[Study on self-microemulsifying membrane controlled-release drop pill of hawthorn leaves flavonoids].

To prepare the hawthorn leaves flavonoids self-microemulsifying membrane controlled-release coated drop pill, and to study its release rate in vitro and pharmacokinetics study in vivo. In order to improve the dissolution of hawthorn leaves flavonoids, self-microemulsifying technology was used to prepare the hawthorn leaves flavonoids self-microemulsion. Hawthorn leaves flavonoids self-microemulsifying drop pill was prepared with the PEG 6000. Studies were made on the in vitro release of flavonoids from hawthorn leaves self-micro-emulsifying membrane-moderated coated drop pills and the in vivo pharmacokinetic in rats. The prescription of flavonoids from hawthorn leaves self-micro-emulsifying drop pills was 0.25 g of flavonoids from hawthorn leaves, 0.25 g of iodophenyl maleimide, 0.375 g of polyethylene glycol 400, 0.375 g of cremophor RH 40 and 2 g of polyethylene glycol 6000. The optimized prescription was 4 g of ethyl cellulose 20, 0.64 g of polyethylene glycol 400, 1.8 g of diethyl phthalate, and the weight of coating materials increased by 3.5%. Flavonoids from hawthorn leaves self-micro-emulsifying membrane-moderated coated drop pills complied with the design of sustained-release in 12 h in terms of in vitro release and in vivo pharmacokinetic parameters in rats, and its bioavailability was 2.47 times of quick-release drop pills. Slightly soluble flavonoids from hawthorn leaves could be made into sustained-release preparations by the self-micro-emulsifying and coating technology.

Boosting Upconversion Efficiency in Optically Inert Shelled Structures with Electroactive Membrane through Electron Donation.

This study innovatively addresses challenges in enhancing upconversion efficiency in lanthanide-based nanoparticles (UCNPs) by exploiting Shewanella oneidensis MR-1, a microorganism capable of extracellular electron transfer. Electroactive membranes, rich in c-type cytochromes, are extracted from bacteria and integrated into membrane-integrated liposomes (MILs), encapsulating core-shelled UCNPs with an optically inactive shell, forming UCNP@MIL constructs. The electroactive membrane, tailored to donate electrons through the inert shell, independently boosts upconversion emission under near-infrared excitation (980 or 1550 nm), bypassing ligand-sensitized UCNPs. The optically inactive shell restricts energy migration, emphasizing electroactive membrane electron donation. Density functional theory calculations elucidate efficient electron transfer due to the electroactive membrane hemes' highest occupied molecular orbital being higher than the valence band maximum of the optically inactive shell, crucial for enhancing energy transfer to emitter ions. The introduction of a SiO2 insulator coating diminishes light enhancement, underscoring the importance of unimpeded electron transfer. Luminescence enhancement remains resilient to variations in emitter or sensitizing ions, highlighting the robustness of the electron transfer-induced phenomenon. However, altering the inert shell material diminishes enhancement, emphasizing the role of electron transfer. This methodology holds significant promise for diverse biological applications. UCNP@MIL offers an advantage in cellular uptake, which proves beneficial for cell imaging.

Suppression of tongue squamous cell carcinoma growth by inhibition of Jagged1 in vitro and in vivo.

