The BRAFV600E mutation maintains the aggressiveness of papillary thyroid cancers requiring downregulation of primary cilia.

Although disorders of primary cilia (PCs) were first reported in human papillary thyroid cancer (PTC) tissues in 1987, their precise role in PTC remains unclear. PCs sense the thyroid follicle colloid environment and act as a cell signaling hub. The present study investigated whether PCs are needed for BRAFV600E-driven PTC. We assessed whether BRAFV600E protein expression correlates with papillary histological architecture and clinicopathological features of PTC. We found that expression of ciliary intraflagellar transport 88 (IFT88) and PC formation were reduced in BRAFV600E-driven PTCs and that loss of cilia may be associated with lymph node metastasis. In PTC cells, the BRAFV600E mutation maintained the aggressiveness of PTC, which was partially related to loss of PCs. Our work confirms that BRAFV600E mutation-driven PC downregulation contributes to maintaining the aggressiveness of PTCs and that manipulating PC can potentially reduce the adverse incidence of PTC in a range of conditions.

Codonopsis pilosula polysaccharides promote osteogenic differentiation and inhibit lipogenic differentiation of rat bone marrow stem cells by activating ß-catenin.

Aberrant bone marrow mesenchymal stem cell (BMSC) lineage differentiation leads to osteoporosis. Codonopsis pilosula polysaccharides (CPPs) have been widely used in traditional Chinese medicines, due to their multiple pharmacological actions. However, little is known regarding their effects on BMSC differentiation. This study aimed to identify the effects and mechanisms of CPPs on osteogenic and adipogenic differentiation in rat BMSCs. An osteoporosis model was established in Sprague-Dawley (SD) rats through bilateral ovariectomy (OVX), and be applied to observe the effect of CPPs on osteoporosis in vivo. The ability of CPPs to affect rBMSC proliferation was determined using the CCK-8 assay, and the osteogenic differentiation of rBMSCs measured by ALP and Alizarin Red S staining. The adipogenic differentiation of rBMSCs was measured by Oil Red O staining. The mRNA and protein levels related to osteogenesis and adipogenic differentiation of rBMSCs were measured using qRT-PCR and western blotting, respectively. Cellular immunofluorescence was used to detect cytokine expression and localisation in rBMSCs. We observed that CPPs ameliorated bone loss in OVX rats. CPPs considerably enhanced osteogenic differentiation by increasing ALP activity and the prevalence of mineralised nodules and promoting the mRNA and protein expression of osteogenic differentiation markers (RUNX2, COL I, ALP, and OPN). Furthermore, it inhibited the accumulation of lipid vesicles in the cytoplasm and the mRNA and protein expression levels of adipogenic differentiation markers (PPARγ and C/EBPα) in a concentration-dependent manner. Meanwhile, CPPs notably increased the mRNA and protein expression of ß-catenin, the core protein of the Wnt/ß-catenin signaling pathway, in a concentration-dependent manner. Adding DKK1, a mature inhibitor of the Wnt/ß-catenin signaling pathway, partially suppressed CPP-stimulated ß-catenin activation, and reversed the acceleration of osteogenic differentiation and the inhibition of lipogenic differentiation. Our observations demonstrated CPPs ameliorate bone loss in OVX rats in vivo, and favour osteogenic differentiation while inhibit adipogenic differentiation of rBMSCs in vitro. The findings suggested that CPPs could serve as functional foods for bone health, and have great potential for the prevention and treatment of osteoporosis.

Biomechanical Analysis of Different Framework Design, Framework Material and Bone Density in the Edentulous Mandible With Fixed Implant-Supported Prostheses: A Three-Dimensional Finite Element Study.

