冠心病患者外周血单个核细胞YKT6基因表达及其临床意义

目的：研究YKT6在冠心病（coronary heart disease,CHD）患者外周血单个核细胞中的表达及其与疾病发生、发展的关系.方法：分别采用半定量逆转录-聚合酶链反应（RT-PCR）和Westen blot检测40例CHD患者和37例健康者YKT6 mRNA和蛋白表达.结果：CHD患者外周血单个核细胞中YKT6 mRNA和蛋白表达均明显高于健康者（P＜0.05）.结论：YKT6表达与CHD发生存在一定相关性,在CHD发病过程中可能起重要作用,为CHD的诊断和预防提供了新线索.     

Aberrant trafficking of the high-affinity choline transporter in AP-3-deficient mice

The high-affinity choline transporter (CHT) is expressed in cholinergic neurons and efficiently transported to axon terminals where it controls the rate-limiting step in acetylcholine synthesis. Recent studies have shown that the majority of CHT is unexpectedly localized on synaptic vesicles (SV) rather than the presynaptic plasma membrane, establishing vesicular CHT trafficking as a basis for activity-dependent CHT regulation. Here, we analyse the intracellular distribution of CHT in the adaptor protein-3 (AP-3)-deficient mouse model mocha . In the mocha mouse, granular structures in cell bodies are intensely labelled with CHT antibody, indicating possible deficits in CHT trafficking from the cell body to the axon terminal. Western blot analyses reveal that CHT on SV in mocha mice is decreased by 30% compared with wild-type mice. However, no significant difference in synaptosomal choline uptake activity is detected, consistent with the existence of a large reservoir pool for CHT. To further characterize CHT trafficking, we established a PC12D-CHT cell line. In this line, CHT is found associated with a subpopulation of synaptophysin-positive synaptic-like microvesicles (SLMV). The amounts of CHT detected on SLMV are greatly reduced by treating the cell with agents that halt AP-dependent membrane trafficking. These results demonstrate that APs have important functions for CHT trafficking in neuronal cells.     

An alternative mechanism of early nodal clustering and myelination onset in GABAergic neurons of the central nervous system

In vertebrates, fast saltatory conduction along myelinated axons relies on the node of Ranvier. How nodes assemble on CNS neurons is not yet fully understood. We previously described that node-like clusters can form prior to myelin deposition in hippocampal GABAergic neurons and are associated with increased conduction velocity. Here, we used a live imaging approach to characterize the intrinsic mechanisms underlying the assembly of these clusters prior to myelination. We first demonstrated that their components can partially preassemble prior to membrane targeting and determined the molecular motors involved in their trafficking. We then demonstrated the key role of the protein β2Na_v for node-like clustering initiation. We further assessed the fate of these clusters when myelination proceeds. Our results shed light on the intrinsic mechanisms involved in node-like clustering prior to myelination and unravel a potential role of these clusters in node of Ranvier formation and in guiding myelination onset.     

PLAC1 mRNA levels in maternal blood at induction of labor correlate negatively with induction-delivery interval

OBJECTIVE: This study was conducted to determine whether, in low risk women having labor induced using prostaglandin gel (dinoprostone gel), there is a relationship between the concentration of mRNA for the PLAC1 gene (a trophoblast-specific gene) in maternal blood and the time elapsed between the first gel administration and spontaneous delivery. STUDY DESIGN: Blood was collected from 49 selected women at 40.2-41.4 weeks' gestation. Total RNA was extracted by means of an ABI Prism 6100 nucleic acid Prep Station and quantitative real-time PCR analysis was performed by use of a PE Applied Biosystems 5700 Sequence Detection System. Sequence data were obtained from the Genebank Sequence Database. To determine the amount of cDNA, the PLAC1 locus was used. RESULTS: Thirty women (61.2%) had a spontaneous delivery. A caesarean section, either for fetal dystocia or fetal distress, was performed in 19 (38.8%). The crude delivery rates of the women who ended up with a spontaneous delivery were 30% at 24 h and 43% at 48 h. Women (n=19) with a blood concentration of logPLAC1 mRNA>or=2.00 displayed a median time to delivery of 23.50h, (95% CI: 13.13-33.87) while those with a logPLAC1 mRNA<2.00 (n=30) had a median time of 54 h. (95% CI: 37.86-70.14; p=0.0043, log-rank test). By means of multivariate analysis, quantitative Bishop score (from 2 to 7) at the time of the first gel administration and logPLAC1 mRNA>or=2.00 were associated with a higher rate of delivery per unit of time with an odds ratio of 1.35 (95% CI: 1.07-1.71) and 3.48 (95% CI: 1.55-7.80), respectively. CONCLUSIONS: In induced term pregnancies, PLAC1 mRNA in maternal blood at the beginning of the treatment correlates with the time elapsed before delivery. This evidence demonstrates that the fetomaternal trafficking of nucleic acids is more consistent when the labor is about to begin.     

