OBJECTIVE: To investigate mechanisms of antioxidant and anti-inflammatory effects mediated by Irisin in mouse macrophages RAW264.7. METHODS: RAW264.7 cells were treated with 200 ng/mL Irisin for 0, 24 and 48 h. Real-time PCR was used to detect mRNA expressions of anti-inflammatory factors (IL-10,Arg-1,and CD206),antioxidant genes (HO-1 and SOD2) and PPAR-γ. ELISA assay was used to detect IL-10, Arg-1 in cell supernatants. Commercial kits were used to determine antioxidant enzyme activities,and Western-blot and immunofluorescence to detect Nrf2 and PPAR-γ expression and localization. Furthermore, after 24 h with 100 ng/mL LPS stimulation of RAW264.7 and 4 h 200 ng/mL Irisin pretreatment, real-time PCR was used to detect pro-inflammatory cytokines. DCFH-DA and Mito-LX were used to detect ROS levels. PPAR-γ inhibitor T0070907 (30 μmol/mL) was pretreated for 4 h and 200 ng/mL Irisin for 24 h, real-time PCR was used to detect expressions of anti-inflammatory cytokines. RESULTS: Compared with the untreated cells,Irisin promoted the expression and release of anti-inflammatory factors such as IL-10, Arg-1 and CD206 (P<0.05). However, Irisin inhibited LPS-induced TNF-α and IL-6 mRNA expression (P<0.05), which indicated that Irisin has anti-inflammatory effects. In addition, Nrf2 nuclear translocation occurred after irisin treatment in RAW264.7 cells. Expressions and enzyme activities of HO-1 and SOD2 were significantly enhanced, and the contents of GSH increased (P<0.05). In response to Irisin stimulation, PPAR-γ expression was elevated and underwent nuclear translocation, while PPAR-γ inhibitor T0070907 blocked the Irisin-induced anti-inflammatory response (P<0.05). CONCLUSION: Irisin initiated macrophage M2-type polarization and anti-inflammatory responses through enhancement of PPAR-γ-related anti-inflammatory genes and Nrf2-related antioxidant enzymes. Therefore,Irisin may be a candidate drug for inflammation-related diseases.

OBJECTIVE: To investigate the role and mechanism of IRAK4 in acute liver cell injury induced by acetaminophen (APAP). METHODS: Human liver cell line L02 cells were divided into control and treatment groups. The latter were treated with 20 mmol/L APAP for 6,12 and 24 hours. Cell viability was detected by CCK-8 method,apoptosis rates were detected by flow cytometry combined with AnnexinV/PI double staining, mitochondrial changes were detected by MitoSOX and JC-1 fluorescence dye, relative expression levels of IRAK4 were detected by fluorescence quantitative PCR (qPCR) and Western blot,L02 cell viability, and apoptosis and mitochondrial reactive oxygen species (mtROS) were detected after silencing the IRAK4 gene. After treatment with 10 mmol/L N-acetylhemionine (NAC) for 24 hours,L02 cell viability was detected by CCK-8 method,and IRAK4 gene expression level was detected by qPCR. RESULTS: Compared with the control group,cell viabilities were reduced and apoptosis rates were significantly increased after the APAP treatment for 24 h (P<0.05). After 12 and 24 h treatments,the levels of mtROS increased significantly (P<0.05);mitochondrial membrane potential decreased significantly at different time points (P<0.05). qPCR and Western blot results showed that IRAK4 expression increased after treatment for 24 h (P<0.05). After silencing IRAK4 gene,compared with the control and APAP groups,cell viability increased,apoptosis decreased and mtROS decreased (P<0.05). Compared with APAP group,NAC treatment increased cell viability and decreased mRNA expression of IRAK4 gene (P<0.05). CONCLUSION: IRAK4 might be involved in APAP-induced acute liver cell injury and would be a therapeutic target of APAP liver cell injury.

