ObjectiveTo investigate the role of myocardial calcium-sensing receptor (CaSR) in a rat model of high-level spinal cord injury (SCI).MethodsEighteen healthy male SD rats weighing 250-300 g were randomly divided into 2 groups:sham operation group(group S,n ＝6) and SCI group(n ＝ 12).SCI model was induced by dropping a 10 g weight onto spinal cord (C7) in freely vertical falling along the hollow glass tube from 5 cm height.The blood samples were taken 12 and 24 h after SCI in group SCI and 12 h after SCI in group S,and serum activity of creatine kinase(CK) and MB isoenzyme of creatine kinsse(CK-MB) were measured.Then myocardium specimens were obtained for uhrastructure examination and determination of CaSR mRNA and protien expression by fluorescence quantitative RCR and Western blot.Results Serum activities of CK and CK-MB and CaSR mRNA and protein expression were higher in group SCI than in group S.Serum activity of CK and CaSR mRNA expression were higher,and serum activity of CK-MB was lower at 24 h after SCI than that at 12 h after SCI.There was no significantly difference in CaSR protein expression between the two time points in group SCI.The ultrastructure examination showed that myocardial injury was found in group SCI.ConclusionThe expression of CaSR is up-regulated after SCI in rats,which might be the mechanism of myocardial injury after SCI.

OBJECTIVETo observe the short-term clinical effectiveness of bone ring graft technique and to summarize the key points of related surgical operation to provide comprehensive clinical guidelines.

METHODSFifteen patients with severe alveolar bone absorption were selected to receive bone ring grafting and immediate dental implant. Final fixed prostheses were cemented five months after initial implantation. Cone beam CT scans were conducted on all subjects before the procedure, as well as four months post-operation to evaluate alveolar bone height and level of bone height and absorption around the implants. Four to six months after prosthesis installation, each implant's Jemt classification, gingiva attachment, and probing depth (PD) were analyzed. The difference of PD between implants and adjacent teeth, as well as the difference of the bone absorption between labial and lingual sides, was compared. The survival rate of the bone ring and the retention rate of implants were calculated. Complications and patient satisfaction were also investigated.

RESULTSBone graft survival rate was 94.4% and dental implantation retention rate was 100% four months post-operation. Average bone level increase was (6.06 +/- 1.06) mm, average bone absorption was (1.33 +/- 0.84) mm, and average bone thickness at the neck of the dental implant body was (6.94 +/- 0.73) mm. Approximately 4 to 6 months after crown restoration, average bone level increase was (5.62 +/- 1.03) mm, average bone absorption was (1.51 +/- 1.02) mm, and average bone thickness at the neck of the dental implant body was (6.77 +/- 0.72) mm. The PD around the implant body and the adjacent teeth was statistically insignificant. No major post-operative complication was observed, restorations were successful, and patient satisfaction level was high.

CONCLUSIONBone ring graft technique and immediate dental implantation are relatively simple to perform, and these techniques facilitate reduction in required treatment time. Short-term effect is reliable and satisfactory, whereas long-term outcomes require further follow up and study.

Studying effects of 50 Hz sinusoidal electromagnetic fields (SEMFs) with different intensities on peak bone mass (PBM) of rats may provide a theoretical basis for application of electromagnetic clinical field. 30 female SD rats, 6 weeks of age, were randomly divided into three groups: the control group, 0.1 mT electromagnetic field group (EMFs) and 0.6 mT EMFs. The EMFs groups were treated for 3 h/day. After 8 weeks, we examined their bone mineral densities (BMD) , measured their bone biomechanical properties, and made serum levels of osteocalcin (OC), tartrate-resistant acid phosphatase 5b (TRACP 5b), and histomorphometry. It was found that the BMD (P < 0.01), maximum mechanical load (P < 0.01) in the 0.1 mT group were significantly higher than those in the control group, and Yield strength (P < 0.05), the analyses of serum bone turnover markers and histomorphometric parameters were better than those in the control group (P < 0.05). However, the 0.6 mT group did not have significantly difference comparing with that in the control group. This study proved that 50 Hz 0.1 mT SEMFs can increased BMD, bone strength, and bone tissue microstructure. Therefore, 50 Hz 0.1 mT SEMFs can improve peak bone mass of rats.
Acid Phosphatase ; blood ; Animals ; Bone Density ; Bone and Bones ; physiology ; Electromagnetic Fields ; Female ; Isoenzymes ; blood ; Osteocalcin ; blood ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

