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Summary As global warming intensifies, emerging evidence has demonstrated high ambient temperature during pregnancy negatively affects maternal physiology with compromised pregnant outcomes; however, little is known about the roles of gut microbiota and its underlying mechanisms in this process. Here, for the first time, we explored the potential mechanisms of gut microbiota involved in the disrupted glycolipid metabolism via hepatic mitochondrial function. Our results indicate heat stress (HS) reduces fat and protein contents and serum levels of insulin and triglyceride (TG), while increases that of non‐esterified fatty acid (NEFA), β‐hydroxybutyric acid (B‐HBA), creatinine and blood urea nitrogen (BUN) (P

Jianwen He,* Yunhua Mao,* Wentao Huang,* Mingzhao Li, Huimin Zhang, Yunhao Qing, Shuo Lu, Hengjun Xiao, Ke Li Department of Urology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ke Li Email like35@mail.sysu.edu.cnHengjun Xiao Email hjxiao555@126.comPurpose: Prostate cancer (PCa) is the most common cancer in American men, and the mechanisms of development and progression are still not completely clear. Methylcrotonoyl-CoA carboxylase 2 (MCCC2) was previously identified overexpressed in PCa with lymph node metastasis, but its specific role and mechanisms need further investigation. This study aimed to investigate the role of MCCC2 in PCa cells and its underlying mechanisms.Materials and Methods: Quantitative RT-PCR and Western blotting were used to detect MCCC2 mRNA and protein expression in normal prostate epithelium and cancerous cells. Upon manipulation of MCCC2 expression, cell proliferation was measured by CCK-8 assays and migration and invasion were determined by transwell assays. Changes of apoptosis, cell cycle and mitochondrial membrane potential were evaluated by flow cytometry. MCCC2-mediated signaling pathways were screened by bioinformatics and verified by RT-PCR and Western blotting. Finally, immunohistochemistry was performed to detect the expression of MCCC2 and glutamate dehydrogenase 1 (GLUD1) in PCa tissues to analyze their correlation.Results: We demonstrated that MCCC2 promoted cell proliferation, migration and invasion but inhibited apoptosis in PCa cells. In addition, MCCC2 in 22Rv1 cells induced mitochondrial damage. In PCa tissues, MCCC2 overexpression associated with lymph node metastasis (P=0.001) and high Gleason scores (P< 0.001). MCCC2 positively correlated with GLUD1 expression in PCa tissues (r=0.435, P< 0.001). Ectopic overexpression of MCCC2 up-regulated GLUD1 and p38 MAPK expression, whereas inhibition of MCCC2 decreased GLUD1 and p38 MAPK expression.Conclusion: MCCC2 exerts oncogenic function in PCa through regulating GLUD1-p38 MAPK signaling pathway, and it may be a potential treatment target.Keywords: prostate cancer, MCCC2, GLUD1, p38 MAPK

The placenta is critical for the regulation of fetal innate immune function. Maternal heat stress (HS) impairs the immune function and the intestinal barrier in the offspring. However, the effects of maternal HS on the placental immune response and the development of the fetal intestine and its innate immune system remain unclear. Fetal mice were divided into the utero control (IUTN) and heat stress (IUHS) groups according to the maternal ambient temperature. Transcriptome analysis revealed that the expressions of placental immune response–related genes such as macrophage antigen CD68 and Fc gamma receptors 1 and 3 (fcgγ1 and fcgγ3) were increased, but the mRNA expression and protein levels of colony-stimulating factor-1 (Csf1) were decreased in the HS group compared with the TN group (p < 0.05). Furthermore, there was no significant difference in the intestinal length normalized to pup weight between the IUTN and IUHS groups. The expression of genes (such as alpi and ttr) involved in fetal duodenum and jejunum development was downregulated by maternal HS, whereas the expression of genes enriched in the cell cycle was increased. The mRNA expression and protein levels of cell division cycle 6 (Cdc6) in the fetal duodenum and jejunum were much higher in the IUHS group than in the IUTN group (p < 0.05). Maternal HS also down-regulated the expression of genes enriched in the innate immune system in the fetal duodenum and jejunum. The mRNA expression and protein levels of interleukin 1 alpha (IL1a) were reduced in the IUHS group compared with the IUTN group (p < 0.05). Taken together, these data demonstrated that maternal HS modulated the expression of genes in the placenta related to the immune response and inhibited the development of the fetal intestine and its innate immune system.

