[Correlative study between X-ray type after healing of congenital pseudarthrosis of the tibia in children and postoperative refracture].

Objective: To investigate the relationship between postoperative X-ray type in 2 years after healing of congenital pseudarthrosis of the tibia(CPT) and refracture of CPT in children. Methods: A retrospective study was performed on 67 children patients with Crawford type Ⅳ CPT who were treated with combined surgeries from December 2007 to August 2012.There were 46 male and 21 female patients with 37 cases with left CPT and 30 cases with right CPT. There were 12 cases with proximal tibia dysplasia, 56 cases with neurofibromatosis type 1. The median age when operation was 2.8 years(from 0.6 to 11.2 years). The patients were divided into three groups, CPT with hypertrophic group, CPT with mediate group and CPT with atrophic group, on the basis of ratio of healing cross-sectional area and transition zone in pseudarthrosis.The incidence of refracture in the three groups were investigated. Results: The refracture rates of three groups were 13%(5/38), 14%(3/21), 5/8, respectively.The refracture rate difference between CPT with hypertrophic group and CPT with mediate group was not statistically significant(P=0.590). The refracture rate of CPT with atrophic group was statistically significant lower than that of CPT with hypertrophic group and CPT with mediate group(P=0.007, 0.019). In addition, the refracture-free cumulative survival rate of CPT with hypertrophic group or CPT with mediate group was higher than that of CPT with atrophic group with the statistically significant difference(both P<0.05). And the refracture-free cumulative survival rate in CPT with hypertrophic group was lower than that in CPT with mediate group, the difference was not significant(P>0.05). Conclusion: After the union of CPT, patients with hypertrophic, mediate type X ray characteristic showed lower incidence of refracture than those with atrophic type.

The biological behaviour of human adenoid cystic carcinoma cells transduced with interleukin-2-gene.

Adenoid cystic carcinoma (ACC) of the salivary glands is a highly infiltrative malignant tumour with a tendency for lung metastasis. Gene therapy could be a potentially effective therapy for ACC and its metastasis. The aims of the study were: To transduce interleukin-2 (IL-2) gene into an ACC cell line with predisposition for lung metastasis (ACC-M); to compare the bioactivity of the gene-transduced cells and the parent cell line in vitro and in vivo. The IL-2 gene was transduced via a bicistronic retroviral vector into the ACC-M cells. The growth rate and DNA cell cycles of the parent ACC-M, the control viral vector AmGCEN, and the gene transduced AmIL-2 cell cultures were compared quantitatively and by flow cytometry, respectively. The tumourigenic ability of the three cell lines was verified by inoculation in athymic nude mice. The tumours developed were extracted and compared quantitatively and histologically. There was no difference in the growth rate and the DNA count between the ACC-M, AmGCEN, and AmIL-2 cell cultures. In the animal experiment, both the ACC-M and AmGCEN cells stimulated lung metastasis in all the mice, whereas there was no tumour found in the 1 x 10(6) AmIL-2 cells inoculation. On 3 x 10(6) AmIL-2 cells stimulation, three out of six mice developed tumours but the mass and volume of the tumours were smaller than the other two groups. Under light microscopy, the ACC-M tumours were mainly poorly differentiated with minimal cellular matrix, whereas the AmIL-2 tumours were well differentiated with ample matrix. The transduction of IL-2 gene can reduce the tumourigenicity of ACC-M cells and induces tumour cell differentiation in mice. The IL-2 gene can be a potential effective gene for the treatment of adenoid cystic carcinoma of salivary glands and its lung metastasis.

[Effects of millimeter wave combined with gamma-ray radiation on human tongue squamous cell carcinoma cell].

OBJECTIVE: To observe the effects of millimeter wave combined with (60)Co gamma-ray radiation on human tongue squamous cell carcinoma cell(Tca8113). METHODS: Using mm-wave combined with (60)Co gamma-ray to irradiate Tca8113 cell suspensions. The colony forming inhibition efficiency was observed three weeks later. 24 hours after radiation, the samples were observed under electron microscope. RESULTS: The study showed that the radiation could result in significant decrease of the colony forming efficiency (P<0.001). The inhibition efficiency was higher when the samples were exposed to high power density mm-wave or for a long duration (P<0.05 or P<0.01). The inhibiting effect in combined groups was more obvious than in singly radiated group (P<0.001 or P<0.05). There was no difference between 'R+HL' group and 'HL+R' group (P>0.05). Electron microscopy showed the cells' suprastructures had some injuries and regressive changes. And more serious changes existed in 'H' group. CONCLUSION: It could be concluded that mm-wave radiation could efficiently inhibit the colony forming capability of Tca8113 cell. And mm-wave radiation could lead to morphological changes of exterior or interior ultrastructures of Tca8113 cell.