PURPOSE: The objective of this finite element study was to investigate the effect of different framework designs, framework materials, and bone densities on the stress distribution of fixed implant-supported prostheses for edentulous mandibles. MATERIALS AND METHODS: Under the condition of 2-mm cortical bone, 16 models were created in the edentulous mandible to simulate different framework designs (1-piece or 3-piece frameworks) with different framework material (pure titanium, zirconia, polyetheretherketone, or carbon fiber-reinforced polyetheretherketone) in-high or low-density trabecular bone. Then, vertical loading and oblique loading at 75° were applied to the anterior and posterior regions. The stress distribution and stress concentration region of implant and peri-implant bone with different combinations were compared by finite element analysis. RESULTS: The use of the 1-piece zirconia framework in high-density trabecular bone improved stress distribution on implants and peri-implant bone. The region of stress concentration is located in the buccal cervix of the distal implants and the distobuccal portion of the cortical bone in all models. To improve the stress distribution on fixed implant-supported dentures for edentulous mandibles, the 1-piece framework and zirconia represent the better combination. CONCLUSION: Under the condition of 2-mm cortical bone thickness, the full-arch zirconia framework had minimum von Mises stress on implants and peri-implant bone in all models, and high trabecular bone density greatly decreased the stress on cortical bone.

Cardiovascular disease in type 2 diabetes mellitus: progress toward personalized management.

Cardiovascular diseases (CVDs) are the main cause of death among patients with type 2 diabetes mellitus (T2DM), particularly in low- and middle-income countries. To effectively prevent the development of CVDs in T2DM, considerable effort has been made to explore novel preventive approaches, individualized glycemic control and cardiovascular risk management (strict blood pressure and lipid control), together with recently developed glucose-lowering agents and lipid-lowering drugs. This review mainly addresses the important issues affecting the choice of antidiabetic agents and lipid, blood pressure and antiplatelet treatments considering the cardiovascular status of the patient. Finally, we also discuss the changes in therapy principles underlying CVDs in T2DM.

Biomechanical analysis of inclined and cantilever design with different implant framework materials in mandibular complete-arch implant restorations.

STATEMENT OF PROBLEM: Inclined distal implants with posterior framework cantilevers are an alternative to straight implants for the treatment of edentulous jaws, avoiding grafting procedures and utilizing pre-existing bone. However, little is known about the implant, framework, and peri-implant bone stresses exerted by this design. PURPOSE: The purpose of this finite element analysis study was to assess the biomechanical properties of the inclined versus straight design, with different implant framework material to generate implant-supported complete-arch fixed mandibular prostheses. MATERIAL AND METHODS: A finite element model of the edentulous mandible was generated by using 4 implants in 2 distinct configurations: the inclined design and the straight design. Different framework materials were tested: pure titanium, cobalt-chromium alloy, type IV gold alloy, zirconia, polyetheretherketone (PEEK), and carbon fiber-reinforced polyetheretherketone (CFR-PEEK). A 300-N load at a 75-degree angle was applied to the occlusal plane from the lingual side of the buccal cusps of the 2 premolars and the first molar teeth. Subsequently, stresses on the implant, surrounding bone, and framework were measured and analyzed quantitatively and qualitatively. RESULTS: In terms of implant configurations, the inclined design demonstrated less stress on the posterior cortical bone, implants, and framework than the straight design. Comparing the framework materials, zirconia and metal exhibited reduced cortical bone and implants stresses but elevated framework stress when compared with the polymeric frameworks. CONCLUSIONS: From a biomechanical viewpoint, in edentulous patients with excessive posterior alveolar bone resorption, the inclined design exhibited more favorable stress distribution around the posterior implants than the straight design. In implant-supported complete-arch fixed mandibular prostheses, zirconia and metal, particularly cobalt-chromium alloy, distributed the stresses more effectively to the implants and supporting bone than polymeric materials.

The association of obesity with thyroid carcinoma risk.

BACKGROUND: The prevalence of obesity and an increased incidence of thyroid carcinoma (TC) threaten public health in parallel on a global scale. Sufficient evidence supports excess body fatness in thyroid carcinogenesis, and the role and anthropometric markers of obesity have been causally associated with the rising risk of TC. METHODS: A literature search was conducted in PubMed. Studies focused on the effect of obesity in TC. RESULTS: This review mainly discusses the global incidence and prevalence of obesity-related TC. We also review the role of obesity in TC and potential clinical strategies for obesity-related TC. CONCLUSIONS: Excess body fatness in early life and TC survival initiate adverse effects later in life.

