Objective To analyze the genetic relationship among the different cultivars of Cornus officinalis and provide some foundation for heredity breeding. Methods A modified method of extracting total DNA from the leaves of C. officinalis was selected by improving the traditional method-CTAB, the total DNA was analyzed by RAPD; the genetic similarity correlation was calculated by SPSS 10.0 DICE method, cluster analyses were carried out using Between-groups linkage method, and the genetic dendrogram was established. Results Random primers (22 10-bp) were selected to be used for the PCR, a total of 133 bands were amplified by 12 samples, among which 75 bands were polymorphic, accounting for 56.39%. The cluster analysis indicated that: spindleform itself was one group, short pear-shape and short cylindericform clustered together, the rest longer types clustered together, which reflected the result of artificial selection in the course of cultivating. Conclusion The result keeps accordance with the biologic character and territorial distribution of C. officinalis cultivars; RAPD analysis is an assistant mean for seed breeding of C. officinalis.

This paper was aimed to explore the relationship between different commercial specification and chemical constituents on Radix Rehmanni,which provided experimental evidences for drawing up the commodity commercial specification and grading criteria of Radix Rehmanni.Several classified indexes,including grade,surface color,section color,section texture were carried to explore the change rules of chemical constituents by setting the contents of iridoid glycoside,sugars,water content,total ashes,acid-insoluble ashes and water extract as the markers.The results showed that the grade,surface color and section texture had less impact on chemical constituents.However,the section color had more influence.Along with the section color gradually being deepened,the contents of iridoid glycoside and oligosaccharides of Radix Rehmanni were lower,and the contents of monosaccharide and polysaccharide were higher.It was concluded that there was correlation between commercial specification and chemical constituents of Radix Rehmanni.

SAbstact:Objective To establish a method for detecting t he pharmacokinetic parameters of te tramethylpyragine(TMPz ),the main active component in Xiongbing nasal spray(A),Chuanxiong nasal spray(B)and Xiongbing decoction(C),in the rat brain .Methods Gas chromatography was applied for pharmacokinetic analysis of TMPz in v arious time after nasal administration or gastr ic infusion.Results The concentration -time process of A can be described in an open two -department model,which is simi lar to the process of intravenous adm inistration.As compared with B and C,A has its characteristics:quick absorpt ion,quick distribution and quick excretion of TMPz.TMPz passed through blood -brain barrier almost right after administration,and reach the peak concentra tion 1minute after the administrati on.95.7%of TMPz was cleared from brain within 1.5h.Conclusion The application of Borneolum makes T MPz of A enter into the rebral tissue easily.Therefore Xiongbing nasal spray is a quick -effective pre paration for acute ischemic stoke an d can be medicated repeatedly.

OBJECTIVE:To establish a method for identification of Cornus officinalis and its Preparations. METHODS: LC-MS method was adopted. The separation was performed on Hypersil C18 column (150 mm?2.1 mm,5 ?m) with a mobile phase of methanol-acetic acid(84 ∶ 16) at a flow rate of 0.2 mL?min-1. The column temperature was set at 30℃. Mass spectra were equipped with an electrospray ionization (ESI) source with negative ion detection model as ion scanning. The conditions of determination were sheath gas flow rate of 1.5 L?min-1,sweep gas flow rate of 0.45 L?min-1,spraying voltage of 4.5 kV, high temperature capillary temperature of 350℃ and scan range of 110～600 amu. RESULTS: The three-level mass spectrometry of the ursolic acid and oleanolic acid was well separated. The results were stable. Three-level mass spectrometry of ursolic acid and oleanolic acid were established and identification of C. officinalis preparations was carried out.CONCLUSION:Three-level fragment peaks of fat-soluble constituents of C. officinalis can be used for indentification of C. officinalis and its preparations.

