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BPDE (benzo(a)pyren-7,8-dihydrodiol-9,10-epoxide), a metabolite of environmental carcinogenic BaP, weakens the migration and invasion of human villous trophoblast cells and may further induce miscarriage. However, the underlying mechanisms remain largely unknown. In this study, we identified that in trophoblast Swan 71 and HTR-8/SVneo cells, miR-hz02 upregulates the level of lnc-HZ02, which inhibits the expression of an RNA-binding protein HuR. HuR could interact with FAK mRNA and promote its mRNA stability, thus upregulating the FAK level and the FAK/SRC/PI3K/AKT pathway, and finally maintaining the normal migration and invasion of trophoblast cells. If trophoblast cells are exposed to BPDE, both miR-hz02 and lnc-HZ02 are upregulated, which reduce the level of HuR, weaken the interactions of HuR with FAK mRNA, downregulate FAK level and the FAK/SRC/PI3K/AKT pathway, and finally inhibit cell migration and invasion. This study provides a novel scientific understanding of the dysfunctions of human trophoblast cells.

Regulatory T cells (Treg) can impede antitumor immunity and currently represent a major obstacle to effective cancer immunotherapy. Targeting tumor-infiltrating regulatory Treg while sparing systemic Treg represents an optimal approach to this problem. Here, we provide evidence that the interleukin 23 receptor (IL23R) expressed by tumor-infiltrating Treg promotes suppressive activity. Disruption of the IL23R results in increased responsiveness of destabilized Treg to the IL12 cytokine, the production of γ-interferon, and the recruitment of CD8 T cells that inhibit tumor growth. Since the Treg destabilization pathway that is initiated by IL23R blockade is distinct and independent from the destabilization pathway coupled to glucocorticoid-induced TNFR-related protein (GITR) activation, we examined the impact of the coordinate induction of the two destabilization pathways on antitumor immune responses. Combined GITR and IL23R antibody treatment of mice inoculated with MC38 tumors resulted in robust and synergistic antitumor responses. These findings indicate that the delineation of independent Treg destabilization pathways may allow improved approaches to the development of combination immunotherapy for cancers.

The conventional semen parameter analysis is widely used to assess male fertility. However, studies have found that ~15% of infertile patients show no abnormalities in conventional semen parameters. Additional technologies are needed to explain the idiopathic infertility and detect subtle sperm defects. Currently, biomarkers of sperm function, including sperm apoptosis, mitochondrial membrane potential (MMP), and DNA damage, reveal sperm physiology at the molecular level and are capable of predicting male fertility. With flow cytometry (FCM) techniques, each of these markers can be rapidly, accurately, and precisely measured in human semen samples, but time costs substantially increase and results could be obstructed if all the biomarkers need to be tested with a single cytometer. In this protocol, after collection and immediate incubation at 37 °C for liquefication, semen samples were further analyzed for sperm apoptosis using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. The MMP was labeled with 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-benzimidazolylcarbocyanine iodide (JC-1) probe, and DNA damage was assessed using the sperm chromatin structure assay (SCSA) with acridine orange (AO) staining. Thus, flow cytometric analysis of sperm function markers can be a practical and reliable toolkit for the diagnosis of infertility and evaluation of sperm function at both bench and bed.

Background Hirschsprung disease (HSCR) is a rare congenital gastrointestinal disease characterized by the absence of intestinal submucosal and myometrial ganglion cells. Recently, researches indicated that miR-100 regulated the growth, differentiation and apoptosis of neurons, and affected the functions of HSCR-associated pathways. While miR-100 rs1834306 A>G polymorphism was shown to modify the susceptibility to tumors, the association between this polymorphism and HSCR susceptibility is still unknown. Methods This was a case–control study consisting of 1470 HSCR cases and 1473 controls from southern China. DNA was genotyped by TaqMan real-time PCR. Odds ratios (ORs) and 95% confidence intervals (CIs) were used as statistical indicators. Results We found that miR-100 rs1834306 G allele and GG genotype significantly increased HSCR susceptibility (GG vs AA: adjusted OR=1.31, 95% CI=1.04–1.64, P=0.020; G vs A: adjusted OR=1.12, 95% CI=1.01–1.25, P=0.041; GG vs AA/AG: adjusted OR=1.30, 95% CI=1.07–1.59, P=0.010). In the stratified analysis, miR-100 rs1834306 GG genotype carriers had higher risk to develop HSCR in all clinical subtypes when compared with those with AA/AG genotypes, and OR was rising with HSCR aggravation (SHSCR: adjusted OR=1.28, 95% CI=1.03–1.59, P=0.029; LHSCR: adjusted OR=1.48, 95% CI=1.06–2.07, P=0.020; TCA: adjusted OR=2.12, 95% CI=1.22–3.69, P=0.008). Conclusion Our findings suggested that miR-100 rs1834306 A>G polymorphism was associated with increased HSCR susceptibility in southern Chinese children. Furthermore, miR-100 rs1834306 GG genotype had a greater genetic pathopoiesis in severe HSCR.

