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Objective: To investigate the growth of the tissue engineered mucosa after its hetero-transplantation., Methods: The epithelial cells and fibroblasts were isolated from a postoperative tissue of the 3-month patient with labial cleft. The epithelial cells and fibroblasts were separately seeded on the polylactic/glycolic acid copolymer membrane, and then they were exposed to the air-liquid interface. Seven volunteer patients, whose traumatic beds were repaired with the tissue engineered oral mucosa. The biopsy tissue from one of the seven patients was observed under light microscope 18 and 30 days after transplantation, respectively., Results: The tissue engineered oral mucosa having 5-6 layers anti-cytokeratin staining positively cells in the epithelial layer and 3-7 layers anti-Cytokeratin staining negatively cells in the subepithelial layer grew well after the hetero-transplantation. No difference could be found between the transplanting and normal areas. At 18 days, the epithelial layer and lamina propria grew well and the fibroblasts were found; at 30 days, collagen was obviously observed. The structures in both the transplanting and the normal areas were similar., Conclusion: The tissue engineered oral mucosa can grow well after the hetero-transplantation.

Objective: To evaluate the advantages and disadvantages of vascularized free peroneal composite flaps for reconstruction of oral and maxillofacial defects., Methods: From November 1999 to December 2002, 28 cases of oral maxillofacial defects were reconstructed with vascularized free peroneal composite flaps, with fibula-cutaneous flap in 21 cases and with fibula-myocutaneous flap in 7 cases. Three cases received insertion of dental implants into the fibula flap. The flap size was 3.0 cm x 5.5 cm to 8.0 cm x 12.0 cm; the fibula length was 5.5 cm to 16.0 cm., Results: Of the 28 flaps reconstructed, 24 survived, 3 necrosed partially and 1 necrosed completely. All the 5 implants survived and achieved good bone integration in 3 cases. Twenty-six cases were followed up 1-36 months with an average of 18.5 months, the facial appearance and the vocal function were satisfactory in 23 cases., Conclusion: Vascularized peroneal flap has many advantages and is one of the optimal flaps for reconstruction of oral maxillofacial defects.

Objective: The aim of this study was to investigate the delivery of Cucurbitacin BE to cervical lymph nodes by peri-cancer submucosal injection of the average diameter 85 nm Cucurbitacin BE polylactic acid nanoparticles (CuBE-PLA-NP)., Methods: Cervical lymph node metastasis model of mice oral cancer was established and CuBE-PLA-NP and CuBE were injected in peri-cancer submucosal of the mice respectively, the concentration in blood, heart, liver, spleen, lung, kidney and cervical lymph node were measured by high performance liquid chromatography at fifteen time points, Targeted cervical lymph nodes were evaluated by targeting index, selectivity index, targeting efficiency, relative targeting efficiency and ratio of maximum concentration., Results: (1) The CuBE concentration in the cervical lymph nodes after CuBE-PLA-NP injection was far higher than that after CuBE injection at every time point, the CuBE duration in the cervical lymph nodes in CuBE-PLA-NP group was far longer than that in CuBE group; (2) The five targeted values of the cervical lymph nodes in CuBE-PLA-NP group was far higher than that in CuBE group; (3) The CuBE concentration in blood, heart, liver, spleen, lung and kidney in CuBE-PLA-NP group was far lower than that in CuBE group., Conclusion: The CuBE-PLA-NP can specifically deliver CuBE to the cervical lymph nodes by peri-cancer submucosal injection, increase CuBE concentration and duration in metastasized cervical lymph nodes, and decrease drug concentration of other organs.

Objective: The aim of this study was to investigate the target delivery of Cucurbitacin BE (CuBE) to cervical lymph nodes by peri-oral-cancer submucosal injection of the average diameter 85 nm Cucurbitacin BE poly-lactic acid nanoparticles (CuBE-PLA-NP) and evaluate its clinical therapy efficacy., Methods: CuBE and CuBE-PLA-NP were respectively injected into peri-oral-cancer submucosa at 2, 12, 24, 48, 72, 96, 120, 144, 168 and 192 hours before operation in 26 patients with oral cancer. The concentrations of CuBE in cervical lymph nodes and blood were measured by high performance liquid chromatography (HPLC) at every time point. Cancer cell degeneration and necrosis in metastasis foci of cervical lymph nodes were observed using light microscope and electron microscope., Results: (1) The CuBE concentrations in cervical lymph nodes after CuBE-PLA-NP injection were far higher than those after CuBE injection at every time point, the duration of CuBE existing in the cervical lymph nodes in CuBE-PLA-NP group was far longer than those in CuBE group, the area under the CuBE concentrations-time curve of the cervical lymph nodes in CuBE-PLA-NP group was 43.67 times as many as that in CuBE group; (2) The CuBE ratio of maximum concentrations in the cervical lymph nodes in CuBE-PLA-NP group was 106.46 times as high as that in CuBE group; (3) The CuBE concentrations in the blood in CuBE-PLA-NP group was far lower than that in CuBE group; (4) Cancer cell necrosis and degeneration in the metastasis foci of cervical lymph nodes were found in CuBE-PLA-NP group, necrosis and degeneration were not found in CuBE group., Conclusion: The peri-oral-cancer submucosa injection of the average diameter of 85 nm CuBE-PLA-NP can specifically delivery CuBE to the cervical lymph nodes, enhance treatment efficiency, and reduce general toxicity.

