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Background: Endoplasmic reticulum (ER) stress, with adaptive unfolded protein response (UPR), is a key link between obesity, insulin resistance and type 2 diabetes, all of which are often present in the most common endocrine-metabolic disorder in women of reproductive age, polycystic ovary syndrome (PCOS), which is characterized with hyperandrogenism. However, the link between excess androgen and Endoplasmic reticulum (ER) stress/insulin resistance in patients with polycystic ovary syndrome (PCOS) is unknown. Methods: An unexpected role of kisspeptin was reported in the regulation of UPR pathways and its involvement in the androgen-induced ER stress in hypothalamic neuronal cells. To evaluate the relationship of kisspeptin and ER stress, We detected Kisspeptin and other factors in blood plasm of PCOS patients, rat models and hypothalamic neuronal cells. Findings: We detected higher testosterone and lower kisspeptin levels in the plasma of PCOS than that in non-PCOS women. We established a PCOS rat model by dihydrotestosterone (DHT) chronic exposure, and observed significantly down-regulated kisspeptin expression and activated UPR pathways in PCOS rat hypothalamus compared to that in controls. Inhibition or knockdown of kisspeptin completely mimicked the enhancing effect of DHT on UPR pathways in a hypothalamic neuronal cell line, GT1-7. Kp10, the most potent peptide of kisspeptin, effectively reversed or suppressed the activated UPR pathways induced by DHT or thapsigargin, an ER stress activator, in GT1-7 cells, as well as in the hypothalamus in PCOS rats. Similarly, Kisspeptin attenuated thapsigargin-induced Ca2+ response and the DHT- induced insulin resistance in GT1-7 cells. Interpretation: Our data has revealed an unexpected protective role of kisspeptin against ER stress and insulin resistance in the hypothalamus and provided a new treatment strategy targeting hypothalamic ER stress and insulin resistance with kisspeptin as a potential therapeutic agent. Funding Statement: National Basic Research Program of China, National Natural Science Foundation of China (2012CB944902, 2012CB944703, 2013CB967404, 2013CB967401, 2013CB967403, 81571403, 81471429) and State Key laboratory of Reproductive Medicine (SKLRM-K201805). Declaration of Interests: The authors have no conflicts of interest about this work. Ethics Approval Statement: The study was approved by the Ethics Committee of Tongji University and all patients gave informed consent. All procedures in this study were approved by the Animal Experimentation Ethics Committee of the Chinese University of Hong Kong, and the methods were used in accordance with the guidelines of the Chinses University of Hong Kong.

The nickel isotope has a great potential for investigating biogeochemical processes in the geosciences and has been proven to be extremely useful for studying the chronology of the early solar system. As one of the most powerful techniques in metal isotopic ratio measurements, thermal ionization mass spectrometry (TIMS) shows poor sensitivity for Ni isotope ratio analysis, because the high ionization potential (7.640eV) of Ni makes it difficult to be ionized on a hot filament. In previous studies, a minimum of 1000 ng Ni was required using the classical Sigel-H3BO3-Al emitter or the Sigel-H3PO4-Al emitter. The present study is first to employ a highly sensitive zirconium hydrogen phosphate (Zr(HPO4)2) emitter to measure the Ni isotopes with TIMS. This emitter produces a significant enhancement in the ionization efficiency of Ni and enables the analysis of the Ni isotope at the 200 ng level with high precision. In addition, this newly developed emitter shows a strong inhibition to the Fe and Zn signals, which are the main potential isobaric interferences during Ni measurement. A series of analyses of the NIST SRM 986 standard demonstrated that an internal precision (2 RSE) of ± 0.03 to 0.05 ‰ can be achieved depending on the sample size. Analysis of the NIST SRM 986 standard also shows an external reproducibility (2 RSD, n = 10) of better than ± 0.11‰, even for Ni at 200 ng.

Thermal ionization mass spectrometry (TIMS) has shown excellent analytical precision for Sr isotopic ratio analysis, even for small masses of material (0.5-10 ng). However, because of the sensitivity limit of TIMS, it is still not possible to obtain high precision Sr/Sr isotope ratios for picogram-level sample sizes (30-100 pg) due to the lack of a highly sensitive emitter. This study is the first to employ a highly sensitive silicotungstic acid emitter to measure Sr isotopes at the picogram-level using TIMS. This emitter produces a 3-fold enhancement in the ionization efficiency of Sr and not only significantly reduces the required sample size but also has good external precision. Analyses of the NIST 987 standard yield an external reproducibility (2 RSD, n = 8) better than ±0.013% even for 30 pg of Sr. It is possible to yield an internal precision (2 RSE) of ±0.003% for 100 pg of sample using the default 10 Ohm feedback resistors. This method was verified by using a suite of silicate reference materials. Replicate digestions and analyses (n = 8) of the basalt standard BCR-2 (Sr/Sr = 0.704998 ± 0.000028, 2 SD) at the 326 ± 30 pg level demonstrates that good external reproducibility is reached on ultratrace level silicate samples. This method has a wide variety of potential applications for samples containing ultralow amounts of Sr in geoscience and archeological studies, such as single grains of mica, sphalerite, and pyrite, single mantle melt inclusions, precious extra-terrestrial materials, and human hair to name just a few.