Objective To investigate the correlation between expression of four adhesion molecules in sera and in skin lesions of patients with psoriasis vulgaris, as well as their relevance to clinical severity of the disease. Methods Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum levels of ICAM-1, ICAM-3, VCAM-1 and ELAM-1 in 36 patients with plaque type psoriasis vulgaris and 36 healthy controls. Avidin biotin immunoperoxidase staining system was employed to quantitate expression of the four adhesion molecules in psoriatic skin lesions before and after treatment. Results Significantly increased expression of four adhesion molecules was found in psoriatic skin lesions (P

OBJECTIVETo establish sequence characterized amplified region markers of Cornus officinalis and provide a scientific basis for molecular identification of C. officinalis.

METHODThe random primer was screened through RAPD to obtain specific RAPD marker bands. The RAPD marker bands were separated, extracted, cloned and sequenced. Both ends of the sequence of RAPD marker bands were determined. A pair of specific primers was designed for conventional PCR reaction, and SCAR marker was acquired.

RESULTFour pairs of primers were designed based on the sequence of RAPD marker bands. The DNA of the seven varieties of C. officinalis was amplified by using YST38 and YST43 primer. The results showed that seven varieties of C. officinalis were able to produce a single PCR product. It was an effective way to identify C. officinalis. The varieties with cylindrical and long-pear shape fruits amplified by YST38 showed a specific band, which could be used as the evidence of variety identification. Seven varieties of C. oficinalis were amplified by using primer YST39. But the size of band of the variety with spindly shape fruit (35,0400 bp) was about 300 bp, which was shorter than those of the variety with the other shape fruits of C. officinalis (650-700 bp). The variety with the spindly shape fruit could be identified through this difference. The primer YST92 could produce a fragment from 600-700 bp in the varieties with cylindrical and long-pear shape fruits, a fragment from 200-300 bp in the varieties with oval and short-cylindrical shape fruits and had no fragment in the varieties with long cylindrical, elliptic and short-pear shape fruits, which could be used to select the different shapes of C. officinalis.

CONCLUSIONSCAR mark is established and can be used as the basis for breeding and distinguishing the verieties of C. officinalis.

To separate and identify the chemical constituents from the leaves of Broussonetia papyrifera (Linn.) Vent, various columns including Diaion HP-20, Toyopearl HW-40C, Sephadex LH-20, silica gel were employed for the isolation and purification of compounds from the leaves of B.papyrifera. The structures of the compounds were elucidated by their physiochemical characteristics and spectral data. Nineteen compounds were isolated from the leaves of B.papyrifera and their structures were identified as apigenin (1), apigenin-7-O-β-D-glucopyranoside (2), chrysoerid-7-O-β-D-glucopyranoside (3), apigenin-7-O-β-D-glucopyranuronide (4), vitexin-7-O-β-D-glucopyranoside (5), luteolin (6), 5,7,4′-trihydroxyl-6-C-[a-L-rhamnopyranosyl(1→2)]-β-D-glucopyranosyl flavone (7), 5,7,4′-trihydroxyl-8-C-[a-L-rhamnopyranosyl(1→2)]-β-D-glucopyranosyl flavone (8), saponaretin (9), vitexin (10), benzyl benzoate-2,6-di-O-β-D-glucopyranoside (11), (2R,3R,5R,6S,9R)-3-hydroxy-5,6-epoxy-β-ionol-2-O-β-D-glucopyranoside (12), (2R,3R,5R,6S,9R)-3-hydroxyl-5,6-epoxy-acetyl-β-ionol-2-O-β-D-glucopyranoside (13), ficustriol (14), (6S,9S)-roseoside (15), 3β-hydroxy-5α,6α-epoxy-β-ionone-2α-O-β-D-glucopyranoside (16), icariside B1 (17), sammangaoside A (18), 3-hydroxy-5α,6α-epoxy-β-ionone (19). Compounds 11, 12 and 13 are new compounds, the others are isolated from this genus Broussonetia for the first time.