Objective: The purpose of this article is to discuss the reconstruction of facial muscle defects with tissue engineered myoblast in SD rats., Methods: Using purified, subcultured myoblast of neonatal rats and type I collagen gels as extracellular matrix (ECM) and scaffold, tissue engineered muscle was transplanted in the face of syngeneic nutured rats., Results: Tissue engineered myoblast generated and differentiated in vitro were observed with microscope. Myoblast fused each other and formed myofibers in the face of rats, nerve fibers and vessels regeneration could also be found in some samples. Postoperative electromiographs showed the myofibers were active when stimulating the nerve trunk that innervates the engineered facial muscle., Conclusion: Tissue engineered method is hopeful to be used as a new technique to reconstruct defects of facial muscle.

Objective: To transfect nm23-H1 into the Acc-M cell lines in a safe, high-efficiency and low-toxicity way, and then to find out whether nm23-H1 affects the metastasis ability and chemo-sensitivity of Acc-M cell lines., Methods: Lipofect was used to transfect nm23-H1 into Acc-M cell lines. The difference in expression of nm23-H1 between the transfected and non-transfected cell lines was detected by immunohistochemistry. Then by use of transwell-room and wash way, the difference in invasion and metastasis ability between the transfected and non-transfected cell lines was tested. MTT method was adopted in finding the change of chemo-sensitivity., Results: Using pCMV-Neo-Bam system, we observed the stable expression of nm23-H1 and the significant difference in Nucleoside Diphosphate kinase-A, expression between the transfected and non-transfected Acc-M cell lines. The metastasis ability of transfected Acc-M cell lines decreased significantly. The chemo-sensitivity of transfected Acc-M cell line to cis-diamminedichloroplatin (C-DDP) increased significantly., Conclusion: nm23-H1 can inhibit the metastases of Acc-M cell lines significantly and can increase the chemo-sensitivity to C-DDP significantly.

Objective: The purpose of this study was to offer pathological reasons for partial parotidectomy adopted in parotid pleomorphic adenoma treatment., Methods: Continuous slices from 25 cases of primary parotid pleomorphic adenoma were carefully examined and the extra-envelope infiltration, the budding and the primary multi-center were measured and recorded., Results: The range of extra-envelope infiltration and budding was limited in 0.09-0.285 mm out of tumor capsules, which was far less than the safe boundary(1 cm) of the partial parotidectomy. Two cases with multi-centers were found, but all the centers were aggregated together and encapsulated in a united envelope., Conclusion: Partial parotid parotidectomy is a safe method and can remove the tumor completely in parotid plemorphic adenoma treatment.

Objective: To evaluate whether the vascular endothelial growth factor (VEGF)-C cDNA which cloned from a patient with squamous cell carcinoma (SCC) of tongue can encode a functional protein or not., Methods: RT-PCR was employed to clone the functional VEGF-C fragment from the surgical specimen of a lingual SCC patient. Then it was subcloned into expressive plasmid vector pBKCMV, which was transfected into E. coli to examine its expression., Results: A truncated human VEGF-C cDNA fragment was amplified from the lingual SCC. The sequencing results of the fragment demonstrated that it had 99.6% similarity with the reported human VEGF-C cDNA (representing the 559-1611 bp according the sequence of Genbank Entry X94216). Induced with IPTG, the E. coli XLI-Blue MRF' containing the recombinant pBK-VEGF-C expressed a 56,000 fusion protein, which can be recognized by polyclonic anti-human VEGF-C antibody., Conclusion: A functional fragment VEGF-C cDNA was cloned from a lingual SCC. It will promote more intensive research on the function of VEGF-C and its relationship with metastasis of oral SCC.

