Mitochondrial creatine kinase 1 regulates the cell cycle in non-small cell lung cancer via activation of cyclin-dependent kinase 4.

BACKGROUND: Non-small cell lung cancer (NSCLC) is the main type of the most common malignant tumor in the world. Previous studies have shown that the expression level of mitochondrial creatine kinase 1 (CKMT1) is abnormal in NSCLC, but the mechanism of its effect remains unclear. Therefore, in this study, we intend to clarify the potential mechanism of CKMT1 in NSCLC and provide the theoretical basis for the clinical application of CKMT1. METHODS: The function of CKMT1 in NSCLC was identified by analyzing the GEO dataset and evaluating using in vitro and in vivo models. Protein mass spectrometry was used to find proteins interacting with CKMT1, and Co-immunoprecipitation (Co-IP) and GST-pull down experiments were used to verify the interaction between proteins. The immunofluorescence (IF) assay was used to explore the functional position of CKMT1 in cells. The effect of CKMT1 expression level on the efficacy of paclitaxel (TAX) in the treatment of NSCLC was analyzed by a combined TAX experiment in vivo and in vitro. RESULTS: CKMT1 expression was increased in NSCLC and CKMT1 promoted the proliferation of NSCLC cells in vitro and in vivo. CKMT1 knockdown resulted in a significantly increased G0/G1 fraction and decreased S phase cell fraction, indicating G1 phase arrest. Mechanically, the cyclin-dependent kinase 4 (CDK4) was identified to interact with CKMT1, and the crucial binding areas were focused on the DH domain of CKMT1 and the N- and C-terminal of CDK4. A fraction of the CDK4 proteins colocalize and interact with the CKMT1 at mitochondria, the level of phosphorylated CDK4 was regulated by CKMT1. Hence, the decrease in CKMT1 expression level could increase the antitumor effect of G2/M cell cycle antagonist-TAX in NSCLC in vitro and in vivo. CONCLUSIONS: CKMT1 could interact with CDK4 in mitochondria and regulate the phosphorylated level of CDK4, thus contributing to the proliferation and cell cycle transition of NSCLC cells. And CKMT1 could be a potential target to improve the sensitivity of chemotherapy based on TAX.

Encephaloduroarteriosynangiosis (EDAS) treatment of moyamoya syndrome: evaluation by computed tomography perfusion imaging.

OBJECTIVE: To explore the value of computed tomography perfusion (CTP) imaging for evaluating the efficacy of encephaloduroarteriosynangiosis (EDAS) treatment of moyamoya syndrome (MMS). METHODS: Forty-three patients with MMS (48 hemispheres) who received EDAS treatment were examined using CTP and DSA before and after surgery. CTP of the ipsilateral cortex, contralateral mirror area, and pons region were measured, and the relative cerebral blood flow (rCBF) and volume (rCBV), mean transit time (rMTT), and time-to-peak (rTTP) were calculated. Based on postoperative DSA, 48 hemispheres were apportioned to two groups based on rich (grades 2, 3) or poor (grades 0, 1) collateral vessel formation, and the pre- and post-operative differences in perfusion changes were compared. The association between clinical outcome, CTP, and the degree of DSA collateral vessels was explored. RESULTS: rCBF and rMTT significantly improved in both the poor and rich collateral vessel formation groups (n = 21 and 27, respectively), while rTTP significantly improved only in the latter. Postoperative CTP improved in the rich and the grade 1 collateral vessel groups (p < 0.01). The clinical improvement was consistent with the improvement of CTP (p = 0.07), but less consistent with the degree of collateral angiogenesis (p = 0.003). CONCLUSION: CTP can quantitatively evaluate the improvement of brain tissue perfusion in the operated area after EDAS. Brain tissue perfusion in operated areas improved regardless of postoperative rich or poor collateral vessel formation observed via DSA. A significant improvement in rTTP in the operated area may indicate the formation of abundant collateral vessels. KEY POINTS: • CTP showed that brain tissue perfusion in the operated area after EDAS improved regardless of rich or poor collateral vessel formation observed via DSA. • Significant improvement of rTTP in the operated area may indicate the formation of abundant collateral vessels.

Level, source, and risk assessment of toxic elements in traditional agricultural soils and coping strategies.

