The association between adipokines and pulmonary diseases: a mendelian randomization study.

BACKGROUND: The role of adipokines in the development of lung diseases is significant, yet their specific relationship with different lung diseases remains unclear. METHODS: In our research, we analyzed genetic variations associated with adipokines and various lung conditions such as interstitial lung disease, chronic obstructive pulmonary disease, asthma, lung cancer, sleep apnea, pneumonia, and tuberculosis, using data from public genome-wide studies. We employed Mendelian randomization techniques, including inverse variance weighting, weighted median, and MR-Egger regression methods, and conducted sensitivity checks to validate our findings. RESULTS: A study using the FinnGen database, which included 198,955 participants, identified 13 SNPs associated with adiponectin. Notably, adiponectin was found to significantly reduce the risk of interstitial lung disease and idiopathic pulmonary fibrosis. However, little evidence was found to establish a direct cause-effect relationship between the six adipokines and several other lung conditions, including sarcoidosis, asthma, chronic obstructive pulmonary disease, lung cancer, tuberculosis, pneumonia, and sleep apnea syndrome. CONCLUSION: This study reveals a reverse link between adiponectin levels and the likelihood of interstitial lung disease, including idiopathic pulmonary fibrosis.

METTL3-mediated lncRNA HOXD-AS1 stability regulates inflammation, and the migration and invasion of trophoblast cells via the miR-135a/ ß-TRCP axis.

Background: Preeclampsia (PE) is a serious pregnancy-specific syndrome associated with the inadequate invasion of trophoblast cells and inflammation of the uterus. A previous study found that lncRNA HOXD-AS1 promotes PE. However, its regulatory network requires additional exploration. Methods: HOXD-AS1-targeted miRNAs and genes were predicted by different databases in a bioinformatics analysis. The expression HOXD-AS1 and its potential m6A methylase (METTL3) were detected in placentas from healthy female patients with PE. The targeting relationship and role of the HOXD-AS1/miR-135a/ß-TRCP axis in trophoblast cells were demonstrated by overexpression/knockdown assays. The levels of ß-TRCP downstream pathway proteins IκBα, NF-κB, and p65 were measured. The levels of inflammatory factors in cell supernatants were detected by ELISA. To verify the molecular mechanism of ß-TRCP in PE, IκBα was co-overexpressed in ß-TRCP in trophoblast cells. Results: The levels of METTL3, HOXD-AS1, and ß-TRCP were elevated in PE placental tissues, while miR-135a levels were reduced. MiR-135a was found to be targeted by HOXD-AS1, and HOXD-AS1 expression was maintained at a high level by METTL3-mediated m6A methylation. Overexpression of METTL3, HOXD-AS1, and ß-TRCP, and knockdown of miR-135a in HTR-8/SVneo cells all inhibited cell invasion and migration, and promoted apoptosis and the secretion of inflammatory factors. Knockdown of METTL3, HOXD-AS1, and ß-TRCP, and overexpression of miR-135a had the opposite effects. Furthermore, IκBα expression was negatively associated with ß-TRCP expression, and the levels of NF-κB, p65, and NLRP3 were positively regulated by ß-TRCP. A high level of ß-TRCP expression attenuated the effect of HOXD-AS1 knockdown in trophoblast cells. Conclusion: METTL3 functioned to maintain a high level of HOXD-AS1 expression in PE, which influenced inflammation and the migration and invasion of trophoblast cells via the miR-135a/ß-TRCP axis and NF-κB pathway.

Effect of CMB Carrying PTX and CRISPR/Cas9 on Endometrial Cancer Naked Mouse Model.

