• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Association Between Geranylgeranyl Pyrophosphate Synthase Gene Polymorphisms and Bone Phenotypes and Response to Alendronate Treatment in Chinese Osteoporotic Women△

    2016-09-06 02:21:15LanwenHanDoudouMaXiaojieXuFangYiLiuWeiboXiaYanJiangOuWangXiaopingXingandMeiLiDepartmentofEndocrinologyKeyLaboratoryofEndocrinologyofMinistryofHealthPekingUnionMedicalCollegeHospitalChineseAcademyofMedicalScienc
    Chinese Medical Sciences Journal 2016年1期

    Lan-wen Han, Dou-dou Ma, Xiao-jie Xu, Fang Lü, Yi Liu, Wei-bo Xia, Yan Jiang, Ou Wang, Xiao-ping Xing, and Mei Li*Department of Endocrinology, Key Laboratory of Endocrinology of Ministry of Health, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, 100730 Beijing, China

    ?

    Association Between Geranylgeranyl Pyrophosphate Synthase Gene Polymorphisms and Bone Phenotypes and Response to Alendronate Treatment in Chinese Osteoporotic Women△

    Lan-wen Han?, Dou-dou Ma?, Xiao-jie Xu?, Fang Lü, Yi Liu, Wei-bo Xia, Yan Jiang, Ou Wang, Xiao-ping Xing, and Mei Li*
    Department of Endocrinology, Key Laboratory of Endocrinology of Ministry of Health, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, 100730 Beijing, China

    geranylgeranyl pyrophosphate synthase; tag single nucleotide polymorphisms; osteoporosis; alendronate

    Objective To investigate the relationship between geranylgeranyl pyrophosphate synthase (GGPPS) gene polymorphisms and bone response to alendronate in Chinese osteoporotic women.

    Methods A total of 639 postmenopausal women with osteoporosis or osteopenia were included and randomly received treatment of low dose (70 mg per two weeks) or standard dose (70 mg weekly) of alendronate for one year. The six tag single nucleotide polymorphisms of GGPPS gene were identified. Bone mineral density (BMD), serum cross-linked C-telopeptide of type I collagen (β-CTX), and total alkaline phosphatase (ALP) were measured before and after treatment. GGPPS gene polymorphisms and the changes of BMD and bone turnover markers after treatment were analyzed.

    Results rs10925503 polymorphism of GGPPS gene was correlated to serum β-CTX levels at baseline, and patients with TT genotype had significantly higher serum β-CTX level than those with TC or CC genotype (all P<0.05). No correlation was found between polymorphisms of GGPPS gene and serum total ALP levels, as well as BMD at baseline. After 12 months of treatment, lumbar spine and hip BMD increased and serum bone turnover markers decreased significantly (P<0.01), and without obvious differences between the low dose and standard dose groups (all P>0.05). However, GGPPS gene polymorphisms were uncorrelated to percentage changes of BMD, serum total ALP, and β-CTX levels (all P>0.05).

    Conclusion GGPPS gene polymorphisms are correlated to osteoclasts activity, but all tag single nucleotide polymorphisms of GGPPS gene have no influence on the skeletal response to alendronate treatment.O STEOPOROSIS is a systemic disease characterized by impaired bone strength and increased bone fracture risk, which remarkably increases the morbidity and mortality of the elderly.1,2Osteoporosis is significantly influenced by genetic factors, and many genetic loci are identified to be associated with bone mineral density (BMD) and bone fracture risk.3,4Recently, bone responses to anti-osteoporotic agents are indicated to be influenced by genetic variance of encoding genes of vitamin D receptor,5estrogen receptor,6low density lipoprotein receptor-related protein,7,8farnesyl diphosphate synthase (FDPS),9and so on.

    Chin Med Sci J 2016; 31(1):8-16

    Bisphosphonates (BPs) are widely used as potent inhibitors of bone resorption, which significantly increase BMD and reduce bone fracture risk.10,11However, clinical studies indicate the response rate to BPs is variant from 62% to 90% among different individuals with osteoporosis.12-14Identification of the non-responders to BPs is beneficial to improve the treatment effects and decrease medical costs. Alendronate, the most commonly used nitrogen-containing BPs, can reduce osteoclast activity through inhibiting FDPS and geranylgeranyl pyrophosphate synthase (GGPPS) of mevalonate pathway.15,16Common allelic variants of FDPS gene have been demonstrated to influence the response of women with osteoporosis to BPs.9,17GGPPS is another important target of BPs in osteoclasts, which belongs to the trans-prenyltransferase family.18The enzyme catalyzes the synthesis of geranylgeranyl pyrophosphate (GGPP) from farnesyl diphosphate and isopentenyl diphosphate. GGPP is an important molecule responsible for the C20-prenylation of proteins and for the regulation of a nuclear hormone receptor.18GGPPS encoding gene has 5 exons and locates on chromosome 1q43.19As the crucial target of alendronate in the mevalonate pathway of osteoclasts, genetic variance of GGPPS gene is speculated to affect the skeletal response to alendronate. However, little is known about the influence of GGPPS gene polymorphisms on phenotype of osteoporosis and effects of alendronate. Therefore, we evaluate the influence of GGPPS gene polymorphisms on BMD, bone turnover biomarkers, and bone response to alendronate treatment in Chinese postmenopausal women with osteoporosis or osteopenia.