The level, source, and risk of toxic elements in traditional agricultural soils are particularly crucial for the sustainable development of agriculture. An important agricultural production base was selected, a total of 251 topsoil samples were collected, eight toxic elements (As, Cd, Cr, Cu, Hg, Ni, Pb, Zn) in soil were analyzed, and environmental and health risk assessments were conducted. Results showed that all concentrations of eight elements in soil samples were lower than the risk screening values with negligible pollution risk. Approximately 83.8% of Hg in soil was originated from atmospheric deposition related to industrial emissions, 53.2% of Cd was derived from direct industrial activities, and the other elements came from soil parent materials or agricultural activities. Accumulation risk of As in agricultural products, potential ecological risk from Cd, and As's ingestion risk and Cr's dermal contact risk should be paid more attention. More stricter monitoring and coping countermeasures and strategies should be established to ensure the sustainable development of agriculture.

Analysis of heavy metal content in edible honeysuckle (Lonicera japonica Thunb.) from China and health risk assessment.

Honeysuckle is a commonly used medicine for health care and treatment. To detect heavy metal pollution in honeysuckle from China and quantify the health risk of heavy metal via dietary intake, the Pb, Cd, Cr, As, Hg, Ni, Mn, Cu, and Zn contents in honeysuckle samples were determined by ICP-MS. The dissolution rate of heavy metals in honeysuckle was measured by decoction and soaking. The hazard quotient (HQ) and total hazard index (HI) were used to evaluate the noncarcinogenic risk of nine heavy metals in honeysuckle, and the carcinogenic risks of Cd and As were evaluated using the carcinogen risk. Cd exhibited the maximum permissive limit standard-exceeding rate (40.2%) in honeysuckle, followed by Cu (37.6%) and Pb (8.5%). As and Hg did not exceed the standard values, and Cr, Ni, Mn, and Zn had no limits. In a decoction fluid after 30 min of boiling, the transfer rates of Pb, Cd, As, Ni, Mn, Cu, and Zn ranged from 11.9% to 19.9%, whereas that of Cr was low (1.0%). In a soaking fluid, the transfer rates ranged from 17.0% to 56.9%; no transfer rate was detected for Hg in neither the decoction fluid nor the soaking fluid. In addition, the 95th percentile Rs of As and Cd in honeysuckle were 5.93 × 10-6 and 8.12 × 10-5, respectively. The carcinogenic risk of Cd at 56.99th percentile reached the threshold set by the World Health Organization (1.0 × 10-5). The results showed that intake of Pb, Cd, Cr, As, Hg, Ni, Mn, Cu, and Zn by the human body through honeysuckle could not cause noncarcinogenic damage. The element As had no carcinogenic risk, but Cd had a carcinogenic risk to a certain extent.

Fast determination of 14 mycotoxins in chestnut by dispersive solid-phase extraction coupled with ultra high performance liquid chromatography-tandem mass spectrometry.

A dispersive solid-phase extraction coupled with ultra high performance liquid chromatography with tandem mass spectrometry method was developed and validated for the simultaneous determination of T-2 toxin, penicillic acid, fumonisins B1 , B2 , and B3 , aflatoxins B1 , B2 , G1 , and G2 , ochratoxin A, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, and zearalenone in chestnut samples. The method was used to analyze 136 samples obtained from Shandong province in China. The mycotoxins were extracted using a dispersive solid-phase extraction method and cleaned using an improved quick, easy, cheap, effective, rugged, and safe approach. The mycotoxins were then detected using a triple-quadrupole mass spectrometer. The limits of detection and quantification ranged from 0.02 to 1 and 0.1 to 2 µg/kg, respectively. The recovery rates ranged from 74.2 to 109.5%, with relative standard deviations below 15%. A total of 71 samples were contaminated with seven mycotoxins at concentrations ranging from 1.2 to 105.5 µg/kg, with a number of samples exceeding the maximum limits set in the European regulations for mycotoxins in unprocessed chestnuts.

Significance of nuclear magnetic resonance combined with Ki-67 and VEGF detection in the diagnosis and prognosis evaluation of brain glioma.

