OBJECTIVE: To investigate differential expression of long non-coding RNA (LncRNA) during glycidyl methacrylate (GMS)-induced malignant transformation of human bronchial epithelial 16HBE cells,and to explore the regulatory effect of LncRNA on cell function and related signaling pathways at different stages of malignant transformation. METHODS: 16HBE cells were divided into GMA treatment group and DMSO control group,and the cells of the two groups were collected respectively. LncRNA microarray was used to detect and analyze differences of LncRNA expression in the malignantly transformed 16HBE cells. The key differentially expressed LncRNAs were screened out using TargetScan and MiRanda databases. Cytoscape was used to construct a ceRNA regulatory network with differentially expressed LncRNAs as the core. After that,the target genes and pathways of differentially expressed LncRNA were analyzed and predicted,and the relative expression levels of the above LncRNA were detected and verified by real-time fluorescence quantitative PCR. RESULTS: Microarray screening results showed that,compared with DMSO group,16 LncRNAs in GMA group were up-regulated in the process of the malignant transformation. On this basis,a LncRNA-related ceRNA regulatory network consisting of 3 LncRNAs,32 mRNAs and 204 miRNAs was constructed. Three regulatory axes in the network were significantly correlated with GMA-induced malignant transformation of 16HBE cells (P<0.05),and this showed that LncRNA G013234-hsa-miR-378b-TMEM129,LncRNA CTA-384D8.35-hsa-miR-486-3p-LYPD,LncRNA G087116-miR-29b-3p-NAV1 were closely associated with the malignant transformation process ceRNA triples. Results of qPCR showed that relative expression levels of LncRNA G013234,CTA-384D8.35 and G087116 in different cell stages were consistent with results of LncRNA microarray. CONCLUSION: During GMA-induced malignant transformation of 16HBE cells,LncRNA G013234,CTA-384D8.35 and G087116 were up-regulated at different stages of transformation. Construction of the ceRNA regulatory network and key mRNAs related to cell malignant transformation formed the theoretical basis for the process of GMA-induced malignant transformation of 16HBE cells.

OBJECTIVE: To investigate main active ingredients and mechanisms of licorice and candied ginger decoction (LDGD) for breast cancers by network pharmacology and molecular docking. METHODS: Main effective components of licorice and dried ginger decoction were screened using TCMSP database. Genecards database was used to obtain disease targets for breast cancer. Protein interaction networks were constructed and analyzed using STRING database and Cytoscape3.6.1 software. GO enrichment analysis and KEGG pathway enrichment analysis were performed by DAVID database. Based on literature retrieval,the main active ingredients and key targets of LDGD were docked. RESULTS: The potential key targets of LDGD in the treatment of breast cancer included AKT1,MYC,TP53,EGF,etc. The biological processes of GO functional enrichment mainly included apoptosis signaling pathway,oxidative stress,etc. Cell components mainly included cyclin-dependent kinases,transcription factor complexes,serine/threonine protein kinase complexes,membrane regions,membrane microregions,etc. Molecular functions mainly included protein serine/threonine kinase activity,transcription factor activity,nuclear receptor activity,ubiquitin protein ligase binding,etc. KEGG pathway analyses showed that p53,TNF,IL-17,EGFR,apoptosis and other signaling pathways were mainly involved. KEGG pathway mainly involved p53,TNF,IL-17 signaling pathway,etc. AKT1 was the target and small molecule that bound to it were liquiritin,6-gingerol,6-shogaol,Liquiritigenin,Isoliquiritigenin and Isoliquiritin. The scoring values of liquiritin,6-gingerol,6-shogaol and liquiritigenin were 80% higher than that of the original ligand. CONCLUSION: Liquiritin,6-gingerol,6-shogaol and liquiritigenin might be the key anti-breast cancer components in LDGD,and the anti-cancer mechanisms might involve cell growth,apoptosis,oxidative stress,inflammation and other related signaling pathways.

