STAT3-induced lncRNA GNAS-AS1 accelerates keloid formation by mediating the miR-196a-5p/CXCL12/STAT3 axis in a feedback loop.

Keloids are pathological scar tissue resulting from skin trauma or spontaneous formation, often accompanied by itching and pain. Although GNAS antisense RNA 1 (GNAS-AS1) shows abnormal upregulation in keloids, the underlying molecular mechanism is unclear. The levels of genes and proteins in clinical tissues from patients with keloids and human keloid fibroblasts (HKFs) were measured using quantitative reverse transcription PCR, western blot and enzyme-linked immunosorbent assay. The features of HKFs, including proliferation and migration, were evaluated using cell counting kit 8 and a wound healing assay. The colocalization of GNAS-AS1 and miR-196a-5p in HKFs was measured using fluorescence in situ hybridization. The relationships among GNAS-AS1, miR-196a-5p and C-X-C motif chemokine ligand 12 (CXCL12) in samples from patients with keloids were analysed by Pearson correlation analysis. Gene interactions were validated by chromatin immunoprecipitation and luciferase reporter assays. GNAS-AS1 and CXCL12 expression were upregulated and miR-196a-5p expression was downregulated in clinical tissues from patients with keloids. GNAS-AS1 knockdown inhibited proliferation, migration, and extracellular matrix (ECM) accumulation of HKFs, all of which were reversed by miR-196a-5p downregulation. Signal transducer and activator of transcription 3 (STAT3) induced GNAS-AS1 transcription through GNAS-AS1 promoter interaction, and niclosamide, a STAT3 inhibitor, decreased GNAS-AS1 expression. GNAS-AS1 positively regulated CXCL12 by sponging miR-196-5p. Furthermore, CXCL12 knockdown restrained STAT3 phosphorylation in HKFs. Our findings revealed a feedback loop of STAT3/GNAS-AS1/miR-196a-5p/CXCL12/STAT3 that promoted HKF proliferation, migration and ECM accumulation and affected keloid progression.

Auricular Reconstruction in Adult Patients With Unprepared Congenital Microtia: A Single Institution's Experience.

METHODS: From January 2015 to January 2021, auricular reconstruction was performed in 38 adult patients (39 ears) of congenital microtia based on autologous costal cartilage. The whole procedure was divided into 2 stages: stage I, the individualized framework carved with autologous costal cartilage was inserted into subcutaneous pocket in the mastoid region; then, the earlobe was transposed backward; and stage II, ear elevation, harvesting the retroauricular fascial flap to cover the support scaffold and closing the defect with free skin graft, was performed. RESULTS: All patients successfully underwent ear reconstruction. The follow-up time ranged from 3 months to 3 years. Infection occurred in 1 patient. The ear frameworks were partially broken at the helix in 4 cases. Retroauricular graft skin survival was poor in 1 patient. Retroauricular hypertrophic scars occurred in 2 cases. Bad projection of the reconstructed ear occurred in 1 case. Totally 38 patients were satisfied with the results. CONCLUSIONS: According to the physiological characteristics of the costal cartilage and skin soft tissues of adult patients, improvements are made to details based on the Nagata's method, so that the adult patients with microtia can obtain satisfactory surgical results.

Simultaneous measurements of thin film thickness using total internal reflection fluorescence microscopy and disjoining pressure using Scheludko cell.

This work describes a method for measuring the thin film thickness using total internal reflection fluorescence microscopy, with the use of evanescent wave illumination. The thin liquid film was formed in a hole drilled at the center of a porous plate, which is used for measurement of the disjoining pressure by using the Scheludko cell method. The aim of simultaneous and in situ measurements of thin film thickness and disjoining pressure is to obtain the relationship between them, which is critical for explicitly depicting the thin film profile that determines the interfacial mass and heat fluxes in the thin film region near the triple line. This method can overcome the drawbacks of the optical methods that are insufficient for measuring the thickness of a thin film with curvature. The influence of structural forces formed by tracer nanoparticles seeded in the thin liquid film on the relationship was analyzed. The obtained expression for disjoining pressure vs thin film thickness provides a basis for analyzing the formation, evolution, and stability of the thin liquid film, which is the dominant mechanism of controlling the mesoscopic structure in many transport processes.

