Objective: To investigate the effect of human vascular endothelial growth factor on restenosis after angioplasty. Methods: A rabbit model of injured carotid artery was established using percutaneous transluminal angioplasty. The pcDNA3/hVEGF165（500 μg,n=12） and pcDNA3 （500 μg,n=12） were separately transfected into injured arterial wall with 30 min incubation. The carotid artery was imaged by arotic angiography at the end of week 2 and week 4. Pathology analysis and Northern blot analysis were performed for harvested injured artery segment. Results: Arotic angiography showed carotid artery diameter narrowness were obviously lessened at week 2 and week 4 in experimental group than that in control group; H-E stains showed lumina narrow ratio were obviously reduced at week 2 and week 4 in experimental group than that in control group［（9.58±1.35）% vs (31.72±1.72)%；(18.09±2.93)% vs (44.05±3.28)%， P<0.01 ］; By Northern blot analysis, the expression of hVEGF165mRNA in experimental group were upregulated than in contol group. Conclusion: pcDNA3/hVEGF165 can be transfected into smooth muscle cell and continue to secret bioactivity protein at least for 4 weeks; it can accelerate reendothelialization and prevent restenosis.

Objective:To study the mathematical relation between the measurements of root canal curvature obtained by Schneider's method(?_S) and Weine's method(?_W).Methods:The axis of a curved root canal was simplified by an arc (whose radius was r and central angle ?) and 2 tangent line segments (whose lengths were l_1 and l_2),the relation of ?_S and ?_W was studied with the help of mathematical analysis according to the different ways of determining the measured angles on the simplified axis.The graphs of the function were analysed.Results:The following formulas were proved:(1)tan?_S=(1-cos?_W+ ksin?_W)/(sin?_W+kcos?_W),k=l_2/r;(2)?_W/2≦?_S≦?_W.If ?_W was in the interval [0,?],a proximate linear correlation existed between ?_S and ?_W.Conclusion:There is a complex function between the 2 angles(?_S and ?_W) measured by Schneider's method and Weine's method,the ratio of the length of the apical straight part to the radius of the canal curvature is an important factor determining the different values of ?_S and ?_W.

Objective To retrospectively analyze the complications and management of complications during percutaneous implementation of biliary stents in 17 patients. Methods Percutaneous biliary drainage and implementation of biliary stents for malignant biliary obstruction under fluoroscopic guidance was performed in 64 patients. 17 cases developed complications. Bradycardia and hypotension was seen in 5 cases, slipping of the stent into the duodenum in 1, bile duct perforation in 2, the fine guidewire broken in the hepatic parenchyma in 2, malposition of the stent in 3, hepato renal failure leading to hemorrhage in 2, exchanging guidewire broken in the duodenum in 1, and balloon broken leading to stent dislocation in 1, respectively. Results Bradycardia and hypotension in the 5 cases was treated medically and the procedure was interrupted. Slipping of the stent into the duodenum was passed off spontaneously. Broken of the fine guidewire in the liver did not present any complaints until the death of the patients. Perforation of bile duct was treated conservatively with a clinical successful outcome. Malposition of the stent was treated by reimplementation of shorter stents. The broken exchanging guidewire in the duodenum during implementation was removed with the gastroscope. The hepato renal failure leading to hemorrhage had poor prognosis. The broken balloon caused stent dislocation was treated by reimplementation of stent. Conclusion Percutaneous implementation of stent is effective and safe for elderly patients with proximal stenosis of the biliary tract. However, one must be very careful to avoid the complications.

Objective To investigate the expression of Gli1 protein in colorectal cancer (CRC) tissues,and evaluate its association with the prognoses of the patients.Methods A total of 106 patients were enrolled and their clinicopathological characteristics were analyzed.The expression of Gli1 proteins were detected by immunohistochemical staining,and its association with time to recurrence/metastasis (TTR) and overall survival (OS) of the patients were analyzed.Results The positive rate and expression intensity of Gli1 protein in recurrent or metastatic tumor tissues were higher than those in non-recurrent and non-metastatic tumor tissues [86.84 ％ (33/38) vs.58.82 ％ (40/68),1.32 scores vs.0.71 scores,both P ＜ 0.01),while the positive rate and expression intensity of Ki-67 protein remained no difference between both groups [97.37 ％ (37/38) vs.91.18 ％ (62/68),1.89 scores vs.1.75 scores,both P ＞ 0.05).The positive rates and expression intensities of Gli1 and Ki-67 proteins in CRC tissues were higher than those in adjacent tissues [26.42 ％ (28/106) and 0.27 scores;4.72 ％(5/106),0.05 scores] and normal tissues [3.33 ％ (1/106),0.03 scores;0,0.00 scores] (all P ＜ 0.01).Results of univariate analysis showed that the expression of Gli1 protein,tumor grade and lymph node involvement were significantly associated with TTR,but all of the clinicopathological factors had no obvious association with OS.The association remained significant between the expression of Gli1 protein and TTR in multivariate analysis (P ＜ 0.01).Conclusion The expression of Gli1 protein is an independent prognostic marker of recurrence or metastasis in CRC patients,its high expression implicates a high risk of CRC recurrence or metastasis.