OBJECTIVE: To compare serum immunoglobulin contents among different populations of residents in the downwind direction of a coking plant from 2015 to 2019 and to analyze changes in trends of abnormal rates. METHODS: In each summer from 2015 to 2019,residents from two villages in the downwind direction (1 to 2 km) of a coking plant in northern China were selected as the research group. They were recruited according to certain inclusion and exclusion criteria,and their blood and urine samples were collected to detect the contents of immunoglobulin (IgG,IgA and IgM) and urine creatinine (Cr) through an automatic biochemical analyzer. Abnormal rates of serum immunoglobulin contents were statistically analyzed. Contents of polycyclic aromatic hydrocarbon metabolite 1-hydroxypyrene in urine was detected by high performance liquid chromatography-fluorescence detection method. RESULTS: Among the research residents, the average serum IgG contents in women were higher than that of men for each year (P<0.01),and the average IgM contents were higher than that of men in 2015 and 2016 (P<0.05); The average serum IgG and IgA contents of adults were higher than those of minors in each year (P<0.01),while the average serum IgM content of adults was lower than that of minors in 2016 (P<0.01). In 2015-2019,the abnormal rates of serum immunoglobulin contents showed an overall upward trend and then a downward trend. There were significant differences in serum IgA and IgM between different years (P<0.01). And in 2016,the abnormal rates of serum IgG,IgA and IgM contents in adult were higher than those in minors (P<0.05). The abnormal rates of serum IgG contents in women were higher than that in men in 2015 and 2016 (P<0.05). In addition,from 2015 to 2019,the median detection levels of polycyclic aromatic hydrocarbon metabolite 1-hydroxypyrene in the urine of residents in this area were 0.468,0.797,0.678,0.169 and 0.021 μg/g,respectively. CONCLUSION: From 2015 to 2019,the serum immunoglobulin (IgG,IgA and IgM) levels of residents in the downwind direction of the coking plant showed certain gender and age differences,and their abnormal rates generally show an upward trend and then a downward trend.

OBJECTIVE: To investigate the injury effects of gatifloxacin in livers of mice and its possible mechanisms of action. METHODS: Thirty-two SPF male Kunming mice were selected as the research objects and randomly divided into 4 groups:normal (saline),low-dose (25 mg/kg),medium-dose (50 mg/kg) and high-dose (100 mg/kg). The administration volume was 10 mL/kg,and the intragastric administration was continued for 7 days. The normal group was given the corresponding volume of normal saline. Through the detection of alanine aminotransferase (ALT),aspartate aminotransferase (AST),alkaline phosphatase (AKP),creatinine (CRE) and triglyceride (TG) in the serum of each group of mice, the preliminary data indicated that gatifloxacin caused damage to liver tissues in mice. Therefore,the transcriptome sequencing technology was used to detect gene expression profiles in livers from each group,to screen differentially expressed genes,and to perform gene ontology (GO) functional classification of differential genes and Kyoto encyclopedia of genes and genomes (KEGG) signal pathway enrichment analysis. RESULTS: Compared with the normal group,the qualities of the livers from the high-dose group were significantly reduced (P<0.01),and the liver coefficients for the low- and medium-dose groups were significantly reduced (P<0.01). The serum ALT levels from the high-dose group were significantly reduced (P<0.01);the serum AST levels from the middle and high-dose groups were significantly reduced (P<0.01). Compared with the normal group,a total of 27 differentially expressed genes (including 20 up-regulated genes and 7 down-regulated genes) were screened out in the medium-dose group. GO functional classifications suggest that these genes were mainly enriched in 17 biological processes including the immune system,multicellular organisms,and multibiological processes. KEGG pathway analyses indicate that the differential genes were mainly enriched in 22 pathways including lipid metabolism,terpenoids and polyketide metabolism. CONCLUSION: Gatifloxacin caused alterations in liver functions of mice by affecting the balance of endocrine system such as bile acid and cholesterol and lipid metabolism in the liver of mice.

