Master mitotic kinases regulate viral genome delivery during papillomavirus cell entry.

Mitosis induces cellular rearrangements like spindle formation, Golgi fragmentation, and nuclear envelope breakdown. Similar to certain retroviruses, nuclear delivery during entry of human papillomavirus (HPV) genomes is facilitated by mitosis, during which minor capsid protein L2 tethers viral DNA to mitotic chromosomes. However, the mechanism of viral genome delivery and tethering to condensed chromosomes is barely understood. It is unclear, which cellular proteins facilitate this process or how this process is regulated. This work identifies crucial phosphorylations on HPV minor capsid protein L2 occurring at mitosis onset. L2's chromosome binding region (CBR) is sequentially phosphorylated by the master mitotic kinases CDK1 and PLK1. L2 phosphorylation, thus, regulates timely delivery of HPV vDNA to mitotic chromatin during mitosis. In summary, our work demonstrates a crucial role of mitotic kinases for nuclear delivery of viral DNA and provides important insights into the molecular mechanism of pathogen import into the nucleus during mitosis.

Use of Contact Immunotherapy in the Treatment of Skin Diseases Other than Alopecia Areata.

For decades, contact immunotherapy with dinitrochlorobenzene, diphencyprone, and squaric acid dibutylester has played an important role in both clinical practice and scientific research. It is listed as the first-line treatment for extensive alopecia areata and was more recently approved for melanoma treatment as an orphan drug in the USA. Moreover, owing to the relative low cost and safety, topical immunotherapy has also been used in many infectious, neoplastic, and inflammatory dermatological diseases. It is especially valuable in vulnerable groups, for cosmetic/pain sensitive areas, or for multiple lesions. In this review, we summarize the current evidence supporting the use of contact immunotherapy for treatment of skin diseases, from articles collected from PubMed database. Owing to space limitation and already numerous studies focusing on alopecia areata, we include only skin diseases other than alopecia areata. In addition to diseases that have been reported to be treated by contact immunotherapy, the hypothesized mechanism, prognosis prediction, efficacy, and safety of these topical agents are discussed.

Outcomes of Patients With Metastatic Differentiated Thyroid Cancer After Excellent Response to Treatment.

Background: To evaluate the outcomes in differentiated thyroid cancer (DTC) patients who achieved excellent response to initial treatment and developed distant metastasis during follow-up. Methods: Thyroid cancer patients registered in Chang Gung Memorial Hospital thyroid cancer database between January 1979 and December 2019 were assessed. Results: Among 1053 DTC patients with excellent response to initial therapy, 14 (1.3%) patients developed metastatic disease during follow-up, including 6 males and 8 females with median age of 50.2 years [interquartile range (IQR), 39.9-53.7]. Nine (64.3%) patients had papillary cancer, four (28.6%) had follicular cancer, and one (7.1%) had Hürthle cell cancer. Most patients (92.9%) had stage I disease at diagnosis. The sites of metastasis were lung (71.4%), bone (7.1%), mediastinum (7.1%) and multiple sites (14.3%). With a median follow-up of 18.3 years (IQR, 14.8-23.8), 2 patients had disease-specific mortality. The 5- and 10-year disease-specific survival after the diagnosis of distant metastasis was 92% and 74%, respectively. Multiple sites of metastasis was associated with increased risk of mortality (P = 0.022). Conclusions: A small proportion of DTC patients with an excellence response to initial therapy developed distant metastasis during follow-up. Multiple organ distant metastases conferred a worse disease-specific survival.

Preoperative laser reduces silicone oil use in primary diabetic vitrectomy.