    PATIENTS AND METHODS

    Patients and treatment

    This is a multicenter, open-label, prospective study. During 2008 and 2011, more than 2000 postmenopausal women were screened from 7 clinical centers in China (Beijing, Changsha, Shanghai, Chengdu, Xi'an, Guangzhou, and Harbin). A total of 639 postmenopausal women with osteoporosis or osteopenia were enrolled in this study. Inclusion criteria were as follows: (1) patients aged 41-75 years old, with menopause age older than 40 years and years since menopause (YSM) more than 1 year; (2) T-score of BMD at the lumbar spine or femoral neck less than -1; (3) at least 3 lumbar vertebras could be measured; (4) could ambulate at the least 30 minutes in each day. Exclusion criteria were as follows: (1) patients with severe liver or kidney diseases; (2) with severe gastrointestinal diseases; (3) intolerance to BPs; (4) with treatment history of BPs or parathyroid hormone 1-34 within recent 12 months; with treatment of estrogen, selective estrogen receptor modulators, active vitamin D analogues or calcitonin within recent 6 months; (5) with more than 2 weeks therapy history of corticosteroid or anticonvulsant therapy within recent 3 months; (6) with other metabolic or inherited bone diseases; (7) with autoimmune diseases.

    Postmenopausal women with osteoporosis or osteopenia randomly received low-dose (70 mg every two weeks) or standard-dose (70 mg weekly) of alendronate (Shijiazhuang Ouyi Pharmaceutical Co. Ltd., Hebei, China) for 1 year. Patients took alendronate on an empty stomach at least 30 minutes before breakfast with 250 ml plain water, and remain upright for at least 30 minutes after dosing. All patients were supplemented with 600 mg elemental calcium plus 125 IU of vitamin D3daily (Caltrate D; Pfizer Inc., USA). Compliance to alendronate treatment was calculated according to the number of remaining tablets during the follow-up.

    The study was approved by the Ethical Committee of Peking Union Medical College Hospital and all participants signed informed consents.

    Genotyping

    Genomic DNA was extracted from leukocytes of peripheral blood according to standard procedures. The Entrez Gene Database (http://www.ncbi.nlm.nih.gov/gene/) and HapMap (http://hapmap.ncbi.nlm.nih.gov/) were adopted to identify single nucleotide polymorphisms (SNPs) information of GGPPS gene in Chinese population. Six tag SNPs of GGPPS were selected as follows: rs2803851, rs2789367, rs10802624, rs10925503, rs3840452, and rs2789366, which could almost represent the whole genetic variance of this gene. The genotypes of rs3840452 of GGPPS gene were assayed by short tandem repeat. The genotypes of GGPPS gene of rs2803851, rs2789367, rs10802624, rs10925503, and rs2789366 were detected by TaqMan allelic discrimination assay (Applied Biosystems, USA). Primer sequences and the distributions of tag SNPs of GGPPS gene are shown inTable 1. The whole reacting volume was 6 μl, including 3 μl DNA sample, 2.5 μl TaqMan Universal PCR Master Mix (Applied Biosystems, USA), 0.125 μl TaqMan probe assay (including primers), and 0.375 μl ddH2O. Reactions were performed on a Real-time PCR system of ABI Prism7900 (Applied Biosystems) under standard condition.

    Evaluation of bone response to alendronate

    Skeletal response to alendronate was assessed by percentage changes of BMD and bone turnover biomarkers after treatment. BMD at the lumbar spine and proximal femur was measured by dual-energy X-ray absorptiometry (DXA) (Hologic or Lunar) at baseline and after 12 months of treatment. Cross calibration equations between two kinds of machine were as follows: Hologic BMD (g/cm2) = 0.802 –Lunar + 0.318 (r=0.991, P<0.001, SE=0.03 g/cm2).20BMD phantom scan was measured by each DXA instrument daily and no significant machine drift was detected during the whole period of study. The coefficients of variation (CV) of BMD were 0.8%-1.0% for the lumbar spine and total hip. Radiologists were responsible for the analysis of BMD data, quality control, and phantom scan, who were unaware of the study group assignments. According to the criteria of World Health Organization, osteoporosis was defined as BMD T-score ≤ -2.5 at the lumbar spine, femoral neck or total hip, and -2.5 < T-score ≤ -1.0 at above sites was considered as osteopenia.21Participants were defined as severe osteoporosis if they had fragility fracture history.

    Serum calcium, phosphate, total alkaline phosphatase (total ALP, as a bone formation marker), cross-linked C-telopeptide of type I collagen (β-CTX, as a bone resorption marker) and 25 hydroxy-vitamin D (25OHD, as a marker of vitamin D nutritional status) levels were measured at baseline and after 12 months of treatment. Serum β-CTX and 25OHD levels were detected by electrochemiluminescence immunoassay (Roche Diagnostics Co. Ltd., Germany). The lowest detection limit of β-CTX and 25OHD was 0.01 ng/ml and 4.0 ng/ml, respectively. CV of intra-assay and inter-assay of β-CTX were 2.2%-4.6% and 2.5%-4.7%, respectively. The intra-assay and inter-assay CV of 25OHD were 4.1%-5.7% and 6.6%-9.9%, respectively.

    Table 1. Primer sequences of Tag SNPs of GGPPS gene

    Statistical analysis

    Data of normal distribution were presented as mean ± standard deviation (SD), while those of abnormal distribution were expressed as median and quartiles. Genotype distributions of GGPPS gene were tested for Hardy-Weinberg using Chi-square test. Associations between genotypes of GGPPS gene and BMD, as well as serum levels of calcium, phosphate, total ALP, β-CTX, 25OHD were evaluated using analysis of variance adjusted for age, body mass index (BMI), and YSM. The response to alendronate treatment was estimated as the percentage changes of BMD and serum bone turnover biomarkers (total ALP and β-CTX), which were calculated with the following formula: (parameters after treatment-parameters at baseline)/ parameters at baseline × 100%. Association of the genotypes and percentage changes of BMD, total ALP, β-CTX after treatment was evaluated by analysis of variance and multiple linear regression adjusted for age, BMI, and YSM. The statistical analyses were performed using SPSS 17.0 (SPSS Inc., Chicago, IL, USA). Statistical significance was considered when P value was less than 0.05.

    RESULTS

    Characteristics of study population at baseline

    Demographic characteristics of the postmenopausal women with osteoporosis or osteopenia are shown in Table 2. No significant differences were found in age, YSM, BMI, BMD, serum 25OHD, total ALP, and β-CTX levels between the low dose and standard dose groups(all P>0.05).

