Objective To observe the effect of BMSCs transplantation on gene and protein expression of VEGF receptor fetal l iver kinase 1 (Flk-1) after spinal cord injury (SCI), and to investigate the mechanism of repairing the SCI by BMSCs transplantation. Methods BMSCs were isolated and cultured from five 4-week-old male Wistar rats weighing100-120 g. The SCI model was made by using the modified Allen’s impactor device. Eighty-one adult female Wistar rats weighing 220-250 g were randomly divided into 3 groups: sham-operated group (group A, n=21), in which spinous process and vertebral plate of thorax 8-10 spinal cord segment were removed; DMEM group (group B, n=30), in which rats received four injections of DMEM in the peri-lesion area; and BMSCs group (group C, n=30), in which rats received four injections of BMSCs in the peri-lesion area. The changes of Flk-1 mRNA expression in rats’ spinal cord tissues were detected with RT-PCR method 1, 3 and 5 days after transplantation. The expression of Flk-1 protein was observed by using immunohistochemical technology in spinal cord 3, 7 and 14 days after transplantation. Results Morphology of the primary cultured BMSCs was various. Cell morphology tended to be uniform with the accumulation of passages, which appeared flat and spindle-shaped. RT-PCR results showed that there was no significant differences (P gt; 0.05) in Flk-1 mRNA expression between group C and group B at different time points after transplantation. But Flk-1 mRNA levels of group B and group C significantly increased and peaked 1 day after transplantation (P lt; 0.01), and then decreased 3 days after transplantation (P lt; 0.01) compared with that of group A, and were still higher than that of group A 5 days after transplantation (P lt; 0.05). Immunohistochemical staining results revealed that the expression of Flk-1 in group B was enhanced 3 and 7 days after transplantation compared with group A, which was significantly different (P lt; 0.01). There was no significant difference in the expression of Flk-1 between group B and groupp A 14 days after transplantation (P gt; 0.05). There was no significant difference in Flk-1 protein expression between group C and group B 3 days after transplantation (P gt; 0.05). The expression of Flk-1 protein in group C was significantly higher than that in group B 7 and14 days after transplantation (P lt; 0.01). Conclusion BMSCs transplantation after SCI does not have regulatary effect onthe expression of Flk-1 mRNA, but it does upregulate the Flk-1 protein expression, which may be one of the mechanisms of repairing SCI.
Objective To study the anatomy and variations of hepatic veins draining into inferior vena cava （IVC）， and to classify the surgical techniques of piggyback liver transplantation （PBLT） based on the view of hepatic veins anatomy with IQQA liver image analysis system so as to provide the important basis for the perioperative clinical decision making. Methods Two hundred and forty-eight cases of PBLT were preformed in the Zhongnan Hospital of Wuhan University and the 3rd Xiangya Hospital of Central South University from May 2000 to August 2007， the types of hepatic veins were summarized according to the anatomy of hepatic veins and short hepatic veins draining into IVC at the second and third hepatic hilars. Forty cases of PBLT were preformed in the Zhongnan Hospital of Wuhan University from March 2010 to April 2013， and the anatomy of hepatic veins was reviewed with IQQA liver image analysis system. The anatomy of hepatic veins and technological type of liver transplantation were recorded respectively. Results Of these 248 livers studied in our center， type Ⅰ（the left and middle hepatic vein joined as one trunk ） was found in 142 cases （57.25%）， type Ⅱ （the right and middle hepatic vein joined as one trunk） was 54 cases （21.77%）， type Ⅲ （three hepatic veins joined as one trunk） in 14 cases （5.64%）， type Ⅳ （the left， middle， and right hepatic veins were all unique）in 34 cases （13.71%）， and type Ⅴ （no hepatic veins but short hepatic veins） in 4 cases （1.61%）. The data of 40 cases of PBLT from IQQA liver image analysis system showed that type Ⅰwere found in 24 cases （60.00%）， type Ⅱin 9 cases（22.50%）， type Ⅲ in 2 cases （5.00%）， type Ⅳ in 4 cases （10.00%）， and type Ⅴ in 1 case （2.50%）， which were matched with hepatic vein classification standard of the author. Conclusions Studying the anatomy and variations of hepatic veins draining into IVC with IQQA liver image analysis system and classifying the surgical techniques of PBLT （type Ⅰ，Ⅱ，Ⅲ，andⅣA patients can be performed classical PBLT；Type ⅣB and Ⅴ patients can only be performed ameliorative PBLT） could provide an important basis for clinical preoperative decision.
