Mechanism study of platelet derived growth factor receptor alpha on the bidirectional differentiation regulation of glioma-associated oncogene homolog 1-positive mesenchymal stem cells in mice / 中华口腔医学杂志

Objective: To investigate the role of platelet derived growth factor receptor alpha (PDGFRα) on bidirectional differentiation of glioma-associated oncogene homolog 1-positive mesenchymal stem cells (Gli1+-MSC). Methods: Breeding double reporter transgenic mice ROSAmT/mG/Gli1-CreERt2/PDGFRαfl (Experimental group) and ROSAmT/mG/Gli1-CreERt2 (Control group), 20 mice in each of the two groups at four weeks of age were selected, MSC were isolated from the mouse aortic epithelium. After tamoxifen inducement, the two groups of Gli1+-MSC were screened by green fluorescent protein (GFP) labeling and flow cytometry sorting. PDGFRα was conditionally knocked out in the experimental group, and the control group Gli1+-MSC expressed PDGFRα normally. The two groups of Gli1+-MSC were subjected to adipogenic induction and fibrogenic induction, the Western blotting was performed to detect PDGFRα, adipocyte markers [perilipin and CCAAT/enhancer binding protein alpha (C/EBPα)] and fibrogenic markers [alpha smooth muscle actin (α-SMA) and fibroblast-specific protein 1 (FSP-1)] and semi-quantitative analysis was performed. The degree of cellular adipose differentiation after bidirectional induction of Gli1+-MSC in both groups was observed by oil red O staining and analyzed semi-quantitatively. Results: After tamoxifen induction, Gli1+-MSC could be accurately isolated from flow cytometry by GFP labeling. Via adipogenic differentiation, the expression of PDGFRα in the experimental group (0.017±0.002) was significantly lower than that in the control group (0.184±0.012) (t=25.48，P=0.002). The protein expressions of perilipin (3.138±0.414) and C/EBPα (3.565±0.289) were significantly higher than those in the control group (2.312±0.218 and 2.179±0.103, respectively) (t=6.21，P=0.025；t=6.69，P=0.022). Thus, the knock-out of PDGFRα enhanced the adipogenic differentiation ability of Gli1+-MSC. After fibrogenesis induction, the protein expressions of PDGFRα, α-SMA and FSP-1 in the experimental group (0.030±0.001, 0.932±0.177 and 0.276±0.020, respectively) were significantly lower than those in the control group (0.439±0.006, 1.352±0.170 and 0.835±0.097, respectively) (t=149.40, P<0.001; t=66.38,P<0.001; t=11.41,P<0.08). This suggested that the knock-out of PDGFRα significantly inhibited Gli1+-MSC differentiation toward fibroblasts. After bidirectional induction, significantly less adipocyte formation was seen in the control group and more in the experimental group. Quantitative analysis showed that the amount of oil red O staining in the experimental group (0.461±0.042) was significantly higher than that in the control group (0.017±0.007) after bidirectional induction (t=23.20, P<0.01). Conclusions: PDGFRα plays an important role in the regulation of bidirectional differentiation of vascular adventitial Gli1+-MSC.

Sex determining region Y-box 9 induced microtubule formation and epithelial⁃mesenchymal transition in human oral squamous cell carcinoma CAL27 cells / 华西口腔医学杂志

OBJECTIVES@#This study aimed to explore the effect of sex determining region Y-box 9 (SOX9) on the microtubule formation and epithelial-mesenchymal transition (EMT) of human oral squamous cell carcinoma (OSCC) CAL27 and the underlying mechanism.@*METHODS@#SOX9-shRNA1 and SOX9-shRNA2 were designed and synthesized and then transfected into CAL27 cells. The expression of SOX9 was detected by quantitative real-time polymerase chain reaction. Microtubule formation assay was used to detect the change in the number of microtubule nodules after interfering with SOX9. Immunofluorescence was used to detect the Vimentin content. Western blot was used to detect the protein expression of EMT marker molecules and Wnt/β-catenin pathway-related proteins, such as E-cadherin, N-cadherin, Fibronectin, Wnt, β-catenin, T-cell factor-4 (TCF-4).@*RESULTS@#The expression level of SOX9 significantly decreased after transfection with SOX9-shRNA1 and SOX9-shRNA2 in CAL27 cells (@*CONCLUSIONS@#Interference with SOX9 decreased Vimentin content and inhibited the microtubule formation and protein expression of EMT marker molecules, as well as the expression of proteins related to the Wnt/β-catenin pathway. Thus, SOX9 can induce microtubule formation and EMT in CAL27, which was related to the inhibition of the Wnt/β-catenin pathway activation.

