Wound Healing Potential of Low Temperature Plasma in Human Primary Epidermal Keratinocytes

BACKGROUND: Low temperature plasma (LTP) was recently shown to be potentially useful for biomedical applications such as bleeding cessation, cancer treatment, and wound healing, among others. Keratinocytes are a major cell type that migrates directionally into the wound bed, and their proliferation leads to complete wound closure during the cutaneous repair/regeneration process. However, the beneficial effects of LTP on human keratinocytes have not been well studied. Therefore, we investigated migration, growth factor production, and cytokine secretion in primary human keratinocytes after LTP treatment.METHODS: Primary cultured keratinocytes were obtained from human skin biopsies. Cell viability was measured with the EZ-Cytox cell viability assay, cell migration was evaluated by an in vitro wound healing assay, gene expression was analyzed by quantitative real-time polymerase chain reaction, and protein expression was measured by enzyme-linked immunosorbent assays and western blotting after LTP treatment.RESULTS: Cell migration, the secretion of several cytokines, and gene and protein levels of angiogenic growth factors increased in LTP-treated human keratinocytes without associated cell toxicity. LTP treatment also significantly induced the expression of hypoxia inducible factor-1α (HIF-1α), an upstream regulator of angiogenesis. Further, the inhibition of HIF-1α expression blocked the production of angiogenic growth factors induced by LTP in human keratinocytes.CONCLUSION: Our results suggest that LTP treatment is an effective approach to modulate wound healing-related molecules in epidermal keratinocytes and might promote angiogenesis, leading to improved wound healing.

Therapeutic potential of resveratrol for the treatment of type III Gaucher disease

Gaucher disease is the most common lysosomal storage disorder. Resveratrol is a natural polyphenol that possesses a wide range of beneficial effects, including anti-inflammatory, anti-oxidant, and neuroprotective activities. The aim of this study was to determine if resveratrol has a therapeutic effect on primary fibroblast cells derived from a patient with type III Gaucher disease. 3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were performed to determine the effect of resveratrol on cell survival. The expression levels of apoptosis-inducing factor (AIF), Bcl-2-associated X protein (Bax), caspase-3, acetyl-coenzyme A acetyltransferase 1 (ACAT1), E3- binding protein (E3BP), and citrate synthase (CS) were evaluated by western blotting to characterize the effect of resveratrol treatment on Gaucher disease cells. Thin-layer chromatography (TLC) was carried out to measure changes in glucosylceramide levels in resveratrol-treated patient cells. Resveratrol increased the viability of patient cells compared to that of untreated control cells. Resveratrol treatment dose-dependently decreased AIF, Bax, and cleaved caspase-3 levels, whereas ACAT1, E3BP, and CS expression dose-dependently increased. TLC analysis showed reduced levels of glucosylceramides in resveratrol-treated patient cells. These findings demonstrate that resveratrol can relieve cellular stress due to glucosylceramide accumulation, and suggest that it should be studied further as a new therapeutic approach for the treatment of Gaucher disease.

Channelopathies / 소아과

Channelopathies are a heterogeneous group of disorders resulting from the dysfunction of ion channels located in the membranes of all cells and many cellular organelles. These include diseases of the nervous system (e.g., generalized epilepsy with febrile seizures plus, familial hemiplegic migraine, episodic ataxia, and hyperkalemic and hypokalemic periodic paralysis), the cardiovascular system (e.g., long QT syndrome, short QT syndrome, Brugada syndrome, and catecholaminergic polymorphic ventricular tachycardia), the respiratory system (e.g., cystic fibrosis), the endocrine system (e.g., neonatal diabetes mellitus, familial hyperinsulinemic hypoglycemia, thyrotoxic hypokalemic periodic paralysis, and familial hyperaldosteronism), the urinary system (e.g., Bartter syndrome, nephrogenic diabetes insipidus, autosomal-dominant polycystic kidney disease, and hypomagnesemia with secondary hypocalcemia), and the immune system (e.g., myasthenia gravis, neuromyelitis optica, Isaac syndrome, and anti-NMDA [N-methyl-D-aspartate] receptor encephalitis). The field of channelopathies is expanding rapidly, as is the utility of molecular-genetic and electrophysiological studies. This review provides a brief overview and update of channelopathies, with a focus on recent advances in the pathophysiological mechanisms that may help clinicians better understand, diagnose, and develop treatments for these diseases.

