Meiosis-mediated reproductive toxicity by fenitrothion in Caenorhabditis elegans from metabolomic perspective.

Fenitrothion (FNT), an organophosphorus insecticide, is widely detected in the living environment. The reproductive and endocrine toxicity of FNT to biological communities has been ever reported, but potential mechanism and reproductive toxicity dose effect remain unclear. In our study, we constructed Caenorhabditis elegans model to analyze the reproductive toxicity mechanism of FNT based on metabolomics and evaluated its reproductive toxicity dose effect using benchmark dose (BMD)method. Our results showed that FNT exposure significantly reduced brood size, number of germ cells, and delayed gonadal development in nematodes. Non-targeted metabolomics revealed that FNT exposure caused significant metabolic disturbances in nematodes, leading to a significant reduction in the synthesis of cortisol and melatonin, and the latter played a mediating role in the effects of FNT on number of germ cells. We further found that the levels of these two hormones were significantly negative correlated with the expression of the androgen receptor nhr-69 and affected the meiosis of germ cells by regulating the nhr-69/ fbf-1/2 /gld-3 /fog-1/3 pathway. Meanwhile, the study found the BMDL10s for N2 and him-5 mutant were 0.411 µg/L by number of germ cells and 0.396 µg/L by number of germ cells in the meiotic zone, respectively, providing a more protective reference dose for ecological risk assessment of FNT. This study suggested that FNT can affect androgen receptor expression by inhibiting cortisol and melatonin secretion, which further mediate the meiotic pathway to affect sperm formation and exert reproductive toxicity, and provides a basis for setting reproductive toxicity limits for FNT.

DNA methylation 6 mA and histone methylation involved in multi-/trans-generational reproductive effects in Caenorhabditis elegans induced by Atrazine.

Atrazine (ATR), a widely used triazine herbicide, is an environmental endocrine disruptor that can cause health problems. However, whether there are multi/trans-generational reproductive impacts of ATR have not been studied. Therefore, in this study, Caenorhabditis elegans was used as a preferable model organism to identify the multi/trans-generational reproductive toxicity of ATR. Only parental C.elegans (P0) were exposed to different concentrations (0.0004-40 mg/L) for 48 h and the subsequent offspring (F1-F5) were grown under ATR-free conditions and ATR conditions.The results showed that ATR exposure during P0 decreased fecundity, including a reduction in fertilized eggs, oocytes, and ovulation rate, delayed gonadal development, and decreased the relative area of gonad arm and germ cell number. Furthermore, continuous ATR exposure (P0-F5) causes a significant increase in reproductive toxicity in subsequent generations, although no significant toxicity occurred in the P0 generation after exposure to environmental-related concentrations, suggesting that ATR exposure might have cumulative effects. Likewise, parental exposure to ATR caused transgenerational toxicity impairments. Interestingly, only reproductive toxicity, not development toxicity, was transmitted to several generations (F1-F4), and the F2 generation showed the most notable changes. QRT-PCR results showed that genes expression related to DNA methylation 6 mA (damt-1, nmad-1) and histone H3 methylation (mes-4, met-2, set-25, set-2, and utx-1) can also be passed on to offspring. The function of H3K4 and H3K9 methylation were explored by using loss-of-function mutants for set-2, set-25, and met-2. Transmissible reproductive toxicity was absent in met-2(n4256), set-2(ok952), and set-25(n5021) mutants, which suggests that the histone methyltransferases H3K4 and H3K9 activity are indispensable for the transgenerational effect of ATR. Finally, the downstream genes of DNA methylation and histone H3 methylation were determined. ATR upregulated the expression of ZC317.7, hsp-6, and hsp-60. Mitochondrial stress in parental generation dependent transcription 6 mA modifiers may establish these epigenetic marks in progeny.

Female fecundability is associated with pre-pregnancy allostatic load: Analysis of a Chinese cohort.

