Mutual regulation between miR-21 and the TGFß/Smad signaling pathway in human bronchial fibroblasts promotes airway remodeling.

OBJECTIVE: Airway remodeling is an important pathological feature of asthma. Excessive deposition of extracellular matrix (e.g., collagen) secreted from fibroblasts is a major factor contributing to airway remodeling. Currently, the mechanism by which collagen continues to be oversynthesized in the airway remains unclear. In this study, we investigated the role of the microRNA-21 (miR-21) and TGFß/Smad signaling pathway in human bronchial fibroblasts (HBFs), and explored the regulatory mechanism of airway remodeling. METHODS: HBFs were cultured in vitro and treated with the transforming growth factor ß (TGFß), receptor inhibitor (SB431542), and TGFß1. miR-21 and Smad7 overexpressing lentiviruses, as well as an miR-21 interfering lentivirus were constructed and transfected into HBFs. Western blotting was used to determine the expression of airway remodeling-related proteins and proteins in the TGFß/Smad signaling pathway. miR-21 expression was measured by quantitative real-time PCR. RESULTS: The high expression of miR-21 induced by TGFß1 was reduced following the treatment with the SB431542 in HBFs. Smad7 overexpression inhibited the elevated expression of the COL I protein induced by miR-21 overexpression in HBFs. Inhibiting miR-21 expression upregulated the level of Smad7 protein, thus reducing the expression of airway remodeling-related proteins induced by TGFß1 stimulation in HBFs. CONCLUSIONS: TGFß1 can induce miR-21 expression in HBFs through the TGFß/Smad signaling pathway to promote airway remodeling. miR-21 downregulates Smad7, activates the TGFß/Smad signaling pathway, and promotes airway remodeling. Mutual regulation between miR-21 and the TGFß/Smad signaling pathway in HBFs promotes airway remodeling.

The lncRNA CRNDE promotes colorectal cancer cell proliferation and chemoresistance via miR-181a-5p-mediated regulation of Wnt/ß-catenin signaling.

BACKGROUND: With more than 600,000 mortalities each year, colorectal cancer (CRC) is the third most commonly diagnosed type of cancer worldwide. Recently, mechanisms involving noncoding RNAs have been implicated in the development of CRC. METHODS: We examined expression levels of lncRNA CRNDE and miR-181a-5p in 64 cases of CRC tissues and cell lines by qRT-PCR. Gain-of-function and loss-of-function assays were performed to examine the effect of CRNDE and miR-181a-5p on proliferation and chemoresistance of CRC cells. Using fluorescence reporter and western blot assays, we also explored the possible mechanisms of CRNDE in CRC cells. RESULTS: In this study, we found that the expression levels of the CRNDE were upregulated in CRC clinical tissue samples. We identified microRNA miR-181a-5p as an inhibitory target of CRNDE. Both CRNDE knockdown and miR-181a-5p overexpression in CRC cell lines led to inhibited cell proliferation and reduced chemoresistance. We also determined that ß-catenin and TCF4 were inhibitory targets of miR-181a-5p, and that Wnt/ß-catenin signaling was inhibited by both CRNDE knockdown and miR-181a-5p overexpression. Significantly, we found that the repression of cell proliferation, the reduction of chemoresistance, and the inhibition of Wnt/ß-catenin signaling induced by CRNDE knockdown would require the increased expression of miR-181a-5p. CONCLUSIONS: Our study demonstrated that the lncRNA CRNDE could regulate the progression and chemoresistance of CRC via modulating the expression levels of miR-181a-5p and the activity of Wnt/ß-catenin signaling.

[Role of serum neutrophil elastase determination in the diagnosis of acute exacerbation of asthma in preschool children].