PURPOSE: To investigate the significance of nuclear magnetic resonance (NMR) combined with Ki-67 and vascular endothelial growth factor (VEGF) detection in the diagnosis and prognosis evaluation of brain glioma. METHODS: 78 patients with brain glioma treated at the Affiliated Hospital of Jining Medical University from January 2015 to August 2017 were studied. All patients underwent the NMR diffusion tensor imaging examination. The expressions of Ki-67 and VEGF in brain glioma tissues were detected using immunohistochemistry and survival analyses were performed for patients in high apparent diffusion coefficient (ADC) group and low ADC group, high-expression Ki-67 and VEGF group and low-expression Ki-67 and VEGF group before treatment. Moreover, the value of combined diagnosis in the prognosis evaluation of patients was analyzed. RESULTS: NMR diffusion tensor imaging showed that the fractional anisotropy (FA) value in the tumor enhancement region on the affected side was significantly lower than in the contralateral normal region, but ADC was significantly higher in the contralateral normal region; the FA value in grade I-II brain glioma enhancement region was higher compared with grade III-IV glioma enhancement region, but ADC was lower in the grade-III-IV glioma enhancement region (p<0.05). The low expression rates of Ki-67 and VEGF in patients with grade I-II brain glioma were significantly higher than in patients with grade III-IV glioma (p<0.05). After treatment, the 3-year survival rate of high ADC group was lower than that of low ADC group, and the 3-year survival rate of high-expression Ki-67 and VEGF group was also obviously lower than that in low-expression Ki-67 and VEGF group (p<0.05). Besides, the area under the receiver operating characteristic (ROC) curve of NMR combined with Ki-67 and VEGF detection in the prediction of patient prognosis was 0.906, the sensitivity 91.6%, and the specificity 89.5%. CONCLUSION: NMR diffusion tensor imaging has a high application value in the diagnosis of brain glioma. The expressions of Ki-67 and VEGF are related to the pathological grade of glioma, which can be used as biological indexes for the diagnosis of glioma. Moreover, the combined detection of the three items can not only accurately determine the grade of glioma malignancy, but also effectively evaluate the prognosis of patients, thus providing a scientific basis for the selection of therapeutic regimen.

Evaluation of surgical revascularization procedure outcomes for adult Moyamoya disease: a computed tomography perfusion-based study.

BACKGROUND: The effectiveness of surgical interventions, whether direct or indirect, for Moyamoya disease (MMD) remains controversial. This study aims to investigate CT perfusion (CTP) as an objective method to evaluate the outcomes of different surgical modalities for adult MMD. METHODS: The clinical and imaging data of 41 patients who underwent superficial temporal artery-middle cerebral artery (STA-MCA) bypass and 43 who received encephaloduroarteriosynangiosis (EDAS) were retrospectively analyzed. Intra- and intergroup differences in the Modified Rankin Scale (mRS) score, the change in clinical symptoms, collateral grade, and CTP parameters pre- and postoperatively were compared. RESULTS: The overall level of the change in clinical symptoms in the STA-MCA group was higher than in the EDAS group (p < 0.05). In the operative area, the relative cerebral blood flow (rCBF) was significantly higher whereas the relative time to peak (rTTP) and the relative mean transit time (rMTT) were significantly lower in the STA-MCA and EDAS groups postoperatively than preoperatively (all p < 0.05). In the ipsilateral frontal lobe and basal ganglia, the postoperative rCBF was significantly higher, and the rTTP was significantly lower than the preoperative in the STA-MCA group (all p < 0.05). The postoperative rCBF improvement was higher in each brain area for STA-MCA than in the EDAS group (all p < 0.05). CONCLUSION: Highlighting the utility of CTP, this study demonstrates its effectiveness in assessing postoperative cerebral hemodynamic changes in adult MMD patients. STA-MCA yielded a larger postoperative perfusion area and greater improvement compared to EDAS, suggesting CTP's potential to elucidate symptom variation between two surgical revascularization procedures. CRITICAL RELEVANCE STATEMENT: We analyzed computed tomography perfusion parameters in pre- and postoperative adult Moyamoya disease patients undergoing superficial temporal artery-middle cerebral artery bypass and encephaloduroarteriosynangiosis. Our findings suggest computed tomography perfusion's potential in objectively elucidating symptom variations between these surgical revascularization approaches for MMD. KEY POINTS: • Postoperative perfusion improvement is only confined to the operative area after EDAS. • Besides the operative area, postoperative perfusion in the ipsilateral frontal lobe and basal ganglia was also improved after STA-MCA. • The degree of perfusion improvement in each brain area in the STA-MCA group was generally greater than that in the EDAS group.

