OBJECTIVES:The combined effects of physical activity and sleep duration on depressive symptoms trajectories, and age and gender disparities remain inadequately understood. The objective of this study was to identify the joint relationship of physical activity and sleep duration with depressive symptoms trajectories in Chinese middle-aged and older adults, and explored disparities in the joint associations. METHOD:This longitudinal analysis draws upon data from the China Health and Retirement Longitudinal Study. The analysis comprised 35063 observations. Linear mixed-effect models were employed to discern independent and joint correlation of physical activity and sleep duration with depressive symptoms trajectories. RESULTS:Chinese middle-aged and older adults showed increasing depressive symptoms trajectory. Physical activity and sleep duration interplayed to impact depressive symptoms trajectory in different ways. Compared to participants engaged in lower physical activity and short sleep, depressive symptom trajectories of those with higher physical activity and optimal sleep, and higher physical activity and short sleep increased less rapidly among group aged 60-70 years, as well as in women group. CONCLUSION:This study emphasized the importance of combining interventions targeting habits of physical activity and sleep for middle-aged and older adults to improve their mental health, especially for women and those aged 60-70 years.

Dysregulated IL-10 producing regulatory B cells (Bregs) are associated with the progression of systemic lupus erythematosus. An immunomodulatory role of heat shock proteins (HSPs) is implicated in autoimmune diseases. However, the molecular basis underlying the role of Hspa13 in regulating Bregs function and lupus pathogenesis remains unclear. In this study, Bregs display higher Hspa13 expression than IL-10- B cells. Induction of IL-10 production is weakened in B cells with Hspa13 knockdown or knockout. Hspa13 binds to the IL-10 promoter via the TATA or CAAT box and activates IL-10 transcription in the nucleus. Furthermore, Hspa13 positive cells are enriched in marginal zone (MZ) B cells to regulate IL-10 production. Stimulated B220+ B or MZ B cells from CD19creHspa13fl/fl mice for Breg induction show an impaired capacity to promote CD4+Foxp3+ regulatory T cells (Treg) differentiation. In lupus MRL/lpr mice, a decline in Treg differentiation is accompanied by decreased Hspa13 expression in both Bregs and MZ B cells. Moreover, adoptive transfusion of Bregs and MZ B cells from CD19creHspa13fl/fl mice fails to increase the frequency of Tregs, attenuate renal pathology, or decrease anti-dsDNA antibody levels. These results explain the unique role of Hspa13 in determining MZ regulatory function and affecting lupus pathogenesis.

Bile acids (BA) are important physiological molecules not only mediating nutrients absorption and metabolism in peripheral tissues, but exerting neuromodulation effect in the central nerve system (CNS). The catabolism of cholesterol to BA occurs predominantly in the liver by the classical and alternative pathways, or in the brain initiated by the neuronal-specific enzyme CYP46A1 mediated pathway. Circulating BA could cross the blood brain barrier (BBB) and reach the CNS through passive diffusion or BA transporters. Brain BA might trigger direct signal through activating membrane and nucleus receptors or affecting activation of neurotransmitter receptors. Peripheral BA may also provide the indirect signal to the CNS via farnesoid X receptor (FXR) dependent fibroblast growth factor 15/19 (FGF15/19) pathway or takeda G protein coupled receptor 5 (TGR5) dependent glucagon-like peptide-1 (GLP-1) pathway. Under pathological conditions, alterations in BA metabolites have been discovered as potential pathogenic contributors in multiple neurological disorders. Attractively, hydrophilic ursodeoxycholic acid (UDCA), especially tauroursodeoxycholic acid (TUDCA) can exert neuroprotective roles by attenuating neuroinflammation, apoptosis, oxidative or endoplasmic reticulum stress, which provides promising therapeutic effects for treatment of neurological diseases. This review summarizes recent findings highlighting the metabolism, crosstalk between brain and periphery, and neurological functions of BA to elucidate the important role of BA signaling in the brain under both physiological and pathological conditions.

