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The key to successful in vitro embryo production is to mimic the natural in vivo oviductal microenvironment. Although the chemically defined media in extensive use for the in vitro culture of mammalian embryos is based on the composition of oviductal fluid, the in vitro embryo production (IVEP) systems in current use must still bypass the oviduct to produce embryos in vitro. Extracellular vesicles (EVs) in the oviduct are versatile intercellular delivery vehicles for maternal-embryo communication, and a lack of them can be associated with failed early embryonic development under in vitro culture conditions. Herein, we isolated EVs from porcine oviduct fluid and confirmed that oviductal EV supplementation improves the embryonic development of parthenogenetically activated (PA) embryos in terms of blastocyst formation rates and total cell numbers. Our experiments also revealed that a beneficial effect of oviductal EVs on PA embryos was achievable, at least in part, by relieving endoplasmic reticulum stress. These results suggest that the maternal-embryo communication mediated by oviductal EVs benefits early embryonic development. Given the contribution of oviductal EVs to early embryonic development, these findings offer novel insights for the optimization of current IVEP systems. This article is protected by copyright. All rights reserved.

γ-Oryzanol, which is rich in rice seeds, is natural phytonutrition with many functions such as antioxidant. However, γ-oryzanol has a large loss in the traditional processing, resulting in a huge waste of available resources. In this study, the rice seeds highest content of γ-oryzanol during the development of rice seeds was selected and separated processes by nanofiltration. Several nanofiltration membranes were dramatically applied to condensing phytonutrition under 0.5 Mpa pressure and the flux ranged from 5.8 to 7.7 kg•m-2•h-1. The first processing supplied the separation improving ethanol-extraction system enrichment in γ-oryzanol. In the second, refining ethanol-extraction dramatically reduced the content of free fatty acids, while γ-oryzanol in the retention solution was further significantly increased. Moreover, the isolated γ-oryzanol added in the soybean oil, notably improved antioxidant ability of oil. Our founding can be widely used in the functional food manufacturing and also provides as an available method.

B cell lymphoma-2-like 2 (BCL2L2), an important regulator of apoptosis, plays vital roles in several physiological processes, as revealed by studies in humans and mice. However, reports on pig BCL2L2 are few, and the encoding gene has not been identified experimentally. This study was designed to clone the porcine BCL2L2 gene and its alternative splicing (AS) transcripts using molecular biological techniques and to analyze the regulatory mechanisms underlying transcription and translation. The BCL2L2 cDNA (V1) was 807 bp in length and encoded a polypeptide of 193 aa containing four BCL-2 homology domains. A total of nine AS transcripts were obtained, among which V2 and V3 differed from V1 in the 5' untranslated region (UTR). The core promoter was mapped to a range of -1102 to -759 bp (the first nucleotide of the start codon was designated as +1). There were several functional cis-elements, including one SP1 and two C/EBPα binding sites at around -759 bp. AS in the 5' UTR is involved in the regulation of gene expression, as revealed by dual-luciferase reporter and western blot analysis, and the secondary structure of the 5' UTRs may be the reason for the differential expression of V1-3. At the same time, an upstream open reading frame (ORF) existed in each of the three 5' UTRs, was found to repress the expression of the main ORF. Additionally, the roles of porcine BCL2L2 in cell proliferation and apoptosis were preliminarily analyzed. The results will contribute to further characterizing the role of BCL2L2.

Two novel compounds, 2-(2-hydroxyethylthio)-5,8-dimethoxy-1,4-naphthoquinone (HEDMNQ) and 2-(6-hydroxyhexylthio)-5,8-dimethoxy-1,4-naphthoquinone (HHDMNQ), were synthesized to investigate the kill effects and mechanism of 1,4-naphthoquinone derivatives in lung cancer cells. The results of the CCK-8 assay showed that HEDMNQ and HHDMNQ had significant cytotoxic effects on A549, NCI-H23, and NCI-H460 NSCLC cells. Flow cytometry and western blot results indicated that HHDMNQ induced A549 cell cycle arrest at the G2/M phase by decreasing the expression levels of cyclin-dependent kinase 1/2 and cyclin B1. Fluorescence microscopy and flow cytometry results indicated that HHDMNQ could induce A549 cell apoptosis, and western blot analysis showed that HHDMNQ induced apoptosis through regulating the mitochondria pathway, as well as the MAPK, STAT3, and NF-κB signalling pathways. Flow cytometry results showed that intracellular reactive oxygen species (ROS) levels were increased after HHDMNQ treatment, and western blot showed that ROS could modulate the intrinsic pathway and MAPK, STAT3, and NF-κB signalling pathways. These effects were blocked by the ROS inhibitor N-acetyl-L-cysteine in A549 cells. Our findings suggest that compared with HEDMNQ, HHDMNQ had the stronger ability to inhibit the cell viability of lung cancer cells and induce apoptosis by regulating the ROS-mediated intrinsic pathway and MAPK/STAT3/NF-κB signalling pathways. Thus, HHDMNQ might be a potential antitumour compound for treating lung cancer.