Nowadays, the increasing costs of law enforcement have become a considerable problem for the traffic department. With the insufficient police force, the department cannot separate the daily statistics of traffic flow at the intersection. In this case, understanding the daily traffic flow at the intersection helps to make the traffic light timing scheme, plan the path direction, understand the urban congestion, and so on. Therefore, we propose a vehicle detection model which can be deployed in NVIDIA Jetson AGX Xavier and is based on TensorFlow. To improve the accuracy and the speed of detection, we use CenterNet, a keypoint-base network model, and the backbone is the ResNet50. Hence, we can extract features of the image and use regression to predict the box of any object. The experimental results show that: the accuracy of vehicle flow identification is up to 98.23%, small vehicles 98.30%, medium vehicles 98.64%, and small vehicles 97.75% in a straight-ahead environment, meet the actual use requirements.

With the rapid growth of urbanization in China, traffic congestion, excessive exhaust emissions and energy consumption are becoming increasingly serious. This paper proposes a single intersection traffic signal timing optimization method based on sparrow algorithm. The four objectives of the road level traffic index data (Travel Time Index, TTI), traffic delay time, queue length and vehicle average carbon emissions are optimized. The sparrow algorithm is used to solve the timing of single intersection signal lights, and the VISSIM simulation platform is secondary developed to verify the energy-saving benefits of the method. The simulation platform test shows that, under the single intersection traffic signal timing optimization scheme, the average queue length of the intersection decreases by 26.26% compared with the non-active state, the delay time decreases by 27.64%, the intersection throughput increases by 13.94%, the TTI index increases by 43.4%, and the average carbon emission decreases by 31.89%, which has a significant effect on reducing the emission of single intersection.

High ambient temperature-induced heat stress (HS) during pregnancy may affect the placental function and fetal development. Late gestation is a critical period of the developing fetal brain and intestine. The study aimed to investigate the effects of HS during late pregnancy on the function of placenta, fetal brain and intestine in a mouse model. We found that the number of stillborn fetal mice were increased due to maternal HS. Transcriptome analysis revealed that the expression of genes enriched in nutrients transport and metabolism of HS group were up-regulated in the placenta, but down-regulated in the fetal duodenum and jejunum. Interestingly, the concentration of triglyceride (TG) in the HS group was raised in the placenta, but reduced both in the fetal duodenum and jejunum compared with the thermal-neutral (TN) group. Additionally, maternal HS also reduced total cholesterol (TC) contents in the fetal duodenum. The mRNA expression and protein levels of placental fatty acid binding protein 2 and 4 (fabp2 and fabp4) were not affected by maternal HS, but the mRNA expression and protein levels of cluster of differentiation 36 (CD36) and diacylglycerol acyltransferase-2 (Dgat2) were decreased in the fetal intestine. Furthermore, maternal HS reduced the mRNA expression and protein levels of the placental 11beta-hydroxysteroid dehydrogenase type 2 (Hsd11b2) and 5-hydroxytryptamine receptor 1D (Htr1d). The concentrations of corticosterone and the expression of heat shock protein 90 beta family member 1 (hsp90b1), hypoxia up-regulated 1 (hyou1) and corticotropin releasing hormone receptor 1 (crhr1) enriched in response to glucocorticoids in the fetal brain were increased by maternal HS. Taken together, our findings demonstrated that maternal HS disrupted the placental glucocorticoid barrier and serotonin system associated with the raised corticosterone levels in the fetal brain, which might contribute to the decreased capacity of nutrients transport and metabolism in the fetal intestine.

