[Therapeutic effect of human umbilical cord mesenchymal stem cells on airway remodeling in an asthma model of rat and its possible mechanism].

Objective: To observe the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on airway remodeling in asthma model of rat and its possible mechanism. Methods: hUC-MSCs were isolated and cultured, and surface markers of hUC-MSCs were identified by flow cytometry. Forty Wistar male rats were divided into 4 groups: Control, Model, MSCs, Budesonide. The rats of Control group were sensitized and challenged by normal saline. The rats of Model, MSCs and Budesonide group were sensitized and challenged by ovalbumin (OVA) for 8 weeks. The MSCs group rats were given a tail vein injection of MSCs 0.2 ml (1×10(6) /ml) on days 35, 45, and 55 half an hour befor each OVA exposure. The Budesonide group rats were given aerosol inhalation of budesonide 2 mg two hours before each OVA exposure. Specimens were collected within 24 hours after the last OVA challenge. The parameters of airway morphological changes and the degree of airway remodeling were analyzed using Image-pro plus computer graphics. The levels of transformation growth factor (TGF) -ß(1) in bronchoalveolar lavage fluid (BALF) and serum were detected by enzyme linked immunosorbent assay (ELISA). The expressions of E-cadherin, α-smooth muscle actin (α-SMA) and fibronectin (Fn) were measured by immunohistochemistry. Results: The thickness of airway wall and smooth muscle of Model, MSCs and Budesonide group rats were significantly thicker than Control group. The levels of TGF-ß(1) in both BALF and serum of Model, MSCs and Budesonide group rats were significantly higher than Control group. The expression of E-cadherin of Model, MSCs and Budesonide group rats was significantly lower than Control group, while the expression of α-SMA and Fn were significantly higher. The thickness of airway wall and smooth muscle of MSCs and Budesonide group rats were significantly lower than Model group[(38.40±2.50, 45.34±0.33) vs (80.18±1.75) µm and (15.71±0.89, 18.57±0.67) vs (40.97±0.90) µm]. The levels of TGF-ß(1) in both BALF and serum of MSCs and Budesonide group were significantly lower than Model group[(3.53±0.43, 3.11±0.05) vs (20.88±0.37) µg/L and (31.07±0.89, 31.12±0.50) vs (70.58±0.39)µg/L](all P<0.01). The expressions of α-SMA, Fn of MSCs and Budesonide group rats were significantly lower than Model group[(0.438±0.057, 0.445±0.027) vs (0.521±0.030) and (0.459±0.041, 0.458±0.029) vs (0.527±0.022)], While the expression of E-cadherin was significantly higher[(0.308±0.023, 0.296±0.010) vs (0.256±0.087)](all P<0.01). Conclusion: MSCs could alleviate asthmatic airway remodeling, the mechanism of which may be associated with the inhibition of TGF-ß(1) induced epithelial-mesenchymal transition.

[Regulation effect of ß-catenin pathway on TGF-ß1 induced pulmonary pro-fibrosis].

