Expression profiling and functional verification of flavonoid 3'-hydroxylase gene from leaves of Euryale ferox / 中国中药杂志

Leaves of Euryale ferox are rich in anthocyanins. Anthocyanin synthesis is one of the important branches of the flavonoid synthesis pathway, in which flavonoid 3'-hydroxylase(F3'H) can participate in the formation of important intermediate products of anthocyanin synthesis. According to the data of E. ferox transcriptome, F3'H cDNA sequence was cloned in the leaves of E. ferox and named as EfF3'H. The correlation between EfF3'H gene expression and synthesis of flavonoids was analyzed by a series of bioinforma-tics tools and qRT-PCR. Moreover, the biological function of EfF3'H was verified by the heterologous expression in yeast. Our results showed that EfF3'H comprised a 1 566 bp open reading frame which encoded a hydrophilic transmembrane protein composed of 521 amino acid residues. It was predicted to be located in the plasma membrane. Combined with predictive analysis of conserved domains, this protein belongs to the cytochrome P450(CYP450) superfamily. The qRT-PCR results revealed that the expression level of EfF3'H was significantly different among different cultivars and was highly correlated with the content of related flavonoids in the leaves. Eukaryotic expression studies showed that EfF3'H protein had the biological activity of converting kaempferol to quercetin. In this study, EfF3'H cDNA was cloned from the leaves of E. ferox for the first time, and the biological function of the protein was verified. It provi-ded a scientific basis for further utilizing the leaves of E. ferox and laid a foundation for the further analysis of the biosynthesis pathway of flavonoids in medicinal plants.

Optimization of combined enzymatic extraction for alkaloids and polysaccharides from Dendrobium nobile / 中成药

AIM To optimize the combined enzymatic extraction for alkaloids and polysaccharides from Dendrobium nobile Lindl..METHODS With enzyme consumption,enzymolysis temperature,enzymolysis time and solidliquid ratio as influencing factors,contents of dendrobine,total alkaloids and polysaccharides as evaluation indices,orthogonal test was applied to optimizing the combined enzymatic extraction.RESULTS The optimal conditions for papain extraction were determined to be 0.10 g for enzyme consumption,45 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶ 50 for solid-liquid ratio,the contents of dendrobine,total alkaloids and polysaccharides were 3.495 5,4.341 8 and 35.898 7 mg/g,respectively.The optimal conditions for cellulase extraction were determined to be 0.30 g for enzyme consumption,50 ℃ for enzymolysis temperature,2 h for enzymolysis time,and 1 ∶40 for solid-liquid ratio,the contents of three constituents were 3.514 8,4.351 3 and 36.331 2 mg/g,respectively.The optimal conditions for pectinase extraction were determined to be 0.45 g for enzyme consumption,55 ℃ for enzymolysis temperature,2.5 h for enzymolysis time,and 1 ∶ 40 for solid-liquid ratio,the contents of three constituents were 3.524 4,4.452 8 and 26.324 2 mg/g,respectively.CONCLUSION This stable and reliable method can be used for the rapid combined enzymatic extraction for alkaloids and polysaccharides from D.nobile.

Discussion on the regular of acupoint selection of acupuncture and moxibustion for lumbar disc herniation during recent 10 years / 中国针灸

By taking "lumbar disc herniation", Chinese medicine", "acupuncture and moxibustion" as key words Chinese articles about acupuncture for lumbar disc herniation in recent 10 years were searched in three major Chinese databases of Wan fang database, VIP database and CNKI. Totally 546 relative articles were retrieved. After the elimination of invalid articles, 173 were included as valid articles with 49 main acupoints. The hierarchical clustering statistical method was used to analyze the possible rules among the former 20 acupoints. It turned out that there were 10 most common used acupoints on Bladder Meridian [Geshu (BL 17), Xiaochangshu (BL 27), Shenshu (BL 23), Dachangshu (BL 25), Pangguangshu (BL 28), Guanyuanshu (BL 26), Weizbong (BL 40), Zhibian (BL 54), Chengshan (BL 57), Kunlun (BL 60)], 2 on Gallbladder Meridian [Huantiao (GB 30), Yanglingquan (GB 34)], 2 on Governor Vessel [Yaoyangguan (GV 3), Shuigou (GV 26)], 2 on Stomach Meridian [Zusanli (ST 36), Juliao (ST 3)], 2 on Spleen Meridian [Sanyinjiao (SP 6), Xuehai (SP 10)], and the rest were extra points (Huatuo Jiaji) and Ashi points, so a conclusion could be drawn that the most common used acupoints were Bladder Meridian acupoints and supplemented by Gallbladder Meridian, Governor Vessel, Stomach Meridian, Spleen Meridian, extra points and Ashi points. The selected acupoints were most located on the lumbosacral region, leg and fewer located on the face, back and local part.

Risk factors and pain status due to diabetic neuropathy in chronic long-term diabetic patients in a Chinese urban population / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>With economic growth and urbanization there have been significant changes in the life style and diet of urban residents in large cities of China, which is experiencing a rapid increase in the prevalence of diabetes. While high prevalence of diabetes has been reported, little is known of the long-term effects of diabetes in such a large population. The aim of this study was to estimate the morbidity rate of diabetic peripheral neuropathy (DPN) in a Chinese urban diabetic population with more than 10 years' disease duration, and evaluate the relevant risk factors. The clinical manifestation of DPN and pain status was also assessed.</p><p><b>METHODS</b>Five hundred and sixty-five diabetes patients were recruited into the study. Symptoms and examination helped diagnose neuropathy. The clinical manifestation of DPN was assessed with a visual analog pain score (VAS). Diabetic complication status was determined from medical records. Serum lipids and lipoproteins, glycosylated hemoglobin (HbA1c), and the urinary albumin excretion rate were measured.</p><p><b>RESULTS</b>The morbidity rate of DPN was 46.6%. HbA1c, hyperlipidemia, and retinopathy were significantly associated with neuropathy, and these risk factors were correlated with other diabetic micro and/or macrovascular complications. The average VAS pain score of the DPN patients was 4.12 ± 2.07. Severe and moderate pain was experienced by 11.4% and 40.5% respectively of DPN patients. About 3.7% of diabetic subjects had lower limb ulcer or amputation.</p><p><b>CONCLUSIONS</b>The morbidity rate of DPN for diabetic patients with > 10 years duration is very high compared to the range reported for other populations in the world. The risk factors for DPN include HbA1c, hyperlipidemia, and retinopathy. In long-standing diabetic patients, DPN was not associated with diabetic duration, and half of the DPN patients experienced considerable daily suffering.</p>