MiR-192-5p inhibits proliferation, migration, and invasion in papillary thyroid carcinoma cells by regulation of SH3RF3.

BACKGROUND: The decreased level of miR-192-5p has been reported in several kinds of cancers, including bladder, colon, ovarian, and non-small cell lung cancer. However, the expression and function of miR-192-5p in papillary thyroid carcinoma/cancer (PTC) remains unknown. OBJECTIVE: The present study aimed to explore the function and underlying mechanism of miR-192-5p in PTC development. METHODS: PTC tissues and relative normal controls from PTC patients were collected. qRT-PCR analysis was performed to measure miR-192-5p and SH3RF3 mRNA level in PTC tissues and cell lines. CCK-8 method and FCM assay were used to test cell proliferation and apoptosis in TPC-1 cells, respectively. The abilities of cell migration and invasion were detected by wound healing and transwell assays, respectively. The protein expression was evaluated by Western blot. The interaction between miR-192-5p and Src homology 3 (SH3) domain containing ring finger 3 (SH3RF3) were confirmed by dual-luciferase reporter assay. RESULTS: MiR-192-5p level was obviously decreased in PTC tissues and cell lines. Overexpression of miR-192-5p suppressed proliferation, migration, invasion, and EMT process, while induced apoptosis in TPC-1 cells. In addition, miR-192-5p negatively modulated SH3RF3 expression by binding to its 3'-untranslated region (3'UTR). Silencing SH3RF3 inhibited the migration, invasion, and EMT of TPC-1 cells. In the meantime, matrine, an alkaloid extracted from herb, exerted its anti-cancer effects in PTC cells dependent on increase in miR-192-5p expression and decrease in SH3RF3 expression. CONCLUSION: We firstly declared that miR-192-5p played a tumor suppressive role in PTC via targeting SH3RF3. Moreover, matrine exerted its anti-cancer effects in PTC via regulating miR-192-5p/SH3RF3 pathway.

The Role of Primary Cilia in Thyroid Cancer: From Basic Research to Clinical Applications.

Primary cilia (PC) are microtubule-based organelles that are present on nearly all thyroid follicle cells and play an important role in physiological development and in maintaining the dynamic homeostasis of thyroid follicles. PC are generally lost in many thyroid cancers (TCs), and this loss has been linked to the malignant transformation of thyrocytes, which is regulated by PC-mediated signaling reciprocity between the stroma and cancer cells. Restoring PC on TC cells is a possible promising therapeutic strategy, and the therapeutic response and prognosis of TC are associated with the presence or absence of PC. This review mainly discusses the role of PC in the normal thyroid and TC as well as their potential clinical utility.

miR­875­5p regulates IR and inflammation via targeting TXNRD1 in gestational diabetes rats.

Gestational diabetes mellitus (GDM) is a serious life­threatening disease that affects the mother and fetus. However, the pathogenesis of GDM is still unclear. microRNAs (miRs) play vital roles in the regulation of various cell functions. The present study aimed to investigate the effects of miR­875­5p and thioredoxin reductase 1 cytoplasmic (TXNRD1) in GDM rats and analyze the associated underlying mechanism. A GDM rat model was induced using an intraperitoneal injection of streptozotocin. miR­875­5p knockdown plasmids or TXNRD1 knockdown plasmids were injected into the rats via the caudal vein. miR­875­5p and TXNRD1 expression in the serum were detected using reverse transcription­quantitative PCR (RT­qPCR) or western blot (WB) analyses. The fasting blood­glucose (FBG), fasting serum insulin, triglyceride and high density lipoprotein levels were detected by specific commercial kits. The inflammatory response and the induction of oxidative stress were analyzed by assessing the expression of associated markers via WB, RT­qPCR or commercial kits. The pancreatic and placental injuries were detected by hematoxylin and eosin staining. The results indicated that miR­875­5p expression levels were downregulated, whereas TXNRD1 levels were upregulated in GDM rats compared with normal pregnancy rats. miR­875­5p significantly regulated TXNRD1 expression in GDM rats. miR­875­5p silencing notably reduced FBG and insulin resistance, which was accompanied by reduced expression levels of blood lipid and pro­inflammatory markers as well as reduced oxidative stress. However, the effects of miR­875­5p could be reversed by TXNRD1 silencing. Therefore, the present study indicated that miR­875­5p regulated IR and inflammation by targeting TXNRD1 in GDM rats. miR­875­5p and TXNRD1 may be considered as potential targets for treating GDM.

