ABSTRACT
OBJECTIVE@#To explore the clinical characteristics of congenital neutropenia caused by ELANE gene mutations.@*METHODS@#Clinical manifestations, absolute blood neutrophil count, high-throughput exome sequencing for mutation screening, suspected locus Sanger sequencing verification, processes of diagnosis and treatment of two patients with congenital neutropenia caused by ELANE gene mutation were retrospectively analyzed.@*RESULTS@#High-throughput sequencing has found that proband 1 has carried a heterozygous c.170C>T (p.Ala57Val) missense mutation in exon 2 of the ELANE gene, which was known to be pathological, and a heterozygous c.251T>G (p.Leu84Arg) mutation in exon 3 of proband 2, which was unreported previously. Sanger sequencing confirmed that neither mutation was inherited from their parents.@*CONCLUSION@#ELANE mutation is an important cause for congenital neutropenia. Detection of new pathogenic variants has enriched the mutation spectrum of the ELANE gene.
ABSTRACT
Objective To investigate the effect of moderate treadmill exercise together with modified hydroxyapatite chitosan composite hydrogel (CS/HA-g-CS) implantation on repair of full-thickness defects of articular cartilage in rats.Methods Full-thickness cartilage defects were drilled in the patellar groove of bilateral femoral condyles in a total of 24 male SD rats before they were randomly assigned into 4 groups.The control group (BC group) was subjected to no exercise or CS/HA-g-CS implantation;the chitosan group (CHI group) to CS/HA-g-CS implantation without exercise;the moderate treadmill exercise group (MIR group) to exercise 4 weeks after modeling without CS/HA-g-CS implantation;the CHI + MIR group to moderate treadmill exercise plus CS/HA-g-CS implantation 4 weeks after modeling.Half of the animals were sacrificed at week 8 and half at week 16 after operation.Femoral condyles were harvested for gross observation and histochemical measurement by O' Driscoll scoring system.mRNA expressions of glycosaminoglycan,collagen type Ⅱ and BMP-2 were detected by RT-PCR.Results Gross observation revealed:at 8 weeks after modeling,the CHI + MIR group was significantly better than the other 3 groups,with the BC group in the poorest (P ＜ 0.05);at 16 weeks after modeling,the BC group was significantly poorer than the other 3 groups (P ＜0.05) among which there were no significant differences (P ＞ 0.05).O 'Driscoll scoring revealed:at both 8 and 16 weeks after modeling,the CHI + MIR group was significantly better than the other 2 groups and the BC group significantly poorer than the other 3 groups (P ＜ 0.05) but there was no significant difference between the MIR and CHI + MIR groups(P ＞ 0.05).The expressions of collagen type Ⅱ,glycosaminoglycan and BMP-2 were significantly higher in the CHI + MIR group than in the other 3 groups (P ＜ 0.05) and significantly lower in the BC group than in the other 3 groups (P ＜ 0.05).Conclusions Moderate treadmill exercise together with CS/HA-g-CS implantation has significant positive effects on repair of full-thickness defects of articular cartilage in rats than merely moderate treadmill exercise or CS/HA-g-CS implantation alone.The defective cartilage repaired by moderate treadmill exercise together with CS/HA-g-CS implantation contains more collagen type Ⅱ and glycosaminoglycan and shows morphology of nearly normal cartilage.
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effects of treadmill running exercise of different intensity on early repair of full-thickness defects on the patellofemoral articular surface and the changes in the expressions of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in SD rats.</p><p><b>METHODS</b>Twenty-four male SD rats with full-thickness defects on the patellofemoral articular surface were randomly assigned into sedentary control (SED) group and low-, moderate- and high-intensity running groups (LIR, MIR, and HIR groups, respectively). The running groups were trained on treadmill for 6 consecutive weeks. Blood samples were collected to detect serum MMP-3 and TIMP-1 levels using ELISA before and after the experiment, and the femoral trochlea were collected to assess tissue repair by gross appearance scoring and O Driscoll histological scoring with Safranine O-Fast Green staining and Toluidine blue staining.</p><p><b>RESULTS</b>In rats in SED group, the defect was filled with hyaline articular cartilage-like tissues, as compared to fibrous tissues in LIR and MIR groups and subchondral bone damage in HIR group. The SED group scored the highest and HIR group the lowest among the 4 groups in gross appearance scoring and O Driscoll histological scoring. No significant differences were found in MMP-3 or TIMP-1 levels among the groups before training (P>0.05), but after 6 weeks of training, serum MMP-3 and TIMP-1 levels differed significantly among the 4 groups (P<0.05), and all the 3 running groups had a significantly higher MMP-3 level than the control group (P<0.05). After the 6-week training, TIMP-1/MMP-3 ratio was significantly higher in SED group than in the 3 running groups, and was the lowest in HIR group.</p><p><b>CONCLUSION</b>Both low- and moderate-intensity exercise failed to promote resurfacing of full-thickness cartilage defects on the patellofemoral articular surface in rats, and high-intensity exercise even induces subchondral bone damage. The expression of MMP-3 and TIMP-1 is related to exercise, and the TIMP-1/MMP-3 ratio reflects the extent of tissue repair.</p>
Subject(s)
Animals , Male , Rats , Cartilage, Articular , Metabolism , Pathology , Matrix Metalloproteinase 1 , Metabolism , Matrix Metalloproteinase 3 , Metabolism , Physical Conditioning, Animal , Rats, Sprague-Dawley , Tissue Inhibitor of Metalloproteinase-1 , Metabolism , Wound HealingABSTRACT