Five novel bisindole alkaloids, hunzeylanines A-E (1-5), with an unprecedented skeleton were isolated from the roots of Hunteria zeylanica. Compounds 1-5 represent the first examples of akuammine-pleioarpamine-type bisindole alkaloids fused with a dihydropyran unit. Their structures including absolute configurations were established through comprehensive spectroscopic data analyses and computational calculation methods. The plausible biogenetic pathway of 1 was also proposed. Alkaloids 1 and 2 displayed moderate cytotoxicity toward three human cancer cell lines (MDA-MB-231, AV3, and Huh7).

STUDY QUESTION Is circadian desynchrony a risk factor of male reproductive damage in semen parameters and/or reproductive hormones? SUMMARY ANSWER Circadian desynchrony correlates with decrease of sperm count, which was improved when circadian desynchrony was attenuated. WHAT IS KNOWN ALREADY Circadian desynchrony caused by work (shift work) and non-work-related reasons is prevalent worldwide and has been found to be associated with decreased female fertility, but whether it harms male reproductive health is unclear. STUDY DESIGN, SIZE, DURATION A hybrid research was conducted. (i) A cross-sectional study of 1346 Chinese men in 2007 was used to analyze the association between semen/hormone biomarkers and work-related circadian desynchrony, which was divided into rotating shift work and permanent shift work against non-shift work. (ii) A cohort of 796 Chinese undergraduates from 2013 to 2014 was used to analyzed the association between semen/hormone biomarkers and non-work-related circadian desynchrony (between school days and days off). (iii) The biomarker identified simultaneously in both populations was further validated in male C57BL/6J mice housed under conditions simulating circadian desynchrony. PARTICIPANTS/MATERIALS, SETTING, METHODS A total of 17 semen/hormone biomarkers were compared among rotating shift workers and permanent shift workers against non-shift workers in the 1346 reproductive-age Chinese men. A total of 14 semen/hormone biomarker was analyzed in the undergraduate cohort for correlation with non-work-related circadian desynchrony (measured by Munich Chronotype Questionnaire) in 2013 and 2014 and compared between the 2 years. Photoperiod-shifting method was used to establish the mouse model, in which the biomarker was examined and molecular mechanism was explored by apoptosis analysis, DNA content analysis, transcriptome sequencing, real-time PCR and western blotting. MAIN RESULTS AND THE ROLE OF CHANCE Among the semen/hormone biomarkers, sperm count was found to be lower in rotating shift workers, who had a higher risk of low sperm count defined by Chinese Ministry of Health (total sperm/ejaculate < 120 × 106) than non-shift workers (odds ratio = 1.26, 95% CI 1.05–1.52). This biomarker was replicated in the undergraduate cohort, where each hour of circadian desynchrony was associated with 1.16 (95% CI 1.02–1.31) fold odds of low sperm count, and sperm count increased during 2014 in men who reduced circadian desynchrony after 2013. A decrease of sperm count with circadian desynchrony and its recovery after removal of circadian desynchrony was also observed in the mouse model. During asynchrony, increased apoptosis was found in seminiferous tubules and the marker genes of post-spermatocyte stage cells were down-regulated. The most enriched functional pathway was homologous recombination, which happened during meiosis. LIMITATIONS, REASONS FOR CAUTION The study of human beings was observational while the animal study has potential difference in circadian desynchrony exposure and species susceptibility. Further researches are needed to clarify the causal relationship in men. WIDER IMPLICATIONS OF THE FINDINGS These findings provide novel insight to the effect of circadian desynchrony on male reproductive health and a potential strategy for prevention of reproductive damage. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the National Key R&D Program of China [2017YFC1002001] and National Natural Science Foundation of China [81871208]. There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER NA.

Strand displacement DNA synthesis is essential for DNA replication. Gp90, the sole DNA polymerase of Pseudomonas aeruginosa phage 1, can bypass multiply DNA lesions. However, whether it can perform strand displacement synthesis is still unknown. In this work, we found that gp90 exo- could perform strand displacement synthesis, albeit its activity and processivity were lower than those of primer extension. Gp90 exo- itself could not unwind Y-shaped or fork DNA. Tail and gap at DNA fork were necessary for efficient synthesis. High GC content obviously inhibited strand displacement synthesis. Consecutive GC sequence at the entrance of fork showed more inhibition effect on DNA synthesis than that in the downstream DNA fork. The fraction of productive polymerase and DNA complex (A values) was higher for fork than gap; while their average extension rates (kp values) were similar. However, both A and kp values were lower than those for the primer/template (P/T) substrate. The binding of gp90 exo- to fork was tighter than P/T or gap in the absence of dATP. In the presence of dATP to form ternary complex, the binding affinity of gp90 exo- to P/T or gap was increased compared with that in the binary complex. Abasic site, 8-oxoG, and O6-MeG inhibited and even blocked strand displacement synthesis. This work shows that gp90 exo- could perform strand displacement DNA synthesis at DNA fork, discovering the presence of new functions of PaP1 DNA polymerase in DNA replication and propagation of PaP1.

Two novel monoterpenoid indole alkaloids, alstolarines A and B (1–2), were isolated from the leaves and twigs of Alstonia scholaris. Compound 1 is the first vobasinyl-type alkaloid with a 6/5/8/5/6/5 hexacyclic architecture featuring an unprecedented 21-oxa-4-azaspiro[4,4]nonane unit. Compound 2 is the first 21-nor-vobasinyl-type alkaloid with a unique 3-oxa-4-azatricyclo[5.1.1.13,20]undecane ring system. Their structures including absolute configurations were established by a combination of spectroscopic data analyses, electronic circular dichroism (ECD) calculations, and single crystal X-ray diffraction analysis. The plausible biogenetic pathways of 1 and 2 were also proposed. Compounds 1 and 2 exhibited promising vasorelaxant activities with EC50 values of 4.6 and 6.7 μM, and compound 2 showed moderate acetylcholinesterase (AChE) inhibitory activity with an IC50 value of 19.3 μM.