We compared the ecological characteristics of tussock individuals and populations undergoing natural and artificial restoration in Carex tussock wetlands in the Sun Island in Harbin and identified the relationships between the growth of Carex tussock and environmental factors. Results showed that there were obvious seasonal dynamics in morphological characteristics of C. appendiculata. Tussocks grew rapidly from May to June, peaked in June, and then decreased steadily from July to August. There were significant differences in ecological characteristics of Carex tussocks between natural and artificial restorations. The morphological characteristics of individual tussock, including leaf area, leaf width, fresh weight per ramet, dry weight per ramet, and the hummock shape indicators (hummock height, diameter, volume and surface area) in natural restored area were significantly higher than those in artificial transplanting area. For the Carex tussock community, tussock density, coverage and biomass in natural restoration area were significantly lower than those in artificial transplanting area. Soil water content, water depth and hummock spacing in natural restoration area were significantly higher than those in artificial restoration area, which facilitated the formation and development of individual tussock. Higher transplanting density was the main factor leading to higher density, coverage, and biomass in artificial restoration area. Our results suggested that the distribution characteristics of tussocks in natural restoration area should be taken into account in future restoration and protection works. Appropriate adjustment of the distance between hummock (54.22-117.89 cm) and population density (1.9-3.1 ind·m本研究以哈尔滨太阳岛草丘湿地为对象,对比了人工恢复与自然恢复下苔草草丘个体和种群的生态特征,并分析其与环境因子的关系.结果表明:苔草植株生长随时间呈现先增加后降低的变化趋势(5—8月),初期(5—6月)生长迅速,6月达到峰值.人工恢复和自然恢复模式下,苔草草丘个体和种群特征差异显著:自然恢复下苔草叶面积、叶宽、单株鲜重、单株干重、丘墩高度、直径、丘顶面积、丘墩表面积、丘墩体积等苔草草丘个体特征均显著高于人工恢复,人工恢复下苔草草丘密度、盖度、生物量等种群特征显著高于自然恢复,物种多样性无显著差异.土壤含水量、水深、草丘密度、丘间距离是导致2种恢复模式下苔草草丘生长差异的主要因素,自然恢复区土壤含水量、水深、间距均显著高于人工恢复区,对草丘个体的形成和发育具有促进作用,人工恢复区高移栽密度是导致草丘密度、盖度、生物量高于自然恢复区的主要因素.建议未来开展苔草草丘湿地恢复和保护时,应参考自然恢复湿地中草丘的分布特点,适当调整丘间距离(54.22～117.89 cm)和种群密度(1.9～3.1 墩·m

Min pigs and Large White pigs are two breeds that vary significantly in thickness of adipose tissue. In this study, transcriptome profiling was used to examine the fat tissues of both breed...

The transcription factor forkhead box N1 (Foxn1) plays an important role in the development and function of thymic epithelial cells (TECs) in vertebrates. However, the transcriptional regulation of Foxn1 is still unknown. A series of dual luciferase report vectors were constructed and their relative activities were also detected. The 5'-untranslated regions contains two cis-acting elements, Sp1 and GATA-1, as well as trans-acting elements between positions -332 to -438. Nevertheless, the relative activities of Foxn1 promoter were significantly increased in PK15 cells only when the Sp1 was overexpressed, suggesting that the Sp1 was the most important sequence for Foxn1 transcription activation in the pig. Regrettably, the exact trans-acting elements were not found.

