Extraregional lymph node recurrence of stage IA1 squamous cell carcinoma of the uterine cervix after initial surgery: two case studies.

Objective: Lymph node recurrence is extremely rare in cases of stage IA1 squamous cell carcinoma (SCC) of the uterine cervix without lymphovascular space invasion (LVSI). We present two cases of extraregional lymph node recurrence after initial surgery for stage IA1 SCC of the uterine cervix without LVSI. Patients: Both patients initially underwent hysterectomy and developed recurrent extraregional lymph nodes within a few years postoperatively. Case 1: The patient showed no symptoms of recurrence, and follow-up computed tomography (CT) for evaluation of gallstones revealed a para-aortic lymph node (9 mm). The patient subsequently underwent serum SCC antigen testing and CT and was diagnosed with recurrence. Case 2: The patient noticed a right inguinal node swelling, which was evaluated using CT. Both patients survived without relapse for 8 and 4 years, respectively. Conclusion: Although stage IA1 SCC of the uterine cervix without LVSI is associated with a low risk of lymph node recurrence, oncologists should consider the possibility of recurrence in such cases. Evaluation for recurrence is difficult in asymptomatic patients. Serum SCC antigen testing may be a useful biochemical marker before imaging for early detection of recurrence, even in asymptomatic patients.

Relevance of Solidification Kinetics for Enhanced Thermoelectric Performance in Al-Doped Higher Manganese Silicides.

The solidification kinetics of an alloy from its liquid state forms an underlying basis for microstructural engineering, wherein the state of thermodynamic equilibrium associated with the melt-grown crystal and the quenched amorphous solid denotes the two limits for crystallinity in the alloy synthesis. In this study, we report the implication of the crystalline state on the thermal and electrical transport properties of partially substituted Mn(Si1-xAlx)γ by comparing the single crystals melt-grown by the Bridgman method, and polycrystals synthesized from melt spinning (MS) and subsequent spark plasma sintering (SPS). The rapidly solidified alloys exhibited nanocrystalline microstructures in MS ribbons, while melt-grown single crystals displayed characteristics evolution of MnSi striations with limited solubility of Al. It was observed that Al as a p-type dopant enhances the carrier concentration and electrical conductivity, while nanocrystallinity in MS + SPS polycrystals and secondary phases in monocrystals were effective in enhancing the phonon scattering. Maximum zT values of ∼0.54 (±0.05) at 823 K and 0.75 (±0.05) at 873 K were attained for the single crystal (directed perpendicular to the c-axis) and melt-spun polycrystals (along the in-plane direction), respectively. These results present the efficacy of aliovalent Al substitution and demonstrate the critical role of the solidification kinetics in optimizing the carrier concentration and enhancing the phonon scattering in higher manganese silicide crystals for thermoelectric applications.

Case of heterotopic cervical pregnancy and total placenta accreta after artificial cycle frozen-thawed embryo transfer.

Case: A 39-year-old woman presented with a genital hemorrhage at 5 weeks of gestation after an artificial cycle double frozen-thawed embryo transfer. She was diagnosed with a cervical heterotopic pregnancy. Although hormone supplementation was discontinued to terminate the pregnancy at 5 weeks of gestation, the intrauterine and cervical gestational sacs continued to develop. Outcome: The cervical gestational sac was surgically removed and the intrauterine pregnancy continued uneventfully, except for vasa previa. At 36 weeks of gestation, the patient underwent a cesarean section and gave birth to a healthy female infant. At the delivery, massive bleeding occurred and a hysterectomy was performed due to total placenta accreta. Conclusion: This case provides a novel example of a near-term delivery after a cervical heterotopic pregnancy and emphasizes the need for intensive care, even after the successful management of a cervical pregnancy. Most importantly, the present case implies a possible link between hormonal withdrawal and abnormal placentation.

Three cases of reconstruction with a simply designed bilobed flap after excision of squamous cell carcinomas of the lateral eyebrow areas.

Squamous cell carcinoma is frequently observed on the lateral eyebrow, one of the most difficult sites for one-stage reconstruction because of its location close to the eyelids. The reconstruction of defects by the single-stage flap is advantageous for elderly patients because of its good functional and aesthetic outcome. We surgically treated three cases of squamous cell carcinoma on this area and performed reconstruction with a simply designed bilobed flap, all of which resulted in favorable outcomes. We therefore report on the usefulness of this reconstructive method for the lateral eyebrow defects.

A Study on the Alterations in Skin Viscoelasticity before and after an Intradermal Administration of Growth Factor.