SUMMARY Follicular regulatory T (TFR) cells are a specialized suppressive subset that controls the germinal center (GC) response and maintains humoral self-tolerance. The mechanisms that maintain TFR lineage identity and suppressive activity remain largely unknown. Here, we show that expression of Blimp1 by FoxP3+ TFR cells is essential for TFR lineage stability, entry into the GC, and expression of regulatory activity. Deletion of Blimp1 in TFR cells reduced FoxP3 and CTLA-4 expression and increased pro-inflammatory cytokines and spontaneous production of autoantibodies, including elevated IgE. Maintenance of TFR stability reflected Blimp1-dependent repression of the IL-23R-STAT3 axis and activation of the CD25-STAT5 pathway, while silenced IL-23R-STAT3 or increased STAT5 activation rescued the Blimp1-deficient TFR phenotype. Blimp1-dependent control of CXCR5/CCR7 expression also regulated TFR homing into the GC. These findings uncover a Blimp1-dependent TFR checkpoint that enforces suppressive activity and acts as a gatekeeper of GC entry., In Brief Wang et al. identify Blimp1 as a critical transcription factor for the proper positioning and stable expression of the suppressive activity of TFR cells that control GC responses. In the absence of Blimp1, unstable TFR cells prematurely migrate into the GC and differentiate into TFH-like cells to promote dysregulated GC responses., Graphical Abstract

The abnormal accumulation of α-synuclein (α-syn) is a crucial factor for the onset and pathogenesis of Parkinson’s disease (PD), and the autophagy-lysosome pathway (ALP) contributes to α-syn turnover. AMP-activated protein kinase (AMPK) and the mammalian target of rapamycin (mTOR) regulate autophagy by initiating the macroautophagy cascade and promoting lysosomal biogenesis via increased transcription factor EB (TFEB) activity. Hence, activation of AMPK-mTOR-TFEB axis-mediated autophagy might promote α-syn clearance in PD. Harmol is a β-carboline alkaloid that has been extensively studied in a variety of diseases but rarely in PD models. In this study, we aimed to evaluate the effect and underlying mechanism of harmol in PD models in vitro and in vivo. We show that harmol reduces α-syn via ALP in a dose- and time-dependent manner in cell model that overexpressed human A53T mutant α-syn. We also demonstrate that harmol promotes the translocation of TFEB into the nucleus and accompanies the restoration of autophagic flux and lysosomal biogenesis. Importantly, harmol improves motor impairment and down-regulates α-syn levels in the substantia nigra and prefrontal cortex in the α-syn transgenic mice model. Further studies revealed that harmol might activate ALP through AMPK-mTOR-TFEB to promote α-syn clearance. These in vitro and in vivo improvements demonstrate that harmol activates the AMPK-mTOR-TFEB mediated ALP pathway, resulting in reduced α-syn, and suggesting the potential benefit of harmol in the treatment of PD.

It is controversial whether exposure to isoflavones exerts male reproductive toxicity. The aim of this study was to investigate whether isoflavone exposure during adulthood could have deleterious impacts on male reproductive health by the cross-sectional study, animal experiments, and in vitro tests. In the cross-sectional study, we observed that urinary isoflavones were not significantly associated with semen quality including sperm concentrations, sperm count, progressive motility, and total motility, respectively. However, negative associations were found between plasma testosterone and urinary Σisoflavones, genistein, glycitein, and dihydrodaidzein. In the animal experiments, serum and intratesticular testosterone levels were decreased in mice exposed to several dosages of genistein. Genistein administration caused upregulation of estrogen receptor alpha and downregulation of cytochrome P45017A1 protein levels in testes of mice. In vitro tests showed that genistein caused a concentration-dependent inhibition of testosterone production by TM3 Leydig cells. Elevated protein expression of estrogen receptor alpha and decreased messenger RNA/protein level of cytochrome P45017A1 were also observed in genistein-treated cells. Protein level of cytochrome P45017A1 and testosterone concentration were significantly restored in the estrogen receptor alpha small interferring RNA-transfected cells, compared to cells that treated with genistein alone. The results demonstrate that exposure to isoflavones during adulthood may be associated with alterations of reproductive hormones. Particularly, genistein, which inhibits testosterone biosynthesis through upregulation of estrogen receptor alpha in Leydig cells of mice, might induce the disruption of testosterone production in human. The present study provides novel perspective into potential targets for male reproductive compromise induced by isoflavone exposure.

