Objective To investigate the effect of antisense oligonucleofide(PCNA ASON)and vascuiar endothelial growth factor (VEGF) gene traneduction on restenosis after balloon angiopasty.Memods chinese rabbits were randomly divided into control group(I),PCNA ASON(II),VEGF only (Ⅲ),PCNA ASON+VEGF(Ⅳ)groups.Each group included 7 rabbits.Restenosis wasevaluated by pathology immunohistochemistry and Western-blotting for the expression of PCNA,and the depth and area oftunica media and tunicca intima were measured. Results All rabbits experienced restenosis on different severities,especially in control group.Lesions were less severe in PCNA ASON and VEGF groups than in controls.The proliferation of smooth muscle and intima significantly ameliorated in PCNA ASON and VEGF combination group that in PCNA ASON or VEGF groups(P<0.01).But the difference between the PCNA ASON and VEGF group was not significant(P>0.7). Conclusion PCNA ASON and VEGF gene transductionn are effective in preventing restenosis after balloon angioplasty caused vessel injury in rabbits.

Objective To investigate the effect of triptolide (TP) on the expression of hypoxia inducible factor 1 alpha (HIF1α) and vascular endothelial growth factor (VEGF) in the human umbilical vein endothelial cells (HUVECs) under hypoxia. Methods (1) HUVECs were treated with 0, 40, 80, 160 and 320 nmol/L TP (named with hypoxia group, TP40 group, TP80 group, TP160 group and TP320 group, respectively) under the hypoxic condition (37℃, 5%CO2, 1%O2, 94%N2) for culturing 12 hours. Meanwhile, cells cultured under normoxia condition (without TP added) were set as the normoxia group. Western blot assay was used to detect the expression of HIF1αin each group. (2) The cells were divided into normal control group, hypoxia group and TP80 group. The immunofluorescence method was performed to detect the localization of HIF1α in cells. (3) Expressions of VEGF were detected by Western blot assay in TP80 group and hypoxia group. (4) The cells were divided into hypoxia group, TP80 group, TP80+KF20 group (80 nmol/L TP and 20μmol/L KC7F2), and TP80+KF30 group (80 nmol/L TP and 30 μmol/L KC7F2). After 12-hour culturing, Western blot assay was used to detect the expressions of HIF1α and VEGF in each group. Results (1) Under the normoxia condition, no HIF1α was detected in HUVECs. The expression level of HIF1αwas significantly increased in TP80 group than that in hypoxia group (P<0.05), while there was no significant change in expression of hypoxia HIF1αin TP160 group and TP320 group compared with that of hypoxic group. (2) The immunofluorescence result showed that HIF1α was mainly expressed in the nucleus. (3) The expression of VEGF was significantly increased in TP80 group than that in hypoxia group (P < 0.05). (4) After the intervention of KC7F2, HIF1αand VEGF expression levels were significantly decreased in the TP80+KF20 group and the TP80+KF30 group than those in the TP80 group (P<0.05). Conclusion TP can improve the expression of HIF1αand VEGF to accelerate the proliferation of endothelial cells under hypoxia condition.

Objective To investigate the neuroprotective effect and its mechanism of local hypothermia combined with nicotinamide phosphoribosyltransferase (NAMPT) on cerebral ischemia-reperfusion injury in rats.Methods Fifty-four Sprague-Dawley rats were randomly divided into sham operation group,model group,NAMPT group,local hypothermia group,and combined treatment group (NAMPT + local hypothermia).A rat model of local cerebral ischemia-reperfusion was induced by suture method.Infarct volume and cerebral edema volume were assessed by 2,3,5-triphenyltetrazolium chloride staining after 2 h cerebral ischemia and 24 h reperfusion in rats.Evans blue staining was used to assess the extent of blood-brain barrier damage,and a 12-point scale was used to assess neurological deficits.Results The infarct volume in the local hypothermia group,NAMPT group,and combination treatment group was significantly lower than that in the model group (all P ＜0.05).The infarct volume in the combination treatment group was significantly lower than that in the NAMPT group (P ＜0.05).The infarct volume in the combination treatment group was lower than that in the local hypothermia group,but it did not reach statistical significance.The neurological function scores of the local hypothermia group,NAMPT group,and combination treatment group were significantly lower than those of the model group (all P ＜0.05).The score of the combined treatment group was significantly lower than that of the NAMPT group and the local hypothermia group (all P＜0.05).Evans blue leakage in the local hypothermia group,NAMPT group,and combination treatment group was lower than that in the model group (all P ＜0.05),but the differences between each treatment group were not statistically significant.Conclusion NAMPT and local hypothermia combination therapy showed better neuroprotective effects on cerebral ischemia-reperfusion injury,suggesting that the combination therapy had clinical transformation prospects.

The development of an expert consensus based on specific domestic situations will provide practical guidance to the efforts aiming at improving cleft care in China. The team approach of twenty-one cleft centers were pooled together, covering pre-surgical orthopedics, primary surgical repair, orthodontic treatment, alveolar bone graft, secondary deformity correction, palatal fistulae repair, the diagnosis and treatment of velopharyngeal incompetence, speech therapy, otitis media management, and skeletal deformity correction. Agreement was achieved among the authors concerning the application of critical surgical and non-surgical techniques. The ambition of this consensus is to introduce more clinicians to the revolution of sequential treatment of clefts, and form the basis for a more comprehensive cleft care manual in the future.
Alveolar Bone Grafting ; Cleft Lip ; Humans ; Velopharyngeal Insufficiency