Evaluation of 1-Piece Versus 3-Piece Framework Designs for the Edentulous Mandible with Fixed Implant-Supported Prostheses: A Clinical, Occlusal and Biomechanical Study.

PURPOSE: This study aims to evaluate the clinical, occlusal and biomechanical performance of 1-piece and 3-piece designs for implant-supported fixed dentures in the edentulous mandible. MATERIALS AND METHODS: A total of 65 patients with edentulous mandibles who underwent fixed implant-supported restorations were recruited and subsequently assigned to 1 of 2 groups based on the framework design (1-piece or 3-piece). The participants underwent clinical and occlusal examination using a periodontal probe, T-Scan III system, and electromyography 12 months after prosthesis delivery. Two mandibular finite element models were created to evaluate stress values and their distribution during function. RESULTS: Ninety-five point four percent (n = 62) of participants in the follow-up period underwent clinical and occlusal examination after prosthesis delivery. Clinical examination revealed a trend towards increased inflammation around the implants in the 1-piece prostheses. Occlusal parameters indicated that the 1-piece design was superior for the masticatory system than the 3-piece design. Biomechanical analysis revealed the highest stress values in the posterior region of the 3-piece design. CONCLUSIONS: On the basis of ease of ensuring oral hygiene, when compared to the 3-piece design, the 1-piece framework design might be the superior therapy for restoring an edentulous mandible, based on occlusal and biomechanical outcomes.

[Gnathology in implant-supported fixed restoration in edentulous mandible].

OBJECTIVE: This study aims to investigate the occlusal and myoelectric characteristics of implant-supported fixed denture in the mandibular region and provide reference for the design of fixed restoration. METHODS: Sixty edentulous patients with implant-supported fixed denture were selected and divided into three groups: group A, 20 cases with implant-supported fixed restoration in the maxillary region; group B, 20 cases with natural dentition, and group C, 20 cases with removable partial denture. The T-scan 8.0 digital occlusion analysis system was used to evaluate the occlusal characteristics of patients in the three groups at intercuspal, protrusion, and left and right lateral positions. Electromyography was used to analyze the myoelectric amplitude and bilateral asymmetry index of the anterior temporalis and masseter of the three groups in different states such as resting and clenching. The relationship between occlusion and myoelectricity was also investigated. RESULTS: In the occlusion analysis by T-scan, the occlusion time, the balance of left and right bite force, the left and right asymmetry of the occlusion center, the trajectory of central occlusion force, and the disclusion time were higher in group C than in groups A and B (P<0.05). No significant differences were observed in the anterior and posterior asymmetry of the occlusion center and percentage of bite force at anterior region among the three groups. In the analysis of myoelectricity, the myoelectric amplitude at resting state and the asymmetry index of masticatory muscles in group C were higher than those in groups A and B (P<0.05). The myoelectric amplitude during clenching in groups A and B groups was higher than that in group C (P<0.05). CONCLUSIONS: In implant-supported fixed restoration at edentulous mandibular, when maxillary includes the removable partial denture, degree of occlusal instability and left and right asymmetry of occlusion center are greater than those with the natural dentition and implant-supported fixed denture at maxillary. The myoelectricity is closely related to occlusion. The removable partial denture can increase the myoelectric activity and reduce the potential of the masticatory muscle. The asymmetry of bilateral myoelectricity is related to the occlusion imbalance.

