3-Methyl-4-nitrophenol Exposure Deteriorates Oocyte Maturation by Inducing Spindle Instability and Mitochondrial Dysfunction.

3-methyl-4-nitrophenol (PNMC), a well-known constituent of diesel exhaust particles and degradation products of insecticide fenitrothion, is a widely distributed environmental contaminant. PNMC is toxic to the female reproductive system; however, how it affects meiosis progression in oocytes is unknown. In this study, in vitro maturation of mouse oocytes was applied to investigate the deleterious effects of PNMC. We found that exposure to PNMC significantly compromised oocyte maturation. PNMC disturbed the spindle stability; specifically, it decreased the spindle density and increased the spindle length. The weakened spindle pole location of microtubule-severing enzyme Fignl1 may result in a defective spindle apparatus in PNMC-exposed oocytes. PNMC exposure induced significant mitochondrial dysfunction, including mitochondria distribution, ATP production, mitochondrial membrane potential, and ROS accumulation. The mRNA levels of the mitochondria-related genes were also significantly impaired. Finally, the above-mentioned alterations triggered early apoptosis in the oocytes. In conclusion, PNMC exposure affected oocyte maturation and quality through the regulation of spindle stability and mitochondrial function.

Removal of heavy metals in water-extracted solution through adsorption by palygorskite and stabilization by comilling.

Removing water-soluble chlorides (WSCs) through water extraction is a common pretreatment technology for recycling municipal solid waste incineration (MSWI) fly ash (FA). However, the extracted solution often contains heavy metals, the concentrations of which exceed standards for effluent. This study aims to investigate the adsorption of heavy metals by palygorskite in water-extracted solution and explore the feasibility of stabilizing heavy metals through comilling palygorskite-adsorbed heavy metals (PAHMs) with water-extracted fly ash (WFA). The experimental parameters include: two-stage water extraction with a liquid-to-solid ratio of 5, adding 0, 0.125, 0.25, 0.5, 1, 2 or 3 g of palygorskite to 100 mL of water-extracted solution, and comilling the mixture of PAHMs and WFA for 0, 0.5, 1, 2, 4, 8, 12, 24 or 96 hours. The experimental results revealed that 3 g of palygorskite in 100 mL of extracted solution could absorb Pb, Cd, Cr, Cu and Zn, meeting the effluent standards. The total amount of Pb, Cd, Cr, Cu and Zn removal rate reached 99.7%. Moreover, 98.44% of the WSCs were not adsorbed, the water extraction process for removing WSCs was not compromised. After the comilling of PAHMs and WFA, the distribution of the heavy metals in the milled blended powder was greater than 99.44%; moreover, toxicity characteristic leaching procedure concentrations were determined to conform to regulatory standards, and the sequential extraction procedure revealed that the heavy metals tended to be in stable fractions. This achieves the goal of preventing secondary pollution from heavy metals during the MSWI FA recycling process.

TET Family Members Are Integral to Porcine Oocyte Maturation and Parthenogenetic Pre-Implantation Embryogenesis.

The ten-eleven translocation (TET) enzyme family, which includes TET1/2/3, participates in active DNA demethylation in the eukaryotic genome; moreover, TET1/2/3 are functionally redundant in mice embryos. However, the combined effect of TET1/2/3 triple-gene knockdown or knockout on the porcine oocytes or embryos is still unclear. In this study, using Bobcat339, a specific small-molecule inhibitor of the TET family, we explored the effects of TET enzymes on oocyte maturation and early embryogenesis in pigs. Our results revealed that Bobcat339 treatment blocked porcine oocyte maturation and triggered early apoptosis. Furthermore, in the Bobcat339-treated oocytes, spindle architecture and chromosome alignment were disrupted, probably due to the huge loss of 5-hydroxymethylcytosine (5hmC)and concurrent increase in 5-methylcytosine (5mC). After Bobcat339 treatment, early parthenogenetic embryos exhibited abnormal 5mC and 5hmC levels, which resulted in compromised cleavage and blastocyst rate. The mRNA levels of EIF1A and DPPA2 (ZGA marker genes) were significantly decreased, which may explain why the embryos were arrested at the 4-cell stage after Bobcat339 treatment. In addition, the mRNA levels of pluripotency-related genes OCT4 and NANOG were declined after Bobcat339 treatment. RNA sequencing analysis revealed differentially expressed genes in Bobcat339-treated embryos at the 4-cell stage, which were significantly enriched in cell proliferation, cell component related to mitochondrion, and cell adhesion molecule binding. Our results indicated that TET proteins are essential for porcine oocyte maturation and early embryogenesis, and they act by mediating 5mC/5hmC levels and gene transcription.

