[Objective] The aim of our study was to seek the mutations in MODY1～5 genes in Chinese population by direct sequencing in probands from families with early-onset type 2 diabetes.[Methods] Variants screening in MODY 1-5 genes were performed by PCR and direct sequencing in 19 probands from early-onset type 2 diabetes families.[Results] We found no mutation but many polymorphisms.There were 6,5,15,1,and 1 variants in MODY 1-5 genes respectively.[Conclusion] Our negative results in MODY genes suggest the genetic heterogeneity of different populations.Mutations in MODY 1-5 genes might not be the cause of diabetes in those 19 families.

Objectives To track bone marrow stem cells (BMSCs) labeled by enhanced green fluorescent protein (EGFP) and superparamagnetic iron oxide ( SPIO ) -poly-L-lysine (PLL) compound by MRI in vitro for autotransplantation into pancreas of type 1 diabetes miniature pigs. Methods The BMSCs were isolated by density gradient centrifugation and attachment culture from type 1 diabetes minipigs' bone marrow. Expressional intensity of EGFP in BMSCs transfected lentivirus-EGFP with a multiplicity of infection Different magnetic resonance scanning protocols were carried out on various density BMSCs labeled by different concentration of SPIO in various time-point in vitro. Results When SPIO concentration was 25mg/L (count in Fe3 + ), the positive Fe3+ -labeling rate of BMSCs was 93. 1%. Most of SPIO particles in BMSCs' cytoplasm were observed in secondary lysosomes, but they were not detected in important organelle as cell nucleus. Comparing with gelatin the MRI of BMSCs labeled with SPIO in the condition with 1 ×104/ml cells density and 25 mg/L Fe3+ concentration in vitro, the signal intensity changes (△SI) after BMSCs labeled with SPIO 3 weeks and 6 weeks in TSE T1WI, TSE T2WI and FLASH T2 * WI sequences were 12%, 41%, 63% and 7%, 28%, 46% respectively (P ＜ 0.01 and P ＜ 0.05, respectively).Conclusions The data showed that the porcine BMSCs labeled with SPIO and EGFP could be traced successfully in vitro by MRI in the suitable sequences.

ovel GCK-E339K mutation might be linked to this MODY2 pedigree.

Objective To research the relationship between free/total prostate specific antigen (f/t PSA), PSA density(PSAD) and prostate cancer(PCa), to explore healthy middle and old-aged,the patients with BPH, the scope of reference value of PSA,frce PSA(fPSA), f/t PSA and PSAD in patients serum with PCa. Methods To de-tect 307 cases of healthy gerontism male,236 of BHP and 41 of PSA and fPSA in patients with PCa,to calcuLate f/t PSA and PSAD, to investigate the dependability between PSA ,I"PSA ,f/t PSA ,P SAD and healthy male, the patients ofBHP,the PCa ,to determine the scope of reference value PSA,fPSA,f/t PSA and PSAD fitting for Chinese. Results PSA and fPSA in patients with PCa are obviously higher than normal control(P <0.01). f/t PSA is obviously lower than the BPH and normal control(P < 0.01). When PSA is 9.25 and f/t PSA is 20%, there are better value of clini-cal diagnosis,when PSAD is 0.18 (AUC = 0.635), we can get best aceuration of diagnosis. Conclusions f/t PSA and PSAD are better than PSA in clinical diagnosis. When PSA is more than 9.25,f/t PSA is less than 20% or PSAD is more than 0.18 is significance for diagnosis. The normal reference value of PSA less than 9.25,f/t PSA more than 20%, PSAD less than 0.18 is fit for Chinese.

