Alterations of plasma circulating microRNAs in BALB/c mice with Toxocara canis visceral and cerebral larva migrans.

BACKGROUND: Human toxocariasis is a neglected parasitic disease characterised by the syndromes visceral, cerebral, and ocular larva migrans. This disease is caused by the migrating larvae of Toxocara roundworms from dogs and cats, affecting 1.4 billion people globally. Via extracellular vesicles (EVs), microRNAs have been demonstrated to play roles in host-parasite interactions and proposed as circulating biomarkers for the diagnosis and follow-up of parasitic diseases. METHODS: Small RNA-seq was conducted to identify miRNAs in the infective larvae of T. canis and plasma EV-containing preparations of infected BALB/c mice. Differential expression analysis and target prediction were performed to indicate miRNAs involved in host-parasite interactions and miRNAs associated with visceral and/or cerebral larva migrans in the infected mice. Quantitative real-time polymerase chain reaction (PCR) was used to amplify circulating miRNAs from the infected mice. RESULTS: This study reports host and parasite miRNAs in the plasma of BALB/c mice with visceral and cerebral larva migrans and demonstrates the alterations of these miRNAs during the migration of larvae from the livers through the lungs and to the brains of infected mice. After filtering unspecific changes in an irrelevant control, T. canis-derived miRNAs and T. canis infection-induced differential miRNAs are predicted to modulate genes consistently involved in mitogen-activated protein kinase (MAPK) signalling and pathways regulating axon guidance and pluripotency of stem in the infected mice with visceral and cerebral larva migrans. For these plasma circulating miRNAs predicted to be involved in host-parasite crosstalk, two murine miRNAs (miR-26b-5p and miR-122-5p) are experimentally verified to be responsive to larva migrans and represent circulating biomarker candidates for visceral and cerebral toxocariasis in BALB/c mice. CONCLUSIONS: Our findings provide novel insights into the crosstalk of T. canis and the mammalian host via plasma circulating miRNAs, and prime agents and indicators for visceral and cerebral larva migrans. A deep understanding of these aspects will underpin the diagnosis and control of toxocariasis in humans and animals.

Mode of delivery and neonatal outcomes in preterm twins less than 32 weeks of gestation or birthweight under 1500 g: a systematic review and meta-analyses.

BACKGROUND: The mode of delivery for twins born before 32 weeks of gestation remains controversial. Our purpose is to conduct a meta-analysis of twin pregnancies less than 32 weeks or twin weight less than 1500 g, so as to find a suitable delivery mode. METHODS: We searched PubMed database, Cochrane Library database, and EMBASE database through December 2022. This protocol was registered with PROSPERO (CRD42023386946) prior to initiation. Studies that compared vaginal delivery to cesarean section for newborns less than 32 weeks of gestation or birthweight under 1500 g were included. The primary result was neonatal mortality rate. Secondary result was neonatal morbidity. The quality of literatures included in the research was evaluated in accordance with Newcastle-Ottawa Scale (NOS) literature quality evaluation scale. We use odds ratio (OR) as the effect index for binary variables. Point estimates and 95% confidence intervals (95% CI) were calculated. P < 0. 05 indicated statistically significant difference. RESULTS: Our search generated 5310 articles, and a total of 8 articles comprising a total of 14,703 newborns were included in the analysis. The odds ratios of neonatal mortality rate were for twins delivered by vaginal delivery compared to cesarean section were 0.84 (95% CI 0.57-1.24, P = 0.38). The 5-min Apgar score < 7 (95% CI 0.44-1.75, P = 0.72), necrotizing enterocolitis (95% CI 0.81-1.19, P = 0.82), intraventricular hemorrhage (95% CI 0.41-1.86, P = 0.71), periventricular leukomalacia (95% CI 0.16-4.52, P = 0.84), bronchopulmonary dysplasia (95% CI 0.88-1.36, P = 0.42), and respiratory distress syndrome (95% CI 0.23-2.01, P = 0.48) were not statistically significant between the two groups. CONCLUSION: We have observed that vaginal delivery does not confer an increased risk of neonatal morbidity and mortality in twins born before 32 weeks of gestation. However, the current results are affected by substantial heterogeneity and confounding factors. We still need high-quality randomized-controlled studies require to address this important question.