High altitudes or exposure to hypoxia leads to female reproductive disorders. Circadian clocks are intrinsic time-tracking systems that enable organisms to adapt to the Earth’s 24-h light/dark cycle, which can be entrained by other environmental stimuli to regulate physiological and pathological responses. In this study, we focused on whether ovarian circadian clock proteins were involved in regulating female reproductive dysfunction under hypoxic conditions. Hypobaric hypoxia was found to induce a significantly prolonged estrous cycle in female mice, accompanied by follicular atresia, pituitary/ovarian hormone synthesis disorder, and decreased LHCGR expression in the ovaries. Under the same conditions, the levels of the ovarian circadian clock proteins, CLOCK and BMAL1, were suppressed, whereas E4BP4 levels were upregulated. Results from granulosa cells (GCs) further demonstrated that CLOCK: BMAL1 and E4BP4 function as transcriptional activators and repressors of LHCGR in ovarian GCs, respectively, whose responses were mediated by HIF1ɑ-dependent (E4BP4 upregulation) and ɑ-independent (CLOCK and BMAL1 downregulation) manners. The LHCGR agonist was shown to efficiently recover the impairment of ovulation-related gene ( EREG and PGR ) expression in GCs induced by hypoxia. We conclude that hypoxia exposure causes dysregulation of ovarian circadian clock protein (CLOCK, BMAL1, and E4BP4) expression, which mediates female reproductive dysfunction by impairing LHCGR-dependent signaling events. Adjusting the timing system or recovering the LHCGR level in the ovaries may be helpful in overcoming female reproductive disorders occurring in the highlands.

目的 探讨低温暴露对卵巢组织炎性因子表达的影响及相关分子机制.方法 将6～8周龄雌性小鼠随机分为对照组以及低温暴露组,暴露后取卵巢组织,通过RT?PCR方法检测炎性因子表达水平,Western印迹法检测炎性反应相关蛋白的表达变化及活化情况.体外培养SKOV3细胞,进行温和冷暴露(35℃),通过RT?PCR和Western印迹法检测炎性因子和炎性反应相关蛋白的表达水平及活化情况.结果 低温暴露后卵巢的组织局部出现肿瘤坏死因子(TNF?α)表达水平的显著上调,炎性转录因子NF?κB p65活化.体外培养的SKOV3卵巢癌细胞经温和冷刺激后,同样能观察到TNF?α表达水平的显著上调和炎性转录因子NF?κB p65的诱导活化;在SKOV3细胞中转染NF?κB p65 siRNA后,可显著抑制TNF?α的诱导表达.结论 低温暴露通过诱导卵巢组织NF?κB p65活化介导炎性因子TNF?α上调及炎症反应.

目的 探讨生物钟基因per2对72 h睡眠剥夺(SD)前后脾淋巴细胞功能的影响.方法 将C57小鼠随机分为野生型小鼠对照组(WT)和野生型小鼠睡眠剥夺组(WT-SD);per2基因敲除小鼠(per2-/-)随机分为per2基因敲除小鼠对照组(per2-/-)和per2基因敲除小鼠睡眠剥夺组(per2-/--SD).SD组小鼠使用睡眠剥夺仪进行连续睡眠剥夺72 h.小鼠麻醉后摘脾,使用淋巴细胞分离液分离获得淋巴细胞悬液,利用流式细胞术检测各组别CD4+T、CD8+T、CD3+T以及B220+B比例变化,并通过脂多糖(LPS)刺激3 d体外培养进行流式染色检测上述各淋巴细胞的比例变化,同时使用CCK-8法检测各组小鼠脾B细胞的增殖情况.结果 与WT组小鼠相比,per2-/-组小鼠脾CD4+T、CD8+T以及B220+B细胞比例无明显改变;在进一步的睡眠剥夺实验中发现,per2基因敲除影响T细胞,尤其是CD8+T的组成变化.在LPS刺激实验中,与WT组相比,SD组小鼠脾淋巴细胞增殖更加明显,且与WT-SD组相比,per2-/--SD组B220+B细胞比例增加.同样睡眠剥夺应激条件下,小鼠脾淋巴细胞增殖无明显变化,但与WT-SD组相比,per2-/--SD组脾B细胞降低同时对应T细胞比例升高.结论 per2对脾淋巴细胞的组成影响较小,但在一定程度上参与调节睡眠剥夺后B淋巴细胞对LPS的增殖反应.

