Effects of Danggui-Shaoyao-San on the Influence of Spatial Learning and Memory Induced by Experimental Tooth Movement / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The pain caused by orthodontic treatment has been considered as tough problems in orthodontic practice. There is substantial literature on pain which has exactly effected on learning and memory; orthodontic tooth movement affected the emotional status has been showed positive outcomes. Danggui-Shaoyao-San (DSS) is a Traditional Chinese Medicine prescription that has been used for pain treatment and analgesic effect for orthodontic pain via inhibiting the activations of neuron and glia. We raised the hypothesis that DSS could restore the impaired abilities of spatial learning and memory via regulating neuron or glia expression in the hippocampus.</p><p><b>METHODS</b>A total of 36 rats were randomly divided into three groups: (1) Sham group (n = 12), rats underwent all the operation procedure except for the placement of orthodontic forces and received saline treatment; (2) experimental tooth movement (ETM) group (n = 12), rats received saline treatment and ETM; (3) DSS + ETM (DETM) group (n = 12), rats received DSS treatment and ETM. All DETM group animals were administered with DSS at a dose of 150 mg/kg. Morris water maze test was evaluated; immunofluorescent histochemistry was used to identify astrocytes activation, and immunofluorescent dendritic spine analysis was used to identify the dendritic spines morphological characteristics expression levels in hippocampus.</p><p><b>RESULTS</b>Maze training sessions during the 5 successive days revealed that ETM significantly deficits in progressive learning in rats, DSS that was given from day 5 prior to ETM enhanced progressive learning. The ETM group rats took longer to cross target quadrant during the probe trial and got less times to cross-platform than DETM group. The spine density in hippocampus in ETM group was significantly decreased compared to the sham group. In addition, thin and mature spine density were decreased too. However, the DSS administration could reverse the dendritic shrinkage and increase the spine density compared to the ETM group. Astrocytes activation showed the opposite trend in hippocampus dentate gyrus (DG).</p><p><b>CONCLUSIONS</b>Treatment with DSS could restore the impaired abilities on ETM-induced decrease of learning and memory behavior. The decreased spines density in the hippocampus and astrocytes activation in DG of hippocampus in the ETM group rats may be related with the decline of the ability of learning and memory. The ability to change the synaptic plasticity in hippocampus after DSS administration may be correlated with the alleviation of impairment of learn and memory after ETM treatment.</p>

Inhibition of bone marrow mesenchymal stem cell autologous transplantation on inflammation following rabbit corneal alkali burn / 中华实验眼科杂志