Effects of acupuncture on distribution taxis of paclitaxel in mice with lung cancer / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of acupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) on distribution taxis of paclitaxel in mice with lung cancer to discuss targeted relationship between acupoints and corresponding viscera.</p><p><b>METHODS</b>According to randomized digital table, 315 SPF-grade BALB/C female mice were divided into 7 groups: blank group (group A), model group (group B), medication group (group C), acupuncture at non-acupoint group (group D), acupuncture at Feishu group (group E), acupuncture at Lingtai group (group F) and acupuncture at Feishu and Lingtai group (group G), 45 mice in each one. Except the blank group, the remaining groups were treated with N-nitroso-tris-chloroethyl urea (NTCU) to establish the model of squamous-cell carcinoma. After model establishment, group A, group B and group C were not treated with acupuncture; group A and group B were treated with intraperitoneal injection of 0.9% sodium chlorvde solution by 6 mL/kg while group C was treated with intraperitoneal injection of paclitaxel by 6 mL/kg. The group D, group E, group F and group G were treated with acupuncture at non-acupoint, "Feishu" (BL 13), "Lingtai" (GV 10) and "Feishu" (BL 13) plus "Lingtai" (GV 10), respectively, then were intraperitoneally injected with paclitaxel by 6 mL/kg. The treatment was all given once a day for continuous 10 days. 15 min, 30 min, 1 h, 2 h, 8 h, 12 h and 24 h after the treatments, 6 mice in each group were randomly selected and sacrificed to collect samples of lung, liver, spleen, kidney and heart, etc. High performance liquid chromatography was applied to measure the concentration of paclitaxel in each organ (lung, liver, spleen, kidney and heart) at different time points.</p><p><b>RESULTS</b>(1) The content of paclitaxel in lung, kidney and heart reached the peak at 2 h, then decreased significantly in group C, group D, group E, group F and group G; the content of paclitaxel in spleen showed downtrend at each time point. The content of paclitaxel in liver reached the peak at 2 h in group C and group D; the content of paclitaxel reached the peak at 8 h in group E, group F and group G. (2) The content of paclitaxel in lung in group E and group G was higher than that in group C and group D at each time point (all P < 0.01); the content of paclitaxel in lung in group F was higher than that in group C (P < 0.01) and group D (P < 0.01) only at time point of 2 h. The content of paclitaxel in lung in group G was higher than that in group F at each time point (all P < 0.05). There was no statistical difference between group G and group E (all P > 0.05).</p><p><b>CONCLUSION</b>Acupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) could influ- ence the metabolism of paclitaxel in lung-cancer mice, leading to distribution change in each organ. As a result, it could cause targeting effects, which is more significant at "Feishu" (BL 13) and "Lingtai" (GV 10).</p>

Three-dimensional localization of impacted canines and root resorption assessment using cone beam computed tomography / 华中科技大学学报（医学）（英德文版）

The purpose of this study was to develop a new way to localize the impacted canines from three dimensions and to investigate the root resorption of the adjacent teeth by using cone beam computed tomography (CBCT). Forty-six patients undergoing orthodontic treatments and having impacted canines in Tongji Hospital were examined. The images of CBCT scans were obtained from KaVo 3D exam vision. Angular and linear measurements of the cusp tip and root apex according to the three planes (mid-sagittal, occlusal and frontal) have been taken using the cephalometric tool of the InVivo Dental Anatomage Version 5.1.10. The measurements of the angular and linear coordinates of the maxillary and mandibular canines were obtained. Using this technique the operators could envision the location of the impacted canine according to the three clinical planes. Adjacent teeth root resorption of 28.26 % was in the upper lateral incisors while 17.39% in upper central incisors, but no lower root resorption was found in our samples. Accurate and reliable localization of the impacted canines could be obtained from the novel analysis system, which offers a better surgical and orthodontic treatment for the patients with impacted canines.

