Molecular characterization of a MOSPD2 homolog in the barbel steed (Hemibarbus labeo) and its involvement in monocyte/macrophage and neutrophil migration.

Motile sperm domain containing 2 (MOSPD2) is a single-pass membrane protein to which until recently little function had been ascribed. Although its mammalian homologs have been identified, the status of the mospd2 gene in lower vertebrates is still unknown. In the present study, cDNA of the mospd2 gene of barbel steed (Hemibarbus labeo) was cloned and sequenced to characterize its potential involvement in the innate immune system of this fish. Sequence analysis revealed that the predicted barbel steed MOSPD2 protein contained an N-terminal extracellular portion composed of a CRAL-TRIO domain, a motile sperm domain, and a transmembrane domain, as well as a short C-terminal intracellular domain. Phylogenetic tree analysis indicated that barbel steed MOSPD2 is closely related to that of zebrafish. Barbel steed mospd2 transcripts were detected in a wide range of tissues, with the highest level being found in the gill. In response to lipopolysaccharide (LPS) treatment or Aeromonas hydrophila infection, mospd2 gene expression was significantly altered in the head kidney, spleen, and mid-intestine. The expression of mospd2 gene was detected in monocytes/macrophages (MO/MФ), neutrophils, and lymphocytes, and was found to be mainly expressed in MO/MФ. At the same time, using flow cytometry, we also confirmed that MOSPD2 protein is located on MO/MФ, neutrophil, and lymphocyte membranes. Following treatment with LPS or A. hydrophila, MOSPD2 protein expression was induced in these immune cells. The migration of MO/MФ and neutrophils decreased significantly upon MOSPD2 blockade with anti-MOSPD2 IgG in a dose-dependent manner, whereas this treatment had no significant effect on lymphocytes migration. To the best of our knowledge, our study, for the first time, provides evidence that MOSPD2 mediates the migration of MO/MФ and neutrophils in a fish species.

Intratumoral estrogen sulfotransferase induction contributes to the anti-breast cancer effects of the dithiocarbamate derivative TM208.

AIM: Sulfotransferase-catalyzed sulfation is the most important pathway for inactivating estrogens. Thus, activation of estrogen sulfotransferase (EST) may be an alternative approach for the treatment of estrogen-dependent breast cancer. In this study we investigated the involvement of EST in anti-breast cancer effects of the dithiocarbamate derivative TM208 in vitro and in vivo. METHODS: The viability of human breast cancer MCF-7 cells was determined using a SBB assay. Nude mice bearing MCF-7 cells were orally administered TM208 (50 and 150 mg·kg(-1)·d(-1)) for 18 days. The xenograft tumors and uteri were collected. The mRNA expression of EST was examined with real-time PCR. EST protein was detected with Western blot, ELISA or immunohistochemical staining assays. A radioactive assay was used to measure the EST activity. Uterotropic bioassay was used to examine the uterine estrogen responses. RESULTS: Treatment with TM208 (10, 15 and 20 µmol/L) concentration-dependently increased EST expression in MCF-7 cells in vitro. Co-treatment with triclosan, an inhibitor of sulfonation, abolished TM208-induced cytotoxicity in MCF-7 cells. TM208 exhibited an apparent anti-estrogenic property: it exerted more potent cytotoxicity in E2-treated MCF-7 cells. In the nude mice bearing MCF-7 cells, TM208 administration time-dependently increased the expression and activity of EST, and blocked the gradual increase of E2 concentration in the xenograft tumors. Furthermore, TM208 administration blocked the estrogens-stimulated uterine enlargement. Tamoxifen, a positive control drug, produced similar effects on the expression and activity of EST in vitro and in vivo. CONCLUSION: The induction of EST and reduction of estrogen concentration contribute to the anti-breast cancer action of TM208 and tamoxifen. TM208 may be developed as anticancer drug for the treatment of estrogen receptor-positive breast cancer.

Dopamine D1 receptor activation induces dehydroepiandrosterone sulfotransferase (SULT2A1) in HepG2 cells.

AIM: Dopamine receptors are present in the nervous system and also widely distributed in the periphery. The aim of this study was to investigate the role of D1 subtype dopamine receptors (DRD1) in the regulation of dehydroepiandrosterone sulfotransferase (SULT2A1) in HepG2 cells. METHODS: HepG2 cells were treated with DRD1 agonists with or without DRD1 antagonist for 9 d. DRD1 and SULT2A1 mRNA expression, protein expression, and SULT2A1 activity were detected using RT-PCR, Western blotting and HPLC, respectively. The level of cAMP was measured using a commercial kit. RESULTS: All the 5 DR subtypes (DRD1-DRD5) were found to be expressed in HepG2 cells. Treatment of HepG2 cells with the specific DRD1 agonists SKF82958 (2.5 µmol/L) or SKF38393 (5 and 50 µmol/L) significantly increased the mRNA and protein expression of both DRD1 and SULT2A1, and increased SULT2A1 activity and cAMP levels. These effects were partially blocked by co-treatment with the specific DRD1 antagonist SCH23390 (2.5 µmol/L). In addition, transfection of HepG2 cells with DRD1-specific siRNAs decreased DRD1 mRNA expression by 40%, which resulted in the reduction of SULT2A1 mRNA expression by 60%, protein expression by 40%, and enzyme activity by 20%. CONCLUSION: DRD1 activation upregulates DRD1 and SULT2A1 expression and SULT2A1 activity in HepG2 cells, suggesting that the DRD1 subtype may be involved in the metabolism of drugs and xenobiotics through regulating SULT2A1.

[Soil respiration characteristics in the clear-cutting site of Quercus aliena var. acuteserrata forest in Xiaolong Mountain in Qinling Mountains].

