Discovery of Highly Selective, Potent, Covalent, and Orally Bioavailable Factor XIIa Inhibitors for the Treatment of Thrombo-Inflammation.

Thrombo-inflammation is closely associated with a few severe cardiovascular and infectious diseases. Factor XIIa (FXIIa) in the intrinsic coagulation pathway plays a pivotal role in the development of thrombo-inflammation and its inhibition has emerged as a potential therapeutic approach for thrombo-inflammatory disorders. Nonetheless, as of now, few small-molecule FXIIa inhibitors have demonstrated notable effectiveness against thrombo-inflammation, with none progressing into clinical stages. Herein, we present potent, covalent, reversible, and selective small-molecule FXIIa inhibitors such as 4a and 4j obtained through structure-based drug design. Compounds 4a and 4j showed significant anticoagulation and substantial anti-inflammatory effects in vitro, coupled with exceptional plasma stability. Furthermore, in carrageenan-induced thrombosis models, 4a and 4j demonstrated remarkable dual antithrombotic and anti-inflammatory activity when administered orally. Compound 4j exhibited a favorable safety profile without obvious tissue toxicity in mice, suggesting its potential as an oral therapeutic option for thrombo-inflammation.

Prevalence and related factors of children myopia in Pudong New Area, Shanghai: a cross-sectional study.

OBJECTIVES: This study aimed to assess the prevalence and related factors of myopia among school-aged children after COVID-19 pandemic. DESIGN: Cross-sectional study. SETTING: Pudong New Area, Shanghai. PARTICIPANTS: 1722 children aged 7-9 randomly selected from 8 primary schools were screened from 1 February 2023 to 30 April 2023. MAIN OUTCOME MEASURES: Children's height, weight and eye parameters were examined. Myopia was defined as a cycloplegic spherical equivalent ≤-0.50 dioptres in either eye. A vision-related behaviour questionnaire was applied to investigate the associations between myopia and its risk factors. RESULTS: Of the 1722 individuals enrolled, 25.6% (456) had myopia. After adjusting other characteristics, the following factors were associated with an increased rate of myopia: age (9 years vs 7 years, adjusted OR (AOR) 1.84, 95% CI 1.18 to 2.85, p=0.007), parental myopia status (both myopia vs none, AOR 5.66, 95% CI 3.71 to 8.63, p<0.001; one myopia vs none, AOR 2.92, 95% CI 1.93 to 4.42, p<0.001), reading books too close (yes vs no, AOR 1.58, 95% CI 1.20 to 2.08, p=0.001), writing with a tilted head (yes vs no, AOR 1.37, 95% CI 1.05 to 1.77, p=0.019), sleep patterns (early to bed late to rise vs early to bed early to rise, AOR 1.52, 95% CI 1.02 to 2.26, p=0.039). By contrast, a higher monthly household income and the habit of reading while lying down were associated with lower risk of myopia. CONCLUSIONS: The prevalence of myopia is of concern among young school-aged children after COVID-19. Correcting eye use behaviour and improving sleep habits may reduce myopia. Also, gender differences should be considered in prevention strategies for children's myopia.

Placental homing peptide guides HIF1α­silenced exosomes conjugates for targeted enhancement of invasion of trophoblast cells.

There is a current lack of availability of therapeutics to treat Preeclampsia (PE), primarily due to the risk of harm to the fetus. Hypoxia­inducible factor­1α (HIF1α) is highly expressed in trophoblast cells and suppresses their invasive ability. Extensive studies have confirmed the positive effects of mesenchymal stem cell (MSC)­derived exosomes on PE. The aim of the present study was to develop a method for targeted delivery of HIF1α­silenced exosomes to the placenta. HIF1α was overexpressed in JEG­3 cells. Then, the glucose uptake, lactate production, proliferation and invasion of HIF1α­elevated JEG­3 cells were detected. Exosomal membrane protein lysosome­associated membrane glycoprotein 2b and placental homing peptide CCGKRK gene sequence amplified by PCR were conjugated using short hairpin RNA­HIF1α (sh­HIF1α) sequence (exo­pep­sh­HIF1α), which were then transfected into MSCs cultured in vitro. Exosomes were isolated from the supernatant of the aforementioned MSCs and identified by determining the size and exosomal markers. Finally, the invasion ability of MSCs­derived exosomes treated JEG­3 cells were detected using Transwell assays. HIF1α was demonstrated to remarkably promote the uptake of glucose and the production of lactate in JEG­3 cells. In addition, high levels of HIF1α facilitated the proliferation of JEG­3 cells, while suppressing their invasion ability. Bone marrow derived MSCs were cultured in vitro and exosomes were successfully isolated from these cells. Exo­pep­sh­HIF1α significantly reduced placental HIF1α expression, and induced significant enhancement of placental invasion. Overall, placental homing peptide­guided HIF1α­silenced exosomes effectively facilitated the invasion of placental trophoblasts, which could be used for the targeted delivery of payloads to the placenta and serve as a novel placenta­specific therapeutic approach.

