Objective:To investigate the effect of β-elemene on mitochondrial structure and function of the A549 cells of non-small cell lung cancer(NSCLC),and to elucidate the mechanism of β-elemene in the treatment of NSCLC.Methods:The A549 cells at logarithmic growth stage were divided into blank control group(0 mng·L-1 β-elemene),low,medium and high doses of β-elemene groups(10,25 and 50 mg·L-1),and solvent control group(0.5％ethanol in equal volume).After treatment for 24 h,the cell activities in various groups were detected by MTT assay;the morphology changes of mitochondria in the cells in various groups was observed by transmission electron microscope;the levels of adenosine 5′-triphosphate(ATP)in the cells in various groups were detected by colorimetry;the mitochondrial membrane potential of the A549 cells in various groups were detected by JC-1 flow cytometry;mitochondrial membrane permeability transfer hole assay was used to detect the mitochondrial membrane permeabilities of the cells in various groups.Results:The MTT results showed that compared with blank control group,the cell activities in low,medium and high doses of β-elemene groups were decreased gradually(P＜0.05),while the cell activity in solvent control group had no significant change,and the difference was not significant(P＞0.05).The transmission electron microscope results showed that compared with blank control group,the mitochondria of A549 cells in low,medium and high doses ofβ-elemene groups showed swelling,vacuolation,disordered arrangement and dissolution,while the mitochondrial morphology of the A549 cells in solvent control group had no significant changes.The colorimetric method results showed that compared with blank control group,the ATP levels in the A549 cells in low,medium and high dose β-elemene groups were gradually decreased(P＜0.05),while the ATP level in the A549 cells in solvent control group had no significant change,and the difference was not significant(P＞0.05).The JC-1 flow cytometry method results showed that compared with blank control group,the mitochondrial membrane potential of the A549 cells in low,medium and high doses ofβ-elemene groups were decreased,and the percentages of the cells in Q2-4 region were increased(P＜0.05);the percentage of the A549 cells in the Q2-4 region in solvent control group had no significant change.The results of mitochondrial membrane permeability transfer hole experiment showed that compared with blank control group,the mitochondrial membrane permeabilities of the A549 cells in low,medium and high doses of β-elemene groups were increased,and the percentages of the cells in M4 region were increased(P＜0.05);the mitochondrial membrane permeability of the A549 cells and the percentage of the M4 cells in solvent control group had no significant changes,and the difference was not significant(P＞0.05).Conclusion:β-elemene can inhibit the proliferation of the A549 cells,and the mechanism may be that the mitochondrial structure of A549 cells is damaged by reducing the level of ATP and mitochondrial membrane potential,changing the mitochondrial morphology and increasing the mitochondrial membrane permeability.

Lysosomes represent a promising target for cancer therapy and reducing drug resistance. However, the short treatment time and low efficiency of lysosomal targeting have limited the application in lysosome-targeting anticancer drugs. In this study, we proposed an adhesive-bandage approach and synthesized a new lysosomal targeting drug, namely long-term lysosome-targeting anticancer drug (LLAD). It contains a SLC38A9-targeting covalently bound moiety and an alkaline component both to prolong the inhibition of SLC38A9 in lysosomes and alkalinize lysosomes. Upon short term and low-dose treatment of HeLa cells, at passage 0, with LLAD, it rapidly alkalinized lysosomes and also can be detected in lysosomes even at passage 15. LLAD induced apoptosis in HeLa cells through long-term lysosomal damage, and showed better long-term anticancer effect than cisplatin in vivo. Overall, our study paves the way for developing long-term lysosomal targeting drugs to treat cancer and overcome the drug resistance of cancer cells, and also provides a candidate drug, LLAD, for treating cancer.

Objective To explore the feasibility of long non-coding RNAs (lncRNAs) as biomarkers for radiation-induced intestinal injury. Methods Mice were exposed to 15 Gy of ⁶⁰Co γ-rays to the abdominal area. The pathological changes in intestinal tissues were analyzed at 72 h post-irradiation to confirm the successful establishment of the radiation-induced intestinal injury model. Real-time quantitative PCR was conducted to detect the expression of candidate radiation-responsive lncRNAs in the jejunum, jejunal crypts, colon tissues, and plasma of irradiated mice. Human intestinal epithelial cell line HIEC-6 and human colon epithelial cell line NCM460 were exposed to 0, 5, 10, and 15 Gy of ⁶⁰Co γ-rays. The expression levels of candidate lncRNAs were measured at 4, 24, 48, and 72 h post-irradiation to observe their changes with the irradiation dose. Results Pathological analysis showed that abdominal irradiation with 15 Gy successfully established an acute radiation-induced intestinal injury mouse model. Real-time quantitative PCR showed that Dino, Lncpint, Meg3, Dnm3os, Trp53cor1, Pvt1, and Neat1 were significantly upregulated following the occurrence of radiation-induced intestinal injury (P < 0.05). Among them, Meg3 and Dnm3os in mouse plasma were significantly upregulated (P < 0.05), while Gas5 was significantly downregulated (P < 0.05). In HIEC-6 and NCM460 cells, the expression levels of DINO, MEG3, DNM3OS, and GAS5 showed dose-dependent patterns at certain time points (P < 0.05). Conclusion The lncRNAs encoded by MEG3, DNM3OS, and GAS5 in intestinal epithelial cells are responsive to ionizing radiation. Consistent differential expression changes were detected in mouse plasma and intestinal tissues, indicating their potential as biomarkers for radiation-induced intestinal injury.

