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BACKGROUND:In consideration of the food safety and ecological safety of transgenic plants,the retention of marker genes is the primary safety issue affecting transgenic plants. OBJECTIVE:Based on the principle of immune caries prevention,our research team successfully constructed the plant anti-caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 for these two caries causing virulence factors surface protein and glucosyltransferase,which provides a basis for the research and development of transgenic plant vaccine. METHODS:In this study,the selective marker genes Km and GUS in the plant caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 were removed by DNA recombination technology through a series of steps such as DNA fragment separation,connection,transformation,clone detection,and sequencing. RESULTS AND CONCLUSION:The efficiency of marker gene removal was 99%.This study has laid a good experimental foundation for the safe production of transgenic plant vaccine against dental caries,and also provided ideas for the construction of other plant vaccine vectors.
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BACKGROUND:The application of orthodontic force triggers autophagy in the periodontal tissue via diverse signaling pathways,augmenting or attenuating the activity of relevant cell types such as periodontal ligament cells,osteocytes,osteoclasts,and osteoblasts,thus facilitating the process of periodontal remodeling. OBJECTIVE:To review the research progress in orthodontic force mediated autophagy in periodontal tissue and its impact on orthodontic tooth movement. METHODS:The PubMed,Web of Science,China Biology Medicine disc and CNKI were searched for literature published from 2010 to 2023 to summarize the progress in orthodontics-related autophagy.And 76 papers were finally included in the analysis and discussion. RESULTS AND CONCLUSION:Orthodontic force can trigger a series of biochemical signal changes through periodontal mechanical receptors and aseptic inflammation they cause,leading to autophagy in periodontal tissue.Subsequently,autophagy generates corresponding feedback through cascaded amplified signaling pathways such as Phosphoinositide 3-kinase/protein kinase B,Hippo,and mitogen-activated protein kinase pathways,promoting periodontal tissue remodeling and ultimately achieving tooth movement and stability.Orthodontic force-induced autophagy can differentially regulate bone resorption on the tooth pressure side and bone formation on the tension side.Related targets have good prospects in the clinical application of orthodontic treatment.Orthodontics and autophagy have complex mechanisms.However,existing research has only focused on exploring the role of autophagy in orthodontic tooth movement.Further exploration is needed to investigate the mutual regulatory effects between autophagy and orthodontic tooth movement,as well as the interactions between upstream mechanical receptors and signaling pathways involved in related pathways.
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Objective To explore the clinical effect of procaterol hydrochloride combined with Xiaokechuan capsule in the treatment of cough variant asthma(CVA)and its impact on serological indicators,airway function of children.Methods A total of 124 children with CVA admitted to the Zigong First People's Hospital from March 2019 to April 2022 were selected as the research subjects.The children were divided into control group and observation group according to random number table method,with 62 cases in each group.The children in the control group were treated with procaterol hydrochloride,and the children in the observation group were treated with procaterol hydrochloride and Xiaokechuan capsule for two weeks.The clinical efficacy of children was compared between the two groups after treatment.The cough scores during the day and night of children were evaluated in the two groups before and 2 weeks after treatment.The serum high mobility group protein B1(HMGB1),Toll like receptor 4(TLR4),nuclear factor-κB(NF-κB),interleukin-4(IL-4),interferon-γ(INF-γ)levels of children in the two groups were measured by enzyme linked immunosorbent assay before and 2 weeks after treatment,and the ratio of INF-γ/IL-was calculated.The 25%maximal expiratory flow-volume(MEF25),50%maximal expiratory flow-volume(MEF50),75%maximal expiratory flow-volume(MEF75)of children in the two groups were measured by lung function detector before and 2 weeks after treatment.The adverse reactions of children in the two groups were recorded during treatment.Results The total effective rate of children in the control group and observation group was 82.26%(51/62)and 95.16%(59/62),respectively;the total effective rate of children in the observation group was significantly higher than that in the control group(P<0.05).There was no significant difference in cough scores during the day and night of children between the two groups before treatment(P>0.05);the cough scores during the day and night of children after treatment were significantly lower than those before