Background Proliferation of the human Tenon capsule fibroblasts(HTFs) is a main cause of failure of filtering surgery.To search the drug of inhibiting the growth of the HTFs is essential for the improvement of successful rate of filtering surgery.Objective The aim of this study was to investigate the inhibitory effect of apigenin on HTFs and its mechanism.Methods Human Tenon capsular tissue was obtained during the strabismus correction surgery.HTFs was primarily cultured using explant method and identified using vimentin by immunochemistry.The 3-5 generation of cells were incubated to 96-well plate.Apigenin of 0,20,40,80,160 μmol/L was added into the medium,respectively,for 24,48,72 hours,and the proliferation of HTFs was detected by sulfonyl chloride (SRB) at the wavelength of 560 nm (A560).Bromodeoxyuridine (BrdU) of 10 μg/L was added to culture the cells for 48 hours to calculate the labeling rate of BrdU.The morphology of the cells was observed using Hoechst 33258 staining,and apoptosis and cells cycle were evaluated by flow cytometry.Results Cultured cells grew well with the positive response for vimentin,showing the green fluorescence in cytoplasm.SRB assay showed that the A560 value was gradually declined with the increase of the dosage of apigenin and prolong of time (Fgroup =480.306,P =0.000 ; Ftime =555.144,P =0.000).The labeling rate after 0,40,80 μmol/L apigenin acted for 48 hours was (87.860 ±0.632)％,(61.520±4.306)％ and (23.480±4.472)％,showing a significant difference among the three groups (F =299.347,P =0.000).The labeling rate of HTFs for BrdU was significantly decreased in the 40 and 80 μmol/L apigenin groups compared with the 0 μmol/L apigenin group (P＜0.05).Hoechse 33258 staining found that the number of the HTFs was gradually decreased and the cell number of karyopyknosis and nuclear deformation was increased with the increase of apigenin dosage.Percentage of cells in G0/G1 phase were raised and that in S and G2/M phase were declined in the higher dosage apigenin group,with a significant difference among the different groups (FG0/G1 =58.621,P=0.000;Fs =32.357,P=0.001 ;FG2/M =83.998,P=0.000).In the 72nd hour after acted by 0,40,80,160 μmol/L apigenin,the apoptosis rate of HTFs was (4.77±0.21) ％,(13.24±1.35)％,(18.33±1.86) ％,(31.58 ± 2.77) ％,respectively,with a statistically significant difference among the four groups (F =204.791,P＜0.05).Conclusions Apigenin restrains the growth of HTFs by evoking G0/G1 cell cycle arrest and inducing apoptosis in a dosage-and time-dependent manner.

Prolyl-4-hydroxylase domain (PHDs) family is one of the most important regulatory factors in hypoxic stress. PHD2 plays a critical role in cells and tissues adaptation to the low oxygen environment. Its hydroxylation activity regulates the stability and transcriptional activity of the hypoxia-inducible factor 1 (HIF-1), which is the key factor in response to hypoxic stress. Subsequently, PHD2 acts as an important factor in oxygen homeostasis. Studies have shown that PHD2, through its regulation on HIF-1, plays an important role in the post-ischemic neovascularization. Furthermore, under hypoxic condition, PHD2 also regulates other pathways that positively regulate angiogenesis factors HIF-1 independently. Moreover, recently, several evidences have also shown that PHD2 also affects tumor growth and metastasis in a tumor microenvironment. Based on these facts, PHD2 have been considered as a potential therapeutic target both in treating ischemic diseases and tumors. Here, we review the molecular regulation mechanism of PHD2 and its physiological and pathological functions. We focus on the role of PHD2 in both therapeutic angiogenesis for ischemic disease and tumor angiogenesis, and the current progress in utilizing PHD2 as a therapeutic target.
Animals ; Humans ; Hydroxylation ; Hypoxia-Inducible Factor 1 ; metabolism ; Hypoxia-Inducible Factor-Proline Dioxygenases ; antagonists & inhibitors ; physiology ; Neoplasms ; blood supply ; metabolism ; pathology ; therapy ; Neovascularization, Pathologic ; metabolism ; pathology ; Tumor Microenvironment ; Vascular Diseases ; pathology ; therapy

0.05).Compared with the CSS,GMFM and WV before treatment,there were statistically difference after 6 and 12 weeks treatment in two groups(Pa

Aged ; Aged, 80 and over ; Anti-Bacterial Agents ; pharmacology ; Coal Mining ; Cross Infection ; complications ; microbiology ; Drug Therapy, Combination ; therapeutic use ; Humans ; Male ; Methicillin ; pharmacology ; Methicillin Resistance ; Microbial Sensitivity Tests ; Middle Aged ; Pneumonia ; drug therapy ; etiology ; Silicosis ; classification ; complications ; Staphylococcus aureus ; drug effects ; Treatment Outcome

