Management of wounds with exposed bone structures using an induced-membrane followed by polymethyl methacrylate and second-stage skin grafting in the elderly with a 3-year follow-up.

The treatment of traumatic wounds with exposed bone or tendons is often challenging. An induced membrane (IM) is used to reconstruct bone defects, as it provides an effective and sufficient blood supply for bone and soft-tissue reconstruction. This study explored a novel two-stage strategy for wound management, consisting of initial wound coverage with polymethyl methacrylate (PMMA) and an autologous split-thickness skin graft under the IM. Fifty inpatients were enrolled from December 2016 to December 2019. Each patient underwent reconstruction according to a two-stage process. In the first stage, the defect area was thoroughly debrided, and the freshly treated wound was then covered using PMMA cement. After 4-6 weeks, during the second stage, the PMMA cement was removed to reveal an IM covering the exposed bone and tendon. An autologous split-thickness skin graft was then performed. Haematoxylin and eosin (H&E) staining and immunohistochemical analysis of vascular endothelial growth factor (VEGF), CD31 and CD34 were used to evaluate the IM and compare it with the normal periosteal membrane (PM). The psychological status and the Lower Extremity Function Scale (LEFS) as well as any complications were recorded at follow-up. We found that all skin grafts survived and evidenced no necrosis or infection. H&E staining revealed vascularised tissue in the IM, and immunohistochemistry showed a larger number of VEGF-, CD31- and CD34-positive cells in the IM than in the normal PM. The duration of healing in the group was 5.40 ± 1.32 months with a mean number of debridement procedures of 1.92 ± 0.60. There were two patients with reulceration in the group. The self-rating anxiety scale scores ranged from 35 to 60 (mean 48.02 ± 8.12). Postoperatively, the LEFS score was 50.10 ± 9.77. Finally, our strategy for the management of a non-healing wound in the lower extremities, consisting of an IM in combination with skin grafting, was effective, especially in cases in which bony structures were exposed in the elderly. The morbidity rate was low.

Co-culturing polarized M2 Thp-1-derived macrophages enhance stemness of lung adenocarcinoma A549 cells.

BACKGROUND: The tumor microenvironment (TME) is highly associated with cancer stem cells, and affects tumor initiation, progression, and metastasis. This study aimed to explore the underlying molecular mechanism of induction of A549 cancer cell stemness by THP-1-derived macrophages. METHOD: The Hedgehog inhibitor (Vismodegib), Notch inhibitor Gamma Secretase Inhibitor (GSI), and Signal Transducer and Activator of Transcription 3 (STAT3) inhibitor Cucurbitacin I (JSI-124) were added separately into the co-culture system of A549 cancer cell with THP-1-derived macrophages. Cell Counting Kit-8 (CCK-8) assay and the Cell-IQ continuous surveillance system were used to examine the cell growth and morphological changes of A549 cells. The messenger ribonucleic acid (mRNA) and protein expression levels of stem cell markers were respectively analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting, and the activity of Acetaldehyde dehydrogenase (ALDH) enzyme was assessed by flow cytometry analysis. Enzyme-linked immunosorbent assay (ELISA) and qRT-PCR assays were performed to evaluate the activation and differentiation of macrophages. RESULTS: Results showed that the proliferation and stemness of A549 cells were significantly enhanced by co-culturing with THP-1-derived macrophages. The expression levels of Transforming growth factor-ß (TGF-ß) and Interleukin-6 (IL-6) in macrophages were notably increased after co-culturing with A549 cells. Meanwhile, co-culturing with A549 cells induced the polarization of macrophages towards the M2 phenotype. Moreover, the inhibitors could reduce the proliferation and stemness of the co-culture system, and decrease the expression of TGF-ß and IL-6. CONCLUSIONS: These results suggested that co-culturing A549 cells with THP-1-derived macrophages could induce the stemness of A549 cells via multiple pro-tumorigenic pathways. Thus, inhibition of the interaction between macrophages and lung cancer stem cells may be a viable target for lung cancer treatment in the future.

5-FU blocks shuttling of HuR mediated by PKCδ in gastric cancer cells.