OBJECTIVE: To identify differentially expressed microRNA(miRNA) and hub gene associated with diffuse large B-cell lymphoma (DLBCL) and to find novel targets for mechanistic studies of DLBCL. METHODS: Dataset GSE117063 was chosen from the Gene Expression Omnibus (GEO) database, which included plasma samples from 17 DLBCL patients and 14 healthy individuals,and DE-miRNAs were selected by GEO2R. The potential target genes were predicted by miRTarBase. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were implemented using Database for Annotation, Visualization and Integrated Discovery (DAVID). Additionally,hub genes were screened by the Protein-Protein Interaction (PPI) network and miRNA-gene regulatory network was constructed. Finally,differential expressions of hub genes were verified by Gene Expression Profiling Interactive Analysis (GEPIA). RESULTS: The up-regulation of hsa-miRNA-326 and down-regulation of hsa-miRNA-375 were significantly and differentially expressed in DLBCL and healthy tissues. The target genes mainly participated in positive regulation of cell proliferation and enriched in HTLV-I infection,PI3K-Akt signaling pathway,proteoglycans in cancer and so on. Seven regulated hub genes (AKT1,CCND1,ERBB2,TP53,MYC,CTNNB1 and HSP90AA1) were verified to be differentially expressed in DLBCL. CONCLUSION: The up-regulated miRNA-326 and down-regulaed miRNA-375 and target genes might play important roles in pathogenesis of DLBCL.

OBJECTIVE: To investigate incidence characteristics and trends of stomach cancer in the Nanshan District of Shenzhen city from 2011 to 2019, and to provide bases for prevention and control strategies of gastric cancer. METHODS: Based on the data of stomach cancer which were collected by the Shenzhen Chronic Disease Prevention and Management Office,the incidence and age-standardized rates (ASR) of stomach cancers were calculated. The annual percent change (APC) was used to evaluate the trend of stomach cancer in the District. RESULTS: From 2011 to 2019,there were 498 new cases of stomach cancer. The incidence rate was 7.21×10^-5,the ASR China was 1.399×10^-4,and the ASR world was 1.518×10^-4. The incidence rate of male was 8.35×10^-5,the age-standardized incidence rates for standard Chinese population and for world population were 1.809×10^-4 and 2.090×10^-4,respectively. The incidence rate of female was 5.99×10^-5, the age-standardized incidence rates for standard Chinese population and for world population were 11.71/10⁵ and 1.316×10^-4,respectively. The incidence for male was significantly higher. In terms of age distribution,the incidence began to rise rapidly after the age of 45 and reached a peak at the age group of 80-85 years old, when the incidence was 9.087×10^-4. Over time,the incidence of stomach cancer was stable. CONCLUSION: The incidence of stomach cancer in the Nanshan District of Shenzhen City was stable. The data indicate that strengthening prevention and control on stomach cancer in high-risk male groups over 45 years old is needed.

OBJECTIVE: To investigate relationships between expression of miR-101 gene cluster and risk of esophageal cancer. METHODS: Expression levels of the miR-101 gene cluster was analyzed using high-throughput detection miRNA gene chips and was verified using esophageal cancer RNA sequencing data in the cancer genome atlas (TCGA) database. In addition,total RNA was extracted from 33 patients' esophageal cancer tissues and adjacent tissues. These RNA samples were used to detect expression of the miR-101 gene cluster using real-time fluorescent quantitative PCR (qPCR). Relationships between abnormal expression of miRNA and risk of esophageal cancer were analyzed by logistic regression. The diagnostic efficacy of miR-101 gene cluster on esophageal cancer was evaluated using receiver operator characteristics curve (ROC). The Fisher test was used to analyze correlations between expression of miRNA in gene cluster and clinicopathological factors in the patients. RESULTS: High-throughput detection of miRNA chip and bioinformatics analyses of TCGA database showed that the expressions of mir-101-3p, mir-125a-5p and mir-145-5p were down regulated in esophageal carcinomas compared with normal esophageal tissues (P<0.05). qPCR results showed that expressions of the miR-101 gene cluster were low in esophageal cancers. Multivariate logistic regression analyses showed that expression levels of mir-101-3p, mir-127-5p and mir- 145-5p were negatively correlated with the risk of esophageal cancer[OR values were 0.717 (0.563,0.912),0.717 (0.534,0.962) and 0.597 (0.426,0.838),P< 0.05]. ROC analyses showed that AUCs of the miR-101 gene cluster was 0.753, 0.792, 0.763 and 0.800, respectively (P<0.05). In addition,expression levels of mir-101-3p and mir-145-5p were correlated with tumor size (P<0.05),and that of mir-125a-5p were correlated with lymph node metastasis (P<0.05). CONCLUSION: Expression of the miR-101 gene cluster was low in esophageal cancer tissues compared to adjacent normal tissues. The expressions can be used as dpotential biomarker for the diagnosis of esophageal cancer but their functions and regulatory mechanisms in the cancer process need to be further studied.

