[Application of team approach and key techniques of cleft lip and palate].

The development of an expert consensus based on specific domestic situations will provide practical guidance to the efforts aiming at improving cleft care in China. The team approach of twenty-one cleft centers were pooled together, covering pre-surgical orthopedics, primary surgical repair, orthodontic treatment, alveolar bone graft, secondary deformity correction, palatal fistulae repair, the diagnosis and treatment of velopharyngeal incompetence, speech therapy, otitis media management, and skeletal deformity correction. Agreement was achieved among the authors concerning the application of critical surgical and non-surgical techniques. The ambition of this consensus is to introduce more clinicians to the revolution of sequential treatment of clefts, and form the basis for a more comprehensive cleft care manual in the future.

Study on self-microemulsifying membrane controlled-release drop pill of hawthorn leaves flavonoids / 中国中药杂志

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.

Clinical study of submental artery island myocutaneous flap for reconstruction of oral and maxillofacial defects following operation / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the outcome and indication of the reconstruction of oral and maxillofacial postoperative defects by submental artery island myocutaneous flaps.</p><p><b>METHODS</b>Sixty eight cases with the reconstruction of oral and maxillofacial defects by submental artery island myocutaneous flaps from January 2006 to May 2010 were analysed retrospectively. Primary lesions included carcinomas originating from tongue (28 cases), palate (13 cases), mouth floor (9 cases), gingiva (4 cases), buccal mucosa (6 cases), lip (3 cases), and other malignant or benign tumors (5 cases). The ages ranged from 25 to 84 years (mean 58 years); 47 males and 21 females. The sizes of skin paddle varied from a minimum of 4 cm × 4 cm to a maximum of 15 cm × 10 cm.</p><p><b>RESULTS</b>Of the 68 flaps, 62 were survival, 4 had partial necrosis but healed with treatments, and 2 failed due to complete necrosis. Appearance and functions of recipient sites were satisfactory. The followed-up time was 3 - 24 months, local recurrence occurred in 5 cases and cervical lymph node metastases were found in 15 patients.</p><p><b>CONCLUSION</b>Submental island flap is reliable for the reconstruction of postoperative defects in early oral cancer without regional lymph node metastasis or in benign tumor.</p>

Suppression to ameloblastoma xenografts of chicken embryo chorioallantoic membrane by tissue inhibitor of metalloproteinases-2 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane (CAM) after tissue inhibitor of metalloproteinase-2 (TIMP-2) gene transfection.</p><p><b>METHODS</b>Fresh ameloblastoma tissues were minced into 1-2 mm3 and transplanted on the CAM. There were three groups named as control group (Empt), plasma transfection group (Lipo), and TIMP-2 gene transfection group (P). The specimens were respectively investigated by microscope indifferent spots after implanting. The volume of the xenografts and the weight of xenografts in the termination time of the experiment were recorded. The invasiveness of xenografts was divided into four grades by pathological examination. Western blot analysis was performed to investigate matrix metalloproteinase-2 (MIMP-2) and TIMP-2 protein in xenografts.</p><p><b>RESULTS</b>Ameloblastoma tissues can survive on CAM and the tumor cells may invade it on 5-7 days after implanting. At 9 d after implanting, the invasiveness grades in P group were 7 in grade 0, 1 in grade 2, 0 in grade 3. The expression of TIMP-2 protein in P group was significantly higher than that in Empt group (P < 0.05). The expression of MMP-2 protein in P group was lower than that in Empt group (P < 0.05).</p><p><b>CONCLUSION</b>The xenotransplanted tumor model of human ameloblastoma on CAM was successfully established. The invasiveness of ameloblastoma xenografts was suppressed might be due to TIMP-2 gene transfection.</p>