A reversible lane control system with low emission load is designed based on VISSIM simulation software. This system aims at the problem that the traffic flow of different directions at the intersection has the imbalance of time and space. Through the VISSIM simulation based on Python language, the second lane from the central separation belt is taken as the variable lane, and the lane control under different traffic flow conditions is realized. The system takes the queuing length, traffic delay time and carbon emissions of the intersections as the traffic efficiency evaluation indexes of the variable lane, and analyse the actual benefits of setting variable lanes to relieve the time and space resources of the intersections. Simulation results by using VISSIM and the designed Python interface show that, compared with the settings without variable lane control system, the average queue length at the intersection decreases by 38.6%, the maximum queue length by 36.4%, the average delay time by 20.7%, the maximum delay time by 15.1%, and the average carbon emission by 17.3%. It can be seen that the variable lane control system at intersections based on VISSIM has a significant effect on easing traffic congestion at intersections and reducing total carbon emissions.

In early 2020, novel coronavirus pneumonia broke out. In order to prevent the spread of the disease, governments around the world asked the masses to wear masks. However, there are still many people who do not wear masks in public places. To solve this problem, this paper proposes a mask detection device based on yolo3 framework. The device uses the yolov3 algorithm to extract the face prediction area, and uses the gray image to calculate the skin exposure rate of the mouth and nose of the face, so as to judge whether the recognized person is wearing a mask or not and whether the mask is wearing correctly. The model is deployed on the hardware to facilitate the staff to carry the detection. The experimental results show that the recognition rate is 86.6%.

Stress shows both direct- and indirect-effects on the functions of the gastrointestinal tract, in particular on the mucus physiology and the composition of microbiota. Mucus mainly consists of heavily glycosylated proteins called mucins, which are secreted by goblet cells. The gut mucus layer is a pivotal part of the intestinal protection and colonized by commensal microbes, essential for the development and health of the host. There is a symbiotic interaction between intestinal microbiota and the host cells. On the one hand, mucus provides nutrients for the growth and adhesion of microbes; on the other hand, mucin-degrading bacteria generate energy sources for the host epithelium. However, the mucusmicrobial interaction has rarely been considered in the context of stress exposure. Therefore, this paper principally reviews the effects of stress on both mucus secretion and gut microbiota and is hoped to provide a new perspective for future study.

Metastasis is the main cause of cancer-specific death in patients with prostate cancer (PCa). Acyl-coenzyme A synthetase long-chain family member 3 (ACSL3) is involved in the metabolic reprogramming of multiple types of cancer cells, but its role in PCa metastasis remains largely unknown. Here, we determined the effect of overexpression or small interfering RNA-mediated depletion of ACSL3 on the migratory and invasive abilities of human PCa cell lines. We also conducted phospho-protein microarray analysis to identify signaling pathway components affected by ACSL3 modulation. Overexpression of ACSL3 promoted the migration and invasion of PCa cells, whereas ACSL3 downregulation had the opposite effects. Mechanistically, phospho-protein analysis showed that ACSL3 regulated the phosphorylation of AKT and the expression of matrix metalloproteinase9. Our results support a potential role for ACSL3 in promoting the metastatic behavior of PCa, possibly via AKT/matrix metalloproteinase9 pathways. Thus, ACSL3 could be a novel target for the development of treatments for PCa. Key words: ACSL3; AKT; matrix metalloproteinase9; metastasis; prostate cancer

