Objective To investigate the contamination of four polycyclic aromatic hydrocarbons (PAHs) in domestic pre-packaged edible vegetable oil sold in Henan Province. Methods A total of 212 domestic edible vegetable oil samples from Henan Province were collected. The contents of four polycyclic aromatic hydrocarbons (benzo[a]anthracene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene, defined as PAH4) were determined by HPLC and gas chromatography-mass spectrometry in 2019 and 2020, respectively. Results The content of PAH4 in domestic pre-packaged edible vegetable oils ranged from 0.34 μg/kg to 26.6 μg/kg, the detection rate was 83.49%, and the over-standard rate was 3.30% according to EU standard. In terms of food category, the peanut oil samples were most seriously contaminated with PAH4. From the point of view of product grade and production process, with the decrease of sample quality index, the risk of PAH4 contamination in sesame oil increased. According to GB 2762-2017, the content of benzo (a) pyrene in all samples was qualified, while the peanut oil had the highest rate exceeding the EU standard, with the over-standard rate of 38.46%. Peanut oil had a higher risk of being contaminated by benzo [a] pyrene. Conclusion Peanut oil in domestic pre-packaged edible vegetable oil sold in Henan Province has a high risk of PAH4 contamination. With the decrease of sesame oil quality index, the risk of PAH4 contamination increases.

Objective To evaluate the clinical effect of endovascular stent placement to the treatment of intracranial internal carotid artery dissection.Methods Two patients with intracranial internal carotid artery dissection received the treatment of stent placement,and 1 patient with a dissection of the supra clinoid internal carotid artery received conventional anticoagulation treatment.Results Two patients with intracranial internal carotid artery dissection were given treatment of Apollo stent placement,of which 1 patient had improvement of left limb paresis,the score of NIHSS from 3 before operation to 2 after operation; the other one with episodic left limb weakness was not seen any attack after stent placement.Another one patient without stent placement receiving conventional anticoagulation treatment had some improvement of right limb paralysis.Conclusion The treatment of endovascular stent placement to the intracranial internal carotid artery dissection has better clinical efficacy and especially used for those patients with no effect to the conventional anticoagulation treatment.

Objective:To investigate the effect of allicin （ALL） on learning and memory ability of rats with vascular dementia （VD） and the possible mechanism. Method:The VD rats induced by modified bilateral common carotid artery occlusion （BCCAO） were randomly divided into the VD group， low- and high-dose ALL （ALL-L and ALL-H） groups， and the sham operation （S） group， with 15 rats in each group. In the ALL-L and ALL-H groups， ALL was injected into the femoral vein at 5 mg·kg^-1 and 20 mg·kg^-1， respectively， while the same volume of normal saline was injected in the S and VD groups， once a day， for two successive weeks. Morris water maze （MWM） was used to test the learning and memory ability of rats. Hematoxylin and eosin （HE） staining was conducted to observe the pathological changes in hippocampal tissue， followed by the detection of inflammatory factors tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and IL-1β as well as oxidative stress indexes malondialdehyde （MDA）， superoxide dismutase （SOD）， and glutathione peroxidase （GSH-Px） in rat hippocampus. The apoptosis of hippocampal cells was detected by TdT-mediated dUTP Nick end Labeling（TUNEL） assay. The expression levels of apoptosis and autophagy-related proteins cysteinyl aspartate-specific protease-3 （Caspase-3）， B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， microtubule-associated protein light chain 3Ⅱ （LC3Ⅱ）， LC3Ⅰ， and the mammalian homolog of yeast ATG6 （Beclin 1） in hippocampus were determined by Western blot. Result:The comparison with the VD group revealed that the learning and memory abilities of rats in the ALL-H and ALL-L groups were significantly improved （P<0.05）. The TNF-α， IL-6， IL-1β， and MDA levels in hippocampus were lowered （P<0.05）， whereas the SOD and GSH-Px activities were enhanced （P<0.05）. The apoptosis rates were declined （P<0.05）， with an even lower rate noticed in the ALL-H group （P<0.05）. The expression levels of Caspase-3， Bax， LC3Ⅱ/LC3Ⅰ ratio， and Beclin-1 in the ALL-H and ALL-L groups were significantly down-regulated in contrast to those in the VD group （P<0.05）， while that of Bcl-2 was up-regulated （P<0.05）. The ALL-H group exhibited better performances than the ALL-L group （P<0.05）. Conclusion:ALL could improve the learning and memory ability of VD rats to some extent， which may be attributed to its inhibition against inflammatory reaction， oxidative stress， and neuronal apoptosis and autophagy.