Endometrial cancer, one of the most common gynecological cancers in women. Patients with advanced or recurrent disease have poor long-term outcomes. The current experiment explore the roles of cationic microbubbles (CMBs) carrying paclitaxel (PTX) and CRISPR/Cas9 plasmids on the xenotransplantation model of mice with endometrial cancer. The tumor histology, tumor cell viability, cell cycle, and invasion ability were investigated. Meanwhile, the P27, P21, GSK-3, Bcl-2 associated death promoter (Bad), mammalian target of rapamycin (mTOR), and C-erbB-2 expressions were evaluated by qRT-PCR and western blotting, respectively. CMB-PTX-CRISPR/Cas9 had an inhibitory action on the tumor growth, tumor cell viability, cell cycle, and invasion ability of the mouse xenograft model of endometrial cancer. The CMB-PTX-CRISPR/Cas9 increased the GSK-3, P21, P27, and Bad expression levels, while reduced the C-erbB-2 and mTOR expressions. CMBs loaded with both PTX and CRISPR/Cas9 plasmids may be a new combination treatment with much potential. CMB-PTX-CRISPR/Cas9 may regulate the tumor cell viability, invasion, and metastasis of endometrial cancer naked mouse model by upregulating expressions of GSK-3, P21, P27, and Bad.

The relationship between hsa_circ_0051326 and HLA-G expression in the blood of patients with pre-eclampsia.

OBJECTIVES: To investigate the relationship between hsa_circ_0051326 and HLA-G expression in the blood of patients with pre-eclampsia. MATERIAL AND METHODS: Real-time PCR (qRT-PCR) was used to detect the hsa_circ_0051326 expression level. Enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and western blotting were used to detect HLA-G expression in blood in 50 patients with pre-eclampsia and 50 normal pregnant women. RESULTS: HLA-G protein expression level was decreased significantly in the blood of patients with pre-eclampsia. hsa_circ_0051326 and HLA-G mRNA in blood were decreased significantly in the pre-eclampsia patients compared with normal pregnant women. There was a positive correlation between the expression of serum hsa_circ_0051326 with HLA-G mRNA. CONCLUSIONS: Serum hsa_circ_0051326 was a new diagnostic biomarker for pre-eclampsia with equivalent efficiency of HLA-G. Maternal serum hsa_circ_0051326 level may pre-diagnose potentially pre-eclampsia before delivery.

Methylation Status of the miR-141-3p Promoter Regulates miR-141-3p Expression, Inflammasome Formation, and the Invasiveness of HTR-8/SVneo Cells.

MicroRNA-141 (miR-141-3p) is upregulated in preeclampsia. This study investigated the effect of methylation of the miR-141-3p promoter on cell viability, invasion capability, and inflammasomes in vitro. The expression of miR-141-3p and methylation status of the miR-141-3p promoter were examined by RT-qPCR and pyrosequencing in villus tissues of women with spontaneous delivery (VTsd), villus tissues of women with preeclampsia (VTpe), and also in HTR-8/SVneo cells treated with a miR-141-3p inhibitor and 20 µmol/L 5-aza-2'-deoxycytidine (5-Aza), a DNA methyltransferase inhibitor. Cell viability and invasion were evaluated by CCK-8 and transwell assays. In addition, the levels of CXCL12, CXCR4, CXCR2, MMPs, NLRP3, and ASC expression were assessed by western blotting, and IL-1ß and IL-18 concentrations were assayed by ELISA. miR-141-3p expression was upregulated, and the levels of miR-141-3p promoter methylation and CXCL12, CXCR4, and CXCR2 expression were decreased in VTpe relative to VTsd. In HTR-8/SVneo cells, hypomethylation caused by 5-Aza treatment increased miR-141-3p expression, while DNA methyltransferase 3 (DNMT3) transfection decreased miR-141-3p expression. miRNA-141-3p induced NLRP3, IL-1ß, and IL-18 production, decreased CXCR4, MMP, and MMP2 production, and suppressed cell growth and invasion. Furthermore, we observed that NLRP3 plays an important mediatory role in the effects of miR-141-3p described above. Decreased methylation of the miR-141-3p promoter increases miR-141-3p expression, which in turn increases NLRP3 expression, resulting in higher IL-1ß and IL-18 levels and lower levels of MMP2/9 and CXCR4. We conclude that modification of the miR-141-3p promoter might be a curial mediator in preeclampsia.

MicroRNA­524­5p regulates the proliferation and invasion of HTR­8/SVneo trophoblasts by targeting NUMB in the Notch signaling pathway.

