Objective To explore the effect of curcumin on the activity and migration of as well as c-kit mRNA expression in melanocytes.Methods Human epidermal melanocytes were isolated from the prepuce in adolescents and subjected to primary culture.To estimate the effect of curcumin on the proliferative activity of melanocytes, some melanocytes were randomly divided into several groups to be cultured in the MelM-2 medium with or without the presence of 5, 10, 20 or 30 μmol/L curcumin, the MelM-2 medium containing curcumin of 5-30 μmol/L served as the drug control groups, and the MelM-2 medium without curcumin served as the blank control group.After 24 and 48 hours of culture, MTS assay was performed to evaluate the proliferative activity of melanocytes.Some cultured melanocytes were randomly divided into 4 groups to be cultured in the MelM-2 medium with 0, 5, 10 and 20 μmol/L curcumin respectively for 48hours.Then, wound scratch assay was conducted to estimate the migratory activity of melanocytes, and real-time fluorescence-based quantitative PCR to quantify the mRNA expression of c-kit in melanocytes.Statistical analysis was carried out by factorial design analysis of variance (ANOVA), one-way ANOVA and least significant difference (LSD)-t test.Results The proliferative activity of melanocytes was significantly decreased at 24 and 48 hours in the 30-μmol/L curcumin group compared with the negative control group (0.783 ± 0.053 vs.1.000 ± 0.018 at 24 hours, 0.637 ± 0.015 vs.0.993 ± 0.064 at 48 hours, both P ＜ 0.05), while no significant differences were observed between the other curcumin groups and the negative control group (all P ＞ 0.05).The 48-hour treatment with curcumin could significantly inhibit the migration of melanocytes in the 5-, 10-and 20-μmol/L curcumin groups compared with the control group (all P ＜ 0.05).The mRNA expression level of c-kit was also significantly reduced at 48 hours in the 5-, 10-and 20-μmol/L curcumin groups compared with the control group (1.799 ± 0.372, 1.539 ± 0.224 and 1.026 ± 0.038 vs.3.371 ± 0.352, all P ＜0.05).Conclusion Curcumin at low concentrations (≤ 20 μmol/L) has no obvious cytotoxicity against melanocytes, but can inhibit the migration of and c-kit mRNA expression in melanocytes, while curcumin at 30 μmol/L can promote the apoptosis of melanocytes.

Objective To evaluate the effect of tacalcitol on the proliferation,adhesion,migration and c-kit mRNA expression of cultured human epidermal melanocytes.Methods Cultured epidermal melanocytes from the prepuce of adolescent males were treated with various concentrations of tacalcitol.Then,cell proliferation was evaluated by tetrazolium salt (XTT) assay after 24,48 and 72 hours of treatment,adhesive activity by using fibronectin-coated culture plates after 72 hours,migratory activity by Transwell assay using a microporous membrane after 24 hours,and the c-kit mRNA expression was semiquantitatively analyzed by reverse transcription PCR after 72 hours of treatment.Statistical analysis was done by repeated-measure analysis of variance and completely random design analysis of variance.Results As repeated-measure analysis of variance showed,tacalcitol of 10-10,10-9,10-8,10-7 and 10-6 mol/L significantly promoted the proliferation of melanocytes (F =9.47,P ＜ 0.01),with significant differences in the promoting effect among various durations of treatment with different concentrations of tacalcitol (F =14.44,P ＜ 0.01),and with significant interaction effect between drug concentration and treatment duration (F =2.47,P ＜ 0.01).The highest proliferation level was observed in melanocytes treated with tacalcitol of 10-s mol/Lfor 72 hours.There was a significant increase in the adhesion rate of human epidermal melanocytes to fibronectin after treatment with tacalcitol of 10-8-10-7 mol/L for 72 hours (both P ＜ 0.01),number of melanocytes migrating through micropore membranes per high-power field (× 200) after treatment with tacalcitol of 10-9-10-8 mol/L for 24 hours (both P ＜ 0.01),and in the c-kit mRNA expression in melanocytes treated with tacalcitol of 10-9-10-7mol/L for 72 hours (all P ＜ 0.01).Conclusion Tacalcitol can promote melanocytes to proliferate,migrate,express c-kit mRNA,and adhere to fibronectin.