BACKGROUND: The changes in Notch signaling are closely related to the occurrence and development of many cancers. We have investigated Notch signaling receptor and its ligand expressions in TSCC cell lines, tissues and its significance. We clarified Notch signaling pathway in TSCC and its mechanism. We regulated Notch signaling pathway of tumor cells, thereby inhibiting tumor cell proliferation and differentiation. METHODS: We detected Jagged1 protein and mRNA expression levels in specimens (tongue cancer and adjacent tissues) from 74 patients with tongue cancer and in TSCC cell line. The Jagged1-targeted lentiviral vector RNAi system was constructed, and its suppressive effects on the proliferation and invasion of tongue carcinoma cells in in vivo and ex vivo were determined. RESULTS: Jagged1 was expressed in tongue squamous cell cancer tissues and cell line, but there were differences in its expression. Jagged1 was knocked down and the tumor growth was inhibited accompanying cell cycle changes. Animal studies also showed that the tumor growth was inhibited. CONCLUSIONS: Jagged1 may be involved in the differentiation and proliferation of tongue cancer. Targeting Jagged1 RNA interference lentiviral vector can effectively lower Jagged1 mRNA and protein expression levels of Tca8113 cells, thereby preventing the proliferation of TSCC cells. Jagged1 is expected to be a promising new target for curing tongue cancer. In-depth study of the interaction between Jagged1 and other molecules of Notch signaling pathway in the process of carcinogenesis has important theoretical guidance and clinical significance in revealing the mechanism of Jagged1 and its application in the therapy for tongue cancer.

The expression and significance of MRP1, LRP, TOPOIIß, and BCL2 in tongue squamous cell carcinoma.

BACKGROUND: Multiple drug resistance protein 1 (MRP1), lung resistance protein (LRP), topoisomerase IIß (TOPOIIß) and B-cell lymphoma 2 (BCL2) are well known in the development of drug resistance in cancer cells. The aim of this study was to evaluate the relationship between them and the clinicopathological features, their expression differences between tumor tissue and experimental drug-resistant model in tongue carcinoma. MATERIALS AND METHODS: Multiple drug resistance protein 1, LRP, TOPOIIß, and BCL2 expression was examined by immunohistochemistry in specimens from radical surgeries of 65 patients with tongue carcinoma. A cisplatin-resistance cell line, SCC-15/cisplatin, was established from a cisplatin-sensitive cell line, SCC-15. A MTT-based method was used to analyze drug potencies. Immunofluorescence was used to detect protein expression in both cell lines. Western blot was used to compare the protein expressions in specimens and SCC-15/cisplatin cells. RESULTS: We found higher expression of MRP1, LRP, and BCL2 and lower expression of TOPOIIß in tongue carcinoma compared with adjacent non-neoplastic tongue tissues (P < 0.05). In addition, MRP1 and TopoIIß expression were significantly associated with clinical stage, lymph node metastasis and histologic grade, and LRP was significantly associated with histologic grade in the samples (P < 0.05). Finally, Western blot showed that higher expressions of MRP1, LRP, and BCL2 and lower expression of TopoIIß were observed in SCC-15/cisplatin cells than in clinical samples. CONCLUSION: Our results suggest that the high expressions of MRP1, LRP, and BCL2 and low expression of TOPOIIß in patients with tongue carcinoma indicates that intrinsic drug resistance may exist in tongue carcinoma, and is associated with tumor differentiation and cisplatin resistance in tongue carcinoma.

Cephalometric analysis of craniofacial malformations in newborn mice with cleft palate induced by retinoic acid.

OBJECTIVE: To determine changes on craniofacial growth morphometrically in newborn mice with cleft palate induced by retinoic acid. DESIGN, SETTING, PARTICIPANTS, INTERVENTIONS: Gestation day 10 or 12 pregnant female C57BL/6N mice were given a single dose of all-trans retinoic acid (atRA) by gastric intubations via oral gavage. Sixty newborn mice with cleft palate (CP), 52 without CP from the experimental group, and 30 without CP from the control group were collected, and lateral cephalograms were taken of all of the mice. MAIN OUTCOME MEASURES: Cephalometric analysis of the craniofacial skeleton was performed by means of a personal computer. RESULTS: Inhibition of craniofacial growth was found in the experimental groups but not in the control groups. In the maxillary bone and mandible, the amount of growth was significantly reduced. CONCLUSIONS: These results suggest that craniofacial growth is inhibited in newborn mice with cleft palate induced by retinoic acid.

Retinoic acid inhibits osteogenic differentiation of mouse embryonic palate mesenchymal cells.

