Phospholipase D Mediates Glutamine-Induced mTORC1 Activation to Promote Porcine Intestinal Epithelial Cell Proliferation.

BACKGROUND: The intestinal epithelium is one of the fastest self-renewal tissues in the body, and glutamine plays a crucial role in providing carbon and nitrogen for biosynthesis. In intestinal homeostasis, phosphorylation-mediated signaling networks that cause altered cell proliferation, differentiation, and metabolic regulation have been observed. However, our understanding of how glutamine affects protein phosphorylation in the intestinal epithelium is limited, and identifying the essential signaling pathways involved in regulating intestinal epithelial cell growth is particularly challenging. OBJECTIVES: This study aimed to identify the essential proteins and signaling pathways involved in glutamine's promotion of porcine intestinal epithelial cell proliferation. METHODS: Phosphoproteomics was applied to describe the protein phosphorylation landscape under glutamine treatment. Kinase-substrate enrichment analysis was subjected to predict kinase activity and validated by qRT-PCR and Western blotting. Cell Counting Kit-8, glutamine rescue experiment, chloroquine treatment, and 5-fluoro-2-indolyl deschlorohalopemide inhibition assay revealed the possible underlying mechanism of glutamine promoting porcine intestinal epithelial cell proliferation. RESULTS: In this study, glutamine starvation was found to significantly suppress the proliferation of intestinal epithelial cells and change phosphoproteomic profiles with 575 downregulated sites and 321 upregulated sites. Interestingly, phosphorylation of eukaryotic initiation factor 4E-binding protein 1 at position Threonine70 was decreased, which is a crucial downstream of the mechanistic target of rapamycin complex 1 (mTORC1) pathway. Further studies showed that glutamine supplementation rescued cell proliferation and mTORC1 activity, dependent on lysosomal function and phospholipase D activation. CONCLUSION: In conclusion, glutamine activates mTORC1 signaling dependent on phospholipase D and a functional lysosome to promote intestinal epithelial cell proliferation. This discovery provides new insight into regulating the homeostasis of the intestinal epithelium, particularly in pig production.

Piceatannol Alleviates Deoxynivalenol-Induced Damage in Intestinal Epithelial Cells via Inhibition of the NF-κB Pathway.

Deoxynivalenol (DON) is a common mycotoxin that is widely found in various foods and feeds, posing a potential threat to human and animal health. This study aimed to investigate the protective effect of the natural polyphenol piceatannol (PIC) against DON-induced damage in porcine intestinal epithelial cells (IPEC-J2 cells) and the underlying mechanism. The results showed that PIC promotes IPEC-J2 cell proliferation in a dose-dependent manner. Moreover, it not only significantly relieved DON-induced decreases in cell viability and proliferation but also reduced intracellular reactive oxygen species (ROS) production. Further studies demonstrated that PIC alleviated DON-induced oxidative stress damage by increasing the protein expression levels of the antioxidant factors NAD(P)H quinone oxidoreductase-1 (NQO1) and glutamate-cysteine ligase modifier subunit (GCLM), and the mRNA expression of catalase (CAT), Superoxide Dismutase 1 (SOD1), peroxiredoxin 3 (PRX3), and glutathione S-transferase alpha 4 (GSTα4). In addition, PIC inhibited the activation of the nuclear factor-B (NF-κB) pathway, downregulated the mRNA expression of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) to attenuate DON-induced inflammatory responses, and further mitigated DON-induced cellular intestinal barrier injury by regulating the protein expression of Occludin. These findings indicated that PIC had a significant protective effect against DON-induced damage. This study provides more understanding to support PIC as a feed additive for pig production.

E17241 as a Novel ABCA1 (ATP-Binding Cassette Transporter A1) Upregulator Ameliorates Atherosclerosis in Mice.

[Figure: see text].

[Expression and regulatory role of ultraconserved long non-coding RNA uc.77 in lung cancer].