Child ; Female ; Humans ; Menisci, Tibial ; abnormalities ; surgery

Objective To investigate the effect and mechanism of transplantation of marrow multi- function stem ceils in treating femoral head necrosis.Methods Sixty japanese rabbits were divided into A,B and C groups randomly.After creation of the models of hormone induced necrosis of femoral head;A group was designated as the treatment,B as the control and C as the normal groups.The bone marrow of A group was extracted and isolated and then injected into the left femoral head and the right femoral head was decompressed by drilling only.The rabbits were killed at 8 weeks after the treatment and changes in various parameters were observed,including imaging data of molybdenum target films,CT and MRI;routine pathology with HE staining and ultrastructural alteration by election microscopy.Results Eight weeks after the treatment of transplantation of marrow multifunction stem cells,the X-ray showed only a little change but the typical appearances were revealed by CT and MRI.Pathohistologic manifestation demonstrated decrease of empty bone lacuna,increase of osteoblast and new bone formation.Election microscopy displayed abundant organelles in osteoblasts with few empty bone lacuna,in addition the tansplantation of marrow multifunction stem cells could obtain better reconstraction for the involved femoral head.Conclusions The treatment of transplantation of marrow muhifunction stem cells in femoral head necrosis could accelerate the process of repairing,worthy to be acknowledged as a good and valuable management in rabbits.(J Intervent Radial,2007,16:122-126)

Objective: To observe the therapeutic efficacy of acupoint application at different groups of acupoints in treating bronchial asthma in remission stage. Methods:A total of 120 patients with bronchial asthma in remission stage were recruited and divided by the random number table method into acupoint application group 1, acupoint application group 2 and acupoint application group 3, with 40 cases in each group. In all the three groups, Tiantu (CV 22), Dazhui (GV 14) and Feishu (BL 13) were selected, with Dingchuan (EX-B 1) added in acupoint application group 1, Shenshu (BL 23) added in acupoint application group 2, and Gaohuang (BL 43) added in acupoint application group 3. Before intervention, one month and 3 months after intervention, clinical symptoms, peak expiratory flow (PEF) andforced expiratory volume in 1 second percentage of predicted value (FEV1％) of the three groups were observed, and their clinical efficacies were evaluated. Results: Comparing the therapeutic efficacy regarding traditional Chinese medicine symptoms and signs, after 1-month treatment, the total effective rate was 87.5％ in acupoint application group 1, versus 62.5％ in acupoint application group 2 and 55.0％ in acupoint application group 3, and the between-group differences were statistically significant. After 3-month treatment, the total effective rate was 95.0％ in acupoint application group 1, versus 70.0％ in acupoint application group 2 and 65.0％ in acupoint application group 3, and the between-group differences were statistically significant. After intervention, the three groups all showed significant improvements in pulmonary function with statistical significance; among the three groups, the improvement in acupoint application group 1 was more significant than that in the other two groups. Conclusion: Tiantu (CV 22), Dazhui (GV 14) and Feishu (BL 13) as basic prescription plus Dingchuan (EX-B 1) can improve symptoms of bronchial asthma in remission stage, and it works better in improving pulmonary function than the basic prescription plus Shenshu (BL 23) or Gaohuang (BL 43).

To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.
Animals ; Brain ; metabolism ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Iridoids ; chemistry ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

The key and crucial step of metabolic engineering during quinic acid biosynthesize using shikimic acid pathway is high expression of quinate 5-dehydrogenase. The gene qa-3 which code quinate 5-dehydrogenase from Neurospora crassa doesn't express in Escherichia coli. By contrast with codon usage in Escherichia coli, there are 27 rare codons in qa-3, including eight AGG/AGA (Arg) and nine GGG (Gly). Two AGG are joined together (called box R) and some GGG codons are relative concentrate (called box G). Along with the secondary structure of mRNA analysed in computer, the free energy of mRNA changes a lot from -374.3 kJ/mol to least -80.5 kJ/mol when some bases in the end of qa-3 were transformed, and moreover, the change of free energy is quite small when only some bases in the box G and box R transformed. After the change of rare codon and optimization of some bases in the end, qa-3 was expression in E. coli and also the enzyme activity of quinate 5-dehydrogenase can be surveyed accurately. All the work above benefit the further research on producing quinic acid engineering bacterium.
Alcohol Oxidoreductases ; biosynthesis ; genetics ; Base Sequence ; Codon ; chemistry ; genetics ; Escherichia coli ; genetics ; metabolism ; Hydro-Lyases ; genetics ; Molecular Sequence Data ; Neurospora crassa ; enzymology ; genetics ; RNA, Messenger ; chemistry ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Shikimic Acid ; metabolism ; Transformation, Bacterial

Children with autism spectrum disorder(ASD)may suffer from motor skill dysfunction,and perform as motor delay,coordination disorder,fitness decline,visual-motor integration disorder and so on,which disturb their daily life,school-based activities,and other social activities. Due to the obvious core symptoms of children with ASD,the problem of motor skills may be ignored. Children with ASD should be monitored for the motor development at their early stage and be evaluated with proper assessment tools. Children with ASD should be trained with individualized plan according to their features in order to develop motor skills and improve the core symptoms. As evidence of motor intervention for children with ASD is increasing,we should pay more attention to the motor skill dysfunction problem in children with ASD and make them benefit from the motor intervention and sport program.