Application of multislice spiral CT and 3D reconstruction in diagnosis and treatment of vascular malformations in head and neck / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the application of MS-CT and 3D reconstruction in diagnosis and treatment of vascular malformations in head and neck.</p><p><b>METHODS</b>20 cases with vascular malformations in head and neck underwent MS-CT and 3-D reconstruction. Then the treatment was determined based on the results of MSCT scanning. The postoperative results were evaluated.</p><p><b>RESULTS</b>The images of MS-CT showed the edge of vascular malformations partially or completely in 16 cases of venous malformations. The lesion's anatomic site and 3-D position was obtained. The 3-D images also showed the overexpanded supply arteries in 4 case of arteriovenous malformations. 2 case of venous malformations in lip underwent resection and healed completely. 12 cases of venous malformations in buccal and floor of mouth were treated with compartmentalized sclerotherapy with partial lesion involution. 2 case of venous malformations in mouth floor were treated with operation followed by sclerotherpy with partial lesion involution. 4 cases of arteriovenous malformations were treated with Superselective Artery Embolization with partial lesion involution.</p><p><b>CONCLUSION</b>MS-CT and 3D reconstruction can play an important role in diagnosis and treatment of vascular malformations in head and neck.</p>

A comparison of endoscopy-assisted and conventional partial-superficial parotidectomy / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of endoscopy-assisted partial-superficial parotidectomy.</p><p><b>METHODS</b>38 cases with benign tumors located in the superficial lobe of the parotid gland were randomly assigned to receive conventional (20 cases) or endoscopic (18 cases) partial-superficial parotidectomy. Two short incisions, which were 2 to approximately 2.5 cm in length and located at retromandibular and postauricular area, were adopted for endoscopy-assisted surgery. The facial nerve was dissected through retrograde approach.</p><p><b>RESULTS</b>The tumors were successfully resected with endoscopy in 18 cases. The operation time was not significantly different between the conventional and endoscopy-assisted procedures (P > 0.05). The intraoperative blood loss was markedly lower in endoscopy-assisted group, compared with conventional group (P < 0.01). All the 18 cases with endoscopy-assisted surgery were satisfactory with the postoperative cosmetic results. The great auricular nerve was preserved very well in 12 patients (66.6%). Transient facial paralysis happened in 1 case and relieved 1 month later. Salivary fistula occurred in 1 case and recovered after dressing with pressure for 2 weeks. All the patients were followed up for 24 to approximately 50 months (mean, 39 months) without relapse.</p><p><b>CONCLUSIONS</b>Endoscopy-assisted partial-superficial parotidectomy can successfully treat benign tumors located in the superficial lobe of parotid gland with a better postoperative cosmetic result.</p>

Evaluation of the articulator function in the hemi-tongue defect patients after radical tongue cancer resection and simultaneous reconstruction with forearm flap or primary close / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the effect of the patients' articulator function after reconstruction of hemi-tongue defect with forearm flap (FAP) or prime close (PC).</p><p><b>METHODS</b>36 patients who underwent hemiglossectomy were investigated after radical surgery for TSCC. 20 cases were reconstructed with FAP flaps and 16 with primary closure. The patients' articulator functions were evaluated by articulation tests. VS-9700 was used to analyze the speech character when they pronounce /ji/.</p><p><b>RESULTS</b>1) The speech articulation of patients who underwent hemi-tongue reconstruction with FAF was better than that of patients with PC, and there was significant difference between them (P < 0.05). 2) The first formant (F1) of /i/ of the PC group was lower than that of the control group (P < 0.05). But the second formant (F2) of the PC group was higher than that of the control group (P < 0.05). The first formant (F1) of /i/ of the FAF group was lower than that of the control group (P < 0.05), but there were no significant differences between FAF group and control group in F2 of /i/ (P > 0.05).</p><p><b>CONCLUSIONS</b>Articulator function can be well achieved by forearm flaps reconstruction to hemi-tongue defect patients.</p>

