Circ_0001714 knockdown alleviates lipopolysaccharide-induced apoptosis and inflammation in renal tubular epithelial cells via miR-129-5p/TRAF6 axis in septic acute kidney injury.

BACKGROUND: Circular RNAs (circRNAs) have been shown to play roles in regulating sepsis. Sepsis is a major cause of acute kidney injury (AKI). Herein, we aimed to investigate the role and mechanism of circ_0001714 in the progression of sepsis-induced AKI. METHODS: Human HK-2 cells were exposed to lipopolysaccharide (LPS) for functional experiments. Quantitative real-time polymerase chain reaction and western blotting were used for expression analysis. Functional experiments were performed by using MTT assay, 5-ethynyl-2'-deoxyuridine assay, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). The binding between miR-129-5p and circ_0001714 or TRAF6 (TNF receptor associated factor 6) was validated using dual-luciferase reporter assay. RESULTS: Circ_0001714 expression was higher in sepsis-AKI patients. HK-2 cells were exposed to LPS to imitate the injury of renal tubular epithelial cells during sepsis-AKI. LPS dose-dependently up-regulated circ_0001714, moreover, circ_0001714 silencing reversed LPS-evoked apoptosis and inflammation in HK-2 cells. Mechanistically, circ_0001714 sequestered miR-129-5p to up-regulate TRAF6 expression, implying the circ_0001714/miR-129-5p/TRAF6 feedback loop. MiR-129-5p was decreased, while TRAF6 was increased in sepsis-AKI patients and LPS-stimulated HK-2 cells. MiR-129-5p re-expression or TRAF6 silencing protected against LPS-induced HK-2 cell apoptosis and inflammation. Additionally, a series of rescue experiments showed that miR-129-5p inhibition reversed the inhibitory action of circ_0001714 knockdown on LPS-induced HK-2 cell injury. Furthermore, TRAF6 overexpression also attenuated the protective effects of miR-129-5p on HK-2 cells under LPS treatment. CONCLUSION: Circ_0001714 silencing might alleviate LPS-induced apoptosis and inflammation via targeting miR-129-5p/TRAF6 axis in HK-2 cells.

Exploring the limits of virtual source localization with amplitude panning on a flat panel with actuator array: Implications for future researcha).

Immersive and spatial sound reproduction has been widely studied using loudspeaker arrays. However, flat-panel loudspeakers that utilize thin flat panels with force actuators are a promising alternative to traditional coaxial loudspeakers for practical applications, with benefits in low-visual profiles and diffuse radiation. Literature has addressed the sound quality and applications of flat-panel loudspeakers in three-dimensional sound reproduction, such as wave field synthesis and sound zones. This paper revisits the spatial sound perception of flat-panel loudspeakers, specifically the localization mismatch between the perceived and desired sound directions when using amplitude panning. Subjective tests in an anechoic chamber with 24 subjects result in the mean azimuth direction mismatch within ±6.0° and the mean elevation mismatch within ±10.0°. The experimental results show that the virtual source created by amplitude panning over a flat-panel loudspeaker still achieves spatial localization accuracy close to that of a real sound source, despite not using complex algorithms or acoustic transfer function information. The findings of this study establish a benchmark for virtual source localization in spatial sound reproduction using flat-panel loudspeakers, which can serve as a starting point for future research and optimization of algorithms.

Polystyrene nanoparticles enhance the adverse effects of di-(2-ethylhexyl) phthalate on male reproductive system in mice.

