Objective To analyze the stabil ity and cl inical outcomes of arthroscopic anterior cruciate l igament (ACL) reconstruction with γ irradiated patellar tendon allograft compared with autograft. Methods From January 2004 to October 2007, 69 patients undergoing arthroscopic ACL reconstruction were prospectively randomized consecutively into two groups: group A (autograft, n=36) and group B (γ irradiated allograft, n=33). In group A, there were 30 males and 6 females with an average age of 30.1 years, including 30 cases of simple ACL rupture and 6 cases of ACL rupture with medial accessory l igament injury; ACL rupture was caused by sports in 28 cases, by traffic accident in 5 cases, and by others in 3 cases; and the time from injury to operation was 1.4 months on average. In group B, there were 26 males and 7 femaleswith an average age of 32.5 years, including 27 cases of simple ACL rupture and 6 cases of ACL rupture with medial accessory l igament injury; ACL rupture was caused by sports in 27 cases, by traffic accident in 4 cases, and by others in 2 cases; and the time from injury to operation was 1.5 months on average. There were no significant differences in general data between two groups (P gt; 0.05). The same arthroscopic technique was used in all ACL reconstructions done by the same surgeon. The cl inical outcome was evaluated and compared by general conditions, pivot shift test, Lachman test, KT-2000 arthrometer testing, Daniel’s one-leg hop test, International Knee Documental Committee (IKDC) scoring, Lysholm knee scoring scale, and Tegner activity score. Results All patients were followed up for 39.5 months (group A) and 37.6 months (group B). In group A, patella fracture occurred in 1 case and anterior knee pain in 2 cases postoperatively. No compl ication occurred in group B. The hospital ization times in groups A and B were (15.6 ± 2.4) days and (15.5 ± 1.5) days, respectively, showing no significant difference (P gt; 0.05). The operation time of group A was longer than that of group B and the fever time of group A was shorter than that of group B, showing significant differences (P lt; 0.05). At the final follow-up, there were significant differences (P lt; 0.05) in Lachman test and the pivot shift test between two groups, between pre- and post-operation; there were no significant differences (P gt; 0.05) in Daniel’s one-leg hop test, the IKDC, Lysholm, and Tegner activity scores between two groups, however, there was a decreased trend in the functional and activity levels in group B. And there was significant difference between pre- and post-operation (P lt; 0.05). At the final follow-up, the differences between normal side and affected side were (2.4 ± 0.6) mm in group A and (5.5 ± 3.6) mm in group B, showing significant difference (P lt; 0.05). There was significant difference in tibial advancement between pre- and post-operation (P lt; 0.05). Conclusion The functional and activity level of the knee after ACL reconstruction with autograft and γ irradiated patellar tendon allograft were similar, but anterior and rotational stabil ity of the involved knee decreases significantly in the group with γ irradiated patellar tendon allograft.
Objective To compare the efficacy of reteplase and ateplase in the treatment of acute massive pulmonary thromboembolism ( PTE) in emergency. Methods From January 2005 to December 2009,42 patients with acute massive PTE were treated by intravenous thrombolysis with reteplase or ateplase. The thrombolysis efficacy, bleeding incidence and mortality were measured. Results In the reteplase group, the emergency thrombolysis effective rate was 88. 9% among 18 patients. Mild bleeding occurred in 3 patients,moderate bleeding in 1 patient, and 2 cases died in hospital. In the ateplase group, the emergency thrombolysis effective rate was 75% among 24 patients. Mild bleeding occurred in 3 patients, moderate bleeding in 2 patients, and 3 cases died in hospital. The thrombolysis effective rate, bleeding incidence and mortality had no significant difference between the two groups. Conclusion Both the reteplase and ateplase thrombolysis therapy are safe and effective in the treatment of acute massive PTE, but reteplase thrombolysis therapy is more convenient in emergency.