AIM: To identify the predictive factors and laser photocoagulation associated with the use of silicone oil as endotamponade during primary diabetic vitrectomy. METHODS: The medical and surgical records of 690 patients (798 eyes) who underwent primary diabetic vitrectomy at a tertiary eye hospital in China from January 2018 to December 2018 were reviewed in this retrospective cohort study. The patients' baseline characteristics and preoperative treatments were recorded. The binary Logistic regression model was used to evaluate the risk factors for the use of silicone oil as endotamponade agent during primary vitrectomy for proliferative diabetic retinopathy (PDR)-related complications. RESULTS: Among 690 patients with mean age of 52.1±10.5y (range: 18-85y), 299/690 (43.3%) were female. The 31.6% of the eyes received preoperative laser treatment, and 72.4% of the eyes received preoperative anti-VEGF adjuvant therapy. Non-clearing vitreous haemorrhage (VH) alone or combined with retinal detachment was the main surgical indication (89.5%) for primary vitrectomy. Silicone oil was used as endotamponade in 313 (39.2%) eyes. Lack of preoperative laser treatment [odds ratio (OR) 0.66, 95% confidence interval (CI): 0.48-0.92; P=0.015] and older age (OR 0.96, 95%CI: 0.95-0.98; P<0.001) were predictors of silicone oil tamponade during primary vitrectomy for PDR. CONCLUSION: The lack of preoperative laser treatment is a significant predictor of silicone oil tamponade during primary vitrectomy for PDR. However, the severity of PDR relevant to silicone oil use should be further evaluated.

A Ran-binding protein facilitates nuclear import of human papillomavirus type 16.

Human papillomaviruses (HPVs) utilize an atypical mode of nuclear import during cell entry. Residing in the Golgi apparatus until mitosis onset, a subviral complex composed of the minor capsid protein L2 and viral DNA (L2/vDNA) is imported into the nucleus after nuclear envelope breakdown by associating with mitotic chromatin. In this complex, L2 plays a crucial role in the interactions with cellular factors that enable delivery and ultimately tethering of the viral genome to mitotic chromatin. To date, the cellular proteins facilitating these steps remain unknown. Here, we addressed which cellular proteins may be required for this process. Using label-free mass spectrometry, biochemical assays, microscopy, and functional virological assays, we discovered that L2 engages a hitherto unknown protein complex of Ran-binding protein 10 (RanBP10), karyopherin alpha2 (KPNA2), and dynein light chain DYNLT3 to facilitate transport towards mitotic chromatin. Thus, our study not only identifies novel cellular interactors and mechanism that facilitate a poorly understood step in HPV entry, but also a novel cellular transport complex.

Novel roles of folic acid as redox regulator: Modulation of reactive oxygen species sinker protein expression and maintenance of mitochondrial redox homeostasis on hepatocellular carcinoma.

We provide herein several lines of evidence to substantiate that folic acid (or folate) is a micronutrient capable of functioning as a novel redox regulator on hepatocellular carcinoma. First, we uncovered that folate deficiency could profoundly downregulate two prominent anti-apoptotic effectors including survivin and glucose-regulated protein-78. Silencing of either survivin or glucose-regulated protein-78 via small interfering RNA interfering technique established that both effectors could serve as reactive oxygen species sinker proteins. Second, folate deficiency-triggered oxidative-nitrosative stress could strongly induce endoplasmic reticulum stress that in turn could provoke cellular glutathione depletion through the modulation of the following two crucial events: (1) folate deficiency could strongly inhibit Bcl-2 expression leading to severe suppression of the mitochondrial glutathione pool and (2) folate deficiency could also profoundly inhibit two key enzymes that governing cellular glutathione redox regulation including γ-glutamylcysteinyl synthetase heavy chain, a catalytic enzyme for glutathione biosynthesis, and mitochondrial isocitrate dehydrogenase 2, an enzyme responsible for providing nicotinamide adenine dinucleotide phosphate necessary for regenerating oxidized glutathione disulfide back to glutathione via mitochondrial glutathione reductase. Collectively, we add to the literature new data to strengthen the notion that folate is an essential micronutrient that confers a novel role to combat reactive oxygen species insults and thus serves as a redox regulator via upregulating reactive oxygen species sinker proteins and averting mitochondrial glutathione depletion through proper maintenance of redox homeostasis via positively regulating glutathione biosynthesis, glutathione transporting system, and mitochondrial glutathione recycling process.