With the widespread application of ionizing radiation in many industries and the construction of nuclear power plants, the potentials for nuclear accidents is also increasing. In the event of a nuclear accident, rapid classification of a large population is generally involved, so accurate estimation of the radiation dose to the exposed population is the primary task of nuclear emergency response. Based on this need, World Health Organization and International Atomic Energy Agency have each established a worldwide network of biological dosimetry laboratories. In addition, regional networks of biological dosimetry laboratories have been established in the European Union, North America, Latin America and Asia. Based on the long-term organization of national training and assessment of biological dose estimation technology, China will also establish its own network of biological dosimetry laboratories in the future to cope with the emergency disposal needs of potential nuclear accidents. In this paper, the international biodosimetry network and related work will be reviewed, and the idea of establishing biodosimetry laboratory network in China will be elaborated.

With the widespread application of ionizing radiation in many industries and the construction of nuclear power plants, the potentials for nuclear accidents is also increasing. In the event of a nuclear accident, rapid classification of a large population is generally involved, so accurate estimation of the radiation dose to the exposed population is the primary task of nuclear emergency response. Based on this need, World Health Organization and International Atomic Energy Agency have each established a worldwide network of biological dosimetry laboratories. In addition, regional networks of biological dosimetry laboratories have been established in the European Union, North America, Latin America and Asia. Based on the long-term organization of national training and assessment of biological dose estimation technology, China will also establish its own network of biological dosimetry laboratories in the future to cope with the emergency disposal needs of potential nuclear accidents. In this paper, the international biodosimetry network and related work will be reviewed, and the idea of establishing biodosimetry laboratory network in China will be elaborated.

OBJECTIVE To investigate the serum toll-like receptor 4(TLR4),matrix metalloproteinase-3(MMP-3),in-terleukin-1β(IL-1β)and peripheral blood helper T-cell 17/regulatory T-cell(Th17/Treg)levels in patients with chronic periodontitis infected with porphyromonas gingivalis(Pg),as well as their diagnostic value of Pg infection in patients with chronic periodontitis.METHODS Totally 390 cases of patients with chronic periodontitis who at-tended Tianjin Stomatological Hospital from Jan.2022 to Jan.2024 were selected as the study subjects,and were divided into the infected group(113 cases)and the uninfected group(277 cases)according to Pg infection,and those patients in the infected group were further classified into the mild group(31 cases),the moderate group(45 cases),and the severe group(37 cases)according to the condition of the disease.Serum TLR4,MMP-3,IL-1β,and peripheral blood Th17/Treg levels were compared between the infected and uninfected groups and patients with different conditions of chronic periodontitis Pg infection,and the infected group was included in the positive and the uninfected group was included in the negative,and the receiver's operating characteristics(ROC)curves were plotted to obtain the area under the curve(AUC)to analyze the diagnostic value of single and combined de-tection of serum TLR4,MMP-3,and IL-1β,Peripheral blood Th17/Treg for Pg infection in patients with chronic periodontitis.RESULTS Serum TLR4,MMP-3,IL-1β and peripheral blood Th17/Treg levels in the infected group were(20.41±5.12)ng/ml,(6.12±2.03)μg/L,(10.24±2.19)μg/L and(0.32±0.10),higher than those in the uninfected group(P＜0.05).Serum TLR4,MMP-3,IL-1β and peripheral blood Th17/Treg levels in the se-vere group were(22.58±6.10)ng/ml,(8.12±2.67)μg/L,(12.49±3.10)μg/L and(0.52±0.15),higher than those in the moderate group[(14.78±3.44)ng/ml,(6.15±2.07)μg/L,(7.23±1.99)μg/L and(0.22±0.07)]and mild group(P＜0.05),which in the moderate group was higher than those in the mild group(P＜0.05).The results of ROC analysis showed that the AUC of serum TLR4,MMP-3,IL-1β,and peripheral blood Th17/Treg combined test for diagnosing Pg infection in patients with chronic periodontitis was 0.937,which was higher than that of the four single tests(P＜0.05).CONCLUSION Serum TLR4,MMP-3,IL-1β and peripheral blood Th17/Treg were highly expressed in chronic periodontitis patients infected with Pg,and all four indicators can participate in the progression of the disease,and the combined detection of these four indicators has a higher diagnostic value for Pg infection in patients with chronic periodontitis.