Objective: To investigate the effect and molecular mechanism of ultra-conservative long non-coding RNA uc.77 in lung cancer. Methods: Lung cancer tissues and adjacent normal tissues were obtained from 61 patients with lung cancer who were diagnosed with lung cancer and underwent surgery from 2014 to 2016 in the General Hospital of the Southern Theater Command of the People's Liberation Army. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the uc.77 relative expressions in normal human bronchial epithelial cells 16HBE, lung cancer cell lines, and 61 pair lung cancer tissues. Uc.77 siRNA was transfected into lung cancer cells to interfere with the expression of uc.77, qRT-PCR was used to verify the interference effect, CCK8 method and clone formation experiment were used to detect cell proliferation ability, flow cytometry was used to detect apoptosis and cell cycle changes. H1299 cells transfected with uc.77 siRNA were injected into the subcutaneous right side of BALB/c nude mice to construct a tumor-bearing model for exploring the role of uc.77 on tumor growth. Western blot and qRT-PCR methods were used to detect the protein and mRNA expressions of p21. Results: The relative expression levels of uc.77 in lung cancer cell lines 95D, H1299, A549, H460, H446 and 16HBE-T were significantly higher than that of 16HBE cells (P<0.05). The uc.77 RNA expression levels of lung cancer tissues was significantly higher than that of the adjacent normal tissues (P<0.001). In addition, increased lncRNA uc.77 expression was significantly associated with big tumor size, lymph node metastasis and advanced TNM stage (P<0.05). After transfection with uc.77 siRNA, the expressions of uc.77 in H1299, 95-D and 16HBE-T cells were reduced (P<0.05), and the cell proliferation capacities were reduced at 48 hours and 72 hours (P<0.05). After transfection with uc.77 siRNA-1, the G(0)/G(1) phase cell ratio of H1299 siRNA-1 group [(71.86±3.46)%] was higher than those of H1299-control group [(47.62±5.48)%] and H1299 siRNA-NC group [(61.38±5.62)%, P<0.05], S phase cell ratio of H1299 siRNA-1 group [(14.99±3.61)%] was lower than those of H1299-control group [(34.95±7.05)%] and H1299 siRNA-NC group [(23.75±5.87)%, P<0.05], the apoptosis rate of H1299 siRNA-1 group [(4.90±1.80)%] was higher than those of H1299-control group [(3.30±0.80)%] and H1299 siRNA-NC group [(2.80±1.20)%, P<0.05], the colony formation rate of H1299 siRNA-1 group [(19.20±2.00)%] was lower than those of H1299 control group [(32.60±2.00)%] and H1299 siRNA-NC group [(34.40±1.00)%, P<0.05]. The results of the nude mice tumor formation experiment showed that the tumor volume of the H1299 siRNA-1 group was significantly lower than those of the H1299-control group and the H1299-negative control group (P<0.05), the average tumor weight of H1299 siRNA-1 group was significantly lower than those of H1299-control group and H1299-negative control group (P<0.05), tumor cell growth marker Ki-67 in the H1299 siRNA-1 group showed weak positive, and Ki-67 in the H1299-control group and H1299-negative control group showed positive. The result of qRT-PCR analysis showed that the mRNA expression level of p21 in H1299 siRNA-1 group (2.57±0.45) was higher than those in H1299 control group (1.00±0.00, P=0.001) and H1299 siRNA-NC group (1.52±0.37, P=0.009). The result of western blotting analysis also showed that the expression of p21 protein level in H1299 siRNA-1 group increased. Conclusions: The expression of ultraconserved long non-coding RNA uc.77 is elevated in lung cancer cell lines and lung cancer tissues. Silencing the expression of ultraconservative long noncoding RNA uc.77 can inhibit tumor growth, and blocking uc.77 expression may be a potential therapeutic target for lung cancer.

[Introduction and implications of WHO position paper: vaccines against influenza, May 2022].

On May 13, 2022, World Health Organization(WHO) Position Paper on Influenza Vaccine (2022 edition) was published. This position paper updates information on influenza epidemiology, high risk population, the impact of immunization on disease, influenza vaccines and effectiveness and safety, and propose WHO's position and recommendation that all countries should consider implementing seasonal influenza vaccine immunization programmes to prepare for an influenza pandemic. In addition, it proposes that the influenza surveillance platform can be integrated with the surveillance of other respiratory viruses, such as SARS-CoV-2 and Respiratory Syncytial Virus. This position paper has some implications for the prevention and control of influenza and other respiratory infectious diseases in China: (1) Optimize influenza vaccine policies to facilitate the implementation of immunization services; (2) Influenza prevention and control should from the perspective of Population Medicine focus on the individual and community to integrate with "Promotion, Prevention, Diagnosis, Control, Treatment, Rehabilitation"; (3) Incorporate prevention and control of other respiratory infectious diseases such as influenza, COVID-19, respiratory syncytial virus and adenovirus, and intelligently monitor by integrating multi-channel data to achieve the goal of co-prevention and control of multiple diseases.

