Exploring a novel class tryptophan hydroxylase 1 inhibitor derived from Sambucus williamsii Hance for the osteoporosis treatment

Objective:Gut-derived serotonin strongly inhibits bone formation by inhibiting osteoblast proliferation.Our previous study demonstrated that the lignan-rich fraction prepared from Sambucus willimasii Hance,a folk herbal medicine used to treat bone fractures and joint diseases in China,exerted bone-protective effects,and its actions were modulated by suppressing the synthesis of gut-derived serotonin via the inhibition of intestinal tryptophan hydroxylase 1(TPH-1).However,there is no direct evidence for the action of lignans on TPH-1.This study aimed to verify the direct action of lignans on the TPH-1 and its influence on serotonin synthesis and bone properties.Methods:Molecular docking and surface plasmon resonance were performed to determine the affinities of lignans to TPH-1.The cell viability and the protein activity and expression of TPH-1 were measured in RBL2H3 cells.The serum serotonin level and bone mineral density upon lignan treatment in ovariectomized mice were determined.Result:The lignans showed high binding scores and binding affinities to TPH-1,inhibited the activity and protein expression of TPH-1,suppressed the serum serotonin levels in ovariectomized mice as well as promoted bone mineral density.Conclusion:This is the first study to report that lignans are novel TPH-1 inhibitors and that these lignans could be potential agents for the management of serotonin-related diseases,including osteoporosis.

Effects of Abrus cantoniensis Hance Extract on Blood Lipid of Laying Hen Feed with High Energy and Low Protein Diet

[Objectives]To explore the effects of Abrus cantoniensis Hance(ACH)extract on blood lipid indicators of laying hen fed with high energy and low protein diet.[Methods]Sixty 90-day-old laying hens were randomly divided into five groups:the blank control group(basic diet),the model group(high-energy and low-protein diet,HELPD),the low-dose group(HELPD+0.5 g ACH extract per hen,LACH),and the medium-dose group(HELPD+1 g ACH extract per hen,MACH),high dose group(HELPD+2 g ACH extract per hen,HACH).The ACH extract was administrated by drinking water for 48 d.[Results]Different doses of ACH could improve the pathological changes induced by high energy and low protein.ACH extract had no significant effect on blood routine indicators of laying hens(P>0.05).The contents of total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)in the model group were significant-ly higher than those in the control group(P<0.05),while the content of high density lipoprotein cholesterol(HDL-C)was significantly lower than that in the control group(P<0.05).There was no significant difference in blood lipid between LACH group and model group(P>0.05).In MACH and HACH groups,the contents of TC,TG and LDL-C were significantly lower than those in the model group(P<0.05),and the content of HDL-C was significantly higher than that in the model group(P<0.05).[Conclusions]The ACH extract can regulate theHELPD-induced dyslipidemia in laying hens.

Construction of Prokaryotic Expression Vectors of EBP1 Gene from Nervilia Fordii (Hance) Schltr.

[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.

Optimization of the Method to Obtain Effective Sterile Explants in Tissue Culture of Ardisia mamillata Hance

In the present study, we optimized the procedures to pretreat the parental material, to collect, sterilize and inoculate the explants in the tissue culture of Ar- disia mamillata Hance, so that more effective sterile explants can be obtained. The results revealed the optimal procedures. In detail, the parental materials were trans- ferred and pretreated in laboratory in February. The stem tips of new branches were collected in middle March, cleaned with eradicator and then rinsed with tap water for 1-2 h. After that, the explants were surface sterilized with 70% alcohol for 30 s, rinsed with sterile water 2-3 times, sterilized with 0.1% mercuric chloride for 4 min, rinsed with sterile water three times and sterilized with 0.1% mercuric chlo- ride for 4 min again. Finally, they were inoculated into medium after being rinsed with sterile water 5-8 times. By this method, the pollution rate of explants can be greatly reduced while their survival rate can be significantly improved.

