BACKGROUND:Studies have shown that bone morphogenetic proteins play a key role in skeletal development. Platelet-rich plasma alone in animal or clinical trials cannot significantly promote bone graft healing.
OBJECTIVE:To investigate the osteogenesis effect of bone morphogenetic protein-4 compounded with platelet-rich plasma the bone defect area.
METHODTwenty-four New Zealand white rabbits were selected to establish maxil ary bone defect models, and then were randomly divided into four groups, six rats in each group. Group A was blank control group;Group B wasβ-tricalcium phosphate (0.1 g)+Bio-gide group;group C wasβ-tricalcium phosphate (0.1 g)+Bio-gide+platelet-rich plasma (1 mL) group;and group D wasβ-tricalcium phosphate (0.1 g)+Bio-gide+platelet-rich plasma (1 mL)+bone morphogenetic protein-4 (5μg). Gross observation, histological observation and imaging analysis were performed for analysis of new bone formation at weeks 4, 8, 12 after modeling.
RESULTS AND CONCLUSION:After 4 weeks, group D had more new bone and vessels formed in the bone defect area than the group B (P<0.01);however, the amount of new bone was higher in the group B than the group A (P<0.01). After 4, 8, 12 weeks, the amount of new bone in the bone defect area was higher in the group D than the groups B and C (P<0.01), and lowest in the group A (P<0.01). In theβ-tricalcium phosphate scaffold, platelet-rich plasma combined with bone morphogenetic protein can significantly promote the healing of bone defects.

Objective To evaluate the effect of alendronate on osteoprotegerin (OPG) and receptor of activator of nuclear factor κB-ligand (RANKL) expression in human marrow stroma cells (hMSCs) in vitro. Methods hMSCs were isolated from haman marrow, cultured in vitro, and randomly divided into two groups: alendronate group, hMSCs culture fluid containing 1×10-7 mol/ L alendronate; control group, no special treatment but culturing hMSCs in DMEM. Two weeks after treatment, the expressions of OPG and RANKL were evaluated by RT-PCR and Western blot.Results hMSCs became uniform spindle-shaped fibroblasts. As cells proliferated, they formed colonies and showed whirlpool arrangement. After one week's treatment, hMSCs in alendronate group had reduced processes and gradually showed disc shape, which did not happen in control group but kept fibroblast shape and just increased in density. In RT-PCR, the ratio of OPG/RANKL in alendronate group and control group was 8.77±1.16 and 4.58±1.27, respec-tively. In Western blot, the ratio of OPG/RANKL in alendronate group and control group was 2.58±0.47 and 1.52±0.32, respectively. The ratio of OPG/RANKL was higher in alendronate group than in control group (P<0.01).Conclusion Alendronatc enhances OPG expression and inhibits RANKL expression of hMSCs in vitro.

Objective To compare the clinical effects of treating cervical prolapse of intervertebral disc by using Solis cage, titanium cage and autogenous iliac crest graft (AICG) combined with titanium plate. Methods 64 cases of cervical prolapse of intervertebral disc were analyzed retrospectively. All the patients were followed up for about one year. Group A is composed of 20 patients (30 intervertebral spaces) treated with microdiscectomy and Solis cage fusion;group B is composed of 21 patients(22 intervertebral spaces) treated with titanium cage and group C is involved of 23patients(28 intervertebral spaces) which were dealt with AICG combined with titanium plate. The differences in the to-tal X ray exposure time, time working on iliac bone, incidence of complications, fusion rate, incidence of JOA score recovery rate more than 50 % and rehabilitation time of the three groups are analyzed. Results All indexes from group A and B were more satisfactory than those from group C (P＜0. 05). The statistic analysis results of the incidence of complications of cervical part, rehabilitation time in group A are 5.0± 1.8, 5.1 ± 1.2; and that is 14.3±2.6, 7.5 ± 1.6and 26.1±6.2, 8.6±2.3 in group B and group C respectively. There is significant difference between group A and group B on the incidence of complications of cervical part and rehabilitation time (P＜0.05). Results of the incidence of JOA score recovery rate ＞ 50 % in group A(95.8 ± 3.2) was more satisfactory than that in group B(93.6 ± 2.9),while there was no statistical difference between group A and group B (P＞0.05). Conclusion The Solis cage is better in treating the cervical prolapse of intervertebral disc than other ways. Also, it makes the operation easier, rehabilitation time shorter, and it causes fewer complications.

To explore the effects of different doses of puerarin on proliferation of cultured vascular endothelial cells in vitro.

OBJECTIVE: To observe the promoting effect of arnebia root oils on expression of basic fibroblast growth factor (bFGF) in skin wound of rabbits and the histomorphological changes in the wound surface, and to discuss its mechanism. METHODS: Bilateral round skin wounds were made on the back of 15 rabbits. The three wounds on one side of the back of each rabbit were treated with arnebia root oils, while the three wounds on the other side were treated with vaseline in order to promote the wound healing. The histomorphology and ultrastructure under electron microscopy of the wounds, and the rate of wound healing were examined at different time. Western blotting assay was used to detect the expression of bFGF in the wound surface. RESULTS: The healing rate of the arnebia root oils-treated wounds was evidently higher than that of the vaseline-treated wounds (P<0.05). The quantities of fibroblast, collagen and capillary in the arnebia root oils-treated wounds were much more than those in the vaseline-treated wounds, and the expression of endogenous bFGF in the arnebia root oils-treated wounds was enhanced obviously as compared with that in the vaseline-treated wounds in different period of wound healing. There existed a parallel correlation between the expression level of bFGF and the rate of wound healing. CONCLUSION: The promoting effect of arnebia root oils on wound healing may be related to increasing the expression level of basic fibroblast growth factor in the skin wound.