Objective:To explore the spatial support capacity and its influence in osteogenic effect of composite biofilm based on poly(propylene carbonate)(PPC)/poly(butylene succinate)(PBS)in rabbit tibial bone defect models,and to clarify its barrier functional reliability and osteogenetic effect in vivo.Methods:The composite biofilms of PPC/PBS and PPC/PBS/collegen type Ⅰ(Col-Ⅰ)(PPC/PBS/Co)were prepared.Eighteen Japanese big-ear rabbits were selected and two bone defects were prepared on each side of the tibia of the rabbits.Six rabbits were randomly selected to place PPC/PBS composite biofilm on the bone defects,2 rabbits were executed at 2,4,8 and 12 weeks after operation,and the surface microstructures of PPC/PBS composite biofilm in the rabhit bone defect area were observed by scanning electron microscope(SEM).The experiment was divided into blank control group,PPC/PBS composite biofilm group,BME-10X collagen membrane group,and PPC/PBS/Co composite biofilm group.The above biofilms were placed on the corresponding bone defects of rabbits by operation,while no biofilm was placed in the rabbits in blank control group.Three rabbits were killed at 2,4,8 and 12 weeks after operation respectively,and the gray values of regenerated bone in the bone defect areas of the rabbits in varrous groups were detected by soft X-ray;the fluorescence intensities of regenerated bone tissue in the bone defect areas of the rabbits in various groups were observed by laser scanning confocal microscope after fluorescence labeling.The pathomorphology of regenerated bone tissue in the bone defect areas of the rabbits in various groups were observed by HE staining and modified Gomori staining,and the expression levels of bone morphogenetic protein 2(BMP-2)and osteopontin(OPN)in the regenerated bone tissue in bone defect areas of the rabbits in various groups were detected by immunohistochemical staining.Results:In general,the PPC/PBS composite biofilm was tightly covered in the bone defect area without displacement and collapse.The SEM results showed that the porous surface of PPC/PBS composite biofilm appeared micropore structure and the number of micropores was increased with the prolongation of time,while the smooth surface of biofilm basically did not form the micropore-like structure.The results of soft X-ray detection showed that the gray values of regenerated bone tissue in bone defect areas of the rabbits in various groups were increased with the prolongation of time,and the gray value of regenerated bone tissue in bone defect areas of the rabbits in PPC/PBS/Co composite biofilm group was significantly higher than those in other groups(P＜0.05).The confocal micrscope results showed that the fluorescence intensity of regenerated bone tissue in bone defect areas of the rabbits in PPC/PBS/Co composite biofilm group was similar to those in blank control group at 4,8,and 12 weeks;compared with PPC/PBS composite biofilm group and BME-10X collagen membrane group,the fluoresence intensity of regenerated bone tissue in bone defect areas of the rabbits in PPC/PB/Co composite biofilm group at 4 weeks was increased(P＜0.05),and the fluoresence intensity of regenerated bone tissue in bone defect areas of the rabbits at 8 and 12 weeks were decreased(P＜0.05).The results of HE staining and modified Gomori staining showed that compared with PPC/PBS composite biofilm group and BME-10X collagen membrane group,the new bone formed faster in PPC/PBS/Co composite biofilm group and blank control group at 2 and 4 weeks,and the lamellar bone mineralization was higher at 12 weeks.The immunohistochemical staining results showed that compared with blank control group,PPC/PBS composite biofilm group and BME-10X collagen membrane group,the expression levels of BMP-2 and OPN proteins in the regenerated bone tissue in bone defect areas of the rabbits in PPC/PBS/Co composite biofilm group at 2 and 4 weeks were increased(P＜0.05 or P＜0.01);compared with blank control group and PPC/PBS composite biofilm group,the expression levels of BMP-2 and OPN proteins in the regenerated bone tissue in bone defect areas of the rabbits in BME-10X collage membrane group were decreased(P＜0.05 or P＜0.01).Conclusion:PPC/PBS composite biofilm has excellent spatial support capacity and reliable physical barrier function.The PPC/PBS/Co composite biofilm has a good effect in guiding bone regeneration in vivo.

Objective: To explore the effect and mechanism of chrysin and chloroquine(CQ) on the proliferation and apoptosis of ovarian cancer cells. Methods: Ovarian cancer specimens and normal ovarian specimens were selected after surgical resection and immunohistochemical experimental methods was used to detect the expression level of autophagy-related protein LC3 in ovarian cancer and normal ovarian tissues. The CCK-8 method was used to detect the proliferation of ovarian cancer cell SKOV3. Western blot and immunofluorescence were used to detect the expression of LC3 in SKOV3 cells. An electron microscope was used to detect the number of autophagosomes in SKOV3 cells. Flow cytometry was used to detect SKOV3 cell apoptosis. Results: The expression level of LC3 in ovarian cancer was higher than that in normal ovarian tissues. SKOV3 cells were treated with chrysin at concentrations of 0, 20, 40, 60, 80, 100 μmol/L for 24 hours. As the concentration increased, cell proliferation decreased significantly. Compared with the control group, 40 μmol/L, chrysin significantly up-regulated the expression of LC3 after 24 hours(P<0.05). Compared with the control group, after 24 hours of SKOV3 cells treated with chrysanthemum, more autophagosomes were observed under transmission electron microscope(P<0.05), but no autophagosome was observed in the control group. When chrysin and chloroquine were used in combination for 24 hours, the expression of LC3 protein in the combined treatment group was higher than that in the chrysin and CQ treatment group(P<0.05). Chrysin could obviously induce apoptosis of SKOV3 cells, and the group of Chrysin+CQ showed the highest proportion of apoptosis(P<0.05). Conclusion Chrysin can induce autophagy to inhibit the proliferation of ovarian cancer cells and promote apoptosis.