Zn Accumulation and Subcellular Distribution in the Zn Hyperaccumulator Sedurn alfredii Hance

Zn accumulation and subcellular distribution in leaves of the hyperaccumulating ecotype （HE） and non-hyperaccumulating ecotype （NHE） of Sedum alfredii Hance were studied using radiotracer and gradient centrifugation techniques. Leaf Zn accumulation in the HE of S. alfredii was 18.5-26.7 times greater than that in the NHE when the plants were grown at 1-500μmol Zn L-1. Leaf section uptake of 65Zn was highly dependent on external Zn levels. Greater 65Zn uptake in HE was noted only at external Zn levels 〉 100μmol L-1. Zinc subcellular distribution in the leaves of the two ecotypes of S. alfredii was： cell wall 〉 soluble fraction 〉 cell organelle. However, more Zn was distributed to the leaf cell wall and soluble fractions for HE than for NHE. In the leaf of HE, 91%-94% of the Zn was found in the cell walls and the soluble fraction and only 6%-9% Zn was distributed in the cell organelle fraction. For NHE, about 20%-26% Zn was recovered in the cell organelle fraction. In stems, Zn distribution to the ceil wail fraction was approximately two fold greater in the HE than that in the NHE. For the hyperaccumulating ecotype of S. alfredii, the cell wall and the vacuole played a very important role in Zn tolerance and hyperaccumulation.

Effect of Alpinia officinarum Hance alcohol extracts on primary dysmenorrheal

Objective:To study the effect of Alpinia officinarum Hance(A.officinarum) 80% alcohol extract on the primary dysmenorrhea.Methods:A.officinarum 80% alcohol extract were enriched by macroporous adsorption resins.Female mice of primary dysmenorrhea model were established by oxytocin induction; the effects of A.officinarum 80% alcohol extract on primary dysmenorrhea were observed by body twist method; and the homogenate level of prostaglandin F_(2α)(PGF_(2α)),prostaglandin E_2(PGE_2) and Ca^(2+) in the uterus were observed in oxytocin-induced female mice.Results:The writhing frequency of primary dysmenorrhea mice was significantly decreased after treatment of A.officinarum 80% alcohol extract and the level of PGF_(2α),PGE_2 and Ca^(2+) in mice uterus was significantly decreased(P<0.05,P<0.01) in groups of mice treated with middle and high dosage of A.officinarum 80% alcohol extract compared with that of model group.Conclusions:These findings suggest that A.Officinarum 80% alcohol extract can significantly relieve primary dysmenorrhea.

Two New Phloroglucinol Glycosides from Lysidice rhodostega Hance

Two new phloroglucinol glycosides, lysidiciside A (1) and lysidiciside B (2), were isolated from the roots of Lysidice rhodostega Hance. Their structures were determined as 1-[(3-methylbutyryl)phloroglucinol]-β-D- glucopyranoside (1), 1- [(3-methylbutyryl)phloro- glucinol]-β-D-glucopyranosyl-5-O-β-D-glucopyranoside (2) on the basis of chemical and spectral analysis .

A novel diarylheptanoid bearing flavonol moiety from the rhizomes of Alpinia officinarum Hance

A novel diarylheptanoid bearing flavonol moiety,named officinin A(1),along with two known compounds galangin and kaempferide were isolated from the rhizomes of Alpina officinarum Hance.The structure elucidation was accomplished by HRESI -MS,1D and 2D NMR methods.

Zinc adsorption and desorption characteristics in root cell wall involving zinc hyperaccumulation in Sedum alfredii Hance

Radiotracer techniques were employed to characterize 65Zn adsorption and desorption in root-cell-wall of hyperac-cumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end of a 30 min short time radioisotope loading period, comparable amounts of 65Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more 65Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of 65Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more 65Zn accumulated in desorbed root-cell-wall of NHE. Zn2+ binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn2+ concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn2+, whereas the desorption characteristics were different, and with most of 65Zn binding on root of HE being available for loading into the xylem, as a result, more 65Zn was translocated to the shoot.

A new homoflavonoid from the seed of Caesalpinia minax Hance

A new homoflavonoid was isolated from the seed of Caesalpinia minax Hance.On the basis of spectral evidences,its structure was established and named as minaxin.