This study is to investigate the effect of 8-prenylnaringenin (8-PNG) on osteoclastogensis of bone marrow cells and bone resorption activity of osteoclasts. Osteoclasts were separated from long bone marrow of newborn rabbits and cultured in alpha-MEM containing 10% FBS. 8-PNG was added into culture media at 1 x 10(-7), 1 x 10(-6), 1 x 10(-5) mol xL(-1), separately. 17beta-Estradiol (E2, 1 x 10(-7) mol x L(-7)) was used as positive control. T RAP staining and TRAP activity measurement were performed after 5 days, and the bone resorption pits were analyzed after 7 days. Annexin V staining for the detection of apoptotic osteoclasts was performed after 2, 4, 8, 12, 24, 36 and 48 h separately. The mRNA expression level of TRAP and cathepsin K (CTSK) was measured by real-time RT-PCR. 8-PNG significantly reduced the number of osteoclasts which was TRAP staining positive and with more than three nucleus, the area and number of bone resorption pits decreased obviously in 8-PNG-supplemented groups. The apoptosis rate peaked earlier in the 8-PNG-supplemented groups and the mRNA expression level of TRAP and CTSK decreased significantly. All these inhibitory effects were in a dose dependent manner, the highest effect was obtained by 1 x 10(-5) mol x L(-1) 8-PNG. 8-PNG inhibits bone resorption activity of osteoclasts by inducing osteoclast apoptosis and inhibiting the gene expression and enzyme activity including TRAP and CTSK, and restrains bone marrow cells to osteoclast differentiation.

This study is to investigate effects of genistein on rat femoral bone metabolic in vitro. Rat femoral tissues was isolated and randomly divided into two groups including control group and genistein (1 x 10(-5) mol x(-1)) group. Determinations of alkaline phosphatase (ALP) activity, calcium content and osteoprotegerin (OPG), type I-collagen (Collagen-I), RANKL, Runx-2 and bone morphogenetic protein (BMP-2) mRNA expression were done by real-time PCR. The results showed that 1 x 10(-5) mol x L(-1) genistein could increase the activity of ALP and contents of Ca, regulate bone metabolism activity of OPG, RANKL, BMP-2, Collagen-I and Runx-2 mRNA expression level. Genistein can significantly modulate bone metabolism related gene expression level of rat femoral tissue in vitro, and can increase calcium content and the activity of ALP.

Objective To investigate the effect of creatine phosphate sodium on myocardial protection and calcium-sensitive receptor (CaSR) expression following high-level spinal cord injury.Methods Thirty healthy male SD rats weighing 250-300 g were assigned to sham operation,12-hour injury,24-hour injury,12-hour injury followed by a single intraperitoneal injection of creatine phosphate sodium,and 24-hour injury followed by a single intraperitoneal injection of creatine phosphate sodium according to the random number table,with 6 rats in each group.High-level spinal cord injury was induced at C7 segment by dropping a 10 g weight falling freely along the hollow glass tube from a 5 cm height.Level of blood troponin Ⅰ (cTnⅠ) was measured.Myocardial tissues were collected to study ultrastructure of myocardial cells under transmission electron microscope and CaSR expression using fluorescence quantitative PCR and Western blotting.Results cTnⅠ level was (0.031 ±0.002) U/L and (0.026 ± 0.001) U/L in 12-and 24-hour injury groups,but it was reduced to (0.023 ± 0.002) U/L and (0.018 ± 0.006) U/L at the same time point in treatment groups (P ＜ 0.05).Whereas either in injnry or treatment groups,cTnⅠ level was higher than (0.004 ± 0.002) U/L in sham operation group (P ＜ 0.05).CaSR mRNA level was (0.991 ±0.146) × 10-3 and (1.245 ±0.204) × 10-3 in 12-and 24-hour injury gronp and decreased to (0.880 ± 0.096) × 10-3 and (0.782 ± 0.138) × 10 3 at the same time point in treatment groups (P ＜ 0.05),but all were higher than (0.437 ± 0.065) × 10-3 in sham operation group (P ＜ 0.05).CaSR protein expressed in 12-and 24-hour injury group was (0.627 ±0.066) × 10 3 and (0.809 ±0.154) ×10 3 and lowered to (0.505 ±0.176) × 10-3 and (0.524 ±0.138) × 10-3 at the same time point in treatment groups,but all were higher than (0.331 ± 0.102) × 10-3 in sham operation group (P ＜ 0.05).Transmission electron microscopy demonstrated normal myocardial ultrastructure in sham operation group but impairment in injury groups,but the impairment was significantly improved in treatment groups.Conclusion Creatine phosphate sodium can decrease cTnⅠ level,attenuate the damage to myocardial ultrastructure and down-regulate CaSR after high-level spinal cord injury.