Preeclampsia is a pregnancy disorder that is primarily associated with maternal and neonatal or fetal morbidity and mortality. The discovery of dysregulated microRNAs (miRs) and their roles in preeclampsia has provided new insight into the mechanisms involved in pregnancy­related disorders. In the present study, quantitative PCR demonstrated that the expression levels of miR­524­5p were lower in patients with preeclampsia than those in normal pregnant women. Cell Counting Kit­8 and Transwell assays indicated that overexpression of miR­524­5p promoted the proliferation and invasion of HTR­8/SVneo cells, whereas inhibition of miR­524­5p suppressed HTR­8/SVneo cell proliferation and invasion. Furthermore, NUMB endocytic adaptor protein (NUMB), a negative regulator of the Notch signaling pathway and a target gene of miR­524­5p, limited the effects of miR­524­5p on HTR­8/SVneo cell invasion and migration. The present study demonstrated that miR­524­5p regulated the proliferation and invasion of HTR­8/SVneo cells at least partly by targeting NUMB to regulate the Notch signaling pathway.

MiR-135a-5p promotes the migration and invasion of trophoblast cells in preeclampsia by targeting ß-TrCP.

BACKGROUND: MiR-135a-5p is an important regulator of cell migration and invasion in several diseases. However, the biological functions and mechanisms of miR-135a-5p in women with preeclampsia (PE) remain unclear. METHODS: The levels of miR-135a-5p and beta-transducin repeat containing E3 ubiquitin protein ligase (ß-TrCP) expression in samples of placenta tissue from PE patients and healthy control subjects were determined by quantitative real-time PCR. The effects of miR-135a-5p and ß-TrCP on cell migration, invasion, and epithelial-mesenchymal transition (EMT) in two trophoblast cell lines (HTR-8/SVneo and TEV-1) were examined using wound healing, Transwell, and western blot assays, respectively. A luciferase reporter assay was performed to confirm the association between miR-135a-5p and ß-TrCP, and an in vivo mouse model was established and used to analyze the effect of ß-TrCP on PE clinical phenotypes. RESULTS: We found that miR-135a-5p expression was significantly decreased and negatively correlated with ß-TrCP expression in the placental tissues of pregnant women with PE. Cellular function experiments showed that overexpression of miR-135a5p promoted the migration and invasion of trophoblast cells in vitro. Furthermore, ß-TrCP was confirmed as a target gene of miR-135a-5p in trophoblast cells. Notably, overexpression of ß-TrCP significantly reversed the effect of miR-135a-5p on migration and invasion of trophoblast cells. At the molecular level, decreases in E-cadherin levels and increases in N-cadherin, Vimentin, and ß-catenin levels that were induced by miR-135a-5p overexpression were attenuated by ß-TrCP overexpression. CONCLUSIONS: Our findings demonstrate that miR-135a-5p promotes the migration and invasion of trophoblast cells by targeting ß-TrCP.

ß-TrCP suppresses the migration and invasion of trophoblast cells in preeclampsia by down-regulating Snail.

Insufficient trophoblast invasion has been shown to contribute to the occurrence and progression of preeclampsia (PE). Recently, beta-transducin repeat containing E3 ubiquitin protein (ß-TrCP) was shown to function as a ubiquitination regulator in regulating the proliferation and invasion of various cell types. In this study, we employed an in vitro model of trophoblasts to investigate the role played by ß-TrCP in the pathogenesis of PE. The levels of ß-TrCP in newly delivered placentas from 15 pregnant women with PE and 15 healthy pregnant women were detected by quantitative real-time PCR and western blot assays. The effects of ß-TrCP on cell migration, invasion, and epithelial-mesenchymal transition (EMT) in two trophoblast cell lines (HTR-8/SVneo and TEV-1) were examined using wound healing assays, Transwell assays, and western blot assays, respectively. Rescue experiments were performed by treating ß-TrCP knockdown or ß-TrCP expressing trophoblasts with si-Snail transfection or a proteasome inhibitor (MG132). ß-TrCP mRNA and protein expression levels were significantly increased in the PE placentas when compared to the normal control placentas. ß-TrCP overexpression significantly inhibited cell migration and invasion, while silencing of ß-TrCP promoted cell migration and invasion of the two trophoblast cell lines. Furthermore, we demonstrated that ß-TrCP-mediated ubiquitination might inhibit the EMT process of trophoblasts by down-regulating Snail expression. Our results suggest that both ß-TrCP mRNA and protein expression were up-regulated in the PE placentas. ß-TrCP impeded the migration and invasion of trophoblasts by suppressing Snail expression. This implicates the ubiquitin-proteasome pathway in the pathogenesis of PE, and suggests ß-TrCP as a potential target for treating PE.