Coexposure of nanoplastics (NPs) with other pollutants adsorbed from the surroundings has received extensive attention. Currently, the combined effects of NPs and plasticizers remain unclear. Di-(2-ethylhexyl) phthalate (DEHP) is a commonly used plasticizer that has raised much concern owing to its ubiquitous pollution and endocrine-disrupting potential. This study aimed to investigate the toxic effects on the male reproductive system upon coexposure to NPs and DEHP. The C57BL/6J mice were orally administrated with polystyrene nanoparticles (PSNPs), DEHP or both for 35 days to evaluate their effects on sperm quality, histology of testes and epididymides, testicular transcriptomic characteristics as well as expression of some important genes in the epididymides. The low-dose PSNPs used here did not induce significant changes in sperm quality, while DEHP alone or cotreatment with DEHP and PSNPs caused notable impairment, mainly manifesting as decreased sperm quality and aberrant structure of the testis and epididymis. Moreover, enhanced toxic effects were found in the cotreatment group when compared with the individual DEHP treatment group, as manifested by more obvious alterations in the sperm parameters as well as histological changes in the testis and epididymis. Testicular transcriptomic analysis revealed differential regulation of genes involved in immune response, cytoplasmic pattern recognition receptor signaling pathways, protein ubiquitination, oxidative stress, necrotic cell death, ATP synthesis and the cellular respiratory chain. RT-qPCR verified that the expression patterns of Cenpb, Crisp1 and Mars were changed in testes, and genes relevant to epididymal function including Aqp9 and Octn2 were downregulated in epididymides, particularly in the cotreatment group. Collectively, our results emphasize that DEHP at an environmentally relevant dose can induce male reproductive toxicity, and PSNPs may aggravate the toxic effects.

Comprehensive Diagnostic Medical System Based on Notch1 Signaling Pathway to Inhibit the Growth of Small-Cell Lung Carcinoma.

With the gradual application of big data and other technologies to the medical field, more and more people tend to get online medical services. This article mainly studies the comprehensive diagnostic medical system based on Notch1 signaling pathway to inhibit the growth of small-cell lung carcinoma. In the experiment, we used the rapid thawing method to recover the cells and took the logarithmic growth phase cells for cell passage. We calculated the cell concentration and diluted the cells according to the experimental requirements. According to the standard curve, the corresponding sample protein concentration was calculated; at the same time, the Trizol method was used to extract the total RNA, the NanoDrop8000 spectrophotometer was used to determine the RNA concentration, and the RNA quality was detected by agarose gel electrophoresis. We used immunohistochemical staining to complete the staining of lung cancer cells. Finally, black box testing was used to test the functional modules of the system. Experimental data show that the accuracy rate of data obtained by the system reaches 98%, which greatly facilitates doctors and patients. The results show that the system has good ease of use and reliability and improves the diagnosis and treatment of hospital patients.

Analysis of m6A modification regulators in the substantia nigra and striatum of MPTP-induced Parkinson's disease mice.

Parkinson's disease (PD) is one of the most common neurodegenerative disorders of aging that impairs predominately dopaminergic neurons. N6-methyladenosine (m6A) is the most prevalent form of internal RNA modification in eukaryotes and it plays an essential role in normal brain development and neurodegenerative diseases. The m6A status is dynamically modulated by diverse types of genes called "writers", "erasers" and "readers". However, whether these m6A regulators are perturbed in PD remains poorly understood. To clarify this point, we established a PD mouse model using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The motor as well as learning and memory ability of mice were evaluated through and rotarod and Y maze spontaneous alternation tests. Morphological characteristics of tyrosine hydroxylase (TH)-positive cells were visualized using immunohistochemistry, while expressions of alpha-synuclein (α-syn) and TH were determined by using western blot. Furthermore, the expressions of the m6A regulators in the substantia nigra and striatum were evaluated by using qRT-PCR and western blot. As a result, the MPTP-induced PD mice suffered from learning and memory as well as motor defects. Additionally, there were significant TH+ neuron losses in the substantia nigra and striatum of MPTP-injected mice. In the PD mice, proteins including ALKBH5, IGF2BP2 were up-regulated in the substantia nigra, while YTHDF1 and FMR1 was down-regulated. For the striatum, FMR1 and CBLL1 were up-regulated, while IGF2BP3, METTL3 and RBM15 were down-regulated. The expression of genes at the mRNA level were partially in accordance with the protein changes. These findings indicate the m6A regulators may participate in PD pathogenesis.

Polystyrene nanoparticles aggravate the adverse effects of di-(2-ethylhexyl) phthalate on different segments of intestine in mice.