treatment in the two groups(P<0.05);after treatment,the cough scores during the day and night of children in the observation group were significantly lower than those in the control group(P<0.05).There was no significant difference in serum HMGB1,TLR4,NF-κB levels of children between the two groups before treatment(P>0.05);the serum HMGB1,TLR4,NF-κB levels of children after treatment were significantly lower than those before treatment in the two groups(P<0.05);after treatment,the serum levels of HMGB1,TLR4,and NF-κB of children in the observation group were significantly lower than those in the control group(P<0.05).There was no significant difference in MEF25,MEF50,and MEF75 of children between the two groups before treatment(P>0.05);the MEF25,MEF50,and MEF75 of children after treatment were significantly higher than those before treatment in the two groups(P<0.05);after treatment,the MEF25,MEF50,and MEF75 of children in the observation group were significantly higher than those in the control group(P<0.05).There was no significant difference in serum IL-4,INF-γ levels and the ratio of INF-γ/IL-4 of children between the two groups before treatment(P>0.05);the serum IL-4 level of children after treatment were significantly lower than those before treatment,the INF-γ level and the ratio of INF-γ/IL-4 were significantly higher than those before treatment in the two groups(P<0.05);after treatment,the serum IL-4 level of children in the observation group was significantly lower than that in the control group,the INF-γ level and the ratio of INF-γ/IL-4 were significantly higher than those in the control group(P<0.05).All children had good drug tolerance during the treatment period,and no significant adverse drug reactions were observed.Conclusion The combination of Xiaokechuan capsules and procaterol hydrochloride has a significant therapeutic effect for pediatric CVA,and its mechanism of action may be related to the regulation of HMGB1-TLR4-NF-κB signal pathway.
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Oral submucous fibrosis (OSF) can cause various oral dysfunctions in patients and can turn into oral cancer. The causes and processes of OSF malignant transformation involve betel nut chewing, vascular atrophy, tissue hypoxia, cell cycle changes, aging, autophagy, and changes in cancer/cancer suppressor genes and microRNAs. It is of great significance to study the causes and process of OSF malignant transformation for the treatment and prevention of OSF malignant transformation.
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Forkhead box protein O1 (FOXO1) has been extensively studied as a tumor suppressor. In oral squamous cell carcinoma, studies have demonstrated that FOXO1 can inhibit tumor cell oxidative stress, stemness and epithelial-mesenchymal transition, and promote tumor cell autophagy and apoptosis. FOXO1 may serve as a potential target for the treatment of oral squamous cell carcinoma.
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Objective:To explore the influence of flipped classroom combined with case-based learning (CBL) teaching mode on teaching effect of X-ray cephalometric analysis in orthodontic experimental class.Methods:In the study, 70 undergraduates majoring in stomatology were selected as research objects and randomized into teaching reform group and traditional group in average. Flipped classroom combined with CBL teaching mode was adopted in teaching reform group, while traditional group adopted traditional teaching mode. Questionnaire was used to investigate the students' evaluation on the two teaching modes, and the accuracy of the measurement of X-ray cephalometrics of the two groups was compared and analyzed respectively. GraphPad Prism 7.03 software was used to perform t test and chi-square test. Results:Teaching reform group believed that this kind of teaching mode could improve the ability of self-study, communication coordination ability, independent problem solving ability, and logic and language expression ability, deepen the understanding of the project connotation, and apply the X-ray cephalometrics to the actual clinical case analysis and diagnosis. Compared with the traditional group, there were significant differences ( P<0.05). In the accuracy evaluation of measurement values, the absolute values of 70% measurement items in teaching reform group were higher than those in the traditional group, but the difference was not statistically significant ( P>0.05). Only in bone type Ⅲ malocclusion, the absolute values of Y-axis angle and FMA were lower than those of the traditional group, and the difference was statistically significant ( P<0.05). Conclusion:Flipped classroom combined with CBL teaching mode can help students improve self-study ability, communication cooperation ability and the ability of solving problem independently and clinical application. However, the accuracy evaluation of measurement values in teaching reform group were lower than that of the traditional group, indicating that this teaching method in the experimental teaching of X-ray cephalometric analysis needs to be further improved.
