The purpose of the study was to study the activity of the phytoestrogen genistein (GEN) act- ing on FSHR and LHR in rat ovaries with polycystic ovary syndrome (PCOS). Sixty rats were di- vided into six groups. Rats in the dose group received genistein at a concentration of either 5 (low genistein dose group, L-gen), 10 (middle genistein dose group, M-Gen) or 20 (high genistein dose group, H-Gen) mg per kg of body weight per day. Estrogen group (EG, received 0.5 mg/kg Dieth- ylstilbestrol). Concentration of sex hormones in serum was quantified by enzyme-linked immuno- sorbent assay (ELISA). Expressions of follicle-stimulating hormone receptor (FSHR) and lutein- izing hormone receptor (LHR) protein were determined by immunohistochemistry. Treatment with genistein resulted in a strong stimulation of the concentration of sex hormone in serum. The concentration of progesterone and FSH was signiﬁcantly higher in H-Gen when compared to the PCOS model control group (MG) (P < 0.01). In contrast, the concentration of testosterone, LH and the ratio of LH/FSH decreased in GEN treatment groups compared to MG, the effect was statistically significant, tested by the ANOVA test (p<0.01). For hormone receptor activity, treat- ment with genistein resulted in an improvement of ovarian function with LHR protein expression being enhanced and FSHR protein expression being suppressed. Our results demonstrate that Genistein played a significant role in regulating FSH and LH receptor expression to improve perimenopausal ovarian and uterine function.
In this investigation, the effects of genistein (GEN) on the expression of steroidogenic genes such as steroidogenic acute regulatory protein (StAR), side-chain cleavage enzymes (P450scc) and cytochrome P450 aromatase (CYP19) were assessed. For this study, forty young female Sprague Dawley (SD) rats at aged 2-3 months (200±20 g) and forty aged female SD rats aged 10-12 months (490±20 g) were selected. Also, based on weight they were divided into a negative control group (NC), three different GEN dose groups, which received GEN of 15, 30, 60 mg/kg, and a positive control group (PC). The experiment lasted 30 days. Concentrations of serum hormones were determined by Enzyme-linked immunosorbent assay (ELISA). Gene and protein expressions of StAR, P450scc and CYP19 were determined by Real-Time PCR and western blot techniques. It was observed that 30-60 mg/kg GEN could increase the expression of androgen generating key enzymes in the young rat ovary. GEN also significantly increased progesterone and E2 levels in the serum of aged rats and reduced the levels of LH and FSH in the serum of both young and aged rats. Compared with young rats, the effect of GEN on the ovary of aged rats was stronger and a lower dose of GEN (15 mg/kg) showed an obvious effect on these indicators. GEN influenced both estrogen level and indicators associated with estrogen and androgen transformation processes, which indicates that GEN can impair the growth and maturation of the ovary.
Insulin receptor substrate 2 (IRS-2) modulates the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, which controls the suppression of gluconeogenic genes; IRS-2 is also a critical node of insulin signaling. Because of the high homology between pig and human IRS-2, we investigated the expression pattern and function of porcine IRS-2. QPCR and immunoblotting were used to detect the IRS-2 expression level in different tissues. There were high IRS-2 levels in the cerebral cortex, hypothalamus, and cerebellum in the central nervous system. In peripheral tissues, IRS-2 was expressed at relatively high levels in the liver. Immunohistochemistry analysis revealed that IRS-2 was mainly distributed in the hypothalamus and cerebral cortex. Furthermore, IRS-2 knockdown porcine hepatocytes and porcine aortic endothelial cells (PAECs) were generated. The IRS-2 knockdown induced abnormal expression of genes involved in glycolipid metabolism in hepatocytes and reduced the antiatherosclerosis ability in PAECs. In addition, we disrupted IRS-2 in porcine embryonic fibroblasts (PEFs) using the CRISPR/Cas9 genome editing system, before finally generating IRS-2 knockout embryos by somatic cell nuclear transfer (SCNT). Taken together, our results indicate that IRS-2 might be a valuable target to establish diabetes and vascular disease models in the pig.
The welfare and healthy growth of poultry under intensive feeding conditions are closely related to their living environment. In spring, the air quality considerably decreases due to reduced ventilation and aeration in cage systems, which influences the meat quality and health of broilers during normal growth stages. In this study, we analyzed the airborne bacterial communities in PM2.5 and PM10 in cage broiler houses at different broiler growth stages under intensive rearing conditions based on the high-throughput 16S rDNA sequencing technique. Our results revealed that PM2.5, PM10 and airborne microbes gradually increased during the broiler growth cycle in poultry houses. Some potential or opportunistic pathogens, including Acinetobacter, Pseudomonas, Enterococcus, Microbacterium, etc., were found in the broiler houses at different growth stages. Our study evaluated variations in the microbial communities in PM2.5 and PM10 and potential opportunistic pathogens during the growth cycle of broilers in poultry houses in the spring. Our findings may provide a basis for developing technologies for air quality control in caged poultry houses.
