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Objective:To investigate the therapeutic efficacy of Ilizarov bone shortening-lengthening technique for tibial defects of bone and soft tissue without vascular injury.Methods:A retrospective analysis was made of the 28 patients who had been treated by Ilizarov bone shortening-lengthening technique at Department of Orthopaedics, Wuxi No.9 People's Hospital from January 2007 to October 2017 for tibial de-fects of bone and soft tissue without vascular injury.They were 20 males and 8 females, aged from 18 to 69 years (average, 36.4 years).By the Gustillo classification, 5 cases belonged to type Ⅱ, 6 to type ⅢA and 17 to type ⅢB.Infection was complicated in 17 cases.After debridement or epluchage, the area of skin defects ranged from 4 cm × 3 cm to 16 cm × 5 cm and the length of bone defects from 4.5 to 11.0 cm (average, 6.9 cm).The wound healing, bone healing, functionary recovery of lower extremity and complications were observed postoperatively.Bone healing and functional recovery of lower extremity were evaluated according to the grading of Association for the Study and Application of the Method of Ilizarov (ASAMI).The complications associated with Ilizarov technique were assessed according to the Paley criteria.Results:The follow-up for all the patients lasted from 12 to 45 months (average, 20.5 months).The healing time for wounds ranged from 13 to 35 days (average, 21.9 days), the healing time for lengthened bone from 6 to 12 months (average, 8.9 months), and the healing time for bone defects at the dock sites from 6 to 11 months (8.3 months).According to the ASAMI grading, the bone healing was excellent in 21 cases and good in 7, giving an excellent to good rate of 100%(28/28) while the functionary recovery of lower extremity was excellent in 10 cases, good in 15, fair in 2 and poor in one, giving an excellent to good rate of 89.3%(25/28).The incidence was 14.3%(4/28) for major complications after Ilizarov surgery, 57.1%(16/28) for minor complications, 60.7%(17/28) for overall complications, and 1.7 times for each case.Conclusion:In the treatment of tibial defects of bone and soft tissue without vascular injury, Ilizarov bone shortening-lengthening technique can deal with the difficulties in repair of soft tissue defects, characterized by simplified wound closure, fast and improved bone healing at the dock sites, reduced complications and satisfactory functionary recovery of lower extremity.
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Objective To investigate the regulatory effect of adrenomedullin (AM) on the cell-cell contact formation of podocytes and the possible mechanism.Methods Podocytes were treated with AM (10-7 mol/L),AM combined with a PKA inhibitor H89 (10-4 mol/L),and forskolin (10-5 mol/L) as positive control respectively for 12 hours.Immunofluorescent staining was applied to observe the distribution of cell adhesion molecules and actin-associated proteins.Western blotting assay was used to assess their protein levels.Rho GTPases activity was analyzed by GST-pull down assay and their protein levels were tested by Western blotting.Results AM induced the redistribution of adhesion molecules,actin-associated proteins as well as the F-actin at cell-cell contacts between podocytes.This effect was similar to that of forskolin and could be blocked by H89.The levels of those proteins did not change significantly (P ＞ 0.05).AM up-regulated the activities of RhoA,Rac1 and Cdc42 (P ＜ 0.05),which were partially blocked by H89.The protein levels of Rho GTPases showed no difference compared with the control (P ＞ 0.05).Conclusions AM may promote cell-cell contact formation of podocytes,probably through enhancing the activity of Rho GTPases and then resulting in the redistribution of adhesion molecules,actin-associated proteins and F-actin,which is partially mediated through cAMP-PKA signaling pathway.
