OBJECTIVETo evaluate the inhibitory effect of total bufadienolides from toad venom against H22 tumor in mice and preliminarily analyze the structures of the metabolites in tissues.

METHODHPLC and LC-MS were used for analysis of the chemical composition of TBFs. High, middle and low dosages of TBFs were orally administered or intra-peritoneally injected to H22 tumor-bearing mice for thirteen days. The animals were killed and the tumors were stripped and weighed. The metabolites in the tissues such as heart, liver, spleen, lung and kidney, were analyzed by HPLC and LC-MS.

RESULTThe chemical composition of TBFs were identified by comparison of the retention times with those of reference substances, on-line UV spectra and MS data. Its main components are concerned with gamabufotalin, arenobufagin, bufotalin, resibufagin, cinobufotalin, bufalin, cinobufagin and resibufogenin. TBFs had no obvious influence on body weight of H-22 tumor-bearing mice orally administered and the inhibition rate against tumor were 14.76%, 16.38% and 10.32% for low (5 mg x kg(-1)), middle (10 mg x kg(-1)) and high dosage (20 mg x kg(-1)), respectively. The mice intra-peritoneally injected with middle and high-dose of TBFs gained body weight slower than the control mice on the 5th day and recovered on the 13th day. The inhibition rate against tumor were 17.30%, 19.80% and 40.95% for low (1.5 mg x kg(-1)), middle (3 mg x kg(-1)) and high dose (6 mg x kg(-1)), respectively. The inhibitory effect took on dose-dependent manner. Based on the HPLC analyses on heart, liver, spleen, lung and kidney, bufadienolides were found in the liver tissue and 11 compounds of them were tentatively identified by LC-DAD-MS.

CONCLUSIONTBFs by oral administration had no inhibitory effect against H22 tumor in mice, however, TBFs by intra-peritoneal injection displayed the significantly inhibitory effect, accompanying some toxicity for early duration of the study. The identification of bufadienolides in the liver provides a good basis for the further investigation of the metabolic pathways of TBFs in vivo.

Amphibian Venoms ; chemistry ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; metabolism ; Bufanolides ; administration & dosage ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; physiopathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Administration Routes ; Female ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Neoplasm Transplantation

Eight compounds were isolated from Bufonis periostracum by repeated column chromatography on silica gel, ODS and Sephadex LH-20 and their structures were characterized as palmitatic acid cholesteryl ester (1), cholesterol (2), 5alpha, 8alpha-epidioxycholesta-6-en-3beta-ol (3), cholest-5-en-3beta, 7beta-diol (4), cholest-7-en-3beta, 5alpha, 6beta-triol (5), 3-octaddecyloxy-1, 2-propanediol (6), isisamide (7) and bufothionine (8) on the base of spectral analysis. Compounds 1-8 were isolated from Bufonis periostracum for the first time and compounds 3, 5, 6, 7 were obtained from Bufo bufo gargarizans and Bufo genus for the first time. The bioassays showed all tested samples displayed no antitumor activity against the cell lines such as A549, BeL 7402, HGC-27 and HL-60, except the control compound bufalin.
Animals ; Antineoplastic Agents ; pharmacology ; Bufo bufo ; metabolism ; Bufonidae ; metabolism ; Humans

Children with specific language impairment(SLI)have normal cognitive abilities but experience language deficits.The heterogeneity of language disorders makes SLI children optimal subjects for investigating the correlation between language ability and theory of mind.This paper provides a comprehensive review of foreign em-pirical studies examining the relationship between language ability and theory of mind in SLI groups from various perspectives.The findings indicate that specific types of language disorders exhibit delayed development in theory of mind,which is closely associated with their language ability deficits.Among all linguistic elements,syntactic de-fects demonstrate the most concentrated evidence regarding their impact on theory of mind.It is noteworthy that al-though SLI children offer valuable insights into the correlation between language and theory of mind,further investi-gation is still required to comprehensively comprehend this association.The present study also provides recommenda-tions for future research endeavors in this domain.