OBJECTIVE: To investigate the regulation effect of ß-catenin pathway on transforming growth factor beta1 (TGF-ß1) induced pulmonary pro-fibrosis. METHODS: The rat alveolar typeⅡ cells (RLE-6TN) were divided into four groups: A1.control group; B1.TGF-ß1 group was treated with 5 µg/L TGF-ß1; C1.pcDNA+ TGF-ß1 group was transiently transfected with eukaryotic expression vector pcDNA3.0 (pcDNA) and followed by TGF-ß1 treatment (5 µg/L); D1.F-(ß-TrCP)-Ecad+ TGF-ß1 group was transiently transfected with ß-catenin protein knockout vector [F-(ß-TrCP)-Ecad] and followed by TGF-ß1 treatment (5 µg/L). After 24 hours, cells were observed under the inverted phase contrast microscope, then the expressions of E-cadherin, α-smooth muscle actin (α-SMA) and fibronectin (Fn) in each group were measured by Western blot and the mRNA levels of Snail which was the downstream profibrotic transcription production in cell culture supernatants of each group were detected by real-time fluorescence quantification-polymerase chain reaction (RT-PCR) .The rat alveolar macrophages (CRL-2192) were divided into five groups: A2.control group; B2.Interferon gamma (IFN-γ) group was treated by 20 µg/L IFN-γ; C2.TGF-ß1+ IFN-γ group was treated by 20 µg/L IFN-γ with 10 µg/L TGF-ß1; D2.F-(ß-TrCP)-Ecad+ TGF-ß1+ IFN-γ group was transfected with F-(ß-TrCP)-Ecad and other dispose was the same as group C2; E2.WTß-catenin+ TGF-ß1+ IFN-γ group was transfected with WTß-catenin and other dispose was the same as group C2.After 24 hours, protein levels of ß-catenin in group A2, B2, C2 were determined by Western blot.Inducible nitric oxide synthase (iNOS) mRNA levels of each group were detected by RT-PCR. RESULTS: The RLE-6TN cells of group B1, C1 showed a change in morphology to spindle-shaped cells, the cells of group D1 maintained a cobblestone morphology. Protein expressions of the fibroblast markers α-SMA and Fn, and mRNA expressions of the downstream profibrotic transcription production Snail of group B1, C1 were significantly higher than group A1, while protein expressions of the epithelial marker E-cadherin were significantly lower.The protein expressions of α-SMA, Fn and mRNA expressions Snail of group D1 were significantly lower than group C1 (0.352±0.076 vs 0.937±0.303, 0.319±0.072 vs 0.903±0.211, 3.675±0.642 vs 9.708±2.031), while the protein expressions of E-cadherin were significantly higher (1.482±0.227 vs 0.604±0.121) (all P<0.05). The steady state protein levels of ß-catenin in CRL-2192 cells was low and ß-catenin protein expressions of CRL-2192 cells in group A2, B2 and C2 had no significantly statistical differences.The mRNA expressions of iNOS of group B2 cells were significantly higher than group A2, C2, D2, E2 (64.95±4.47 vs 9.87±0.73, 21.32±2.41, 18.35±3.61, 22.87±3.14) (all P<0.01), the expressions of iNOS of group C2, D2, E2 were all higher than group A2 (all P<0.05), but there were no significant differences among group C2, D2 and E2. CONCLUSIONS: Inhibition of ß-catenin pathway inhibits TGF-ß1-induced epithelial-mesenchymal transition (EMT) and has no effect on its anti-inflammation effect.Therefore, ß-catenin pathway regulates the pulmonary pro-fibrosis effect of TGF-ß1.

The anatomical basis and prevention of neurogenic voiding dysfunction following radical hysterectomy.

The disorder of neurogenic dysfunction is one of the most important complications of radical hysterectomy. In order to prevent this potential complication, the authors have studied the composition and layers of the pelvic paravisceral structures. The nerve branching and distribution of the pelvic plexus of 12 adult female cadavers were analyzed. From lateral to medial the pelvic paravisceral structure is made up of three layers. The lateral layer is the pelvic visceral fascia, the middle, a vascular layer, and the medial one, a nervous one which consists of the pelvic plexus and subsidiary plexuses. The pelvic plexus and subsidiary plexuses are laid closely to the lateral walls of pelvic organs. The ischial spine was taken as the central point and two perpendicular lines penetrating through the ischial spine were used as the longitudinal axis and transverse axis. According to these landmarks, the pelvic plexus could be divided into three parts: behind the longitudinal axis are the roots of the pelvic plexus, near the longitudinal axis is the uterovaginal plexus, and in front of the longitudinal axis are the branches distributed to bladder and urethra. The pelvic plexus and the uterosacral and cardinal ligaments are closely related. The pelvic and subsidiary plexuses can be damaged in radical hysterectomy and voiding dysfunction may then develop. Some anatomic bases are provided to explain and hopefully prevent this from happening.