Objective To observe clinical effect of alprostadil on DB artery of lower extremity scleratheroma block system patients.Methods To retrospectively analyze clinical data of lower extremity scleratheroma block system patients in our hospital,which were divided into detection group and control group.Results The bore 、blood flow and Vdias of femoral artery、popliteal artery、arteria tibialis posterior and dorsal artery of foot of detection group were better than control group,the clinical total effective rate of detection group was higher than control group (P ＜ 0.05 ),the difference was statistically significant.Conclusion The clinical curative effect of DB artery of lower extremity scleratheroma block system by alprostadil was obviously,which was worthy to be used.

Objective To explore the effect of cryotherapy and compression system in rehabilitation treatment for reconstruction of anterior cruciate ligament.Methods One hundred and twenty patients who had been treated with reconstruction of anterior cruciate ligament were randomly divided into treatment group (60 patients,treated with cryotherapy and compression system) and control group (60 patients,treated with traditional method).All patients was treated for 3 months and followed up for 6 months.The scores of visual analogue scales(VAS),degree of swelling,Lysholm and IKDC scores were observed and compared.Results The scores of VAS in treatment group postoperative 6,24 and 48 h were significantly lower than those in control group (P ＜ 0.01).The degree of swelling in two groups postoperative 6 h,12 h,24 h,48 h,1 week,2 weeks had significant difference(P ＜ 0.05).The degree of swelling in treatment group postoperative 4 weeks,3 months and 6 months was lower than that in control group,but there was no significant difference (P ＞0.05).The scores of Lysholm and IKDC in treatment group postoperative 4 weeks were (65.2 ± 12.3),(65.3 ± 14.3) scores,in control group were (60.2 ± 12.5),(59.4 ± 13.5) scores,there was significant difference (P ＜0.05).The scores of Lysholm and IKDC in two groups postoperative 3,6 months hod no significant difference (P ＞ 0.05).Conclusion The cryotherapy and compression system in rehabilitation treatment for reconstruction of anterior cruciate ligament can improve the effect of early-term rehabilitation training,but has little impact on the long-term rehabilitation training.

Objective To establish a cell model that is close to the HCV replication in vivo and can support long-term HCV replication in vitro.Methods A human liver cell line 7721 was inoculated with serum from a chronic hepatitis C patient for 8 hours.After incubation,the presence of HCV RNA,the expression of HCV antigen and the location of HCV RNA in cells and/or supernatant were detected by RTPCR,immunohistochemistry and in situ hybridization respectively.Results It was found that plus-and minus-strand of HCV RNA could be discontinuously detected in both cells and supernatant as late as 66 days after inoculation even if cells had been subcultured for 6 times.HCV NS3、NS5 antigens could be expressed in cells and HCV RNA was mainly located within cytoplasm.Conclusion The above results suggested that 7721 cell line was not only susceptible to HCV but also could support its long-term replication in vitro.This HCV infection model in vitro was proved to be a useful tool for studying the mechanism of HCV infection and replication,as well as evaluating the antiviral compounds and screening the protective antibodies/vaccines.

ObjectiveTo explore a procedure to correct the multiple facial deformities by using the expanded frontal flap combined with the axial flaps.MethodsAccording to the face deformity we used the rectangle 100-350 ml expanders behind the frontal hairline,after finishing the tissue expanding,adopted two or three axial flaps based on the supraoribital,supratrochlear or temporal vessels. ResultsA total of 13 cases were treated with this approach.7 eases were nose and lip defect reconstruction after burn,in which temporal vessel-based flap was used in 2 cases and supraoribital or supratrochlear vessel-based flaps in 5 cases.The other 6 cases were nasal reconstruction combined with the frontal defect correction by using random flaps,including 2 cases of pigmented nevus,1 neurofibroma,and 3 burn scars.All the flaps survived and satisfactory appearance was obtained.Conclusions The expanded frontal flap combined with axial flap based on multiple vessels is a good approach to correct the multiple facial deformities.

