Dual Regulation of Nicotine on NLRP3 Inflammasome in Macrophages with the Involvement of Lysosomal Destabilization, ROS and α7nAChR.

Nicotine, the primary alkaloid in tobacco products, has been shown to have immunoregulatory function in at least 20 diseases. The biological mechanism of action of nicotine immunoregulation is complex, resulting in an improvement of some disease states and exacerbation of others. Given the central role of the NLRP3 inflammasome in macrophages among multiple inflammatory diseases, this study examined how nicotine alters NLRP3 inflammasome activation in macrophages. NLRP3 inflammasome activation was examined mechanistically in the context of different nicotine dosages. We show NLRP3 inflammasome activation, apoptosis-associated speck-like protein (ASC) expression, caspase-1 activity and subsequent IL-1ß secretion were positively correlated with nicotine in a dose-dependent relationship, and destabilization of lysosomes and ROS production were also involved. At high concentrations of nicotine surpassing 0.25 mM, NLRP3 inflammasome activity declined, along with increased expression of the anti-inflammatory Alpha7 nicotinic acetylcholine receptor (α7nAChR) and the inhibition of TLR4/NF-κB signaling. Consequently, high doses of nicotine also reduced ASC expression, caspase-1 activity and IL-1ß secretion in macrophages. Collectively, these results suggest a dual regulatory function of nicotine on NLRP3 inflammasome activation in macrophages, that is involved with the pro-inflammatory effects of lysosomal destabilization and ROS production. We also show nicotine mediates anti-inflammatory effects by activating α7nAChR at high doses.

Efficacy of radiofrequency ablation combined with sorafenib for treating liver cancer complicated with portal hypertension and prognostic factors.

BACKGROUND: Patients with liver cancer complicated by portal hypertension present complex challenges in treatment. AIM: To evaluate the efficacy of radiofrequency ablation in combination with sorafenib for improving liver function and its impact on the prognosis of patients with this condition. METHODS: Data from 100 patients with liver cancer complicated with portal hypertension from May 2014 to March 2019 were analyzed and divided into a study group (n = 50) and a control group (n = 50) according to the treatment regimen. The research group received radiofrequency ablation (RFA) in combination with sorafenib, and the control group only received RFA. The short-term efficacy of both the research and control groups was observed. Liver function and portal hypertension were compared before and after treatment. Alpha-fetoprotein (AFP), glypican-3 (GPC-3), and AFP-L3 levels were compared between the two groups prior to and after treatment. The occurrence of adverse reactions in both groups was observed. The 3-year survival rate was compared between the two groups. Basic data were compared between the survival and non-surviving groups. To identify the independent risk factors for poor prognosis in patients with liver cancer complicated by portal hypertension, multivariate logistic regression analysis was employed. RESULTS: When comparing the two groups, the research group's total effective rate (82.00%) was significantly greater than that of the control group (56.00%; P < 0.05). Following treatment, alanine aminotransferase and aspartate aminotransferase levels increased, and portal vein pressure decreased in both groups. The degree of improvement for every index was substantially greater in the research group than in the control group (P < 0.05). Following treatment, the AFP, GPC-3, and AFP-L3 levels in both groups decreased, with the research group having significantly lower levels than the control group (P < 0.05). The incidence of diarrhea, rash, nausea and vomiting, and fatigue in the research group was significantly greater than that in the control group (P < 0.05). The 1-, 2-, and 3-year survival rates of the research group (94.00%, 84.00%, and 72.00%, respectively) were significantly greater than those of the control group (80.00%, 64.00%, and 40.00%, respectively; P < 0.05). Significant differences were observed between the survival group and the non-surviving group in terms of Child-Pugh grade, history of hepatitis, number of tumors, tumor size, use of sorafenib, stage of liver cancer, histological differentiation, history of splenectomy and other basic data (P < 0.05). Logistic regression analysis demonstrated that high Child-Pugh grade, tumor size (6-10 cm), history of hepatitis, no use of sorafenib, liver cancer stage IIIC, and previous splenectomy were independent risk factors for poor prognosis in patients with liver cancer complicated with portal hypertension (P < 0.05). CONCLUSION: Patients suffering from liver cancer complicated by portal hypertension benefit from the combination of RFA and sorafenib therapy because it effectively restores liver function and increases survival rates. The prognosis of patients suffering from liver cancer complicated by portal hypertension is strongly associated with factors such as high Child-Pugh grade, tumor size (6-10 cm), history of hepatitis, lack of sorafenib use, liver cancer at stage IIIC, and prior splenectomy.