OBJECTIVE To investigate the serum toll-like receptor 4(TLR4),matrix metalloproteinase-3(MMP-3),in-terleukin-1β(IL-1β)and peripheral blood helper T-cell 17/regulatory T-cell(Th17/Treg)levels in patients with chronic periodontitis infected with porphyromonas gingivalis(Pg),as well as their diagnostic value of Pg infection in patients with chronic periodontitis.METHODS Totally 390 cases of patients with chronic periodontitis who at-tended Tianjin Stomatological Hospital from Jan.2022 to Jan.2024 were selected as the study subjects,and were divided into the infected group(113 cases)and the uninfected group(277 cases)according to Pg infection,and those patients in the infected group were further classified into the mild group(31 cases),the moderate group(45 cases),and the severe group(37 cases)according to the condition of the disease.Serum TLR4,MMP-3,IL-1β,and peripheral blood Th17/Treg levels were compared between the infected and uninfected groups and patients with different conditions of chronic periodontitis Pg infection,and the infected group was included in the positive and the uninfected group was included in the negative,and the receiver's operating characteristics(ROC)curves were plotted to obtain the area under the curve(AUC)to analyze the diagnostic value of single and combined de-tection of serum TLR4,MMP-3,and IL-1β,Peripheral blood Th17/Treg for Pg infection in patients with chronic periodontitis.RESULTS Serum TLR4,MMP-3,IL-1β and peripheral blood Th17/Treg levels in the infected group were(20.41±5.12)ng/ml,(6.12±2.03)μg/L,(10.24±2.19)μg/L and(0.32±0.10),higher than those in the uninfected group(P＜0.05).Serum TLR4,MMP-3,IL-1β and peripheral blood Th17/Treg levels in the se-vere group were(22.58±6.10)ng/ml,(8.12±2.67)μg/L,(12.49±3.10)μg/L and(0.52±0.15),higher than those in the moderate group[(14.78±3.44)ng/ml,(6.15±2.07)μg/L,(7.23±1.99)μg/L and(0.22±0.07)]and mild group(P＜0.05),which in the moderate group was higher than those in the mild group(P＜0.05).The results of ROC analysis showed that the AUC of serum TLR4,MMP-3,IL-1β,and peripheral blood Th17/Treg combined test for diagnosing Pg infection in patients with chronic periodontitis was 0.937,which was higher than that of the four single tests(P＜0.05).CONCLUSION Serum TLR4,MMP-3,IL-1β and peripheral blood Th17/Treg were highly expressed in chronic periodontitis patients infected with Pg,and all four indicators can participate in the progression of the disease,and the combined detection of these four indicators has a higher diagnostic value for Pg infection in patients with chronic periodontitis.

Objective:To explore classification models for radiosensitive lipid metabolites in human peripheral blood by combining lipidomics with multiple machine learning (ML) algorithms.Methods:Totally 97 peripheral blood samples were collected from 25 leukemia cases admitted to a general hospital in Beijing from March to September 2023 who were ready to undergo bone marrow transplantation, including 0 Gy blood samples before irradiation in the control group ( n=24), and 73 blood samples after irradiation at doses of 4, 8 and 12 Gy in the radiation group ( n=73), and the targeted lipidomic based on the ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) platform method to analyze the differences of different lipids between control and radiation groups. Then, lipids responsive to radiation doses of 0-12 Gy were identified using linear regression. Finally, classification models were constructed using five ML algorithms based on the training set, followed by the validation and evaluation of these models using the validation set. Results:Compared with the control group, the differences in the concentration changes of 62 lipids in 9 classes of lipid metabolites sensitive to radiation group were statistically significant ( t=-4.91 to 4.74, P<0.05), including sphingomyelins(SMs), cholesteryl esters(CEs), ceramides(Cers), phosphatidylinositols(PIs), hexosylceramides(HexCers), lysophosphatidylcholines (LysoPCs), phosphatidylcholines (PCOs), phosphatidylethanolamines (PEs), and lysophosphatidylethanolamines (LysoPEs). Twenty lipids responsive to radiation doses of 0-12 Gy were identified, namely 11 SMs, 7 CEs, 1 Cer, and 1 PI. The five models based on ML algorithms of decision tree (DT), support vector machine (SVM), light gradient boosting machine (Light GBM), random forest (RF), and K-nearest neighbors (KNN) all exhibited high goodness of fit (F1=0.69-1.00) and high sensitivity. The evaluation and validation metrics revealed that the RF-based model yielded the optimal radiation classification discrimination (sensitivity: 1.00; accuracy: 0.72; F1 score: 0.80). Conclusions:Lipid metabolites responsive to radiation and lipids responsive to radiation dose in human samples were identified using targeted lipidomics. The RF-based model can provide new ideas for exploring models of human radiosensitive lipid metabolites.