MiRNA-26a-5p inhibits preterm labor initiation by targeting and regulating TRPC3 ion channel protein expression.

The incidence of preterm birth (PTB) is increasing annually worldwide, leading to various health problems or even fetal deaths. Our previous work demonstrated the activation of transient receptor potential cation channel subfamily C 3 (TRPC3) in mice with PTB, and its activation could promote inward flow of calcium ions and uterine smooth muscle (USM) contraction via regulation of Cav3.2, Cav3.1, and Cav1.2. However, the upstream regulators of TRPC3 and its mechanisms remain unknown. In the present study, the binding of miR-26a-5p to the 3' untranslated region of TRPC3 was predicted by bioinformatics databases (TargetScanHuman and starBase v3.0) and confirmed by a dual-luciferase assay. MiR-26a-5p was downregulated, while TRPC3 was upregulated in the USM tissues of patients with PTB compared to people without PTB. The results showed that miR-26a-5p mimic transfection markedly reduced TRPC3 expression in LPS-stimulated USM cells. Additionally, miR-26a-5p regulated intracellular Ca2+ levels in USM cells by targeting TRPC3. Furthermore, miR-26a-5p inhibited the CPI17/PKC/PLCγ signaling pathway and reduced the expression of Cav3.2, Cav3.1, and Cav1.2. In conclusion, miR-26a-5p regulated the initiation of PTB by targeting TRPC3 and regulating intracellular Ca2+ levels. This study provides a promising diagnostic biomarker and therapeutic target for PTB.

The Safety and Antiaging Effects of Nicotinamide Mononucleotide in Human Clinical Trials: an Update.

The importance of nicotinamide adenine dinucleotide (NAD+) in human physiology is well recognized. As the NAD+ concentration in human skin, blood, liver, muscle, and brain are thought to decrease with age, finding ways to increase NAD+ status could possibly influence the aging process and associated metabolic sequelae. Nicotinamide mononucleotide (NMN) is a precursor for NAD+ biosynthesis, and in vitro/in vivo studies have demonstrated that NMN supplementation increases NAD+ concentration and could mitigate aging-related disorders such as oxidative stress, DNA damage, neurodegeneration, and inflammatory responses. The promotion of NMN as an antiaging health supplement has gained popularity due to such findings; however, since most studies evaluating the effects of NMN have been conducted in cell or animal models, a concern remains regarding the safety and physiological effects of NMN supplementation in the human population. Nonetheless, a dozen human clinical trials with NMN supplementation are currently underway. This review summarizes the current progress of these trials and NMN/NAD+ biology to clarify the potential effects of NMN supplementation and to shed light on future study directions.

Recent Advances and Future Prospects of Mycosporine-like Amino Acids.

Mycosporine-like amino acids (MAAs) are a class of water-soluble active substances produced by various aquatic organisms. However, due to the limitations of low accumulation of MAAs in organisms, the cumbersome extraction process, difficult identification, and high cost, MAAs have not yet been widely used in human life. Recently, there has been an emergence of heterologous synthesis for MAAs, making increasing yield the key to the quantification and application of MAAs. This review summarizes the latest research progress of MAAs, including: (1) introducing the biodistribution of MAAs and the content differences among different species to provide a reference for the selection of research subjects; (2) elaborating the species and molecular information of MAAs; (3) dissecting the synthesis mechanism and sorting out the synthesis pathways of various MAAs; (4) summarizing the methods of extraction and identification, summarizing the advantages and disadvantages, and providing a reference for the optimization of extraction protocols; (5) examining the heterologous synthesis method; and (6) summarizing the physiological functions of MAAs. This paper comprehensively updates the latest research status of MAAs and the various problems that need to be addressed, especially emphasizing the potential advantages of heterologous synthesis in the future production of MAAs.

The overgeneralization of pain-related fear in individuals with higher pain sensitivity: A behavioral and event-related potential study.

