凋亡相关蛋白在激素性幼兔股骨头缺血坏死动物模型中的表达及意义

目的 观察各凋亡相关蛋白在激素性幼兔股骨头缺血坏死动物模型股骨头组织中的表达情况,探讨该模型中调控凋亡的主要通路.方法 选用2月龄的新西兰大白兔,制作糖皮质激素性幼兔股骨头缺血坏死模型及对照组模型,根据是否发病将激素注射组分为未发病组和发病组.取股骨头软骨及软骨下骨组织用免疫组织化学法检测凋亡通路中凋亡相关蛋白天冬氨酸特异酶切的半胱氨酸蛋白酶-3(Caspase-3)、Caspase-8、人结合凋亡蛋白酶活化因子-1(apaf-1)、钙蛋白酶-1(calpain-1)的表达情况.分别测定在单位视野中Caspase-3、Caspase-8、apaf-1、calpain-1的积分光密度(IOD)值.结果 1.Caspase-3的IOD值分别为发病组25 142.72 ±21 528.48,未发病组2 069.63±1 096.96,对照组301.80±99.66.Caspase-8的IOD值分别为发病组24 942.63±18 942.99,未发病组2016.31±1 518.70,对照组236.85±97.94.Apaf-1的IOD值分别为发病组8 514.23±6 384.20,未发病组1 118.49±1 360.59,对照组95.13±38.05.Calpain-1的IOD值分别为发病组9 636.71 ±9 123.81,未发病组1 881.10±3 277.86,对照组126.71±47.35.Caspase-3在发病组和未发病组、对照组间差异有统计学意义(H=11.470、23.996,p＜0.01);Caspase-8在发病组和对照组间差异有统计学意义(H=22.178,P＜0.01);apaf-1在发病组和对照组间差异有统计学意义(H=22.808,P＜0.01);calpain-1在发病组和对照组间差异有统计学意义(H=13.553,P ＜0.01).2.线性回归分析:Caspase-8能够显著的预测Caspase-3,且回归方程回归效应显著,回归方程能够解释40.3％的变异,而apaf-1和calpain-1对Caspase-3的回归效应不显著.结论 凋亡受体通路可能在股骨头缺血坏死的凋亡过程中发挥主要调控作用.     

Side‐Effects of Irinotecan (CPT‐11), the Clinically Used Drug for Colon Cancer Therapy,Are Eliminated in Experimental Animals Treated with Latex Proteins from Calotropis procera (Apocynaceae)

Intestinal mucositis (IM) is the critical side effect of irinotecan (CPT‐11), which is the front‐line drug used for the treatment of colorectal cancer. This study aimed to evaluate the effectiveness of latex proteins (LP) from Calotropis procera to prevent IM and diarrhea in animals. Swiss mice were treated daily with saline or LP (1, 5, or 50 mg/kg, i.v.) 24 h prior to CTP‐11 (75 mg/kg/4 days, i.p) and for additional 6 days. Animal survival, body weight variation, and diarrhea were registered. After animal sacrifice (day 7 post first injection of CPT‐11), intestinal samples were collected to study morphology and inflammatory parameters. Animals given LP exhibited improved parameters (survival, body weight, and absence of diarrhea) as compared with the CPT‐11 control. The severity of IM observed in animals given CPT‐11 was reduced in animals treated with LP. Treatment with LP also prevented the reduction in the villus/crypt ratio promoted by CPT‐11. The rise in MPO activity and pro‐inflammatory cytokines, over‐contractility of the smooth muscle, and diarrhea were all abrogated in LP‐treated mice. Markedly reduced immunostaining intensity for COX‐2, TNF‐α, IL‐1β, iNOS, and NF‐κB was observed in the intestinal tissue of animals treated with LP. The side‐effects of CPT‐11 were eliminated by LP treatment in experimental animals and improved clinical parameters characteristic of IM All known biochemical pathogenesis. Copyright © 2016 John Wiley & Sons, Ltd.     

