Overexpression of EphB2 in the basolateral amygdala is crucial for inducing visceral pain sensitization in rats subjected to water avoidance stress.

AIMS: Basolateral amygdala (BLA), as a center for stress responses and emotional regulation, is involved in visceral hypersensitivity of irritable bowel syndrome (IBS) induced by stress. In the present study, we aimed to investigate the role of EphB2 receptor (EphB2) in BLA and explore the underlying mechanisms in this process. METHODS: Visceral hypersensitivity was induced by water avoidance stress (WAS). Elevated plus maze test, forced swimming test, and sucrose preference test were applied to assess anxiety- and depression-like behaviors. Ibotenic acid or lentivirus was used to inactivate BLA in either the induction or maintenance stage of visceral hypersensitivity. The expression of protein was determined by quantitative PCR, immunofluorescence, and western blot. RESULTS: EphB2 expression was increased in BLA in WAS rats. Inactivation of BLA or downregulation of EphB2 in BLA failed to induce visceral hypersensitivity as well as anxiety-like behaviors. However, during the maintenance stage of visceral pain, visceral hypersensitivity was only partially relieved but anxiety-like behaviors were abolished by inactivation of BLA or downregulation of EphB2 in BLA. Chronic WAS increased the expression of EphB2, N-methyl-D-aspartate receptors (NMDARs), and postsynaptic density protein (PSD95) in BLA. Downregulation of EphB2 in BLA reduced NMDARs and PSD95 expression in WAS rats. However, activation of NMDARs after the knockdown of EphB2 expression still triggered visceral hypersensitivity and anxiety-like behaviors. CONCLUSIONS: Taken together, the results suggest that EphB2 in BLA plays an essential role in inducing visceral hypersensitivity. In the maintenance stage, the involvement of EphB2 is crucial but not sufficient. The increase in EphB2 induced by WAS may enhance synaptic plasticity in BLA through upregulating NMDARs, which results in IBS-like symptoms. These findings may give insight into the treatment of IBS and related psychological distress.

Modified autologous adipose transplantation in the treatment of depressed scars: an experimental study.

OBJECTIVE: Complex depressed scars can cause tissue adhesion, resulting in serious joint dysfunction. In recent years, autologous adipose and adipose-derived stem cells have been widely used to treat depressed scars, but there are still limitations in these treatment that should be resolved. This study aimed to investigate the therapeutic effects of adipose tissues collected with modified technique on the depressed scars in animals. METHODS: The adipose tissues were collected with a forward technique, and tissue viability in vitro and the survival of transplanted tissues in in nude mice were further assessed. Furthermore, the therapeutic effects of adipose tissues collected with new technique and traditional technique on the depressed scars were explored in an animal model of bleomycin induced scar formation. RESULTS: The adipose tissues collected with the new technique had a higher glucose transport (P<0.01); after transplantation into the nude mice, the amount of residual tissues and the survival rate in the modified group were higher than in the traditional group (P<0.05); electron microscopy showed the intercellular space was covered with reticular structure, in which there was a large amount of microvessel structure in the adipose tissue of the modified group; immunohistochemistry showed that the microvessel density (MVD) in the modified group increased significantly (P<0.01). At 28 d after transplantation into the scar animals, the dermal collagen fibers became thicker and showed regular arrangement, the myofibroblasts became regenerative and inflammation was improved as compared to blank control group. In the untreated scar group, the collagen fibers were loose and irregular, and a large amount of inflammatory cells was observed. In addition, the dermal expression of α-SMA and TGF-ß1 in the transplantation group reduced significantly as compared to scar group (P<0.05). CONCLUSION: The autologous adipose tissues collected with the new technique possess higher activity ad contain more. In scar animals, transplantation of these adipose tissues may improve the scar structure and inhibit the scar formation which may be related to the suppressed expression of α-SMA and TGF-ß1.

Altered distributions of bone tissue mineral and collagen properties in women with fragility fractures.