INTRODUCTION: Allostatic load (AL) is a practical index that reflects multi-system physiological changes which occur in response to chronic psychosocial stress. This study investigated the association between female pre-pregnancy allostatic load and time to pregnancy. MATERIAL AND METHODS: We enrolled 444 women who met the inclusion criteria and were attempting to achieve pregnancy. Their allostatic load scores at baseline were evaluated by nine indicators (systolic blood pressure, diastolic blood pressure, fasting plasma glucose, plasma cortisol, noradrenaline, interleukin-6, hypersensitive C-reactive protein, high density lipoprotein cholesterol and body mass index). The participants were followed up and their pregnancy outcome ascertained 1 year later; we then calculated time-to-pregnancy. Cox models were used to estimate fecundability ratios and their 95% confidence intervals (95% CI) for different allostatic load scores. RESULTS: The median allostatic load score was 1 with a range of 0-6. The females were divided into four groups according to allostatic load score: group A (allostatic load = 0, 150/444, 33.8%), group B (allostatic load = 1-2, 156/444, 35.1%), group C (allostatic load = 3-4, 100/444, 22.5%) and group D (allostatic load = 5-6, 38/444, 8.6%). The cumulative pregnancy rate over 12 months for the four groups (A-D) was 55.4%, 44.5%, 50.9% and 26.9%, respectively (log-rank test, p = 0.042). After adjusting for potential confounding factors, group D showed a 59% reduction of fecundability compared with group A (fecundability ratio = 0.41, 95% CI 0.21-0.83). CONCLUSIONS: Women with a higher allostatic load score may have lower fecundability. Our findings suggest that the assessment of allostatic load during pre-conception consultation would be highly prudent.

The association between the pre-pregnancy vaginal microbiome and time-to-pregnancy: a Chinese pregnancy-planning cohort study.

BACKGROUND: Although sexually transmitted infections are regarded as the main cause of tubal infertility, the association between the common vaginal microbiome and female fecundability has yet to be determined. The objective of this study was to find convincing evidence relating to the impact of the vaginal bacterial structure on the fecundability of women planning pregnancy. METHODS: We recruited women who took part in the Free Pre-pregnancy Health Examination Project from 13 June 2018 to 31 October 2018 (n = 89, phase I) and from 1 November 2018 to 30 May 2020 (n = 389, phase II). We collected pre-pregnancy vaginal swabs from each subject; then, we followed up each subject to acquire the pregnancy-planning outcome in 1 year. In phase I, 16S rRNA gene sequencing was performed to investigate the vaginal bacterial content between the pregnancy and non-pregnancy groups. These findings were verified in phase II by applying a quantitative real-time polymerase chain reaction for the measurement of the absolute abundance of specific species. Cox models were used to estimate fecundability ratios (FR) for each vaginal microbiome type. RESULTS: In phase I, 59.6% (53/89) of women became pregnant within 1 year. The principal coordinate analysis showed that the pre-pregnancy vaginal microbial community structures of the pregnant and non-pregnant groups were significantly different (PERMANOVA test, R2 = 0.025, P = 0.049). The abundance of the genus Lactobacillus in the pregnancy group was higher than that of the non-pregnant group (linear discriminant analysis effect size (LDA) > 4.0). The abundance of the genus Gardnerella in the non-pregnant group was higher than those in the pregnant group (LDA > 4.0). In phase II, female fecundability increased with higher absolute loads of Lactobacillus gasseri (quartile Q4 vs Q1, FR = 1.71, 95%CI 1.02-2.87) but decreased with higher absolute loads of Fannyhessea vaginae (Q4 vs Q1, FR = 0.62, 95%CI 0.38-1.00). Clustering analysis showed that the vaginal microbiome of type D (characterized by a higher abundance of Lactobacillus iners, a lower abundance of Lactobacillus crispatus and Lactobacillus gassri) was associated with a 55% reduction of fecundability (FR = 0.45, 95%CI 0.26-0.76) compared with type A (featuring three Lactobacillus species, low Gardnerella vaginalis and Fannyhessea vaginae abundance). CONCLUSIONS: This cohort study demonstrated an association between the pre-pregnancy vaginal microbiome and female fecundability. A vaginal microbiome characterized by a higher abundance of L. iners and lower abundances of L. crispatus and L. gasseri appeared to be associated with a lower fecundability. Further research now needs to confirm whether manipulation of the vaginal microenvironment might improve human fecundability.

The current situation and future directions for the study on time-to-pregnancy: a scoping review.