Patients with inflammation in the terminal ileum have high morbidity. In genetically susceptible hosts, chronic intestinal inflammation targeting the resident intestinal microbiota develops, but the microbial signature of the terminal ileum is poorly studied. To improve understanding of the mechanisms underlying the high prevalence of terminal ileum inflammation, we used 16S rRNA sequencing to analyse the mucosa-associated microbiota of the terminal ileum under intestinal homeostasis and inflammation conditions. Mucosal biopsy is the most commonly used sampling technique for assessing microbial communities associated with the intestinal mucosa. Thirty patients (15 with terminal ileum inflammation and 15 controls) underwent colonoscopy and biopsies were taken from the terminal ileum. Diagnosis depended on a combination of endoscopic and histological factors. To determine the composition and diversity of the microbiota, the 16S rRNA was analysed, and a variety of bioinformatics analyses were performed. Among the patients, composition analysis showed that the most abundant phyla identified in the terminal ileum samples were Fusobacteria, Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria. At the phylum level, the relative proportion of Bacteroidetes was lower in patients with inflammation than in control patients. In addition, there was an increase in the abundance of the phyla Proteobacteria and Lentisphaerae in patients with inflammation. The abundances of the dominant microbes in the terminal ileum were not significantly different between patients in an inflammatory state and controls. These results confirm that partial dysbiosis of the intestinal mucosa-associated microbiota composition is associated with terminal ileum inflammation.

The shift of cytokine profile from anti‐ to pro‐inflammatory is the most recognizable sign of labor, although the underlying mechanism remains elusive. Here, we report that the epithelial sodium channel (ENaC) is upregulated and activated in the uterus at labor in mice. Mechanical activation of ENaC results in phosphorylation of CREB and upregulation of pro‐inflammatory cytokines as well as COX‐2/PGE 2 in uterine epithelial cells. ENaC expression is also upregulated in mice with RU486‐induced preterm labor as well as in women with preterm labor. Interference with ENaC attenuates mechanically stimulated uterine contractions and significantly delays the RU486‐induced preterm labor in mice. Analysis of a human transcriptome database for maternal–fetus tissue/blood collected at onset of human term and preterm births reveals significant and positive correlation of ENaC with labor‐associated pro‐inflammatory factors in labored birth groups (both term and preterm), but not in non‐labored birth groups. Taken together, the present finding reveals a pro‐inflammatory role of ENaC in labor at term and preterm, suggesting it as a potential target for the prevention and treatment of preterm labor.

Chromium isotopes have important applications in cosmochemistry, environmental science, geochemistry, and paleoceanography studies. In previous investigations, thermal ionization mass spectrometry (TIMS) showed excellent precision for Cr isotopic ratios analysis. However, relative large sample sizes (200–2000 ng) were required for TIMS measurements and therefore hampered its wider application in geoscience and environmental science. In this study, a highly sensitive Nb2O5 emitter is employed for the first time to measure Cr isotopes using TIMS. This Nb2O5 emitter produces at least a 10-fold enhancement in the sensitivity and can not only significantly reduce the sample size, but also obtain good external precision. In this study, an external precision of ±0.23‰ (2 SD, n = 10) on δ53Cr was obtained on a 5 ng sample of NIST SRM 3112a standard. In addition, an improved two-column chemical separation procedure was devised in order to separate Cr from complex geological materials. Compared to the classical anion resin purification method, the procedural blank and severe inhibition effect from SO42− is significantly reduced by carefully optimizing and evaluating the dosage of (NH4)2S2O8 oxidant. This method has been verified using one dolomite, three basalt and one peridotite reference materials. A range of analytical tests demonstrates that it is possible to attain internal precisions (2 SE) of ±0.017 to 0.037‰ on a single 30 min analysis. Long-term reproducibility (2 SD) for 53Cr/52Cr is 0.113454 ± 0.000004 (∼0.035‰) obtained by replicate analyses (n = 105) of NIST SRM 3112a over a 6 month period. Replicate digestions and analyses of the basalt standard BIR-1 (δ53Cr = −0.110 ± 0.048‰, 2 SD) and the carbonate standard JDo-1 (δ53Cr = 1.710 ± 0.054‰, 2 SD) demonstrate that good external reproducibility is obtainable on natural samples.