Identification of miR-192 target genes in porcine endometrial epithelial cells based on miRNA pull-down.

INTRODUCTION: MicroRNAs (miRNAs)-a class of small endogenous non-coding RNAs-are widely involved in post-transcriptional gene regulation of numerous physiological processes. High-throughput sequencing revealed that the miR-192 expression level appeared to be significantly higher in the blood exosomes of sows at early gestation than that in non-pregnant sows. Furthermore, miR-192 was hypothesized to have a regulatory role in embryo implantation; however, the target genes involved in exerting the regulatory function of miR-192 required further elucidation. METHODS: In the present study, potential target genes of miR-192 in porcine endometrial epithelial cells (PEECs) were identified through biotin-labeled miRNA pull-down; functional and pathway enrichment analysis was performed via gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment. Bioinformatic analyses were concurrently used to predict the potential target genes associated with sow embryo implantation. In addition, double luciferase reporter vectors, reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and Western blot were performed to verify the targeting and regulatory roles of the abovementioned target genes. RESULTS: A total of 1688 differentially expressed mRNAs were identified via miRNA pull-down. Through RT-qPCR, the accuracy of the sequencing data was verified. In the bioinformatics analysis, potential target genes of miR-192 appeared to form a dense inter-regulatory network and regulated multiple signaling pathways, such as metabolic pathways and the PI3K-Akt, MAPKs, and mTOR signaling pathways, that are relevant to the mammalian embryo implantation process. In addition, CSK (C-terminal Src kinase) and YY1 (Yin-Yang-1) were predicted to be potential candidates, and we validated that miR-192 directly targets and suppresses the expression of the CSK and YY1 genes. CONCLUSION: We screened 1688 potential target genes of miR-192 were screened, and CSK and YY1 were identified as miR-192 target genes. The outcomes of the present study provide novel insights into the regulatory mechanism of porcine embryo implantation and the identification of miRNA target genes.

Pharmacological Agents That Prevent Postoperative Cognitive Dysfunction in Patients With General Anesthesia: A Network Meta-analysis.

BACKGROUND: Postoperative cognitive dysfunction (POCD) is associated with prolonged hospital stays, increased mortality, and negative socioeconomic consequences. Dexmedetomidine, ketamine, dexamethasone, and lidocaine have previously been reported to be effective for preventing POCD. STUDY QUESTION: In this network meta-analysis, we apply direct and indirect comparisons to rank these pharmacological agents in terms of their effect on POCD, through which we seek to provide evidence for future clinical medication. DATA SOURCES: A comprehensive literature search of PubMed, EMBASE, the Cochrane Library, and Web of Science was conducted to identify randomized controlled trials that examined the effects of dexmedetomidine, ketamine, dexamethasone, or lidocaine on POCD induced by general anesthesia. STUDY DESIGN: For eligible studies, 2 reviewers independently extracted data and assessed the respective risk of bias. Bayesian network meta-analysis was conducted using R-3.4.1 software. A total of 30 articles were included in this meta-analysis. RESULTS: Direct comparison showed that dexmedetomidine [odds ratio (OR) = 0.42, 95% confidence interval (CI): 0.32-0.55] may decrease the incidence of POCD for noncardiac surgery; dexmedetomidine (OR = 0.08, 95% CI: 0.01-0.63) and ketamine (OR = 0.09, 95% CI: 0.02-0.32) were found to decrease POCD for cardiac surgery. The R-software ranking result for prevention of POCD was dexmedetomidine, lidocaine, ketamine, placebo, and dexamethasone, respectively, in noncardiac surgery, and dexmedetomidine, ketamine, lidocaine, placebo, and dexamethasone in cardiac surgery. CONCLUSION: Dexmedetomidine exhibited obvious superiority to other agents for noncardiac surgery; dexmedetomidine and ketamine exhibited a significantly better performance than other agents for cardiac surgery. Dexamethasone did not show better efficacy than the placebo. However, more rigorously designed studies comprising larger sample sizes are needed to confirm our findings.

