BACKGROUND:As a degradable scaffold material for bone tissue engineering,lactic acid is widely used in tissue regeneration and repair research,and plays an important role in promoting tissue healing,new bone formation and angiogenesis. OBJECTIVE:To observe the effect of lactic acid degradation products on osteoclasts and to investigate the effects of lactic-interfered osteoclast conditioned medium on the proliferation,migration and tube-forming capacity of human umbilical vein endothelial cells. METHODS:(1)The mouse monocyte macrophage cell line RAW264.7 at logarithmic growth period was selected,and adherent cells were cultured in the osteoclast induction medium(DMEM medium with nuclear factor-κB receptor-activating factor ligand and 10%fetal bovine serum)containing different concentrations of lactic acid(0,5,10,20 mmol/L).After 5 days of culture,tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining were conducted.After 24 hours of culture,RT-PCR was used to detect the mRNA expression of tartrate-resistant acid phosphatase 5.(2)RAW264.7 cells at logarithmic growth period were selected and adherent cells were divided into two groups.Control group was cultured in the osteoclast induction medium,while experimental group was cultured in the osteoclast induction medium containing 10 mmol/L lactic acid.After 5 days of culture,the medium in each group was removed and the cells in the two groups were cultured in the serum-free DMEM medium for another 24 hours.Cell supernatant was then collected and used as the conditioned medium after mixed with an equal volume of DMEM medium containing 10%fetal bovine serum.Human umbilical vein endothelial cells at the logarithmic growth phase were taken and separately co-cultured with the conditioned medium of the control and experimental groups.The proliferation,migration and tube-forming ability of human umbilical vein endothelial cells were observed by cell counting kit-8 assay,migration assay,scratch assay and tube-forming assay.The mRNA and protein expression of angiogenesis-related genes and proteins were observed by RT-PCR and western blot. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining showed that 5 and 10 mmol/L lactic acid promoted osteoclastic differentiation of RAW264.7 cells and the promoting effect of 10 mmol/L lactate was more significant.RT-PCR results showed that the expression of tartrate-resistant acid phosphatase-5 mRNA of osteoclast-related genes was the highest when the lactic acid concentration was 5,10,and 20 mmol/L(P<0.05),especially 10 mmol/L.Compared with the control group,the proliferation,migration and tube-forming abilities of human umbilical vein endothelial cells were significantly increased in the experimental group(P<0.05).Compared with the control group,the expression levels of vascular endothelial growth factor and angiogenin 1 mRNA and protein were increased in the experimental group(P<0.05).To conclude,lactate-induced osteoclast conditioned medium could promote the angiogenesis of endothelial cells,and the mechanism may be related to the promotion of the expression of vascular endothelial growth factor and angiogenin 1.

BACKGROUND:The increase in multi-drug resistant bacterial infections has become a major problem in modern healthcare due to the development of bacterial resistance to antibiotics and the development of new antibacterial alternative drug materials is of great importance. OBJECTIVE:To synthesize and perform a series of characterization of a CeO2 nanoenzyme to investigate its biocompatibility and antibacterial properties against Escherichia coli. METHODS:CeO2 nanoenzymes were synthesized using a hydrothermal method.The morphology,product composition,and chemical composition were analyzed using characterization methods such as X-ray diffraction,X-ray photoelectron spectroscopy,Fourier infrared analysis,Raman spectroscopy,scanning electron microscopy,and transmission electron microscopy.The peroxide-mimetic enzyme activity of CeO2 nanoenzymes was characterized using TMB color development assay.The toxic effect of CeO2 nanoenzymes at different concentrations(10,25,and 50 μg/mL)on mouse fibroblast L929 cells was evaluated using the CCK-8 assay.The antibacterial properties of CeO2 nanoenzymes against Escherichia coli under different conditions were evaluated using the plate coating method.Changes in intra-bacterial reactive oxygen species after treatment with different conditions were detected using a reactive oxygen species detection kit. RESULTS AND CONCLUSION:(1)The morphology of the synthesized CeO2 nanoparticles was rod-shaped,with Ce3+ accounting for 29.87%of the total Ce3+/Ce4+ and an average grain size of 7.4 nm.In a slightly acidic environment containing TMB and pH=5.5,CeO2 nanoenzymes mixed with H2O2 showed excellent peroxidase activity,but did not show peroxidase simulated activity at pH=7.4.(2)There was no statistically significant difference in the toxic effects of CeO2 nanoparticles at various mass concentrations on mouse fibroblast L929 cells.(3)In a slightly acidic environment at pH 5.5,Escherichia coli was inhibited to a certain extent in the presence of CeO2 nanoenzyme alone at a concentration of 10 μg/mL,with a decrease in CFU results of about 0.5 log(P＜0.01);in a slightly acidic environment containing 50 μmol/L H2O2,CeO2 nanoenzyme showed excellent antibacterial effects against Escherichia coli,with a decrease in Escherichia coli CFU results of by about 1.5 log(P＜0.001).After CeO2 nanoenzymes interacted with Escherichia coli,the level of reactive oxygen species in Escherichia coli increased(P＜0.05);after CeO2 nanoenzymes interacted with Escherichia coli together with H2O2,the level of reactive oxygen species in Escherichia coli increased significantly(P＜0.001).(4)The results show that the CeO2 nanoenzymes have good biocompatibility,are inherently antibacterial,and can exhibit peroxidase activity in a slightly acidic environment containing low concentrations of H2O2,and generate reactive oxygen species to kill bacteria,thus showing excellent antibacterial effects.