Silencing of survivin gene enhances chemosensitivity of human tongue cancer cell line Tca8113 to cisplatin / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of survivin short hairpin RNA (shRNA) on survivin expression, cell apoptosis, and chemosensitivity of human tongue cancer cell Tca8113 to cisplatin.</p><p><b>METHODS</b>Survivin-directed shRNA plasmid vector was delivered into Tca8113 cells with lipofectamine(TM) 2000 reagent. Survivin expression was detected with the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Flow cytometry was used to examine cell apoptosis, and the sensitivity to anticancer agents was evaluated by methyl thiazolyl tetrazolium (MTT) assay.</p><p><b>RESULTS</b>After survivin shRNA vector transfection in Tca8113 cells, the expression of mRNA/protein declined significantly, and the apoptotic rate increased in time-dependent manner up to 37.9% at 48 hours. RNAi-mediated survivin reduction selectively inhibited growth and enhanced chemosensitivity of cisplatin but not of 5-fluorouracil.</p><p><b>CONCLUSIONS</b>Survivin shRNA could inhibit the expression of survivin mRNA and it's protein and enhance the chemosensitivity of cisplatin.</p>

Role of survivin gene on the apoptosis of Tca8113 cells induced by cisplatin / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the induction of apoptosis of cisplatin (DDP) to oral squamous cell carcinoma cell line (Tca8113) in vitro and study the role of Survivin on the apoptosis of Tca8113 cells induced by cisplatin.</p><p><b>METHODS</b>The inhibitory effects of different doses of DDP on Tca8113 cells were assayed with MTT test. Apoptosis was determined by flow cytometry. The expression of Survivin was detected by RT-PCR and immunocytochemistry.</p><p><b>RESULTS</b>Cisplatin obviously inhibited Tca8113 cells growth in a dose and time dependent manner. The apoptotic index showed the similar trend. Survivin gene expression was decreased with increasing of time and reached the lowest level at 24 hours after DDP treatment, then increased after that time.</p><p><b>CONCLUSION</b>Cisplatin gene can effectively induce apoptosis in Tca8113 cells and the inhibition of Survivin gene expression may play a critical role on Tca8113 cell apoptosis induced by cisplatin.</p>

Effects of guanine-quadruplexes formation induced by adriamycin on telomeric extension reaction mediated by telomerase of Tca8113 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of adramycin to disturb telomeric extention reaction mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form guanine-quadruplex (G4) structures.</p><p><b>METHODS</b>In the presence of adriamycin, d(TTAGGG)4, d(TTAGAG)4, d(TTAGGG)5 and d(TTAGGGT) were analyzed by electrophoretic mobility shift assay. The mobility of d(TTAGGG)3, d(TTAGGG)4 and d(TrAGGG)5 in native polyacrylamide electrophoresis were observed. Methylation protection experiments were performed to investigate the effects of adriamycin on methylation of guanine in d(TTAGGG)4 and d(TTAGAG)4. The traditional telomeric repeats amplification protocol (TRAP) and modified TRAP-G4 assays were, respectively, used to analyze the different characteristcs of adriamycin's inhibiting telomeric extension mediated by telomerase of Tca8113 cells.</p><p><b>RESULTS</b>At 5.00 microg/mL of adriamycin, conversion of some of linear d(TrAGGG)4 and d(TrAGGG)5to the new, high-mobility bands formed by complex with special second structures were found in the mobility shift assay. Adriamycin at 1.25 microg/mL protected the G in d(TIAGGG)4 from methylating. Adriamycin at 2.50 microg/mL or 1.25 microg/mL partially inhibited the telomeric extension lengthened by telomerase of Tca8113 cells in TRAP assay, but completely did so in TRAP-G4 assay.</p><p><b>CONCLUSION</b>Adriamycin is able to disturb telomeric extention mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form intra-molecular G4 structures.</p>

