Reconstruction of Squamous Cell Carcinoma on Oral Commissure With Hatchet-Shaped Flap.

The mouth is a unique and prominent element of the lower face. Given the complex anatomy, aesthetic appearance, and function of the oral commissure, its reconstruction due to various causes presents a significant challenge for surgeons. Squamous cell carcinoma (SCC) of the lip is the most common type of oral cancer, accounting for approximately 25% to 30% of all oral cancers. Wide excision is the treatment of choice, and the prognosis is generally favorable. We encountered a case of SCC of the right oral commissure in a 69-year-old man. We designed a hatchet-shaped flap to minimize anatomical disruption and, as a result, achieved satisfactory outcomes in terms of both functionality and aesthetics.

Reconstruction of Rapidly Growing Merkel Cell Carcinoma of the Cheek With Multidisciplinary Cooperation.

Merkel cell carcinoma (MCC) is a rare and very aggressive skin cancer. An 83-year-old female presented with a 1.5 cm-sized non-tender mass on her left cheek and was diagnosed with MCC. The margin of MCC was well-defined and there was no cervical node metastasis on pre-operative computed tomography. Three weeks after the first visit, the mass rapidly increased in size. We checked the magnetic resonance imaging, a rapid-growing 2.5 cm sized nodular region and metastatic cervical lymph node were found. We performed wide excision of the MCC and neck lymph need dissection with multidisciplinary cooperation. The soft tissue defect was about 6.0×5.0 cm 2 in size and reconstructed with radial forearm free flap. On permanent biopsy, the size of MCC was 3.0×2.3 cm 2 . There was no recurrence of MCC with radiation therapy during an 18-month follow-up. We experienced an older patient with a rapid - growing MCC and cervical lymph node metastasis in a brief time. With our experience, we discuss the evaluation and treatment plan of the rapid-growing MCC for good results.

Teaching Models for Correct Rhombic Flaps.

The aim of this study was to introduce teaching models for correct rhombic flaps. For the line of maximal extensibility (LME) and flap design, surgical fabric (model 1), scored corrugated cardboard (model 2), and scored polyethylene sheet (model 3) were used. For choosing the correct flaps, a silicone face (model 4) was used. Seven participants in the Plastic Surgery Department were recruited for the workshop. In models 1 to 3, a 2-cm diameter circle and relaxed skin tension line were indicated. Participants were requested to design Limberg flaps. Each flap was elevated, transposed, and fixed with sutures (model 1) or cellophane tape (models 2 and 3). In model 4, a 1-cm diameter circle was indicated on the cheek. Participants were requested to design correct Limberg flaps. Although participants were not provided an article describing how to create correct Limberg flaps, they eventually created correct flaps through trial and error. Participants drew 2 parallel lines tangential to the defect and following the LME, perpendicular to the relaxed skin tension lines, which are the same as the scoring marks. They then drew 2 other sides of 2 possible parallelograms by tilting them medially and laterally with angles of 60 and 120 degrees each. Thus, 4 possible Limberg flaps to close the defect were drawn. Among the 8 possible flaps, 4 flaps that did not follow the LME were eliminated. Scored polyethylene sheet had the best extensibility and least distortion among the 3 models. Through this workshop, participants learned to design rhombic flaps correctly, using 2 parallel LMEs.

Anhydrous Alum Inhibits α-MSH-Induced Melanogenesis by Down-Regulating MITF via Dual Modulation of CREB and ERK.

Melanogenesis, the intricate process of melanin synthesis, is central to skin pigmentation and photoprotection and is regulated by various signaling pathways and transcription factors. To develop potential skin-whitening agents, we used B16F1 melanoma cells to investigate the inhibitory effects of anhydrous alum on melanogenesis and its underlying molecular mechanisms. Anhydrous alum (KAl(SO4)2) with high purity (>99%), which is generated through the heat-treatment of hydrated alum (KAl(SO4)2·12H2O) at 400 °C, potentiates a significant reduction in melanin content without cytotoxicity. Anhydrous alum downregulates the master regulator of melanogenesis, microphthalmia-associated transcription factor (MITF), which targets key genes involved in melanogenesis, thereby inhibiting α-melanocyte-stimulating hormone (α-MSH)-induced melanogenesis. Phosphorylation of the cAMP response element-binding protein, which acts as a co-activator of MITF gene expression, is attenuated by anhydrous alum, resulting in compromised MITF transcription. Notably, anhydrous alum promoted extracellular signal-regulated kinase phosphorylation, leading to the impaired nuclear localization of MITF. Overall, these results demonstrated the generation and mode of action of anhydrous alum in B16F1 cells, which constitutes a promising option for cosmetic or therapeutic use.