ObjectiveTo explore the effectiveness of patellar tendon reconstruction by using LARS artificial ligament in treatment of old patellar tendon rupture.MethodsA clinical data of 12 patients with old patellar ligament ruptures, who met the inclusive criteria and reconstructed with LARS artificial ligament between December 2011 and December 2017, was retrospectively analyzed. There were 8 males and 4 females with an average age of 33.5 years (range, 18-55 years). The cause of injury included traffic accident injury in 4 cases, sport injury in 5 cases, and violent injury in 3 cases. There were 5 cases in the left knee and 7 cases in the right knee. The disease duration was 2-12 weeks (mean, 2.5 weeks). The preoperative Lysholm score and Kujala score were 43.2±3.2 and 43.9±2.6, respectively. The knee range of motion was (106.5±14.7)°. The thigh circumference which was measured at 10 cm above the upper end of the patella was (40.92±1.93) cm. There were 4 cases of patellar ligament body rupture, 1 case of patella distal pola rupture, and 7 cases of tibial tuberosity attachment rupture. Preoperative Caton-Deschamps index was 1.47±0.13.ResultsAll patients were followed up 12-30 months (mean, 20.5 months). All incisions healed by first intention. And no complication such as infection, recurrent rupture, and neurovascular injury occurred. At 1 year after operation, the knee range of motion was (131.0±10.2)°, Lysholm score was 87.4±2.4, Kujala score was 88.3±4.8, the thigh circumference which was measured at 10 cm above the upper end of the patella was (42.58±1.93) cm; all showing significant differences when compared with preoperative values (P<0.05). The effectiveness results were excellent in 9 cases and good in 3 cases according to the Insall evaluation criteria. The Caton-Deschamps index was 1.09±0.11, which was significantly lower than preoperative one (t=8.155, P=0.000).ConclusionPatellar tendon reconstruction with LARS artificial ligament is an effective method for the old patellar ligament rupture, which can effectively repair the knee extension device and restore knee function.
Objective To explore the protective effect and mechanism of Astragalus polysaccharides (APS) on liver injury in the state of brain death in New Zealand rabbits. Methods Twenty-four New Zealand rabbits were randomly divided into 3 groups (n=8): the blank control group, the brain death group, and the APS group. We obtained blood and liver tissue specimens from rabbits of three groups at 4 h and 8 h after treatment respectively (n=4). The rabbits of blank control group simulated the procedures of anesthesia and surgery of the brain death, without the Foley balloon catheter being pressurized, and maintained anesthesia. The brain death group: brain-dead models were established. The APS group: injection of APS (12 mg/kg) via the femoral vein bolus immediately after anesthesia, brain-dead models were established as same as rabbits of brain death group. The blood and liver tissue samples were taken at 4 h and 8 h after treatment to detect aminotrans-ferase (AST), alanine amino-transferase (ALT) and tumor necrosis factor α (TNF-α), and to observe the change of liver tissue by HE staining and immunohistochemical staining〔expression level of nuclear transcription factor p65 protein (NF-κB p65) could be detected by immunohistochemical staining〕. Results ① ALT and AST. Compare with the blank control group at the same time (4 h and 8 h), levels of ALT and AST in brain death group and APS group were significantly increased (P<0.05), and the levels of ALT and AST in brain death group were higher than those of APS group at each time point (P<0.05). In the same group, compared with 4 h, there was no significant difference in the levels of ALT and AST in blank control group at 8 h (P>0.05); the levels of ALT and AST in brain death group at 8 h were both higher than those of 4 h (P<0.05); the levels of ALT at 8 h in APS group was higher than that of 4 h, but there was no significant difference in the level of AST between 4 h and 8 h (P>0.05). ② TNF-α. Compare with the blank control groups at same time (4 h and 8 h), levels of TNF-α in brain death group and APS group were significantly increased(P<0.05), and level of TNF-α in brain death group was higher than that of APS group at 4 h and 8 h (P<0.05). ③ The HE results. The liver tissue structure of blank control group, brain death group, and APS group at 4 h had no obvious change. The liver tissue structure of brain death group at 8 h showed the evident tissue damage: liver cells showed the balloon samples, disordered arrangement, cytoplasmic loose light dye net-like, and inflammatory cells infiltrated in portal area. The liver tissue structure of APS group at 8 h showed that, liver cells showed mild edema, normal arrangement, and a small amount of inflammatory cells infiltrated in portal area. The liver tissue structure damage of APS group at 8 h was milder than that of brain death group. ④ Immunohistochemical staining results. There was no significant difference in expression levels of NF-κB p65 protein among blank control group, brain death group, and APS group at 4 h (P>0.05). But at 8 h, the expression levels of NF-κB p65 protein in brain death group and APS group were higher than that of blank control group (P<0.05), and the expression level of NF-κB p65 protein in brain death group was higher than that of APS group (P<0.05). The expression levels of NF-κB p65 protein in brain death group and APS group at 8 h was higher than that of 4 h in the same group (P<0.05), but there was no significant difference between 4 h and 8 h in blank control group (P>0.05). Conclusions Brain death will cause liver damage and the injury degree may be related to the continuous time. The damage at 8 h was more serious than that of 4 h. APS has a protective effect on liver of brain-dead rabbits' and its mechanism may be closely related to inhibit TNF-α and NF-κB by diverse ways to reduce the inflammation of the liver injury.