The effect of genistein on Bcl-2 and Bax protein expression in the ovarian tissue of rats with polycystic ovarian syndrome (PCOS) was evaluated. Sixty rats were divided into six groups. Rats in the Dose group received genistein at a concentration of either 5 (L-gen), 10 (M-Gen) or 20 (H-Gen) mg per kg of body weight per day. The expression of Bcl-2 mRNA and Bax mRNA was determined by in situ hybridization. Bcl-2 and Bax protein concentration was quantified by ELISA. The results showed that the expression of Bcl-2 mRNA and Bcl-2 protein was significantly higher in the high genistein Dose group (H-Gen) when compared to the Model group (MG) (P

// Ying Wang 1, 4 , Xue Han 2 , Yan Dong Li 3 , Yabing Wang 2 , Shi Yang Zhao 2 , Dong Jie Zhang 1 and Yu Lu 5 1 College of Food, Heilongjiang Bayi Agricultural University, Daqing 163319, PR China 2 College of Biological Science and Engineering, Hebei University of Science and Technology, Shijiazhuang 050018, PR China 3 Hebei Institute of Veterinary Drugs Control, Shijiazhuang 050000, PR China 4 National Coarse Cereals Engineering Research Center, Daqing 163319, PR China 5 Huabei Petroleum Administration Bureau, Huasheng Integrated Service, Tianjin 300000, PR China Correspondence to: Xue Han, email: hanxue_h@163.com Keywords: lentinan; dose dependence; liver cancer; immunoprophylaxis; promotion Received: March 12, 2017 Accepted: April 21, 2017 Published: August 01, 2017 ABSTRACT Lentinan could exhibit significant biological activity favorable for human health and disease control such as the recovery of patients with liver cancer. In order to investigate the effect of lentinan dose dependence between immunoprophylaxis and promotion of cancer cell proliferation of the murine liver cancer, different concentrations of lentinan were prepared for the test in vitro (MTT assay) and in vivo (cumulative survival assay, spleen lymphocyte proliferation tests and peritoneal macrophage phagocytosis assays). New emerging proteins of the H22 cell incubated with lentinan was demonstrated by MS analysis and protein database searching. Lentinan was non-toxic for HL7702 cells but inhibited H22 cells proliferation obviously in a dose-dependent manner. In vivo , the proliferation of H22 hepatocarcinoma cells was inhibited by lentinan 0.4mg/kg body weight (L2, survival rate, 20%, P

Min Pig (Sus scrofa) is a northern Chinese domestic pig breed, whose cold resistance is remarkable. However, the cold-resistant phenotype of Min Pig is unclear. Our studies showed that in long-term...

Oocyte competence is an important determinant of reproductive success and can be disrupted by heat/cold stress. Information related to seasonally induced cold stress effects in cold areas on oocyte competence in the bovine is still scarce, however. Therefore, we performed experiments to test the hypothesis that seasonally induced cold/heat stress affects bovine oocyte competence in cold areas. Oocytes were collected in the winter, summer, and autumn, and we showed a significantly better survival and growth rate in the cold compared with hot temperatures, as recorded for two-cell and blastocyst stage embryos (P

Myogenin (MyoG) plays an important role in muscle differentiation, which can initiate the formation of the fusion of muscle cells and myofibroblasts. To investigate the transcriptional regulatory mechanism of MyoG gene in Min pig, the promoter fragment spanning from −2085 bp to +118 bp was amplified with PCR, and the potential transcriptional elements were predicted via bioinformatics analysis. Subsequently, nine promoter segments with different base pair sizes were obtained and ligated into PGL3-Basic vectors, respectively. Lastly, the functional activity of each of the aforementioned promoters was tested separately in transiently transfected C2C12 cells via a dual-luciferase assay system. The results indicated that the sequence −199 bp to −31 bp may have the basic activity of the promoter. Further analysis demonstrated that the sequence −137 bp to −101 bp was important for the basic transcriptional activity. Bioinformatics analysis showed that CdxA and MZF1 transcription factors may play an impo...

