Objective To investigate the effects of excessive aluminum on osteoclastic formation and function in C57BL/6 mice.Methods Six-week-old male C57BL/6 mice (n =20) were randomized by weight and divided into control group and treatment group.The mice were assigned to distilled water and 270 mg/L aluminum ion,respectively.After the 15-week treatment period,the mice were euthanized by ether asphyxiation.Concentration of serum aluminum ion was determined by inductively coupled plasma (ICP).TRAP+ cells in vivo were observed using histomorphometry and osteoclasts microstructure using electron microscope (TEM).Osteoclasts were induced by macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor κB ligand (RANKL) with bone marrow macrophages separated from femur and tibia.The expression of mRNA related with bone resorption was detected,including c-FBJ osteosarcoma oncogene (c-Fos),nuclear factor of activated T-cells cl (NFATc1),c-nonreceptor tyrosine kinase (c-Src),dentrocyte expressed seven transmembrane protein (DC-STAMP),d2 isoform of vacuolar (H1) ATPase v0 domain (ATP6v0d2) and matrix metalloproteinase 9 (MMP-9).Results The data showed that the treatment group [(12.04 ± 0.21)mg/L] had significantly increased serum aluminum ion compared with the control group [(11.00 ± 0.04)mg/L,F =10.286,P ＜ 0.05].TRAP+ cells examination confirmed osteoclasts in treatment group [(31.39 ± 9.80) number] were significantly higher than those in the control group [(5.46 ± 4.15) number,F =9.344,P ＜ 0.05].A large proportion of osteoclasts in treatment group lacked ruffled borders and showed vacuolation degeneration.The expression of mRNA in treatment group was lower than that of the control group.The expression of mRNA in control group and treatment group was NFATcl (3.25 ± 0.93 vs.0.29 ± 0.18,F =11.602,P ＜ 0.05),c-Fos (0.86 ± 0.16 vs.0.16 ± 0.02,F =9.405,P ＜ 0.05),c-Src (8.82 ± 1.51 vs.2.29 ± 0.36,F =9.128,P ＜ 0.05),DC-STAMP (3.70 ± 0.70 vs.1.36 ± 0.57,F =10.298,P ＜ 0.05),ATP6v0d2 (15.60 ± 4.81 vs.1.39 ± 0.95,F =8.828,P ＜ 0.05),and MMP-9 (18.64 ± 7.62 vs.2.10 ± 0.92,F =9.356,P ＜ 0.05),respectively.Conclusions Aluminum can increase the number of osteoclasts under epiphyseal plate,but inhibits osteoclasts differentiation.This phenomena may be related with decreased expression of c-Src,DC-STAMP,c-Fos,NFATcl,ATP6v0d2 and MMP-9 mRNA which regulate the function of osteoclasts.

In the present study,we examined the codon usage bias between pseudorabies virus (PRV) US1 gene and the US1-like genes of 20 reference alphaherpesviruses.Comparative analysis showed noticeable disparities of the synonymous codon usage bias in the 21 alphaherpesviruses,indicated by codon adaptation index,effective number of codons (ENc) and GC3s value.The codon usage pattern of PRV US1 gene was phylogenetically conserved and similar to that of the US1-like genes of the genus Varicellovirus of alphaherpesvirus,with a strong bias towards the codons with C and G at the third codon position.Cluster analysis of codon usage pattern of PRV US1 gene with its reference alphaherpesviruses demonstrated that the codon usage bias of US1-like genes of 21 alphaherpesviruses had a very close relation with their gene functions.ENc-plot revealed that the genetic heterogeneity in PRV US1 gene and the 20 reference alphaherpesviruses was constrained by G+C content,as well as the gene length.In addition,comparison of codon preferences in the US1 gene of PRV with those of E.coli,yeast and human revealed that there were 50 codons showing distinct usage differences between PRV and yeast,49 between PRV and human,but 48 between PRV and E.coli.Although there were slightly fewer differences in codon usages between E.coli and PRV,the difference is unlikely to be statistically significant,and experimental studies are necessary to establish the most suitable expression system for PRV US1.In conclusion,these results may improve our understanding of the evolution,pathogenesis and functional studies of PRV,as well as contributing to the area of herpesvirus research or even studies with other viruses.

