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BACKGROUND:EdU is a new nuclear marker, and currently, it is rarely reported. OBJECTIVE: To determine the optimal concentration of EdU to label human umbilical cord mesenchymal stem cels. METHODS: Human umbilical cord mesenchymal stem cels were isolated, purified and subcultured. Cel morphology and growth were observed under inverted microscope. Flow cytometry was used to identify cel surface markers, as wel as adipogenic identification. EdU at concentrations of 5, 10, 20, 50, 100 μmol/L was used to label human umbilical cord mesenchymal stem cels for 24 hours. The optimal concentration that resulted in the highest labeling efficiency was selected, and then cel proliferation curve was drawn. RESULTS AND CONCLUSION:Under the inverted microscope, cels grew adherently in a long spindle shape, and EdU-labeled cels had the same morphology. Flow cytometry showed that cels were positive for CD44, and had adipogenic differentiation ability. When the concentration of EdU was 5 and 10 μmol/L, the labeling efficiency was the highest, indicating that 5 and 10 μmol/L are the optimal concentrations of EdU to label human umbilical cord mesenchymal stem cels.
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BACKGROUND:Cold trypsin digestion is rarely reported to culture umbilical cord mesenchymal stem cells. OBJECTIVE:To compare the biological characteristics of umbilical cord mesenchymal stem cells cultured by cold trypsin digestion and tissue explant method. METHODS:Human umbilical cord mesenchymal stem cells were isolated, purified and passaged using cold trypsin digestion and tissue explant method, and then the first adhesion time and cellcycle were recorded. Morphology of umbilical cord mesenchymal stem cells was observed under inverted phase contrast microscope to draw growth curve of cells at passage 3. Flow cytometry was used to detect the surface markers of passage 3 umbilical cord mesenchymal stem cells, and Nestin expression was detected in passage 3 cells after 3 days culture in neural induction medium by fluorescence immunochemistry staining. RESULTS AND CONCLUSION:These two methods were both successful to harvest umbilical cord mesenchymal stem cells, but the first adhesion time was earlier in cells cultured by cold trypsin digestion than tissue explant method (P0.05). Under the inverted microscope, cells grew adherently and presented fibroblast-like shape. However, the minority of primary cells under induction of cold trypsin digestion was polygonal, irregular, and had larger cellvolume than those cultured by tissue explant method. Passage 3 cells cultured by tissue explant method showed faster proliferation rate than those cultured by cold trypsin digestion, and at logarithmic growth phase, cells cultured by these two methods were significant different in cellnumber (P<0.05). Two types of cells had a uniform cellphenotype, both of which expressed CD29, CD105, but did not express CD34, CD45. Under induction, passage 3 cells cultured by these two methods were both positive for nestin. These findings indicate that these two methods can both be used to culture umbilical cord mesenchymal stem cells, but the tissue explant method is more suitable for culture of umbilical cord mesenchymal stem cells and exhibits less damage to cells.
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BACKGROUND:There is a high prevalence of spinal tuberculosis in developing countries. Beside systematic chemotherapy, there are stil some patients who need surgical treatment at certain phase of systematic treatment. However, there is no standard method for surgical treatment of spinal tuberculosis yet. OBJECTIVE:To systematical y evaluate the effect of anterior approach and posterior approach through consulting the literatures on the comparison between anterior approach and posterior approach for the treatment of spinal tuberculosis. METHODS:The PubMed database, Medline database, Elseveir database, Wanfang database and CNKI database were searched with the key words of“thoracolumbar tuberculosis, control ed randomized trial, RCT, anterior, posterior”in Chinese and English. The randomized control ed trials on the comparison between anterior approach and posterior approach for the treatment of spinal tuberculosis were included. The operative time, intraoperative blood loss, correction degree of Cobb angle, loss of Cobb angle at final fol ow-up, al ograft fusion time, total hospital stay and the excellent and good rate of the effectiveness were compared by Revman5.1 software. RESULTS AND CONCLUSION:A total of 1 438 articles were screened out, and final y 9 randomized control ed trials were included. The total number of patients was 692, in which 324 were treated with anterior approach and 368 were treated with posterior approach. The operative time of anterior approach of 46.25(40.23, 52.26) minutes was less than that of posterior approach;the intraoperative blood loss of anterior approach of 148.91(135.12, 1 625.70) mL less than that of posterior approach;the correct degree of Cobb angle of anterior approach of 2.40°(2.21°, 4.62°) was smal er than that of posterior approach;the loss of Cobb angle of anterior approach of 0.66°(0.41°, 0.91°) was larger than that of posterior approach;the total hospital stay of anterior approach of 0.34 (-0.32,1.01) days was less than that of posterior approach;the al ograft fusion time of anterior approach was less than that of posterior approach for 0.26 (0.18, 0.34) months;the number of cases with excellent and good effect of anterior approach was more than that of posterior approach for 1.18(-0.48, 2.85);there were significant differences in operative time, intraoperative blood loss, correct degree of Cobb angle, loss of Cobb angle at final fol ow-up and al ograft fusion time between anterior and posterior approaches (P<0.01). The results showed that there were significant differences in operative time, intraoperative blood loss, correct degree of Cobb angle, loss of Cobb angle at final fol ow-up and al ograft fusion time between anterior and posterior approaches for the treatment of spinal tuberculosis, but there were no significant differences in total hospital stay and surgical efficacy.
