Objective: To analyze the four intron SNPs of ARHGAP29 from previously reported GWAS results by bioinformatics, and performing information annotation and function prediction of target SNPs. Methods: Four statistically significant SNP loci were selected from the case-control experiment. The biological information of SNPs and the functions of related LncRNAs were predicted by using SNP database in NCBI website, 3DSNP database, RegulomeDB database, and LncSNP2.0 database. Results: Minor allele frequency (MAF) of rs72964308 was lower than 0.05, be excluded in subsequent analysis. The analysis of chromatin state showed that SNPs mainly locate in the enhancer region in various cell types, and also in the transcription initiation sites of rare tissues. The high DNA accessibility of the transcription factors that interacted with rs17396055 indicates a strong transcriptional activity. The prediction of LncRNA related with rs17396055 and rs7540684 showed that the disease with the highest correlation was cleft lip and palate. Conclusions: Rs3814019, rs17396055 and rs7540684 play regulatory roles in various cell types, and the LncRNA related to rs17396055 and rs7540684 may be associated with craniofacial development.

Objective: This study aimed to validate whether machine learning based on craniofacial-specific oral epithelium enhancers could be used to non-coding variants associated with orofacial cleft. Methods: Anti H3K27Ac ChIP-seq was performed in human immortalized oral epithelial cells (HIOEC), which were integrated published craniofacial super-enhancers. The sequences of these enhancers were used as training set for gapped k-mer SVM (gkm SVM) machine learning, which could summarize all the DNA sequence features. Delta SVM scores were employed to validate the effects of non-coding variants near IRF6, and dual luciferase assays were used for biological validation. Results: Published orofacial cleft-associated SNPs were more enriched in oral epithelium-specific enhancers. gkm SVM showed 350dupA mutation could significantly decrease enhancer activity, which was then validated using dual luciferase assays in HIOEC cells. Conclusions: The classifiers based on oral epithelium-specific enhancers are useful in nominating functional SNPs identified in orofacial clefting genome wide association studies.

Objective：To construct the Msx1-Cre mouse model. Methods：Design two sgRNA in the CRISPR/Cas9 system that target the Msx1 gene and construct homologous recombination fragments containing Cre sequences; Construct sgRNA vector plasmids and verify the target efficiency and off-target situation of sgRNA at the cellular level; Obtain F0 generation mice knocked into Cre sequence of Msx1 gene by microinjection method; Identification and functional verification of mouse models. Results：The designed sgRNA effectively cut the target site downstream of the Msx1 promoter in a targeted manner; We successfully obtained the Msx1Cre mouse model by microinjection; GFP was specifically located on the membrane of Msx1+ cells in Msx1Cre;R26RmTmG mice. Conclusions：The Msx1Cre mouse model was successfully constructed based on the CRISPR/Cas9 system.

Objective:?To investigate the effects of endothelin receptor A（ETRA）on osteogenic differentiation function of periodontal ligament stem cells (PDLSCs). Methods:?The small interfering RNA (siRNA) technique was employed to inhibit ETRA expression in PDLSCs and cells were cultured with ET-1 (100?nmol/L) for 14 days. The osteoblastic differentiation function of PDLSCs was observed by alkaline phosphatase staining. The expression of RUNX2 and OCN mRNAs and proteins in PDLSCs were detected by real-time RT-PCR and Western Blot, respectively. Results:?ETRA interference vector can effectively inhibit the expression of ETRA in PDLSCs. ET-1 significantly enhanced the expression of RUNX2 and OCN of PDLSCs（P＜0.05). However, ET-1 had no effect on the expression of RUNX2 and OCN of PDLSCs stably transfected by siETRA（P＞0.05）. Conclusions: These results indicate that endothelin-1 may stimulate the osteogenic differentiation of PDLSCs via ETRA.

Objective: To investigate the effect of titanium nanotubes formed by electrochemical anodization, loaded with genipin crosslinked chitosan hydrogel on biological activity of mouse bone marrow mesenchymal stem cells (BMSC) in vitro. Methods:?A two-step electrochemical anodization technique was used to prepare a regular array of titanium dioxide nanotubes on the substrate of titanium, chitosan coating was loaded by negative pressure method, and crosslinked with genipin. Field emission scanning electron microscope was used to observe the morphology of different substrates, and the contact angle measuring instrument was used to measure the hydrophilicity. The samples were co-cultured with BMSC, and cell adhesion and proliferation were measured by the CCK-8. Results:?Anodic oxidation method and negative pressure method was used to construct composite nano-structures. Field emission scanning electron microscopy showed particles on the surface, which showed increased hydrophilicity. The results of cell experiments confirmed that the nanocomposite had no obvious effect on the adhesion of BMSC, but promoted cell proliferation. Conclusions: Nanocomposites of titanium dioxide nanotubes loaded with genipin crosslinked chitosan hydrogel can improve BMSC activity.