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Objective To study the influence of parental generation coal-burning-borne fluorosis on tooth development of their offspring.Methods High fluoride air model was established on the basis of burning coal habit of the epidemic areas.Fluoride feed was made of coal drying corn from the epidemic areas.Totally 48 SD rats were randomly divided into 4 groups with male to female ratio of 1:1 by random number table method.Rats in high,middle and low fluoride groups were put in the high fluoride air room and feed food with 40,25 and 10 mg/kg fluorine,and the control group was put in normal air room and feed normal food.After 8 weeks,rats were mating and parturition.Tooth eruption time of offspring rat was observed;and dental fluorosis incidence,the tooth length and fluorine content were observed at 21 d.Results In high and middle fluoride groups [(6.83 ± 0.94),(6.25 ± 1.06) d],tooth eruption time of offspring rat was later than that of control group [(5.34 ± 0.89) d,all P < 0.01].At 21 d,dental fluorosis was observed in the lower incisors of the high and middle fluorine groups;compared with control group [(5.21 ± 0.19) mm,(223.00 ± 14.08) μg/kg],the tooth length was decreased [(4.83 ± 0.22),(4.96 ± 0.25) mm,P < 0.01or < 0.05],and tooth fluoride content was increased [(362.64 ± 20.35),(289.79 ± 19.18) μg/kg,all P < 0.01].Dental fluorosis incidence of offspring rats was positively correlated with the fluorine dose (r =0.704,P < 0.01).Conclusion Parental generation rats’ intaking excessive fluoride can affect offspring rats tooth development and dental fluorosis,which is related to the fluorine dose.
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Objective To establish a fluorosis model induced by coal burning and in ameloblasts of rat offsprings.Methods High fluoride air model was established based on the burning coal habit of the epidemic areas.Fluoride feed was made of corn dried by coal burning.Thirty-six SD rats were divided into 3 groups by random number table method according to body weight in a male and female ratio of 2 ∶ 1∶ in the high fluoride air room and rats were feed food with fluorine at 40 mg/kg (high fluoride group),25 mg/kg (low fluoride group);in the normal air room and rats were feed food with fluorine at 0 mg/kg (the control group),12 rats in each group.Mating litter in a ratio of 2 ∶ 1 at the end of 8 weeks.The offsprings were killed on postnatal day 3 and 7 to make mandible sections.Specimens were prepared for light microscope examination to observe the morphological changes of ameloblasts in the tooth germ.Results At the end of 0,2,4,6 and 8 weeks,serum fluoride of the high fluoride group were (0.031 ± 0.003),(0.060 ± 0.006),(0.085 ± 0.006),(0.110 ± 0.007) and (0.134 ± 0.008) mg/L;serum fluoride of the low fluoride group were (0.031 ± 0.003),(0.046 ± 0.005),(0.077 ± 0.006),(0.091 ± 0.007) and (0.104 ± 0.007) mg/L;serum fluoride of the control group were (0.030± 0.003),(0.037 ± 0.002),(0.044 ± 0.002),(0.046 ± 0.003) and (0.049 ± 0.003) mg/L.At the end of 2,4,6 and 8 weeks,serum fluoride of high fluoride group and low fluoride group were significantly higher than that of control group (all P < 0.05).At 7 d,offspring rats in high fluoride group,adamantoblasts were in distortions and vacuole changes,but offspring rats in low fluoride group and the control group had no abnormality.Conclusion By providing rat with high fluoride air and food,we could establish a fluorosis model induced by coal burning in ameloblasts of rat offsprings.