Risk factors and prognostic factors for pulmonary large cell neuroendocrine carcinoma with brain metastasis.

BACKGROUND: As the studies regarding the brain metastasis (BM) of pulmonary large cell neuroendocrine carcinoma (LCNEC) are insufficient, the present research aims to describe the risk factors and prognostic factors that are related to cancer-specific survival (CSS) for LCNEC patients with BM. METHODS: The data of LCNEC patients between January 2010 and October 2018 were obtained from the SEER database. Binary logistic regression analyses were utilized to screen the possible risk factors related to BM. Prognostic factors for LCNEC patients with BM were indentified by Cox regression analyses. Moreover, a nomogram was established to predict the 6-, 12-, and 18-month CSS rates. The concordance index (C-index), receiver operating characteristic (ROC) curves and calibration curves were utilized to assess the discrimination and reliability of the model. Clinical decision curves (DCAs) were used to evaluate the clinical benefits and utility of our model. RESULTS: Totally, 1875 patients were enrolled, with 294 (15.7%) of them having BM at diagnosis. Multivariate logistic regression analyses revealed that patients with age < 65 (odds ratio, OR = 1.564) and N2 staging (OR = 1.775) had a greater chance of developing BM. Age (≥ 65 vs. < 65: hazard ratio, HR = 1.409), T staging (T1 vs. T0: HR = 4.580; T2 vs. T0: HR = 6.008; T3 vs. T0: HR = 7.065; T4 vs. T0: HR = 6.821), N staging (N2 vs. N0: HR = 1.592; N3 vs. N0: HR = 1.654), liver metastasis (HR = 1.410), primary site surgery (HR = 0.581) and chemotherapy (HR = 0.452) were independent prognostic factors for LCNEC patients with BM. A nomogram prediction model was constructed by incorporating these factors. Using the C-index, calibration curves, ROC curves, and DCAs, we found that the clinical prediction model performed well. CONCLUSION: We described the risk factors and prognostic factors that were associated with CSS for LCNEC patients with BM. The related nomogram was established and validated to help clinicians formulate more rational and effective treatment strategies.

Short-Term effects of ambient ozone on the risk of conjunctivitis outpatient visits: a time-series analysis in Pudong New Area, Shanghai.

To date, the relationship between conjunctivitis and air pollutants has been widely concerned, but the conclusions are not very unified. This study aims to explore the short-term effects of ambient ozone (O3) on the conjunctivitis outpatient visits in Pudong New Area, Shanghai. A quasi-Poisson model combined with the distributed lag nonlinear model (DLNM) was performed to study the short-term effects of O3 on the risk of outpatient visits for conjunctivitis, after controlling the effects of temperature, relative humidity and wind speed. Taking the median concentration of O3 as a reference, the moderate high O3 (75th percentile) showed the largest effect estimates for single and cumulative lag effects at lag 4 (RR 1.013, 95% confidence interval [CI] 1.006-1.019) and lag 0-10 (RR 1.075, 95%CI 1.025-1.128), respectively. Our study suggested that the moderate high O3 increased the chances of conjunctivitis outpatient visits and had an evident lag effect.

Mesenchymal Stem Cell-Derived Exosomal Long Noncoding RNA MALAT1-201 Regulated the Proliferation, Apoptosis and Migration of Trophoblast Cells via Targeting miR-141.