This study aimed to investigate the situations of pornography use among male college students of China, to explore the addiction possibility for pornography use, and to study the associations between pornography use and reproductive hormone levels and semen quality. Five hundred sixty-eight participants met the inclusion criteria and finished all of the questionnaires and hormone level and semen parameter examinations. A majority of participants (except one) had pornography use experience, 94.2% participants started pornography use before college, and 95.9% participants reported they had masturbation experience when using pornography. Early contact to pornography, frequent pornography use, high amount of time spending on pornography use, and frequent masturbation during pornography use were correlated with addiction trends. Earlier pornography use was found to be associated with lower serum prolactin (PRL), follicle-stimulating hormone (FSH), and progesterone (Prog), as well as lower sperm concentration and total sperm count. Higher frequency of pornography use was associated with lower serum estrogen (E2). In conclusion, pornography use was common among male college students in China. Early contact, high frequent use, and high frequency of masturbation during pornography use could lead to addiction trends and aberrant reproductive hormone levels and semen quality.

ObjectivesTo explore the role of folate metabolism in 1,3-Butadiene (BD)'s genotoxicity, we conducted a match-up study in BD-exposed workers in China to analyze the associations between the polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and the chromosomal damage induced by BD exposure, and culture-based experiments in TK-6 cells to examine the global DNA methylation levels and chromosomal damage when exposed both to BD’s genotoxic metabolite, 1,2:3,4-diepoxybutane (DEB), and MTHFR’s direct catalytic product, 5-methyltetrahydrofolate (5-MTHF).MethodsCytokinesis block micronucleus assay (CBMN) was used to examine the chromosomal damage induced by BD or DEB. Poisson regression models were produced to quantify the relationship of chromosomal damage and genetic polymorphisms in the BD-exposed workers. Global DNA methylation levels in TK6 cells were examined using DNA Methylation Quantification Kit.ResultsWe found that BD-exposed workers carryingMTHFRC677T CC (2.00 ± 2.00‰) (FR = 0.36, 95%CI: 0.20–0.67,P MTHFRC677T CT (2.87 ± 1.98‰) (FR = 0.49, 95%CI: 0.32–0.77,P MTHFR677 TT (5.33 ± 2.60‰), respectively. The results in TK6 cells showed that there was a significant increment in frequencies of micronucleus (MN), nucleoplasmic bridge (NPB) and nuclear bud (NBUD) with exposure to DEB at each 5-MTHF dose (ANOVA,P P P P ConclusionWe reported that folate metabolism could modify the association between BD exposure and chromosomal damage, and such effect may be partially mediated by DNA hypomethylation, and 5-MTHF supplementation could rescue it.

Male fertility has shown a continuously declining tendency for decades. Over exposure to metal/metalloid elements has been proposed as associated with reproductive impairment. However, the hazard profile remained unclear in general public experiencing low-level and combined metal exposure.Based on the MARHCS cohort in Chongqing, China, 796 college students were recruited from June 2013 and 666 subjects were followed up next year. At each phase, semen and blood samples were collected for an assessment of semen quality and six sex hormones levels. Eighteen urinary metal/metalloid elements were quantified by ICP-MS as internal exposure biomarkers. Cluster analysis was conducted to characterize reproductive outcomes in the subgroups for different overall estimated exposure levels. Effects of each metal/metalloid element were analyzed using multiple statistical strategies: single-element mixed model, multiple-elements model and self before-after comparison design.The urine concentration for 18 metal/metalloid elements was at a typically lower level (far away from the exposure limits) and positively associated with each other. After adjustment of the potential confounders, a decrease of 11.53% (95% CI: -18.61, -3.84%) and 10.84% (95% CI: -17.93, -3.14%) in spermatid morphology was observed in the highest quantile groups of vanadium (V) and nickel (Ni), respectively. Urinary silver (Ag) was dose-dependent associated with an increase in total sperm number (6.91%, 95% CI: 1.14, 13.00%), sperm concentration (16.38%, 95% CI: 5.15, 28.81%) and semen volume (23.73%, 95% CI: 10.46, 38.60%). Further, hormone testosterone presented a significant decrease in subgroup with higher overall estimated exposure and a stable negative association with lithium (Li). The above relationships remained significant across different statistical strategies (all p values0.05).Our study provided new evidences that exposure to metal/metalloid elements potentially exert bidirectional influences on semen quality at a relatively low level. And serum testosterone appears as a vulnerable index for metal exposure.

Three new monoterpenoid indole alkaloids (MIAs), hunterines A-C (1-3), were isolated from Hunteria zeylanica. Compound 1 possesses a unique skeleton with an unprecedented azabicyclo[4.3.1]decane ring system. Compounds 2 and 3 are a pair of epimeric vobasinylindole alkaloid heterodimers. Their structures including absolute configurations were established by spectroscopic analyses, X-ray diffraction, computational calculation, and the modified Mosher's method. Plausible biogenetic pathways of 1-3 were also proposed. Alkaloid 1 showed moderate cytotoxic activity against the HepG2 cell line.

Background Neuroblastoma is one of the most common extracranial solid pediatric tumors. KRAS plays an important role in regulating cell proliferation, differentiation, and apoptosis. Single nucleotide polymorphisms (SNPs) in KRAS have been shown to modify susceptibility to multiple tumors, but no specific molecular epidemiology study was reported regarding neuroblastoma. Methods We conducted a four-center case-control study to explore the association between KRAS gene polymorphisms (rs12587 G>T, rs7973450 A>G, rs7312175 G>A) and neuroblastoma susceptibility with 505 Chinese children and 1070 matched controls. Results We found that rs7973450 A>G was associated with significantly increased neuroblastoma risk [GG vs. AA: adjusted odds ratio (OR)=4.26, 95% confidence interval (CI)=1.28–14.19, P=0.018; GG vs. AA/AG: adjusted OR=4.27, 95% CI=1.28–14.24, P=0.018]. The stratified analysis further demonstrated that rs7973450 GG genotype carriers had a higher risk to develop neuroblastoma in the subgroups of males, tumor originated from the adrenal gland and clinical stages III+IV. Conclusions Overall, our results suggested that rs7973450 A>G was associated with increased neuroblastoma risk.

