Evaluation of therapeutic effects of 125I particles brachytherapy for recurrent bladder cancer.

The aim of the present study was to evaluate the clinical value of 125I particles implantation in the treatment of recurrent bladder cancer. The study is a retrospective analysis of 32 patients with recurrent bladder cancer treated between May 2010 and January 2010. Of these, 16 cases (chemotherapy group) received conventional chemotherapy. A total of 16 patients (125I group) received radiotherapy with 125I particles, followed by conventional chemotherapy. By guidance of B ultrasound, 125I radioactive particles were implanted. All 32 patients were relieved after treatment, and the tumors were significantly reduced after 2 months. However, the tumors in the 125I group were significantly smaller than those in the chemotherapy group (P<0.05). The patients were followed-up for 1 year and no recurrence was found. Additionally, no complications occurred. Compared with the chemotherapy group, the tumor volume of the 125I group was significantly reduced (P<0.05). The disease-free survival and 5-year survival rates of the patients in the follow-up showed that the disease-free survival and 5-year survival rates of the patients in 125I group were significantly improved compared to those in the chemotherapy group. Therefore, the results have shown that 125I radioactive particles in the treatment of bladder cancer improve the symptoms of patients with bladder cancer in the short term, and continuously kill residual tumor and prevent recurrence.

[Effect of platelet rich fibrin combining with Bio-oss to treat furcation involvement].

PURPOSE: To evaluate the effect of platelet rich fibrin (PRF) combining with Bio-oss in treating Class II furcation involvement. METHODS: Thirty patients who had Class Ⅱ furcation involvement in the mandibular first molar were included. They were all free of systemic disease. After initial periodontal therapy, they were randomly divided into 2 groups. In the experimental group, PRF combining with Bio-oss were placed in the areas with furcation defect and covered with PRF. In the control group, only flap surgery was performed. All patients were followed up for 6 months after operation. The efficacy was evaluated with clinical parameters and cone-beam computed tomography (CBCT). The data were analyzed with SPSS 2.0 software package. RESULTS: Periodontal indexes including probing depth (PD), clinical attachment loss (CAL), horizontal probing depth (HPD) significantly decreased in both groups after operation (P<0.05); alveolar bone significantly increased in the experimental group (P<0.05). CONCLUSIONS: The clinical effect of PRF combined with Bio-oss on Class II furcation involvement are remarkable.

Escopoletin treatment induces apoptosis and arrests cell cycle at G0/G1 phase in the oral squamous cancer cell lines.

This study was aimed to investigate the antiproliferative effect of escopoletin on CAL 27 and CAL 33 oral squamous cancer cell lines. MTT assay and flow cytometry were used for the analysis of escopoletin effect on cell viability and apoptosis, respectively. Western blot analysis was used for the examination of cyclin and cyclin-dependent kinase expression after treatment of CAL 33 cells with escopoletin. The results showed a marked decrease in the viability of CAL 27 and CAL 33 cell lines after 48 h of escopoletin treatment. Treatment of CAL 33 cells with escopoletin led to the induction of apoptosis and arrest of cell cycle at G0/G1 phase. In the cells treated with escopoletin cyclin D1 and E expression was reduced and CDK1 expression was inactivated. The above findings suggest that escopoletin exhibits inhibitory effect on the oral squamous cancer through induction of apoptosis and arrest of cell cycle. Therefore escopoletin can be a promising candidate for the prevention of oral squamous cancer.

[Effect of Naringin on proliferation and osteogenic differentiation of bone marrow stromal cells in vitro].

PURPOSE: To investigate the differentiation and osteogenic activity of Naringin-induced bone marrow stromal cells (BMSCs) in dogs. METHODS: BMSCs were separated and cultured in vitro and identified by FCM. Then different concentration of Naringin (1×10⁻5, 1×10⁻6, 1×10⁻7, 1×10⁻8 and 1×10⁻9 mol/L) were added to cell culture media to induce BMSCs. The effect of Naringin on BMSCs was evaluated respectively by CCK-8 method and measuring the activity of alkaline phosphatase (ALP). The formation of nodules of calcium was detected by von Kossa staining. The data was analyzed with SPSS20.0 software package. RESULTS: The result of cell-surface marker displayed that the expression of CD34 and CD45 were negative while the expression of CD90 was positive. The values were 0.126%, 0.075% and 95.4%, respectively. Naringin could obviously promote cell proliferation, which exhibited the best effect on proliferation and osteogenic differentiation at concentration of 10-6 mol/L. Calcium nodule (von Kossa) staining was positive. CONCLUSIONS: Naringin-induced bone marrow stromal cells can be differentiated into osteoblasts. Naringin at the concentration of 10-6 mol/L can enhance the proliferation and osteogenic differentiation of BMSCs.