BACKGROUND: All-trans-retinoic acid (ATRA), a known teratogenic factor affecting the development of cleft palate, has been shown to adversely affect craniofacial development. In the present study, we evaluated the effects of ATRA on the osteo-/adipogenic differentiation of mouse embryonic palate mesenchymal (MEPM) cells, which served as a valid model system for investigating the mechanisms regulating osteogenesis during palatogenesis. METHODS: MEPM cells were derived from gestational day 13 C57BL/6N mouse embryos and induced to differentiate in the presence or absence of ATRA in either osteogenic medium (OM) or control medium (CM). RESULTS: Alkaline phosphatase (ALP) activity assays, von Kossa staining, and RT-PCR assays confirmed that MEPM cells underwent osteogenic differentiation when cultured in OM. Although ATRA induced ALP activity and lipid accumulation in MEPM cells, it failed to induce matrix mineralization and osteoblastic gene expression. BMPR-IB and Smad5 mRNA levels increased significantly in cells cultured in OM and declined following treatment with ATRA, whereas the expression of the BMPR-IA mRNA was up-regulated by ATRA. CONCLUSIONS: In conclusion, our results suggested that ATRA and the BMP signaling pathway cooperate to inhibit osteogenesis and promote adipogenesis of MEPM cells.

Survivin shRNA induces caspase-3-dependent apoptosis and enhances cisplatin sensitivity in squamous cell carcinoma of the tongue.

Survivin is a member of the inhibitor of apoptosis protein (IAP) family; it is overexpressed in most cancer tissues and induces resistance to chemotherapy. In this study, we investigated whether a short hairpin RNA (shRNA) targeting survivin can induce apoptosis and enhance chemosensitivity to cisplatin in squamous cell carcinoma of the tongue. Results showed that chemosensitivity to cisplatin was surviving dependent in three cell lines (Tca8113, Bca885, and MCF7); higher survivin mRNA expression levels were associated with lower sensitivity to cisplatin. A plasmid-containing survivin shRNA was constructed and transfected into cell line Tca8113. Survivin shRNA inhibited expression of survivin mRNA and protein (63% and 65% inhibition, respectively), significantly inhibited cell proliferation, and enhanced chemosensitivity to cisplatin (p < 0.05). Apoptosis and caspase-3 activity were induced when cells were treated with survivin shRNA and/or cisplatin. Survivin shRNA induced caspase-3-dependent apoptosis and enhanced chemosensitivity to cisplatin in these tongue squamous cell carcinoma cell lines.

The role of RANKL and MMP-9 in the bone resorption caused by ameloblastoma.

BACKGROUND: Ameloblastoma, a common odontogenic tumor located in jaws, generally leads to severe damage to patient's complexion and masticatory function. To expand in jaws, ameloblastoma must have a mechanism of resorbing the surrounding bone. Our objective was to explore the bone-resorption mechanism of ameloblastoma by observing the role of Receptor activator of nuclear factor kappa B ligand (RANKL) and matrix metalloproteinase-9 (MMP-9) in the bone-resorption process. METHODS: In the study, the expression of RANKL and MMP-9 in ameloblastoma was detected using immunohistochemistry (IHC) and RT-PCR. Then, co-culture system of ameloblastoma cells and bone marrow cells from neonatal rabbit was erected to observe the potential of ameloblastoma cells to induce osteoclastogenesis. Finally, the induced osteoclasts were used for in vitro bone-resorption assay. In the co-culture system and the bone-resorption assay, the selective inhibitor of RANKL and MMP-9, osteoprotegerin (OPG) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were, respectively, used for observing the role of RANKL and MMP-9. RESULTS: The expression of RANKL and MMP-9 in ameloblastoma was confirmed. Ameloblastoma cells were found to induce bone marrow cells from neonatal rabbit differentiate into osteoclasts with bone-resorption activity. In addition, OPG was found to, respectively, have markedly inhibitory effect on osteoclastogenesis (P<0.01), and slightly inhibitory action on bone resorption (P<0.05). CONCLUSIONS: Ameloblastoma cells had the potential to induce osteoclastogenesis. Moreover, RANKL played an essential role in the in vitro osteoclast formation and bone resorption induced by ameloblastoma cells.

