Diesel exhaust particulate matter induces GC-1 spg cells oxidative stress by KEAP1-NRF2 pathway and inhibition of ATP5α1 S-sulfhydration.

Diesel exhaust particle (DEP) exposure induces a variety of toxicological effects through oxidative stress and inflammation responses. This research investigated the mechanisms underlying DEP-induced GC-1spg cells oxidative stress by examining ROS accumulation, antioxidant defense systems activation, mitochondrial dysfunction, and the Nrf2/Keap1/HO-1 pathway response. Subsequently, we further evaluated the ATP levels, ATP5α synthase activity and ATP5α synthase S-sulfhydrated modification in DEP-exposed GC-1 spg cells. The results showed that DEP exposure significantly inhibited cell proliferation and viability, increased intracellular ROS production, decreased MMP, down-regulated antioxidant capacity, activated the Nrf2/Keap1/HO-1 pathway. However, DEP-induced oxidative stress was partially alleviated by GSH and exogenous H2S. In addition, DEP exposure induced ATP depletion and ATP5α synthase inactivity in GC-1 spg cells, accompanied by ATP5α synthase S-sulfhydrated modification. In conclusion, our research showed that DEP may incapacitate mitochondria through oxidative stress injury, leading to GC-1 spg cells oxidative stress. This process may be associated with the reduction of ATP5α1 S-sulfhydrated modification. It provides a new perspective for the research of the mechanism related to male reproductive toxicity due to air pollution.

F127-SE-tLAP thermosensitive hydrogel alleviates bleomycin-induced skin fibrosis via TGF-ß/Smad pathway.

BACKGROUND: Skin fibrosis affects the normal function of the skin. TGF-ß1 is a key cytokine that affects organ fibrosis. The latency-associated peptide (LAP) is essential for TGF-ß1 activation. We previously constructed and prepared truncated LAP (tLAP), and confirmed that tLAP inhibited liver fibrosis by affecting TGF-ß1. SPACE peptide has both transdermal and transmembrane functions. SPACE promotes the delivery of macromolecules through the stratum corneum into the dermis. This study aimed to alleviate skin fibrosis through the delivery of tLAP by SPACE. METHODS: The SPACE-tLAP (SE-tLAP) recombinant plasmid was constructed. SE-tLAP was purified by nickel affinity chromatography. The effects of SE-tLAP on the proliferation, migration, and expression of fibrosis-related and inflammatory factors were evaluated in TGF-ß1-induced NIH-3T3 cells. F127-SE-tLAP hydrogel was constructed by using F127 as a carrier to load SE-tLAP polypeptide. The degradation, drug release, and biocompatibility of F127-SE-tLAP were evaluated. Bleomycin was used to induce skin fibrosis in mice. HE, Masson, and immunohistochemistry were used to observe the skin histological characteristics. RESULTS: SE-tLAP inhibited the proliferation, migration, and expression of fibrosis-related and inflammatory factors in NIH-3T3 cells. F127-SE-tLAP significantly reduced ECM production, collagen deposition, and fibrotic pathological changes, thereby alleviating skin fibrosis. CONCLUSION: F127-SE-tLAP could increase the transdermal delivery of LAP, reduce the production and deposition of ECM, inhibit the formation of dermal collagen fibers, and alleviate the progression of skin fibrosis. It may provide a new idea for the therapy of skin fibrosis.

Association between triglyceride-glucose index and colorectal polyps: A retrospective cross-sectional study.

BACKGROUND: Colorectal polyps (CPs) are frequently occurring abnormal growths in the colorectum, and are a primary precursor of colorectal cancer (CRC). The triglyceride-glucose (TyG) index is a novel marker that assesses metabolic health and insulin resistance, and has been linked to gastrointestinal cancers. AIM: To investigate the potential association between the TyG index and CPs, as the relation between them has not been documented. METHODS: A total of 2537 persons undergoing a routine health physical examination and colonoscopy at The First People's Hospital of Kunshan, Jiangsu Province, China, between January 2020 and December 2022 were included in this retrospective cross-sectional study. After excluding individuals who did not meet the eligibility criteria, descriptive statistics were used to compare characteristics between patients with and without CPs. Logistic regression analyses were conducted to determine the associations between the TyG index and the prevalence of CPs. The TyG index was calculated using the following formula: Ln [triglyceride (mg/dL) × glucose (mg/dL)/2]. The presence and types of CPs was determined based on data from colonoscopy reports and pathology reports. RESULTS: A nonlinear relation between the TyG index and the prevalence of CPs was identified, and exhibited a curvilinear pattern with a cut-off point of 2.31. A significant association was observed before the turning point, with an odds ratio (95% confidence interval) of 1.70 (1.40, 2.06), P < 0.0001. However, the association between the TyG index and CPs was not significant after the cut-off point, with an odds ratio (95% confidence interval) of 0.57 (0.27, 1.23), P = 0.1521. CONCLUSION: Our study revealed a curvilinear association between the TyG index and CPs in Chinese individuals, suggesting its potential utility in developing colonoscopy screening strategies for preventing CRC.