INTRODUCTION: As problems associated with infertility and population aging increase, there is a growing interest in the factors that cause a decline in human fertility. Time-to-pregnancy (TTP) is a good indicator with which to reflect human fecundability. Here, we present a comprehensive overview of this topic. METHODS: Relevant qualitative and quantitative studies were identified by searching the Web of science and PubMed electronic databases. We included all literature, written in English, from inception to the 10th April 2021 providing the focus was on TTP. We conducted a narrative synthesis using thematic analysis. RESULTS: Traditional TTP-related study protocols include prospective and retrospective cohorts that provide a wealth of data to reveal potential influences on TTP. Thus far, a variety of factors have been shown to be associated with TTP in couples preparing for pregnancy, including basic demographic characteristics, menstrual status, chronic disease status, environmental endocrine disruptor exposure, and lifestyles. However, there are inevitable epidemiological bias in the existing studies, including recall bias, selection bias and measurement bias. Some methodological advances have brought new opportunities to TTP research, which make it possible to develop precision interventions for population fertility. Future TTP studies should take advantage of artificial intelligence, machine learning, and high-throughput sequencing technologies, and apply medical big data to fully consider and avoid possible bias in the design. CONCLUSION: There are many opportunities and future challenges for TTP related studies which would provide a scientific basis for the "precise health management" of the population preparing for pregnancy.

As the problems of infertility and population aging increase, there is a growing interest in the factors that cause a decline in human fertility. Time-to-pregnancy (TTP) is a good indicator with which to reflect human fecundability, and a longer TTP is known to reflect a reduction in fertility. Many original studies, with different designs, have used TTP to explore the factors that might influence fertility, including basic demographic characteristics, chronic disease status, environmental endocrine disruptor exposure, and lifestyles. However, much of the existing evidence is inconsistent and limited by various types of bias. This review provides a synopsis of recent TTP studies, and highlights new opportunities and future challenges.

The associations between low abundance of Mycoplasma hominis and female fecundability: a pregnancy-planning cohort study.

OBJECTIVE: To explore the impact of pre-pregnancy vaginal Mycoplasma hominis (M. hominis) colonization of low abundance on female fecundability. METHODS: In total, 89 females participating in a pre-pregnancy health examination program were included, and their pregnancy outcomes were followed up for 1 year. Vaginal swabs were collected, 16S rRNA genes were sequenced, and M. hominis colonization was confirmed by qPCR. Cox models were used to estimate the fecundability odds ratio (FOR) for women with M. hominis. RESULTS: The prevalence of M. hominis was 22.47% (20/89), and the abundance was relatively low (the cycle thresholds of the qPCR were all more than 25). In terms of the vaginal microbiome, the Simpson index of the positive group was significantly lower than that of the negative group (P = 0.003), which means that the microbiome diversity appeared to increase with M. hominis positivity. The relative abundance of M. hominis was negatively correlated with Lactobacillus crispatus (rho = - 0.24, P = 0.024), but positively correlated with Gardnerella vaginalis, Atopobium vaginae and Prevotella bivia (P all < 0.05). The cumulative one-year pregnancy rate for the M. hominis positive group was lower than that in the negative group (58.96% vs 66.76%, log-rank test: P = 0.029). After controlling for potential confounders, the risk of pregnancy in the M. hominis positive group was reduced by 38% when compared with the positive group (FOR = 0.62, 95% CI: 0.42-0.93). CONCLUSION: The vaginal colonization of M. hominis at a low level in pre-pregnant women is negatively correlated with female fecundability.

Exposure to Heavy Metals and Allergic Outcomes in Children: a Systematic Review and Meta-analysis.