The present research was aimed to investigate the effect of 50Hz, 1. 8mT of sinusoidal electromagnetic fields(SEMFs)on femur tissue cultivation in vitro. The rat femur tissue was isolated from SD rats by method of enzyme digestion, and randomly divided into two groups: SEMFs group and control. The femur tissue of SEMFs groups were exposured under 50Hz 1. 8mT of SEMFs for 1. 5h/time/d, but those in the control groups were without SEMFs treatment. The correlative gene was detected by the Real-time RT-PCR that after SEMFs treatment for 0 (first times treatments is 0 days), 1, 2, 3, 4 and 5d. The alkaline phosphatase (ALP) activity was measured after SEMFs treatment for 3, 6, 9 and 12d respectively. The calcium content was detected after SEMFs treatment for 3, 6, 9 and 12d. The results showed that the OPG and Collagen-1 mRNA expression level was kept at a relatively stable level by SEMFs in the SEMFs group significantly. The Runx-2 mRNA expression level was significantly increased after the SEMFs treatment for 1d and 5d. The ALP activity of femur tissue was significantly increased alter SEMFs treatment for 3d and 9d. The calcium content was higher than untreated groups after SEMFs treatment for 6, 9 and 12d. The SEMFs promoted OPG, Collagen-1, Runx-2 mRNA expression level, ALP activity and calcium content. The result indicated that SEMFs increased the metabolism activity of femur tissues.
Alkaline Phosphatase ; metabolism ; Animals ; Calcium ; metabolism ; Collagen Type I ; genetics ; metabolism ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Electromagnetic Fields ; Female ; Femur ; growth & development ; Male ; Osteoprotegerin ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tissue Culture Techniques ; methods