Integrated Fertilization Regimes Boost Heavy Metals Accumulation and Biomass of Sedum alfredii Hance

The hyperaccumulator Sedum alfredii Hance(S.alfredii)may be employed for zinc(Zn)and cadmium(Cd)-polluted soil remediation.However,the low phytoremediation efficiency,related to the low biomass production,limits its use with that purpose.In this experiment,nitrogen(N),phosphorus(P),and potassium(K)fertilizers,and organic manure were applied to investigate the phytoremediation ability of S.alfredii.Hydroponic and pot experiments were conducted using Zn-Cd polluted soil.The hydroponic experiment indicated that appropriate fertilizer application could increase(p<0.05)the amount of accumulated Zn and Cd in S.alfredii.When N supply ranged from 0.5 to 2.5 mmol L^(−1),it could improve growth and accumulation of Zn and Cd in whole plants of S.alfredii.The 1 mmol L^(−1) N was an optimal N dosage for shoot biomass production and Cd accumulation in shoots,while the 2.5 mmol L^(−1) was an optimal N dosage for Zn accumulation in shoots.Both low(<0.05 mmol L^(−1))and high(>0.8 mmol L^(−1))P supply decreased growth,and Zn/Cd accumulation in whole plants of the studied species.The 0.1 mmol L^(−1) P was an optimal dosage for S.alfredii biomass production and Zn/Cd accumulation in shoots.The supply levels within the range from 0.3 to 1 mmol L^(−1) K could significantly improve the biomass production of S.alfredii and its capability to accumulate Zn and Cd in the biomass.The 0.5 mmol L^(−1) K was an optimal dosage for the whole biomass production and Zn accumulation in shoots,while the 1 mmol L^(−1) was an optimal K dosage for Zn accumulation in shoots,which was 17.2%higher than the control.Moreover,the soil pot experiment showed that the combination of organic(fermented manure)and inorganic fertilizers made significant effects on the Zn and Cd-polluted soil remediation by S.alfredii.These effects varied,however,with the application of different proportions of N,P,K and organic matter.The Zn accumulation by S.alfredii reached the highest efficiency ability under the highest fertilizer mixing rate(N:50 mg kg^(−1),P:40 mg kg^(−1),K:100 mg kg^(−1),organic matter:1%).Even more,S.alfredii showed the strongest ability to accumulate Cd with a lower fertilizer mixing rate(N:25mg kg^(−1),P:20mg kg^(−1),K:50 mg kg^(−1),organic matter:0.5%).

Differential generation of hydrogen peroxide upon exposure to zinc and cadmium in the hyperaccumulating plant specie(Sedum alfredii Hance)

Sedum alfredii Hance has been identified as zinc(Zn) and cadmium(Cd) co-hyperaccumulator.In this paper the relationships of Zn or Cd hyperaccumulation to the generation and the role of H2O2 in Sedum alfredii H.were examined.The results show that Zn and Cd contents in the shoots of Sedum alfredii H.treated with 1000 μmol/L Zn2+ and/or 200 μmol/L Cd2+ increased linearly within 15 d.Contents of total S,glutathione(GSH) and H2O2 in shoots also increased within 15 d,and then decreased.Total S and GSH contents in shoots were higher under Cd2+ treatment than under Zn2+ treatment.However,reverse trends of H2O2 content in shoots were obtained,in which much higher H2O2 content was observed in Zn2+-treated shoots than in Cd2+-treated shoots.Similarly,the microscopic imaging of H2O2 accumulation in leaves using H2O2 probe technique showed that much higher H2O2 accumulation was observed in the Zn2+-treated leaf than in the Cd2+-treated one.These results suggest that there are different responses in the generation of H2O2 upon exposure to Zn2+ and Cd2+ for the hyperaccumulator Sedum alfredii H.And this is the first report that the generation of H2O2 may play an important role in Zn hyperaccumulation in the leaves.Our results also imply that GSH may play an important role in the detoxification of dissociated Zn/Cd and the generation of H2O2.

A New Flavonol Glycoside from Anaphalis sinica Hance

A new flavonol glycoside, 5,7,3,4 tetrahydroxy 3 methoxyflavonol 3 O β D glucopyranoside(1) was isolated along with quercetin(2), quercetin 3 O α L rhamnopyranoside(3), quercetin 3 O β D glucopyranoside(4) and kaempferol 3 O β D (6' O p coumaroyl) glucopyranoside(5) from Anaphalis sinica Hance. Their structures were determined on the basis of spectral data(MS, UV, IR, 1H and 13 C NMR). The structure of compound 1 was confirmed by X ray crystallographic analysis.