[Experimental study of the effect of TNP-470 on human salivary adenoid cystic carcinoma cell in vitro].

OBJECTIVE: To investigate the effects of TNP-470 on human salivary adenoid cystic carcinoma cell line ACC M in vitro. METHODS: Cell proliferation was assessed by MTT assays and dye exclusion counting. Morphological changes of apoptosis were observed with fluorescent microscope. DNA ladder, apoptosis rate and cell cycle were examined by DNA agarose gel electronphores and fluorescence flow cytometry (FCM), respectively. RESULTS: The 50% inhibitory concentrations (IC50) of TNP-470 on ACC-M cells proliferation by MTT assays and dye exclusion counting were 40.68microg/ml and 46.38microg/ml. Apoptosis were observed by fluorescent microscope. DNA electrophoresis for the cells treated with TNP-470 showed brighter DNA ladder; Sub-G1 peak and G2/M arrest were also determined by FCM (P<0.01). CONCLUSION: TNP-470 has the effect of inducing apoptosis in ACC-M cells in vitro, which may be one of its antitumor mechanisms.

[Establishment and biological characteristics of cisplatin resistant cell line from human tongue squamous cell carcinoma Tca8113].

OBJECTIVE: To establish a CDDP-resistant cell line from human tongue squamous cell carcinoma cell line Tca8113 and to study the characteristics of this cell line. METHODS: The concentration of CDDP added to Tca8113 cells was increased gradually and the chemotherapeutic drug was added continually, which was to induce the CDDP resistance in Tca8113 cells. The doubling time of Tca8113/CDDP cell line and the oncogenicity of this cell line in nude mice indicated its change on CDDP resistance. The sensibility to drugs of the cell line was assayed by MTT method. DNA fragmentation was detected by electrophoresis. Immunocytochemistry was utilized to examine the expression of P-gp and GST-pi. RESULTS: Tca8113 /CDDP was established and the cells grew steadily in 10 micromol/L CDDP medium. The Tca8113/CDDP cells were also resistant to Vm-26, BLM, VDS, E-ADM, Taxol and other drugs in different degrees. The doubling time of the cells was 29.9 hours and after a month it changed to 16.7 hours. When inoculated in nude mice, the cells showed the typical features of squamous cell carcinoma ultra-structurally and pathologically. Agarose gel electrophoresis of DNA from Tca8113 cells treated with CDDP revealed "ladder"pattern, which was the symbol of apoptosis, but there was no "ladder" in Tca8113/CDDP. The expression of P-gp and GST-pi in Tca8113/CDDP was much higher than that in Tca8113, which was the proof of the up-regulating translation of MDR-1 mRNA. CONCLUSION: CDDP induced Tca8113 cells resistance, and resistant cell line Tca8113/CDDP was established. The multi-drug resistance of Tca8113/CDDP cells were related to the GSH/GST system.

[The expression of noncollagenous proteins and the tumor differentiation in oral maxillofacial osteosarcoma].

OBJECTIVE: To study the significance of the osteocalcin bone sialoprotein and osteopontin in cell differentiation and matrix mineralization of osteosarcoma. METHODS: Use immunohistochemistry method to detect the osteocalcin bone sialoprotein and osteopontin expressions in tumor cells and matrix of 50 cases of osteosarcoma. RESULTS: In tumor cells, the high expression rates of BSP, OPN (30.0%,10.0%, respectively) in fibroblastic osteosarcoma were significantly lower than those of osteoblastic group (75.0%,64.3%, respectively). Other high expression rates of OC, BSP, OPN in or among different groups had no significant difference. In unmineralized neoplastic bone matrix OC, BSP, OPN were weakly expressed. In mineralized neoplastic bone matrix the high positive rates of BSP,OPN (81.4%,76.7% ) were significantly higher than that of OC(4.6% ). But OC was expressed strongly in mature bone. CONCLUSION: In tumor cells of the osteosarcoma expression of OC,BSP,OPN were accompanied by the cell differentiation. It seemed that during the formation and mineralization of neoplatic bone, BSP,OPN,OC had a relatively close relations with mineralization and they showed some extent sequensed expression.

[Ultramicrostructural observation on epithelial cell disdifferentiation in rat tongue carcinogenesis induced by 4NQO].