Heterogeneity of bone tissue properties is emerging as a potential indicator of altered bone quality in pathologic tissue. The objective of this study was to compare the distributions of tissue properties in women with and without histories of fragility fractures using Fourier transform infrared (FTIR) imaging. We extended a prior study that examined the relationship of the mean FTIR properties to fracture risk by analyzing in detail the widths and the tails of the distributions of FTIR properties in biopsies from fracture and non-fracture cohorts. The mineral and matrix properties of cortical and trabecular iliac crest tissue were compared in biopsies from women with a history of fragility fracture (+Fx; n=21, age: mean 54±SD 15y) and with no history of fragility fracture (-Fx; n=12, age: 57±5y). A subset of the patients included in the -Fx group were taking estrogen-plus-progestin hormone replacement therapy (HRT) (-Fx+HRT n=8, age: 58±5y) and were analyzed separately from patients with no history of HRT (-Fx-HRT n=4, age: 56±7y). When the FTIR parameter mean values were examined by treatment group, the trabecular tissue of -Fx-HRT patients had a lower mineral:matrix ratio (M:M) and collagen maturity (XLR) than that of -Fx+HRT patients (-22% M:M, -18% XLR) and +Fx patients (-17% M:M, -18% XLR). Across multiple FTIR parameters, tissue from the -Fx-HRT group had smaller low-tail (5th percentile) values than that from the -Fx+HRT or +Fx groups. In trabecular collagen maturity and crystallinity (XST), the -Fx-HRT group had smaller low-tail values than those in the -Fx+HRT group (-16% XLR, -5% XST) and the +Fx group (-17% XLR, -7% XST). The relatively low values of trabecular mineral:matrix ratio and collagen maturity and smaller low-tail values of collagen maturity and crystallinity observed in the -Fx-HRT group are characteristic of younger tissue. Taken together, our data suggest that the presence of newly formed tissue that includes small/imperfect crystals and immature crosslinks, as well as moderately mature tissue, is an important characteristic of healthy, fracture-resistant bone. Finally, the larger mean and low-tail values of mineral:matrix ratio and collagen maturity noted in our -Fx+HRT vs. -Fx-HRT biopsies are consistent with greater tissue age and greater BMD arising from decreased osteoclastic resorption in HRT-treated patients.

Treatment for displacement of PAAG mixture after injection augmentation mammoplasty.

BACKGROUND: The incidence of serious complications after augmentation mammaplasty with injection of polyacrylamide hydrogel (PAAG) was high. OBJECTIVE: To design a new method for healing of the cavities and cysts after augmentation mammaplasty. METHODS: 102 patients in whom PAAG exceeded the breast and spread to the thoracic-abdominal walls were enrolled and divided into two groups. RESULTS: The flowing masses of different sizes exceeded the breast and spread to the thoracic-abdominal walls, and a large number of PAAG showed flowing degenerative mixture in the tissues and were invaded by many inflammatory cells. PAAG deposited extensively in the breast tissues, armpits and space of the thoracic-abdominal wall, and the breast was connected with the abdominal wall through the fistula of different sizes. At 2 weeks, the percentages of decrease in drainage volume and in lesion lacuna size of the thoracic-abdominal wall (82% and 80%, respectively) in patients receiving the multiple incisions combined with radical therapy were significantly different from those who did not receive the multiple incisions (46% and 45%) (Both P<0.01). At 4 weeks, in some of the patients receiving the multiple incisions combined with radical therapy, the lacuna of the thoracic-abdominal wall disappeared completely, and the lesions with flowing masses had been cleared. CONCLUSIONS: The new method of subareolar incision combined with surgery for inferior segment of mass to clean the mixture and thoroughly eliminate the lacuna of the thoracic-abdominal wall as well as suture to close the intramammary fistula can improve the treatment efficacy.

Multiscale contribution of bone tissue material property heterogeneity to trabecular bone mechanical behavior.

Heterogeneity of material properties is an important potential contributor to bone fracture resistance because of its putative contribution to toughness, but establishing the contribution of heterogeneity to fracture risk is still in an incipient stage. Experimental studies have demonstrated changes in distributions of compositional and nanomechanical properties with fragility fracture history, disease, and pharmacologic treatment. Computational studies have demonstrated that models with heterogeneous material properties predict apparent stiffness moderately better than homogeneous models and show greater energy dissipation. Collectively, these results suggest that microscale material heterogeneity affects not only microscale mechanics but also structural performance at larger length scales.

[Preparation of Rubber Accelerator Tetramethyl Thiuram Monosulfide and Its Spectral Analysis].

