Objective To observe and identify the osteogenic activity,biocompatibility and mechanical property of a type of macro-pore bone block bioactive glass in rabbits.Methods Establish the femoral condyle defect model with New Zealand white rabbit.Implant in the defect with macro-pore bone block bioglass,β-TCP and NOVABONE(R) respectively.According to the different materials implanted in the defect,three groups were divided as macro-pore bone block bioglass group,β-TCP group and NOVABONE group.After the surgery,X-ray examination was performed to confirm the location and fixation of the materials and to observe the femoral condyle fracture.The specimens were harvested at 4,12 and 24 weeks after the surgery respectively.MicroCT was performed to assess the new bone formation and degradation of the materials.Tetracycline-calcein double labeling was used to detect the mineral apposition rate of new bone.Van Gieson staining was used to assess the new bone formation percentage.Biomechanical markers including the compress strength and elasticity modulus were also measured.Results X-ray examination showed that each femoral defect was filled fully with materials and the materials were all in proper position.As indicated by MicroCT results,at 24 weeks,the bone regeneration volume fraction of each group was 37.48％ ±0.70％,25.29％± 1.45％,27.03％±1.25％ respectively and the difference between macro-pore bone block group and β-TCP group or NOVABONE group was statistically significant.The residual material volume fraction of each group was 34.67％±3.52％,55.66％±2.05％,7.52％± 1.15％ respectively and the difference between macro-pore bone block group and β-TCP group or NOVABONE group was statistically significant.The results of tetracycline-calcein double labeling showed that the mineral apposition rate in macro-pore bone block bioglass group,β-TCP group and NOVABONE group at 4 weeks was (1.577±0.045) um/d,(2.064±0.068)um/d,(1.19±0.09)um/d respectively and the difference between macro-pore bone block bioglass group and β-TCP group was statistically significant.As shown by the results of Van Gieson staining,the new bone area percentage of macro-pore bone block bioglass group,β-TCP group and NOVABONE group was 5.43％± 1.25％,2.77％±0.85％,6.51％± 1.21％ at 4 weeks,8.48％±0.84％,2.94％±0.65％,11.42％±2.66％ at 12 weeks,23.55％± 1.13％,12.92％±0.45％,19.53％±0.91％ at 24 weeks.The difference between macro-pore bone block bioglass group and β-TCP group or NOVABONE group at 24 weeks was statistically significant.By biomechanical test,the compress strength of specimens in macro-pore bone block bioglass group and β-TCP group increased as time prolonged,with no statistically significant between the two groups.The elasticity modulus of specimens in macro-pore bone block bioglass group and NOVABONE group was stable after surgery,closer to the rabbit bone,while elasticity modulus of the β-TCP group increased a lot,unsuit to the rabbit bone.Conclusion Macro-pore bone block bioglass presented good biological activity,biocompatibility and suitable biomechanical properties.This research loaded foundation for the application in weight-bearing sites of this new material.

Objective To explore the effect of therapeutic hypothermia on the activation of inflammasome in the lung tissue of rats with hemorrhagic shock (HS).Methods Twenty-four male Sprague-Dawley rats were randomly (random number) divided into three groups:sham group,normothermia resuscitation (NR) group and hypothermia resuscitation (HR) group.Rats of each group were subjected to pressure-controlled (MAP 40 mmHg) HS for 1 h,then the NR group and the HR group were resuscitated with lactated Ringer and MAP was maintained at 90 mmHg for 1 h.Four hours later,the rats in each group were sacrificed by exsanguination.Hematoxylin eosin staining was used to observe the injury of lung tissue.The desiccation method was used to detect the edema of lung tissue.RT-PCR and western blot were employed to evaluate the mRNA and protein expression of NLRP-3,IL-1β,caspase-1.Analysis of variance was used for comparison among groups,and SNK-q test was used for comparison between two groups.Results (1) The injury of lung tissue in HR group was significantly milder than that in NR group;(2) Wet/dry (W/D) weight ratio of lung in HR group decrease compared with NR group [HR group 5.85 ± 0.102;NR group 6.471 ± 0.165 8 (t =3.14,P ＜ 0.05)];(3) NLRP-3 and of Caspase-1 protein expression in the HR group were lower than those in NR group.(4) The NLRP-3 mRNA expression in HR group was lower compared with NR group [(HR group 1.027 ± 0.143;NR group 1.3487 ± 0.163 (t =4.36,P ＜ 0.05)] and IL-1 mRNA expression in HR group was lower compared with NR group [HR group 162.3 ± 0.125;NR group 2.388 ± 0.229 (t =7.72,P ＜ 0.05)].Conclusions Therapeutic hypothermia attenuated ALI induced by HS in rats by modulation of signal way of inflammasome.