    GGPPS gene polymorphisms and bone phenotypes

    Genotypes and allele distributions of the six tag SNPs of GGPPS gene are shown in Table 3. All SNPs were in Hardy-Weinberg equilibrium. At baseline, rs10925503 polymorphism of GGPPS gene was correlated to serum β-CTX levels, which were 0.45±0.02, 0.41±0.02, and 0.36±0.02 ng/ml in women with TT, TC, and CC genotypes, after adjusting for age, YSM, and BMI (all P<0.05). The other SNPs of GGPPS gene were uncorrelated to serum β-CTX levels (Fig. 1A). The serum bone formation marker (total ALP)levels did not present significant difference among genotypes of all tag SNPs of GGPPS gene (all P>0.05) (Fig. 1B). BMD at the lumbar spine and femoral neck had no obvious difference among all genotypes of GGPPS gene, after adjusting for age, YSM, and BMI (all P>0.05, Fig. 1C and 1D).

    Patients with TT genotype of rs10925503 polymorphism of GGPPS gene had significantly higher serum β-CTX level than those with TC or CC genotype (P=0.012). The serum ALP levels, BMD at the lumbar spine and femoral neck did not present significant difference among genotypes of all tag SNPs of GGPPS (all P>0.05).

    GGPPS gene polymorphisms and skeletal response to alendronate treatment

    There were 540 (84.5%) women who completed the 12 months of treatment. And, 99 (15.5%) women withdrew from the treatment because of clinical adverse effects, protocol deviation, losing to follow-up and other reasons. After 12 months of treatment, no differences was found in BMD changes at the lumbar spine, femoral neck, and total hip between the standard-dose group (n=266; 5.07%, 2.93%, and 3.80%, respectively) and the low-dose group (n=274; 5.60%, 3.87%, and 3.28%, respectively; all P>0.05). In the low-dose and standard-dose groups, the serum levels of total ALP decreased by 24.6% and 29.1%, and serum β-CTX levels decreased dramatically by 57.0% and 69.7% after 12 months of treatment. However, after low dose or standard dose treatment of alendronate, the percentage changes of BMD at the lumbar spine and proximal femur presented no significant difference among different genotypes of GGPPS after adjusting for age, YSM, and BMI (Tables 4 and 5). No significant differences in percentage changes of serum ALP and β-CTX levels were found among different genotypes of GGPPS gene (Tables 4 and 5).

    Table 2. Demographic characteristics of the study population at baseline§

    Table 3. Genotypes and allele distributions of the six tag SNPs of GGPPS gene

    Figure 1. Baseline serum β-CTX (A) and ALP levels (B), BMD at the lumbar spine (C) and femoral neck (D) among different genotypes of tag SNPs of GGPPS gene.*P<0.05 compared with women with TT genotype.

    Table 4. The percentage change of BMD and bone turnover biomarkers among different genotypes after 12 months of low-dose of alendronate treatment (%)

    Table 5. The percentage changes of BMD and bone turnover biomarkers among different genotypes after 12 months of standard-dose of alendronate treatment (%)

    DISCUSSION

    Osteoporosis has a strong hereditary background and represents an important health problem among the elderly. Nitrogen-containing BPs are widely used in the treatment of osteoporosis and diseases with high bone remodeling, which have been demonstrated to reduce the production of farnesyl pyrophosphate (FPP) and GGPP through inhibition of crucial enzymes FDPS and GGPPS in the mevalonate pathway of osteoclasts.22,23The mechanism of alendronate suppression of osteoclast formation is also directly correlated to the inhibition of GGPPS.24-26Therefore, it is important to evaluate the correlation of genetic variation in GGPPS and bone phenotypes, as well as skeletal responsiveness to alendronate. However, the pharmacogenetic information on GGPPS and alendronate treatment is still scanty.

    In this study, we found that the frequencies of GGPPS polymorphisms of rs3840452 in Chinese women were similar to those in Korean,27and genotypes frequencies of rs2803851, rs2789367, rs10802624, rs10925503 of GGPPS in Chinese women were different from those in Japanese, European and African according to data in http://hapmap. ncbi.nlm.nih.gov/. We only found rs10925503 polymorphism of GGPPS was correlated to baseline level of bone resorption marker, and patients with TT genotype had significantly higher serum β-CTX level than those with TC or CC genotype. The location of rs10925503 is close to 3’ untranslated region of GGPPS, a possible binding site of transcription factor, which could regulate expression of GGPPS. Therefore, rs10925503 polymorphism of GGPPS was correlated to the activity of osteoclasts. No association was found between GGPPS polymorphisms with bone formation marker and BMD at baseline. In a small sample of Korean women, -/-genotype of GGPPS rs3840452 was found to be correlated to higher femoral neck BMD than AA or A/- genotypes,27which was inconsistent with our results. As there were only 7 Korean women with -/- genotype, a bias of the small sample in that study could not be ruled out.

    As we know, according to changes of BMD, it was estimated that about 5%-10% of patients do not respond to anti-osteoporotic therapy.28Sequence variants in the human genome are important causes of difference in drug responses. Pharmacogenetic study in osteoporosis contributes to improve drug efficacy and safety through identification of genetic markers of different patients. Several studies indicated that GGPPS was the crucial target of BPs in mevalonate pathway of osteoclasts. BPs effectively blocked protein geranylgeranylation through inhibition of GGPPS, which led to ultrastructural changes of osteoclasts at lower concentrations and to apoptosis at high concentrations.29,30It would be of important clinical value to determine the individualized anti-osteoporosis therapy on the basis of the pharmacogenetic information of GGPPS and BPs. We detected all tag SNPs of GGPPS, which could almost represent the whole genetic variance of this gene. We evaluate the correlation between the genetic variation of GGPPS and bone response to alendronate in large sample of Chinese women. However, according to percentage changes of BMD and bone turnover biomarkers, no correlation was found between the genetic variation of GGPPS and bone responsiveness to alendronate, either in low-dose or in standard-dose group. Consistently, in Korean women, GGPPS rs3840452 polymorphism was also uncorrelated to the effects of alendronate.27

