Objective To investigate the changes of T cells and dendritic cells (DCs) and their related factors in children with immune thrombocytopenia (ITP) before and after therapy,and to analyze their clinical significance.Methods T-cells and DCs subsets were determined by flow cytometry both in 64 children with ITP (ITP group) before and after therapy and the control group.The serum levels of interleukin (IL)-4,interferon-γ (IFN-γ),transforming growth factor beta 1 (TGF-β1),and IL-27 were detected by enzyme linked immunosorbent assay (ELISA).Results Treatments of glucocorticoid or IVIg were effective in 41 cases of 64 ITP children.Compared to the control group,helper T cells (Th),Th/suppressor T cells (Ts),T regulatory cells (Treg),plasmacytoid DC (pDC),pDC/myeloid DC (mDC),and TGF-β1 in ITP patient group before treatment were significantly lower,while IFN-γ and IFN-γ/IL-4 were significantly higher (P ＜0.05).In ITP group,Th,Th/Ts,Treg,pDC,pDC/mDC,TGF-β1,and IL-27 were significantly increased,while IFN-γ and IFN-γ/IL-4 were decreased in children with ITP after therapy and achieved response (P ＜ 0.05).However,there was no significant difference between before and after therapy in ITP children without treatment response (P ＞ 0.05).Conclusions T cells and DCs subsets disorder and abnormal cytokine levels are observed in children with ITP,which can be corrected by immunosuppressive therapy,indicating that Th1 overactivity and the decrease of Treg and pDC both in quantity and function may be related to the pathogenesis of ITP in children.

BACKGROUND:Studies concerning bone marrow mesenchymal stem cells (BMSCs) cultured by umbilical cord serum in vitro were hot topic to avoid the heterogenous serum rejection during BMSC culture and improve culture efficiency of BMSCs.OBJECTIVE:To compare biological feature of BMSCs by the umbilical cord serum and adult autoserum in vitro.DESIGN,TIME AND SETTING:The opening experiment was performed at the Laboratory of Cell Biology,Xiangya Second Hospital,Central South University,China from October 2006 to June 2008.MATERIALS:Sixteen bone marrow samples were provided by Xiangya Second Hospital,Central South University and Xiangtan Central Hospital of Hunan Province.METHODS:BMSCs were obtained from 16 fresh adult bone marrow by gradient centrifugation with Ficoll Paque,cultured with DMEM/F12 with 10% umbilical cord blood serum and 10% adult autoserum.The fourth passage was used in this study.The surface antigens of BMSCs were detected by flow cytometry.BMSCs could differentiate into ostenblasts and adipocytes under culture in conditioned medium for osteogeneais and adipogenesis and the differentiated BMSCs were identified by morphological observation,immunophenotype and immunochemical staining.MAIN OUTCOME MEASURES:The following parameters were measured:morphological observation;cell cycle and cell count;identification of surface antigen;observation of osteoblasts and adipocytea induced from BMSCs by immunochemical staining.RESULTS:The quantity and velocity of BMSCs in umbilical curd blood serum was better than those in adult autoserum (P ＜ 0.05)Only few cells were proliferating in both medium,BMSCs in S phase in umbilical cord blood serum was more than those in adult autoserum (P ＜ 0.05).The positive rate of CD29,CD73 and CD105 on BMSCs in umbilical cord serum (above 90%) was higher than those in adult autoserum (P ＜ 0.05),and the positive rates of CD31,CD34 and CD45 were lower than 2%,and the positive rate of CD31 was lower than those in adult autoserum (P ＜ 0.05).The positive rate of BMSCs differentiated into osteoblasts and adipocytes in umbilical cord blood serum was also higher than these in adult autoserum (P ＜ 0.05).CONCLUSION:Purity and differentiated potency of BMSCs in umbilical cord blood serum are better than those in the adult autoserum in vitro.The umbilical cord serum is more adapt to clinical application.