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BACKGROUND:In order to promote the recovery of nerve function after peripheral nerve injury, we should try to accelerate the regeneration of injured nerve axons on one hand, and try to improve the accuracy of proximal and distal nerve docking on the other hand. OBJECTIVE:To explore the selective regeneration of nerve axons after peripheral nerve injury in models with crush injury and freeze injury. METHODS:Total y 110 healthy male Sprauge-Dawley rats, 8 weeks old, were selected and randomly divided into three groups. The rates received crush injury, freeze injury and normal control ed surgery on femoral nerve trunk. At 2, 3, 6 and 12 weeks after modeling, general behavioral examination was performed, and then pure blue and red fluorescence were used to label the wrong-way ingrowth saphenous nerve and correct ingrowth femoral nerve muscular branches respectively;retrograde labeling the motor neurons in order to observe the distribution and amount of tracer in spinal cord anterior horn;electrophysiological examination was performed at 8 weeks after modeling, and analyzed statistical y. RESULTS AND CONCLUSION:Rats from both crush injury group and freeze injury group displayed decreased motion radius of hind limbs after surgery, and the stretch function was limited;but with time prolonging, the functions were recovered. The motor evoked potentials could be observed at the quadriceps, and there was no significant difference between two groups (P>0.05). The number of neurons stained red in the spinal cord anterior horn was increased gradual y in both crush injury group and the freeze injury group under fluorescence microscope. The number of neurons stained red in the crush injury group was significantly higher than that in the freeze injury group (P<0.05), and the number of blue-stained and purple-stained neurons was decreased gradual y. The results suggest that keeping the perineurium intact can gain accurate docking in axon regeneration and the recovery of injured limb functions, even though the damaged range is large.
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OBJECTIVE:To compare the effects of total knee arthroplasty performed with and without patellar resurfacing. METHODS:All randomized clinical trials relevant to comparison of total knee arthroplasty performed with and without patellar resurfacing between 1966 and 2008 were collected. The quality of the trials was assessed. The outcomes data including the prevalence of reoperations,postoperative anterior knee pain,patient satisfaction,and the improvement in various knee scores were extracted and reviewed systematically. RESULTS:1 731 knees were included in the 15 published trials,including 842 knees in the resurfaced group and 889 in the nonresurfaced group. During the four years following surgery,there was no significant difference between the resurfaced and the nonresurfaced groups in re-operation rate (relative risk=0.56,95% CI 0.17-1.85); whereas the re-operation rate of the nonresurfaced group was significantly greater than resurfaced group 4 years after surgery (relative risk=0.46,95% CI 0.25-0.84). On the prevalence of postoperative anterior knee pain,there was no significant difference between two groups (relative risk=0.65,95% CI 0.35-1.21). No significant difference was found in patient satisfaction (relative risk=1.00,95% CI 0.95-1.05). There was no significant difference between the resurfaced and the nonresurfaced groups in the KSS scores (weighing mean difference:-0.41; 95% CI:-3.08 to 2.26). CONCLUSION:The long-term rate of reoperations in total knee arthroplasty performed with patellar resurfacing was lower than that without patellar resurfacing. There were no significant differences in the terms of prevalence of postoperative anterior knee pain,patient satisfaction,and the improvement in various knee scores between the resurfaced and the nonresurfaced groups. The results of this systematic review should be explained prudently because of some limitations of included trials. For more reliable conclusions,more trials with high quality on methodology are needed.