Rapid Determination of Ammonia in Biological Samples by GC-MS Derivatization Method.

ABSTRACT: Objective To establish a qualitative and quantitative method to determine ammonia in biological samples by gas chromatography-mass spectrometry （GC-MS）. Methods A heptafluorobutyryl chloride derivatization method was used. GC-MS was used for determination. The effects of different pH conditions, derivatization temperature, time and different extraction solvents on the test results were investigated. The pretreatment conditions were optimized. Results This method could accurately detect the ammonia content in blood, and the limit of detection was determined to be 0.1 µg/mL. The target component showed good linearity in the range of 0.5-200.0 µg/mL （R2=0.987 7）. The relative standard deviation range of intra-day precision was 2.59%-3.88%. The relative standard deviation range of inter-day precision was 3.21%-3.76%. Conclusion The method showed good sensitivity, stability and specificity, therefore can be used for forensic toxicology analysis and clinical biochemical detection.

[Research Progress on Estimation of Postmortem Submersion Interval].

ABSTRACT: Postmortem interval （PMI） estimation is one of the most important and difficult academic tasks in forensic sciences. Due to the influence of the corpse itself and the water environment, corpses in water have unique corruption phenomenon and laws. Based on the experience of traditional PMI studies of corpses on land, forensic practitioners across the world have proposed a variety of practical methods for estimating postmortem submersion interval （PMSI）. This paper summarizes the literatures related to PMSI in recent years, and introduces methods to infer PMSI according to the phenomenon of corpses, the development of insects, the succession pattern of aquatic organisms, and the changes of other physical and chemical indexes of corpses, in order to provide some reference for the study of PMSI of corpses in water.

Rapid Identification of Four New Synthetic Cannabinoids in Whole Blood.

ABSTRACT: Objective To establish accurate and rapid methods to identify four new synthetic cannabinoids （JWH-203, JWH-122, 5F-APINACA and AB-CHMINACA） in blood samples. Methods The whole blood samples were extracted by acetonitrile and methanol, screened by gas chromatography-mass spectrometry （GC-MS） then confirmed by liquid chromatography-tandem mass spectrometry （LC-MS/MS） method, and multiple reaction monitoring （MRM） mode was used for quantitative analysis. Results The GC-MS method needed 21 min to complete the analysis, while the LC-MS/MS method needed 5 min. The AB-CHMINACA, JWH-203, 5F-APINACA and JWH-122 all used quasi molecular ion peak as a parent ion. The precursor-product ion combinations were m/z 357.4→312.2, m/z 340.2→125.0, m/z 384.1→135.1 and m/z 356.4→169.2. The four synthetic cannabinoids in blood samples had good linearity in the 1-250 ng/mL mass concentration range （r>0.99）. The limits of detection （LODs） were in the range of 0.1-0.5 ng/mL, the recovery rate was 85.4%-95.2%, the RSD less than 10.0%, and the matrix effect was 80.3%-92.8%. Conclusion The GC-MS and LC-MS/MS chromatographic behaviors and mass spectrometry analysis information of four synthetic cannabinoids were obtained in this study, and the possible causes of differences in chromatographic behaviors were discussed preliminarily. Therefore this study has a suggestive effect on judging the development trend of synthetic cannabinoids. This method can be used for rapid identification of four synthetic cannabinoids in blood, which can provide reference for identification of new synthetic cannabinoids when they are proliferating at present.

Decreasing cytokeratin 17 expression in head and neck cancer predicts nodal metastasis and poor prognosis: The first evidence.

OBJECTIVES: Cytokeratins (CKs) are mainly expressed in epithelial carcinomas and are valuable for making diagnoses and identifying metastatic status. Changes in the expression of individual CKs in certain carcinoma may be relevant to establishing a prognosis. However, the prognostic significance of CKs in head and neck squamous cell carcinoma (HNSCC) remains elusive. Herein, we investigated the diverse and unique expression patterns of Cytokeratin 13 (CK13) and Cytokeratin 17 (CK17) and assessed the role of CK17 as a predictor for HNSCC metastasis and prognosis. METHODS: CK13 and CK17 expressions were evaluated using immunohistochemical tissue microarray (TMA) analysis with 106 patients of HNSCC. To clarify the characterisation of CK17 expression with respect to its ability in predicting metastatic disease, an in vitro study of cells migration/invasion assays was conducted. Furthermore, the correlation of CK17 expression to clinicopathologic variables and prognosis was analyzed using a serial statistical method. RESULTS: CK13 was predominately expressed in non-cancerous tissues and was lost in HNSCC. Decreasing expression of CK17 correlated with cancerous cell migration and invasion (P < .0001) in an in vitro study. CK17 expression was lower in the N1 and N2 nodal metastases category compared to the N0 stage. Moreover, Kaplan-Meier survival analyses showed that a lower CK17 expression was associated with a poorer survival connotation in HNSCC patients (P < .05) with 10-year follow-up. CONCLUSION: Our findings provide the first evidence that CK17 under-expression might be a potential predictor of nodal metastasis and adverse prognosis.

