Effect of Milk Processed Arnebiae Radix on Body Temperature of Fever Rats

[Objectives]To investigate the synergistic effect of Arnebiae Radix after processing.[Methods]The effects of raw Arnebiae Radix and milk processed Arnebiae Radix on hypothermia in yeast-induced febrile rats were compared.[Results]The processed and unprocessed Arnebiae Radix at high,medium and low doses all had a certain effect on inhibiting the rise of body temperature in rats.The high dose unprocessed group,the medium dose processed group and the high dose processed group had the best inhibitory effect on body temperature,the low dose processed group could delay the fever time,and the low dose unprocessed group had poor inhibitory effect on fever.[Conclusions]The prepared Arnebiae Radix has enhanced drug effect,and milk processed Arnebiae Radix can be used to replace common Arnebiae Radix to reduce the dosage of Arnebiae Radix and save Arnebiae Radix resources.

Qualitative and quantitative analysis of Arnebiae Radix and Dictamni Cortex and efficacy study of herbal extracts on allergic contact dermatitis using 3D human reconstructed epidermis

Objective:To evaluate the quality of Arnebiae Radix(AR)and Dictamni Cortex(DC)and study the efficacy of herbal extracts of these two herbs on the treatment of allergic contact dermatitis(ACD).Methods:Qualitative and quantitative analysis of effective components was performed using High Performance Thin Layer Chromatography(HPTLC),High Performance Liquid Chromatography(HPLC),and HPLC-Quadrupole Time of Flight-Mass Spectrometry(HPLC-QTOF-MS).In vitro allergic ACD 3D model was established by incubating 3D reconstructed human epidermis(RHE)with skin sensitizer,potassium dichromate.A total of 65 gene expression that were associated with ACD,which included 24 antioxidant responsive element(ARE)and 41 SENS-IS genes were quantified by q RT-PCR.More than or equal to 10 ARE genes and 18 SENN-IS genes were induced by 1.3-fold,demonstrating the successful establishment of in vitro ACD model.Oil extracts of AR and DC were applied on the in vitro ACD model to study the efficacy.Results:Batch 3 of AR and batch 2 of DC showed presence of all active ingredients with the highest concentrations.Active ingredients of the herbs were extracted using a special oil and formulated into herbal oil extracts.The herbal oil extracts were able to down regulate the induced genes in the in-vitro ACD skin model,bringing the tissue back to homeostatic status.Conclusion:The oil extracts showed the potent efficacy of using AR and DC in ACD treatment.The combination study will be done to optimize the formulation ratio which will be developed into a topical cream.

栽培条件对新疆紫草生长发育的影响

采用田间与室内试验观察方法,研究了新疆紫草生长发育特点.得出初步结果:本种适宜在气候凉爽、湿润、光照好、土壤弱酸至弱碱性、土质疏松的环境下生长;适宜温度在9～22℃之间;光照条件差,则生长发育缓慢;性喜湿,不耐涝,人工栽培不宜用渠灌.此外,新疆紫草为多年生草本,生长发育大致可分为苗期、莲座期、抽茎期、花期、果期.栽培的实生苗第一次开花约需2～3年.

Therapeutic effect of a hydroxynaphthoquinone fraction on dextran sulfate sodium-induced ulcerative colitis