Recent studies have demonstrated that nitrite and ammonia levels are higher in the tumor environment, but their effects on cancer cells remains unclear. The present study was designed to determine the effects of nitrite and ammonia on tumor invasion and the role of reactive oxygen (ROS)/ornithine decarboxylase (ODC) pathway. SMMC-7721 cells were treated with sodium nitrite, ammonium chloride, sodium nitrite and ammonium chloride mixture for 24 h, the cell viability was analyzed using the MTT assay, cell invasion was analyzed with the transwell assay, the intracellular ROS levels were detected with a reactive oxygen species (ROS) test kits, the expression of intracellular ODC was examined with immunofluorescence and Western blot, the expression of matrix metallopeptidase-2(MMP-2) and MMP-9 were analyzed by Western blot. Compared with the control group, SMMC-7721 cells exhibited an increase in cell viability, invasion ability, ROS levels and ODC protein after exposure to 150 μmol·L^-1 sodium nitrite and ammonium chloride mixture for 24 h. The invasive activity was reduced by ROS scavenger N-acetycysteine (NAC) in SMMC-7721 cells. The specific ODC inhibitor difluoromethylornithine (DFMO) increased ROS levels and weakened the ability of sodium nitrite and ammonium chloride mixture in the regulation of invasion of SMMC-7721 cells. These data demonstrated that sodium nitrite and ammonium chloride mixture promote invasion of SMMC-7721 cells by enhancing ROS/ODC pathway.

Increasing evidence suggests that hepatocellular carcinomas (HCCs) are sustained by a distinct subpopulation of self-renewing cells known as cancer stem cells (CSC). However, our understanding of their regulation is limited. Rapid reversible changes of CSC-like cells within tumors may result from the effect of biological mediators found in the tumor microenvironment. This paper aims to explore how nitrite, a key cellular modulator whose level is elevated in many tumors, affects CSC-like phenotypes of human hepatoma cells SMMC-7721 cells. The SMMC-7721 cell line was cultured under serum-free conditions to produce floating spheres. The distribution of cell cycle was analyzed by flow cytometry, the capability of cells self-renew was detected by colony-forming capabilities and spheroid-formation assay, the expression of stemness protein such as CD133, CD90 and EpCAM were determined by flow cytometry and Western blot, cell invasion was analyzed by transwell assay, and viability of SMMC-7721 parental cells and spheroids cancer cells was determined by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Xenograft tumor models were established by subcutaneously injecting SMMC-7721 spheroids cancer cells, the transplanted tumor tissue ROS levels was detected by reactive oxygen species (ROS) test kits, the expression of HIF-1α was observed by immunofluorescence. Our results showed that the SMMC-7721 spheroid cells were enriched with CSCs properties, indicated by the ability to self-renew, increased expression of CSCs markers, and increased resistance to chemotherapeutic drugs. Additionally, SMMC-7721 parental cells and spheroids cancer cells were treated with 150 μmol·L^-1 sodium nitrite for 6 days, compared with control cells, an increased accumulation of G₀/G₁ phase cells was observable in treatment cells. Indeed, our data demonstrated that in parent cells and spheres cells that were treated with sodium nitrite for different time, the cells' ability to chemoresistance and invasion, clone-forming efficiencies and the spheres forming ability were significantly higher than that of control cells. Exposure of sodium nitrite regulated CSC-like phenotype, indicated by increased expression of known CSC markers, CD133, CD90 and EpCAM in the exposed parental cells, as well as in dormant spheroids cancer cells. Compared with the parent cells, the above effects of nitrite on the spheres cells were significantly enhanced. In vivo data also presented a more significant promotion of tumor xenograft growth from the nitrite treatment than from either of the control. Mechanistic analysis indicated that nitrite induced the upregulation of HIF-1α as well as the downregulation of ROS in the tumor microenvironment. These results suggest that nitrite increases the invasiveness of SMMC-7721 cells through up-regulation of tumor stemness.