Levothyroxine monotherapy versus levothyroxine and selenium combination therapy in chronic lymphocytic thyroiditis.

PURPOSE: New strategies are needed for prevention and treatment of chronic lymphocytic thyroiditis (CLT). This study aimed to assess whether combination of levothyroxine treatment and selenium (Se) supplementation results in improved therapeutic effects in CLT compared with levothyroxine monotherapy. METHODS: An open-label, randomized controlled study was performed in 60 CLT patients assigned to two groups. Levothyroxine group (LT) patients (n = 24) received levothyroxine alone for 3 months; meanwhile, the combination (LTSS) group (n = 36) was administered levothyroxine with selenium yeast capsule. Blood selenium concentrations, anti-thyroid peroxidase (TPO) and anti-thyroglobulin (Tg) antibody levels, and inflammatory cytokine amounts were compared between both groups before and after treatment. RESULTS: At baseline, similar values were obtained in both groups for all the parameters assessed (p > 0.05). After treatment, significantly increased blood selenium levels (µg/L) [90.05 (80.69, 107.76) vs. 39.64 (29.42, 51.10), p < 0.001] and decreased anti-TPO antibody (23.63 ± 9.31 vs. 32.00 ± 10.41%, p = 0.002), anti-Tg antibody (35.84 ± 15.21 vs. 45.47 ± 14.24%, p = 0.015) and IL-2 amounts (pg/mL) [159.29 (124.54, 189.70) vs. 226.48 (190.74, 266.56), p < 0.001] were observed in the LTSS group compared with the LT group post-treatment; meanwhile, similar IL-10 concentrations [23.14 (21.65, 28.56) pg/mL vs. 24.68 (21.71, 29.67) pg/mL] were obtained in both groups. Subgroup analysis of patients with hypothyroidism showed the same trend observed in the whole population; in patients with normal thyroid function, only Se and IL-2 amounts differed between the two treatment groups. Correlation analysis of of the indexes: in HT patients, the basal serum selenium concentration was positively correlated with TT4 (r = 0.294, p < 0.05), significantly negatively correlated with TSH (r = -0.343, p < 0.01), and had no significant correlation with TT3 (p > 0.05). CONCLUSIONS: These findings indicated that levothyroxine and selenium combination results in improved therapeutic effects than the levothyroxine monotherapy in preventing CLT progression.

[Developing and application of an autoverification system for clinical chemistry and immunology test results].

Objective: To develop and validate an autoverification system for biochemistry and immunology test results for application in routine work. Methods: Algorithms was designed and translated into the laboratory information system. Parameters including verify limit, delta check, logic relation between tests was set up in the system. Verification rate of every test and the causes of fails were analyzed, according to which the system and parameters were modified. The autoverified reports were evaluated by chief technicians. Only when all of the autoverified results pass the evaluation, the system applied for routine work of releasing the results. Autoverification rate and turnaround time(TAT) were calculated for evaluation of the efficiency of the system. Results: A brand new autoverification system was developed and applied for routine work. The autoverification rate for each single test was 91.1%-96.6%. The autoverification rate for reports was 74%. With the autoverification system, the media of TAT reduced from 111.6(53.9-270.7) min to 87.2(45.4-202.4) min, whereas the time from instrument finishing analysis to releasing the reports reduced from 18.6(1.0-99.3) min to 0.1(0-58.3)min. The number of staff specified for results validation reduced from three to one. Conclusions: The newly developed system can be used for autoverification of biochemistry and immunology test results. The autoverification system can greatly reduce TAT and raise working efficiency. It's essential to employ carefully designed algorithm, appropriate parameters and comprehensive evaluation when developing a new autoverification system.