OBJECTIVE: To study the role of serum neutrophil elastase (NE) level in acute exacerbation of asthma in preschool children. METHODS: A total of 85 preschool children who were diagnosed with asthma between January 2008 and January 2010 were classified into acute exacerbation group (n=44) and non-acute exacerbation group (n=41). Thirty-five children who received physical examination served as the control group. The enzyme-linked immunosorbent assay was used to determine the serum levels of NE and interleukin-8 (IL-8). The receiver operating characteristic (ROC) curve was used for NE evaluation. RESULTS: Both the acute and non-acute exacerbation groups had higher serum levels of NE and IL-8 than the control group, and the acute exacerbation group had significantly higher serum levels of NE and IL-8 than the non-acute exacerbation group (P<0.05). The serum level of NE was positively correlated with that of IL-8 (r=0.48, P<0.05). With serum NE level >27.73 µg/L as the cut-off value for diagnosing acute exacerbation of asthma, the sensitivity was 65.9%, the specificity was 95.1%, and the area under the ROC curve was 0.87 (P<0.01). CONCLUSIONS: The determination of serum NE level in preschool children with asthma helps to diagnose the acute exacerbation of asthma.

[Changes in serum inflammatory factors in wheezing infants with community-acquired pneumonia].

OBJECTIVE: To study whether infantile wheezing pneumonia has similar immune mechanisms to asthma by determining the levels of serum inflammatory factors in wheezing infants with community-acquired pneumonia (CAP). METHODS: Forty-two infants with CAP but without wheezing, 47 infants with CAP and wheezing, and 30 healthy infants as a control were recruited in the study. The peripheral blood levels of C-reactive protein, procalcitonin, soluble triggering receptor expressed on myeloid cell-l, interferon-γ, interleukin-4, interleukin-10, and periostin were compared in the three groups. RESULTS: The serum levels of procalcitonin, soluble triggering receptor expressed on myeloid cell-l, interleukin-4 and interleukin-10 in the two CAP groups were higher than in the control group (P<0.05). The ratio of interferon-γ/interleukin-4 in the wheezing pneumonia group was lower than in the non-wheezing pneumonia and control groups (P<0.05). The serum level of periostin in the wheezing pneumonia group was higher than in the non-wheezing pneumonia and control groups (P<0.05). CONCLUSIONS: The unbalanced ratio of interferon-γ/interleukin-4 and airway eosinophilic inflammation in wheezing infants with pneumonia suggest infantile pneumonia with wheezing may has similar immune mechanisms to asthma.

Circular RNA Fibroblast Growth Factor Receptor 1 Promotes Pancreatic Cancer Progression by Targeting MicroRNA-532-3p/PIK3CB Axis.

OBJECTIVE: The aim of the study is to explore the contribution and mechanism of circular RNA fibroblast growth factor receptor 1 (circFGFR1) in pancreatic ductal adenocarcinoma (PDAC) progression. METHODS: Expressions of circFGFR1, microRNA (miR)-532-3p, and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta (PIK3CB) were assessed by quantitative real-time polymerase chain reaction or in situ hybridization. Fluorescence in situ hybridization determined the subcellular localization of circFGFR1. Immunohistochemistry was used to detect PIK3CB expression in PDAC tissues. Cell growth was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and colony formation assays. Wound healing, transwell, and flow cytometry assays examined the migration, invasion, and apoptosis. Dual-luciferase and RNA pull-down assay verified the interactions between circFGFR1/PIK3CB and miR-532-3p. In vivo xenograft tumor growth and lung metastasis were assessed in nude mice. RESULTS: Functionally, knockdown of circFGFR1 restrained in vitro PDAC cell growth, migration, invasion, and in vivo xenograft tumor growth and lung metastasis. In addition, circFGFR1 could sponge miR-532-3p to upregulate PIK3CB level. Rescue experiments revealed that the tumor-suppressive effects caused by miR-532-3p mimics could be reversed by circFGFR1 or PIK3CB overexpression. CONCLUSIONS: Our data revealed that circFGFR1 driven the malignant progression of PDAC by targeting miR-532-3p/PIK3CB axis, suggesting that inhibition of circFGFR1 might be considered as a therapeutic target for PDAC.

[Roles of allergen testing in vitro and impulse oscillometry for lung function measurements in children with cough variant asthma].