Three-dimensional localization of impacted canines and root resorption assessment using cone beam computed tomography / 华中科技大学学报（医学）（英德文版）

The purpose of this study was to develop a new way to localize the impacted canines from three dimensions and to investigate the root resorption of the adjacent teeth by using cone beam computed tomography (CBCT). Forty-six patients undergoing orthodontic treatments and having impacted canines in Tongji Hospital were examined. The images of CBCT scans were obtained from KaVo 3D exam vision. Angular and linear measurements of the cusp tip and root apex according to the three planes (mid-sagittal, occlusal and frontal) have been taken using the cephalometric tool of the InVivo Dental Anatomage Version 5.1.10. The measurements of the angular and linear coordinates of the maxillary and mandibular canines were obtained. Using this technique the operators could envision the location of the impacted canine according to the three clinical planes. Adjacent teeth root resorption of 28.26 % was in the upper lateral incisors while 17.39% in upper central incisors, but no lower root resorption was found in our samples. Accurate and reliable localization of the impacted canines could be obtained from the novel analysis system, which offers a better surgical and orthodontic treatment for the patients with impacted canines.

Application of associated symptom and sign-based acupoint selection in acupuncture-moxibustion treatment / 中国针灸

Based on traditional acupuncture-moxibustion treatment ideas, with differentiation of channels and collaterals as main part and feature, the important role of associated symptom and sign-based acupoint selection in acupuncture-moxibustion treatment is explained from angles of philosophy and medicine. Combined with clinical experience of acupuncture and moxibustion, the category of associated symptom and sign-based acupoint selection is explained in detail to make sure the accuracy of acupuncture-moxibustion differentiation. It could show uniqueness and advantage of theory and clinic in acupuncture-moxibustion and provide theoretical references in making acupuncture-moxibustion prescription to improve effectiveness of clinical treatment.

Microtensile bond strength and morphological evaluations of total-etch and self-etch adhesives to caries-affected dentin / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the microtensile bond strength and bond interface of total-etch or self-etch adhesives to normal dentin and caries-affected dentin.</p><p><b>METHODS</b>A total of 20 molars with occlusal caries lesion were used. The caries-affected dentin was obtained by removing the caries-infected dentin under the guidance of the caries detector. Beyond the level of caries-affected dentin all the enamel and partial dentin were removed. The adhesive systems, two total-etch adhesives (All-Bond 2, Prime&Bond NT) and two self-etch adhesives (Clearfil SE Bond, Xeno III) were applied respectively under the instructions of manufacturers. A block of composite resin was build up superficially. All the teeth were sectioned to obtain bar-shaped specimens with bonded surface area about 0.9 mm x 0.9 mm. The specimens were divided into normal dentin group and caries-affected dentin group via stereomicroscope. The bond strength was tested in a microtensile tester with a crosshead speed of 1 mm/min. The mean values of bond strength were compared using two-way ANOVA. The bonding interface between the dentin and adhesives was qualitatively evaluated under the observation of scanning electron microscope (SEM).</p><p><b>RESULTS</b>Two-way ANOVA revealed a significant influence of both the type of dentin and the adhesive systems tested on microtensile bond strength values. All the adhesives attained higher strength in normal dentin. In normal dentin, there was no significant difference between total-etch and self-etch adhesives. In caries-affected dentin, bond strength of Xeno III was significantly lower than the others. For SEM, the hybrid layer in caries-affected dentin was thicker but more porous than that in normal dentin. Compared with normal dentin, there was fewer resin tag exhibited in caries-affected dentin and no lateral branches were observed.</p><p><b>CONCLUSIONS</b>The total-etch adhesive had higher bond strength than self-etch adhesive systems in caries-affected dentin.</p>