Background Heat stress (HS) jeopardizes intestinal barrier functions and augments intestinal permeability in pigs. However, whether HS-induced maternal microbial and metabolic changes in primiparous sows during late gestation remains elusive. We present here, a study investigating the fecal microbial and metabolic responses in late gestational primiparous sows when exposed to HS. Methods Twelve first-parity Landrace × Large White F1 sows were randomly assigned into two environmental treatments including the thermoneutral (TN) (18–22 °C; n = 6) and HS (28–32 °C; n = 6) conditions. Both treatments were applied from 85 d of gestation to farrowing. The serum and feces samples were collected on d 107 of gestation, for analyses including intestinal integrity biomarkers, high-throughput sequencing metagenomics, short-chain fatty acid (SCFA) profiles and nontargeted metabolomics. Results Our results show that HS group has higher serum Heat shock protein 70 (HSP70), lipopolysaccharide (LPS) and lipopolysaccharide-binding protein (LBP) levels. The gut microbial community can be altered upon HS by using β-diversity and taxon-based analysis. In particular, the relative abundance of genera and operational taxonomic units (OTUs) related to Clostridiales and Halomonas are higher in HS group, the relative abundance of genera and OTUs related to Bacteroidales and Streptococcus, however, are lower in HS group. Results of metabolic analysis reveal that HS lowers the concentrations of propionate, butyrate, total SCFA, succinate, fumarate, malate, lactate, aspartate, ethanolamine, β-alanine and niacin, whereas that of fructose and azelaic acid are higher in HS group. These metabolites mainly affect propanoate metabolism, alanine, aspartate and glutamate metabolism, phenylalanine metabolism, β-alanine metabolism, pantothenate and CoA biosynthesis, tricarboxylic acid cycle (TCA) and nicotinate and nicotinamide metabolism. Additionally, correlation analysis between significant microbes and metabolites indicated that the HS-induced microbiota shift is likely the cause of changes of intestinal metabolism. Conclusions Taken together, we reveal characteristic structural and metabolic changes in maternal gut microbiota as a result of late gestational HS, which could potentially provide the basis for further study on offspring gut microbiota and immune programming.

Heat stress (HS) during gestation has been associated with negative outcomes, such as preterm birth or postnatal metabolic syndromes. The intestinal microbiota is a unique ecosystem playing an essential role in mediating the metabolism and health of mammals. Here we hypothesize late gestational HS alters maternal microbial transmission and structures offspring's intestinal microbiota and serum metabolic profiles. Our results show maternal HS alters bacterial β-diversity and composition in sows and their piglets. In the maternal intestine, genera Ruminococcaceae UCG-005, [Eubacterium] coprostanoligenes group and Halomonas are higher by HS (q 0.05), whereas the populations of Streptococcus, Bacteroidales RF16 group_norank and Roseburia are decreased (q 0.05). In the maternal vagina, HS mainly elevates the proportions of phylum Bacteroidetes and Fusobacteria (q 0.05), whereas reduces the population of Clostridiales Family XI (q 0.05). In the neonatal intestine, maternal HS promotes the population of Proteobacteria but reduces the relative abundance of Firmicutes (q 0.05). Moreover, the core Operational taxonomic units (OTU) analysis indicates the proportions of Clostridium sensu stricto 1, Romboutsia and Turicibacter are decreased by maternal HS in the intestinal and vaginal co-transmission, whereas that of phylum Proteobacteria and Epsilonbacteraeota, such as Escherichia-Shigella, Klebsiella, Acinetobacter, and Comamonas are increased in both the intestinal and vaginal co-transmission and the vagina. Additionally, Aeromonas is the only genus that is transmitted from environmental sources. Lastly, we evaluate the importance of neonatal differential OTU for the differential serum metabolites. The results indicate Acinetobacter significantly contributes to the differences in the adrenocorticotropic hormone (ACTH) and glucose levels due to HS (P 0.05). Further, Stenotrophomonas is the most important variable for Cholesterol, low-density lipoprotein (LDL), diamine oxidase (DAO), blood urea nitrogen (BUN) and 5-hydroxytryptamine (5-HT) (P 0.10). Overall, our data provides evidence for the maternal HS in establishing the neonatal microbiota via affecting maternal transmission, which in turn affects the maintenance of metabolic health.