Mental health of female survivors of human trafficking in Nepal

Little is known about the mental health status of trafficked women, even though international conventions require that it be considered. This study, therefore, aims at exploring the mental health status, including anxiety, depression and post-traumatic stress disorder (PTSD), of female survivors of human trafficking who are currently supported by local non-governmental organizations (NGOs) in Katmandu, the capital of Nepal, through comparison between those who were forced to work as sex workers and those who worked in other areas such as domestic and circus work (non-sex workers group). The Hopkins Symptoms Checklist-25 (HSCL-25) was administered to assess anxiety and depression, and the PTSD Checklist Civilian Version (PCL-C) was used to evaluate PTSD. Both the sex workers' and the non-sex workers' groups had a high proportion of cases with anxiety, depression, and PTSD. The sex workers group tended to have more anxiety symptoms (97.7%) than the non-sex workers group (87.5%). Regarding depression, all the constituents of the sex workers group scored over the cut-off point (100%), and the group showed a significantly higher prevalence than the non-sex workers (80.8%). The proportion of those who are above the cut-off for PTSD was higher in the sex workers group (29.6%) than in the non-sex workers group (7.5%). There was a higher rate of HIV infection in the sex workers group (29.6%) than in the non-sex workers group (0%). The findings suggest that programs to address human trafficking should include interventions (such as psychosocial support) to improve survivors' mental health status, paying attention to the category of work performed during the trafficking period. In particular, the current efforts of the United Nations and various NGOs that help survivors of human trafficking need to more explicitly focus on mental health and psychosocial support.     

Glycosylation Regulates Prestin Cellular Activity

Glycosylation is a common post-translational modification of proteins and is implicated in a variety of cellular functions including protein folding, degradation, sorting and trafficking, and membrane protein recycling. The membrane protein prestin is an essential component of the membrane-based motor driving electromotility changes (electromotility) in the outer hair cell (OHC), a central process in auditory transduction. Prestin was earlier identified to possess two N-glycosylation sites (N163, N166) that, when mutated, marginally affect prestin nonlinear capacitance (NLC) function in cultured cells. Here, we show that the double mutant prestin^NN163/166AA is not glycosylated and shows the expected NLC properties in the untreated and cholesterol-depleted HEK 293 cell model. In addition, unlike WT prestin that readily forms oligomers, prestin^NN163/166AA is enriched as monomers and more mobile in the plasma membrane, suggesting that oligomerization of prestin is dependent on glycosylation but is not essential for the generation of NLC in HEK 293 cells. However, in the presence of increased membrane cholesterol, unlike the hyperpolarizing shift in NLC seen with WT prestin, cells expressing prestin^NN163/166AA exhibit a linear capacitance function. In an attempt to explain this finding, we discovered that both WT prestin and prestin^NN163/166AA participate in cholesterol-dependent cellular trafficking. In contrast to WT prestin, prestin^NN163/166AA shows a significant cholesterol-dependent decrease in cell-surface expression, which may explain the loss of NLC function. Based on our observations, we conclude that glycosylation regulates self-association and cellular trafficking of prestin^NN163/166AA. These observations are the first to implicate a regulatory role for cellular trafficking and sorting in prestin function. We speculate that the cholesterol regulation of prestin occurs through localization to and internalization from membrane microdomains by clathrin- and caveolin-dependent mechanisms.     