Expression and role of metalloproteinase-2 and endogenous tissue regulator in ameloblastoma.

BACKGROUND: Ameloblastoma is a frequently encountered odontogenic benign tumor characterized by local invasiveness and high risk of recurrence. Matrix metalloproteinase (MMP)-2 can degrade type IV collagen, one of the major components in the basement membrane, resulting in the promotion of tumor invasion, whereas it is under the influence of an activator, membrane type 1-MMP-14 and the tissue inhibitor of metalloproteases (TIMP)-2. The aim of this study was to investigate combinatorial role played by MMP-2, TIMP-2 and MMP-14 in ameloblastoma. METHODS: Transcriptional expression of MMP-2, TIMP-2 and MMP-14 in tissue extracts of 42 ameloblastomas and 10 dental follicles was detected using reverse transcription-polymerase chain reaction, and compared difference in clinical type and local recurrence of ameloblastoma. RESULTS: MMP-2, TIMP-2 and MMP-14 mRNA were detected in all investigated ameloblastoma tissues. While MMP-2 mRNA was only expressed in eight of 10 odontotheca tissues and TIMP- 2 and MMP-14 were not found in all odontotheca tissues. The mRNA expression of MMP-2, TIMP-2 and MMP-14 were significantly higher in ameloblastoma tissues than in odontotheca tissues. The mRNA levels of TIMP-2 and MMP-14 were significantly higher in recurrent and solid/multicystic ameloblastoma tissues than in primary and unicystic ameloblastoma tissues respectively. CONCLUSIONS: Of the MMP-2/TIMP-2/MMP-14 complex, a high expression of MMP-2, TIMP-2 and MMP-14 mRNA levels may contribute to the local invasive characteristics of ameloblastoma, whereas the local invasive capacity of ameloblastoma is more likely related to a high transcriptional levels of TIMP-2 and MMP-14.

Quality of life of patients with tongue cancer 1 year after surgery.

PURPOSE: To study the changes and factors affecting the quality of life (QOL) of patients with tongue cancer 1 year after primary surgery. PATIENTS AND METHODS: A total of 289 consecutive patients with tongue cancer who had undergone primary surgery from 2003 to 2008 at our hospital were recruited. Patient QOL was evaluated using the University of Washington Quality of Life Questionnaire, version 4. Statistical analysis was conducted using a paired-samples t test and multiple stepwise linear regression with Statistical Package for Social Sciences, version 11.5 (SPSS, Chicago, IL). RESULTS: At 1 year after surgery, the appearance, activity, speech, swallowing, shoulder function, salivary, and taste domain scores were significantly lower than the preoperative scores (P < .05). However, the pain, anxiety, and mood scores were significantly better 1 year after surgery (P < .05). The overall QOL had increased greatly 1 year after surgery but did not reach the pretreatment level. Multiple stepwise linear regression analysis showed that the main factors affecting QOL were radiotherapy, advanced clinical stage (P < .05), socioeconomic status, and patient age. Radiotherapy, advanced clinical stage (P < .05), socioeconomic status, and age (P < .05) were independently associated with QOL. CONCLUSIONS: The patients with tongue cancer who have been diagnosed and treated early might have a better QOL. A greater socioeconomic status can also improve the QOL of patients with tongue cancer after primary surgery.

Inhibition of ameloblastoma invasion in vitro and in vivo by inhibitor of metalloproteinase-2 activity.