OBJECTIVE: To investigate infiltrations of different subtypes of tumor-associated macrophages in primary esophageal small cell carcinomas (PESC),and their relationships with clinico-pathological parameters and prognosis. METHODS: Clinico-pathological data of PESC from January 2000 to December 2019 in Cancer Hospital of the Shantou University Medical College were analyzed retrospectively. Infiltrations of the CD68 and CD163 macrophages were evaluated by immunohistochemistry. The Chi-square and Fisher's exact tests were used to evaluate their relationships with the clinico-pathological indices. Furthermore,Kaplan-meier and Multivariate Cox regression were used to analyze prognosis for PESC. RESULTS: CD68- and CD163-stained macrophages were located in cytoplasmic or membrane of tumor cells. CD68 macrophage infiltrations,specifically,were correlated with tumor T stages (χ²=6.336,r=-0.303,P=0.012). However,the infiltrations showed no association with the other clinico-pathological variables. Kaplan-meier analyses showed that the pTNM stage, number of surgical lymph node dissection, N stage, adjuvant chemotherapy and infiltration were associated with overall survival (P<0.05). Multivariate Cox regression analyses showed that pTNM stage, the number of lymph node dissection and the infiltration were independent prognostic factors. High infiltrations were significantly associated with overall survival compared with low infiltrations (Hazard ratio=0.340, 95% confidence interval 0.179-0.645; P=0.001). CONCLUSION: High infiltration of CD68 macrophages,but not CD163,was an independent prognostic factor for patients with PESC. Patients with high infiltrations had significantly favorable prognoses than those with low infiltration of CD68 macrophages.

OBJECTIVE: To study predictive values of preoperative laboratory indexes[derived neutrophil to lymphocyte ratio (dNLR),systemic immune inflammatory index (SII),and C-reactive protein (CRP)/albumin (ALB)] on recurrences and metastases in breast cancer patients. METHODS: The patients who received surgical treatment and diagnosed with breast cancer by postoperative pathology in Sichuan Neijiang Second People's Hospital from April 2016 to April 2019 were selected as the research objects. Their clinical and pathological data, followed-up recurrences and metastases and disease-free survival times was collected. The Kaplan Meier curve of disease-free survival was drawn. Relationships between different clinicopathological data and disease-free survival were analyzed by log-rank test. Influencing factors of disease-free survival were analyzed by COX regression model. Predictive values of different indices on disease-free survival were analyzed using ROC curve. RESULTS: Log rank test of Kaplan-Meier curve of disease-free survival showed that the cumulative disease-free survival rates of breast patients with different tumor stage, lymph node metastasis, neutrophils to lymphocyte ratio,platelet to lymphocyte ratio,lymphocyte to monocyte ratio,dNLR,SII,and CRP/ALB were statistically different (log-rank χ²=12.020,5.332,3.923,4.948,15.340,22.970,5.496,all P< 0.05). COX regression model analyses showed that AJCC stage,dNLR,SII,CRP/ALB were the influencing factors of disease-free survival of breast cancer patients (P<0.05). ROC curve analyses showed that dNLR,SII and CRP/ALB alone had predictive values for disease free survival. Regression equations of dNLR,SII and CRP/ALB were obtained in the logistic model, which could be used as a combined index to predict disease-free survival of breast cancer patients. CONCLUSION: Laboratory indices dNLR,SII and CRP/ALB were related to postoperative recurrences and metastases of breast cancers. The combination of the three indices can provide a basis for prognosis evaluations.