Fear generalization contributes to the development and maintenance of pain. Pain sensitivity has been proposed to predict the strength of fear responses to aversive stimuli. However, whether individual variation in pain sensitivity affects pain-related fear generalization and its underlying cognitive processing remains unclear. To address this gap, we recorded behavioral and event-related potential (ERP) data among 22 high pain sensitivity (HPS) and 22 low pain sensitivity (LPS) healthy adults when exposed to a fear generalization paradigm. The behavioral results indicate that the HPS group displayed higher unconditioned stimulus expectancy and greater fear, arousal, and anxiety ratings to conditioned stimulus and generalization stimulus than the LPS group (all p values < 0.05). The ERP results showed that the HPS group exhibited a larger late positive potential evoked by GS2, GS3 and CS- (all p < 0.005) but a smaller N1 evoked by all CS and GSs (all p values < 0.05) relative to the LPS group. These findings suggest that individuals with a high level of pain sensitivity allocate more attention resources to pain-related threatening stimuli, which contributes to an overgeneralization of pain-related fear.

The excessive generalization of fear affected by perceptual bias in experimental pain individuals: Evidence from an event-related potential study.

INTRODUCTION: Excessiv generalization of fear contributes to the development and maintenance of pain. Prior research has demonstrated the importance of perception in fear generalization and found that individuals in painful conditions exhibited perceptual bias. However, the extent to which perceptual bias in pain affects the generalization of pain-related fear and its underlying neural activity remains unclear. METHODS: Here, we tested whether perceptual bias in experimental pain individuals led to the overgeneralization of pain-related fear by recording behavioral and neural responses. To this end, we established an experimental pain model by spraying capsaicin on the surface of the seventh cervical vertebra of the participant. A total of 23 experimental pain participants and 23 matched nonpain controls learned fear conditioning and then completed the fear generalization paradigm combined with the perceptual categorization task. RESULTS: We found that the novel and safety cues were more likely to be identified as threat cues in the experimental group, resulting in higher US expectancy ratings compared to the control group. The event-related potential results showed that the experimental group exhibited earlier N1 latency and smaller P1 and late positive potential amplitudes than those in the control group. CONCLUSION: Our findings suggest that the experimental pain individuals exhibited an excessive generalization of fear affected by perceptual bias and reduced their attentional allocation to pain-related fear stimuli.

AQP9 (Aquaporin 9) Determines Arsenic Uptake and Tolerance in Human Hepatocellular Carcinoma Cells In Vitro.

Arsenic-based therapeutic strategies, even though promising for acute promyelocytic leukemia (APL), are limited by arsenic-related toxic effect and resistance with unknown mechanisms. The purpose of this study is to better understand the different sensitivities of hepatocellular carcinoma cells to arsenic and its mechanism. Arsenic-sensitive liver cancer cell line (HepG2) and arsenic-resistant HepG2 (AsHepG2) cells are employed to study the role of aquaporin 9 (AQP9) in arsenic uptake and tolerance. The half-maximal inhibitory concentration (IC50) value of arsenic in AsHepG2 cells (15.59 ± 1.36 µM) is significantly higher than that in HepG2 cells (7.33 ± 0.93 µM; p= 0.0288). We demonstrated that, with the treatment of sodium arsenite (NaAsO2), arsenic was accumulated at a significantly lower level in AsHepG2 cells in comparison with HepG2 cells (p= 0.00549). Further, arsenic level in AsHepG2 cells reaches a plateau after six hours of treatment, whereas arsenic continues to increase in HepG2 cells during the entire experimental period. Mechanistic study showed that the expression of AQP9 is decreased in a dose-dependent manner in AsHepG2 cells, but no significant difference in HepG2 cells. Furthermore, NaAsO2 dramatically increases AQP9 and p38 phosphorylation, which may partially regulate arsenic sensitivity in both cell lines. In conclusion, the expression and phosphorylation of AQP9 regulated by p38 kinase are involved in the arsenic uptake, thus regulating cellular arsenic sensitivity.

Effect of transcranial direct-current stimulation on executive function and resting EEG after stroke: A pilot randomized controlled study.

BACKGROUND: The effects of transcranial direct current stimulation (tDCS) on post-stroke executive impairment (PSEI) remain controversial. Resting stateelectroencephalogram (EEG) can assist in the diagnosis and assessment of executive dysfunction. OBJECTIVES: We aimed to use EEG to explore the effect of tDCS on executive function among stroke patients. METHODS: Twenty-four patients with PSEI were randomly divided into experimental and control groups, which received real and sham stimulation, respectively. Anodal electrical stimulation was applied to the left dorsolateral prefrontal lobe (F3). The stimulation intensity was 2 mA for 20 min once daily for 7 days. Executive function was monitored using neuropsychological scales. RESULTS: The experimental group outperformed the control group in clinical scale results, with significant differences in the following scores: symbol digital modalities test, TMT-A, TMT-B, and digital span test. In the left central zone, theta band power was significantly higher after anodal electrical stimulation than before. Analysis of the correlation between EEG power and psychometric scores revealed that the power change was positively correlated with the scores on the symbol digital modality test (r = 0.435, p < 0.05). CONCLUSION: Anodal tDCS can enhance executive function in patients with PSEI, and tDCS-related improvements are related to the enhancement of theta power in the affected region.