A straightforward tandem column separation procedure is presented for the separation of Sr, Nd, Pb, and Hf from silicate materials. It allows rapid purification, without any intervening evaporation, of these four elements of great interest in Earth science and cosmochemistry. After sample loading, the upper Sr Spec column adsorbs Sr and Pb, while the lower TODGA Spec column adsorbs Hf and Nd. Strontium-lead and hafnium–neodymium elements are then back-extracted from the Sr Spec and TODGA Spec columns, respectively. The whole separation procedure, including column setup, cleaning, and pre-conditioning, takes approximately eight hours for separating a batch of 25 samples. The proposed procedure offers significant improvement in separation efficiency of these often-used four elements, compared with conventional four column methods. Fractions of Sr, Nd and Pb are then measured by TIMS and the Hf fraction is determined by MC-ICP-MS. The stability of this procedure was demonstrated by replicate measurements of 87Sr/86Sr, 143Nd/144Nd, 176Hf/177Hf, 206Pb/204Pb, 207Pb/204Pb and 208Pb/204Pb isotope ratios of eight international silicate rock reference materials, spanning a wide range of bulk compositions. The analytical results obtained in this study agree well with published data. The external reproducibility (2RSD, n = 8) of standard BCR-2 was ±0.0026% for 87Sr/86Sr, ±0.0020% for 143Nd/144Nd, ±0.0049% for 176Hf/177Hf, and ±0.026–0.034% for 206Pb/204Pb, 207Pb/204Pb and 208Pb/204Pb isotope ratios.

Polycystic ovarian syndrome (PCOS) is one of the most frequent causes of female infertility, featured by abnormal hormone profile, chronic oligo/anovulation, and presence of multiple cystic follicles in the ovary. However, the mechanism underlying the abnormal folliculogenesis remains obscure. We have previously demonstrated that CFTR, a cAMP-dependent Cl−and HCO3−conducting anion channel, is expressed in the granulosa cells and its expression is downregulated in PCOS rat models and human patients. In this study, we aimed to investigate the possible involvement of downregulation of CFTR in the impaired follicle development in PCOS using two rat PCOS models and primary culture of granulosa cells. Our results indicated that the downregulation of CFTR in the cystic follicles was accompanied by reduced expression of proliferating cell nuclear antigen (PCNA), in rat PCOS models. In addition, knockdown or inhibition of CFTR in granulosa cell culture resulted in reduced cell viability and downregulation of PCNA. We further demonstrated that CFTR regulated both basal and FSH-stimulated granulosa cell proliferation through the HCO3−/sAC/PKA pathway leading to ERK phosphorylation and its downstream target cyclin D2(Ccnd2) upregulation. Reduced ERK phosphorylation and CCND2 were found in ovaries of rat PCOS model compared with the control. This study suggests that CFTR is required for normal follicle development and that its downregulation in PCOS may inhibit granulosa cell proliferation, resulting in abnormal follicle development in PCOS.

Thermal ionization mass spectrometry (TIMS) is considered the most accurate technique for determining Sr–Nd–Pb isotopic ratios in geological samples. However, time-consuming and complex sample separation procedures greatly hinder the instrumental measurement efficiency. In this study, a single-column separation chemistry procedure for Sr–Nd–Pb from single rock dissolution was developed. The chemistry procedure was designed to minimize the number of evaporation steps and considerably shorten the separation time, enabling high throughput for TIMS. In contrast to conventional three-column separation procedures (∼3 days), this technique was characterized by high efficiency superiority in terms of separation time (∼8 hours), a 3-fold enhancement in the separation efficiency. The stability of our procedure was demonstrated by replicated TIMS measurements of 87Sr/86Sr, 143Nd/144Nd, 206Pb/204Pb, 207Pb/204Pb and 208Pb/204Pb ratios for six international silicate rock reference materials, spanning a wide range of bulk compositions. The analytical results obtained for these standards agreed well with published data. The external reproducibility (2 RSD, n = 10) of a BCR-2 standard sample was ±0.0020% for 87Sr/86Sr, ±0.0023% for 143Nd/144Nd, and ±0.021–0.033% for 206Pb/204Pb, 207Pb/204Pb and 208Pb/204Pb ratios.

Glucagon, produced by islet α cells, functions to increase blood glucose. Abnormal glucose levels are often seen in cystic fibrosis (CF), a systematic disease caused by mutations of the CF transmembrane conductance regulator (CFTR), and in polycystic ovarian syndrome (PCOS), an endocrine disorder featured with hyperandrogenism affecting 5–10% women of reproductive age. Here, we explored the role of CFTR in glucagon production in α cells and its possible contribution to glucagon disturbance in CF and PCOS. We found elevated fasting glucagon levels in CFTR mutant (DF508) mice compared to the wildtypes. Glucagon and prohormone convertase 2 (PC2) were also upregulated in CFTR inhibitor-treated or DF508 islets, as compared to the controls or wildtypes, respectively. Dihydrotestosterone (DHT)-induced PCOS rats exhibited significantly lower fasting glucagon levels with higher CFTR expression in α cells compared to that of controls. Treatment of mouse islets or αTC1-9 cells with DHT enhanced CFTR expression and reduced the levels of glucagon and PC2. The inhibitory effect of DHT on glucagon production was blocked by CFTR inhibitors in mouse islets, and mimicked by overexpressing CFTR in αTC1-9 cells with reduced phosphorylation of the cAMP/Ca2+ response element binding protein (p-CREB), a key transcription factor for glucagon and PC2. These results revealed a previously undefined role of CFTR in suppressing glucagon production in α-cells, defects in which may contribute to glucose metabolic disorder seen in CF and PCOS.

