The prescriptions containing pericarpium citri reticulatae that built by Professor. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. After analyzed by data mining, such as apriori algorithm, the frequency of single medicine, the frequency of drug combination, the association rules between drugs and core drug combinations can be get from the database. Through the analysis of 1 027 prescriptions with pericarpium citri reticulatae, these prescriptions were commonly used to treat stomach aches, cough and other syndromes. The most frequency drug combinations were "Citri Reticulatae Pericarpium-Poria", "Paeoniae Radix Rubra-Citri Reticulatae Pericarpium" and so on. The drug association rules that the confidence was 1 were "Glycyrrhizae Radix ex Rhizoma --> Citri Reticulatae Pericarpium", "Paeoniae Alba Radix-Cyperi Rhizoma --> Citri Reticulatae Pericarpium", "Poria --> Citri Reticulatae Pericarpium", and so on. The drugs in the prescriptions containing pericarpium citri reticulatae that built by Professor Yan mostly had the effects of regulating the flow of Qi and invigorate blood circulation, which reflected the clearly thought when making prescriptions.
Citrus ; chemistry ; Data Mining ; Drug Interactions ; Drug Prescriptions ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Practice Patterns, Physicians'

Objective To analyze our clinical results of head and neck reconstruction using microsur- gical free flap transfer techniques.Methods The free flap donor sites with long vascular pedicle and large diameter of vessel were routinely chosed,and chose receipt vessels with large diameter and proper position, and perform vessel ananstomosis under surgical loups instead of microscope.The un-buried free flap with a mo- nitoring window were harvest,and do double venous anastomoses in some flaps to ensure adequate venous out- flow.Results From May 1999 to March 2005,1066 consecutive free flap transfers were used to reconstruct head and neck defects.The overall success rate of free flap was 98.3%.The vessel thrombosis rate was 3.1%,and the flap salvage rate was 45.5%.Conclusion Head and neck reconstruetion using microsurgi- cal free flap transfer technique is safe and reliable,and good clinical results can be obtained.

Objective To investigate protein expression of telomerase reverse transcriptase(TERT),telomerase-associated protein 1(TP1) and telomeric repeat binding factor 2(TRF2) in human fetal cadiacmyocytes.Methods Using immunohistochemical method,sections of human fetal hearts were stained with antibodies against TERT,TP1 and TRF2.Results Expression of TERT was gradually decreased with development in human fetal cadiacmyocytes nuclear.Expression of TP1 and TRF2 was gradually increased with development in human fetal cadiacmyocytes cytoplasm.Conclusion Down-regulation of TERT and up-regulation of TP1 and TRF2 may promote the permanent withdrawal of cardiomyocytes from the cell cycle and enter terminal differentiation and myocardium hypertrophy.

· AIM :To investigate the role of pericytes in growth of retinal microvascular endothelial cells with a co-culture system in order to understand some mechanism of angiogenesis in hypoxia induced retinal neovascular disorders.(RMECs) were isolated by a modified protocol using CD31 coated Dynabeads, and identified by immunocytochemical staining with anti-Factor Ⅷ and CD31 antibodies. Rat retinal pericytes were isolated and characterized by immunofluorescent staining with PDGFR-β; and desmin antibodies. Pericytes and RMECs were cultured in a contact co-culture system both under normoxia and hypoxia by Millicell chamber. RMECs proliferation was evaluated by MTT and cell cycle assay with flow cytometry. RT-PCR was used to detect the alteration of KDR/Flk-1 mRNA level in RMECs under normoxia or hypoxia in the co-culture system.harvested with the modified isolating method. The two cell types were identified by positive Factor Ⅷ, CD31 and PDGFR-β, desmin cytochemical staining respectively.RMECs proliferated significantly under hypoxia from 3 to 9d with a maximal rate on day 6 (24.9%, P ＜ 0.01) by MTT. In the co-culture system, the proliferation of RMECs was inhibited by pericytes. After 6d exposure to hypoxia,the fraction of S-phase RMECs number was greatly increased by 43.9% (P ＜ 0.01). In the co-culture system,RMECs proliferation was inhibited by pericytes through decreasing the fraction of S-phase cell number both under normoxia (3.6%, P＜0.05) and under hypoxia (15.1%,P＜0.01). KDR/Flk-1 mRNA level in single cultured RMECs was shown to increase approximately 1.3-fold when exposed to hypoxia. Compared with single cultured RMECs, co-culture with pericytes could decrease KDR/Flk-1 mRNA by 45.1% (P＜0.05) and 27.7% (P ＜ 0.05) under normoxia and hypoxia condition respectively.pericytes could inhibit proliferation of RMECs under both normoxia and hypoxia. The inhibition effects of pericytes maybe, at least in part, due to downregulation of KDR/Flk-1 of RMECs. These findings confirm that pericytes could be a potential inhibitor in the pathogenesis of retinal neovascularization.

Humans ; Genetic Markers ; Family ; DNA Fingerprinting ; Microsatellite Repeats/genetics*

Demyelination is the pathological hallmark of multiple sclerosis (MS) lesions. The concept of remyelination has gained acceptance in recent years, but naturally occurring remyelination is incomplete. To improve repair processes, a number of strategies have been explored experimentally and clinical trials are being carried out. In principle, remyelination can be achieved by either promoting endogenous repair mechanisms or by providing an exogenous source of myelinating cells via transplantation. Both approaches have been successful in animal models of demyelination. In addition, many studies have elucidated the principal mechanisms of oligodendrocyte biology and remyelination in the central nervous system (CNS). Here, we review the neuroscientific background to the development of strategies for myelin repair and draw on a variety of more recent experimental findings to speculate on the likely evolution of remyelinating therapies in the future.