Propentofylline reduces mechanical allodynia and induces mitogen-activated protein kinase phosphatase-1: An experimental study in a rat model of acute incisional pain.

Background: Acute postoperative pain can lead to long hospital stays, dysfunction, and even chronic pain. Mitogen-activated protein kinases (MAPKs) are important in pain signaling. The activity of MAPKs are negatively regulated by dual specificity phosphatases such as mitogen- activated protein kinase phosphatase-1 (MKP-1), which specifically dephosphorylates MAPK p38. Since, propentofylline (PPF) can reduce pain by inhibiting MAPKs, we hypothesized that PPF relieves acute pain by inducing MKP-1 levels. Objective: Investigating the anti-nociception effect of an intrathecal injection of PPF in a rat model of acute incisional pain and the role of MKP-1 and phospho-p38 in the mechanism by which PPF ameliorates acute pain. Methods: We assessed the mechanical withdrawal threshold (MWT) response before and after incisional pain surgery between a control group and a group receiving PPF, and also assessed the effect of pre-treatment with Ro 31-8220, an MKP-1 inhibitor, on PPF effects. Following the MWT, lumbar spinal cord samples were also analyzed by western blot analysis to determine MKP-1 and p-p38 levels. Results: Following surgery, the MWT response was decreased over 5 h-3 d accompanied by decreased expression of MKP-1 and increased p-p38 levels. An intrathecal injection of PPF increased the MWT response and increased spinal cord MKP-1 expression, but decreased p-p38 levels. Pre-treatment of rats with Ro31-8220 partly reversed the analgesic effect of PPF and its effect on MKP-1/p-p38 levels. Conclusions: This study suggests that an increase in MKP-1 levels and a corresponding decrease in p-p38 levels may be the mechanism by which PPF ameliorates acute pain.

Clinicopathological and Prognostic Value of Ki-67 Expression in Bladder Cancer: A Systematic Review and Meta-Analysis.

BACKGROUND: Ki-67 is an established marker of cell proliferation, and the Ki-67 index correlates with the clinical course of several cancer types, including bladder cancer (BC). However, the clinicopathological and prognostic significance of Ki-67 in bladder cancer remains unclear. Therefore, we performed a systematic review and meta-analysis to clarify this relationship. METHODS: A comprehensive literature search for relevant studies published up to February 1, 2016, was performed using PubMed, Cochrane Library, Embase and ISI Web of Knowledge. The effects of Ki-67 expression on survival outcome in patients with BC and BC subtypes were evaluated. Furthermore, the relationship between Ki-67 expression and the clinicopathological features of BC were assessed. RESULTS: Thirty-one studies with 5147 bladder cancer patients were selected for evaluation. Ki-67 expression was significantly associated with shorter recurrence-free (HR 1.69, 95% CI: 1.33-2.14), progression-free (HR 1.89, 95% CI: 1.43-2.51), overall (HR 2.03, 95% CI: 1.31-3.16), and cancer-specific (HR 1.69, 95% CI: 1.47-1.95) survival. Moreover, whereas high expression was more common in high tumor stage, recurrence status, tumor size, there was no correlation between high Ki-67 expression and age, gender, smoking habits, and tumor number. Importantly, analysis of the different subgroups of BC suggested that significant correlations between high Ki-67 expression and survival outcome (recurrence-free/progression-free/overall/cancer-specific survival) are present only in European-American patients. CONCLUSION: The present results indicate that over-expression of Ki-67 is distinctly correlated with poor patient survival. Ki-67 may serve as a valuable biomarker for prognosis in BC patients, particularly in non-Asian BC patients. The results suggest no significant association between Ki-67 expression and BC prognosis in Asian patients. Further efforts are needed to fully clarify this relationship.

Effects of Chlorella sp. on nutrient treatment in cultures with different carbon to nitrogen ratios.