α-ketoisocaproic acid promotes ER stress through impairment of autophagy, thereby provoking lipid accumulation and insulin resistance in murine preadipocytes.

α-ketoisocaproic acid (AKA), a metabolite of l-leucine, is one of the branched-chain amino acids (BCAAs) involved in energy metabolism. However, the effects of AKA on lipid metabolism, insulin signaling, and related mechanisms in preadipocytes have not been reported. Herein, we investigated the impacts of AKA on lipid accumulation in 3T3-L1 murine preadipocytes. Treatment with AKA for 4 days enhanced lipid accumulation and expression of lipogenic proteins, such as cleaved sterol-regulatory element-binding proteins (SREBP1) and stearoyl-CoA desaturase-1 (SCD1) in 3T3-L1 cells. Increased endoplasmic reticulum (ER) stress markers, such as phosphorylation of eukaryotic initiation factor 2 (eIF2) and CHOP, were observed in the presence of AKA. AKA treatment increased mTOR phosphorylation and reducing autophagy markers, such as LC3 conversion and degradation of p62. Treatment with rapamycin, an mTOR inhibitor, reduced the effects of AKA on ER stress and lipogenesis in 3T3-L1 preadipocytes. The present study demonstrates that AKA increases ER stress by impairing mTOR/autophagy signaling, thus promoting lipid accumulation and insulin resistance in preadipocytes. In particular, if AKA is taken together with substances that can suppress ER stress, more effective skeletal muscle gain will be possible.

Photothermal therapy using gold nanoparticles for acne in Asian patients: A preliminary study.

Acne is a common skin disease that occurs in pilosebaceous units and is often prevalent in adolescence. There are many acne treatments, but they are associated with side effects, such as antibiotic resistance, teratogenicity, and irritation. Therefore, it is necessary to develop a more effective and safe alternative treatment for managing acne in patients of all ages. This study aimed to confirm the effect of gold photothermal therapy for acne. About 12 patients who visited the dermatologic clinic with moderate to severe acne vulgaris were included in the study, regardless of age or sex. All patients received three successive treatments at 1- to 2-week intervals with a photopneumatic device after applying the contents of a gold nanoparticle ample to the skin. Changes in the number of papules, pustules, and comedones before and after treatment, along with the overall improvement, were assessed. In four patients, a biopsy was taken before and 1 month after the last treatment. Significant reductions in acne lesions were observed after the use of gold photothermal therapy (papules, P = .001; pustules, P < .001; and comedones, P = .001). As noted in the Physician Global Assessment, the patients showed an average improvement of more than 50% in their condition. In the histopathological findings, a decrease in inflammatory cell infiltration and fibrotic changes of the dermis were observed after gold photothermal therapy. Gold photothermal therapy showed significant clinical and histological improvements in acne vulgaris in Asians without serious adverse effects.

Dasatinib, a second-generation tyrosine kinase inhibitor, induces melanogenesis via ERK-CREB-MITF-tyrosinase signaling in normal human melanocytes.

Dasatinib, a second-generation tyrosine kinase inhibitor, is indicated for the therapy of imatinib-resistant leukemia and also for the treatment of solid cancers. Here, we report a novel effect of dasatinib of inducing differentiation in normal human melanocytes. Treatment with dasatinib significantly increased the melanin content and tyrosinase activity through the up-regulation of MITF and tyrosinase expressions. Consistently, dasatinib had clear stimulatory action in the pigmentation of ex vivo cultured skin. The molecular mechanism underlying the melanogenic effect of dasatinib was associated with the ERK-dependent phosphorylation of CREB. The ERK inhibitor PD98059 not only inhibited the phosphorylation of CREB but also abrogated dasatinib-induced melanocyte differentiation. These results demonstrate for the first time the capacity of dasatinib to induce differentiation in normal human melanocytes depending on the activation of ERK-CREB-MITF-tyrosinase signaling cascades.

Abrogation of B-RafV600E induced senescence by FoxM1 expression.

B-RafV600E oncogene mutation occurs in various cancers and is associated with tumor initiation. However, genetic modification of B-RafV600E in cells induces MAPK activation and results in oncogene-induced senescence. Overcoming the oncogene-induced senescence by B-RafV600E requires activation of another oncogene pathway, such as AKT signaling. In the present study, we explored the factors involved in overcoming the senescence program in cells activated by B-RafV600E and AKT signaling. B-RafV600E activation caused a feedback inhibition of AKT phosphorylation and resulted in downregulation of FoxM1, one of the AKT downstream components. AKT activation by PTEN downregulation induced FoxM1 expression, and co-expression of B-RafV600E and FoxM1 overcame the cellular senescence. These observations suggested that FoxM1 is critical downstream gene of AKT and functions to overcome B-RafV600E-induced senescence.

