Cryopreservation of calf testicular tissues with knockout serum replacement.

To enrich bovine gonocytes from cryopreserved testicular tissues, the cryoprotection effects of the freezing media containing knockout serum replacement (KSR) were examined. Using Minimum essential medium (MEM) + 10% dimethyl sulfoxide (Me2SO) as the basic medium, calf testicular tissues were cryopreserved in media containing 0, 5, 10, 20, 40, 90% KSR and 5% fetal bovine serum (FBS) respectively. Morphologically, the seminiferous cords and interstitium were well preserved in all groups. The gonocytes were all glial cell line-derived neurotrophic factor (GDNF) family receptor α-1 (GFRα-1) positive. The recovery rates in all KSR groups were higher than that of the 10% Me2SO group, while comparable to the 5% FBS group. The enriched gonocytes expressed gonocyte marker GFRα-1 typically. Collectively, supplementation of 5-10% KSR can achieve comparable cryoprotective effects with using 5% FBS, which is useful in future study due to its defined formulation that is more consistent in quality and stable in supply.

Inhibiting transforming growth factor-ß signaling regulates in vitro maintenance and differentiation of bovine bone marrow mesenchymal stem cells.

Bovine bone marrow mesenchymal stem cells (bBMSC) are potential stem cell source which can be used for multipurpose. However, their application is limited because the in vitro maintenance of these cells is usually accompanied by aging and multipotency losing. Considering transforming growth factor-ß (TGF-ß) pathway inhibitor Repsox is beneficial for cell reprogramming, here we investigated its impacts on the maintenance and differentiation of bBMSC. The bBMSC were enriched and characterized by morphology, immunofluorescent staining, flow cytometry, and multilineage differentiation. The impacts of Repsox on their proliferation, apoptosis, cell cycle, multipotency, and differentiation were examined by Cell Counting Kit-8 (CCK-8), real-time polymerase chain reaction, induced differentiation and specific staining. The results showed that highly purified cluster of diffrentiation 73+ (CD73 + )/CD90 + /CD105 + /CD34 - /CD45 - bBMSC with adipogenic, osteogenic, and chondrogenic differentiation capacities were enriched. Repsox treatments (5 µM, 48 hr) enhanced the messenger RNA mRNA levels of the proliferation gene (telomerase reverse transcriptase [ TERT]; basic fibroblast growth factor [ bFGF]), apoptosis-related gene ( bax and Bcl2), antiapoptosis ratio ( Bcl2/bax), and pluripotency marker gene ( Oct4, Sox2, and Nanog), instead of changing the cell cycle, in bBMSC. Repsox treatments also enhanced the osteogenic differentiation but attenuated the chondrogenic differentiation of bBMSC, concomitant with decreased Smad2 and increased Smad3/4 expressions in TGF-ß pathway. Collectively, inhibiting TGF-ß/Smad signaling by Repsox regulates the in vitro maintenance and differentiation of bBMSC.

Schisandrin A and B enhance the dentate gyrus neurogenesis in mouse hippocampus.

Schisandrin A and B (Sch A and B) are the main effective components of Schisandra chinensis (S. chinensis), which is traditionally used to enhance mental and intellectual functions in eastern Asia. Previously, we reported Sch A and B remarkably affect adult neurogenesis in the subventricular zone of mouse lateral ventricle. Since the neurogenesis in the hippocampal dentate gyrus (DG) is more important to learning, memory and cognition, here we further examined their effects on the adult DG neurogenesis. Phosphohistone H3 (PHH3) immunostaining showed that Sch B significantly enhanced the cell proliferation in the DG. Glial fibrillary acidic protein (GFAP, mostly labels astrocytes and some stem cells) staining was used to further identify the proliferating cell type. Dramatically, increases of GFAP+ cells in both Sch A and B treated groups were observed. What's more, the total numbers of the mature neurons labeled by neuron-specific nuclear protein (NeuN) were also increased in both Sch A and B treated groups compared with the controls. Together, Sch A and B enhance the adult DG neurogenesis by increasing astrocytes/stem cells and improving the survival and maturation of DG neurons. Our study shed a new light on the neuropharmacological functions of the herbal medicine S. chinensis.

[Sutural physical model building in the three-dimensional finite-element model of maxillofacial bones].