BACKGROUND: Ameloblastoma is an odontogenic benign tumor characterized by local invasiveness and most of its local recurrences clinically result from local invasion. This study used matrix metalloproteinase-2 (MMP-2) inhibitor I (MMP-2I) to investigate the role played by MMP-2 activity in the local invasiveness of ameloblastoma. METHODS: The cells and xenografts of ameloblastoma were treated with MMP-2I and treatment group were compared with the control group. In vitro, the invasive activity of tumor cells was assayed in transwell cell culture chamber. Gelatinolytic activity of gelatinases and MMP-2/tissue inhibitor of matrix metalloproteinase (TIMP-2) protein expression was detected using gelatin zymography and flow cytometry. The cell viability and adhesion were evaluated using methyl thiazol tetrazolium. In vivo, bilateral subrenal capsule xenograft transplantation of ameloblastoma was performed in 10 nude mice and the invasion of ameloblastoma into the renal parenchyma was observed. RESULTS: Active-MMP-2 of conditioned media was significantly lower in treatment group than in the control group. Accordingly, potential of in vitro cell invasion, adhesion and in vivo tumor invasion were also significantly lower in the treatment group than in the control group. CONCLUSIONS: Inhibitor of MMP-2 activity suppressed the local invasive capability of ameloblastoma by decreasing MMP-2 activity. MMP-2 activity is in relation with invasive capacity of ameloblastoma.

Differences between epithelial and mesenchymal human tongue cancer cell lines in experimental metastasis.

Distant metastasis represents the outcome with the worst prognosis for various types of malignant tumors, but little is known regarding the impact of interacting epithelial and mesenchymal phenotypic cancer cells within its etiopathogenesis. In a novel animal model, 48 male athymic Balb/c nude mice underwent subcutaneous and intravenous injection of human tongue cancer cell lines of green fluorescent mesenchymal and red fluorescent epithelial phenotypes, in order to visualize and monitor eventual phenotypic interaction in lung metastasis as well as experimental metastasis in in vivo, ex vivo and histopathological analyses. While the epithelial, but not the mesenchymal, phenotypic human tongue cancer cell line led to direct metastasis in the lungs when injected intravenously, neither of them, even when injected in combination, were able to establish distant metastasis. The results of the present study provide evidence regarding the role of epithelial phenotypic cancer cells in the release of experimental metastasis following tail vein injection in male athymic Balb/c nude mice, in addition to proving fluorescent human tongue cancer cells may be reliably detected under a fluorescence microscope even 8 weeks after the two injection types.

[Retracted] Long non­coding RNA MALAT1 interacts with miR­124 and modulates tongue cancer growth by targeting JAG1.

We wish to retract our research article entitled "Long non­coding RNA MALAT1 interacts with miR­124 and modulates tongue cancer growth by targeting JAG1" published in Oncology Reports 37 2087­2094, 2017. Following the publication of this article, it was drawn to our attention that this paper bore numerous similarites with an article published previously in the journal OncoTargets and Therapy. Although all the data reported in our study were original, we recognize that it was not appropriate that we should have modelled our paper on previously published articles as a template on which to base the writing of our paper. Therefore, we have agreed to follow the Editor's recommendation that this paper be retracted from the publication. All the named authors agree to this retraction. We sincerely apologize to the Editor and the readership of the Journal for our action, and regret any inconvenience this has caused. [the original article was published in the Oncology Reports 37: 2087­2094, 2017; DOI: 10.3892/or.2017.5445].

[Effect analysis of modified radical surgery in tongue cancer at early stages].

OBJECTIVE: To evaluate the oncologic efficacy, surgical safety, postoperative beauty, and morbidity of an ideal procedure of modified radical surgery in tongue cancer at early stages. METHODS: Six patients with early stage squamous cell carcinoma of tongue underwent a procedure of modified radical surgery, in which an inconspicuous incision was used, the external jugular vein, sternocleidomastoid muscle, spino-accessory nerve, and greater auricular nerve were preserved, half of glosso and mouth floor were excised, and the defect was repaired instantly. Recurrence of glosso, mouth floor and neck, regional edema, shoulder dysfunction, auricle sensibility, oral cavity function and beautiful outlook on the face and neck were evaluated clinically. RESULTS: Compared with classic radical neck dissection, this procedure of modified radical surgery showed an inconspicuous incision and pretty appearance, minor edema on face and neck, better shoulder function, and sensation of auricular skin. No recurred to the glosso, mouth floor, and neck was found during follow-up. The patients showed better oncological safety, pretty appearance of tongue, and better oral function of speech, swallow and mastication. CONCLUSION: This ideal procedure of modified radical surgery in tongue cancer at early stages lessens or avoids destruction of face and neck, shoulder malfunction, numbness of auricular skin, and oral dysfunction of speech, swallow and mastication without impairment on the oncologic safety of the radical surgery, and improves the quality of life of the patients.