釉基质蛋白对人牙龈上皮细胞增殖的影响

目的:研究釉基质蛋白(enamelmatrixproteins,EMPs)对体外培养的人牙龈上皮细胞增殖活性的影响。方法:乙酸提取法获取EMPs,取龈切术中切除的牙龈组织,采用酶消化法培养牙龈上皮细胞。将第1代牙龈上皮细胞按照22500个/孔接种,分为4组:对照组(不加EMPs)及EMPs分别为50、100、200μg/ml的实验组。采用MTT法检测各组细胞数量,对每个时间点的各组实验数据进行方差分析。结果:人牙龈上皮细胞能在EMPs覆盖的平皿上生长。统计学分析表明,不同浓度的EMPs在早期对牙龈上皮细胞的增殖无显著影响;从第3天开始,当培养液中EMPs浓度为200μg/ml时,牙龈上皮细胞增殖显著受抑。结论:EMPs影响牙龈上皮细胞的增殖,并存在剂量和时间依赖性。     

Sinus floor augmentation with simultaneous placement of dental implants in the presence of platelet-rich plasma or recombinant human bone morphogenetic protein-7

The aim of the present study was to evaluate the possible benefit of platelet-rich plasma (PRP) in sinus grafting as compared with recombinant human bone morphogenetic protein-7 (rhBMP-7). For this purpose, we performed a bilateral sinus augmentation with anorganic bovine bone and simultaneous insertion of a titanium screw implant in five miniature pigs. Six hundred microliters of PRP and 15%-vol. autologous bone, which was collected with a trap during preparation of the implant recipient site, were added to the right sinus and 420 microl rhBMP-7 to the left sinus. A polychrome sequential labeling was performed. The animals were sacrificed 6 weeks after surgery. Undecalcified ground sections were evaluated by microradiography, digitized histomorphometry and under fluorescent light. The mean bone-implant contact using rhBMP-7 was 45.8% and 5.7% under PRP (P=0.002). The mean height of newly mineralized bone in the augmented area using rhBMP-7 amounted to 8.3 mm as opposed to 3.6 mm under PRP (P=0.013). Using PRP, the mean area of the newly formed bone was enhanced (51.3%) as compared with rhBMP-7 (33.1%); however, this difference was not statistically significant (P=0.081). In conclusion, under the selected experimental conditions the use of rhBMP-7 led to superior outcomes with regard to the osseointegration of dental implants and the height of new bone as compared with the use of PRP.     

切胶免疫制备腮腺液高丰度蛋白多克隆抗体

目的 通过切胶免疫制备人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备提供抗原解决方案.方法 将腮腺液经超滤法浓缩蛋白后,进行SDS-PAGE分析,切取分子量约为50-65kDa的高丰度蛋白条带,研磨后注射新西兰大耳白兔诱导产生免疫应答并制备兔抗人腮腺液高丰度蛋白多克隆抗体,并应用蛋白免疫印记(Western blot)等方法进行抗体鉴定.结果 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体.结论 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备及唾液蛋白质组学研究奠定基础.     

Periodontal tissue engineering and regeneration: Consensus Report of the Sixth European Workshop on Periodontology

Aim: To review the scientific preclinical background and clinical studies of current methods of periodontal regeneration in the treatment of infrabony defects and soft tissue deficiencies
Method: Five commissioned review papers including two systematic reviews were scrutinized by a group of experts in order to derive consensus conclusions, clinical relevance/implications and to propose future research requirements.
Results: The following five papers were assessed:
  1. Biological mediators and periodontal regeneration: a review of enamel matrix proteins at the cellular and molecular levels.
  2. Regeneration of periodontal tissues: combination of barrier membranes and grafting materials – Biological foundation and preclinical evidence.
  3. Clinical outcomes with bioactive agents alone or in combination with grafting or GTR
  4. Treatment of gingival recession with coronally advanced flap procedures. A systematic review.
  5. Soft tissue management at implant sites     

釉基质蛋白体外诱导大鼠外胚间充质细胞骨桥素和骨钙素mRNA的表达

目的：了解体外培养环境中，大鼠外胚间充质细胞在两种不同形式釉基质蛋白(粗提型EMPs及纯化型EMD)诱导条件下，成骨分化标志骨桥素及骨钙素mRNA的表达水平。方法：酶消化法原代培养SD大鼠颌突外胚间充质细胞，在含釉基质蛋白(EMPs或EMD或EMPs EMD)的培养液中进行体外诱导培养，提取总RNA，采用RT-PCR法，以β-actin cDNA为内参照，检测骨桥素及骨钙素mRNA的表达水平。结果：1)各诱导实验组细胞均出现骨桥素mRNA的表达；2)仅EMPs EMD组出现骨钙素mRNA的表达。结论：外胚间充质细胞经釉基质蛋白特别是粗提型EMPs及纯化型EMD体外复合和诱导培养，可向成骨或成牙骨质方向分化，出现骨桥素及骨钙素mRNA的表达。     

