CircHERC1 promotes non-small cell lung cancer cell progression by sequestering FOXO1 in the cytoplasm and regulating the miR-142-3p-HMGB1 axis.

BACKGROUND: Noncoding RNAs such as circular RNAs (circRNAs) are abundant in the human body and influence the occurrence and development of various diseases. Non-small cell lung cancer (NSCLC) is one of the most common malignant cancers. Information on the functions and mechanism of circRNAs in lung cancer is limited; thus, the topic needs more exploration. The purpose of this study was to identify aberrantly expressed circRNAs in lung cancer, unravel their roles in NSCLC progression, and provide new targets for lung cancer diagnosis and therapy. METHODS: High-throughput sequencing was used to analyze differential circRNA expression in patients with lung cancer. qRT‒PCR was used to determine the level of circHERC1 in lung cancer tissues and plasma samples. Gain- and loss-of-function experiments were implemented to observe the impacts of circHERC1 on the growth, invasion, and metastasis of lung cancer cells in vitro and in vivo. Mechanistically, dual luciferase reporter assays, fluorescence in situ hybridization (FISH), RNA immunoprecipitation (RIP) and RNA pull-down experiments were performed to confirm the underlying mechanisms of circHERC1. Nucleocytoplasmic localization of FOXO1 was determined by nucleocytoplasmic isolation and immunofluorescence. The interaction of circHERC1 with FOXO1 was verified by RNA pull-down, RNA immunoprecipitation (RIP) and western blot assays. The proliferation and migration of circHERC1 in vivo were verified by subcutaneous and tail vein injection in nude mice. RESULTS: CircHERC1 was significantly upregulated in lung cancer tissues and cells, ectopic expression of circHERC1 strikingly facilitated the proliferation, invasion and metastasis, and inhibited the apoptosis of lung cancer cells in vitro and in vivo. However, knockdown of circHERC1 exerted the opposite effects. CircHERC1 was mainly distributed in the cytoplasm. Further mechanistic research indicated that circHERC1 acted as a competing endogenous RNA of miR-142-3p to relieve the repressive effect of miR-142-3p on its target HMGB1, activating the MAPK/ERK and NF-κB pathways and promoting cell migration and invasion. More importantly, we found that circHERC1 could bind FOXO1 and sequester it in the cytoplasm, adjusting the feedback AKT pathway. The accumulation of FOXO1 in the cytosol and nuclear exclusion promoted cell proliferation and inhibited apoptosis. CircHERC1 is a new circRNA that promotes tumor function in NSCLC and may serve as a potential prognostic biomarker and therapeutic target for NSCLC. CONCLUSIONS: CircHERC1 is a new circRNA that promotes tumor function in NSCLC and may serve as a potential diagnosis biomarker and therapeutic target for NSCLC. Our findings indicate that circHERC1 facilitates the invasion and metastasis of NSCLC cells by regulating the miR-142-3p/HMGB1 axis and activating the MAPK/ERK and NF-κB pathways. In addition, circHERC1 can promote cell proliferation and inhibit apoptosis by sequestering FOXO1 in the cytoplasm to regulate AKT activity and BIM transcription.

circTAB2 inhibits lung cancer proliferation, migration and invasion by sponging miR-3142 to upregulate GLIS2.

Circular RNAs (circRNAs) are a specialized circular structure, are deregulated in cancers and play essential roles in biological processes involved in tumor progression. However, the mechanism by which circRNAs affect lung tumorigenesis and progression remains largely unexplored. To investigate the role of circRNA in lung cancer, circRNA expression profile was screened by bioinformatics analysis. The levels of circTAB2, miR-3142, and GLIS family zinc finger 2 (GLIS2) were measured by quantitate real-time (qRT-PCR) or western blot. Cell proliferation, apoptosis, migration and invasion were detected by EdU, flow cytometry, and transwell assays, respectively. Bioinformatics, western blot, RIP, pull down, dual luciferase reporter and rescue experiments were used to verify the direct relationship between miR-3142 and circTAB2 or GLIS2. The xenograft assays were used to assess the role of circTAB2 in vivo.CircTAB2 exhibited low expression in cancer tissues. Gain and loss-of-function assays indicated that circTAB2 could inhibit cell proliferation, migration and invasion. Functional studies revealed that circTAB2 acted as a miRNA sponge, directly interacted with miR-3142 and consequently regulated GLIS2 /AKT. Taken together, circTAB2 serves as an inhibitory role in lung cancer through a novel circTAB2 /miR-3142 /GLIS2 /AKT pathway and could be exploited a novel marker in lung cancer.