In the study, rubber accelerator tetramethyl thiuram monosulfide (TMTM) was synthesized firstly. Single crystal TMTM was cultivated by solvent evaporation method. TMTM was detected and characterized by XRD single crystal diffraction, FTIR, TG-DSC. The micro-structure and intrinsic regularity were revealed. The TMTM microstructure was revealed by XRD single crystal diffraction from the diffraction data, part of bond length and bond angle. Its highly efficient performance of rubber vulcanization promotion was decided due to its orientation structure and high order. Properties of TMTM was disclosed by TG-DSC analysis from the heat effect. The results of TG-DSC and FTIR showed that single crystal TMTM possessed with little CS2. The chemical bond types in TMTM were revealed by FTIR the same as single crystal diffraction testing by different way. The decomposition temperature of TMTM was very high. It could provided reference with research on rubber vulcanizing properties by TMTM on rubber vulcanizing machine. This study can help the enterprises to designate the working standard tracing detection of TMTM industrialized production. Performance index of TMTM was judged. The project of TMTM industry standard can be declared by the enterprises, written a draft standard. It can provide the basis experimental data on completing the revision of the national ministry of industry and information technology standard system project plan.

Comparison of Artecoll, Restylane and silicone for augmentation rhinoplasty in 378 Chinese patients.

PURPOSE: Dermal fillers have been proven to be safe in soft tissue augmentation; however, their efficacy in modeling the noses of Asian patients has not been demonstrated. METHODS: In this study, 378 patients were included and underwent augmentation rhinoplasty (Artecoll, n=126; Restylane, n=126 and silicone implants, n=126). The subjective and objective outcomes were evaluated on day 1, and months 1, 3, 6, and 12 after injection rhinoplasty. RESULTS: All patients achieved significant improvement in nasal shape and contour immediately after surgery. Patients treated with Restylane failed to maintain the nasal shape and contour 1 year after surgery, whereas patients undergoing Artecoll rhinoplasty completely maintained the post-treatment nasal shape and contour. More patients with silicone implants experienced adverse events and the severity of these events was greater in the silicone group compared to those in the Restylane and Acetoll groups. CONCLUSION: Artecoll rhinoplasty has a low incidence of adverse effects and the shape and contour of the nose are maintained for a prolonged period.

Smac/DIABLO regulates the apoptosis of hypertrophic scar fibroblasts.

In abnormal skin wound healing, hypertrophic scars (HS) are characterized by excessive fibroblast hypercellularity and an overproduction of collagen, leading to atypical extracellular matrix (ECM) remodeling. Although the exact mechanisms of HS remain unclear, decreased HS fibroblast (HSFB) apoptosis and increased proliferation are evident in the development of HS. In this study, the contribution of the second mitochondria-derived activator of caspases/direct inhibitor of apoptosis protein (IAP)-binding protein with a low isoelectric point (pI) (Smac/DIABLO), an apoptosis-promoting protein released from the mitochondria, was investigated in human normal skin and HSFB cultures. The expression of Smac/DIABLO is usually decreased in many malignant tumors compared with normal tissues. Immunohistochemical analysis of skin tissues and the western blot analyses of fibroblasts revealed that the expression of Smac/DIABLO was lower in HS tissues compared with normal skin tissues. Of note, adenovirus-mediated Smac/DIABLO overexpression in the cultured HSFBs significantly reduced cell proliferation, as detected by the cell counting kit-8, and increased caspase-3 and -9 activity, as detected by spectrofluorimetry. In addition, it increased apoptosis, as detected by fluorescence-activated cell sorting (FACS). Furthermore, we found that the silencing of Smac with siRNA in the HSFBs induced a noticeable decrease in caspase-3 and -9 activity, leading to a significant reduction in apoptosis. In addition, the mRNA expression of type I and III pro-collagen detected in the HSFBs was significantly increased following the silencing of Smac with siRNA and was inhibited following Smac/DIABLO overexpression, as shown by real-time RT-PCR. In conclusion, Smac/DIABLO decreases the proliferation and increases the apoptosis of HSFBs. To our knowledge, the data from our study suggest for the first time that Smac/DIABLO is a novel therapeutic target for HS.

Polyacrylamide hydrogel injection for augmentation mammaplasty: loss of ability for breastfeeding.