The Role of Nicotine Metabolic Rate on Nicotine Dependence and Rewarding: Nicotine Metabolism in Chinese Male Smokers and Male Mice.

The exact relationship between nicotine metabolism and dependence is not fully understood but is known to be influenced at a molecular level by genetic factors. A sample comprising 274 Chinese adult male smokers was categorized into groups based on their metabolic rates, namely fast, intermediate, and slow metabolizers. We then measured their smoking topography, evaluated their nicotine dependence, and assessed the rewarding effects. Based on these findings, we proposed the hypothesis that the rate of nicotine metabolism could influence the level of dopamine release which in turn had repercussions on the pleasurable and rewarding effects. To test this hypothesis, male mice were selected with different nicotine metabolic rates that closely resembled in the smoker group. We evaluated their nicotine dependence and rewarding effects through conditioned place preference and withdrawal symptom tests, supplemented with dopamine release measurements. In both animal and human, the slow metabolism group (SMG) required less nicotine to maintain a comparable level of dependence than the fast metabolism group (FMG). The SMG could achieve similar rewarding effects to FMG despite consuming less nicotine. Comparable dopamine levels released were therefore critical in setting the nicotine acquisition behavior in this animal model and also for the smokers tested. Our findings suggested that even within the same ethnicity of established smokers (Chinese Han), differences in nicotine metabolism were an important parameter to modulate the degree of nicotine dependence.

Cited2 inhibited hypoxia-induced proliferation and migration of PASMCs via the TGF-ß1/Cited2/PPARγ pathway.

Objectives: Proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) contribute to hypoxia-induced pulmonary hypertension (HPH). The transcription factor Cbp/p300-interacting transactivator with Glu/Asp-rich carboxy-terminal domain 2 (Cited2) has been implicated in the control of tumor cells and mesenchymal stem cell (MSC) and cardiomyocyte growth or migration. Whether Cited2 is involved in the proliferation and migration of PASMCs and the underlying mechanisms deserve to be explored. Materials and Methods: Cited2 expression was detected in rat PASMCs under hypoxia conditions and HPH rat models. The effect of Cited2 on the proliferation and migration of PASMC was detected by overexpression or knockdown of the Cited2 gene. After PAMSCs were treated with recombinant TGF-ß1 and the lentivirus vector overexpressing Cited2, expression of peroxisome proliferator-activated receptor gamma (PPARγ) was examined by western blotting. Results: We revealed that hypoxia down-regulated the expression of Cited2 in PASMCs and rat pulmonary arteries. Cited2 overexpression inhibited the proliferation and migration of PASMCs under hypoxia, while Cited2 knockdown induced the proliferation and migration of PASMCs. Cited2 inhibits the negative regulation of the TGF-ß1 pathway on PPARγ to inhibit the proliferation and migration of PASMCs. Conclusion: These findings suggest that increased Cited2 expression contributes to the inhibition of PASMCs proliferation and migration by regulating TGF-ß1-mediated target gene expression in HPH and provides a new target for molecular therapy of HPH.

A comprehensive metabolomics analysis of volatile and non-volatile compounds in matcha processed from different tea varieties.

Tea varieties play a crucial role on the quality formation of matcha. This research aimed to examine the impact of four specific tea plant varieties (Okumidori, Longjing 43, Zhongcha108, and E'Cha 1) on various aspects of matcha, including sensory evaluation, major components, color quality, volatile and non-volatile metabolomic profiles. The findings revealed that the levels of tea polyphenols, ester catechins, nonester catechins, and amino acids varied among these four varieties. Notably, 177 significant different metabolites, such as phenolic acids, flavonoids, tannins, alkaloids were identified among 1383 non-volatile compounds. In addition, 97 key aroma-active compounds were identified based on their odor activity value exceeding 1. Aldehydes, heterocyclic compounds, and ketones were closely associated with the formation of volatile metabolites. Overall, this study enhances our understanding of how different tea plant varieties impact the quality of matcha, and can provide valuable guidance for improving matcha varieties in a favorable direction.

Phylogenetic and Taxonomic Analyses Reveal Three New Wood-Inhabiting Fungi (Polyporales, Basidiomycota) in China.