Objective To explore the effect of hyperlipidemia and lipid-lowering therapy on the prognosis of postoperative patients with hepatitis B related hepatocellular carcinoma.Methods The clinical data of the patients with hepatitis B related hepatocellular carcinoma who were operated in our hospital from January 2012 to January 2021 were retrospectively collected.The effect of blood lipid level and related lipid-lowering therapy on the prognosis of postoperative patients with hepatitis B related hepatocellular carcinoma was analyzed.Results Among 166 patients with hepatitis B related hepatocellular carcinoma,there were 63 cases had hyperlipidemia,of which 33 cases were treated by statins.The median postoperative disease free survival time in the hyperlipidemia group was significantly lower than that in the normal blood lipid group(24.8 months vs.38.5 months,P＜0.05),and the median overall survival time in the hyperlipidemia group was also significantly lower than that in the normal blood lipid group(30.1 months vs.44.5 months,P＜0.05).There was no statistically significant difference in prognosis between the patients with hyperlipidemia who used statins or not.The median disease free survival time was 23.4 months vs.26.3 months,and the median overall survival time was 29.7 months vs.30.3 months.Conclusions Hyperlipidemia is a risk factor for disease free survival and overall survival after surgery in the patients with hepatitis B related hepatocellular carcinoma.The use of statins alone in hyperlipidemia patients cannot reduce the risk of recurrence and prolong survival time.

Objective:To explore the changes of oxidative stress(OS),DNA damage and the occurrence of cellular premature aging of human immortalized keratinocytes(HaCaT)after that was radiated by X-ray with different doses.Methods:HaCaT cells were radiated by X-ray,and they were divided into 0 Gy group,5 Gy group and 10 Gy group according to the irradiation dose.The levels of intracellular reactive oxygen species(ROS)were detected by 2,7-Dichlorofluorescein diacetate(DCFH-DA)fluorescent probe,and the intracellular content of malondialdehyde(MDA)of lipid peroxidation products and the activity of superoxide dismutase(SOD)were measured by colorimetry.Immunofluorescence staining was used to detect the phosphorylated histone 2A variant(γ-H2AX)in HaCaT cells that were radiated by X-ray with different doses.Cell count kit-8(CCK-8)was used to detect the effect of X-ray with different doses on the proliferation of HaCaT cells after X-ray with different doses radiated them.β-Galactosidase staining was used to detect the proportion of premature aging cells.The changes of p21 and p53 protein expressions after X-ray irradiation were detected by Western blot.Results:After HaCaT cells were radiated by X-ray for 24h,the fluorescence intensity of 2',7'-Dichlorofluorescein(DCF)in 5 Gy and 10 Gy groups were significantly higher than that in the 0 Gy group,and the MDA contents of them were significantly higher than that in the control group,and the SOD activities of them were significantly lower than that in the control group(F=38.35,92.22,5.22,P<0.05),respectively.The change of γ-H2AX focus showed a dose-dependent significant increase at 1 h after irradiation,and the difference between them and control group was statistically significant(F=129.3,P<0.05).At 6h,24h and 48h after X-ray radiated HaCaT cells,the cell proliferation abilities of 5 Gy group and 10 Gy group were significantly decreased than that of 0 Gy group(F=116.41,62.20,34.29,P<0.01),and the β-Galactosidase activity of the two groups were significantly increased than that of 0 Gy group,and the difference was significant(F=1629.22,P<0.01).At 72h after X-ray with different doses radiated HaCaT cells,the expression levels of p21 and p53 proteins of 5 Gy group and 10 Gy group increased,and the differences of them among three groups were significant(F=104.4,66.69,P<0.01),respectively.Conclusion:Ionizing radiation can induce the occurrences of oxidative stress and DNA damage in HaCaT cells,and cause the occurrence of cellular premature aging.