The secretion of glucagon by islet α cells is normally suppressed by high blood glucose, but this suppressibility is impaired in patients with diabetes or cystic fibrosis (CF), a disease caused by mutations in the gene encoding CF transmembrane conductance regulator (CFTR), a cyclic adenosine monophosphate-activated Cl- channel. However, precisely how glucose regulates glucagon release remains controversial. Here we report that elevated glucagon secretion, together with increased glucose-induced membrane depolarization and Ca2+ response, is found in CFTR mutant (DF508) mice/islets compared with the wild-type. Overexpression of CFTR in AlphaTC1-9 cells results in membrane hyperpolarization and reduced glucagon release, which can be reversed by CFTR inhibition. CFTR is found to potentiate the adenosine triphosphate-sensitive K+ (KATP) channel because membrane depolarization and whole-cell currents sensitive to KATP blockers are significantly greater in wild-type/CFTR-overexpressed α cells compared with that in DF508/non-overexpressed cells. KATP knockdown also reverses the suppressive effect of CFTR overexpression on glucagon secretion. The results reveal that by potentiating KATP channels, CFTR acts as a glucose-sensing negative regulator of glucagon secretion in α cells, a defect of which may contribute to glucose intolerance in CF and other types of diabetes.

The increase in migratory ability of pancreatic ductal adenocarcinoma cells is a key event in the development of metastasis to the lymph nodes and distant organs. Although the C-C motif chemokine receptor 7 (CCR7) and its ligand, C-C motif chemokine ligand 21 (CCL21), have been revealed to serve an important role in tumor migration, their precise roles and potential underlying mechanisms remain largely unknown. The present study revealed that overexpression of CCR7 significantly promoted BxPC-3 cell migration, accompanied by the induction of anoctamin 6 (ANO6) expression, indicating that ANO6 is a downstream target of CCR7 signaling. Furthermore, the level of phosphorylated extracellular signal-regulated kinase (ERK) was significantly increased in CCR7-overexpressing BxPC-3 cells, indicating that ERK may be a potential mediator of CCR7-regulated ANO6 expression in BxPC-3 cells. To characterize the receptor-mediated pathway, a specific ERK inhibitor, U0126, was used, which reduced BxPC-3 cell migration and the expression of ANO6. In summary, the results of the present study demonstrate that CCR7 promoted BxPC-3 cell migration by regulating ANO6 expression perhaps via activation of the ERK signaling pathway.

Ethnopharmacological relevance Disturbed gastrointestinal (GI) motility can be associated with smooth muscle abnormalities and dysfunction. Exploring innovative approaches that can modulate the disturbed colonic motility are of great importance for clinical therapeutics. Naringenin, a flavonoid presented in many traditional Chinese herbal medicines, has been shown to have a relaxant effect on different smooth muscles. The aim of the present study was to investigate the effect of naringenin on regulation of GI motility. Material and methods Mechanical recording was used to investigate the effect of naringenin on isolated rat colonic smooth muscle spontaneous contractions. Whole cell patch clamp, intracellular [Ca2+] concentration ([Ca2+]i) and membrane potential measurements were examined on primary cultures of colonic smooth muscle cells (SMCs). A neostigmine-stimulated rat model was utilized to investigate the effect of naringenin in vivo. Results Naringenin induced a concentration-dependent inhibition (1–1000 μM) on rat colonic spontaneous contraction, which was reversible after wash out. The external Ca2+ influx induced contraction and [Ca2+]i increase were inhibited by naringenin (100 μM). In rat colonic SMCs, naringenin-induced membrane potential hyperpolarization was sensitive to TEA and selective large-conductance calcium-activated K+ (BKCa) channel inhibitor iberiotoxin. Under whole cell patch-clamp condition, naringenin stimulated an iberiotoxin-sensitive BKCa current, which was insensitive to changes in the [Ca2+]i concentration. Furthermore, naringenin significantly suppressed neostigmine-enhanced rat colon transit in vivo. Conclusion Our results for the first time demonstrated the relaxant effect of flavonoid naringenin on colon smooth muscle both in vitro and in vivo. The relaxant effect of naringenin was attributed to direct activation of BKCa channels, which subsequently hyperpolarized the colonic SMCs and decreased Ca2+ influx through VDCC. Naringenin might be of therapeutic value in the treatment of GI motility disorders.