Three new wood-inhabiting fungal species, Cerioporus yunnanensis, Perenniporiopsis sinensis, and Sarcoporia yunnanensis, are proposed based on a combination of the morphological features and molecular evidence. Cerioporus yunnanensis is characterized by the pileate basidiomata having a fawn brown to black pileal surface, a dimitic hyphal system with clamped generative hyphae, and the presence of the fusoid cystidioles and cylindrical basidiospores (9-12.5 × 3.5-5 µm). Perenniporiopsis sinensis is distinct from the osseous pileus with verrucose, an orange-yellow to dark reddish-brown pileal surface with a cream margin, a trimitic hyphal system with clamped generative hyphae, and the presence of the fusiform cystidioles and ellipsoid basidiospores (9-11 × 5.5-6.5 µm). Sarcoporia yunnanensis is typical of the pileate basidiomata with a salmon to reddish-brown pileal surface, a monomitic hyphal system with clamped generative hyphae, and the presence of the ellipsoid basidiospores (4-5.5 × 2.5-4 µm). Sequences of ITS + nLSU + mt-SSU + TEF1 + RPB1 + RPB2 genes were used for the phylogenetic analyses using maximum likelihood, maximum parsimony, and Bayesian inference methods. The multiple genes with six loci analysis showed that the three new species nested within the order Polyporales, in which C. yunnanensis and P. sinensis nested into the family Polyporaceae, and S. yunnanensis grouped into the family Sarcoporiaceae.

Liver toxicity in rats after subchronic exposure to HTP aerosol and cigarette smoke.

Background: Heated tobacco product (HTP) considered to be a novel tobacco product which was reported safer than traditional cigarettes evidenced by lower potential harmful components released. Liver is an important detoxification organ of the body, the chemical components in aerosols are metabolized in the liver after absorbed, so it is necessary to explore the effect of HTP on the liver. Materials and Methods: The potential effect of HTP and cigarette smoke (CS) on SD rats was explored according to OECD 413 subchronic inhalation. The rats were randomly divided into Sham (air), different dosage of HTP groups (HTP_10, 23 and 50 µg nicotine/L aerosol) and Cig_23 (23 µg nicotine/L aerosol) group. After exposure, the clinical pathology, inflammation and oxidative stress were measured. Results: The clinical pathology results showed that both HTP_50 and Cig_23 led to abnormality of ALT for male rats. CS and HTP exposure reduced the expression of IL-1ß, IL-6 and TNF-α and mitochondrial medicated oxidative stress. In addition, the ATP production was reduced in Cig_23 group. Although inflammation and oxidative stress were displayed, no apoptosis were observed by TUNEL assay and these existed obvious pathological changes only in HTP_50 group, while in CS group with equivalent nicotine, hepatocytes swelling were observed in liver. Conclusion: CS exposure induced liver damage through mitochondrial mediated oxidative stress and inflammation, which was also observed in high concentration of HTP exposure group. For the same equivalent nicotine, HTP may show lower toxic effect on liver than CS.

Cell death induced by nicotine in human neuroblastoma SH-SY5Y cells is mainly attributed to cytoplasmic vacuolation originating from the trans-Golgi network.

Humans are usually exposed to nicotine through the use of tobacco products. Although it is generally believed that nicotine is relatively harmless in tobacco consumption, it is, in fact, a toxic substance that warrants careful consideration of its potential toxicity. However, the current understanding of the neurotoxicity of nicotine is still very limited. In this study, we aim to reveal the toxic risk of nicotine to key target neuronal cells and its potential toxic mechanisms. The results showed that nicotine induced cell death, ROS increase, mitochondrial membrane potential decrease, and DNA damage in SH-SY5Y human neuroblastoma cells at millimolar concentrations, but did not cause toxic effects at the physiological concentration. These toxic effects were accompanied by cytoplasmic vacuolation. The inhibition of cytoplasmic vacuolation by bafilomycin A1 greatly reduced nicotine-induced cell death, indicating that cytoplasmic vacuolation is the key driving factor of cell death. These cytoplasmic vacuoles originated from the trans-Golgi network (TGN) and expressed microtubule-associated protein 1 light chain 3-II (LC3-II) and lysosomal associated membrane protein 1(LAMP1). The presence of LC3-II and LAMP1 within these vacuoles serves as evidence of compromised TGN structure and function. These findings provide valuable new insights into the potential neurotoxic risk and mechanisms of nicotine.

Evaluation of toxicity of heated tobacco products aerosol and cigarette smoke to BEAS-2B cells based on 3D biomimetic chip model.

