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Objective To isolate the secondary metabolites of endophytic fungus Aspergillus sp. J218 from Anectochilus roxburhii. Methods Different chromatographic methods, including Sephadex LH-20 and silica gel chromatography as well as HPLC, were used to isolate compounds from the EtOAc fraction of the solid fermentation of J218, and their structures were identified by spectral methods. Results Ten compounds were isolated from the fermentation of J218 and their structures were individually identified as kotanin（1）, flavasperone（2）, aurasperone B（3）, fonsecinone B（4）, fonsecinone D（5）, ensidol A（6）, fonsecinone A（7）, fonsecinone C（8）, aurasperone A（9）, and fonsecinone F（10）. Conclusion Most compounds isolated from endophytic fungus Aspergillus sp. J218 in Anectochilus roxburhii were identified as dimeric naphthopyrones. These results suggest that this strain contains rich dimerization synthase, which could provide clues for the further exploration of the rational biosynthesis pathway of dimeric naphthopyrones in this strain.
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Objective To perform the pharmacognostic identification of Anoectochilus lylei and establish the foundation for its accurate identification and further development. Methods The macroscopic identification and microscopic identification methods were used to identify A. lylei. Results A. lylei has ovate leaf shape, possessing red reticulated veins. Inverted flowers have Y-shaped and white lip. The anterior part of lip is two-lobed, and the lobes are linear-oblong. There are 1 to 3 shorts serrations on each side of the middle part of lip. Microscopic characteristics mainly show as follows: the cortex is broad in the transverse section of roots and stems; 1-5 and 1-7 vascular bundles in the xylem of transverse section of roots and stem, respectively. Collateral vascular bundle in the main veins of transverse section of leaves. There are multitudinous types of stomas in the leaf abaxial epidermis, most of which are anomocytic. Conclusion These characteristics could provide reference for the correct identification of A. lylei.
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Objective To study the effects of endophytic fungus Epichloë bromicola SH09 on the plant growth and accumulation of active components in Salvia miltiorrhiza, and improve the quality of medicinal plant S. miltiorrhiza. Methods E. bromicola SH09 solid bacterial fertilizer was prepared and co-cultured with S. miltiorrhiza for 60 d and 120 d. Four morphological indexes, fresh weight of roots, dry weight of roots, and the contents of four tanshinones and two phenolic acids in the roots of S. miltiorrhiza from treated group and control group were assayed, respectively. Results After 60 d and 120 d co-culture, E. Bromicola SH09 significantly increased the tiller number, plant height, leaf number, leaf area, fresh weight of roots, dry weight of roots, and the content of tanshinones and phenolic acids in S. miltiorrhiz. Conclusion The endophytic fungus E. bromicola SH09 can effectively promote the plant growth and improve the accumulation of active components in S. miltiorrhiza, which not only broadens the new ecological functions of endophytic fungi, but also improves the quality of medicinal plant S. miltiorrhiza.
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Objective To investigate the change of biological characteristics after stable knockdown of the coding gene of 3-methylcrotonyl-coenzyme A carboxylase β subunit (MCCC2) expression in DU145 by lentivirus shRNA. Methods Three groups were included in this study. shNC was the control group in which MCCC2 was negatively knocked down in DU145. shMCCC2 was the experimental group in which MCCC2 was knocked down. DU145 was the blank group without any treatment. The expression of MCCC2 was assessed by Western blot and qPCR. The proliferation of DU145 cells was detected by CCK8 assay. The migration ability of DU145 was detected by transwell. The apoptosis of DU145 cells was detected by flow cytometry. Results The expression level of MCCC2 in shMCCC2 group was significantly lower than that in shNC group (0.22 ± 0.02 vs 0.61 ± 0.06, P < 0.001). The proliferation (2.24 ± 0.04 vs 3.13 ± 0.15) and migration (23.96 ± 1.85 vs 49.73 ± 0.63) of DU145 cells in shMCCC2 group was significantly lower than that in shNC group, whereas the apoptosis (12.64 ± 0.30 vs 3.68 ± 0.02) of DU145 cells in shMCCC2 was significantly higher than that in shNC group. Conclusion MCCC2 knockdown significantly inhibited the proliferation and migration, and induced apoptosis of DU145 cells, which indicated that the down-regulation of MCCC2 is correlated with the change of tumor biological characteristics of DU145 cell line and can be a potential target for the treatment of prostate cancer.