The relationship between species richness and elevation is a hot issue in ecology and has been documented extensively. It is widely accepted that area size can significantly affect this relationship and thus mask the effects of other predictors. Despite the importance of the relationship between species richness and elevation while accounting for the area effect, it is insufficiently studied. Here, we evaluated area-corrected species richness patterns of all vascular plants as well as six vascular plant subgroups (seed plants, ferns, trees, shrubs, herbs and vines) along a tropical elevational gradient (Hainan Island, China). If assessed in equal-elevation bands, uncorrected species richness showed bell-shaped curves, while area-corrected species richness assessed in equal-area bands appeared to increase monotonically due to the small proportion of highlands on Hainan Island. The mid-domain effect (MDE) was significantly correlated with both uncorrected and area-corrected species richness. On Hainan Island, the use of equal-area elevational bands increased the explanatory power of MDE. These findings provide useful insights to adjust for the area effect and highligh t the need to use equalarea bands along the elevational gradient.

Isoorientin (ISO) is a naturally occurring C‑glycosyl flavone that has various pharmacological properties, such as anti‑bacterial and anti‑inflammatory effects. However, its underlying molecular mechanisms in human lung cancer cells remain unknown. In the present study, the effects of ISO on the induction of apoptosis and relative molecular mechanisms in A549 human lung cancer cells were investigated. The results of Cell Counting Kit‑8 assay (CCK‑8) indicated that ISO exerted significant cytotoxic effects on 3 lung cancer cell lines, but had no obvious side‑effects on normal cells. Moreover, flow cytometry and western blot analysis revealed that ISO induced mitochondrial‑dependent apoptosis by reducing mitochondrial membrane potential. ISO also increased the expression levels of Bax, cleaved‑caspase‑3 (cle‑cas‑3) and poly(ADP‑ribose) polymerase (PARP; cle‑PARP), and decreased the expression levels of Bcl‑2 in A549 cells. Furthermore, ISO induced G2/M cell cycle arrest by decreasing the expression levels of cyclin B1 and CDK1/2, and increasing the expression levels of p21 and p27 in A549 cells. As the duration of ISO treatment increased, intracellular reactive oxygen species (ROS) levels in A549 cells also increased. However, pre‑treatment of the cells with the ROS scavenger, N‑acetylcysteine (NAC), inhibited ISO‑induced apoptosis. In addition, ISO increased the expression levels of p‑p38, p‑JNK and IκB‑α; and decreased the expression levels of p‑extracellular signal‑regulated kinase (ERK), p‑signal transducer and activator of transcription (STAT)3, p‑nuclear factor (NF)‑κB, NF‑κB and p‑IκB; these effects were induced by mitogen‑activated protein kinase (MAPK) inhibitors and blocked by NAC. Taken together, the results of the present study indicate that ISO induces the apoptosis of A549 lung cancer cells via the ROS‑mediated MAPK/STAT3/NF‑κB signaling pathway, and thus may be a potential drug for use in the treatment of lung cancer.

The present study investigated the mechanisms of apoptosis induced by cryptotanshinone (CT) in human rheumatoid arthritis fibroblast‑like synoviocytes (RA‑FLSs). Cell Counting kit‑8 assay was performed to determine the cytotoxic effects of CT in human RA‑FLSs, including primary RA‑FLS, HFLS‑RA and MH7A cells, and in HFLS cells derived from normal synovial tissue. Annexin V‑FITC/PI staining was used to detect the apoptotic effects of CT in HFLS‑RA and MH7A cells. Flow cytometry was performed to detect the apoptotic and reactive oxygen species (ROS) levels induced by CT in HFLS‑RA cells. Western blotting was used to assess the expression levels of proteins associated with apoptosis and with the mitogen‑activated protein kinase (MAPK), protein kinase B (Akt), and signal transducer and activator of transcription‑3 (STAT3) signaling pathways. The results demonstrated that CT treatment significantly suppressed HFLS‑RA and MH7A cell growth, whereas no clear inhibitory effect was observed in normal HFLS cells. CT exposure downregulated the expression levels of B‑cell lymphoma 2 (Bcl‑2), p‑Akt, p‑extracellular signal‑related kinase and p‑STAT3, while it upregulated the expression levels of Bcl‑2‑associated death promoter (Bad), caspase‑3, poly (ADP‑ribose) polymerase (PARP), p‑p38 and p‑c‑Jun N‑terminal kinase. Following ROS scavenging, the CT‑induced apoptosis and altered expression levels of Bcl‑2, Bad, cleaved caspase‑3 and cleaved PARP were restored. Furthermore, the Akt, MAPK and STAT3 signaling pathways were regulated by intracellular ROS. These results suggest that ROS‑mediated Akt, MAPK and STAT3 signaling pathways serve important roles in the CT‑induced apoptosis of RA‑FLSs.