Objective To investigate T-bet mRNA expression on peripheral blood mononuclear cells (PBMCs) and its relations with allergen specific IgE (SIgE), eosinophile cationic protein (ECP) levels, and allergic symptom in patients with allergic rhinitis (AR). Methods The allergen, SIgE, and ECP in serum of patients with AR were detected by Unicap CAP system. Blood samples were taken from 15 healthy controls and 35 house dust mite allergic patients. PBMC was isolated by density gradient centrifugation and one part of them was cultured with mite allergen at a concentration of 50 μg/mL. PBMC was subjected to analysis of T-bet mRNA expression using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Results The ratio of T-bet to β-actin mRNA levels was 0.418 ± 0. 101 in patients of AR and 0.706 ± 0.091 in healthy controls and the difference was significant (P ＜ 0.01). The expression intensity of T-bet mRNA was not related to varying severity of allergic symptom and ECP levels ( r = - 0.227, - 0.033, P ＞ 0.05). However, there was an inverse correlation between expression intensity of T-bet mRNA and SIgE concentration (r = -0.375, P ＜ 0.05). There was a positive correlation between SIgE and allergic symptom scores ( r = 0.426, P ＜ 0.05). After that PBMC was stimulated by mite allergen, the expression intensity of T-bet mRNA, ECP, and SIgE changed very little ( P ＞ 0.05). Conclusion Down-regulated expression of T-bet mRNA in mite-AR patients is not related to serum ECP and symptom scores but one of important links in the mechanism of imbalance of Th1/Th2 in the occurrence of AR. Specific allergen has no effect on T-bet mRNA, ECP, and SIgE of children and adults with AR in vitro. The level of SIgE objectively and directly indicates the severity of allergic symptom, but T-bet did not. T-bet may be one of indirect factors which affect the level of IgE.

Objective To investigate the serum levels of β-amyloid precursor protein (β-APP) and hypersensitive-cross-reactive protein(hs-CRP) after acute spinal cord injury(SCI) in rats at different time points and study their roles in secondary spinal cord injury. Methods Ninety healthy adult female SD rats were randomly assigned into 3 even groups.Improved Allen method was used to create models of complete SCI (group A) and models of incomplete SCI (group B).In group C only laminectomy was performed without SCI.The serum was collected at 12 hours,24 hours,3 days,7 days,and 14 days after operation through femoral vein to detect levels of β-APP and hs-CRP by ELISA. ResultsThe serum level of β-APP reached the peak value 24 hours after acute SCI,and the serum level of hs-CRP reached the peak 3 days after acute SCI.There were significant differences between groups A and C in β-APP and hs-CRP levels at all time points( P ＜ 0.05).There were also significant differences between groups B and C in β-APP and hs-CRP levels at all time points (P ＜0.05) except at 14 days (P＞ 0.05). Conclusions The serum levels of β-APP and hs-CRP are positively associated with the severity of traumatic SCI.β-APP may be a more sensitive indicator than hs-CRP in a certain time.The dynamic changes of serum β-APP and hs-CRP suggest involvement of the 2 proteins in the secondary spinal cord injury.

Objective To observe the effect of the in vitro isolation, culture and identification of rabbit bone marrow-derived mesenchymal stem cells (BMSCs) and explore the differentiation potentials of BMSCs. Methods BMSCs were isolated and purified by density gradient centrifugation combined with attachment culture method. The biological characteristics of BMSCs were observed under an inverted microscope. The BMSCs were identified with HE staining and CD34/CD44 immunohistochemical staining. The biological activity of BMSCs was detected by osteogenic induction and steatogenic induction. Results BMSCs were isolated successfully in vitro and had good homogenicity. Immunohistochemical staining of CD34/CD44 showed strong positive expression of CD44 and negative expression of CD34. Fortis osteogenic ability and positive staining of ALP could be detected by osteogenic induction, and fortis steatogenic ability and positive staining of axungia could be detected by steatogenic induction. Conclusion BMSCs have the capability of multi-directional differentiation in vitro.

Objective: To explore the association of the single nucleotide polymorphisms (SNPs) in N-formylpeptide receptor (FPR) gene with the susceptibility of aggressive periodontitis (AgP). Methods: A total of 94 AgP patients and 73 healthy controls were entered into the study. Peripheral blood sample was obtained from each subject by venepuncture. Genomic DNA was isolated from each sample.The target fragment of FPR gene was amplified by PCR. The SNPs in FPR gene were detected by denature high performance liquid chromatography ( DHPLC) combined with DNA sequencing. Results: There were two non-synonymous SNPs in the 370 bp FPR gene fragment;289C/A and 301G/C. The 289C/Awas a novel SNP. No variation in nucleotides 329 and 378 was detected. There were no statistically significant differences in distributions of the genotypes and alleles for FPR289 and FPR301 between AgP patients and healthy controls. Using multivariate logistic regression (adjusted for age and gender) , it was showed that the adjusted Ors of AgP for the C~+ genotype and allele C of FPR301 combined with smoking were 5.74 and 5.20 respectively. Conclusion: The presence of the C~+ genotype/allele C of FPR301 together with smoking conferred a higher risk for AgP. The result suggests that the SNPs in FPR gene may not be associated with the susceptibility of AgP in Chinese.

ts the shear stress, thus, facilitates the union of fracture and the restoration of function.