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Objective To identify the crude drugs of Anoectochilus burmannicus, and clarify its original plant pharmacognostical and microscopic characteristics. Methods The pharmacognostical identification method was used to observe the original plant, tissue structure and microscopic characteristics of A. burmannicus. Results Leaves were ovate or ovate elliptic with golden-red veins. Non-inverted yellow flowers had Y-shaped and yellow labellum, which were anteriorly enlarged and 2-lobed. The lobes were narrowly oblong or narrowly oblanceolate. The middle part of labellum was narrow to form a 10 mm long structure with margin narrowly winged. In the microscopic structure, the cortex is obvious in the cross sections of root and stem, together with needle crystals of calcium oxalate and mucous cells. The upper epidermal cells on the cross section of the leaves were papilloid in shape, whereas diverse stomas existed among the lower epidermal cells, with anomocytic stomas as the major type. Needle crystals of calcium oxalate and conduits can be found in the powder. Conclusion These data provide a reference for the identification and resource development and utilization of A. burmannicus.
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Objective To study the influence of eleutheroside E (ELE)on the learning and memory abilities of mice in-duced by para-chlorophenylalanine (PCPA) and on hippocampus′monoamine neurotransmitters of serotonin(5-HT) ,5-hydrox-indole acetic acid (5-HIAA) ,dopamine (DA) and norepinephrine (NA) .Methods Balb/c mice were randomized into control group ,model group ,positive group and three treatment groups (high ,moderate and low dose group) .The mouse injury model was established by PCPA abdominal injection .The mice′s abilities of learning and memory were detected in the Morris water maze test after 14 days of prevention medicine .The contents of 5-HT ,5-HIAA ,DA and NA in mice′s hippocampus were de-tected by Elisa kit .Results The behavior results showed that ,compared with control group ,the times across the platform by model group decreased significantly (P<0 .01) .The times across the platform by high and moderate dose groups increased sig-nificantly (P<0 .01) after the administration of ELE .The contents of 5-HT and 5-HIAA of the model group were lower than those of the control group (P<0 .01) .The contents of NA was lower too (P<0 .05) .The contents of 5-HT and 5-HIAA of different dose groups were much higher than those of the model group .Conclusion The enhancement of learning and memory ability in the mouse injury model established by PCPA might be related with the improvement of the contents of 5-HT ,5-HIAA ,DA ,NA in mice′s hippocampus by ELE .
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Objective To optimize the extraction technology of total falvonoids from Hypericum sampsonii .Methods On the basis of single factor test ,solid‐liquid ratio ,ethanol concentration ,extraction time ,extraction temperature and extrac‐tion times on the extraction rate were studied by taking the content of total falvonoids as indexes .Orthogonal test was conduc‐ted to establish the extraction technology of total falvonoids from Hypericum sampsonii .Results The influence degree of fac‐tors on the extraction rate of total falvonoids was in the order of ethanol concentration ,solid‐liquid ratio ,extraction times ,and temperature .The optimum extraction condition was confirmed as follows:ethanol concentration of 70% ,extraction tempera‐ture of 60 ℃ ,and extraction times of 3 × 1 .5 h ,solid to liquid ratio of 1:20 .Under this condition ,the average extraction rate of total falvonoids reached 2 .38% .Conclusion The test was reasonable in design and credible in findings ,and was easily oper‐ated ,which could provide scientific basis for the batch extraction of total flavonoids from Hypericum sampsonii .