Objective To study the effect of platelet lysate (PL) on chondrogenic differentiation of human umbil ical cord derived mesenchymal stem cells (hUCMSCs) in vitro. Methods Umbil ical cords were voluntarily donated by healthy mothers. The hUCMSCs were isolated by collagenase digestion and cultured in vitro. The surface markers of the cells were detected by flow cytometer. According to different components of inductive medium, the cultured hUCMSCs were divided into 3 groups: group A [H-DMEM medium, 10% fetal bovine serum (FBS), and 10%PL]; group B [H-DMEM medium, 10%FBS,10 ng/mL transforming growth factor β1 (TGF-β1), 1 × 10-7 mol/L dexamethasone, 50 μg/mL Vitamin C, and 1% insul intransferrin- selenium (ITS)]; and group C (H-DMEM medium, 10%FBS, 10 ng/mL TGF-β1, 1 × 10-7mol/L dexamethasone, 50 μg/ mL vitamin C, 1%ITS, and 10%PL). The hUCMSCs were induced in the mediums for 2 weeks. Toluidine blue staining was used to detect the secretion of chondrocyte matrix. Immunofluorescence method was used to identify the existence of collagen trpe II. The expressions of Aggrecan and collagen type II were detected by semiquantitative RT-PCR. Results Flow cytometer results showed that the hUCMSCs did not express the surface markers of hematopoietic cell CD34, CD45, and human leukocyte antigen DR, but expressed the surface markers of adhesion molecule and mesenchymal stem cells CD44, CD105, and CD146. Toluidine blue staining and immunofluorescence showed positive results in group C, weak positive results in group B, and negative results in group A. Semiquantitative RT-PCR showed the expressions of Aggrecan and collagen type II at mRNA level in groups B and C, but no expression in group A. The mRNA expressions of Aggrecan and collagen type II were higher in group C than in group B (P lt; 0.05). Conclusion Only 10%PL can not induce differentiation of hUCMSCs into chondrocytes, but it can be a supplement to the induced mediums. PL can improve hUCMSCs differentiating into chondrocytes obviously in vitro. This study provides new available conditions for constructing tissue engineered cartilage.
Objective To construct inducible lentiviral vector containing human bone morphogenetic protein 2 (hBMP-2) gene and to study its expression in human umbil ical cord blood mesenchymal stem cells (HUMSCs). Methods hBMP-2 gene was ampl ified by PCR from a plasmid and was cloned into pDown by BP reaction. pLV/EXPN2-Neo-TRE-hBMP-2 and pLV/EXPN2-Puro-EF1A-reverse transactivator (rtTA) were obtained with GATEWAY technology, and then were sequenced and analyzed by PCR. The recombinant vectors were transfected into 293FT cells respectively through l ipofectamine, and the lentiviral viruses were harvested from 293FT cells, then the titer was determined. Viruses were used to infect HUMSCs in tandem. In order to research the influence of induction time and concentration, one group of HUMSCs was induced by different doxycl ine concentrations (0, 10, 100 ng/mL, and 1, 10, 100 μg/mL) in the same induction time (48 hours), and the other by the same concentration (10 μg/mL) in different time points (12, 24, 48, and 72 hours). The expression of target gene hBMP-2 was indentified by ELISA method. After 2-week osteogenic induction of transfected HUMSCs, the mineral ization nodes were detected with Al izarin bordeaux staining method. Results Therecombinant inducible lentiviral vectors (pLV/EXPN2-Neo-TRE-hBMP-2 and pLV/EXPN2-Puro-EF1A-rtTA) were successfully constructed. The lentiviruses were also obtained and mediated by 293FT cells, and the virus titers were 3.5 × 108 TU/mL and 9.5 × 107 TU/mL respectively. HUMSCs could expression hBMP-2 by induction of doxycycl ine. The expression of hBMP-2 reached the peak at 10 μg/mL doxycl ine at 48 hours of induction. After 2-week osteogenic induction, a lot of mineral ization nodes were observed. Conclusion The recombinant inducible lentiviral vectors containing hBMP-2 gene can be successfully constructed, which provide an effective and simple method for the further study of stem cells and animal experiment in vivo.