A central region in the minor capsid protein of papillomaviruses facilitates viral genome tethering and membrane penetration for mitotic nuclear entry.

Incoming papillomaviruses (PVs) depend on mitotic nuclear envelope breakdown to gain initial access to the nucleus for viral transcription and replication. In our previous work, we hypothesized that the minor capsid protein L2 of PVs tethers the incoming vDNA to mitotic chromosomes to direct them into the nascent nuclei. To re-evaluate how dynamic L2 recruitment to cellular chromosomes occurs specifically during prometaphase, we developed a quantitative, microscopy-based assay for measuring the degree of chromosome recruitment of L2-EGFP. Analyzing various HPV16 L2 truncation-mutants revealed a central chromosome-binding region (CBR) of 147 amino acids that confers binding to mitotic chromosomes. Specific mutations of conserved motifs (IVAL286AAAA, RR302/5AA, and RTR313EEE) within the CBR interfered with chromosomal binding. Moreover, assembly-competent HPV16 containing the chromosome-binding deficient L2(RTR313EEE) or L2(IVAL286AAAA) were inhibited for infection despite their ability to be transported to intracellular compartments. Since vDNA and L2 were not associated with mitotic chromosomes either, the infectivity was likely impaired by a defect in tethering of the vDNA to mitotic chromosomes. However, L2 mutations that abrogated chromatin association also compromised translocation of L2 across membranes of intracellular organelles. Thus, chromatin recruitment of L2 may in itself be a requirement for successful penetration of the limiting membrane thereby linking both processes mechanistically. Furthermore, we demonstrate that the association of L2 with mitotic chromosomes is conserved among the alpha, beta, gamma, and iota genera of Papillomaviridae. However, different binding patterns point to a certain variance amongst the different genera. Overall, our data suggest a common strategy among various PVs, in which a central region of L2 mediates tethering of vDNA to mitotic chromosomes during cell division thereby coordinating membrane translocation and delivery to daughter nuclei.

Cisplatin-induced regulation of signal transduction pathways and transcription factors in p53-mutated subclone variants of hepatoma cells: Potential application for therapeutic targeting.

Cisplatin is commonly recognized as a DNA-damaging drug; however, its versatile antitumor effects have been demonstrated to extend beyond this narrow functional attribute. The present study determined how cisplatin regulates alternative pathways and transcription factors to exert its additional antitumor actions. Cisplatin was observed to be able to trigger an endoplasmic reticulum stress response through aggravated nitrosative stress coupled to perturbed mitochondrial calcium (Ca2+) homeostasis, which substantially downregulated glucose-regulated protein (GRP) 78 expression by suppressing the cleavage of activating transcription factor (ATF) 6α (90 kDa) to its active 50 kDa subunit. Concomitantly, the ATF4-ATF3-C/emopamil binding protein homologous protein axis was activated by cisplatin, which triggered cellular glutathione (GSH) depletion by strongly inhibiting γ-glutamylcysteine synthetase heavy chain (γ-GCSh), a key enzyme in GSH biosynthesis. The present study also demonstrated that cisplatin substantially inhibited ß-catenin, causing a marked downregulation of survivin and B-cell lymphoma (Bcl)-2. Taken together, the present results uncovered a novel mechanism of cisplatin that could simultaneously trigger the inhibition of three prominent antiapoptotic effector molecules (Bcl-2, survivin and GRP78) and effectively promote GSH depletion by inhibiting γ-GCSh. These newly discovered functional attributes of cisplatin can provide an avenue for novel combined therapeutic strategies to kill hepatocellular carcinoma cells effectively.

Paclitaxel pretreatment overcomes hypoxia inducible factor-1α-induced radioresistance acquisition of human hepatoma and lung adenocarcinoma cells.

