[Cloning of transcription factor PcFBA-1 in Pogostemon cabin and its interaction with FPPS promoter].

Farnesyl diphosphate synthase(FPPS) is a key enzyme at the branch point of the sesquiterpene biosynthetic pathway, but there are no reports on the transcriptional regulation of FPPS promoter in Pogostemon cabin. In the early stage of this study, we obtained the binding protein PcFBA-1 of FPPS gene promoter in P. cabin. In order to explore the possible mechanism of PcFBA-1 involved in the regulation of patchouli alcohol biosynthesis, this study performed PCR-based cloning and sequencing analysis of PcFBA-1, analyzed the expression patterns of PcFBA-1 in different tissues by fluorescence quantitative PCR and its subcellular localization using the protoplast transformation system, detected the binding of PcFBA-1 protein to the FPPS promoter in vitro with the yeast one-hybrid system, and verified its transcriptional regulatory function by dual-luciferase reporter gene assay. The findings demonstrated that the cloned PcFBA-1 had an open reading frame(ORF) of 1 131 bp, encoding a protein of 376 amino acids, containing two conserved domains named F-box-like superfamily and FBA-1 superfamily, and belonging to the F-box family. Moreover, neither signal peptide nor transmembrane domain was contained, implying that it was an unstable hydrophilic protein. In addition, as revealed by fluorescence quantitative PCR results, PcFBA-1 had the highest expression in leaves, and there was no significant difference in expression in roots or stems. PcFBA-1 protein was proved mainly located in the cytoplasm. Furthermore, yeast one-hybrid screening and dual-luciferase reporter gene assay showed that PcFBA-1 was able to bind to FPPS promoter both in vitro and in vivo to enhance the activity of FPPS promoter. In summary, this study identifies a new transcription factor PcFBA-1 in P. cabin, which directly binds to the FPPS gene promoter to enhance the promoter activity. This had laid a foundation for the biosynthesis of patchouli alcohol and other active ingre-dients and provided a basis for metabolic engineering and genetic improvement of P. cabin.

[Establishment of near-infrared on-line monitoring model and determination of endpoint in frying process of Hordei Fructus Germinatus].

Hordei Fructus Germinatus has been always used by "stir-frying" as a traditional medicine and the endpoint judgment of "fragrant" and "yellow" has been the focus and difficulty in frying process research. In this study, a quantitative calibration model between index components and NIRS was established in order to rapidly detect the contents of reducing sugar, total amino acids, total flavonoids, A420 and moisture; besides, an endpoint judgment method of frying process was put forward based on the "component change rate". Near-infrared spectra of samples with different frying time were collected, and a quantitative analysis model based on near-infrared spectroscopy was established to optimize the parameters such as spectral pretreatment and modeling band. HCA and PLS-DA were used to study the relationship between component and "stir-frying", and the endpoint of frying process was determined. The established calibration model had a good performance that the correlation coefficients between the predicted results and the actual measured values were both more than 0.9, with predicted relative deviations less than 10%. Hordei Fructus Germinatus with different frying time was divided into 4 categories by HCA analysis. PLS-DA analysis showed that total reducing sugar had a significant contribution to distinguishing the Hordei Fructus Germinatus of different frying time. When the change rate of total reducing sugar was 80%, it could judge that the endpoint of frying had been obtained. Results showed that NIRS could not only quickly and accurately determine the contents of reducing sugar, total amino acid, total flavonoid, A420 and moisture in Hordei Fructus Germinatus samples of different frying time, but also judge the endpoint of frying in the process. This study provided a new method for the evaluation and endpoint judgment of "stir-frying" process in research of traditional Chinese medicine.

[Observation on therapeutic effect of shu-needling therapy as main on cervical spondylosis of nerve root type].

OBJECTIVE: To explore a better method for treatment of cervical spondylosis of nerve root type. METHODS: One hundred and two cases were randomly divided into an observation group and a control group, 51 cases in each group. The observation group were treated with Shu-needling, i. e. the needles were deeply inserted into the cervical vertebrae of corresponding Jiaji (EX-B 2), and the control group were treated with routine needling method. Electroacupuncture was used in the two groups. RESULTS: The cured rate and the total effective rate were 68.6% and 98.0% in the observation group and 47.1% and 84.3% in the control group, with significant differences between the two groups (both P<0.05). CONCLUSION: Shu-needling therapy as main has a better therapeutic effect on cervical spondylosis of nerve root type.

[Observation on therapeutic effect of long-time needle retention at Baihui (GV 20) on vertebroarterial cervical spondylopathy].

OBJECTIVE: To compare therapeutic effects of long-time needle retention at Baihui (GV 20) and routine needle retention on vertebroarterial cervical spondylopathy. METHODS: One hundred and twenty cases of vertebroarterial cervical spondylopathy were randomly divided into a treatment group and a control group, 60 cases in each group. The treatment group were treated with needle retention at Baihui (GV 20) for 8 h and electroacupuncture at local points, and the control group with needle retention at Baihui (GV 20) for 30 min and electroacupuncture at local points. RESULTS: The cured rate and the total effective rate were 70.0% and 98.3% in the treatment group and 45.0% and 86.7 in the control group with very significant difference or significant difference in the cured rate and the total effective rate between the two groups (P < 0.01, P < 0.05). CONCLUSION: Long-time needle retention at Baihui (GV 20) has a better therapeutic effect on vertebroarterial cervical spondylopathy.