Research on Runx2 gene induced differentiation of human amniotic mesenchymal stem cells into ligament fibroblasts in vitro and promotion of tendon-bone healing in rabbits / 中国修复重建外科杂志

OBJECTIVE@#To investigate whether the Runx2 gene can induce the differentiation of human amniotic mesenchymal stem cells (hAMSCs) to ligament fibroblasts in vitro and promote the tendon-bone healing in rabbits.@*METHODS@#hAMSCs were isolated from the placentas voluntarily donated from healthy parturients and passaged, and then identified by flow cytometric identification. Adenoviral vectors carrying Runx2 gene (Ad-Runx2) and empty vector adenovirus (Ad-NC) were constructed and viral titer assay; then, the 3rd generation hAMSCs were transfected with Ad-Runx2 (Ad-Runx2 group) or Ad-NC (Ad-NC group). The real-time fluorescence quantitative PCR and Western blot were used to detect Runx2 gene and protein expression to verify the effectiveness of Ad-Runx2 transfection of hAMSCs; and at 3 and 7 days after transfection, real-time fluorescence quantitative PCR was further used to detect the expressions of ligament fibroblast-related genes [vascular endothelial growth factor (VEGF), collagen type Ⅰ, Fibronectin, and Tenascin-C]. The hAMSCs were used as a blank control group. The hAMSCs, hAMSCs transfected with Ad-NC, and hAMSCs were mixed with Matrigel according to the ratio of 1 : 1 and 1 : 2 to construct the cell-scaffold compound. Cell proliferation was detected by cell counting kit 8 (CCK-8) assay, and the corresponding cell-scaffold compound with better proliferation were taken for subsequent animal experiments. Twelve New Zealand white rabbits were randomly divided into 4 groups of sham operation group (Sham group), anterior cruciate ligament reconstruction group (ACLR group), anterior cruciate ligament reconstruction+hAMSCs transfected with Ad-NC-scaffold compound group (Ad-NC group), and anterior cruciate ligament reconstruction+hAMSCs transfected with Ad-Runx2-scaffold compound group (Ad-Runx2 group), with 3 rabbits in each group. After preparing the ACL reconstruction model, the Ad-NC group and the Ad-Runx2 group injected the optimal hAMSCs-Matrigel compunds into the bone channel correspondingly. The samples were taken for gross, histological (HE staining and sirius red staining), and immunofluorescence staining observation at 1 month after operation to evaluate the inflammatory cell infiltration as well as collagen and Tenascin-C content in the ligament tissues.@*RESULTS@#Flow cytometric identification of the isolated cells conformed to the phenotypic characteristics of MSCs. The Runx2 gene was successfully transfected into hAMSCs. Compared with the Ad-NC group, the relative expressions of VEGF and collagen type Ⅰ genes in the Ad-Runx2 group significantly increased at 3 and 7 days after transfection ( P<0.05), Fibronectin significantly increased at 3 days ( P<0.05), and Tenascin-C significantly increased at 3 days and decreased at 7 days ( P<0.05). CCK-8 detection showed that there was no significant difference ( P>0.05) in the cell proliferation between groups and between different time points after mixed culture of two ratios. So the cell-scaffold compound constructed in the ratio of 1∶1 was selected for subsequent experiments. Animal experiments showed that at 1 month after operation, the continuity of the grafted tendon was complete in all groups; HE staining showed that the tissue repair in the Ad-Runx2 group was better and there were fewer inflammatory cells when compared with the ACLR group and the Ad-NC group; sirius red staining and immunofluorescence staining showed that the Ad-Runx2 group had more collagen typeⅠ and Ⅲ fibers, tending to form a normal ACL structure. However, the fluorescence intensity of Tenascin-C protein was weakening when compared to the ACLR and Ad-NC groups.@*CONCLUSION@#Runx2 gene transfection of hAMSCs induces directed differentiation to ligament fibroblasts and promotes tendon-bone healing in reconstructed anterior cruciate ligament in rabbits.