OBJECTIVE: To investigate significance of fibrinogen (FIB), fibrinogen to albumin ratio (FAR) and serum tumor markers in the diagnosis and clinical progression of esophageal squamous cell carcinoma (ESCC). METHODS: Sixty-six ESCC patients who were admitted to the Fourth Hospital of Hebei Medical University from April 2018 to October 2021 were selected as the ESCC group. Clinicopathological data and laboratory test indexes of these patients at initial diagnosis were collected. Clinicopathological data,blood routine, coagulation function, biochemistry, serum tumor markers[including Carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC-Ag)] and other laboratory test indicators were collected at the time of initial diagnosis of ESCC patients. Another 30 healthy patients without benign and malignant tumors and cardiovascular diseases were selected as the control group. The differences among indicators between the two groups were compared,and expression levels of different results in different TNM stages were analyzed, and ROC curves was made to evaluate the predictive value of FIB, FAR, CEA and SCC- Ag for clinical propression of EXCC. Correlations between these indicators and clinicopathological features were analyzed. RESULTS: FIB, FAR, ratios of fibrinogen to prealbumin (FPR), ratios of neutrophil to lymphocyte (NLR) NLR and ratios of platelets to lymphocytes (PLR) in the ESCC group were significantly higher than those in the control group (P<0.05). The levels of FIB, FAR, CEA and SCC-Ag in stage III-IV were significantly higher than those in stage I-II. Meanwhile, ROC curves showed that FIB, FAR, CEA and SCC-Ag had predictive values for the clinical progression of ESCC (P<0.05),and combined detection of the four had higher diagnostic value for the clinical progress of ESCC. N stage and TNM stage in high FIB,FAR and SCC-Ag groups (which is object to the cut off value of ROC curve) were significantly higher than those in the low FIB,FAR and SCC-Ag groups (P<0.05). The maximum tumor diameters and T stages of patients with high FAR values were also significantly higher than those of patients with low values (P<0.05). TNM stages of patients with high CEA values were significantly higher than that of patients with low values, and male patients were more than female patients (P<0.05). CONCLUSION: Levels of plasma FIB,FAR,and serum CEA,SCC-Ag were closely related to the diagnosis and clinical progress of ESCC. Compared with detection of tumor markers in serum alone,the combined detection of the four methods showed better clinical efficacy in differentiating the clinical progress of ESCC,therefore,they are worthy of clinical applications.

OBJECTIVE: To study effects of AZD1152 combined with ¹²⁵I particles on proliferations and apoptoses of triple negative breast cancer cells MDA-MB-231. METHODS: Cultured cells were divided into several groups; control, ¹²⁵I particle irradiation, 1 μmol/L AZD1152 treatment with 48 h, treated with ¹²⁵I particle irradiation and 1 μmol/L AZD1152 treated with 48 h. Cell proliferation inhibitory rates were detected using the MTT and cell viability using the CCK-8 assays. Cell ploidies,cell cycles and apoptoses were detected using flow cytometry with cells separately stained by propidium iodide (PI) and by Annexin V/PI double-staining. Expressions of apoptosis-related proteins (Bcl-XL, Bcl-2 and PARP), CyclinB1 and Phosphorylation levels of Histone H3 were analyzed using Western blotting. RESULTS: After treatments for 48 h, cell proliferation inhibitory rates were (0.61±0.32)%,(17.62±1.41)%,(29.67±0.41)%,(53.17±1.26)%,respectively. Cell viability rates were (94.88±0.22)%,(59.21±0.14)%,(42.05±0.17)% (32.12±0.36)%,respectively. The proportions of G2/M phase were (18.99±0.15)%,(38.05±0.23)%,(49.80±0.32)%,(75.52±0.45)%,respectively. The apoptosis rates of MDA-MB-231 cells were (17.48±0.24)%,(29.23±0.02)%,(63.11±0.27)%,with significantly differences in the different groups (P<0.05). Compared with (0.31 ±0.03)% in the control group, the observed increases were significant (P<0.05). Immunofluorescence and flow cytometry analyses showed that multinucleated and polyploid cells appeared in the 1μmol/L AZD1152 group in 48 hours,which were easy to form aneuploid cells. Compared with the irradiated,inhibitor and control groups,the combined irradiation with 1 μmol/L AZD1152 for 48 h effectively down-regulated the expressions of Bcl-2,p-Histone H3,CyclinB1 and promoted Bax and dissection of PARP (P<0.05). CONCLUSION: The combination of ¹²⁵I particles and AZD1152 treatments efficiently inhibited proliferation and promoted apoptosis in breast cancer cell line MDA-MB-231.