BACKGROUND:Poultry mesenchymal stem cels are a particular subset of pluripotent adult stem cels derived from the mesoderm, which have great application prospects because of their strong proliferation and multi-directional differentiation potential. OBJECTIVE:To review the source, separation, purification, culture and differentiation of poultry mesenchymal stem cels, and to provide the theoretical foundation and experimental basis for the further research and application of poultry mesenchymal stem cels. METHODS:PubMed and CNKI databases were searched by the first author using key words of “mesenchymal stem cels, poultry, chicken, isolation, culture, differentiation” in English and in Chinese, respectively, to retrieve relevant articles published from 1990 to 2014. Literatures addressing induced poultry mesenchymal stem cels were included, and 42 articles were chosen for further analysis eventualy. RESULTS AND CONCLUSION: Poultry mesenchymal stem cels have great application prospects in the aspects of establishingin vitro model of poultry cels, studying poultry disease pathogenesis, animal nutrition and meat quality control. Its origin source is wide and easy to obtain. Isolation, purification, culture and biological characteristics of mesenchymal stem cels from different tissues are different. But, the study on poultry mesenchymal stem cels is stil in the exploration process, and there are many technical problems to be solved.

Objective To explore the distribution of serum Mg,A1,Ca,Fe,Zn and Cu ions in Tibetan population in drinking tea type fluorosis areas,and to provide a clue for pathogenesis of drinking tea type fluorosis.Methods Tibetans from six villages in Qinghai Province (Maqin County and Dari County),who were over 16 years old,born and grew up in those villages,were included.All of the participants were received epidemic questionnaire survey,tea water samples were collected and the fluoride concentration was tested based on the standard of Brick Tea Fluoride Content (GB 19965-2005).Meanwhile,the daily amount of brick tea consumption was surveyed to calculate the daily intake of tea fluoride.Blood samples were collected and the concentration of serum Mg,Al,Ca,Fe,Zn and Cu ions was tested by the method of inductively coupled plasma mass spectrometry (ICP-MS).All of the participants were diagnosed by X-ray,the parts we scheduled were forearm,shank and pelvic,then the skeletal fluorosis was diagnosed based on the Diagnostic Criteria for Endemic Fluorosis (WS/T 192-2008).Results A total of 170 people were surveyed,74 people were skeletal fluorosis,and 96 people were non-skeletal fluorosis.The median (quartiles) of daily intake of tea fluoride was 7.35 (3.00,12.30) mg.The concentration of serum Mg ion was 22.02 (17.30,23.67) mg/L,Al ion was 0.22 (0.14,0.38) mg/L,Ca ion was 100.03 (88.56,112.73) mg/L,Fe ion was 1.66 (1.26,2.36) mg/L,Zn ion was 0.80 (0.63,0.95) mg/L and Cu ion was 1.28 (0.99,1.48) mg/L.The concentration of serum Fe ion was higher in male [2.13 (1.37,3.09) mg/L] than that of female [1.56 (1.18,2.02) mg/L,Z =3.28,P ＜ 0.01].The concentration of serum Mg,Ca,Cu and Zn ions in ≥70 years old group was 15.09 (13.64,24.13),68.67 (58.67,97.24),0.97 (0.72,1.34),and 0.54 (0.48,0.74) mg/L,respectively,which was lower than those in ＜ 40 years old group [21.67 (20.08,22.76),98.71 (90.77,113.97),1.35 (1.21,1.71),and 0.78 (0.73,1.01) mg/L],the differences were statistically significant (Z =2.26,2.99,3.01,3.34,P ＜ 0.05).Fe ion in 40-49 years old group was 1.77 (1.45,3.02) mg/L,in 50-59 years old group was 1.92 (1.44,2.66) mg/L,both of them were higher than those of ＜ 40 years old group [1.34 (0.94,1.57) mg/L,Z =-3.25,-2.89,P ＜ 0.05].People whose daily intake of tea fluoride over 4 mg had a lower concentration of Ca ion [98.22 (75.48,111.22) mg/L] than people whose daily intake of tea fluoride ＜ 2 mg [110.24 (97.50,113.97) mg/L,Z =2.41,P ＜ 0.05].No significant difference was found between different degrees of skeletal fluorosis and non-skeletal fluorosis (P ＞ 0.05).Conclusions In the Tibetans who lived in the drinking tea type fluorosis areas,the concentration of serum Mg,A1,Ca,Fe,Zn and Cu ions is similar between skeletal fluorosis and non-skeletal fluorosis,which is also similar between higher daily intake of tea fluoride and lower daily intake of tea fluoride.However,it is different between older people and younger people.