Chlorella sp. is often used in the treatment of wastewater to produce lipids, a practice which could go beyond wastewater treatment and be used to generate green energy. Our objectives here are to explore how the ratio of carbon to nitrogen (C/N) affects the removal of carbon and nitrogen in a wastewater treatment system, while simultaneously generating biomass and lipids. In this study, the C/N ratio is adjusted to 0.002, 6, 8, 10, 12 and 32. The results indicate that a C/N of 10 is sufficient to ensure the consumption of carbon and nitrogen, achieving the lowest concentration in the shortest culturing time (32 h). When nitrogen is lacking in the culture, there will be a slight decrease in the rate of carbon consumption which leads to a limitation of nitrogen and an increase in the lipid/cell density even at 96 h of culture time. The highest lipid content (0.57 g/L) and lipid increase rate (0.4 g/L) occurs with a C/N of 32. The greatest amount of biomass, 1.42 g/L is achieved when the C/N is 32. The carbon concentration is the main factor affecting the nitrogen consumption and the increase in the biomass and lipid content.

Improving the mechanical characteristics and restraining heavy metal evaporation from sintered municipal solid waste incinerator fly ash by wet milling.

The milling process has a verified stabilizing effect on the leaching of heavy metals into the environment from municipal solid waste incinerator (MSWI) fly ash. The aim of this current study is to further improve and confirm the effectiveness of the process by exploring its effects on the evaporation of heavy metals and on the mechanical characteristics of the sintered MSWI fly ash. The chemical composition of the MSWI fly ash is first altered by the addition of water treatment plant sludge (WTS) and cullet, and then processed to produce sintered specimens suitable for reuse as an aggregate. In the experiments, fly ash, WTS and cullet (40%: 30%: 30%, respectively) were mixed and milled for 1h. Samples were sintered for 60 min at temperatures of 850, 900, 950 and 1000°C. Test results confirm that milling increased the compressive strength of the sintered specimens. The compressive strength of unmilled specimens sintered at 900°C was only 90 kg/cm(2), but that of milled specimens was 298 kg/cm(2) when sintered at only 850°C. There was also an improvement in the soundness ranging from 11.04% to 0.02% and a reduction in the evaporation rates of Pb, Cd, Cu, Cr and Zn from 54-64%, 43-49%, 38-43%, 30-40% and 14-35% (900-1000°C) to 19-21%, 19-21%, 14-19%, 12-19% and 14-17% (850-1000°C), respectively. The improvement in compressive strength was attained by the combination in the liquid sintering stage of powdered ash with the amorphous material. The amorphousness of the material also helped to seal the surface of the fly ash, thereby reducing the evaporation of heavy metals during the heating process.

Effect of the milling solution on lead stabilization in municipal solid waste incinerator fly ash during the milling processes.

The wet milling process had been found to effectively stabilize lead in fly ash. This study adopts this method and looks at the effect of different milling solutions to improve the efficiency of lead stabilization. Different milling solutions (water, phosphoric acid and ethanol) and different milling times (1, 24, 48 and 96 h) were selected as parameters. Since lead oxide can be identified by XRD in this experiment's samples, 5% lead oxide was added to the extracted mixed fly ash to make instrumental analysis easier. The experimental results indicate that the effect of stabilization of lead after milling could exceed 96%. During milling with water, considerable lead leached into the water in the initial stage (1 h) of the process, but a stable level was reached as the milling time increased. After milling with ethanol and 0.2 M phosphoric acids, the efficiency could exceed 93% after 1 h of milling time. The results of the sequential extraction procedure (SEP) results show that the residual fraction could be increased from 8.93% to 56.16% when a 0.2 M phosphoric acid solution was used. Clearly the choice of an appropriate milling solution can enhance lead stabilization in the fly ash.

Roentgen stereophotogrammetric analysis and clinical assessment of unipolar versus bipolar hemiarthroplasty for subcapital femur fracture: a randomized prospective study.

