OBJECTIVE To investigate the effect of allogeneic decalcified bone graft in the treatment of nonunion in children. METHODS From April 1990 to September 1997, 17 children with nonunion were adopted in this study. Among them, there were 10 boys and 7 girls, the age ranged from 2 to 13 years. The allogeneic decalcified bone graft taken from fresh corpse by aseptic manipulation were used to repair 3 cases of congenital nonunion and 14 cases of acquired nonunion. RESULTS All cases were followed up 2 to 9 years after operation, 9 cases were cured successfully and 7 cases were cured after twice operations. One case of congenital pseudoarthrosis of the tibia was operated twice and there were callus grown half years after the second operation, but reoccurred after one year. Although there were 1.5 cm to 3 cm shortening deformity of extremity including another 2 cases of congenital pseudoarthrosis of the tibia and 5 cases of nonunion caused by chronic osteomyelitis, the function of joint was normal. CONCLUSION Treatment of nonunion in children by allogeneic decalcified bone graft is a valuable technique in clinical practice. It is characterised by high capacity of osteoinduction, low antigenicity, rapid bony union, plentiful source of bone graft and convenient utilization.
Objective To study the effect on expression of high mobility group box-1 （HMGB1） mRNA for the expression of zonula occludens-1 （ZO-1） in ileum tissues， and to explore the possible mechanism of intestinal mucosal barrier injury in rats with acute necrotizing pancreatitis （ANP）. Methods Ninety-six male Wistar rats were divided randomly （random number method） into ANP group， ethyl pyruvate （EP）group， and sham operation group. Eight rats of 3 groups were killed to get abdominal aortic blood and ileal tissues at 6， 12， 24， and 48h after operation， respectively.The levels of plasma amylase （AMY） ， D-lactate acid， and the activity of malonyl dialdehyde （MDA） in the ileum tissues were determined by using automatic biochemical analyzer， improved enzymatic spectrophotometry， and thiobarbituric acid （TAB） colorimetry respectively. The pathological changes of ileum tissues were observed under microscopy by HE staining， the expression of ZO-1 protein in ileum tissues was observed by immunohistochemistry （SP method）， and the expressions of HMGB1 mRNA and ZO-1 mRNA in ileum tissues were detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results Compared with ANP group at the same time， levels of AMY， D-lactate acid， and MDA in ileum tissues of EP group were all significantly lower （P＜0.05）. The expression level of HMGB1 mRNA increased at 6 h while ZO-1 mRNA decreased in ANP group. Compared with ANP group at the same time， the expression level of HMGB1 mRNA of EP group was significantly lower while ZO-1 mRNA was higher （P＜0.05）， and the pathological damage in ileum tissues was lighter. Conclusions The decreased expression of ZO-1 in ileum tissues is one of the vitalcauses for intestinal mucosal barrier injury in ANP， and it probably occurs in case of the excessive expression of HMGB1.
ObjectiveTo study the effect and mechanism of lipopolysaccharide (LPS) on osteoclasts formation and its bone resorption function.MethodsBone marrow-derived macrophages (BMMs) were extracted from the marrow of femur and tibia of 4-week-old male C57BL/6 mice. Flow cytometry was used to detect BMMs. The effect of different concentrations of LPS (0, 100, 200, 500, 1 000, 2 000 ng/mL) on BMMs activity was examined by cell counting kit 8 (CCK-8) activity test. In order to investigate the effect of LPS on osteoclastogenesis, BMMs were divided into macrophage colony-stimulating factor (M-CSF) group, M-CSF+receptor activator of nuclear factor κB ligand (RANKL) group, M-CSF+RANKL+50 ng/mL LPS group, M-CSF+RANKL+100 ng/mL LPS group. After the completion of culture, tartrate resistant acid phosphatase (TRAP) staining was used to observe the formation of osteoclasts. In order to investigate the effect of LPS on the expression of Connexin43, BMMs were divided into the control group (M-CSF+RANKL) and the LPS group (M-CSF+RANKL+100 ng/mL LPS); and the control group (M-CSF+RANKL), 50 ng/mL LPS group (M-CSF+RANKL+50 ng/mL LPS), and 100 ng/mL LPS group (M-CSF+RANKL+100 ng/mL LPS). The expressions of Connexin43 mRNA and protein were detected by Western blot and real-time fluorescent quantitative PCR, respectively. In order to investigate the effect of LPS on osteoclast bone resorption, BMMs were divided into M-CSF group, M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group. Bone absorption test was used to detect the ratio of bone resorption area.ResultsThe flow cytometry test confirmed that the cultured cells were BMMs, and CCK-8 activity test proved that the 100 ng/mL LPS could promote the proliferation of BMMs, showing significant differences when compared with the 0, 200, 500, 1 000, and 2 000 ng/mL LPS (P<0.05). TRAP staining showed no osteoclast formation in M-CSF group. Compared with M-CSF+RANKL group, the osteoclasts in M-CSF+RANKL+50 ng/mL LPS group and M-CSF+RANKL+100 ng/mL LPS group were larger with more nuclei, while the osteoclasts in M-CSF+RANKL+100 ng/mL LPS group were more obvious, and the differences in the ratio of osteoclast area between groups were statistically significant (P<0.05). Western blot result showed that the relative expression of Connexin43 protein in LPS group was significantly higher than that in control group (P<0.05). Real-time fluorescent quantitative PCR showed that the relative expression of Connexin43 mRNA in control group, 50 ng/mL LPS group, and 100 ng/mL LPS group increased gradually, and the differences between groups were statistically significant (P<0.05). Bone resorption test showed that osteoclast bone resorption did not form in M-CSF group, but the ratio of bone resorption area increased gradually in M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group, and the differences between groups were statistically significant (P<0.05).ConclusionLPS at concentration of 100 ng/mL can promote the expression of Connexin43, resulting in increased osteoclastogenesis and enhanced osteoclastic bone resorption.
ObjectiveTo study the expression of zonula occluden-1 (ZO-1) in ileum tissues and the possible mechanism of intestinal mucosal barrier injury in rats with severe acute pancreatitis (SAP). MethodsFifty SD male rats were randomly divided into sham operation group and SAP group, then SAP group was divided into four subgroups with 10 rats in each subgroup according to the sampling time of 3, 6, 12, and 24 h. The SAP model was made by injecting 5% bovine sodium deoxycholate into biliarypancreatic duct with Aho’s method. The rats were killed at 3, 6, 12, and 24 h after making model. The rats in the sham operation group were killed directly. Tumor necrosis factor-α (TNF-α), diamine oxidase (DAO), and histological changes in pancreatic and intestinal pathologies were observed. At the same time, the ZO-1 protein and mRNA expressions of ileum tissues were detected by immunohistochemistry and RT-PCR, respectively. ResultsCompared with the sham operation group 〔TNF-α: (10.83±0.96) ng/L; DAO: (354.79±3.67) U/L; ZO-1 protein: (10.40±0.45) score; ZO-1 mRNA: 0.878±0.014 8〕, the levels of TNF-α at different time 〔3 h: (125.30±0.94) ng/L; 6 h: (181.89±4.93) ng/L; 12 h: (230.58±1.28) ng/L; 24 h: (198.89±4.83) ng/L〕 were significantly higher (Plt;0.05), the activities of DAO 〔3 h: (235.77±0.67) U/L; 6 h: (117.22±5.58) U/L; 12 h: (106.69±1.39) U/L; 24 h: (91.18±1.09) U/L〕 were significantly lower (Plt;0.05), ZO-1 protein 〔3 h: (8.70±0.22) score; 6 h: (3.73±0.19) score; 12 h: (3.92±0.22) score; 24 h: (4.29±0.30) score〕 and mRNA (3 h: 0.806±0.020 7; 6 h: 0.370±0.015 8; 12 h: 0.502±0.019 2; 24 h: 0.562±0.030 3) expressions of the ileum tissues were significantly lower (Plt;0.05) in the SAP group; Meanwhile, the necrosis of ileum mucous membrane chorioepithelium, angiorrhexis and hemorrhage, and inflammatory cell infiltration in the pancreatic and ileum tissues were also observed. ConclusionThe decrease of expression of ZO-1 in ileum tissues is one of the vital causes for mucosal barrier injury in SAP, probably through acts the excessive release of inflammatory cytokines TNF-α and the decrease of DAO activity.