In this study,liquiritigenin sulfonation was characterized using recombinant human sulfotransferases( SULTs). The chemical structure of liquiritigenin sulfate was determined by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS). Then model fitting and parameter estimation were performed using the Graphpad Prism V5 software. Various SULT enzymes( SULT1 A1,1 A2,1 A3,1 B1,1 C2,1 C4,1 E1 and 2 A1) were able to catalyze the formation of liquiritigenin-7-O-sulfate. Sulfonation of liquiritigenin-7-hydroxy( 7-OH) by these eight SULT enzymes consistently displayed the classical Michaelis-Menten profile. According to the intrinsic clearance( CLint) value,the sulfonation rates of liquiritigenin-7-OH by expressed SULT enzymes followed the following rank order: SULT1 C4 > SULT1 A3 > SULT1 E1 > SULT1 A1 > SULT1 A2 > SULT1 B1 >SULT1 C2>SULT2 A1. Further,liquiritigenin-7-O-sulfonation was significantly correlated with the SULT1 A3 protein levels( P<0. 05).Then,human embryonic kidney( HEK) 293 cells over expressing SULT1 A3( named as HEK-SULT1 A3 cells) were conducted. As a result,liquiritigenin-7-O-sulfate( L-7-S) was rapidly generated upon incubation of the cells with liquiritigenin. Consistent with SULT1 A3,sulfonation of liquiritigenin-7-OH in HEK-SULT1 A3 cells also followed the Michaelis-Menten kinetics. The derived Vmaxvalues was( 0. 315±0. 009) μmol·min-1·g-1,Kmwas( 7. 04±0. 680) μmol·L^-1,and CLintwas( 0. 045±0. 005) L·min^-1·g^-1. Moreover,the sulfonation characters of liquiritigenin( 7-OH) in SULT1 A3 were strongly correlated with that in HEK-SULT1 A3 cells( P<0. 001).The results indicated that HEK-SULT1 A3 cells have shown the catalytic function of SULT1 A3 enzymes. In conclusion,liquiritigenin was subjected to efficient sulfonation,and SULT1 A3 enzyme plays an important role in the sulfonation of liquiritigenin-7-OH. Significant sulfonation should be the main reason for the low bioavailability of liquiritigenin. In addition,HEK-SULT1 A3 cells were conducted and successfully used to evaluate liquiritigenin sulfonation,which will provide an appropriate tool to accurately depict the sulfonation disposition of liquiritigenin in vivo.
Arylsulfotransferase ; Flavanones/metabolism* ; Humans ; Tandem Mass Spectrometry

Anti-Inflammatory Agents ; therapeutic use ; Antioxidants ; therapeutic use ; Diabetes Mellitus, Type 2 ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Herbal Medicine ; methods ; Humans ; Hypoglycemic Agents ; therapeutic use

The p26, a multifunctional ubiquitin-binding protein, has been proposed to be involved in protein degradation as a component within the ubiquitin-proteasome and autophagy-lysosome systems. As a scaffolding protein with several different kinds of protein-protein interaction domains, p62 mediates various cellular functions. Importantly, p62 plays a critical role in cell's selective autophagy and oxidative stress response, which are associated with the pathogenesis of several human diseases. In this review, we describe the structure of p62 and the mechanism of connection between p62 and ubiquitin-proteasome system/autophagy, so as to provide some perspectives on p62 research.
Adaptor Proteins, Signal Transducing ; physiology ; Autophagy ; Humans ; Oxidative Stress ; Proteasome Endopeptidase Complex ; Protein Interaction Domains and Motifs ; Proteolysis ; Ubiquitin