Establishment of subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To establish an subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice.</p><p><b>METHODS</b>Ameloblastoma cells were absorbed by primary culture, repeat attachment and pancreas proteolytic enzyme were both used to purify them. Then, the purified cells were implanted subcutaneously into the nude mice. The specimens were respectively investigated by microscope in different spots after implanting.</p><p><b>RESULTS</b>Ameloblastoma cells can survive in all of the 8 nude mice. The xenograft can be found on 23 days after implanting. The rate of successful inocutation is 25%. The subcutaneously xenotransplanted tumor cells can be found with microscope in the inter-muscle tissues of nude mice.</p><p><b>CONCLUSION</b>The subcutaneously xenotransplanted tumor model of human ameloblastoma in nude mice was successfully established and it may benefit to further studies on this tumor.</p>

Reconstruction of mandible defect in osteoradionecrosis patients with free fibula osteomyocutaneous flap / 中华显微外科杂志

Objective To explore the clinical applicating and efficacy of free fibula osteomyocutane- ous flap in mandible defect reconstruction in osteoradionecrosis patients.Methods The mandible defects were reconstructed by free fibula flaps with or without muscle cuff.The soft tissue defects were repaired by skin paddles.Status of osteotomy in fibula and flap survival was recorded.The complication in recipient site and donor site,as well as mouth opening and occlusion were reviewed.Facial contour and chewing function after reconstruction were evaluated.Results Patients were followed up 3-16 months.4 free fibula flaps with muscle cuff and 5 without muscle cuff survived well.The size of mandible defects covered from 6cm to 17cm. And the harvested fibula flaps with length of 8.6-17cm were cut into 3 segments in 2 cases,and 2 segments in 5 cases.Fibula flap was divided into 2 segments and overlapped in 2 cases.No serious complication was oh- served in recipient site and donor site.Satisfying esthetic result and normal occlusiong of heath mandible were obtained in all cases.The degree of mouth opening was 2.5-3.3cm.Fair chewing function was revealed in re- constructive region after prosthesia repaired.Conclusion Free fibula osteomyocutaneous flap is relatively ideal reconstruction material of mandible defect in osteoradionecrosis patients for its high survival rate and well esthetic results.

Intraoral submandibular gland excision and how to deal with external maxillary artery / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility and safety of intraoral submandibular gland excision.</p><p><b>METHODS</b>Analyze the relationship between the external maxillary artery and submandibular gland, and offer a reliable anatomical base for 10 cases of intraoral submandibular gland excision, including 8 cases of chronic sialadenitis, 1 case of pleomorphic adenoma and 1 case of cyst of submandibular gland.</p><p><b>RESULTS</b>The external maxillary artery went across the surface of gland submandibular, and its branches provided nutrition for the gland in most cases. The results of 10 cases intraoral submandibular gland excision were effective and satisfied, without major complications. The average time of operation was 50 minutes and the average hemorrhage of operation was 60 ml.</p><p><b>CONCLUSION</b>Intraoral submandibular gland excision is safe and feasible for chronic sialadenitis and cyst of submandibular gland and some of benign tumor submandibular gland as long as indications strictly controlled and the external maxillary artery well coped with.</p>

Construction and selection of siRNA expression cassettes targeting human telomerase reverse transcriptase gene in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To determine whether the human telomerase reverse transcriptase (hTERT) gene silencing could be effectively induced by PCR-derived siRNA expression cassettes (SEC) transfected by the fifth generation polyamidoamine dendrimer (G5 PAMAM-D) in Tca8113 cells.</p><p><b>METHODS</b>Four SEC were rationally designed and constructed based on a two-step PCR reaction. The SEC were then transferred into Tca8113 cells using G5 PAMAM-D, and hTERT expression was investigated by real-time fluorescence-quantitative reverse transcriptase-PCR and western blot analysis.</p><p><b>RESULTS</b>The RNA interference effects of the SEC targeted for varying hTERT mRNA positions showed a significant disparity. Among them, SEC-A revealed the most potent inhibitory effects (above 95% of reduction), followed by SEC-D and SEC-C, and SEC-B had no effect on hTERT expression (P > 0.05). That the endogenous hTERT gene silencing induced by G5 PAMAM dendrimer-mediated SEC-A was highly sequence-specific, and multiple transfection as well as properties of the vectors were routinely attributable to the specific suppression.</p><p><b>CONCLUSIONS</b>Specific inhibition of endogenous hTERT expression by use of a PCR-based short hairpin siRNA technique and dendrimer transfer system may serve as a novel strategy for treatment of tongue cancers expressing hTERT in vitro.</p>