Polyacrylamide hydrogel (PAAG) has been widely used for injection augmentation mammaplasty in Russia, China, and Iran for more than 2 decades. In recent years, it has been advocated as a safe permanent filler for soft-tissue augmentation. However, the complications associated with PAAG injection in soft-tissue augmentation have not been extensively investigated. Augmentation mammaplasty through PAAG injection is associated with some complications. The incidence of infection during breastfeeding was reported to be higher than 50%. Herein, we report 58 cases of infection in breastfeeding women receiving PAAG injection, including 50 with unilateral injection (36 on the right, 14 on the left) and 8 bilateral injection. They experienced large breast autoinflation and some severe symptoms, such as local and systemic fever, breast swelling, nipple bulging, tenderness, and pain, which lead to surgical removal of galactocele or intraprosthetic collection of sterile pus resulting in deformity. Operation and comprehensive measures including removal of the injected material, clearing residual cavity, and pharmacotherapy were carried out to control infection and inflammation for 1 to 2 weeks. In the following 12 months, no relapse or recurrence of residual cavity was noted. Therefore, we do not recommend PAAG injection for augmentation mammaplasty, especially in women intending to breastfeed. Patients undergoing PAAG injection for augmentation mammaplasty should avoid breastfeeding. PAAG injection will cause serious consequences resulting in tissue atrophy and breast resection if inappropriately handled.

Autologous nerve implantation into denervated monkey skin promotes regeneration of Meissner's corpuscle.

BACKGROUND: The aim of this study was to observe the effects of autologous nerve implantation into the denervated finger flap on the regression and regeneration of sensory nerve endings and Meissner's corpuscles. MATERIAL/METHODS: Bilateral nerves of fingers were separated: one was removed and the other was implanted into the denervated finger in the implantation group. In the non-implantation group, both nerves were removed. The ventral skin of fingers was collected for immunohistochemistry and electron microscopy 3, 6, 9 and 12 months after surgery. RESULTS: The nerve endings in the Meissner's corpuscles began to degenerate 3 months after denervation. The elementary structure of Meissner's corpuscles was not significantly altered. Nerve fibers were present around the Meissner's corpuscles, accompanied by growing into its inward. The axons in the denervated nerve disappeared and the Meissner's corpuscles began to atrophy at month 6. More regenerated nerve fibers were observed after nerve implantation, including intensive and thick fibers, accompanied by reinnervation of Meissner's corpuscles. More nerve fibers and a higher proportion of myelinated nerve fibers were noted at month 9 in the implantation group, and the reinnervation was present in the majority of Meissner's corpuscles. Naive myelinated nerve fibers appeared at the caudal end of Meissner's corpuscles. The nerve fibers in the Meissner's corpuscles increased to the normal level at 12 months after nerve implantation. CONCLUSIONS: The implanted nerve regenerated a large amount of free nerve endings, which helped to regenerate simple Meissner's corpuscles via governing previously degenerated corpuscles.

[Significance of the secreted frizzled-related protein 2 expression in earlobe keloid].

OBJECTIVE: To investigate the expression and the role of secreted frizzled-related protein 2 (SFRP2) in the earlobe keloid and find a valid way to treat the keloid with gene therapy. METHODS: The expression of SFRP2 mRNA and protein was tested with in situ hybridization and Western Blot Analysis method in the different period of earlobe keloid. RESULTS: The SFRP2 mRNA and protein expression at the keloid edge was significantly high in 12 month group than in 3 or 6 month groups (P < 0.01), but not than in 24 month group. The SFRP2 expression started to decrease in the keloid center 12 month later (P < 0.01). The SFRP2 expression was always higher in edge than in center during all the period (P < 0.05, P < 0.01). CONCLUSIONS: The results suggest that SFRP2 may play an important role in the development of keloid, especially at the keloid edge. The high SFRP2 expression in endothelial cells and surrounding tissue is also important. It may be a new way for gene therapy of keloid by decreasing the SFRP2 expression.

[The interaction between secreted frizzled-related protein 2 and osteoblast-specific factor 2 in keloid].

OBJECTIVE: To verify the interaction between secreted frizzled-related protein 2 (SFRP2) and osteoblast-specific factor 2 (OSF-2). METHODS: HA-tagged OSF-2 fusion protein recombinant vector pCMV-HA-OSF-2, which could express in mammal cells was constructed, then identified by enzyme-cutting and transfected into human kidney 293 (HK293) cells with or without Myc-SFRP2 recombinant eukaryotic expression vector pCMV-HA-SFRP2. The interaction between SFRP2 and OSF-2 was detected through coimmunoprecipitation and Western blotting. RESULTS: In electrophoresis bath, target fragment of SFRP2 coding gene with 800 bp and target gene OSF-2 with 2500 bp could be seen respectively after enzyme-cutting, which showed that pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 were constructed successfully. No HA-OSF-2 expression was detected after pCMV-Myc-SFRP2 or pCMV-HA-OSF-2 transfection. Whereas, HA-OSF-2 expressed by Myc antibody immunoprecipitation after pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 co-transfection. CONCLUSIONS: HA-OSF-2 recombinant vector can express in mammal cells. Interaction exists between HA-OSF-2 and SFRP2.