The large-conductance calcium-activated potassium channel holds the key to the conundrum of familial hypokalemic periodic paralysis / 소아과

PURPOSE: Familial hypokalemic periodic paralysis (HOKPP) is an autosomal dominant channelopathy characterized by episodic attacks of muscle weakness and hypokalemia. Mutations in the calcium channel gene, CACNA1S, or the sodium channel gene, SCN4A, have been found to be responsible for HOKPP; however, the mechanism that causes hypokalemia remains to be determined. The aim of this study was to improve the understanding of this mechanism by investigating the expression of calcium-activated potassium (KCa) channel genes in HOKPP patients. METHODS: We measured the intracellular calcium concentration with fura-2-acetoxymethyl ester in skeletal muscle cells of HOKPP patients and healthy individuals. We examined the mRNA and protein expression of KCa channel genes (KCNMA1, KCNN1, KCNN2, KCNN3, and KCNN4) in both cell types. RESULTS: Patient cells exhibited higher cytosolic calcium levels than normal cells. Quantitative reverse transcription polymerase chain reaction analysis showed that the mRNA levels of the KCa channel genes did not significantly differ between patient and normal cells. However, western blot analysis showed that protein levels of the KCNMA1 gene, which encodes KCa1.1 channels (also called big potassium channels), were significantly lower in the membrane fraction and higher in the cytosolic fraction of patient cells than normal cells. When patient cells were exposed to 50 mM potassium buffer, which was used to induce depolarization, the altered subcellular distribution of BK channels remained unchanged. CONCLUSION: These findings suggest a novel mechanism for the development of hypokalemia and paralysis in HOKPP and demonstrate a connection between disease-associated mutations in calcium/sodium channels and pathogenic changes in nonmutant potassium channels.

Altered expression of potassium channel genes in familial hypokalemic periodic paralysis

We analyzed the mRNA expression patterns of major potassium channel genes to determine the mechanism of hypokalemia in familial hypokalemic periodic paralysis. We used quantitative RT-PCR to examine the mRNA levels of both inward (KCNJ2, KCNJ6, and KCNJ14) and delayed rectifi er (KCNQ1 and KCNA2) potassium channel genes in skeletal muscle cells from both normal and patient groups, prior to and after exposure to 4 mM and 50 mM potassium buffers. Quantitative RT-PCR analysis revealed no changes in the mRNA levels of these genes in normal and patient cells on exposure to 4 mM potassium buffer. However, after exposure to 50 mM potassium buffer, which was used to induce depolarization, normal cells showed a signifi cant decrease in KCNJ2, KCNJ6, and KCNJ14 expression, but no change in KCNQ1 and KCNA2 expression. In contrast, patient cells showed no change in KCNJ2 and KCNJ6 expression, but an increase in KCNJ14 expression. Furthermore, KCNQ1 and KCNA2 showed decreased expression. We found that the expression levels of both inward and delayed rectifi er potassium channel genes in patient cells differ from those in normal cells. Altered potassium channel gene expression in patient cells may suggest a possible mechanism for hypokalemia in familial hypokalemic periodic paralysis.

Hyperkalemic periodic paralysis and paramyotonia congenita caused by a de novo mutation in the SCN4A gene

Familial hyperkalemic periodic paralysis is an autosomal-dominant channelopathy characterized by reversible paralysis associated with episodic hyperkalemia. Mutations in the skeletal muscle voltage-gated sodium channel gene (SCN4A) have been reported to be responsible for this disorder. Paramyotonia congenita is also caused by mutations in the SCN4A gene. Here, we report the case of a 17-year-old boy who presented with both hyperkalemic periodic paralysis and paramyotonia congenita. A molecular analysis of the SCN4A gene revealed a heterozygous T>C transition at nucleotide 2078, leading to an Ile693Thr mutation. This mutation was absent in the patient’s parents supporting a de novo Ile693Thr mutation in our patient.