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Endemic dental fluorosis has been reported in some regions of the world. China seemed to have high prevalence of endemic dental fluorosis, especially in southwest China. It is now most likely that excessive fluoride intake during enamel development play a key role in the pathogenesis of dental fluorosis. However, excessive intake of fluoride-induced cellular and molecular mechanisms of dental fluorosis are not entirely conclusive. Scholars at home and abroad have made a lot of research on pathogenesis of enamel fluorosis by using various experimental techniques. More recent studies mainly suggest that endoplasmic reticulum stress and calcium overload-associated apoptotic pathway may participate in fluoride excess-evoked pathogenesis of dental fluorosis. Furthermore, the functional changes of enamel matrix protein and protease activity may be involved in the pathological event. This paper summarized the recent research progress on this topic.
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BACKGROUND:Orthodontic treatment is a mechanical force for tooth to cause the remodeling of periodontal tissue, produced by the tooth movement. The main aspect of orthodontic periodontal tissue remodeling is the alveolar bone. Insulin-like growth factor is an important factor in the remodeling of periodontal tissue, which plays an important role in the growth, differentiation and growth of the cels. OBJECTIVE:To review the role of insulin-like growth factors in periodontal tissue remodeling. METHODS: A computer-based retrieval of PubMed, CNKI and Guizhou Province Digital Library Database was performed to search articles related to the role of insulin-like growth factors in periodontal tissue remodeling. The keywords were “insulin-like growth factor; periodontal tissue; remodeling” in English and Chinese, respectively. RESULTS AND CONCLUSION: Insulin like growth factor belongs to the insulin family, a kind of peptides, which can promote the migration, proliferation, differentiation, colagen and matrix synthesis and alkaline phosphatase activity of osteoblasts, fibroblasts and mesenchymal cels in the periodontal ligament. It also plays an important role in the repair of injury. During orthodontic treatment, the use of suitable orthodontic force combined with insulin-like growth factor can promote periodontal tissue remodeling, accelerate the orthodontic tooth movement and shorten the treatment time for patients.
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Objective SD rats were immuned with the transgenic tomatoes which carried fused gene of a region of PAc Strep-tococcus mutants and cholera toxin B subunit.The immunogenicity was tested to explore secure and economic edible vaccines a-gainst dental caries.Methods A total of 18 eighteen-day-old female SD rats were subdivided randomly into three groups:the exper-imental group which were fed with transgenic tomato juice containing chimaera protein PAcP/CTB;the positive control group which were treated with deactivated S.mutans;the negative control group which were not treated with transgenic tomato juice.Rats were immuned once per week for four weeks.Blood and saliva were collected at one day before the first immunity and one week after each immunization.IgG of blood serum and SIgA of saliva were detected using the enzyme-linked immunosorbent assay (indirect ELISA) testing.On day 70,rats were terminated.The maxillary and mandibular bones were subsequently taken out to count dental caries′scores.Results Post immunization,the experimental group and the positive control group had statistical significant levels of speci-ficity IgG in serum and SIgA in saliva compared to the negative control group (P <0.05).There was a significance difference be-tween the experimental group and the negative control group except in Dx levels of caries loss (P <0.05).Conclusion The targeted protein expressed on the transgenic tomatoes is immunogenic,which can effectively induce mucous membrane immune response and the systematical immunoreaction to suppress the occurrence of the dental caries.