BACKGROUND: While photo-aging is believed to be preventable by the complete blockage of ultraviolet rays, there is no epoch-making method except sing fillers or autologous fat injection, for rejuvenating the skin once it has aged. OBJECTIVE: Our group developed a new method for rejuvenating aged skin by the direct intradermal injection of basic fibroblast growth factor, the first method of its kind in the world. In this paper we report the results of long-term follow-up observations and alterations in skin viscoelasticity before and after this treatment. MATERIALS AND METHODS: A single dose of growth factor was injected directly into aged skin of the dorsal surface of the hand intradermally. The skin viscoelasticity of 50 treated cases was measured by a cutometer just before the treatment and at 1, 3, 6, and 9 months after treatment, respectively. RESULTS: WE OBSERVED THE FOLLOWING REJUVENATING EFFECTS: improved skin softness, gradual improvement of turgor, improved thickness of atrophied skin, and greatly improved viscoelasticity which reveals the improvement of biomechanical properties of the treated aged skin. According to the comparisons of viscoelasticity between pre- and post-treatment, the rejuvenated changes of R2 and R7 values were comparable to an age difference of more than 20 years. CONCLUSION: This method was confirmed to have excellent effects in rejuvenating aged skin safely and reliably including biomechanical properties. With this advance, we expect conventional non-physiological skin rejuvenating treatments to be replaced by a much more fundamental method using one-time injections of the growth factor.

Melanoma-targeted chemo-thermo-immuno (CTI)-therapy using N-propionyl-4-S-cysteaminylphenol-magnetite nanoparticles elicits CTL response via heat shock protein-peptide complex release.

Melanogenesis substrate, N-propionyl-4-S-cysteaminylphenol (NPrCAP) is specifically taken up by melanoma cells and inhibits their growth by producing cytotxic free radicals. By taking advantage of this unique chemical agent, we have established melanoma-targeting intracellular hyperthermia by conjugating NPrCAP with magnetite nanoparticles (NPrCAP/M) upon exposure to an alternating magnetic field (AMF). This treatment causes cytotoxic reaction as well as heat shock responses, leading to elicitation of antitumor immune response, which was proved by tumor rechallenge test and CTL induction. We found the level of heat shock protein 72 (Hsp72) to be increased in the cell lysate and culture supernatant after intracellular hyperthermia. Melanoma-specific CD8(+) T-cell response to dendritic cells loaded with hyperthermia-treated tumor lysate was enhanced when compared with non-treated tumor lysate. When heat shock protein, particularly Hsp72, was immuno-depleted from hyperthermia-treated tumor cell lysate, specific CD8(+) T-cell response was abolished. Thus, it is suggested that antitumor immune response induced by hyperthermia using NPrCAP/M is derived from the release of HSP-peptide complex from degraded tumor cells. Therefore, this chemo-thermo-immuno (CTI)-therapy might be effective not only for primary melanoma but also for distant metastasis because of induction of systemic antimelanoma immune responses.

The mechanisms underlying fibroblast apoptosis regulated by growth factors during wound healing.

While investigating the mechanisms underlying cell death during wound healing processes, we uncovered the pro-apoptotic effects of basic fibroblast growth factor (bFGF) on granulation tissue fibroblasts following pretreatment with transforming growth factor (TGF)-beta1 in vitro. bFGF induced caspase-3 activation and apoptosis in TGF-beta1-pretreated granulation tissue-derived fibroblasts (GF-1) following bFGF treatment for 48 and 96 h. In contrast, fibroblasts that had been treated in the same manner and that originated from the uninjured dermis did not display apoptosis, indicating that the mechanisms underlying apoptosis events in fibroblasts that originate from normal dermal and wound tissues differ. In this process, we also found that bFGF inhibited Akt phosphorylation at serine 473 and induced a rapid loss of phosphorylation of focal adhesion kinase (FAK) at tyrosine 397 in pretreated GF-1 cells, an event that coincided with the dissociation of phosphorylated FAK from the focal adhesions. Therefore, inhibition of survival signals relayed via the disrupted focal adhesion structures and inactivated Akt following bFGF treatment may lead to apoptosis in GF-1 cells pretreated with TGF-beta1. Pretreatment of GF-1 with TGF-beta1 followed by the addition of bFGF resulted in significantly greater inhibition of phosphorylation of Akt and FAK compared to treatment with TGF-beta1 or bFGF alone. The combinatorial treatment also led to proteolysis of FAK and inhibition of FAK and Akt protein expression in GF-1 cells. These findings demonstrated a significant role for the two cytokines in apoptosis of granulation tissue fibroblasts during wound healing. In vivo studies also confirmed a marked decline in phosphorylation and protein expression of Akt and FAK in bFGF-injected skin wounds. These results led to the hypothesis that temporal activation of TGF-beta1 and bFGF at the injury site promotes apoptosis in granulation tissue fibroblasts, an event that is critical for the termination of proliferative granulation tissue formation.