Interleukin 17 (IL-17)-Induced Mesenchymal Stem Cells Prolong the Survival of Allogeneic Skin Grafts.

BACKGROUND Mesenchymal stem cells (MSCs) have the potential of self-renewal and multi-differentiation and have a wide application prospect in organ transplantation for the effect of inducing immune tolerance. It has found that interleukin 17 (IL-17) could enhance the inhibition effect of MSCs on T cell proliferation and increase the immunosuppressive effect of MSCs. In this study, we aimed to investigate the effect of IL-17-induced MSCs on allograft survival time after transplantation. MATERIAL AND METHODS BMSCs were characterized by differential staining. The allogenic skin transplantations were performed and the BMSCs pre-treated by IL-17 were injected. To assess the immunosuppressive function of IL-17-induced BMSCs, the morphology of the grafts, the homing ability of the BMSCs, and the survival time of the grafts were analyzed. RESULTS BMSCs from BALB/c have multidirectional differentiation potential to differentiate into osteogenic, chondrogenic, and adipogenic lineage cells. IL-17-induced BMSCs prolonged the survival time of allogeneic skin grafts dramatically. We found that there were more labeled MSCs in the skin grafts, and the Treg subpopulations percentage, IL-10, and TGF-ß were significantly increased, while the IFN-γ level was decreased compared to the control group and MSCs group. In conclusion, IL-17 can enhance the homing ability of MSCs and regulate the immunosuppressive function of MSC. CONCLUSIONS Our data demonstrate that IL-17 plays the crucial role in MSC homing behaviors and promotes immunosuppression of MSCs during transplantation procedures, suggesting that IL-17-pre-treated MSCs have potential to prolong graft survival and reduce transplant rejection.

Modified 2-Stage Method for Auricular Reconstruction.

BACKGROUND: Auricular reconstruction is one of the most challenging procedures in the field of plastic surgery. The aims of this study were to apply the modified 2-stage method to perform auricular reconstruction and to summarize the clinical experience in the past 10 years. METHODS: Auricular reconstruction was performed in 243 patients (total 254 ears) of congenital microtia using autologous costal cartilage. The whole procedure is divided into 2 stages. Stage I: the individualized framework fabricated with autologous costal cartilage is inserted into subcutaneous pocket in the mastoid region, and then the earlobe was transposed backward to connect with the lower part of the framework. The second-stage surgery mainly includes the following main contents: ear elevation, fixing the bracket behind the reconstructed ear framework, harvesting the retroauricular fascial flap to cover the bracket, and closing the defect with free skin grafts. RESULTS: A total of 243 patients (254 ears) of congenital microtia underwent ear reconstruction. The follow-up time ranged from 6 months to 4 years; 220 patients were satisfied with the results. Surgery-related complications such as infection, partial skin graft necrosis, flap necrosis, bad projection of the constructed auricle, and extrusion of cartilage occurred in 24 cases, and hypertrophic scars occurred in 16 patients. CONCLUSIONS: This modified 2-stage method for auricle reconstruction can receive acceptable results and fewer complications; furthermore, it is relatively simple and easy to master. The 10-year experience validates that this modified method is an ideal method in auricular reconstruction.

MiR-139-5p inhibits the tumorigenesis and progression of oral squamous carcinoma cells by targeting HOXA9.