Visually Perceived Negative Emotion Enhances Mismatch Negativity but Fails to Compensate for Age-Related Impairments.

Objective: Automatic detection of auditory stimuli, represented by the mismatch negativity (MMN), facilitates rapid processing of salient stimuli in the environment. The amplitude of MMN declines with ageing. However, whether automatic detection of auditory stimuli is affected by visually perceived negative emotions with normal ageing remains unclear. We aimed to evaluate how fearful facial expressions affect the MMN amplitude under ageing. Methods: We used a modified oddball paradigm to analyze the amplitude of N100 (N1) and MMN in 22 young adults and 21 middle-aged adults. Results: We found that the amplitude of N1 elicited by standard tones was smaller under fearful facial expressions than neutral facial expressions and was more negative for young adults than middle-aged adults. The MMN amplitude under fearful facial expressions was greater than neutral facial expressions, but the amplitude in middle-aged adults was smaller than in young adults. Conclusion: Visually perceived negative emotion promotes the extraction of auditory features. Additionally, it enhances the effect of auditory change detection in middle-aged adults but fails to compensate for this decline with normal ageing. Significance: The study may help to understand how visually perceived emotion affects the early stage of auditory information processing from an event process perspective.

Effects of Misoprostol on Induction of Labour in Patients with Hypertensive Disorders of Pregnancy: A Meta-Analysis.

Objective: Hypertensive disorders of pregnancy (HDP) can cause serious prenatal and postnatal complications and is a threat to maternal and fetal health. To offer guidance for clinical decisions, we systematically reviewed the effects of misoprostol on induction of labour in HDP patients. Methods: PubMed, Web of Science, Embase, CNKI, and Wanfang databases were searched for relevant literature from 2010 to 2020. Subsequently, a meta-analysis was performed to compare the effective rate of induction of labour and reducing postpartum hemorrhage (PPH) between the intervention group (n = 544, misoprostol) and the control group (n = 543, oxytocin). Results: A total of 10 studies with 1087 patients were included. The 10 studies compared the effective rate of induction of labour between the two groups and confirmed that the effective rate in the intervention group was significantly higher than that in the control group (OR = 4.37; 95% CI: 2.73, 7.00). Seven studies compared PPH between the groups and showed that it was significantly reduced in the intervention group compared to the control group (SMD = -1.32; 95% CI: -2.05, -0.59; P < 0.0001). Conclusion: Misoprostol has a high effective rate of induction of labour in HDP patients and is an effective uterotonic agent in reducing PPH. This meta-analysis provides clinicians with meaningful information to help them make evidence-based decisions.

The novel hybrid polycarbonate polyurethane / polyester three-layered large-diameter artificial blood vessel.

BACKGROUND: The most common materials of artificial blood vessels are polyethylene terephthalate and polytetrafluoroethylene. But polycarbonate polyurethane (PCU) is an ideal material for vascular prostheses because of their excellent characteristics. As far as we know, our artificial blood vessel is the first type of hybrid PCU/polyester three-layered large-diameter artificial blood vessel in the world. OBJECTIVE: The purpose of this preclinical animal experiment is to evaluate the hemocompatibility, histocompatibility, effectiveness, and safety of the three-layered large-diameter artificial blood vessel in sheep. METHODS: The artificial blood vessels took place of the initial segments of the sheep's thoracic aorta by end-to-end anastomosis. RESULTS: All of the 14 sheep are male, their average body weight (BW) was 30.57 ± 3.95 kg. All 14 artificial blood vessels successfully replaced the thoracic aortas. 5 sheep did not survive to the end of the experiment, while the remaining 9 sheep did. After the surgery, the blood biochemical and blood routine indicators fluctuate slightly within the normal range. The angiography showed that the implanted artificial blood vessels were unobstructed without obvious stenosis or expansion. 24 weeks after surgery, the lumen surfaces of the artificial blood vessels were covered by endothelia in different degrees, and the average endothelialization rate was 69.44% (range: 20% to 100%). CONCLUSIONS: This artificial blood vessel is the first to use PCU in large-diameter artificial vascular grafts. It has excellent blood compatibility, wonderful biocompatibility, high endothelialization rate, and 100% patency.