Background Ocular alkali burns leads to corneal ulcer and angiogenesis and even corneal opacity.There is still no ideal treatment method.Studies showed that mesenchymal stem cells (MSCs) can repair corneal wound in vivo,but the specific mechanism is still not clear.Objective This study aimed to observe the histopathological change after the early transplatation of bone marrow MSCs (BMSCs) for corneal alkali burn model in rabbits and explore the anti-inflammatory effects of MSCs after corneal alkali burn.Methods Bone marrow of 4 ml was collected from 2-3 month-old Japanese rabbit.BMSCs were isolated and cultured from the bone marrow of rabbits,and the third generation of cells were used in this study.Cultured cells were identified by morphology and the expressions of surface markers.Corneal alkali burn models were extablished in the right eyes of 24 rabbits by attaching the filter paper with 0.1％ NaOH at the central cornea for 30 seconds,and then the models were randomized into 2 groups.BMSCs suspension of 300 μl (concentration 5×l06/μl) was subconjunctivally injected 1 hour after modeling in the BMSCs group,and equal volume of PBS was used in the same way in the PBS group.Corneal opacification was scored under the slim lamp microscope in 3,14 and 28 days after injection.The polymorphonuclear neutrophils (PMNs) were counted by histopathological examination,and the expression of matrix metalloproteinase-2 (MMP-2) in the corneal tissue was evaluated by immunochemistry in various time points.The use and care of the rabbits followed the statement of ARVO.Results The rabbit BMSCs were plastic-adherent cells that exhibited a fibroblastlike shape.Cultrued cells highly expressed surface adhesion molecular markers CD29 and CD90 (99.18％ and 97.94％) and lowly expressed hematopoietic cell markers CD34 and CD31 (0.74％ and 0.15％).Opacification of cornea,defect of corneal epithelium,stromal edema and neovascularization appeared after modeling.In 14 days and 28 days after modeling,the opacification scores in the BMSCs group were 2.37±0.52 and 2.25±0.50,which were significantly lower than 3.00±0.53 and 3.25 ±0.50 in the PBS group (t =2.376,2.828,both at P＜0.05).After subconjunctival injection,the number of PMNs was (34.17 ±1.85) /12 fields and (25.64 ±3.86)/12 fields in the BMSCs group,showing significant decrease in comparison with (42.70 ±1.54) /12 fields and (32.67 ±1.42)/12 fields in the PBS group (t=10.021,4.832,both at P=0.000).The expression levels of MMP-2 (A value) in cornea were 0.388±0.016 and 0.384±0.006 in the BMSCs group,with considerable decreases in comparison with 0.438± 0.006 and 0.412± 0.005 in the PBS group (t=10.205,13.514,both at P=0.000).Conclusions Early transplantation of BMSCs can arrest the occurrance of corneal ulcer by suppressing the infiltration of PMNs,alleviateing the inflammation reaction,downregulating the expression of MMP-2 in cornea and inhibiting the degradation of stromal collagen fibers.

Role of insulin-like growth factor-1 and insulin-like growth factor-1 receptor in the migration of lens epithelial cells / 中华实验眼科杂志

Background Posterior capsular opacification (PCO) following the extracapsular extract of cataract is associated with the proliferation and migration of residual lens epithelial cells (LECs).Study showed that the incidence of PCO is higher in diabetic patients than those of non-diabetes.So if insulin-like growth factor-1 (IGF-1) participates in the pathogenesis of PCO deserve research.Objective This study was to explore the active mechanism of IGF-1/IGF-1 receptor (IGF-1 R) system in the migration of LECs and offer theoretical basis for clinical prevention and treatment of PCO.Methods Human lens epithelial cell lines (HLEC-B3) were cultured and passaged in DMEM.The cells were identified using fluorescence immunocytometry.IGF-1 with the concentrations of 0,30,90 μg/L were added into the medium separately for 48 hours.The numbers of migrated cells were calculated by Transwell test.The cells were cultured in DMEM containing 0,1.5,30,60,90 μg/L IGF-1,and the expressions of IGF-1 Rα and IGF-1Rβ in the cells were assayed and compared by Western bolt.Results The cultured showed the positive response for α-crystallin anibody with red fluorescence in the cellular membrane.Twelve hours after Transwell incubation,the number of migrated cells (Median) was 0(0,1),10(10,11) and 29(27,31) in the 0 μg/L IGF-1 group,30 μg/L IGF-1 group and 90 μg/L IGF-1 group,respectively,showing a significant difference among the 3 groups (Z=12.610,P=0.002).The number of migrated cells in the 30 μg/L IGF-1 group and 90 μg/L IGF-1 group was significantly more than that of the 0 μg/L IGF-1 group (both at P =0.008),and the number of migrated cells in the 90 μg/L IGF-1 group was significantly more than that of the 30 μg/L IGF-1group (P =0.009).Western blot assay showed that the expressions of IGF-1Rα and IGF-1Rβ in the cells were significantly different among the 0,1.5,30,60,90 μg/L IGF-1 groups (F=63.700,130.530,both P =0.000).The expressions of IGF-1 Rα and IGF-1Rβ were gradually elevated as increase of IGF-1 doses when then concentration of IGF-1 was ＞ 30 μg/L,with significant differences among the different concentrations groups (all at P＜0.05).Conclusions IGF-1 can upregulate the expressions of IGF-1R in HLEC-B3 cells in vitro in a dose-dependent manner.Also,IGF-1 enhances the migration ability of HLEC-B3 cells.These results suggest that activation of IGF-1/IGF-1R system may be associated with the pathogenesis of PCO.