Emerging evidence indicates that nanoplastics (NPs) can transport organic pollutants such as di-(2-ethylhexyl) phthalate (DEHP) into organisms and induce adverse health effects. Nevertheless, the toxic effects of NPs combined with DEHP on mammalian intestine are still unclear. In this study, the C57BL6J mice were exposed to polystyrene nanoparticles (PSNPs), DEHP or them both for 30 days to determine their effects on different segments of intestine and the gut microbiota. As a result, DEHP alone or co-exposure to DEHP and PSNPs induced histological damages in all intestinal parts, mainly manifested as the decreased villus lengths, increased crypt depths in the duodenum, jejunum and ileum and decreased villus counts accompanied with decreased epithelial area in the colon. Moreover, decreased mucus coverage, down-regulated Muc2 expression levels as well as the broken tight junctions were observed in intestinal epithelium of mice, particularly obvious in the co-treatment groups. In general, as manifested by greater alterations in most of the parameters mentioned above, simultaneously exposed to PSNPs and DEHP seemed to induce enhanced toxic effects on intestine of mouse when compared with DEHP alone. Furthermore, the altered community composition of gut microbiota might at least partially contribute to these abnormalities. Overall, our results highlight the aggravated toxicity on different segments of intestine in mammalians due to co-exposure of PSNPs and DEHP, and these findings will provide valuable insights into the health risk of NPs and plastic additives.

Did the modern Yellow River form at the Mid-Pleistocene transition?

The incision of the Sanmen Gorge marks the birth of the modern Yellow River, but its timing varies from the late Miocene-early Pliocene to the late Pleistocene (∼0.15 Ma), and the suggested forcing mechanisms vary from the uplift of the Tibetan Plateau to global climate change. Here, we report sedimentologic, geochronologic, and provenance data from a drill core near the Sanmen Gorge, the last gorge along the main course of the Yellow River. Our results indicate that typical river channel deposits, with detritus from the Ordos Block in the upstream regions, started to accumulate in the Sanmen Gorge at ∼1.25 Ma. When integrated with river terrace evidence from the upstream and downstream regions, the results provide robust evidence that the final integration of the modern Yellow River occurred at ∼1.25 Ma, consistent with the beginning of the Mid-Pleistocene transition (MPT). We propose that the accelerated lowering of eustatic sea level during the MPT may play as important a role as tectonism in driving the birth and evolution of the modern Yellow River.

Repression of autophagy leads to acrosome biogenesis disruption caused by a sub-chronic oral administration of polystyrene nanoparticles.

As a new widespread contaminant, nanoplastics (NPs) pose a potential risk to human health. Nevertheless, the adverse effects of NPs on the male reproductive system are poorly understood. In this study, we aimed to determine the effects of polystyrene nanoplastics (PS-NPs) (50 nm) on sperm quality, with a focus on the acrosome defects. After 35 days of intragastric administration, sperm quality was decreased and testicular structures were impaired in mice exposed to PS-NPs in both the medium (1.0 mg/kg) and high dose (10 mg/kg) groups. No significant changes were observed in the low dose (0.2 mg/kg) group. Meanwhile, acrosome parameters including acrosome integrity and acrosome reaction were decreased after the administration of PS-NPs. These findings were consistent with the disruption of acrosome biogenesis, as identified by the changed testicular ultrastructure. Additionally, the findings were further validated using seven marker genes (Gba2, Pick1, Gopc, Hrb, Zpbp1, Spaca1 and Dpy19l2) essential for acrosome formation, which showed that two of these genes (Gopc and Dpy19l2) were significantly down-regulated. Moreover, repressed autophagy was observed in the testes of PS-NPs-exposed mice based on autophagy-related protein expression. This phenomenon was further verified in GC-2spd cells treated with PS-NPs (50 µg/mL, 100 µg/mL, 200 µg/mL for 24 h). The potential role of autophagy in such acrosome defects was explored by using the autophagy inhibitor 3-methyladenine (3-MA), autophagy activator rapamycin or beclin-1 siRNA. The results showed that Golgi-associated vesicle disorganization was exacerbated with the 3-MA and beclin-1 siRNA pretreatments, but decreased with the rapamycin pretreatment, and the expression of GOPC and DPY19L2 was also altered. These results indicated that autophagy might be involved in the PS-NPs-induced acrosome lesions based on the regulation of two key acrosome-formation proteins, GOPC and DPY19L2. Altogether, our results will provide new insights into the PS-NPs-induced male reproductive impairment.