Objective:To evaluate the efficacy of occlusal rehabilitation in old patients with decreased vertical dimensions caused by serious attrition.Methods:Sixty-five patients with decreased vertical dimension caused by serious attrition were divided into 3 groups according to the defect of dentition and/or TMD.Group A included 42 patients with serious attrition and defection of dentition.Group B had 15 patients with heavy tooth wear,defection of dentition and TMD.Group C included 8 patients with serious attrition and functional TMD.All patients were conducted occlusal rehabilitation at increased vertical dimensions in normal intercuspital position(ICP) by removable denture with occlusal pad and removable occlusal splints.The time of follow-up visit of these patients ranged from 1.5 months to 9 years. Results:The masticatory efficacy of the patients was obviously improved,and all the symptoms of TMD were relieved.Furthermore,the symptoms of TMD were not observed after the treatment in patients without TMD.Conclusion:The results achieved by removable occlusal rehabilitation to aged people with serious attrition and decreased vertical dimension are satisfactory,and suggest that the occlusal rehabilitation is necessary for them.Furthermore,the removable occlusal rehabilitation can be acted as a reference and transition to permanent fixed occlusal rehabilitation.

Objective To screen and identify proteins interacting with hematopoietic stem/progenitor cell differentiation-related gene HSPC016, and to explore the molecular mechanism involved in the regulation by HSPC016 on the aggregative behavior of dermal papilla cells. Methods By using yeast two-hybridization,HSPC016 gene was sub-cloned into pGBKT7 to construct the bait plasmid (named as pGBKT7-HSPC016) in yeast AH109. The cDNA yeast expression library of human dermal papillae cells in yeast Y187 was screened with the bait plasmid and the proteins interacting with HSPC016 were identified. Yeast two-hybridization retransformation experiment was conducted to exclude the false positive clones and verify the interactions, then,the positive clones were sequenced and analyzed by using bioinformatic methods. Results The bait plasmid pGBKT7-HSPC016 was constructed successfully and there was no self-activation in or toxicity against yeast AH 109. Four proteins,including forkhead family of transcription factors (FOXO 1 ), mitogen-activated protein kinase 11 (MAPK 11 ), phosphoinositide-3-kinase (PIK3R3) and liver X receptor were screened and identified. Bioinformatic analysis revealed that these proteins had close relationship with intracellular energy metabolism and translational regulation. Conclusions HSPC016 may regulate the aggregative behavior of DPCs by regulating the levels of intracellular reactive oxygen species (ROS) and interacting with signaling molecules involved in intracellular energy metabolism and translational regulation.

Objective: To evaluate the efficacy of the 45S5 bioactive glass on the reduction of hypersensitivity of teeth prepared for full coverage crowns.Methods: In the study, 31 prepared abutment teeth with vital pulp from 18 patients satisfied the inclusion criteria and exclusion criteria.The patients were informed consent and recruited into the study.This was a before and after self-control, single blind clinical trial.The severity of tooth hypersensitivity at baseline was evaluated, after application of the placebo, which was the powder of zinc oxide, and after application of the 45S5 bioactive glass on the same tooth.Tooth hypersensitivity was evaluated by Yeaple probe and visual analogue scale (VAS) when the tooth was stimulated by a fixed value of mechanical pressure and a blast of cold air respectively.And the dentist gave the subjective score, and the Schiff score at the same time.The three methods of measurement were repeated at baseline, after application of placebo and after application of the powder of 45S5 bioactive glass.Student''s t test and nonparametric test were used for statistical analysis.Results: The indexes measured at baseline, after the application of the placebo and after the application of the 45S5 bioactive glass were as follows, respectively: Tactile value (29.03±9.44) g, (29.68±9.48) g and (44.19±11.19) g, VAS value (50.79±22.92) mm, (46.63±22.06) mm and (30.90±20.30) mm, Schiff score 2.13±0.67, 1.97±0.66 and 1.42±0.56.After the application of bioactive glass, the values of Tactile, the VAS, and the Schiff score were statistically different from those at baseline or after application of the placebo (P<0.05).The data measured at baseline and after application of the placebo were of no statistical difference (P>0.05).Statistical differences between the Schiff scores of the groups were associated with gender, tooth position, primary or secondary tooth preparation.Conclusion: Results of the clinical trial showed that the powders of 45S5 bioactive glass were significantly effective on reducing the hypersensitivity of teeth with vital pulp after the full crown preparation.