Objective To investigate the relationship between single nucleotide polymorphism(SNP)of the peroxisome proliferator activated receptor γ (PPARγ) gene Rs1801282 and brick-tea type fluorosis. Methods From 2012 to 2013, this cross-sectional study was performed in 16 endemic fluorosis areas of brick-tea type in Inner Mongolia Autonomous Region,Qinghai and Xinjiang Uygur Autonomous Region of China,to select adults>18 years old as subjects, who were diagnosed as skeletal fluorosis by X-ray. All of the subjects filled in demography survey questionnaire; the survey contents included general characteristic s, and average daily brick tea intake. Drinking tea samples and urine samples of each subject were collected, and fluoride content of urine and brick-tea was determined via the ion selective electrode method (WS/T 89-2006). X-ray scintigraphy was used to diagnose skeletal fluorosis, according to the "Diagnostic Criteria of Endemic Skeletal Fluorosis" (WS/T 192-2007); the subjects were divided into skeletal fluorosis group (case group) and non-skeletal fluorosis group (control group). To collect venous blood 5 ml, whole blood DNA was extracted, and polymorphism at Rs1801282 of PPARγ was detected by MassARRAY time-of-flight mass spectrometry, to calculate odds ratio (OR) and 95% confidence interval (CI). Results There were 1 414 people included in this study,including 347 in case group and 1 067 in control group. By the Hardy-Weinberg balance test, the PPARγ gene Rs1801282 genotype was representative in case group, control group and each nationality (P > 0.05). The difference of PPARγ gene Rs1801282 genotype in case group and control group was not statistically significant (OR was 0.991, 95%CI: 0.704 - 1.395, the adjusted OR was 1.026, 95%CI: 0.707-1.489).The difference of PPARγ gene Rs1801282 genotype(CC,CG+GG)in case group and control group in different nationality was not statistically significant (Tibetan: OR was 1.400, 95%CI: 0.576 - 3.404, the adjusted OR was 1.258, 95%CI: 0.474 - 3.340; Kazak: OR was 0.898, 95%CI:0.516 -1.562,the adjusted OR was 0.936,95%CI:0.532 -1.648;Mongolia: OR was 1.148,95%CI:0.508-2.594, the adjusted OR was 1.644, 95%CI: 0.683 - 3.956; Han: OR was 1.058, 95%CI: 0.451 - 2.482, the adjusted OR was 0.959, 95%CI: 0.388 - 2.371; Russian: OR was 0.000, 95%CI: 0.000 - 0.000, the adjusted OR was 0.000, 95% CI: 0.000 - 0.000) with binary Logistic regression analysis. Conclusion We have found no association between SNP of PPARγ gene Rs1801282 and skeletal fluorosis of brick-tea type fluorosis in China.

Objective:To investigate the association of single nucleotide polymorphism at the estrogen receptor 1(ESR1) gene rs1801132 with the risk of brick-tea type skeletal fluorosis.Methods:The typical brick-tea type fluorosis areas in Qinghai, Xinjiang, and Inner Mongolia were selected as the survey sites for a cross-sectional study. An epidemiological questionnaire was conducted by the staffs on the sites for participants older than 16 years, and physical examination and X-ray diagnosis were performed. Brick tea, blood, and urine samples were collected at the same time. The diagnosis of skeletal fluorosis through X-ray was based on the "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS/T 192-2008); The determination of tea's fluoride and urinary fluoride was performed by fluoride ion-selective electrode method; gene sequencing analysis of rs1801132 locus of ESR1 gene was done by Sequenom MassARRAY flight mass spectrometry system.Results:A total of 994 patients were included in this study. The total prevalence of skeletal fluorosis was 23.9% (238/994). The prevalence of skeletal fluorosis in Tibetans(39.9%, 123/308) was higher than those of Mongolian and Han nationality [22.2% (58/261), 13.4% (57/425), χ 2=20.435, 67.811, P < 0.05]. Based on binary logistic analysis, the daily tea fluoride intake ≤ 3.5 mg, urinary fluoride content ≤1.6 mg/L, and age ≤45 years were used as the reference groups, and then, when the daily tea fluoride intake > 7.0 mg ( OR=2.865, 95% CI: 1.923-4.268), urinary fluoride content > 1.6-3.2 mg/L ( OR=2.368, 95% CI: 1.686-3.326) and > 3.2 mg/L ( OR=3.559, 95% CI: 2.401-5.276), the age > 45-65 years old ( OR=2.361, 95% CI: 1.603-3.477) and > 65 years old ( OR=4.556, 95% CI: 2.845-7.296), the risk of fluorosis was higher than that of the reference group, respectively. When the daily tea fluoride intake was > 3.5-7.0 mg and the level of urinary fluoride was > 1.6-3.2 mg/L, G allele had a protective effect on skeletal fluorosis in Mongolian population (adjusted OR=0.207, 95% CI: 0.044-0.974); when the daily tea fluoride intake was > 3.5-7.0 mg, gender was male group, G allele had a protective effect on skeletal fluorosis in Han population (adjusted OR=0.315, 95% CI: 0.112-0.887). Conclusion:The single nucleotide polymorphism of the rs1801132 locus at the ESR1 gene may be associated with the risk of susceptibility to brick-tea type skeletal fluorosis in Mongolian and Han nationality.