Transient neonatal diabetes mellitus caused by a de novo ABCC8 gene mutation / 소아과

Transient neonatal diabetes mellitus (TNDM) is a rare form of diabetes mellitus that presents within the first 6 months of life with remission in infancy or early childhood. TNDM is mainly caused by anomalies in the imprinted region on chromosome 6q24; however, recently, mutations in the ABCC8 gene, which encodes sulfonylurea receptor 1 (SUR1), have also been implicated in TNDM. Herein, we present the case of a male child with TNDM whose mutational analysis revealed a heterozygous c.3547C>T substitution in the ABCC8 gene, leading to an Arg1183Trp mutation in the SUR1 protein. The parents were clinically unaffected and did not show a mutation in the ABCC8 gene. This is the first case of a de novo ABCC8 gene mutation in a Korean patient with TNDM. The patient was initially treated with insulin and successfully switched to sulfonylurea therapy at 14 months of age. Remission of diabetes had occurred at the age of 16 months. Currently, the patient is 21 months old and is euglycemic without any insulin or oral hypoglycemic agents. His growth and physical development are normal, and there are no delays in achieving neurological and developmental milestones.

Hematologic Profiles and Eosinophilia in Very Low Birth Weight Infants / 대한주산의학회잡지

PURPOSE: This study investigated the pattern of hematologic profile and eosinophilia for a month after birth in very low birth weight (VLBW) infants. METHODS: The medical records of 141 VLBW infants (birth weight, or =700/mm3). RESULTS: Overall, 50.4% of all infants had at least one instance of eosinophilia for a month after birth. There were 50.7% with moderate eosinophilia (1,000-2,999/mm3). White blood cell (WBC) counts and absolute neutrophil count (ANC) climaxed on 7th day of life, whereas eosinophilia mainly occurred on 21st day of life. The demographic data and perinatal characteristics of infants with and without eosinophilia were compared. Prevalence of eosinophilia was associated with gestational age and total parenteral nutrition on 21st day of life; total parenteral nutrition and transfusion on 28th day of life. Eosinophilia was closely associated with transfusion on logistic regression analysis (P<0.05). CONCLUSION: Eosinophilia in VLBW infants occurs mainly on 21st day of life. Eosinophil counts showed a separate trend different from WBC counts and ANC. Transfusion was significantly associated with eosinophilia.

Familial hyperkalemic periodic paralysis caused by a de novo mutation in the sodium channel gene SCN4A / 소아과

Familial hyperkalemic periodic paralysis (HYPP) is an autosomaldominant channelopathy characterized by transient and recurrent episodes of paralysis with concomitant hyperkalemia. Mutations in the skeletal muscle voltage-gated sodium channel gene SCN4A have been reported to be responsible for this disease. Here, we report the case of a 16-year-old girl with HYPP whose mutational analysis revealed a heterozygous c.2111C>T substitution in the SCN4A gene leading to a Thr704Met mutation in the protein sequence. The parents were clinically unaffected and did not have a mutation in the SCN4A gene. A de novo SCN4A mutation for familial HYPP has not previously been reported. The patient did not respond to acetazolamide, but showed a marked improvement in paralytic symptoms upon treatment with hydrochlorothiazide. The findings in this case indicate that a de novo mutation needs to be considered when an isolated family member is found to have a HYPP phenotype.

Familial hyperkalemic periodic paralysis caused by a de novo mutation in the sodium channel gene SCN4A / 소아과

Familial hyperkalemic periodic paralysis (HYPP) is an autosomaldominant channelopathy characterized by transient and recurrent episodes of paralysis with concomitant hyperkalemia. Mutations in the skeletal muscle voltage-gated sodium channel gene SCN4A have been reported to be responsible for this disease. Here, we report the case of a 16-year-old girl with HYPP whose mutational analysis revealed a heterozygous c.2111C>T substitution in the SCN4A gene leading to a Thr704Met mutation in the protein sequence. The parents were clinically unaffected and did not have a mutation in the SCN4A gene. A de novo SCN4A mutation for familial HYPP has not previously been reported. The patient did not respond to acetazolamide, but showed a marked improvement in paralytic symptoms upon treatment with hydrochlorothiazide. The findings in this case indicate that a de novo mutation needs to be considered when an isolated family member is found to have a HYPP phenotype.