Radiation-inducible promoters-mediated cdglytk gene in the treatment of buccal carcinoma in golden hamster / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of CDglyTK gene mediated by radiation-inducible promoters in the treatment of buccal carcinoma in Golden Hamster.</p><p><b>METHODS</b>Animal models of buccal carcinoma in golden hamster were established by painting 0.5% dimethyl-benzanthracene. The plasmids pcDNA (+) 3.1/E-CDglyTK were transfected into tumors by lipofectamine. 24 h later, the tumors were exposed to 3 Gy irradiation. Animals were monitored at regular intervals for volume of tumors. CDglyTK mRNA was assayed by RT-PCR. Apoptosis and proliferating cell nuclear antigen were detected respectively by in situ end-labeling and immunohistochemical methods.</p><p><b>RESULTS</b>Compared with control groups, the tumor was suppressed obviously by CDglyTK gene therapy combined with 3 Gy induction radiation. The expression of CDglyTK gene could be detected by RT-PCR in the transfected tumor, and up-regulation of CDglyTK expression was found in tumor exposed to radiation (P < 0.05). There was significant difference in apoptosis index or proliferation index between tumor without irradiation and tumor with irradiation (P < 0.05).</p><p><b>CONCLUSIONS</b>The radiation-inducible promoter can be served as a molecular switch to regulate the expression of CDglyTK gene in buccal carcinoma in golden hamster, and low dose induction radiation can significantly improve the therapeutic effects.</p>

Combined repair of large defect caused by radical surgery of advanced tongue cancer with rib-major pectoralis myocutaneous flap carrying costal parietal pleura / 中华外科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Association of matrix metalloproteinase-2 activity with cell proliferation and growth in ameloblastoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between matrix metalloproteinase-2 (MMP-2)activity and cell proliferation, growth and invasion of ameloblastoma.</p><p><b>METHODS</b>The cells and xenograft of ameloblastoma were treated with MMP-2 inhibitor Ro31-9790 and the effects of Ro31-9790 on the cell proliferation and growth of ameloblastoma were observed. Primary culture in vitro, subcapsular kidney xenograft in vivo, MTT assay, flow cytometry, neoplastic volume measurement and histochemistry were employed to study the effects of cell proliferation and growth produced by Ro31-9790.</p><p><b>RESULTS</b>There was no significant different in cell proliferation at same interval among several groups (P > 0.05). The ratio of G0/G1 stage, G2/M stage and apoptotic cells didn't increase following increased Ro31-9790, and the ratio of S stage cells also didn't reduce following increased Ro31-9790. The tumor volume and its increase in treatment group were significant less than those in control group.</p><p><b>CONCLUSION</b>Ro31-9790 does not influence proliferation of ameloblastoma cells in vitro, but it can effectively inhibit the ameloblastoma growth in vivo. MMP-2 activity has no relationship to proliferation of ameloblastoma cells, but it can contribute to the ameloblastoma growth and may be a reason of invasion in ameloblastoma.</p>