AIMS: This study delineated the mechanisms of paclitaxel (PTX) assistance in overcoming radioresistance in hepatoma and human lung adenocarcinoma (HLAC) cells. MAIN METHODS: The TUNEL assay was used as an index of radiosensitivity, and the MTT assay assessed the efficacy of various combined PTX/RT treatments. The efficacy of PTX disruptions of hypoxia-inducible factor-1 alpha (HIF-1α) was assessed using Western blotting. KEY FINDINGS: Normoxically overexpressed HIF-1α in hepatoma J5 cells was mechanistically linked to activation of the bFGF/PI3K/Akt pathway because the viability of these cells was strongly inhibited by either Akt inhibitors or an HIF-1α inhibitor. All of the cell lines used were extremely sensitive to PTX, and these effects also correlated excellently with HIF-1α suppression. We designed five combined radiation-PTX protocols of varying dose duration and treatment sequences against CL1-1 cells based on the gathered data. Pretreatment of CL1-1 cells with PTX (100nM) for 24h before irradiation (2.5Gy) was the best combined protocol to achieve maximum radiosensitizing effects. SIGNIFICANCE: Our data clearly indicate that PTX pretreatment is an effective radiosensitizing procedure against HIF-1α-expressing hepatoma and HLAC cells, which are constitutively endowed with radioresistance.

Maintenance of Epstein-Barr Virus Latent Status by a Novel Mechanism, Latent Membrane Protein 1-Induced Interleukin-32, via the Protein Kinase Cδ Pathway.

UNLABELLED: Epstein-Barr virus (EBV), an oncogenic herpesvirus, has the potential to immortalize primary B cells into lymphoblastoid cell lines (LCLs) in vitro. During immortalization, several EBV products induce cytokines or chemokines, and most of these are required for the proliferation of LCLs. Interleukin-32 (IL-32), a recently discovered proinflammatory cytokine, is upregulated after EBV infection, and this upregulation is detectable in all LCLs tested. EBV latent membrane protein 1 (LMP1) is responsible for inducing IL-32 expression at the mRNA and protein levels. Mechanistically, we showed that this LMP1 induction is provided by the p65 subunit of NF-κB, which binds to and activates the IL-32 promoter. Furthermore, the short hairpin RNA (shRNA)-mediated depletion of endogenous LMP1 and p65 in LCLs suppressed IL-32 expression, further suggesting that LMP1 is the key factor that stimulates IL-32 in LCLs via the NF-κB p65 pathway. Functionally, knockdown of IL-32 in LCLs elicits viral reactivation and affects cytokine expression, but it has no impact on cell proliferation and apoptosis. Of note, we reveal the mechanism whereby IL-32 is involved in the maintenance of EBV viral latency by inactivation of Zta promoter activity. This atypical cytoplasmic IL-32 hijacks the Zta activator protein kinase Cδ (PKCδ) and inhibits its translocation from the cytoplasm to the nucleus, where PKCδ binds to the Zta promoter and activates lytic cycle progression. These novel findings reveal that IL-32 is involved in the maintenance of EBV latency in LCLs. This finding may provide new information to explain how EBV maintains latency, in addition to viral chromatin structure and epigenetic modification. IMPORTANCE: EBV persists in two states, latency and lytic replication, which is a unique characteristic of human infections. So far, little is known about how herpesviruses maintain latency in particular tissues or cell types. EBV is an excellent model to study this question because more than 90% of people are latently infected. EBV can immortalize primary B cells into lymphoblastoid cell lines in vitro. Expression of IL-32, a novel atypical cytoplasmic proinflammatory cytokine, increased after infection. The expression of IL-32 was controlled by LMP1. In investigating the regulatory mechanism, we demonstrated that the p65 subunit of NF-κB is required for this upregulation. Of note, the important biological activity of IL-32 was to trap protein kinase Cδ in the cytoplasm and prevent it from binding to the Zta promoter, which is the key event for EBV reaction. So, the expression of LMP1-induced IL-32 plays a role in the maintenance of EBV latency.