Senescent fibroblasts in melasma pathophysiology.

It has been proposed that melasma is a photoageing skin disorder. The photoaged fibroblasts have been suggested as an important source of melanogenic factors which are involved in the regulation of pigmentation. To investigate whether melasma includes senescent cells, lesional and perilesional normal skin from 38 melasma patients was assessed using a cell senescence marker, p16INK4A . The results showed that lesional dermal skin had more p16INK4A -positive senescent cells than perilesional skin. The impact of senescent fibroblasts was further investigated in a pilot study using radiofrequency (RF) intervention for melasma. It showed that the RF therapy decreased the number of senescent cells with increased expression of procollagen-1, which were associated with reduced epidermal pigmentation. This leads us to the speculation that senescent fibroblasts may contribute to drive melasma and might be considered as a potential therapeutic target.

Comparative effects of various absorbable threads in a rat model.

BACKGROUND: Conventional procedures including botulinum toxin and filler injections have their limitations in improving deep wrinkles and decreasing tissue laxity, and possess the propensity for vascular accidents. Absorbable thread is a recently commercialized field, but there is little evidence on comparative superiority. OBJECTIVES: We observed the effects of polydiaxanone (PDO) threads with different number of strands in relation to collagen production and histopathology in a rat model. MATERIALS AND METHODS: Dorsal skin of rat was divided into five different compartments and four different PDO threads and monofilament poly-lactic acid (PLA) thread were inserted. Tissue samples were obtained at week 1, 2, and 12 after the procedure for histopathologic review and real-time PCR for quantification of collagen. RESULTS: Multiple PDO filaments produced more collagen at 2 weeks. Single-stranded PLA thread insertion resulted in more Col1α1 levels than the double PDO thread and also showed the most Col1α3 production at week 2. The amount of collagen showed a sharp decline at week 12. Histologic evaluation showed retained threads surrounded by fibrous capsule-like structure at week 12. CONCLUSION: We were able to observe more collagen production in multiple stranded PDO threads compared to a single strand and that increasing number of threads leads to more collagen synthesis.

Paracrine crosstalk between endothelial cells and melanocytes through clusterin to inhibit pigmentation.

Cutaneous vasculature systems play a role in regulating skin pigmentation. We analysed RNA sequencing data to identify novel antimelanogenic factors secreted from endothelial cells and found that one of the secreted factors, clusterin, is highly expressed by HDMECs. To investigate the paracrine effect of clusterin from HDMECs on the regulation of melanogenesis, HDMECs were infected with clusterin or sh-clusterin lentivirus and the HDMEC-derived conditioned media were used to treat normal human melanocytes. It was found that HDMEC-derived clusterin inhibits melanogenesis through MITF/tyrosinase downregulation. The findings here suggest that HDMECs secrete copious amounts of clusterin and that the clusterin is a factor contributing to the inhibitory effect of endothelial cells on melanogenesis via paracrine crosstalk between endothelial cells and melanocytes.

Origin of myofibroblasts in the fibrotic liver in mice.

Hepatic myofibroblasts are activated in response to chronic liver injury of any etiology to produce a fibrous scar. Despite extensive studies, the origin of myofibroblasts in different types of fibrotic liver diseases is unresolved. To identify distinct populations of myofibroblasts and quantify their contribution to hepatic fibrosis of two different etiologies, collagen-α1(I)-GFP mice were subjected to hepatotoxic (carbon tetrachloride; CCl4) or cholestatic (bile duct ligation; BDL) liver injury. All myofibroblasts were purified by flow cytometry of GFP(+) cells and then different subsets identified by phenotyping. Liver resident activated hepatic stellate cells (aHSCs) and activated portal fibroblasts (aPFs) are the major source (>95%) of fibrogenic myofibroblasts in these models of liver fibrosis in mice. As previously reported using other methodologies, hepatic stellate cells (HSCs) are the major source of myofibroblasts (>87%) in CCl4 liver injury. However, aPFs are a major source of myofibroblasts in cholestatic liver injury, contributing >70% of myofibroblasts at the onset of injury (5 d BDL). The relative contribution of aPFs decreases with progressive injury, as HSCs become activated and contribute to the myofibroblast population (14 and 20 d BDL). Unlike aHSCs, aPFs respond to stimulation with taurocholic acid and IL-25 by induction of collagen-α1(I) and IL-13, respectively. Furthermore, BDL-activated PFs express high levels of collagen type I and provide stimulatory signals to HSCs. Gene expression analysis identified several novel markers of aPFs, including a mesothelial-specific marker mesothelin. PFs may play a critical role in the pathogenesis of cholestatic liver fibrosis and, therefore, serve as an attractive target for antifibrotic therapy.

