Objective To establish a HPLC method for the determination of daphnetin, 7-OH coumarin and syringing in Zushima. Methods The separation was performed on YMC-Pack ODS (4.6 mm?250 mm, 5 ?m) column with a mobile phase consisting of methanol-0.5% acetic acid-water (19︰8︰1) as gradient elution at the flow rate of 1.0 mL/min. The dual detective wavelength was set at 327 nm and 266 nm. The column temperature was 30 ℃. Results Baseline separation was achieved for the analysis of the three index components. The average recoveries were 98.4%～100.2%. Conclusion This method is convenient, rapid and accurate. It can be used for quality control in production of Zushima.

Objective To evaluate the promotional effectiveness of different TCM appropriate technology packages; To provide a scientific basis for improving the technology package. Methods 8 assessment indexes, including input costs, the number of experts, the number of people with senior professional post, the number of grassroots people under the guidance of experts, the number of training hours, the number of people receiving training, evaluating rate and the number of users were selected. TOPSIS method was used to evaluate the promotional effectiveness of five kinds of TCM packages in grassroots. Results The Ci values for appropriate technology packages were 0.76 for pains in neck, shoulder, waist and lower extremities, 0.49 for gynecological diseases, 0.44 for infantile diarrhea, 0.66 for chronic gastropathy, and 0.00 for tumors. Conclusion The effectiveness of promotion of TCM appropriate technology packages from high to low is pains in neck, shoulder, waist and lower extremities package, chronic gastropathy package, gynecological diseases package, infantile diarrhea package, tumors package.

Objective To evaluate the clinical efficacy of Yangyin Yiqi mixture and western medicine in the treatment of oral lichen planus(OLP)and its effect on the levels of interferon -γ(IFN -γ)and interleukine -10 (IL -10)in serum and saliva.Methods Sixty patients with OLP were randomly divided into control group(n =30) and treatment group(n =30).The control group was treated with triamcinolone acetonide and lidocaine,local injec-tion.The treatment group was treated with Yangyin Yiqi mixture on the basis of the control group.The treatment lasted two months.After treatment,the clinical efficacy,objective indicators score,levels of IFN -γand IL -10 in serum and saliva,adverse reactions were compared between the two groups.Results The overall response rate of the treatment group (86.67%)was significantly higher than that of the control group (63.33%)(χ2 =11.64,P <0.01).After treatment,the two groups'objective index score significantly reduced(t =8.52,12.51,all P <0.01),and the treat-ment group's objective indicators rating was significantly lower than the control group (t =4.38,P <0.05).After treatment,the levels of IFN -γin serum and saliva were (25.39 ±1.29)pg/mL and (12.76 ±1.28)pg/mL,which were significantly increased in the treatment group (t =10.35,8.15,all P <0.01),so did the control group (P <0.05).And the levels of IL -10 in serum and saliva were (27.54 ±1.82)pg/mL and (9.92 ±0.86)pg/mL,and the levels were significantly decreased in the two groups after treatment (t =8.76,9.39,all P <0.01,t =4.65,4.94,all P <0.05).And levels of IFN -γin serum and saliva of the treatment group were significantly higher than those of the control group (t =4.68,4.32,all P <0.05),and levels of IL -10 in serum and saliva of the treatment group were significantly lower than those of the control group (t =5.41,5.25,all P <0.05).During treatment,there were no sig-nificant adverse reactions.Conclusion Yangyin Yiqi mixture combined with western medicine has definitive clinical efficacy in the treatment of OLP.It can significantly increase the level of IFN -γand reduce the level of IL -10 in serum and saliva in patients,so provide theory value for treatment of OLP.

Objective To investigate the effect of songling xuemaikang(SL-xmk)pretreatment on the expression of matrix metalloproteinase-9(MMP-9)after focal cerebral ischemia-reperfusion in rats.Methods A total of 45 male Sprague-Dawley rats were randomly allocated into SL-xmk pretreatment,sham operation,and normal saline control group.Preventive gavage was per-formed for 8 weeks in rats using SL-xmk(937.50 mg/kg)suspension in the SLxmk pretreatment group(n = 15);the preventive gavage was performed in rats using the equal volume of normal saline in the sham operation(n = 15)and normal saline control(n = 15)groups.At the end of the pretreatment process,a model of middle cerebral artery occlusion(MCAO)in rats was induced by suture method for 2 hours and reperfusion for 24 hours.The effects of SL-xmk pretreatment on the neurologic deficit scores after transient MCAO,brain water content,and infarct volume in rats were observed.Immunohistochemical staining was used to detect the MMP-9 immunoreactive positive cells in ischemic brain tissue.Results The neurologjc deficit scores(1.21 ± 0.25 vs.2.37 ± 0.35,P = 0.000),the brain water content (76.24% ± 7.09% vs.88.78% ± 6.57%,P = 0.000),the percentage of infarct volume (22.62% ±2.17% vs.27.84% ±3.43%,P =0.000),and the numbers of MMP-9 positive cells(16.20 ± 2.17/mm vs.20.60 ± 2.71/mm,P = 0.000)were all significantly lower than those in the control group.Conclusions SL-xmk pretreatment may significantly inhibit the expression of MMP-9 in the brain tissue of focal cerebral ischemia-reperfusion rats and reduce brain water content and infarct volume.