An integrated Z-source inverter for the single-phase single-stage grid-connected photovoltaic system is proposed in this paper. The inverter integrates three functional blocks including maximum-power-point-tracking, step-up/down DC-side voltage and output grid-connected current. According to the non-minimum-phase characteristic presented in DC-side and the functional demands of the system, two constant-frequency sliding-mode controllers with integral compensation are proposed to guarantee the system robustness. By using two controllers, the effects caused by the non-minimum-phase characteristic are mitigated. Under the circumstance of that the input voltage or the grid-connected current changes suddenly, the notches/protrusions following the over-shoot/ under-shoot of the DC-bus voltage are eliminated. The quality of grid-connected current is ensured. Also, a small-signal modelling method is employed to analyze the close-loop system. A 300W prototype is built in the laboratory. A solar-array simulator (SAS) is used to verify the systematic responses in the experiment. The correctness and validity of the inverter and proposed control algorithm are proved by simulation and experimental results.
Considering concrete nonlinearity, the wave height limit between small and large amplitude sloshing is defined based on the Bernoulli equation. Based on Navier-Stokes equations, the mathematical model of large amplitude sloshing is established for a Concrete Rectangle Liquid-Storage Structure (CRLSS). The results show that the seismic response of a CRLSS increases with the increase of seismic intensity. Under different seismic fortification intensities, the change in trend of wave height, wallboard displacement, and stress are the same, but the amplitudes are not. The areas of stress concentration appear mainly at the connections between the wallboards, and the connections between the wallboard and the bottom.
Compared with the robots, humans can learn to perform various contact tasks in unstructured environments by modulating arm impedance characteristics. In this article, we consider endowing this compliant ability to the industrial robots to effectively learn to perform repetitive force-sensitive tasks. Current learning impedance control methods usually suffer from inefficiency. This paper establishes an efficient variable impedance control method. To improve the learning efficiency, we employ the probabilistic Gaussian process model as the transition dynamics of the system for internal simulation, permitting long-term inference and planning in a Bayesian manner. Then, the optimal impedance regulation strategy is searched using a model-based reinforcement learning algorithm. The effectiveness and efficiency of the proposed method are verified through force control tasks using a 6-DoFs Reinovo industrial manipulator.
Objective: This study aimed to investigate developmental changes of the thymus and intra- thymic IL-1β, IL-6 and TNF-α expression in weaned Sprague-Dawley rats induced by lipopolysac- charide. Methods: Forty healthy weaned rats aged 26 days and weighing 83±4 g were randomly and equally divided into two groups. The lipopolysaccharide group was treated daily with a single injection of lipopolysaccharide for 10 consecutive days, and the saline group was treated with an equal volume of sterilized saline. On the 1st, 4th, 7th and 10th day, histological changes and distribu- tion of IL-1β-, IL-6- and TNF-α-positive cells were detected in the thymus by hematoxylin-eosin and immunohistochemistry staining, respectively. Subsequently, the expression levels of IL-1β, IL-6 and TNF-α were evaluated in the thymus by the ELISA method. Results: Thymus weight and index were significantly smaller in lipopolysaccharide-treated rats than in saline-treated rats (p<0.05), but no substantial changes were found in the thymus microstructure after lipopolysaccharide induction. Moreover, a large number of IL-1β-, IL-6- and TNF-α-positive cells were observed with brownish-yellow color and mainly distributed in the thy- mus parenchyma, both integrated optical density and average optical density increased signifi- cantly in lipopolysaccharide-treated rats than those in saline-treated rats. Compared with the saline group, most of the thymic homogenates had higher levels of IL-1β, IL-6 and TNF-α in the lipopolysaccharide group on different days. Conclusion: These findings indicate that the thymus atrophied after lipopolysaccharide induction in weaned Sprague-Dawley rats, and excessive production of intrathymic IL-1β, IL-6 and TNF-α was probably involved in the atrophic process.
Twinned dendrites in Al-Zn alloy with high Zn content (40% wt.%) were successfully prepared by directional solidification. At different directional solidification rates (1000 and 1500 μm/s), microstructures and growth orientation variations of Al twinned dendrite and non-twinned dendrite were characterized. By using the inverted trapezoidal graphite sleeve at 1000 μm/s, Al twinned dendrite were formed to developed feather crystal structures in longitudinal section. Its primary and secondary twinned dendrite were grew along  direction. Moreover the deviation angle between  direction of Al twinned dendrite and the heat flow direction was about 27.15°. While not using the inverted trapezoidal graphite sleeve at 1000 and 1500 μm/s, Al dendrite was the non-twinned dendrite and the twinned dendrite was not appeared. The experimental results showed that the higher temperature gradient, a certain pulling rate and convection environment were the formation conditions of twinned dendrites.
