Palladium-Catalyzed Dearomative Heck/Suzuki Coupling of C2-Deuterated or C2-Nonsubstituted Indoles.

A palladium-catalyzed dearomative diarylation of C2-deuterated or C2-nonsubstituted indoles through domino Heck/Suzuki coupling is established. Relying on electron-deficient phosphite ligand, side reactions including intermolecular Suzuki coupling and intramolecular C-D/H arylation are inhibited and a wide range of 2,3-diarylated indolines bearing vicinal tertiary stereocenters including deuterated ones are afforded in moderate to excellent yields (up to 94%) and excellent diastereoselectivities (>20:1). The catalyst loading can be lowered to 0.02 mol % at elevated temperature.

Catalpol Enhances Random-Pattern Skin Flap Survival by Activating SIRT1-Mediated Enhancement of Autophagy.

Random-pattern skin flap necrosis limits its application in the clinic. It is still a challenge for plastic surgeons. Catalpol is an effective ingredient extracted from Rehmannia glutinosa, which is reported to promote angiogenesis and protect against ischemic cerebral disease. The aim of our experiment is to assess whether catalpol can facilitate random flap survival and the underlying mechanisms. Male "McFarlane flap" rat models were employed to explore the protective effects of catalpol. The range of necrosis in the flap was calculated 7 days after the models were established. The flap specimens were harvested for further experiments, including angiogenesis, apoptosis, oxidative stress, and autophagy evaluation. Catalpol-treated group promoted the average survival area of the flap than that in the control group. Based on immunohistochemical staining, Western blotting, and ROS detection, we found that catalpol significantly reduces oxidative stress and apoptosis and increases angiogenesis. Hematoxylin and eosin (H&E) staining and laser Doppler images further clarified the enhancement of angiogenesis after catalpol treatment. The impact of catalpol in flap was switched by using 3-methyladenine (3MA), proving the important role of autophagy in curative effect of catalpol on skin flaps. Importantly, the ability of catalpol to regulate autophagy is mediated by the activation of sirtuin 1 (SIRT1) based on its high affinity for SIRT1. Our findings revealed that catalpol improved the viability of random skin flaps by activating SIRT1-mediated autophagy pathway.

Apelin/APJ-Manipulated CaMKK/AMPK/GSK3ß Signaling Works as an Endogenous Counterinjury Mechanism in Promoting the Vitality of Random-Pattern Skin Flaps.

A random-pattern skin flap plays an important role in the field of wound repair; the mechanisms that influence the survival of random-pattern skin flaps have been extensively studied but little attention has been paid to endogenous counterinjury substances and mechanism. Previous reports reveal that the apelin-APJ axis is an endogenous counterinjury mechanism that has considerable function in protecting against infection, inflammation, oxidative stress, necrosis, and apoptosis in various organs. As an in vivo study, our study proved that the apelin/APJ axis protected the skin flap by alleviating vascular oxidative stress and the apelin/APJ axis works as an antioxidant stress factor dependent on CaMKK/AMPK/GSK3ß signaling. In addition, the apelin/APJ-manipulated CaMKK/AMPK/GSK3ß-dependent mechanism improves HUVECs' resistance to oxygen and glucose deprivation/reperfusion (OGD/R), reduces ROS production and accumulation, maintained the normal mitochondrial membrane potential, and suppresses oxidative stress in vitro. Besides, activation of the apelin/APJ axis promotes vascular migration and angiogenesis under relative hypoxia condition through CaMKK/AMPK/GSK3ß signaling. In a word, we provide new evidence that the apelin/APJ axis is an effective antioxidant and can significantly improve the vitality of random flaps, so it has potential be a promising clinical treatment.

Glycyrrhizin inhibits osteoarthritis development through suppressing the PI3K/AKT/NF-κB signaling pathway in vivo and in vitro.

Osteoarthritis (OA) is a serious and frequently occurring disease in the elderly, characterized by cartilage degeneration and proliferation of bone structure. Glycyrrhizin, a compound extracted from licorice, has been reported to have various important biological activities, such as antioxidant properties and anti-inflammatory action. However, it has not been reported whether glycyrrhizin has a positive effect on OA development. Our study aimed to evaluate the effects of glycyrrhizin on human OA chondrocytes. In the present study, we discovered that glycyrrhizin remarkably suppressed the interleukin (IL)-1ß-induced level of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) and the production of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOs), metalloproteinase3 (MMP3), metalloproteinase13 (MMP13) and a disintegrin and metalloproteinase with thrombospondin motifs5 (ADAMTS5). In addition, glycyrrhizin inverted the degradation of aggrecan and collagen II. Moreover, it significantly inhibited IL-1ß-stimulated PI3K/AKT phosphorylation and NF-κB mobilization in human OA chondrocytes. In vivo, glycyrrhizin treatment prevented the destruction of cartilage in mice OA models. In summary, all the results demonstrate that glycyrrhizin may be a potential therapeutic approach for OA.