It has been reported that heavy metals have adverse effects on the immune system. However, the relationship between heavy metal exposure and allergic outcomes remains unclear. This systematic review was conducted to examine whether heavy metal exposure is associated with allergic outcomes during childhood. We performed a systematic search of all relevant articles in Web of Science, EMBASE, and PubMed, from inception through to November 2020. We used odds ratio (OR) and the standard mean differences (SMDs) with 95% confidence intervals (CIs) to present estimates from individual studies. In addition, random-effects meta-analysis was used to pool the data. We also conducted the meta-regression and subgroup analysis to explore potential sources of heterogeneity. After duplicate removal, we finally included 35 articles in the systematic review and meta-analysis from an initial 11,181 articles. The overall results showed that copper (Cu) was associated with asthma (pooled SMD = 1.50, 95% CI = 0.13-2.86); in the subgroup analysis, the results indicated that lead (Pb) was associated with asthma (pooled OR = 6.27, 95% CI = 2.24-17.56), and Cu and Pb were connected with atopic dermatitis (SMD = - 1.05, 95% CI = - 1.45 to - 0.65; SMD = 5.68, 95% CI = 5.05-6.32), respectively. Mercury (Hg) was associated with atopic dermatitis (pooled OR = 1.13, 95% CI = 1.04-1.22) and wheeze (OR = 1.20, 95% CI = 1.05-1.37). The meta-analysis results indicate that Cu might be connected with childhood asthma, but not with other allergic diseases; Hg and Pb may have no association with allergic diseases during childhood. Given some limits observed in the current studies, more prospective cohort studies are still needed to verify our findings. Review registration: PROSPERO CRD42020222167.

Clinical Manifestations of Polycystic Ovary Syndrome and Associations With the Vaginal Microbiome: A Cross-Sectional Based Exploratory Study.

Background: Previous studies suggest that the vaginal microbiome is associated with polycystic ovary syndrome (PCOS). However, the clinical manifestations of PCOS are heterogeneous. Whether the vaginal microbiome is related with different clinical symptoms was unknown. Materials and Methods: In this cross-sectional study, 89 female patients with PCOS admitted to Zhongda Hospital (Nanjing, China) were included. Basic demographic information, health-related behaviors, clinical manifestations and sex hormone levels were comprehensively recorded for all patients. Vaginal swabs were acquired for microbiota sequencing of the V3-V4 region of the 16S rRNA gene. Results: The prevalence of bacterial vaginitis and vulvovaginal candidiasis was 15.7% and 13.5%, respectively, within the PCOS patients, which were the most important factors affecting the vaginal microbiome (permutational multivariate analysis of variance test, R2 = 0.108, P = 0.001). The vaginal microbiome was associated with specific clinical manifestations of PCOS, including acanthosis nigricans, intermenstrual bleeding, pregnancy history, testosterone level and anti-müllerian hormone level, with P values < 0.05. The abundance of Lactobacillus crispatus was higher (P = 0.010) while that of Lactobacillus iners was lower (P = 0.036) among PCOS patients with elevated testosterone levels. Other potential bacterial biomarkers were not statistically significant after adjusting for confounding factors. No evidence of associations of other common manifestations of PCOS, such as obesity and acne, with the vaginal microbiome was obtained. Conclusion: Vaginal bacterial species among PCOS patients with variable clinical manifestations, especially differences in testosterone levels, are distinct. Further studies are essential to investigate the microbiota and molecular mechanisms underpinning this disease.

Male reproductive toxicity involved in spermatogenesis induced by perfluorooctane sulfonate and perfluorooctanoic acid in Caenorhabditis elegans.

As a persistent organic pollutant, perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) have gained increasing research attention over recent years because of their potential risk to humans and the environment. In this paper, we investigated the reproductive toxicity of these pollutants using a C. elegans model to evaluate spermatogenesis throughout the entire developmental cycle of him-5 mutant by exposing to 0.001, 0.01, and 0.1 mmol/L PFOS or PFOA for 48 h. Experimental results suggested that PFOS and PFOA exposure led to reductions in brood size, germ cell number, spermatid size, and motility, and increases in rate of malformation spermatids. Analysis of variance (ANOVA) showed that exposure to PFOS resulted in higher levels of damage than PFOA in germ cells only in 0.001 mmol/L exposure group. RT-qPCR was used to further investigate the expression of genes associated with different stages of spermatogenesis, such as mitosis and meiosis, fibrous body-membranous organelles (FB-MOs), and sperm activation. The expression levels of wee-1.3, spe-4, spe-6, and spe-17 genes were increased, while those of puf-8, spe-10, fer-1, swm-1, try-5, and spe-15 genes were decreased. Our results suggesting that PFOS or PFOA may cause spermatogenesis damage by disrupting the mitotic proliferation, meiotic entry, formation of the MOs, fusion of the MOs and plasma membrane (PM), and pseudopods. Loss-of-function studies using puf-8 and spe-10 mutants revealed spe-10 gene was specifically involved in PFOS- or PFOA-induced reproductive toxicity via regulating one or more critical palmitoylation events, while puf-8 gene was not direct target of PFOS and PFOA, and PFOS and PFOA may act on the upstream gene of puf-8, thus affecting reproductive ability. Taken together, these results demonstrate the potential adverse impact of PFOS and PFOA exposure on spermatogenesis and provide valuable data for PFC risk assessment. Grapical abstract.