[Determination of 1-methylhydantoin Concentration in Blood by GC-MS Method and Its Application in Forensic Medicine].

OBJECTIVES: To establish a gas chromatographic-mass spectrometric （GC-MS） analysis method for quantifying 1-methylhydantoin concentration in whole blood. To provide technical support to forensic identification related cases of 1-methylhydantoin. METHODS: As an internal standard, 500 ng SKF525A was added to 0.5 mL blood sample, and then 2 mL 0.01 mol/L dilute hydrochloric acid and 0.5 g ammonium carbonate were added in order to buffer the pH value to 9, and following 2 mL ethyl acetate. The organic solvent layer was obtained after centrifuge and then analysed by GC-MS after drying. RESULTS: Good linear relationship of 1-methylhydantoin in blood was obtained in the range of 0.5-50 ng/mL. The equation of linear regression was y=0.015 51 x+0.007 26（R²=0.999 7） with 0.1 ng/mL detection limit, and the recovery was 93.02%-108.12%. The intra-day and inter-day precision were less than 6.07% and 13.37%, respectively. CONCLUSIONS: The results gotten by this method is accurate and reproducible, which can be used for the determination of 1-methylhydantoin concentration in blood samples.

Association study between matrix metalloproteinase-9 gene (MMP9) polymorphisms and the risk of Henoch-Schönlein purpura in children.

Henoch-Schönlein purpura nephritis (HSPN), the most serious long-term complication of Henoch-Schönlein purpura, is one of the most common renal diseases in children. Matrix metalloproteinase-9 (MMP-9) is implicated in the pathogenesis of renal diseases. Genomic DNA was isolated from the venous blood leukocytes of 220 unrelated patients with HSPN and 205 unrelated healthy individuals. To identify markers contributing to genetic susceptibility to HSPN, this study examined the potential association between HSPN and four single nucleotide polymorphisms of the MMP-9 gene (MMP9) (rs17576, rs3918254, rs3787268, and rs2236416) by using the MassARRAY system. The allelic or genotypic frequencies of the rs17576 (exon 6) and rs3918254 (intron 6) polymorphisms in HSPN were significantly different from those in the healthy controls. The HSPN subjects had a significantly higher frequency of the G allele of rs17576 (χ(2) = 8.416, P = 0.004, OR = 1.556, 95%CI = 1.153-2.100) and a significantly lower frequency of the A allele of rs2236416 (χ(2) = 10.363, P = 0.001, OR = 0.545, 95%CI = 0.375-0.791). Linkage disequilibrium was observed in two blocks (D' > 0.9; r(2) > 0.8 in control). In block 1, significantly more G-C haplotypes (P = 0.011) and significantly fewer A-C haplotypes (P = 0.008) were found in the HSPN subjects. In block 2, significantly more G-G haplotypes (P = 0.016) and significantly fewer A-G haplotypes (P = 0.006) were found in the HSPN subjects. These observations suggest that the rs17576 and rs3918254 polymorphisms of MMP9 are associated with HSPN.

[Information processing speed and influential factors in multiple sclerosis].

OBJECTIVE: To study the information processing speed and the influential factors in multiple sclerosis (MS) patients. METHODS: A total of 36 patients with relapsing-remitting MS (RRMS), 21 patients with secondary progressive MS (SPMS), and 50 healthy control subjects from Xuanwu Hospital of Capital Medical University between April 2010 and April 2012 were included into this cross-sectional study.Neuropsychological tests was conducted after the disease had been stable for 8 weeks, including information processing speed, memory, executive functions, language and visual perception.Correlation between information processing speed and depression, fatigue, Expanded Disability Status Scale (EDSS) were studied. RESULTS: (1)MS patient groups demonstrated cognitive deficits compared to healthy controls.The Symbol Digit Modalities Test (SDMT) (control group 57±12; RRMS group 46±17; SPMS group 35±10, P<0.05) and Paced Auditory Serial Addition Task (PASAT) (control group 85±18; RRMS group 77±20; SPMS group 57±20, P<0.05) impaired most.SPMS patients were more affected compared to patients with RRMS subtypes, and these differences were attenuated after control for physical disability level as measured by the EDSS scores.MS patients, especially SPMS subtype, were more severely impaired than control group in the verbal learning test, verbal fluency, Stroop C test planning time, while visual-spatial function and visual memory were relatively reserved in MS patients.(2) According to the Pearson univariate correlation analysis, age, depression, EDSS scores and fatigue were related with PASAT and SDMT tests (r=-0.41--0.61, P<0.05). Depression significantly affected the speed of information processing (P<0.05). CONCLUSIONS: Impairment of information processing speed, verbal memory and executive functioning are seen in MS patients, especially in SPMS subtype, while visual-spatial function is relatively reserved.Age, white matter change scales, EDSS scores, depression are negatively associated with information processing speed.

