Angiotensin II activates connective tissue growth factor and induces extracellular matrix changes involving Smad/activation and p38 mitogen-activated protein kinase signalling pathways in human dermal fibroblasts.

Angiotensin II (Ang II) stimulation has been shown to regulate proliferation of skin fibroblasts and the production of extracellular matrix, which are very important processes in skin wound healing and fibrosis; however, there is little knowledge about the mechanisms involved in this process. We investigated the molecular aspects of this system with regards to Ang II in human dermal fibroblasts (HDF) and its potential role in fibrosis. Fibroblasts derived from human skin were subjected to examine differential relative gene and protein expression after transfection with specific reporter expression vectors and Ang II in vitro. In growth-arrested HDFs, Ang II treatment for 20 min caused acute activation of Smad2 phosphorylation, Smad overexpression and Smad-dependent gene transcription. The angiotensin type 1 (AT1) antagonist losartan diminished Ang II-induced Smad activation. The blockade of endogenous transforming growth factor-beta1 did modify the activation of Smad caused by Ang II. The p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 diminished Ang II-induced Smad2 phosphorylation. Transient transfection with Smad7, which interferes with receptor-mediated activation of Smad2, diminished Ang II-induced connective tissue growth factor promoter activation, gene and protein expression and fibronectin, type I procollagen and type III procollagen overexpression, showing that Smad activation is involved in Ang II-induced dermal fibrosis. Our results show that Ang II activation of Smad2 occurs via the AT1 receptor, but not the AT2 receptor. Activation of Smad2 required p38 MAPK but not p42/p44 MAPK or the epidermal growth factor receptor.

Adenotonsillectomy outcomes regarding bone age and osteocalcin in treatment of obstructive sleep apnea syndrome in children.

BACKGROUND: To investigate the effect of adenotonsillectomy (AT) on bone development, quality of life and polysomnography evaluation in children with obstructive sleep apnea syndrome (OSA). METHODS: Preoperative and postoperative (6 months) physical examination, PSG, bone age (BA) and osteocalcin (OC) evaluation were performed on the selected OSA children (n=92) and the healthy children (n=87). The OSA children were also scored based on the OSA 18-item questionnaire. A two-year follow-up was conducted to evaluate BA and OC changes. RESULTS: After AT, 81 (88.04%) OSA children recovered completely, eight (8.70%) achieved remarkable improvements, and three (3.26%) achieved moderate improvements. In the OSA children, postoperative OSA 18-item score and the scores of the five domains were significantly higher than preoperative ones. Compared with the preoperative, body mass index (BMI), weight for age Z-sores, height for age Z-sores, weight for height Z-sores and BMI Z-score in the OSA group 6 months after the operation were significantly increased, but no significant difference was detected between the OSA and the control group. The changes of BA and chronological age in the OSA group were significantly different from those in the control group. Two years after AT, BA between the two groups was no longer significantly different. Preoperative serum OC in the OSA group was lower than that in the control group, but increased to normal levels 6 months after AT. Correlation analysis showed serum OC levels were negatively correlated with apnea hyponea index, obstructive apnea index, arousal index, and lowest oxygen saturation. CONCLUSIONS: After AT, bone growth and development in children with OSA recovered gradually, and the serum OC levels decreased to the normal level. Therefore, preventive measures and positive treatments should be applied to minimize the negative effects of OSA in children.

Clinical significance of FABP2 expression in newborns with necrotizing enterocolitis.