OBJECTIVE: To study the roles of allergen testing in vitro and impulse oscillometry for lung function measurements in preschool children with cough variant asthma (CVA). METHODS: ethodsForty-four preschool children with acute asthma, 41 with chronic asthma, 46 with CVA, and 35 healthy preschool children as control were recruited in the study. Inhaled allergen, food allergen, and mite-specific IgE were determined by Pharmacia UniCAP System. Serum eosinophil cationic protein (ECP) and total IgE levels were measured. Lung function was assessed by impulse oscillometry. RESULTS: The positive rates of inhaled allergen and food allergen, and total IgE levels in the CVA, acute asthma and chronic asthma groups were higher than those in the control group (P<0.01). However, no significant differences were found among the three case groups. The serum ECP levels in the CVA group were lower than those in the acute asthma group (P<0.01), but did not show differences when compared with the chronic asthma group. The impulse oscillometry demonstrated that the respiratory total impedance (Zrs), airway resistance at 5 Hz (R5), airway resistance at 20 Hz (R20), subtracting R5 from R20 (R5-R20) and resonant frequency (Fres) in the CVA, acute asthma and chronic asthma groups were higher than those in the control group (P<0.01). Zrs, R5, R20, R5-R20, and Fres in the CVA and chronic asthma groups were lower than those in the acute asthma group (P<0.01). Serum ECP levels were positively correlated with Zrs, R5, R5-R20 and Fres (P<0.05) in the CVA and chronic asthma groups. CONCLUSIONS: The measurements of allergens, serum ECP and impulse oscillometry for lung function are helpful for the evaluation of airway inflammation and airway obstruction in preschool children with CVA.

Micropatterned Fibrous Scaffold Produced by Using Template-Assisted Electrospinning Technique for Wound Healing Application.

This paper describes the fabrication of a structural scaffold consisting of both randomly oriented nanofibers and triangular prism patterns on the scaffold surface using a combination technique of electrospinning and collector templates. The polycaprolactone (PCL) nanofibers were electrospun over a triangular prism pattern mold, which acted as a template. The deposited scaffold was removed from the template to produce a standalone structural scaffold of three-dimensional micropatterned nanofibers. The fabricated structural scaffold was compared with flat randomly oriented nanofibers based on in vitro and in vivo studies. The in vitro study indicated that the structural scaffold demonstrated higher fibroblast cell proliferation, cell elongation with a 13.48 ± 2.73 aspect ratio and 70% fibroblast cell orientation compared with flat random nanofibers. Among the treatment groups, the structural scaffold escalated the wound closure to 92.17% on day 14. Histological staining of the healed wound area demonstrated that the structural scaffold exhibited advanced epithelization of the epidermal layer accompanied by mild inflammation. The proliferated fibroblast cells and collagen fibers in the structural scaffold appeared denser and arranged more horizontally. These results determined the potential of micropatterned scaffolds for stimulating cell behavior and their application for wound healing.

Controlling cell elongation and orientation by using microstructural nanofibre scaffolds for accelerating tissue regeneration.

The topographic surface conditions of scaffolds can regulate cellular behaviours, such as by stimulating cellular migration and morphological changes to wound sites and have the potential to promote tissue regeneration. In this research, four types of engineered topographic surfaces, including arrays of hemisphere, pyramid, semi-cylinder, and triangle prism microstructures, were patterned on silicon moulds using microfabrication processes. The microstructural patterns were transferred onto the surface of polycaprolactone membranes and nanofibrous scaffolds by combining with the moulding approach and electrospinning technique, respectively. In vitro experimental results demonstrated that the triangular microstructural nanofibre provided a strong guiding performance to the filopodia of cultured C2C12 myoblast cells, thus inducing cellular elongation and alignment in the longitudinal direction and forming an elongated cell morphology. The cultured cells rapidly transitioned into an elongated morphology at an aspect ratio of 17.33 after 24 h of incubation, with 70% of the cell elongates aligning with the direction of triangular microstructural patterns. The cells cultured on the triangular microstructural nanofibre elongated four-fold compared with those in the flat nanofibre scaffold. Moreover, an in vivo study showed that wounds treated with the triangular microstructural nanofibre scaffold achieved 95.04% wound closure after 14 days and completed the reepithelialisation with an ordered collagen arrangement. Therefore, we believe that the engineered triangular nanofibrous scaffold may accelerate tissue regeneration and has potential for wound healing applications.