Metformin promotes osteogenic differentiation and protects against oxidative stress-induced damage in periodontal ligament stem cells via activation of the Akt/Nrf2 signaling pathway.

Periodontal ligament stem cell (PDLSC)-based tissue engineering is an important method for regenerating lost bone in periodontitis. Maintaining or enhancing the osteogenic differentiation of PDLSCs, as well as enhancing the resistance of PDLSCs to oxidative stress, is necessary in this process. As a common hypoglycemic drug, metformin has been reported to have multiple effects on cell functions. This study found that low concentrations of metformin did not affect cell proliferation but did inhibit adipogenic differentiation and promote osteogenic differentiation of PDLSCs. This positive effect was associated with activation of Akt signaling by metformin. Moreover, applying metformin as either a pretreatment or co-treatment could reduce the amount of reactive oxygen species, enhance antioxidant capacity, and rescue the cell viability and osteogenic differentiation that were negatively affected by H2O2-induced oxidative stress in PDLSCs. In addition, metformin was found to activate the Nrf2 signaling pathway in PDLSCs, and knockdown of Nrf2 by siRNA impaired the protective effect of metformin. Taken together, these results indicate that metformin not only promotes osteogenic differentiation of PDLSCs, but also protects PDLSCs against oxidative stress-induced damage, suggesting that metformin could be potentially useful in promoting PDLSC-based bone regeneration in the treatment of periodontitis.

A non-enzymatic glucose sensor with enhanced anti-interference ability based on a MIL-53(NiFe) metal-organic framework.

Non-enzymatic glucose sensors are attracting significant attention owing to their low cost, storage convenience and reusability; however, their poor anti-interference caused by their weak selectivity towards glucose limits their practicability. In the present study, a MIL-53(NiFe) metal-organic framework (MOF) was prepared on Ni foam to serve as a self-supported electrode for non-enzymatic glucose detection. Due to the abundant active sites in the MIL-53(NiFe) MOF and its good stability in an alkaline solution, the sensors exhibited a high sensitivity (41.95 mA mM-1 cm-2) and a low detection limit (0.67 µM). Moreover, the molecular sieve effect of the MIL-53(NiFe) MOF led to a remarkable anti-interference ability, even at the interference concentrations of up to 20% glucose, a higher value than that in human serum. In addition, a heat treatment was carried out to remove the residual terephthalic acid in the MOF tunnels, and this promoted the detection linear range to 2-1600 µM. The reusability, reproducibility and long-term stability of the sensors were also studied, and the results implied good practicability of the MIL-53(NiFe) MOF-based sensors. Furthermore, the good practicability of the sensors was verified by testing human serum samples. The results showed the relative standard deviation of 2.73% from the hospital results, and the standard recovery was nearly 100%.

[Occlusal analysis of implant-supported fixed denture in edentulous patients].

OBJECTIVE: This study aims to evaluate the occlusal characteristics of full edentulous patients with implant-supported prostheses and to provide a reference with the occlusal situation for clinicians. METHODS: A Teetester occlusal analysis system was used with 30 full edentulous patients of implant-supported fixed denture (test group) in comparison with 30 natural dentition (control group). The percentage of occlusal force distribution were measured, as well as the occlusal time at the intercuspal, protrusion, and left and right lateral positions. RESULTS: Compared with control group, the occlusion time, maximum occlusal force in intercuspal of test group significantly reduced (P<0.05); while control group was obviously superior to test group in the left and right bit force degree. Disclusion time in protrusion, occlusion times in lateral positions of test group also significantly reduced (P<0.05). There were no significant differences in average occlusion force, percentage of total force in anterior teeth, and lateral occlusion between test group and control group. CONCLUSIONS: The maximum occlusal force in intercuspal of full edentulous patients with implant-supported prostheses reduce. The occlusal force in protrusive occlusion is concentrated in the front teeth, and the group function occlusion is the main lateral occlusal pattern.