目的 探讨急性睡眠剥夺(SD)应激条件下大鼠肝组织胆汁酸转运体昼夜振荡表达的变化规律.方法将大鼠随机分为对照组和实验组.实验组大鼠使用SD仪在24 h昼夜周期内进行急性SD.分离血清后,利用胆汁酸检测试剂盒分析血清中总胆汁酸水平;取肝组织,加入TRIzol试剂后利用组织匀浆仪裂解细胞,通过RT-PCR方法检测肝组织中胆汁酸转运体振荡表达变化.结果与对照组相比,在SD 16 h后总胆汁酸水平显著升高.SD应激条件下,肝组织中胆汁酸转运体钠离子-牛磺胆酸协同转运蛋白(NTCP)和有机阴离子转运多肽(OATP)昼夜振荡变化规律消失,表达水平呈现降低变化;胆汁酸转运体胆盐输出泵(BSEP)和多药耐药相关蛋白2(MRP2)昼夜振荡表达受SD影响相对较小,但MRP2表达峰值呈现一定降低.结论急性SD应激条件可能通过影响胆汁酸转运体NTCP和OATP的振荡表达导致血清总胆汁酸水平异常升高.

目的 研究氧化应激对砷化物诱导肝癌细胞HepG2凋亡反应的影响及TP53诱导糖酵解与凋亡调节因子(TIGAR)对该过程的调节作用机制.方法 比色法检测砷化物刺激作用下肝癌细胞HepG2和正常肝细胞HL7702中的活性氧(ROS)聚集水平;台盼蓝染色法检测砷化物刺激诱导HepG2细胞和HL7702细胞凋亡反应差异;蛋白质免疫印迹和逆转录PCR实验检测砷化物刺激对TIGAR mRNA和蛋白质表达水平的影响;在HepG2细胞中敲低TIGAR表达水平观察其对砷化物诱导ROS聚集反应的影响.结果 砷化物刺激能上调HepG2细胞ROS水平和促进凋亡反应的发生;抑制ROS水平能显著抑制砷化物诱导的细胞凋亡反应;砷化物刺激能诱导TIGAR转录和翻译水平升高;敲低TIGAR能显著上调砷化物诱导的ROS聚集水平和凋亡反应强度.结论 TIGAR通过抑制氧化应激反应拮抗砷化物诱导的HepG2细胞凋亡效应.

目的 探讨大气细颗粒物(PM2.5)暴露后人肺(支气管)上皮Beas-2B细胞中血管内皮生长因子(VEGF)表达水平升高的调控机制.方法 利用生物信息学软件预测人vegf启动子区的潜在转录因子结合位点.采用不同浓度PM2.5(0,12.5,25,50,100μg/ml)刺激Beas-2B细胞,Western印迹法检测VEGF表达调控相关转录因子及其上游蛋白激酶的表达或活化水平.利用相关的siRNA和化学抑制剂分别抑制各信号蛋白的表达和活化后,利用ELISA、Western印迹、RT-PCR和双荧光素报告基因分析法检测PM2.5诱导VEGF表达水平改变情况.结果 发现人vegf启动子区存在转录因子早期生长应答蛋白1(Egr-1)结合位点.经不同浓度PM2.5刺激Beas-2B细胞后,可检测到Egr-1的显著诱导表达.采用特异性siRNA抑制Egr-1表达后,VEGF的转录和蛋白合成反应显著降低,分泌表达水平显著抑制.Egr-1上游蛋白激酶丝裂原活化蛋白激酶p38(p38K)活化水平随PM2.5剂量升高而增强.其特异性抑制剂SB203580预处理Beas-2B细胞后,p38K活化被抑制,Egr-1诱导表达水平显著降低,同时VEGF的转录和蛋白表达均明显受抑.结论 PM2.5可通过诱导Beas-2B细胞中p38K活化引发转录因子Egr-1表达进而上调VEGF的分泌表达水平.