Development of Thermo-Responsive Polycaprolactone-Polydimethylsiloxane Shrinkable Nanofibre Mesh.

A thermally activated shape memory polymer based on the mixture of polycaprolactone (PCL) and polydimethylsiloxane (PDMS) was fabricated into the nanofibre mesh using the electrospinning process. The added percentages of the PDMS segment in the PCL-based polymer influenced the mechanical properties. Polycaprolactone serves as a switching segment to adjust the melting temperature of the shape memory electro-spun PCL-PDMS scaffolds to our body temperature at around 37 °C. Three electro-spun PCL-PDMS copolymer nanofibre samples, including PCL6-PDMS4, PCL7-PDMS3 and PCL8-PDMS2, were characterised to study the thermal and mechanical properties along with the shape memory responses. The results from the experiment showed that the PCL switching segment ratio determines the crystallinity of the copolymer nanofibres, where a higher PCL ratio results in a higher degree of crystallinity. In contrast, the results showed that the mechanical properties of the copolymer samples decreased with the PCL composition ratio. After five thermomechanical cycles, the fabricated copolymer nanofibres exhibited excellent shape memory properties with 98% shape fixity and above 100% recovery ratio. Moreover, biological experiments were applied to evaluate the biocompatibility of the fabricated PCL-PDMS nanofibre mesh. Owing to the thermally activated shape memory performance, the electro-spun PCL-PDMS fibrous mesh has a high potential for biomedical applications such as medical shrinkable tubing and wire.

[Exploration on the modification of macroscopic classification of colorectal cancer].

OBJECTIVE: To explore the feasibility and clinical significance of a modified macroscopic classification of colorectal cancer. METHODS: The data of 1379 patients with colorectal cancer surgically treated between 1975 and 2003 were retrospectively analyzed. The patients were divided into four groups according to the primary macroscopic appearance: protruding type (group 1), local ulcerative type (group 2), invasive type (group 3) and non-invasive ulcerative type (group 4). The new classification system was simplified into two types: non-invasive type (group A, including group 1 and 2) and invasive type (group B, including group 3 and 4). The histo-differentiation, invasive depth into the intestinal wall, distance and number of lymph node metastasis and 5-year survival rate were analyzed and compared among the groups. RESULTS: There was no significant difference between group 1 and 2, and between group 3 and 4 in histodifferentiation, invasive depth into the intestinal wall, distance and number of lymph node metastasis and 5-year survival rate (P>0.05). However, after modification of the primary macroscopic classification, a significant difference was observed in all the above mentioned parameters between group A and group B (P<0.05). CONCLUSION: Our results demonstrate that the clinicopathological characteristics of the group 1 and 2, and of the group 3 and 4 are similar to each other. So it is reasonable to merge the protruding type and local ulcerative type into non-invasive type, while invasive type and non-invasive ulcerative type into invasive type. This simplified macroscopic classification should be practical and instructive in diagnosis, treatment and prognosis of colorectal cancer.

[Study on the determination of molybdenum and other elements in ferromolybdenum alloy by EDX].

Some kinds of the high impurity ferromolybdenum alloy occurred in the ferroalloy market recently. The analysis result of the element Mo in high impurity ferromolybdenum alloy is not accurate using the routine chemical method, because of the coprecipitation of the impurity elements, compared with the final Mo content of poured steel ingots. The energy dispersive X-ray spectroscopy (EDX) was applied to the analysis of ferromolybdenum alloy. The results show that EDX is an appropriate way to the determination of element Mo and Si in ferromolybdenum alloy, but can not be used to analyze element P, S and Cu. As the morphology of the sample has a strong influence on the analysis results, scanning electron microscope (SEM) was used to observe the morphology of the sample. The detection precision of the element shall be raised, when the powder sample of the ferromolybdenum alloy are pressed into thinner disc sample using hydraulic universal testing machine.