BACKGROUND:Three-dimensional(3D)printing is an emerging technology in the field of dentistry.It utilizes a layer-by-layer manufacturing technique to create scaffolds suitable for periodontal tissue engineering applications.Tissue scaffolds produced through 3D printing can possess controlled characteristics,including internal structure,porosity,and interconnectivity,making it an ideal strategy for periodontal tissue engineering. OBJECTIVE:To review the applications of 3D printed scaffolds in periodontal regeneration. METHODS:English search terms were"3D printing,periodontal tissue engineering,additive manufacturing,regenerative medicine,bioengineering,scaffold,bioprinting,periodontitis".Chinese search terms were"3D printing,additive manufacturing,periodontal tissue engineering,scaffolds,bio-inks,bioprinting,tissue engineering".Relevant literature published from 2000 to 2023 in PubMed and CNKI databases was retrieved and included in the review. RESULTS AND CONCLUSION:Over the past few decades,3D printing technology has made significant progress and breakthroughs in tissue engineering and biomedical fields.3D printing technology can provide highly personalized treatment programs,improve the suitability and therapeutic effect of therapeutic stents,and has broad application prospects in periodontal tissue engineering.In periodontal tissue engineering,3D printing applications can better mimic the complex structures of biological tissues and manufacture biocompatible scaffold materials with suitable mechanical and rheological properties.The layer-by-layer construction of tissue engineering scaffolds through 3D printing not only enables the creation of precise and intricate scaffold models for personalized treatment of periodontal disease but also facilitates the incorporation of complex microstructures and channels within the scaffolds to promote cell growth and tissue regeneration.

Objective:To investigate the expression and diagnostic values of CD200 and insulinoma associated protein 1 (INSM1) in gastrointestinal and pancreatic neuroendocrine neoplasm (GIP-NEN).Methods:The expression of CD200, INSM1, Syn and CgA was detected in 69 cases of GIP-NEN, 66 cases of gastrointestinal and pancreatic non-neuroendocrine neoplasm (GIP-nonNEN) and 16 cases of metastatic neuroendocrine neoplasm by immunohistochemistry, to compare the values of CD200, INSM1, Syn, CgA and their combinations in diagnosing GIP-NEN. Receiver operating characteristics (ROC) curve was used.Results:The immunoreactivity of CD200 was present in the cytoplasma and/or membrane of the neoplasms cells, the positive expression rates in GIP-NEN and GIP-nonNEN were significantly different ( P<0.01). The sensitivity and specificity of CD200 for diagnosing GIP-NEN were 95.7% and 78.8%, respectively. There was significant difference of the positive rates of CD200 between neuroendocrine tumor and neuroendocrine carcinoma ( P=0.05). The immunoreactivity of INSM1 was present in the nuclei of neoplasms cells. The positive expression rates in GIP-NEN and GIP-nonNEN were significantly different ( P<0.01). The sensitivity and specificity of INSM1 for diagnosis of GIP-NEN were 85.5% and 95.5%, respectively. There were also significantly different positive rates of INSM1 between neuroendocrine tumor and neuroendocrine carcinoma, as well as between G1 and G3 neuroendocrine tumors ( P<0.05). There was no difference in the area under ROC curve (AUC) of single stain of CD200, INSM1, Syn or CgA (0.857, 0.907, 0.890 and 0.833, respectively, P>0.05). The sensitivity of combined CD200+INSM1 stains for diagnosing GIP-NEN was significantly higher than that of Syn+CgA (85.5% vs. 63.8%, P<0.05). The AUC of two combinations were 0.962 and 0.925, respectively, which were not statistically different ( P>0.05). Conclusions:CD200 and INSM1 are two novel markers of neuroendocrine neoplasm, which aid to diagnosis for GIP-NEN and exclude its mimickers. They are associated with tumor grades. Combining both as an immunohistochemical panel shows high sensitivity and specificity. Thus, the combined panel can be utilized as useful supplement for Syn and CgA.