Late gestation is a key period for intestinal development. Maternal heat exposure may induce intestinal dysfunction of offspring. To investigate the responses of intestinal morphology and function of offspring to the maternal heat stress (HS), twelve first-parity Landrace × Large White sows were assigned to thermoneutral (TN) (18-22 °C; n = 6) or HS (28-32 °C; n = 6) treatment groups at 85 d of gestation until natural farrowing. Twenty-four newborn piglets (two piglets at medium body weight from each litter) were randomly selected and divided into in utero thermoneutral (IUTN, n = 12) and heat-stressed (IUHS, n = 12) groups according to the sow’s treatment. Blood and intestinal samples were harvested to evaluate stress hormone levels, intestinal morphology, integrity and barrier function in the newborn piglets. Our results showed that maternal HS piglets exhibited increased serum adrenocorticotropic hormone (ACTH) concentration compared with that observed in the IUTN group. IUHS piglets showed lower lactase activities in the jejunum and ileum, whereas no significant differences were found between the two groups in the length of intestine, villus length or crypt depth. Serum diamine oxidase (DAO) activity was increased in IUHS piglets. IUHS piglets also exhibited decreased ZO-1, ZO-2 and MUC2 mRNA expression in the jejunum, while the protein levels were not affected. Additionally, IUHS piglets had a lower apoptotic percentage and FAS mRNA expression in the jejunum than those in the IUTN group. Taken together, these results demonstrate that high ambient temperature during late gestation of primiparous sows causes stress response in neonatal piglets, compromising intestinal permeability and mucosal barrier function, which may be partly mediated by inducing intestinal apoptosis.

INTRODUCTION AND OBJECTIVE:Circulating tumor cells (CTCs) is considered as “liquid biopsy” for tumor prognosis, and the detection of programmed death-ligand 1 (PD-L1) expressing in CTCs could promo...

Due to the supply-side reform and environmental protection in China, many small coal mines have been closed since 2015. However, acid mine drainage from these coal mines are continuously discharging into many rural creeks, which requires the systematical investigation on the variations of geochemical and environmental biological aspects in these water systems. In this study, from a classic acid mine drainage (AMD) from a closed coal mine of Hunan, China, various sediments and water samples in different sections were collected and analyzed. According to the corresponding Mineralogical and simple bacterial characteristics analysis (16S rRNA gene sequencing), the main findings were: 1) Secondary iron-containing minerals gradually transited from Gr(CO32−) (green rust), Sh (schwertmannite) to Akg (Akaganeite) and more stable Gt (Goethite); 2) compared to the pristine sediment, these minerals decreased the acid-neutralizing capacity and cation exchange capacity (CEC) of sediments; 3) Proteobacteria and Firmicutes were the dominant phyla and the obvious variation of Firmicutes species was observed in the creek affected by AMD, which probably could been a biological index to diagnose the natural attenuation of AMD. These results could be greatly significant to understand typical variations of creek attenuation and bacterial community in the presence of high metal and sulfate concentration.

Heat stress (HS) alters metabolic parameters and reduces productive performance in lactating sows. However, the impact of HS on metabolomic profiles of sows during late gestation is not fully understood. We present here, a study investigating the productive performance and metabolic responses in sows when exposed to HS during late gestation. Twelve first-parity Landrace × Large White F1 sows were randomly assigned into two environmental treatments including the thermoneutral (TN) (18-22 °C; n = 6) and HS (28-32 °C; n = 6) conditions from 85 d of gestation until farrowing. Rectal temperature (RT), respiration rates (RR), and surface temperature (ST) were measured every 4 h from 0800 h to 2000 h during the 2nd week. Farrowing and litter Data, as well as duration of eating, were monitored to assess sows' productive performance. Blood biochemical parameters and urinary metabolomic profiles were measured on d107 of gestation to analyze the host metabolic responses. Our results show that HS increased RT, RR, and ST (P 0.0001). Duration of parturition was prolonged during the delivery in HS group (P 0.05). Piglet body weight (BW) at d 10 and weaning were reduced by 18% and 17% respectively due to maternal HS (P 0.001). Duration of eating increased as a result of HS (P 0.001), consistent with the significant changes observed in serum ghrelin (P 0.05). Moreover, serum ACTH, cortisol, insulin, creatinine, and BUN saw increase as well (P 0.05). Plasma NEFA were elevated by HS (P 0.001). Additionally, HS elevated (VIP1, log