BACKGROUND: Hemiarthroplasty is a well-established treatment for displaced subcapital fracture, but controversy exists about the optimal implant type. Bipolar hemiarthroplasty has proposed advantages over unipolar hemiarthroplasty in terms of better clinical results and decreased wear of acetabular cartilage. METHODS: This study is a randomized prospective study of 51 patients (52 hips) receiving either bipolar or unipolar hemiarthroplasty for displaced subcapital fractures. The outcome measurements were clinical scores and Roentgen stereophotogrammetric analysis (RSA) analysis to determine the rate of acetabular wear. RESULTS: Twenty-three patients completed 2-year follow-up. The RSA data demonstrated that there was slightly less acetabular wear by bipolar prostheses than by unipolar. The combined mean three-dimensional wear of the bipolar prostheses was 0.6 mm compared with 1.5 mm for the unipolar prostheses (P= 0.04). The bipolar group generally achieved higher scores in terms of the Harris Hip Score, Western Ontario and McMaster University Index of Osteoarthritis (WOMAC) questionnaire and 6-min walk test. These results were statistically significant at 3 months but not at 12 and 24 months. CONCLUSION: This study suggests that while the bipolar prosthesis performs slightly better than the unipolar in terms of acetabular cartilage wear and clinical outcomes, it remains debatable whether the benefits are worth the increased cost of the prosthesis.

Effects of wet ball milling on lead stabilization and particle size variation in municipal solid waste incinerator fly ash.

Water-extracted municipal solid waste incinerator (MSWI) fly ash was treated by a process of wet ball milling, using desalinated water as the milling solution. We investigated the influence of the milling process on the partitioning and leaching characteristics of lead (Pb) and the particle size distribution. The results show that 93.11% of the Pb was partitioned into the milled ash, 2.60% to the milling balls, and 0.17% to the inner surface of the milling jar, while amounts lower than the detection limit remained in the milled solution. As tested by the toxicity characteristic leaching procedure (TCLP), the leaching of Pb was inhibited after short-term grinding (from 5.2 to 1.2mg/L after 1h of milling), and further reduced by about 96% after 96h of ball milling. The mobility of the heavy metal was analyzed after a sequential extraction procedure. The results also show that Pb tended to become more stable after milling. The size distribution of particles was analyzed by a laser particle diameter analyzer and their morphology during grinding was observed using scanning electron microscopy. The median size of the fly ash decreased significantly from 36 to 5 microm after 0.5h of milling, but then only slightly, from 5 to 2 microm, with further milling from 0.5 to 96 h, due to the concurrent actions of fragmentation and/or agglomeration. The reason for the stabilization of Pb by ball milling was probably that Pb was sealed in the milled fly ash during the fragmentation and agglomeration of particles.

Capsular-enhanced repair of the short external rotators after total hip arthroplasty.

Higher dislocation rates have been reported with the posterior approach to the hip. Empirical studies suggest that careful repair of the posterior structures significantly reduces this risk. However, studies examining the integrity of repair using plain radiographs and metallic markers have reported high failure rates. To explain this discrepancy, we performed a study using radiostereometric analysis to assess the repair. Ten patients were recruited. Markers were placed into the capsule and bone. The capsule and conjoined short external rotators were repaired through drill holes in bone. At 3 months, stress radiostereometric analysis radiographs were taken in internal and external rotation. Eight of 10 patients had a mean of 3.51-mm difference in separation, suggesting that the repair was intact. We recommend careful repair of posterior structures when using the posterior approach to reduce the risk of dislocation.

Validation of a low-dose hybrid RSA and fluoroscopy technique: Determination of accuracy, bias and precision.

Analyzing skeletal kinematics with radiostereometric analysis (RSA) following corrective orthopedic surgery allows the quantitative comparison of different implant designs. The purpose of this study was to validate a technique for dynamically estimating the relative position and orientation of skeletal segments using RSA and single plane X-ray fluoroscopy. Two micrometer-based in vitro phantom models of the skeletal segments in the hip and knee joints were used. The spatial positions of tantalum markers that were implanted into each skeletal segment were reconstructed using RSA. The position and orientation of each segment were determined in fluoroscopy images by minimizing the difference between the markers measured and projected in the image plane. Accuracy was determined in terms of bias and precision by analyzing the deviation between the applied displacement protocol and measured pose estimates. Measured translational accuracy was less than 100 microm parallel to the image plane and less than 700 microm in the direction orthogonal to the image plane. The measured rotational error was less than 1 degrees . Measured translational and rotational bias was not statistically significant at the 95% level of confidence. The technique allows real-time kinematic skeletal measurements to be performed on human subjects implanted with tantalum markers for quantitatively measuring the motion of normal joints and different implant designs.