Growth inhibition of re-challenge B16 melanoma transplant by conjugates of melanogenesis substrate and magnetite nanoparticles as the basis for developing melanoma-targeted chemo-thermo-immunotherapy.

Melanogenesis substrate, N-propionyl-cysteaminylphenol (NPrCAP), is selectively incorporated into melanoma cells and inhibits their growth by producing cytotoxic free radicals. Magnetite nanoparticles also disintegrate cancer cells and generate heat shock protein (HSP) upon exposure to an alternating magnetic field (AMF). This study tested if a chemo-thermo-immunotherapy (CTI therapy) strategy can be developed for better management of melanoma by conjugating NPrCAP on the surface of magnetite nanoparticles (NPrCAP/M). We examined the feasibility of this approach in B16 mouse melanoma and evaluated the impact of exposure temperature, frequency, and interval on the inhibition of re-challenged melanoma growth. The therapeutic protocol against the primary transplanted tumor with or without AMF exposure once a day every other day for a total of three treatments not only inhibited the growth of the primary transplant but also prevented the growth of the secondary, re-challenge transplant. The heat-generated therapeutic effect was more significant at a temperature of 43 degrees C than either 41 degrees C or 46 degrees C. NPrCAP/M with AMF exposure, instead of control magnetite alone or without AMF exposure, resulted in the most significant growth inhibition of the re-challenge tumor and increased the life span of the mice. HSP70 production was greatest at 43 degrees C compared to that with 41 degrees C or 46 degrees C. CD8(+)T cells were infiltrated at the site of the re-challenge melanoma transplant.

De novo follicular regeneration of the skin by wingless int 3 and bone morphogenetic protein 2 genes introduced into dermal fibroblasts and fibroblast growth factor-2 protein.

In this study, we regenerated skin and its appendages by transplanting cultured normal dermal fibroblasts, into which morphogen genes had been introduced. We cultured normal dermal fibroblasts obtained from Fisher 344 rats on the surface of hydroxyapatite beads, and then adsorbed them onto the surface of a collagen sponge, which was transplanted into a full-thickness skin defect prepared on the backs of rats. Before transplantation, genes were introduced into the dermal fibroblasts via adenovirus vector (ad)-bone morphogenetic protein 2 and ad-wingless int 3 genes in addition to fibroblast growth factor-2 protein. By Week 4, the appearance of follicle germs or primitive hair germs was observed only in the ad-bone morphogenetic protein 2+ad-wingless int 3 combined with the fibroblast growth factor-2 protein group. By Week 16, in that same group, hair follicles having mature pilosebaceous systems with equally spaced localization had formed in the ulcer wound.

N-propionyl-cysteaminylphenol-magnetite conjugate (NPrCAP/M) is a nanoparticle for the targeted growth suppression of melanoma cells.

A magnetite nanoparticle, NPrCAP/M, was produced for intracellular hyperthermia treatment of melanoma by conjugating N-propionyl-cysteaminylphenol (NPrCAP) with magnetite and used for the study of selective targeting and degradation of melanoma cells. NPrCAP/M, like NPrCAP, was integrated as a substrate in the oxidative reaction by mushroom tyrosinase. Melanoma, but not non-melanoma, cells incorporated larger amounts of iron than magnetite from NPrCAP/M. When mice bearing a B16F1 melanoma and a lymphoma on opposite flanks were given NPrCAP/M, iron was observed only in B16F1 melanoma cells and iron particles (NPrCAP/M) were identified within late-stage melanosomes by electron microscopy. When cells were treated with NPrCAP/M or magnetite and heated to 43 degrees C by an external alternating magnetic field (AMF), melanoma cells were degraded 1.7- to 5.4-fold more significantly by NPrCAP/M than by magnetite. Growth of transplanted B16 melanoma was suppressed effectively by NPrCAP/M-mediated hyperthermia, suggesting a clinical application of NPrCAP/M to lesional therapy for melanoma. Finally, melanoma cells treated with NPrCAP/M plus AMF showed little sub-G1 fraction and no caspase 3 activation, suggesting that the NPrCAP/M-mediated hyperthermia induced non-apoptotic cell death. These results suggest that NPrCAP/M may be useful in targeted therapy for melanoma by inducing non-apoptotic cell death after appropriate heating by the AMF.

