[Acupuncture reduce colonic inflammation by suppressing oxidative stress and endoplasmic reticulum stress in rats with ulcerative colitis].

OBJECTIVE: To investigate the effect of manual acupuncture (MA) and electroacupuncture (EA) on histopathological changes, and levels of oxidative-stress related cytokines and key proteins of the endoplasmic reticulum stress (ERS) pathway in ulcerative colitis (UC) rats, so as to reveal their mechanisms underlying improvement of UC. METHODS: Twenty-eight male SD rats were randomly divided into control, model, EA and MA groups (n＝ 7 rats per group). The UC model was established by enema of mixture solution of 5% 2, 4, 6-trinitrobenzene sulfonic acid (TNBS, 100 mg/kg). Rats of the control group received intra-rectal perfusion of normal saline. After modeling, the left "Quchi"(LI11) and "Zusanli"(ST36) were stimulated with EA (2-4 mA，8 Hz/25 Hz) or MA for 20 min, once every other day for consecutive 2 weeks. The rats in the control and model group were just anesthetized and fixed. At the end of experiments, the colon tissue was collected for observing histopathological changes with H．E. staining. The contents of oxidative stress-related factors as superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), reduced glutathione (GSH) and total antioxidant capacity (T-AOC) were detected by ELISA, and the expression levels of key proteins of ERS as phosphorylated inhibitor of nuclear factor kappa B kinase α (p-IκBα), phosphorylated p65 (p-p65), glucose regulated protein 78 (GRP78), phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK) and phosphorylated eukaryotic translation initiation factor 2α (p-eIF2α) by using Western blot. RESULTS: After modeling, the colon tissues showed severe swelling, disordered arrangement of intestinal mucosal cells, hemorrhage with infiltration of inflammatory cell and partial loss of colon villus, which was relatively milder in the EA and MA groups. The colonic lesion score was remarkably increased in the model group in contrast to the control group (P<0.01), and obviously reduced in both EA and MA groups relevant to the model group (P<0.01). The levels of SOD, CAT, GSH and T-AOC were all significantly decreased (P<0.01), and the content of MDA, and expression levels of p-IκBα， p-p65 and GRP78, p-PERK and p-eIF2α proteins were all significantly increased in the model group relevant to the control group (P<0.01). After the treatment, modeling-induced down-regulation of SOD, CAT and GSH in both EA and MA groups, and T-AOC in the EA group, and up-regulation of levels of MDA, p-IκBα， p-p65, GRP78, p-PERK and p-eIF2α in both groups were reversed (P<0.01, P<0.05). CONCLUSION: Both EA and MA treatment can obviously alleviate colonic inflammation in UC rats via inhibiting oxidative stress and ERS.

Role of Homeodomain-Interacting Protein Kinase 2 in the Pathogenesis of Tissue Fibrosis in Keloid-Derived Keratinocytes.

Epithelial-mesenchymal transition (EMT) plays a critical role in fibrotic keloid formation, which is characterized by excessive collagen and extracellular matrix synthesis and deposition. Growing evidence suggests that the serine/threonine kinase homeodomain-interacting protein kinase 2 (HIPK2) acts upstream of several major fibrosis signaling pathways; however, the role of HIPK2 in the keloid fibrogenesis remains unknown. In the current study, we investigated the roles of HIPK2 in the pathogenesis of keloids. Primary normal skin and keloid keratinocytes were cultured and pretreated with transforming growth factor (TGF)-ß1. Next, keratinocytes were transfected with scrambled small interfering RNA (siRNA) and anti-HIPK2 siRNA. The TGF-ß1-associated HIPK2 alterations were investigated by quantitative real-time polymerase chain reaction. Protein levels were analyzed by western blotting. The HIPK2 was markedly increased in the keloid-derived keratinocytes compared with normal skin keratinocytes. In addition, HIPK2 induced the expression of EMT markers in normal skin keratinocytes by TGF-ß1-SMAD family member 3 (SMAD3). The effect of TGF-ß1-related EMT markers and SMAD3 phosphorylation in response to added TGF-ß1 was significantly abrogated when the cells were transfected with HIPK2 siRNA. We conclude that HIPK2 is a crucial factor in the pathogenesis of keloids, suggesting that HIPK2 might be a novel potential drug target for antikeloid therapy.

Nail Fusion Plasty: Nail Cosmetic Results and Assessment Criteria of Nail Reconstruction.

