found to have stronger cell growth inhibitory than ursolic acid on HeLa cells and SKOV3 cells separately, and are worth to be intensively studied further.
Antineoplastic Agents, Phytogenic ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; HeLa Cells ; Humans ; Triterpenes ; chemical synthesis ; chemistry ; pharmacology