    Our study has several limitations. Most of the ALP isoenzymes are derived from the bones and liver. Total ALP was measured rather than bone specific ALP in our study. The alendronate treatment period was rather short. We found serum 25OHD level was very low in this population, which was reported to affect the response to anti-resorbing drugs.31All patients were supplemented with only 125 IU of vitamin D3, which could not rule out the influence of vitamin D deficiency on the response to alendronate therapy. On the other hand, we found the correlation of rs10925503 polymorphism of GGPPS with β-CTX level, but the mechanisms involved in the genotype-related differences remained to be elucidated. We did not investigate the correlation between the GGPPS polymorphisms and other kinds of aminobisphosphonate, such as zoledronic acid or ibandronate, so the results could not represent other BPs and GGPPS.

    In conclusion, rs10925503 polymorphism of GGPPS gene is correlated to activity of osteoclasts, but all tag SNPs of GGPPS have no influence on baseline BMD and skeletal response to alendronate. Therefore, GGPPS could only act as a candidate gene for bone resorption, instead of a pharmacogenetic gene for forecasting effects of alendronate.

    ACKNOWLEDGMENTS

    All the samples in the study come from seven clinical centers in China (Beijing, Changsha, Shanghai, Chengdu, Xi’an, Guangzhou, and Harbin). We acknowledge the support of clinical data collection for the research.

    REFERENCES

    1. Prior JC, Langsetmo L, Lentle BC, et al. Ten-year incident osteoporosis-related fractures in the population-based Canadian multicentre osteoporosis study?Comparing site and age-specific risks in women and men. Bone 2015;71:237-43.

    2. Berglundh S, Malmgren L, Luthman H, et al. C-reactive protein, bone loss, fracture, and mortality in elderly women: a longitudinal study in the OPRA cohort. Osteoporos Int 2015; 26:727-35.

    3. Mitchell BD, Streeten EA. Clinical impact of recent genetic discoveries in osteoporosis. Appl Clin Genet 2013; 6: 75-85.

    4. Lee SH, Lee SW, Ahn SH, et al. Multiple gene polymorphisms can improve prediction of nonvertebral fracture in postmenopausal women. J Bone Miner Res 2013; 28: 2156-64.

    5. Otrock ZK, Mahfouz RA, Charafeddine KM, et al. Vitamin D receptor genotypes and response to zoledronic acid therapy in thalassemia-induced osteoporosis. Ann Hematol 2008; 87:947-8.

    6. Sai AJ, Gallagher JC, Fang X. Effect of hormone therapy and calcitriol on serum lipid profile in postmenopausal older women: association with estrogen receptor-α genotypes. Menopause 2011; 18:1101-12.

    7. Kruk M, Ralston SH, Albagha OM. LRP5 polymorphisms and response to risedronate treatment in osteoporotic men. Calcif Tissue Int 2009; 84:171-9.

    8. Zhou PR, Liu HJ, Liao EY, et al. LRP5 polymorphisms and response to alendronate treatment in Chinese postmenopausal women with osteoporosis. Pharmacogenomics 2014; 15:821-31.

    9. Olmos JM, Zarrabeitia MT, Hernández JL, et al. Common allelic variants of the farnesyl diphosphate synthase gene influence the response of osteoporotic women to bisphosphonates. Pharmacogenomics J 2012; 12:227-32.

    10. Favus MJ. Bisphosphonates for osteoporosis. NEJM 2010; 363:2027-35.

    11. Russell RG. Bisphosphonates: the first 40 years. Bone 2011; 49:2-19.

    12. Burnett-Bowie SM, Saag K, Sebba A. Prediction of changes in bone mineral density in postmenopausal women treated with once-weekly bisphosphonates. J Clin Endocrinol Metab 2009; 94:1097-103.

    13. Emkey R, Delmas PD, Bolognese M, et al. Efficacy and tolerability of once-monthly oral ibandronate (150 mg) and once-weekly oral alendronate (70 mg): additional results from the monthly oral therapy with ibandronate for osteoporosis intervention (MOTION) study. Clin Ther 2009; 31:751-61.

    14. Orwoll ES1, Binkley NC, Lewiecki EM. Efficacy and safety of monthly ibandronate in men with low bone density. Bone 2010; 46:970-6.

    15. Tsubaki M, Komai M, Itoh T, et al. Nitrogen-containing bisphosphonates inhibit RANKL- and M-CSF-induced osteoclast formation through the inhibition of ERK1/2 and Akt activation. J Biomed Sci 2014; 21:10.

    16. Dudakovic A, Wiemer AJ, Lamb KM, et al. Inhibition of geranylgeranyl pyrophosphate synthase induces apoptosis through multiple mechanisms and displays synergy with inhibition of other isoprenoid biosynthetic enzyme. JPET 2008; 324:1028-36.

    17. Liu Y, Liu H, Li M, et al. Association of farnesyl diphosphate synthase polymorphisms and response to alendronate treatment in Chinese postmenopausal women with osteoporosis. Chin Med J 2014; 127:662-8.

    18. Chen SH, Lin SW, Lin SR, et al. Moiety-linkage map reveals selective nonbisphosphonate inhibitors of human geranylgeranyl diphosphate synthase. J Chem Inf Model 2013; 53:2299-311.

    19. Ericsson J, Greene JM, Carter KC, et al. Human geranylgeranyl diphosphate synthase: isolation of the cDNA, chromosomal mapping and tissue expression. J Lipid Res 1998; 39:1731-9.

    20. Cummings SR, Bates D, Black DM. Clinical use of bone densitometry scientific review. JAMA 2002; 288:1889-97.

    21. Kanis JA, Melton LJ, Christiansen C, et al. The diagnosis of osteoporosis. J Bone Miner Res 1994; 9:1137-41.

    22. Kavanagh KL, Guo K, Dunford JE, et al. The molecular mechanism of nitrogen-containing bisphosphonates as antiosteoporosis drugs. Proc Natl Acad Sci USA 2006; 103:7829-34.

