Objective To investigate the effects of fluoride at different concentrations on proliferation and apoptosis of primary rat ameloblast in vitro.Methods Ameloblasts were isolated from tooth germ of 4 days SD rat maxillomandibular molar and cultured in vitro.Cells were treated with NaF at 0.0 (control group),0.4,0.8,1.6,3.2 and 6.4 mmol/L for 24,48 and 72 h,respectively.Inverted microscope was used to observe cell morphology;immunochemistry method was used to identify ameloblasts;3-(4,5-dimethylthiazole-2)-2,5-diphenyl tetrazolium bromide (MTT) assay was applied to measure cell viability at each time point.The cells were treated with 1.6 mmol/L NaF for 24 and 48 h,or after 50 mol/L caspase pan-inhibitor Z-VAD-FMK pretreatment 1 h,1.6 mmol/L NaF treatment for 48 h.Cell apoptosis was then tested by flow cytometry.In addition,activation of caspase-3 and poly (ADP-ribose) polymerase (PARP) were assessed by Western blotting to explore potential involvement of caspase activation in NaF-induced apoptosis.All data analysis was performed using SigmaStat V 3.5 software.Results ①Primary rat ameloblasts were in polygonal shape at low density and appeared like paving stone at high density with obvious nucleus,showing typical morphological characteristics of cells with epithelial origin.②The results of immunochemistry assay indicated that the cultured cells were positive in cytokeratin 14 (CK14) and ameloblastin (AMBN) staining,in accordance with the immunocytochemical characteristics of ameloblasts.③The effects of NaF on ameloblast proliferation were in a dose-and time-dependent manner.For low dose NaF (0.4 and 0.8 mmol/L) groups,cells treated for 24 h had significantly higher cell proliferation rates than that of the control group (0.0 mmol/L,P ＜0.05),the proliferation indexes for 0.0,0.4 and 0.8 mmol/L groups were 1.00 ± 0.00,1.38 ± 0.11 and 1.29 ± 0.13,respectively;the same doses of NaF had no obvious influence on cell proliferation at 48 h (1.00 ± 0.00,1.16 ± 0.14 and 0.94 ± 0.07,P ＞ 0.05);cell proliferation indexes at 72 h were significantly lower than that of the control group (0.87 ± 0.03 and 0.80 ± 0.04,P ＜ 0.05).Medium dose of NaF (1.6 mmol/L) did not cause obvious alterations in cell proliferation at 24 h (0.90 ± 0.08,P ＞ 0.05);while cell proliferation indexes at 48 and 72 h were obviously reduced than that of the control group (0.38 ± 0.03 and 0.26 ± 0.04,P ＜ 0.01).For high NaF concentration (3.2 and 6.4 mmo]/L) groups,cell proliferation indexes were significantly decreased at all time points compared with control cells,the rates for 3.2 mmol/L groups were 0.57 ± 0.14,0.08 ± 0.03 and 0.00 ± 0.00,respectively,and the rates for 6.4 mmol/L groups were 0.11 ± 0.04,0.00 ± 0.00 and 0.00 ± 0.00,respectively (P ＜ 0.01).④Flow cytometry was used to detect apoptosis.The results showed that treatment with 1.6 mmol/L NaF resulted in significantly increased apoptosis in ameloblasts at both 24 h [(5.80 ± 2.03)％] and 48 h [(17.45 ± 4.97)％] compared to the control group [(2.59 ± 0.95)％,P ＜ 0.05].In cells pre-treated with pan-caspase inhibitor Z-VAD-FMK,NaF-induced apoptosis was significantly lower than that of cells treated with only 1.6 mmol/L NaF [(9.43 ± 3.79)％ vs (18.26 ± 3.39)％,P ＜ 0.05].⑤Cleavage of caspase-3 and PARP was detected in ameloblasts treated with 1.6 mmol/L NaF for 48 h.Conclusion Overdose fluoride could inhibit proliferation and induce apoptosis via activation of caspase cascade in primary cultured rat ameloblasts.

Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; surgery ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; diagnostic imaging ; pathology ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed

BACKGROUND:Conventional surgical repair can cause large traumas in patients with knee injuries, and patients often recover slowly after implant fixation, most of whom can appear to have poor recovery of knee function. OBJECTIVE: To explore the folow-up effect of arthroscopic double-steel wire clip fixation on tibial eminence avulsion fracture of anterior cruciate ligament. METHODS: A retrospective analysis was performed on the clinical data of 23 patients with tibial eminence avulsion fractures, who were given arthroscopic double-steel wire clip fixation. The patients were folowed up for 1-6 months. Short- and middle-term therapeutic effect as wel as IKDC and Lysholm scores before and after treatment were observed and analyzed. RESULTS AND CONCLUSION:The operation time was 35-65 minutes, and no complications, such as blood, nerve and anterior cruciate ligament injuries occurred. Moreover, no infection and other poor biocompatible reactions occurred after internation fixation. Al patients were folowed up for 1-6 months. The excelent and good rate was 87% at 1 month after treatment and 96% at 6 months after treatment. Al the patients had improved IKDC score and Lysholm score after treatment (P < 0.05), indicating that the knee function of patients was improved significantly.