Sex work in Tallinn, Estonia: the sociospatial penetration of sex work into society

BACKGROUND: It is important to describe and understand the underlying patterns and dynamics that govern sex work in societies undergoing rapid political and social changes, its heterogeneity across populations, and its evolution through time in order to inform future research, sound policy formation, and programme delivery. OBJECTIVES: To describe the socioeconomic and cultural determinants, organisational structure, distinct categories, and spatial patterning of sex work in Tallinn, Estonia, and identify recent temporal changes in sex work patterns. METHODS: In-depth interviews with key informants; naturalistic observations of sex work and drug use venues, geo-mapping of sex work sites, review of media, public policy, and commissioned reports, and analyses of existing data. RESULTS: Sex work takes place in a hierarchy of locations in Tallinn ranging from elite brothels and "love flats" to truck stops. These sites vary in terms of their public health importance and social organisation. There are full time, part time, and intermittent male and female sex workers. Among others, the taxi driver, madam and the bartender are central roles in the organisation of sex work in Tallinn. Cell phone and internet technology enable sex work to be highly dispersed and spatially mobile. CONCLUSION: Future research and programmatic service delivery or outreach efforts should respond to the changing profile of sex work in Tallinn and its implications for STD/HIV epidemiology.     

Ex vivo expansion of hematopoietic progenitor cells is associated with downregulation of α4 integrin- and CXCR4-mediated engraftment in NOD/SCID β2-microglobulin-null mice

Background

Several studies indicate that ex vivo cytokine-supported expansion induces defective hematopoietic stem cell engraftment. We investigated the role of α4 integrin, α5 integrin and CXCR4 in engraftment of unmanipulated and cytokine-treated human cord blood CD34⁺ cells.

Design and Methods

Uncultured or expanded CD34⁺ cells were infused in NOD/SCID-β₂microglobulin-null mice. The function of α4, and α5 integrins and CXCR4 was assessed by incubating cells with specific neutralizing antibodies, prior to transplant. The activation state of α4 integrin was further tested by adhesion and migration assays.

Results

Neutralization of either α4 integrin or CXCR4 abolished engraftment of uncultured CD34⁺ cells at 6 week spost-transplant, while α5 integrin neutralization had no significant effect. However, after short-term ex vivo culture, blocking α4 integrin or CXCR4 did not affect repopulating activity whereas neutralization of α5 integrin inhibited engraftment. Using soluble vascular cell adhesion molecule-1 binding assays, we observed that α4 integrin affinity in fresh CD34⁺ cells was low and susceptible to stimulation while in cultured CD34⁺ cells, it was high and insensitive to further activation. In addition, stromal cell-derived factor-1 stimulated migration across vascular cell adhesion molecule-1 in fresh CD34⁺ cells but not in cultured CD34⁺ cells.

Conclusions

Our data show that ex vivo culture of hematopoietic progenitor cells is associated with downregulation of both α4 integrin- and CXCR4-mediated engraftment. Further investigations suggest that this is caused by supraphysiological increase of α4 integrin affinity, which impairs directional migration across vascular cell adhesion molecule-1 in response to stromal cell-derived factor-1. Such changes may underlie the engraftment defect of cytokine-stimulated CD34⁺ cells.     

Recombinant IL‐7/HGFβ hybrid cytokine separates acute graft‐versus‐host‐disease from graft‐versus‐tumour activity by altering donor T cell trafficking

Given that donor T cells from a transplant contribute both the desired graft‐versus‐tumour (GVT) effect and detrimental graft‐versus‐host disease (GVHD), strategies to separate GVHD and GVT activity are a major clinical goal. We have previously demonstrated that in vivo administration of a recombinant (r)IL‐7/HGFβ hybrid cytokine, consisting of interleukin‐7 (IL‐7, IL7) and the β‐chain of hepatocyte growth factor (HGFβ), significantly inhibits the growth of cancer cells in murine tumour models. The antit‐umour effect of rIL‐7/HGFβ is related to a marked infiltration T cells in the tumour tissues. We have also shown that GVHD was not induced in rIL‐7/HGFβ‐treated T cell‐depleted allogeneic haematopoietic stem cell transplantation (HSCT) recipients. We show here that, in T cell‐replete allogeneic HSCT murine models, rIL‐7/HGFβ attenuated acute GVHD (aGVHD), while promoting GVT activity. This was related to an alteration of donor T cell trafficking, with an increased infiltration of donor T cells into tumour tissues and the lympho‐haematopoietic system but decreased number of the T cells in the GVHD target organs. Therefore, rIL‐7/HGFβ may offer a new tool to alleviate aGVHD while prompting GVT, and to study the molecular regulation of T cell trafficking.