Vasculature Characterization of a Multiterritory Perforator Flap: An Experimental Study.

Background Tip necrosis in the perforator flap is a significant problem in clinical practice. This study aimed to characterize the vasculature of a multiterritory perforator flap using a rat model and to investigate the impact of the vasculature on flap survival. Methods In total, 105 Sprague Dawley rats were divided into seven groups, including the control, 3 hours postoperative (PO), 12 hours PO, 1 day PO, 3 days PO, 5 days PO, and 7 days PO. A perforator flap with three territories based on the deep iliac circumflex artery was performed. Flaps with only skin incisions and vessel exposure were performed in the control group. The first choke zone (FCZ) was located between the anatomical and dynamic territories, and the second choke zone (SCZ) was located between the dynamic and potential territories. Sodium fluorescein and lead oxide-gelatin angiography and histological examination were performed in each group. Results Sodium fluorescein angiography revealed delayed staining in the perforator flap PO, particularly in the FCZ and SCZ. The delay phenomenon disappeared after 12 hours PO in the FCZ and after 1 day PO in the SCZ. Nonfluorescein-stained areas were found distal to the potential territory. In the FCZ PO, the choke vessels were dilated, while the number of microvessels was increased in the SCZ without choke vessel dilation. Conclusions The remodeling of choke vessels and increase in microvessel number represent arteriogenesis and angiogenesis, respectively. This neovascularization was responsible for flap survival in the entire dynamic territory and part of the potential territory.

Distribution of fibroblast growth factors and their roles in skin fibroblast cell migration.

Fibroblast growth factor (FGF)2/basic FGF is a member of the fibroblast growth factor family. Its function in skin wound healing has been well-characterized. However, the function of other FGFs in skin tissues remains to be elucidated. In the present study, FGF expression patterns in heart, liver, skin and kidney tissues were analyzed. Notably, in contrast to other tissues, only four FGFs, FGF2, 7, 10 and 21, were dominant in the skin. To examine FGF function in the wound healing process, mouse NIH3T3 fibroblast cells were treated with FGF2, FGF10 and FGF21, and cell migration was monitored. The results revealed that FGF treatment promoted cell migration, which is an important step in wound healing. In addition, FGF treatment enhanced the activity of c-Jun N-terminal kinase (JNK), a key regulator of fibroblast cell migration. To analyze its role in cell migration, FGF7 was overexpressed in fibroblast cells via a lentivirus system; however, this did not change cell migration speed. FGF2, 7, 10 and 21 were highly expressed in skin tissue, and all except FGF7 regulated fibroblast cell migration and activated JNK. The results of the present study increase our understanding of the role of FGFs in skin wound healing.

The Effect of Inducible Nitric Oxide Synthase on Multiterritory Perforator Flap Survival in Rats.

Background Inducible nitric oxide synthase (iNOS) plays an important role in vasodilation, angiogenesis, and ischemia-reperfusion injury. We investigated the effects of iNOS on the survival and choke vessels of multiterritory perforator flaps in rats. Methods In this study, 84 rats were divided into two groups of 42 rats each and subjected to multiterritory perforator flap operations. Rats in group A received daily intraperitoneal doses of 100 mg per kg of aminoguanidine (AG) and rats in group B received daily intraperitoneal injections of the same volume of saline solution. On postoperative day 7, the surviving flap area was calculated as a percentage of the total flap dimensions using DP2-BSW software. The diameter and density of microvessels in the second choke zone of the flap were calculated from histology studies. The nitric oxide (NO) content was measured using NO concentration assay kits, and the levels of vascular endothelial growth factor (VEGF) and iNOS were assessed using western blotting. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured using test kits. Laser Doppler imaging was used to evaluate flap perfusion in the second choke zone for 7 days after surgery. Results The flap survival area, diameter and density of microvessels, iNOS and VEGF levels, NO content, blood perfusion, and MDA content were significantly higher in the control group compared with the AG group, whereas SOD activity was significantly lower in the control group. Conclusions iNOS has a beneficial effect on the survival of multiterritory perforator flaps.