Objective To investigate a change in the differentiation and biological function of the cultured rat fibroblast (FB) transfected by the myoblast determining gene (MyoD) and the connexin 43 (Cx43) gene and to explore the possible mechanism of the MyoD and Cx43 genes on treatment of ischemic heart disease (IHD). Methods The gene cloning technology was used to construct the eukaryotic expressed plasmid vector pLenti6/V5-DEST-MyoD and pLenti6/V5DEST-Cx43 in which MyoD cDNA or Cx43 cDNA was inserted. The RFL-6 FB cells were transfected with exogenetic MyoD cDNA or Cx43 cDNA via lipofectamine, followed by the Blasticidin (50 μg/ml) selection, according to the lentiviral expression system (ViraPower) protocol. The expression and the biological functions of MyoD and Cx43 in the transfectants were testified by RT-PCR, Western blot, and molecular and immunocytochemical methods. The mophological structure changes of the cells were observed under microscope before and after the transfection. Results The expression of MyoD and Cx43 was detected in the MyoD and Cx43 genes transfected FB with RT-PCR and Western blot. The immunocytochemical methods indicated the expressionsof the MyoD and Cx43 genes, while desmin and αactin were found in these cells. The myotubes were found from the cultures incubated a week in the differentiation medium, in which the transfected cells had a characteristic of the filamentsin their cytoplasm and showed a myoblast morphology. Conclusion MyoD cDNA can induce the cultured FB to differentiate into the myoblasts and Cx43 cDNA can enhance the gap junctional intercellular communication between the cell and the cell. Thus, a further experimental foundation for the therapy of IHD can be provided.
ObjectiveTo investigate the prevalence, severity and consequences of acute kidney injury (AKI) in the patients who underwent total cavopulmonary connection (TCPC).MethodsThe clinical data of TCPC patients in our center from January 1, 2010 to December 31, 2014 were collected and retrospectively analyzed. The patients with renal replacement therapy, missing serum creatinine data before operation or combined with valve procedures were excluded. We identified whether AKI was associated with hospital length of stay, ICU duration, mechanical ventilation duration, hospital acquired infection, and early mortality by univariable and multivariable analyses.ResultsA total of 163 patients were included. AKI occurred in 57% of patients (n=93), mild AKI in 26.4% (n=43), moderate AKI in 12.3% (n=20) and severe AKI in 18.4% (n=30). Compared with the no AKI group, the AKI group had higher hospital acquired infection rate (15.1% vs. 0.0%, P<0.001). AKI was independently associated with hospital length of stay (median, 10 d, 95%CI 3.9-16.0, P=0.001), ICU duration (median, 103.9, 95%CI 48.6-159.2, P<0.001) , but not associated with mechanical ventilation duration (median, 8 h vs. 7 h, P=0.529).ConclusionPostoperative AKI in the patients undergoing TCPC is common. AKI is associated with higher hospital acquired infection rate, longer hospital length of stay and ICU duration, but not associated with mechanical ventilation duration.
目的 总结反复黏膜感染的IgA肾病的临床病理特点。 方法 采用单中心流行病学调查及回顾性研究，收集2006年1月－2009年12月253例经肾活检确诊为IgA肾病的住院患者的临床病理资料，对114例反复黏膜感染的IgA肾病患者（A组）及139例偶发或从未发生黏膜感染的IgA肾病患者（B组）的临床病理指标进行比较。 结果 A组患者年龄较B组小（t=2.913，P=0.004），临床表现无明显症状者比例较B组多，临床分型以反复发作肉眼血尿型为多，病理分级较B组轻（Z=?2.042，P=0.041），IgA+IgM+C3沉积率高（P＜0.001）；B组IgA+IgG+C3沉积率高（P＜0.001），纤维连接蛋白沉积率高（P＜0.001）。 结论 反复黏膜感染的IgA肾病组患者年龄小，反复发作肉眼血尿型多，临床表现及病理分级较轻；两组FN沉积及免疫球蛋白沉积的主要类型不同，提示两组发病机制可能有所不同。