Disulfiram enhances cisplatin cytotoxicity by forming a novel platinum chelate Pt(DDTC)3.

Despite the use of targeted therapy in non-small cell lung cancer (NSCLC) patients, cisplatin (DDP)-based chemotherapy is still the main option. However, DDP resistance is the major factor contributing to the failure of chemotherapy. In this study, we tried to screen DDP sensitizers from an FDA-approved drug library containing 1374 small-molecule drugs to overcome DDP resistance in NSCLC. As a result, disulfiram (DSF) was identified as a DDP sensitizer: DSF and DDP had synergistic anti-NSCLC effects, which are mainly reflected in inhibiting tumor cell proliferation, plate colony formation and 3D spheroidogenesis and inducing apoptosis in vitro, as well as the growth of NSCLC xenografts in mice. Although DSF has recently been reported to promote the antitumor effect of DDP by inhibiting ALDH activity or modulating some important factors or pathways, unexpectedly, we found that DSF reacted with DDP to form a new platinum chelate, Pt(DDTC)3+, which might be one of the important mechanisms for their synergistic effect. Moreover, Pt(DDTC)3+ has a stronger anti-NSCLC effect than DDP, and its antitumor activity is broad-spectrum. These findings reveal a novel mechanism underlying the synergistic antitumor effect of DDP and DSF, and provide a drug candidate or a lead compound for the development of a new antitumor drug.

Risk Assessment of Chlorothalonil as a Probable Human Carcinogen on Selected Vegetables in an Eastern China Province.

Objectives: This study aimed to provide an assessment of chlorothalonil's possible carcinogenic risk posed to the public. In combination and comparison with the non-carcinogenic risk, the results hopefully could provide useful insights, early warning, and references for policy formulation. Methods: This study firstly investigated the occurrence of chlorothalonil on selected key vegetables for different scenarios, and then conducted an exposure assessment with officially published data. Lastly, both the non-carcinogenic and carcinogenic risk of chlorothalonil were calculated by using Monte-Carlo simulation. Results: Even though mean non-carcinogenic risks of chlorothalonil for all scenarios were below threshold value, the mean carcinogenic risks for maximum-risk scenario and most-likely risk scenario were mostly above threshold value. High probabilities of exceedance of threshold value existed for carcinogenic risk under all scenarios. Conclusion: Potential threat to public health existed for conventionally 'safe' pesticide if considering the possible carcinogenicity. Extra caution should be taken and the potential carcinogenic effects should be included into consideration for better protection of public health during the policy formulation process.

Profile of Bacterial Community and Antibiotic Resistance Genes in Typical Vegetable Greenhouse Soil.

The use of vegetable greenhouse production systems has increased rapidly because of the increasing demand for food materials. The vegetable greenhouse production industry is confronted with serious environmental problems, due to their high agrochemical inputs and intensive utilization. Besides this, antibiotic-resistant bacteria, carrying antibiotic-resistance genes (ARGs), may enter into a vegetable greenhouse with the application of animal manure. Bacterial communities and ARGs were investigated in two typical vegetable-greenhouse-using counties with long histories of vegetable cultivation. The results showed that Proteobacteria, Firmicutes, Acidobacteria, Chloroflexi, and Gemmatimonadetes were the dominant phyla, while aadA, tetL, sul1, and sul2 were the most common ARGs in greenhouse vegetable soil. Heatmap and principal coordinate analysis (PCoA) demonstrated that the differences between two counties were more significant than those among soils with different cultivation histories in the same county, suggesting that more effects on bacterial communities and ARGs were caused by soil type and manure type than by the accumulation of cultivation years. The positive correlation between the abundance of the intI gene with specific ARGs highlights the horizontal transfer potential of these ARGs. A total of 11 phyla were identified as the potential hosts of specific ARGs. Based on redundancy analysis (RDA), Ni and pH were the most potent factors determining the bacterial communities, and Cr was the top factor affecting the relative abundance of the ARGs. These results might be helpful in drawing more attention to the risk of manure recycling in the vegetable greenhouse, and further developing a strategy for practical manure application and sustainable production of vegetable greenhouses.

