NBPF4 mitigates progression in colorectal cancer through the regulation of EZH2-associated ETFA.

Colorectal cancer (CRC) is one of the leading causes of death worldwide, and hence, there is a need to elucidate the molecular mechanisms contributing to the progression of CRC. In this study, we aimed at assessing the role of long non-coding RNA NBPF4 on the tumorigenesis of CRC. Silencing or overexpression experiments were performed on HCT116 and SW260 in vitro models. BALB/c athymic female nude mice aged 5-6 weeks were used as in vivo models. To assess the relationship between NBPF4 and its regulatory RNA pull-down assay, RNA immunoprecipitation, luciferase activity, Western blotting and qRT-PCR were employed. Initially, we identified that NBPF4 was downregulated in CRC tissues and cell lines. Furthermore, we observed that NBPF4 decreased tumorigenesis in both in vitro and in vivo models. Additionally, we identified that ETFA was highly expressed in CRCs and was negatively associated with NBPF4. Subsequently, we identified that EZH2, a transcriptional factor, activated ETFA by enhancing the methylation of its promoter, and EZH2 was also highly regulated in CRCs. Using COAD and READ databases, we confirmed that EZH2 and ETFA were positively correlated. Furthermore, we identified NBPF4 and EZH2 were targets for ZFP36, which bound and positively regulated NBPF4. This prevented NBPF4 from binding to its negative regulator miR-17-3p. Our results demonstrated that NBPF4 downregulated EZH2 and stabilized itself by binding to ZFP36, thus escaping from inhibition by miR-17-3p, which allowed mitigation of CRC through inhibition of ETFA.

[Study on seeds after-ripening regulation and key techniques for seedling of Pulsatilla cernua].

OBJECTIVE: The seeds of Pulsatilla cernua were used as tested materials for screening and establishing the main factors with different levels to control fast development of seed embryonic and seedlings of Pulsatilla cernua. METHODS: The main factors with different levels for development of seed embryonic and seedlings of Pulsatilla cernua were investigated through repeated experiments with multifactorial and cross. RESULTS: The method for development of seedlings and seeds germination of Pulsatilla cernua were to soak the seeds in the mixed solutions with 2.40 mg/L KT, 2.80 mg/L GA3 and 0.30 -0.70 mg/L 2,4-D for 24 h. The seeds and sand (1:2) were mixed, treated with temperature change in 63 - 70 d. The extent of temperature change and time were (23 ± 2) degrees C and 14 h in day, while (10 ± 2) degrees C and 10 h in night. The incidence rate of the embryo with cotyledons was 95.1%, and the germination rate of seed was 92.3%. CONCLUSION: The plant regeneration control technology for development of seed embryonic and seedling of Pulsatilla cernua have been solved, which is suitable for industrial seedlings of Pulsatilla cernua.

Messenger RNA profiling for forensic body fluid identification: research and applications.

Identifying the origin of body fluids left at a crime scene can give a significant insight into crime scene reconstruction by supporting a link between sample donors and actual criminal acts. However, the conventional body fluid identification methods are prone to various limitations, such as time consumption, intensive labor, nonparallel manner, varying degrees of sensitivity and limited specificity. Recently, the analysis of cell-specific messenger RNA expression (mRNA profiling) has been proposed to supplant conventional methods for body fluid identification. Since 2011, the collaborative exercises have been organized by the European DNA Profiling Group (EDNAP) in order to evaluate the robustness and reproducibility of mRNA profiling for body fluid identification. The major advantages of mRNA profiling, compared to the conventional methods, include higher sensitivity, greater specificity, the ability of detecting several body fluids in one multiplex reaction, and compatibility with current DNA extraction and analysis procedure. In the current review, we provided an overview of the present knowledge and detection methodologies of mRNA profiling for forensic body fluid identification and discussed its possible practical application to forensic casework.

Andrographolide promoted ferroptosis to repress the development of non-small cell lung cancer through activation of the mitochondrial dysfunction.