Separation of the outer membrane and identification of major outer membrane proteins from Porphyromonas gingivalis

The outer membrane of Porphyromonas gingivalis, an oral strict anaerobe, was isolated by sucrose density gradient centrifugation. The outer membrane obtained by the differential detergent extraction method, previously reported, showed an essentially similar protein pattern on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), confirming that the latter method is suitable for the study of outer membrane proteins in this organism. N-terminal amino acid sequence analysis revealed that major outer membrane proteins in this organism included Arg-gingipain, Lys-gingipain, RagA (a TonB-linked receptor), and putative porins that were homologous to Escherichia coli OmpA.     

Antibacterial effect of ozone on cariogenic bacterial species

Objective

The aim was to evaluate the antibacterial effect of ozone on cariogenic bacterial species with and without the presence of saliva and a possible effect on the salivary proteins.

Methods

Suspensions of Actinomyces naeslundii (ACTCC 12104^T), Lactobacilli casei (N CTC 151) and Streptococcus mutans (NCTC 10449), in salt buffer or in saliva, were exposed to ozone gas delivered by the ozone generator Healozone™ 2130C. Aliquots of the suspensions were taken after 10, 30 and 60 s ozone exposures and cultivated on agar plates. Initial number of bacteria per ml was 8.0 × 10⁷ (SD 2.2 × 10⁷) (A. naeslundii), 1.0 × 10⁸ (SD 3.1 × 10⁶) (L. casei) and 1.0 × 10⁸ (SD 7.0 × 10⁵) (S. mutans), respectively. The proteins were separated by SDS electrophoresis and visualized by silver staining.

Results

In salt buffer 92%, 73% and 64% of the initial numbers of A. naeslundii, S. mutans and L. casei, respectively, were killed already after 10 s ozone exposure, while approximately 99.9% of the bacteria were dead after a 60 s exposure. After 10 and 30 s, but not after 60 s exposure to ozone, S. mutans and L. casei were less efficiently killed in saliva compared to the salt buffer. Various saliva proteins were degraded by ozone after a 60 s exposure.

Conclusions

The cariogenic species S. mutans, L. casei and A. naeslundii were almost eliminated following 60 s of ozone treatment. This killing was reduced in the presence of saliva although increasing the ozone application time to 60 s overcame these reductants in saliva. Detection of altered salivary proteins indicates that saliva components constitute additional targets for ozone.     

A clinical study evaluating the treatment of supra-alveolar-type defects with access flap surgery with and without an enamel matrix protein derivative: a pilot study

Aim: There is evidence that regenerative treatment of intra-bony and mandibular class II furcation defects with access flap and an application of an enamel matrix protein derivative (EMD) can result in a clinical benefit compared with access flap alone. The aim of this pilot study was to check if the results of access flap surgery in suprabony defects are improved by additional application of EMD.
Material and Methods: Thirty-nine adult subjects with supra-alveolar-type defects were randomly assigned to a test ( n =25) and a control group ( n =14). Seventy teeth were treated with EMD; 28 teeth were treated by access flap. Probing depth (PD), clinical attachment level and bleeding on probing were evaluated at baseline and after 12 months.
Results: PD of the operated teeth was improved in both groups ( p <0.001 to p =0.041) but always better in the test group. The attachment gain was 2.72±1.80 mm at sites with an initial PD 7 mm in the test group and 0.78±0.62 mm in the control group ( p =0.004). In the test group the mean attachment gain was 0.97±0.92 mm ( p <0.001); the mean reduction of PD was 1.55±0.90 mm ( p <0.001).
Conclusions: The data suggest a significant clinical benefit of supplementary application of EMD during surgical treatment of periodontitis of supra-alveolar pockets, especially in deeper pockets.