Pathological analysis of hesperetin-derived small cell lung cancer by artificial intelligence technology under fiberoptic bronchoscopy.

Lung cancer is one of the most common tumors. There are 1.8 million new cases worldwide each year, accounting for about 13% of all new tumors. Lung cancer is the most important cause of cancer-related deaths. 1.4 million people die of lung cancer each year. This article uses artificial intelligence technology to analyze the pathology of hesperetin-derived small cell lung cancer under fiberoptic bronchoscopy. This article takes 48 lung slice samples as the research object. Among them, 36 cases of lung small cell carcinoma have history slices from Lhasa City Institute of Biology, the patient has complete cases, and the other 12 normal lung slices come from Xinjiang Biotechnology Laboratory. In this paper, the above-mentioned 36 lung cancer slices became the study group, and 12 normal slices became the reference group. This article presents a method for hesperetin-fiber bronchoscope to study the pathological mechanism of lung small cell carcinoma (H-FBS), which is used to study slices. The above-mentioned 48 samples were taken for slice observation. First, the 48 slices were technically tested by artificial intelligence fiber bronchoscope combined with hesperetin derivatives, and then the slice observation results were verified by CTC technology. In addition, in each step, the C5orf34 in the tissue is detected separately, which is beneficial to adjust the content of C5orf34 so that the treatment of lung cancer can control the development of lung cancer under fiberoptic bronchoscopy. Experimental results show that the diagnostic accuracy rate of this method is 97.9%, which is higher than that of lung biopsy (89%); compared with multiple CTC detection, the cost is low and the time is shor.

Downregulation of miR-29b-3p aggravates podocyte injury by targeting HDAC4 in LPS-induced acute kidney injury.

Sepsis is a severe organ dysfunction disease, usually accompanied by acute kidney injury (AKI). miR-29b-3p was inhibited in sepsis-induced AKI, while its role in AKI was unclear. Therefore, this study determined the role of miR-29b-3p in sepsis-induced AKI, and investigated its underlying mechanism. In this study, the AKI model was established through injecting with lipopolysaccharides (LPS) intraperitoneally. In LPS challenged mice, serum blood urea nitrogen and creatinine were increased, and renal tissues pathological damage was induced. Besides, miR-29b-3p was declined in LPS-induced AKI mice and podocytes. In addition, HDAC4 was elevated in LPS-treated podocytes. Furthermore, upregulated miR-29b-3p attenuated LPS-induced mice podocyte injury, and HDAC4 was identified as a direct target of miR-29b-3p. Moreover, overexpression of miR-29b-3p attenuated LPS-induced AKI in mice. In conclusion, miR-29b-3p was inhibited in LPS-induced AKI. Downregulation of miR-29b-3p aggravated podocyte injury through targeting HDAC4 in LPS-induced AKI. miR-29b-3p may act as a valuable target for AKI therapy.

Suppressing LncRNA HOXA-AS3 by CRISPR-dCas9 inhibits pancreatic cancer development.

The lncRNA HOXA-AS3 has been reported as a potential oncogene in tumors. Nevertheless, the molecular mechanism of HOXA-AS3 in pancreatic cancer (PC) progression remains unknown. We performed quantitative real-time (qRT) PCR assay to detect the expression levels of HOXA-AS3, miR-29c in PC specimens. Then, we transfected sgRNA-HOXA-AS3, miR-29c mimics, miR-29c inhibitors, or vector-CDK6 plasmids into PC cell lines to regulate the expression levels of HOXA-AS3, miR-29c or CDK6. Luciferase reporter assay was performed to identify the correlations among miR-29c, HOXA-AS3 and 3' UTR of CDK6.The ability of cell proliferation was assessed by cell counting and subcutaneous tumor growth assay. HOXA-AS3 level was upregulated in PC, and its knockdown suppressed PC cells proliferation, whereas miR-29c antagonized the regulatory effect of HOXA-AS3 knockdown by directly binding to HOXA-AS3.Moreover, CDK6 was a target of miR-29c and miR-29c exerted anti-proliferation effects through inhibiting CDK6. HOXA-AS3 could accelerate the growth of PC cells partially by regulating the miR-29c/CDK6 axis, which could be used as a potential therapeutic target in CRISPR-mediated PC treatment.

