Objective To investigate the effect of acteoside on injury PC12 cells induced by glutamate. Methods PC12 cells were assigned into normal control group, model control group, positive drug group and acteoside(AS) treated group. Every group was treated by 1. 5 mmol·L-1 glutamate for 24 hours except for the control group, and the injury was antagonized by 10 μmol·L-1 Vit E and acteoside at different concentration(15. 625, 31. 25, 62. 5, 125 and 250 μmol·L-1 ). Cell morphology was observed by inverted microscope, cell survival was determined with MTT, LDH activity was measured by enzyme label kit, the MDA content and SOD activity were measured by TBA kit and WST kit, and the ROS was measured by Elisa kit. Results Compared with the model control group, all doses of acteoside could significantly improve the PC12 cell morphology and survival (P<0. 05), inhibit LDH activity and production of MDA and ROS (P<0. 05), increase the activity of SOD (P<0. 05), except for the lowest dose group. Conclusion Acteoside has protective effects on PC12 cells injured by glutamate.

Objective To investigate the effect of Galangin of different purity on melanocyte proliferation and melanin synthesis of A375-HaCaT co-culture system. Methods The A375-HaCaT co-culture system was established with Transwell technology, and 0.1, 0.5, 1.0, 5.0, 10 μg/mL Galangin of the purity of 70%and 90%was used to act on the co-culture system. Cell proliferation, melanin content and tyrosinase of A375 were measured by MTT assay, NaOH lysis assay and L-DOPA oxidation assay respectively. The cytokines such as ET-1 and SCF in HaCaT were detected by ELISA. Results A375 cells in co-culture system grew well, and 0.1, 0.5, 1.0, 5.0, 10 μg/mL Galangin of the purity of 70% and 90% had up-regulation effect on cell proliferation, melanin content and tyrosinase of A375. Moreover, Galangin could increase the expression level of ET-1 and SCF of HaCaT at more than 0.5 μg/mL, and the effect of Galangin of 70% purity was better than 90% purity. Conclusion Galangin could promote the cell proliferation, melanin content and tyrosinase of A375, and mechanism of the pathways is probably related to the up-regulation on the expression of ET-1 and SCF of HaCaT.

Objective To optimize the best condition of semi-bionic extraction for Vernonia anthelmintica.Methods The best condition of the semi-bionic extraction for V.anthelmintica by uniform design was optimized with total HPLC integrate area of common peaks in fingerprints,total flavonoids and dried extract weight as the indexes.Results According to industry production condition,the best technical condition:pH values of 80% ethanol for the 1st,2nd,and 3rd decoctions were in order of 6.0,6.5,and 8.0,and the extraction time were 1,0.5,and 0.5 h,respectively.Conclusion It is better for V.anthelmintica extracted by semi-bionic extraction,and the parametrs are optimized by uniform design.

Aim To investigate the effect of acteoside (AS)on the expression of caspase-3 in cerebral cortex of mouse models of Alzheimer’s disease(AD).Meth-ods Kunming (KM)strain mice were assigned into control group,model group,positive control group (VitE)and acteoside group.Every group was induced by a combination of D-galactose(i.p.60mg·kg -1 · d -1 )and AlCl3 (i.g.5mg·kg -1 ·d -1 )for 60ds ex-cept for control group,then mice were treated by dif-ferent concentrations(30,60,1 20 mg·kg -1 ·d -1 )of acteoside for 30ds.During the time,mice were in-duced continuously by a combination of D-galactose and AlCl3 .The learning and memory of mice were de-tected by step-down test,the activity of AChE in serum and brain of mice was measured by chemical colorime-try,the structure changes in cerebral cortex were ob-served by HE staining,and the expression of caspase-3 in cerebral cortex was analyzed through the immunohis-tochemical staining.Results Compared with model group,acteoside could improve the learning and mem-ory abilities(P <0.05 or P <0.01 ),decrease the ac-tivity of AChE in serum and brain(P <0.05 or P <0.01 ),and improve the morphology and number of neuron in cerebral cortex(P <0.01 ).Moreover,acte-oside could significantly inhibit the expression of caspase-3 in cerebral cortex (P <0.05,P <0.01 ). Conclusion Acteoside has significantly protective effects on brain damage of mice induced by a combina-tion of D-galactose and AlCl3 , and it′s protective mechanism probably relate to inhibiting the expression of caspase-3 and maintainings the normal morphology and number of neuron in cerebral cortex.