Effect of Low-Frequency Repetitive Transcranial Magnetic Stimulation on Executive Function and Its Neural Mechanism: An Event-Related Potential Study.

Objective: Executive function refers to the conscious control of thinking and behavior in psychological process. Executive dysfunction widely exists in a variety of neuropsychiatric diseases, and is closely related to the decline of daily living ability and function. This study intends to explore the effect of low-frequency repetitive transcranial magnetic stimulation (rTMS) on executive function and its neural mechanism by using event-related potential (ERP), so as to provide basis for further study on the relationship between cerebral cortex and executive function. Methods: Task switching paradigm was used to study the cognitive flexibility in executive function. Thirty-one healthy subjects were randomly assigned to receive rTMS stimulations (1 Hz rTMS or sham rTMS) to the left dorsolateral prefrontal cortex (DLPFC) twice. The switching task and the electroencephalography EEG recordings were performed before (pre-rTMS/pre-sham rTMS) and immediately after the end of the rTMS application (post-rTMS/post-sham rTMS). Results: The analysis of RTs showed that the main effects of switching and time were statistically significant. Further analysis revealed that the RT of rTMS stimulation was longer than sham rTMS at post-stimulation. ERP analysis showed that there was a significant switching effect in frontal and central scalp location, and the P2 amplitude in switch trials was greater than that in non-switch trials. At post-stimulation, the N2 amplitude of rTMS is more negative than that of sham rTMS at non-switch trials, whereas no such difference was found at switch trials. The P3 amplitude and LPC amplitude are significantly reduced by rTMS at post-stimulation. Conclusion: Low-frequency rTMS of the left DLPFC can cause decline of cognitive flexibility in executive function, resulting in the change of N2 amplitude and the decrease of P3 and LPC components during task switching, which is of positive significance for the evaluation and treatment of executive function.

The Inhibitory Effect of Emotional Conflict Control on Memory Retrieval.

Evidence is mounting that emotional conflict is mainly resolved by the rostral anterior cingulate inhibiting the processing of emotional distractors. However, this theory has not been verified from the perspective of memory retrieval. This experiment aimed to explore the offline effect of emotional conflict processing on memory retrieval. We adopted a modified encoding-retrieval paradigm to explore this issue. Participants' electroencephalography (EEG) signal were also collected. A face-word Stroop task was used to create the congruency factor. In addition, an old/new judgment task was used to evaluate the recognition performance. During the retrieval phase, the response time of the incongruent condition was longer and the recognition accuracy was lower compared with congruent and neutral conditions in the behavioral data. For event-related potentials (ERP), we detected two well-established old/new effects related to memory retrieval under both neutral and emotional conditions: the frontal negativity (FN400) related to familiarity-driven recognition and the late posterior negativity (LPN) related to reconstructive processing or evaluation of retrieval outcomes. More importantly, the old/new effects were missing for incongruent condition during the early stage of FN400 (300-400 ms). Besides, for LPN (700-900 ms), the old/new effects of the incongruent condition are greater than the congruent condition. The results prove that the encoding phase's emotional congruency factor has a regulatory effect on the retrieval phase's early familiarity processing and evaluation of retrieval outcomes. Our data confirm the inhibitory effect of emotional conflict control on memory retrieval and support the emotional conflict control mechanism found in previous studies.

Removal of Sb(III) by sulfidated nanoscale zerovalent iron: The mechanism and impact of environmental conditions.

