Chemical components of the volatile oil from leaves of Cananga odorata and its anti-oxidant activity

Cananga odorata [Lamk.] Hook. f. et Thoms., belonging to Annonaceae, is an evergreen tree. The oils extracted from its flower are a famous perfume and used in daily chemical and food industry. Although this plant has been widely cultivated in tropical regions of the world, the yield of oils from its flower is very limited. In order to develop the other parts of this plant, the chemical constituents of the volatile oils from the leaves of C. odorata was analyzed by gas chromatography/flame ionization detector [GC-FID] and GC/mass spectrometry [GC-MS]. And the volatiles showed nitric oxide [NO] inhibitory activity with an IC50 value of 37.61?g/mL and anti-oxidant activity with an IC50 value of 3.84mg/mL, respectively

Microcalorimetric investigation on the interaction of six alkaloids from rhizoma coptidis / 药学学报

How to identify active constituents of traditional Chinese medicines (TCMs) and study their interactions are key problems in the development of TCMs. The inhibitory effect of six alkaloids from Rhizoma Coptidis (RC) on Shigella dysenteriae (S. dysenteria) growth had been investigated by microcalorimetry in this study. Main active constituents of RC were confirmed by comparing their contributions to the bacteriostatic effect, and the interactions among active constituents were further researched. According to the result, in 0.8 mg-mL-1 extract of RC, the contributions of six active alkaloids including berberine, coptisine, epiberberine, palmatine and the combination of jatrorrhizine and columbamine were 52.83%, 36.31%, 2.49%, 4.27% and 3.21%, respectively. Therefore, berberine and coptisine were the main active constituents of RC that inhibited the growth of S. dysenteria. The study of interactions among the six alkaloids indicated that, 1 there were some contstituents antagonizing the inhibitory effect of RC, 2 there was a synergy effect between berberine and coptisine, 3 there were additive effects between other four alkaloids and the main active constituents. These results may provide some useful references for the establishment of the quality standard for RC and the development of multi-component TCMs.

Molecular cloning of farnesyl diphosphate synthase from Eleutherococcus senticosus and its bioinformatics and expression analysis / 中国中药杂志

<p><b>OBJECTIVE</b>To clone farnesyl diphosphate synthase (FPS) gene from Eleutherococcus senticosus and analyze the bioinformatics and expression of the gene.</p><p><b>METHOD</b>The FPS full length cDNA was cloned by rapid amplification of cDNA ends (RACE). The data was analyzed by bioinformatics method, the structure and function of FPS was deduced. The expression of FPS in different organ of E. senticosus was detected by RT-PCR.</p><p><b>RESULT</b>The full length of FPS cDNA was 1 499 bp containing a 1 029 bp ORF that encoded 342 amino acids. The deduced protein sequence exhibited two Asp riches conserved motifs (DDXXD). Without transmembrane domain, FPS was located in cytoplasm. RT-PCR result showed that FPS gene expressed in different organs of E. senticosus. The expression amounts of FPS in different organs were different significantly (P < 0.05).</p><p><b>CONCLUSION</b>The FPS gene of E. senticosus was successfully cloned for the first time, and provided a stable foundation for studying on its effect and expression control on E. senticosus saponins biosynthesis.</p>

Effect of endophytic fungi on expression amount of key enzyme genes in saponins biosynthesis and Eleutherococcus senticosus saponins content / 中国中药杂志

<p><b>OBJECTIVE</b>To analyze the effect of endophytic fungi on expression amount of key enzyme genes SS (squalene synthase gene), SE (squalene epoxidase gene) and bAS (beta-amyrin synthase gene) in saponin biosynthesis and saponins content in Eleutherococcus senticosus.</p><p><b>METHOD</b>Wound method was used for back meeting the endophytic fungi to E. senticosus. With GAPDH as internal control gene, the expression of key enzyme genes was detected by real time PCR method. E. senticosus saponins content was measured by spectrophotometry method.</p><p><b>RESULT</b>When wound method back meeting P116-1a and P116-1b after 30 d, the expression content of SS improved significantly (P < 0.05), however the back meeting of P109-4 and P312-1 didnt change the expression of SS. After that SS expression showed reduction-equality-reduction varying trend. Thirty days after back meeting P312-1, the expression content of SE improved significantly (P < 0.05). Ninty days after back meeting P116-1b and P312-1, the expression content of SE improved significantly to 130%,161%, respectively (P < 0.05). After 120 d, back meeting four endophytic fungi, the expression of SE were significantly higher than the control (P < 0.05). Back meeting four endophytic fungi form 60 d to 120 d, the expression of bAS was significantly higher than the control (P < 0.05). The back meeting four endophytic fungi improved E. senticosus saponins content significantly (P < 0.05).</p><p><b>CONCLUSION</b>Endophytic fungi P116-1a, P116-1b, P1094 and P312-1 significantly effected the expression of key enzyme genes SS, SE and bAS and then affected E. senticosus saponins content. Among the genes, bAS was key target gene.</p>

Cloning of Eleutherococcus senticosus calmodulin gene and effect of endophytic fungus on expression amount of gene / 中国中药杂志