Objective To observe and identify the impact of a type of macro?pore bone block bioactive glass on osteo?blast in vitro. Methods Extract fluid of new bioactive glass was prepared withα?MEM culture medium as the bioactive medium group. And the concentrations of different ions were detected with Inductively Coupled Plasma?Atomic Emission Spectrometry in bioactive medium group andα?MEM medium group. MC3T3?E1 cells cultured in bioactive medium group were considered as ex?perimental group and cells cultured inα?MEM medium as control group. Giemsa and immunofluorescence staining was performed to detect the cell numbers, the karyoplasmic ratio and the average fluorescence intensity per cell. Cell proliferation and viability in different groups were detected by cell cycle analysis, MTT assay and BrdU assay, respectively. Total RNAs of cells in different groups were extracted and the expressions of ALP, OCN and collagenⅠwere measured by quantitative real time PCR. ALP stain?ing and alizarin red staining were performed to assess the differentiation and mineralization of MC3TC?E1 cells in different groups. Results The concentrations of Si and F were 40.02 ± 0.67 mg/L and 0.02 ± 0.001 mg/L in bioactive medium group, higher than 2.02±0.01 mg/L and 0.00 mg/L inα?MEM solution, and the concentration of Ca was lower than that inα?MEM solution. The con?centration of P and Na had no difference. In Giemsa staining, the cell number in 400 times field under a microscope was 106.0 ± 6.025 in bioactive medium group and 40.20 ± 3.639 inα?MEM medium group. In the immunofluorescence of vinculin, the karyo?plasmic ratio and the expression of vinculin were higher in bioactive medium group (40.85±5.720, 0.050 88±0.021 78) than inα? MEM medium group (21.93 ± 4.137, 0.023 60 ± 0.003 18). In cell cycle analysis, the proportion of cells retained in S and G2/M phase in the bioactive medium group was more than that in theα?MEM medium group after 72 hours of cell culture. In the BrdU and MTT assay, MC3T3?E1 cells in bioactive medium group both showed a higher proliferation rate with statistical significance. In MC3T3?E1 cells cultured with the bioactive medium, the expressions of osteogenesis?related genes were higher than those cultured with ordinaryα?MEM solution;in the ALP staining and alizarin red staining, the expression of ALP and the mineralization rate were higher in bioactive medium group (1.328%±0.015 36%, 2.953%±0.536 3%) than inα?MEM medium group (0.979%±0.030 59%, 1.000%±0.208 1%). Conclusion The bioactive medium promotes cell proliferation and osteoblastic differentiation of MC3T3?E1 cells, and has much more Si ions, which indicates that macro?pore bone block bioactive glass can promote cell proliferation and dif?ferentiation and has promising bioactivity.

Objective To devolope a method for extracting DNA from dried blood spots (DBS)and optimizing the operating procedure,which could be applied to clinical gene diagnosis of thalassemia.And the cross contamination of DBS punching and the storage stability of DBS were studied.Methods A total of 1 50 blood specimens were collected,and DBS were prepared.Circles (3 mm in diameter)were punched in the DBS,and eluted with lysis buffer.The eluting method and operating procedure were opti-mized.Genomic DNA extracted from the elution solution by magnetic beads,and were performed thalassemia gene test.Finally jud-ging whether the results of DBS and whole blood were consistent.Two methods of thalassemia gene test were used in DBS and the compatibility of DBS processing method was verified.Judging whether there was cross contamination of DBS punching by the thalassemia gene test results of blank hole which were punched in the blank filter paper between thalassemia positive DBS.The DBS storage stability in thalassemia gene test was verified by detecting the DBS which were dry stored at room temperature for 6 and 9 months.Results 5 circles (3 mm in diameter)DBS were vibrating eluted at 55 ℃ for 1 hour,the DNA concentration extracted from the elution solution was 10-20 ng/μL,which was dissolved in 50 μL solution,and the DNA quality was good.The thalassemia gene test results of DBS and whole blood were the same,and the DBS results of two thalassemia gene test methods were the same too. The cross contamination of DBS punching was not detected in thalassemia gene test.The DBS which were dry stored at room tem-perature for 6 and 9 months could be stably performed thalassemia gene test.Conclusion DBS could be used to perform thalassemia gene test,which is accurate,convenient and stable.It is an ideal way for specimen referral of thalassemia gene test.

