Objective To evaluate the effects of two different epidermal growth factor receptor tyrosine kinase inhibitors ( EGFR-TKIs) , Gefitinib and Erlotinib, on lung fibrosis induced by bleomycin.Methods Forty BALB/c female mice were randomly divided into four groups, ie. a control group( saline given orally and intratracheally) , a fibrosis group( saline given orally with bleomycin instillation) , a Gefitnib group( Gefitnib 20 mg/kg given orally with bleomycin instillation) , and an Erlotinib group ( Erlotinib25 mg/kg given orally with bleomycin instillation) . Bleomycin ( 3 mg/kg) was intratracheally instilled on the first day. Gefitinib or Erlotinib was given orally daily and normal saline as control. Then they were sacrificed by abdominal aortic bleeding 14 days after the bleomycin instillation. The left lung was stained with HE and Masson’s trichrome staining respectively for pathological examination. Total EGFR and phosphorylated EGFR were detected by immunohistochemistry. Hydroxyproline ( HYP) assay was performed in the right lung.Results Both Gefitinib and Erlotinib significantly reduced lung collagen accumulation and the content of HYP. Immunohistochemistry revealed that phosphorylation of EGFR in lung mesenchymal cells induced by bleomycin was inhibited. Furthermore, there was no difference between Gefitinib and Erlotinib in inhibiting lung fibrosis. Conclusion Our findings suggest that, in the preclinical setting, EGFR-TKIs may have aprotective effect on lung fibrosis induced by bleomycin.
Objective To investigate the role of alveolar macrophages ( AMs ) in airway inflammation of smoke-induced COPD rat model and its possible regulating mechanism. Methods Twelve Wistar rats were randomly divided into a COPD group and a control group. The rat model of COPD was established with smoke exposure and LPS intrathacheal instillation. Bronchoalveolar lavage fluid ( BALF)was collected for measurement of total and differential cell counts. Then AMs were isolated and identified byimmunofluorescence. Western blot was employed to analyze the cytoplasmic and nuclear NF-κB p65 expression of AMs. The concentrations of TNF-α,macrophage inflammatory protein 2 ( MIP-2) and IL-10 in cell culture supernatantwere assayed by ELISA.Results The scores of bronchitis and mean liner intercepts in the COPD group were significantly higher than those in the control group [ 4. 33 ±1. 16 vs. 1. 33 ±0. 58,P =0. 016; ( 168. 77 ±11. 35) μm vs. ( 93. 61 ±4. 16) μm, P = 0. 000) ] . The total cell count in BALF of the COPD group was significantly higher than that in the control group ( P lt; 0. 05) , and the AMs and neutrophils were predominant [ ( 72. 00 ±2. 22) % and ( 18. 29 ±8. 34) % ] . The cytoplasmic NF-κB p65 expression of AMs in the COPD group was significantly lower , while the nuclear NF-κB p65 expression was significantly higher ( P lt; 0. 05) compared with the control group. The ELISA results showed that the concentrations of TNF-αand MIP-2 in culture supernatant of AMs in the COPD group were significantly higher than those in the control group ( P lt;0. 05) , while the concentration of IL-10 was not significantly different between the two groups ( P gt;0. 05) . Conclusions COPD rat model was established successfully with smoke exposure and LPS intratracheal instillation with a profile of macrophage-based chronic inflammation and increased secretion of TNF-αand MIP-2. The mechanismis closely related to activation of NF-κB.
ObjectiveTo compare two different ways to establish mouse model with acute lung injury (ALI) via intratracheal instillation or intraperitoneal injection of lipopolysaccharide (LPS). MethodsBALB/c mice received intraperitoneal/intratracheal administration of LPS or sham operation. Wet/dry lung weight ratio, protein concentration in bronchoalveolar lavage fluid (BALF), and lung tissue histology were examined at 0, 1, 2, 6, 12, 18, 24, 48 h after LPS administration. Tumor necrosis factor-α (TNF-α) in BALF and serum was assayed with ELISA method. ResultsLPS treatment significantly increased wet/dry lung weight ratio, BALF protein concentration and TNF-α concentration in serum and BALF. Lung tissue was damaged after LPS challenge. The mice received LPS intraperitoneal injection got a more significant lung edema than those received LPS intratracheal instillation. Inversely, LPS intratracheal instillation induced more severed microstructure destruction. ConclusionsALI animal model by LPS intratracheal instillation or intraperitoneal injection induces inflammation and tissue damage in lung. However, the degree of tissue damage or self-healing induced by two methods is different. Therefore the decision of which way to establish ALI model will depend on the study purpose.