[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.

[Clinical research of vacuum sealing drainage combined with artificial dermis for the treatment of ankle and foot soft tissue defects].

OBJECTIVE: To investigate the effect of vacuum sealing drainage (VSD) combined with artificial dermis for treatment of the ankle and foot soft tissue defects. METHODS: A total of 15 patients with skin and soft tissue defect of ankle and foot were treated from January 2011 to December 2013, including 10 males and 5 females, with an average age of 32.5 years old ranging from 3 to 55 years old, involving 8 cases by traffic accident, 2 cases by machine accident, 5 cases by crush injury;8 cases with soft tissue exposure, 2 cases with tendor exposure, 5 cases with bone exposure. VSD was used to cover the wounds and continuous negative pressure drainage after debridement, the wounds covered with artificial dermis after the second granulation tissue grew well, again VSD was used to cover the wounds and negative pressure suction was applied, after 7 to 14 days negative pressure closed drainage was removed, free skin graft was transplanted above the artificial dermis, sterile gauze was used to compression bandage. RESULTS: All cases were followed up for 3 to 14 months with an average of 6.5 months. Skin graft of 15 of patients survived after transplantation, artificial dermis graft interval was 7 to 14 days with an average of 9.5 days. There was no obvious scar hyperplasia and contracture, no obvious pigmentation, appearance and functional recovery. CONCLUSIONS: After the implantation of artificial dermis and traditional skin graft method need for 2 to 3 weeks, vacuum sealing drainage technique combined with artificial dermis in treatment of soft tissue defect of foot and ankle skin has advantages of simple operation, significantly shorten the time of the second phase of the skin, without flap to repair, little injury to donor skin area, wound healing quality high, clinical effect of satisfaction.

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.

Correlation between E-cadherin-regulated cell adhesion and human osteosarcoma MG-63 cell anoikis.

PURPOSE: The aim of this study was to investigate the relationship between cell adhesion and anoikis evasion among human osteosarcoma cells (MG-63), and to further study the molecular mechanisms. MATERIALS AND METHODS: Human osteosarcoma cells (MG-63) were assessed for apoptosis, and caspase-3, E-cadherin and ß-catenin expression in EDTA and control non-EDTA groups. RESULTS: MG-63 cells were predominantly aggregated when in suspension, and the suspended cells were more dispersed in the EDTA group. Following culture in suspension for 24 h, 48 h, or 72 h, the rates of apoptosis were 34.88%±3.64%, 59.3%±7.22% and 78.5%±5.21% in the experimental group and 7.34%±2.13%, 14.7%±3.69%, and 21.4%±3.60% in the control group, respectively. Caspase-3 expression progressively increased and E-cadherin and ß-catenin were decreased in the experimental group, whereas there was no change in the control group. CONCLUSIONS: MG-63 cells could avoid anoikis through cell adhesion, and E-cadherin might play a role in this process.

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.

[Case-control study on therapeutic effects of different fixation methods for the treatment of olecranon fracture].

OBJECTIVE: To evaluate the clinical effects by comparing three different fixation methods: tension band, hollow lag screw and anatomical plate. METHODS: From January 2010 to January 2012, 82 patients with olecranon fractures who underwent surgical treatments were followed-up. All the patients were divided into three groups: tension band fixation group (group A), hollow lag screw fixation group (group B), anatomical plate fixation (group C). In group A, there were 35 patients, including 19 males and 16 females, ranging in age from 32 to 49 years old, with an average of (43.6 ± 8.7) years old, and the patients were treated with tension band fixation. According to Colton classification, there were 5 cases of type I, 3 cases of type II A,19 cases of type II B, and 8 cases of type II C in group A. Among 20 patients in group B, there were 13 males and 7 females, ranging in age from 27 to 50 years old, with an average of (41.5 ± 9.3) years old. The patients in group B were treated with hollow lag screw fixation. According to Colton classification, there were 4 cases of type I, 4 cases of type II A, and 12 cases of type II B in group B. In group C, there were 27 patients totally, including 15 males and 12 females, ranging in age from 30 to 55 years old, with an average of (38.2 ± 6.2) years old. The patients in group C were treated with anatomical plate fixation. According to Colton classification, there were 4 cases of type II B, 13 cases of type II C, and 10 cases of type II D in group C. The Fracture healing time, complications and functional recovery were retrospectively observed and recorded. RESULTS: All the patients were followed up, and the duration ranged from 8 to 24 months, with an average of 15 months. The average healing time of patients in group C was the longest among three groups. The flexion-extension and rotation activities of elbow joint in group B and C were better than that in group C. According to Broberg & Morrey score system, the therapeutic effects of patients in group A and B were better than that of group C. In group C, 2 patients had incision infections, 6 patients complained of foreign body sensation, 1 patient got a delayed fracture healing, and 1 patient had the heterotopic ossification. There were no occurrences of incision infections in group A and B; internal fixation loosening occurred in 3 patients in group A and 2 patients in group B; delayed fracture healing occurred in 2 patients in group A and 2 patients in group B; and skin bursa formation occurred in 6 patients in group A and 1 patient in group B. CONCLUSION: All the three ways are effective methods for the treatment of olecranon fractures. Fixation methods should be selected depending on the type of fracture.