By using Li-6400 portable photosynthesis system with soil chamber, the soil respiration rates (Rs) of Quercus aliena var. acuteserrata forest land (control site) and their clear-cutting site were measured in Xiaolong Mountain of Qinling Mountains from May 2011 to April 2012 to understand the diurnal and monthly dynamics of soil respiration rate and the influences from soil temperature, soil moisture, soil physical and chemical properties. The results showed that both diurnal and monthly dynamics of soil respiration rate presented a single-peak curve, similar to the variation of soil temperature at the clear-cutting and control sites. During the study period, the maximum monthly mean values of soil respiration rate at the clear-cutting and control sites occurred in July (4.63 and 4.01 micromol x m(-2) x s(-1), respectively) and the minimum values presented in February (0.10 and 0.30 micromol x m(-2) x s(-1), respectively). Soil respiration rate in 4-6 months after clear-cutting was higher than at the control site, and became lower afterwards. 89. 6% -90. 8% of soil respiration rate variation was interpreted by the multiple regression models of soil temperature, soil moisture and their interaction at the clear-cutting site, and 94.7%-95.5% at the control site. The Q10 values computed by exponential equations were 3.47-4.22 and 3.54-3.96 at the clear-cutting and control sites, respectively. The C fluxes at the clear-cutting and control sites were 344.8 and 512.9 g x m(-2) annually, and 24.2 and 40.9 g x m(-2) in winter, respectively.

[Combined treatment with areola approach for capsular contracture after breast augmentation with implants].

OBJECTIVE: To investigate the combined treatment with areola approach for capsular contracture after breast augmentation with implants. METHODS: From Feb. 2005 to Jun. 2011, 94 cases (168 sides) with Baker III and IV capsular contracture after breast augmentation with implants were treated with areola approach. The implants cavity was recreated, with or without removal of capsule. The implants were reimplanted behind pectoralis major or breast at the second stage in some patients. RESULTS: 46 cases were followed up by clinic visit and the others were followed up by telephone for 6-37 months, with an average of 9.9 months. The capsular contracture was relapsed in 2 cases as Baker III and 1 case as Baker IV. All the other breasts got a good appearance with good soft texture and feeling. No hematoma, infection, implants rupture, breast ptosis or implant displacement happened. CONCLUSIONS: Combined treatment with areola approach has a good therapeutic effect for capsular contracture after breast augmentation with implants. The breast appearance is satisfactory with low occurrence of capsular contracture.

[Spiral computed tomography imaging of internal carotid artery diameter: a report of 260 cases].

OBJECTIVE: To explore the diameter of internal carotid artery by arterial phase imaging of multi-slice spiral computed tomography (CT). METHODS: A total of 260 routine brain CT scans of the examiner were performed. And the arterial phase images of carotid artery were acquired, observed and measured through a three-dimensional reconstruction workstation. On the diameters of target sections were measured on the multiplanar reconstruction (MPR) images of carotid artery. Two groups were categorized according to gender and 3 groups by age (25 - 40 yr, 41 - 60 yr and 61 - 85 yr) for statistical analysis. RESULTS: The diameter data of internal carotid artery had statistical significances among genders and 3 age groups (P < 0.05). CONCLUSION: The diameter of internal carotid artery may be evaluated by the arterial phase imaging of multi-slice spiral CT so that the reference data can be provided for clinical diagnosis.

[The telomerase activity of human adipose derived stem cells during proliferation and differentiation in vitro].

OBJECTIVE: To investigate the telomerase activity of human adipose derived stem cells (ADSCs) during proliferation and differentiation in vitro. METHODS: ADSCs were highly purified and cultured in vitro. The morphology, phenotype and biological properties of the cultured ADSCs were observed by flow cytometer. Then ADSCs were induced to differentiate into adipocytes and osteoblast. The telomerase activity was detected by TRAP. RESULTS: ADSCs had the ability of multi-directed differentiation, like adipocytes and osteoblast. It could also express the stem cell-related surface markers. The telomerase activity was negative or lowly expressed in ADSCs in vitro within 12 generations. The telomerase activity was up-regulated when ADSCs was adipogenic differentiated, but deceased 3-6 days later. CONCLUSIONS: The telomerase activity of ADSCs is not changed during culture in vitro. It is up-regulated when ADSCs are induced to adipogenic differentiation, but decreased later.

Significance of MMP-2 and TIMP-2 mRNA expressions on glomerular cells in the development of glomerulosclerosis.

OBJECTIVE: To study the expressions of MMP-2 and TIMP-2 mRNA on cultured rat mesangial cells (MsC) and in human diseased glomeruli, and to explore their significance in the development of glomerulosclerosis. METHODS: The expressions of MMP-2, TIMP-2, and Col IV mRNA on cultured rat MsC stimulated by IL-1 or/and TGF-beta1 were investigated through Northern blot analysis. The levels of MMP-2 and TIMP-2 mRNA expressions and immunoreactivity of PCNA and Col IV in human diseased glomeruli from renal biopsies of lupus nephritis (LN) patients were examined by in situ hybridization and immunohistochemistry, respectively. RESULTS: The levels of MMP-2, TIMP-2, and Col IV mRNA expressions were markedly increased on cultured rat MsC stimulated by IL-1 or/and TGF-beta1. Meanwhile, upregulation of MMP-2 and TIMP-2 mRNA expressions was confirmed in diseased glomeruli from patients with various subtypes of LN, and was closely related to the positive cell number of PCNA presentation and deposition of Col IV in glomeruli. CONCLUSION: The results suggest that the over-expressions of MMP-2 and TIMP-2 mRNA on glomerular cells might play a critical role in the development of glomerulosclerosis.