Effects of p38 MAPK signal pathway inhibition on upper lip scar hyperplasia at different time in rabbits / 中华医学美学美容杂志

Objective To investigate the effect of p38MAPK gene silencing recombinant adeno-virus on the expression of target gene in different time and to detect the effect of p 38MAPK signal pathway on the upper lip scar hyperplasia at different time to determine the optimal scar treatment time .Methods The adenovirus vector was injected into the scar tissue in 0 week ,1 week and 2 week after cheiloplasty in rabbit .The specimens were harvested in 3 week postoperatively .Four methods in-cluding Sirius red staining ,immunohistochemical staining (IHC) ,Western blotting (WB) ,real-time PC (RT-PCR) were used to quantitatively and quantitatively detect the relative expression levels of p38MAPK and scar-related factors (col Ⅰ ,col Ⅲ ,MM P1 ,TIMP1) .Results Sirius red staining and immunohistochemical staining showed that in 1st week the expression of col Ⅲ and MMP1 in scar tis-sue was significantly higher than that in 0 week and 2 week after operation and the expression of col Ⅰand TIMP1 was significantly less than that in 0 week and 2 week after operation .The results of WB and RT-PCR were consistent with that of IHC .Conclusions After injection into the upper lip scar tis-sue with adenovirus in 1 week ,the degree of scar hyperplasia is the least .

[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.

[Influence of 7-dehydrocholesterol reductase gene silencing on the fusion of mouse palatal shelves].

OBJECTIVE: RNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves. METHODS: The pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV- siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n = 10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n = 10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experimental group (n = 10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses. RESULTS: SEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves. RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a significant difference (P < 0.05). CONCLUSION: Results indicate that Dhcr7 gene silencing affects the fusion of palatal shelves. Thus, Dhcr7 gene may serve a function in the normal development of palates.

Influence of 7-dehydrocholesterol reductase gene silencing on the fusion of mouse palatal shelves / 华西口腔医学杂志

<p><b>OBJECTIVE</b>RNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves.</p><p><b>METHODS</b>The pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV- siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n = 10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n = 10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experimental group (n = 10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses.</p><p><b>RESULTS</b>SEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves. RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a significant difference (P < 0.05).</p><p><b>CONCLUSION</b>Results indicate that Dhcr7 gene silencing affects the fusion of palatal shelves. Thus, Dhcr7 gene may serve a function in the normal development of palates.</p>

7-Dehydrocholesterol reductase regulated the palatal development by the sonic hedgehog-bone morphogenetic protein 2 signal pathway / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of 7- dehydrocholesterol reductas (Dhcr-7) gene silencing on the palatal development by sonic hedgehog (Shh)-bone morphogenetic protein2(BMP-2) signal pathway in vitro.</p><p><b>METHODS</b>A total of 60 pairs of palatal shelves fromgestation day (GD) 13.5 mouse embryos were divided into three groups (A, B, C) of 20 randomly. In group A (control), palatal shelves were cultured with medium containing no cholesterol.In group B (Dhcr-7-siRNA), palatal shelves were cultured without cholesterol medium but containing Dhcr-7 siRNA adenovirus. After 48h, the culture medium of groups A and B were changed with medium without cholesterol. In group C (cholesterol), palatal shelves were cultured without cholesterol medium but containing Dhcr-7 siRNA adenovirus. After 48h, the culture medium of group C was changed with medium containing 600 mg/L cholesterol. After 72h again, tissues dyeing and scanning electron microscope (SEM) technique were used to observe morphological changes of palates. Both RT-PCR and Western blottingtechniques were used to measure mRNA and protein expressions for Dhcr-7, Shh, and BMP-2, respectively.</p><p><b>RESULTS</b>The tissues dyeing and SEM showedthat the palates fusedin groups A and Candthe palates did not fuse in group B eventually. The expression of both mRNA and proteins for Shh and BMP-2 in group B wasdecreased with the Dhcr-7 reduction. In group B, the mRNA and protein expression of Shh was separately 0.063±0.018 and 0.092±0.065;the mRNA and protein expression quantity of BMP- 2 was separately 0.054±0.018 and 0.049±0.021. In group A, the mRNA and protein expression of Shh was separately 0.667±0.093 and 0.639±0.078;the mRNA and protein expression of BMP-2 was separately 0.591 ± 0.043 and 0.569 ± 0.081. The difference of Shh and BMP- 2 mRNA and protein expression between A and B group were statistically significant separately (P < 0.05). The expression of both mRNA and protein for Dhcr-7 (0.074±0.034 and 0.075±0.028) did not changebasicallyin group C, compared with the Dhcr- 7expression of mRNA and protein (0.083±0.045; 0.067±0.065) in group B, the difference wasnot statistically significant(P > 0.05). In group C, the mRNA and protein expressionof Shh (0.649±0.085 and 0.608±0.092) and BMP-2 (0.578±0.062 and 0.548±0.065) were significantly increased. The difference of Shh and BMP-2 mRNA and protein expression between B and C group were statistically significant separately (P < 0.05).</p><p><b>CONCLUSIONS</b>Dhcr-7 could influence the expression of Shh and BMP-2. Dhcr-7 reductase regulated the palatal development by the Shh-BMP-2 signal pathway.</p>