Synthetic radiation-inducible promoter mediated CDglyTK gene in treatment of Tca8113 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of CDglyTK gene mediated by synthetic radiation-inducible promoter in the treatment of Tca8113 cells.</p><p><b>METHODS</b>CDglyTK gene in pCEA-CDglyTK was subcloned into pcDNA3.1 (+) to construct plasmid pcDNA3.1 (+)-CDglyTK, and then the synthetic radiation-inducible promoter in pMD18 -T -E was inserted into pcDNA3.1 (+) -CDglyTK to construct plasmid pcDNA3.1 (+ )/E -CDglyTK. The recombinant plasmid was transfected into Tca8113 cells by lipofectamine, and then exposed to 3 Gy irradiation. Cytotoxicity was evaluated by MTT. The expression of CDglyTK gene was detected by RT-PCR. The apoptosis and proliferation were examined by flow cytomtery.</p><p><b>RESULTS</b>The plasmid pcDNA3.1 (+)/E-CDglyTK was constructed successfully. The comparative survival rate of Tca8113 cells was markedly decreased by induction irradiation. Up-regulation of CDglyTK expression was found in Tca8113 cells exposed to irradiation. The apoptosis index (AI) of Tca8113 cells exposed to irradiation was higher than that of Tca8113 cells without irradiation, the other way round, the proliferation index (PI) of Tca8113 cells exposed to irradiation was lower than that of Tca8113 cells without irradiation.</p><p><b>CONCLUSION</b>The synthetic radiation-inducible promoter can be served as a molecular switch to improve the expression of CDglyTK gene in Tca8113 cells, and low dose induction radiation can significantly improve the therapeutic efficiency.</p>

Effects of apoptosis of Tca8113 cells induced by adriamycin on telomerase and telomere repeat binding factor proteins / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the role of telomerase and telomere repeat binding factors (TRF) in apoptosis.</p><p><b>METHODS</b>The proliferative activity of Tca8113 cells was assessed by methyl thiazolyl tetrazolium (MTT) assay. After Tca8113 cells were treated with adriamycin at 5 mg/L, apoptotic morphology was observed under microscope with Giemsa staining and apoptosis examined by flow cytometry; analysis of telomerase activity was performed by TRAP-enzyme-linked immunosorbent assay; expression and expression level of TRF proteins were detected with immunohistochemical staining and immunofluorescence label assay, respectively.</p><p><b>RESULTS</b>After Tca8113 cells were treated with adriamycin at 5 mg/L for 5 days and 7 days, the cells apoptosis was found. Telomerase activity dropped in time-dependent manner. Expression of TRF proteins appeared in nucleus of the cells. No statistical difference in expression levels of TRF was observed between the treated and untreated cells.</p><p><b>CONCLUSIONS</b>Tca8113 cells apoptosis induced by adriamycin decreased telomerase activity, but did not influence the expression level of TRF proteins.</p>

The construction and expression of PcDNA3.1(+)/GFP-TIMP-2 in human ameloblastoma cell / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of TIMP-2 gene and to explore its expression in human ameloblastoma cell in vitro.</p><p><b>METHODS</b>The aimed gene fragment was obtained by RT-PCR. And then, molecmicrolar cloning technology and enzyme digestion were used to connect the gene with the plasmid PcDNA3.1(+), which can be expressed in eukaryotic cells and a report gene: green fluorescent protein gene (GFP) was already existed in the plasmid. We named the eukaryotic expression vector, which contended our aimed gene TIMP-2 as well as report gene GFP, PcDNA3.1(+)/GFP-TIMP-2. The vector was identified by PCR analysis, EcoR I and Xho I restriction analysis and Sequence analysis. After the PcDNA3.1(+)/GFP-TIMP-2 was transfected into cultured human ameloblastoma cell, RT-PCR and Flow Cytometry (FCM) and Microscope wre respectively performed to evaluate the effect of transfection and expression.</p><p><b>RESULTS</b>The constructed vector PcDNA3.1(+)/GFP-TIMP-2 was proved correct by enzyme digestion and sequencing analysis. After PcDNA3.1(+)/GFP-TIMP-2 was trasnfected into cultured human ameloblastoma cell, the rate of transfection is 47.6% (Analysis report of FCM), the green fluorescence was found in plasm (observed with fluo-microwave), the expression of TIMP-2 mRNA was elevated 2.4 times compared with the control group.</p><p><b>CONCLUSIONS</b>PcDNA3.1(+)/GFP-TIMP-2 was successfully constructed and it could be transfected into cultured human ameloblastoma cell. It may be benefit to further study of the relationship between the TIMP-2 gene and the behaviour of ameloblastoma.</p>