BACKGROUND:Implantation of bone morphogenetic protein (BMP) or vascular endothelial growth factor (VEGF) alone, without support vectors, is easy to be flushed away by the blood flow, and thus limits the osteogenesis and angiogenesis.
OBJECTIVE: To observe the effects of combination of calcium phosphate cement (CPC), BMP-6/VEGF in bone defect repair.
METHODS:Defect models of the bilateral medial femoral condyle were prepared in New Zealand white rabbits. Then, the medial femoral condyle was filed with CPC/BMP-6/VEGF, CPC/BMP-6, and CPC, respectively, in the left side, but nothing in the right side as control. After 8 and 16 weeks of implantation, the hard tissue slices were prepared for histological observation and scanning electron microscope observation.
RESULTS AND CONCLUSION:Al three kinds of materials showed good biocompatibility, and no obvious inflammation was found. After 8 weeks of implantation, the junction of the CPC/BMP-6/VEGF and bone tissue was almost completely covered by newly formed trabecular bone. With the development of cement degradation, abundant osteoblasts could be found in the surface of newborn trabecular bone. After 16 weeks of implantation, an ongoing cement degradation and bone formation was seen. Moreover, newly formed bone tissue increased and became thicker. The cement in the interface was separated into smal pieces and closely interconnected with the surrounding tissues, and newly formed bone showed a mesh-like ingrowth into the cement. This newly formed bone was mature and could not be distinguished from the original trabecular bone. Both the degradation and osteogenesis of CPC and CPC/BMP-6 were much slower than that of CPC/BMP-6/VEGF (P < 0.05). This study demonstrates angiogenesis and osteogenesis in vivo through the additive effects of VEGF and BMP-6. CPC/BMP-6/VEGF can be an ideal bone substitute in bone repair.

Objective To develop the clinical application of tandem mass spectrometry in earlystage diagnosis of patients with growth retardation.Methods Tandem mass spectrometry was used to quantitatively detect blood amino acids and acyl carnitine in 155 cases of children with growth retardation.The tandem mass spectrometry results,clinical symptoms,and treatment tracking results were analyzed.Results Ten patients with inborn metabolic absence of amino acids and acyl carnitine were confirmed including four cases of methylmalonic acidemia,one case of propionic acidemia,one case of medium-chain acyl-coenzyme A dehydrogenase deficiency,one case of ornithine transcarbamylase deficiency,and three cases of phenylketonuria.Conclusions Tandem mass spectrometry was helpful for some patients in etiologic diagnosis and therapeutic effect assessment of cerebral developmental retardation.

BACKGROUND: Inappropriate size of tibia prosthesis wil affect the stability and long term curative effect of knee joint. Raising the tibia prosthesis bone coverage through preoperative analysis may further reduce the incidence of loosing and sinking of tibia prosthesis. OBJECTIVE: To select the appropriate prosthesis and maximize the tibia prosthesis bone coverage rate through the three dimensional reconstruction of CT and preoperative tibia osteotomy simulation among the patients preliminarily treated with total knee arthroplasty. METHODS: Total y 76 (84 knees) patients treated with total knee arthroplasty were enrol ed and randomly divided into test group (38 cases, 41 knees) and control group (38 cases, 43 knees). Al the patients in the test group were underwent CT scan and three-dimensional reconstruction of CT before operation. Preoperative tibia osteotomy was stimulated. The most appropriate tibia prosthesis was selected by comparatively analyzing the data of tibia osteotomy, and maximizing the tibial plateau prosthesis bone coverage. Three-dimensional reconstruction of CT was absence in the control group by contrast. Tibia plateau bone coverage of patients in these two groups after replacement was evaluated. The repair effect was evaluated during the fol ow-up. RESULTS AND CONCLUSION: Total y 75 patients were fol owed up for 13 to 56 months. One patient in the test group withdrew from the study because of periprosthetic fractures. Analyzing from the fol ow-up after replacement, tibia plateau prosthesis coverage rates of test group and control group were (89.87±4.14)%, (83.15±5.21)%respectively; New York Hospital for Special Surgery scores were respectively (87.48±8.69) points and (82.37±10.14) points, the difference was statistical y significant (P < 0.05). However, there were no significant differences in the range of motion and postoperative complications between test and control groups (P > 0.05). These results suggest that choosing the most appropriate type of tibia prosthesis by applying three-dimensional reconstruction of CT and preoperative tibia osteotomy on patients treated with total knee arthroplasty can improve tibial plateau prosthesis bone coverage rate and is conductive to achieve a satisfactory repair effect.