[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.

[Effects of agonists of PPAR-gamma on angiotensin II-induced proliferation and extracellular matrix synthesis in hypertrophic scar fibroblasts].

OBJECTIVE: To study the effects of peroxisome proliferator-activated receptor gamma agonists on angiotensin II-induced cellular response in cultured fibroblasts derived from patients with hypertrophic scars, so as to investigate its effects on preventing the formation of hypertrophic scars. METHODS: Fibroblasts were freshly isolated from hypertrophic scars and cultured with angiotensin II, rosiglitazone and GW9662 at a certain concentration. Fibroblasts proliferation were assessed via Cell Counting Kit-8; the mRNA and protein expressions of Collagen I and Fibronectin (FN) were determined by quantitative real-time RT-PCR and Western blotting. RESULTS: The absorbance of CCK-8 and relative expression of Collagen I, FN mRNA and protein were 1.082 5 +/- 0.007, 6.45 +/- 0.97, 4.92 +/- 0.86, 2.92 +/- 0.41, 2.78 +/- 1.04 in Ang II group; 0.722 4 +/- 0.012, 1.82 +/- 0.34, 1.78 +/- 0.27, 1.57 +/- 0.46, 1.68 +/- 0.39 in Ros + Ang II group; 0.554 7 +/- 0.012, 0.97 +/- 0.12, 1.07 +/- 1.08, 1.05 +/- 0.43, 1.14 +/- 0.36 in Ros group; 1.056 0 +/- 0.005, 5.83 +/- 0.24, 4.47 +/- 0.32, 2.69 +/- 0.35, 2.62 +/- 0.27 in GW9662 + ros + Ang II group. The results showed a significant difference between the Ang II group and the control group (P < 0.05). The effect of Ang II could be markedly inhibited by Ros (P < 0.05). In addition, Ros did not influence cell proliferation and production of extracellular matrix (P > 0.05). There was a significant difference between the GW9662 + Ros + Ang II group and the Ros + Ang II (P < 0.05). CONCLUSIONS: PPAR-gamma agonists inhibit Ang II-induced proliferation and extracellular matrix synthesis effectively in the hypertrophic scar fibroblasts. Thus PPAR-gamma agonists may have potential therapeutic effect for hypertrophic scar.

Functional motor nerve regeneration without motor-sensory specificity following end-to-side neurorrhaphy: an experimental study.

PURPOSE: To evaluate the quality of regenerating myelinated axons and motor-sensory specificity in an end-to-side nerve repair model. METHODS: We divided 20 rats into 3 groups: (1) end-to-side neurorrhaphy using the ulnar nerve as donor nerve and the musculocutaneous nerve as recipient nerve; (2) normal control; and (3) transected nerve with the stumps buried. At 5 months, we monitored the grooming test, the electrophysiological response, and the histologic changes in nerve and muscle. RESULTS: Grooming recovered successfully, and electrophysiological investigations revealed that the target muscles had been reinnervated in the end-to-side group. The mean wet weight of the reinnervated biceps brachii muscle was 72% of the normal muscle, and the mean muscle fiber cross-sectional area of the reinnervated muscle was similar to the normal muscle. The implanted musculocutaneous nerve contained varying but satisfactory numbers of axons (end-to-side group: 596 ± 348 vs normal group: 1,340 ± 241). Acetylcholinesterase staining revealed a similar percentage of myelinated fibers in the musculocutaneous nerve (39%) and the biceps brachii branch of the musculocutaneous nerve (38%) in the end-to-side group. This was similar to the number of myelinated fibers in the donor ulnar nerve (37%). CONCLUSIONS: The present study confirms that limited but functional reinnervation can occur on the basis of collateral sprouting of intact axons from the ulnar nerve. The motor-sensory specificity is not important.

[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.