Objective To investigate the surgical treatment and clinical outcome of patients with intracranial posterior circulation poor-grade aneurysms. Methods The clinical data of 35 patients with intracranial posterior circulation poor-grade aneurysms were collected. Seventeen patients were World Federation of Neurosurgical Societies (WFNS) grade IV and 18 patients were WFNS grade V. Twenty- nine patients were definitively treated with open microsurgery treatment(2 patients) and endovascular treatment (27 patients). Six patients received conservative treatment, including 4 patients only receiving external ventricular drainage to decrease intracranial pressure and 2 patients only receiving medical treatment. The nerve function were evaluated by Glasgow outcome scale (GOS) at discharge from hospital and 1 and 12 months after operation. Results In the patients who received conservative treatment, 5 patients were dead in hospital, and 1 patient got good outcome after 12 months. In the patients who received surgical treatment, at discharge from hospital good outcome were achieved in 15 patients (51.7%), poor outcome in 9 patients (31.0%), and death happened in 5 patients (17.2%). At 1 month after operation, good outcomes were achieved in 14 patients (48.3%), poor outcome in 9 patients (31.0%), and death happened in 6 patients (20.7%). At 12 months after operation, good outcome were achieved in 18 patients (62.1%), poor outcome in 5 patients (17.2%), and death happened in 6 patients (20.7%). Conclusions Patients with posterior circulation poor-grade aneurysms have better prognosis after positive surgical treatment.

Purpose:To study the effects and overexpression of p21 WAF1/CIP1 gene (p21 gene) on the SGC 7901 cell line plus cisplatin. Methods:An adenovirus expression vector with full length cDNA of p21 gene insert was constructed (ad p21) and transferred into SGC 7901 cell line with p53 mutation, the effects of ad p21 on the growth of SGC 7901 cell line was examined in vitro and in vivo . Results:The expression of p21 gene in SGC 7901 cell was confirmed by FCM, The growth of the Ad p21 gene transfected SGC 7901 cells was significantly inhibited as compared to mock (Ad LacZ) transfected SGC 7901 in vitro and induced the transfected cells appearing apoptotic, Combined with chemotherapeutic agent cisplatin (DDP), it could more strongly inhibit SGC 7901 cell growths.Conclusions:p21 gene can inhibit SGC 7901 cells growth and shows enhanced therapeutic efficiency for SGC 7901 cells when combined with DDP, especially in vivo experiment. Its use for gene therapy of cancer, especially when combined with other apoptosis inducing agents is promising.

AIM: To evaluate the efficacy of combined therapy for amblyopia in children by making use of pattern visual evoked potential ( P-VEP) game.
METHODS: This was a prospective case control study. These asthenopic children were divided into two groups. The control group ( 66 eyes of 49 patients ): occlusive therapy with glasses, cover, precision work, red light treatment and so on, later the stereo vision training was added. The experimental group (72 eyes of 52 patients):conventional methods mentioned above with P - VEP games.
RESULTS: The total effective rate and cure rate of experimental group in 6mo were higher than those of control group. The overall effective rate was 94. 4% in the experimental group and 83. 3% in the control group. There was a statistically significant difference between them (P<0. 05).
CONCLUSION: The comprehensive therapy by making use of P-VEP game is an individualized effective new way in treating amblyopia.

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 evaluate the effects of adeno-associated virus-enhanced green fluorescent protein (AAV-EGFP)on the biologic behavior of rabbit's bone marrow stromal cells(BMSCs)by means of a simple method of culturing and osteogenic induction in vitro so as to find an ideal viral vector and cell tracing mark for tissue en- gineering.Methods Total bone marrow culture was conducted to obtain rabbit BMSCs which were then induced in the osteogenic direction.The morphology of the cells was observed continuously,and their surface antigen and ossification were detected by alkali phosphatase stain and Von Kossa stain.On the basis of the above results, AAV-EGFP was transfected into the induced cells.The morphologic changes of the cells,the expression time and intensity of fluorescent light were observed.The transfection efficiency was detected to find the best multiplicity of infection(MOI)value.The cell growth curves were drawn to evaluate the biologic effects of AAV-EGFP on the cyto-activity.Results The morphology and purity of the rabbit BMSCs obtained were good.The ossification of the cells was significant after osteogenic induction.The best MOI value was found to be 1?10~5.The expression intensity of fluorescent light was strong with the expression time more than eight weeks so that the fluorescent light could be observed after cell generations.The transfection efficiency of AAV was high without significant biologic effects on the cyto-activity.Conclusions The total bone marrow culture and in vitro cell induction can satisfy the requirements for seeding cells in tissue engineering.AAV is an ideal viral vector for tissue engineering.Transfection of AAV-EGFP to cells could be an ideal method for cell tracing mark.