Protein phosphorylation and kinases: Potential therapeutic targets in necroptosis.

Necroptosis is a pivotal contributor to the pathogenesis of various human diseases, including those affecting the nervous system, cardiovascular system, pulmonary system, and kidneys. Extensive investigations have elucidated the mechanisms and physiological ramifications of necroptosis. Among these, protein phosphorylation emerges as a paramount regulatory process, facilitating the activation or inhibition of specific proteins through the addition of phosphate groups to their corresponding amino acid residues. Currently, the targeting of kinases has gained recognition as a firmly established and efficacious therapeutic approach for diverse diseases, notably cancer. In this comprehensive review, we elucidate the intricate role of phosphorylation in governing key molecular players in the necroptotic pathway. Moreover, we provide an in-depth analysis of recent advancements in the development of kinase inhibitors aimed at modulating necroptosis. Lastly, we deliberate on the prospects and challenges associated with the utilization of kinase inhibitors to modulate necroptotic processes.

Rapid quantitative evaluation of total polar materials (TPM) in frying oil based on an "off-on" fluorescence viscosity response probe.

The content of total polar material (TPM) is considered as a comprehensive indicator to evaluate the quality of edible oils which should be discarded and no longer be used when TPM content exceeding 27 %. Nevertheless, there is currently a lack of a convenient and efficient TPM detection method, which is a meaningful challenge. With the increase of TPM content, the viscosity of frying oil grows, and the two maintain a satisfactory positive correlation. Consequently, an "off-on" fluorescence probe TCF-PR method based on viscosity-response has been developed. There exists a good linear relationship between the fluorescence intensity of the probe and the TPM content of soybean oil ((R2 = 0.9936) and salad oil (R2 = 0.9878), accompanying with the advantage of fast response (3 s), which means the rapid detection of TPM can be realized to determine the quality of frying oil in the field of food safety.

Regulation of the migration of colorectal cancer stem cells via the TLR4/MyD88 signaling pathway by the novel surface marker CD14 following LPS stimulation.

Cell surface markers are most widely used in the study of cancer stem cells (CSCs). However, cell surface markers that are safely and stably expressed in CSCs have yet to be identified. Colonic CSCs express leukocyte CD14. CD14 binding to the ligand lipopolysaccharide (LPS) is involved in the inflammatory response via the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88) signaling pathway. TLR4 and MyD88 have been reported to promote the proliferation, metastasis and tumorigenicity of colon cancer cells, which is consistent with the characteristics of CSCs. In the present study, the proposed experimental method to detect cell proliferation, metastasis and tumorigenesis was used to confirm that, under LPS stimulation, CD14 promoted the proliferation, migration and tumorigenesis of colonic CSCs via the TLR4/MyD88 signaling pathway. Cell Counting Kit-8 and 5-ethynyl-2'-deoxyuridine assays were used to assess the proliferation and migration of the cells. Colony formation and nude mouse xenograft assays were used to assess the capacity of cells to form tumors. Using western blotting and reverse transcription-quantitative PCR, the mRNA and protein levels of CD14, TLR4 and MyD88 were examined. It was confirmed that CD14 promoted the proliferation, metastasis and tumorigenesis of colon CSCs in response to LPS stimulation via the TLR4/MyD88 signaling pathway, and CD14+ colon cancer cells were successfully isolated and sorted. According to the results of proliferation assay, it was determined that CD14 regulated the LPS-induced proliferation of colon CSCs. CD14, TLR4 and MyD88 protein and mRNA expression was upregulated in colon CSCs in response to LPS stimulation. This indicates a potential novel target for colon CSC-related studies.

Comparison of transfusion rates between robotic- and video-assisted lobectomy: a propensity score matching analysis.