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BACKGROUND:With the extensive application of anterior titanium plate, postoperative complications such as dysphagia, titanium loose, screw exit and disc degeneration of neighboring segments induced more and more attention of researchers. However, the application of anterior cervical cage is expected to avoid these complications. ＜br> OBJECTIVE:To observe primary curative effect of anterior cervical cage ROI-C in anterior cervical spine surgery. ＜br> METHODS:A total of 32 patients with cervical spondylosis were treated with anterior cervical cage ROI-C in the Wuxi Ninth Hospital Affiliated to Soochow University from April to December 2013. The cage was implanted to promote interbody fusion. Of 32 cases, 23 cases affected cervical spondylotic myelopathy, 2 cases affected nerve root type cervical spondylosis, 3 cases affected cervical hyperextension injury, 1 case affected cervical disc herniation, 2 cases affected cervical instability and 1 case affected segmental cervical ossification of the posterior longitudinal ligament. Japanese Orthopaedic Association and NDI scores were determined to assess neurological symptoms and functional improvement before internal fixation and during final fol ow-up. Simultaneously, adverse reactions were recorded. ＜br> RESULTS AND CONCLUSION:A total of 32 patients finished the regular fol ow-up for 4 to 8 months. Clinical symptoms and spinal cord function of al patients were obviously improved. No ROI-C loosing or displacement or secondary surgery was found. The average fusion time was 4.2 months (3 to 5 months). Mean score of Japanese Orthopaedic Association was increased from 9.2 points pre-surgery to 13.8 points post-surgery. Japanese Orthopaedic Association and NDI scores were higher during final fol ow-up than before fixation (P＜0.05). These data indicated that ROI-C effectively restored intervertebral height in anterior cervical spine surgery, stably reconstructed cervical vertebra, obtained interbody fusion, effectively avoided related surgical complications induced by plate implantation, improved neurological symptoms and function, and showed good short-term effects.
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BACKGROUND:Previous studies have mainly focused on costal cartilage, articular cartilage, nasal septal cartilage, and auricular cartilage, but in vitro culture of human vertebral endplate cartilage is stil rarely reported. OBJECTIVE: To discuss the feasibility of culture of vertebral endplate chondrocytes from type I neurofibromatosis associated with scoliosis patientsin vitro and to study the biological characters of the chondrocytes. METHODS: Through two-step enzymatic digestion and tissue culture, the chondrocytes from the vertebral endplate of seven type I neurofibromatosis patients isolated and cultured in monolayer and passaged to observe the changes of cel morphology under inverted phase contrast microscope. Colagen type II expression was detected by immunocytochemistry to identify whether the cels had chondrocyte characters. The growth kinetics was detected by using MTT colorimetric assay to draw the growth curve of passage 2 chondrocytes. RESULTS AND CONCLUSION:A few chondrocytes crawled from the cartilage after 2 weeks culture and cels were passaged at 3 weeks. Along with passage going on, the phenotype of chondrocytes was changed from polygonal, round, triangle, and irregular shapes to fusiform. The colagen type II expression in passage 2 cels was positive by immunohistochemical staining. MTT test showed the growth curve of the passage 2 chondrocytes presented a transverse “S”. Cels were found logarithmic growth at days 4-7, reached platform stage at days 8-13, and decreased at day 14. It is an effective and simple procedure by two-step enzymatic digestion and tissue explant method to culture vertebral endplate chondrocytes with high purity and good viability from type I neurofibromatosis patients associated with scoliosisin vitro. Passage 2 chondrocytes from the vertebral endplate exhibit the best viability at days 4-7, which can be used as targets for research of pathogenesis of type I neurofibromatosis with atrophic scoliosis.