Dietary patterns and gestational hypertension in nulliparous pregnant Chinese women: A CONSORT report.

It has been well established that dietary patterns play important roles in the pathogenesis and development of hypertension. Our aim was to investigate the association between pregnancy dietary patterns and the risk of hypertension among nulliparous pregnant Chinese women.A cross-sectional, case-control study.Three hospitals in Haikou, the capital of Hainan Province, South China.A total of 2580 participants who reported dietary intake using a validated food frequency questionnaire (FFQ).Four primary dietary patterns were identified by principal component factor analysis and labeled as traditional Chinese, animal food, Western food, and salty snacks patterns. Women with high scores on pattern characterized by salty snacks were at increased risk.This study suggests that dietary pattern characterized by salty snack increases the risk of hypertension during pregnancy.

Hypoxia-induced microRNA-141 regulates trophoblast apoptosis, invasion, and vascularization by blocking CXCL12ß/CXCR2/4 signal transduction.

BACKGROUND: An impaired trophoblast invasion ability contributes to the development of pre-eclampsia (PE), and can be induced by the altered expression of various microRNAs (miRs). MiR-141 and CXCL12ß (C-X-C motif chemokine ligand 12) signaling regulate trophoblast invasion and vascularization capabilities during PE pathogenesis; however, their interactions and underlying mechanisms of action remain unclear. We investigated how miR-141 modulates trophoblast invasion, with a focus on its interaction with CXCL12ß signaling. METHODS: A PE model was established by using HTR-8/SVneo cells, which were first cultured with 2% O2 for 48 h, and then with 5% O2. The expression of miR-141 in human villous trophoblast HTR-8/SVneo cells was modulated with mimics or an inhibitor, and analyzed by quantitative RT-PCR. CXCL12ß levels were determined by ELISA. Cell apoptosis was determined by flow cytometry, and the invasion and vascularization capabilities of trophoblasts were evaluated by Transwell and tube formation assays, respectively. Binding of miR-141 with CXCL12ß mRNA was verified by the dual luciferase assay. Protein levels were estimated by western blotting. RESULTS: MiR-141 expression was significantly induced by hypoxia in HTR-8/SVneo cells. MiR-141 was found to promote apoptosis and inhibit the invasion and vascularization abilities of HTR-8/SVneo cells under conditions of hypoxia. MiR-141 could directly bind with the 3'UTR region of CXCL12ß mRNA and inhibit its translation. In addition, we proved that miR-141 could inhibit the invasion and vascularization abilities, and promote the apoptosis of HTR-8/SVneo cells by targeting CXCL12ß under hypoxic conditions. Furthermore, we demonstrated that arachidonic acid could reverse the invasion and apoptosis abilities of HTR-8/SVneo cells mediated by CXCL12ß during hypoxia. In terms of mechanism, MiR-141 could downregulate MMP2, p62, and LC3B expression, and upregulate ROCK1 and RhoA expression in HTR-8/SVneo cells by targeting the CXCL12ß gene during hypoxia. The effects of CXCL12ßon HTR-8/SVneo cells could be reversed by arachidonic acid (ARA). CONCLUSION: Induction of miR-141 by hypoxia promotes apoptosis, and inhibits the invasion and vascularization capabilities of HTR-8/SVneo cells by suppressing the CXCL12ß and CXCR2/4 signaling pathways.

Inhibition of Tyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase Activation Protein Zeta (YWHAZ) Overcomes Drug Resistance and Tumorigenicity in Ovarian Cancer.