Culture gas atmosphere is one of the most important factors affecting embryo development in vitro. The main objective of this study was to compare the effects of CO concentration on the subsequent pre-implantation developmental capacity of pig embryos in vitro, including embryos obtained via parthenogenesis, in vitro fertilization (IVF), and intracytoplasmic sperm injection (ICSI). Pig embryos were developed in four different CO2 concentrations in air: 3%, 5%, 10%, or 15%. The cleavage rate of pig parthenogenetic, IVF, or ICSI embryos developed in CO2 concen- trations under 5% was the highest. There were no significant differences in the oocyte cleavage rate in ICSI embryos in CO2 concentrations under 3% and 5% (p>0.05). However, as CO2 levels increased (up to 15%) the blastocyst output on day 7, from parthenogenetic, IVF, and ICSI em- bryos, decreased to 0%. These findings demonstrate that CO2 positively affects the developmen- tal capacity of pig embryos. However, high or low CO2 levels do not significantly improve the developmental capacity of pig embryos. The best results were obtained for all of the pig embryos at a 5% CO2 concentration.
MDAP-2 is a new antibacterial peptide with a unique structure that was isolated from house- flies. However, its biological characteristics and antibacterial mechanisms against bacteria are still poorly understood. To study the biological characteristics, antibacterial activity, hemolytic activi- ty, cytotoxicity to mammalian cells, and the secondary structure of MDAP-2 were detected; the results showed that MDAP-2 displayed high antibacterial activity against all of the tested Gram-negative bacteria. MDAP-2 had lower hemolytic activity to rabbit red blood cells; only 3.4% hemolytic activity was observed at a concentration of 800μg/ml. MDAP-2 also had lower cytotoxicity to mammalian cells; IC50 values for HEK-293 cells, VERO cells, and IPEC-J2 cells were greater than 1000 μg/ml. The circular dichroism (CD) spectra showed that the peptide most- ly has α-helical properties and some β-fold structure in water and in membrane-like conditions. MDAP-2 is therefore a promising antibacterial agent against Gram-negative bacteria. To deter- mine the antibacterial mechanism(s) of action, fluorescent probes, flow cytometry, and transmis- sion electron microscopy (TEM) were used to study the effects of MDAP-2 on membrane perme- ability, polarization ability, and integrity of Gram-negative bacteria. The results indicated that the peptide caused membrane depolarization, increased membrane permeability, and destroyed membrane integrity. In conclusion, MDAP-2 is a broad-spectrum, lower hemolytic activity, and lower cytotoxicity antibacterial peptide, which is mainly effective on Gram-negative bacteria. It exerts its antimicrobial effects by causing bacterial cytoplasm membrane depolarization, increas- ing cell membrane permeability and disturbing the membrane integrity of Gram-negative bacte- ria. MDAP-2 may offer a new strategy to for defense against Gram-negative bacteria.
Feeder cells can promote cell proliferation and help overcome the developmental arrest of early embryos by producing growth factors. The objective of this study was to evaluate the effects of feeder cells on the development of all single porcine parthenogenetic embryos in vitro. Firstly, we showed that the cleavage and blastocyst formation rate of all single procine parthenogenetic embryos co-cultured with feeder cells increased in contrast to those cultured without feeder cells (p<0.05). However, no statistically significant differences were observed between the blastocyst formation rate in the embryos co-cultured with 3 different kinds feeder cells namely oviduct epithelial feeder cells, granulose feeder cells and porcine fetal fibroblast feeder cells (p>0.05). Secondly, highly significant differences were observed between the cleavage and blastocyst formation rate (p<0.05) when the embryos were co-cultured with oviduct epithelial feeder cells in different volume drops ranging from 3 to 20 μL and the cleavage rate were the highest when cultured in 5 μL drops. Thirdly, the tempospacial pattern of the development of single embryos co-cultured with oviduct epithelial feeder cells was consistent with that of traditional multi-embryo culture, indicating that the co-culturing does not affect the developmental competence of the porcine parthenogenetic embryos. Finally, highly significant differences were observed between the cleavage and blastocyst formation rate with and without zona pellucida in vitro (p<0.05). In this study, a new adaption of in vitro co-culture of single porcine parthenogenetic embryos using feeder cells has been successfully established and this will facilitate further investigations to discover the mechanistic mode of developmental arrest of porcine embryos.
Sapelovirus A (SV-A) is a positive-sense single-stranded RNA virus which is associated with acute diarrhea, pneumonia and reproductive disorders. The virus capsid is composed of four proteins, and the functions of the structural proteins are unclear. In this study, we expressed SV-A structural protein VP1 and studied its antigenicity and immunogenicity. SDS-PAGE analysis revealed that the target gene was expressed at high levels at 0.6 mM concentration of IPTG for 24 h. The mouse polyclonal antibody against SV-A VP1 protein was produced and reached a high antiserum titer (1: 2,048,000). Immunized mice sera with the recombinant SV-A VP1 protein showed specific recognition of purified VP1 protein by western blot assay and could recognize native SV-A VP1 protein in PK-15 cells infected with SV-A by indirect immunofluorescence assay. The successfully purified recombinant protein was able to preserve its antigenic determinants and the generated mouse anti-SV-A VP1 antibodies could recognize native SV-A, which may have the potential to be used to detect SV-A infection in pigs.