BACKGROUND: Ferroptosis, a form of regulated cell death by lipid peroxidation, was currently considered as a key factor affecting the occurrence and progression in various cancers. Andrographolide (ADE), a major effective ingredient of Andrographis paniculate, has proven to have a substantial anti-tumor effect on multiple cancer types. However, the function and underlying mechanism of ADE in Non-Small Cell Lung Cancer remain unclear. METHODS: CCK8 assay, colony-formation assay, flow cytometry, scratch test, transwell assay, western blotting, ferroptosis analysis and mitochondria analysis were performed to reveal the role and underlying mechanisms of ADE in NSCLC cell lines (H460 and H1650). In vivo, xenograft model and lung metastatic model were performed to verify the effect of ADE on the growth and metastasis of NSCLC. RESULTS: In this present study, we demonstrated that treatment with ADE could inhibit cell growth and metastases through eliciting ferroptosis in vitro an in vivo. The IC50 of ADE in H460 and H1650 cells were 33.16 µM and 32.45 µM respectively. In Lewis xenografted animals, i.p. ADE repressed relative tumor growth (p < 0.01) and inhibited metastases (p < 0.01). Notably, the ferroptosis inhibitor Fer-1 abrogated the anti-tumor capacity of ADE. Induction of ferroptosis by ADE was confirmed by elevated levels of reactive oxygen sepsis (ROS), glutathione (GSH), malondialdehyde (MDA), intracellular iron content and lipid ROS reduced glutathione (GSH) accumulation (p < 0.01). Furthermore, ADE inhibited the expression of ferroptosis-related protein GPX4 and SLC7A11. Simultaneously, it also disclosed that ADE enhanced mitochondrial dysfunction, as evidenced by increased mitochondrial ROS release, mitochondrial membrane potential (MMP) depolarization, and decreased mitochondrial ATP. Most interestingly, Mito-TEMPO, a mitochondria-targeted antioxidant, rescued ADE-induced ferroptosis. CONCLUSION: Our data validated that ADE treatment could restrain proliferation and metastases of NSCLC cells through induction of ferroptosis via potentiating mitochondrial dysfunction.

Evaluating the Effect of Forage Rape (Brassica napus) Ensiling Kinetics on Degradability and Milk Performance as Non-conventional Forage for Dairy Buffalo.

The recent increase in demand for animal protein sources has led to the urgency to introduce non-conventional feed sources and opened the space to study feed management and its effects on animal productivity. Forage rape (Brassica napus L.) is a high-quality forage crop with a remarkable nutritional value and productive and fast growth capacity; however, studies on processing methods are limited. This study evaluates the effect of an ensiling process on rape silage quality kinetics, in situ degradability, and milk responses in dairy buffaloes. Firstly, the whole-plant forage rape was ensiled, and silage samples were collected 30, 60, and 90 days after ensiling to determine pH, evaluation of sensory characteristics, and chemical composition. Then, samples were taken for further chemical analysis at days 30, 60, and 90. After that, the degradability of the dry matter (DM) and crude protein (CP) of the silage was evaluated by an in situ degradability experiment using three fistulated buffalos (550 ± 20 kg body weight, 4.7 ± 0.76 years). Finally, whole-plant rape silage (after 60 days) was included in a 10, 20, and 30% of DM dairy buffalo diet in the lactating buffalo ration. The results showed that silage pH did not change significantly during the ensiling process (p > 0.05); however, the silage achieved the optimal comprehensive sensory characteristic score from days 30 to 60. There was also a significant change in neutral detergent fiber (NDF) content and acid detergent fiber content, which decreased significantly (p = 0.001 and p < 0.001, respectively). Ensiling of the whole-plant rape significantly reduced effective DM degradability (p < 0.05) without altering CP degradability (p > 0.05). Furthermore, the inclusion of forage rape silage linearly (p = 0.03) increased milk fat and protein contents and did not affect milk yield, lactose, and urea nitrogen contents in raw buffalo milk. In conclusion, whole-plant rape silage could significantly maintain the optimal ether extract (EE) protein content without affecting CP degradability, in addition to improving milk fat and milk protein. Therefore, ensiling may be an efficient method of forage rape utilization, and forage rape silage can be recommended as a good forage source for dairy buffaloes.

Cu2O hollow spheres: synthesis, characterization and magnetic property.