Danggui-shaoyao-san, a traditional Chinese medicine prescription, alleviates the orthodontic pain and inhibits neuronal and microglia activation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The pain caused by orthodontic treatment has been considered as tough problems in orthodontic practice. Danggui-shaoyao-san (DSS) is a traditional Chinese medicine (TCM) prescription which has long been used for pain treatment and possesses antioxidative, cognitive enhancing and antidepressant effects. We raise the hypothesis that DSS exerts analgesic effect for orthodontic pain via inhibiting the activations of neuron and microglia.</p><p><b>METHODS</b>DSS was given twice a day from day 5 prior to experimental tooth movement (ETM). Directed face grooming and vacuous chewing movements (VCM) were evaluated. Immunofluorescent histochemistry and Western blot analysis were used to quantify the Iba-1 (microglia activation) and Fos (neuronal activation) expression levels in the trigeminal spinal nucleus caudalis (Vc).</p><p><b>RESULTS</b>ETM significantly increased directed face grooming and VCM which reached the peak at post-operative day (POD) 1 and gradually decreased to the baseline at POD 7. However, a drastic peak increase of Fos expression in Vc was observed at 4 hours and gradually decreased to baseline at POD 7; while the increased Iba-1 level reached the peak at POD 1 and gradually decreased to baseline at POD 7. Furthermore, pre-treatment with DSS significantly attenuated the ETM induced directed face grooming and VCM as well as the Fos and Iba-1 levels at POD 1.</p><p><b>CONCLUSION</b>Treatment with DSS had significant analgesic effects on ETM-induced pain, which was accompanied with inhibition of both neuronal and microglial activation.</p>

Effects and molecular mechanism of nicotine on odontoblasts / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the effects of nicotine on the proliferation of odontoblasts and explore the possible mechanism.</p><p><b>METHODS</b>Odontoblasts MDPC-23 were cultured, inoculated and divided into two groups randomly. With no stimuli added for the control group, the experimental group was stimulated by 100 microg/mL nicotine. After 8 hours, 10 micromol/L BrdU was added to label cells at S stage in cell cycle. 24 hours later, odontoblasts were fixed and immunofluorescence staining was performed with specific mouse BrdU antibody. After counterstaining with propidium iodide, BrdU positive cells were arbitrarily scored microscopically by an independent estimation conducted three times, and the corresponding total cell number in the same vision were counted in both groups. BrdU positive cell rates were calculated and compared statistically. At the same time, odontoblasts MDPC-23 were cultured and stimulated by 100 microg/mL nicotine, the dynamic Ca2+ concentration inside the cytoplasm were detected immediately by a confocal laser scanning microscope.</p><p><b>RESULTS</b>The ratio of S stage cells in the experimental group was 36.3% significantly lower than that (48.2%) in the control group. After the addition of 100 microg/mL nicotine, the Ca2+ concentration inside the cytoplasm rose rapidly, sustained at a high level for a short time and then relapsed gradually.</p><p><b>CONCLUSION</b>Nicotine had inhibitory effects on the proliferation of odontoblasts MDPC-23, which might be related to the increased Ca2+ concentration in the cytoplasm.</p>

Expression, subcellular localization and nuclear translocation of transcription factor up stream stimulatory factor-1 in odontoblasts / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To examine the expression and subcellular localization of transcription factor USF1 in odontoblasts and investigate whether nuclear translocation occurs under stimuli.</p><p><b>METHODS</b>Odontoblasts MDPC-23 were cultured on coverslips and divided into 2 groups. Group 1 received no stimuli, and group 2 was stimulated by nicotine with various concentrations respectively for 1h. Then the mountings of odontoblasts were prepared and immunocytochemical staining was performed with specific USF1 antibody via SABC method. Hela cells were used as positive control.</p><p><b>RESULTS</b>The staining was positive in the cytoplasm of odontoblasts in group 1, but in the nuclei of Hela cells and in 100 mg/L nicotine-stimulated odontoblasts in group 2.</p><p><b>CONCLUSIONS</b>There exists USF1 protein in odontoblasts, which locates in the cytoplasm and could translocate into nuclei under the stimulation of nicotine.</p>