We evaluated the effect of cold shock on the fatty acid composition, antioxidant enzymes, and physiological responses of genetically improved farmed tilapia (GIFT). Experimental GIFT tilapia, a warm-water teleost, were initially acclimated at 28°C and then transferred directly to 13°C. Stress responses were monitored for 120h. There was a significant change in all parameters in response to the cold stressor (P

This chapter provides guidelines on immunoassay pre-selection screening and cost-effective initial assay evaluation.

This chapter describes the principle features of qualitative and semi-quantitative immunoassays. Performance characteristics and other regulatory requirements related to clinical application of qualitative immunoassays are explained. The assay design, optimization, calibration, validation and verification steps are described.

The fundamental requirements for an ELISA under development are discussed. ELISA development is presented as a series of steps, starting with understanding the analyte. Options for assay format are reviewed next. Antibody (or antigen in tests for antibody) selection is reviewed in detail, including monoclonal vs. polyclonal antibody properties and capture vs. detection antibody requirements. Microtiter plate coating and blocking, and separation and washing, are explained. In the signal generation and detection section, the simplest option of using a secondary antibody is described. The selection of enzyme and signal type is reviewed. Assay optimization is explained, including adjustment of incubation time, selection of appropriate buffers, blocking, formulation, titration, and sample volume to optimize the assay kinetics and minimize interferences. Hardware, software and curve-fitting are briefly mentioned. Assay management considerations (standardization, calibration, quality control and validation) are reviewed. Finally, ELISA tips and common troubleshooting situations are listed.

This is a very comprehensive troubleshooting guide for all types of immunoassays. Instructions are provided on use of the chapter to investigate and improve unsatisfactory assay performance, through root cause analysis. Each type of performance issue has a separate section, with a list of possible causes that can be used as a starting point. For each possible cause, information is provided to allow the reader to dismiss unlikely causes immediately, resulting in a shortlist of possible causes that are consistent with the facts. Then a suggested course of action provides a strategy for identifying the most likely cause(s) from the shortlist. Recognizing that the investigation of many immunoassay problems involves a user and a manufacturer working together, a checklist of useful information is provided for the user to prepare for an efficient dialogue with the manufacturer. Finally a case study for each type of problem is provided, with the eventual solution.

The study was aimed to examine the effects of exogenous 17β-estradiol (E2) on oocyte nest breakdown and follicular growth in the ovary of neonatal and immature mice. The effect of E2 on primordial follicle formation and differentiation were examined by treating neonatal mice with E2 in mineral oil at a dose of 10 mg/kg by sc injection each day from postnatal days 1 (PND1) to 10 (PND10). This investigation was conducted by using histological observation and immunohistochemical staining of proliferating cell nuclear antigen (PCNA); the numbers of follicles at each stage were examined to assess the follicular development. The results indicated that such dose of E2 inhibited oocyte nest breakdown from PND1 to PND10 significantly and enhanced the oocyte staining intensity of PCNA in the ovary by PND3, 5 and 10. PCNA staining was mainly detected on growing follicles (stages later than primary follicle). Our results also revealed that E2 decreased the size of the reproductive tract at PD10. This dose of E2 treatment inhibits oocyte nest breakdown and primordial follicle assembly, and causes follicle losses after PND5. Effects of exogenous E2 on follicular development in the neonatal and immature mouse may through a PCNA-dependent pathway.

As offshore production shifts to deep water, riser systems become more vulnerable. Riser clashing is a critical issue needed to be addressed in the design stage. Two design principles are defined in the code and termed as no collision allowed and collision allowed. Hydrodynamic interference is found to be the main driver of riser collision. This thesis proposes practical approaches to evaluate the probability of failure due to riser collision. With this important parameter, designers can optimally determine the riser layout. The whole work is separated into three stages. Doctor of Philosophy (CEE)