Objective To investigate mRNA and protein expressions of TGF-β1 and their signifi-cance in injured spinal cord after transplantation of activated microglia. Methods Wistar rat model of spinal cord injury was made by using modified Allen's method. Then, the activated microglia was trans-planted into spinal cord injury models tu detect mRNA and protein expressions of TGF-β1 in the spinal curd. Results The mRNA and protein expressions of TGF-β1 in control group were significantly lower than that in test group (P<0.05). Conclusion Transplantation of activated microglia to the impaired spinal cord in rats can increase the mRNA and protein expressions of TGF-β1. The up-regulated expres-sion of TGF-β1 in spinal cord may play a role in promoting the recovery of spinal cord, when the activated microglia may be the main saurce of TGF-β1.

Objective To develop a new scoring system,Daping orthopedics operation risk scoring system for senile patient(DORSSSP),for preoperative evaluation of operative risks in the elderly patients with hip fractures based on acute physiology and chronic health evaluation(APACHE)Ⅱ scoring system and physiological and operative severity score for the enumeration of mortality and morbidity (POSSUM)and compare the new scoring system with APACHE Ⅱ and POSSUM in assessing surgical risks and predicting postoperative complications and mortalities.Methods A total of 260 patients with hip fractures treated in our department in recent five years were retrospectively and respectively evaluated with DORSSSP,POSSUM,progressed POSSUM(P-POSSUM)and APACHE Ⅱ scoring system to compare the value of three scoring systems in preoperative evaluation of operative risks and prediction of postoperative mortality and complications.Results POSSUM and DORSSSP predicted complications in 119 and 92 patients respectively,while the actual complication occurred in 84 patients.The prediction value of POSSUM was significantly higher than the actual value,while the prediction value of DORSSSP showed no statistical difference compared with the actual value.POSSUM,P-POSSUM and APACHEⅡ scoring systems predicted 16,10 and 12 deaths respectively,but there were six deaths in fact,with prediction value obviously higher than the actual value.DORSSSP predicted nine deaths,the closest value to the actual.Conclusions DORSSSP has good correlation with postoperative complications and mortalities.Compared with POSSUM and APACHE Ⅱ scoring system,more simple and practicable DORSSSP can more accurately evaluate the preoperative risks and predict the postoperative complications and mortalities in the elderly patients with hip fractures.

Objective To observe the anatomical and histological features of anterolateral ligament (ALL)in the knee of Chinese adults,so as to identify the existence of ALL and provide an anatomical foundation for clinical reconstruction.Methods Ten adult knee specimens were randomly selected to be dissected,and the femoral,tibial and meniscus attachment points of the ALL were observed.The length,width and thickness were measured using the vernier caliper after the dissection.Three specimens were subjected to histological staining in the end.Results (1)ALL originated from the lateral femoral condyle—the same point of the lateral collateral ligament femoral side or the distal-anterior side,with its body divided into two branches,located in the tibia and the lateral meniscus respectively.The starting point of tibial side ALL was located at the mid-point of Gerdy's tubercle to fibula head,below tibial cartilage edge,with the meniscus point located in the lateral meniscus anterior horn and body junction area.(2) The average length of ALL is 38.89 ± 4.67 mm.The width in the femur,tibial attachment point was fan-shaped spread connected with sclerotin,being the narrowest at the joint line.The width at the femur,tibial attachment point and the joint line was 8.49 ± 1.36 mm,8.15 ± 1.38 mm and 6.49 ± 1.09 mm respectively,with the thickness of 1.33 ± 0.38 mm.The distance from tibia attachment points to the Gerdy's tubercle,fibular head and tibia cartilage margin was 22.59 ± 3.04 mm,21.15 ± 2.78 mm and 5.76 ± 0.57 mm respectively.(3) HE staining showed that ALL was dense connective tissue consisting of parallel arranged collagen fibers,while S-100 staining indicated that ALL contained sensory motor nerve fibers.Conclusion ALL is independent of the joint capsule and originates from the femoral lateral condyle.Its body is divided into two branches,located in the tibia and the lateral meniscus respectively.