Dibutyl phthalate (DBP)-induced germ cell apoptosis contributes to male reproductive toxicity, however, the primary target organelle of DBP or the molecular events triggered by DBP to initiate germ cell apoptosis remain unclear. Our previous studies demonstrated DBP could stimulate the production of intracellular reactive oxygen species (ROS), which served as an upstream mediator of activation of endoplasmic reticulum (ER) stress in mouse spermatocyte-derived GC-2 cells. In the present study, the impacts of DBP-induced ROS generation on the mitochondria-related damage and the associations between ER stress and mitochondrial-related damage were investigated in GC-2 cells. We observed significant decreases of mitochondrial mass, mtDNA copy number, COX IV protein level, and ATP level in DBP-treated GC-2 cells in a dose-dependent manner. And DBP activated mitochondrial-related apoptosis, indicated by the elevation of cytoplasmic cytochrome C (Cyt C) and the activation of caspase-9/3 cascade. Pretreatment with antioxidant melatonin obviously attenuated DBP-induced mitochondrial damage and mitochondrial-dependent apoptosis in GC-2 cells, indicating the role of ROS in DBP-caused testicular toxicity. In response to oxidative stress, the Nrf2/ARE axis was activated in DBP-treated GC-2 cells to counteract ROS overproduction and subsequent mitochondrial damage. Further experiments showed DBP treatment increased the phosphorylated expression of ER stress-related protein PERK. GSK2606414, a specific inhibitor of PERK, partly attenuated the expression of Nrf2. And both DBP-induced mitochondrial damage in GC-2 cells and mitochondrial-dependent apoptosis of the germ cells in rat testes were further aggravated by PERK inhibition. Taken together, our data suggest that PERK regulates the Nrf2/ARE antioxidant pathway functioning as a self-defense mechanism against ROS-related mitochondrial damage induced by DBP in male germ cells.

Although TC2N has proven to be an oncogene in lung cancer, its biological function and molecular mechanisms in other cancer still remains unclear. Here, we investigate in breast cancer that TC2N expression is sharply overexpressed in breast cancer specimens compared with normal breast specimens, and the low TC2N expression was associated with advanced stage, lymphatic metastasis, larger tumors and shorter survival time. Upregulation of TC2N significantly restrains breast cancer cell proliferation in vitro and tumor growth in vivo. Mechanistically, TC2N blocks AKT signaling in a PI3K dependent and independent way through weakening the interaction between ALK and p55γ or inhibiting the binding of EBP1 and AKT. To sum up, these results unmask an ambivalent role of TC2N in cancer, providing a promising inhibitor for PI3K-AKT signaling.

The present study aimed to investigate the potential inhibitory effects of 21 clinically available hypnotics on acetylcholine (ACh)-induced contractions in rat urinary bladder smooth muscle (UBSM) in order to predict whether these hypnotics could induce voiding impairment. ACh-induced contraction in rat UBSM was inhibited only by diphenhydramine (a histamine H1 receptor antagonist) at a concentration that was clinically relevant. ACh-induced contraction was also significantly inhibited by flurazepam (a benzodiazepine hypnotic) and suvorexant (an orexin receptor antagonist), albeit at concentrations that substantially exceeded clinically achievable blood levels. These three drugs (at 10-5 M) also inhibited high-KCl (80 mM) Locke-Ringer solution-induced contractions. In contrast to the effects of the abovementioned hypnotics, ACh-induced contractions were not significantly affected by triazolam, etizolam, brotizolam, lormetazepam, estazolam, flunitrazepam, nitrazepam (benzodiazepine hypnotics), thiopental, thiamylal, pentobarbital, amobarbital, secobarbital, phenobarbital (barbiturate hypnotics), zolpidem (an imidazopyridine hypnotic), zopiclone (a cyclopyrrolone hypnotic), ramelteon (a melatonin receptor agonist), bromovalerylurea, and chloral hydrate. These findings suggest that most clinically used hypnotics are not likely to result in anticholinergic-induced dysuria within their clinically achievable blood concentration ranges. Diphenhydramine may, however, induce voiding impairment, an action attributable to diminished UBSM contractility within its clinical dose range.