BACKGROUND: This meta-analysis aimed to determine the role of human fatty acid binding protein 2 (FABP2) expression in the diagnosis of necrotizing enterocolitis (NEC) of newborns. DATA SOURCES: Eligible studies for further statistical analysis were identified from various databases including PubMed, Expert Medica Database, Web of Science, Cochrane Library, Google Scholar, China BioMedicine and China National Knowledge Infrastructure. Random effects model was used, and summary standardized mean difference (SMD) with its 95% confidence interval (CI) was calculated to assess the association of FABP2 expression and NEC. RESULTS: Ten articles which included 572 infants (262 infants with NEC and 310 healthy controls) were included in the current meta-analysis. FABP2 showed a positive relationship with NEC of newborns (SMD=2.88, 95% CI=2.09-3.67, P<0.001). And FABP2 expression was higher in patients with advanced stage of NEC (stage III or stage II+III) than in those with early stage of NEC (stage I) (SMD=-0.48, 95% CI=-0.87 to -0.09, P=0.015). Ethnicity-stratified analysis yielded significantly different estimates with a high FABP2 expression in NEC in both Caucasians (SMD=3.16, 95% CI=1.90-4.43, P<0.001) and Asians (SMD=2.57, 95% CI=1.50-3.64, P<0.001). Sample-based subgroup analysis showed that FABP2 expression was positively correlated with neonatal NEC in both urinary- and blood-sample subgroups (all P<0.05). CONCLUSION: The results prove that the high FABP2 expression is related to the damage to intestinal cells, which may be a possible early detection marker identifying neonatal NEC.

[Predictive value of radiological parameter for re-displacement of conservative treatment of distal radius fracture].

OBJECTIVE: To analyze the radiological parameters of the conservatively in treating distal radius fracture and investigate whether the final re-displacement can be predicted after primary reduction. METHODS: From January 2013 to June 2014,212 patients with distal radial fracture conservatively were treated in our hospital, 107 patients of them were excluded because of their incomplete radiological parameters;the remaining 105 patients were available for radiological were assessed after injury, there were 56 male patients and 49 female patients in this study, the average age of the patients was 51 years old (ranged from 22 to 80 years). According to AO classification, there were 47 cases of type A2 and C1, and 58 cases of type A3, C2, C3. All patients were treated by closed reduction and below-elbow cast immobilization for 4 to 6 weeks. All patients were followed up for 3 to 6 months (means 4.5 months) by X-ray, all fractures were healed. Standard AP and lateral radiographic examination was conducted before reduction and after reduction and bony consolidation,the dorsal angulation and the radial angle were measured at each time point. The linear regression was used for the analysis to find out whether the final re-displacement can be predicted after primary reduction. RESULTS: Among 105 patients,the significant correlations were found for the dorsal angulation between the reduction time and the end time (r = 0.82) and for the radial angulation between the reduction time and end time (r = 0.85). CONCLUSION: The dorsal angulation and the radial angulation after complete healing can be predicted from linear the regression functions. Due to the possibility of predicting the end result, whether the fracture should receive further conservative treatment or surgical treatment can be decided immediately.

T Lymphocyte Subsets and Cytokines in Rats Transplanted with Adipose-Derived Mesenchymal Stem Cells and Acellular Nerve for Repairing the Nerve Defects.

OBJECTIVE: The aim of this study was to explore the immunity in rats transplanted with adipose-derived mesenchymal stem cells (ADSCs) and acellular nerve (ACN) for repairing sciatic nerve defects. METHODS: ADSCs were isolated from the adipose tissues of Wistar rats. Sprague-Dawley rats were used to establish a sciatic nerve defect model and then divided into four groups, according to the following methods : Group A, allogenic nerve graft; Group B, allograft with ACN; Group C, allograft ADSCs+ACN, and Group D, nerve autograft. RESULTS: At the day before transplantation and 3, 7, 14, and 28 days after transplantation, orbital venous blood of the Sprague-Dawley rats in each group was collected to detect the proportion of CD3(+), CD4(+), and CD8(+) subsets using flow cytometry and to determine the serum concentration of interleukin-2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) using enzyme-linked immunosorbent assay (ELISA). At each postoperative time point, the proportion of CD3(+), CD4(+), and CD8(+) subsets and the serum concentration of IL-2, TNF-α, and IFN-γ in group C were all near to those in group B and group D, in which no statistically significant difference was observed. As compared with group A, the proportion of CD3(+), CD4(+), and CD8(+) subsets and the serum concentration of IL-2, TNF-α, and IFN-γ were significantly reduced in group C (p<0.05). CONCLUSION: The artificial nerve established with ADSCs and ACN has no obvious allograft rejection for repairing rat nerve defects.