The generation of large numbers of plasma cells (PCs) is a main factor in systemic lupus erythematosus (SLE). We hypothesize that Hspa13, a member of the heat shock protein family, plays a critical role in the control of PC differentiation. To test the hypothesis, we used lipopolysaccharide (LPS)-activated B cells and a newly established mouse line with a CD19cre-mediated, B cell–specific deletion of Hspa13: Hspa13 cKO mice. We found that Hspa13 mRNA was increased in PCs from atacicept-treated lupus-prone mice and in LPS-stimulated plasmablasts (PBs) and PCs. A critical finding was that PBs and PCs [but not naïve B cells and germinal center (GC) B cells] expressed high levels of Hspa13. In contrast, the Hspa13 cKO mice had a reduction in BPs, PCs, and antibodies induced in vitro by LPS and in vivo by sheep red blood cells (SRCs)- or 4-hydroxy-3-nitrophenylacetyl (NP)-immunization. Accordingly, the Hspa13 cKO mice had reduced class-switched and somatically hypermutated antibodies with defective affinity maturation. Our work also showed that Hspa13 interacts with proteins (e.g., Bcap31) in the endoplasmic reticulum (ER) to positively regulate protein transport from the ER to the cytosol. Importantly, Hspa13 mRNA was increased in B220+ cells from patients with multiple myeloma (MM) or SLE, whereas Hspa13 cKO led to reduced autoantibodies and proteinuria in both pristane-induced lupus and lupus-prone MRL/lpr mouse models. Collectively, our data suggest that Hspa13 is critical for PC development and may be a new target for eliminating pathologic PCs.

Objective To investigate the effects of 72 h sleep deprivation(SD)on circadian clock gene expression in the rat liver.Methods Twelve rats were randomly divided into control group and SD group.An SD instrument was used to deprive the rats′sleep for 72 h.Then the abdominal cavity was exposed to obtain liver,and the expression of clock genes was detected by RT-PCR and Western blotting analysis, respectively.Results Compared with the control group, the mRNA levels of clock,npas2 and rev-erbαstrikingly decreased in the livers of the SD group rats.However,per1,per2 and rorαmRNA levels obviously increased.bmal1 and cry1 mRNA expression hardly changed in the control and SD groups. Meanwhile,the protein levels of liver BMAL1,CLOCK,NPAS2,CRY1 and REV-ERBαwere significantly down-regulated and PER1,PER2 and RORαprotein levels were up-regulated in SD group compared with control group.Conclusion 72 h SD can result in abnormal expressions of several circadian clock genes in the rat liver at both transcriptional and translational levels.

Regulatory B cells (Bregs) are a functionally defined B cell subset, and IL-10 is crucial for the suppressive functions of Bregs. However, little is known regarding how IL-10 production is regulated in B cells. To explore the mechanisms by which IL-10 is regulated in B cells, we used mRNA microarrays to screen for molecules that are upregulated in IL-10-producing B cells and identified RNA-binding motif protein 47 ( Rbm47 ) as a post-transcriptional regulator. Rbm47 was found to promote IL-10 production in B cells. We found that Rbm47 promotes the stability of IL-10 mRNA by binding to AU-rich elements in the 3′ untranslated region of Il10 mRNA. In addition, we demonstrated that the overexpression of Rbm47 enabled B cells to facilitate Foxp3 + regulator T-cell induction and reduce the severity of DSS-induced ulcerative colitis. Taken together, these results suggest that Rbm47 plays an important role in regulating IL-10 at the post-transcriptional level, thus promoting the regulatory functions of B cells. The findings presented in this study not only increase our understanding of the post-translational regulation of IL-10 in B cells but also identify a novel strategy for the potential application of Bregs.