Serum Trace Elements in Patients With Polycystic Ovary Syndrome: A Systematic Review and Meta-Analysis.

Polycystic ovary syndrome (PCOS) is reported to be associated with certain trace elements. However, previous data are inconsistent and potentially biased due to small sample sizes. The potential utility of trace element levels for screening of PCOS remains to be established. The aim of this meta-analysis was to investigate the potential relationships between PCOS and serum levels of zinc (Zn), copper (Cu), magnesium (Mg), iron (Fe) and ferritin. We carried out a literature search of PubMed, EMBASE, and Web of Science for relevant cross-sectional/case-control studies published prior to October 2019. Random-effect models were used to estimate the overall standard mean differences (SMDs) between PCOS and healthy control subjects. The screening value of potential microelement biomarkers for PCOS was assessed using the receiver operating characteristic (ROC) curve. Twenty-one studies featuring 2,173 women with PCOS and 1,897 healthy women were selected for analysis. Our results showed that Cu and ferritin levels were significantly higher in women with PCOS than healthy controls, with SMDs of 0.52 [95% confidence interval (CI): 0.38-0.67, I2 = 47.6%] and 1.05 (95% CI: 0.25-1.86, I2 = 97.0%), respectively. The serum ferritin concentration was distinguished as a potential biomarker for PCOS based on the high area under ROC curve value of 0.71 (95% CI: 0.57-0.86). Although we did not identify a statistical association between serum Zn concentration and PCOS overall, the concentration of Zn in PCOS women with insulin resistance (IR) was lower than that in healthy women (SMD = -0.89, 95% CI: -1.73 to -0.06). Furthermore, the concentrations of Mg (SMD = 0.31, 95% CI: -0.32-0.94, I2 = 95.4%) and Fe (SMD = -0.59, 95% CI: -1.29-0.12, I2 = 97.2%) were not statistically significant between the PCOS and control groups. We generated hypothetical pathways for associations among serum Cu, ferritin and PCOS. The serum concentrations of both Cu and ferritin were significantly higher in women with PCOS, and ferritin was identified as a potential early indicator for PCOS screening. Further studies are essential to determine the specific underlying mechanisms.

Non-targeted metabolomic profiling of atrazine in Caenorhabditis elegans using UHPLC-QE Orbitrap/MS.

The widespread use of the herbicides Atrazine (ATR) has been raised attention due to its ubiquitous occurrence in the environment. As an endocrine disruptor, ATR causes reproductive, immune, nervous system toxicity in biota. In this study, we aimed to investigate metabolic profile characteristics and potential metabolic biomarker that reflects specific damage in toxic effect after ATR exposure. Hence, a metabolomics study was performed to determine the significantly affected metabolites and the reproduction and locomotion of C. elegans were investigated. Mediation analysis was used to evaluate the mediating effect of metabolites on association between ATR exposure and toxic effect. ATR (≥0.04 mg/L) caused the significant dose dependent reduction of brood size and locomotion behavior, however, the body length and width were significantly decreased only in 40 mg/L group. These results suggesting that brood size, head thrashes and body bends are more sensitive indictor to assessment ATR toxicity in C. elegans. Meanwhile, metabolomics analysis revealed that ATR exposure can induce metabolic profiles significant alterations in C. elegans. We found that 9 metabolites significantly increased and 18 metabolites significantly decreased, such as phosphatidylcholine, GMP, CDP-choline, neopterin etc. Those alteration of metabolites were mainly involved in the pathways: glycerophospholipid metabolism, glycolysis/gluconeogenesis, folate biosynthesis, glycine, serine and threoninemetabolism, pyrimidine and purine metabolism. Overall, these changes are signs of possible oxidative stress and ATP synthesis disruption modification. Mediation analysis showed a significant indirect effect of ATR exposure on brood size, via 7,8-dihydroneopterin 2',3'-cyclic-p, and phosphatidylcholine might mediate association between ATR exposure and body bends, suggesting that 7,8-dihydroneopterin 2',3'-cyclic-p and phosphatidylcholine might be potentially specificity marker for brood size and body bend respectively. This preliminary analysis investigates metabolic characteristics in C. elegans after ATR exposure, helping to understand the pathways involved in the response to ATR exposure and provide potential biomarkers for the safety evaluation of ATR.