[Clinical application of modified casting post with zirconia crown in restoration of anterior teeth].

PURPOSE: To evaluate the clinical effect of modified casting post with zirconia crown in restoration of anterior teeth. METHODS: A total of 45 anterior teeth (32 patients) were treated with modified gold-alloy post-cores with zirconia crowns. All roots were greatly tilted to the labial side. Marginal integrity, anatomic form, surface, color and the gingival situation after restorations were assessed at 6 and 12 months with the California Dental Association (CDA) quality assessment system. RESULTS: The clinical outcome of 44 zirconia crowns achieved rank A (97.8%), while the gingival situation in one patient (2.2%) reached rank B. CONCLUSIONS: Modified gold-alloy post-core with zirconia crown contributes to the aesthetic effect of the anterior teeth which need to change the angle.

MiR-25-3p attenuates the proliferation of tongue squamous cell carcinoma cell line Tca8113.

OBJECTIVE: To investigate the effects of miR-25-3p on the occurrence, development and proliferation of tongue squamous cell carcinoma cells. METHODS: To establish tongue squamous cell carcinoma cell line Tca8113 that stably and highly express miR-25-3p using recombinant retroviral vector-mediated gene transfer method. The proliferation of transfected Tca8113 was detected by thiazolyl blue tetrazolium bromide (MTT) and cell colony formation assays. cyclinD1, p21(cip1) and p27(kip1) mRNA expressions in the transfected Tca-8113 were detected by quantitative PCR. cyclinD1, p21, p27(kip1), AKT, p-AKT, FOXO1 and p-FOXO1 expressions in the transfected Tca8113 were detected by western blot analysis. In addition, miR-25-3p expression in the tongue squamous cell carcinoma cell line and tissue specimen was also detected by quantitative PCR. RESULTS: Quantitative PCR showed that miR-25-3p expression in the tongue squamous cell carcinoma cell lines and tissue specimen was significantly lower than that in the adjacent tissue. MTT and cell colony formation assays showed that after miR-25-3p overexpression, the proliferation of transfected Tca8113 was obviously attenuated. Western blot analysis and quantitative PCR showed that after miR-25-3p overexpression, p21(cip1) and p27(kip1) expressions were upregulated, while cyclinD1, AKT, FOXO1 expressions were downregulated, and AKT and FOXO1 phosphorylation was inactivated in the transfected Tca8113 cells. CONCLUSIONS: MiR-25-3p inhibited the proliferation of tongue squamous cell carcinoma cells and regulated cell cycle-related protein expression, playing an important role in the occurrence and development of squamous cell carcinoma of the tongue.

[Establishment and experimental study of alveolar preservation before dental implantation in Beagle dogs].

PURPOSE: To establish the model of alveolar ridge preservation after tooth extraction for dental implant replacement, and to observe the effect of tissue engineered bone on osseointegration. METHODS: Isolated BMSCs were expanded and osteogenically induced in vitro. The tissue engineering complex was constructed with BMSCs/A-PCPC in vitro. Six extraction sockets, with three on each side, were created in the mandibles of four Beagle dogs by extracting the second, third and fourth premolars. BMSCs/A-PCPC were placed on one side of the extraction sockets, while autogenous bone, A-PCPC and nothing were placed on the other side as control. X-ray and CT scans were conducted 1day, 4 and 12 weeks after operation to detect the change of the alveolar ridge. The bone of sockets were harvested at 8-week post-implantation and subject to histological for evaluating. SPSS17.0 software package was used for data analysis. RESULTS: Radiographs demonstrated higher radiodensity in group of complex than in simple materials group, autogenous bone group after 4 weeks. Hard tissue biopsy at 12-week showed that bone activity of BMSCs/A-PCPC complex was better than the other groups. Spiral CT analysis showed that alveolar ridge of each group experienced a certain degree of absorption. At 12-week, the alveolar ridge height reduction values in A-PCPC group was smaller than in A-PCPC group, autogenous bone group and blank group (P<0.05). CONCLUSION: The BMSCs/A-PCPC complex is favorable for preservation of alveolar ridge.