Metformin inhibits epithelial-mesenchymal transition of oral squamous cell carcinoma via the mTOR/HIF-1α/PKM2/STAT3 pathway.

Epithelial-mesenchymal transition (EMT) serves an important role in the formation and development of various types of cancer, including oral squamous cell carcinoma (OSCC). Metformin, used for treating type 2 diabetes, has been revealed to exert an anticancer effect in various types of cancer, including liver, breast and colorectal cancer. However, its role in the EMT of OSCC has been rarely reported. Therefore, the present study aimed to investigate the effects of metformin on EMT and to identify its underlying mechanism in OSCC. Firstly, EMT was induced in CAL-27 cells using CoCl2. Subsequently, the effects of metformin on cell viability, migration and xenograft growth were evaluated in vitro and in vivo. Reverse transcription-quantitative PCR was performed to detect the expression levels of E-cadherin, vimentin, snail family transcriptional repressor 1, mTOR, hypoxia inducible factor 1α, pyruvate kinase M2 and STAT3. The results demonstrated that metformin abolished CoCl2-induced cell proliferation, migration, invasion and EMT. Moreover, metformin reversed EMT in OSCC by inhibiting the mTOR-associated HIF-1α/PKM2/STAT3 signaling pathway. Overall, the present findings characterized a novel mechanism via which metformin modulated EMT in OSCC.

LncRNA NEAT1 promotes apoptosis and inflammation in LPS-induced sepsis models by targeting miR-590-3p.

Sepsis is a complication of infection caused by disease or trauma. Increasing evidence have shown that long noncoding RNAs (lncRNAs) are involved in the regulation of sepsis. However, the mechanism of lncRNA nuclear enriched abundant transcript 1 (NEAT1) in the regulation of sepsis progression remains to be elucidated. Lipopolysaccharide (LPS) was used to induce a sepsis cell model. The expression levels of NEAT1 and microRNA (miR)-590-3p were determined by reverse transcription-quantitative PCR. Cell viability and apoptosis were detected using Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. Western blot analysis was performed to evaluate the levels of apoptosis- and NF-κB signaling pathway-related proteins. The concentration of inflammatory cytokines was determined using ELISA. In addition, dual-luciferase reporter assay, RNA immunoprecipitation and biotin-labeled RNA pull-down assay were performed to verify the interaction between NEAT1 and miR-590-3p. The results showed that NEAT1 was highly expressed in patients with sepsis and LPS-induced H9c2 cells. Knockdown of NEAT1 decreased LPS-induced cell apoptosis and inflammation response in H9c2 cells. Meanwhile, miR-590-3p showed decreased expression in sepsis, and its overexpression could relieve LPS-induced H9c2 cell damage. Further experiments revealed that NEAT1 could sponge miR-590-3p. Knockdown of miR-590-3p reversed the inhibitory effect of NEAT1 knockdown on LPS-induced H9c2 cell damage. Additionally, the NEAT1/miR-590-3p axis could regulate the activity of the NF-κB signaling pathway. To conclude, lncRNA NEAT1 accelerated apoptosis and inflammation in LPS-stimulated H9c2 cells via sponging miR-590-3p. These findings may provide a new strategy for the treatment of sepsis.

High dose erythropoietin promotes functional recovery of rats following facial nerve crush.