Effect of Light Intensity on Leaf Photosynthetic Characteristics and Accumulation of Flavonoids in Lithocarpus litseifolius (Hance) Chun. (Fagaceae)

The active compounds in herb drugs are mainly secondary metabolites, which are greatly influenced by external conditions. Particularly, light intensity has a great influence on the photosynthesis and accumulation of secondary metabolites. In this study, the light intensity was changed, and the influence of the light intensity on leaf photosynthetic characteristics, antioxidant enzyme activity and flavone contents of Lithocarpus litseifoliusp (Hance) Chun. was discussed. The results showed that (1) L. litseifolius is a typical heliophyte. As the light intensity decreased, the contents of chlorophyll a (Chl a), chlorophyll b (Chl b) and total chlorophyll (Chl a + b) all increased. However, the Chl a/b ratio gradually decreased. The daily variation of net photosynthetic rate changed from a double-peak curve under natural light to single-peak curve under 20% full light two shading nets. (2) As the light intensity decreased, the SOD and POD activities and the soluble proteins content decreased in the leaves. In contrast, the malondialdehyde content increased. (3) The flavone content was the highest in leaves at one shading nets 60% of full light, and the lowest occurred in leaves at two shading nets 20% of full light. A comprehensive analysis indicated that an appropriate shading could induce shade tolerance in the leaves of L. litseifolius, which promoted the accumulation of flavonoids. L. litseifolius should be planted in the lower position of sunny slope or the upper position of shady slope to increase the content of flavone in the leaves.

Alpinia officinarum Hance extract alleviates particulate matter-induced lung injury in mice

Objective: To evaluate the effect of Alpinia officinarum Hance(A. officinarum) extract on lung injury caused by particulate matter(PM). Methods: The Kunming mice were intranasally instilled with PM and treated with A. officinarum extract for 3 weeks. Bronchoalveolar lavage fluid, blood and lung samples were collected for biochemical, serological and histopathological studies. Results: Serological analysis showed that albumin levels, lactate dehydrogenase and alkaline phosphatase activities in bronchoalveolar lavage fluid were significantly reduced after administrations of 50, 100 and 200 mg/kg of A. officinarum extracts to the PM injured mice. Markers of oxidative stress, nitric oxide, malondialdehyde levels and nitric oxide synthase activities, were significantly decreased. Correspondingly, total superoxide dismutase activity was improved dramatically. The expressions of interleukin-6 and tumor necrosis factor alpha were also down-regulated obviously. In addition, pathological sections of lung tissue showed that A. officinarum could reduce the infiltration of inflammatory cells, pulmonary edema and pulmonary fibrosis. These results showed that A. officinarum extract could alleviate PMinduced lung injury via reducing the permeability of cell membranes in lung tissue, eliminating oxidative stress and relieving inflammatory response.Conclusions: A. officinarum extract was an efficient treatment for PM-induced lung injury in mice, and it may be a promising therapeutic agent in future.

Acute Toxicity Test of Abrus cantoniensis Hance Extract

[Objectives]To investigate the acute toxicity of petroleum ether extract,n-butanol extract,ethyl acetate extract and water extract to mice.[Methods]The medium lethal dose(LD50)in mice could not be determined,so the maximum dose method was used for experiment.Mice were not fed but allowed to drink water for 16 h before the experiment.There were 74 SPF Kunming Mice,half male and half female,and it was divided into extract group and blank control group.The toxicity and death were recorded after intragastric administration and regular close observation.If any mice died,the dead mice were dissected.The changes of heart,liver,spleen,lung,kidney,brain,stomach,small intestine and other organs were observed with the naked eye.At the end of the 14 d observation period,the surviving mice in each group were killed and dissected.The gross pathological changes of the main organs were observed with the naked eye according to the same method mentioned above.[Results]There were no obvious toxic symptoms and no death in the mice.[Conclusions]The maximum doses of petroleum ether extract,n-butanol extract,ethyl acetate extract and water extract in mice were 1.61,1.71,1.65 and 1.79 g/L,respectively.Thus it was found that the quarterly tests of petroleum ether extract,n-butanol extract,ethyl acetate extract and water extract suggested that the medicine was safe.