AIM:To evaluate the therapeutic effect of hydroxynaphthoquinone mixture(HM) on dextran sulfate sodium(DSS)-induced colitis and explore the underlying mechanisms.METHODS:BALB/c mice received 3.5% DSS for 6 d to induce ulcerative colitis.Groups of mice were orally administered HM 3.5,7 and 14 mg/kg and mesalazine 200 mg/kg per day for 7 d.During the experiment,clinical signs and body weight,stool consistency and visible fecal blood were monitored and recorded daily.A disease activity index score was calculated for each animal.At the conclusion of the experiment,the colonic histopathological lesions were evaluated.Myeloperoxidase(MPO) activity and tumor necrosis factor-α(TNF-α) levels were determined.Protein expression levels of TNF-α,nuclear factor-κB(NF-κB) p65,inhibitor of κB(IκB) and phosphorylation of IκB(p-IκB) were analyzed by Western blot analysis.RESULTS:Administration of 3.5% DSS for 6 d successfully induced acute colitis associated with soft stool,diarrhea,rectal bleeding,and colon shortening,as well as a loss of body weight.Administration of HM effectively attenuated the severity of colonic mucosa injury.For histopathological analysis,HM treatment improved histological alterations and lowered pathological scores compared with the DSS only group.This manifested as a reduction in the extent of colon injury and inflammatory cell infiltration,as well as the degree of mucosal destruction.In addition,HM at doses of 7 and 14 mg/kg significantly decreased MPO activity in colonic tissue(0.98 ± 0.22 U/g vs 1.32 ± 0.24 U/g,0.89 ± 0.37 U/g vs 1.32 ± 0.24 U/g tissue,P < 0.05) and serum TNF-α levels(68.78 ± 7.34 ng/L vs 88.98 ± 17.79 ng/L,64.13 ± 14.13 ng/L vs 88.98 ± 17.79 ng/L,P < 0.05).Furthermore,HM down-regulated the expression of TNF-α,NF-κB p65 and p-IκBα in colonic tissue while up-regulating IκBα protein expression.These results suggest that the significant anti-inflammatory effect of HM may be attributable to its inhibition of TNF-α production and NF-κB activation.CONCLUSION:HM had a favorable therapeutic effect on DSS-induced ulcerative colitis,supporting its further development and clinical application in inflammatory bowel disease.

Effectiveness of a hydroxynaphthoquinone fraction from Arnebia euchroma in rats with experimental colitis

AIM:To evaluate the potential effectiveness of hydroxynaphthoquinone mixture(HM)in rats with 2,4,6-trinitrobenzene sulfonic acid(TNBS)-induced colitis.METHODS:Colitis was induced by intracolonic administration of TNBS(80 mg/kg,dissolved in 50%ethanol).Rats were treated daily for 7 d with HM(2.5,5,10 mg/kg)and mesalazine 100 mg/kg 24 h after TNBS instillation.Disease progression was monitored daily by observation of clinical signs and body weight change.At the end of the experiment,macroscopic and histopathologic lesions of rats were scored,and myeloperoxidase(MPO)activity was determined.We also determined inflammatory cytokine tumor necrosis factor(TNF)-αlevel by ELISA,Western blotting and immunochemistry to explore the potential mechanisms of HM.RESULTS:After intracolonic instillation of TNBS,animals developed colitis associated with soft stool,diarrhea and marked colonic destruction.Administration of HM significantly attenuated clinical and histopathologic severity of TNBS-induced colitis in a dose-dependent manner.It abrogated body weight loss,diarrhea and inflammation,decreased macroscopic damage score,and improved histological signs,with a significant reduction of inflammatory infiltration,ulcer size and the severity of goblet cell depletion(all P<0.05 vs TNBS alone group).HM could reduce MPO activity.In addition,it also decreased serum TNF-αlevel and down-regulated TNF-αexpression in colonic tissue.This reduction was statistically significant when the dose of HM was 10 mg/kg(P<0.05 vs TNBS alone group),and the effect was comparable to that of mesalazine and showed no apparent adverse effect.The underlying mechanism may be associated with TNF-αinhibition.CONCLUSION:These findings suggest that HM possesses favourable therapeutic action in TNBS-induced colitis,which provides direct pharmacological evidence for its clinical application.