<p><b>OBJECTIVE</b>To investigate the influence of ozonated water on physical and chemical properties of vacuum sealing drainage (VSD) materials.</p><p><b>METHODS</b>VSD materials (foam and sealing membrane) were immersed in 10 µg/ml ozonated water for 1 h twice daily for 8 days. The foam appearance and microscopic structure of the materials were observed, and tensile tests and Raman spectrum scan were performed assess the effect of ozonated water. Simulated VSD devices were prepared and tested for leakproofness under negative pressure after ozonated water treatment.</p><p><b>RESULTS</b>zonated water treatment for 8 days caused no obvious abnormal changes in the foam appearance or microscopic structure of the materials. The maximum tensile load of foam before and after ozonated water treatment was 4.25∓0.73 kgf and 2.44∓0.19 kgf (P=0.000), the momentary distance when the foam torn before and after intervention was 92.54∓12.83 mm and 64.44∓4.60 mm, respectively (P=0.000). The corresponding results for VSD sealing membrane were 0.70∓0.58 kgf and 0.71∓0.08 kgf (P=0.698), and 99.30∓10.27 mm and 100.95∓18.22 mm (P=0.966), respectively. Raman spectroscopy revealed changes in only several wave intensities and no new chemical groups appeared within the scan range of 400-4000 cm(-1). The VSD device was well hermetic after treatment with ozonated water.</p><p><b>CONCLUSION</b>Except for a decreased stretch resistance property of the foam, VSD materials display no obvious changes in physical and chemical characteristics after treatment with ozonated water for 8 days.</p>
Subject(s)
Biomedical and Dental Materials , Chemistry , Drainage , Methods , Ozone , Vacuum , Water , ChemistryABSTRACT
Objective To explore the proliferation and differentiation of osteoblasts from 2 sources co-cultured with SD rat Schwann cells(SCs) . Methods Bone marrow stromal cells (BMSCs) were obtained by washing the femoral and tibial bone marrow cavities in SD rats. Osteoblast differentiation of the third passage of BMSCs was induced by incubation in osteogenic medium. Primary rat calvarial osteoblasts were obtained by digestion of the calvarial bone in one day old SD rats. The cells were cultured in DMEM supplemented with 10% fetal bovine serum(FBS) . SCs of passage 2 were obtained by digestion of sciatic nerve. The SCs were identified by S-100. The proliferation of 2 kinds of osteoblasts co-cultured with SCs was tested using 96 co-culture plate by methyl thiazdyl tetrazolium(MTF). Real-time PCR was used to test the osteoblast differentiation through co-culturing with SCs in 3 d and 7 d. The osteoblasts were implanted in the subtus chamber. The SCs were implanted in the superior chamber. Results SCs enhanced significantly the proliferation of calvarial osteoblasts at 7 time points. The expression levels of OPN mRNA, OCN mRNA, ALP mRNA, and BMP-2 mRNA of the osteoblasts were significantly lower in the experiment group than in the control group in 3 d and 7 d. SCs also enhanced significantly the proliferation of the induced osteoblasts in 5 d, 7 d and 9 d. The expression levels of OPN mRNA, OCN mRNA, ALP mRNA, and BMP-2 mRNA of the induced osteoblasts were significantly higher in the experiment group than in the control group in 3 d and 7 d, except the level of ALP mRNA in 7 d.Conclusions The BMSCs-induced osteoblasts cocultured with SCs may be used as seed cells to construct neurotized tissue engineered bone.
ABSTRACT
Objective To investigate whether tissue engineered bone with implanted vascular bun-dles can up-regulate release of vascular endothelial growth factor (VEGF) in models of femoral defect in rabbits.Methods Thirty-two rabbits were randomized into 2 even groups.In both groups, a segmental bone defect of 15 mm in length was made at the left femur before a tissue engineered bone was inserted into the defect.In the experimental group, a femoral vascular bundle was implanted into the tissue engineered bone.In the control group, there was no vascular implantation.At 2, 4, 8, and 12 weeks after implantation, samples were taken to determine new bone formation by histology and expression level of VEGF by immuno-histochemistry.Results The new bone formation was significantly higher in the experimental group at the end of 4, 8, and 12 weeks(P < 0.05) .The expression level of VEGF in the experimental group was also significantly higher than in the control group at all time points after operation, and the expression of VEGF peaked at 4 weeks.Conclusion Tissue engineered bone with vascular bundle implanted can up-regulate VEGF release in models of femoral defect in rabbits.
ABSTRACT
Objective To explore whether the respective implantation of vascular bundles and sensory nerve tracts into a tissue-engineered bone will affect the expression of CGRP (Calcitonin gene related peptide) and its receptor. Methods Fifty-four New Zealand rabbits were randomly divided into 3 even groups for implantation of sensory nerve tracts (group A),implantation of vascular bundles (group B),and a control group of simple tissue-engineered bone (group C) . Animals were sacrificed 4,8,12 weeks after implantation,respectively. Masson staining was conducted to observe the process of bone formation and re-molding. CGRP and CGRPR-1 expressions in the new bone were measured by immunohistochemistry and Real-time PCR at 4,8 and 12 weeks after implantation. Results At all time points,the CGRP and CGRPR-1 expressions in groups A and B were significantly higher than in group C (P<0.05),and those in group A were higher than in group B too (P<0.05) . Over time,the expressions of CGRP and CGRPR-1 mRNA in each group in the new bone tissue were gradually reduced after an initial increase. The neuropeptide expression at the 8th week was higher than those at the 4th and 12th weeks. The neuropeptide expression at the 4th week was the lowest. The expression of CGRP was mainly localized in the periphery of newly generated bone,periosteum and the blood vessels. The expression of CGRPR-1 was mainly localized in the periphery of osteoblasts. Conclusions Implantation of either vascular bundles or sensory nerve tracts can promote neuropeptide secretion. The vascular bundle implantation may result in higher expressions of CGRP and CGRPR-1 than sensory nerve tract implantation.