In this study, we examined the association between soy isoflavones and lipid profiles, apolipoprotein levels in patients with type 2 diabetes in China. The study population was composed of 120 cases (80 women with type 2 diabetes and 40 healthy women). Objects in treatment group received isoflavones 435 mg/day for 2 months, then lipid profiles were analyzed by the colorimetry method and apolipoprotein levels were determined by immune turbidimetric method. And all the indexes were determined after oral glucose tolerance test. The levels of total cholesterol, triglyceride and LDL-C significantly reduced and the levels of HDL-C and apolipoprotein A1 significantly raised in the treatment group after intervention (p

Kaempferide is an O-methylated flavonol that has received much attention due to its various biological activities. In this study, we explored the underlying mechanisms of kaempferide in human lung cancer A549 cells. The Cell Counting Kit-8 (CCK-8) assay, Hoechst 33342/propidium iodide double staining, flow cytometry, scratch wound healing assay, and Western blot analysis were used to measure cell apoptosis, the cell cycle, reactive oxygen species (ROS) levels, and cell migration of human lung cancer cells. Kaempferide significantly inhibited human lung cancer cell proliferation, and its toxic effects on normal cells were significantly lower than those of 5-fluorouracil. Kaempferide induced A549 cell apoptosis by decreasing the mitochondrial membrane potential and the expression level of B-cell lymphoma 2, and by increasing the expression levels of Bcl-2-associated X protein and caspase-3. It also regulated mitogen-activated protein kinase (MAPK), signal transducer and activator of transcription 3 (STAT3), and nuclear factor kappa B (NF-κB) signaling pathways by increasing the expression levels of phosphorylated c-Jun N-terminal kinase, p-p38, I kappa B, and by decreasing the expression levels of phosphorylated extracellular signal-regulated kinase, p-STAT3, and NF-κB. Kaempferide induced cell cycle arrest in the G0/G1 phase in A549 cells by downregulating the expression levels of p-AKT, cyclin D1, and cyclin-dependent kinase 2. Furthermore, kaempferide blocked A549 cell migration by downregulating the expression levels of transforming growth factor beta 1 (TGF-β1), p-β-catenin, p-glycogen synthase kinase 3 beta, N-cadherin, and vimentin, and by upregulating the expression level of E-cadherin. Kaempferide enhanced the accumulation of ROS, and N-acetyl-l-cysteine (a ROS inhibitor) decreased the regulation of MAPK, NF-κB, AKT, and TGF-β signaling pathways by kaempferide, inhibited cell apoptosis, and reversed cell cycle arrest. Our results showed that kaempferide induced apoptosis via ROS-mediated MAPK, NF-κB, AKT, and TGF-β signaling pathways in A549 cells. Thus, kaempferide may be a novel drug candidate for lung cancer.

Derivatives of 1,4‑naphthoquinone have excellent anti‑cancer effects, but their use has been greatly limited due to their serious side effects. To develop compounds with decreased side effects and improved anti‑cancer activity, two novel types of 1,4‑naphthoquinone derivatives, 2,3‑dihydro‑2,3‑epoxy‑2‑propylsulfonyl‑5,8‑dimethoxy‑1,4‑naphthoquinone (EPDMNQ) and 2,3‑dihydro‑2,3‑epoxy‑2‑nonylsulfonyl‑5,8‑dimethoxy‑1,4‑naphthoquinone (ENDMNQ) were synthesized and their anti‑tumor activities were investigated. The effects of EPDMNQ and ENDMNQ on cell viability, apoptosis and accumulation of reactive oxygen species (ROS) in liver cancer cells were determined by MTT cell viability assay and flow cytometry. The expression levels of mitochondrial, mitogen activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3) signaling pathway‑associated proteins in Hep3B liver cancer cells were analyzed by western blot analysis. The results demonstrated that EPDMNQ and ENDMNQ inhibited the proliferation of liver cancer Hep3B, HepG2, and Huh7 cell lines but not that of normal liver L‑02, normal lung IMR‑90 and stomach GES‑1 cell lines. The number of apoptotic cells and ROS levels were significantly increased following treatment with EPDMNQ and ENDMNQ, and these effects were blocked by the ROS inhibitor N‑acetyl‑L‑cysteine (NAC) in Hep3B cells. EPDMNQ and ENDMNQ induced apoptosis by upregulating the protein expression of p38 MAPK and c‑Jun N‑terminal kinase and downregulating extracellular signal‑regulated kinase and STAT3; these effects were inhibited by NAC. The results of the present study demonstrated that EPDMNQ and ENDMNQ induced apoptosis through ROS‑modulated MAPK and STAT3 signaling pathways in Hep3B cells. Therefore, these novel 1,4‑naphthoquinone derivatives may be useful as anticancer agents for the treatment of liver cancer.