Basic fibroblast growth factor reduces scar formation in acute incisional wounds.

In order to identify a means to reduce scar formation of the skin after incision, this study examined the effect of local administration of basic fibroblast growth factor (bFGF) in humans. bFGF was administered to a sutured wound immediately after an operation. The drug was injected once into the dermis of the margins of wounds using a 27G needle or rinsing after performing dermostitches. The lengths of the treated wounds varied from 1 to 32 cm, and the subjects were 2-86 years old. Sutured wounds after excision of skin tumors from the face, trunk, and limbs and sutured wounds such as those at the donor sites of full-thickness skin grafts were treated with low-dose bFGF injections (0.1 microg/cm wound; Group 2), high-dose bFGF injections (1 microg/cm wound; Group 3), and rinsed with high-dose bFGF (1 microg/cm wound; Group 4). No patient treated with bFGF had hypertrophic scars, while some patients had hypertrophic or very wide scars in the control group (Group 1), and the ratios of minimum scarring of Group 2 (p<0.001), Group 3 (p<0.0001), and Group 4 (p<0.0001) were statistically significantly higher than those of Group 1. Postoperative administration of bFGF inhibited hypertrophic scarring and widening of remaining scars without any serious side effects.

Basic fibroblast growth factor in an artificial dermis promotes apoptosis and inhibits expression of alpha-smooth muscle actin, leading to reduction of wound contraction.

To clarify the mechanisms underlying declines in wound contraction caused by basic fibroblast growth factor (bFGF) and the role of autologous fibroblasts in modulating wound healing, we have examined the expression of alpha-smooth muscle actin (alpha-SMA) and apoptosis in a model of wound healing using collagen sponges with and without bFGF (1 microg) and/or fibroblasts (1 x 10(6) cells/cm(2)) applied to experimentally produced full-thickness skin wounds in rats (n=10 for each group). At 7 days postoperatively, wounds filled with a fibroblast-seeded collagen sponge (fibroblast-seeded group) displayed a greater area of collagen sponge and a smaller area of fibroblasts compared with control wounds filled with collagen sponge alone (control group). Therefore, seeding of fibroblasts in the dermal substitute might retard degradation of the collagen sponge, inhibiting fibroblast infiltration into the substitute. By day 14, wounds filled with bFGF-treated collagen sponge without fibroblast seeding (bFGF group) displayed decreased alpha-SMA expression and significantly increased apoptosis compared with other wounds. Double staining revealed that apoptosis in alpha-SMA-positive fibroblastic cells was significantly increased in the bFGF group, suggesting that bFGF treatment is a potent stimulator of myofibroblast apoptosis. Furthermore, morphometric analysis demonstrated the significant decrease in the level of wound contraction and the degree of mature collagen bundle formation in the bFGF group by day 42. The bFGF group also showed increased bFGF expression in macrophages by day 28. These results suggest that bFGF administration to an artificial dermis promotes apoptosis of alpha-SMA-positive fibroblastic cells and inhibits alpha-SMA expression in the treated wound, thus reducing wound contraction.

Papulonecrotic tuberculid-like eruptions after Bacille Calmette-Guerin vaccination.

One of the specific skin lesions occurring after Bacille Calmette-Guerin (BCG) vaccination is generalized tuberculid-like eruptions, which occur rarely, but have a tendency to heal spontaneously. Their pathogenesis and relationship to "true" tuberculids are poorly understood. This report presents a case of a 6-month-old girl who developed generalized papulonecrotic tuberculid-like eruptions after BCG vaccination. The skin lesions healed spontaneously in 3 months. Culture of blood, gastric juice and reverse transcription polymerase chain reaction (RT-PCR) of papulonecrotic skin biopsies were all negative for Mycobacterium tuberculosis. Histopathology of papulonecrotic eruptions revealed marked epidermal necrosis, perivascular lymphocytic infiltrates and epidermotropic infiltration of lymphocytes showing markers of CD3(+) lymphocytes (90-95% of all infiltrating cells), CD4(+) (40-50%), CD8(+) (40-50%), and CD45RO(+) (70%). In contrast, the BCG vaccination site revealed intradermal granuloma with epithelioid cells, occasional giant cells and infiltration of lymphocytes consisting of CD3(+) (60-70%), CD4(+) (40-50%), CD8(+) (30-40%), CD45RO(+) (40%), CD79a(+) (30-40%), and CD20(+) (20-30%). Our patient did not reveal any signs indicative of tuberculosis. Papulonecrotic lesions were therefore called papulonecrotic tuberculid-like eruptions, rather than tuberculids, that occurred after BCG vaccination and appeared to derive from a hypersensitive reaction mediated by immune lymphocytic infiltration.