Objective To construct MIA3 psicheck2 wild-type and mutant vectors targeting miR-374b,and provide the previous guarantee for the dual luciferase reporter assay.Methods The amplification primer was firstly designed according to mice MIA3-3'UTR sequence information,mice whole blood genomic DNA was taken as the template for PCR amplification of MIA3-3'UTR sequence,and the PCR product was cloned into psicheck2 dual luciferase reporter vector.Then,mutant primer was designed to mutate the MiR-374b seed sequence target TATTATA into AAATTAT so as to construct mutant vector.At last,the vector enzyme digestion evaluation and sequencing method was used to evaluate the constructed vectors.Results It could be seen from the analysis of agarose electrophoresis that the PCR amplification size of vector was consistent with the theoretical size.DNA sequencing evaluation showed that the MIA3-3'UTR-WT vector had been constructed successfully.The construction of mutant vector has successfully mutated the MiR-374b seed sequence target TATTATA into AAATTAT.Conclusion The successful construction of the vector will lay a foundation for the further evaluation on whether there is an actual binding site between the miR-374b and the chondrogenic differentiation-related target gene MIA3.

Aim Toinvestigatetheanalgesiceffectsof epidural osthole application on the mechanical allodyn-ia and the ERK/MAPK signaling pathway and the expression of COX-2 mRNA in the spinal dorsal horn.Methods 125adultmaleSDratswererandomizedin-to five groups( n=25 each) :Blank, Sham, NP, Ost and vehicle. At postoperative day 6, 1mg/rat osthole 50 μl was injected epidurally into group Ost and the same volume of vehicle was given into group vehicle. The mechanical pain threshold was measured by 50%MWT at 1 day before operation and the 3 rd,6 th,7 th, 14 th,21 st day after operation. After the measurement of pain threshold on postoperative day 14 , the L4-6 segment of spinal dorsal horn was removed for determi-nation of the expression of ERK, pERK and COX-2 mRNAbyWesternblotandRT-PCR.Results Com-pared with blank group, the mechanical pain threshold was only down-regulated at day 1 after operation in sham group, the expression of pERK and COX-2 mR-NA in sham group showed no significant difference ( P>0. 05 ); the mechanical pain threshold was signifi-cantly down-regulated after operation in NP, Ost and vehicle groups( P<0. 05 ) and the expression of pERK and COX-2 mRNA was significantly increased ( P <0. 05). Compared with vehicle group, the pain thresh-old in Ost group was significantly increased after drug administration( P<0. 05 ) and the expression of pERK and COX-2 mRNA was significantly reduced ( P <0. 05 ) . The expression of ERK showed no significant difference among each group(P>0. 05). The correla-tion analysis on pERK1/2 and COX-2 mRNA revealed the Pearson correlation coefficient was 0 . 878 and 0 . 910 , suggesting a strong positive correlation between pERKandCOX-2mRNA.Conclusions Ostholead-ministrated in the early stage after surgery can alleviate the nucleus pulposus-induced radicular inflammatory pain probably by inhibiting the expression of pERK and COX-2 mRNA in spinal dorsal horn.