The objective is to compare the perioperative blood transfusion rate and postoperative complications between robot-assisted surgery and thoracoscopic surgery in lung cancer patients. This is a single-center retrospective study. Patients underwent lung cancer minimally invasive resection at Fujian Cancer Hospital from April 1, 2022, to April 30, 2023, were enrolled in this study. Patients were divided into robotic-assisted lobectomy (RAL) and video-assisted lobectomy (VAL) groups according to the surgical methods. Data, including demographics, clinic variables, and endpoint outcomes were collected from the electronic medical record. Propensity score matching (PSM) was performed to analyze the baseline data of patients. The RAL group and the VAL group were matched 1:1. Then, the blood transfusion rates and short-term outcomes of the two groups were compared. A logistic regression was performed to analyze the independent risk factors of perioperative blood transfusion. A total of 558 patients were enrolled in this study. 166 of 558 patients were divided into the RAL group, and 392 patients were into the VAL group. A total of 118 patients were selected and analyzed following propensity score matching. After PSM, there was no difference in perioperative transfusion rates, including RBC transfusion and frozen plasma transfusion, between the VAL and RAL groups (P > 0.05). The RAL group had fewer days of drainage tubes (P = 0.036). There was no difference in other short-term outcomes, including the volume of thoracic drainage, the volume of intraoperative blood loss, the length of hospitalization, and the rate of postoperative pulmonary infection (P > 0.05). Volume of intraoperative blood loss, volume of thoracic drainage, and preoperative hemoglobin were independent risk factors of perioperative red blood cell or frozen plasma transfusion; however, RAL or VAL was not. The study showed that the rates of perioperative blood transfusion were comparable between RAL and VAL. RAL is superior for patient recovery in terms of short-term outcomes.

Association of nonalcoholic fatty liver disease with colorectal adenomatous polyps and non-adenomatous polyps: a cross-sectional study.

AIM: This study aimed to investigate the association between non-alcoholic fatty liver disease (NAFLD) and both colorectal adenomatous polyps and non-adenomatous polyps, in order to provide evidence for the prevention of colorectal cancer (CRC) in patients with NAFLD. METHODS: A retrospective, cross-sectional study was conducted at the First People's Hospital of Kunshan, Jiangsu, China. The study included 3028 adults who underwent abdominal ultrasonography and colonoscopy over a 5 year period. We compared characteristics among patients with adenomatous polyps, non-adenomatous polyps, and without colorectal polyps using descriptive statistics. Logistic regression analyses were used to detect associations between NAFLD with the prevalence of adenomatous polyps and non-adenomatous polyps. NAFLD was determined by abdominal ultrasound. Colorectal polyps were assessed by data in the colonoscopy report and pathology report. RESULTS: A total of 65% of patients with NAFLD had colorectal polys (52% adenomatous polyps and 13% non-adenomatous polyps), and 40% of patients without NAFLD had polyps (29% adenomatous polyps and 11% non-adenomatous polyps). After adjusting for confounding variables, NAFLD was significantly associated with the prevalence of adenomatous in males and females [odds ratio (OR) = 1.8, 95% confidence interval (CI): 1.6-2.2, P  < 0.01], but was not associated with non-adenomatous polyps (OR = 1.2, 95% CI:0.9-1.5, P  > 0.05). CONCLUSION: NAFLD is associated with an increased risk of colorectal adenomatous polyps compared to the absence of polyps, but not associated with an increased risk of non-adenomatous polyps. These results provide important evidence for the prevention of CRC in patients with NAFLD.

Brefeldin A from the Deep-Sea-Derived Fungus Fusarium sp. Targets on RIPK3 to Inhibit TNFα-Induced Necroptosis.

From the deep-sea-derived Fusarium sp. ZEN-48, four known compounds were obtained. Their structures were established by extensive analyses of the NMR, HR-ESI-MS, and the X-ray crystallographic data as brefeldin A (BFA, 1), brevianamide F (2), N-acetyltryptamine (3), and (+)-diaporthin (4). Although BFA was extensively investigated for its potent bioactivities, its role on TNFα-induced necroptosis was incompletely understood. In this study, BFA showed significant inhibition on TNFα-induced necroptosis by disrupting the necrosome formation and suppressing the phosphorylation of RIPK3 and MLKL (IC50 =0.5 µM). While, it had no effect on TNFα-induced NF-κB/MAPKs activation and apoptosis. The finding raised significant implications of BFA for necroptosis-related inflammatory disease therapy and new drug development from marine fungi.

Recombinant truncated latency-associated peptide alleviates liver fibrosis in vitro and in vivo via inhibition of TGF-ß/Smad pathway.