BACKGROUND:In recent years, the number of cases of anterior cruciate ligament reconstruction with remnant preservation is increased year by year, but its clinical results, especialy effects on improving proprioceptive recovery after reconstruction, are stil controversial. OBJECTIVE: To compare the clinical effects of arthroscopic anterior cruciate ligament reconstruction with or without remnant preservation. METHODS:Totaly 146 patients undergoing arthroscopic anterior cruciate ligament reconstruction were randomly divided into two groups; preserving-remnant group and removing-remnant group. Autologous hamstring tendons were selected. Evaluation of knee mobility, Lysholm score, IKDC scores and knee stability was performed before and 6, 12 months after reconstruction. Proprioception was recorded before and 3, 6 and 12 months after reconstruction. The comparative analysis was carried out on these data between the group and between affected and healthy limbs. RESULTS AND CONCLUSION:114 patients were folowed up for over 12 months, including 61 in the preserving-remnant group and 53 in the removing-remnant group. There were significantly statistical improvements in knee mobility, Lysholm score, IKDC score and knee stability at 6 and 12 months after reconstruction in the two groups (P < 0.01), and the Lysholm score, IKDC score and knee stability were better in the removing-remnant group than the removing-remnant group at 6 months after reconstruction (P > 0.05). The knee proprioception was significantly improved at 3, 6 and 12 months after reconstruction in the two groups (P < 0.01), and it was also better in the removing-remnant group than the removing-remnant group at 3 and 6 months after reconstruction (P < 0.05). These findings indicate that the anterior cruciate ligament reconstruction with remnant preservation is beneficial to the recovery of postoperative proprioception and knee function.

BACKGROUND: FG loop (FGL) is a core active peptide fragment of neural cell adhesion molecule (NCAM), which can directly act on fibroblast growth factor receptor 1 (FGFR1) to activate NCAM signal pathway.OBJECTIVE: To observe the effects of synthetic peptides FGL on PC12 cells proliferation and apoptosis.METHODS: ①PC12 cells proliferation and apoptosis: The cultured PC12 cells were divided into control group and experiment group. The experimental group was added with 1% FGL peptide solution. The control group was pre-coated with poly-lysine plates. The cells were cultured 1, 3, 5, 7, 9 d respectively to detect cell proliferation by using Cell Counting Kit-8. ②PC12 apoptosis and nuclear factor kappa B mRNA detection: The PC12 cells were divided into normal group, experimental group and injury group. H2O2 was added into the injury group for 16 hours stimulation. In the experimental group, H2O2 and FGL were used for 16 hours stimulation. The cell apoptosis were detected by flow cytometry; mRNA expression of nuclear factor kappa B was detected by quantitative fluorescent polymerase chain reaction.RESULTS AND CONCLUSION: PC12 cells cocultured with FGL peptide grow well, which indicates that FGL peptides can promote PC12 cell proliferation and inhibit PC12 cell apoptosis, as well as decrease mRNA expression of nuclear factor kappa B.

BACKGROUND:Bacterial biofilm is the main cause of the infection of the prosthesis.In vitro experiments confirmed that hypertonic sodium chloride and ethanol can apparently promote the formation of staphylococcal biofilms. There are no reports on the effects of ethanol and hypertonic environment surrounding the prosthesis on the formation of biofilms.
OBJECTIVE: To evaluate the effects of different environment factors surrounding the prosthesis on the growth of Staphylococcus epidermidis and bacterial biofilm formation after replacement.
METHODS: White rabbit models infected with Staphylococcus epidermidis on the prosthesis were established, and were randomly divided into hypertonic sodium chloride, ethanol and control groups (n=15). The bacteria were injected with 0.1 mL 4% sodium chloride and 4% ethanol into the knee of rabbits in the hypertonic sodium chloride and ethanol groups. The rabbits were injected with 0.1 mL 0.9% sodium chloride in the control group. Three rabbits were sacrificed at 2, 4, 6, 8 and 16 days after inoculated with bacteria. Synovial fluid, prosthesis and tissue surrounding infection were obtained. Bacterium was cultured to extract total RNA. The ica operon transcription levels were detected in the gene levels. Adhesion of bacteria on the surface of the prosthesis was observed using a scanning electron microscope. Tissues surrounding the prosthesis were observed using hematoxylin-eosin staining.
RESULTS AND CONCLUSION:Histological examination revealed that inflammatory cel infiltration was observed in al the rabbits at 4 days after injection. Colony formation was found at 16 days after injection. At 6 days after injection, inflammatory cel infiltration was observed in the ethanol and control groups. Scanning electron microscope showed that compared with the control group, the bacteria adhered to the prosthetic surface became more in the hypertonic sodium chloride and ethanol groups at 6, 8 and 16 days (P < 0.05). At 6, 8 and 16 days, the expression of icaA mRNA was significantly higher in the hypertonic sodium chloride and ethanol groups than in the control group (P < 0.05). These data showed that the environment factors could affect the growth of Staphylococcus epidermidis and bacterial biofilm formation.