OBJECTIVE: To evaluate inhalation toxicity and safety of impurities CFC115 and HFC1243zf in the pharmaceutical HFA-134a as propellant in pharmaceutical metered-dose inhalers. METHODS: Cytotoxicity assay, Reverse mutation assay, acute inhalation toxicity, 7-day repeat dose inhalation irritation study,21-day repeat dose inhalation toxicity study and active systemic anaphylaxis of the mixture of CFC115 and HFC1243zf (7:3,V/V) were tested to provide a basis for HFA 134a to control the impurity limit of CFC115 and HFC1243zf. RESULTS: Result from acute inhalation studies demonstrated that the 4-hour LC₅₀ value was higher than 3 115 g/m³ for CFC115 and 832 g/m³ for HFC1243zf. Results from 7-day repeat dose inhalation irritation,active systemic anaphylaxis,Ames and cytotoxicity studies demonstrated that the mixture of CFC115 and HFC1243zf was not irritative, allergenic, mutagenic nor cytotoxic. 21-day repeat dose inhalation toxicity study were exposed, nose-only, to the mixture of CFC115 and HFC1243zf at levels of 3 205 g/m³ for CFC115 and 856 g/m³ for HFC1243zf 2 h/d,compared with the control group. Among these exposed rats,food intakes were decreased,body weight increased slowly. Values of Bas and Mon%,ALT, AST,ALP,T-BIL and UREA,and the urinary nitrite,ketone,PRO and LEU were mostly increased (P<0.05 or P<0.01). Brain coefficient values increased and ovarian wet weight values decreased. Histopathological changes of focal or multifocal endocardial necrosis and monocyte infiltration in 5 SD rats which had a treatment-related adverse effect. In addition,the lung wet weight and coefficient values of male rats decreased (P<0.01). CONCLUSION: In in vitro or short-term animal tests,the mixed gas of CFC115 and HFC1243zf showed no safety risk under high concentration of exposure or inhalation exposure. Long-term repeated high-concentration inhalation exposures showed a variety of toxic target organ effects in rats. However,these toxic concentrations were well above the clinical maximum exposure.

OBJECTIVE: To investigate sub-chronic toxicity of polyphenol and polysaccharide extracts from Areca catechu in Wistar rats. METHODS: A total of 100 Wistar rats were randomly divided into control group,and Areca polysaccharide polyphenols-treated groups:0.28,0.83 and 2.5 g/kg. From the 0.28 and 0.83 g/kg groups,20 in each group,were killed after 90 days of exposure. From the 2.50 g/kg and control groups (30 in each group),20 were killed after 90 days of exposure,and treatments were stopped for the remaining 10 which received normal feeding for the subsequent 28 days. Each treated rat received 10 mL/kg volume of continuous intragastric administration for 90 days,once a day,and the control rats received equal volume of distilled water. Changes of body weight, food availability, hematology, biochemical parameters, urinalysis, main organ coefficient and histopathological examination were detected. RESULTS: Total weight gain of the 2.50 g/kg group was lower than that of the control group during the treatment period. There were no significant differences in the food utilization rates between groups except that the male rats in the first week of 2.50 g/kg group was lower than that in the control group. Five male and five female rats in the 2.50 g/kg group showed symptoms of poor spirit,rough coat and sluggish reaction after 3 weeks of treatment. Two male and two female rats died during the treatment period. Flatulence and thinning of gastrointestinal wall were observed in naked rats,but no obvious abnormalities were found in the histopathological examination of major organs. There were no obvious abnormalities in other dose groups compared with the control group during the exposure and the recovery periods. Although there were statistical differences in hematology,blood biochemistry and individual urine indexes between the control and the other dose groups,they were all within the range of normal test values in our laboratory, and had no biological significance. The organ coefficient and histopathological examination of the main organs showed no obvious abnormalities. It was considered that the death of rats might have been caused by gastrointestinal flatulence due to intestinal fermentation of a large amount of polysaccharide. Under the conditions of this study,90-day oral treatment with 0.83 g/kg areca polysaccharide polyphenols was considered to be at the no observed adverse effect level (NOAEL). CONCLUSION: Oral administration of 2.50 g/kg areca polysaccharides polyphenols for 90 days caused lethargy and flatulence in Wistar rats. Although this symptom was reversible,excessive dosage caused mortality. Therefore,development and utilization of areca polysaccharides polyphenols should be controlled within the safe dose range.