OBJECTIVE:To investigate the in vitro transdermal absorption properties of galangin and effects of different pene-tration enhancers on its transdermal behaviors,and provide reference for developing skin preparations using galangin as APIs in the treatment of vitiligo. METHODS:HPLC was used to determine the galangin content. Using cumulative permeation rate (Q) and the transdermal rate(J)of galangin as indexes,the effect of absorption of receiving solution [20%,40% polyethylene glycol 400 (PEG400)solution and 30% ethanol solution] and rotating rate(200,300,400 r/min)on galangin in complete skin of mice were investigated,as well as the azone(1%,3%,5%)and propylene glycol(10%,20%,40%)alone or combination on its penetra-tion promotion. And the transdermal properties of galangin in complete skin,exfoliating skin,dermis skin of rats and mice were de-tected. RESULTS:The best permeability of complete skin of mice showed in 40% PEG400 solution at rotating speed of 300 r/min with 5% azone alone,J was 3.2570 μg/(cm2·h). Js of complete skin,exfoliating skin,dermis skin of mice were 2.7199,34.016, 33.874 μg/(cm2·h),respectively;and those of rats were 0.4996,9.5124,17.406 μg/(cm2·h). CONCLUSIONS:Galangin can penetrate the complete skin of mice and rats,however,the penetration quantity is far lower than exfoliating skin and dermis skin.

ObjectiveTo investigate the effects of extracts ofLiaoxuan Kaxifu Powders (LE) on proliferation and transcription of IL-8 and ICAM-1 in HaCaT induced by TNF-α; To discuss its mechanism of action.Methods Cultured HaCaT was assigned into normal group, TNF-α group and low-, medium- and high-dose of LE group. Every group was induced by 40 ng/mL TNF-α except for normal group, and then LE groups were treated by different concentrations (8, 40, 200μg/mL) of LE for 48 h. The proliferation of HaCaT was evaluated by MTT and the apoptosis was detected by inverted fluorescence microscope. Levels of IL-8 and ICAM-1 in HaCaT were assessed by ELISA, and their mRNA expressions was detected by semi-quantitative RT-PCR.Results Compared with normal group, the absorbency of HaCaT and the contents and mRNA expressions of IL-8 and ICAM-1 increased in TNF-α group (P<0.05,P<0.01); compared with TNF-α group, LE of all dose groups could significantly inhibit the absorbency, decrease the contents and mRNA expressions of IL-8 and ICAM-1 (P<0.05,P<0.01).Conclusion LE is able to inhibit the proliferation of HaCaT induced by TNF-α, and the mechanism is probably related to promoting apoptosis and down-regulating the gene expressions of IL-8 and ICAM-1, and then maintaining the normal level of HaCaT.

Objective To observe the features of a rare Miao pedigree of Bardet-Biedl syndrome in Yunnan province and to reserve its gene. Methods Three patients of this pedigree were reported. Their clinical and biochemical features were compared with those of the other pedigree members. Lymphocytes from main members of this pedigree were collected and transformed with cyclosporine A methods. Immortalized B lymphocyte strains were checked by means of chromosome karyotype analysis. Results Patients of this pedigree demonstrated typical clinical characteristics of this syndrome with increased body weight, blood pressure, fasting glucose, and lipoprotein(a)as compared with the other pedigree members(P＜0. 05). The chromosome karyotype of the lymphocytes before and after transformation was kept consistent. Conclusions Patients of this Miao pedigree showed typical clinical characteristics of this syndrome as well as abnormal metabolic features. Immortalized B lymphocyte strains with their genetic information were set successfully.