OBJECTIVE: To investigate protective effects of Sulfotanshinone Sodium on lung injury induced by environmental fine particulate matters(PM_2.5)in rats. METHODS: 24 clean male SD rats were randomly divided into 4 groups with 6 rats in each group:control group (normal saline),PM_2.5 group,tanshinone Ⅱ A sodium sulfonate group (STS group) and dexamethasone group (positive control). PM_2.5 suspension was administered by intratracheal instillation. Except the control group,the dose of the other three groups was 5.4mg/kg,once every three days,a total of 10 times. From the first day of exposure,the STS and dexamethasone groups were injected intraperitoneally at the concentration of 15 mg/kg and 0.5mg/kg respectively,once a day for 28 days. 24 hours after the last exposure,respiratory frequencies and enhanced pauses were measured by whole body plethysmography (WBP). Two days after the last exposure,2ml blood was collected through abdominal aorta,and the peripheral blood lymphocyte subsets were detected by flow cytometry. After the rats were killed,the left lung was lavaged,and the bronchoalveolar lavage fluid (BALF) was collected to determine the contents of interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the bronchoalveolar lavage fluid (BALF) by ELISA. Western blot was used to detect the expression of NF-κB protein in lung tissue. RESULTS: Compared with the control group,the respiratory f and Penh values of the model group were significantly increased (P<0.01);the CD4⁺ and CD4⁺/CD8⁺ values were decreased (P<0.05);the IL-1β,TNF-α and IL-6 values in BALF were increased (P<0.01),and the P65 contents in the lung cytoplasm were decreased while the p-P65 contents in the nucleus were increased (P<0.01). Compared with the model group,the f and Penh values in STS group decreased (P<0.05 or P<0.01);CD4⁺ and CD4⁺/CD8⁺ in peripheral blood increased (P<0.01 or P<0.05),and the contents of IL-1β,TNF-α,IL-6 in BALF were decreased (P<0.01 or P<0.05);P65 in cytoplasm increased and p-P65 content in nucleus decreased (P<0.01). CONCLUSION: STS had protective effects on rat lung inflammation induced by PM_2.5,via regulation of lung and immune functions.

OBJECTIVE: To investigate expression of Xklp2 target protein (TPX2) in non-muscle-invasive bladder cancer and its clinical significance. METHODS: Data from the GEPIA database were analyzed to confirm expression of TPX2 mRNA in bladder cancer and its relationship with survival of patients. In addition,immunohistochemical staining was also used to detect the expression of TPX2 protein in 60 non-muscle-invasive bladder cancer tissues. Their relationships with clinical pathological indicator of tumors and prognosis of patients were analyzed. RESULTS: By mining the GEPIA database,TPX2 mRNA expressions were up-regulated in bladder cancer,and patients with higher expressions had lower disease-free survival rate (P<0.05). Immunohistochemical staining results showed that positive expression rate of TPX2 protein was 75% (45/60),while no positive expression was found in the corresponding basal tissues. The difference between the two groups was statistically significant (P<0.01). Expression levels of TPX2 protein were negatively correlated with clinical pathological indicators of tumors such as tumor numbers,tumor diameters,pathological grades and postoperative recurrences (P<0.05). Survival analyses showed that bladder cancer patients with higher expression of TPX2 had shorter survival time (P<0.05). CONCLUSION: Expression of TPX2 was up-regulated in bladder cancer and its expression level was negatively correlated with prognosis of patients. Therefore,it has potential to become a tumor marker for predicting prognosis in bladder cancer patients.

OBJECTIVE: To investigate effects of metformin on proliferation and migration of human breast cancer cell MDA-MB-231 in vitro. METHODS: Cell cultures were divided into four treatment groups:RPMI 1640 medium group and metformin treatment groups (1,2,4 mmol/L). Cell viability was measured at 48 and 72 h after treatment. Morphological changes of cells were observed under inverted microscopes after cells were stained with Giemsa. Effects of metformin on cell migration were detected by wound healing and Transwell assays. RESULTS: The MTT assay showed that metformin at 4 mmol/L significantly inhibited cell viability l for 72 h,and the inhibition rate was (29.83±2.25%). Compared with the control group,the decrease of cell viability was statistically significant (P<0.05). In the 4 mmol/L treatment group,the number of round cells increased,the cell density decreased obviously,and the connection between cells decreased. After treatment with metformin at 4 mmol/L for 48 hours,the cells showed obvious nuclear fragmentation. Wound healing experiment showed that the cell confluence of 2,4 mmol/L treatment groups were obviously lower than that of the control group after 24 h. The wound healing rate of 4 mmol/L treatment group was (52.67±4.48%),which was significantly different from that of the control group (P<0.01). Transwell experiments showed that the number of perforating cells decreased after treated with metformin at the concentration of 2,4 mmol/L for 24 h. The numbers of perforating cells were (61.6±1.6) and (51.3±2.6),which was significantly different from that of the control group (99.3±18.9) (all P<0.05). CONCLUSION: Metformin inhibited proliferation and migration of human breast cancer cell in vitro.