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<p><b>OBJECTIVE</b>To investigate the synergistic effect of transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factor-BB (PDGF-BB) on the expression of integrin beta3, in periodontal membrane of rat orthodontic tooth.</p><p><b>METHODS</b>An orthodontic tooth movement model was established. Up to 32 experimental rats were randomly divided into four groups according to a random number table. The four groups were injected with 1% PBS, TGF-beta1 (5 ng), PDGF-BB (10 ng), and combined TGF-beta1 (5 ng) and PDGF-BB (10 ng) in the buccal submucosal, respectively. The volume injected in each group was 0.1 mL. The animals were then sacrificed on the 10th day. The left maxillary first molar and periodontal tissue were taken. Different expressions of integrin beta3 were detected in periodontal tissues through immunohistochemistry. Mean optical density (OD) values of the positive fields were examined. The data obtained were analyzed through ANOVA. The data followed normal distribution, and were compared via t-test.</p><p><b>RESULTS</b>Compared with the control groups, the expression of integrin beta3 was higher in the experimental groupin tension sides (P < 0.01). Significant differences in tension sides between the single-injection groups and the combined group were observed (P < 0.01). Compared with the control groups, the expression of integrin beta3 was higher in the experimental group in compression sides (P < 0.05). In addition, there was no significant differences in compression sides between the single-injection groups and the combined group (P > 0.05).</p><p><b>CONCLUSION</b>In terms of local regulatory factors, TGF-beta1 combined with PDGF-BB enhance the expression of integrin beta3 in the periodontal membrane and accelerate periodontal remodeling. The synergistic effect of the two growth factors is better than the single growth factor.</p>
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Animaux , Rats , Intégrine bêta3 , Molaire , Desmodonte , Facteur de croissance dérivé des plaquettes , Protéines proto-oncogènes c-sis , Mouvement dentaire , Facteur de croissance transformant bêta-1Résumé
Objective:To survey the expression of MMP-1,MMP-2 of human periodontal ligament cells(HPDLCs)treated by tea polyphenols(TP)and lipopolysaccharide(LPS).Methods:HPDLCs were in vitro cultured in vitro and treated by TP(200 μg/ml) and /or LPS(100 μg/ml)for 24,48 and 72 h respectively,the secretion of MMP-1 and MMP-2 were examined by ELISA,MMP-1 and MMP-2 mRNA expression was examined by real-time PCR.Results:The secression and mRNA expression of MMP-1 and MMP-2 of HPDLCs increased by LPS treatment and significantly inhibited by TP at the different times.Conclusion:TP can inhibit the col-lagen degradation of HPDLCs mediated by LPS.
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BACKGROUND:Orthodontic tooth movement is based on the periodontal tissue remodeling. In the exogenous factors accelerating orthodontic tooth movement, Chinese herbal medicine has become a research hotspot because of its wide resources, low cost, easy to extract, mild effect, smal toxic, less side effects and drug resistance. OBJECTIVE: To summarize the role of Chinese herbal medicine in the periodontal tissue remodeling during orthodontic tooth movement. METHODS:A computer-based retrieval of CNKI, Wanfang and PubMed databases was performed for articles related to Chinese herbal medicine for improving orthodontic tooth movement published before 2014. The keywords were “Chinese herbal medicine, orthodontic tooth movement, periodontal tissue remodeling” in Chinese and English, respectively. RESULTS AND CONCLUSION:Erigeron breviscapus, Salvia, teasel, Drynaria, baicalin, evening primrose oil as Chinese herbs are most widely used in the promotion of periodontal tissue remodeling, characterized as wide resources, low cost, easy to extract, mild effect, low toxicity, less drug resistance. In the clinical orthodontic treatment, it is hoped to accelerate orthodontic tooth movement and shorten the treatment time. Therefore, under the appropriate corrective force, Chinese herbs can be used properly to improve periodontal tissue repair and remodeling, which can improve the microcirculation of periodontal tissue, increase the local blood flow, promote bone formation and repress bone resorption.