Objective To analyze the clinical manifestations of primary Sj(o)gren's syndrome (pSS)with peripheral neuropathies.Methods Eighty-six patients who fulfilled the 2002 American-European Consensus Group criteria for pSS were enrolled in the study.For each patient,medical data,including clinical,laboratory,immunologic and electromyography data were collected and analyzed.The clinical manifestations of primary Sj(o)gren's syndrome were compared between patients with and without peripheral neuropathy.Statistical methods used were t-test,chi-square test and Logistic regression.Results Eighty-six patients were analyzed,and neurological involvement was noted in 26％ (22/86) patients.The clinical spectrum of peripheral neuropathies encountered in Sj(o)gren's syndrome patients was wide,with sensory neuropathies being the most common.Median nerve,peroneal nerve and sural nerve were the most likely involved,and lower limb involvement accounted for 73％ (16/22).Peripheral neuropathy was diagnosed during the Sj(o)gren's syndrome course in all patients,and about 45％ patients' neurological involvement were diagnosed early in the course of the disease.The frequency of Raynaud's phenomenon was significantly higher (32％ vs 5％,P=0.002) as well as acroanesthesia (68％ vs 5％,P＜0.01) in pSS with peripheral neurological involvement than in pSS without peripheral neuropathy.The median values of EULAR Sj(o)gren's syndrome disease activity index (ESSDAI) were 5.3 (range 2.8-7.8) and 3.4 (range 1.5-5.3) in the PNS and non-PNS groups respectively (P＜0.01).We found a significant rise of peripheral neuropathy risk associated with Raynaud's phenomenon (relative risk 9.489,95％CI 2.191-41.093,P=0.003) and ESSDAI (relative risk 1.528,95％CI 1.179-1.979,P=0.001).Elevated titers of rheumatoid factor (P=0.023) and ANA (P=0.003) were common in patients with peripheral neuropathy.Conclusion Peripheral neuropathy is not a rare manifestation of pSS.Neurological involvement can be diagnosed early in the course of the disease.Raynaud's phenomenon and high disease activity may be the risk factors for peripheral neuropathy.

Objective To evaluate the expression of RAR-? gene in cervical carcinoma cell lines SiHa,HeLa,C33A and Caski and to analyze the relation between their gene expression and the promoter methylation of RAR-? DNA.Methods The expression of mRNA and protein of RAR-? gene in the four cell lines were analyzed by RT-PCR,western blot and immunofluoreseence,respectively.Methylation specific PCR(MSP)was used to check whether there was methylation in the promoter of RAR-? gene.The demethylating agent 5-aza-2'-deoxycytidine(5-Aza-cdR)was used to treat methylated cell lines and the change of RAR-? gene methylation and RAR-? gene expression defects were observed.The cell proliferation was assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.Results The mRNA and protein expression levels of RAR-? in cell lines SiHa,HeLa,Caski and C33A were 0.25 ?0.08,0,0.60?0.19,3.12?0.92 and 0.23?0.07,0,0.14?0.05,0.68?0.21,respectively.The mRNA and protein expression of RAR-? in SiHa,HeLa and Caski cell lines were decreased or silenced, whereas its expression increased in C33A cell line.MSP method showed that there were RAR-? gene methylation in SiHa,HeLa and Caski cell lines,while there was no RAR-? gene methylation in C33A cell line.After treated with 5-Aza-cdR,the mRNA and protein expression levels of RAR-? in SiHa,HeLa, Caski and C33A cell lines were 1.82?0.59,2.13?0.62,1.67?0.43,2.95?0.89 and 0.69?0.21, 0.83?0.29,0.56?0.16,0.64?0.20 respectively.The mRNA and protein levels of RAR-? had a significant difference between before and after interference with 5-Aza-cdR in SiHa,Helm,and Caski cell lines(P0.05).The 5-Aza-cdR treatment could suppress cell proliferation.Conclusions The RAR-? gene expression defects play an important role in the carcinogenesis of cervical cancer.Aberrant methylation in promotor region of RAR-? gene may be an important mechanism for the loss of expression of RAR-? gene.

AIM: To analyze the genotype of the allele distribution of a polymorphic G-954C within the 5 upstream promoter region of the nitric oxide synthetase 2A gene (NOS2A) in samples of diabetic retinopathy in patients with cystoid macular edema in the mainland of China.METHODS: Eighty-nine patients with diabetic retinopathy and cystoid macular edema and 90 healthy controls were enrolled in this study. Nest polymerase chain reaction (PCR)was performed, and restriction endonudease digestion and gene fragments sequence were examined to detect the genotype of NOS24 G-954C.RESULTS: The genotypes of the sample population of 89 cases and 90 healthy controls were all detected as GG.CONCLUSION: The distribution of G-954C of NOS2A polymorphism are at a lower frequency in China, with little relevancy to the frequency of diabetic retinopathy combined with cystoid macular edema.