Objective:To explore the effects of operation timing on postoperative complications and operative duration in children undergoing non-emergency operation for femoral neck fracture.Methods:Fifty-nine children and adolescents with femoral neck fracture were retrospectively analyzed who had been admitted to Department of Pediatric Orthopedics, Zhengzhou Orthopedic Hospital Affiliated to Henan University from February, 2015 to September, 2019. They were 39 boys and 20 girls with a mean age of 11.7 (7.1, 13.7) years. By Delbet fracture classification, 7 cases were type Ⅰ, 27 cases type Ⅱ, 20 cases type Ⅲ, and 5 cases type Ⅳ. The patients were divided into 4 groups by the time from injury to operation (TFITO): 17 cases were assigned into group A with TFITO from 24 to 48 hours, 14 cases into group B with TFITO from 49 to 72 hours, 12 cases into group C with TFITO from 73 to 96 hours, and 16 cases into group D with TFITO>96 hours. The effects of TFITO on postoperative complications and operative duration were analyzed.Results:There were no significant differences between the 4 groups in their preoperative general data, showing they were comparable ( P>0.05). This cohort was followed up for 12 to 62 months (average, 20 months). The operation time was not included for this study in 6 cases whose associated injury had to be treated simultaneously. The median operation time for the other 53 patients was 80 (70, 105) min. The correlation coefficient between TFITO and operation time was 0.098 ( P=0.484). Postoperative complications occurred in 37.3% of the patients (22/59), including 14 cases of avascular necrosis of femoral head. For groups A, B, C and D, the incidences of complications were 47.1% (8/17), 50.0% (7/14), 25.0% (3/12) and 25.0% (4/16) while the incidences of avascular necrosis of femoral head 23.5% (4/17), 31.3% (5/16), 16.7% (2/12) and 18.8% (3/16), showing insignificant differences between the 4 groups in all the comparisons ( P>0.05). Conclusion:The time from injury to operation may not increase operative duration or postoperative complications such as avascular necrosis of femoral head in children undergoing non-emergency operation for femoral neck fracture.

To explore the clinical effect of microsurgery in the treatment of the tumor which was diagnosed with the plexiform neurofibroma (PN) of the forearm and palm. Methods From January, 2014 to June, 2017, 6 cases of the PN in the forearm and palm were removed by microsurgery such as neurovascular transplantation, separation and anastomosis under microscope, etc. There were 4 males and 2 females, with an average age of 9.2 (range, 2-18 )years. There was 1 case with PN of the median nerve, ulnar nerve and their branches in the right fore-arm and palm, 2 cases with PN of the median nerve and its branches in the right forearm and palm, 2 cases with PN of the median nerve and its branches in the left forearm and palm, and 1 case with PN of the ulnar nerve and its branches in the left forearm and palm.The postoperative function and feeling of the patients were evaluated by outpa-tient followed-up. Results The pathological results of 6 patients all showed PN, and their incisions healed primari-ly.The patients were followed-up for 6 to 36 months, with an average of 18 months. No obvious scar formation was observed in all incisions. Among them, PN of the palmar of the youngest patient recurred after the operation, and it was resected in a second operation.The remaining 5 patients had no recurrence during follow-up.The 2 point resolu-tion of each fingertip of the affected limb of the patients who had median and ulnar PN was 2-5 mm, with an average of 3.30 mm; the 2 point resolution of the thumb, indicator, middle and ring fingers of the affected limbs of the patients who had median PN was 2-5 mm, with an average of 2.95 mm; the 2 point resolution of the ring and little fingers of the affected limbs of the patients who had ulnar PN was 3-4 mm, with an average of 3.50 mm.According to the related evalu-ation criteria made by the American Orthopedic Foot and Ankle Surgery Society (AOFAS), the results of the forearm and hand functions were excellent in 5 cases, good in 1 case. Conclusion The application of microsurgical techniques in the treatment of PN in the forearm and palm can be effective separation of tumor and nerve fibers, effectively protect the branches of the median nerve and ulnar nerve and their blood circulation, prevent recurrence and reduce nerve damage after operation.