Objective To measure the expression of activin receptor-like kinases 1(ALK1)in dermal fibroblasts from patients with systemic scleroderma(SSc)and to estimate its role in the production of fibronectin and plasminogen activator inhibitor-1(PAI-1).Methods Dermal fibroblasts were isolated from the lesions of 12 patients with SSc as well as the normal skin of 14 healthy controls,and subjected to a primary culture.The third-passage fibroblasts were used in the next experiment.Western blot and indirect immunofluorescence technique were utilized to quantify the expression of ALK1.A specific siRNA targeting ALK1 was designed,constructed,and transiently transfected into the control dermal fibroblasts,which were then classified into 2 groups to be cultured with or without the presence of transforming growth factor(TGF)-β1 for 72 hours followed by the detection of fibronectin and PAI-1 expression with Western blot.Results As Western blot and direct immunofluorescence technique showed,both control and SSc fibroblasts showed an expression of ALK1 in the cytoplasm and membrane,and the expression intensity of ALK1 in SSc fibroblasts was significantly higher than that in the control fibroblasts(1.97 ± 0.05 vs.1.12 ± 0.03,t =50.96,P ＜ 0.05).The expression of ALK1,fibronectin and PAI-1 was decreased by 90％,58％ and 31％ respectively in specific siRNA-transfected SSc fibroblasts compared with the control siRNA-transfected fibroblasts.TGFβ1 significantly increased the expression of ALK1,fibronectin and PAI-1 in the control siRNA-transfected fibroblasts,but the increase was markedly inhibited by the siRNA-targeting ALK1.Conlusion TGFβ1 can promote the production of fibronectin and PAI-1 via ALK1 in fibroblasts,and ALK1 may be involved in the development of sclerosis in SSc.

Carcinoma, Merkel Cell ; pathology ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Skin Neoplasms ; pathology

ObjectiveTo study the composition of volatile oil in Opisthopappus taihangensis （Taihangju）， and provide a reference for comprehensive development of this medicine. MethodTaking Chrysanthemum morifolium （Xiaobaiju）， C. morifolium （Xiaohuangju） and C. indicum （Yejuhua） as control， the qualitative and quantitative analysis of volatile oil in Taihangju and three control varieties were completed by gas chromatography-mass spectrometry （GC-MS） combined with retention index method. The GC conditions were as following：programmed temperature （initial temperature at 60 ℃， kept for 2 min； up to 120 ℃ with the heating rate of 5 ℃·min^-1， still kept for 2 min； up to 180 ℃ with the heating rate of 2 ℃·min^-1， kept for 3 min； and then up to 240 ℃ with the heating rate of 8 ℃·min^-1， kept for 5 min； finally up to 280 ℃ with the heating rate of 10 ℃·min^-1， kept it for 5 min and finished）， high-purity helium as the carrier gas， the split ratio of 50∶1. MS conditions were as follows：electron impact ion source （EI）， ion source temperature of 230 ℃， electron collision energy of 70 eV and scanning range of m/z 30-445. Principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to obtain the characteristic components between Taihangju and the three control varieties. ResultA total of 86， 96， 112 and 109 compounds including 73 common components were identified in Taihangju， Xiaobaiju， Xiaohuangju and Yejuhua， respectively. The contents of volatile components in Taihangju were significantly different from that of the control varieties. In which， the relative contents of α-thujone， eucalyptol and terpinen-4-ol were high in Taihangju， and eucalyptol， camphor and α-terpinyl acetate were the main compositions in the control varieties. In addition， 11 compounds were screened as characteristic components to distinguish Taihangju and the three control varieties by PCA and OPLS-DA， including main differential components of chamazulene and δ-cadinene. ConclusionThe main components of volatile oil in Taihangju includes alcohols， terpenes， ketones and esters with high medicinal value. The accuracy of qualitative analysis of volatile oil is improved by GC-MS combined with retention index method， which provides scientific reference for development and utilization of Taihangju.