0.05). CONCLUSION: The degree of tolerance and interference on their sleep experienced by both the self-ligating brackets Smartclip and the conventional straight wire brackets Victroy groups are similar.

Objective To evaluate in vitro effects of propranolol and isoproterenol on proliferation of infantile hemangioma endothelial cells(IHECs)as well as expressions of vascular endothelial growth factors(VEGF and VEGF-2) and human basic fibroblast growth factor (bFGF). Methods The second - third passage endothelial cells derived from the proliferative phase of infantile hemangioma were divided into propranolol and isoproterenol groups. The propranolol group was further classified into 5 groups to be treated with propranolol solutions at concentrations of 10, 15, 20 mg/L, EGM-2 medium (blank control group 1), or EGM-2 medium containing 0.16% DMSO (DMSO group), while the isoproterenol group was classified into 4 groups to be treated with isoproterenol solutions at concentrations of 5, 10, 20 mg/L or EGM-2 medium(blank control group 2). Methyl thiazolyl tetrazolium(MTT)assay was performed to evaluate cellular proliferative activity in these propranolol groups at 24, 48, 72 and 96 hours separately, enzyme-linked immunosorbent assay (ELISA)to measure VEGF, VEGF-2 and bFGF lev els in culture supernatants of IHECs at 24 and 48 hours separately. Results The proliferative activity of IHECs showed no significant differences between 10-, 15- and 20-mg/L propranolol groups at either 24 or 48 hours (H = 1.152, 2.643, respectively, both P > 0.05), or between the blank control group 1, DMSO group, and 10- and 15-mg/L propranolol groups at either 72 or 96 hours, but significantly decreased in the 20-mg/L propranolol group compared with the blank control group 1 at 72 and 96 hours (both P < 0.05). The 24-hour treatments with propranolol or isoproterenol at the above concentrations all affected the expressions of VEGF, VEGF-2 and bFGF to different degrees. At 48 hours, there was a significant decrease in VEGF levels in 15- and 20-mg/L propranolol groups, as well as in VEGF-2 and bFGF levels in 10-, 15- and 20-mg/L propranolol groups compared with the blank control group 1 (all P < 0.05), but a significant increase in VEGF levels in 5-, 10- and 20-mg/L isoproterenol groups compared with the blank control group 2 (all P < 0.05), as well as in VEGF-2 and bFGF levels in the 20-mg/L isoproterenol group compared with the blank control group 2, 5- and 10-mg/L isoproterenol groups (all P < 0.05). Conclusions The treatment with propranolol at certain concentrations for certain durations can suppress the growth of, as well as expressions of VEGF, VEGF-2 and bFGF in, endothelial cells derived from the proliferative phase of infantile hemangioma, whereas that with isoproterenol has opposite effects. The therapeutic mechanism of propranolol in infantile hemangioma may be associated with expressions of β-adrenergic receptors and their downstream signal transduction-related cytokines.

Objective To explore the role of beta-2 adrenergic receptor in the pathogenesis of infantile hemangioma.Methods In vitro cultured infantile hemangioma endothelial cells were divided into propranolol and isoproterenol groups.The propranolol groups were further classified into 5 groups to be treated with propranolol solutions at concentrations of 10,15,20 μg/ml,EGM-2 medium (blank control group 1),and EGM-2 medium containing 0.16％ DMSO (DMSO group) respectively,while the isoproterenol groups were classified into 4 groups to be treated with isoproterenol solutions at concentrations of 5,10,20 μg/ml and EGM-2 medium (blank control group 2) respectively.After 24-and 48-hour treatment,enzyme-linked immunosorbent assay (ELISA) was performed to measure the expression of beta-2 adrenergic receptor in cell culture supernatants in the above groups.Results After 24-hour treatment,15-μg/ml and 20-μg/ml propranolol groups showed significantly decreased expression of beta-2 adrenergic receptor compared with blank control group 1 and DMSO group (all P ＜ 0.05).After 48-hour treatment,all the propranolol groups showed significantly decreased expression of beta-2 adrenergic receptor compared with the blank control group 1 (all P ＜ 0.05).However,the expression of beta-2 adrenergic receptor was significantly higher in the 10-and 20-μg/ml isoproterenol groups than in the blank control group 2 after 24-hour treatment (all P ＜ 0.05),and higher in the 20-μg/ml isoproterenol group than in the blank control group 2 after 48-hour treatment (P ＜ 0.05).Conclusion Propranolol can down-regulate the expression of beta-2 adrenergic receptor on the surface of vascular endothelial cells,while isoproterenol can up-regulate its expression.