Thermal ionization mass spectrometry and multiple-collector inductively coupled plasma mass spectrometry are considered to be “gold standards” for the determination of the isotope ratios of Sr–Nd and Hf in geological samples because of the extremely high precision and accuracy of these methods. However, the sample throughputs are hindered by time-consuming and tedious chemical procedures. Three-step ion exchange resin separation is traditionally employed to purify Sr–Nd–Hf from matrix elements. In this study, a one-step Sr–Nd–Hf separation scheme was developed to process geological samples. The separation scheme is based on the combined use of conventional AG50W-X12 cation-exchange resin and LN Spec extraction chromatographic material without any intervening evaporation step. The protocol not only prevents cross-contamination during operation using multiple-stage ion exchange resins but also significantly improves the efficiency of sample preparation. The stability of our chemical procedure was demonstrated by replicate measurements of 87Sr/86Sr, 143Nd/144Nd, and 176Hf/177Hf ratios in six international reference materials of silicate rocks. The analytical results obtained for these standard rocks compare well with the published data. The external reproducibility (2 SD, n = 10) of a BCR-2 standard sample was ±0.000018 for 87Sr/86Sr, ±0.000010 for 143Nd/144Nd, and ±0.000014 for 176Hf/177Hf.

Estrogens play important roles in a wide range of physiological and pathological processes, and their biosynthesis is profoundly influenced by FSH that regulates the rate-limiting enzyme aromatase-converting estrogens from androgens. Abnormal estrogen levels are often seen in diseases such as ovarian disorders in polycystic ovarian syndrome (PCOS), an endocrine disorder affecting 5-10% of women of reproductive age, and cystic fibrosis (CF), a common genetic disease caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR).We undertook the present study to investigate the mechanism underlying these ovarian disorders, which is not well understood.FSH-stimulated cAMP-responsive element binding protein phosphorylation, aromatase expression, and estradiol production are found to be enhanced by HCO3- and a HCO3- sensor, the soluble adenylyl cyclase, which could be significantly reduced by CFTR inhibition or in ovaries or granulosa cells of cftr knockout/ΔF508 mutant mice. CFTR expression is found positively correlated with aromatase expression in human granulosa cells, supporting its role in regulating estrogen production in humans. Reduced CFTR and aromatase expression is also found in PCOS rodent models and human patients.CFTR regulates ovarian estrogen biosynthesis by amplifying the FSH-stimulated signal via the nuclear soluble adenylyl cyclase. The present findings suggest that defective CFTR-dependent regulation of estrogen production may underlie the ovarian disorders seen in CF and PCOS.

Background The cystic fibrosis transmembrane conductance regulator (CFTR) plays a critical role in electrolyte and fluid transport in epithelial cells, and women with cystic fibrosis (CF), caused by CFTR gene mutations, have a higher incidence of infertility. Methods In the present study, we investigated the expression of CFTR in porcine oviduct and its functional role in oviductal HCO(3)(-) secretion and embryo development with RT-PCR, western blot, patch-clamp, short-circuit current (I(sc)), pH measurement and embryo culture. Results RT-PCR and western blot analysis showed the expression of CFTR mRNA and protein in the oviduct with its localization demonstrated by immunohistochemistry. The whole-cell patch-clamp recording revealed a forskolin (FSK)-activated current with electrophysiological and pharmacological characteristics of CFTR. The I(sc) measurement showed that FSK-stimulated an increase in the I(sc), which could be significantly reduced by CFTR inhibitor or removal of both CO(2) and HCO(3)(-). pH measurement showed a FSK stimulated alkalization at the apical surface, which could be inhibited by CFTR inhibitor, indicating CFTR-mediated HCO(3)(-) secretion. Mouse embryo development from 2-cell to morula or blastocyst stage was significantly inhibited in the absence of HCO(3)(-) or when co-cultured with HCO(3)(-) secretion-deficient CFTR mutant cells as compared with the wild-type. RT-PCR, western blot and immunostaining showed the expression of soluble adenylate cyclase (sAC), the known HCO(3)(-) sensor, in embryos. Treatment with its inhibitors, 2-hydroxyestradiol and KH7, prevented the HCO(3)(-) dependent embryo development. Conclusion The present results suggest that CFTR-mediated oviductal HCO(3)(-) secretion may be vital for sAC-dependent early embryo development, a defect of which may contribute to the reduced fertility seen in women with CF.

Recent studies suggest that the epithelium might modulate the contractility of smooth muscle. However, the mechanisms underlying this regulation are unknown. The present study investigated the regulation of smooth muscle contraction by the epithelium in rat vas deferens and the possible factor(s) involved. Exogenously applied ATP inhibited electrical field stimulation (EFS)-evoked smooth muscle contraction in an epithelium-dependent manner. As theeffectsofATPonsmoothmusclecontractilitywereabrogatedbyinhibitorsofprostaglandin synthesis, but not by those of nitric oxide synthesis, prostaglandins might mediate the effects of ATP. Consistent with this idea, PGE2 inhibited EFS-evoked smooth muscle contraction independent of the epithelium, while ATP and UTP induced the release of PGE2 from cultured rat vas deferens epithelial cells, but not smooth muscle cells. The ATP-induced PGE2 release from vas deferens epithelial cells was abolished by U73122, an inhibitor of phospholipase C (PLC) and BAPTAAM, a Ca 2+ chelator. ATP also transiently increased [Ca 2+ ]i in vas deferens epithelialcells.ThiseffectofATPon[Ca 2+ ]iwasindependentofextracellularCa 2+ ,butabolished by the P2 receptor antagonist RB2 and U73122. In membrane potential measurements using a voltage-sensitive dye, PGE2, but not ATP, hyperpolarized vas deferens smooth muscle cells and this effect of PGE2 was blocked by MDL12330A, an adenylate cyclase inhibitor, and the chromanol293B,ablockerofcAMP-dependentK + channels.Takentogether,ourresultssuggest that ATP inhibition of vas deferens smooth muscle contraction is epithelium dependent. The data also suggest that ATP activates P2Y receptor-coupled Ca 2+ mobilization leading to the release of PGE2 from epithelial cells, which in turn activates cAMP-dependent K + channels in smooth muscle cells leading to the hyperpolarization of membrane voltage and the inhibition of vas deferens contraction. Thus, the present findings suggest a novel regulatory mechanism by which the epithelium regulates the contractility of smooth muscle.

