High precision 3 × 3 coupler demodulation algorithm with an additional phase shift judgment module.

Ellipse fitting algorithms (EFAs) have been widely used in 3×3 coupler demodulation systems to reduce the requirement for symmetry of the 3×3 couplers. Based on the relative stability of the splitting ratio and phase difference after the establishment of the 3×3 coupler demodulation system, we solve the problem that EFA fails to work when the stimulating signal is small. Depending on the existence of a symmetry point about the origin, an additional phase shift judgment module is used to determine whether the Lissajous figure is larger than π rad. If the elliptical arc exceeds π rad, the EFA is executed. Otherwise, the previous parameters are used to correct the ellipse. Parameters are updated in real time to ensure high precision. The experimental results show that the total harmonic distortion (THD) of the ameliorated algorithm is improved by 1.28% compared to the EFA without the judgment module with a stimulus amplitude of 30 mV. The proposed scheme can effectively improve the dynamic range of the 3×3 coupler demodulation to reach 125.64 dB @ 1 kHz and 1% THD. The algorithm ensures the effective operation of the EFA under small phase shift conditions and improves the accuracy of phase demodulation.

Iron parameters in pregnant women with beta-thalassaemia minor combined with iron deficiency anaemia compared to pregnant women with iron deficiency anaemia alone demonstrate the safety of iron supplementation in beta-thalassaemia minor during pregnancy.

In patients with beta-thalassaemia intermedia or major, hepcidin induces iron overload by continuously promoting iron absorption. There have been no studies in pregnant women with beta-thalassaemia minor combined with iron deficiency anaemia (IDA), examining whether hepcidin is inhibited by GDF15, as may occur in patients with beta-thalassaemia intermedia or major, or whether the iron metabolism characteristics and the effect of iron supplementation are consistent with simple IDA in pregnancy. We compared and analysed routine blood parameters, iron metabolism parameters, the GDF15 levels, and the hepcidin levels among four groups, namely the beta-thalassaemia (ß) + IDA, ß, IDA, and normal groups. In addition, the ß + IDA and IDA groups received iron supplementation for four weeks. We found no statistically significant correlation between hepcidin and GDF15 in any group, but a positive correlation was observed between hepcidin and ferritin. After iron supplementation, the routine blood parameters and iron metabolism parameters in the ß + IDA group were improved, and the hepcidin content was significantly increased. These results suggest that in pregnant women with beta-thalassaemia minor, hepcidin functions normally to maintain iron homeostasis, and that iron supplementation is effective and safe.

[Effect of S100 calcium binding protein A12 on the pathogenesis of preeclampsia].

OBJECTIVE: To investigate the role of S100 calcium binding protein A12 (S100A12) in the pathogenesis of preeclampsia. METHODS: Sixty patients with preeclampsia were recruited from March 2013 to December 2013 in the First Affiliated Hospital of Zhengzhou University. Among them, thirty cases were defined as the mild preeclampsia group and thirty cases were defined as the severe preeclampsia group. The other thirty healthy pregnant women were recruited in the healthy pregnant women group. The levels of S100A12 protein in maternal peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry of streptavidin peroxidase biotin (SP) method was used to measure the protein expression of S100A12. The trophoblast cells were cultured in vitro with plasma from the three groups, and a blank control group was set up as well. Transwell was used to detect the cytotrophoblast invasion ability. Western blot was used to measure the protein expression level of receptor for advanced glycation end products (RAGE). RESULTS: (1) The levels of S100A12 in maternal peripheral blood of patients with preeclampisa [mild group: (30.8±2.7) µg/L, severe group: (49.3±4.1) µg/L] were significantly higher than that of the control group [(15.8±1.4) µg/L]. In addition, compared with the mild preeclampsia group, the level of S100A12 in the severe preeclampsia group was significantly higher (P < 0.05). (2) Positive immunostaining of S100A12 was observed in the cytoplasm of cytotrophoblast, decidual cells and the placentas from the three groups. The positive rate in the mild preeclampisa group was 77% (23/30); in the severe preeclampsia group it was 93% (28/30); and in the healthy pregnant women group it was 23% (7/30). The positive rates of placenta in the mild and severe preeclampsia groups were significantly higher than that in the healthy pregnant women group (P < 0.05). In addition, compared with the mild preeclampsia group, the positive rate of immunostaining of S100A12 in the severe group was significantly higher (P < 0.05). (3) Cytotrophoblast invasion ability and the expression of RAGE in the mild preeclampsia group were 29.1±3.2 and 0.479±0.038, respectively; in the severe preeclampsia group they were 16.8±2.5 and 0.652±0.059; in the healthy pregnant women group they were 38.6±24.3 and 0.327±0.024; and in the blank control group they were 42.6±5.6 and 0.194±0.011. Cytotrophoblast invasion ability and the expression of RAGE protein in the mild and severe preeclampsia groups were significantly higher than those in the healthy pregnant women group and the control group(P < 0.05). CONCLUSIONS: The expression of S100A12 increased in materal peripheral blood and placenta, and the receptor protein of S100A12 RAGE also had high expression. It suggested that the S100A12 may have some effect on the pathogenesis of preeclampsia.