Differential protein expression in white adipose tissue from obesity-prone and obesity-resistant mice in response to high fat diet and anti-obesity herbal medicines.

BACKGROUND: One of the most interesting issues in obesity research is why certain humans are obesity-prone (OP) while others are obesity-resistant (OR) upon exposure to a high-calorie diet. However, the pathways responsible for these phenotypic differences are still largely unknown. METHODS: In order to discover marker molecules determining susceptibility and/or resistance to obesity in response to high fat diet (HFD) or anti-obesity herbal medicine (TH), we conducted comparative proteomic analysis of white adipose tissue (WAT) from OP, OR, as well as TH-treated mice. RESULTS: OP mice fed HFD gained approximately 33% more body weight than OR mice, and TH significantly reduced body weight gain in HFD-fed mice by 30%. These mice were further subjected to proteomic analysis using two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Proteomic data revealed 59 spots that were differentially regulated from a total of 1,045 matched spots, and 57 spots of these were identified as altered WAT proteins between OP and OR mice by peptide mass finger printing. Interestingly, 45 proteins were similarly regulated in OR mice in response to TH treatment. Of these, 10 proteins have already been recognized in the context of obesity; however, other proteins involved in obesity susceptibility or resistance were identified for the first time in the present study. CONCLUSION: Our results suggest that TH actively contributed to body weight reduction in HFD-fed obese mice by altering protein regulation in WAT, and it was also found that TH-responsive proteins can be used as potent molecules for obesity treatment.

CDK4/6 inhibitors induce breast cancer senescence with enhanced anti-tumor immunogenic properties compared with DNA-damaging agents.

Since therapy-induced senescence (TIS) can either support or inhibit cancer progression, identifying which types of chemotherapeutic agents can produce the strongest anti-tumor TIS is an important issue. Here, cyclin-dependent kinase4/6 inhibitors (CDK4/6i)-induced senescence was compared to the TIS induced by conventional DNA-damaging agents. Despite both types of agents eliciting a similar degree of senescence, we observed increased expression of the senescence-associated secretory phenotype (SASP) and ligands related to pro-tumor immunity (IL6, CXCL8, TGFß, CD274, and CEACAM1) and angiogenesis (VEGFA) mainly in TIS induced by DNA-damaging agents rather than by CDK4/6i. Additionally, although all agents increased the expression of anti-tumor immunomodulatory proteins related to antigen presentation (MHC-I, B2M) and T cell chemokines (CXCL9, 10, 11), CDK4/6i-induced senescent cells still maintained this expression at a similar or even higher intensity than cells treated with DNA-damaging agents, despite the absence of nuclear factor-kappa-B (NF-κB) and p53 activation. These data suggest that in contrast with DNA-damaging agents, which augment the pro-tumorigenic microenvironment via pro-inflammatory SASP, CDK4/6i can generate TIS only with antitumor immunomodulatory proteins.

Cellular senescence is associated with the spatial evolution toward a higher metastatic phenotype in colorectal cancer.

In this study, we explore the dynamic process of colorectal cancer progression, emphasizing the evolution toward a more metastatic phenotype. The term "evolution" as used in this study specifically denotes the phenotypic transition toward a higher metastatic potency from well-formed glandular structures to collective invasion, ultimately resulting in the development of cancer cell buddings at the invasive front. Our findings highlight the spatial correlation of this evolution with tumor cell senescence, revealing distinct types of senescent tumor cells (types I and II) that play different roles in the overall cancer progression. Type I senescent tumor cells (p16INK4A+/CXCL12+/LAMC2-/MMP7-) are identified in the collective invasion region, whereas type II senescent tumor cells (p16INK4A+/CXCL12+/LAMC2+/MMP7+), representing the final evolved form, are prominently located in the partial-EMT region. Importantly, type II senescent tumor cells associate with local invasion and lymph node metastasis in colorectal cancer, potentially affecting patient prognosis.

C-terminus-deleted FoxM1 is expressed in cancer cell lines and induces chromosome instability.