BACKGROUND/AIMS: Cancer stem-like cells are the main cause of tumor occurrence, progression, and therapeutic resistance. However, the precise signals required for the maintenance of the stem-like traits of these cells in ovarian cancer remain elusive. We have thus worked to elucidate the functional role of Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ), a gene encoding the 14-3-3ζ protein, in the regulation of multidrug resistance and stem cell-like traits in ovarian cancer. METHODS: We detected the YWHAZ levels in human ovarian cancer specimens and cell lines using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blots. MTS assays, soft agar colony formation assays, migration assays, cell cycle analysis, sphere formation assays, and flow cytometry were applied to investigate the functional role of YWHAZ in ovarian cancer. RESULTS: Our data reveals substantially increased YWHAZ expression in both cisplatin- and paclitaxel-resistant ovarian cancer cells. Silencing YWHAZ restored the sensitivity of resistant ovarian cancer cells to cisplatin and paclitaxel. Furthermore, in vitro studies showed that down-regulation of YWHAZ inhibited cell cycle progression, migration, and the expression of stem cell markers. Moreover, tumorigenicity was suppressed in tumor-bearing BALB/c nude mice following YWHAZ knockdown. Additionally, we demonstrated that the expression of YWHAZ was directly down-regulated by miR-30e in resistant ovarian cancer cells. CONCLUSION: Our results have led to new insights into the essential role of YWHAZ in the regulation of tumourigenesis, stem-like traits, and drug resistance in ovarian cancer, thereby helping to identify a potential target for ovarian cancer therapy.

Tumor-associated macrophages promote the metastasis of ovarian carcinoma cells by enhancing CXCL16/CXCR6 expression.

This study investigated the underlying mechanism by which C-X-C motif chemokine ligand 16 (CXCL16)/C-X-C motif chemokine receptor 6 (CXCR6) signaling is activated by tumor-associated macrophages and assists in regulating the metastasis of ovarian carcinoma. Specimens of ovarian carcinoma tissue and adjacent tissue were collected from 20 ovarian carcinoma patients. Human THP-1 cells were induced to differentiate into macrophages, which were then co-cultured with SKOV3 cells and low concentrations of tumor necrosis factor-α (TNF-α) to simulate the inflammatory microenvironment of ovarian carcinoma. Additionally, small interfering RNA (siRNA) targeting CXCR6 was transfected into SKOV3 cells; after which, the levels of nuclear factor kappa B p65 (NF-κB p65) protein and phosphorylated PI3K and Akt were measured. The migration and invasion abilities of the SKOV3 cells were also tested. The levels of TNF-α, interluekin-6 (IL-6), NF-κB p65, CXCL16, and CXCR6 expression in the ovarian carcinoma tissues were higher than those in the precancerous tissues. CXCR6 expression was positively correlated with TNF-α, IL-6, and CXCL16 expression. Co-culture of SKOV3 cells with macrophages significantly promoted CXCL16, CXCR6, NF-κB, and p65 expression by the SKOV3 cells, increased their levels of phosphorylated PI3K and Akt, and increased the migration and invasion abilities of SKOV3 cells. Silencing of CXCR6 or blocking the PI3K/Akt signal pathway markedly attenuated the expression of NF-κB p65 and phosphorylation of PI3K and Akt, as well as the migration and invasion abilities of SKOV3 cells. These findings demonstrate that macrophages can promote the migration and invasion of ovarian carcinoma cells by affecting the CXCL16/CXCR6 pathway.

Upregulation of SNHG3 expression associated with poor prognosis and enhances malignant progression of ovarian cancer.

BACKGROUND: Aberrant expression of long non-coding RNAs is involved in the progression of ovarian cancer. However, the clinical significance and biological functions of SNHG3 expression was little known in ovarian cancer (OC). METHODS: The SNHG3 expression in ovarian cancer tissues and paired adjacent normal tissues was detected using quantitative real time polymerase chain reaction (qRT-PCR). Gain-of function and loss-of function assays were performed in ovarian cancer cells to demonstrate the effects of SNHG3 expression on cell proliferation and invasion. The relative protein expression levels were determined using western blot analyses. RESULTS: The expression of SNHG3 was significantly up-regulated in ovarian cancer tissues compared with adjacent normal tissues. Higher SNHG3 expression levels positively associated with FIGO stage, lymph node metastasis, and poor prognosis of ovarian cancer patients. Univariate and multivariate Cox regression analysis implied that FIGO stage, lymph node metastasis, higher SNHG3 expression were independent prognostic factors for overall survival (OS) rate in ovarian cancer patients. Gain-of function and loss-of function assays demonstrated that SNHG3 knockdown inhibited ovarian cancer cell proliferation and invasion abilities. However, SNHG3 overexpression promoted ovarian cancer cell proliferation and invasion abilities. Furthermore, cell proliferation and invasion related protein CyclinD1, CDK1, MMP9 and MMP3 were significantly downregulated after SNHG3 knockdown in ovarian cancer cells, while SNHG3 overexpression had reverse effects. In addition, SNHG3 functioned as an oncogene by regulating GSK3ß/ß-catenin signaling activity in ovarian cancer. CONCLUSIONS: Taken together, our data provide that SNHG3 has potential clinical value of and may serve as target of ovarian cancer treatment.