Inhibition of periderm removal in all-trans retinoic acid-induced cleft palate in mice.

Cleft palate is a common craniofacial birth defect. The aim of the present study was to investigate the effect of excess all-trans retinoic acid (atRA) on periderm removal and the disappearance of basal medial edge epithelial (MEE) cells during palatogenesis, particularly during the stage prior to contact. atRA (200 mg/kg) was administered to C57BL/6N mice at embryonic day (E) 12.0 by gavage. Fetal palates were processed and analyzed by histology and electron microscopy. Single palate shelf peridermal cells were removed and cultured in the presence of atRA (3 µM) only or in the presence of or the caspase inhibitor, Z-VAD (100 µM) only, for 48 h. Once cultured, morphological changes were analyzed by histological staining and electron microscopy. A TUNEL assay was used to detect apoptotic neurons. Paired palatal shelves with periderm removal were cultured in the presence of atRA (3 µM) only or in the presence of Z-VAD (100 µM) only for 48 h and analyzed by hematoxylin and eosin staining. At E14.5, medial edge epithelium periderm was retained in the atRA-treated palates but had been shed prior to contact in the control groups. In addition, atRA was revealed to disrupt the cell cycle in the periderm by downregulating p21. Furthermore, atRA inhibited apoptosis in the periderm and basal MEE cells; however, atRA exhibited no effect on basement membrane degradation in single palatal organ culture. Additionally, once paired palates were cultured for 48 h, all of the groups in which the periderm had been removed exhibited confluence of the embryonic palatal mesenchyme. The present results suggest that periderm removal is inhibited in atRA-induced cleft palate in mice and that removal of the periderm contributes to EPM confluence in vitro.

Evidence of phenotypic stability after transduction of fluorescent proteins in two human tongue cancer cell lines.

OBJECTIVES: This study investigated the phenotypic stability and biological properties of two human tongue cancer cell lines after transduction of fluorescent proteins. DESIGN: The human tongue cancer cell lines UM1 and UM2 were cultured with GFP and RFP lentiviral particles stock for 72h. Cells with successful transduction of fluorescent proteins were selected in a medium containing G418 antibiotics for two weeks. The proliferation rates of parental and transduced cell lines were evaluated by their population doubling time (PDT). Transduction efficiency was assessed by fluorescence microscope and flow cytometry. The transduced cells in passage 1, 2, 10, 20 and 30 were collected to check the stability of fluorescent protein expression. Phenotypic stability of the transduced cells was detected by means of cell morphology, cell surface markers and cell function evaluating essay. RESULTS: The proliferation rates of the transduced cell lines showed no significant difference compared to their parental cells. Successful transduction with high efficiency (99% up) was demonstrated. High fluorescence expression on both transduced cells was detected until the thirtieth generation. UM1 and UM1-GFP displayed mesenchymal cell characteristics, while UM2 and UM2-RFP cell lines showed properties characteristic of epithelial. CONCLUSIONS: Two human tongue cancer cell lines of epithelial and mesenchymal phenotype respectively, have been successfully labelled with green and red fluorescent proteins. The fluorescence maintained a high expression rate over thirty generations without influencing the original morphological phenotype and cadherin expression.

Long non-coding RNA MALAT1 interacts with miR-124 and modulates tongue cancer growth by targeting JAG1.