    23. Tsubaki M, Kato C, Nishinobo M, et al. Nitrogen-containing bisphosphonate, YM529/ONO-5920, inhibits macrophage inflammatory protein 1 alpha expression and secretion in mouse myeloma cells. Cancer Sci 2008; 99:152-8.

    24. Ling Y, Li ZH, Miranda K, et al. The farnesyl-dipho-sphate/ geranylgeranyl-diphosphate synthase of toxoplasma gondii is a bifunctional enzyme and a molecular target of bisphosphonates. J Biol Chem 2007; 282:30804-16.

    25. Fisher JE, Rogers MJ, Halasy JM, et al. Alendronate mechanism of action: geranylgeraniol, an intermediate in the mevalonate pathway, prevents inhibition of osteoclast formation, bone resorption, and kinase activation in vitro. Proc Natl Acad Sci USA 1999; 96:133-8.

    26. Coxon FP, Helfrich MH, Van’t Hof R, et al. Protein geranylgeranylation is required for osteoclast formation, function, and survival: inhibition by bisphosphonates and GGTI-298. J Bone Miner Res 2000; 15:1467-76.

    27. Choi HJ, Choi JY, Cho SW. Genetic polymorphism of geranylgeranyl diphosphate synthase (GGSP1) predicts bone density response to bisphosphonate therapy in Korean women. Yonsei Med J 2010; 51:231-8.

    28. Marini F, Brandi ML. Pharmacogenetics of osteoporosis: future perspectives. Calcif Tissue Int 2009; 84:337-47.

    29. Benford, HL, McGowan NW, Helfrich MH, et al. Visualization of bisphosphonate-induced caspase-3 activity in apoptotic osteoclasts in vitro. Bone 2001; 28:465-73.

    30. Halasy-Nagy JM, Rodan GA, Reszka AA. Inhibition of bone resorption by alendronate and risedronate does not require osteoclast apotosis. Bone 2001; 29:553-9.

    31. Ishijima M, Sakamoto Y, Yamanaka M, et al. Minimum required vitamin D level for optimal increase in bone mineral density with alendronate treatment in osteoporotic women. Calcif Tissue Int 2009; 85:398-404.

    for publication May 24, 2015.

    ?These authors contributed equally to this work.

    *Corresponding author Tel: 86-10-69155088, Fax: 86-10-69155088, E-mail: limeilzh@sina.com

    △Supported by National Natural Science Foundation of China (81570802) and National Key Program of Clinical Science (WBYZ2011-873).