[Influence of substance P on the release of histamine in the human hypertrophic scar tissue].

OBJECTIVE: To investigate the influence of substance P (SP) on the release of histamine in the human hypertrophic scar tissue, and to explore the prerequisite of their interaction. METHODS: Tissue specimens of normal skin and hypertrophic scar from eight hospitalized patients were excised and cut into 0.5 to 1.0 mm3 pieces, and the histamine release by mast cell (MC) under the stimulation of different concentration of SP and calcium, as well as the different affect time of SP, were determined with fluorescence spectrometer. Then the histamine release rate was calculated. RESULTS: There was obvious release of histamine when SP concentration was 1 x 10(-6) mol/L , and the release rate was (50.0 +/- 3.6) %, which was significantly higher than that by SP in the concentration of 0 mol/L [(44.0 +/- 3.2) %, P < 0.01]. Therefore it seemed to be dose-dependent. About 90% of histamine was released within 15 minutes of 5 x 10(-1) mol/L Substance P stimulation, and it was also time-dependent. The histamine release reached the peak when calcium concentration was 5 x 10(-3) mol/L, which seemed to be dose-dependent, but it decreased transiently when calcium concentration was 1 x 10(-3) mol/L. In all occasions, the influence of SP on the histamine release by MC in hypertrophic scar (HS) was markedly higher than that in normal skin (NS) (P < 0.01). Conclusion The influence of SP on the histamine release by MC in HS was markedly higher than that in NS, and it might be closely related to itching sensation and the formation of hypertrophic scar.

[Establishment of an animal model of human hyperplastic scar in nude mice].

OBJECTIVE: To establish an ideal model of human hyperplastic scar (HS) in nude mice, so as to provide us a new model to carry out further studies on the mechanism of HS development. METHODS: Full skin defect sized 2.0 cm x 1.5 cm was created on the back of 100 nude mice. The defect was thereafter covered with full thickness human skin. After the grafted skin survived, the nude mice were subjected to deep partial thickness burn of the grafted skin with heated copper rod. The development of the hyperplasia of the scar after wound healing was observed histologically and grossly. RESULTS: Grafted full-thickness human skin took and survived well in 86 out of 100 nude mice. There was obvious and continuous hyperplasia of scar in 67 mice (78%). The external appearance and histological features of the HS appeared similar to those in human HS. The average thickness of the scar was 0.34 cm, with the thickest part measuring 0.6 cm. In addition, the time of hyperplastic change lasted for 63 - 217 days in average of 128 days. CONCLUSION: Obvious and continuous scar hyperplasia could be found in this model, and the whole process beginning from wound healing to the formation of HS could be easily observed. The model was therefore suitable and ideal for the study of HS.

[Effect of combined application of external cyclosporine A and CTLA4Ig on the survival of rat auricle allograft].

OBJECTIVE: To study the effect of combined application of external cyclosporine A (CsA) and CTLA4Ig in inhibiting rejection and inducing immune tolerance in composite tissue allograft. METHODS: The auricles with vessel pedicle were transplanted from Lewis rats to BN rats under microsurgery. CsA was spreaded on the surface of grafts in combination with administration of CTLA4Ig intraperitoneally after transplantation. The rejection response and survival time of grafts were observed, and the IL-2 level in serum was measured. RESULTS: The mean survival time was (7.8 +/- 1.7) days in control group. It was (15.2 +/- 1.9) days when recipients treated by CsA and (16.6 +/- 2.1) days by CTLA4Ig. Under the combination of CsA and CTLA4Ig, the mean survival time was significantly prolonged to (28.8 +/- 3.5) days (P < 0.05) with the lowest level of IL-2 in serum of recipients. CONCLUSION: The combined application of external CsA and CTLA4Ig inhibits rejection of allograft effectively, which can be a favorable therapy on composite tissue allo-transplantation.