Network based subcellular proteomics in monocyte membrane revealed novel candidate genes involved in osteoporosis.

In this study, label-free-based quantitative subcellular proteomics integrated with network analysis highlighted several candidate genes including P4HB, ITGB1, CD36, and ACTN1 that may be involved in osteoporosis. All of them are predicted as significant membrane proteins with high confidence and enriched in bone-related biological process. The results were further verified in transcriptomic and genomic levels. INTRODUCTION: Osteoporosis is a metabolic bone disease mainly characterized by low bone mineral density (BMD). As the precursors of osteoclasts, peripheral blood monocytes (PBMs) are supported to be important candidates for identifying genes related to osteoporosis. We performed subcellular proteomics study to identify significant membrane proteins that involved in osteoporosis. METHODS: To investigate the association between monocytes, membrane proteins, and osteoporosis, we performed label-free quantitative subcellular proteomics in 59 male subjects with discordant BMD levels, with 30 high vs. 29 low BMD subjects. Subsequently, we performed integrated gene enrichment analysis, functional annotation, and pathway and network analysis based on multiple bioinformatics tools. RESULTS: A total of 1070 membrane proteins were identified and quantified. By comparing the proteins' expression level, we found 36 proteins that were differentially expressed between high and low BMD groups. Protein localization prediction supported the notion that the differentially expressed proteins, P4HB (p = 0.0021), CD36 (p = 0.0104), ACTN1 (p = 0.0381), and ITGB1 (p = 0.0385), are significant membrane proteins. Functional annotation and pathway and network analysis highlighted that P4HB, ITGB1, CD36, and ACTN1 are enriched in osteoporosis-related pathways and terms including "ECM-receptor interaction," "calcium ion binding," "leukocyte transendothelial migration," and "reduction of cytosolic calcium levels." Results from transcriptomic and genomic levels provided additional supporting evidences. CONCLUSION: Our study strongly supports the significance of the genes P4HB, ITGB1, CD36, and ACTN1 to the etiology of osteoporosis risk.

[Cone beam CT-derived adaptive radiotherapy for setup error assessment and correction in whole breast intensity modulated radiotherapy].

OBJECTIVE: To quantify the setup error (SE) in breast cancer patients treated with intensity modulated radiotherapy (IMRT) based on cone beam CT (CBCT), and to explore the feasibility of using several CBCT scans to presume and correct SE in the treatment for breast cancer patients. METHODS: Eighteen breast cancer patients after breast conserving surgery who underwent whole breast IMRT were included in this study. Three dimensional interfraction motion before and after on-line CBCT-based corrections were quantified. The on-line CBCT-based corrections were performed using automated greyscale match. The system SE (Σ) and random error (σ) were calculated for each patient based on the consecutive multiple online scanning based on CBCT (≥5). The trends in magnitudes of Σ and σwere assessed during the treatment. RESULTS: The magnitude variation of Σ was less than 1 mm before and after on-line CBCT-based corrections. As the CBCT scanning times increase (before 10 times), the Σ in anteroposterior (AP) direction was increased significantly, and σin three dimensional directions was also increased after 7 times of CBCT scanning. After on-line CBCT-based corrections, the Σ showed a steady trend by variation near zero for the first 20 times irradiation; but after 20 times, the Σ in AP and superoinferior (SI) directions was increased slightly (less than 0.5 mm), and σdecreased in three-dimensional directions. There were no significant differences for Σ, σand setup margin (SM) before and after on-line CBCT-based corrections in all three directions (P>0.05). CONCLUSIONS: For breast cancer patients who underwent IMRT after breast conserving surgery, the setup error is relatively stable during the whole irradiation. The first 5 CBCT scans are suitable to presume and correct SE, and also can be used as the right time for adaptive radiotherapy planning revision.

A follow-up association study of two genetic variants for bone mineral density variation in Caucasians.