Higher mortality in patients with right hemispheric intracerebral haemorrhage: INTERACT1 and 2.

BACKGROUND AND PURPOSE: Controversy exists over the prognostic significance of the affected hemisphere in stroke. We aimed to determine the relationship between laterality of acute intracerebral haemorrhage (ICH) and poor clinical outcomes. METHODS: A subsidiary analysis of the INTERACT Pilot and INTERACT2 studies--randomised controlled trials of patients with spontaneous acute ICH with elevated systolic blood pressure (BP), randomly assigned to intensive (target systolic BP <140 mm Hg) or guideline-based (<180 mm Hg) BP management. Outcomes were the combined and separate end points of death and major disability (modified Rankin scale (mRS) scores of 3-6, 6 and 3-5, respectively) at 90 days. RESULTS: A total of 2708 patients had supratentorial/hemispheric ICH and information on mRS at 90 days. Patients with right hemispheric ICH (1327, 49%) had a higher risk of death at 90 days compared to those with left hemispheric ICH after adjustment for potential confounding variables (OR, 1.77 (95% CI 1.33 to 2.37)). There were no differences between patients with right and left hemispheric ICH regarding the combined end point of death or major disability or major disability in the multivariable-adjusted models (1.07 (0.89 to 1.29) and 0.85 (0.72 to 1.01), respectively). CONCLUSIONS: Right hemispheric lesion was associated with increased risk of death in patients with acute ICH. The laterality of the ICH does not appear to affect the level of disability in survivors. TRIAL REGISTRATION NUMBER: URL: http://www.clinicaltrials.gov. Unique identifier: NCT00226096 and NCT00716079.

Role of Mael in early oogenesis and during germ-cell differentiation from embryonic stem cells in mice in vitro.

In a previous study, we have identified a set of conserved spermatogenic genes whose expression is restricted to testis and ovary and that are developmentally regulated. One of these genes, the transcription factor Mael, has been reported to play an essential role in mouse spermatogenesis. Nevertheless, the role of Mael in mouse oogenesis has not been defined. In order to analyse the role of Mael in mouse oogenesis, the expression of this gene was blocked during early oogenesis in mouse in vitro using RNAi technology. In addition, the role of Mael during differentiation of embryonic stem cells (ESC) into germ cells in vitro was analysed. Results show that downregulation of Mael by a specific short interfering RNA disrupted fetal oocyte growth and differentiation in fetal ovary explants in culture and the expression of several germ-cell markers in ESC during their differentiation. These results suggest that there is an important role for Mael in early oogenesis and during germ-cell differentiation from embryonic stem cells in mouse in vitro.

Griffipavixanthone from Garcinia oblongifolia champ induces cell apoptosis in human non-small-cell lung cancer H520 cells in vitro.

Griffipavixanthone (GPX) is a dimeric xanthone which was isolated in a systematic investigation of Garcinia oblongifolia Champ. In this study, we investigate the effect of GPX on cell proliferation and apoptosis on human Non-small-cell lung cancer (NSCLC) cells in vitro and determine the mechanisms of its action. GPX inhibited the growth of H520 cells in dose- and time-dependent manners, with IC50 values of 3.03 ± 0.21 µM at 48 h. The morphologic characteristics of apoptosis and apoptotic bodies were observed by fluorescence microscope and transmission electron microscope. In addition, Annexin V/PI double staining assay revealed that cells in early stage of apoptosis were significantly increased upon GPX treatment dose-dependently. Rh123 staining assay indicated that GPX reduced the mitochondrial membrane potential. DCFH-DA staining revealed that intracellular ROS increased with GPX treatment. Moreover, GPX cleaved and activated caspase-3. In summary, this study showed that GPX inhibited H520 cell proliferation in dose- and time-dependent manner. Further mechanistic study indicated that GPX induced cell apoptosis through mitochondrial apoptotic pathway accompanying with ROS production. Our results demonstrate the potential application of GPX as an anti-non-small cell lung cancer agent.