1.Effect of moderate treadmill exercise together with modified hydroxyapatite chitosan composite hydrogel implantation on repair of full-thickness defects of articular cartilage in rats
Dongyang FENG ; Lei WANG ; Yu CHAI ; Shenyu YANG ; Yi JIANG ; Peiran ZHAO ; Bin YU
Chinese Journal of Orthopaedic Trauma 2016;18(6):518-525
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.
2.Expression,Purification,Preparation of Polyclonal Antibody and Subcelluar Localization of the NS2 Protein of Periplaneta fuliginosa Densovirus
Bo YANG ; Peiran YU ; Dawei CAI ; Xiaomin DONG ; Zhigang LIU ; Zheng HU ; Jiamin ZHANG ; Yuanyang HU
Microbiology 2008;0(09):-
NS2 is a nonstructural protein of Periplaneta fuliginosa densovirus(PfDNV) with a molecular mass of 30 kD,whose function is not yet clearly understood. In order to study the expression,subcellular distribution and the function of NS2 protein,the coding region of NS2 was amplified from the hindgut tissue of cockroaches infected with PfDNV by RT-PCR and then the recombinant prokaryotic expression vector pET28a-NS2 was constructed. The recombinant plasmid was transformed into E. coli BL21(DE3) to express the 6?His fusion protein in the bacteria. After purification,the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody. The specificity of the anti-NS2 antibody was successfullyproved by western blotting on the eukaryotic expressed products of NS2 protein.Meanwhile,the full sequence of ns2 gene was also cloned into the eukaryotic expression vector pAC. The recombinant plasmid pAC-NS2 was then transfected into Schneider line 2(S2) cells to express NS2 protein in the insect cells. The subcellular localization of NS2 in the insect cells was then investigated by indirect immunofluorescence technique using the anti-NS2 polyclonal antiserum. The confocal laser scanning microscope observation showed that NS2 protein was located primarily in the cytoplasm with some punctate nuclear staining.
3.Pathogenic gene mutation in a Han Chinese family with hereditary vitreous amyloidosis identified by Sanger sequencing
Peiran ZHU ; Qiuyue WU ; Maomao YU ; Mingchao ZHANG ; Mengxia NI ; Shuaimei LIU ; Weijun JING ; Jing ZHANG ; Weiwei LI ; Jin CAO ; Yi LI ; Chunyan XUE ; Xinyi XIA
Chinese Journal of Clinical Laboratory Science 2017;35(3):181-183
Objective Our purpose was to investigate the pathogenic gene mutation of a Han Chinese family with vitreous amyloidosis.Methods The 9 individuals(proband,1 affected member and 7 unaffected members) of the family were selected and their DNA was extracted from peripheral blood.The 4 exons of transthyretin(TTR) gene were amplified by polymerase chain reaction(PCR) technique.The amplified products of TTR gene were sequencing by Sanger technique.We also selected 100 unrelated healthy individual as the control group.Results By DNA sequencing,a heterozygous mutation was found in 4 of the 9 subjects from the family.The transition of adenine to cytosine(AAG > ACG) was detectable in exon 2 of TTR,which changed the amino acid composition at codon 35 (Lys35Thr).This mutation did not presented in control group.Conclusion The heterozygosis mutation of TTR gene Lys35Thr should be a pathogenic mutation for the family with vitreous amyloidosis.
4.Effect of ozonated water on physical and chemical properties of vacuum sealing drainage material.
Nan JIANG ; Yunfei MA ; Qingrong LIN ; Anfu CHEN ; Peiran ZHAO ; Guoxin NI ; Bin YU
Journal of Southern Medical University 2013;33(2):290-295
OBJECTIVETo investigate the influence of ozonated water on physical and chemical properties of vacuum sealing drainage (VSD) materials.
METHODSVSD 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.
RESULTSzonated 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.
CONCLUSIONExcept 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.
Biomedical and Dental Materials ; chemistry ; Drainage ; instrumentation ; methods ; Ozone ; Vacuum ; Water ; chemistry
5.Effects of exercise of different intensity on early repair of full-thickness articular cartilage defects and expressions of MMP-3 and TIMP-1 in rats.
Fu DONG ; Jinqi SONG ; Jiwei LUO ; Peiran ZHAO ; Guoxin NI ; Bin YU
Journal of Southern Medical University 2014;34(1):103-108
OBJECTIVETo 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.
METHODSTwenty-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.
RESULTSIn 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.
CONCLUSIONBoth 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.
Animals ; Cartilage, Articular ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 3 ; metabolism ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Wound Healing
6.Effects of exercise of different intensity on early repair of full-thickness articular cartilage defects and expressions of MMP-3 and TIMP-1 in rats
Fu DONG ; Jinqi SONG ; Jiwei LUO ; Peiran ZHAO ; Guoxin NI ; Bin YU
Journal of Southern Medical University 2014;(1):103-108
Objective 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. Methods 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 trochleas 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. Results 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. Conclusion 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.