Background: A report examining whether clinically available antidepressants increase urethral smooth muscle contraction via antagonistic effects on the α2-adrenoceptor (α2-AR) is lacking. Objectives: The present study was performed to evaluate the potential of clinically available antidepressants to reverse α2-AR-mediated contractile inhibition in rat vas deferens, in order to predict whether they can induce voiding impairment. Method: The effects of 18 antidepressants of different classes on electrical field stimulation (EFS)-induced contractions suppressed by 10–8 mol/L clonidine (a selective α2-AR agonist) in isolated rat vas deferens were investigated and related to their respective clinical blood concentrations. Results: The EFS-induced contractions suppressed by clonidine were recovered by amitriptyline (a tricyclic antidepressant), mirtazapine (a noradrenergic and specific serotonergic antidepressant), and trazodone (a serotonin 5-HT2A receptor antagonist) at concentrations close to the clinical blood levels. EFS-induced contractions were also recovered by trimipramine, clomipramine (tricyclic antidepressants), mianserin (a tetracyclic antidepressant), sertraline (a selective serotonin reuptake inhibitor [SSRI]), and sulpiride (a dopamine D2-receptor antagonist), albeit at concentrations that substantially exceeded their clinically-achievable blood levels. EFS-induced contractions were not significantly affected by imipramine, nortriptyline, amoxapine (tricyclic antidepressants), maprotiline (a tetracyclic antidepressant), fluvoxamine, paroxetine, escitalopram (SSRIs), milnacipran, duloxetine (serotonin and noradrenaline reuptake inhibitors), and aripiprazole (a dopamine partial agonist). Conclusions: These findings suggest that amitriptyline, mirtazapine, and trazodone induce voiding impairment caused by increased urethral resistance by enhancing sympathetic nerve activities attributed to α2-AR antagonism.

Background Pulmonary arterial hypertension (PAH), characterized by pulmonary artery constriction and vascular remodeling, has a high mortality rate. New drugs for the treatment of PAH urgently need to be developed. Purpose This study was designed to investigate the vasorelaxant activity of OTNA in isolated pulmonary arteries, and explore its molecular mechanism. Methods Pulmonary arteries and thoracic aortas were isolated from mice, and vascular tone was tested with a Wire Myograph System. Nitric oxide levels were determined with DAF-FM DA and DAX-J2™ Red. Cellular thermal shift assays, microscale thermophoresis, and molecular docking were used to identify the interaction between OTNA and aryl hydrocarbon receptor (AhR). The levels of PI3K, p-PI3K, Akt, p-Akt, eNOS, p-eNOS, and AhR were analyzed by Western blotting. Results OTNA selectively relaxed the isolated pulmonary artery rings in an endothelium-dependent manner. Mechanistic study showed that OTNA induced NO production through activation of the PI3K/Akt/eNOS pathway in endothelial cells. Furthermore, we also found that OTNA directly bound to AhR and activated the PI3K/Akt/eNOS pathway to dilate pulmonary arteries by inhibiting AhR. Conclusions OTNA relaxes pulmonary arteries by antagonizing AhR. This study provides a new natural antagonist of AhR as a promising lead compound for PAH treatment.

Recent studies have shown that the reduction of sperm count is associated with sleep behaviors, including restricted/excessive sleep duration, late sleep time midpoint, and shift of sleep midpoint time (social jetlag). Chronotype is suggested to regulate sleep behaviors; however, the relationship between chronotype and human sperm count is unknown. The relationship between sleep behaviors and human sperm count, when sleep behaviors as well as chronotype are controlled is also unclear. We performed a path analysis of the data obtained from 667 Chinese men recruited into the MARHCS (Male Reproductive Health in Chongqing College Students) study. Chronotype, sleep duration, sleep time midpoint, and social jetlag were estimated by the Munich Chronotype Questionnaire. Sperm count was measured by computer-aided sperm analysis. The comprehensive relationship between chronotype, all sleep behaviors, and sperm count was tested by path analysis, in which the standardized residual of sperm count was used for adjustment of age, abstinence period, body mass index, tobacco smoking, alcohol drinking, and intake of tea, cola, and coffee. Our major findings are: (1) Chronotype negatively correlated with sleep duration (correlation coefficient (R) = ���0.32, P P P P = .005), sleep midpoint (R = ���0.09, P = .021), and social jetlag (R = ���0.12, P = .001), respectively, correlated with sperm count in univariate analysis. The relationship between sleep midpoint and sperm count disappeared when social jetlag was controlled. (3) Path analysis showed that chronotype is connected with sperm count through two pathways: via sleep duration (standardized path coefficient = ���0.09, 95% CI: ���0.17 to ���0.01, P = .030) and via social jetlag (standardized path coefficient = ���0.09, 95% CI: ���0.17 to ���0.01, P = .031). For men of earlier, intermediate, and later tertile of chronotype, the sleep-duration-related decrease of sperm count was separately estimated to be 16.3%, 12.8%, and 11.6%, while the social-jetlag-related decrease of sperm count was estimated to be 9.3%, 12.8%, and 19.2%. The total effect of sleep behaviors on sperm count was estimated to be 25.7%, 25.6%, and 30.7%, with an average of 27.4% for men of different chronotypes. The present study showed that men of earlier chronotype were prone to restricted/excessive sleep duration, while men of later chronotype were prone to social jetlag, both of which correlated with reduced sperm count, suggesting that chronotype may modulate the sleep behaviors and exert dual effects on sperm count via different sleep behaviors, leading to a ubiquitous sperm decline. Men of different chronotypes should take care to avoid different types of improper sleep behaviors, so as to prevent such deleterious effect on sperm count.