A total of 107 vitiligo patients were randomly divided into 3 groups.Group A received an intralesional injection of Bacillus Calmette-Guérin-polysaccharide nucleic acid (BCG-PSN) (n =34),group B Compound Glycyrrhizin Tablets (n =36) and group C both (n =37).Before treatment and 3 months after treatment,cellular immune function was detected for each group.Paired comparisons of 3 groups before and after treatment showed that CD4 +,CD4 +/CD8 + ratio increased (all P ＜ 0.05) and CD8 + decreased (P ＜0.05).After treatment,as compared with groups A and B,CD4 + increases (both P ＜ 0.05) and CD8 + decreased in group C (P ＜0.05).Group C had an efficiency rate of 91.9％ and it was higher than the other two groups (both P ＜ 0.05).An intralesional injection of BCG-PSN plus Compound Glycyrrhizin Tablets could improve immune function and treat vitiligo patients efficiently.

Objective To investigate the effect of hydrogen peroxide (H_2O_2) on apoptosis and calcium ion concentration of skeletal muscle satellite cells (SMSCs) in rats, and to explore the protective effect of erythropoietin (EPO).Methods The cultured SMSCs were divided into five groups: control group,H_2O_2 group, 10, 20 and 40 U/ml EPO intervention groups.Apoptosis rates and calcium ion concentration of SMSCs were analyzed by flow cytometry, and the morphology of apoptotic cells was observed by Hoechst33258 staining.Results The apoptosis rates showed significant differences (all P<0.05) among (1.93±0.57)% in control group, (22.13±1.79)% in H_2O_2 group, (16.47±2.53)%, (4.97±0.55)% and (2.93±0.47)% in 10, 20 and 40 U/ml EPO intervention groups, respectively.And calcium ion concentrations in SMSCs were 12.67 ±0.32, 27.90±0.06 and 44.53±0.93 in 10, 20 and 40 U/ml EPO intervention groups, respectively.There was significant difference in calcium ion concentration between H_2O_2 group and control group (9.70±0.09 vs.51.37± 0.64, P< 0.05).Morphology of apoptosis was observed by Hoeehst33258 dye stains in 10, 20 U/ml EPO intervention group and H_2O_2 group, while there were less apoptotic bodies in 40 U/ml EPO intervention group and control group.Conclusions EPO might have protective effects on SMSCs injured by H_2O_2 through inhibiting apoptosis and calcium ion releasing from SMSCs.

AIM: To investigate the cell cycle arrest ind uced by hypoxia, hypoxia inducible factor-1 and their possible mechanism in huma n ovarian cancer cell line SW626. METHODS: CoCl 2, a chemical inducer of hypoxia and hypoxic cell culture chamber were used to induce chemical and physical hypoxia in human ovar ian cancer cell line SW626. The method of ‘decoy’ was used to block the functi on of HIF-1? because it acts as the core sequence of the target gene as a compe titor combined to the HIF-1?. The cells were divided into group A1 (normal oxyg en), A2 (normal oxygen plus HIF-1? decoy), B1 (CoCl 2), B2 (CoCl 2 plus HIF-1 ? decoy), C1 (hypoxia) and C2 (hypoxia plus HIF-1?). The expression of the HIF -1? protein, mRNA and cell cycle analysis were detected by Western blotting, RT -PCR and flow cytometry (FCM). RESULTS: The expression level of HIF-1? protein in group B1 (3 .75?1.31) and group C1 (3.48?1.01) was significantly higher than that in g roup A1 (0.97?0.31) (P0.05). FCM showed that the G 0/ G 1 phase was markedly increased in group B1 (81.78?24.33) and group C1 (77 .62?22.76) and was significantly higher than that in group A1 (49.49?18.54 ) (P0.05). CONCLUSION: Both CoCl 2 and physical hypoxia could distinctly i nduce cell cycle arrest in G 0/G 1 phase and the expression of HIF-1? in huma n ovarian cancer cell line SW626. HIF-1? plays an important role in cell cycle arrest induced by hypoxia in human ovarian cancer cell line SW626.