    亚洲美女黄色视频免费看| 国国产精品蜜臀av免费| 精品国产乱码久久久久久小说| 国产综合精华液| 久久国产乱子免费精品| 国产一区二区三区综合在线观看 | 天堂中文最新版在线下载| 亚洲成人一二三区av| 亚洲精品久久午夜乱码| 国产欧美日韩综合在线一区二区 | 欧美三级亚洲精品| 日韩精品免费视频一区二区三区 | 欧美 日韩 精品 国产| 免费观看性生交大片5| 国产伦精品一区二区三区视频9| 久久婷婷青草| 韩国av在线不卡| 亚洲综合精品二区| 日本av手机在线免费观看| 日韩中文字幕视频在线看片| 搡女人真爽免费视频火全软件| 国产深夜福利视频在线观看| 校园人妻丝袜中文字幕| 久久久久人妻精品一区果冻| 午夜久久久在线观看| 国产熟女午夜一区二区三区 | 国产一区有黄有色的免费视频| av国产久精品久网站免费入址| 国产成人91sexporn| 中文欧美无线码| 男的添女的下面高潮视频| 秋霞伦理黄片| 国产精品一二三区在线看| 天美传媒精品一区二区| 99久久精品热视频| 色94色欧美一区二区| 久久精品久久久久久久性| 国产精品久久久久久精品电影小说| av卡一久久| 综合色丁香网| 天堂8中文在线网| 国产精品三级大全| 九九在线视频观看精品| 亚洲av在线观看美女高潮| 亚洲,一卡二卡三卡| 亚洲国产精品一区三区| 一本色道久久久久久精品综合| 一级,二级,三级黄色视频| 国产av精品麻豆| 五月开心婷婷网| 久久久久久久亚洲中文字幕| 精品久久久噜噜| 99热这里只有是精品50| 黑人巨大精品欧美一区二区蜜桃 | 乱人伦中国视频| 色婷婷久久久亚洲欧美| 校园人妻丝袜中文字幕| 中文精品一卡2卡3卡4更新| 久久久久久伊人网av| 亚洲伊人久久精品综合| 交换朋友夫妻互换小说| 精品酒店卫生间| 国产成人午夜福利电影在线观看| 极品教师在线视频| 少妇人妻精品综合一区二区| 一本大道久久a久久精品| 一本大道久久a久久精品| 乱系列少妇在线播放| 人妻系列 视频| 国产精品伦人一区二区| 成年av动漫网址| 日韩免费高清中文字幕av| 国产男人的电影天堂91| 18禁裸乳无遮挡动漫免费视频| 久久99蜜桃精品久久| 女性生殖器流出的白浆| 亚洲精华国产精华液的使用体验| 亚洲av不卡在线观看| 亚洲欧美日韩另类电影网站| 亚洲av免费高清在线观看| 国模一区二区三区四区视频| 国产亚洲欧美精品永久| 国产极品天堂在线| 亚洲中文av在线| 久久久精品免费免费高清| 亚洲不卡免费看| 免费黄色在线免费观看| 卡戴珊不雅视频在线播放| 久久久精品94久久精品| 日本vs欧美在线观看视频 | 午夜免费鲁丝| 国产国拍精品亚洲av在线观看| 在线观看免费日韩欧美大片 | 日韩在线高清观看一区二区三区| 内地一区二区视频在线| 97精品久久久久久久久久精品| 一级二级三级毛片免费看| 国产一区亚洲一区在线观看| av播播在线观看一区| 永久免费av网站大全| 亚洲经典国产精华液单| 日日摸夜夜添夜夜添av毛片| 亚洲国产毛片av蜜桃av| 精品人妻一区二区三区麻豆| 男人和女人高潮做爰伦理| 国产免费一区二区三区四区乱码| 亚洲精品国产色婷婷电影| 丰满乱子伦码专区| 18禁在线播放成人免费| 一本久久精品| 国产精品一二三区在线看| 国产亚洲精品久久久com| 亚洲伊人久久精品综合| 久久久久久伊人网av| 中文精品一卡2卡3卡4更新| 久久影院123| 一级a做视频免费观看| 91久久精品国产一区二区三区| 久久久精品94久久精品| 亚洲伊人久久精品综合| 2018国产大陆天天弄谢| 少妇裸体淫交视频免费看高清| 少妇裸体淫交视频免费看高清| av卡一久久| 高清欧美精品videossex| 美女xxoo啪啪120秒动态图| 午夜久久久在线观看| 日韩中文字幕视频在线看片| 一区二区av电影网| 欧美bdsm另类| 亚洲精品,欧美精品| 男的添女的下面高潮视频| 永久网站在线| 国产成人aa在线观看| 午夜久久久在线观看| 国产精品人妻久久久影院| 国产伦精品一区二区三区视频9| 久久99精品国语久久久| 22中文网久久字幕| 边亲边吃奶的免费视频| 夜夜看夜夜爽夜夜摸| 成人亚洲欧美一区二区av| 大香蕉久久网| 伊人久久精品亚洲午夜| 色5月婷婷丁香| 午夜免费男女啪啪视频观看| 国产淫语在线视频| 又黄又爽又刺激的免费视频.| 在线亚洲精品国产二区图片欧美 | 国精品久久久久久国模美| 国产av国产精品国产| 免费观看性生交大片5| 六月丁香七月| 六月丁香七月| 欧美xxxx性猛交bbbb| 一级av片app| 久久精品国产a三级三级三级| 中文字幕人妻熟人妻熟丝袜美| 精品亚洲成a人片在线观看| 九九爱精品视频在线观看| 内地一区二区视频在线| 亚洲第一av免费看| 国产欧美亚洲国产| 成人毛片a级毛片在线播放| 蜜臀久久99精品久久宅男| a级毛片在线看网站| 汤姆久久久久久久影院中文字幕| 男的添女的下面高潮视频| 天天操日日干夜夜撸| 大话2 男鬼变身卡| 中文字幕人妻丝袜制服| 一级,二级,三级黄色视频| 伊人亚洲综合成人网| 天天躁夜夜躁狠狠久久av| 国产精品一区二区性色av| 色94色欧美一区二区| 国产高清不卡午夜福利| 人体艺术视频欧美日本| 国产白丝娇喘喷水9色精品| 看免费成人av毛片| 日韩熟女老妇一区二区性免费视频| 少妇精品久久久久久久| 亚洲av成人精品一区久久| 亚洲精品一区蜜桃| 久久久国产欧美日韩av| 伊人久久国产一区二区| 亚洲av在线观看美女高潮| 乱码一卡2卡4卡精品| 日本91视频免费播放| 国产色爽女视频免费观看| 日韩视频在线欧美| 久久这里有精品视频免费| 99久久精品一区二区三区| 在线观看一区二区三区激情| 国产一区有黄有色的免费视频| 久久久精品免费免费高清| 久久久国产精品麻豆| 日日啪夜夜撸| 日韩不卡一区二区三区视频在线| 色吧在线观看| 夫妻午夜视频| 亚洲欧洲精品一区二区精品久久久 | 在线观看免费视频网站a站| av专区在线播放| 日本91视频免费播放| a级一级毛片免费在线观看| 欧美最新免费一区二区三区| 日韩,欧美,国产一区二区三区| 国产高清不卡午夜福利| 日本av免费视频播放| 亚洲av福利一区| 人人澡人人妻人| 美女xxoo啪啪120秒动态图| 综合色丁香网| 日日爽夜夜爽网站| 一本大道久久a久久精品| 曰老女人黄片| 久久精品国产亚洲网站| 久久久久久久大尺度免费视频| 国产美女午夜福利| a级毛色黄片| 久久久久久久国产电影| 老女人水多毛片| h视频一区二区三区| 国产精品99久久久久久久久| av有码第一页| 亚洲欧美一区二区三区黑人 | 久久久久久久久久成人| 亚洲成色77777| 欧美成人午夜免费资源| 乱码一卡2卡4卡精品| 欧美老熟妇乱子伦牲交| 欧美最新免费一区二区三区| 日日啪夜夜撸| 午夜久久久在线观看| 91久久精品国产一区二区成人| 久久97久久精品| 精品卡一卡二卡四卡免费| a级毛色黄片| 日韩一区二区视频免费看| 久久国产精品大桥未久av | 成人特级av手机在线观看| 免费大片18禁| 国产黄色视频一区二区在线观看| 日韩一区二区三区影片| 纯流量卡能插随身wifi吗| 人妻 亚洲 视频| 亚洲国产日韩一区二区| .国产精品久久| 日韩一本色道免费dvd| 我的女老师完整版在线观看| 国产亚洲午夜精品一区二区久久| 国产精品人妻久久久影院| 亚洲欧洲精品一区二区精品久久久 | 婷婷色综合大香蕉| 菩萨蛮人人尽说江南好唐韦庄| 我的女老师完整版在线观看| √禁漫天堂资源中文www| 嫩草影院新地址| 丰满少妇做爰视频| 最近的中文字幕免费完整| 91精品国产九色| 一个人免费看片子| 日韩在线高清观看一区二区三区| 国产伦在线观看视频一区| 欧美激情极品国产一区二区三区 | 午夜免费鲁丝| 五月天丁香电影| 99热这里只有精品一区| 国产成人一区二区在线| a级毛片在线看网站| a级毛片在线看网站| 街头女战士在线观看网站| 亚州av有码| 成人18禁高潮啪啪吃奶动态图 | 99热6这里只有精品| 国产日韩欧美亚洲二区| 性高湖久久久久久久久免费观看| 人人妻人人澡人人爽人人夜夜| 看十八女毛片水多多多| 欧美精品一区二区大全| 亚洲精品色激情综合| 亚洲情色 制服丝袜| 欧美xxxx性猛交bbbb| 欧美一级a爱片免费观看看| 国产男女超爽视频在线观看| 欧美日韩视频精品一区| 免费不卡的大黄色大毛片视频在线观看| 色婷婷av一区二区三区视频| 丰满乱子伦码专区| 99久久综合免费| 免费在线观看成人毛片| 精品亚洲成a人片在线观看| 国产伦理片在线播放av一区| 精品国产一区二区三区久久久樱花| 我的老师免费观看完整版| 国产一区二区三区av在线| 久久久久视频综合| 久久午夜福利片| av黄色大香蕉| 国产高清不卡午夜福利| 最黄视频免费看| 一区二区三区四区激情视频| 成人二区视频| 人人妻人人爽人人添夜夜欢视频 | 一级,二级,三级黄色视频| 国产真实伦视频高清在线观看| 一边亲一边摸免费视频| 一本大道久久a久久精品| 精品亚洲乱码少妇综合久久| 两个人免费观看高清视频 | 欧美国产精品一级二级三级 | 亚洲第一av免费看| 大香蕉久久网| 深夜a级毛片| 美女xxoo啪啪120秒动态图| 99热全是精品| 亚洲国产毛片av蜜桃av| 亚洲国产日韩一区二区| 婷婷色麻豆天堂久久| 日韩大片免费观看网站| 色视频在线一区二区三区| 下体分泌物呈黄色| 久久精品国产亚洲av天美| 色5月婷婷丁香| 22中文网久久字幕| 18禁裸乳无遮挡动漫免费视频| 狂野欧美激情性xxxx在线观看| 在线观看免费高清a一片| 欧美xxxx性猛交bbbb| 又黄又爽又刺激的免费视频.| 亚洲无线观看免费| 亚洲中文av在线| 街头女战士在线观看网站| 国产亚洲午夜精品一区二区久久| 嫩草影院入口| 免费看日本二区| 少妇的逼水好多| 纵有疾风起免费观看全集完整版| 国产欧美日韩综合在线一区二区 | 美女内射精品一级片tv| 在线观看www视频免费| 久久久精品免费免费高清| 天堂中文最新版在线下载| 日本wwww免费看| 一区二区三区四区激情视频| 99热这里只有是精品在线观看| 一个人免费看片子| 夫妻性生交免费视频一级片| 亚洲欧美成人综合另类久久久| 欧美性感艳星| 国产精品国产三级国产专区5o| 色5月婷婷丁香| 美女国产视频在线观看| 亚洲精品成人av观看孕妇| 尾随美女入室| 久久99蜜桃精品久久| 国产成人免费无遮挡视频| 国产在线一区二区三区精| 久久人妻熟女aⅴ| 黑人猛操日本美女一级片| 麻豆乱淫一区二区| 五月开心婷婷网| 18禁裸乳无遮挡动漫免费视频| videossex国产| 日本91视频免费播放| .国产精品久久| 又大又黄又爽视频免费| 一区二区三区四区激情视频| 亚洲精品一区蜜桃| 久久99一区二区三区| 国产视频首页在线观看| 女的被弄到高潮叫床怎么办| 欧美变态另类bdsm刘玥| 亚洲av国产av综合av卡| 综合色丁香网| 国产亚洲av片在线观看秒播厂| 欧美最新免费一区二区三区| av福利片在线| 王馨瑶露胸无遮挡在线观看| 国产精品麻豆人妻色哟哟久久| 国内少妇人妻偷人精品xxx网站| 97在线视频观看| 亚洲精品中文字幕在线视频 | 人人妻人人爽人人添夜夜欢视频 | 亚洲国产精品专区欧美| 一区二区三区免费毛片| 国产深夜福利视频在线观看| 女人久久www免费人成看片| 免费播放大片免费观看视频在线观看| 国产精品女同一区二区软件| 色视频www国产| 麻豆成人av视频| 亚洲天堂av无毛| 久久6这里有精品| 亚洲精品乱码久久久v下载方式| 伊人久久精品亚洲午夜| av免费观看日本| 卡戴珊不雅视频在线播放| 亚洲精品乱码久久久v下载方式| 国产在线视频一区二区| 国内揄拍国产精品人妻在线| 午夜福利视频精品| 熟妇人妻不卡中文字幕| av天堂中文字幕网| xxx大片免费视频| 亚洲av综合色区一区| 九九在线视频观看精品| 一级毛片电影观看| 内地一区二区视频在线| 国产免费一级a男人的天堂| 久久人人爽av亚洲精品天堂| 内地一区二区视频在线| 高清黄色对白视频在线免费看 | 亚洲精品456在线播放app| 国产精品.