Supplemental Material, sj-pdf-1-ccx-10.1177_10732748211040009 for Association Between LIN28A Gene Polymorphisms and Glioma Susceptibility in Chinese Children by Huiqin Guo, Yuxiang Liao, Ao Lin, Huiran Lin, Xiaokai Huang, Jichen Ruan, Li Yuan and Zhenjian Zhuo in Cancer Control

Air pollution, which is associated with male reproductive health. However, it is unknown the acute effects of ambient air pollutants exposure on male reproductive hormones. The current study, we measured serum levels of reproductive hormone in 2030 blood samples gathered from The Male Reproductive Health in Chongqing College Students (MARHCS) cohort study. We derived a full coverage of ambient air pollutant (PM10, PM2.5, SO2, NO2, CO and O3) concentrations by employing machine learning algorithms, and used a mixed-effect model to estimate single-day and cumulative effects of air pollutants exposure on serum reproductive hormones. Our results showed that (1) PM10 and PM2.5 concentrations were positively associated with estradiol (E2) in both single and cumulative lag days, but were negatively associated with the ratio of Testosterone/E2 (the T/E2 ratio). NO2 was positively associated with estradiol at lag day 2 (95% CI: 0.290, 0.881; corrected P = 0.048) and lag 0–2 days (95% CI: 0.523, 1.337; corrected P = 0.003), with progesterone (P) at lag day 2 and lag day 3 (corrected P

Exposure to outdoor fine particulate matter (PM2.5)-bound polycyclic aromatic hydrocarbons (PAHs) is linked to reproductive dysfunction. However, it is unclear which component of PAHs is responsible for the adverse outcomes. In the Male Reproductive Health in Chongqing College Students (MARHCS) cohort study, we measured the exposure levels of 16 PAHs by collecting air PM2.5 particles and assessed eight PAHs metabolites from four parent PAHs, including naphthalene, fluorene, phenanthrene, and pyrene in urine samples. We investigated compositional profiles and variation characteristics for 16 PAHs in PM2.5, and then assessed the association between PAHs exposure and semen routine parameters, sperm chromatin structure, and serum hormone levels in 1452 samples. The results showed that naphthalene (95% CI: −17.989, −8.101), chrysene (95% CI: −64.894, −47.575), benzo[a]anthracene (95% CI: −63.227, −45.936) and all the high molecular weight (HMW) PAHs in PM2.5 were negatively associated with sperm normal morphology. Most of the low molecular weight (LMW) PAHs, such as acenaphthylene, fluorene, phenanthrene, fluoranthene, pyrene, chrysene, benzo[a]anthracene, ∑LMW PAHs and ∑16 PAHs, were correlated with increased sperm motility (all corrected P

Summary The COVID-19 pandemic caused by SARS-CoV2 is characterized by a remarkable variation in clinical severity ranging from a mild illness to a fatal multi-organ disease. Understanding the dysregulated human immune responses in the fatal subjects is critical for management of COVID-19 patients and the pandemic. In this study, we examined the immune cell compositions in the lung tissues and hilar lymph nodes using immunohistochemistry on 6 deceased COVID-19 patients and 4 focal organizing pneumonia (FOP) patients who underwent lung surgery and served as controls. We found a dominant presence of macrophages and a general deficiency of T cells and B cells in the lung tissues from deceased COVID-19 patients. In contrast to the FOP patients, Tfh cells and germinal center formation were largely absent in the draining hilar lymph nodes in the deceased COVID-19 patients. This was correlated with reduced IgM and IgG levels compared to convalescent COVID-19 patients. In summary, our data highlight a defect of germinal center structure in deceased COVID-19 patients leading to an impaired humoral immunity. Understanding the mechanisms of this deficiency will be one of the key points for the management of this epidemic.

Five novel bisindole alkaloids, hunzeylanines A-E (

Three-dimensional numerical simulation technology was used to investigate the influence of structural and material parameters on the modulation transfer function (MTF) of lattice matched InGaAs/InP planar small pitch arrays. We found that extending depletion zone through increasing depth of diffusion junction and reducing the doping concentration in the absorption layer plays a critical role in optimizing the MTF of arrays. And it is highly worth noting that the very optimization effects become even more pronounced in smaller pitch arrays, which offers valuable references to restrain crosstalk in highly dense array and strengthen the imaging ability of large format photodetector system in future. However, the inevitable cost of deteriorating the specific detectivity should not be ignored. And hence, junction depth and doping concentration of the absorption layer need to be balanced in actual preparation. In addition, the effects of lattice temperature on MTF and crosstalk in arrays were also discussed. The inter-pixel crosstalk deteriorates seriously as rising lattice temperature. A stronger impact of lattice temperature on MTF of larger pitch arrays has been confirmed. This work offers important reference to suppress inter-pixel crosstalk in obtaining the high-resolution imaging for InGaAs photodetector focal plane array.