Objective To study the clinical and prognostic value of CK19 mRNA-positive circulating tumor cells in early breast cancer patients. Methods We analyzed the peripheral blood in 50 patients with early breast cancer after surgery and before the initiation of any adjuvant treatment for the presence of CK19 mRNA-positive circulating tumor cells using a nest reverse polymerase chain reaction assay. All patients were followed up. Results CK19 mRNA-positive cells were detected in 40.0 %(20/50) of patients with early breast cancer, 12.5 %(3/24) of patients with breast benign lesions, but 5 %(1/20) in healthy individuals (P =0.017,P =0.004); 11 to 20 of them relapsed during the follow-up period (P =0.002). There was no significant association between the detection of CK19 mRNA-positive cell and the patients' menstrual status, tumor stage, tumor size, etc (P ＞0.05). Detection of peripheral-blood CK19 mRNA-positive cells was associated with reduced median relapse-free interval in early breast cancer patients (P =0.007). Conclusion CK19 mRNA is one of the molecular markers for the detection of circulating tumor cells in early breast cancer. Detection of peripheral blood CK19 mRNA-positive cells might be an important predictive value as a marker of relapse in early breast cancer patients.

Objective:To analyze the wall thickness of C-shaped canals in permanent mandibular second molars.Methods:40 ex-tracted permanent mandibular second molars with C-shaped root were collected from native Chinese and were scanned by micro-CT scan-ner.The specimens were reconstructed three-dimensionally by software Mimics 10.01.The roots were sliced from cemento-enamel junc-tion (CEJ)to apex with 1 mm intervals.The cross-sectional root canal configurations were classified into 5 types according to the Fan's Classification.The minimum buccal wall thickness (MBWT),minimum lingual wall thickness(MLWT)and their locations,as well as the wall thickness at the mesial and distal reference sites were detected.Two-way ANOVA and post hoc LSD-t tests were used to com-pare the mean thickness at 4 sides.Results:Among 370 cross-sections of 40 C-shaped roots,C1,C2 and C3 canals were observed on 126(34.1%),46 (12.4%)and 160 (43.2%)sections respectively.Except at the 10 mm level,the mean MLWT was always lower than MBWT;and the MLWT and MBWT were always lower than the wall thickness at the mesial and distal reference sites.The differ-ence was statistically significant (P <0.05)along the root length above the 12 mm level.The MBWT of the C2 and C3 canals was more likely located at the mesial region,and the MBWT of the C1 and the MLWT of C1,C2 and C3 was more frequently located at the center region.Conclusion:The lingual wall of C-shaped canal in mandibualr second molars was the thinnest zone among four sides,and care should be taken during root canal instrumentation and post space preparation to avoid perforation.

Objective To observe the Alberta combined with Infant Language Assessment Scale used in rehabilitation for children with cerebral palsy.Methods From Jun.2012 to Jun.2013,64 cases of cerebral palsy in Children's Hospital Affiliated to Suzhou University were selected according to the different interventions and were randomly divided into an observation group and a control group,32 patients in each group.The control group underwent conventional rehabilitation training included exercise therapy (mainly Bobath therapy,Ueda therapy) and application Infants Language Assessment Scale for treatment,and the observation group on the basis of the method above,set the action in accordance with Alberta infant motor scale(AIMS) assessment to develop rehabilitation programs.Adhere to 3 hours a day of repeated intensive training.Efficacy in children after treatment was compared,and forceful move and fine motor movements and the changes in development quotient(DQ) before and after treatment were compared between 2 groups.Results The total efficiency of the observation group and the control group was 90.6％ and 71.9％,respectively,and there was statistically significant differences between 2 groups (x2 =6.317,P ＜ 0.05).After treatment,the DQ of big movement and fine motor in observation group (47.92 ± 7.15,42.55 ± 8.1 3) were significantly higher than before treatment (36.18 ± 8.23,33.71 ± 10.16) and the control group (38.13 ± 8.21,36.58 ± 8.06),the differences were statistically significant(t =6.235,5.452,6.137,5.243,all P ＜ 0.05).Conclusions The combination of the rehabilitation for children with cerebral palsy in infants language Alberta combined score rating scale,help to improve rehabilitation results and motor function in children,which is of recommendation and application.

Objective: To investigate the dental anatomy of C57BL/6 mice strain. Methods: The skulls and alveolar bones of 16 C57BL/6 mice were scanned by a micro-CT scanner. After 3D reconstruction,the internal and external anatomy of each tooth type was examined. Results: The upper incisors were more curved (mean radius: 2. 37 mm vs. 6. 31 mm) and shorter (mean tooth length: 8. 63 mm vs. 11. 19 mm) than the lower incisors. The palatal roots of the upper 2nd molars (3-rooted),the mesial roots of the lower first molars(double-rooted) and the mesial and distal roots of the lower 2nd molars (double-rooted) had type 1-2 canal form,and all the other roots had a single canal. The mesiodistal diameter of the crown,as well as the numbers of the cusps and developing grooves decreased from the 1st to the 3rd molars. The external tooth shape was very similar as the morphology of the pulp cavity. Conclusion: The C57BL/6 mouse have strong incisors. The tooth size and the anatomic complexity decrease from the 1st to the 3rd molars,which reflects a trend of degradation.