Objective To investigate the feature brain damage and clinical manifestations in neuromyelitis optica (NMO) patients; To investigate the relationship between serum NMO-IgG antibody and NMO brain damage. Methods Clinical data of 37 NMO patients and their head and spinal cord MRI by 1.5T superconducting MR scanner, were analyzed; serum NMO-IgG antibody were measured by immunofluorescence. Results 17 cases were found to have abnormal signals on MRI, which were mainly in the white matter, pons, medulla, ventricle, aqueduct, and around the corpus callosum; According to pathological changes, brain damage can be divided into scattered irregularity (13 cases), fusion (3 cases),multiple sclerosis-like (1 case) ,with scattered irregularity more common,5 cases had clinical manifestations of brain damage: somnolence, vomiting, diplopia, visual rotation, 11 cases patients with brainstem damage show positive serum NMO-IgG antibodies. Conclusions Brain damage can be seen in half of NMO patients, they often located in the high expression area of AQP4: brain white matter, periventricular,brainstem and so on. Clinical symptoms has nothing to do with the size of lesions but the location, they often occur when brainstem was involved. Serum NMO-IgG is helpful in differentiating NMO with brain damage and MS.

Objective:To optimize the culture system for the mesenchymal stem cells from the Placenta Chorion (hCMSC) in vitro by adding the selected cytokines to amplify the hCMSC effectively,with furthermore some preliminary analysis of the hCMSC performed.Methods:Immunohistochemistry was applied to show the localization of the mesenchymal stem cells within the placenta chorion,then the corresponding part of chorion was digested by collagenase Ⅳ to get the cell suspension for primary culture and subculture;Flow Cytometry was used to detect the surface marker of hCMSC,whose proliferation was detected by direct cell counting and the MTT method;then Butylated Hydroxyanisole (BHA),and DMSO was used combinedly as the inducer in the differentiation of the hCMSC to the neuron like cells,which was then detected by Immunofluorescence staining for the expression of NSE;finally RT-PCR was used to detect the expression of flt3.Results:The result of immunohistochemistry showed CD105~-,CD90~- positive hCMSC was much easier to be found around the blood vessels within the villous of chorion,which then could be obtained by mechanical dissociation and collagenase digestion.bFGF was good at amplifying the hCMSC among all selected cytokines,but FL was showed even obviously effective on the proliferation of the hCMSC.The propagating cells were found to highly express CD73,CD90,CD105;and negative for CD14,CD34,CD45 and HLA-DR.We also found that the hCMSC happened to express PDL-1 the negative co-stimulatory molecules,while their potential to different into the neuron like cells was still well maintained.Conclusion::There is low immunogenicity mesenchymal stem cells within the placenta chorion,both bFGF,FL are shown good effects on the proliferation of the hCMSC,with maintaining their differentiation potential as well.

Objective To detect changes of regional grey matter and white matter volume in patients of neuromyelitis optica (NMO) by voxel-based morphometry (VBM),and investigate its relationship with clinical variables.Methods Conventional magnetic resonance imaging (MRI) and structural threedimensional MRI were obtained from 20 NMO and 20 sex-and age-matched healthy volunteers.The comparison of grey matter and white matter volume between the two groups was analyzed by VBM tools of statistical parametric mapping (SPM) 5.Pearson correlational analysis was used to assess correlations between regional volume decrease and disease duration and expanded disability status scale (EDSS) scores in NMO patients.Results Compared with normal controls,NMO patients had grey matter atrophy in several cortical regions,such as right inferior frontal gyrus (cluster size 514),left superior temporal gyrus (282),right middle temporal gyvus (229) and right insula (211) (t =3.58-5.11,AlphaSim corrected,P ＜0.05).White matter atrophy was found in several subcortical regions in NMO patients,such as right precentral and postcentral gyrus (cluster size 457,110),left middle frontal gyrus (285),and right inferior parietal lobule (231) (t =2.90-4.25,AlphaSim corrected,P ＜ 0.05).Grey matter and white matter volume loss were not significantly correlated with clinical duration or EDSS score in NMO.Conclusion By means of VBM,regional atrophy of grey matter and white matter is found in NMO patients,which may provide evidence for brain structural abnormality in NMO.