A Case of Neonatal Lupus Erythematosus and Retrospectively Diagnosed with Asymtomatic Maternal Lupus Erythematosus

Neonatal lupus erythematosus (NLE) is a rare disease characterized by typical clinical features and the transplacental passage of maternal autoantibodies, particularly anti-SSA/Ro. The major clinical manifestations are cutaneous lupus lesions, congenital heart block, hematological disorders, and hepatobiliary diseases. We report a case of NLE presenting with multiple round and oval target-like erythematous skin lesions and abnormal liver function, born to a clinically asymptomatic mother whose diagnosis was made retrospectively only after her newborn's diagnosis. Both the infant and the mother were positive for the anti-SSA/Ro and anti-SSB/La antibodies.

A Case of Penile Cutaneous Metastasis from Gastric Carcinoma / 대한피부과학회지

Metastatic carcinoma of the penis is rare and in most cases is associated with primary malignancy of the pelvic organs with widespread metastasis. A 64-year-old man presented with a 2 month history of skin-colored papules on his penis. He also had a history of total gastrectomy and chemotherapy due to advanced gastric carcinoma. Skin biopsy from the lesion revealed atypical polygonal cells with hyperchromatic & pleomorphic nuclei in the dermis. Results of immunohistochemical staining showed a positive reaction of CK7, and a negative reaction of CK20, PSA, and CD68. We report herein a case of metastatic cancer of the penis from gastric carcinoma.

Immunohistochemical Study on the Expression of Desmoglein-3 in Fetal Skin Development / 대한피부과학회지

BACKGROUND: Desmogleins are calcium-dependent transmembrane glycoproteins of the desmosome that form an import component of the junction complexes of epithelial cells. Desmogleins are involved in maintaining the structural integrity of tissues. So far, four different desmogleins (Dsg1, Dsg2, Dsg3 and Dsg4) have been identified. OBJECTIVE: The purpose of this study was to observe the distribution pattern of desmoglein-3 in the fetal skin during development. METHODS: Skin was obtained from the sole, scalp and lip of 34 fetuses that ranged in age from 10 to 39 weeks of gestational age. Immunohistochemical staining was performed on the paraffin embedded tissue using anti-human monoclonal antibody against the desmoglein-3. RESULTS: The expression of desmoglein-3 in the epidermis appeared in the basal layer of the sole at the 10th week of gestation age. Thereafter, a stronger expression was noted in the middle layer of the sole and scalp epidermis. The basal layer had a stronger expression than did the other layers of the epidermis, followed by the middle and superficial layers. A stronger expression of desmoglein-3 in hair was noted in the outer root sheath, the bulge cells and the eccrine duct cells. The expression of desmoglein-3 in the lip mucosa was strong in both the basal and middle layers, while the skin side of the mucosa showed a stronger expression in basal layer. CONCLUSION: These results suggested that desmoglein-3 plays an important role in the development and differentiation of the epidermis and skin adnexa in the fetal stage, and especially in basal and suprabasal layers.

An atypical phenotype of hypokalemic periodic paralysis caused by a mutation in the sodium channel gene SCN4A / 소아과

Familial hypokalemic periodic paralysis is an autosomal-dominant channelopathy characterized by episodic muscle weakness with hypokalemia. The respiratory and cardiac muscles typically remain unaffected, but we report an atypical case of a family with hypokalemic periodic paralysis in which the affected members presented with frequent respiratory insufficiency during severe attacks. Molecular analysis revealed a heterozygous c.664 C>T transition in the sodium channel gene SCN4A, leading to an Arg222Trp mutation in the channel protein. The patients described here presented unusual clinical characteristics that included a severe respiratory phenotype, an incomplete penetrance in female carriers, and a different response to medications.