BACKGROUND: Hedgehog signaling pathway is shown to contribute to the osteogenesis of bone marrow mesenchymal stem cells, and to play an important role in regulating the expression of RANKL in osteoblasts. Nowadays, the definite signal transduction pathway has been revealed gradually.OBJECTIVE: To review the research progress of RANKL in osteoblasts that regulated by hedgehog signaling pathway.METHODS: A computer-based online search in CNKI, Google Scholar and PubMed databases was performed with the key words of Hedgehog, osteoblast, BMSCs, PTHrP, CREB, NFAT, RANKL in English and Chinese, respectively.Literatures related to the expression of RANKL in osteoblasts that regulated by the Hedgehog signaling pathway were included initially and 37 eligible articles were extensively summarized for review.RESULTS AND CONCLUSION: The Hedgehog signaling pathway plays an advanced role in the osteogenic differentiation of bone marrow mesenchymal stem cells, and also upregulates intracellular parathyroid hormone related protein, which activates its downstream signaling molecules cAMP response element binding protein and nuclear factor of activated T cells ulteriorly, to promote the expression of RANKL in osteoblasts and increase the differentiation and formation from osteoclast precursor cells to mature osteoclasts.

BACKGROUND: Recently, zirconia ceramic implants have been reported to be used in clinic. Increasing studies on surface treatment of ziconia implants have been done, in order to promote bone-implant osseointegration and increase the success rate of implantation. There are no recognized surface treatment techniques of zirconia implants as yet.OBJECTIVE: To summarize the research progress in surface treatment techniques of zirconia implants.METHODS: The first author searched the CNKI, PubMed and Web of Science databases for relevant articles published from January 2000 to October 2016. The keywords were zirconia or zirconium, implant, surface treatment in Chinese and English, respectively. The search condition was((zirconia or zirconium) AND implant) AND surface treatment. Here,we reviewed the surface treatment techniques of zirconia implants, and introduced the properties, the surface treatment techniques and commercialization of zirconia implants.RESULTS AND CONCLUSION: Totally 23 papers were included in result analysis. Commonly used surface treatment technologies of zirconia implants include sand blasting, acid etching, sandblast and acid-etching, selective filtration etching, laser, ultraviolet and coating. These technologies all contribute to improving the surface activity of the implant,increasing the roughness, and promoting bone bonding to a certain extent.Because of the very limited experiments,further investigations are required to explore the optimum technique for the surface treatment of zirconia implants.

Objective:To research the best processing method of ginger juice baking for Rhizoma coptis. Methods:The total con-tent of four alkaloids including berberine hydrochloride determined by HPLC was used as the evaluation index, and an L9(34)orthogo-nal design with three factors including the amount of ginger juice, baking temperature and baking time and variance analysis were ap-plied to study the ginger juice baking technology for Rhizoma coptis. Results:The best processing conditions were as follows:Rhizoma coptis was soaked with 15% ginger juice, baked at 150℃ for 40 min, and withdrawn to be cool. Conclusion:The optimal ginger juice baking technology for Rhizoma coptis is reasonable, which can be used to guide the standardized production of Rhizoma coptis with gin-ger juice baking.

OBJECTIVE:To investigate the effect of Changji’an capsules on abdominal pain and the molecular mechanism re-lated to calcitoningene-related peptide(CGRP)and corticosterone(CORT)in diarrhea-predominant irratable bowel syndrome(IBS-D)model rats. METHODS:After the rat models of IBS-D were established by the method of separation of breast mick combined with stimulation with acetic acid,the rats were randomly divided into model group(isometric normal saline),pinaverium bromide group(0.018 g/kg),and Changji’an capsules high,medium and low dose groups(2.812 g/kg,1.406 g/kg and 0.703 g/kg),and another SD rats were included in the normal control group(isometric normal saline). The drugs were given to the rats once a day for consecutive 14 d,ig. Injection of normal saline method was adopted to determine the rat’s sensibility to abdominal pain. The en-zyme-linked immunosorbent assay(ELISA)was adopted to determine the content of CORT in the serum of the rats. Subjected to re-verse transcription polymerse chain reaction(RT-PCR)was adopted to determine the expression of CGRP mRNA in the hypothala-mus and colon tissues of the rats. RESULTS:Compared with normal control group,threshold values of arching the back and stick-ing out the abdomen were decreased,the content of CORT in serum and expression of CGRP mRNA in the hypothalamus and co-lon tissues in model group were increased,with significant difference(P<0.01). Compared with model group,threshold values of arching the back and sticking out the abdomen were increased,the content of CORT in serum and expression of CGRP mRNA in the hypothalamus and colon tissues in pinaverium bromide group and Changji’an capsules high and medium dose groups were de-creased;the threshold value of arching the back in Changji’an capsules low dose group were increased,with significant difference (P<0.01 or P<0.05). CONCLUSIONS:Changji’an capsules can improve the abdominal pain in rats with IBS-D by a mechanism that may be related to the decrease in the expression of CGRP mRNA in the hypothalamus and colon tissues and the reduc-tion of the content of CORT in serum.

