Objective To investigate the change of adiponectin level with menopause status in women aged 40 to 65, and its relationship with androgen. Methods A cross-sectional study included woman (aged from 40 to 65) who were in hospital for routine check-up at the Sun Yat-sen Memorial Hospital from August to October in 2013. All subjects underwent laboratory examinations of adiponectin, sex hormone binding globulin (SHBG), dehydroepiandrosterone-sulfate (DHEA-S), total testosterone (TT), collected anthropometric measurements and then calculated free androgen index (FAI) and body mass index (BMI). According to their menstrual status, the subjects were divided into 4 groups: premenopausal group with 119 subjects, perimenopausal group with 60 subjects, early postmenopausal group with 62 subjects, late postmenopausal group with 64 subjects. Results (1) Adiponectin levels declined to its lowest level in menopausal transition and gradually becoming higher after menopause, which showed a U-shaped trajectory. When compared adiponectin levels in late postmenopausal group [(13 ± 5) mg/L] with those in perimenopausal [(8 ± 6) mg/L] or early postmenopausal group [(9 ± 6) mg/L], it all showed significantly difference (P<0.05). (2) Both the adiponectin levels were negatively correlated with waistline in the 4 groups (premenopausal group, r=-0.276;perimenopausal group, r=-0.334;early postmenopausal group, r=-0.211;late postmenopausal group, r=-0.218; all P<0.05). Levels of adiponectin were positively correlated with SHBG (r=0.536, P<0.05) and negatively with FAI (r=-0.363, P<0.05) in menopausal transition, while in late postmenopausal group, negatively correlated with level of DHEA-S (r=-0.450, P<0.05). When adjusted for age, BMI and waistline, the above correlations still exist. Conclusions Adiponectin levels declined to its lowest level in menopausal transition and gradually becoming higher after menopause, which showed a U-shaped trajectory during the sequential menopause status transition in middle aged women. Low level of adiponectin in menopausal transition is closely associated with the relative excess androgen occurred during this stage.

Objective:To investigate whether miR-548b-3p can regulate the malignant biological traits of hepatocellular carcinoma cells by down-regulating the expression of decoy receptor 3( DcR3). Methods:The miR-548b-3p levels in 5 common hepatoma cell lines Hep3b, HepG2, Huh7.5.1, PLC and 97H were detected by real-time fluorescence quantitative PCR. The experimental hepatoma cell lines were purchased from Shanghai Cancer Institute. The subcutaneous tumor formation experiment in nude mice included two parts: miR-548b-3p over expression group and down expression group. miR-548b-3p over expression experiment included Huh7.5.1- miR-548b-3p cell group and Huh7.5.1-NC cell group. The experimental group with low expression of miR-548b-3p included PLC- miR-548b-3p-sponge group and PLC-NC cell group, with 5 nude mice in each group. The nude mice used in the experiment were male Balb/c strain, body weight 20-25 g, 4-6 weeks old. Starbase software was used to predict whether there were binding sites between miR-548b-3p and DcR3. Dual luciferase reporting assay to verify whether DcR3 is the target gene of miR-548b-3p. Rescue experiments were conducted to verify whether the effects of miR-548b-3p on the proliferation, invasion and migration of liver cancer cells were realized through DcR3. Statistical analysis was performed using Graphpad Prism 9. Measurement data with normal distribution were expressed as mean±standard deviation( ± s), and t-test was adopted for comparison between general measurement data groups. Results:The relative expression level of 2 -ΔΔCt was used as the parameter for real-time fluorescence quantitative PCR comparison. Hep3b expression level was 1, HepG2 expression level was 0.902, Huh7.5.1 expression level was 0.712, PLC expression level was 1.293, and 97H expression level was 0.818. The final results showed that, The highest expression of miR-548b-3p was in PLC cells, and the lowest expression was in Huh7.5.1 cells. The subcutaneous tumor formation experiment of nude mice showed that after 4 weeks, the tumor volume of Huh7.5.1- miR-548b-3p was(444.77±142.34) mm 3, and that of Huh7.5.1-NC was(918.80±139.21) mm 3. The tumor volume of PLC- miR-548b-3p-sponge was(407.49±58.50) mm 3, and that of PLC-NC was(218.62±47.55) mm 3. The binding sites of miR-548b-3p and DcR3 were predicted by Starbase software. Dual luciferase assay showed that DcR3 was the target gene of miR-548b-3p. Rescue experiments verified that the effects of miR-548b-3p on the proliferation, invasion and migration of liver cancer cells were realized through DcR3. Conclusion:miR-548b-3p can regulate the proliferation, invasion, migration and other malignant biological traits of liver cancer cells, and it is achieved by down-regulating the expression level of DcR3.