It is still a controversial topic about evaluating whether heated tobacco products (HTP) really reduce harm, which involves the choice of an experimental model. Here, a three-dimensional (3D) biomimetic chip model was used to evaluate the toxicity of aerosols came from HTP and smoke produced by cigarettes (Cig). Based on cell-related experiments, we found that the toxicity of Cig smoke extract diluted four times was also much higher than that of undiluted HTP, showing higher oxidative stress response and cause mitochondrial dysfunction. Meanwhile, both tobacco products all affect the tricarboxylic acid cycle (TCA), which is manifested by a significant decrease in the mRNA expression of TCA key rate-limiting enzymes. Summarily, 3D Biomimetic chip technology can be used as an ideal model to evaluate HTP. It can provide important data for tobacco risk assessment when 3D chip model was used. Our experimental results showed that HTP may be less harmful than tobacco cigarettes, but it does show significant cytotoxicity with the increase of dose. Therefore, the potential clinical effects of HTP on targeted organs such as lung should be further studied.

Gliclazide Reduces Colitis-Associated Colorectal Cancer Formation by Deceasing Colonic Inflammation and Regulating AMPK-NF-κB Signaling Pathway.

BACKGROUND: Gliclazide is a potential anti-cancer drug candidate for preventing carcinogenesis. However, the effect of gliclazide on colitis-associated colorectal cancer remains unknown. AIMS: We aimed to evaluate whether gliclazide plays a protective role in colitis-associated colorectal cancer and the underlying molecular mechanism. METHODS: The administration of azoxymethane (AOM) followed by dextran sulfate sodium (DSS) aimed to induce colitis-associated colorectal cancer in mice. C57BL mice were gavaged with gliclazide (6 mg/kg by gavage 5 days a week) for 12 weeks immediately following AOM administration. After sacrificing the mice, colon tissues were measured for tumor number and tumor burden. The proliferation- and inflammation-related molecular mechanisms were explored. RESULTS: The administration of gliclazide significantly reduced the tumor number and tumor burden in mice. Cell proliferation decreased in the gliclazide group compared with the control group, as indicated by reduced Ki-67 expression. Furthermore, gliclazide alleviated colonic inflammation, significantly decreased pro-inflammatory factor TNF-α levels and increased anti-inflammatory factor IL-10 levels in vivo. In vivo and vitro, it was shown that gliclazide increased the level of phospho-AMPK (p-AMPK) and inhibited NF-κB activity. Further studies demonstrated that the inhibition of NF-κB activity induced by gliclazide was mediated by p-AMPK in vitro. CONCLUSIONS: Gliclazide effectively alleviated colonic inflammation and prevented colonic carcinogenesis in an AOM-DSS mouse model by modulating the AMPK-NF-κB signaling pathway. Thus, gliclazide holds potential as a chemopreventive agent for colitis-associated colorectal cancer.

Testicular tissue response following a 90-day subchronic exposure to HTP aerosols and cigarette smoke in rats.

Background: Researches have shown that chronic inhalation of cigarette smoke (CS) disrupts male reproductive system, but it is unclear about the mechanisms behind reproductive damages by tobacco toxicants in male rats. This study was designed to explore the effects of heated tobacco products (HTP) aerosols and CS exposure on the testicular health of rats. Materials and Methods: Experiments were performed on male SD rats exposed to filtered air, HTP aerosols at 10 µg/L, 23 µg/L, and 50 µg/L nicotine-equivalent contents, and also CS at 23 µg/L nicotine-equivalent content for 90 days in five exposure groups (coded as sham, HTP_10, HTP_23, HTP_50 and Cig_23). The expression of serum testosterone, testicular tissue inflammatory cytokines (IL-1ß, IL-6, IL-10, TNF-α), reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA), NLRP3 inflammasome-related mRNAs and proteins (NLRP3, ASC, and Caspase-1), the degree of pyroptosis and histopathology were investigated. Results: The results demonstrated that HTP_50 and Cig_23 caused varying degrees of oxidative damage to rat testis, resulting in a decrease of sperm quantity and serum testosterone contents, an increase in the deformity rate, expression levels of proinflammatory cytokines, and NLRP3 inflammasome-related mRNA, and an increase in the NLRP3, ASC, and Caspase-1-immunopositive cells, pyroptosis cell indices, and histopathological damage in the testes of rats. Responses from the HTP_10 and HTP_23 groups were less than those found in the above two exposure groups. Conclusion: These findings indicate that HTP_50 and Cig_23 induced oxidative stress in rat testes, induced inflammation and pyroptosis through the ROS/NLRP3/Caspase-1 pathway, and destroyed the integrity of thetesticular tissue structure.