Phosphatidylcholine Formula for the Treatment of Lipoma / 대한피부과학회지

BACKGROUND: Lipomas are benign neoplasms composed of mature fat cells. Current treatments are invasive and carry the risk of scarring. For this reason, phosphatidylcholine (PPC) formula has been widely used to treat areas of localized fat accumulation. However, there are few reports on injection lipolysis with injection of PPC formula. OBJECTIVE: The purpose of this study was to investigate whether injection lipolysis with PPC formula is an effective therapeutic option for patients with lipoma. METHODS: Twenty-two lipomas in 7 patients were included. Three sessions at intervals of 6 to 8 weeks were done. Sonographic measurements of lipoma size were repeated before each session. Any side-effects and the patients' satisfaction were noted. In one lipoma, histological changes after lipolysis were described. RESULTS: After three sessions, a significant reduction in lipoma size of 47.4% was achieved. Histology showed a fibrosis with loss of adipocytes. Inflammation occurred in one case, so the patient was excluded from this study. Transient burning sensation and swelling at the injection site were the most common adverse effects. No severe side-effects or systemic reactions were observed. CONCLUSION: Treatment of lipoma with PPC formula has the potential to be a patient-friendly and well-tolerated therapy that can achieve cosmetically pleasing treatment outcomes.

Langerhans Cell Histiocytosis That Improved with Oral Isotretinoin Therapy / 대한피부과학회지

Langerhans cell histiocytosis (LCH) is a reactive disease in which abnormal Langerhans cells accumulate in various body sites. We report here on a 51-year-old male patient with LCH that was classified as single-system disease (restricted LCH). The skin lesions were a small, deep ulcer in the right inguinal area and multiple erythematous papules scattered on the lower abdomen, and the patient had these lesions for 1 year. The histopathologic examination revealed a dense histiocytic infiltration in the dermis, and most of the cells showed the characteristics of "LCH" cells. The immunohistochemistry for S-100 protein and CD1a complex all showed positive results. The patient was much improved after surgical excision of the inguinal ulcer and administering oral isotretinoin (20 mg daily) for 8 months, and there was no recurrence. We think retinoid is an effective treatment option for LCH, especially for single system disease.

A Case of Primary Cutaneous Mucinous Carcinoma with Neuroendocrine Differentiation

Primary cutaneous mucinous carcinoma is a rare malignant tumor that originates from the deepest portion of the eccrine sweat duct. Common sites of involvement are the face and scalp. Biopsy shows dermal epithelial cell islands embedded in mucin pools separated by fibrous septae. It is difficult to differentiate this tumor histologically from metastatic adenocarcinoma. Recurrence after excision is common but metastases are rare. We report a primary cutaneous mucinous carcinoma with neuroendocrine differentiation on the right cheek of a 63-year-old man.

Multiple, Giant Soft Fibromas in the Labium Majus / 대한피부과학회지

Soft fibromas are benign fibrous tissue tumors of the dermis. In general, they appear as small, soft, pedunculated or sessile tumors and they are mostly located on the axillae, lateral neck and inguinal region. Although large sized soft fibromas may occur in the groin or upper thighs, and especially in patients with diabetes, they rarely exceed 2 cm in size. We report herein a case of multiple giant soft fibromas, including an unusually huge, giant lesion in the labium majus of a healthy, middle-aged woman, and such fibromas on the labium majus are very rare.

A Case of Malignant Fibrous Histiocytoma That Metastasized to the Skin / 대한피부과학회지

Malignant fibrous histiocytoma (MFH) is the most common soft tissue sarcoma. It is regarded as an undifferentiated pleomorphic sarcoma with an unproven histogenesis. It most commonly occurs in the skeletal muscles of the extremities or the retroperitoneum of adults. It often metastasizes to the lungs, lymph nodes, bones and liver, but rarely to the skin. We report here a case of malignant fibrous histiocytoma that metastasized to the skin.

A Case of Syringoma Confined to Both Forearms and the Dorsa of the Hands / 대한피부과학회지

Syringomas confined to the distal extremities are very rare. A 62-year-old man presented with an 8-month history of multiple, light brown, flat-topped papules on both forearms and the dorsa of the hands. Histopathologic examination showed multiple dilated eccrine ducts lined by two rows of epithelial cells. There has currently been less than 10 reported cases of syringoma confined to both forearms and the dorsa of the hands in the world medical literature.