Retinal vein occlusion in retinal racemose hemangioma: a case report and literature review of ocular complications in this rare retinal vascular disorder.

BACKGROUD: Retinal racemose hemangioma (RRH) is a rare congenital disorder that often co-occurs with other ocular complications. In this study, we present a case of RRH complicated with retinal vein obstruction in three branches and provide a review of ocular complications and associations with RRH. CASE PRESENTATION: One case of RRH is presented. Fundus examination, fluorescein angiography (FFA) and optical coherence tomography (OCT) of the patient identified Group 3 RRH complicated with retinal vein occlusions in the superotemporal, inferotemporal, and inferonasal branches. Macular edema, which causes visual impairment, was detected. A brief literature review was also presented. The PubMed database was searched for RRH or related keywords to find reports of ocular complications or associations published on or before Dec. 31, 2013. A total of 140 papers describing167 RRH cases were found. The mean age of diagnosis was 22.97 years. Ocular complications were mentioned in 32 (19.16%) cases. Retinal vein occlusion (46.88%) was the major ocular complication in RRH, followed by hemorrhage (34.38%). Eight (4.79%) cases were associated with other ocular diseases such as Sturge-Weber syndrome , Morning glory disc anomaly and macroaneurysm. CONCLUSIONS: Although RRH is a relatively non-progressive condition, its complications may lead to vision loss and should be treated in time.

Periodic Si nanopillar arrays by anodic aluminum oxide template and catalytic etching for broadband and omnidirectional light harvesting.

Large-area, periodic Si nanopillar arrays (NPAs) with the periodicity of 100 nm and the diameter of 60 nm were fabricated by metal-assisted chemical etching with anodic aluminum oxide as a patterning mask. The 100-nm-periodicity NPAs serve an antireflection function especially at the wavelengths of 200~400 nm, where the reflectance is decreased to be almost tenth of the value of the polished Si (from 62.9% to 7.9%). These NPAs show very low reflectance for broadband wavelengths and omnidirectional light incidence, attributed to the small periodicity and the stepped refractive index of NPA layers. The experimental results are confirmed by theoretical calculations. Raman scattering intensity was also found to be significantly increased with Si NPAs. The introduction of this industrial-scale self-assembly methodology for light

Heat shock factor 1 is a transcription factor of Fas gene.

In mammalian cells, stress-induced expression of heat shock protein is controlled by heat shock factor 1 (HSF1). However, HSF1 functions as a regulator of additional genes. In this study, we observed that heat treatment effectively induced expression of Fas. Using bioinformatics, a high affinity and functional HSF1-binding element within the -1996/-1985 oligonucleotide of the 5'-flanking region of the Fas gene was found, and was determined by electrophoretic mobility shift assay and chromatin immunoprecipitation assay. Exogenous expression of a constitutively activative HSF1, induced Fas gene transcription and protein synthesis in the absence of heat stress. Moreover, RNA interference-mediated HSF1 gene-silencing attenuated Fas expression in a heat-induced model. Our results suggested that HSF1 is an important transcription factor of Fas gene.

Outcome analysis of patients with craniofacial resection: Hong Kong experience.

BACKGROUND: The aim of this study was to evaluate the clinical outcome of patients who underwent craniofacial resection for tumour in the anterior skull base at the University of Hong Kong Medical Centre, Queen Mary Hospital, Hong Kong between January 1993 and June 2003. METHODS: A retrospective review was conducted. The duration of follow up ranged from 1.7 to 119 months (median, 41.8 months). The setting was a tertiary referral centre. Thirty-nine patients, 23 males and 16 females, aged 8-79 years were included. Thirty-four patients had malignant tumour and five patients had benign pathology. The tumour pathology was diversified. Nineteen patients had no treatment before the surgery, and the remaining 20 patients had received surgery, radiotherapy or combined treatment before resection. Patients were given postoperative irradiation and/or chemotherapy depending on the pathology of the tumour and the extent of the disease and clearance at the time of surgery. RESULTS: There was no operative mortality. Complications occurred in 10 patients and among them, three required reoperation. The other complications were managed successfully with conservative measures. The 5-year actuarial disease-free survival for patients with benign and malignant pathology was 100 and 77.6%, respectively. For patients with malignant pathology, 5-year actuarial disease-free survival was 90% when the resection margin was negative at surgery. However, the survival dropped to 53.6% when the resection margin was involved macroscopically. CONCLUSION: Craniofacial resection was an appropriate surgical approach with acceptable morbidity in selected patients with tumour located at the anterior skull base. Complete excision of malignant tumour could achieve 90% 5-year disease-free actuarial survival.