Objective:To evaluate the clinical significance of the coincidence test with 3D mirror image of unaffected side in the treatment of contralateral calcaneal fracture.Methods:A retrospective analysis was performed of the 35 patients who had been treated for single calcaneal fracture by coincidence test with 3D mirror image of unaffected side from January 2016 to June 2018 at Department of Trauma Orthopedics, The Sixth Hospital of Ningbo. They were 29 males and 6 females with an average age of 44 years (from 18 to 71 years). By Sanders classification, 19 cases were type Ⅱ and 16 cases type Ⅲ. CT scan and reconstruction of both calcaneuses was performed. The image of unaffected calcaneus was mirrored by Mimics software to guide virtual treatment of contralateral calcaneal fracture. Preoperatively the virtual bilateral calcaneuses and the reduced calcaneus were 3D printed to guide the actual operation. The coincidence test with 3D mirror image of unaffected side was used to evaluate fracture reduction after operation. Postoperative parameters such as B?hler Angle, Gissane Angle, calcaneal width, and coincidence of the affected calcaneal bone were recorded at 12 months after surgery. The clinical outcomes were assessed by American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and visual analogue scale (VAS) score.Results:In this cohort, operation time averaged 43.9 min (from 30 to 55 min), time from admission to operation 3.6 d (from 0 to 14 d), and hospital stay 6.3 d (from 3 to 11 d). The patients were followed up for an average of 18.8 months (from 13 to 24 months). All patients achieved bone union after an average of 17 weeks (from 12 to 26 weeks). Re-collapse of subtalar articular surface, talocalcaneal joint fusion, loss of calcaneal height or length, and calcaneal width increase were observed in none of the patients. The VAS scores averaged 2 (from 0 to 4), and the AOFAS scores 93.3 (from 76 to 98). The total coincidence was 95.2%±3.5%. Spearman test indicated a positive correlation between postoperative total coincidence and AOFAS score ( ρ=0.588, P<0.05). Conclusion:The coincidence test with 3D mirror image of unaffected side can be used to treat contralateral calcaneal fracture, with advantages of reduced surgical time, increased surgical accuracy and fine clinical outcomes.

Objective:To compare the biomechanical characteristics of screw only spatial weaving fixation and calcaneal plate fixation in calcaneal fractures.Methods:Sanders type III calcaneus fracture model was established by using calcaneus model specimens: the physiological model group were the normal calcaneal models; the steel plate group were conventional steel plate fixation fracture models; the metal screw group were fracture models with only metal screw weaving fixation; seven and nine absorbable screw spatial weaving groups (seven absorbable screw group, nine absorbable screw group) were used to weave and fix fracture models with seven and nine absorbable screws. Cyclic test and mechanical compression test were carried out, and load-displacement curves were recorded. The material properties of metal screw spatial weaving and calcaneal anatomical plate system were replicated, finite element fracture models were established, and the calcaneal internal fixation models of plate screw group and spatial weaving screw group were completed by reverse processing. The changes of biomechanical characteristics of calcaneal bone in human (70 kg) standing on one foot were simulated, and the distribution of structural strength was analyzed by Von Mises equivalent stress cloud diagram and displacement cloud diagram.Results:In the cyclic test of 20-200 N load, the physiological model group, the plate group, the metal screw group, the absorbable 7 screw group, the absorbable 9 screw group were 0.87±0.22, 0.82±0.08, 0.70±0.12, 1.04±0.13 and 0.83±1.76 mm, the difference in model gap was statistically significant ( F=3.16, P=0.037). Among them, the absorbable 7 screws group was larger than the metal screws group ( t=4.28, P=0.003), and the other pin-two comparisons were not statistically significant ( P>0.05). The deformation of the five groups was 0.37±0.06, 0.38±0.07, 0.38±0.06, 0.52±0.07 and 0.42±0.07 mm, and the difference was statistically significant ( F=4.39, P=0.010). The deformation of absorbable 7 screws group was greater than that of physiological model group, the plate group and metal screw group ( t=3.69, P=0.006; t=3.25, P=0.012; t=3.51, P=0.008). In static test, compression displacement was 7.14±0.79, 7.30±0.66, 6.95±0.28, 8.19±0.61 and 7.16±0.55 mm, the difference was statistically significant ( F=3.28, P=0.032). The displacement of the absorbable 7 screws group was greater than that of the metal screws group ( t=4.13, P=0.003). The stiffness changes were 570.60±122.62, 512.86±80.77, 497.40±66.50, 456.21±58.19 and 560.39±94.40 N/mm, respectively, with no statistical significance ( F=1.44, P=0.258). The results of finite element analysis showed that under 3 500 N axial pressure load, the maximum compression displacement and stiffness of the plate and screw set were 6.47 mm, 540.96 N/mm, and the Von Mises equivalent stress peaks were 450.31 and 353.15 MPa, respectively. The maximum compression displacement and stiffness of the braided screw group were 5.25 mm, 666.67 N/mm, and the peak Von Mises equivalent stress of the screw was 396.20 MPa. Conclusion:Compared with lateral plate fixation, spatial weaving fixation can provide sufficient biomechanical stability for calcaneal healing and is superior to plate fixation in terms of structural stability, which may help to improve the effectiveness of calcaneal fracture fixation.