Objective To investigate the effect of 36 h continuous sleep deprivation(SD) on circadian clock gene expression in the rat liver and kidney and the alteration of urine biomarker levels.Methods Twelve rats were randomly divided into control group and SD group.An SD device was used to deprive the rats of sleep.After 36 h continuous SD, the abdominal cavity was exposed to obtain livers and kidneys, and RT-PCR and Western blotting were used to detect expression of clock genes.Then,the pelvic cavity was exposed to obtain urine, and the changes in bio-marker total bile acids(TBA) were tested with ELISA.Results Compared with the control group, the mRNA level of liver clock and bmal1 was obviously reduced in the SD-treated rats (P<0.05).However, no obvious change was found in the samples from the kidney.Sharp down-regulation of CLOCK and BMAL1 protein expression was also observed in the rat liver after SD treatment.Urine TBA content in SD treated rats was raised obviously (P<0.001), compared with control.Conclusion Thirty-six hours of continuous SD could result in deregulation of circadian clock gene and cholesterol metabolism disorder in the rat liver.TBA might be used as a noninvasive biomarker of liver injury under SD stress conditions.

Circadian rhythms are driven by the transcriptional and translational oscillator and post-transcriptional modification on clock genes (bmall,clock,cry and per) and clock control genes (rev-erbα,rorα,dbp,tefand hlf),which keeps the body's physiologies,behavior,and other life activities and shows roughly 24-hour oscillation.The circadian clock system consists of the central and peripheral clock system,rhythm input and output system.The input system receives and transfers the light signals to the central clock system that functions as the main pacemaker to generate and output the circadian signals to the peripheral organ,which works together with the local endogenous circadian system to maintain the body's physiological activities.A variety of endogenous and exogenous factors,such as light,temperature,time of food intake,nutrients and metabolic related factors,play important roles in regulating the circadian clock system and maintaining homeostasis of circadian rhythms.Here,we focuses on the regulating system and related regulating factors for the generation and maintenance of circadian rhythms.

Previous studies have shown that under normal physiological conditions thymic B cells play a critical function in T cell negative selection. We tested the effect of thymic B cells on thymic T-cell differentiation in autoimmune diseases including systemic lupus erythematosus (SLE). We found that thymic B cells and CD8- CD4+ and CD4-CD8+T cells increased, whereas CD4+CD8+T cells decreased in lupus-prone mice. Once B cells were reduced, the change was reversed. Furthermore, we found that B cells blocked thymic immature single positive (ISP) CD4-CD8+CD3lo/-RORγt- T cells progression into CD4+CD8+T cells. Interestingly, we found a novel population of thymic immature T cells (CD4-CD8+CD3loRORγt+) that were induced into mature CD4-CD8+CD3+RORγt+T cells by B cells in lupus-prone mice. Importantly, we found that IgG, produced by thymic B cells, played a critical role in the differentiation of thymic CD8+ISP and mature RORγt+CD8+ T cells in lupus-prone mice. In conclusion, B cells blocked the differentiation from thymic CD8+ISP and induced the differentiation of a novel immature CD4-CD8+CD3loRORγt+T cells into mature RORγt+CD8+ T cells by secreting IgG antibody in lupus-prone mice.

The circadian rhythm generated by the endogenous circadian clock system depends on the feedback loop composed of a series of core clock genes (bmal1, clock, cry and per) and clock controlled genes (rev-erbα, rorα, dbp, tef and hlf) .The circadian rhythm is closely related to the body metabolism , which plays important roles in the metabolic process of the major nutrients , such as glucose and fat .Circadian rhythm disorders caused by exogenous factors will significantly increase the risk of metabolic syndrome .This paper focuses on the effect of circadian rhythms on metabolism and their roles in regulating metabolic syndrome .