Evaluation of osteogenesis of alveolar bone graft by cone beam CT / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the osteogenesis of alveolar bone graft (ABG) in patients with alveolar cleft by cone beam CT (CBCT).</p><p><b>METHODS</b>ABG surgery was performed in 20 patients with unilateral complete alveolar cleft. The patients were followed up for 3 and 6 months after surgery and the osteogenesis of the bone graft was evaluated by CBCT. The bone density and the height of labial and palatal bone graft area were analyzed.</p><p><b>RESULTS</b>There was no significant difference in the bone density between 3 months [(403.79 ± 64.70) HU] and 6 months[(411.45 ± 42.62) HU ] (P = 0.329).However, there was significant difference in bone height in the labial and palatal side between 3 months and 6 months (labial P = 0.020, palatal P = 0.008).</p><p><b>CONCLUSIONS</b>The osteogenesis was the best 3 months after bone graft. The following treatment can start in this stage.</p>

Recombinant bovine basic fibroblast growth factor gel prevents dry socket syndrome after toothextraction / 中国组织工程研究

BACKGROUND:Recombinant bovine basic fibroblast growth factor is a manifold effect cytokine which can promote angiogenesis, wound healing, tissue repair and bone regeneration. Recombinant bovine basic fibroblast growth factor with good histocompatibility is easy to operate and has been widely used in oral and maxil ary surgery. OBJECTIVE:To evaluate the effect of recombinant bovine basic fibroblast growth factor against dry socket syndrome after tooth extraction. METHODS:A total of 160 patients who had been extracted mandibular third molar were selected and randomly divided into two groups. In the experimental group, recombinant bovine basic fibroblast growth factor was put into the sockets after mandibular third molars were extracted, while in the control group, we let the wounds to be healed natural y without any materials. The incidence of dry socket syndrome was observed and compared between two groups at 3 days, 5 days and 1 week after tooth extraction. RESULTS AND CONCLUSION:One patient had dry socket after operation in the experimental group, and the incidence was 1.25%. In the control group, 10 patients suffered from dry socket, and the incidence was 12.5%. There was a significant difference in the incidence of dry socket between the two groups (P<0.01). There was visible granulation tissue within the tooth socket after tooth extraction in the experimental group, and extraction sockets narrowed and were fil ed with granulation tissues, which was 1-2 days earlier than the control group. No al ergies, tissue hyperplasia and other local and systemic reactions occurred in patients receiving implantation of recombinant bovine basic fibroblast growth factor gel. These findings indicate that local implantation of recombinant bovine basic fibroblast growth factor gel after mandibular tooth extractions can speed up the healing of dental extraction wounds.