[Screening of an Armillaria gallica strain for Gastrodia elata cultivation].

This study aims to screen a strain from Armillaria for the cultivation of Gastrodia elata. Specifically, Armillaria strains were isolated from different producing areas of G. elata and identified. Based on the growth characteristics of the strains and the experiment on the cultivation of G. elata, an optimal A. gallica strain was screened out. The specific process is as follows. The fungus-gro-wing materials of G. elata were collected from four producing areas and the Armillaria strains were isolated(G,Y,S,H). The strains were then identified based on morphological observation and phylogeny analysis and the commonly used strains were determined. The sucrase genotypes of the strains were identified according to our previous research findings, and the growth characteristics of the strains, such as growth rate, diameter, dry weight, and polysaccharide content of the rhizomorphs, were measured. According to the biological characteristics and sucrase genotypes, two strains were selected for the cultivation of G. elata. The tuber yield and the content of gastrodin and p-hydroxybenzyl alcohol in the tuber of G. elata were measured to select the optimal strain. The results showed that the four strains were all A. gallica. The rhizomorphs of strains G and H of the same sucrase genotype had larger/higher length, growth rate, diameter, branch number, dry weight, and polysaccharide content than those of strains S and Y of the same sucrase genotype. The tuber yield and the total content of gastrodin and p-hydroxybenzyl alcohol in tuber of G. elata cultivated with strain H were 6.528 kg·m~(-2) and 0.566%, respectively, which were 4.58 and 1.30 folds those of G. elata cultivated with strain S. Strains H and S were screened out from four strains of A. gallica based on the growth characteristics and sucrase genotype. According to the tuber yield and content of total gastrodin and p-hydroxybenzyl alcohol in the tuber of G. elata, strain H was identified as the optimal one. The findings in this study are expected to lay a basis for cultivating G. elata with high yield and quality of tubers.

Identification of the stable and reactive metabolites of tetrahydropiperine using ultrahigh-performance liquid chromatography combined with diode-array detection and high-resolution mass spectrometry.

RATIONALE: Tetrahydropiperine is one of the natural arylpentanamide compounds isolated from Piper nigrum L., which has been demonstrated to have insecticidal activity. The aim of this study was to investigate the metabolic profiles of tetrahydropiperine in mouse, rat, dog, monkey and human hepatocytes. METHODS: The in vitro metabolism of tetrahydropiperine was elucidated via incubation with hepatocytes for 2 h at 37°C. The samples were analyzed using ultrahigh-performance liquid chromatography combined with diode-array detection and high-resolution tandem mass spectrometry operated in positive electrospray ionization mode. The structures of the metabolites were characterized using their retention times and their tandem mass spectrometric product ions. RESULTS: A total of 20 metabolites were detected and their structures were proposed. These metabolites were formed mainly through the following pathways: (1) 1,3-benzodioxole ring opening to form a catechol derivative (M12), which was prone to glucuronidation (M6 and M8), methylation (M17) and glutathione (GSH)-derived conjugation through an ortho-quinone intermediate (M4) or via an aldehyde intermediate (M7); (2) dehydrogenation to form a piperanine (M15), which was subsequently subject to hydroxylation (M2 and M5) and GSH conjugation (M10 and M11) via Michael addition; (3) hydroxylation (M13, M14, M16, M18 and M19); and (4) direct GSH conjugation through an aldehyde intermediate (M3). CONCLUSIONS: The major metabolic pathways of tetrahydropiperine were hydroxylation, dehydrogenation, methylation, GSH conjugation and glucuronidation. Tetrahydropiperine was bioactivated through ortho-quinone, Michael receptor and aldehyde intermediates.

[Identification of Gastrodia elata and its hybrid by polymerase chain reaction].