Objective To explore the effectiveness of staged treatment of open Pilon fracture combined with soft tissue defect. Methods Between June 2007 and December 2012, 18 cases of open Pilon fracture combined with soft tissue defect were treated. There were 14 males and 4 females with an average age of 35 years (range, 19-55 years). The causes of injury included falling from height in 12 cases, traffic accident in 4 cases, and crushing by machine in 2 cases. According to AO classification, 1 case was classified as type B2 fracture, 3 cases as type B3 fracture, 5 cases as type C1 fracture, 5 cases as type C2 fracture, and 4 cases as type C3 fracture. Sixteen cases accompanied by fibular fracture (14 cases of simple fibular fracture and 2 cases of communicated fibular fracture). According to Gustilo classification, the soft tissue injuries were all type IIIB. In first stage, debridement and vaccum sealing drainage combined with external fixation were performed; open reduction and internal fixation of simple fibular fracture were used. In second stage, open reduction and internal fixation of Pilon fracture and communicated fibular fracture were performed, and the flaps of 6 cm × 5 cm to 18 cm × 14 cm were applied to repair soft tissue defect at the same time. The donor site was repaired by skin graft. Results Partial necrosis occurred in 2 flaps, the other 16 flaps survived completely. The incisions of donor sites healed by first intention, the skin graft survived completely. The average follow-up interval was 12 months (range, 6-24 months). The X-ray films showed that the bone healing time ranged from 5 to 8 months (mean, 6 months). No internal fixation failure was found. At last follow-up, the average range of motion of the ankle joint was 37° (range, 26-57°). According to the American Orthopedic Foot and Ankle Society (AOFAS) scale, the average score was 80.2 (range, 72-86). Traumatic arthritis occurred in 2 cases (11%). Conclusion The staged treatment has the advantages of accurate evaluation of soft tissue injury, shortened cure time, good reduction of the articular surface, and reduced incidence of infection, so it is an optimal method to treat open Pilon fracture combined with soft tissue defect.
ObjectiveTo explore the clinical application and effectiveness of antibiotic-loaded cement spacer combined with free fibular graft in the staged treatment of infectious long bone defect in the lower extremity. MethodsA retrospective analysis was made on the clinical data from 12 patients with infectious long bone defect in the lower extremity between June 2010 and June 2012. Of the 12 cases, there were 9 males and 3 females with an average age of 33 years (range, 19-46 years), including 3 cases of femoral shaft bone defect, 7 cases of tibial shaft bone defect, and 2 cases of metatarsal bone defect. The causes were traffic accident injury in 7 cases, crashing injury in 3 cases, and machine extrusion injury in 2 cases. The length of bone defect ranged from 6 to 14 cm (mean, 8 cm). The soft tissue defect area ranged from 5.0 cm×3.0 cm to 8.0 cm×4.0 cm companied with tibial shaft and metatarsal bone defect in 9 cases. The sinus formed in 3 femoral shaft bone defects. The time between injury and operation was 1-4 months (mean, 2 months). At first stage, antibiotic-loaded cement spacer was placed in the bone defect after debridement and the flaps were used to repair soft tissue defect in 9 cases; at second stage (6 weeks after the first stage), defect was repaired with free fibular graft (7-22 cm in length, 14 cm on average) after antibiotic-loaded cement spacer removal. The area of the cutaneous fibular flap ranged from 6.0 cm×4.0 cm to 10.0 cm×5.0 cm in 10 cases. ResultsAll wounds healed by first intention, and the healing time was 12-18 days, 14 days on average. Twelve cases were followed up 12-36 months (mean, 17 months). Bone healing time ranged from 4 to 6 months (5.5 months on average). The cutaneous fibular flap had good appearance. The function at donor site was satisfactory; no dysfunction of the ankle joint or tibial stress fracture occurred after operation. The mean Enneking score was 25 (range, 20-28) at last follow-up. ConclusionInfection can be well controlled with the antibiotic-loaded cement spacer during first stage operation, and free fibular graft can increase the bone defect healing rate at second stage. Staged treatment is an optimal choice to treat infectious long bone defect in the lower extremity.