CXCR2 is decreased in preeclamptic placentas and promotes human trophoblast invasion through the Akt signaling pathway.

INTRODUCTION: CXCR2, the receptor of the CXC chemokines, plays a critical role in cell migration and invasion in many types of cancer. It is unclear what impact CXCR2 may have on Preeclampsia (PE), a pregnancy-specific disease, which is related to insufficient trophoblast invasion. The aim of this study was to investigate the expression pattern of CXCR2 in the placentas of healthy and PE pregnancies, and to investigate the molecular mechanism of CXCR2 involvement in the development of PE. METHODS: CXCR2 expression levels in newly delivered placentas from 38 pregnant women with PE and 21 healthy pregnant women were detected using quantitative real-time PCR, immunohistochemistry and Western blot assays. The effect of CXCR2 on trophoblast invasion and the underlying mechanisms were examined in two trophoblast cell lines (HTR-8/SVneo and TEV-1 cells). RESULTS: CXCR2 mRNA and protein expression levels were significantly decreased in preeclamptic placentas than normal control. The invasive abilities of the two trophoblast cell lines were significantly inhibited when CXCR2 was silenced, but that CXCR2 overexpression promoted trophoblast cells invasion. In addition, silencing CXCR2 reduced the expression of matrix metalloproteinase 2 and 9 (MMP2 and MMP9) and phosphorylated Akt (p-Akt). Furthermore, an Akt inhibitor suppressed the expression of MMP-2 and MMP-9. DISCUSSION: Our results suggest that the decreased CXCR2 may contribute to the development of preeclampsia through impairing trophoblast invasion by down-regulating MMP-2 and MMP-9 via the Akt signaling pathway.

Preeclampsia in twin pregnancies: association with selective intrauterine growth restriction.

OBJECTIVE: To identify the association between preeclampsia (PE) and selective intrauterine growth restriction (sIUGR) in twin pregnancies. METHODS: This was a retrospective cohort study of 1004 twin pregnancies from 2008 to 2014. We specifically compared the incidence, clinical characteristics and outcomes of PE between sIUGR and normal-growth twin pregnancies. RESULTS: PE occurred more frequently in sIUGR pregnancies [29.0% (51/176)] than in normal-growth twin pregnancies [13.1% (99/756), p < 0.001, adjusted odds ratio 3.29]. Among sIUGR, the incidence of PE was significantly higher in dichorionic (DC) pregnancies (37.5%, 30/80) than in monochorionic (MC) pregnancies (21.9%, 21/96). The rates of onset at <32 weeks (p = 0.045) and of severe PE (p = 0.025) were higher in sIUGR pregnancies with PE. The systolic blood pressure was also higher in sIUGR pregnancies with PE (152.6 ± 11.8 mmHg) than in normal-growth pregnancies with PE (148.0 ± 8.2 mmHg) (p = 0.042). Additionally, more sIUGR pregnancies were delivered at 32-36 weeks (p = 0.001), and fewer were delivered at ≥36 weeks (p < 0.001). Moreover, the prevalence of severe neonatal asphyxia was higher in sIUGR pregnancies with PE than in normal-growth pregnancies with PE (8.8% versus 2.5%, p = 0.020). CONCLUSIONS: sIUGR is associated with increased odds of developing severe PE in twin pregnancies, leading to poorer perinatal outcomes.