Gastrodia elata is a kind of traditional Chinese medicinal materials and has good medicinal value. G. elata is divided into five varieties, which includes G. elata f. elata(proto variant), G. elata f. glauca, G. elata f. viridis, G. elata f. flavid and G. elata f. alba. Among them, G. elata f. elata and G. elata f. glauca have excellent characteristics and higher contents of gastrodin and polysaccharides. The hybrid of G. elata f. elata and G. elata f. glauca is present in markets, but the characteristics between hybrid and parent are not obvious and distinguished quickly and accurately. The aim of this study is to establish a PCR specific PCR identification method, which can identify G. elata f. elata, G. elata f. glauca and their hybrid. Based on the re-sequencing results of G. elata, we screened for the single nucleotide polymorphism(SNP) variation sites, and designed two pairs of specific primers(W291-F/W291-R and H255-F/H255-R). We further collected G. elata f. elata, G. elata f. glauca and their hybrid samples from different regions, established and optimized PCR method, and investigated and verified their tolerance and applicability. The results showed that when the annealing temperature was 48 ℃ and the number of cycles was 33, 255 bp specific band were obtained from G. elata f. glauca and hybrid by using specific primers W291-F/W291-R. When the annealing temperature was 51 ℃ and the number of cycles was 33, 291 bp specific band were obtained from G. elata f. elata and hybrid by using specific primers H255-F/H255-R. Our method could be used as a promising method to identify G. elata f. elata, G. elata f. glauca and their hybrid.

Electrocardiographic findings of acute total occlusion associated with a sub-occlusion involving the left anterior descending and the right coronary artery.

BACKGROUND: The ECG characteristics of simultaneous acute occlusion/sub-occlusion of two coronary arteries involving the left anterior descending (LAD) and right (RCA) coronary artery have been rarely described in the literature. METHODS: We present two patient cases, where one of the arteries was totally occluded and the other one had a sub-occlusion with severely limited flow to demonstrate the ECG characteristics of this severe presentation of acute coronary syndrome. RESULTS: Two ECG patterns suggested simultaneous occlusions of the RCA and LAD. One pattern was ST-segment elevation (STE) in lead III higher than in lead II with concomitant STE in leads V3-V4. The other pattern was STE in lead III higher than in lead II with the concomitant Dressler - de Winter ECG pattern in leads V2-V4. CONCLUSIONS: We present two ECG presentations of simultaneous RCA and LAD occlusion/sub-occlusion. We consider these ECG features as high-risk markers in acute ST-elevation myocardial infarction.

Predicting the outcome of acute pulmonary embolism by dynamic changes of the QRS complex in lead V1.

Electrocardiography can provide useful prognostic information in acute pulmonary embolism (APE). Several abnormal QRS changes in lead V1, including notched or fragmented QRS, incomplete or complete right bundle branch block (IRBBB or CRBBB) and the QR sign, which are associated with APE, are of prognostic significance. To illustrate this, we describe lead V1 QRS changes in combination with the clinical state of six APE patients. The dynamic ECG changes suggest that a change from a diminution of the S wave amplitude to notched S wave, next to RBBB and then to the QR sign indicate worsening of the patients' condition, and vice versa. Also, a diminution of the S wave amplitude in lead V1 associated with a final R' wave in the right precordial accessory leads indicates the possibility of hidden RBBB. Understanding dynamic QRS changes in APE may aid in risk stratification.

Design and synthesis of diverse Cd2+/Zn2+/Cu2+ coordination polymers tuned by dicarboxylate and auxiliary 1,4-bis(pyridin-4-ylmethyl)piperazine ligands with luminescence and Hirshfeld surface analyses.