Follistatin (FS) is a secreted protein, which was originally isolated from porcine follicular fluid. Expression of follistatin is tightly regulated during porcine growth and development. To study the essential transcriptional regions of the porcine FS promoter, ten primer pairs were designed to amplify segments with different lengths of the FS promoter from −1 800 to +16 bp. The products were then inserted into the pGL3-basic vector to analyze the relative luciferase activity. The results showed that the most remarkable changes of promoter activity were observed between constructs (−302/+16 bp)-FS and (−180/+16 bp)-FS (P

Lumbar disc herniation (LDH) is the most common cause of radiculopathy, whose pathological entity underlying nerve root compression is usually on the same side as the symptoms. However, LDH causing contralateral radiculopathy are sometimes encountered by pain physicians. There have been tremendous developmens in the treatment options for LDH; the situation of LDH causing contralateral radiculopathy is indeed a dilemma for some pain physicians. We will report a case of a patient with a L4-5 disc herniation whose left herniated disc caused radiculopathy on the right side. After a percutaneous lumbar endoscopic discectomy via the side ipsilateral to the symptomatic side, this case obtained a significant symptom remission. The migrated epidural fat is discussed as a cause of associated contralateral neurological deficit. Only via a surgical approach ipsilateral to the herniated side, could there be a clinical improvement postoperatively. Key words: Contralateral symptoms, lumbar disc herniation, radiculopathy, epidural fat

Background context Percutaneous vertebroplasty (PVP) has proven to be a valuable palliative treatment option for patients with medically refractory painful osteolytic metastases of the spine. Percutaneous vertebroplasty of the atlas has been reported in only seven articles and has been performed with different techniques and approaches. Purpose To describe the technique we used to perform PVP of a lytic lesion of the lateral mass of C1 via anterior retropharyngeal approach guided by C-arm fluoroscopy. Study design A technical report. Patient sample It included a 75-year-old man with known metastatic lung carcinoma and incapacitating right suboccipital and neck pain refractory to conventional medical treatment. Radiologic evaluation showed revealed osteolytic destruction of C1 and C2, mainly invading the right lateral mass of C1 and the vertebral body of C2. Outcome measures The right suboccipital and neck pain was measured using the visual analog scale (VAS). Methods Under C-arm fluoroscopy, a novel anterior retropharyngeal approach, through the vertebral body of C2 into the metastatic osteolytic vertebral lesion of C1, was performed to achieve the PVP in C1 followed by a PVP in C2. Results Immediately after the operation, the patient reported substantial pain relief (from VAS 9/10 preoperatively to 3/10). At 12 hours postoperatively, the range of motion was also improved. There were no surgery-related complications. The immediately postoperative cervical plain film and computed tomography scan showed adequate filling of the osteolytic lesion without the obvious leakage of bone cement. Clinical follow-up at 3 months revealed that this pain condition was improved and maintained (VAS 1/10). Conclusions When the transoral approach is unsuitable or contraindicated, the anterior retropharyngeal approach could be an efficacious alternative in selected patients with C1 metastasis, providing adequate filling of bone cement and significant pain relief. Based on our preliminary exploration, only assisted by C-arm fluoroscopy, this approach is feasible to achieve PVP in C1 under local anesthesia and intravenous analgesia. Nevertheless, when considering the substantial potential risks, this technically challenging procedure should be performed by experienced operators.