Detection of active fraction of glycogen synthase kinase 3beta in cancer cells by nonradioisotopic in vitro kinase assay.

Glycogen synthase kinase 3beta (GSK3beta) is a well-known marker and potential therapeutic target in non-insulin-dependent diabetes mellitus and Alzheimer's disease. Our recent demonstration that GSK3beta has a previously unrecognized role in colorectal cancer facilitates the development of a nonradioisotopic in vitro kinase assay (NRIKA) for detecting GSK3beta activity in gastrointestinal cancer cells. The NRIKA uses a sequential combination of immunoprecipitations to isolate GSK3beta in sample cells' lysates, and an in vitro kinase reaction that uses recombinant beta-catenin protein (substrate) and nonradioisotopic ATP, followed by immunoblotting to detect beta-catenin phosphorylated in serine 33, 37 and/or threonine 41 residues. The NRIKA detected higher expression of active GSK3beta in stomach, colon, pancreas and liver cancer cell lines than in human embryonic kidney cells (HEK293) considered nonneoplastic. Inhibition of cancer cell-derived GSK3beta activity by GSK3beta inhibitors (SB-216763, AR-A014418) was detected by the NRIKA. GSK3beta inhibition attenuated survival and proliferation and induced apoptosis in all types of cancer cells but not in HEK293. These findings supported the idea that the pathologic roles of GSK3beta are definite and common in various types of cancer. The NRIKA provides a basis for evolving a high-throughput tool for testing substances for GSK3beta inhibition, and for screening and identifying novel GSK3beta inhibitors with a view to discovering drugs for treatment of cancer as well as non-insulin-dependent diabetes mellitus and Alzheimer's disease.

[Effect of bacillus calmette-guerin treatment on airway inflammation and T regulatory cells in mice with asthma].

OBJECTIVE: Previous studies have shown that bacillus calmette-guerin (BCG) can deviate TH2 response toward TH1 response, resulting in a suppressive effect on the development of asthma/atopy. This study examined the effect of BCG treatment on regulatory T cells in asthmatic mice to investigate the possible mechanism. METHODS: Kunming mice were sensitized and challenged with ovalbumin (OVA) to establish asthmatic models. Asthmatic mice were injected intradermally with BCG five days before and after sensitization. After 24 hrs of last challenge, bronchoaveolar lavage fluid (BALF) and peripheral blood were collected . The total cells and eosinophils were counted in the BALF. The percentage of CD4(+) CD25(+) in peripheral blood was detected with flow cytometry. Single spleen cell suspension was prepared and cultured in 1640 medium for 48 hrs and then the cytokine IL-10 level in the supernatant was determined using ELISA. The mice which were challenged with normal saline were used as the Normal control group. RESULTS: The number of total cells and eosinophils in BALF in asthmatic mice [(27.27 +/- 5.36) x 10(7)/L and (6.59 +/- 1.32) x 10(7)/L respectively] were more than in the Normal control group [(1.52 +/- 0.36) x 10(7)/L and zero respectively] (P < 0.01). The number of total cells and eosinophils in BALF in asthmatic mice were reduced after BCG treatment [(13.71 +/- 3.17) x 10(7)/L and (1.43 +/- 0.37) x 10(7)/L respectively] (P < 0.01). The percentage of CD4(+) CD25(+) in peripheral blood of asthmatic mice [(11.59 +/- 1.33)%] was noticeably lower than that of the Control group [(13.66 +/- 1.68)%] (P < 0.01), but increased significantly in asthmatic mice after BCG treatment [(14.40 +/- 2.70)%] (P < 0.05). The IL-10 level in spleen cell supernatant in the BCG-treated group (7.79 +/- 1.34 pg/mL) also increased compared with that in the untreated asthmatic mice (5.54 +/- 0.66 pg/mL) (P < 0.01). CONCLUSIONS: BCG can markedly inhibit the airway inflammation in asthmatic mice possibly by promoting the production of regulatory T cells.