Serum Brain-derived neurotrophic factor levels in Chinese children with autism spectrum disorders: a pilot study.

Brain-derived neurotrophic factor (BDNF) plays a critical role in the pathogenesis of Autism spectrum disorders (ASD). The purpose of this study was to investigate the potential role of BDNF in Chinese children with ASD. Sixty patients (48 male, 12 female) diagnosed with ASD and 60 healthy sex and age control subjects were assessed for serum BDNF content at admission. BDNF were assayed with enzyme-linked immunosorbent assay methods, and severity of ASD was evaluated with the Childhood Autism Rating Scale (CARS) Score. The results indicated that the median serum BDNF levels were significantly (P<0.0001) higher in children with ASD as compared to normal cases [17.6(IQR: 13.7-21.4) ng/ml and 11.5(9.6-13.8) ng/ml, respectively]. Based on the receiver operating characteristic (ROC) curve, the optimal cut-off value of serum BDNF levels as an indicator for auxiliary diagnosis of autism was projected to be 15.0 ng/ml. Further, we found that an increased risk of ASD was associated with BDNF levels >15.0 ng/ml (adjusted OR 10.4, 95% CI: 4.39-29.32) after adjusting for above possible confounders. Our study demonstrated that serum BDNF levels were associated with ASD, and higher levels could be considered as an independent risk factor of ASD.

[Characteristics and replantation of degloving injury of distal finger].

OBJECTIVE: To explore clinical characteristics and replantation methods of degloving injury of distal finger. METHODS: From 2004 to 2009,18 cases of 19 distal finger degloving were admitted, and included 14 males and 4 femals with an average age of 31 years old ranging from 18 to 51 years old. The distal finger degloving injury was divided into 3 types according to the different levels of degloveing digital artery and skin involving 6 fingers of type I, 10 fingers of type II, 3 fingers of type III. Among them, 3 cases of 4 fingers were failed to be replantaed due to severed injured digital artery, and 15 cases of 15 distal finger degloving injury were replanted with microsurgical technique. RESULTS: Among 15 patients (15 fingers) conpleted the reimplant operation, 13 fingers were survived, 2 fingers were necrosis after operation. Thirteen survived fingers were followed up from 6 to 24 months (averaged 14 months). The appearance of injured fingers and nails obtained satisfactory results. According to Chinese Hand Surgery Society Criteria for function assessment replantation, the results were excellent in 9 cases, good in 3 cases and poor in 1. CONCLUSION: Replantation of distal degloving injury is effective and it should strive for replantation.

[Clinical significance of MESS scoring system in the treatment of fractures of lower limb combined with vascular injuries].

OBJECTIVE: To study the clinical significance of MESS scoring system in the treatment of fractures of lower limb combined with vascular injuries, and to evaluate its reliance. METHODS: From March 2006 to March 2008, 28 patients with fractures of lower limb combined with vascular injuries were graded by MESS scoring system. There were 17 patients were male and 11 patients were female, ranging in age from 23 to 53 years, averaged 38 years. Seventeen patients had fractures at the superior segment of tibia and fibia, 7 patients had fractures at the inferior segment of femur, and other 4 patients had dislocation of knee joint. Among the patients, 18 patients had MESS scores more than 7.0 point, in which 13 patients were treated with one-stage amputation, 5 patients were treated with two-stage amputation; the other 10 patients had the MESS scores less than 7.0 point, and were treated with open reduction and internal fixation, in which 8 patients were treated with transplantation of great saphenous vein to repair blood vessles, and 2 patients were treated with vascular end to end anastomosis. RESULTS: Among the patients, including 18 patients whose MESS scores more than 7.0 point were treated with one-stage or two-stage amputation, and 10 patients whose MESS scores less than 7.0 point were treated with limb salvage operations, all the limbs survived. During the follow-up period (ranged from 0.5 to 1 year, the movement and sensory function of the limbs recovered well. CONCLUSION: MESS is a simple and reliable tool to determine the proper strategy for the patients suffering from vascular injuries with fractures.

Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist inhibits transforming growth factor-beta1 and matrix production in human dermal fibroblasts.

Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonists are increasingly used in patients with diabetes, and some studies have suggested a beneficial effect on organ fibrosis, but their effects on dermal cell growth and extracellular matrix (ECM) turnover are unknown. To investigate the effect of the PPAR-gamma agonist troglitazone on cell growth and matrix production in human dermal fibroblasts (HDF), HDF were cultured and grown in a different concentration of troglitazone. PPAR-gamma expression and matrix production were measured in HDF in the presence of troglitazone. The mRNA expressions of TGF-beta1, collagen I (Col I) and fibronectin (FN) were determined by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). The protein of transforming growth factor-beta1 (TGF-beta1) was determined by enzyme-linked immunosorbent assay (ELISA) and proteins of Col I and FN were determined by Western blotting. The mRNA expression of TGF-beta1, Col I and FN were significantly decreased in HDF in 15-30 micromol l(-1) troglitazone compared to the control group with Dulbecco's modified Eagle's medium (P<0.01). An obvious decrease of TGF-beta1 protein was found in troglitazone-treated groups as compared to the control group (P<0.05). Exposure of HDF to troglitazone reduced col I secretion (P<0.05), and fibronectin secretion (P<0.05). This study suggests that PPAR-gamma agonist will provide a novel approach with therapeutic potential in dermal fibrosis, such as hypertrophic scar, keloid and so on.

Contribution of epidermal stem cells to hypertrophic scars pathogenesis.

Hypertrophic scars are fibroproliferative disorders of excessive wound healing due to an imbalance between synthesis and degradation and the mechanism leading to hypertrophic scars formation is poorly understood and currently no successful treatment modality exists. We hypothesize epidermal stem cells (ESCs), which could inhibit epidermal fibrosis, plays a substantial contributory role in the pathogenesis of hypertrophic scars. Accepting the hypothesis to be correct, a therapy that inhibits cell and extracellular matrix proliferation can be used to prevent the hypertrophic scars formation. Current therapies are only partially effective and safe because they couldn't inhibit the cell and extracellular matrix proliferation and eliminate other relative factors of hypertrophic scars formation at all, such as: absence of epidermal-mesenchymal interaction, and at the same time inducing death (apoptosis and necrosis) of other normal cells. A more efficient prevention of hypertrophic scars could be achieved using tissue engineering skin enriched with ESCs and introduced recombinant genes into ESCs which could inhibit hypertrophic scars formation.

Activation of peroxisome proliferator-activated receptor-gamma inhibits transforming growth factor-beta1 induction of connective tissue growth factor and extracellular matrix in hypertrophic scar fibroblasts in vitro.

Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligands have been recently reported to have beneficial effects on organ fibrosis. However, their effects on extracellular matrix (ECM) turnover in hypertrophic scar fibroblasts (HSFs), and the related molecular mechanisms are unknown. HSFs were cultured and exposed to different concentration PPAR-gamma ligands in the presence of transforming growth factor-beta1 (TGF-beta1). In growth-arrested HSFs, a PPAR-gamma natural ligand (15-deoxy-D12,14-prostaglandin J2, 15d-PGJ2) and a synthetic ligand (GW7845) dose-dependently attenuated TGFbeta1-induced expression of Connective tissue growth factor (CTGF), collagens and fibronectin. Furthermore, the suppression of CTGF mRNA and protein expression are relieved by pretreatment with an antagonist of PPAR-gamma (GW9662). Moreover, GW7845 and 15d-PGJ2 partially inhibited the expression and phosphorylation of the TGF-beta1/Smad pathway. These results suggest that in TGFbeta1-stimulated HSFs, PPAR-gamma ligands caused an antiproliferative effect and reduced ECM production through mechanisms that included reducing CTGF expression, and a crosstalk between PPAR-gamma and Smad may be involved in the inhibitory effects of PPAR-gamma ligands.