Erythropoietin (Epo) has neuroprotective activity in a variety of settings. Thus, we investigated whether Epo has a role in the functional recovery of rats after facial nerve injury. The right facial nerve of 24 Wistar rats (6 wks old) was crushed twice at the level of the stylomastoid foramen, for 30 s each time, using jeweler's forceps held perpendicular to the nerve. The left facial nerve did not undergo the surgical lesion. The rats were randomly divided into 4 groups: (group 1) the control group (placebo, treated with saline); and groups treated with Epo at a dose of 1,000 U/kg body weight (group 2), 5,000 U/kg body weight (group 3), and 10,000 U/kg body weight (group 4). The Epo and saline were administered subcutaneously pre-operatively and treatment was repeated every 24 h for the first 2 weeks after the operation. Behavioral recovery from facial paralysis was measured daily, beginning 1 day after surgery, until full recovery of the eye blink reflex and whisker movements were observed. The average recovery times for the full blink reflex and whisker movements were significantly shorter (about 2-3 days) in rats treated with a high dose Epo (5,000, 10,000 U/kg body weight) compared to the placebo-treated rats (p<0.05). There was no significant difference between low dose Epo-treated rats (1,000 U/kg body weight) and the placebo-treated rats. These results suggest that high dose Epo can promote the functional recovery of rats following facial nerve injury. Further studies are warranted to probe alternative treatment schedules (dose, mode of administration), underlying histological mechanisms and combination treatment with additional neuroprotective factors.

Recognizing the Effects of Language Mode on the Cognitive Advantages of Bilingualism.

For bilinguals, it is argued that a cognitive advantage can be linked to the constant management and need for conflict resolution that occurs when the two languages are co-activated (Bialystok, 2015). Language mode (Grosjean, 1998, 2001) is a significant variable that defines and shapes the language experiences of bilinguals and consequently, the cognitive advantages of bilingualism. Previous work, however, has not sufficiently tested the effects of language mode on the bilingual experience. In this brief conceptual analysis, we discuss the significance of language mode in bilingual work on speech perception, production, and reading. We offer possible explanations for conflicting findings and ways in which future work should control for its modulating effects.

Establishment of OPG Transgenic Mice and the Effect of OPG on Bone Microarchitecture.

Osteoprotegerin (OPG) plays a determinant role in regulating bone metabolism, but the effect of OPG on bone microarchitecture needs to be further elucidated. We attempted to construct pCI-hOPGp-mOPG vector containing human OPG promoter and FLAG tag and to microinject vector into fertilized zygotes from C57BL/6J × CBA mice to prepare transgenic mice. The OPG transgenic positive mice were identified by PCR and western blotting. Twelve-week-old OPG transgenic mice (OPG-Tg mice) and wild-type mice (WT mice) were utilized in the study of bone microarchitecture. Microcomputed tomography (micro-CT) data showed that compared with WT mice, the tibia of OPG-Tg mice showed an increased volumetric BMD (vBMD), tissue BMD (tBMD), trabecular thickness (Tb.Th), and trabecular number (Tb.N), and a decreased trabecular separation (Th.Sp) (P < 0.05) . The cortical bone microarchitecture parameters, such as cortical area (Ct.Ar), cortical thickness (Ct.Th), cortical BMD (Ct.BMD), cortical BMC (Ct.BMC), BMD, and BMC of femur, were increased, and the inner perimeter (In.Pm) was decreased, in OPG-Tg mice, compared to those in WT mice (P < 0.05). The established OPG transgenic mouse model could be valuable for further studying the biological significance and gene regulation of OPG in vivo.

[The expression of collagen I during mandibular fracture healing with the inferior alveolar nerve mutilated].

OBJECTIVE: To establish a stable mandibular fracture model with inferior alveolar nerve (IAN) mutilated for exploring the IAN influence to the expression of collagen I during mandibular fracture. METHODS: Japanese white rabbits were selected and IAN was liberated. The right was leaven as it is and the left was cut off, then 2 mm x 5 mm fracture model was made, and HE staining and chromotropic acid 2R-bright green staining and in situ hybridization of collagen I mRNA were made to detect mandibular fracture healing and collagen I mRNA expression. RESULTS: Fracture healing was smooth in IAN conserved side, but delayed in the IAN mutilated side. Collagen I in situ hybridization showed that at one week and two weeks after surgery there were obvious differences between the two sides (P < 0.05), but no difference at three weeks after surgery and four weeks (P > 0.05). CONCLUSION: The IAN could regulate mandibular fracture healing, and influence collagen I mRNA earlier expression.