SUMMARY: We tested whether two genetic variants were associated with BMD at multiple clinically relevant skeletal sites in Caucasians. We found that variant rs7776725 is consistently associated with hip, spine, wrist and whole-body BMD, which highlights the potential importance of this variant or linked variants for osteoporosis. INTRODUCTION: A recent genome-wide association study identified two single nucleotide polymorphisms (SNPs), rs7776725 and rs1721400, that were associated with bone mineral density (BMD) variation at the radius, tibia and calcaneus in a Korean population. In this study, we aimed to test whether the association of these two genetic variants can be replicated in Caucasians and whether their association with BMD can be extended to other clinically relevant skeletal sites. METHODS: We performed this study in two large cohorts of unrelated US Caucasians. Area BMD at the hip, spine, wrist (ultra-distal radius) and whole body were measured with Hologic dual-energy X-ray absorptiometer. SNPs were genotyped with Affymetrix human genome-wide genotyping arrays. Association analyses were performed using PLINK. RESULTS: We detected highly significant association (combined p = 1.42 × 10(-16)) of rs7776725 with wrist BMD but only borderline association signal (combined p = 0.017) for rs1721400 with wrist BMD. In addition, we found that rs7776725 was associated with BMD at the hip, spine and whole body. At the FAM3C gene locus where rs7776725 was located, we identified several other SNPs (rs4727922, rs1803389, rs718766 and rs7793554) that were also associated with BMD. CONCLUSIONS: This is the first follow-up association study of rs7776725 and rs1721400 with BMD. The rs7776725 showed consistent association with BMD at multiple clinically important skeletal sites, which highlighted the potential importance of rs7776725 or linked SNPs for risk of osteoporosis. Further in-depth re-sequencing studies and functional assays are necessary to elucidate the underlying mechanisms.

Pharmacokinetics and tissue distribution of zidovudine in rats following intravenous administration of zidovudine myristate loaded liposomes.

Liposomes accumulating in the reticuloendothelial system (RES) appear to be a promising vehicle to improve the therapeutic index of anti-HIV drugs such as zidovudine (AZT). Since the entrapment efficiency of AZT in liposomes was found to be low and AZT leakage from liposomes is fast, zidovudine myristate (AZT-M) was synthesized as a prodrug, and AZT-M incorporated liposomes in a lyophilized form were prepared with an average diameter of 90 nm and an encapsulation efficiency of 98% after reconstitution. The pharmacokinetic profiles and tissue distribution of AZT after i.v. administration of AZT-M liposomes in rats were investigated, and the results were compared with those after i.v. administration of AZT solution. AZT levels in plasma were significantly higher following application of AZT-M liposomes compared with AZT solution, and AUC0_infinity increased from 5.0 +/- 0.7 micromol x min x ml(-1) to 8.2 +/- 1.7 micromol x min x ml(-1) accordingly. Tissue distribution studies also confirmed higher concentrations of AZT in organs of RES and brain, suggesting that AZT-M liposomes might be promising candidates for therapy of HIV infections.

Serum and tissue expression of gelatinase and Twist in breast cancer.

OBJECTIVE: This study aimed to investigate the levels of matrix metalloproteinases (MMP)-2, MMP-9 and Twist in tumor tissue and serum from 46 cases of breast cancer patients and 31 cases of benign breast diseases patients by immunohistochemical staining and enzyme-linked immunosorbent assay, respectively. The association of gelatinase and Twist expression with clinicopathological factors was also analyzed in the present study. PATIENTS AND METHODS: The studied population consisted of 46 breast cancer patients and 31 benign breast disease patients. Serum concentrations of MMP-2, MMP-9 and Twist were measured by using human enzyme-linked immunosorbent assay. The protein expression of Twist, MMP-2 and MMP-9 were determined by immunohistochemical. RESULTS: The results show that the levels of MMP-2, MMP-9 and Twist expression were significantly increased in tissue and serum from breast cancer group, compared to the group of benign breast lesions diseases (p < 0.05). The pre-operative serum levels of MMP-2, MMP-9 and Twist were positively correlated with their expression in breast cancer tissues, respectively (p < 0.05). We, then, correlated serum and tissue levels of MMP-2, MMP-9 and Twist in breast cancer samples with patients' clinicopathologic characteristics. Compared to low expression, high serum and tissue levels of MMP-2 and Twist were associated with lymph node metastasis and higher TNM stage, high tissue MMP-9 levels were associated with lymph node metastasis and higher TNM stage, and high serum MMP-9 levels were associated with c-erbB-2 expression. CONCLUSIONS: These data suggest that serum levels of MMP-2, MMP-9 and Twist could be as potential biomarkers for diagnosis and predicting metastasis of breast cancer.

Phylogenetic relationships among two species of golden monkey and three species of leaf monkey inferred from rDNA variation.

Restriction maps of rDNA repeats of five species of Colobinae and three outgroup taxa, Hylobates leucogenys, Macaca mulatta, and Macaca irus, were constructed using 15 restriction endonucleases and cloned 18S and 28S rRNA gene probes. The site variation between Rhinopithecus roxellana and Rhinopithecus bieti is comparable to that between Presbytis françoisi and Preshytis phayrei, implying that R. bieti is a valid species rather than a subspecies of R. roxellana. Phylogenetic analysis on the 47 informative sites supports the case for Rhinopithecus being an independent genus and closely related to Presbytis. Furthermore, branch lengths of the tree seem to support the hypothesis that the leaf monkeys share some ancestral traits as well as some automorphic characters.