Objective To observe the early effect of Fructus mume extract on KIM-1 and OPN levels in rats with kidney stone formation induced by nanobacteria and to investigate the therapeutic significance of F.mume extract on early kidney stone formation.Methods Nanobacteria were separated and cultured from human upper urinary calculi.The study group appropriately included 54 healthy male SD rats.The renal calculus model was constructed by tail vein injection of nanobacteria.A kidney stone model was created with an F.mume extract intervention,and rats were killed at different stages of the intervention.Real-time polymerase chain reaction was used to detect the KIM-1 mRNA expression in rat renal tissue,and the KIM-1 concentration in urine was detected by using enzyme-linked immunosorbent assay.We detected kidney tissue stone crystals and OPN expression by using hematoxylin-eosin staining and immunohistochemistry.Results Renal tubules of the experimental model were significantly expanded,that is,the formation of renal tubular stones.The early KIM-1 and OPN expression levels were increased.The above-mentioned changes positively correlated with the injection time of the nanobacteria,and F.mume extract antagonized the changes.Conclusion F.mume extract may be useful for the repair of renal injury to reduce kidney stone formation,which may be related to the gene regulation of KIM-1 and OPN.

Objective:To investigate the effects of fluoride exposure on proliferation, apoptosis and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in mice.Methods:BMSCs were isolated and cultured from femur bone marrow of C57BL/6 mice (6 - 8 weeks). The cells in passage 3 were used to detect the surface markers of stem cells by flow cytometry. The cells were cultured in media with a final fluoride concentration of 0.0, 0.1, 1.0, 5.0, 10.0, 15.0, 20.0 and 40.0 mg/L, respectively. The effects of different fluoride concentrations on BMSCs cell proliferation (CCK8 method), apoptosis (flow cytometry analysis), osteogenic differentiation ability [alizarin red and alkaline phosphatase (ALP) staining] were detected. Western blot was applied to detect the levels of apoptosis-related proteins [poly ADP-ribose polymerase (PARP)], mitogen-activated protein kinase (MAPK) pathway member proteins [extracellular regulated protein kinase 1/2 (ERK1/2), c-Jun amino-terminal kinase (JNK), p38 and phosphorylated ERK, JNK, p38 (p-ERK, p-JNK, p-p38)], osteogenic differentiation-related protein [Runt-related transcription factor 2 (Runx2), ALP] and Wnt/β-catenin pathway member proteins [glycogen synthase kinase-3β (GSK3β), phosphorylated GSK3β (p-GSK3β) and β-catenin]. Immunocytofluorescense staining was applied to evaluate the expression levels of p-GSK3β and β-catenin. The two pathways (MAPK and Wnt/β-catenin) were blocked by SP600125 and DKK-1, respectively, to testify their involvement in mechanisms of apoptosis and osteogenic differentiation.Results:The mouse BMSCs were successfully isolated and cultured. Flow cytometry analysis showed that the mesenchymal stem cell surface biomarkers (CD73, CD90 and CD105) were positively expressed. The comparison of cell proliferation at three time points (24, 48 and 72 h) in each concentration group was statistically significant ( F = 65.36, 160.04 and 365.32, P < 0.001), and the comparison of early apoptosis (24 h) in each concentration group was statistically significant ( F = 214.04, P < 0.001); compared with the 0.0 mg/L group, the cell proliferation in 15.0, 20.0 and 40.0 mg/L groups decreased, and the early apoptosis rate in 10.0, 15.0 and 20.0 mg/L groups increased ( P < 0.05). When cells were treated with 15.0 mg/L fluoride for 0 - 24 h, the p-JNK/JNK ratio was higher at 2, 4, 8, 12, 18 and 24 h compared with that at 0 min ( P < 0.05); compared with the fluoride group (15.0 mg/L), the early apoptosis rate of cells after SP600125 block decreased ( P < 0.05), and the protein expression levels of PARP and p-JNK decreased ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, in 0.1 and 1.0 mg/L groups ALP staining was enhanced and the number of calcified nodules increased, and the protein expression levels of Runx2 and ALP in the 0.1 and 1.0 mg/L groups were higher ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, the p-GSK3β/GSK3β ratio and β-catenin protein level were significantly higher in the 0.1 and 1.0 mg/L groups ( P < 0.05); and compared with the fluoride group (1.0 mg/L), addition of DKK-1 significantly decreased the protein expression levels of p-GSK3β and β-catenin and reduced the nuclear entry of β-catenin, and ALP staining decreased and the number of calcified nodules decreased. Conclusions:High concentration of fluoride (> 10.0 mg/L) inhibits the proliferation and promotes apoptosis of BMSCs, while low concentration of fluoride (0.1, 1.0 mg/L) promotes osteogenic differentiation. The MAPK/JNK pathway and the classical Wnt pathway are involved in the above cellular processes, respectively.