Objective To observe the effects of low-dose theophylline and tiotropium on lung function and quality of life in patients with mild-moderate stable chronic obstructive pulmonary disease (COPD) and evaluate its safety.Methods This was a randomized,parallel-group,controlled trial.A total of 115 patients with mild-moderate stable COPD were divided into tiotropium group (37 cases),slow-release theophylline group (40 cases) and the combination of slow-release theophylline and tiotropium group (38 cases) by random digits table method.Thirty-eight patients without cardiopulmonary diseases were enrolled in control group.Observation period was 12 months.The lung function,6 min walking test distance (6MWD),modified British Medical Research Council Scale (mMRC),and COPD assessment test (CAT) were monitored before treatment and after treatment for 12 months.Results Of 115 patients,107 patients (35cases in tiotropium group,36 cases in slow-release theophylline group and 36 cases in combination of slowrelease theophylline and tiotropium group) completed the study.No significant difference was found in spirometry parameters reflecting airflow limitation after 12 months treatment compared with that before treatment in tiotropium group,slow-release theophylline group and the combination of slow-release theophylline and tiotropium group (P ＞ 0.05),such as the percentage of forced expiratory volume in 1 second (FEV1) over the expected value (FEV1％) and FEV1/forced vital capacity (FVC).The percentage of mid expiratory flow over the expected value (FEF25～75％) was improved in all groups,but the increment of FEF25～75 ％ was much higher in tiotropium group and the combination of slow-release theophylline and tiotropium group than that in slow-release theophylline group:(39.23 ± 7.77)％,(39.99 ± 8.25)％ vs.(34.91 ± 9.50)％,there were significant differences (P ＜ 0.05).Similar changes were observed in mMRC and CAT score.There was significant difference in CAT score between tiotropium group,the combination of slow-release theophylline and tiotropium group and slow-release theophylline group:(14.34 ± 2.22),(14.39 ± 3.53) scores vs.(16.22 ± 3.35) scores,P ＜ 0.05.6MWD was no obvious change in tiotropium group,slow-release theophylline group and the combination of slow-release theophylline and tiotropium group before and after treatment.The use frequency of short-acting drugs bronchiectasis was the lowest in the combination of slow-release theophylline and tiotropium group,and only was (2.3 ± 1.4) times per week.Fourteen patients happened COPD exacerbations in slow-release theophylline group during 12 months treatment.The duration in slow-release theophylline group was more than that in tiotropium group and the combination of slow-release theophylline and tiotropium group:(9.76 ± 2.25) d vs.(7.85 ± 2.51),(8.29 ± 2.24) d,and there was significant difference (P ＜ 0.05).Conclusions For mild-moderate stable COPD patients,the quality of life and dyspnea scores are not improved significantly after treatment of low-dose slow-release theophylline,and the combination slow-release theophylline and tiotropium may be more beneficial and safe.

Objective To investigate the feasibility of extraction site preservation using injectable calcium phosphate cement (CPC) combine with poly (lactic-co-glycolic acid) (PLGA) microspheres.Methods Immediate extraction defects models were created in canine mandibles,and the defects were filled with CPC/PLGA (experimental group,E),Bio-Oss (positive control,P),non-treatment (blank control,B) respectively.Dogs were sacrificed after 4,8,12 weeks post operation.Statistical analysis were conducted using SPSS 19.Results Results of radiological observation showed that there were not significantly different between groups in 4 and 8 week(P ＞0.05).After 12 week,E(114.9 ±8.4)were not significantly different compared with P(117.4 ± 12.1) (P ＞ 0.05),both were significantly higher than B (95.0 ± 12.6)(P ＜0.05).Histology examination showed that at 4 week following surgery,the result of newly formed bone was as follow,P [(87.5 ± 1.5) ％] ＞ B [(78.7 ± 2.7) ％] ＞ E [(69.2 ± 1.8) ％] (P ＜ 0.05).At8,12 week,results of P[(94.0 ±2.3)％ and (93.5 ±1.9)％]and E[(94.7 ±1.1)％ and (96.0±0.9)％] were better than those of B[(76.8 ±3.0)％ and (87.0 ±2.4)％] (P＜0.05).Conclusions The effect of CPC/PLGA repair immediate alveolar ridge defects is the same as that of BioOss,and CPC/PLGA can be used as a material in extraction site preservation.