Pollution of Sb(III) in water has caused great concern in recent years. Nanoscale zero-valent iron (nZVI) can detoxify Sb(III) polluted water, but the rapid passivation and low adsorption capacity limit its practical application. Hence, this study provides a new and efficient nanotechnology to remove Sb(III) using the sulfidated nanoscale zero-valent iron (S-nZVI). The S-nZVI exhibits higher Sb(III)-removal efficiency than pristine nZVI under both aerobic and anoxic conditions. The adsorption capacity of Sb(III) by optimized S-nZVI (465.1 mg/g) is 6 times as high as that of the pristine nZVI (83.3 mg/g) under aerobic conditions. The results indicate that Sb(III) and Sb(V) can be immobilized on the surface of S-nZVI by forming Fe-S-Sb precipitates. Moreover, characterization results demonstrate that the existence of S2- can not only activate H2O2 to produce hydroxyl radical, but also accelerate the cycle of Fe3+/Fe2+ to improve the efficiency of Fenton reaction. Therefore, S-nZVI can produce more hydroxyl radicals to oxidize Sb (III) to Sb (V) and results in 2.3-fold higher oxidation rate of Sb(III) compared to pristine nZVI. The formed FeS layer on the S-nZVI surface can also improve the release ability of Fe2+ and accelerate the formation of nZVI corrosion products. S-nZVI thus holds great potential to be applied in antimony removal.

[Corrigendum] Roles and mechanisms of TRPC3 and the PLCγ/PKC/CPI-17 signaling pathway in regulating parturition.

Subsequent to the publication of the above paper, the authors have realized that there were errors in Fig. 1 on page 902; essentially, the experimental data described as being 'Preterm' and 'Infected preterm' should have been labelled as 'Full­term without labor' and 'Preterm', respectively. Note that these errors in Fig. 1 were not reflected in the published figure legend. The corrected version of Fig. 1 is shown here. The authors sincerely apologize for this mistake, and regret any inconvenience this mistake has caused. [the original article was published in the Molecular Medicine Reports 17: 898­910, 2018; DOI: 10.3892/mmr.2017.7998].

Roles and mechanisms of TRPC3 and the PLCγ/PKC/CPI-17 signaling pathway in regulating parturition.

The aim of the current study was to investigate the role of phospholipase C (PLC)γ/protein kinase C (PKC)/C­kinase­activated protein phosphatase­1 (CPI­17) signaling pathways in uterine smooth muscle during parturition. Samples of uterine tissue were collected from pregnant patients who underwent a caesarean section for preterm delivery, full­term delivery with labor onset, full­term delivery without labor onset, and from a non­pregnant control group undergoing surgery for cervical intraepithelial neoplasia III. Immunohistochemistry, and western blotting were used to assess the association between TRPC3 levels and parturition and the influence of calcium ion channels. In addition, pregnant mice were used to explore the effect of uterine canonical transient receptor potential 3 (TRPC3) expression on the parturition­triggering mechanism and PLCγ/PKC/CPI­17 signaling pathways. Pregnant mouse uterine smooth muscle cells were cultivated, with and without TRPC3 silencing, and the expression levels of PLCγ, PKC and CPI­17, the upstream and downstream factors of the TRPC3 pathway, were measured in pregnant mouse uterine smooth muscle cells, in order to provide a theoretical basis for the prevention and treatment of premature labor. In the preterm and full­term without labor onset patient groups, the TRPC3 gene expression in the mSMCs was significantly overexpressed when compared with the non­pregnant group (P<0.05); however, TRPC3 expression was not elevated in the full­term with labor onset group, exhibiting no significant difference compared with the non­pregnant group (P>0.05). During pregnancy, compared with the non­pregnant controls, Cav1.2, Cav3.1 and Cav3.2 gene expression levels were markedly increased (P<0.05) in mSMCs from the preterm delivery group and the full­term with labor onset group, however were non­significantly increased in the full­term without labor onset group. The level of TRPC3 was highest in the preterm group, while the levels of Cav1.2, Cav3.1 and Cav3.2 were highest in the full­term with labor onset group. In the preterm, LPS­treated preterm and full­term groups, TRPC3, MAPK, ERK1/2, P­ERK, Cav3.2, Cav3.1 and Cav1.2 were all expressed at higher levels than in the unfertilized group. In the LPS­treated preterm group, the levels of TRPC3, MAPK, ERK1/2, P­ERK, Cav3.2, Cav3.1 and Cav1.2 were increased compared with the preterm group. Furthermore, following transfection of small interfering TRPC3 (siTRPC3) into cells, it was demonstrated that the levels of TRPC3, PLCγ, PKC, CPI­17, P­CPI­17, Cav1.2, Cav3.1 and Cav3.2 expression were lower in the LPS siTRPC3 group when compared with that of the LPS­treated untransfected control group.