Supplemental Digital Content is available in the text, Human adenovirus 36 (HAdV-36), as the key pathogen, was supposed and discussed to be associated with obesity. We searched the references on the association between HAdV-36 infection and obesity with the different epidemiological methods, to explore the relationship with a larger sample size by meta-analysis and compare the differences of epidemiological methods and population subsets by the subgroup analyses. We conducted literature search on the association between HAdV-36 infections and obesity in English or Chinese published up to July 1, 2015. The primary outcome was the HAdV-36 infection rate in the obese and lean groups; the secondary outcomes were the BMI level and BMI z-score in the HAdV-36 positive and negative groups. The pooled odds ratio (OR) was calculated for the primary outcome; the standardized mean differences (SMDs) were calculated for the secondary and third outcomes. Prediction interval (PI) was graphically presented in the forest plot of the random effect meta-analyses. Metaregression analysis and subgroup analysis were performed. Finally 24 references with 10,191 study subjects were included in the meta-analysis. The obesity subjects were more likely to be infected with HAdV-36 compared to the lean controls (OR = 2.00; 95%CI: 1.46, 2.74; PI: 0.59, 6.76; P

The North China Craton (NCC) is proposed to have formed by collision between the Western and Eastern blocks at ∼1.85 Ga. However, the detailed history of the individual blocks remains unsolved. In the Eastern block, the Paleoproterozoic volcano-sedimentary rocks are mainly exposed along its eastern margin, and are referred to as the Liaohe Group in the Liaodong Peninsula of Liaoning Province and as the Guanghua, Ji’an and Laoling Groups in the Tonghua area of southern Jilin Province. The Guanghua and Ji’an Groups are characterized by the occurrence of volcanic rocks, whereas the Laoling Group is composed of clastic sediments and carbonates. Zircon U–Pb isotopic dating by LA-ICP-MS and SHRIMP methods indicates that the Guanghua and Ji’an Groups were formed between 2.1 and 2.0 Ga, but that the rocks of the Laoling Group may have formed in two episodes at 2.5–2.2 and 2.0–1.9 Ga, respectively. The present data indicate that the Ji’an, Laoling and Liaohe Groups are chaotic, without a common stratigraphy, with intense deformation making it difficult to identify original layering and contact relationships. Combined with age data from rocks intrusive into the sequence, the Paleoproterozoic tectonic evolution of the Liaoning–Jilin area can be related to collision and extension between the Liaonan block to the south and the Longgang block to the north. Extension in the Longgang block occurred at 2.12–1.98 Ga and was represented by eruption of the Guanghua mafic volcanic rocks. However, extension in the Liaonan block is expressed by 2.16–2.09 Ga A-type granites and ∼2.12–1.98 Ga volcano-sedimentary rocks of the Ji’an Group. These two blocks collided at ∼1.93–1.90 Ga, resulting in a clockwise metamorphic P – T path recorded in the N. Liaohe–Laoling Groups, followed by ∼1.85 Ga post-collisional evolution expressed by anticlockwise metamorphic P – T paths and immense granitic intrusions in the S. Liaohe–Ji’an Groups. Therefore, it is suggested that the collision between the Longgang and Liaonan in the Eastern block of the NCC occurred a little earlier than ∼1.85 Ga, when the NCC amalgamated by collision of the Western and Eastern blocks.

In our previous study, we have demonstrated that the epithelial sodium channel (ENaC) mediates the embryo-derived signals leading to the activation of CREB and upregulation of cyclooxygenase type 2 (COX2) required for embryo implantation. This study aims to investigate whether microRNAs (miRNAs) are involved in the ENaC-induced upregulation of COX2 during embryo implantation. The results show that the levels of miR-101 and miR-199a-3p, two COX2 targeting miRNAs, are reduced by ENaC activation, and increased by ENaC inhibition or knock-down of ENaC subunit (ENaCα) in human endometrial surface epithelial (HES) cells or in mouse uteri during implantation. Phosphorylation of CREB is induced by the activation of ENaC, and blocked by ENaC inhibition or knockdown in HES cells. Knockdown of ENaCα or CREB in HES cells or in mouse uterusin vivoresults in increases in miR-101 and miR-199a-3p, accompanied with decreases in COX2 protein levels and reduction in implantation rate. The downregulation of COX2 caused by knockdown of ENaC or CREB can be recovered by the inhibitors of miR-101 or miR-199a-3p in HES cells. These results reveal a novel molecular mechanism modulating COX2 expression during embryo implantation via ENaC-dependent CREB activation and COX2-targeting miRNAs.