Use of 1,320 nm Nd:YAG laser for wrinkle reduction and the treatment of atrophic acne scarring in Asians.

BACKGROUND AND OBJECTIVES: The role of 1,320 Nd:YAG in non-ablative skin rejuvenation in Asians is has not been established. Furthermore, no study has investigated the effectiveness of 1,320 Nd:YAG laser in the treatment of atrophic scarring in Asians. The objective of our study was to investigate the effectiveness of 1,320 Nd:YAG laser in wrinkle reduction and the treatment of atrophic acne scarring in Asians. STUDY DESIGN/MATERIALS AND METHODS: Twenty-seven female patients were included in the study: seven were treated for acne scarring and the others for wrinkle reduction. A 1,320 nm Nd:YAG laser (Cooltouch II, Roseville, CA) was used to treat both the cheeks and forehead for the patients with wrinkles, and both cheeks only for patients with atrophic acne scarring. All patients received treatment in the post-auricular areas. A spot size of 10 mm was used, and three passes were performed (two pre-cooling and one post-cooling). Patients were treated monthly for 6 months. All patients were subjectively assessed before and after their last treatment sessions using a structured questionnaire, and objectively assessed by the use of clinical photographs for by independent observers. A cutometer was used to assess viscoelasticity, and biopsies were taken at the post-auricular site for assessment by a pathologist. RESULTS: The overall degree of patients' satisfaction was rated as 4.9 (range 0-9.8) for wrinkle reduction and 4 (range 0-10) for acne scarring. In terms of objective assessment by independent observers, the degree of improvement was considered to be mild or no change in most cases. The independent pathologist who assessed the degree of improvement in terms of increased collagen production detected no change in 8 patients, mild improvement in 9, and moderate improvement in 10. There was also improvement in term of epidermal thickness in 13 cases. Assessment by viscoelasticity indicated a significant degree of improvement in most parameters in both groups of patients. Blistering occurred in five cases, all in the central facial areas, and post-inflammatory hyperpigmentation occurred in three cases. All cases of PIH resolved after the use of 4% hydroquinoine. There was no scarring or hypopigmentation. CONCLUSIONS: The 1,320 nm Nd:YAG laser is effective for wrinkle reduction and atrophic acne scar improvement, but to further enhance the clinical outcome a combination approach with another device such as IPL and a surgical technique such as subcision is necessary.

Microvascular reconstruction of the hepatic artery in live donor liver transplantation: experience across a decade.

HYPOTHESIS: Hepatic arterial anastomosis by means of a microvascular technique can be performed with a high success rate in live donor liver transplantation in adult recipients. DESIGN: Retrospective analysis of data collected prospectively. SETTING: Tertiary referral center. PATIENTS: From September 28, 1993, through December 23, 2002, 28 children received left lateral segment grafts (n = 23) or left lobe grafts (n = 5), and 124 adults received left lobe (n = 6) or right lobe (n = 118) grafts. Microvascular technique was used for hepatic arterial anastomosis. Attention was paid to exposure, orientation of the axis of arterial ends, and matching of size. Long microinstruments were used for arterial ends deeply located inside an adult's abdominal cavity. An average of 16 stitches was used for a vessel anastomosis 3 mm in diameter. INTERVENTIONS: Intraoperative and postoperative Doppler ultrasonography were performed. MAIN OUTCOME MEASURES: Hepatic artery thrombosis rate. RESULTS: All hepatic arterial anastomoses were patent after reconstruction. Complications occurred in 3 patients. They had hepatic arterial thrombosis at 19 days, 25 days, and 3 months after liver transplantation. The overall complication rate was 2%. CONCLUSION: With attention to exposure, appropriate instruments, and experience, a low complication rate of 2% can be achieved in hepatic arterial anastomosis by using a microvascular technique, even in adult patients with the liver graft situated deeply in the abdominal cavity.