Objective To assess the application of three-dimensional digital subtraction angiography (3D-DSA) in evaluation of ruptured intracranial aneurysm after clipping and to discuss the different variable use of vol-ume rendering(VR), gradient rendering (GR) and maximum intensity projection (MIP). Methods From January 2011 to December 2012 , 88 patients with 92 ruptured intracranial aneurysms were treated with clipping using titani-um clips in our hospital and followed up by both 2D-DSA and 3D-DSA. Residual aneurysms , Clips place, clips and parent arteries and stenosis of parent arteries were evaluated by volume rendering (VR), gradient rendering (GR) and maximum intensity projection (MIP). Results Among 92 clipped aneurysms, 23 residual aneurysms were found by 3D-DSA. Residual aneurysms were recorded according to the Sindou grade: 15 of gradeⅠ, 3 of gradeⅡ, 4 of grade Ⅲand 1 of grade Ⅳ. Three patients of grade Ⅲand 1 of grade Ⅳwith residual aneurysms were retreated by clipping or coiling, and 1 patient of grade Ⅲ was dead with rupture of residual aneurysm. The clips and number of clips were clearly visualized , and relationship between the clips and the aneurysms was well demonstrated by VR, GR and MIP images. VR, GR images showed the remnants clearly. Three-dimensional digital subtraction angiography did not showed accurate details of the stenosis of parent arties which required an analysis of 2D-DSA. Conclusion Three-dimensional digital subtraction angiography can be used for definite evaluation of resid-ual aneurysms after clipping, especially by VR, GR images. It is helpful to manage the residual ruptured aneurysms.

This study was aimed to clone the GGPS (geranylgeranyl pyrophosphate synthase) gene from Lepidium apetalum, to analyze its sequence, and to express the protein in E.coli expression system. Specific PCR cloning primers were designed for GGPS gene from Lepidium apetalum according to the full-length sequence from a previous transcriptome sequencing project. PCR amplification was performed with this primer pair on a leaf cDNA template. TA cloning, sequencing and sequence analysis were performed.GGPS gene from Lepidium apetalum was expressed in the E.coli expression system. The results showed that the full-lengthGGPS cDNA from Lepidium apetalum was 1 146 bp coding a protein of 381 amino acids. The LaGGPS protein had an isoprenoid synthase domain. According to a phylogenetic tree constructed with multiple alignment of GGPS protein sequences from various plant species, GGPS protein from Lepidium apetalum was the closest to Arabidopsis thaliana and Sinapis alba. The prokaryotic expression vectorpET-32a-LaGGPS was also constructed successfully. The protein was expressed in E.coli BL21 strain. It was concluded that the cloning and prokaryotic expression of LaGGPS gene provided a foundation for a follow-up research of its function with protein purification and activity analysis.

AIM To investigate the effects of frost-like powder,steaming,stir-frying with wine,stir-frying with salt-water and stir-frying with vinegar on compositions and contents of fatty oils in Descurainiae Semen.METHODS Descurainiae Semen was processed by five methods,respectively.The fatty oils were extracted from various processed products by petroleum ether,which were then derivatized.GC-MS was adopted in the qualitative identification and quantitative determination.RESULTS Except for frost-like powder,various processing methods could increase the extraction rate of fatty oils.Compared with raw product,the quantities of fatty oils in various processed products were decreased,together with the increased contents.The main compositions of obtained fatty oils were unsaturated fatty acids,whose contents in various processed products (except stir-frying with vinegar product) were higher than those in the raw product.CONCLUSION The effects of different processing methods on compositions and contents of fatty oils in Descurainiae Semen show obvious differences,among which the processing effect of stir-frying with vinegar is not satisfactory.