Objective To investigate the mechanism of IL-17, the signature cytokine produced by Th17 cells, in OLP lesion. Methods 24 patients with reticular OLP, 19 patients with atrophic-erosive OLP and 13 healthy volunteers were enrolled in this study . Real-time quantitative PCR ( real-time qPCR ) was performed to analyze the expressions of the production of IL-17 and CCL20 mRNA. Results The expressions of IL-17 mRNA in reticular OLP and atrophic-erosive OLP were significant higher than that in healthy oral mucosa (P = 0.0095, P <0.0001, respectively), meanwhile, remarkable increased IL-17 expression in atrophic-erosive OLP group was found compared with reticular OLP group (P = 0.0012). Additionally, the expressions of CCL20 mRNA in reticular OLP and atrophic-erosive OLP were significant higher than that in control group (P=0.0357, P<0.0001, respectively), meanwhile, CCL20 expression in atrophic-erosive OLP was higher than that in reticular OLP. The expressions of CCL20 mRNA rises with the increased expression of IL-17, and were positive correlated with IL-17 expressions in OLP lesions (P=0.003). Conclusions IL-17 production can induce chemokine CCL20 expression in OLP lesion. The signal pathway may promote the migration and infiltration of inflammatory cells in OLP lesions.

Objective:To investigate the polymorphism of HLA-DQA1 genes in Jing nationality of Central Vietnam.Methods:Applied PCR-SSP tecnique to determine the polymorphism of the HLA-DQA1 alleles of 105 healthy children and youth,unrelated individuals in Central of Vietnam.Results:10 HLA-DQA1 alleles were detected of which DQA1*0104 were the most common allele with frequency of 21.3% and lowest frequency is DQA1*0601.Conclusion:The results indicate that HLA-DQA1 alleles polymorphism of Jing nationality in Central Vietnam is different from the other Chinese. [

OBJECTIVETo investigate midterm follow-up results on Asian femoral intramedullary nail in treating segmental and comminuted femoral fractures.

METHODSBetween June 2011 and October 2012,16 patients with segmental and comminuted femoral fractures were treated with minimally invasive reset and Asian femoral intramedullary nail under extension table. Among them, there were 10 males and 6 females aged from 21 to 49 years old with an average of 34.5 years old; the time from injury to operation ranged from 3 to 24 d with an average of 9.1 d. There were 6 cases were type C1,2 cases were type C2 and 8 cases were type C3 according to AO classification. X-ray of femoral segment at 3,6 and 12 months after operation were applied for evaluating fracture healing. Harris score of hip joint and HSS score of knee joint were used to evaluate postoperative function.

RESULTSAll patients were followed up from 24 to 36 months with an average of 28.4 months. Operative time was from 88 to 112 min with an average of 90.7 min; blood loss ranged from 150 to 200 ml with an average of 188.75 ml; the time of fracture healing was from 5 to 9 months with an average of 5.4 months. All incision were healed at stage I. No loosening, breakage of internal fixation and displacement of fracture were occurred. There were no significant differences in Harris score of hip joint at 3, 6 and 12 months after operation (F = 0.07, P = 0.893 > 0.05), 10 cases obtained excellent results, 5 good and 1 moderate. There was no obvious meaning in HSS score of knee joint (F = 0.08,P = 0.876 > 0.05), 9 cases obtained excellent results, 6 good and 1 poor.

CONCLUSIONAsian femoral intramedullary nail could treat segmental and comminuted femoral fractures by using variety of less invasive ways,which has advantages of less trauma, quick recovery of function and satisfied midterm following-up results. But long term following-up effects remains to be seen.

Adult ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Femur ; injuries ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; Fractures, Comminuted ; surgery ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

BACKGROUND:Supination maneuver is mainly used for reduction of radial head subluxation in children, but recently, pronation maneuver has also achieved good results in the treatment of radial head subluxation. OBJECTIVE:To objectively evaluate the efficacy of pronationversus supination maneuvers for the reduction of radial head subluxation by using Meta-analysis. METHODS:PubMed, Cochrane Central Register of Controled Trials (CENTRAL), EMbase, the ISI Web of Knowledge databases, VIP, CNKI, CBM and Wanfang were searched from database establishment to December 2014 for colecting the randomized controled trials (RCTs) about pronationversus supination maneuvers for the reduction of radial head subluxation, and the references of those RCTs were also searched by hand. After study selection, assessment and data extraction were conducted by two reviewers independently. Meta-analyses were performed by using the RevMan 5.2 software. RESULTS AND CONCLUSION:Five studies involving 436 patients were included. The results of Meta-analyses showed that: compared with the supination maneuvers group the pronation maneuvers group had a higher rate of successful reduction at the first attempt [RR=1.17, 95%CI (1.08, 1.28),P=0.000 3] and lower rate of failed reduction [RR=0.25, 95%CI(0.09, 0.65),P=0.005]. There was no significant difference in the rate of successful reduction at the second attempt [RR=1.39, 95%CI (0.75, 2.58),P=0.30]. Based on the results of systemic assessment, the level of evidence assessed by the GRADE system showed that the outcome indicators of the rate of successful reduction at the first attempt and rate of failed reduction were graded as intermediate level; the outcome indicator of the rate of successful reduction at the second attempt was graded as low level. For the poor quality of the original studies, a prudent choice is suggested; and more highly-quality, large-sample studies are needed.