Four new coordination polymers, including 1D, 2D and 3D structures, were synthesized via a hydrothermal method using Cd2+/Zn2+/Cu2+ metal salts as nodes. These polymers were formed through self-assembly of four different dicarboxylic acid ligands, namely adamantane-1,3-dicarboxylic acid (H2adc), glutaric acid (H2glu), 5-hydroxyisophthalic acid (H2hip) and fumaric acid (H2fum), in conjunction with the auxiliary ligand [1,4-bis(pyridin-4-ylmethyl)piperazine (bpmp). The corresponding formulae are [Cd3(adc)2(bpmp)Cl2(H2O)2]n (1), {[Cd2(glu)2(bpmp)2(H2O)2]·8H2O·2CH3OH}n (2), [Zn(hip)(bpmp)(H2O)]n (3) and [Cu(fum)(bpmp)(H2O)2]n (4). Single-crystal X-ray diffraction studies revealed that the Cd2+ centers in complex 1 all adopt a six-coordinate mode but two distinct {CdO2N2Cl2} and {CuO5Cl} units. The 3D network of complex 1 can be simplified to a binodal (4.6)-connected underlying net with the point symbol (3·42·5·62)4(32·62·72·88·10). Each Cd2+ cation in complex 2 adopts a seven-coordinate {CdO5N2} center, forming an asymmetric pentagonal bipyramidal coordination. Its stacking structure is formed by the interaction of hydrogen bonds between 2D supramolecular layers, with the adjacent layers exhibiting mirror symmetry. Each Zn2+ ion in complex 3 displays a {ZnO3N} four-coordinate unit. Its stacking structure is formed by one-dimensional [Zn(hip)(bpmp)(H2O)]n chains connected through hydrogen bonds. On the other hand, complex 4 features a Jahn-Teller distorted {CuO4N2} octahedral coordination. Subsequently, the thermal stability of these complexes was investigated. The solid-state fluorescence spectroscopy was employed to analyze complexes 1, 2 and 3. Additionally, a Hirshfeld surface analysis was performed on complex 3.

Synthesis, characterization, and methylene blue adsorption of multiple-responsive hydrogels loaded with Huangshui polysaccharides, polyvinyl alcohol, and sodium carboxyl methyl cellulose.

Huangshui (HS), a typical by-product of traditional Chinese Baijiu, has attracted more and more attention since its rich resources of polysaccharides. However, there's little information on hydrogels preparation by Huangshui polysaccharides (HSPs). A series of novel HSP-loaded hydrogels were synthesized using crude HSP (cHSP), polyvinyl alcohol (PVA), sodium carboxyl methyl cellulose (CMC), and in situ incorporation of Fe3O4 for methylene blue (MB) adsorption for the first time. Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), vibrating sample magnetometer (VSM) and texture tests indicated the paramagnetic hydrogels with porous structure were formed via physical crosslinking, and cHSP had no effect on hydrogel texture. Notably, cHSP enhanced the swelling capacity and MB adsorption ability of the hydrogels, and the corresponding maximum value was 38.67 g/g and 71.07 mg/g, respectively, when the additive amount of cHSP to the hydrogels was 2 % at 25°Ð¡. In addition, the swelling kinetics of hydrogels followed the Schott's second-order kinetics model, while MB adsorption fitted well with the Freundlich isotherm and pseudo-second-order model. Furthermore, intraparticle and film diffusion-controlled MB adsorption process. Significantly, cHSP amount could counteract the negative influence of high temperature on MB adsorption, and the prepared hydrogels could be reusable, demonstrating cHSP effectively promoted the properties of hydrogels and had potential application in MB removal. The adsorption mechanism of MB onto the hydrogels involved ion-exchange, hydrogen bonding, electrostatic interaction, and chemical reaction according to the above results together with the analysis by X-ray photoelectron spectroscopy (XPS) and energy-dispersive X-ray (EDX) characterizations, thermodynamic, etc.

Iron-doped novel Co-based metal-organic frameworks for preparation of bifunctional catalysts with an amorphous structure for OER/HER in alkaline solution.

A novel two-dimensional Co-MOF material {[Co(dptz)2(oba)2]·(DMF)2}n is prepared using mixed organic ligands, which exhibits both OER (oxygen evolution reaction) and HER (hydrogen evolution reaction) catalytic performance. The integration of an Fe dopant and amorphous interface into Co-MOF to improving the electrocatalytic performance of pristine MOFs (metal-organic frameworks) is demonstrated and the origin of the remarkable electrocatalytic performance of the catalyst is elucidated. The comprehensive characterization data of Fe@Co-MOFs illustrate that there is a crystallinity transition during the doping of Co-MOF, which increases the electron transfer rate of the material and ensures increased exposure of the ligand unsaturated active site on the surface, and modulates the electronic structure of the Co center in a synergistic manner. As a result, the optimized catalytic Fe@Co-MOF-3 with an amorphous structure exhibits outstanding electrocatalytic performance for the OER, with only 248 mV at a current density of 50 mA cm-2 and excellent stability after 11 h of testing in alkaline solution. Not only that, the HER was achieved with a low overpotential of 150 mV at 10 mA cm-2. The present work indicates that the as-synthesized Co-MOF and Fe@Co-MOFs offer prospects in developing electrocatalysts for water splitting.