Background & Objective: The calcitonin gene-related peptide (CGRP) has a central role in the pathogenesis of migraine, but variations in CGRP-related genes, including the calcitonin gene-related polypeptide-alpha (CALCA) gene and the receptor activity modifying 1 (RAMP1) gene, have not been found to link with migraine in Australian population. The goals of this study were to determine whether variants in the two genes are related to migraine in Chinese population. Methods: Using a case-control approach, rs3781719 and rs145837941 in the CALCA gene and rs3754701 and rs7590387 at the RAMP1 locus was analyzed in a cohort of 504 migraine cases and 529 ethnically matched controls. Genotyping was performed using Sequenom MALDI-TOF mass spectrometry iPLEX platform. Results: The CALCA gene rs145837941 variant was not found in migraine or control group. No significant difference in genotypic and allelic distribution was observed in the other three polymorphisms between migraine cases and controls. All the three SNPs were also not selected as significant factors that independently contributed to susceptibility to migraine in multivariate analysis. In the subgroup analysis, the CALCA rs3781719 seemed to be a significant risk for migraine with aura, but was not statistically significant after FDR correction. Moreover, there was no synergistic relationship between the three SNPs in the multifactor dimensionality reduction analysis for explore locus–locus interactions. Conclusion: Our data suggested that variants in CALCA gene and RAMP1 gene were not associated with migraine in the Han-Chinese population.
Calcitonin Gene-Related Peptide ; Migraine Disorders

Objective To explore the accuracy and clinical efficacy of advanced 3D printing navigation templates in assisting placement of atlantoaxial pedicle screw.Methods A retrospective analysis was carried out on 49 cases of patients with atlanto-axial vertebral fractures and dislocations between June 2013 and June 2016,and all of them were given posterior incision,reduction and internal fixation of atlantoaxial pedicle screw.The patients were divided into advanced 3D printing navigation template group (14 cases),early 3D printing navigation template group (16 cases),and routine pedicle screw placement group (19 cases).Atlantoaxial CT data of patients in advanced 3D printing navigation template group and early 3D printing navigation template group were input into Mimics 17.0,then advanced 3D printing navigation template group and early 3D printing navigation which were used in clinic surgery were designed and printed.The relationship between positions of pedicle screw with the pedicle and bone cortex in plain CT image was observed after operation.The quality of the screw position was assessed and the accuracy of three kinds of screwing methods was compared.The accuracy of the screwing angle was assessed by comparing with the differences between the preoperative designed channel inclination angle and postoperative actual screwing angle.Three groups were compared for differences between operation time,intraoperative blood loss,and scores of cervical nerve scale and visual analogue scale (VAS) of neck and shoulder pain by Japanese Orthopaedic Association (JOA).Results All 49 cases of patients successfully completed the surgery.Patients of the routine pedicle screw placement group,early 3D group and advanced 3D group correspond operation time for 141.2±20.7 min,112.5±12.1 min and 103.1±10.4 min,intraoperative blood loss for 314.0±81.4 ml,243.6±71.2 ml and 181.0+59.1 ml;total accuracy of screwing for 75.0％ (54/72),93.75 ％ (60/64) and 96.43 ％ (54/56).There were statistically significant differences among the routine group,3D group and advanced 3D group in the mentioned programs.There were no statistical differences between advanced 3D group and 3D group in the inclination angle and head tilt angle with the pre-designed values,while there was statistically significant difference between the routine group and the pre-designed value.The accuracy of the inclination angle and bead tilt angle screwing angle were obviously superior in the advanced 3D group and early 3D group to that of the routine group.There were statistically significant differences between preoperative with postoperative VAS scores and JOA scores in the same group,while there were no statistically significant differences among groups in JOA.But there was statistically significant difference between the routine group and the advanced 3D in VAS,and there was no statistically significant difference between the routine pedicle screw placement group and the early 3D in VAS.All three groups of patients had bony fusion of atlantoaxial vertebral body,without loosening,dislocation and fracture of the internal fixators.Conelusion Advanced 3D printing templates in assisting the surgical treatment for atlantoaxial fracture and dislocation can improve the accuracy of pedicle screwing and safety of the surgery,reduce the surgery risk,and obtain satisfied clinical curative effects.

