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Objectives: To determine the association between obesity and bone mineral density in menopausal women., Methods: We recruited menopausal women aged 50 years and older who undertook health examinations in West China Hospital of Sichuan University in this study. Bone mineral density of the participants was measured by MetriScan Bone Densitometry. The participants were categorized into three groups according to the T value: normal density, osteopenia and osteoporosis., Results: Of the 4 938 participants, 8.55% had obesity [body mass index (BMI)>28 kg/m 2 ]. The three groups of participants were different in BMI, a body shape index (ABSI), body mass, waist circumference, hip circumference and height ( P <0.01). The age-adjusted T values were positively correlated with height, body mass, waist circumference, hip circumference, waist-hip ratio (WHR), waist-height ratio (WHtR) and body roundness index (BRI) ( P <0.05). The areas under curves ( AUC ) of receiver operator characteristic (ROC) were 0.540 and 0.568, respectively, for waist circumference and BMI in those with osteoporosis., Conclusions: Obesity in menopausal women is negatively associated with osteoporosis. The clinical significance of such an association requires further studies with a longitudinal design.

Unlabelled: OBJECTIVE To construct the recombinant plasmid pCI-HLE encoding human serum album-EPO (HSA-EPO) fusion protein and to express it in CHO cell., Methods: The cDNA encoding human serum album and EPO were amplified by PCR, and then spliced with the synsitic DNA fragment encoding GS (GGGGS), by overlap PCR extension to form LEPO. After BamH I digestion, the HSA and LEPO was ligated to generate the fusion HSA-EPO gene and was then cloned into the expression vector pCI-neo to generate the recombinant plasmid pCI-HLE. The plasmid pCI-HLE was transfected into CHO cell by liposome protocol. Then, the recombinant cells were screened by G418 and identified by PCR and Western blot. Expression of fusion protein was evaluated by Enzyme Linked Immunosorbent Assay (ELISA)., Results: Restrictive enzymes digestion and DNA sequencing revealed that HSA-EPO fusion gene was cloned into expression vector pCI-neo successfully. PCR and Western blot analysis confirmed that the fusion gene was integrated in the genome of CHO cells and expressed successfully. The HSA-EPO production varied from 86 Iu/(mL x 10(6) x 72 h) to 637 IU/(mLx 10(6) x 72 h)., Conclusion: The results confirmed that HSA-EPO fusion gene can be expressed in the CHO cells, with EPO immunogenicity, which could serve as foundation for the development of long-lasting recombinant HSA-EPO protein.

Objective: To study the influence and mechanism of hypoxia on the expression of angiotensin-converting enzyme 2 (ACE2) in human umbilical vein endothelial cells (HUVECs)., Methods: HUVECs were cultured as adherent cells and identified by immunohistochemical study of Factor VIII. Then the cells were cultured in 37 degrees C incubators under hypoxia condition (1% O2 + 94% N2 + 5%C O2) or regular 5% CO2 condition for 3 h, 6 h, 12 h, 24 h, respectively, and the culture medium and cells were collected at each time point. ACE2 protein was measure by immunohistochemical assay, ACE2 mRNA was quantified by real-time RT-PCR., Results: (1) ACE2 protein expression decreased after hypoxia treatment for 3 h, 6 h, 12 h, and 24 hour compared with regular oxygen condition (chi2 = 825.078, 768.141, 623.931, 350.260, P < 0.00625), the maximum reduction came at 3 h after hypoxia exposure, then raised gradually after 6 h, 12 h, and 24 h exposure; (2) ACE2 mRNA expression show a similar tendency to ACE2 protein. After hypoxia disposal, its relative expression rate reduced to 0.281 at 3 h (P = 0.031), then increased gradually at 6 h (0.445, P = 0.034), 12 h (0.794, P = 0.434), 24 h (0.891, P = 0.692)., Conclusion: Our study show hypoxia down-regulate the protein and gene expression of ACE2 in HUVECs in 24 hours. The down-regulation effect of hypoxia reached maximum level at 3 h, and then attenuated gradually at 6 h, 12 h and 24 h, which indicate that ACE2 might play an important role in the regulation of RAS in HUVECs after hypoxia injury.

Objective: To investigate the expression of AQP1 in endothelial cells under hypoxia conditions in vitro., Methods: The cultured human umbilical vein endothelial cells were randomly assigned to hypoxia groups (exposed to 1% of O2 for 3 h, 6 h, 12 h and 24 h respectively), and control group (ordinary culture). The expressions of AQP1 mRNA and protein were detected by real time PCR and immunohistochemistry respectively., Results: The expressions of AQP1 protein and mRNA increased after exposure to hypoxia initially and then decreased afterwards., Conclusion: Hypoxia regulates the expression of AQP1 in vascular endothelial cells.