Salvage of recurrent head and neck squamous cell carcinoma after primary curative surgery.

PURPOSE: The efficacy of salvage treatment of recurrent head and neck squamous cell carcinomas (HNSCC) after primary curative surgery was evaluated. METHODS: The management outcome of 377 patients who had recurrent squamous cell carcinoma of oral cavity, oropharynx, hypopharynx, and larynx after primary curative surgery was reviewed. RESULTS: The surgical salvage rates of recurrence were 29% local, 30% tracheostomal, 56% unilateral nodal recurrence of previously undissected neck, 32% of unilateral neck recurrence after prior neck dissection, and 11% lung metastasis. The 5-year tumor-free actuarial survival rates of those patients who received surgical salvage was 35% for local recurrence, 32% for unilateral nodal recurrence of the previously undissected neck, and 18% for nodal recurrence of the previously dissected neck. One patient of six with tracheostomal recurrence salvaged with surgery and one patient of six with lung metastasis salvaged with lobectomy survived without tumor at 5 years. There was no 5-year survivor of all patients salvaged with other nonsurgical methods. The mean survival of patients without surgical salvage was 6 months. CONCLUSIONS: There was a moderate chance of cure after surgical salvage of locoregional recurrent HNSCC. Surgical salvage was, however, only feasible for early recurrent tumor. Close follow-up surveillance of early recurrence is essential after primary treatment of patients.

[Inhibition of tongue cancer development in nude mice transfected with adenovirus carrying human endostatin gene].

BACKGROUND & OBJECTIVE: The squamous cell carcinoma of tongue is one of the most common malignant tumors of oral cavity. Surgical therapy is now the mainstay of combined treatment for tongue squamous cell Carcinoma with chemotherapy and radiotherapy. The overall 5-year survival rate was about 50%. The antiangiogenesis therapy has become a new approach of the treatment of tongue carcinoma. This paper was designed to study the characteristics of endostatin expression in tongue cancer cell line (Tca8113), human embryonic epithelial cell line (ECV) and the inhibition of carcinogenesis in nude mice, xenografted with Tca8113, after transfected with recombined adenovirus (Ad/hEnd) which was cloned with human endostatin gene in EI mutated region. METHODS: (1) To determine the expression and distribution of endostatin in Tca and ECV cells transfected with Ad/hEnd using immunohistochemistry. To determine the endostatin in supernatants of Tca cells transfected with Ad/hEnd using ELISA method. To examine the characteristics of endostatin gene expression in Tca8113 and ECV cells by Western blot analysis. (2) To determine the inhibition rate of proliferation and apoptosis rate of ECV cells by WST-1 test and flow cytometry (FCM), respectively.(3) To observe the inhibition of tumor growth in xenografted nude mice with Tca8113 cells by Ad/hEnd administration. RESULTS: (1) Immunohistochemistry detection indicated that the endostatin was expressed in cytoplasm of Tca8113 cells and ECV cells transfected with Ad/hEnd. Endostatin expression in the supernatant was dose-dependent with the highest to 597 ng/ml. The expression of endostatin in Tca cells was detectable from 1 day to 7 day. Ad/hEnd inhibited ECV cell growth in dose-dependent manner. (2) FCM showed that Ad/hEnd arrested ECV cells in S and G(2) phase and induced apoptosis.(3) The tumor growth curve showed that Ad/hEnd significantly repressed xenograft tumor growth with Tca cell in nude mice; the inhibition rate on Ad/hEnd administrated groups was 45.8% in the 3rd week. CONCLUSION: Ad/hEnd expressed efficiently in Tca8113 and ECV cells. Ad/hEnd can change the cell cycle distribution of ECV cells and induce apoptosis and inhibit proliferation of ECV cells. Ad/hEnd could inhibit the growth of tongue carcinoma in xenograft nude mice.