The prescriptions including Polygoni Multiflori Caulis that built by Pro. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. The method of association rules with apriori algorithm was used to achieve frequency of single medicine, frequency of drug combinations, association rules between drugs and core drug combinations. The datamining results indicated that in the prescriptions that including Polygoni Multiflori Caulis, the highest frequency used drugs were parched Ziziphi Spinosae Semen, Ostreae Concha, Ossis Mastodi Fossilia, Salviae Miltiorrhizae Radix Et Rhizoma, Paeoniae Rubra Radix, and so on. The most frequent drug combinations were "Polygoni Multiflori Caulis-parched Ziziphi Spinosae Semen", "Ostreae Concha-Polygoni Multiflori Caulis", and "Polygoni Multiflori Caulis-Ossis Mastodi Fossilia". The drug association rules of confidence coefficient 1 were "Ostreae Concha-->Polygoni Multiflori Caulis", "Poria-->Polygoni Multiflori Caulis", "parched Ziziphi Spinosae Semen-->Polygoni Multiflori Caulis", and "Paeoniae Alba Radix-->Polygoni Multiflori Caulis". The core drug combinations in the treatment of insomnia were Ossis Mastodi Fossilia, Polygoni Multiflori Caulis, Salviae Miltiorrhizae Radix et Rhizoma, Ostreae Concha, Polygalae Radix, Margaritifera Concha, Poria, and parched Ziziphi Spinosae Semen. And the core drug combinations in the treatment of obstruction of Qi in chest were Salviae Miltiorrhizae Radix Et Rhizoma, Polygoni Multiflori Caulis, parched Ziziphi Spinosae Semen, Trichosanthis Fructus, Allii Macrostemonis Bulbus, and Paeoniae Rubra Radix.
Data Mining ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional ; methods ; Plant Stems ; chemistry ; Polygonaceae ; chemistry ; Prescriptions

·AIM: To investigate the preparation of endostatin protein and its biologic activity on vascular endothelial cell.· METHODS: pBlast-hEndostatin and pBlast-Mcs were identified by digesting with Nhe Ⅰ and Sal Ⅰ, by PCR reaction, by sequencing, and by Alignments of PCR products with gene bank using NCBIBLAST software. The identified pBlast-hEndostatin as well as pBlast-Mcs were then purified with QIAGEN Endofree plasmid maxi kit.The purified plasmids transfected human fibroblasts. The expression of endostatin was detected by RT-PCR, Westem-Blot and immunohistochemistry. The endostatin prorein produced by transfected fibroblasts was purified by ultrafiltration and affinity chromatography. The inhibitory action of endostatin on human umbilical vein endothelium was measured by MTT assay.· RESULTS: pBlast-hEndostatin was found to contain human endostatin gene. Endostatin protein was produced by transfected fibroblasts. The inhibitory ratio of 2.5,5,10,20,40,80mg/L endostatin on human umbilical vein endothelium for 48h were 8.5%,13.1%,27.7%,38.1%,56.7%,63.8% respectively. IC50 value was 34.5mg/L.No inhibition action was found on fibroblasts.·CONCLUSIONS: Endostatin protein can be produced by the transfected fibroblasts. The produced endostatin has inhibitory action on human umbilical vein endothelium and has no inhibition action on fibroblasts.

OBJECTIVETo observe the effect of Fn-TPO gene modification on human bone marrow mesenchymal stem cells (MSCs).

METHODSRetroviral vector containing Fn-TPO gene was constructed and bone marrow MSCs was modified by this vector. The transcription of Fn-TPO gene in MSCs was observed. The proliferation capacities, hematopoietic cells adhering capacities and TPO secretion capacities of gene modified MSCs were assayed respectively. Cord blood CD34 cells were seeded on the gene modified MSCs layers and several essential growth factors were added. After co-culturing in vitro for 7 days, the number of CD34 cells and their colony forming capacities were assayed by flow cytometry and semisolid culture assay.

RESULTSRetroviral vector containing Fn-TPO gene was successfully constructed and bone marrow MSCs were modified by this vector. Fn-TPO gene was expressed by bone marrow MSCs after gene modification. The viability of MSCs had no significant difference between pre- and post-gene-modification [(7.18 +/- 0.89) 10(4)/ml vs. (6.92 +/- 0.77) 10(4)/ml, P > 0.05]. The hematopoietic cells adhering ability of gene modified bone marrow MSCs was reinforced(0. 188 +/- 0.018 vs. 0.167 +/- 0.017, P < 0.01). The concentration of TPO in the MSCs culture supernatant raised from (5.58 +/- 0.37) ng/ml to (7.46 +/- 0.59) ng/ml (P < 0.01) and did not significantly decline in a short-time period, but influenced by the growth status of MSCs. After co-culturing with gene modified MSCs for 7 days, the absolute number of nucleated cells, the percentage of CD34+ cells and the colony numbers of BFU-E, CFU-GM, CFU-GEMM were (29.9 +/- 2.7) x 10(4), (33.3 +/- 2.8)% , 109.3 +/- 4.1, 163.7 +/- 7.1, 13.3 +/- 1.5, respectively, being significantly higher than that co-cultured with non-modified MSCs.

CONCLUSIONSFn-TPO gene modification can improve the capacity of human bone marrow MSCs for hematopoietic cells adhering, TPO secretion and cord blood CD34 cells amplification.

Bone Marrow Cells ; cytology ; metabolism ; Cell Adhesion ; Cell Proliferation ; Cells, Cultured ; Fibronectins ; genetics ; Gene Fusion ; Genetic Vectors ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Retroviridae ; genetics ; Thrombopoietin ; genetics ; metabolism ; Transfection