Forkhead Box M1 (FoxM1) protein is a transcription factor and regulates cell cycle. It is commonly upregulated in human cancer tissue and correlated with poor prognosis, suggesting that the overexpression of FoxM1 plays a critical role in carcinogenesis. In this study, we report the identification and characterization of a new variant of FoxM1, which was first isolated from our laboratory in hepatoma cell lines. Compared with wild-type FoxM1, the new variant lacks of C-terminus of FoxM1 (FoxM1ΔC), which is a transactivation domain. Reverse transcription-polymerase chain reaction and western blot analysis demonstrated that FoxM1ΔC was highly expressed in a variety of cancer cell lines such as HepG2, HeLa, A549, MB231, EJ, U2OS, Hep3B and MCF7, but not expressed in normal human dermal fibroblast (HDF). Immunoprecipitation assay revealed that FoxM1ΔC interacted with wild-type FoxM1. Furthermore, FoxM1ΔC bound to FoxM1 targeted gene promoter region and correlated with dysregulation of wild-type FoxM1. FoxM1ΔC delayed G2/M to G1 progression of cell cycle, decreased Aurora B(T232) phosphorylation and increased chromosome centromere interspace. Finally, FoxM1ΔC induced instability of chromosome and formation of aneuploid cells within 1 month when expressed in HDF. In conclusion, FoxM1ΔC is expressed in cancer cells and dysregulates normal cell cycle and induces chromosome instability.

Benefit of systematic segmentectomy of the hepatocellular carcinoma: revisiting the dye injection method for various portal vein branches.

BACKGROUND: Systematic segmentectomy is useful in treating small hepatocellular carcinoma in the cirrhotic liver. However, accomplishment of an exact systematic segmentectomy still remains a challenging procedure because of the variable anatomy of portal branches. We evaluated the usefulness of the dye injection method for systematic segmentectomy, which focuses on the various patterns of portal vein (PV) branches feeding the tumor. METHODS: From January 2001 to May 2011, systematic segmentectomy by the dye injection method was performed in 70 patients. We evaluated the efficiency of systematic segmentectomy by ultrasonogram-guided dye injection into the portal branches that feed the tumor-bearing segments. The type of tumor-feeding PV branch, perioperative outcome, and survival rates were analyzed retrospectively. RESULTS: There were variations in the PV branches that fed the masses in 70 patients in whom the dye injection method for anatomical segmentectomy was tried. Forty masses (54.8%) were fed by a single main PV branch (type 1), 17 masses (23.3%) by a couple of PV branches (type 2), and 11 masses (15.1%) were supplied partially by single PV branch (type 3). In 5 patients (7.1%), masses were supplied by several small distributed PVs (type 4). For types 1 and 2, the tumor-bearing segments were resected anatomically with the help of staining; type 3 was partially stained and as the opposite side was not discrete, it was demarcated through counterstaining; and in type 4, dye injection could not be performed. Anatomical systematic segmentectomy was obtained in types 1 to 3; however, nonanatomical resection was inevitable for type 4. The 3- and 5-year overall survival rates were 80.5% and 67.2%, respectively, and the 3- and 5-year disease-free survival rates were 61.5% and 42.5%, respectively. The anatomical segmentectomy group showed better overall and disease-free survival than the nonanatomical group, even though it is not significant statistically. CONCLUSION: Systematic segmentectomy by the dye injection method overcomes the variation in PV tributaries in the segments and can be done according to the natural branching pattern of PVs.

Signals regulating necrosis of cardiomyoblast by BTG2(/TIS21/PC3) via activation of GSK3ß and opening of mitochondrial permeability transition pore in response to H2O2.

To investigate signal transduction pathway of cell death regulated by a tumor suppressor after oxidative stress, cardiomyoblasts were virally transfected with BTG2(/TIS21/PC3) (BTG2) and subsequently treated with H2O2. Heart muscle rarely expresses BTG2 unless oxidative stress occurs, however, ischemia induced BTG2 expression and necrosis, not apoptosis, of cardiomyoblasts. BTG2-expressioning cardiomyblasts showed impaired recoveries of survival kinases, Akt and Erk, thus sustaining GSK-3ß activity in 30 min of H2O2 exposure, in contrast to their rapid recoveries in LacZ control. The phenomenon was accompanied by the failure of ATP regeneration and the sustained activation of AMPK in the BTG2 expresser. Furthermore, H2O2 treatment markedly induced BTG2 translocation from nuclei to mitochondria along with cell death by cyclophilin D activation and mPTP opening. Exogenous and endogenous effect of BTG2 was confirmed by chemical inhibitors and BTG2-KO-MEF, respectively. Here, we suggest tumor suppressor, BTG2, as one of the regulators of necrosis in myocardium via inhibiting Akt/Erk, but activating GSK3ß and cyclophilin D, which resulted in mPTP opening in response to H2O2.