Our study sought to clarify the effects of microRNA-139-5p (miR-139-5p) in the tumorigenesis and progression of oral squamous cell carcinoma (OSCC) by regulating HOXA9. MiR-139-5p and HOXA9 expression in OSCC tissues, tumour adjacent tissues, OSCC cells and normal cells were tested by qRT-PCR. SAS and CAL-27 cell lines were selected in among four OSCC cell lines and then transfected with miR-139-5p mimics, pEGFP-HOXA9 and cotransfected with miR-139-5p mimics + pEGFP-HOXA9. We used MTT, colony formation, transwell and wound healing assays to analyse cell viability, proliferation, invasion and migration. The target relationship between miR-139-5p and HOXA9 was verified by luciferase reporter assay and Western blot, respectively. MiR-139-5p was down-regulated, whereas HOXA9 was up-regulated in OSCC tissues and cells. The proliferation, invasion and migration ability of SAS and CAL-27 cells in miR-139-5p mimics group were significantly weaker than those in the control group and the miR-NC group (P < 0.01). MiR-139-5p can negatively regulate HOXA9. The proliferation, invasion and migration of SAS and CAL-27 cells in the miR-139-5p mimics + pEGFP-HOXA9 group were not significantly different from those in the blank control and negative control groups (P > 0.05). Our results indicated that miR-139-5p could directly inhibit HOXA9, which might be a potential mechanism in inhibiting the proliferation, invasiveness and migration of OSCC cells.

MiR-376c-3p regulates the proliferation, invasion, migration, cell cycle and apoptosis of human oral squamous cancer cells by suppressing HOXB7.

PURPOSE: To test the influence of miR-376c-3p on the proliferation, invasion, migration, cell cycle and apoptosis of human oral squamous cancer cells (OSCC) and the relevant mechanism. METHODS: We applied qRT-PCR and Western blot to compare the expression level of miR-376c-3p and HOXB7 in SCC-4, SCC-9, SCC-15, SCC-25 OSCC cell lines and 49 paired OSCC and normal oral epithelial tissue specimens were included in our present study. Also we analyzed the relative relationship of expression level between miR-376c-3p and HOXB7 in cancer tissues. Luciferase assay was used to confirm the target relationship between miR-376c-3p and HOXB7. Besides, MTT, Transwell, wound healing, colony formation and flow cytometer experiments were applied to evaluate the proliferation, cell viability, apoptosis, invasion and migration of transfected OSCC. RESULTS: MiR-376c-3p was down-regulated while HOXB7 was up-regulated in OSCC tissues and cells than the normal ones. MiR-376c-3p directly targeted HOXB7 and reduced the expression of HOXB7. Overexpression of miR-376c-3p attenuated proliferation of SCC-9, SCC-15, SCC-24 and SCC-25 cells. Moreover, miR-376c-3p suppressed proliferation, viability, migration and invasion and induced G1/G0 arrest and cell apoptosis of SCC-25 cells. Besides, overexpression of HOXB7 efficiently abrogates these influences caused by overexpression of miR-376c-3p. CONCLUSION: MiR-376c-3p suppresses the fission, proliferation, migration and invasion and induces cell apoptosis of OSCC via targeting HOXB7.

Immune Tolerance of Mesenchymal Stem Cells and Induction of Skin Allograft Tolerance.

BACKGROUND: Mesenchymal stem cells not only possess reparative properties, but also have immunomodulatory effect. Owing to the properties, they have been proposed to be hopeful candidates for cell therapy in the process of organ transplantation. In the preclinical researches, it shows that MSCs is capable of prolonging graft survival and inducing tolerance in some cases. Various mechanisms of immune tolerance were reported before, such as tolerogenic dendritic cells, induction of apoptosis, regulatory T cells, mixed chimerism, soluble factors and anergy. Furthermore, the induction of immune tolerance may be influenced by the dose, route and optimal timing of MSCs administration. Allograft of skin can provisionally restore the function of skin barrier and offer a good environment to expand micro-skin auto-graft; however, at the same time, it can cause strong immune rejection, which leads to the failure of skin graft. OBJECTIVE: The review aims at analyzing the mechanisms of immunosuppression mediated by MSCs, and the induction of tolerance of skin graft by MSCs. CONCLUSION: Mesenchymal stem cells are hopeful candidates for cell therapy in the process of organ transplantation, and can induce immune tolerance of skin graft to some extent.

Tissue expansion in the treatment of giant congenital melanocytic nevi of the upper extremity.