Objective:?To investigate the effect of HSF1 in cancer-associated fibroblast (CAFs) on the growth and metastasis of subcutaneous xenotransplanted tumor of oral squamous cell carcinoma (OSCC) cell in nude mice. Methods:?Fibroblasts of OSCC and adjacent normal tissues were identified by Western-blot to obtain CAFs. CAFs-shHSF1 cells and control CAFs-shNC cells were constructed by lentivirus knockdown HSF1 in CAFs. The tumor cells Cal27 were mixed respectively with CAFs-shHSF1 cells and CAFs-shNC cells at 1:1 and inoculated into the right axilla of nude mice, and the formation and growth of the xenografted tumor were observed. The difference of tumor volume and weight between the two groups was statistically analyzed. HE staining and immunohistochemical staining were used to verify the metastasis and epithelial-mesenchymal transformation (EMT) of the xenografted tumor between the two groups. Results:?The protein abundance of α-SMA, FAP and FSP-1, the markers of CAFs, was significantly higher than those of normal fibroblasts. The tumor volume in the experimental group was significantly lower than that in the control group (P＜0.05), and there was no significant difference in body weight between the two groups (P＞0.05). Meanwhile, there was liver metastasis in both the experimental group and the control group, and the number of liver metastasis in the experimental group was less than that in the control group (P＜0.05).. Immunohistochemical staining data showed that α-SMA and Vimentin staining decreased and E-cadherin did not change significantly in Cal27+CAFs-shHSF1 group. Conclusions:?Knockdown of HSF1 in CAFs can significantly inhibit the growth and metastasis of OSCC in nude mice, which may play a role through partial EMT mechanism.

Objective: To investigate maxillary changes of rapid and slow maxillary expansion protocols through cone beam CT （CBCT）analysis. Methods: Sixteen patients (mean age 9.5years) were randomly distributed into 2 groups: rapid maxillary expansion (n = 8) and slow maxillary expansion (n = 8) via Hass appliance. Maxillary changes were measured by cone beam computer tomography analysis. The buccal bone plate thickness, palatal cleft width， nasal cavity width, and maxillary width were also recorded accordingly. Results: We found no statistical difference between the two groups in terms of palatal cleft width， nasal cavity width, and maxillary width（P＞0.05).Buccal bone plate thickness reduction was smaller in slow maxillary expansion group than those in rapid maxillary expansion group, althrough there was no statistical difference. Conclusions: Rapid and slow maxillary expansion could expand the maxillary width, and the slow maxillary expansion might result in light and continuous force which might be more favorable to periodontal tissue.

Objective: Changes of the position and shape of the condyle of TMJ before and after orthodontic-orthognathic treatment were measured to evaluate its influence on TMJ and to provide reference for the prediction of preoperative comprehensive therapeutic schedule.Methods:20 patients with severe skeletal Class III malocclusion were selected. CBCT of these patients were collected before orthodontic treatment, after decompensation and after orthodontic-orthognathic treatment. The diameter, gap and angle of TMJ were measured through CBCT and compared the changes before and after treatment. Results: There was no statistical difference in the measurement items of TMJ between the left and right sides (P > 0.05) before and after treatment. There was a significant difference (P < 0.05) in the horizontal angle of the left condyle before and after decompensation of orthodontic treatment, while there was no significant difference (P > 0.05) in the other three-dimensional measurement items of TMJ. The horizontal angle of the left and right condyles, the supra articular space and the anterior articular space increased significantly before and after treatment (P < 0.05). Conclusions: The preoperative orthodontic treatment had no significant effect on the temporomandibular joint. But after orthodontic-orthognathic surgery, the condyle position changed backward and downward.

The oral biomedical laboratory is an important place for to carry out different research of stomatology and cultivate high-quality talents. Here we summarize the potential safety hazard and management issues in oral biomedical laboratories. We also put forward some strategies to reduce the risks and improve the level of safety protection.