The flavonol glycoside icariin promotes bone formation in growing rats by activating the cAMP signaling pathway in primary cilia of osteoblasts.

Icariin, a prenylated flavonol glycoside isolated from the herb Epimedium, has been considered as a potential alternative therapy for osteoporosis. Previous research has shown that, unlike other flavonoids, icariin is unlikely to act via the estrogen receptor, but its exact mechanism of action is unknown. In this study, using rat calvarial osteoblast culture and rat bone growth models, we demonstrated that icariin promotes bone formation by activating the cAMP/protein kinase A (PKA)/cAMP response element-binding protein (CREB) pathway requiring functional primary cilia of osteoblasts. We found that icariin increases the peak bone mass attained by young rats and promotes the maturation and mineralization of rat calvarial osteoblasts. Icariin activated cAMP/PKA/CREB signaling of the osteoblasts by increasing intracellular cAMP levels and facilitating phosphorylation of both PKA and CREB. Blocking cAMP/PKA/CREB signaling with inhibitors of the cAMP-synthesizing adenylyl cyclase (AC) and PKA inhibitors significantly inhibited the osteogenic effect of icariin in the osteoblasts. Icariin-activated cAMP/PKA/CREB signaling was localized to primary cilia, as indicated by localization of soluble AC and phosphorylated PKA. Furthermore, blocking ciliogenesis via siRNA knockdown of a cilium assembly protein, IFT88, inhibited icariin-induced PKA and CREB phosphorylation and also abolished icariin's osteogenic effect. Finally, several of these outcomes were validated in icariin-treated rats. Together, these results provide new insights into icariin function and its mechanisms of action and strengthen existing ties between cAMP-mediated signaling and osteogenesis.

Multidrug resistance and tumor-initiating capacity of oral cancer stem cells.

PURPOSE: Recent studies in several tumors showed that presence of cancer stem like side population (SP) cells are responsible for chemotherapeutic drugs resistance and tumor relapse. In our present study, we have analyzed the role of SP cells in oral squamous cell carcinoma cell (OSCC) line OSCC-77. METHODS: The oral cancer cell line OSCC-77 was analyzed for the presence of SP cells by FACS using Hoechst 33342 dye exclusion method. Further the FACS-sorted SP and non-SP cells were subjected to drug resistance and sphere formation assays. RESULTS: We identified that the presence of SP cells in OSCC-77 cell line was 3.4%, which was reduced to 0.6% in the presence of verapamil, an inhibitor of ABC transporter. Furthermore, we showed that these SP cells were highly drug-resistant, had increased survival and were highly potent for self-renewal. Also, the clone formation efficiency of SP cells was significantly higher compared to non-SP cells (p<0.01). CONCLUSION: Our data suggest that cancer stem-like SP cells of OSCC-77 cell line contribute to multidrug resistance and are highly involved in tumor relapse. However, further characterization of SP cells at gene expression level and their signaling pathways might provide new insights into the development of novel anticancer drugs.

Sinusoidal electromagnetic fields promote bone formation and inhibit bone resorption in rat femoral tissues in vitro.

