ABSTRCT AIM:To verify the effect of polygona-tum polysaccharide(PSP)combined with TGF-β3 in inducing the differentiation of rat tendon-derived stem cells(TDSCs)into chondrocytes by activating the TGF-β3/Smad2 pathway.METHODS:TDSCs were extracted from rat tail tendons using enzyme digestion,purified,passaged,and identified via flow cytometry.TDSCs were treated with different concentrations of PSP,and the optimal growth con-centration was determined using the CCK-8 assay.TDSCs were divided into four groups:PSP,TGF-β3,PSP+TGF-β3,and Control for differentiation induc-tion.Chondrogenic differentiation was evaluated using morphological observations,toluidine blue staining,immunofluorescence staining,and West-ern blot analysis to detect COLⅡ,SOX9,and AGG.Western blot was also used to measure the expres-sion levels of Smad2 and p-Smad2 to evaluate the activation of the TGF-β3/Smad2 pathway after chondrogenic induction.RESULTS:Flow cytometry analysis showed that TDSCs highly expressed CD90 and CD29,while CD11b and CD45 expression was low.The CCK-8 assay indicated that 5 μmol/L PSP was the optimal intervention dose.Toluidine blue staining revealed that the blue staining area in the PSP,PSP+TGF-β3,and TGF-β3 groups was larger compared to the Control group.Immunofluores-cence analysis demonstrated that COLⅡ expression was significantly increased in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest expression in the PSP+TGF-β3 group(P＜0.05).Western blot analy-sis showed that the levels of p-Smad2/Smad2,COLⅡ,SOX9,and AGG were elevated in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest ex-pression in the PSP+TGF-β3 group(P＜0.05).Compared to the Control group,the TGF-β3 and PSP groups also showed significantly increased expression(P＜0.05).CONCLUSION:PSP promotes the proliferation and chondrogenic differentiation of TDSCs,possibly by activating the TGF-β3/Smad2 pathway.

ABSTRCT AIM:To verify the effect of polygona-tum polysaccharide(PSP)combined with TGF-β3 in inducing the differentiation of rat tendon-derived stem cells(TDSCs)into chondrocytes by activating the TGF-β3/Smad2 pathway.METHODS:TDSCs were extracted from rat tail tendons using enzyme digestion,purified,passaged,and identified via flow cytometry.TDSCs were treated with different concentrations of PSP,and the optimal growth con-centration was determined using the CCK-8 assay.TDSCs were divided into four groups:PSP,TGF-β3,PSP+TGF-β3,and Control for differentiation induc-tion.Chondrogenic differentiation was evaluated using morphological observations,toluidine blue staining,immunofluorescence staining,and West-ern blot analysis to detect COLⅡ,SOX9,and AGG.Western blot was also used to measure the expres-sion levels of Smad2 and p-Smad2 to evaluate the activation of the TGF-β3/Smad2 pathway after chondrogenic induction.RESULTS:Flow cytometry analysis showed that TDSCs highly expressed CD90 and CD29,while CD11b and CD45 expression was low.The CCK-8 assay indicated that 5 μmol/L PSP was the optimal intervention dose.Toluidine blue staining revealed that the blue staining area in the PSP,PSP+TGF-β3,and TGF-β3 groups was larger compared to the Control group.Immunofluores-cence analysis demonstrated that COLⅡ expression was significantly increased in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest expression in the PSP+TGF-β3 group(P＜0.05).Western blot analy-sis showed that the levels of p-Smad2/Smad2,COLⅡ,SOX9,and AGG were elevated in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest ex-pression in the PSP+TGF-β3 group(P＜0.05).Compared to the Control group,the TGF-β3 and PSP groups also showed significantly increased expression(P＜0.05).CONCLUSION:PSP promotes the proliferation and chondrogenic differentiation of TDSCs,possibly by activating the TGF-β3/Smad2 pathway.

Objective: To evaluate the clinical application of single-arm external stent combined with free flap in the treatment of forearm fractures of Gustilo type Ⅲ. Methods: A retrospective study was conducted of the 16 patients who had been treated at Repair and Reconstruction Center, Traditional Chinese Medicine Hospital Affiliated to Southwest Medical University from September 2015 to January 2018 for open forearm fractures combined with soft tissue defects with single-arm external stent combined with free flap. They were 11 men and 5 women, aged from 18 to 64 years (average, 41.6 years). By the Gustilo classification, 9 cases were type ⅢB and 7 type ⅢC. The area of soft tissue defects at the upper arm and hand ranged from 7.5 cm×5.5 cm to 16.5 cm × 11.0 cm. Emergency debridement was performed at the primary stage. After repair of major blood vessels, nerves and tendons, the reduced fractures were fixated with a single-arm external stent. The soft tissue defects were repaired with free flaps at the secondary stage. Nine cases were repaired with a free anterolateral perforating branch flap and 7 with a free ilioinguinal flap. The single-arm external stent became the ultimate fixation mode in 5 cases but was changed into plate fixation after survival of the flaps in the other 11 cases. Complications were recorded postoperatively. At the last follow-up, the upper limb function was evaluated according to the tentative criteria for evaluation of the upper limb function proposed by the Hand Surgery Society of Chinese Medical Association. Results: Of all the free flaps, 14 survived smoothly but 2 anterolateral ones survived only after the venous crisis appearing at 24 h after operation was relieved by exploration. The 16 patients were followed up for 9 to 18 months (average, 13.5 months). The fractures united well with fine alignment of the fracture ends and recovered force line. According to the Anderson criteria for forearm fractures, 10 cases were excellent, 4 good and 2 fair after operation. According to the tentative criteria for evaluation of the upper limb function proposed by the Hand Surgery Society of Chinese Medical Association, 11 cases were excellent and 5 good. No nail infection or nonunion occurred. Conclusion In the treatment of forearm fractures of Gustilo type Ⅲ, single-arm external stent plus free flap can effectively restore the force line of upper extremity, promote bone healing, allow reasonable timing for wound repair, reduce postoperative complications like infection and osteomyelitis and facilitate functional recovery of the affected extremity.

Objective To observe the effects of erythropoietin-α administered before the total knee arthroplasty (TKA) on the blood transfusion in patients with mild anemia. Methods 60 patients with mild anemia, who were arranged to receive TKA , from 2010-2012 , were assigned to received either 40 000U of erythropoietin-αfor 3 times (once a week) 3 weeks before the surgery (intervention group, n=30) or not (control group, n=30). Then the hemoglobin levels , transfusion ratio , inpatient length of stay and duration of surgery between the 2 groups were analyzed. Results The hemoglobin level was increased from 11.5 g/dL to 14.3 g/dL in intervention group before the surgery (P<0.01). The blood transfusion rate was decreased from 96.6%to 56.6% (P=0.013) during the surgery. And the length of hospital stay was similar between the 2 groups. Conclusion Erythropoietin-αadministered 3 weeks before the TKA surgery could reduce the blood transfusion rate of patients with mild anemia.