Regulation of osteopontin expression in odontoblasts by upstream stimulatory factor 1 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the regulation effects of upstream stimulatory factor 1 (USF1) on osteopontin expression in odontoblasts.</p><p><b>METHODS</b>Odontoblast MDPC-23 was cultured and stably transfected with PCMV-USF1 or A-USF plasmids. Total RNA was extracted and osteopontin expression examined by semi-quantitative RT-PCR. Gray value of osteopontin was measured and statistic analysis performed.</p><p><b>RESULTS</b>Clones of stable PCMV-USF1 and A-USF plasmids transfection were obtained. Compared with the control, osteopontin was upregulated in PCMV-USF1 transfection group, and downregulated in A-USF transfection group.</p><p><b>CONCLUSIONS</b>Upstream stimulatory factor 1 could regulate the osteopontin expression in odontoblasts, which could be blocked partly by A-USF.</p>

Expression and localization of upstream stimulatory factor 1 in mice teeth development / 实用口腔医学杂志

Objective:To detect the expression of upstream stimulatory factor 1(USF1) and its tempo-spatial distribution during mice teeth development. Methods: Total protein was extracted from P1 and P11 d mice first molar teeth germ, and Western blot for USF1 was undertaken. Paraffin sections of first molar teeth germs from E13, 16, 19, P1, 5, 8, 11, 21 d and 6-month-old adult mice were prepared respectively and immunohistochemical staining was carried out. Results: Western blot analysis identified one Mr 43 000 protein from P11 d mice teeth germs, but none from P1d mice. Immunohistochemically, evidently positive staining for USF1 in mice teeth germs began from P5 d, and extended to P11 d, which was mainly confined to the cytoplasm of secreting ameloblasts and odontoblasts, but no staining in bud, cap and early bell stages of tooth germ. However, after tooth eruption on P21 d, USF1 became negative again, although it was still positive in the adjacent muscles, and the same result was observed in adult mice tooth. Conclusion: USF1 is expressed in tooth germ, which localizes solely in secreting ameloblasts and odontoblasts, and its expression was quite dynamic during mice tooth development.

The effects of three fluoride-containing agents on the remineralization of deciduous teeth in vitro / 实用口腔医学杂志

Objective: To compare the effects of three fluoride-containing agents on the remineralization of deciduous teeth in vitro. Methods: 36 deciduous teeth were randomly divided into four groups with 9 in each. The teeth were fenestrated on the labial middle surface and the rest parts of the teeth were covered with red nail polish.Every exposed area was etched,rinsed and puffed.Then the exposed areas were treated with 100 g/L(NH 4) 2MoO 2F 4(group ①) , 380 g/L Ag(NH 3) 2F (group ②) and APF-LaCl 3(group ③) for 3 minutes respectively, those in group④ were treated with deionized H 2O as the controls.Then,the teeth were placed into a bottle containing 5 ml remineralization solutions. 4 days later, the concentration of Ca 2+ in the remineralization solutions was measured. Results: The Ca 2+ concentrations (mmol/L) in the remineralization solution of group ①,②,③ and ④ were 0.338 9?0.116 5,0.570 0?0.145 5,0.517 8?0.084 8 and 0.120 4?0.046 8 respectively (P0.05:②vs③). Conclusion: The results indicate that all of the three fluoride-containing agents can promote the remineralization of deciduous teeth in vitro, and that both APF-LaCl 3 and 380 g/L Ag(NH 3) 2F are more effective than 100 g/L(NH 4) 2, MoO 2 F 4.