OBJECTIVE: To investigate protective effects of a catalytic antioxidant AEOL-10150 against nitrogen mustard-induced liver injury and related mechanisms. METHODS: A single intraperitoneal injection of nitrogen mustard hydrochloride (2 mg/kg) was used to construct a nitrogen mustard-induced acute liver injury model in C57BL/6 mice. AEOL-10150 (5 mg/kg) was given 0.5 and 6 h for treatment intervention after nitrogen mustard exposure,and sacrificed after 3 days of exposure. For clarifying the AEOL-10150 has a protective effect on nitrogen mustard-induced liver injury in mice,the following indicators were evaluated:gross liver tissue changes and pathological changes by HE stain;serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities;liver tissue reactive oxygen species (ROS) level,malondialdehyde (MDA) content;reduced glutathione (GSH) content;liver tissue redox regulatory molecule Nrf-2 mRNA and protein expression level;liver tissue myeloperoxidase (MPO) activity,and serum inflammatory factors TNF-α,IL-1β content. RESULTS: A single intraperitoneal exposure of 2 mg/kg nitrogen mustard successfully induced oxidative damage and inflammatory response in livers of mice in the 3rd day. Compared with nitrogen mustard-exposed mice,the AEOL-10150 intervention reduced liver tissue pathological damage,decreased serum ALT and AST activities,and significantly reduced liver tissue ROS and MDA levels,accompanied by the increased GSH content and the decreased Nrf-2 level. In addition,the liver tissue MPO activities and serum TNF-α and IL-1β concentrations of mice in the AEOL-10150 intervention group were significantly lower than those in the nitrogen mustard-exposed group. CONCLUSION: AEOL-10150 effectively alleviated the acute liver injuries which were induced by nitrogen mustard in mice,and its mechanism might be related to direct antioxidant effect and anti-inflammatory effect.

OBJECTIVE: To explore the effects and mechanisms of action of a new polyoxometalate drug {BiW₈O₃₀} in liver cancer cells. METHODS: Human hepatocellular carcinoma cell line SK-HEP-1 was cultured and treated with 50,100,200 and 400 μmol/L {BiW₈O₃₀} solution,respectively. The control group was given normal culture medium. The corresponding indexes were detected after 24 and 48 h of culture. Cell proliferation assay was used to detect cell viability and half maximal inhibitory concentration (IC₅₀);cell colony formation experiment was used to detect number of colony formation;wound healing rate was detected by scratch test;Transwell assay was used to detect the number of invasive cells;the effect of drugs on apoptosis was analyzed by fluorescence staining. RESULTS: After treatment with 50,100,200,400 μmol/L {BiW₈O₃₀} for 24 and 48 h,the survival rate of SK-HEP-1 cells decreased by 29.98%,51.29%,65.81%,83.59% and 31.57%,66.23%,81.88%,83.97%,respectively (P<0.01),compared with the control group. In addition,IC₅₀ were (102.07±1.38) and (74.68±0.91) μmol/L,respectively. After 2 weeks of treatment with 50,100,and 200 μmol/L {BiW₈O₃₀},the number of colony formation of SK-HEP-1 cells decreased by 28.57%,63.27%,and 90.82%,respectively (P<0.01). After treatment with 50,100,200 μmol/L {BiW₈O₃₀} for 24 and 48 h,the scratch healing rate of SK-HEP-1 cells decreased by 8.47%,70.59%,80.83% and 19.56%,65.98%,75.49%,respectively (P<0.01),the number of invaded cells of SK-HEP-1 decreased by 27.74%,45.51%,68.77% and 47.03%,72.20%,84.07% (P<0.01). Observation after fluorescent staining revealed that the liver cancer cells treated with {BiW₈O₃₀} shrank,the nuclei shrank,fragmented,and apoptotic bodies appeared. CONCLUSION: The new polyoxometalate drug {BiW₈O₃₀} was toxic to SK-HEP-1 cancer cells in vitro by inhibiting the proliferation,colony formation,migration and invasion,and by inducing cell apoptosis.

OBJECTIVE: To construct and to evaluate activities of luciferase reporter plasmids for transcription factor STAT5A promoter. METHODS: The sequence for the promoter region in STAT5A gene was amplified by PCR using genomic DNA of the breast cancer cell line MCF7 as a template and cloned into luciferase reporter plasmid pGL3-Enhancer. The target plasmid was amplified by bacterial fluid,examined by enzyme digestion and direct sequencing. The function of target plasmid was further verified by dual luciferase reporter gene assay. RESULTS: It was found that the cloned sequence of STAT5A promoter region in target plasmid was correct,and the target plasmid had transcriptional activities. Dual-luciferase assays found that the relative activities of the recombinant vector pGL3-STAT5A-pro-luc-E luciferase were about 8 times that of the negative control pGL3-Enhancer (P<0.01),and the pGL3-STAT5A-pro-N-luc-E luciferase activities were about 6 times that of the negative control pGL3-Enhancer (P<0.01). CONCLUSION: A luciferase reporter plasmid for transcription factor STAT5A promoter was successfully constructed. The plasmids would provide an important research tool for further study of STAT signal transduction and the transcriptional activator protein signaling pathway.