Objective:To study a safe anatomical range of the lateral calcaneus for safe surgical approaches.Methods:In 15 cadaveric specimens of the calcaneus, the coordinate axes X, Y, Z were established on the apex of the fibula (the lowest point) as the origin on the lateral side of the calcaneus. In the main part of the lateral calcaneus in the third quadrant of coordinates, the spatial quantification was conducted of the 4 important anatomical structures on the lateral calcaneus (calcaneofibular ligament insertion, common peroneal tendon sheath, lateral calcaneal artery and nerve). The trend trajectory of each structure was summarized. Based on the summary of above measurements, the safety ranges were coincided to find a safe anatomical range for a surgical approach that might lead to the least probability of damaging the important lateral calcaneal structures.Results:The distances from the 4 important anatomical structures to the X, Y, and Z axes of the main part of the lateral calcaneus in the third quadrant are respectively: (21.40±3.38) mm, (18.47±3.91) mm, and (25.06±3.45) mm for the lateral calcaneal artery; (16.53±4.77) mm, (16.27±3.68) mm, and (23.13±4.00) mm for the lateral calcaneal nerve; (9.73±1.73) mm, (11.47±2.13) mm, and (10.87±1.59) mm for the common peroneal tendon sheath; (22.33±2.84) mm for the calcaneofibular ligament insertion. The above 4 important structures mainly converge at the origin O and the anterior 1/3 of the tangent to the outer edge of the calcaneus. In this range, a safe surgical approach can be designed that is located at >2.5 cm below the lowest point of the fibula and 2.1 cm behind the rear side of the arc range.Conclusion:A safe surgical approach can be designed in the safe convergence range of important anatomical structures of the lateral calcaneus to reduce iatrogenic injury to the important structures on the lateral side of the calcaneus.

[Abstract] Objective: To screen candidate epitopes of breast cancer HLA-A2 restrictive neoantigen and to identify high frequency mutation sites in breast cancer neoantigen by using bioinformatics method. Methods: NCBI and GDC databases were used to search missense mutation sites formed by single nucleotide mutation in breast cancer among reported literatures and sequencing data. The new antigen epitopes were predicted by HLA-A2 antigen epitope prediction website BIMAS, SYFPEITHI and artificial neural networkbased NetMHC4.0, and the epitopes with TAP binding power less than Intermediate were eliminated. The candidate epitopes were prioritized by mutation frequency and prediction results. Results: A total of 17 high-frequency mutation genes, including BTLA, ERBB2 and NBPF12 etc, were screened by the above-mentioned methods, and a total of 26 neoantigen epitopes were identified. The binding power of epitopes predicted using BIMAS and SYFPEITHI showed great difference (P<0.05), epitopes in high priority as GSTP1 (A114V , mutation frequency of 5.94%) and BRCA2 (N991H, mutation frequency of 5.40%) etc, were expected to be candidate neo-antigen epitopes; however, their mutation frequency was relatively too low to achieve“universal use” . The possibility of these epitopes used as general breast cancer neo-antigen epitopes is less likely. Conclusion: The common mutation frequency of breast cancer is lower than that of other tumors; it ’s difficult to find“universal”new antigen epitopes of breast cancer; the individualized neoantigen vaccine may be of more promise, which needs further research.