Objectlve To demonstrate the feasibility of three-dimensional(3D)spin-lattice relaxation time in the rotating frame(T1ρ)-weighted imaging of porcine patellar cartilage in vitro at 7.0 T and the measurement of T1ρ of agarose phantom and patellar cartilage.Methods All the MR Imaging experiments were performed on a 7.0 T Varian scanner using a 6.0-cm-diameter quadrature birdcage RF coil tuned to 300 MHz.A 3D spin-echo sequence with a self.compensating spin-lock pulse cluster was used to acquire 3D-T1ρ-weighted images.The time of spin-locking(TSL)was from 0 to 50 ms with an interval of 10 ms.Spin-lock power wag 440 Hz.3D-T1ρweighted imaging was done three times for 6 phantoms (concentration 1%t0 6%),as well as once for 8 porcine patellar cartilages in order to assess the reproducibility of this technique.Signal-to-noise ratio(SNR)was measured on the acquired images of both phantoms and patellar cartilages,which were tested for significance using Two-way ANOVA.The images were processed on Vnmr J workstation using home-built processing software to construct 3 D T1ρ maps.T1ρ values were calculated within manually drawn regions-of-interest(ROI),and differences between groups were tested for significance using analysis of variance(One-way ANOVA).Results T1ρ -weighted images with a shorter TSL had a higher SNR value,which measured between 48±8 and 95±8 in the global cartilage.Cartilage images had a higher SNR(TSL＜30 ms)compared to agarose phantoms and a lower SNR(TSL ＞30 ms)only compared to l%agarose phantorm T1ρ relaxation times in agarose phantoms increased as agarose concentrations decreased in global regions.The CV of T1ρ in agarose phantoms was less than 10%.Global and regional analyses of patellar cartilage T1ρ were 68.9±6.3 ms,80.7±12.8 ms,65.7±7.0 ms,82.4±7.7 ms,and 69.7±6.4 ms in the global cartilage,the superficial layer,the transitional layer,the deep layer,and the calcified layer,respectively.T1ρ in the superficial and deep layer was significantly higher than in the transitional,calcified layer and global cartilage(F=6.436,P＜0.05).Conclusions The present study demonstrates the feasibihty of 3D-T1ρ-weighted imaging of porcine patellar cartilage at 7.0 T with hish image quality.T1ρ maps can be used to quantify the laminar structures in 3D-T1ρ-weighted images of articular cartilage,which pave the way to evaluate early osteoarthritis and cartilage regeneration.

Objective In order to understand the chondrogenesis differentiation of mesenchymal stem cells in either hydrogel or pellet culture,we applied the two methods and reveal the possible mechanism and for further investigation.Methods In C3H10T1/2 chondrogenic differentiation,we apply extracellular matrix hydrogel mixed the cell suspensions of freshly prepared (including scaling chondroitin sulfate,sodium hyaluronate synthesis and cross-linking agent) co-culture system and high cell density pellet formed by centrifugation.Chondrogenic differentiation of C3H10T1/2 was induced by treatment with TGF-β3 (10 ng/ml),dexamethasone (100 nmol/L),ascorbic acid (50 ug/ml),1 ∶ 100 dilution ITS+Premix and high glucose-DMEM medium with 0.2 volume fraction fetal bovine serum.And high glucose-DMEM medium with 0.2 volume fraction fetal bovine serum is for control group.Histochemistry staining was utilized to identify extracellular proteoglycan and real-time PCR was performed to assess gene expression of SOX9,collagen Ⅱa1/Ⅹa1 and aggrecan for the 1st,2nd and 3rd week respectively.Results In the hydrogel model for 3 weeks chondrogenic differentiation,the expression of master transcription factor SOX9 was upregulated in both culture models.While the marker genes of collagen Ⅱa1 and collagen Ⅹa1 were all promoted in hydrogel culture,the aggrecan gene expression was peaked in pellet culture.In addition,immunocytochemistry analysis of the hydrogel and pellet for 3 week illustrated the expression of extracellular matrix and more obviously in the hydrogel model.Conclusion In compared with pellet culture,the MSCs in the hydrogel were more likely promoted chondrogenesis leading to the eventual expression of marker genes.And the hydrogel would be applied in regeneration of cartilage injury.

Many clinicians are very busy for clinical works to have no time taking into account scientific research, so that they are daunting the application of scientific research funds. However, when clinicians in the actual work can keep their strong curiosity for the unknown, not cling to the so-called classical point of view, develop a good habit of critical thinking, have the courage to question the unresolved clinical problems, as well as read the latest literature of their specialty regularly, they may find the breakthrough point of scientific research to carry out scientific research within our ability. The results obtained in the process of scientific research aim to provide new methods and new technologies to solve clinical problems. Therefore, we could get a better way to serve the patients through the integration of clinical practice and scientific research to improve the development of clinical medicine. This article will focus on the aspects of selecting scientific research topic, and raising, establishment and solution of scientific problems etc, so as to helping clinicians to successfully get the support of the scientific funds.