PURPOSE: This study aimed to develop a scoring system for evaluating the nail appearance after nail fusion plasty of the duplicated thumb, and to investigate the operation indication of the combination procedure and its effect on the postoperative nail appearance. METHODS: The nail fusion plasty technique was carried out in 20 patients with congenital thumb duplication. The patients were followed up to observe the appearance of nail, and the results were assessed by our nail appearance evaluation criteria. RESULTS: Twenty patients were followed up for 18 to 92 months (48 months on average). According to our criteria, we achieved excellent results in 12 cases, good in 6 cases, fair in 1 case, and poor in 1 case. CONCLUSIONS: The combination procedure is recommended when the size of the thumb is less than 80% of the size of the normal side or the width of the nail is less than 80% of that of the normal side. Satisfactory results can be obtained by careful operation. The newly devised scoring system assesses the nail appearance in a very comprehensive manner.

NADPH oxidase-2 is a key regulator of human dermal fibroblasts: a potential therapeutic strategy for the treatment of skin fibrosis.

The proliferation of human skin dermal fibroblasts (HDFs) is a critical step in skin fibrosis, and transforming growth factor-beta1 (TGF-ß1) exerts pro-oxidant and fibrogenic effects on HDFs. In addition, the oxidative stress system has been implicated in the pathogenesis of skin disease. However, the role of NADPH oxidase as a mediator of TGF-ß1-induced effects in HDFs remains unknown. Thus, our aim was to investigate the role of NADPH in human skin dermal fibroblasts. Primary fibroblasts were cultured and pretreated with various stimulants. Real-time Q-PCR and Western blotting analyses were used for mRNA and protein detection. In addition, siRNA technology was applied for gene knock-down analysis. Hydrogen peroxide production and 2',7'-dichlorofluorescein diacetate (DCFDA) measurement assay were performed. Here, our findings demonstrated that HDFs express key components of non-phagocytic NADPH oxidase mRNA. TGF-ß1 induced NOX2 and reactive oxygen species formation via NADPH oxidase activity. In contrast, NOX3 was barely detectable, and other NOXs did not display significant changes. In addition, TGF-ß1 phosphorylated MAPKs and increased activator protein-1 (AP-1) in a redox-sensitive manner, and NOX2 suppression inhibited baseline and TGF-ß1-mediated stimulation of Smad2 phosphorylation. Moreover, TGF-ß1 stimulated cell proliferation, migration, collagen I and fibronectin expression, and bFGF and PAI-1 secretion: these effects were attenuated by diphenylene iodonium (DPI), an NADPH oxidase inhibitor, and NOX2 siRNA. Importantly, NOX2 siRNA suppresses collagen production in primary keloid dermal fibroblasts. These findings provide the proof of concept for NADPH oxidase as a potential target for the treatment of skin fibrosis.

[Effect of hirudin on random skin flap survival in rats].

OBJECTIVE: To investigate the effect of Hirudin on random skin flap survival in rats. METHODS: 24 SD rats were randomly divided into control group and experimental group. The "McFarlane flap (3 cm x 9 cm)" rat models were established on the rat dorsum. 3 ml Hirudin (30 ATU) was injected into the flap in the experimental group, while 3 ml saline in the control group. The injection was performed for 7 days. The flap survival area in the two groups was measured. The tissue samples were taken from proximal (I), middle (II) and distal (III) portions of flaps for histologic study. The VEGF and bFGF expression was also detected with immunohistochemistry method. RESULTS: 7 days after operation, the flap survival rate was (69.52 +/- 3.23)% in the experimental group, while (50.36 +/- 2.37)% in control group, showing a significant difference between the two groups (P < 0. 01). In the middle portion, tissue edema and infiltration of neutrophils in experimental group was markedly slighter than that in control group. The VEGF and bFGF expression and neovascularization was enhanced markedly in experimental group. CONCLUSIONS: Hirudin can increase the survival of random pattern skin flaps. It may increase the VEGF, bFGF expression through a series of complex regulatory pathway. Then flap neovascularization is promoted and the flap blood supply is increased.

[Application of dorsal pentagonal flap for reconstruction of the web space in congenital syndactyly].

OBJECTIVE: To investigate the therapeutic effect of dorsal pentagonal flap for reconstruction of the web space in congenital syndactyly. METHODS: From October 2007 to August 2009, 10 patients with congenital syndactyly were treated with dorsal pentagonal flap for web space reconstruction. During the follow-up period, the patients underwent finger functional assessment and web space appearance examination. The satisfactory rate of patients was also investigated. RESULTS: 10 patients with 17 reconstructed web spaces were followed up for 4-25 months (median, 15 months). Normal passive flexion and extension of fingers were achieved with good cosmetic result at web space. The abduction angle of fingers ranged from 30 degrees to 45 degrees. No web creep occurred and no secondary procedures were required. All the parents of the patients were satisfied with the results. CONCLUSIONS: Web space reconstruction with dorsal pentagonal flap is easily performed with reliable blood supply and low re-operated rate. The cosmetic and functional result is satisfactory.