The aim of our study was to use tissue expansion for the treatment of giant congenital melanocytic nevi of the upper extremity and examine potential advantages over traditional techniques.There were 3 stages in the treatment of giant congenital melanocytic nevi of the upper extremities using tissue expansion: first, the expander was inserted into the subcutaneous pocket; second, the expander was removed, lesions were excised, and the wound of the upper extremity was placed into the pocket to delay healing; third, the residual lesion was excised and the pedicle was removed. The pedicle flap was then unfolded to resurface the wound.During the period between June 2007 and December 2015, there were 11 patients with giant congenital melanocytic nevi of the upper extremities who underwent reconstruction at our department with skin expansion. Few complications were noted in each stage of treatment. The functional and aesthetic results were observed and discussed in this study.Optimal aesthetic and functional results were obtained using tissue expansion to reconstruct the upper extremities due to the giant congenital melanocytic nevi.

PRMT1 regulates tumor growth and metastasis of human melanoma via targeting ALCAM.

Overexpression of protein arginine methyltransferases (PRMTs) is associated with various types of cancer. The present study aimed to determine the expression level of PRMT1 in human melanoma and investigate its biological function. The clinical significance of PRMT1 was determined by screening the Oncomine database, and the increased expression of PRMT in melanoma was confirmed by western blot analysis. Furthermore, the current study demonstrated that PRMT1 was overexpressed in melanoma cell lines compared with human immortalized keratinocytes and PIG1 immortalized human melanocytes. Silencing PRMT1 in A375 and Hs294T cells significantly suppressed tumor growth and metastatic ability of the melanoma cell line compared with the negative control. These changes were in accordance with the upregulation of the cadherin 1 level and downregulation of several metastatic­associated genes determined by a quantitative polymerase chain reaction array. Liquid chromatography­mass spectrometry demonstrated that activated leukocyte cell adhesion molecule (ALCAM) may be a direct target of PRMT1, and the interaction was confirmed by co­immunoprecipitation. Compared with negative controls, the protein level of ALCAM was decreased following the silencing of PRMT1, and re­expression of ALCAM in A375/shPRMT1 or Hs294T/shPRMT1 cells using an expression vector restored the colony formation and metastatic ability of the cells. In conclusion, the current results indicated that PRMT1 is overexpressed in human melanoma, and may regulate tumor growth and metastasis via targeting ALCAM.

Reconstruction of Philtrum Using Partial Splitting and Folding of Orbicularis Oris Muscle in Secondary Unilateral Cleft Lip.

BACKGROUND: The reconstruction of the philtrum during unilateral cleft lip repair remains a surgical challenge. The authors introduce a modified surgical technique for the reconstruction of the philtrum that allows for both the deepening of the philtral dimple and augmentation of the philtral ridge. METHODS: Between January of 2011 and June of 2012, 37 patients, including 22 male and 15 female patients (mean age, 25.6 years), underwent secondary unilateral cleft lip repair. The philtrum was reconstructed by the partial splitting and folding of the orbicularis oris muscle. The aesthetic outcome of the philtrum and the scar after the operation was scored to evaluate the effectiveness of the procedure. RESULTS: The patients were evaluated for the appearance of the philtrum and scar at 12 to 18 months (mean, 14.6 months). Good symmetry of the bilateral philtral ridges was achieved in 35 of the 37 patients, with the postoperative scores for aesthetic outcome being 2 to 4. The postoperative appearance of the scar in almost all cases was also significantly better than that before the operation. Only two patients showed widening of the philtral ridges and less prominent philtral ridges on the reconstructed side than on the normal side. CONCLUSIONS: The authors presented a new technique for the repair of the philtrum in patients with secondary unilateral cleft lip; the technique afforded good aesthetic outcomes in almost all cases.

MicroRNA 26a prolongs skin allograft survival and promotes regulatory T cell expansion in mice.