The protein containing the C2 domain has been well documented for its essential roles in endocytosis, cellular metabolism and cancer. Tac2-N (TC2N) is a tandem C2 domain-containing protein, but its function, including its role in tumorigenesis, remains unknown. Here, we first identified TC2N as a novel oncogene in lung cancer. TC2N was preferentially upregulated in lung cancer tissues compared with adjacent normal lung tissues. High TC2N expression was significantly associated with poor outcome of lung cancer patients. Knockdown of TC2N markedly induces cell apoptosis and cell cycle arrest with repressing proliferation in vitro, and suppresses tumorigenicity in vivo, whereas overexpression of TC2N has the opposite effects both in vitro and in vivo. Using a combination of TCGA database and bioinformatics, we demonstrate that TC2N is involved in regulation of the p53 signaling pathway. Mechanistically, TC2N attenuates p53 signaling pathway through inhibiting Cdk5-induced phosphorylation of p53 via inducing Cdk5 degradation or disrupting the interaction between Cdk5 and p53. Moreover, the blockade of p53 attenuates the function of TC2N knockdown in the regulation of cell proliferation and apoptosis. In addition, downregulated TC2N is involved in the apoptosis of lung cancer cells induced by doxorubicin, leading to p53 pathway activation. Overall, these findings uncover a role for the p53 inactivator TC2N in regulating the proliferation and apoptosis of lung cancer cells. Our present study provides novel insights into the mechanism of tumorigenesis in lung cancer.

Ervadivamines A (1) and B (2), two unprecedented trimeric monoterpenoid indole alkaloids, were isolated from Ervatamia divaricata. They are the first examples of vobasine-iboga-vobasine-type alkaloid with both C-C and C-N linkage patterns. Their structures including absolute configurations were fully accomplished by extensive spectroscopic analysis, single-crystal X-ray diffraction, and electric circular dichroism methods. The plausible biogenetic pathways of these trimeric alkaloids were also proposed. In addition, compound 1 exhibited significant cytotoxicity against four cancer cells.

OBJECTIVE Behavioral and psychosocial factors have been associated with a decline of the quality of semen. However, the relationship of depression and physical activity (PA) with semen quality remains unclear. METHODS Data were obtained from 587 young male Chinese college students in June 2013. Participants completed a questionnaire assessing life-style factors, the Zung self-rated depression scale, and three items related to PA. They underwent a physical examination and provided a semen sample and a blood sample for reproductive hormones (testosterone, estrogen, progesterone, follicle-stimulating hormone, luteinizing hormone, and prolactin). RESULTS Men with high depression scores (n = 63, 10.7%) had lower sperm concentration (M (SD) = 66.9 (74.5) versus 72.6 (56.9) [10/ml], p = .043) and total sperm count (M (SD) = 241.6 (299.7) versus 257.0 (204.0) [10], p = .024) than nondepressed men. Participants with low PA levels (n = 99, 16.9%) had lower total sperm count (M (SD) = 204.4 (153.7) versus 265.8 (225.8) [10/ml], p = .017) than participants with higher activity levels. After adjusting for potential confounders, depressed men had 18.90% (95% confidence interval [CI] = 1.14%-33.47%) lower sperm concentration and 21.84% (95% CI = 3.39%-36.90%) lower total sperm count than nondepressed men. Men with low PA levels had 23.03% (95% CI = 2.80%-46.89%) lower total sperm count than physically active participants. An interaction effect between depression and PA on sperm concentration was detected (p = .033). There were no significant associations of depression and PA with reproductive hormones (p > .05). CONCLUSIONS Depression and low levels of PA are associated with lower levels of semen quality, which may have implications for reproductive health.

Sertoli cells (SCs) provide physical and nutritional support for spermatogenesis. Dibutyl phthalate (DBP) is a plasticizer that has male reproductive toxicity. The collapse of vimentin in DBP-exposed SCs is thought to induce the sloughing of spermatocytes from seminiferous tubules. In this study, we explored methods to rescue vimentin from collapse in DBP-exposed SCs. DBP not only induced the hyperphosphorylation of vimentin but also triggered endoplasmic reticulum (ER) stress and apoptosis in SCs. Treatment with BAPTA-AM, an antagonist of Ca2+, significantly decreased the level of phosphorylated vimentin, while LY294002, an inhibitor of Akt1, did not. ER stress and apoptosis remained at high levels, and the distribution of vimentin was not improved. ZnSO4 treatment did not decrease the level of phosphorylated vimentin. However, after treatment, ER stress and apoptosis were obviously inhibited, and the distribution of vimentin was reconverted. These results indicated that ZnSO4 could alleviate the collapse of vimentin by attenuating ER stress and apoptosis. This study suggested that an appropriate zinc supply might be a choice to alleviate DBP-induced adverse reproductive effects.

To study the associations between pubertal timing and semen quality and reproductive hormones, 680 volunteers were recruited from universities in Chongqing, China. Pubertal timing was obtained using a questionnaire. The main measurements were five routine semen parameters and six reproductive hormones. After adjusting for potential confounders, we found that early pubertal timing was associated with lower sperm concentration. An one-year increase in age of peak height velocity was associated with a 4.7% (95% confidence interval [CI] = 1.0 to 8.6) increase in sperm concentration. An one-year increase in age of first spermatorrhea was associated with a 6.4% increase in sperm concentration and a 2.9% decrease in semen volume (95% CI = 1.7 to 11.3, −5.5 to −0.3; respectively). Regarding reproductive hormones, an one-year increase in age of height spurt and peak height velocity was associated with a 6.5% and a 6.7% decrease in estrogen (95% CI = −9.8 to −3.0, −10.4 to −2.8; respectively). While an one-year increase in age of height spurt was associated with higher follicle-stimulating hormone (% change = 2.6, 95% CI = 0.2 to 4.7). This was the first report that has suggested that early pubertal timing is associated with lower sperm concentration. However, further study is still needed to validate this association and fully elucidate the mechanism behind it.