Oral cancer cells with different potential of lymphatic metastasis displayed distinct biologic behaviors and gene expression profiles.

OBJECTIVE: Oral squamous cell carcinoma (OSCC) often spreads from the primary tumor to regional lymph nodes in the early stage. Better understanding of the biology of lymphatic spread of oral cancer cells is important for improving the survival rate of cancer patients. METHODS: We established the cell line LNMTca8113 by repeated injections in foot pads of nude mice, which had a much higher lymphatic metastasis rate than its parental cell line Tca8113. Then, we compared the biologic behaviors of cancer cells between them. Moreover, microarray-based expression profiles between them were also compared, and a panel of differential genes was validated using real-time-PCR. RESULTS: In contrast to Tca8113 cells, LNMTca8113 cells were more proliferative and resistant to apoptosis in the absence of serum, and had enhanced ability of inducing capillary-like structures. Moreover, microarray-based expression profiles between them identified 1341 genes involved in cell cycle, cell adhesion, lymphangiogenesis, regulation of apoptosis, and so on. Some genes dedicating to the metastatic potential, including JAM2, TNC, CTSC, LAMB1, VEGFC, HAPLN1, ACPP, GDF9 and FGF11, were upregulated in LNMTca8113 cells. CONCLUSION: These results suggested that LNMTca8113 and Tca8113 cells were proper models for lymphatic metastasis study because there were differences in biologic behaviors and metastasis-related genes between them. Additionally, the differentially expressed gene profiles in cancer progression may be helpful in exploring therapeutic targets and provide the foundation for further functional validation of these specific candidate genes for OSCC.

Altered phenotype of lymphatic endothelial cells induced by highly metastatic OTSCC cells contributed to the lymphatic metastasis of OTSCC cells.

The lymphatic endothelial cell (LEC) is an interactive surface for cancer cells. This article aims to explore cancer cell-induced changes of LEC, and study the tumor-lymphatic endothelium interaction. Here, LECs were co-cultured with highly and poorly metastatic tongue cancer cells. The differences in biologic behaviors and gene expression profiles between them were examined. The results showed that LECs induced by highly metastatic cancer cells displayed abnormal biologic behaviors, and could secrete chemokines to promote the migration of cancer cells. Therefore, biologic properties and functional status of LECs in oral tongue squamous cell carcinoma (OTSCC) might be a positive factor in lymphatic dissemination.

An experimental study of folic acid rivaling methylenetetrahydrofolate reductase (MTHFR) gene / 实用口腔医学杂志

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

A histomorphologic and enzyme histochemical study of masticatory muscles affected by distraction osteogenesis of mandible / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The purpose of this study was to study the masticatory muscles affected by distraction osteogenesis of the mandible.</p><p><b>METHODS</b>The distraction osteogenesis (DO) was applied to distract the left mandible of 6 mongrel dogs that were divided into three experimental groups. After different distraction phase and consolidation phase, the masseter and the digastric muscle were taken out. The specimens were stained using hematoxylin/eosin and enzyme histochemistry. Afterwards, the specimens were observed with a light microscope to study the morphologic changes of the muscles. The contents of enzyme in the different groups were measured by VIDAS.</p><p><b>RESULTS</b>The masseter showed consequently atrophy, but the digastric muscle showed a progress of histomorphologic reconstruction, including atrophy and hypertrophy. The changes of the contents of enzyme and histomorphology were identical in the masticatory muscles.</p><p><b>CONCLUSION</b>The digastric muscle parallel to the vector of mandibular distraction adapts the distraction by the way of atrophy, regeneration and hypertrophy. And the contents of enzyme appear to decrease at the beginning, increase afterwards, and return to the normal level finally. But the masseter perpendicular to the vector of mandibular distraction shows consequent atrophy, and the contents of enzyme consequently decrease, which means the metabolism decrease.</p>