OBJECTIVE: To build the physical model of four suturae which are related to the growth of maxilla in the three-dimensional finite-element model of maxillofacial bones. METHODS: A 16 years old volunteer with individual normal occlusion, good periodontium health condition and without diseases of temporomandibular joint was chosen to be the material of modeling. The three-dimensional finite-element model of the volunteer's maxillofacial bones was built using the CT scan and the finite-element modeling method. Finally we built the physical model of four suturae which were related to the growth of maxilla in the model of maxillofacial bones. RESULTS: The model of maxillofacial bones with 86,575 nodes and 485,915 elements was generated. This model contained four suturae including sutura frontomaxillaris, sutura zygomaticomaxillaris, sutura temporozygomatica and sutura pterygopalatine. CONCLUSION: A three-dimensional finite-element model of maxillofacial bones with good biological similarity was developed.

[Quantification of remineralized effect of casein phosphopeptiode-amorphous calcium phosphate on post-orthodontic white spot lesion].

PURPOSE: The aim of this study was to quantify the changes in post-orthodontic demineralized enamel lesion surface areas after using casein phosphopeptide-amorphous calcium phosphate (CPP-ACP). METHODS: Ten orthodontic patients (mean age, 17.7 years) were involved in this study who had multiple decalcified enamel lesion after fixed orthodontic therapy. Ten percent of CPP-ACP paste (GC Tooth Mousse, Recaldent, Europe) was taught to spread on tooth surface two times per day immediately after tooth brush. Standardized intraoral images were taken before and after using it. Image analysis software Image-Pro Plus was used to quantify the visible areas of the demineralized lesions which were expressed as a percentage of total labial surface areas of each tooth. The measurement data before and after using CPP-ACP for 1 month and 2 months were analyzed by paired t test. The measurement of image was repeated again 1 month later to determine the repeatability of the method by calculating the intraclass correlation coefficient between the two results of analysis with SPSS13.0 software package. RESULTS: CPP-ACP significantly reduced the visible enamel demineralization (P<0.01, paired t test). The mean reduction in lesion size after treatment was 4.89%(SD:2.05%; 95% CI: 1.93%-8.77%) for 1 month, and 2 months was 8.359%(SD: 3.472%; 95% CI: 3.76%-15.01%). The intraclass correlation coefficient between the two measurements was 0.968. CONCLUSIONS: CPP-ACP can effectively improve the long-standing postorthodontic demineralized enamel white lesion, and the intraoral image analysis is an reliable and repeatable method to measure white spot lesions. Supported by Research Fund from Bureau of Science and Technology of Changchun City (Grant No. 2008108).

[A three-dimensional finite element study on composite archwire applied in extraction cases].

OBJECTIVE: To explore mechanical properties of the composite archwire (CoAW) applied in extraction cases. METHODS: The mandibular three dimensional model was established by spiral computed tomography scanning and SOLIDWORKS 2001 PLUS software. The archwire model was established by means of ANSYS software. The mechanical analysis and calculation of CoAW were carried out. RESULTS: With the diameter of NiTi part of CoAW gradually augmented (the diameter of stainless steel part kept unchanged), the stress on teeth gradually increased. As the diameter of stainless steel part of CoAW gradually augmented (the diameter of NiTi part kept unchanged), the stress on teeth also gradually increased. The CoAW (0.30 mm NiTi round wire combined with 0.36 mm stainless steel round wire) produced the least force on teeth. There was little contradistinction between six kinds of CoAW and stainless steel wire on the stress of posterior teeth. CONCLUSIONS: The stress on teeth closely correlated with the diameter of the composite archwire.

[Mechanical analysis of composite archwire applied to anterior malocclusion with extraction of bilateral first premolars].

PURPOSE: To optimize the structure of composite archwire applied to anterior malocclusion with extraction of bilateral first premolars. METHODS: The mandibular model was scanned by spiral CT and 30 CT images were obtained, then the mandible finite element model and the archwire model was respectively established by means of SOLIDWORKS software and ANSYS software, mechanics analysis and calculation were carried out. RESULTS: The force produced by composite archwire applied to anteriorly malocclused teeth in the model was greater than by TiNi shape memory alloy but smaller than stainless steel: F(SS)>F(CoAw)>F(NiTi); As the diameter of NiTi in the composite archwire increased (the diameter of stainless steel kept unchanged) , the force on the malocclused teeth gradually increased; As the diameter of stainless steel in the composite archwire increased (the diameter of NiTi kept unchanged),the force on the malocclused teeth increased accordingly. CONCLUSION: The rigidity of the composite archwire increases as the the diameter of NiTi increases; the rigidity of composite archwire also increases as the the diameter of the stainless steel increases.Supported by Research Fund of Jilin Provincial Bureau of Science and Technology (Grant No.20030539-3).