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), a long non-coding RNA (lncRNA), was the earliest discovered to be correlated with cancer and contributes to the initiation and development of several types of tumors. Dysregulation of MALAT1 expression is frequently observed in many types of cancer such as gastric cancer, esophageal squamous cell carcinoma and glioma. To date, the role of MALAT1 and the underlying mechanisms in tongue cancer development remain unclear. In the present study, we studied the influence of MALAT1 on tongue cancer cell lines and clinical tongue cancer samples so as to detect its function and the underlying mechanism. In the present study, lncRNA-MALAT1 was specifically upregulated in tongue cancer cell lines and overexpression promoted tongue cancer cell growth by targeting miR-124. Knockdown of MALAT1 suppressed the growth and invasion of human tongue cancer cells and inhibited metastasis in vitro and in vivo. In addition, miR-124-dependent jagged1 (JAG1) regulation was required for MALAT1-induced tongue cancer cell growth. Our data revealed that MALAT1 inhibited tongue cancer cell growth and metastasis through miR-124-dependent JAG1 regulation. In conclusion, we revealed that MALAT1 may play an oncogenic role by increasing proliferation and metastasis of tongue cancer and is a potential therapeutic target in human tongue cancer.

[Combined repair of large defect caused by radical surgery of advanced tongue cancer with rib-major pectoralis myocutaneous flap carrying costal parietal pleura].

OBJECTIVE: To explore the clinical value and safety of using rib-major pectoralis myocutaneous flap carrying costal parietal pleura in combined repair of large soft and hard tissue defect caused by radical surgery of advanced tongue cancer. METHODS: Six patients with advanced tongue carcinoma involving the floor of mouth and mandible were performed combined radical neck dissection with glossectomy and mandibulectomy, which caused large soft and hard tissue defect. Six rib-major pectoralis myocutaneous flaps carrying costal parietal pleura were transferred for immediate repair of the large defects. The rib flaps were applied for the repair of mandible, and the major pectoralis myocutaneous flaps were applied for the reconstruction of tongue and floor of mouth. RESULTS: Six patients recovered well after operation. Six rib-major pectoralis myocutaneous flaps carrying costal parietal pleura survived well; the wounds of surgical incision of the oral cavity, neck, and chest healed up. The reconstructed tongue and the lower face appearance were satisfactory, the occlusion relationships were normal; the speaking as well as swallowing functions recovered. CONCLUSIONS: It's safe and reliable to use rib-major pectoralis myocutaneous flap carrying costal parietal pleura to repair large soft and hard tissue defect in oral and maxillofacial region. Opening pleural cavity and harvest costal parietal pleura would not influence patients' thoracic movement and breath function and would not cause other complications. It's simple and safe for harvesting the composite flap. Carrying costal parietal pleura assures the sufficient blood supply of rib in the composite flap.

Tumorigenicity and Validity of Fluorescence Labelled Mesenchymal and Epithelial Human Oral Cancer Cell Lines in Nude Mice.

Tumorigenicity and metastatic activity can be visually monitored in cancer cells that were labelled with stable fluorescence. The aim was to establish and validate local and distant spread of subcutaneously previously injected fluorescence transduced human tongue cancer cell lines of epithelial and mesenchymal phenotype in nude mice. A total of 32 four-week-old male athymic Balb/c nude mice were randomly allocated into 4 groups (n = 8). A single dose of 0.3 mL PBS containing 1 × 107 of four different cancer cell-lines (UM1, UM1-GFP, UM2, and UM2-RFP) was injected subcutaneously into the right side of their posterolateral back. Validity assessment of the labelled cancer cells' tumorigenicity was assessed by physical examination, imaging, and histology four weeks after the injection. The tumor take rate of cancer cells was similar in animals injected with either parental or transduced cancer cells. Transduced cancer cells in mice were easily detectable in vivo and after cryosection using fluorescent imaging. UM1 cells showed increased tumor take rate and mean tumor volume, presenting with disorganized histopathological patterns. Fluorescence labelled epithelial and mesenchymal human tongue cancer cell lines do not change in tumorigenicity or cell phenotype after injection in vivo.