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OBJECTIVE@#To over-express cyclin-dependent kinase 2-associated protein 1 (CDK2-AP1) gene, and investigate its effect on the proliferation and cell cycle regulation in breast cancer cell line MCF-7.@*METHODS@#CDK2-AP1 gene coding region was cloned into lentivirus vector. Lentivirus particles were infected into MCF-7 cells to upregulate the expression of CDK2-AP1 gene. The expression level of CDK2-AP1 was detected at both mRNA and protein levels by real-time PCR and Western blot. MTT assay, colony formatting assay, and flow cytometry were performed to detect the change of proliferation and cell cycle in MCF-7 cells. We examined the expression of cell cycle associated genes (CDK2, CDK4, P16Ink4A, and P21Cip1/Waf1) followed by CDK2-AP1 over-expression by Western blot.@*RESULTS@#CDK2-AP1 gene was up-regulated significantly at both mRNA (6.94 folds) and protein level. MTT based growth curve, colony formatting assay and flow cytometry showed that CDK2-AP1 over-expression lentivirus inhibited the proliferation of MCF-7 cells with statistical difference (P<0.05). In addition, with CDK2-AP1 over-expression, MCF-7 cells were arrested in G1 phase accompanied by apoptosis. Western blot showed that the expression level of P21Cip1/Waf1 and P16 Ink4A was upregulated, while the expression level of CDK2 and CDK4, members of the CDK family, was downregulated.@*CONCLUSION@#CDK2-AP1 gene plays a cancer suppressor role in breast cancer. Its function includes inhibiting the proliferation of MCF-7 cells and arresting the cell cycle in G1 phase.
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Humains , Tumeurs du sein , Cycle cellulaire , Division cellulaire , Prolifération cellulaire , Kinases cyclines-dépendantes , Régulation négative , Cellules MCF-7 , Protein kinases , Génétique , Métabolisme , Protéines suppresseurs de tumeurs , Génétique , MétabolismeRésumé
<p><b>OBJECTIVE</b>To investigate the combined effects of platelet-derived growth factor-BB (PDGF-BB) and insulin-like growth factor-I (IGF-I) on the expression of integrin (beta3 protein in the periodontal tissues of the compressing side of orthodontic tooth in rats.</p><p><b>METHODS</b>Establishing orthodontic movement model in Sprague-Dawley rats, which were injected with 10 ng PDGF-BB, 200 ng IGF-I alone or in combination in the buccal submucosal area of the orthodontic tooth. The injection was applied every other day. The experiment continued for ten days and then the rats were sacrificed. The expression of integrin (beta3 protein in periodontal ligament tissues of the compressing side was detected by immunohistochemical techniques.</p><p><b>RESULTS</b>The expression of integrin (beta3 protein in periodontal ligament tissues of the compressing side of each experimental group was significantly increased compared with that of the control group (P<0.01). Meanwhile the maximum mean optical density value of integrin (beta3 protein expression was attained in the combination group which showed a significant increase compared with the PDGF-BB group (P<0.05) and the IGF-I group (P<0.01).</p><p><b>CONCLUSION</b>The combination of exogenous PDGF-BB and IGF- I in orthodontic tooth movement may enhance the expression of integrin (beta3 protein in periodontal ligament cells and PDGF-BB and IGF-I may have a synergistic effect during orthodontic tooth movement.</p>
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Animaux , Rats , Cellules cultivées , Fibroblastes , Facteur de croissance IGF-I , Intégrines , Desmodonte , Facteur de croissance dérivé des plaquettes , Protéines proto-oncogènes c-sis , Rat Sprague-Dawley , Somatomédines , Mouvement dentaireRésumé
Objective:To observe the anticaries effects of the vaccine gene pcDNA3- PAc against dental caries in BALB/c mice by different routes. Methods: BALB/c mice were immunized with the recombinant plasmid pcDNA3- PAc by the submandibular gland- target injection,the quadriceps femoris injection and the intranasal irrigation respectively. All the mice were immunized two times. The immune interval was two weeks. Saliva and serum samples were collected respectively at 0, 1, 2, 4 weeks after immunization. The specific antibodies were detected by ELISA. Results:The specific antibodies were up at one week after immunization in different routes. The peak time of the antibodies level appeared at 4 weeks.The level of salivary specific anti- PAc IgA induced by submandibular gland- target injection and that of serum IgG induced by thigh bone muscle injection was the highest, respectively. The differences of antibodies level between in experiment groups and negative control group or vacant comparison group were significant(P<0.01). Conclusion: The vaccine gene pcDNA3- PAc effectively induce local mucosal immune response and systemic immune response.