Role of a continuously developed information traceability system in the management of foreign medical instrument packages / 中国基层医药

Objective:To investigate the role of a continuously developed information traceability system in the management of foreign medical instrument packages in central sterile supply department (CSSD).Methods:A total of 350 foreign medical instrument packages processed by CSSD during May to June 2021 in Shenzhen Hospital, University of Chinese Academy of Sciences were included in the control group. These packages were managed with the original information traceability system. A total of 375 foreign medical instrument packages processed by CSSD during July to August 2021 in Shenzhen Hospital, University of Chinese Academy of Sciences were included in the observation group. These packages were managed with a continuously developed information traceability system. Step-by-step reminding and early warning control were performed according to the problems encountered in actual operation. Quality management indexes of foreign medical instrument packages and the incidence of adverse events were compared between the two groups.Results:The percentages of good cleaning and functional integrity of the foreign medical instrument packages, functional integrity of sterile barrier after sterilization, timely monitoring of biological culture results, standardized handover, postoperative cleaning and detoxification in the observation group were 96.0% (360/375), 98.7% (370/375), 99.5% (373/375), 98.9% (371/375), and 99.7% (374/375), which were significantly higher than those in the control group [84.2% (295/350), 92.6% (325/350), 91.4% (320/350), 89.1% (312/350), 84.9% (297/350), χ2 = 28.48, 15.40, 27.72, 31.80, 58.12, all P < 0.05]. The percentages of instrument breaking, instrument mixed or damaged, wet packages, information label error, oversized and overweighed packages in the observation group were 0.8% (3/375), 0.3% (1/375), 0.8% (3/375), 0.3% (1/375), and 0.5% (2/375), respectively, which were significantly lower than those in the control group [4.3% (15/350), 4.6% (16/350), 3.4% (12/350), 10.0% (35/350), 7.7% (27/350), χ2 = 9.08, 14.65, 6.17, 36.34, 24.31, all P < 0.05]. Conclusion:A continuously developed information traceability system is very important for management of medical instrument processed in CSSD. A developmental traceability system not only continuously improves quality control of foreign medical instrument and decreases the incidence of adverse events, but also increases the responsibility and enthusiasm of staff, thereby ensuring the safety for patients.

Evaluation of knee stability after simulated reconstruction of the core ligament of the posterolateral complex / 中国组织工程研究

BACKGROUND: Injury of the posterolateral complex of the knee joint is a common type of multiple ligament injuries of the knee joint. The reconstruction of the posterolateral complex can restore the posterior and lateral stability of the knee joint and rebuild the stability of the knee joint. OBJECTIVE: To discuss the feasibility and clinical effect of modified LaPrade method for functional reconstruction of posterolateral complex of knee with autograft peroneus longus tendon. METHODS: Fourteen patients with posterolateral complex and posterior cruciate ligament injuries who were treated in the Department of Orthopedics, Affiliated Hospital of Zunyi Medical University from October 2014 to March 2017 were enrolled in this study. Posterior cruciate ligament and posterolateral complex were simultaneously constructed in stage one. The injury of the posterolateral complex of the knee joint was Fanelli type C. Modified LaPrade method was used to functional reconstruction of posterolateral complex of knee with peroneus longus tendon. The anatomy and function of the core ligament of the posterolateral complex was simulated. Follow-up time was beyond 1 year. The tibia posterior displacement on stress radiographs, lateral compartment gapping on varus stress radiographs at 0° knee extension, and external rotation angle of tibia at 30° knee flexion were compared before and after surgery. The joint function was evaluated according to the score of International Knee Documentation Committee and Lysholm Knee score. RESULTS AND CONCLUSION: (1) All patients were followed up for 12-18 months. All patients had no knee-length restriction, with flexion limitation in some patients. (2) At the last follow-up, the tibia posterior displacement on stress radiographs, lateral compartment gapping on varus stress radiographs and external rotation angle of tibia at 30° knee flexion were reduced from preoperation, with statistically significant differences (P=0.000). (3) The International Knee Documentation Committee function was corrected from D preoperatively to A in 8 cases and B in 6 cases postoperatively. The average Lysholm score was increased from (32.4±5.6) preoperatively to (82.7±6.4) postoperatively, and the differences were statistically significant (P=0.000). (4) It is indicated that with peroneus longus tendon, the anatomy and function of the core ligament of the posterolateral complex were simulated by modified LaPrade method to functional reconstruction of posterolateral complex of knee, and the postoperative knee function recovered well.