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Objective To examine the expression of response gene to complement 32 (RGC-32) in renal tissue of children with IgA nephropathy (IgAN), and to explore its significance. Methods The subjects were 45 children diagnosed as IgAN by renal biopsy. The expression of RGC-32, α-smooth muscle actin (α-SMA) and transforming growth factor β1 (TGF-β1) was examined by immunohistochemistry staining. The correlation of RGC-32 expression with α-SMA,TGF-β1, degree of renal pathological lesions and clinical index in IgAN was assessed by Spearman correlation analysis. Results RGC-32 protein located in renal tubular epithelial cells in normal and IgAN renal tissues. The positive expression index of RGC-32 in nomal group, IgAN mild group, moderate group and severe group was (18.29±6.22)%, (23.90±9.65)%, (31.23±9.86)%,and (34.52±10.63)% respectively. With more severity of renal pathological lesions, the expression of RGC-32 in IgAN was enhanced. The RGC-32 expression was positively correlated with the score of glomerulus and renal interstitium in children with IgAN (r=0.385, 0.347, P＜0.05), as well as α-SMA, TGF-β1 (r=0.594, 0.521, P＜0.01), but was not correlated with Scr, urinary NAG/Cr,Alb/Cr, IgG/Cr, and α1-M/Cr (r =0.117, -0.115, -0.138, -0.176, -0.028, all P ＞0.05).Conclusions RGC-32 protein locates in renal tubular epithelial cells in normal and IgAN renal tissues. RGC-32 may participate in the course of renal tubulointerstitial lesions in children with IgAN, especially in the course of epithelial-mesenchymal transition (EMT) induced by TGF-β1.
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Object: To investigate the effects of Huaiqihuang Granule, a compound Chinese herbal medicine, on expressions of nephrin and podocin of slit diaphragm of glomerular podocytes in rats with adriamycin-induced nephrosis and to explore the mechanism in reducing the proteinuria. Methods: Twenty SD rats were randomly divided into five groups: control group, model group, glucocorticoid group, Huaiqihuang Granule group and Huaiqihuang Granule plus glucocorticoid group. The 24-hour urine was collected 7, 14, 21 and 28 days after adriamycin injection respectively to measure 24-hour urinary protein, and all rats were sacrificed after 28-day treatment. Pathological changes in renal tissues were observed under a light microscope and an electron microscope. Expressions of nephrin and podocin mRNAs in renal cortex were determined by real-time polymerase chain reaction, and protein levels of nephrin and podocin were detected by Western blotting. Results: (1) In the model group and the treatment groups, the level of urinary protein increased significantly from the 14th day. (2) Under the light microscope, inflammatory cells and slight fibroplasia were found in renal interstitium of the model group, but there were less inflammatory cells in renal interstitium in the intervention groups than in the model group. Under the electron microscope, 29 days after adriamycin injection, extensive fusion of foot processes was observed. (3) The expressions of nephrin and podocin were higher in treatment groups than in the model group. (4) Proteinuria level was negatively correlated with the expressions of nephrin mRNA and nephrin and podocin proteins. Conclusion: The above results indicate that Huaiqihuang Granule can maintain the integrity of the slid diaphragram in podocyte, alleviate the lesion of glomerular filtration membrane, and decrease the proteinuria by up-regulating the expressions of nephrin and podocin. Huaiqihuang Granule plus glucocorticoid maybe has better effects than glucocorticoid alone.
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<p><b>OBJECTIVE</b>To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC).</p><p><b>METHODS</b>A monoclonal antibody against AM developed by our laboratory was used to detect the localization of AM protein in rat kidney tissue by avidin-biotin complex immunohistochemistry. The expressions of AM and its receptor CRLR mRNA on cultured glomerular epithelial cells (GEC) and MsC were investigated by Northern blot assay, and the possible effect of AM secreted by GEC on MsC proliferation was observed using [3H]thymidine incorporation as an index.</p><p><b>RESULTS</b>A specific monoclonal antibody against AM was succesfully developed. AM was immunohistochemically localized mainly in glomeruli (GEC and endothelial cells), some cortical proximal tubules, medullary collecting duct cells, interstitial cells, vascular smooth muscle cells and endothelial cells. Northern blot assay showed that AM mRNA was expressed only on cultured GEC, but not on MsC, however, AM receptor CRLR mRNA was only expressed on MsC. GEC conditioned medium containing AM can inhibit MsC growth and AM receptor blocker CGRP8-37 may partially decreased this inhibitory effect.</p><p><b>CONCLUSION</b>AM produced by GEC inhibits the proliferation of MsC, which suggests that AM as an important regulator is involved in glomerular normal physiological functions and pathologic processes.</p>