Objective: To study the regulation and effect of the expression of nm23-H1 gene in different processes of regional lymph node metastases of oral squamous cell carcinomas(OSCC)., Methods: By immunohistochemical analysis in 200 paraffin-embedded tissues of OSCC and Western Blot in 9 fresh tissues of OSCC using a monoclonal antibody, the expression of nm23-H1/NDPK-A were detected in dividing groups accompanied with the clinical and pathological data of cervical lymph node metastases and modes of invasion., Results: The rates of negative expression of nm23-H1/NDPK-A had significant differences between metastatic cases(34/81) and non-metastatic cases(15/119). The nm23-H1/NDPK-A negative group showed higher frequency of lymph node metasteses (P < 0.01). In the different metastatic processes, There were significantly differential expressions of nm23-H1/NDPK-A among stage N0, N1 and N2(P < 0.01), distinct involved quantities of lymph nodes (P < 0.01), dissimilar metastatic involved levels(P < 0.05), definite modes of invasion(P < 0.01) and different cell differentiations(P < 0.01). There were much higher negative expressions in those cases of N1 stage(23/38), only one lymph node to be involved(23/39), metastasized to submandibular nodes or/and superior deep cervical nodes(17/28 and 12/29, respectively), IV c types of invasion(33/54) and poor differentiations(9/19)., Conclusion: The results indicated that nm23-H1/NDPK-A played an more important role in switching the initial metastases formation than in influencing the later metastases spread because the negative expressions of nm23-H1/NDPK-A in solid tumors of OSCC would induce the formation of high metastatic cellular subpopulations. It was also confirmed that nm23-H1 gene might be a metastatic suppressor and may be useful in predicting the initial lymph node metastases in OSCC.

Objective: A functional VEGF-C cDNA was cloned from a patient with squamous cell carcinoma (SCC) of tongue in order to study the important role of vascular endothelial growth factor (VEGF)-C in lymphatic dissemination of malignancies in gene level., Methods: RT-PCR was employed to clone the human VECF-C encoded cDNA from a surgical specimen of a lingual SCC patient. Then it was subcloned into plasmid vector pCRII and sequenced., Results: A 1.1 kb human VEGF-C cDNA fragment was amplified from the lingual SCC. The sequencing results of the fragment demonstrated that it had 99.6% similarity with the reported human VEGF-C cDNA (representing the 559-1,611 bp according the sequence of Genbank Entry X94216)., Conclusion: An encoded fragment VEGF-C cDNA was successfully cloned from a lingual SCC and provided a necessary material for further study.

Objective: To study the role that vascular endothelial growth factor-C plays in the lymph node metastasis of the squamous cell carcinoma (SCC) of tongue., Methods: An established VEGF-C transfected Tca8113 cell strain (VcTca) was heterotransplanted into BALB/C nude mice subcutaneously. The expression of VEGF-C and VEGFR-3 was detected. The property of tumorigenesis and metastasis of the transfected cell was investigated., Results: Jugular lymphatic metastases developed in 4/6 mice of VcTca group in 6 weeks after transplantation, while no lymph node metastasis was found in mice with tumors of non-transfected Tca8113 cell (Tca) and mock transfected cell(pDTca) at the same time. There was no difference in tumor weight among VcTca, pDTca and Tca (P > 0.05)., Conclusion: It is suggested that VEGF-C could promote lymph node metastatic ability of Tca8113 cell, and could be utilized as a target of new regimen in prevention and treatment for lymphatic metastasis of oral carcinomas.

Objective: The aim of this study was to examine the effects of deep branches of cervical plexus on trapezius muscle., Methods: The evoked electromyography of 32 trapezius of 16 SD rats was examined in different time and results obtained were processed by statistical methods., Results: After cutting the accessory nerves, stimulating the deep branches of cervical plexus evoked the myoelectricity, the myoelectricity results varied according to different time., Conclusion: The deep branches of cervical plexus are another important motorial supply to trapezius muscles. After cutting the accessory nerves, if the branches were undamaged, the shoulder's function would be greatly preserved.

Objective: The purpose of this study was to investigate the cisplatin release in peripheral blood and tumor tissue after cisplatin-loaded albumin microspheres (CDDP-AMS) were infused through lingual artery for treating carcinoma of tongue., Methods: Anticancer agent cisplatin was encapsulated with biodegradable and non-antigenic albumin. The cisplatin-loaded albumin microspheres, which were 56.3 microns in diameter, were used for treating 7 patients with squamous carcinoma of tongue by infusing through lingual artery. The cisplatin in peripheral venous blood and carcinoma tissue were measured for analyzing the drug release in vivo., Results: Concentration of the drug in peripheral venous blood increased to the highest level in 90 minutes after CDDP-AMS was infused through lingual arterial and still could be detected until 10th day. The cisplatin concentration in tumor tissue kept considerably stable and could be detected until 17 days., Conclusion: CDDP-AMS had the effect of controlled release in vive. Effective and steady local drug concentration played the role of killing tumor cells, the low concentration in peripheral blood reduced the side effects of cisplatin to minimum as well. It is concluded that CDDP-AMS is a useful and recommendable method for treating the carcinoma of tongue.