Effects of sinusoidal electromagnetic fields (SEMFs) on bone metabolism have not yet been well defined. The present study investigated SEMF effects on bone formation and resorption in rat femur bone tissues in vitro. Cultured femur diaphyseal (cortical bone) and metaphyseal (trabecular bone) tissues were treated with 50 Hz 1.8 mT SEMFs 1.5 h per day for up to 12 days and treatment effects on bone formation and resorption markers and associated gene expression were examined. Treatment with SEMFs caused a significant increase in alkaline phosphatase (ALP) activity and inhibited the tartrate-resistant acid phosphatase (TRACP) activity in the femoral diaphyseal or metaphyseal tissues. SEMFs also significantly increased levels of mRNA expression of osterix (OSX), insulin-like growth factor (IGF-1) and ALP in the bone tissues. SEMF treatment decreased glucose content and increased lactic acid contents in the culture conditioned medium. In addition, treatment with SEMFs decreased mRNA expression levels of bone resorption-related genes TRACP, macrophage colony stimulating factor (M-CSF) and cathepsin K (CTSK) in the cultured bone tissues. In conclusion, the current study demonstrated that treatment with 1.8 mT SEMFs at 1.5 h per day promoted bone formation, increased metabolism and inhibited resorption in both metaphyseal and diaphyseal bone tissues in vitro.

[Inhibition of osthole for resorption of rats femur tissue in vitro].

OBJECTIVE: To investigate osthole effect on femoral tissue resorption activity of rat in vitro. METHODS: Six SD rats weighted (80 ± 5) g were used to isolate and culture femoral tissue (diaphyses and metaphysis) in vitro. The cultured tissue were devided into control group, estradiol group and osthole group. The femoral tissue was treated with final concentration of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol culture in vitro at 48 hours after cultured. Tartrate-resistant acid phosphatase (StrACP) activity, glucose and Lactic acid content, StrACP, MCSF (Macrophage colony stimulating factor) and CTSK (Cathepsin K) mRNA was detected by Real-Time RT-PCR were detected. RESULTS: Concetration of Alkaline phosphatase activity were 2226 and 2498 in 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol respectively. As compared with control group, the activity of StrACP of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol were inhibited at 6, 9, 12 days (P < 0.05); under treatment of in l x 10(-5) mol/L osthole, the content of Lactic acid were increased and the content of glucose were decreased at 3, 6, 9 days (P < 0.05); StrACP, MCSF and CTSK mRNA expression level were inhibited at 6, 9 days (P < 0.05). CONCLUSION: Osthole can inhibit bone resorption and raise the level of nutrition metabolism of femurs tissue.

[Establishment of osteoblast primary cilia model removed by chloral hyrate].

OBJECTIVE: To establish osteoblast model, primary cilla model was removed by chloral hyrate, observe effects of osteoblast primary cilla moved on enhancing ALP staining and calcified nodules staining in electromagnetic field. METHODS: Three 3-day-old male SD rats weighed between 6 and 9 g were killed, cranial osteoblast was drawed and adherencing cultured respectively. Cells were subcultured and randomly divided into 4 groups until reach to fusion states. The four groups included chloral hydrate non-involved group (control group), 2 mM, 4 mM and 8 mM chloral hydrate group, and cultured in 37 °C, 5% CO2 incubator for 72 h. Morphology of primary cilla was observed by laser confocal scanning microscope, and incidence of osteoblast primary cilia was analyzed by Image-Pro Plus 6.0 software. Cells in the correct concentration group which can removed cillia most effectively were selected and divided into 3 groups, including control group (C), Electromagnetic fields group (EMFs), and EMFs with 4 mM chloral hydrate group. DMEM nutrient solution contained 10%FBS were added into three groups and cultured for 9 days and formation of ALP were observed by histochemical staining of alkaline phosphatase. After 12 days' cultivation, formation of mineralization nodes was observed by alizarin red staining. RESULTS: Compared with control group and 2mM chloral hydrate group,4 mM chloral hydrate group could effectively remove osteoblast primary cilla (P<0.01). Removal of osteoblast primary cilla could weaken the formation of ALP and mineralization nodes in osteoblast in EMFS. Compared with EMFs group, the area of ALP and mineralization nodes in EMFs with 4 mM chloral hydrate group were decreased obviously (P<0.01). CONCLUSION: 4mM chloral hydrate could effectively remove osteoblast primary cilia. Primary cilla participate in EMFs promoting formation of ALP and mineralization nodes in osteoblast and provide new ideas for exploring mechanism of EMFs promoting osteoblast maturation and mineralization.