BACKGROUND: Liver fibrosis is a progressive liver injury response. Transforming growth factor ß1 (TGF-ß1) is oversecreted during liver fibrosis and promotes the development of liver fibrosis. Therapeutic approaches targeting TGF-ß1 and its downstream pathways are essential to inhibit liver fibrosis. The N-terminal latency-associated peptide (LAP) blocks the binding of TGF-ß1 to its receptor. Removal of LAP is critical for the activation of TGF-ß1. Therefore, inhibition of TGF-ß1 and its downstream pathways by LAP may be a potential approach to affect liver fibrosis. METHODS: Truncated LAP (tLAP) plasmids were constructed. Recombinant proteins were purified by Ni affinity chromatography. The effects of LAP and tLAP on liver fibrosis were investigated in TGF-ß1-induced HSC-T6 cells, AML12 cells and CCl4-induced liver fibrosis mice by real time cellular analysis (RTCA), western blot, real-time quantitative PCR (RT-qPCR), immunofluorescence and pathological staining. RESULTS: LAP and tLAP could inhibit TGF-ß1-induced AML12 cells inflammation, apoptosis and EMT, and could inhibit TGF-ß1-induced HSC-T6 cells proliferation and fibrosis. LAP and tLAP could attenuate the pathological changes of liver fibrosis and inhibit the expression of fibrosis-related proteins and mRNAs in CCl4-induced liver fibrosis mice. CONCLUSION: LAP and tLAP could alleviate liver fibrosis in vitro and in vivo via inhibition of TGF-ß/Smad pathway. TLAP has higher expression level and more effective anti-fibrosis activity compared to LAP. This study may provide new ideas for the treatment of liver fibrosis.

Transepithelial Transport of YWDHNNPQIR and Its Metabolic Fate with Cytoprotection against Oxidative Stress in Human Intestinal Caco-2 Cells.

Studies on antioxidant peptides extracted from foodstuff sources have included not only experiments to elucidate their chemical characteristics but also to investigate their bioavailability and intracellular mechanisms. This study was designed to clarify the absorption and antioxidative activity of YWDHNNPQIR (named RAP), which is derived from rapeseed protein using a Caco-2 cell transwell model. Results showed that 0.8% RAP (C0 = 0.2 mM, t = 90 min) could maintain the original structure across the Caco-2 cell monolayers via the intracellular transcytosis pathway, and the apparent drug absorption rate (Papp) was (6.6 ± 1.24) × 10-7 cm/s. Three main fragments (WDHNNPQIR, DHNNPQIR, and YWDHNNPQ) and five modified peptides derived from RAP were found in both the apical and basolateral side of the Caco-2 cell transwell model. Among these new metabolites, WDHNNPQIR had the greatest antioxidative activity in Caco-2 cells apart from the DPPH assay. With a RAP concentration of 200 µM, there were significant differences in four antioxidative indicators (T-AOC, GSH-Px, SOD, and MDA) compared to the oxidative stress control (P < 0.05). In addition, RAP may also influence apoptosis of the Caco-2 cells, which was caused by AAPH-induced oxidative damage.

Separation and purification of an anti-tumor peptide from rapeseed (Brassica campestris L.) and the effect on cell apoptosis.

Rapeseed peptides were prepared by means of the combined methods of the laboratory bacteria enzyme synergy and the solid-state fermentation of rapeseed meal. The rapeseed peptides were separated and purified with the tumor cell in vitro anti-proliferative activity as an index. Moreover, a kind of rapeseed peptide component RSP-4-3-3 (rapeseed anti-tumor peptide RSP-4-3-3) with high activity was selected. Furthermore, by using reversed-phase high performance liquid chromatography (RP-HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS/MS), the analysis result of its possible amino acid sequence showed that it was Trp-Thr-Pro (408.2 Da). Inverted microscope observation technology and western blot experiments were applied to explore the antitumor impact of the rapeseed peptide RSP-4-3-3 on tumor cells. The results showed that the rapeseed antitumor peptide RSP-4-3-3 could significantly change the morphological features of the HepG2 cells in vitro and cause apoptosis, thus inhibiting the proliferation of the HepG2 cells.

Toxicology and safety of antioxidant of bamboo leaves. Part 2: developmental toxicity test in rats with antioxidant of bamboo leaves.

The antioxidant of bamboo leaves (AOB) is a kind of polyphenols-rich extract from bamboo leaves of the Phyllostachys Sieb. et Zucc. family. It has been certificated as a natural antioxidant by the Ministry of Health of the People's Republic of China in 2003, which has a warrant for use in edible oil, meat product, aquatic product and puffed food as a novel food additive. For safely using AOB, it was required to systemic safety evaluation studies. As part of an extensive program of safety evaluation studies, the traditional teratogenicity test and the reproduction study were conducted in Sprague-Dawley rats. In the traditional teratogenicity test, the treatment was well tolerated and no mortality occurred at doses of 0, 1.43, 2.87 and 4.30 g/kg bw per day; weight gain during gestation, food consumption, and food efficiency were similar in all groups; reproductive performance was not effected by the treatment; examination of the fetuses for external, visceral, and skeletal alterations did not reveal any fetotoxic, embryotoxic, or teratogenic effects of AOB. Therefore, a nominal dietary AOB level of 4.30 g/kg bw per day (The value of intended use and potential exposure to human is 860 mg/60 kg bw per day dependent on the period of the study.) was considered to be the no-observed-adverse-effect level following daily oral administration of AOB. The results of our studies indicate safety of AOB and support the use of AOB as a food additive.