Effects of residual water on microtensile bond strength of one-bottle dentin adhesive systems with different solvent bases / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The wet-bonding technique is recommended for the one-bottle dentin adhesive systems, but the moisture concept varies widely among the instructions of manufacturers as well as among investigators. The aim of this study was to evaluate the effects of different dentin surface moisture on the microtensile bond strength(s) of an ethanol/water-based adhesive system and an acetone-based system to dentin.</p><p><b>METHODS</b>Forty intact human premolars extracted for orthodontic reasons were used. Superficial occlusal flat dentin surfaces of these premolars were exposed, finished with wet 600-grit silicon carbide paper. Under four wet and dry conditions (overwet, blot dry, one-second dry and desiccated), resin composite was bonded to dentin by using Single Bond (SB) or Prime & Bond NT (PB) according to the manufacturers' instructions. The teeth were longitudinally sectioned in the "x" and "y" directions to obtain bonded beams with a cross-sectional area of 0.81 mm(2) with a slow-speed diamond saw. The bonded specimens were tested in tension at a crosshead speed of 1 mm/min until failure of the bonds. Failure modes were observed with a scanning electron microscope. The mean bond strengths were analyzed by one-way ANOVA and Turkey's test.</p><p><b>RESULTS</b>The bond strength of the overwet/SB, blot dry/SB, one-second dry/SB and desiccated/SB groups was 10.87 MPa, 22.47 MPa, 24.91 MPa and 12.99 MPa, respectively. The bond strength of the overwet/PB, blot dry/PB, one-second dry/PB and desiccated/PB groups was 10.02 MPa, 20.67 MPa, 21.82 MPa and 10.09 MPa, respectively. For both SB and PB, the blot dry group and one-second dry group revealed significantly higher bond strengths than the overwet and desiccated groups (P < 0.05).</p><p><b>CONCLUSIONS</b>In order to achieve the highest bond strength to dentin, keeping the dentin surface in an appropriately moist condition is critical for the one-bottle dentin adhesive systems with ethanol/water or acetone solvent.</p>

Microtensile bond strengths of three dentin adhesive systems / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate in vitro the microtensile bond strengths of three dentin adhesive systems and their respective fracture modes.</p><p><b>METHODS</b>A total of 15 intact young human premolars extracted for orthodontic reasons were used. The enamel of occlusal surfaces of these premolar teeth was removed and superficial dentine was exposed, finished with wet 600-grit silicon carbide paper. And then these teeth were randomly divided into three groups. A block of composite resin was bonded respectively with three dentin adhesive systems: All-bond 2 (Group AB(2)), Fluoro-Bond (Group FB) and Xeno III (Group Xeno) according to manufacturers' instructions. The bonded teeth were kept in distilled water for 24 h at 37 degrees C. The roots were removed from the remaining crown approximately 1 - 2 mm below the cemento-enamel junction with a slow-speed diamond saw. The teeth were sectioned to obtain bar-shaped specimens, whose bonded surface areas were about 0.8 mm(2). The specimens were stressed at a crosshead speed of 1 mm/min until rupture of the bond. SEM was used to observe the fracture modes. The mean bond strengths were compared using one-way ANOVA and LSD tests. The frequency of fracture modes was compared using Krukal-Wallis and Mann-Whitney U-test.</p><p><b>RESULTS</b>Mean microtensile bond strengths were (29.56 +/- 5.47) MPa for Group AB(2), (15.81 +/- 7.67) MPa for Group Xeno, and (14.61 +/- 4.50) MPa for Group FB. The bond strength of Group AB(2) was greater than those of the other two groups (P < 0.01). The bond strengths of Group Xeno and Group FB were not significantly different. SEM examination indicated that the adhesive failure was the most mode of fracture.</p><p><b>CONCLUSIONS</b>The microtensil bond strengths of three dentin adhesive systems to normal human dentine were different and the total-etching adhesive All-Bond 2 exhibited the greatest bond strength. It was recommended that dentin adhesive agent should be used according to clinical situation.</p>