7.Effects of exercise of different intensity on early repair of full-thickness articular cartilage defects and expressions of MMP-3 and TIMP-1 in rats
Fu DONG ; Jinqi SONG ; Jiwei LUO ; Peiran ZHAO ; Guoxin NI ; Bin YU
Journal of Southern Medical University 2014;(1):103-108
Objective 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. Methods 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 trochleas 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. Results 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. Conclusion 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.
8."Frame and buttress" fixation for the treatment of acetabular quadrilateral fractures
Yizhou WAN ; Kaifang CHEN ; Yan MA ; Keda YU ; Peiran XUE ; Dong YAN ; Xiaodong GUO
Chinese Journal of Orthopaedics 2023;43(14):933-940
Objective:To investigate the efficacy and safety of the "frame and buttress" fixation in the treatment of acetabular quadrilateral fractures.Methods:A total of 26 patients with acetabular quadrilateral fractures treated with the "Frame and buttress" fixation strategy (reconstructing frame and providing buttress for quadrilateral surface) from January 2017 to January 2021 were retrospectively analyzed in the Department of Orthopaedics, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, including 17 males and 9 females, aged 47.7±16.5 years, (range, 25-71 years). There were 13 cases from car accidents, 8 cases from falls, and 5 cases from heavy objects. Quadrilateral fractures were classified according to whether the anterior column (A) and posterior column (P) of the acetabulum were fractured and whether the quadrilateral plate (Q) was partially (1) or completely (2) separated from the two columns. Among these cases, there were 5 cases of type AQ1, 1 case of type AQ2, 1 case of type PQ2, 14 cases of type APQ1 and 5 cases of type APQ2. The "frame and buttress" fixation strategy was used to perform open reduction and internal fixation. The Matta scale was used to assess the quality of resurfacing based on pelvic radiographs, and the Matta modified Merle d'Aubigne and Postel scores were referenced to evaluate hip function.Results:All the 26 patients successfully completed the operation, the operation time was 88.1±57.8 min (range, 90-320 min), the intraoperative bleeding was 753.8±309.5 ml (range, 400-1 400 ml). There were 14 cases of anatomical reduction, 8 cases of satisfactory reduction and 4 cases of unsatisfactory reduction with an excellent rate of 85% (22/26), the excellence rate was 85% (22/26). All patients were followed up for 16.3±4.3 months (range, 12-24 months). The hip function score after operation was higher than that before operation, and the difference was statistically significant ( F=19.80, P<0.001). The hip function score was 13.1±2.2 at 3 months postoperatively, increased to 15.9±2.3 at 6 months postoperatively, and increased to 16.6±1.9 at the last follow-up, with a statistically significant difference ( P<0.001). After operation, 5 patients showed symptoms of lateral femoral cutaneous nerve injury such as hypoesthesia and numbness in the anterolateral thigh. During the follow-up, there was no ectopic ossification, traumatic arthritis, necrosis of the femoral head and central dislocation of the femoral head. Conclusion:The "frame and buttress" fixation strategy based on quadrilateral fracture typing for acetabular quadrilateral fracture open reduction internal fixation provides strong fixation, satisfactory reduction quality, and facilitates joint function recovery after operation.
9.HybridSucc:A Hybrid-learning Architecture for General and Species-specific Succinylation Site Prediction
Ning WANSHAN ; Xu HAODONG ; Jiang PEIRAN ; Cheng HAN ; Deng WANKUN ; Guo YAPING ; Xue YU
Genomics, Proteomics & Bioinformatics 2020;18(2):194-207
As an important protein acylation modification, lysine succinylation (Ksucc) is involved in diverse biological processes, and participates in human tumorigenesis. Here, we collected 26,243 non-redundant known Ksucc sites from 13 species as the benchmark data set, combined 10 types of informative features, and implemented a hybrid-learning architecture by integrating deep-learning and conventional machine-learning algorithms into a single framework. We constructed a new tool named HybridSucc, which achieved area under curve (AUC) values of 0.885 and 0.952 for general and human-specific prediction of Ksucc sites, respectively. In comparison, the accuracy of Hybrid-Succ was 17.84%–50.62%better than that of other existing tools. Using HybridSucc, we conducted a proteome-wide prediction and prioritized 370 cancer mutations that change Ksucc states of 218 important proteins, including PKM2, SHMT2, and IDH2. We not only developed a high-profile tool for predicting Ksucc sites, but also generated useful candidates for further experimental con-sideration. The online service of HybridSucc can be freely accessed for academic research at http://hybridsucc.biocuckoo.org/.