Circadian rhythms are endogenous 24 h variations found in virtually all physiological processes and behaviors, which are controlled by the transcriptional translational oscillator that consists of a series of core clock genes (bmal1, clock, cry and per) and clock controlled genes (rev-erbα, rorα, dbp, tef and hlf).Clock genes exist in immune organs, tissues and cells, leading to the immune cell function (migration and chemotaxis, phagocytosis, cytotoxicity and so on) and a variety of immune parameters (factor level of circulating immune cells and subsets of the relative and absolute number of cells) showing circadian rhythm changes, and playing an important role in maintaining immune homeostasis.In addition, some immune related diseases are closely correlated with circadian rhythms abnormalities.This paper will focus on the effect of circadian rhythms on immune functions and their roles in some immune related diseases.

Objective To explore whether PERK is involved in the regulation of arsenite-induced autophagy.Methods Human hepatoma cells HepG2 were cultured and treated with arsenite.The expression level of autophagic hallmarks and the activation status of PERK were detected by Western blotting.The transactivation of p53 and the induction of its downstream target genes expression were also detected by Western blotting after knockdown of PERK expression.Transactivity of p53 was detected by dual luciferase reporter assay after knockdown of PERK expression.Results An increase in the LC3BII:I ratio,the induction of Beclin-1 expression and the degradation of p62 were readily observed in arsenite-treated HepG2 cells,but the effects were abolished after knockdown of PERK expression.Furthermore,phosphorylation of p53 at Ser15 and Ser392,transactivation of p53 and the induction of its downstream target gene DAPK1 expression were effectively inhibited under the same PERK knockdown conditions.Conclusion PERK regulates arsenite-induced autophagy by activating p53-dependent DAPK1 upregulation.

目的:探讨Sestrin2在砷化物诱导细胞凋亡反应中的作用及机制.方法:体外培养人肝癌细胞HepG2,以砷化物为刺激源,用免疫印迹和RT-PCR方法检测Sestrin2在砷化物刺激HepG2细胞前后的表达水平差异;用流式细胞术检测敲低Sestrin2前后以及抑制氧化应激反应前后细胞凋亡水平变化情况;用活性氧(ROS)检测试剂盒分析敲低Sestrin2前后细胞在砷化物刺激后的氧化应激水平变化情况.结果:砷化物刺激HepG2细胞后Sestrin2表达水平显著上调;敲低HepG2细胞中Sestrin2表达水平后,细胞凋亡水平明显升高,说明Sestrin2的诱导表达是砷化物诱导细胞凋亡反应中的保护性事件;在敲低Sestrin2表达水平后,砷化物诱导的ROS产生效应和氧化应激反应程度明显加剧;抗氧化剂NAC能够显著逆转Sestrin2对细胞凋亡的保护性效应.结论:Sestrin2在砷化物诱导细胞凋亡反应中可通过抑制ROS产生而发挥拮抗细胞凋亡的保护性作用.

Recently B-cell activating factor (BAFF) was identified by our group and others as a novel therapeutic target for the treatment of autoimmune diseases. To expand upon this, we utilized microarrays to screen for molecules upregulated in B cells from BAFF-inhibited mice with lupus-like disease and identified metabotropic glutamate receptor 3 (Grm3). In addition to confirming the expression of this receptor in B cells, a synthetic agonist of Grm3 was found to downregulate B cells and ameliorate autoimmune symptoms in mice. Conversely, a Grm3 antagonist increased B-cell numbers and further aggravated disease. Thus, these results suggest that activation of Grm3 ameliorates lupus-like disease in mice by reducing B cell numbers. Not only do the findings presented in this study increase our understanding of the inhibitory signals initiated on the surface of B cells, but they also identify a novel potential target for the treatment of autoimmune diseases.