The association between vaginal microbiota and female infertility: a systematic review and meta-analysis.

PURPOSE: This study aimed to explore the association between vaginal microbiota and infertility. METHODS: We searched a range of electronic databases for appropriate articles, including PubMed, Web of Science, Embase, Chinese National Knowledge Infrastructure (CNKI), and Wanfang, from inception to 8th September 2019. Identified articles were then screened using strict inclusion and exclusion criteria. By referring to Tamarelle's method, we divided vaginal microbiota into two categories: low-Lactobacillus vaginal microbiota (LL-VMB) and high-Lactobacillus vaginal microbiota (HL-VMB). Patients were defined as HL-VMB if they had a Nugent score of 0-3, a negative Amesel/Spiegel's test, or if the vaginal community status was dominated by either L. crispatus, L. iners, L. gasseri and L. jensenii via 16S rRNA sequencing. Otherwise, cases were regarded as LL-VMB. Statistical analyses were performed with STATA 13.0 statistical software. Effect estimates are presented as odds ratios (ORs) with 95% confidence intervals (CIs). RESULTS: Fifteen articles were included in our final analysis. The HL-VMB was negatively related to infertility; a fixed model showed that the pooled OR was 0.83 (95% CI 0.77-0.90). There was no significant publication bias, as determined by Begg's test (P = 0.488) and Egger's test (P = 0.652). Using a random effect model, the pooled OR for intermediate bacterial vaginitis (BV) and infertility was 1.39 (95% CI 1.10-1.76) and the pooled OR for positive BV was 1.72 (95% CI 1.10-2.69). Subgroup and sensitivity analyses further demonstrated that the associations identified were stable. However, the acquired evidence was not sufficient to make inferences with regards to the mechanisms underlying these relationships. CONCLUSION: This systematic review and meta-analysis identified a negative correlation between HL-VMB and female infertility. However, due to a variety of limitations, the evidence acquired does not allow us to identify the specific mechanisms underlying this association. Further high-quality studies are needed to verify the causal relationship and explore the molecular mechanisms involved.

Characteristics of the vaginal microbiome in cross-border female sex workers in China: a case-control study.

BACKGROUND: Female sex workers (FSWs) are key groups in the transmission of sexual transmitted infections (STI), and vaginal microbiome variations play an important role in transmission. We aimed to explore the characteristics of vaginal microbiome among FSWs. MATERIALS AND METHODS: A total of 24 cross-border FSWs were randomly selected from a cross-sectional survey for female sex workers in southwest China. Thirty-seven female non-sex workers (FNSWs) were randomly selected from the gynecology clinic and health examination center. Vaginal swabs were collected, bacterial DNA extracted and 16S rRNA genes were sequenced. Differences in the vaginal microbiome between both groups were compared using bioinformatics analysis. RESULTS: One DNA sample was excluded due to unqualified concentration, therefore 60 samples were sequenced. FSWs had significantly different vaginal microbiota ß diversity, but undifferentiated α diversity when compared with non-sex workers. The average relative abundance of Sneathia, Shigella, Neisseria, Chlamydia, Prevotella, Enterococcus and Ureaplasma among FSWs was higher than FNSWs, and relative abundance of Atopobium in FSWs was lower than FNSWs. The Lactobacillus genus was the major genus in both groups. At the species level, Lactobacllus crispatus, Lactobacllus gasseri and Lactobacllus jensenii, in female sex workers, were lower when compared to FNSWs. CONCLUSION: There were distinct differences in vaginal bacteria variety between FSWs and FNSWs. Some disease-related genus were also more abundant in FSWs. Based on these observations, further research is required to identify microbiome communities related to high STI risks and other diseases in these cohorts.