[The accuracy of linear measurements of jaw implant regions obtained by multi-slice spiral CT and dental software].

PURPOSE: To evaluate the measurement accuracy by multi-slice spiral CT(MSCT) and dental software as a tool for preoperative design before tooth implantations. METHODS: Twenty volunteers with marker template underwent MSCT. The diameter of the front teeth crowns in the bucco-lingual direction were measured by MSCT using a standard dental software package. Postoperatively, the same distances were clinically measured using a sliding caliper. The consistency between the two measurements was analyzed by paired t test using SPSS17.0 software package. RESULTS: The difference of measurements was -0.04-0.21mm. There was no significant difference between the CT and the manual measurement of the crown diameter in the bucco-lingual direction(P>0.05). CONCLUSIONS: The results demonstrate that MSCT and marker template promises to be a valuable method for the measurement of jaw implant regions.

[Construction and detection of gene expression vector of the Von Hippel-Lindau in bone marrow stromal cells].

PURPOSE: To construct siRNA-VHL expression vector and detect the effect of VHL gene interference on BMSCs. METHODS: According to the dog's VHL gene sequences, four pairs of siRNA oligo were designed and synthesized. Using vector cloning kit reorganization, four pairs of double-stranded siRNA were inserted into the expression vector (pcDNA™ 6.2-GW/EmGFPmiR) and 4 siRNA expression plasmids (SR144-1,SR144-2,SR144-3,SR144-4) were constructed. With the vector universal primers, colony PCR was screened. The positive clones were sequenced to verify whether the sequence of insert fragments in recombinant clones was consistent with oligo sequences designed or not. Interference vector transiently transfected the BMSCs. qPCR and Western blot were used to detect the gene silencing effect. In order to improve transfection efficiency of siRNA-VHL as well as the effect of the VHL gene silencing, pLenti-mi-VHL was constructed. RESULTS: Through sequencing the plasmids cloned, the fragment sequences inserted in recombinant clones were consistent with the designed oligo sequences. After 24 h and 48 h transfection of BMSCs cells by plasmids, SR144-4 showed the best effect of interference by qPCR and Western blot. Through comparing the sequencing results, the inserted fragment sequences were completely correct and the pLenti-mi-VHL was successfully constructed. CONCLUSION: The siRNA-VHL expression vector for BMSCs is successfully constructed and applicable for further experiments. Supported by Research Fund of Science and Technology Commission of Shanghai Municipality (Grant No.9411954800) and Foundation for Open Project from Shanghai Key Laboratory of Stomatology (Grant No.S30206).

[The experimental study on porous calcium phosphate cement with bone marrow stromal cells for bone tissue engineering].

OBJECTIVE: To observe the biocompatibility of new biomaterials porous calcium phosphate (CPC) and ectopic bone formation of CPC with bone marrow stromal cells (BMSCs). METHODS: The BMSCs were cultured from Beagle dog and combined with the porous CPC with the best concentration after transfect green fluorescent protein (GFP). The adhesion and growth of BMSCs on CPC were observed under inversion, fluorescence and scanning electron microscopy. The ectopic bone formation were observed at the 8th week after CPC and BMSCs were implanted subcutaneously into nude mice. RESULTS: When BMSCs with CPC were cultured at the 1st day, cells were climbing out from CPC with normal morphology. At the 7th day cells can be seen protruding pseudopods, secretion of matrix. Bone formation could be seen histomorphologically at the 8th week. CONCLUSION: Porous CPC has good biocompatibility and is an ideal scaffold material for bone tissue engineering.

[Experimental study of subcutaneous osteogenesis by bone marrow stromal cells with A-PCPC in Beagle dogs].