Objective To explore the role of CT scan for the diagnosis of lung metastasis in stage Ⅲ gestational trophoblastic neoplasia (GTN).Methods To figure out the role of CT scan for lung metastasis in GTN initial diagnosis,treatment and follow-up,93 GTN patients with lung metastasis from January,2015 to December,2016 were retrospectively analyzed in Obstetrics and Gynecology Hospital of Fudan University.Results (1) Among 93 GTN patients with lung metastasis,70 patients with the International Federation of Gynecology and Obstetrics (FIGO) score ≤6 were defined as low risk GTN and 23 patients score score ≥7 were defined as high risk GTN.Forty nine patients had negative chest X-ray findings and 39 cases with pulmonary lesions were identified both by chest X-ray compared to CT scan.Five cases were excluded due to no consensus could make for the results of chest X-ray.The true positive rate of chest X-ray for lung metastasis were 41％ (29/70) in low risk GTN and 43％ (10/23) in high risk GTN patients without statistical difference (x2=0.090,P=0.925).For those patients with positive chest CT scan and negative chest X-ray finding,pulmonary lesions in 32 (65％,32/49) cases were blocked by heart,chest wall or diaphragm in chest X-ray.Seventeen (35％,17/49) patients with lung lesions less than 5 mm had negative chest X-ray results due to the lower sensitivity compared to CT scan.(2) In 88 patients with stage Ⅲ,78 patients had successful initial treatment,but 4 of them were recurrence in twelve months follow-up.Ten patients were chemotherapy resistance for the initial treatment.The initial chemotherapy remission rate in low risk GTN patients was higher than that in high risk ones (x2=4.911,P=0.027).In 49 cases with negative chest X-ray,there was no correlation with the rate of remission,chemotherapy resistance and recurrence in stage Ⅲ patients (P＞0.05).(3) For those patients who had poorly response to initial chemotherapy,the diameters of lesions in lung were unchanged or increased during the treatment,form (5.1±4.1) mm to (7.4±2.8) mm.The pulmonary lesions were continuously shrunk from (7.8 ± 5.3) mm to (4.7 ± 4.4) mm for those patients with complete and partial remission including the recurrent GTN patients (Z=-2.713,P=0.007).Conclusions Patients with GTN in stage Ⅲ have down staging if only use chest X-ray for imaging at the initial diagnosis.Chest CT scan is recommended for primary imaging evaluation of FIGO staging in qualified medical organization.For those patients with persistent abnormal serum hCG level and negative chest X-ray,chest CT scan is strongly recommended to identify the persist or resistant lung lesions and follow up.

Objective:To investigate the role of miR-182-5p regulated primary cilia loss on migration of TPC-1 in papillary thyroid cancers(PTC).Methods:Ten cases of PTC and adjacent tissues were collected from the First Hospital of Lanzhou University, The expression of miR-182-5p in PTC tissue and TPC-1 cells was detected by qPCR, and the frequency of primary cilia was detected by immunofluorescence; Overexpressing miR-182-5p, the migrated number of TPC-1 was detected by Transwell assay; Interfering TPC-1 with siRNA-IFT88, the migrated number of TPC-1 and the frequency of primary cilia were detected, respectively.Results:Compared with control, the expression of miR-182-5p was significantly upregulated in PTC and TPC-1, the frequency of primary cilia in PTC and TPC-1 was downregulated. Overexpressing miR-182-5p increased the migrated number of TPC-1 cell and reduced the number of TPC-1 cell migration(27%, P=0.002); After siRNA-IFT88 treatment, primary cilia in TPC-1 became shorter and thinner, with a decrease in frequency( P=0.001), the migrated number of TPC-1 cell increased, and TPC-1 cell showed smaller nuclei and fewer microvilli. Conclusion:The regulation of primary cilia loss by miR-182-5p through the PI3K pathway contributes to the migration of TPC-1 cells. The loss of primary cilia has an adverse impact on the prognosis of PTC.

OBJECTIVEThe aim of this study is to optimize conditions for labeling human periodontal ligament stem cells (PDLSCs) using enhanced green fluorescent protein (eGFP) infected by lentivirus vector and to obtain PDLSCs with high stable expressed eGFP.

METHODSPDLSCs were transfected with eGFP by lentivirus vector for 48 h via different multiplicity of infection (MOI) (25, 50, 100, 200 and 400) and the infection efficiency were analyzed by both fluorescent microscope and flow cytometry. The proliferation rate of infected PDLSCs was evaluated by MTT. The infected PDLSCs were further for detection of pluripotent, differentiation ability and alkaline phosphatase (ALP) expression ability.

RESULTSThe infection efficiency for each group were 44.7%, 60.9%, 71.7%, 85.8% and 86.9% respectively. Proliferation of PDLSCs was not affected when MOI was below 200; however, at MOI 400, the proliferation ability was affected compared with control group. The pluripotent and ALP abilities of PDLSCs were not changed by the infection.

CONCLUSIONInfection for 48 h at MOI 200 is optimal for labeling PDLSCs with eGFP using lentivirus vector, and the proliferation and differentiation abilities of PDLSCs are not affected obviously.