Radical scavenging and antibacterial activity of Arnebia benthamii methanol extract

Objective:To evaluate in vitro antioxidant and antibacterial activity of methanolic extract of Arnebia benthamii（A.benthamii） whole plant.Methods:Plasmid damage was analyzed by agarose gell electrophoresis.Calf thymus DNA was monitored by TBARS formation.DPPH, reducing power and lipid peroxidation was evaluated by using standard procedures.Antibacterial assay was monitored by disc diffusion method.Results:DPPH radical scavenging and hydroxyl radical scavenging potential of the plant revealed that the extract to be active radical scavenger.Reducing(Fe3+-Fe2+) power and lipid peroxidation inhibition efficiency（TBARS assay） of the extract was also evaluated and the extract showed promising activity in preventing lipid peroxidation and might prevent oxidative damages to biomolecules.The extract offered a significant protection against plasmid and calf thymus UNA damage induced by hydroxyl radicals. The extract was also evaluated on different bacterial strains and the maximum antibacterial activity was exhibited against Escherichia coli（E.coli） when compared with standard drug. Conclusions:These findings demonstrate that the methanol extract of A.benthamii has excellent anti-oxidant activities and could be considered as a potential source of lead molecules for pharmaceutical industries.

Comparative Study on Effects of Arnebia Euchroma (Royle) Johnst Granular and Decoction Forms on Medical AbortionwithMifepristone andMisoprostol

Objective To compare the effects of 2 dosage-forms (granular and decoction) of Arnebia euchroma (royle) Johnst (Arnebia EJ in short below) on medical abortion with that of mifepristone combined with misoprostol Methods Totally 648 women, who had pregnancy of 38-45 d and were willing to terminate pregnancy with mifepristone and misoprostol, were randomly divided into 3 groups, each of which was respectively given granular of Arnebia EJ, placebo granular, or decoction of Arnebia EJ besides mifepristone and misoprostol. The abortion results, bleeding duration, menstruation recovery and side-effects were observed. Results Neither complete abortion rates nor average bleeding durations of the granular group and the decoction group were significantly different (P>0.05). The complete abortion rate and bleeding duration of the two groups were respectively higher and shorter than those of the placebo group (P<0.05). However, the menstruation recov- ery was not significantly different among the three groups (P>0.05). The decoction of Arnebia EJ caused significantly more nausea and vomiting than the other groups (P<0.05). Conclusion The granular form did not have the odor of Arnebia EJ, and caused much less nausea and vomiting compared with the decoction form. The granular and decoc- tion forms were equally effective in improving the results of medical abortion. There- fore it is necessary to conduct further studies on the granular form of Arnebia EJ.

Acetylshikonin Inhibits Colorectal Cancer Growth via PI3K/Akt/mTOR Signaling Pathway

Background: Acetylshikonin, a major constituent isolated from Arnebia euchroma, is a potential candidate for anti-colorectal cancer drugs. However, the potential activity and underlying mechanism of Acetylshikonin against colorectal cancer remain unclear. Methods: In this study, Acetylshikonin was isolated from the active CHCl3 extract of Arnebia euchroma using activity-guided screening, and elucidated by the extensive spectroscopic analysis and comparison with literature data. Human colorectal cancer cells HT29, DLD-1, HCT116 or Caco-2 were exposed to different concentrations of Acetylshikonin (6.25 - 100 μg/mL) for 24 or 48 h. Cell viability, cell apoptosis and cell cycle distribution were detected. The activity of Acetylshikonin and potential mechanism of the phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway were evaluated in vitro and vivo. Results: We found that Acetylshikonin exhibited remarkable anti-proliferative activity in a dose-dependent manner against HT29 cells with the IC50 values of 60.82 μg/ml and 30.78 μg/ml at 24, 48 h, respectively. Moreover, Acetylshikonin induced cell cycle arrest at G0/G1 phase and early apoptosis through inhibition of PI3K/Akt/mTOR pathway. Furthermore, the assays of cell inhibition, early apoptosis and G0/G1 phase distribution showed that suppression of the PI3K/Akt pathway using LY294002 enhanced the anti-cancer effect of Acetylshikonin. Similarly, Acetylshikonin also decreased the growth of tumour in colorectal cancer xenografts in mice through PI3K/Akt/mTOR pathway. Conclusions: To sum up, these new findings provided a framework for further exploration of Acetylshikonin which possessed the potential antitumor activity by inhibiting PI3K/Akt/mTOR pathway.