SIRT3 Regulates the ROS-FPR1/HIF-1α Axis under Hypoxic Conditions to Influence Lung Cancer Progression.

Hypoxia-inducible factor (HIF-1α) is a therapeutic target in lung cancer, and the deacetylase sirtuin 3 (SIRT3) is closely associated with tumorigenesis. Formyl peptide receptor 1 (FPR1) is involved in a wide range of physiopathological processes in various tumor cells. We explored whether SIRT3 affects the development of lung cancer by regulating the reactive oxygen species (ROS)-FPR1/HIF-1α axis under hypoxic conditions. The effects of SIRT3 overexpression on the levels of FPR1, HIF-1α, ROS, inflammatory factors, and cell proliferation and migration in A549 cells under hypoxic conditions were assessed in combination with the FPR1 inhibitor. BALB/c nude mice were subcutaneously injected with cancer cells transfected/untransfected with SIRT3 overexpressing lentiviral vectors. Immunohistochemistry and enzyme-linked immunosorbent assay were performed to detect SIRT3 expression and the expression levels of IL-1ß, TNF-α, and IL-6, respectively, in tumor tissues. Cell proliferation, invasion, migration, and IL-1ß, TNF-α, IL-6, and ROS levels were significantly higher in the Hypoxia group than in the Control group. Moreover, the mRNA and protein expression levels of SIRT3 were significantly down-regulated, whereas they were significantly up-regulated for FPR1 and HIF-1α. In contrast, SIRT3 overexpression in a hypoxic environment inhibited cell proliferation, invasion, and migration, decreased IL-1ß, TNF-α, IL-6, and ROS levels, up-regulated the mRNA and protein expression levels of SIRT3, and down-regulated the mRNA and protein expression levels of FPR1 and HIF-1α. In addition, we found the same results in tumorigenic experiments in nude mice. SIRT3 in hypoxic environments may affect tumor cell proliferation, invasion, migration, and inflammation levels via the ROS-FPR1/HIF-1α axis, thereby inhibiting tumor cell development.

Generation of an iPSC line (TSHSUi001-A) from a patient with Peutz-Jeghers syndrome due to STK11 mutation.

We established an iPSC line (TSHSUi001-A) from a patient with Peutz-Jeghers syndrome, carrying heterozygous c.290 + 1G > A mutation in STK11 gene. Peripheral blood mononuclear cells were reprogrammed using non-integrating delivery of OCT4, SOX2, KFL4, BCL-XL and c-MYC. The iPSC line expressed pluripotency markers, could differentiate into cells of three germ layers in vitro, and displayed a normal karyotype.

Acetaldehyde induces similar cytotoxic and genotoxic risks in BEAS-2B cells and HHSteCs: involvement of differential regulation of MAPK/ERK and PI3K/AKT pathways.

Long-term use of alcohol and cigarettes is associated with millions of deaths each year, directly or indirectly. The carcinogen acetaldehyde is both a metabolite of alcohol and the most abundant carbonyl compound in cigarette smoke, and co-exposure of them is usual and primarily leads to liver and lung injury, respectively. However, few studies have explored the synchronic risk of acetaldehyde on the liver and lung. Here, we investigated the toxic effects and related mechanisms of acetaldehyde based on normal hepatocytes and lung cells. The results showed that acetaldehyde caused significant dose-dependent increases of cytotoxicity, ROS level, DNA adduct level, DNA single/double-strand breakage, and chromosomal damage in BEAS-2B cells and HHSteCs, with similar effects at the same doses. The gene and protein expression and phosphorylation of p38MAPK, ERK, PI3K, and AKT, key proteins of MAPK/ERK and PI3K/AKT pathways regulating cell survival and tumorigenesis, were significantly upregulated on BEAS-2B cells, while only protein expression and phosphorylation of ERK were upregulated significantly, the other three decreased in HHSteCs. When either the inhibitor of the four key proteins was co-treated with acetaldehyde, cell viabilities were almost unchanged in BEAS-2B cells and HHSteCs. Thus, acetaldehyde could synchronically induce similar toxic effects in BEAS-2B cells and HHSteCs, and MAPK/ERK and PI3K/AKT pathways seem to be involved in different regulatory mechanisms.