The real-time, three-dimensional analyses of benign and malignant skin tumors by confocal laser scanning microscopy.

BACKGROUND: In obtain images of skin tumors non-invasively with real-time, confocal laser scanning microscope (CLSM) is introduced. OBJECTIVE: Reconstructed images of given horizontal sections were converted into three-dimensions using the data set of a large number of tomograms in the horizontal directions. METHODS: To develop the multiplaner reconstruction images of skin tumors in vertical directions and three-dimensionally reconstructed images of tumors will be obtained from the continuously collected horizontal image data sets. RESULTS: Three-dimensional analyses of the skin tumors from reconstructed images of the CLSM scanning have provided the information as to their physiological characteristics as well as the extent of deep invasion in real-time with non-invasive manner. High performance three-dimensional conversion software was effective in displaying three-dimensional construction of skin tumors. CONCLUSION: The CLSM scanning images followed by three-dimensional reconstruction using them can provide the real-time and non-invasive diagnoses of skin tumors and analyze the radial growth phase of tumors and the three-dimensional growth characteristics.

Reconstruction method with a newly-designed bilobed flap after excision of tumours of the skin.

We have devised a bilobed skin flap for reconstruction after excision of small skin tumours. The sutured part serves as a zig-zag that leads to only slight postoperative contracture of the scar. The rotation centre of the flap is nearer to the affected area than other conventional bilobed flaps, resulting in less dog-ear deformity and distortion of tissue.

Activated caspase expression and apoptosis increase in keloids: cytochrome c release and caspase-9 activation during the apoptosis of keloid fibroblast lines.

To characterize apoptosis in keloids and the mechanisms responsible for this process, the expression of activated caspase-9 and -3 in fibroblasts obtained from keloids was analyzed. Immunohistochemistry revealed that the number of fibroblasts positive for terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) or activated caspase-9 or -3 was low but was significantly higher in keloid tissues than in normal scar tissues. Significant relationships between the number of caspase-positive fibroblasts and TUNEL-positive fibroblasts suggested that the activation of caspase-9 and -3 induces apoptosis in a subpopulation of keloid fibroblasts. All keloid fibroblast cell lines established in this study showed activation of caspase-9 and -3 after serum deprivation for 3 or 4 hours, as shown using Western blotting. Furthermore, serum deprivation-induced apoptosis in a keloid fibroblast line was blocked by a caspase-9 inhibitor (acetyl-Leu-Glu-His-Asp-al), indicating that activation of caspase-9 was necessary for the process of apoptosis in keloid fibroblasts. Although serum deprivation did not significantly change the level of apoptosis protease activating factor-1 in any of the lines, cytochrome c release was detected in cytosolic fractions of the lines after serum deprivation for 3 or 4 hours. These results strongly suggest that keloid fibroblasts are predisposed to apoptosis and cytochrome c release and that caspase-9 activation may underlie regulation of apoptosis in keloid fibroblasts in vivo.

Local administration of hepatocyte growth factor gene enhances the regeneration of dermis in acute incisional wounds.

Hepatocyte growth factor (HGF) has a number of biological activities, e.g., mitogenic, motogenic, antiapoptotic, antifibrous, and morphogenic. It also has angiogenic and angioprotective activities for endothelial cells. The aim of this study was to characterize the role of HGF in wound healing by administering the HGF gene locally to acute incisional skin wounds created on the backs of rats. To create wounds, the backs of Wistar rats were clipped and three 2-cm-long incisional wounds were made deep to the fascia. The wounds contained pannicrus carnosum and were created at intervals of 2 cm. After suturing, the HGF gene was then administered intradermally. Apoptotic cells in wound lesions were identified by TUNEL method as well as by immunological detection of active caspase-3. In the HGF-treated animals, we found almost complete suppression of apoptosis and well-organized wound healing. Histopathological examination revealed that the proliferation of fibroblasts was suppressed and that scar formation was less apparent in the HGF-treated animals compared to the controls. It is thought that administration of the HGF gene immediately after surgery may enhance the healing process through suppressing apoptosis, which occurred in the controls 1 week after suturing the incisional wound. In addition, locally increased HGF expression due to the introduction of the HGF gene to cells around wounds enhances dermal regeneration, possibly by promoting regeneration of dermal tissue, which results in less scarring due to its antifibrotic effect. Thus, HGF supplementation through gene therapy may be an effective strategy for treating wounds, as it increases the regeneration of the dermis to allow for "scarless wound healing."