Objective To design and develop intelligent rehabilitation equipment for administering continuous passive motion (CPM) of a rabbit's knee joint after tibial plateau fracture.Methods The equipment constructed had three main parts:the core machinery,electronic control and a control program designed based on bionics principles.Twenty six-month-old New Zealand White male rabbits were randomly divided into sedentary (SED) and CPM groups after their knees had been fractured.The rabbits in the CPM group were given 30 min of early joint rehabilitation once a day for 4 weeks using the CPM equipment,while those in the SED group were kept in their cages and allowed free activity without any special exercise program.The body weight,range of motion and swelling of the affected knee joint were measured before the fracture and on the 3rd,7th,14th,21st and 28th days after the fracture.On the 28th day after the fracture the pathological structure of the articular cartilage on the operative side was observed under a light microscope.Results The equipment ran safely and reliably,and drove the rabbits to move synchronously.It could accurately and conveniently adjust the knee flexion angle,movement speed and movement time.The intelligence of the equipment met the experimental requirements.On the 3rd day after the operation the average range of motion in the joints of both groups had changed significantly compared to that before the fracture.On the 28th day after the fracture the average degree of swelling and range of motion in the CPM group were significantly different from those of the SED group.On the 28th day,deformity and the smoothness of the fracture line in the CPM group were superior to those in the SED group.Moreover,the dominant tissues in the defect area of the CPM group were mainly hyaline cartilage while those in the SED group were mainly repair fibrocartilage.The defect area and its adjacent articular cartilages,chondrocyte regeneration and arrangement,layers of cells and subchondral tidal line recovery of the CPM group were better than in the SED group on average.Conclusion The equipment for knee joint manipulation is convenient to use,reliable and effective for the early rehabilitation of tibial plateau fracture,at least in rabbits.It promotes remodeling of the fracture and cartilage repair after tibial plateau fracture,and also improves range of motion in the knee and reduces swelling.

Objective The purpose of this study was to examine the predictive value of elevated cardiac troponin I (cTnI) and Score for the Targeting of Atrial Fibrillation (STAF) in the diagnosis of cardiogenic cerebral infarction. Methods Two hundred twenty-three patients with acute ischemic stroke were recruited in the study including 38 patients in cardiogenic cerebral infarction (CCI) group and 185 in non-cardiac cerebral infarction (NCCI) group. Clinical data were collected. Chemiluminescence immunoassay was used to detect serum cTnI concentrations in patients and STAF scores were calculated. The clinical baseline data of the two groups were compared. A receiver operating characteristic (ROC) curve was used to determine the boundary value of cTnI and STAF scores in diagnosing CCI and in analyzing their predictive value. Results In the CCI group, the patients were older with higher frequency in atrial fibrillation and ischemic heart disease. Moreover, the NIHSS scores, the value of cTnI and STAF scores were significantly higher in CCI group than in the NCCI group (P＜0.05). The area under the ROC curve of STAF scores was 0.954, and its 95%CI was between 0.924 and 0.985. The area under the ROC curve of the cTnI value was 0.852, and its 95% CI was between 0.788 and 0.916. The cutoff of STAF scores was 4 points, with a sensitivity of 92.1% and a specificity of 89.2%. The cutoff of cTnI value was 0.0085ng/ml, with a sensitivity of 73.7% and a specificity of 84.9%. Conclusion Serum cTnI value and STAF score have a good predictive value for CCI, and STAF score have a higher value than serum cTnI in predicting the diagnosis of CCI. Clinically, serum cTnI and STAF score may be helpful for etiology classification of acute ischemic stroke.