Organic Materials Could Improve the Phytoremediation Efficiency of Soil Potentially Hazardous Metal by Sedum alfredii Hance

Soil potentially hazardous metal(PHM)is continually attracting public attention worldwide,due to its highly toxic properties and potentially huge damage to human being through food chain.Phytoremediation is an effective and eco-friendly way in remediation technology.A pot experiment was carried out to investigate the effect of different organic materials(biogas residue(BR),mushroom residue(MR),and bamboo-shoot shell(BS))application on phytoremediation of two PHM-contaminated soils(Fuyang soil as‘heavily-polluted soil’and Wenzhou soil as‘moderately-polluted soil’,respectively)by Sedum alfrecdii Hance.The results indicated:1)for moderately-polluted soil,the 5%BR treatment had the strongest activation to Cu and Zn,for heavily-polluted soil,1%BS treatment had the highest activation effect for Cu,Zn,Pb and Cd.2)the above-ground biomass of Sedum alfredii Hance increased with the addition rate of organic materials.3)for Cd uptake of Sedum alfredii Hance in moderately-polluted soil,only 1%BS treatment had a better accumulation effect,compared to the control,for Zn element,MR treatments were weaker than the control,while other treatments were better than the control,of which 5%BR,1%BS and 5%BS accumulated more Zn element by 39.6%,32.6%and 23.8%,respectively;in heavily-polluted soil,the treatments of 5%BS,1%BR and 5%BR accumulated more Cd than the control by 12.9%,12.8%and 6.2%,respectively,the treatments with organic materials addition promoted Zn accumulation in shoots of Sedum alfredii Hance,and the best treatment was 5%BS.Therefore,an appropriate application rate of BS and BR could improve the remediation efficiency for Zn/Cd contaminated soils by Sedum alfredii Hance.

Effects of Interaction of Soil Moisture and Organic Matter on Powdery Mildew Disease and Growth of Heracleum moellendorffii Hance

Taking annual Heracl'eum moellendorffii Hance as the material,this research studied the epidemic dynamics of H.moellendorffii Hance powdery mildew and plant growth by using the interactive methods of soil moisture and soil organic matter.The results showed that the interaction of higher soil moisture content(60%-80%)and soil organic matter content(75-100 g•kg^(-1))had few diseases,dosease index and area under disease progression curve(AUDPC)compared with those of other treatments,thus could effectivley control powdery mildew disease.Moreover,higher soil moisture and organic matter content increased the yield,contents of soluble sugar,soluble protein,vitamin C and coumarin in H.moellendorffii Hance.

Qualitative Identification and Content Determination of Psoralen in Ficus Pandurata Hance

[Objectives]This study aimed to establish a qualitative identification and content determination method for psoralen in Ficus pandurata Hance and compare the psoralen contents in roots,stems and leaves of F.pandurate Hance and Ficus pandurata Hance var.holophylla Migo.[Methods]Thin-layer chromatography(TLC)was used for qualitative identification,and the content of psoralen was determined by high-performance liquid chromatography.The chromatographic conditions were as follows:column,Agilent ZORBAX Eclipse Plus C18(250 mm×4.6 mm,5μm);mobile phase,methanol-water(55∶45);flow rate,1 mL/min;detection wavelength,246 nm;column temperature,30℃;and injection volume,10μL.[Results]The TLC chromatogram of F.pandurate Hance showed clear spots and good separation.The concentration of psoralen detected in the range of 2.06-41.20μg/mL had a good linear relationship with the peak area(r=0.9999).The RSD values of the precision,stability and reproducibility tests were all less than 2%.The average recovery rate was 100.2%(RSD=1.13%,n=6).[Conclusions]The established method is simple,easy,accurate and reproducible.It can be used for quality control of F.pandurate Hance.

Responses of Nitrogen Metabolism to Rhizobium Inoculation in Abrus mollis Hance

In different periods, the contents of nitrogen, ammonia nitrogen and nitrate nitrogen in different parts of Abrus mollis Hance were determined for each period by nesslerization, ninhydrin method and salicylic acid method in pot experiment. By using related data statistical software, the effects of Rhizobium inoculation on nitrogen metabolism, accumulation and distribution in Abrus mollis Hance were analyzed preliminarily.