久久久| 亚洲精品色激情综合| 青春草国产在线视频| 久久女婷五月综合色啪小说| 多毛熟女@视频| 国产爽快片一区二区三区| 国产白丝娇喘喷水9色精品| 亚洲综合色惰| 亚洲精品第二区| 久久久久久久久久人人人人人人| 日本wwww免费看| 最近手机中文字幕大全| 免费看不卡的av| 精品一区二区免费观看| 熟女人妻精品中文字幕| 国产在线视频一区二区| 中文精品一卡2卡3卡4更新| 免费看日本二区| 夫妻性生交免费视频一级片| 国产一区亚洲一区在线观看| 51国产日韩欧美| 中文资源天堂在线| 五月伊人婷婷丁香| 丰满少妇做爰视频| 国产av国产精品国产| 国产日韩欧美视频二区| 波野结衣二区三区在线| 国国产精品蜜臀av免费| 欧美成人午夜免费资源| 最新的欧美精品一区二区| 一级毛片我不卡| 久久精品国产亚洲av涩爱| 国产亚洲欧美精品永久| 一级av片app| 久久久亚洲精品成人影院| 91久久精品国产一区二区成人| 老熟女久久久| 亚洲色图综合在线观看| 熟妇人妻不卡中文字幕| 这个男人来自地球电影免费观看 | 妹子高潮喷水视频| 日韩,欧美,国产一区二区三区| 亚洲国产精品专区欧美| 建设人人有责人人尽责人人享有的| 97超碰精品成人国产| 3wmmmm亚洲av在线观看| 高清欧美精品videossex| 亚洲欧美日韩东京热| 日韩av免费高清视频| 久热久热在线精品观看| 男女啪啪激烈高潮av片| 人人澡人人妻人| 丝袜喷水一区| 日韩av在线免费看完整版不卡| 国产高清国产精品国产三级| 自线自在国产av| 久久热精品热| 亚洲精品久久午夜乱码| av在线老鸭窝| 亚洲熟女精品中文字幕| 久久精品国产亚洲网站| 永久免费av网站大全| av不卡在线播放| 国产又色又爽无遮挡免| 亚洲美女视频黄频| 黄色视频在线播放观看不卡| 欧美日本中文国产一区发布| 爱豆传媒免费全集在线观看| 丰满饥渴人妻一区二区三| .国产精品久久| 精品一区在线观看国产| 在线亚洲精品国产二区图片欧美 | 97超碰精品成人国产| 国产一区二区在线观看av| 狂野欧美激情性bbbbbb| 黄色日韩在线| 日本猛色少妇xxxxx猛交久久| 性色av一级| 欧美三级亚洲精品| 校园人妻丝袜中文字幕| 黄色欧美视频在线观看| 免费观看a级毛片全部| 春色校园在线视频观看| 亚洲,欧美,日韩| 日韩中文字幕视频在线看片| 国产69精品久久久久777片| 欧美三级亚洲精品| 校园人妻丝袜中文字幕| 久久狼人影院| 国产一区二区在线观看日韩| 五月天丁香电影| 国产精品.久久久| av一本久久久久| 丁香六月天网| av福利片在线| 热re99久久精品国产66热6| 寂寞人妻少妇视频99o| 亚洲,一卡二卡三卡| 午夜免费男女啪啪视频观看| 搡女人真爽免费视频火全软件| 最近中文字幕高清免费大全6| 免费看日本二区| 久久久亚洲精品成人影院| 午夜91福利影院| 亚洲,欧美,日韩| 久久久久久伊人网av| 极品教师在线视频| 日韩欧美 国产精品| 91在线精品国自产拍蜜月| 精品亚洲成国产av| 哪个播放器可以免费观看大片| 国模一区二区三区四区视频| kizo精华| 啦啦啦在线观看免费高清www| 免费看日本二区| 亚洲第一区二区三区不卡| 久久99热这里只频精品6学生| 久久精品国产a三级三级三级| 大话2 男鬼变身卡| 91精品伊人久久大香线蕉| 欧美一级a爱片免费观看看| 日本av免费视频播放| 日本欧美国产在线视频| 亚洲中文av在线| 成人18禁高潮啪啪吃奶动态图 | av网站免费在线观看视频| 久久青草综合色| 日韩视频在线欧美| 国产欧美日韩综合在线一区二区 | 18禁在线无遮挡免费观看视频| 欧美精品高潮呻吟av久久| 一区在线观看完整版| 99久久综合免费| 国产伦理片在线播放av一区| 国产欧美另类精品又又久久亚洲欧美| 最近的中文字幕免费完整| 黑丝袜美女国产一区| 青青草视频在线视频观看| 国产色婷婷99| 91aial.com中文字幕在线观看| 人妻人人澡人人爽人人| 蜜桃在线观看..| xxx大片免费视频| 久久99热6这里只有精品| 夫妻性生交免费视频一级片| 丰满乱子伦码专区| videos熟女内射| 一区在线观看完整版| 国产中年淑女户外野战色| 边亲边吃奶的免费视频| 春色校园在线视频观看| 夫妻性生交免费视频一级片| 亚洲色图综合在线观看| 国产精品99久久久久久久久| 搡老乐熟女国产| 国产午夜精品一二区理论片| 亚洲内射少妇av| 夜夜爽夜夜爽视频| 久久久久视频综合| av在线老鸭窝| 国产成人91sexporn| 亚洲av在线观看美女高潮| 国产伦精品一区二区三区视频9| 欧美丝袜亚洲另类| 国产 一区精品| 香蕉精品网在线| 婷婷色综合大香蕉| 妹子高潮喷水视频| 观看av在线不卡| 国产91av在线免费观看| 视频区图区小说| 日日啪夜夜爽| 丰满人妻一区二区三区视频av| 新久久久久国产一级毛片| 少妇人妻 视频| 三上悠亚av全集在线观看 | 插阴视频在线观看视频| 欧美区成人在线视频| 亚洲综合精品二区| 亚洲欧美精品自产自拍| 麻豆成人午夜福利视频| 交换朋友夫妻互换小说| 狂野欧美白嫩少妇大欣赏| 亚洲精品aⅴ在线观看| 熟女人妻精品中文字幕| 久久热精品热| av女优亚洲男人天堂| 日韩一本色道免费dvd| a级毛色黄片| 深夜a级毛片| 纯流量卡能插随身wifi吗| 国产精品秋霞免费鲁丝片| 日日啪夜夜爽| 亚洲自偷自拍三级| 久久久久久久久久久久大奶| 免费黄色在线免费观看| 日本与韩国留学比较| 91精品国产九色| 精品国产一区二区久久| 乱码一卡2卡4卡精品|