OBJECTIVE: To investigate acute toxicity and genotoxicity of an innovative antitumor drug -serum thymus factor 9 peptide. METHODS: Mice,rats,beagles were given 70.0 mg/kg by one-time injection to test for acute toxicity. Ames test,CHL test and micronucleus test were used to investigate genotoxicity. RESULTS: After treatments,the animals were in good spirits and behaved normally. There were no signs of excitation or inhibition. No acute toxicity was found. Ames test showed no significant increase in the number of mutant colonies at 1-5 000 μg/dish,similar to the solvent control group. CHL test showed no significant increase in chromosome aberration rate at 1 400-5 600 μg/mL (less than 3%),there was no statistical significance compared with the solvent control group (P>0.05). The micronucleus cell rates in the 17.5-70.0 mg/kg group were all less than 2.0‰,and there was no statistical significance compared with the solvent control group (1.2‰) (P>0.05). CONCLUSION: The maximum tolerated dose of serum thymic factor 9 peptide was greater than 70.0 mg/kg,which was 930 times of the recommended dosage for clinical patients. Acute toxicity and genotoxicity were not observed in the dose ranges for this investigation.

OBJECTIVE: Studies on acute toxicity,repeated toxicity and teratogenicity of sodium sulfide after oral administration in rats. METHODS: SPF SD rats were selected,half male and half female. Horne's test was used to evaluate acute toxicity with dosing series of 215,464,1 000,2 150 mg/kg sodium sulfide. To study sub-chromic oral toxicity,rats were administered with the doses of 30,60 and 120 mg/kg. Teratogenicities were measured by conventional teratogenicity test with continuous administration for 10 days with 30,60 and 120 mg/kg in the second trimester. RESULTS: In the oral administration of sodium sulfide,LD₅₀ value and 95% confidence intervals were 794 (584-1 080) mg/kg for female and 926 (636-1 350) mg/kg for male. In the teratogenicity test, the rate of stillbirth (1.75%) and the rate of deformity (5.88%) were significantly increased in the high-dose group of 120 mg/kg (P<0.05),compared with the control group. CONCLUSION: The LD₅₀ of female and male SD rats were 794 mg/kg and 926 mg/kg respectively. In the repeated toxicity test,pathological changes in digestive tract existed in the high-dose group of 120 mg/kg. The no observed adverse effect level (NOAEL) in repeated administration toxicity tests was 60 mg/kg. Suspicious positive results occurred in the teratogenic tests.

OBJECTIVE: According to the "OECD Guidelines for the testing of chemicals 473",the in vitro mammalian chromosomal aberration test was used to determine whether anatase titanium dioxide nanoparticles could induce chromosome aberrations was genotoxic or not. METHODS: The final concentrations of the test substance were set to be 0.007 8,0.031 2,0.125,0.5,and 2 mg/mL. In addition,a solvent control group and a positive control group were set up at the same time. The treatment included short-term treatment group with metabolic activation (+S9,4 h),short-term treatment group without metabolic activation (-S9,4 h) and continuous treatment group without metabolic activation (-S9,24 h). The cultured Chinese hamster lung cells (CHL) were exposed to anatase nano-titanium dioxide suspension,and the cells were harvested after a corresponding period of time,treated with hypotonic solution and then fixed. The cells were spread onto slides and stained. For each dose group,300 well-dispersed metaphase cells were selected for chromosomal aberration analysis. RESULTS: At the dose level of 0.007 8,0.031 2,0.125,0.5 and 2 mg/mL without metabolic activation,the induced chromosome aberration rates (excluding the gap) were 1.67%,1.33%,2.00%,1.00% and 2.33%,respectively. With metabolic activation,the rates were 1.33%,1.33%,2.00%,1.67% and 2.00%,respectively. After 24 hours treatment without metabolic activation,the rates were 1.33%,1.67%,2.00%,1.67% and 2.33%,respectively. Compared with the solvent control group,there was no statistically significant difference observed (P>0.05). CONCLUSION: Under the current test conditions,anatase titanium dioxide nanoparticles showed no evidence of genotoxic potential in the CHL at the tested dose levels of 0.007 8-2 mg/mL.