Objective: To analyze the correlations between intraoperative ultrasound and MRI metrics of the spinal cord in degenerative cervical myelopathy and identify novel potential predictive ultrasonic indicators of neurological recovery for degenerative cervical myelopathy. Materials and Methods: Twenty-two patients who underwent French-door laminoplasty for multilevel degenerative cervical myelopathy were followed up for 12 months. The Japanese Orthopedic Association (JOA) scores were assessed preoperatively and 12 months postoperatively. Maximum spinal cord compression and compression rates were measured and calculated using both intraoperative ultrasound imaging and preoperative T2-weight (T2W) MRI. Signal change rates of the spinal cord on preoperative T2W MRI and gray value ratios of dorsal and ventral spinal cord hyperechogenicity on intraoperative ultrasound imaging were measured and calculated. Correlations between intraoperative ultrasound metrics, MRI metrics, and the recovery rate JOA scores were analyzed using Spearman correlation analysis. Results: The postoperative JOA scores improved significantly, with a mean recovery rate of 65.0 ± 20.3% (p < 0.001). No significant correlations were found between the operative ultrasound metrics and MRI metrics. The gray value ratios of the spinal cord hyperechogenicity was negatively correlated with the recovery rate of JOA scores (ρ = -0.638, p = 0.001), while the ventral and dorsal gray value ratios of spinal cord hyperechogenicity were negatively correlated with the recovery rate of JOA-motor scores (ρ = -0.582, p = 0.004) and JOA-sensory scores (ρ = -0.452, p = 0.035), respectively. The dorsal gray value ratio was significantly higher than the ventral gray value ratio (p < 0.001), while the recovery rate of JOA-motor scores was better than that of JOA-sensory scores at 12 months post-surgery (p = 0.028). Conclusion For degenerative cervical myelopathy, the correlations between intraoperative ultrasound and preoperative T2W MRI metrics were not significant. Gray value ratios of the spinal cord hyperechogenicity and dorsal and ventral spinal cord hyperechogenicity were significantly correlated with neurological recovery at 12 months postoperatively.

Objective: To analyze the correlations between intraoperative ultrasound and MRI metrics of the spinal cord in degenerative cervical myelopathy and identify novel potential predictive ultrasonic indicators of neurological recovery for degenerative cervical myelopathy. Materials and Methods: Twenty-two patients who underwent French-door laminoplasty for multilevel degenerative cervical myelopathy were followed up for 12 months. The Japanese Orthopedic Association (JOA) scores were assessed preoperatively and 12 months postoperatively. Maximum spinal cord compression and compression rates were measured and calculated using both intraoperative ultrasound imaging and preoperative T2-weight (T2W) MRI. Signal change rates of the spinal cord on preoperative T2W MRI and gray value ratios of dorsal and ventral spinal cord hyperechogenicity on intraoperative ultrasound imaging were measured and calculated. Correlations between intraoperative ultrasound metrics, MRI metrics, and the recovery rate JOA scores were analyzed using Spearman correlation analysis. Results: The postoperative JOA scores improved significantly, with a mean recovery rate of 65.0 ± 20.3% (p < 0.001). No significant correlations were found between the operative ultrasound metrics and MRI metrics. The gray value ratios of the spinal cord hyperechogenicity was negatively correlated with the recovery rate of JOA scores (ρ = -0.638, p = 0.001), while the ventral and dorsal gray value ratios of spinal cord hyperechogenicity were negatively correlated with the recovery rate of JOA-motor scores (ρ = -0.582, p = 0.004) and JOA-sensory scores (ρ = -0.452, p = 0.035), respectively. The dorsal gray value ratio was significantly higher than the ventral gray value ratio (p < 0.001), while the recovery rate of JOA-motor scores was better than that of JOA-sensory scores at 12 months post-surgery (p = 0.028). Conclusion For degenerative cervical myelopathy, the correlations between intraoperative ultrasound and preoperative T2W MRI metrics were not significant. Gray value ratios of the spinal cord hyperechogenicity and dorsal and ventral spinal cord hyperechogenicity were significantly correlated with neurological recovery at 12 months postoperatively.