Objective To study the role of activated microglia transplantation in recovery of hindlimb locomotor function in rats with spinal cord injury(SCI). Methods A total of 40 female adult Wistar rats were selected and divided into four groups randomly(10 rats in each group).The former two groups were locomotor function observation groups,in which rat model with spinal cord was established by striking with improved self-made Allen's strike equipment to fabricate moderate spinal cord injury and divided into transplantation group and control group.The latter two were histological observation groups,the spinal cord injury model was fabricated by the same above-mentioned method and divided into transplantation and control groups.Before fabricating the spinal cord injury model,the microglia of the newborn rats were cultured,separated,purified and identified and the purity of the microglia determined.The injury position was exposed again seven days after transplantation and the cell suspension of microglia was injected around the injury position with microsyringe,which was free in the control group.The hindlimb locomotor function of rats was detected and scored at 1 day,1,2,3 and 4 weeks in the locomotor function observation groups after transplantation respectively.At the same time,two rats were extracted randomly from the control group and the transplantation group in histological observation groups to cut specimen and slice for Naoumenko-Feign paraffin section and dying.Then,the microglia were observed and counted by microscope and analyzed statistically. Results At 1,2,3 and 4 weeks after operation,the BBB score of the control group and the transplantation group was increased gradually with the time.But compared with the control group,transplantation group had higher scores of hindlimb locomotor function at 2,3 and 4 weeks after operation,with statistical difference(P＜0.05).Naoumenko-Feigin paraffin section and dying and microglia counting showed that positive microglia number in the transplantation group was increased more obviously than the control group,with statistical difference(P＜0.05).Conclusion Activated microglia transplantation can promote the recovery of the hindlimb locomotor function in SCI rats.

Objective:To investigate the repair effects and mechanism of mesenchymal stem cell osteogenesis-derived exosomes deliver miRNA-222-5p on tendon cell injury.Methods:Mesenchymal stem cell exosomes were collected, isolated and characterized. The mice achilles tendon was collected after cutting one week later. The tendon cells were isolated and cultured to obtain a tendon cell injury model (injury group). The subjects were divided into three groups. During the process of osteogenic differentiation, mesenchymal stem cells were co-cultured with tendon cells (co-culture group). Further, miRNA-222-5p mimics were transfected into mesenchymal stem cells, the co-cultured mesenchymal stem cells and tendon cells during the process of osteogenic differentiation (miRNA-222-5p group). Clone formation and Transwell were used to detect the ability of mesenchymal stem cells to secrete exosomes and exosomes transported miRNA-222-5p to repair damaged tendon cells. qPCR, Western-blot and fluorescence staining were used to detect the expression levels of cartilage-related proteins and bone-related proteins on mRNA and protein. Western-blot was used to detect the expression of signal pathway factors.Results:The shape of the exosomes was a typical cup - like structure. The exosome proteins marker was all expressed positively ( P<0.05). After counting the colony formation and Transwell test, the number of cell communities and cell invasion were significantly increased in the co-culture group compared with those in the injury group. The number of those in miRNA-222-5p group were significantly increased compared with those in co-culture group ( P<0.05). The qPCR, Western-blot and fluorescent staining test results showed that the mRNA and protein expression levels of SOX-9, COL2A1, ACAN, BMP2, Runx2, and OPN in the co-culture group were significantly increased compared with the injury group. The mRNA and protein expression levels of the above proteins in the miRNA-222-5p group were significantly increased compared with those in co-culture group ( P<0.05). We also detected the protein expression levels of signal pathway related factors. Western-blot results showed that the expression levels of NF-κB, Erk2, STAT3, STAT5 and Smad2 in the co-culture group were significantly increased compared with those in the injury group. The expression levels of the above factors in the miRNA-222-5p group were significant increased compared with the co-culture group ( P<0.05). Conclusion:The co-culture of mesenchymal stem cells and achilles tendon cells could promote the osteogenic differentiation of achilles tendon cells. Exosomes derived from mesenchymal stem cells could further promote the osteogenic differentiation of achilles tendon cells by transporting miRNA-222-5p. The mechanism may be related to the up-regulation of signal pathway factors in injured tendons, including NF-κB, Erk2, STAT3, STAT5 and Smad2.