Objective:To explore the relationship between amniotic fluid and peripheral blood inflammatory factors and the pregnancy outcomes after emergency cervical cerclage, and to identify effective indicators for predicting adverse pregnancy outcomes after the procedure.Methods:A case-control study was conducted, including pregnant women who were hospitalized at Sun Yat-sen Memorial Hospital, from January 1, 2013, to July 31, 2019, and underwent emergency cervical cerclage due to cervical dilatation at gestational age between 16 and 28 weeks. A total of 85 pregnant women who underwent amniocentesis for the detection of amniotic fluid inflammatory factors during the perioperative period were included. Based on whether their baby was perinatal death, the participants were divided into the case group (28 cases with perinatal death) and the control group (57 cases with live births). Univariate logistic regression analysis was performed to identify risk factors associated with adverse pregnancy outcomes, followed by multivariate logistic regression analysis to establish a regression model and nomogram.Results:(1) The levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6, IL-8, IL-10 in the amniotic fluid during the perioperative period and postoperative serum C-reactive protein (CRP) were significantly higher in the case group compared to the control group (all P<0.05). The case group underwent emergency cervical cerclage at an earlier gestational age compared to the control group, and their cervical dilation was greater than that of the control group (all P<0.05). However, there were no significant differences in the white blood cell counts, neutrophil percentage, and the level of preoperative CRP in the peripheral blood of pregnant women during the perioperative period (all P>0.05). (2) Univariate logistic regression analysis showed that the levels of amniotic fluid WBC, TNF-α, IL-1β, IL-2 receptor (IL-2R), IL-6, IL-8, IL-10, postoperative CRP in the peripheral blood, gestational age at cerclage and cervical dilation were associated with adverse pregnancy outcomes (all P<0.05). Multivariate regression analysis indicated that only the levels of amniotic fluid WBC and TNF-α were independent risk factors for perinatal death. (3) Based on clinical practice, a multivariate logistic regression model was constructed including the levels of amniotic fluid TNF-α, WBC, gestational age at cervical cerclage, and cervical dilation. A nomogram and calibration curve were plotted, which suggested its good predictive value for adverse pregnancy outcomes. Conclusions:During the perioperative period of emergency cervical cerclage, the levels of amniotic fluid WBC, TNF-α, IL-1β, IL-2R, IL-6, IL-8, IL-10 are associated with adverse pregnancy outcomes, with amniotic fluid WBC and TNF-α showing the closest relationship. However, there is no significant correlation between maternal peripheral hemogram during the perioperative period and adverse pregnancy outcomes. A model constructed by amniotic fluid TNF-α, WBC, cervical cerclage gestational age, and cervical dilation has a good predictive effect on adverse pregnancy outcomes.

Objective To investigate the effects of fluid shear stress on rolling adhesion of neutrophils on immobilized platelets under flows. Methods Experiments were performed at the parallel plate flow chamber. Platelets were adhered to the functionalized flow chamber bottom which were coated with vWF-A1 first, and then washed with PBS under wall shear stress (WSS) of 1 Pa for different time (0 min, 2.5 min, 7.5 min). A high-speed camera was used to observe and record the rolling adhesion events of neutrophils on immobilized platelets under 50 mPa WSS, and the adhesion parameters such as the number of adhesion events, the tether lifetime of cells and rolling velocity. Results Neutrophils could specifically bind to the immobilized platelets on vWF-A1-coated bottom of the flow chamber. Mechanical stimulation on immobilized platelets had no effects on the tether lifetime of neutrophils on the platelets, but up-regulated the adhesive ratio of neutrophils on the platelets and slowed down the rolling of neutrophils on the platelets. Conclusions Mechanical stimulation on the immobilized platelets will significantly make the circulating neutrophils to be captured easily and promote the rolling adhesion of neutrophils on platelets.

P-selectin engagement of P-selectin glycoprotein ligand-1 (PSGL-1) causes circulating leukocytes to roll on and adhere to the vascular surface, and mediates intracellular calcium flux, a key but unclear event for subsequent arresting firmly at and migrating into the infection or injured tissue. Using a parallel plate flow chamber technique and intracellular calcium ion detector (Fluo-4 AM), the intracellular calcium flux of firmly adhered neutrophils on immobilized P-selectin in the absence of chemokines at various wall shear stresses was investigated here in real time by fluorescence microscopy. The results demonstrated that P-selectin engagement of PSGL-1 induced the intracellular calcium flux of firmly adhered neutrophils in flow, increasing P-selectin concentration enhanced cellular calcium signaling, and, force triggered, enhanced and quickened the cytoplasmic calcium bursting of neutrophils on immobilized P-selectin. This P-selectin-induced calcium signaling should come from intracellular calcium release rather than extracellular calcium influx, and be along the mechano-chemical signal pathway involving the cytoskeleton, moesin and Spleen tyrosine kinase (Syk). These results provide a novel insight into the mechano-chemical regulation mechanism for P-selectin-induced calcium signaling of neutrophils in flow.
Calcium Signaling ; Female ; Humans ; Male ; Membrane Glycoproteins ; metabolism ; Neutrophils ; metabolism ; P-Selectin ; metabolism ; Stress, Mechanical ; Syk Kinase ; metabolism