Propensity score matching analysis of efficacy of drug-eluting beads (DEB)-TACE loaded with different drugs in the treatment of hepatocellular carcinoma.

OBJECTIVE: To evaluate the efficacy of drug-eluting beads transarterial chemoembolization (DEB-TACE) in the treatment of hepatocellular carcinoma (HCC) by propensity score matching (PSM) technique. METHODS: The clinical data of HCC patients treated with DEB-TACE in the First Affiliated Hospital of Zhengzhou University from June 2017 to June 2020 as well as their 36-month-follow-up data were retrospectively analyzed. The subjects were matched in pairs based on baseline data and laboratory indicators using the PSM method and divided into a pirarubicin group (n = 34), raltitrexed group (n = 34), and arsenic trioxide group (n = 34). Clinical efficacy was evaluated according to mRECIST criteria. The levels of alpha fetal protein (AFP), carcinoma embryonic antigen (CEA) and carbohydrate antigen-125 (CA125) in serum were detected by enzyme-linked immunosorbent assay (ELISA). The progression-free survival (PFS) and overall survival (OS) were recorded by outpatient, inpatient, and telephone follow-up. Adverse reactions were counted. RESULTS: After PSM, no significant differences were seen in gender, age, tumor burden, Child-Pugh grade, portal vein tumor thrombus or TACE frequency among the three groups (all P>0.05). The ORR rate of the pirarubicin group and arsenic trioxide group at both 3rd and 6th month post-operation was significantly higher than that of the raltitrexed group (all P<0.05). Before and 1 month after treatment, there were no significant differences in the aspartate aminotransferase (AST), alanine aminotransferase (ALT), or total bilirubin (TBIL) levels between the three groups (all P>0.05). Before treatment, no significant differences were observed in AFP, CEA, or CA125 levels among the three groups (all P>0.05). After treatment, the levels of AFP in the pirarubicin group and arsenic trioxide group were lower than those in the raltitrexed group (both P<0.05), but there were no significant differences in CEA and CA125 levels (all P>0.05). There were no significant differences in PFS and OS among the three groups (all P>0.05), and the incidence of fever, abdominal pain, and myelosuppression showed no significant differences among the three groups (all P>0.05). CONCLUSION: The efficacies of DEB-TACE loaded with pirarubicin, raltitrexed, or arsenic trioxide in treating HCC were generally comparable, and the survival benefit of patients was similar. The short-term efficacy of the pirarubicin group and arsenic trioxide group was slightly better than that of the raltitrexed group.

Protective effects of isorhamnetin against H2O2-induced oxidative damage in HaCaT cells and comprehensive analysis of key genes.

Isorhamnetin (ISO) is a methylated flavonol present in the leaves, flowers, and fruits of many plants with antitumour, anti-inflammatory, antioxidant, and anti-apoptotic properties. ISO has been suggested as the active substance in Vernonia anthelmintica (L.) to treat vitiligo. However, the mechanisms underlying its effects remain unclear. In this study, human keratinocytes (HaCaT cells) were pre-treated with or without ISO and then stimulated with hydrogen peroxide (H2O2) to generate oxidative damage. Pre-treatment with ISO increased HaCaT cell viability, reduced malondialdehyde content, and enhanced superoxide dismutase activity, resulting in a reduction in the loss of mitochondrial membrane potential, improved cell morphological damage, and apoptosis inhibition. Furthermore, we identified 51 significantly dysregulated differentially expressed genes (DEGs) of HaCaT cells treated with ISO using RNA-sequencing. Enrichment analysis using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases indicated that the protective effect of ISO could be related to its effects on the Wnt signalling pathway. Our study provides novel insights into key gene regulation in the progression of oxidative damage and the mechanisms of action of ISO.

Evaluation of oil accumulation and biodiesel property of Lindera glauca fruits among different germplasms and revelation of high oil producing mechanism for developing biodiesel.