[Expression of matrix metalloproteinase 2 in oral verruvous carcinoma and squamous cell carcinoma].

OBJECTIVE: To determine the expression of MMP2 mRNA in oral verruvous carcinoma and squamous cell carcinoma. METHODS: Thirty cases were divided into 3 groups: verruvous carcinoma (n = 10), well-differentiated squamous cell carcinoma (n = 15) and moderately or poorly differentiated squamous cell carcinoma (n = 5). Reverse transcription polymerase chain reaction (RT-PCR) was used to test the expression of MMP2 mRNA in the carcinoma tissues and matched normal tissues from 3 groups above. RESULTS: The expression of MMP2 mRNA in the carcinoma tissues was significantly higher than that in their matched normal tissues (P < 0.05). The expression of MMP2 mRNA in verruvous carcinoma was significantly higher than that in well-differentiated and moderately or poorly differentiated squamous cell carcinoma (P < 0.05). However, the expression of MMP2 mRNA was not obviously different between well-differentiated and moderately or poorly differentiated squamous cell carcinoma (P > 0.05). CONCLUSION: The expression of MMP2 mRNA in oral verruvous carcinoma and squamous cell carcinoma tissues was significantly higher than that in their matched normal tissues. The expression of MMP2 mRNA in verruvous carcinoma was significantly higher than that in squamous cell carcinoma.

Effect of extended radical resection for rectal cancer.

AIM: To discuss the rationality of extended radical resection (ERR) and to guide the surgical treatment of rectal cancer. METHODS: Total 211 patients who underwent ERR from 1981 to 1987 (follow-up rate of 94.8 %) were selected to study the patterns of lymphatic metastasis and therapeutic effect. The control group was made of 293 patients with rectal cancer who underwent conventional radical resection (CRR) and its follow-up rate was 98.5 %. The lymph node specimens, obtained by the triple-approach lymph node resection during the radical resection of rectal cancer, were studied by conventional pathological method. The extended radical resection, guided by the patterns of lymphatic metastasis, was applied in the clinical practice. RESULTS: The incidence of lymphatic metastasis in Chinese patients with advanced rectal cancer was 43.6 %, and that of the upper 2nd and 3rd groups and the lateral group was 14.2 %, 10.9 % and 11 % respectively. The 5,10-year-survival rates of the ERR were 68.0 % and 47.0 %, respectively, which were much higher than those of the conventional radical resection (42.9 % and 25.3 %). CONCLUSION: The ERR for rectal cancer removes all the lymph nodes, prevents possible metastasis and finally improves the survival rate.

Reversal of 5-flouroucial resistance by adenovirus-mediated transfer of wild-type p53 gene in multidrug-resistant human colon carcinoma LoVo/5-FU cells.

AIM: To observe the reversal effects of wide-type p53 gene on multi-drug resistance to 5-FU (LOVO/5-FU). METHODS: After treatment with Ad-p53, LOVO/5-FU sensitivity to 5-Fu was investigated using tetrazolium dye assay. Multidrug resistance gene-1 (MDR1) gene expression was assayed by semi-quantitative reverse transcription-polymerase chain reaction and the expression of p53 protein was examined by Western blotting. RESULTS: The reversal activity after treatment with wide-type p53 gene was increased up to 4.982 fold at 48 h. The expression of MDR1 gene decreased significantly after treatment with wide-type p53 gene, and the expression of p53 protein lasted for about 5 d, with a peak at 48 h, and began to decrease at 72 h. CONCLUSION: Wide-type p53 gene has a remarkable reversal activity for the high expression of MDR1 gene in colorectal cancers. The reversal effects seem to be in a time dependent manner. It might have good prospects in clinical application.

[On rationality of total mesorectal excision for rectal cancer].