Objective:To evaluate the effect of laser irradiation on related growth factors in and apoptosis of in vitro cultured infantile hemangioma endothelial cells. Methods:Cultured infantile hemangioma endothelial cells were divided into 3 groups: intense pulsed light (IPL) group irradiated with IPL at a dose of 23 J/cm 2 for 1 session, laser group irradiated with 1 064-nm Nd:YAG laser at a dose of 90 J/cm 2 for 1 session, control group receiving no irradiation. On days 1, 3 and 7 after irradiation, RT-PCR was performed to measure the mRNA expression of vascular endothelial growth factor (VEGF) , VEGF receptor 2 (VEGFR-2) and basic fibroblast growth factor (bFGF) , and Western blot analysis to determine VEGFR-2 protein expression, enzyme-linked immunosorbent assay to detect the levels of VEGF and bFGF in the culture supernatant, and flow cytometry to detect cell apoptosis. Results:Compared with the control group, the laser group showed significantly decreased mRNA expression of VEGF (0.363±0.021 vs. 1.000±0.023, P< 0.001) , VEGFR-2 (0.483±0.017 vs. 1.001±0.031, P=0.001) and bFGF (0.402±0.040 vs. 1.000±0.004, P< 0.001) , decreased VEGFR-2 protein expression (0.332±0.055 vs. 0.768±0.096, P< 0.05) , decreased levels of VEGF (69.389±24.179 ng/L vs. 334.506±13.084 ng/L, P< 0.001) and bFGF (2.386±0.151 ng/L vs. 9.165±0.232 ng/L, P< 0.001) in the culture supernatant, but significantly increased apoptosis rate (18.413%±2.654% vs. 4.300%±0.036%, P< 0.01) on day 7 after irradiation. Compared with the control group, the IPL group also showed significantly decreased mRNA expression of VEGF (0.436±0.041 vs. 1.000±0.023, P< 0.05) , VEGFR-2 (0.493±0.037 vs. 1.001±0.031, P< 0.05) and bFGF (0.490±0.044 vs. 1.000±0.004, P< 0.05) , decreased VEGFR-2 protein expression (0.406±0.037 vs. 0.768±0.096, P< 0.05) , decreased levels of VEGF (128.858±6.063 ng/L vs. 334.506±13.084 ng/L, P< 0.001) and bFGF (2.723±0.471 ng/L vs. 9.165±0.232 ng/L, P< 0.001) in the culture supernatant, but significantly increased apoptosis rate (16.597%±1.877% vs. 4.300%±0.036%, P< 0.01) on day 7 after irradiation. Conclusion:The 1 064-nm Nd:YAG laser may exert a therapeutic effect on hemangioma by inhibiting hemangioma endothelial cells via regulating key factors on the VEGF/VEGFR-2 signaling pathway and cell apoptosis.

Objective　To study the correlation between the rabbit glucocorticoid receptor(GR) and steroid-induced glaucoma(SIG).Methods　The GR binding in peripheral blood leukocytes and plasma cortisol were measured in 15 adults pigmented rabbits on the first day of experiment, 0.5mg dexamethasone was injected in the right eyes of each rabbit once every 2 days, totally 15 times for 30 days. Right eyes intraocular pressure(IOP) and aqueous outflow facility were measured every week, the relation between GR, plasma cortisol, IOP and aqueous outflow facility(C) was analyzed. Results　After the experiment the IOP increased by 0.67kPa±0.49kPa(P＜0.01),C decreased by 0.08±0.11(P＜0.01).GR may closely be related to IOP and C(r=0.56,P＜0.025; r=0.60,P＜0.01).Plasma cortisol was independent of IOP and C(r=0.27, P＞0.05;r=0.34,P＞0.05).Conclusion　The level of GR has positive correlation with the sensitivity of tissue of eye to glucocorticoid.

ObjectiveTo study the therapeutic effect of variable pulsed Nd∶YAG laser and intense pulsed light (IPL) on port wine stain(PWS) in an animal model.MethodsThe combs of 38 Leghorn chickens served as the animal model of PWS,which were randomly classified into 4 groups:blank control group receiving no irradiation,Nd:YAG group receiving Nd∶YAG laser irradiation for 2 sessions,IPL group receiving IPL irradiation for 2 sessions,and combination group receiving 1 session of Nd∶YAG laser irradiation followed by 1 session of IPL irradiation 45 days later.One week after the final irradiation,the rnorphology of combs was observed,and tissue specimens were resected from the combs for the analysis of histological changes and blood vessel density.The data were assessed by using one-way ANOVA.ResultsAfter 2 sessions of irradiation,the color of comb skin was lightened,and light microscopy showed the thickening of vascular layers in the dermis,blockage of blood vessels and decrease in the number of blood vessels.The count of blood vessels per high power field(× 400) was significantly lower in the Nd∶YAG group(17.92 ± 3.63),IPL group(8.08 ± 1.56)and combination group(7.08 ± 1.31) than in the control group(37.08 ± 3.97,all P ＜ 0.01 ),but was similar between the combination group and IPL group (P ＞ 0.01),and higher in the Nd∶YAG group than in the combination group(P ＜ 0.01 ).ConclusionsVariable pulsed Nd∶YAG laser and IPL,alone or in combination,can be used in the treatment of PWS,however,the effect of IPL alone or in combination with Nd:YAG seems superior to that of Nd∶YAG laser alone.