Evaluation of canthaxanthin in eggs and its subsequent dietary risks to Chinese consumers.

Canthaxanthin is a colourant widely used in medicine, food and cosmetics, which is closely related to human eye dysfunction. The EU determines that the daily acceptable intake of canthaxanthin (ADI) is 0.03 mg canthaxanthin per kg body weight. In order to protect people's diet health and avoid the occurrence of food safety accidents, this paper measured the content of canthaxanthin in 114 egg samples sourced from different regions of China using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The dietary exposure assessment method was used to evaluate the dietary risk to Chinese consumers. Results showed that the content of canthaxanthin varies significantly across regions of origin, with eggs produced in Beijing having a lower amount than those from other regions. The average dietary exposure of children (2-10 years old) was 0.242 ~ 0.653 µg/(kg bw∙d), and the dietary exposure of 95/97.5/99 percentile was 0.246 ~ 1.779 µg/(kg bw∙d). The maximum exposure was 3.089 µg/(kg bw∙d), and its risk quotient was 0.103 (<1), which was an acceptable risk. For the teenager group (11-18 years old), the average dietary exposure was 0.141 ~ 0.247 µg/(kg bw∙d), and the dietary exposure at the 95/97.5/99 percentile was 0.143 ~ 0.671 µg/(kg bw∙d), presenting an acceptable risk. For adults, the dietary risk of eating eggs with canthaxanthin is the lowest among the three age groups.

High-Yield Method To Fabricate and Functionalize DNA Nanoparticles from the Products of Rolling Circle Amplification.

DNA condensation is a facile method to construct DNA nanostructure with a high biostability and low cost, which is mainly used in DNA separation and gene transfection. The recent emerging condensed DNA nanostructures from the rolling circle amplification (RCA), i.e., the complexes between RCA products and magnesium pyrophosphate (RCA-MgPPi), have quickly become attractive biomedical materials with broad application potential because they combine the advantages of the designable and high-throughput isothermal amplification technique and the high stability of DNA condensation structures. However, we find that only approximately 10% of RCA products can be condensed after an RCA reaction, which limits the practical application of the RCA-MgPPi nanostructures. Therefore, in this paper, we investigate how to control the condensation efficiency of RCA-synthesized DNAs in depth. The very long RCA products, which show high charge densities, can be efficiently condensed by an excessive amount of Mg2+ to form RCA-MgPPi nanostructures at a yield approaching 100%. Additionally, the new condensation approach is general and is not limited to the RCA products, which can be applied to other polymeric DNAs. These RCA-MgPPi nanoparticles exhibit a high biostability and low toxicity, in addition, which can be efficiently functionalized with foreign components to create hierarchical properties. Finally, as a proof of concept, based on RCA-MgPPi nanostructures, a ratiometric fluorescence sensor system has been constructed and demonstrated to be an efficient lysosomal pH tracker.

Magnesium-Stabilized Multifunctional DNA Nanoparticles for Tumor-Targeted and pH-Responsive Drug Delivery.

Functional nucleic acids, which can target cancer cells and realize stimuli-responsive drug delivery in tumor microenvironment, have been widely applied for anticancer chemotherapy. At present, high cost, unsatisfactory biostability, and complicated fabrication process are the main limits for the development of DNA-based drug-delivery nanocarriers. Here, a doxorubicin (Dox)-delivery nanoparticle for tumor-targeting chemotherapy is developed taking advantage of rolling circle amplification (RCA) technique, by which a high quantity of functional DNAs can be efficiently collected. Furthermore, Mg2+, a major electrolyte in human body showing superior biocompatibility, can sufficiently condense the very long sequence of an RCA product and better preserve its functions. The resultant DNA nanoparticle exhibits a high biostability, making it a safe and ideal nanomaterial for in vivo application. Through cellular and in vivo experiments, we thoroughly demonstrate that this kind of Mg2+-stabilized multifunctional DNA nanoparticles can successfully realize tumor-targeted Dox delivery. Overall, exploiting RCA technique and Mg2+ condensation, this new strategy can fabricate nanoparticles with a nontoxic composition through a simple fabrication process and provides a good way to preserve and promote DNA functions, which will show a broad application potential in the biomedical field.