Objective　To study the similarities and differences on in vitro replication and expression of hepatitis C virus (HCV) between human fetal hepatocytes (HFH) and 7721 cell line. Methods　Human fetal hepatocytes and a hepatoma cell line 7721 were incubated with a serum from hepatitis C patient. After incubation, the presence of HCV RNA, the expression of HCV NS3 antigens in cells and/or supernatant were examined by RT-PCR, in situ hybridization and immunohistochemistry, respectively. Results　It was found that: ①The intracellular HCV RNA was first detected on d 2～3 post-incubation and then could be intermittently detected in cells and/or supernatant subsequently (HCV RNA could be detected in 7721 cells during a period of at least 66 days. In HFH, HCV RNA could be detected up to 25 days after incubation); ②HCV-NS3 antigen could be expressed in infected cells; ③Minus-strand RNA of HCV was mainly located within cytoplasm by in situ hybridization. Conclusion　The results suggest that both the fetal hpatocytes and the hepatoma cell line 7721 are susceptible to HCV, and especially 7721 cell line can stably support HCV replication in vitro and may be used as the target cell for long-term cultures of HCV.

Objective To construct yeast cDNA expression library of human dermal papillae cells (DPCs) in primary culture.Methods Human dermal papilla cells (DPCs) were isolated by two-step digestion method and cultured in DMEM medium.Total RNA was extracted from primary DPCs that exhibited an aggregative behavior in culture,then,cDNA was synthesized and amplified by using CloneMinerTM cDNA Library Construction kit to construct primary cDNA library and yeast cDNA expression libary.Results The average titer and total clones were 7.0×106 colony forming units(cfu)/ml and 1.4×107 cfu respectively in the primary library,5.5×106 cfu/ml and 1.1×107 cfu respectively in the yeast expression library.The average insert size Was 1.2 kb and the recombination rate was above 95%.Conclusions The yeast cDNA expression library of DPCs in primary culture has been successfully constructed.which will lay a foundation for screening proteins interacting with HSPC016 gene in DPCs with yeast two-hybrid system.

Objective Medication for pituitary adenomas is mainly targeted on the prolactin-secreting and growth-hormone types and shows poor therapeutic effects on other adenomas .Therefore, new drugs urgently need to be developed for this purpose .This study was to investigate the effects of glivec and everolimus on mouse pituitary AtT-20 cells and their molecular mechanisms in vitro. Methods Mouse pituitary AtT-20 cells were incubated with glivec or everolimus or combination of both and their inhibitory effect on the proliferation of the cells was measured by CCK-8 assay.The mRNA levels of AKT and ERK were determined by q-PCR and the ex-pressions of the phosphorylated AKT (p-AKT) and ERK (p-ERK) were detected by Western blot. Results Used alone, both glivec and everolimus inhibited the proliferation of the AtT-20 cells in a time-and dose-dependent manner , but their combination produced a mutually antagonistic effect, with combination index values of 1.13 ±0.06, 1.12 ±0.03, and 1.07 ±0.03 respectively.The two a-gents , either used alone or in combination , induced no significantly inhibitory effects on the mRNA and protein expressions of AKT and ERK ( P >0.05 ).Both glivec and everolimus up-regulated the expressions of p-AKT and p-ERK, and their combination manifested an even stronger effect (P>0.05). Conclusion Both glivec and everolimus inhibit the proliferation of AtT-20 cells when administered alone, but their combination produces an antagonistic effect .Their action mechanism might be that when targeting some signaling path-ways to inhibit cell proliferation , glivec, as well as everolimus , in-duces a feedback activation of AKT and ERK .