Objective To investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the generation of human myeloid derived suppressor cells (MDSCs) relied on peripheral blood monocytes,and to establish efficient induction system in vitro of MDSCs.Methods Kidney transplantation recipients between January and March 2017 were included in this study.Purified CD14 + cells isolated from peripheral blood were cultured in the presence of GM-CSF with different concentrations for 7 days.Phenotypes and immunosuppressive abilities of induced MDSCs (iMDSCs) were investigated with FACS analyses.Inducible nitric oxide synthase (iNOS) mRNA expression was detected by qRT-PCR to determine the influence of iNOS-pathway on the immunosuppressive abilities of iMDSCs.Results A total of 11 recipients were included in this study.HLA-DR expression decreased sharply after the culture with GM-CSF.iMDSCs showed the similar phenotype characteristics with monocytic-MDSCs (M-MDSCs) as well as significant ability to suppress T cells proliferation and cytokines production.iMDSCs expressed higher levels of iNOS than monocytes,and the inhibitor effects of iMDSCs were significantly reduced after treatment with L-NMMA (1 mmol/L).The variations of phenotype and suppressive ability were concentrationdependent,and more significant changes could be revealed in the group of 10 μg/L GM-CSF.Conclusion GM-CSF-treated peripheral blood monocytes can be efficiently transformed to M-MDSCs,and suppress T cells proliferation and cytokines secretion via iNOS-dependent pathway.These results may contribute to establish MDSCs induction system,which would provide a basis for the clinical application of MDSCs.

BACKGROUND:Percutaneous pedicle screw fixation has been applied in the treatment of thoracolumbar fracture,and has achieved satisfactory clinical efficacy.Injectable calcium sulfate holds good biocompatibility,degradability and fast curing.OBJECTIVE:To explore the clinical efficacy of percutaneous pedicle screw fixation combined with calcium sulfate cement for single-level thoracolumbar fracture.METHODS:Clinical data of 40 patients with single-level thoracolumbar fracture without nerve injury were analyzed retrospectively.All fractured vertebrae were compressed more than 30％ and at least one pedicle was complete.All patients were treated with percutaneous pedicle screw fixation combined with calcium sulfate cement.The Visual Analog Scale and Oswestry Disability Index scores were recorded at baseline,3 months postoperatively and last follow-up;the height of the fractured vertebra body and sagittal Cobb angle were measured on X-ray;the patient's satisfaction and healing rate were recorded.RESULTS AND CONCLUSION:(1) The Oswestry Disability Index at 3 months postoperatively and last follow-up was 16.3％ and 4.4％,respectively.Compared with baseline,the Cobb angle and Visual Analog Scale scores were significantly reduced,and height of the fractured vertebra body was significantly increased after surgery (P ＜ 0.01).(3) The healing rate at last follow-up was 95％,nonunion was not found,and the patients' satisfaction reached 95％.(4) The loss of vertebral height and Cobb angle was found at last follow-up compared with 3 months postoperatively,but had no significant difference (P ＞ 0.05).(5) These results indicate that percutaneous pedicle screw fixation combined with calcium sulfate cement is safe and reliable for single-level thoraclumbar fracture,which not only restores the vertebral height and relieves pain,but also has satisfactory long-term curative efficacy and high healing rate.

Objective To observe the effect of electroacupuncture in preventing delayed cerebral vasospasm (DCVS).Method Sixteen healthy Japanese big ear rabbits were randomized into a treatment group and a control group, 8 in each group. The treatment group was developed into models by intramuscular anesthesia on the 6th day after 5 days’ 20-min daily intervention of transcutaneous acupoint electrical stimulation during a specific time period; the control group was developed into models by the same way on the 6th day but without electrical stimulation. The diameter of basilar artery was measured by using CTA prior to the electrical stimulation and after modeling in both groups, the plasma levels of ET-1, CGRP, and IL-6 were detected, and behavioral changes 1 d, 3 d, and 5 d after modeling were observed.Result After modeling, there was a significant difference in comparing the basilar artery diameter between the two groups (P<0.05). The plasma levels of ET-1, CGRP, and IL-6 in the treatment group were significantly different from that in the control group (P<0.05). In the treatment group, the neural function scores 3 d and 5 d after modeling were markedly different from that 1 d after modeling (P<0.05). There were significant differences in comparing the neural function scores 3 d and 5 d after modeling between the two groups (P<0.05).Conclusion Pre-intervention with electroacupuncture prevents and improves DCVS possibly via modulating the levels of ET-1, CGRP, and IL-6, and subsequently involving in anti-inflammation, immune inhibition, and vasodilation.