Structure elucidation and intestinal barrier protection of an α-D-glucan in Huangshui.

WLY-0, as an α-D-glucan with a molecular weight (Mw) of 11.12 kDa, was successfully isolated and purified from Huangshui (HS). The results of methylation and NMR indicated that the mainchain of WLY-0 was (1 â†’ 4)-α-D-glucan, with side chains linking at O-6. Meanwhile, the surface morphology characterization showed that WLY-0 had an irregular flake-like morphology with a rough and uneven surface and varies in sizes from nanometers to microns. Furthermore, WLY-0 relieved the increased paracellular permeability of FD4 and decreased TEER challenged by LPS, meanwhile inhibited the production of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1ß) and up-regulated the expression of TJ protein (Occludin, Claudin-1, ZO-1, and JAM-A) in Caco-2 cells, so to improve the intestinal barrier function. Our findings about the structural characteristics and biological activities of WLY-0 provided a scientific foundation for the utilization of HS as a potent source of an effective adjuvant in intestinal barrier injury treatment.

Hsa_circ_0004674 promotes osteosarcoma doxorubicin resistance by regulating the miR-342-3p/FBN1 axis.

BACKGROUND: The occurrence of chemoresistance is a common problem in tumor treatment. Circular RNA (circRNA) has been confirmed to be related to tumor chemoresistance. However, the role and the underlying molecular mechanism of hsa_circ_0004674 in the chemoresistance of osteosarcoma (OS) are still unclear. METHODS: The expression of hsa_circ_0004674, miR-342-3p, and fibrillin-1 (FBN1) was determined by qRT-PCR. Cell counting kit 8 assay was used to evaluate the doxorubicin (DXR) resistance of cells. The proliferation and apoptosis of cells were measured using colony formation assay and flow cytometry. Western blot analysis was utilized to examine the protein levels of resistance markers, Wnt/ß-catenin pathway markers and FBN1. The interaction between miR-342-3p and hsa_circ_0004674 or FBN1 was confirmed by dual-luciferase reporter assay and RNA pull-down assay. Moreover, animal experiments were performed to assess the effect of hsa_circ_0004674 silencing on the DXR sensitive of OS in vivo. RESULTS: The upregulated hsa_circ_0004674 was found in DXR-resistant OS tissues and cells. Knockdown of hsa_circ_0004674 could inhibit the DXR resistance of OS cells in vitro and promote the DXR sensitive of OS tumors in vivo. In addition, we discovered that hsa_circ_0004674 could sponge miR-342-3p, and miR-342-3p could target FBN1. MiR-342-3p inhibitor could reverse the inhibition effect of hsa_circ_0004674 knockdown on the DXR resistance of OS cells. Similarly, the suppressive effect of miR-342-3p on the DXR resistance of OS cells also could be reversed by FBN1 overexpression. Furthermore, we revealed that hsa_circ_0004674 silencing inhibited the activity of Wnt/ß-catenin pathway by the miR-342-3p/FBN1 axis. CONCLUSION: Hsa_circ_0004674 facilitated the DXR resistance of OS through Wnt/ß-catenin pathway via regulating the miR-342-3p/FBN1 axis, suggesting that hsa_circ_0004674 was a promising target for the chemoresistance of OS.

Evaluation of serum cysteine-rich protein 61 levels in patients with coronary artery disease.

AIM: The aim is to evaluate serum cysteine-rich protein 61 (Cyr61) levels in patients with coronary artery disease (CAD). PATIENTS & METHODS: Serum Cyr61 levels were measured in 180 patients with CAD and 74 participants without CAD. RESULTS: Serum Cyr61 levels were significantly higher in CAD patients. Patients with acute coronary syndrome showed significantly higher Cyr61 than those with stable angina pectoris. Serum Cyr61 levels in complex lesion group were significantly higher. Serum Cyr61 was positively correlated with Gensini score and C-reactive protein. Multivariable logistic regression analyses demonstrated that serum Cyr61 levels were independently correlated with the existence of CAD (p = 0.01). CONCLUSION: Our study suggested Cyr61 as a potential biomarker in characterizing CAD and therapeutic target for CAD.