BACKGROUND: Lindera glauca with rich resource and fruit oil has emerged as novel source of biodiesel in China, but different germplasms show a variation for fruit oil content and FA profile. To develop L. glauca fruit oils as biodiesel, a concurrent exploration of oil content, FA composition, biodiesel yield, fuel property and prediction model construction was conducted on the fruits from 8 plus germplasms to select superior genotype for ideal biodiesel production. Another vital focus was to highlight mechanism that govern the differences in oil content and FA profile of different germplasms. The cross-accessions comparisons associated with oil-synthesized gene transcriptional level and oil accumulative amount led to the identification of potential determinants (enzymes, transporters or transcription factors) and regulatory mechanisms responsible for high-quality oil accumulation. RESULTS: To select superior germplasm and unravel regulatory mechanism of high oil production for developing L. glauca fruit oils as biodiesel, 8 plus trees (accession LG01/02/03/04/05/06/07/08) with high-yield fruits were selected to evaluate the differences in oil content, FA profile, biodiesel yield and fuel property, and to construct fuel property prediction model, revealing a variation in the levels of fruit oil (45.12-60.95%), monounsaturated FA (52.43-78.46%) and polyunsaturated FA (17.69-38.73%), and biodiesel yield (80.12-98.71%) across different accessions. Of note, LG06 had a maximum yield of oil (60.95%) and biodiesel (98.71%), and ideal proportions of C18:1 (77.89%), C18:2 (14.16%) and C18:3 (1.55%), indicating that fruit oils from accession LG06 was the most suitable for high-quality biodiesel production. To highlight molecular mechanism that govern such differences in oil content and FA composition of different accessions, the quantitative relationship between oil-synthesized gene transcription and oil accumulative amount were conducted on different accessions to identify some vital determinants (enzymes, transporters or transcription factors) with a model of carbon metabolic regulatory for high-quality oil accumulation by an integrated analysis of our recent transcriptome data and qRT-PCR detection. Our findings may present strategies for developing L. glauca fruit oils as biodiesel feedstock and engineering its oil accumulation. CONCLUSIONS: This is the first report on the cross-accessions evaluations of L. glauca fruit oils to determine ideal accession for producing ideal biodiesel, and the associations of oil accumulative amount with oil-synthesized gene transcription was performed to identify some crucial determinants (enzymes, transporters or transcription factors) with metabolic regulation model established for governing high oil production. Our finding may provide molecular basis for new strategies of developing biodiesel resource and engineering oil accumulation.

Pharmacokinetics of freebase nicotine and nicotine salts following subcutaneous administration in male rats.

Nicotine lactate, nicotine tartrate, nicotine benzoate, and freebase nicotine (FBN) are four forms of nicotine salt systems that are present in tobacco products. However, few in vivo studies have compared their pharmacological (pK) efficacies, which are important for understanding their roles in the addiction and abuse of tobacco and nicotine products. In this work, the pK of the above nicotine salt systems was studied by subcutaneously injecting their aqueous solutions in rats and obtaining blood samples from the jugular vein. Nicotine levels in the blood were analyzed by LC-MS/MS. The results demonstrated that rapid nicotine absorption occurred in all nicotine systems. Of them, NB had the smallest Tmax , while FBN had the largest Tmax . The nicotine metabolic rate and clearance decreased for FBN, indicating that nicotine retention in the body was higher than for the other three salt-based systems. Compared with nicotine salts, FBN could reach and maintain a higher concentration in the animal model. Additionally, as the benzoic acid ratios increased, the Cmax of the nicotine benzoate (NB) in the plasma decreased. This indicates that the lower the pH, the lower the Cmax . When different concentrations of NB were used, the higher the NB concentration, the greater the Cmax and AUC(0-t) . These results demonstrate that nicotine adsorption by NB in the animal model depended on both pH and concentration. This baseline information could be used to explain different clinical pharmacological observations in humans, though this study only considered the effects of nicotine on pharmacokinetics in vivo.

[Analysis of characteristics of the location of lower extremity deep vein thrombosis and the site of pulmonary embolism in hospitalized patients].

