Molecular cloning and characterization of a novel isoflavone reductase-like gene (FcIRL) from high flavonoids-producing callus of Fagopyrum cymosum.

Lignans are important defensive compounds in plants and have good biological activities protecting human health. In order to study the medicinal secondary metabolism of Fagopyrum cymosum (Trev.) Meisn, a traditional Chinese medicine with anti-tumor effect, a novel isoflavone reductase-like gene, FcIRL, was cloned using RACE strategy from a cDNA library of high flavonoids-producing callus. The full-length cDNA of the FcIRL was 1 217 bp (accession no. EU116032), which contained a 942 bp open reading frame (ORF) encoding a 313 amino acid protein. Two stop codons (TAG) and a putative polyadenylation signal ATAAA at 24 bp upstream from the polyadenylation site was found in 5' and 3' UTR, separately. And no intron was found in the genomic sequence yet. FcIRL contained a predicted N-terminal acetylation site (M1-K5) and a NADPH-binding motif (G10-G-T-G13-Y-I-G16) in the N-terminal region, a conserved NmrA (nitrogen metabolite repression regulator) domain (V6-N244), multi-phosphorylation sites and one conserved N-glycosylation site (N214). Sequence homology comparison, phylogenetic analysis and advanced structures prediction all suggested that FcIRL belonged to the class of pinoresinol-lariciresinol reductase (PLR), which is a key enzyme in synthetic pathway of 8-8'-linked lignans, with function in catalyzing reduction of pinoresinol and lariciresinol into secoisolariciresinol, and medicinal secondary metabolism and resistance in F. cymosum.

[Impacts on vertebral arterial blood flow of cervical spondylosis of vertebral artery type treated by abdominal acupuncture].

OBJECTIVE: To explore the therapeutic effect and mechanism of abdominal acupuncture for cervical spondylosis of vertebral artery type. METHODS: Thirty cases of cervical vertigo, in which the color ultrasonography indicated vertebral arterial blood insufficiency, were treated with abdominal acupuncture therapy. The points were Zhongwan (CV 12), Qihai (CV 6), Guanyuan (CV 4), Xiawan (CV 10), Shangqu (KI 17) and Huaroumen (ST 24). The treatment was given once every day and five continuous treatments made one session. Separately, before treatment and in the 1st and 5th treatments, the cervical vertigo symptom and functional assessment scales were adopted for scoring. Simultaneously, the color ultrasonography was applied to observe the blood flow changes of the bilateral cervical arteries. RESULTS: Except the score for headache in the 1st treatment, the scores in cervical vertigo and function assessment scale in the 1st and 5th treatments were all improved significantly in 30 patients as compared with those before treatment (P < 0.01, P < 0.05). In the 1st and 5th treatments, on the affected side, the vertebral artery diameter, mean velocity and blood flow per minute were all improved significantly as compared with those before treatment (all P < 0.01). In one session treatment, the total effective rate was 100.0% (30/30) and the curative rate was 60.0% (18/30). CONCLUSION: Abdominal acupuncture therapy not only relieves the clinical symptoms, but also improves vertebral arterial blood supply for the patients of cervical spondylosis of vertebral artery type.