Bortezomib inhibits hepatocellular carcinoma via the Hippo-Yes-associated protein signalling pathway.

Hepatocellular carcinoma (HCC) is one of the principle causes of cancer-associated death throughout the world. However, the patients with HCC are insensitive to traditional drugs and lack effective therapeutic drugs. Dysregulation of Hippo-Yes-associated protein (YAP) signalling is closely associated with HCC. Bortezomib (BTZ) is mainly used in clinical multiple myeloma. It has recently been confirmed that BTZ could suppress cell proliferation in many different types of cancer. Nevertheless, the precise effects of BTZ on HCC and its possible interactions with the Hippo-YAP signalling pathway in HCC cells remain largely unknown. In this study, HCC cell lines (HepG2 and Huh7) and nude mice with xenograft tumours were used to evaluate the influences of BTZ. Furthermore, we focused on exploring whether BTZ exerts its anti-HCC effect through the Hippo-YAP signalling pathway and aimed to lay a theoretical foundation for BTZ as a potential therapeutic drug for HCC. Herein, our results disclose a new mechanism of BTZ in controlling the cell growth of HCC. BTZ downregulates the level of YAP by promoting LATS1 expression to inhibit the growth of HCC cells, which leads to the phosphorylation of YAP and limits YAP nuclear translocation. In sum, our data confirmed that the Hippo-YAP signalling pathway mediates the anti-HCC effects of BTZ.

EuHD1 protects against inflammatory injury driven by NLRP3 inflammasome.

Non-steroidal anti-inflammatory drugs (NSAIDs) possessing anti-inflammatory, analgesic and antipyretic activities, are widely used in the treatment of osteoarthritis, rheumatism and rheumatoid arthritis. However, its long-term or large use will cause serious gastrointestinal injury or cardiovascular adverse reactions, which limits its clinical application. We have synthesized a new class of NSAIDs, EuHD1, which can release hydrogen sulfide and have better gastrointestinal safety. However, the anti-inflammatory molecular mechanism of the drug is still unclear. In this paper, we explored the mechanism of EuHD1 on NLRP3 inflammasome and its effects on acute lung injury and acute liver injury in mice. In vitro results demonstrated that EuHD1 inhibited macrophage pyroptosis and LDH release induced by LPS combined with ATP. In addition, EuHD1 blocked NLRP3 inflammasome activation and suppressed following Caspase-1 activation and secretion of mature IL-1ß. EuHD1 restrained intracellular ROS production and the formation of ASC oligomers, which inhibited the assembly and activation of NLRP3 inflammasome. In vivo results further showed that EuHD1 alleviated LPS-induced acute lung injury in mice, and inhibited the production of mature IL-1ß and Caspase-1 (p20). Besides, EuHD1 improved D-GalN/LPS-induced acute liver injury, and inhibited SOD/MDA levels and oxidative stress injury, and blocked the activation of NLRP3 inflammasome. In summary, we found that EuHD1 inhibits the assembly and activation of NLRP3 inflammasome through restraining the production of ROS and the formation of ASC oligomers, and has therapeutic effects on acute lung injury and liver injury in mice, indicating that EuHD1 has the potential to treat NLRP3 inflammasome-related diseases.