[Observation on effects of hepatitis B vaccine immunization for 12 years in children in Beijing].

OBJECTIVE: To evaluate the long-term protective effects of hepatitis B vaccine after immunizing to the children for 12 years in Beijing. METHODS: The multiple stratified cluster sampling was used in this epidemiological survey. The sampling children's blood serum HBsAg, anti-HBs and anti-HBc were checked and measured by the solid phase radioimmunoassay (SPRIA). The serological level of these index and the causes of the children with HBsAg positive were analyzed. RESULTS: There were 2,419 cases 3-12 years-old children immunized with the hepatitis B vaccine in infant period were surveyed and the total HBsAg positive rate was 0.52%. The vaccine protective rate was 88.45% (95% CI: 65.67%-97.89%). The total anti-HBc positive rate was 2.21%, being no statistical significance among the age groups. The average anti-HBs positive rate of 3-6 years-old children immunized with gene recombining vaccine was 38.79% and descending greatly following the age's dropping. The geometric means of anti-HBs serological titer (GMT) was 52.83 mIU/ml, showing no statistical significance among the age groups. The average anti-HBs positive rate of 6-12 years-old children immunized with the blood rooting vaccine was 50.79%. The geometric means of anti-HBs serological titer (GMT) was 61.51 mIU/ml. There were no statistical significances among the age groups. Among the HBsAg positive children, more than 50% of the children's mothers were HBsAg positive also. CONCLUSIONS: The protective effects given by immunization were significant after the hepatitis B vaccine vaccination for 12 years in Beijing. The booster immunization was not necessary, because the HBsAg positive rate didn't ascend obviously as the immunization time prolonging. As the anti-HBs positive rate of children who were immunized by the gene recombining vaccine might be descending following the age's dropping greatly, we should strengthen the serological surveillance of hepatitis B. The main cause that the children became the HBsAg carrier should be a vertical transmission.

[Epidemiological effect of hepatitis B immunization among newborn babies in Beijing].

OBJECTIVE: To evaluate the epidemiological effect of hepatitis B immunization among newborn babies in Beijing. METHODS: A multistage sampling method was used for the collection of immunization cards, field epidemiological survey on hepatitis B virus (HBV) immunization of children, analysis of infectious disease reports. HBsAg, anti-HBs and anti-HBc levels were detected by solid phase radioimmunoassays (SPRIA). RESULTS: The incidence of hepatitis B in children of 0 to 14 years at the beginning of HBV immunization was 18.59 to 20.52/100,000, and declined to 0.39 to 2.38/100,000 in 2000 (chi2 = 58.26, P < 0.01). The HBsAg carrying rate of the children decreased from 2.82% to 0.60%, about 80.00% after vaccination (chi2 = 10.75, P < 0.01). CONCLUSION: Hepatitis B vaccination of newborn babies is an effective measure for prevention and control of hepatitis B virus infection.

[Study on the afficacy of hepatitis B immunization among youngsters in Beijing].

OBJECTIVE: To evaluating the epidemiological effect after hepatitis B immunization among youngsters in Beijing. METHODS: A multistage sampling method was used for data collection from immunization cards, reports on field epidemiological survey on hepatitis B virus (HBV) immunization of youngsters and the analysis of infectious diseases. HBsAg, anti-HBs and anti-HBc levels were detected by solid phase radioimmunoassays (SPRIA). RESULTS: The average incidence rates of hepatitis B in 10 - 19 years-olds prior to HBV immunization was 12.11-16.89/100 000, while declined to 1.78-10.95/100 000 (chi(2) = 10.71 - 60.45, P < 0.01). HBsAg carrying rate of the youngsters decreased from 6.34% to 1.37% (80.00%) after vaccination (chi(2) = 14.16, P < 0.01). CONCLUSION: Hepatitis B vaccination among youngsters is an effective measure for the prevention and control of hepatitis B virus infection.