Objective To investigate the vascular endothelial growth factor (VEGF) expression level by chondrocytes isolated from patients with osteoarthritis (OA) in hip or femoral neck fracture (FNF) and explore the effect of synovial fluid from OA or FNF on secretion of VEGF. Methods The cartilage tissues were collected from 12 patients with OA in hip and 8 patients with FNF. Cartilage was stained with HIM and Safranin O/Fast Green (S/F) method. The damage of cartilage was evaluated using Mankin scores.Cathepsin B which was selected for cell dedifferentiation monitoring marker and VEGF level was detected in the supernatant fluid. The synovial fluid from OA, FNF and DMEM were respectively added to the culture medium to explore their effects on regulating VEGF. Results Cartilage the Mankin scores of OA group were higher than that of FNF group. Chondrocytes gradually lost their original spherical appearance, with Cathepsin B upregulated while VEGF downregulated. The OA synovial fluid can stimulate chongdrocytes to secrete more VEGF than the one from patients with FNF. However, chondrocytes gradually produced less VEGF after passaging. Conclusion Mankin scores had good correlation with chondrocytes' VEGF production in the early stage of primary culture. Chondrocytes showed quick dedifferentiation characteristics in vitro. OA synovial fluid showed abig ger capability in stimulating chondrocytes to express more VEGF, which might indicate that OA synovial fluid participated in the pathological process of OA.

Objective To compare the clinical outcomes of irradiated versus non-irradiated deepfrozen bone-patellar tendon-bone (B-PF-B) ullograft in anterior cruciate ligament (ACL) reconstruction. Methods A total of 66 patients undergoing arthroecopic ACL reconstruction were prospectively random-ized consecutively into two groups, ie, Group A ( irradiated deep-frozen allograft, n = 34) and Group B ( non-irradiated deep-frozen allograft, n = 32). All ACL reconstructions were done by the same senior surgeon with the same arthroscopic technique. Before and after surgery, the clinical results were compared in aspects of general conditions, range of motion ( ROM), Pivot shift test, Lachman and Anterior Drawer Test (ADT), Daniel's one-leg hop and Harner's vertical jump tests, overall international knee docu-mentation committee (IKDC) rating and KT-2000 arthrometer testing. Results Of all, 63 patients in-eluding 32 patients in Group A and 31 in Group B were available for a follow-up of average 38 months ( mean 38.3 months in Group A and mean 37.7 months in Group B) and three lost follow-up. There was one patient with late septic infection. While there was no statistical difference in aspects of general condi-tions including hospital stay, duration of fever and complications ( P > 0. 05 ), but there was a trend that the patients in Group A had a longer postoperative duration of fever ( mean 8.9 days ) than Group B (mean 7.8 days). Physical examinations showed no statistical difference upon ROM in both groups ( P > 0. 05), while there was statistical difference between Lachman test and ADT ( P < 0.05 ). The positive Pivot shift test was found in 12 patients in Group A and 3 in Group B, with statistical difference ( P < 0. 05 ). KT-2000 testing showed a side-to-side difference of less than 3 mm in 26 patients in Group B and only 8 in Group A, and a side-to-side difference of more than 5 mm in two patients in Group B and 12 in Group A, with statistical difference (P < 0.05 ). The anterior and rotational stability was decreased sig-nificantly in Group A. No statistical difference was found between two groups in overall IKDC rating, Daniel' s one-leg hop and Harner' s vertical jump tests ( P > 0.05 ). However, the function and IKDC score tended to decrease in Group A. Conclusion The short term clinical outcomes of the ACL reconstruction with irradiated B-PT-B allograft are adversely affected and unsatisfactory, indicating a cautious use.