Enhanced expression of transient receptor potential channel 3 in uterine smooth muscle tissues of lipopolysaccharide-induced preterm delivery mice.

OBJECTIVES: We aimed to investigate the influence of transient receptor potential channel 3 (TRPC3) on lipopolysaccharide-induced (LPS) preterm delivery mice. MATERIALS AND METHODS: Mice were randomly assigned to the four groups: an unpregnant group, a mid-pregnancy group (E15), a term delivery group, and an LPS-induced preterm delivery group (intraperitoneal injection LPS at 15 days). Uterine smooth muscles were obtained through caesarean section; TRPC3 expression was measured by real-time PCR, western blotting, and immunohistochemistry. A specific inhibitor of TRPC3 (SKF96365) was injected into the LPS-induced preterm delivery group to determine whether the delivery interval was prolonged. RESULTS: TRPC3 was primarily expressed in the uterine smooth muscle layer. In addition, the LPS-induced preterm delivery group had an obviously higher expression level of TRPC3 mRNA and protein compared with the unpregnant and E15 groups, which were close to term delivery. More importantly, SKF96365 prolongs the delivery interval of LPS-induced preterm delivery mice. CONCLUSION: Enhanced expression of TRPC3 may be associated with LPS-induced preterm delivery in mice. The specific inhibitor of TRPC3 (SKF96365) may be helpful for clinical treatment of preterm delivery.

Lipopolysaccharides upregulate calcium concentration in mouse uterine smooth muscle cells through the T-type calcium channels.

Infection is a significant cause of preterm birth. Abnormal changes in intracellular calcium signals are the ultimate triggers of early uterine contractions that result in preterm birth. T­type calcium channels play an important role in the pathogenesis of cancer, as well as endocrine and cardiovascular diseases. However, there are limited studies on their role in uterine contractions and parturition. In the present study, mouse uterine smooth muscle cells were isolated and treated with lipopolysaccharides (LPS) to mimic the microenvironment of uterine infection in vitro to investigate the role of T­type calcium channels in the process of infection­induced preterm birth. The results from quantitative polymerase chain reaction and western blot analysis showed that LPS significantly induced the expression of the Cav3.1 and Cav3.2 subtypes of T­type calcium channels. Measurements of intracellular calcium concentration showed a significant increase in response to LPS. However, these effects can be reversed by T­type calcium channel blockers. Western blot analysis further indicated that LPS induced the activation of the nuclear factor (NF)­κB signaling pathway, and endothelin­1 (ET­1) was significantly upregulated, whereas NF­κB inhibitors significantly inhibited the LPS­induced upregulation of Cav3.1, Cav3.2 and ET­1 expression. In addition, ET­1 directly induced Cav3.1 and Cav3.2 expression, whereas ET­1 antagonists inhibited the LPS­induced upregulation of Cav3.1 and Cav3.2 expression. In conclusion, the present study demonstrates that infection triggers the upregulation of T­type calcium channels and promotes calcium influx. This process relies on the activation of the NF­κB/ET­1 signaling pathway. The T­type calcium channel is expected to become an effective target for the prevention of infection­induced preterm birth.

Progesterone receptor B promoter hypermethylation in human placenta after labor onset.

To determine the methylation status of progesterone receptor B (PR-B) promoter and how PR-B regulates progesterone action in placenta during human pregnancy. Placentas were obtained from the pregnancy women at term who underwent cesarean delivery and vaginal delivery. The methylation status of the PR-B promoter was analyzed using the methylation-specific polymerase chain reaction (PCR) and bisulfite sequencing PCR. And the messenger RNA (mRNA) and protein expression of the PR-B and DNA methyltransferases (DNMTs) were determined by quantitative real-time PCR and Western blot. Compared with the cesarean group, the placentas of vaginal delivery group had greater levels of PR-B DNA methylation, and the PR-B, DNMT1, DNMT3a, and DNMT3b mRNA and protein expression were significantly decreased. Progesterone receptor B methylation occurs with high frequency after labor onset and may play an important epigenetic mechanism of labor-related PR-B negative expression, thereby mediating the biological process of functional progesterone withdrawal at term for parturition.