Novel-miR-4885 Promotes Migration and Invasion of Esophageal Cancer Cells Through Targeting CTNNA2.

Esophageal cancer is one of the most common cancers worldwide. It is critical to find early diagnostic biomarkers for esophageal cancer. MicroRNAs (miRNAs) play important regulatory roles in occurrence and development of esophageal cancer, which has the diagnostic and prognostic values. The aim of the study was to evaluate the potential diagnostic value of the novel miRNA. The novel miRNA was predicted using miRDeep2 software and validated by quantitative reverse transcription PCR (qRT-PCR) and the TA-cloning sequencing of the PCR products. The expression of the novel miRNA in esophageal cancer tissues and adjacent tissues was also analyzed by qRT-PCR. The EdU staining and transwell method were used to detect the capacity of cell proliferation, migration, and invasion. Besides, the target gene CTNNA2 of novel-miR-4885 was verified via qRT-PCR, western blot, luciferase reporter assay, and RNA immunoprecipitation. We identified the novel-miR-4885, the expression level was confirmed by qRT-PCR in the esophageal cancer cells. The result of TA-cloning sequencing was consistent with the prediction and the pre-miRNA had a standard hairpin stem-loop structure. In addition, the expression of novel-miR-4885 was upregulated in esophageal cancer tissue compared with that in adjacent tissue (p < 0.05). Further, the assays showed that overexpression novel-miR-4885 could improve the cell proliferation, migration, and invasion with an average fold change of 1.19, 1.59, and 2.34, respectively. Novel-miR-4885 can bind to 3' untranslated region of CTNNA2 to reduce cell adhesion and promote epithelial-mesenchymal transition in esophageal cancer cell. The expression of N-cadherin, ß-catenin, Vimentin, and α-smooth muscle actin was upregulated, while that of E-cadherin and ZO-1 proteins was downregulated through western blot. Novel miRNA present in esophageal cancer cells was validated, supplementing the miRNA database. Meantime, the possible functional mechanisms were explored, and the results showed that the novel miRNA may serve as potential biomarker for the diagnosis of esophageal cancer.

Di (2-ethylhexyl) phthalate-induced reproductive toxicity involved in dna damage-dependent oocyte apoptosis and oxidative stress in Caenorhabditis elegans.

Di-(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer with a high environmental exposure level. As a persistent organic pollutant, DEHP causes reproductive and developmental toxicity in mammals. In this paper, the reproductive toxicity of DEHP was discussed using the model organism Caenorhabditis elegans to determine the sensitivity indices for evaluating the ecotoxicological effects of DEHP. L4 C. elegans larvae to evaluate the LC50 of DEHP and the changes in brood size and generation time, we found that the LC50 of DEHP to C. elegans exceeded 100 mg/L. And 10 mg/L DEHP exposure significantly reduced the brood sizes but not the generation time. Results of oocyte and distal-tip cell (DTC) counting suggested that the number of oocytes were decreased and apoptotic cells that from the unilateral gonad arm were increased in the 1 mg/L and 10 mg/L DEHP exposed groups. In contrast, there was no significant difference in the fluorescence intensity of DTC. Fluorescence analysis of HUS-1 showed that HUS-1 protein was overexpressed after DEHP exposure. The H2O2 level and DNA damage were measured by Bradford protein assay and AP staining respectively. The results showed that there was no significant difference in H2O2 level after DEHP exposure, in contrast, DNA damage was increased significantly. Moreover, 10 mg/L concentration DEHP exposure significantly increased the expression levels of apoptosis-related genes cep-1, egl-1, ced-4, and ced-3 and decreased the expression levels of ced-9. It suggested that cep-1, egl-1, ced-4, and ced-3 genes promote apoptosis and the ced-9 gene inhibits apoptosis. Meanwhile, 10 mg/L concentration DEHP exposure decreased the expression of oxidative stress-related genes mev-1 and gas-1. The mev-1 and gas-1 are mainly involved in the inhibition of oxidative stress in nematodes. In short, the decreased oocyte numbers and increased apoptosis oocyte numbers in C. elegans when exposed to DEHP, which may involve in the DNA damage induced by oxidative stress.