Objective:To establish the determination method for ATP, ADP and AMP in the myocardial tissue of rats and research the changes of ATP, ADP and AMP in heart fallure rats. Methods: A Thermo Hypersil C18 (250 mm × 4. 6 mm,5 μm) column was used. The solution of 0. 68% potassium dihydrogen phosphate and 0. 116% soduim hydroxide was applied as the mobilie phase; The detection wavelength was at 254 nm. The SD rats randomly divided into 2 groups received saline and adriamycin by intraperitoneal in-jection for 6 weeks, respectively. The heart tissue was withdrawn for homogenate. The contents of ATP, ADP and AMP in rat heart tis-sue were determined by HPLC. Results:The calibration curves were linear from 0. 625μg·ml-1 to 40μg·ml-1 for ATP, ADP and AMP. The limit of detection and limit of quantification were 0. 625μg·ml-1(r>0. 99). The precision of RSD was less than 9. 50%. The accuracy was 93. 2%-108. 0%. The method stability (RSD) was from 3. 0% to 14. 0%. The content of ATP in heart fallure mod-el group induced by adriamycin was significantly lower than that in saline group,while that of ADP were much higher than that in saline group(P<0. 01) and AMP. Conclusion:The HPLC method for the content determination of ATP, ADP and AMP in myocardial tissue meets the requirements of the determination of biological samples, which can be used to study the effects of anthracycline-based antitu-mor drugs in heart fallure rats.

PURPOSE To investigate the application of CLSM in tumor pathology with the three dimensional reconstruction by confocal laser scanning microscopy in routine pathologic specimens of HCC, METHODS 30 ?m thick sections were cut from the paraffin-embedded tissues of HCC. hyperplasia and normal liver, stained with the DNA fluorescent probe YOYO-1 iodide and examined by CLSM to collect optical sections and 3D reconstructed images. RESULTS HCC displayed chaotic arrangement of carcinoma cell nuclei, marked pleomorphism. indented and irregular nuclear surface, and irregular and coarse chromatin texture. CONCLUSION The serial optical tomograms of CLSM can be used to create 3D reconstruction of cancer cells. Such 3D impressions might be helpful or even essential in arriving at an exact diagnosis

Objective To investigate the expression and the early diagnostic significance of heat shock protein 70 in acute allograft rejection of liver-transplamed rats. Methods The model of rat orthotopic liver transplantation was made by using a modified "two-cuff technique". The rats were randomly divided into 3 groups. For each group, donors and receptors all included 15 rats respectively. The control group: Wistar to Wistar liver transplantation; The untreated group: SD to Wistar liver transplantation, not receiving any immunosuppressant after liver transplantation; The treatment group: SD to Wistar liver transplantation, receiving intramuscular injection of tacrolimus (FKS06, 2mg kg-1. day-1) after operation. Five rats were executed randomly in every group on the post-transplantation day 3, 5 and 7 and the graft samples were obtained for optical microscopic observation. The expression of HSPT0 in grafts was detected by using immunohistochemical method and RT-PCR. The correlation between acute rejection following liver transplantation and the expression of HSP70 in grafted liver was studied. Results There was no acute rejection examined in the control group. The untreated group showed typical allograft rejection and the rejection activity index (RIA) went up gradually after the operation (P<0.01). The treatment group showed no rejection or borderline allograft rejection. The level of HSP70 was increased transiently after operation, then reduced in the control group (P<0.05). The level of HSP70 in the untreated group was higher than in the control groupand gradually increased with the prolongation of time after transplantation (P<0.01). A significant correlation was found between HSP70 and pathological score in the untreated group (P<0.01). The treatment group showed low levels of HSP70 of all the time. Conclusions The expression of HSP70 in grafts is closely related to the occurrence and development of the acute rejection and can be useful for early diagnosis of acute allograft rejection following liver transplantation.

Objective:To analyze the drug use of a patient with intestinal obstruction induced by high dose of morphine to explore the role of clinical pharmacists during the therapeutic process. Methods:The dose titration of morphine, choice of analgesic drugs and dose conversion were adjusted by doctors and pharmacists using NRS scores of dynamic assessment for the patient. The pharmaceutical care was implemented for the patient induced by high dose of morphine. Results:The pharmaceutical supervision by clinical pharma-cists improved medication compliance of the patient. After fentanyl patches were used to replace morphine, the intestinal obstruction was relieved in the patient and the quality of life was improved. Conclusion:Clinical pharmacist can promote the rational use of opi-oids for patients with intestinal obstruction. The successful treatment of the patient with intestinal obstruction provides the reference for patients with intractable pain treated by opioids.