OBJECTIVE: To clarify epithelial cell ultramicrostructure changes in oral carcinogenesis. METHODS: The epithelial cell ultramicrostructures in tongue carcinogenesis induced by 4-nitroquinoline-1-oxide (4NQO) were observed by transmission electron microscope. RESULTS: With the progress of oral carcinogenesis, regular changes of epithelial cell were showed: decrease in tonofibril, keratohyaline granules, desmosome; and increase in mitochondria. The basement membrane was broken in some severe dysplasia (sDP) and was broken through by parts of cell processes in in situ carcinoma (ISC). CONCLUSION: Oral carcinogenesis is a multistep process. Epithelial cell aberrant differentiation is showed by a decrease in the synthetic products of epithelium cell.

[Apoptosis of Tca8113 cells induced by cisplatin and their cell cycle specificity].

OBJECTIVE: Evaluating the effect of apoptosis induced by cisplatin in human oral squamous cell carcinoma cells and their cycle specificity. METHODS: Hu man tongue squamous cell carcinoma cell line Tca8113 was used in vitro as a study object. Morphological features of apoptotic cell induced by cisplatin were observed by using of microscope, fluorescence microscope and transmission electronic microscope (TEM). The apoptotic rates were analyzed by DNA fluorescence staining assay and TEM observation. The cell cycle specificity was measured by using flow cytometry. RESULTS: The cell proliferation of Tca8113 cells was inhibited by cisplatin and their proliferation index decreased. Cells treated with cisplatin became smaller, rounded and then disattached. Red nuclear fragmentation with condensed chromatin was observed under fluorescence staining assay. Condensation and crack of nucleus and apoptotic bodies appeared in apoptotic Tca8113 cells observed under TEM. Cell cycle of Tca8113 cells was apparently arrested at S by cisplatin. CONCLUSION: Apoptosis was induced by cisplatin in Tca8113 cells, and it may be an important mechanism of SCC killed by cisplatin.

[Experimental study of abnormal cell proliferation and differentiation in rat tongue carcinogenesis induced by 4NQO].

OBJECTIVE: To clarify the phenotypic characteristics of abnormal cell proliferation and differentiation in oral carcinogenesis. METHODS: The expressions of gp230, a marker of epithelium cell differentiation, and proliferating cell nuclear antigen (PCNA) in rat tongue carcinogenesis induced by 4-nitroquinoline-1-oxide (4NQO) were detected by LSAB immunohistochemical staining. RESULTS: With the progress of oral carcinogenesis abnormal proliferation and differentiation patterns were indicated by reduced expression of gp230 and increased expression of PCNA. CONCLUSION: Oral carcinogenesis is the result of mucosa epithelium cell aberrant proliferation and differentiation.

[The Inhibitory Study of TNP 470 Combined CDDP on the Tumor Growth and Lung Metastasis of Adenoid Cystic Carcinoma ACC M in Nude Mice]

OBJECTIVE:To study the effect of TNP-470 combined CDDP on the tumor growth and lung metastasis of cell strain ACC-M in vivo. METHODS:High potential lung metastatic adenoid cystic carcinoma cell strains (ACC-M) 5x10(5) were injected simultaneously to 18 nude mice subcutaneously and enously. The animals were randomly divided into 3 groups. In group 1, TNP-470 was administered subcutaneously at the dose of 30mg/kg every other day for 2 weeks from the beginning of 3(rd) week to the end of 4(th) week. In group 2, 5mg/kg CDDP were injected into peritoneal cavity at the beginning of 3(rd) week and 4(th) week and TNP-470 in the same manner as group 1 additionally. In group 3, the same volume of 0.97% NaCl was injected in the manner of group 1. The mice were sacrificed by dislocating the neck at the end of 4 weeks. The tumor size and weights were recorded. The vascular density was measured by rabbit-anti-human factor VIII antibody stain. The results were analyzed by Student t and X(2) test. RESULTS:Group 1 and group 2 had fewer tumor weight and lung metastasis than the 3(rd) group. There was significant difference in the tumor weight and lung metastasis between group 1 and group 2. But the vascular density had no difference among three groups. CONCLUSION:TNP-470 combined CDDP has stronger inhibition on both the tumor growth and lung metastasis of human adenoid cystic carcinoma ACC-M than used alone.

[The in vitro comparison of platelet induced ACC-M cell adhesion with and without Aspirin]

ObJECTIVE:Observation the affection of aspirin to adhesion of highly metastatic salivary adenoid cystic carcinoma(ACC-M) cell induced by platelet.METHODS:Adhesive ratios of ACC-M cell induced by platelet with or without aspirin were measured.RESULTS:After the addition of platelet rich plasma(PRP),adhesive ratio of ACC-M cell increased significantly (P<0.01). The treatment with aspirin after addition of PRP led to a nonsignificant increase of adhesive ratio (P>0.05).The reduction of adhesive ratio by aspirin was found dosage independent within concentration range of 0.01-0.25g/L of aspirin. CONCLUSION:Adhesion of ACC-M cell was raised by added platelet.Addition of aspirin,platelet induced adhesion of ACC-M cell was inhibited significantly.