Cu2O hollow spheres with thin shells were synthesized using poly(vinylpyrrolidone) (PVP) as the template in deionized water. In this process, CuAc2.H2O was reduced by acerbic acid in the presence of PVP and sodium hydroxide. Techniques, including XRD, XPS, TEM and HRTEM were used to characterize the morphology, structure and composition of the as-prepared nanomaterials. The results revealed that the average diameter of polycrystalline Cu2O hollow spheres is about 200 nm, with the shells as thin as approximately 10 nm. The formation mechanism, of these hollow spheres was studied. Magnetic property of the sample demonstrates a paramagnetic behavior at 15 K, and it might be attributed to the hollow-sphere structure of the diamagnetic Cu2O.

[Analysis of projects of schistosomiasis sponsored by National Science Foundation of China].

OBJECTIVE: To summarize the present development by analysis of projects in schistosomiasis funded by National Science Foundation of China (NSFC). METHODS: Based on the ISIS database of NFSC, the projects in the studies of schistosomiasis from 2005 to 2016 were analyzed. The distributions of sponsored numbers, amounts, types, agencies, disciplines and changes in research topics by means of network profiles were described. RESULTS: During the study period, 198 projects were funded by NSFC totally with 76.05 million yuan in which the general and youth projects were main types. The main sponsored agencies were research institutes and medical colleges. The top three fields sponsored were medical pathogenic microbes and infection, veterinary and medical immunology. CONCLUSIONS: The funding on schistosomiasis researches has a downward trend, but studies are continuing in depth. In this situation, innovative and interdisciplinary researches need to be encouraged to promote the development of schistosomiasis.

Novel ultra-low temperature co-fired microwave dielectric ceramic at 400 degrees and its chemical compatibility with base metal.

A novel NaAgMoO4 material with spinel-like structure was synthesized by using the solid state reaction method and the ceramic sample was well densified at an extreme low sintering temperature about 400°C. Rietveld refinement of the crystal structure was performed using FULLPROF program and the cell parameters are a = b = c = 9.22039 Å with a space group F D -3 M (227). High performance microwave dielectric properties, with a permittivity ~7.9, a Qf value ~33,000 GHz and a temperature coefficient of resonant frequency ~-120 ppm/°C, were obtained. From X-ray diffraction (XRD) and Energy Dispersive Spectrometer (EDS) analysis of the co-fired sample, it was found that the NaAgMoO4 ceramic is chemically compatible with both silver and aluminum at the sintering temperature and this makes it a promising candidate for the ultra-low temperature co-fired ceramics technology. Analysis of infrared and THz spectra indicated that dielectric polarizability at microwave region of the NaAgMoO4 ceramic was equally contributed by ionic displasive and electronic polarizations. Its small microwave dielectric permittivity can also be explained well by the Shannon's additive rule.

Early IL-4 production does not predict susceptibility to Leishmania major.

Leishmania major in mice can be self-limiting or fatal, depending upon the inbred mouse strain. It is well established that the outcome of infection is dependent upon the Th cell subset that dominates after infection. This has led to intense study of the early events associated with infection, in order to better understand the factors that determine Th1/2 cell development. In the present report, we have analyzed the kinetics of IL-4 and IL-4 mRNA production in three mouse strains: BALB/c, C3H, and C57BL/6. We found that in the first week IL-4 is absent in the C3H mice, but present in the susceptible BALB/c and relatively resistant C57BL/6 mouse. These data indicate that the presence of IL-4 by itself does not determine whether the immune response will be dominated by Th2 cells, since C57BL/6 mice will eventually develop a Th1 response and heal. We suggest that the critical cytokine that determines susceptibility in experimental leishmaniasis is IL-12, rather than IL-4. Thus, in C3H mice IL-12 is evident soon after infection, and IL-4 responses are not observed. In C57BL/6 mice, IL-12 production is delayed, but once evident, the IL-4 response is ablated. Further, we show that addition of IL-12 can block early IL-4 production in BALB/c mice, and neutralization of IL-12 in C3H mice uncovers IL-4 production in response to L. major infection. Taken together, these data indicate that susceptibility to L. major, while possibly requiring IL-4, is not determined by the presence or absence of IL-4.