MicroRNA 26a (Mir-26a) has been reported to play functions in cellular differentiation, cell growth, cell apoptosis, and metastasis. However, the role of Mir-26a in transplant rejection has never been investigated. Full-thickness skin grafts 1-2 cm in diameter were obtained from the tail-skin CBA/J donor mice and transplanted onto the back of wild-type C57Bl/6 recipient mice. Vectors encoding pre-Mir-26a (LV-26a) and an empty lentiviral vector (LV-Con) delivered approximately 2 × 10(7) transforming units of recombinant lentivirus were injected to mice once through the tail vein. Mir-26a overexpression results in prolonged skin allograft survival (MST = 9.5 days in LV-Con mice; MST = 22 days in LV-26a mice. P < 0.01) and promoted regulatory T cells (Tregs) expansion. The prolonged skin allograft survival induced by LV-26a was abrogated by depletion of Tregs with anti-CD25 antibodies. Mir-26a significantly promoted IL-10 expression and suppressed the expression of IL-6, IL-17, and IFN-γ. Furthermore, IL-6 overexpression led to complete suppression of the Mir-26a-induced upregulation of Foxp3. The prolonged allograft survival induced by LV-Mir-26a was also completely abrogated by IL-6 overexpression. In conclusion, Mir-26a prolongs skin allograft survival and promotes Tregs expansion in part through inhibition of IL-6 expression.

[Auricular reconstruction for microtia with modified Nagata method].

OBJECTIVE: To investigate a method for total reconstruction of auricle. METHODS: 90 patients (101 ears) with congenital microtia underwent two-stage operations for auricular reconstruction. The first stage involved fabrication and grafting of autologous costal cartilage, removing the remnant ear cartilage, embedding the framework into local flap of the mastoid region, transferring the remnant ear lobule flap to link to the inferior framework. The second stage was creating an auriculocephalic sulcus. The reconstruction was performed 4 - 12 months after the first surgery. Skin incision was made 5 mm lateral side of the posterior margin of the auricle. The ear framework carrying a thick ear fascia was separated from the side of the head, the frames of the costal cartilage banked at the first operation were harvested, shaved and transplanted to the posterior wall of the concha with sutures; adjust stand position and angle, so that made the ear shape, position, axis, close to the healthy ear, and auriculocephalic angle was slightly larger than the contralateral ear. Two random flap was designed with superior on the root of the helix and in the inferior-posterior direction of the inferior mastoid area, two flapes were elevated and transplanted to posterior auricular sulcus to cover the grafted cartilage. Skin graft was performed in the remaining raw surface. RESULTS: A total of 90 patients were operated, all of 101 constructed ears achieved satisfied or near satisfied shapes. Five cases of partial skin flap necrosis were caused by pedicle impairment. Exposure of cartilage framework happened in two cases. The auriculocephalic sulcus of four cases diminished after the second stage operation. Three month to two-year follow-up of 67 patients showed that the reconstructed ears were satisfied with the results, including good shapes and steady auriculocephalic angles. CONCLUSIONS: The method is a simple, safe and reliable method for total aural reconstruction.

[Reconstruction of segmental ear helix defect with post-auricular skin flap].

OBJECTIVE: To explore the method of repairing segmental ear helix defect. METHODS: Twenty-one patients with segmental ear helix defect were repaired with post-auricular skin flap. In the first stage operation, ear helix defect was assessed, including the anterior and posterior area defect. According to the defect, post-auricular skin flap was designed and transplanted to repair the defect. Six weeks later, the pedicle of the post-auricular skin flap was cut off, elevated, and folded to form the helix. The secondary defect was directly sutured or repaired with skin graft. RESULTS: Twenty-one patients were treated with this method. In two to 12 months follow-up, all flaps survived and reconstructed ear helices were in good shape. The reconstructed ears were in symmetry to the healthy ones. CONCLUSION: The method is safe and effective for the correction of segmental ear helix defect.

Burden, anxiety and depression in caregivers of veterans with dementia in Beijing.