Objective: The aim of this study was to investigate the effects of preserving external jugular vein and cervical plexus in reducing intra-cranial pressure, face swelling as well as in avoiding shoulder arm syndrome in neck dissection., Methods: 20 patients with oral cancer were divided into two groups. The facial interstitial pressure was examined before and after neck dissection and lymph tubes were stained by immunochemical techniques., Results: The degree and lasting time of the increase of facial interstitial pressure in group with external jugular vein and cervical plexus preserved was lower than that in control group. The density of lymphatic tubes around the internal jugular vein, accessory nerve and sternocleidomastoid muscles was much higher than that around the external jugular vein and cervical plexus., Conclusion: Preserving external jugular and cervical plexus during radical treatment is effective not only in reducing the intra-cranial pressure and face swelling, but also in preventing shoulder arm syndrome.

Objective: The purposes of this study were to clone and sequence the major histocompatibility complex type I (MHC I) molecular antigen recognizing gene (H-2Kk) of 615 mice, and to provide the functional gene for transgenic therapy., Methods: The 1.4 kb full-length fragment of H-2Kk gene complementary DNA (cDNA) was amplified from the total RNA of 615 mouse liver by using reverse transcription polymerase chain reaction (RT-PCR). The cDNA was inserted into PGEM3Zf(+) vector directionally, and the competent E. coli JM109 was transformed with the ligated product. The recombinant PGEM3Zf(+)-H-2Kk cDNA plasmid was obtained using restricted enzyme analysis of the transfectants. The complete sequence of 615 mouse H-2Kk cDNA was determined by using Sanger's method., Results: The sequences of 615 mouse H-2Kk cDNA were 99% similar with those of H-2Kk cDNA which were reported by other researchers, and the sequences encoding antigen recognizing regions (ARS) were identical with each other., Conclusion: The authors cloned the MHC I molecular antigen recognizing gene (H-2Kk) of 615 mice successfully and got the functional gene of MHC I.

Objective: The purpose of this study was to evaluate the advantages and disadvantages of vascularized free fibula flap as a new method for mandibular reconstruction., Methods: 25 cases (17 male to 8 female) who have received mandibular reconstruction with free vascularized fibular flaps in our hospital were studied retrospectively. The average length of the fibula grafts is 10.0 cm (range from 5.5 to 16 cm). 3 cases received primary insertion of osteointegrated dental implants into the free fibula flap, and all these 5 implants survived., Results: All flaps except 1 were viable. 62% of the cases took normal diet postoperatively, and the remainder took soft diet as well. All patients spoke clearly. No ankle unstability was reported. And the aesthetic assessments in all patients were good or fair., Conclusion: Vascularized free fibular flap takes its distinct advantages to other autogeneous free bone flaps and is confirmed to be one of the optimal methods for mandible reconstruction by our study.

Objective: The purpose of this study was to investigate the indication and therapeutic effects of Pingyangmycin injection as a primary therapy of lymphangiomas in oral, maxillofacial and cervical region., Methods: A total of 195 patients (106 males and 89 females) with lymphangiomas in oral and maxillofacial regions were treated in the affiliated dental hospital of Sichuan University from May 1990 to December 2000. The patients' ages ranged from 0.5 to 46 years. The tongue was the most commonly involved site, followed by the cheek and the neck. The 200 lymphangiomas (5 patients had 2 lymphangiomas in different sites) underwent the therapy of Pingyangmycin, which was injected as with 1 mg/ml in saline. The total dose of Pingyangmycin ranged from 5 mg to 70 mg and 5 to 58 times, 1 time per 2-4 weeks., Results: The curative rate of cystic-type lymphangiomas was the highest. Of the 51 cystic lymphangiomas, 110 capillary lymphangiomas, 18 cavernous lymphangiomas and 21 combinations of capillary and cavenous lymphangiomas, the curative rates were respectively 100% (51), 46.36% (51), 16.16% (3) and 19.05% (4), which showed a significant therapeutic effect, respectively. And 40(78.43%), 19(17.27%), 2(11.11%) and 0(0%) of them completely disappeared. There was no serious side effect with Pingyangmycin-injection treatment, such as pulmonary fibrosis., Conclusion: The treatment of injection of Pingyangmycin is a selective primary method of lymphangiomas, which can reduce the size of lymphangiomas, and make them completely disappeared.