P2X2 receptors in pyramidal neurons are critical for regulating vulnerability to chronic stress.

Rationale: Stress is a major risk factor for the development of depression. However, the underlying molecular mechanisms of stress vulnerability in depression are largely uncharacterized. Methods: P2X2 receptors (a major receptor for gliotransmitter-ATP) in the medial prefrontal cortex (mPFC) were identified by real-time qPCR, western blots and RNAscope in situ hybridization in chronic social defeat stress model (CSDS). We generated P2X2 conditional knockout mice and overexpressed AAV-P2X2 in CamkIIα-Cre mice. The depression-like behaviors were assessed via CSDS, subthreshold social defeat stress (SSDS), social interaction test (SI), forced interaction test (FIT), forced swimming test (FST), sucrose preference test (SPT), novel stressed feeding (NSF) and open field test (OFT). The neuronal activity and synapse function of P2X2 receptors in the mPFC were detected by in vivo fiber-photometry, patch-clamp techniques and neuronal morphometric analysis. Results: We identified that P2X2 receptors were increased in the mPFC of susceptible mice in CSDS. Conditional knockout of P2X2 receptors in pyramidal neurons promoted resilience of chronic stress-induced depressive-like behaviors, whereas pyramidal neurons - specific gain of P2X2 in the mPFC increased vulnerability to depressive-like behaviors. In vivo fiber-photometry, electrophysiology and neuronal morphometric analysis showed P2X2 receptors regulated neuronal activity and synapse function in the mPFC. Conclusions: Overall, our studies reveal a critical role of P2X2 in mediating vulnerability to chronic stress and identify P2X2 as a potential therapeutic target for treatment of stress-related mood disorders.

M378 exhibits anti-inflammatory activities through NLRP3 signaling pathway.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are frequently used drugs due to their values in attenuating pain, fever and inflammation. Unfortunately, conspicuous adverse effects, such as gastrointestinal (GI) damage and/or cardiovascular events have impeded their application in clinic. M378 is a novel hydrogen sulfide-releasing NSAIDs with uncompromised potency and negligible toxicity compared to the existing NSAIDs. However, its anti-inflammatory activity and mechanism are still an enigma. Here we investigated the effect of M378 on the NLRP3 inflammasome signaling pathway and addressed the underlying molecular mechanism. Our data in vitro showed that M378 dose-dependently inhibited the cleavage of Caspase-1 and the secretion of active IL-1ß and blocked NLRP3-dependent pyroptosis in LPS-primed J774A.1 macrophages. Furthermore, M378 remarkably inhibited upstream ASC oligomerization and ROS production regarding the process of NLRP3 inflammasome assembly. Our data in vivo demonstrated that M378 protected mice from acute liver injury, reducing the levels of ALT/AST and IL-1ß and improving hepatic pathological damages. Immunoblot analysis revealed that M378 inhibited the expressions of Caspase-1 and IL-1ß in liver tissues of ALI mice. We also showed that M378 alleviated IL-1ß secretion and peritoneal neutrophils infiltration in MSU-elicited acute peritonitis mice. In conclusion, M378 exerted its anti-inflammatory effect both in vitro and in vivo and its mechanisms are at least connected to its inhibitory performance on the generation of ASC oligomers and ROS production. These findings give an insight. into the molecular mechanism of hydrogen sulfide-releasing NSAIDs and support a potent therapeutic role of M378 in the treatment of NLRP3-driven inflammatory diseases.