PURPOSE: To study the osteogenic capability of the construct combined dog's bone stromal cells with active porous calcium phosphate cement (A-PCPC) in nude mice in vivo. METHODS: Isolated bone marrow stromal cells (BMSCs) were expanded and osteogenically induced in vitro. Their osteogenic phenotype was evaluated by cytochemistry. The tissue engineering complex was constructed with BMSCs/A-PCPC in vitro. After SEM scanning, the complex of BMSCs/A-PCPC was implanted into the subcutaneous tissue of the nude mice as experimental group, and A-PCPC as control group. The engineered bone was harvested 2,4,8weeks post-implantation and processed for HE staining, then evaluated by histology and histomorphometry. RESULTS: Cytochemistry showed alkaline phosphate activity, Von Kossa staining proved the formation of mineralization nodules. Scanning electron microscopy showed the cells adhered to the inner surface of the A-PCPC. HE staining showed a small group of woven bone formation 2 weeks later in the experimental group, while the formation of bone less in the control group. Woven bone turned into trabecular bone gradually at 4 weeks in the experimental group, while the control group showed a large number of bone-like tissue. Histomorphometry showed more mature bone in the experimental group than the control group at 8 weeks. CONCLUSIONS: The A-PCPC/BMSCs composites show good osteogenetic activity and could promote mineralization of the immature bone. It can be used as the bone tissue engineering scaffolds. Supported by Innovation Fund for Science and Technology Development of Pudong New District(Grant No. PKJ2009-Y19).

Can baseline magnetic resonance angiography (MRA) status become a foremost factor in selecting optimal acute stroke patients for recombinant tissue plasminogen activator (rt-PA) thrombolysis beyond 3 hours?

OBJECTIVE: We investigated whether baseline vessel status evaluated by magnetic resonance angiography (MRA) can be the foremost factor to classify acute ischemic stroke patients into subgroups for thrombolytic therapy within 3-6 hours of symptom onset. METHODS: Acute ischemic stroke patients beyond 3 hours after symptom onset were examined by stroke magnetic resonance imaging (MRI) (diffusion- and perfusion-weighted imaging, and MRA) before and after thrombolysis treatment within 24-48 hours. Stroke MRI was used to classify acute ischemic stroke patients into subgroups and select optimal patients for thrombolytic treatment. Clinical scores were compared to determine whether there were significant differences among subgroups. RESULTS: The difference in day 90 modified Rankin scale (mRS) between treated salvageable and untreated salvageable patients with recombinant tissue plasminogen activator (rt-PA) was remarkably statistically significant (p=0.02). Treated salvageable patients had more favorable clinical outcomes as compared with the untreated salvageable patients. Patients who did not have baseline artery occlusion were associated with more favorable clinical outcomes than untreated salvageable patients (p<0.001). The difference between treated salvageable and patients without artery occlusion in 90 day mRS score was not statistically significant (p=0.058). CONCLUSION: Baseline vessel status evaluated by MRA may be used as the first factor ahead of mismatch to categorize acute ischemic stroke patients into subgroups. Patients who do not have initial vessel occlusion may not need thrombolytic therapy.

[Stress distribution of two kinds of fixed prosthese supported by four or six dental implants in edentulous mandible].

PURPOSE: To investigate the stress distribution of 2 kinds of distal cantilevered fixed prostheses supported by 4 or 6 dental implants in edentulous mandible with three-dimensional finite element method. METHODS: The first design was 4 implants supported fixed prostheses with 6mm distal cantilever; the second design was 6 implants supported fixed prostheses with 6mm distal cantilever. Vertical force of 100 N was applied to the distal cantilever of fixed prostheses. RESULTS: The bone stress was concentrated at the cortex of mandible. The maximum bone stress was at the cortex near the second implants from the point that the vertical force was applied. The maximum implant stress was at the terminal and medium implant-prostheses interface. CONCLUSIONS: The bone around the terminal implant is easily to absorb and the terminal and medium implants are easily to fracture. The maximum stress of the implants and prostheses of two kinds of design are almost same(239Mpa). Two kinds of design can be applied in the clinic. 6 mm distal cantilevered fixed prostheses supported by 6 implants have the less bone stress than 4 mm distal cantilevered fixed prostheses in edentulous mandible.