Dementia is the most common neurodegenerative disease in the elderly. The number of Chinese dementia patients reached 5 million in 2001 and will grow rapidly by 314-336% in 2040. Caring for patients with dementia is a stressful experience, which can cause the physical and mental problems in caregivers. According to the protocol, we evaluated the burden, depression and anxiety of caregivers by Clinical Dementia Rating (CDR) Scale, Caregiver Burden Inventory (CBI), Self-Rating Depression Scale (SDS) and Self-Rating Anxiety Scale (SAS). 90 caregivers of dementia patients and 90 caregivers of nondementia patients with other chronic diseases from 24 military communities in Beijing were enrolled in this investigation. The scores of burden, depression and anxiety increased from nondementia to severe dementia groups. There were associations between patient's CDR score and CBI score, as well as the average daily care time and CBI score. In addition, the scores of SDS and SAS were both positive correlated to CBI score. Comparing to general community, the caregivers of dementia veterans had lower scores in SDS and SAS. Therefore, we should pay more attentions to the caregivers of severe dementia patients. Reduction of daily care time is a possible way to releasing the burden of caregivers. Better general supports are beneficial to reducing depression and anxiety when caregivers confront the heavy burden.

[Modified two-stage surgery for total auriculoplasty with autogenous rib cartilage].

OBJECTIVE: To introduce a modified surgery for total auriculoplasty and the experience in one hundred and forty-six cases (155 ears). METHODS: The procedure was a two-stage operation. The first stage involved fabrication and grafting of a costal cartilage framework. A U-shaped skin incision was made on the posterior edge of the lobule and the remnant ear cartilage was removed completely. The area for the insertion of the cartilage framework was undermined. Skin flaps were sutured after insertion of the cartilage framework. The second-stage surgery was usually performed six months after the first-stage operation. The reconstructed auricle was elevated, and a costal cartilage block was fixed to the posterior part of the auricle. A temporoparietal fascia flap was then used to cover the costal cartilage block. Finally, the posterior aspect of the projected auricle was covered with a spit-thickness skin graft. RESULTS: The incisions healed in one hundred and forty-one patients (150 ears) after the first stage operation. Partial necrosis of the postauricular flap was observed in five cases (5 ears) after the first stage operation, but no exposure or absorption of the cartilage took place. The skin grafts survived in one hundred and thirty-nine cases (147 ears) after the second-stage surgery. Partial necrosis of the skin graft was observed in seven cases (8 ears), but healed after one-week of dressing changes. Ninety-four cases (97 ears) were followed up, but fifty-two cases (58 ears) were lost to follow up. The follow-up at six months to two years showed satisfactory contour and projection of the constructed ears. CONCLUSION: This two-stage surgery is simple and ideal for auricloplasty with few complications.

[Neuropsychiatric symptoms in dementia and elderly people in the community: results from the Beijing Dementia Cooperative Study].

OBJECTIVE: To determine the prevalence of neuropsychiatric symptoms in dementia and normal elderly people living in the Chinese community of Beijing. METHODS: A cross-sectional study derived from the Beijing Dementia Cooperative Study was carried out a population survey was carried out on a total of 1540 participants aged 65 years and older living in Beijing city and rural areas. All the individuals and 373 demented elderly people completed a series of neuropsychological examination and the Neuropsychiatric Inventory (NPI). RESULTS: Among the dementia participants, 49.33% had exhibited neuropsychiatric symptoms (35.66% rated as clinically significant), in which 80.4% reported 2 or more disturbances, with depression (23.86%), apathy (21.72%) and anxiety (20.38%) being most common. Of the 1540 normal individuals, 18.25% of them exhibited neuropsychiatric symptoms (6.49% rated as clinically significant), in which 53% reported 2 or more disturbances, with sleepless (10%), depression (8.9%) and anxiety (6.97%) being the most common. CONCLUSION: To our knowledge, this was the first multi-center study on neuropsychiatric disturbances in dementia and cognitive normal elderly people. Neuropsychiatric symptoms occurred mainly in persons with dementia and of clinical severity. Though the neuropsychiatric disturbances reported in cognitive normal individuals were lower and less serious compared to dementia, they should not be neglected. These finding suggested that a screening programme focusing on identifying these symptoms should be included in the physician's diagnostic tools for dementia.