This study was designed to explore the effect of apigenin (Api) on dendritic cell (DCs) maturation and function in murine spleen cells. The single spleen cell was isolated, and then cultured with lipopolysaccharide (LPS) in the present and absence of apigenin. After 24 h, the toxicity of Api and the T cell proliferation were determined by CCK8 kit. In addition, we collected the cell-free supernatants to measure cytokine production using ELISA, collected the cells to determine the DC maturation using flow cytometry. Finally, we purified Api and/or LPS-treated CD11c⁺ DCs which were pulsed with ovalbumin (OVA)_323-339 and then were adoptive transferred into C57BL/6 mice to detect the OVA_323-339-specific T cell proliferation and T helper (Th1) and Th2 cell secreting IFN-γ and IL-4 production, respectively. We found that Api did not affect splenocyte viability, but inhibited the production of pro-inflammatory cytokine IL-1β, IL-6 and TNF-α, not anti-inflammatory cytokine IL-10. In addition, Api inhibited the expression of co-stimulatory CD80, CD86 and MHCII of CD11c⁺ DCs. Finally, compared to LPS+OVA DCs group, DCs from Api and LPS co-treated splenocytes (Api+LPS+DCs) impaired OVA_323-339-specific T cell proliferation and the production of IFN-γ and IL-4 in CD4⁺ T cells, which had the similar responses with OVA+DCs. These data suggest that Api exhibits anti-inflammatory properties via inhibiting DC activation and function, as a new immune-modulator, which may induce immune-tolerance with a benefit to those with chronic inflammation.

Objective:To observe the efficacy of modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation), and investigate its effects on serum levels of intestinal neurotransmitter nitric oxide synthase (nNOS), nitric oxide (NO), and vasoactive intestinal peptide (VIP), as well as superoxide dismutase (SOD), malonaldehyde (MDA), and glutathione (GSH) levels. Method:One hundred and ten patients with functional constipation were selected and randomly divided into control group (55 cases) and treatment group (55 cases) by referring to random number table. The patients in control group were given with routine therapy, Domperidone tablet (1 tablet/time, tid), and Phenolphthalein tablets (100 mg/time, bid). The patients in treatment group were treated with modified Simotang, 1 dose/day. Both groups were treated for 4 weeks. Then the scores of main clinical symptoms of functional constipation, scores of Qi-stagnancy constipation and clinical efficacy were compared between two groups. Constipation recurrence rate was compared between two groups after stopping medicine. Serum levels of intestinal neurotransmitters nNOS, NO and VIP as well as SOD, MDA, GSH levels were detected in both groups. Result:After treatment, scores for main clinical symptoms (difficult defecation, abdominal distension, defecation time, number of defecation times) and Bristol scores in treatment group were obviously lower than those in control group (P<0.01). Scores for symptoms of Qi-stagnancy constipation (ungratifying defecation, abdominal distension,bowel ringing, frequent flatus, chest and flank tightness) in treatment group were obviously lower than those in control group after treatment (P<0.01). Total clinical efficacy in treatment group 98.04% was superior to that in control group 84.62% ( P<0.05). Constipation recurrence rate was 3.92% and 8.16% in treatment group after 4 and 8 weeks of medication stopping, obviously lower than those in control group 18.18% and 27.78% (P<0.05). After treatment, serum levels of intestinal neurotransmitters nNOS, NO, VIP in treatment group were remarkably lower than those in control group (P<0.01). After treatment, serum levels of SOD and GSH in treatment group were higher while MDA level was lower than those in control group (P<0.01). Conclusion:Based on routine therapy, modified Simotang in treatment of adult functional constipation (Qi-stagnancy constipation) can improve clinical symptoms and syndrome symptoms, increase the clinical efficacy, decrease recurrence rate, and regulate levels of intestinal neurotransmitters nNOS,NO and VIP as well as SOD, MDA, GSH levels.