Role of skin cooling in improving patient tolerability of Q-switched Alexandrite (QS Alex) laser in nevus of Ota treatment.

BACKGROUND: The development of a high-energy laser device with very short pulse duration has revolutionized the treatment of nevus of Ota. Nevertheless, patients still suffer from complications that range from the pain and swelling that occur immediately after laser surgery to post-operative pigmentary changes and scarring. However, the simultaneous use of skin surface cooling and laser surgery may limit the damage of non-target tissue. OBJECTIVES: To assess whether epidermal cooling reduces the pain and swelling that commonly occur after laser treatment for nevus of Ota. STUDY DESIGN/PATIENTS AND METHODS: Thirty-seven patients with nevus of Ota were recruited from a dermatology outpatient clinic. Before treatment, the research nurse used an ink pen to divide the lesions into two halves. Half of each lesion was treated with a Q-switched Alex laser system that had a cool sapphire plate in contact as a mean of skin cooling. The other half was treated with the same laser, but with the cooling device switched off. Immediately after treatment, and again 1 week later, the patients answered a questionnaire, which assessed the symptoms that are associated with laser surgery. Dose assessment was performed in each half of the nevus to obtain the optimal fluence that could be used for the treatment of nevus of Ota before the entire half was treated. RESULTS: There was no difference in terms of the optimal fluence that was used, but in terms of immediate pain the patients associated the side that was treated with the cooling plate with a significantly lesser degree of pain than the non-cooled side (P = 0.001). Eighty two percent of the patients preferred the cooled side to the non-cooled side. CONCLUSIONS: Pre- and post-operative skin cooling is effective in improving the tolerability of nevus of Ota patients to Q-switched laser treatment. However, the use of cooling during laser treatment of nevus of Ota did not allow the use of higher fluence.

Prognostic factors of clinically stage I and II oral tongue carcinoma-A comparative study of stage, thickness, shape, growth pattern, invasive front malignancy grading, Martinez-Gimeno score, and pathologic features.

PURPOSE: This study aims at evaluation of the different prognostic models, including stage, tumor thickness, shape, malignancy grading of tumor invasive front, Martinez-Gimeno score, and pathologic features in the prediction of subclinical nodal metastasis, local recurrence, and survival of early T1 and T2 oral tongue squamous cell carcinoma. The results will have important implication for the management of patients. PATIENTS AND METHODS: Seventy-two clinically T1 and T2 glossectomy specimens of oral tongue carcinoma were serially sectioned in 3-mm thickness for the evaluation of various pathologic features. The prognostic value in the prediction of subclinical nodal metastasis, local recurrence, and survival of different models were compared. RESULTS: Among all the tumor parameters and predictive models being evaluated, tumor thickness was the only significant factor that had significant predictive value for subclinical nodal metastasis, local recurrence, and survival. With the use of 3-mm and 9-mm division, tumor of up to 3-mm thickness has 8% subclinical nodal metastasis, 0% local recurrence, and 100% 5-year actuarial disease-free survival; tumor thickness of more than 3 mm and up to 9 mm had 44% subclinical nodal metastasis, 7% local recurrence, and 76% 5-year actuarial disease-free survival; tumor of more than 9 mm had 53% subclinical nodal metastasis, 24% local recurrence, and 66% 5-year actuarial disease-free survival. CONCLUSIONS: Tumor thickness should be considered in the management planning of patients with early oral tongue carcinoma.