The cause of insulin insufficiency remains unknown in many diabetic cases. Up to 50% adult patients with cystic fibrosis (CF), a disease caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR), develop CF-related diabetes (CFRD) with most patients exhibiting insulin insufficiency. Here we show that CFTR is a regulator of glucose-dependent electrical acitivities and insulin secretion in β-cells. We demonstrate that glucose elicited whole-cell currents, membrane depolarization, electrical bursts or action potentials, Ca2+ oscillations and insulin secretion are abolished or reduced by inhibitors or knockdown of CFTR in primary mouse β-cells or RINm5F β-cell line, or significantly attenuated in CFTR mutant (DF508) mice compared with wild-type mice. VX-809, a newly discovered corrector of DF508 mutation, successfully rescues the defects in DF508 β-cells. Our results reveal a role of CFTR in glucose-induced electrical activities and insulin secretion in β-cells, shed light on the pathogenesis of CFRD and possibly other idiopathic diabetes, and present a potential treatment strategy.

a b s t r a c t Thermal ionization mass spectrometry (TIMS) offers the excellent precision and accuracy of the Sr and Nd isotopic ratio analysis for geological samples, but this method is labour intensive, expensive and time-consuming. In this study, a new analytical protocol by TIMS is presented that aims at improving analytical efficiency and cutting down experimental cost. Using the single-step cation exchange resin technique, mixed Sr and rare earth elements (REEs) fractions were separated from matrix and evaporated to dryness. Afterwards, mixed Sr + REEs fractions were dissolved and loaded onto the same Re filament using 1 L of 2 M HCl. Then, Sr and Nd were sequentially measured without venting using TIMS. In contrast to conventional TIMS methods, the merits of this analytical protocol are its cost- and time- saving adaptations. The applicability of our method is evaluated by replicated measurements of 87Sr/86Sr and 143 Nd/ 144 Nd for nine international silicate rock reference materials, spanning a wide range of bulk compositions. The typical internal precision in this study is ca. 0.001% (RSE) for 87Sr/86Sr and 143Nd/144Nd; the analytical results obtained for these standard rocks show a good agreement with reported values, indicating the effectiveness of the proposed method.

Cystic fibrosis (CF) is an autosomal recessive disorder caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-dependent anion channel mostly expressed in epithelia. Accumulating evidence suggests that CF airway epithelia are overwhelmed by excessive inflammatory cytokines and prostaglandins (PGs), which eventually lead to the over-inflammatory condition observed in CF lung disease. However, the exact underlying mechanism remains elusive. In this study, we observed increased cyclooxygenase-2 (COX-2) expression and over-production of prostaglandin E(2) (PGE(2)) in human CF bronchial epithelia cell line (CFBE41o--) with elevated NF-κB activity compared to a wild-type airway epithelial cell line (16HBE14o--). Moreover, we demonstrated that CFTR knockout mice had inherently higher levels of COX-2 and NF-κB activity, supporting the notion that lack of CFTR results in hyper-inflammatory signaling. In addition, we identified a positive feedback loop for production of PGE(2) involving PKA and transcription factor, CREB. More importantly, overexpression of wild-type CFTR significantly suppressed COX-2 expression in CFBE41o- cells, and wild-type CFTR protein expression was significantly increased when 16HBE14o-- cells were challenged with LPS as well as PGE(2), indicating possible involvement of CFTR in negative regulation of COX-2/PGE(2). In conclusion, CFTR is a negative regulator of PGE(2)-mediated inflammatory response, defect of which may result in excessive activation of NF-κB, leading to over production of PGE(2) as seen in inflammatory CF tissues.

Embryo implantation remains a poorly understood process. We demonstrate here that activation of the epithelial Na⁺ channel (ENaC) in mouse endometrial epithelial cells by an embryo-released serine protease, trypsin, triggers Ca²⁺ influx that leads to prostaglandin E₂ (PGE₂) release, phosphorylation of the transcription factor CREB and upregulation of cyclooxygenase 2, the enzyme required for prostaglandin production and implantation. We detected maximum ENaC activation, as indicated by ENaC cleavage, at the time of implantation in mice. Blocking or knocking down uterine ENaC in mice resulted in implantation failure. Furthermore, we found that uterine ENaC expression before in vitro fertilization (IVF) treatment is markedly lower in women with implantation failure as compared to those with successful pregnancy. These results indicate a previously undefined role of ENaC in regulating the PGE₂ production and release required for embryo implantation, defects that may be a cause of miscarriage and low success rates in IVF.