Objective To investigate the feature of regional grey matter volume changes in relapsing-remitting multiple sclerosis (RRMS) patients by voxel-based morphometry ( VBM) and presume the possible pathophysiological basis.Methods Conventional magnetic resonance imaging (MRI) and T1-weighted three-dimensional MRI were obtained from 32 RRMS and 32 sex- and age-matched normal controls.The comparison of grey matter volume between the two groups was analyzed by statistical analysis software SPM5 and VBM.A Pearson correlational analysis was used to assess correlation between gre matter loss and disease duration,expanded disability status scale (EDSS) and visible brain lesion volume.Results Compared with normal controls,RRMS patients had extensive bilateral grey matter atrophy in thalami (left 2031 and right 1711),caudate (left 815 and right 1031) and parahippocampal gyrus (left 313 and right 467),as well as several cortical regions in frontal,temporal,parietal,and occipital lobes (t value were between 8.853 and 11.163,all P ＜ 0.01).Regional grey matter loss in bilateral thalami ( r value were - 0.596 on left and were - 0.694 on right) and right caudate ( r = - 0.409 ) were strongly negatively correlated with visible brain lesion volume in RRMS (all P ＜ 0.05 ).Conclusions By means of VBM,extensive grey matter atrophy are found in RRMS patients,especially in deep grey matter.Axonal degeneration secondary to visible brain lesions may be a key pathogenesis of grey matter atrophy in RRMS.

OBJECTIVEUtilizing a gene reporter technique to study the effects of Andrographitis Herba on human CXCR4 and CCR5 promoters.

METHODInhibition of CXCR4 and CCR5 on T cells of healthy volunteers was analyzed by RT PCR, Western blot and flow cytometry. The human CXCR4 and CCR5 promoters driving a luciferase reporter in vectors pGLA. 17-CXCR4 and pGLA. 17-CCR5 were transfected into H9 stem cells. G418 was used for selecting stable cell lines. Rat sera thus medicated was collected and added to the transfected H9 cells, in which the expression of CXCR4 and CCR5 promoters was detected.

RESULTThey showed that the mRNA and protein expression levels of CXCR4 and CCR5 in human CD4+ T cells decreased significantly after taking Andrographitis Herba (P<0.05). Furthermore human CXCR4 and CCR5 promoter activity was downregulated significantly by sera from rats medicated with Andrographitis Herba.

CONCLUSIONAndrographitis Herba may have the effect of down-regulating CXCR4 and CCR5 promoters. It provides a feasible experimental platform for screening herbal medicine as the treatment of HIV/AIDS.

Adult ; Andrographis ; chemistry ; Animals ; CD4-Positive T-Lymphocytes ; drug effects ; metabolism ; Cell Line ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Promoter Regions, Genetic ; drug effects ; Rats ; Rats, Wistar ; Receptors, CCR5 ; genetics ; metabolism ; Receptors, CXCR4 ; genetics ; metabolism ; Young Adult

Objective:To develop an intervention program for symptom clusters in children with acute lymphoblastic leukemia (ALL) during the consolidation and reinforcement phase based on symptom management theory from a sociopsychological perspective.Methods:An initial draft of the intervention program for symptom clusters in children with ALL during the consolidation and reinforcement phase was prepared through literature review and group discussions. Purposeful sampling was used to select 20 experts as respondents for a Delphi method consultation. After two rounds of expert consultations, adjustments and improvements were made to the relevant indicators, resulting in the final version of the intervention program.Results:The effective response rates for the two rounds of expert consultations were 80% (16/20) and 100% (20/20), respectively. The authority coefficients for the experts were 0.816 and 0.838, while the Kendall's coordination coefficients were 0.207 and 0.397 ( P<0.01 for both rounds). The final intervention program comprised 3 primary items, 7 secondary items, and 27 tertiary items. Conclusions:The intervention program for symptom clusters in children with ALL during the consolidation and reinforcement phase, developed from a sociopsychological perspective, demonstrates scientific validity, reliability, and feasibility. It can offer guidance and reference for symptom management in children with ALL.