Fluorosis is widely prevalent worldwide, and China is one of the countries with a high incidence of endemic fluorosis. In recent years, study on non skeletal damage caused by fluorosis, especially cardiovascular system damage, has gradually increased. Fluoride can cause cardio vascular arteriosclerosis, hypertension and other diseases, while cardiovascular disease have hidden and acute onset, and the mortality rate has increased year by year in recent years. At present, the mechanism of cardiovascular diseases caused by fluoride is not yet clear, and further clarification is needed. This article provides an overview of the effects of fluoride on cardiovascular diseases from three aspect: epidemiological investigation, animals experiment, and in vitro cell experiment. It categorizes and analyzes the pathogenesis, providing new ideas for the study of cardiovascular system damage caused by fluorosis.

Objective To investigate the effect of fluoride on osteoclast in bone tissue of rats and its mechanism.Methods Twenty specific pathogen free male Wistar rats aged 3 weeks were randomly divided into two groups by weight (each group has 10).The rats of control group drink distilled water and treatment group drink distilled water containing 100 mg/L fluoride.The rats were fed for 3 month.The dental fluorosis in rats was observed.The ion selective electrode method was used to measure bone fluoride accumulation.The pathological changes of bone tissue in rats were observed under light microscope.The osteoclast was identified by tartrateresistant acid phosphatase (TRAP) staining.The calcineurin (CaN) activity of serum was measured by detection of free phosphate with malachite green.The bicinchoninic acid (BCA) method was used to detect total protein concentration of serum.The colorimetry method was used to detect calcium and malondialdehyde (MDA) levels in serum.The enzyme linked immunosorbent assay (ELISA) method was used to detect calmodulin (CaM) content.Results By the end of the experiment,none dental fluorosis was detected in control group,all rats in fluoride group had dental fluorosis.The bone fluoride content of rats in fluoride group [(4 460.671 ± 418.548) mg/kg] was about 7.6 times higher than that in control group [(582.534 ± 58.342) mg/kg,t =-29.020,P ＜ 0.01].Compared with the control group,the bone tissue of rats in fluoride group showed thicker bone trabecular,sclerotin fusion and incomplete mineralization.Positive signal intensity of TRAP staining of bone tissue in fluoride group was significantly higher than that in control group.The number of osteoclast formation in fluoride group [10 (5-12)] was significantly higher than that in control group [3 (2-4);U =92.5,P ＜ 0.01].CaN activity in serum of rats in fluoride group [(3.334 ± 0.654) nmol/mg prot] was significantly higher than that in control group [(1.289 ± 0.361) nmol/mg prot;t =-6.346,P ＜ 0.01].The Ca and CaM content of serum in rats were not significantly different between the two groups.However MDA content in fluoride group [(7.703 ± 2.954) μmol/L] was significantly higher than that in control group [(3.958 ± 1.965) μmol/L,t =-2.968,P ＜ 0.05].Conclusion Excessive fluoride may increase osteoclast formation in bone tissue of rats,and the mechanism might be fluoride stimulated CaN activity through oxidative stress pathway.

Dental stem cells (DSCs), an important source of mesenchymal stem cells (MSCs), can be easily obtained by minimally invasive procedures and have been used for the treatment of various diseases. Classic paradigm attributed the mechanism of their therapeutic action to direct cell differentiation after targeted migration, while contemporary insights into indirect paracrine effect opened new avenues for the mystery of their actual low engraftment and differentiation ability in vivo. As critical paracrine effectors, DSC-derived extracellular vesicles (DSC-EVs) are being increasingly linked to the positive effects of DSCs by an evolving body of in vivo studies. Carrying bioactive contents and presenting therapeutic potential in certain diseases, DSC-EVs have been introduced as promising treatments. Here, we systematically review the latest in vivo evidence that supports the therapeutic effects of DSC-EVs with mechanistic studies. In addition, current challenges and future directions for the clinical translation of DSC-EVs are also highlighted to call for more attentions to the (I) distinguishing features of DSC-EVs compared with other types of MSC-EVs, (II) heterogeneity among different subtypes of DSC-derived EVs, (III) action modes of DSC-EVs, (IV) standardization for eligible DSC-EVs and (V) safety guarantee for the clinical application of DSC-EVs. The present review would provide valuable insights into the emerging opportunities of DSC-EVs in future clinical applications.
Cell Differentiation ; Extracellular Vesicles/metabolism* ; Mesenchymal Stem Cell Transplantation/methods* ; Mesenchymal Stem Cells/metabolism*