OBJECTIVE: To discuss the rationality and clinical significance of total mesorectal excision (TME) assessed by the pattern of local recurrence of rectal cancer. METHODS: 207 rectal cancer patients treated by surgery from 1975 to 2001 who developed local recurrence were retrospectively studied as to the option of treatment based on their outcome. 172 of these 207 patients had complete record of follow-up with a follow-up rate of 83.0%. RESULTS: The local recurrent sites of 207 patients were: anastomotic stoma 71, mesorectum 65, perineum 50, lymph nodes 59, multiple recurrences 89 and others sites rare. Among 81 patients who had been first treated by anterior resection (AR), 58 were again treated by abdomino-perineal resection (APR). Among 102 patients who had been treated by APR, 38 were treated by perineal tumor resection, 15 were treated by total and posterior pelvic organ resection, and 18 were supplemented with lymph node resection. By all modalities, the resection rate was 66.1% (137/207), in which the curable resection rate was 46% (63/137). The 5-year survival rate of patients with tumor resection was 23.3% (32/137), those treated by curable resection was 34.9% (22/63). CONCLUSION: From the pattern of local recurrence, the operation for middle-lower rectal cancer must follow the principle of total mesorectum excision (TME). Patients with local recurrence should be energetically treated in the second operation if tolerable.

Site-specific lymphatic mapping relative to lingual septum in localizing the regional lymph nodes of tongue - an animal study.

BACKGROUND AND OBJECTIVES: With technical adaptations, recent studies showed SLNB could predict cervical nodes status of head and neck carcinoma with high accuracy. However, as for tongue carcinoma, such technical adaptations seem to be not enough because the tongue has peculiar characteristic which may demand a specific procedure for accurate lymphatic mapping. This investigation explored the effect of lingual septum on lymphatic mapping of tongue to provide data for achieving an accurate lymphatic mapping for managing early tongue carcinoma. METHODS: Four doses of Methylene Blue were injected into various parts of 64 rabbits' tongue, then diffusion range of Methylene Blue in tongue and sites of cervical nodes stained blue were noted. Finally, the tongues were resected for further histological examination and morphometric assessments. RESULTS: There was lingual septum in the tongue and the diffusing of Methylene Blue could be terminated by lingual septum. Blue-stained nodes were identified in 84 lateral necks of 60 rabbits. CONCLUSIONS: A site-specific way of lymphatic mapping relative to lingual septum should be developed for staging early tongue carcinoma.

MicroRNAs contribute to the chemoresistance of cisplatin in tongue squamous cell carcinoma lines.

MicroRNAs (miRNAs) are small non-coding RNAs that function as negative regulators of gene expression. They are strongly implicated in human cancers, including oral squamous cell carcinoma (OSCC). Evidence for the involvement of miRNAs as important regulators of chemosensitivity and chemoresistance in OSCC is not well understood. In this study, miRNA microarray was firstly used to compare the differential miRNAs levels between the cisplatin-sensitive tongue squamous cell carcinoma line (Tca8113) and its cisplatin-resistant subline (Tca/cisplatin). Three miRNAs of miR-21, -214, and -23a were validated by miRNAs real-time PCR, and intervened by anti-miRNA oligonucleotides (miR-214 and -23a) and pre-miRNA plasmid transfection (miR-21). Further relationship between miR-23a and DNA topoisomerase II beta (TOP2B) on the chemoresistance against cisplatin was studied. There were 19 out of 480 differential miRNAs between the Tca8113 and Tca/cisplatin cells. miR-214 and -23a were found increased as with chemoresistance against cisplatin in the Tca/cisplatin cells while miR-21 was found decreased as with chemosensitivity for cisplatin in the Tca/cisplatin cells. Intervention of these three miRNAs could decrease the chemoresistance against cisplatin in Tca/cisplatin cells. Transfection of anti-miR-23a into the Tca/cisplatin cells could increase the TOP2B protein expression. Our results suggest the existence of differential miRNAs with chemosensitivity and chemoresistance between the cisplatin-sensitive and resistant tongue squamous cell carcinoma lines. miR-21 serves as a chemosensitive miRNA, while miR-214 and -23a serve as chemoresistant miRNAs in the tongue squamous cell carcinoma lines. miR-23a is an up-stream regulator of TOP2B to realize the chemoresistance of cisplatin.