To establish decision tree and logistic regression classification models for diagnosing pancreatic adenocarcinoma (PaCa) and for screening serum biomarkers related to evaluation of different stages and curative effects.Serum samples obtained from subjects with pancreatic adenocarcinoma (n = 58) and normal pancreas (n = 51) were applied to strong anion exchange chromatography (SAX2) chips for protein profiling by SELDI-TOF-MS to screen multiple serum biomarkers. Biomarker Wizard software and several statistical methods including algorithm of decision tree, logistic regression and ROC curves were used to construct the decision tree or logistic regression classification models.Average of 61 mass peaks were detected at the molecular range of 2000-30,000, ten decision trees with the highest cross validation rate were chosen to construct the classification models, which can differentiate PaCa from normal pancreas with a sensitivity of 83.3% and a specificity of 100%. Logistic regression was used to achieve the AUC (0.976 +/- 0.011, P0.001) with a sensitivity of 77.6% - 91.4% and a specificity of 92.2% - 100%. Six mass peaks were combined by logistic regression to achieve the AUC 0.897 +/- 0.054, 0.978 +/- 0.021 and 0.792 +/- 0.107 (P0.05) in the three groups (patients at stage I and II, stage II and III, stage III and IV). One mass peak (M/Z 4,016) was screened (P0.05) significantly between the preoperative and postoperative PaCa samples and the intensity decreased weeks after operation.Decision tree and logistic regression classification models of the mass peaks screened by SELDI-TOF-MS serum profiling can be used to differentiate pancreatic adenocarcinoma from normal pancreas, and is superior to CA 199. The detected mass peaks are helpful for the evaluation of curative effect and prognosis of pancreatic adenocarcinoma.

The epithelium is an active participant in the host response to infection. We hypothesized that epididymal epithelia play a role in the innate immune responses by sensing the presence of pathogens, expressing and secreting inflammatory cytokines that recruit inflammatory cells in response to invading pathogens. Our results indicated that TNF-alpha and IL-1beta could be secreted by the primary cultured rat epididymal cauda epithelia infected with Staphylococcus aureus. Epididymal epithelial-induced nitric oxide synthase (iNOS) expression was up-regulated after S. aureus infection and nitric oxide (NO) was also found to be produced significantly. NF-kappaB inhibitor BAY11-7082 inhibited TNF-alpha secretion completely and p38 mitogen-activated protein kinases (MAPKs) inhibitor SB203580 decreased TNF-alpha secretion partly, indicating that NF-kappaB and p38 signal pathways were involved in this inflammation response. Toll-like receptor (TLR)-2 and -4 were shown to be expressed in primary cultured rat epididymal epithelia. After infection the level of TLR2 expression was up-regulated rather than TLR4. These results demonstrated that epididymal epithelium have an innate immune response through activation of p38 MAPK and NF-kappaB after TLR2 activation by S. aureus infection.

We describe an ultraviolet (UV) absorbance titration method that can be used to determine complexing capacities (C(L)) and conditional stability constants (logK) of humic substances (HSs) with metal ions such as Cu(II) and Hg(II). Two fulvic acids (FA) and one humic acid (HA) were used for this study. UV absorbance of HSs gradually increased with the addition of Cu(II) or Hg(II) after blank correction, and these increases followed the theoretical 1:1 (ligand:metal ion) binding model. The results from the absorbance titration calculation for HSs with Cu(II) and Hg(II) compared well with those from fluorescence quenching titration. The titration of the model compound l-tyrosine with Cu(II) proved the validity of this method, and the K and C(L) were within 2.3% and 7.4% of the fluorescence quenching titration. The results suggest that the UV absorbance titration can be used to study the binding capacities of HSs and/or dissolved organic matter (DOM) with trace metals. The advantages and disadvantages of the absorbance titration method were also discussed.

Sm–Nd isotopic system is a powerful tracer and dating tool in geochemistry and cosmochemistry. Thermal ionization mass spectrometry (TIMS) is the benchmark method for obtaining high precision Nd isotope ratios. Traditionally, a two-step separation was employed in order to obtain high purity Nd fraction, which makes the sample preparation time-consuming. In this study, we present a new method to directly obtain precise and accurate Nd isotope ratios of geological samples without Sm–Nd separation. The key point is to well correct isobaric interferences of 144Sm on 144Nd for this method. The accurate intensity of 144Sm can be obtained using 147Sm/144Sm ratio of 4.87090 and mass discrimination factor of Sm (βSm), which is achieved using 147Sm/149Sm ratio of 1.08583 with Russell equation. Consequently, the real intensity of 144Nd was achieved, then also raw 146Nd/144Nd, 143Nd/144Nd ratios. Finally, 143Nd/144Nd ratio is normalized using 146Nd/144Nd ratio of 0.7219 following exponential law. The accuracy of this method is validated by replicating TIMS measurements for thirteen international reference materials of silicate rock, spanning a wide range of Sm/Nd from 0.13 to 0.46 and bulk compositions. The measured 143Nd/144Nd ratios of the rock standards are in good agreement with the reported values of within error of ±0.004%. Our new method shortens the analytical procedure and significantly improves sample throughput which greatly contributes to those studies requiring a large dataset and quick analysis.