Novel combined miniature optical coherence tomography ultrasound probe for in vivo intravascular imaging.

We have developed a miniature integrated optical coherence tomography (OCT) ultrasound (US) probing system for intravascular imaging applications. In the OCT probe, the light coming out of a single mode fiber is focused by a gradient-index lens and then reflected by a right-angle prism from the side of the probe into the sample. It was combined with a 35 MHz PMN-PT side-viewing ultrasound transducer to obtain the ultrasound image as well. The OCT and ultrasound probes were integrated as a single probe to obtain OCT and ultrasound images simultaneously. The integrated probe has an outer diameter of 0.69 mm which, to our knowledge, is the smallest integrated OCT-US probe reported. Fast data acquisition and processing was implemented for real-time imaging. In vitro OCT and US images of human coronary artery with pathology, as well as in vivo images of normal rabbit abdominal aorta, were obtained using the integrated OCT-US probe to demonstrate its capability.

Quantification of airway thickness changes in smoke-inhalation injury using in-vivo 3-D endoscopic frequency-domain optical coherence tomography.

Smoke inhalation injury is frequently accompanied by cyanide poisoning that may result in substantial morbidity and mortality, and methods are needed to quantitatively determine extent of airway injury. We utilized a 3-D endoscopic frequency-domain optical coherence tomography (FD-OCT) constructed with a swept-source laser to investigate morphological airway changes following smoke and cyanide exposure in rabbits. The thickness of the mucosal area between the epithelium and cartilage in the airway was measured and quantified. 3-D endoscopic FD-OCT was able to detect significant increases in the thickness of the tracheal walls of the rabbit beginning almost immediately after smoke inhalation injuries which were similar to those with combined smoke and cyanide poisoning.

A dual-modality probe utilizing intravascular ultrasound and optical coherence tomography for intravascular imaging applications.

We have developed a dual-modality biomedical imaging probe utilizing intravascular ultrasound (IVUS) and optical coherence tomography (OCT). It consists of an OCT probe, a miniature ultrasonic transducer and a fixed mirror. The mirror was mounted at the head of the hybrid probe 45° relative to the light and the ultrasound beams to change their propagation directions. The probe was designed to be able to cover a larger area in blood vessel by IVUS and then visualize a specific point at a much finer image resolution using OCT. To demonstrate both its feasibility and potential clinical applications, we used this ultrasound-guide OCT probe to image a rabbit aorta in vitro. The results offer convincing evidence that the complementary natures of these two modalities may yield beneficial results that could not have otherwise been obtained.

High-resolution coregistered intravascular imaging with integrated ultrasound and optical coherence tomography probe.

We report an integrated ultrasound (US) and optical coherence tomography (OCT) probe and system for intravascular imaging. The dual-function probe is based on a 50 MHz focused ring US transducer, with a centric hole for mounting OCT probe. The coaxial US and light beams are steered by a 45° mirror to enable coregistered US∕OCT imaging simultaneously. Lateral resolution of US is improved due to focused ultrasonic beam. Mirror effects on US were investigated and invitro imaging of a rabbit aorta has been carried out. The combined US-OCT system demonstrated high resolution in visualizing superficial arterial structures while retaining deep penetration of ultrasonic imaging.

Integrated intravascular optical coherence tomography ultrasound imaging system.

We report on a dual-modality optical coherence tomography (OCT) ultrasound (US) system for intravascular imaging. To the best of our knowledge, we have developed the first integrated OCT-US probe that combines OCT optical components with an US transducer. The OCT optical components mainly consist of a single-mode fiber, a gradient index lens for light-beam focusing, and a right-angled prism for reflecting light into biological tissue. A 40-MHz piezoelectric transducer (PZT-5H) side-viewing US transducer was fabricated to obtain the US image. These components were integrated into a single probe, enabling both OCT and US imaging at the same time. In vitro OCT and ultrasound images of a rabbit aorta were obtained using this dual-modality imaging system. This study demonstrates the feasibility of an OCT-US system for intravascular imaging, which is expected to have a prominent impact on early detection and characterization of atherosclerosis.