BACKGROUND: 5-fluorouracil (5-FU) is a common chemotherapy drug for gastric cancer. Human antigen R (HuR) is an RNA-binding protein that is also known as ELAV like RNA binding protein 1 (ELAVL1) regulates gene expression by binding to target genes 3' UTR region and is highly expressed in tumor tissues. However, the regulatory mechanisms of HuR in 5-FU mediated chemotherapy in stomach cancer are not well understood. In this study, we aimed to investigate 5-FU regulated PKCδ expression and translocation of HuR in SGC cell lines. METHODS: Using Cell viability assay to obtained IC50 doses when SGC cells were treated with 5-FU for 2 and 3 days. Western blot was used to detect the total protein content of HuR and PKCδ after treating with 20 mM 5-FU. Furthermore, after adding 20 mM 5-FU, knocking down PKCδ, HuR and using inhibitor Staurosporine respectively, the protein content of HuR in the cytoplasm were detected by Western blot and Immunofluorescence. MTT assay was used to verify the changes in cell proliferation after knockdown of HuR. RESULTS: Using Cell viability assay, we obtained IC50 doses of 77 and 20 mM, respectively, when SGC cells were treated with 5-FU for 2 and 3 days. The total protein content of HuR and PKCδ in SGC cells treated with 20 mM 5-FU did not change. However, Western blot and Immunofluorescence detected that the protein content of HuR in the cytoplasm decreased with 20 mM 5-FU treatment. Staurosporine inhibits the nucleation of HuR and is an inhibitor of PKC. Consistently, the expression of HuR in cytoplasm declined while knockdown of PKCδ. And the proliferation of SGC cells decreased after knocking down the HuR. CONCLUSIONS: Our results showed that 5-FU blocked shuttling of HuR from the nucleus to the cytoplasm in SGC cells through PKCδ phosphorylation.

Injuries due to foreign body ingestion and insertion in children: 10 years of experience at a single institution.

AIM: Foreign body (FB) injuries represent a severe public health problem during childhood. The aim of this study was to report our experience with patients with injuries due to FB ingestion and insertion who were treated surgically at our institution. METHODS: A total of 78 paediatric patients who were hospitalised for FB injuries were retrospectively reviewed. RESULTS: The series was composed of 27 males and 51 females, with a median age of 3.6 years. The cases included 35 cases of FB ingestion and 43 cases of FB insertion, including 40 cases with a vaginal insertion, 2 cases with a rectal insertion and 1 case with a urethra insertion. Intestinal perforation (n = 26) was a more common complication than intestinal obstruction (n = 9) in patients who had ingested a FB. The main clinical symptom was persistent vaginal discharge, followed by vaginal bleeding for patients with a vaginal FB insertion. Exploratory laparotomy was performed on 36 patients, while a laparoscopic approach was employed in 1 patient. Forty patients underwent hysteroscopy and one patient underwent cystotomy to remove the FB. All FBs were successfully removed. Of the 78 FBs recovered, 26 were food objects, while non-food objects were found in 52 patients. All patients recovered well, except one patient with an intestinal obstruction from adhesions that occurred approximately 1 month after discharge. CONCLUSIONS: Early recognition of FB injuries and appropriate management can significantly reduce complications. Surgical removal of a FB can be safe and effective, and relatively better outcomes can be achieved.

Chronic hyperglycemia regulates microglia polarization through ERK5.

Diabetic patients are prone to developing Alzheimer's disease (AD), in which microglia play a critical role. However, the direct effect of high glucose (HG) on microglia and the role of extracellular-signal-regulated kinase 5 (ERK5) signaling in this interaction have not been examined before. Here, these questions were addressed in microglia cultured in HG versus normal glucose (NG) conditions. Initially, HG induced microglial differentiation into the M2a phenotype with concomitant ERK5 activation. However, longer exposure to HG further induced differentiation of microglia into the M2b-like phenotype, followed by the M1-like subtype, concomitant with a gradual loss of ERK5 activation. BIX021895, a specific inhibitor of ERK5 activation, prevented M2a- differentiation of microglia, but induced earlier M2b-like polarization followed by M1-like polarization. Transfection of microglia with a sustained activated form of MEK5 (MEK5DD) prolonged the duration of the M2a phenotype, and prevented later differentiation into the M2b/M1 subtype. Conditioned media from the M2a-polarized microglia reduced neuronal cell apoptosis in hypoxic condition, while media from M2b-like or M1-like microglia enhanced apoptosis. Together, our data suggest that chronic hyperglycemia may induce a gradual alteration of microglia polarization into an increasingly proinflammatory subtype, which could be suppressed by sustained activation of ERK5 signaling.