Effects of all-trans retinoic acid and interferon-gamma on expression of RAR beta gene in Tca8113 cells.

OBJECTIVE: All-trans-retinoic acid (ATRA) and interferons (IFNs) have been proven to synergistically amplify growth inhibition and apoptosis in the tongue squamous carcinoma cell line (Tca8113). Nuclear retinoic acid receptor-beta (RAR beta) was the key gene that mediated retinoid activity for squamous carcinoma cells. In order to understand the mechanism of ATRA combined with IFN gamma inhibiting growth of Tca8113 cells, this investigation focused on RAR beta mRNA expression. MATERIALS AND METHODS: In this experiment, RT-PCR method was used to analyze the RAR beta expression level, and viable cell count assay was carried out for growth inhibition studies. RESULTS: All-trans-retinoic acid (1 microM) and IFN gamma (1000 u/mL) inhibited cell growth by 39.2% and 44.4%, respectively. Synergistic inhibition of cell proliferation by 86.7% was found under combined treatment. The combination of suboptimal concentrations of ATRA (0.1 microM) with IFN gamma (1000 u/mL) showed a much stronger additive inhibitory effect on cell proliferation. ATRA (1 microM) and IFN gamma (1000 u/mL) increased RAR beta expression to 4 times and 3 times, respectively. The expression of RAR beta increased 12 times after treatment with combined ATRA and IFN gamma treatment. CONCLUSIONS: These observations indicated that the use of combined ATRA and IFN may lead to enhanced antitumor effects. These results also suggested that ATRA and IFN mediated upregulating expression of RAR beta may play an important role in synergistic inhibition of proliferation in Tca8113 cells.

[Expression of c-erb B-2 concoprotein in salivary adenoid cystic carcinoma]

OBJECTIVE:In order to analyze the correlation between immunohistochemical expressin for c-erb B-2 oncoprotein and salivary adenoid cystic carcinoma (ACC). METHODS: 25 cases of ACC and two ACC cell strain was studied immunohistochemically using a monoclone antibody against c-erb B-2 oncoprotein. RESULTS: No positive results were obtained in any cell strain,but the invasive breast cancer expressed strong positive. CONCLUSION: It demonstrated that c-erb B-2 expression can not be used as a predictive marker for prognosis of ACC.

Selection of adenoid cystic carcinoma cell clone highly metastatic to the lung: an experimental study.

Lung metastasis is very common in patients with adenoid cystic carcinoma (ACC). The purpose of the present study is to establish a model for an experimental study of ACC metastatic mechanisms and antimetastatic procedures. After five repeated selection in vivo, combined with an in vitro cloning technique and analysis of platelet aggregation activity, a clone (Acc-M) highly metastatic to the lung was selected from an ACC cell line (Acc-2). The metastatic rate was 96% vs 18% for Acc-M and the Acc-2 cell line. The metastatic rate and the weight of the lung with metastases correlated positively with platelet aggregation activity. The present study might be important for research into mechanisms of ACC metastases.

[The effect of FN(fibronectin) on invasion and metastasis of human acinic cell carcinoma cell strain of the salivary gland]

FN distribution in ACC-2 and high lung-metastatic ACC-M cells was studied with immunohistochemical staining.There were less FN found in ACC-2,but much more FN were observed in the highly metastatic clone Acc-M.In vitro motility of two cell lines was measured by Boyden chamber.With the presence of exgenous FN,Acc-2 became activated with a higher migration rate.The cell adhesion test showed that Acc-2 had stronger ability of adhesion than Acc-M with the presence of FN.The result suggested FN may play a vital role in the invasion and metastasis of ACC.

[Experiment study on biological and immunological characteristics of modified DNL of oral squamous cell carcinoma with TNF-alpha gene]

Growing characteristic of gene-transduced and nontransduced DNL in medium of containing rIL-2 was compared,and growing characteristic of gene-transduced DNL in medium of non containing rIL-2 was analyzed.DNA index (DI),cell cycle and immunologic phenotypes of gene-transduced and nontransduced DNL were analyzed with FACS.The results revealed interfered with growth,DL,cell cycle and immunologic phenotypes of DNL,and gene-transduced DNL couldn't growing infinitely.