OBJECTIVE: MALAT1 has been confirmed to play a vital role in the progression of preeclampsia (PE). However, as one of the spliceosomes of MALAT1, the role and mechanism of MALAT1-201 in the progression of PE remain elusive. Mesenchymal stem cells (MSCs) correlate with angiogenesis and trophoblast formation and could maintain successful pregnancy, while the molecular mechanisms are still unclear. The aim of the study was to investigate the role and potential mechanism of MALAT1-201 in PE. METHODS: We isolated MSCs from bone marrow and cultured in vitro. We overexpressed MALAT1-201 in MSCs and collected exosomes released by MSCs to treat trophoblast cells. Then, the proliferation, apoptosis and migration of MALAT1-201 elevated trophoblast cells were detected by CCK-8, flow cytometer and transwell assay, respectively. The binding site between MALAT1-201 and miR-141 was detected by dual-luciferase assays. The location of MALAT1-201 was detected by cytoplasmic and nuclear RNA fractionation. RESULTS: We successfully cultured bone marrow derived MSCs in vitro. MSC-Ex carrying MALAT1-201 promoted proliferation and migration, while suppressed apoptosis of trophoblast cells, which is similar to the effects of MALAT1-201 gene on trophoblast cells. In addition, MALAT1-201 was mainly localized in the nucleus and miR-141 was the target of MALAT1-201. CONCLUSIONS: MALAT1-201 derived from MSC-Ex regulated the proliferation, apoptosis and migration of HTR8-Svneo cells by targeting miR-141, which indicated a promising therapeutic target for PE.

Hes-related family BHLH transcription factor with YRPW motif 1-activated proteasome 26S subunit, non-ATPase 14 regulates trophoblast function and endometrial angiogenesis.

Proteasome 26S subunit, non-ATPase 14 (PSMD14) expression has been previously reported to be reduced in patients with pre-eclampsia (PE). The present study investigated the interaction network associated with the role of PSMD14 in PE. Reverse transcription-quantitative PCR (RT-qPCR) and western blotting were performed to determine the transfection efficacy following plasmid-based gene transfer of PSMD14 into HTR-8/SVneo cells. Cell proliferation was measured using an MTT assay and 5-ethynyl-2'-deoxyuridine staining. The expression of proliferation-related proteins, including Ki67 and PCNA, was determined using western blotting. Wound healing and Transwell assays were performed to measure cell invasion and migration, whilst the expression of migration-related proteins, including MMP2 and MMP9, was measured using western blotting. The angiogenesis of HUVECs following treatment with the HTR-8/SVneo cell culture supernatant was examined using tube formation assay. Following overexpression of Hes-related family BHLH transcription factor with YRPW motif 1 (HEY1) by transfection of pcDNA3.1 expression vector containing full-length human HEY1 or knockdown by transfection of shRNA plasmids targeting HEY1, the expression of HEY1 and PSMD14 was detected using RT-qPCR and western blotting. The potential interaction between HEY1 and the PSMD14 promoter was examined using dual-luciferase reporter and chromatin immunoprecipitation assays. PSMD14 overexpression was found to promote the proliferation, invasion, migration of HTR-8/SVneo cells and the angiogenesis of HUVECs following treatment with the HTR-8/SVneo cell culture supernatant, accompanied by enhanced expression of proliferation and migration-related proteins. Furthermore, the transcription factor HEY1 activated the expression of PSMD14. Knocking down HEY1 expression partially reversed the promoting effects of PSMD14 overexpression on the proliferation, invasion, migration, angiogenesis, proliferation and migration-related protein expression in trophoblasts. In conclusion, HEY1-activated PSMD14 promoted trophoblast proliferation, invasion and angiogenesis. Therefore, HEY1 and PSMD14 can be potential targets for PE treatment.

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.

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.

Preparation, evaluation and application of molecularly imprinted solid-phase microextraction monolith for selective extraction of pirimicarb in tomato and pear.

A simple, rapid and sensitive method for the determination of pirimicarb in tomato and pear using polymer monolith microextraction (PMME) based on the molecularly imprinted polymer (MIP) monolith combined with high-performance liquid chromatography-photodiodes array detector (HPLC-PAD) was developed. By optimizing the polymerization conditions, such as the nature of porogenic solvent and functional monomer, the molar ratio of the monomer and cross-linker, an pirimicarb MIP monolith was synthesized in a micropipette tip using methacrylic acid (MAA) as the functional monomer, ethylene dimethacrylate (EGDMA) as the cross-linker and the mixture of toluene-dodecanol as the porogenic solvent. The MIP monolith showed highly specific recognition for the template pirimicarb. The monolith was applied for the selective extraction of pirimicarb in tomato and pear. Several parameters affecting MIP-PMME were investigated, including the nature and volume of extraction solvent, sample volume, flow rate and sample pH. Under the optimum PMME and HPLC conditions, the linear ranges were 2.0-1400 µg/kg for pirimicarb in tomato and pear with the correlation coefficient of above 0.999. The detection limits (s/n=3) were both 0.6 µg/kg. The proposed method was successfully applied for the selective extraction and determination of pirimicarb in tomato and pear.