<p><b>OBJECTIVE</b>To clone calmodulin (CaM) gene in Eleutherococcus senticosus, and study the effect of endophytic fungi on expression amount of CaM gene.</p><p><b>METHOD</b>The CaM full length cDNA sequence was cloned by rapid amplification of cDNA ends (RACE). The gene was analyzed and corresponding structure and functions were predicted by the bioinformatics methods. The expression amount of CaM gene affected of endophytic fungus P116-1a, P116-1b, P1094 and P312-1 was detected by RT-PCR.</p><p><b>RESULT</b>The full length of CaM cDNA was 856 bp containing an ORF of 450 bp that encoded a protein of 149 amino acids. The homologous of predicted protein was almost 100% with plants like Panax ginseng and Daucus carota. RT-PCR results showed that endophytic fungus improved CaM expression amount significantly (P<0.05). The highest expression amount of CaM occurred 90 d after reinoculated with endophytic fungi P1094, up to 2.96 times of the control.</p><p><b>CONCLUSION</b>The CaM gene of E. senticosus was successfully cloned for the first time. The results demonstrated that endophytic fungus of E. senticosus improved CaM expression amount significantly.</p>

Cloning and sequence analysis on cDNA of squalene epoxidase gene in Eleutherococcus senticosus / 中国中药杂志

<p><b>OBJECTIVE</b>To clone and sequence the cDNA of squalene epoxidase gene in Eleutherococcus senticosus.</p><p><b>METHOD</b>Total RNA of E. senticosus was extracted by the improved isothiocyanate method and reverse transcripted into cDNA. The primers were designed depending on the reported SE cDNA sequences of Panax ginseng. The SE cDNAs in E. senticosus was amplified using RT-PCR strategy.</p><p><b>RESULT</b>Sequencing results showed two different cDNA fragments (SE1, SE2) with 1665, 1629 bp each ORF which encoded 554,542 amino acids, respectively. The identities of nucleotides and amino acids between SE1, SE2 were 91.49%, 92.55%. SE1, SE2 had the highest amino acids similarity to the SE1 of P. notoginseng, 93.45%, 94.87% respectively. SE1, SE2 both had a FAD binding domain. The deduced speculated amino acids of SE1, SE2 each had 2,4 membrane-spanning helices.</p><p><b>CONCLUSION</b>The two SE sequences in E. senticosus were firstly separated and reported, which has made foundation for E. senticosus secondary metabolite engineering researches.</p>

Research on standardized preparation of traditional Chinese medicine (III): difference of extracting quantity of anthraquinones from mixed decoction of rhubarb with TCMs containing different ingredients / 中国中药杂志

<p><b>OBJECTIVE</b>To detect the influence of compatibility of rhubarb with different traditional Chinese medicines (TCM) on extracted quantities of AQs, and to provide scientific basis for the clinical code for rhubarb preparation.</p><p><b>METHOD</b>The influence of compatibility of rhubarb with different traditional Chinese medicines (saponin, alkaloids, flavonoids TCMs, animal medicines and mineral medicines) on decocting volume of anthraquinone substance was detected using ultra performance liquid chromatography.</p><p><b>RESULT</b>In comparable conditions, more AQs were extracted from mixed decoction of rhubarb and saponin medicinal materials (Astragali Radix, Notoginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, Polygalae Radix, Ginseng Radix et Rhizoma) than single decocting of rhubarb. The mixed decoction of rhubarb and alkaloid medicinal materials (Coptidis Rhizoma, Sophorae Flavescentis Radix, Prepared Aconiti Lateralis Radix Praeparata, Phellodendri Chinensis Cortex, Aconiti Lateralis Radix) caused a remarkable decrease in extracted quantities of AQs. And the mixed decoction of rhubarb and mineral medicines (Natrii Sulfas, Gypsum Fibrosum, Ostreae Concha, Alumen) also resulted in less extracted quantities of AQs to varying degrees. Besides, more rhubarb AQs were extracted from mixed decoction with Curcuma than single decoction. But less rhubarb AQs were observed in mixed decoction with Lonicerae Flos, Rehmanniae, Artemisiae Herb and Forsythiae Fructus than single decoction to varying degrees. In the study, the maximum extracted quantities of AQs is 2. 3-fold higher than the minimum, the largest difference existed in the extracted quantity of physcion which was 13.5 times.</p><p><b>CONCLUSION</b>In compatibility between rhubarb and different TMCs, mixed decoction and single decoction show different influences on extracted quantity of rhubarb AQs. It is proved that more AQs may be extracted from mixed decoction between rhubarb and saponin medicinal materials, whereas less AQs may be observed in mixed decoction between rhubarb and alkaloid medicinal materials.</p>

COLD or HOT natural attribute of Zuojinwan and Fanzuojinwan based on temperature tropism of mice / 药学学报

This study is to investigate the authenticity between COLD and HOT natural attribute in the famous Chinese medicine formulas--Zuojinwan (Coptis-Evodia 6 : 1) and Fanzuojinwan (Coptis-Evodia 1 : 6) based on mice temperature tropism, and establish an objective method to estimate the difference of two natural attribute by using a cold/hot plate differentiating technology. The results indicated that the COLD nature Zuojinwan could decrease significantly the remaining rate of HOT-symptom rat on warm pad (P < 0.05). That was not notable to COLD-symptom rat. The interference result of COLD-HOT temperature tropism to COLD/HOT symptom rat in Fanzuojinwan was the reverse with the COLD nature Zuojinwan. Meanwhile, biochemical indicators which are relative to energy metabolism such as ATPase enzyme activity and total anti-oxidant capability (T-AOC), had corresponding change in the organism. In the study, the COLD and HOT natural tendency in Zuojinwan and Fanzuojinwan which were composed by the same herbs with different proportion could be expressed qualitatively, quantitatively, objectively and directly with applying animal temperature tropism, and be verified to philosophical idea of treating disease theory with "expelling heat with cold herbs and cryopathy requiring warm prescription", not "expelling heat with heat herbs and cryopathy requiring cold prescription" in ancient traditional Chinese medicine, which brings a new approach in investigation of the nature theory of traditional Chinese medicine.