Enhancement of Shikonin Production in Cell Suspension Cultures of Arnebia Euchroma Employing Two-Liquid-Phase Systems

1 INTRODUCTIONThe production of secondary metabolites using plant cells has been the subject of much inter-est in recent years.Despite that tremendous research efforts had been made in this topic,notmany products have reached the commercial stage.It is generally acknowledged that the mainproblem in this field is the lack of basic knowledge of the biosynthetic routes,and the mechanisms found to bring about the production of such secondary metabolites.There are,however,some techniques that have beneficial effects on the production and ex-

Clinical experimental study of Arnebia Root oil in increasing FGF expression and promoting wound surface healing

Objective：To investigate the effection of Arnebia Root oil on the FGF expression in wound surface and the ability to promote wound surface healing. Methods：24 wound surfaces of patients were divided into two groups. Experimental group was treated by Arnebia Root oil and the control was treated by petrolatum gauze. Histology, histochemistry, electron microscope methods and healing rate measurement were used to show the FGF expression and wound healing process. Results：Endogenous FGF were expressed in both of the groups, in which of the experimental group was higher than that of the control group, the wound surface healing rate of experimental group was also higher and paralleled with FGF expression. Conclusion：Arnebia Root oil has effects to promote FGF expression and enhances wound surface repair. The wound healing mechanism between the action of Arnebia Root oil and function of FGF need further investigating.

Effective assignment of positional isomers in dimeric shikonin and its analogs by ^(1)H NMR spectroscopy

An approach for distinguishing two types of positional isomers of dimeric shikonin and its analogs was explored with ^(4)JC,H long-range correlation by prolonging the acquisition time at ^(2,3)JC,H values of 2.0 and 8.0Hz.Furthermore,the ^(1)H(proton)nuclear magnetic resonance(NMR)pattern of phenolic hydroxyl protons was developed as a“diagnosis signal”to ascertain the relative location of each side chain in DMSO-d_(6) at sample concentrations of 0.022-0.034 mol/L.The chemical shift differences of 0.6ppm between OH-5' and OH-1 and between OH-8'and OH-4 are assigned to Type A and Type B,respectively.All reported ambiguous structures were corrected by this pattern.Additionally,the steric structures of isolated compounds were elucidated by quantum chemical calculations of electronic circular dichroism(ECD)spectra.

不同生境条件下软紫草(Arnebia euchroma)结实特性的差异

对软紫草在自然生境及人工栽培条件下的结实结籽特性进行比较分析。结果表明：软紫草不同居群、同一居群的不同生境以及同一生境的不同年份，在结实特性上均表现出不同程度的差异。自然居群软紫草植株的花数、结实率、结1～2粒和3粒种子的花的频率分布、种子产量、结籽率及种子千粒重均远低于人工栽培居群，彼此间存在极显著差异；自然居群各样地植株的花数、结实率、结1粒和2粒种子的花的频率分布、种子产量及结籽率均表现为样地S1〉样地S3〉样地S2，种子千粒重表现为样地S1〉样地S2〉样地S3，其中，结实率、种子产量、结籽率和种子千粒重在各样地间存在极显著差异。不同年份中，自然居群样地S1植株的结实、结籽率无显著差异，但花数、种子产量和种子千粒重存在极显著差异；而栽培居群植株的花数、结实结籽率、单株种子产量存在极显著差异，种子千粒重存在显著差异。

Arnequinol A and arnequinone A, two unique meroterpenoids from Arnebia euchroma

Arnequinol A(1),featuring an unprecedented 6/6/3 tricyclic carbon skeleton fused with a heptatomic oxo-bridge,together with arnequinone A(2)bearing a highly conjugated methyl-shifting benzogeijerene skeleton,were isolated from Arnebia euchroma.Their structures were elucidated by extensive spectro-scopic methods and quantum chemical calculations of the 13 C nuclear magnetic resonance(NMR)data and electronic circular dichroism(ECD)spectra.The plausible biosynthetic pathways for 1 and 2 were presented.In in vitro test,compound 2 showed potent neuroprotective activity against serum-deprivation induced PC12 cell damage at a concentration of 10μmol/L.