OBJECTIVE: To investigate the characteristics and relationship between the location of lower extremity deep vein thrombosis (DVT) and the site of pulmonary embolism in hospitalized patients. METHODS: The data of patients with lower extremity DVT diagnosed by ultrasound examination and pulmonary embolism diagnosed by CT pulmonary angiography from December 2017 to December 2021 were analyzed retrospectively. According to the location of lower extremity DVT, the patients were divided into mixed DVT, proximal DVT, and distal DVT which was further divided into anterior/posterior tibial vein or peroneal vein thrombosis and calf muscular venous thrombosis. Mixed DVT was referred to the presence of both proximal and distal DVT. According to the involved site of pulmonary artery, pulmonary embolism was divided into three types: main pulmonary artery, left or right pulmonary artery trunk embolism, lobar pulmonary artery embolism and segmental pulmonary artery embolism. The location of lower extremity DVT, the site of pulmonary embolism, the clinical manifestation (shortness of breath, chest tightness, chest pain, hemoptysis, cough, lower limb swelling, lower limb pain, syncope, fever) and risk factors (fracture/trauma, tumor, diabetes, hypertension, atrial fibrillation, infection, surgery, autoimmune diseases, paralysis, pregnancy) of venous thromboembolism (VTE), and the level of D-dimer were analyzed. RESULTS: A total of 209 patients were enrolled finally, including 127 patients with left lower extremity DVT (60.8%) and 82 with right lower extremity DVT (39.2%). Mixed DVT accounted for 39.2%, proximal DVT accounted for 17.3%, and distal DVT accounted for 43.5% (anterior/posterior tibial vein and peroneal vein thrombosis accounted for 14.8%, calf muscular venous thrombosis accounted for 28.7%). The incidences of main pulmonary artery embolism, left or right pulmonary artery trunk embolism in the mixed DVT and proximal DVT were significantly higher than those in the anterior/posterior tibial vein or peroneal vein thrombosis and calf muscular venous thrombosis [41.5% (34/82), 38.8% (14/36) vs. 16.2% (5/31), 10.0% (6/60)], with statistically significant differences (all P < 0.05). The incidences of pulmonary segmental artery embolism in the anterior/posterior tibial vein or peroneal vein thrombosis were higher than those in the mixed DVT and proximal DVT [41.9% (13/31) vs. 26.8% (22/82), 30.6% (11/36)], but the difference was not statistically significant (both P > 0.05). The incidences of pulmonary segmental artery embolism in the calf muscular venous thrombosis were significantly higher than those in the mixed DVT and the proximal DVT [66.7% (40/60) vs. 26.8% (22/82), 30.6% (11/36)], and the difference was statistically significant (both P < 0.05). The levels of D-dimer in patients with calf muscular venous thrombosis combined with main pulmonary artery embolism, left or right pulmonary artery trunk embolism were significantly higher than those in patients with calf muscular venous thrombosis combined pulmonary segmental artery embolism (mg/L: 6.08±3.12 vs. 3.66±2.66, P < 0.05). There were no significant differences in D-dimer levels in other patients with DVT combined with pulmonary embolism in different sites. In terms of the clinical manifestations of VTE, the incidences of lower limb swelling in the mixed DVT and proximal DVT were significantly higher than those in the anterior/posterior tibial vein or peroneal vein thrombosis and calf muscular venous thrombosis [54.9% (45/82), vs. 29.0% (9/31), 15.0% (9/60), both P < 0.05], the incidences of lower limb swelling in the proximal DVT were significantly higher than those in the calf muscular venous thrombosis [41.7% (15/63) vs. 15.0% (9/60), P < 0.05], there were no significant difference in the other clinical manifestations among the DVT groups. There was no significant difference in the incidence of VTE risk factors among the groups. CONCLUSIONS: The DVT of inpatients mostly occurred in the left lower limb, and the incidence of distal DVT was higher than that of proximal DVT. Mixed DVT and proximal DVT combined with pulmonary embolism mostly occurred in the main pulmonary artery, left or right pulmonary artery trunk, while distal DVT combined with pulmonary embolism mostly occurred in the pulmonary segmental artery. The levels of D-dimer in patients with lower extremity DVT combined with main pulmonary artery or left and right pulmonary artery trunk embolism were higher than those in patients with pulmonary lobe and segmental artery embolism. The incidence of lower extremity swelling in patients with mixed DVT and proximal DVT was higher than that in patients with distal DVT.

A self-administered questionnaire to measure Chinese smokers' cigarette dependence.

A self-administered questionnaire for screening cigarette dependence was developed based on a set of 6335 Chinese adult smokers (termed the China Cigarette Dependence Test, CCDT). Both a 20-item version (CCDT-20) and a 7-item of the questionnaire (CCDT-7) were developed following 2-round of tests on their construct validity, test-retest reliability and internal consistency, covering seven dimensions (cigarettes per day, tolerance, withdrawal symptoms, craving, loss of control, regularity, and stereotypy). The results showed that the CCDT-20 and CCDT-7 scores were higher in daily smokers than in occasional smokers, and both were associated with self-rated nicotine dependence, exhaled carbon monoxide (CO), saliva cotinine, and DSM-V. The CCDT-20 and CCDT-7 scales were found to be easier to use by smokers in China and provided a more reliable measure of their cigarette dependence.