Map of benzoapyrene metabolites-DNA adducts in human bronchial epithelial-like cells: Based on chromatin immunoprecipitation followed by sequencing technology / 环境与职业医学

Background The active metabolite of benzo[a]pyrene (BaP), 7,8-dihydroxy-9,10-epoxybenzo[a]pyrene (BPDE), can form adducts with DNA, but the spectrum of BPDE-DNA adducts is unclear. Objective To identify the distribution of BPDE adduct sites and associated genes at the whole-genome level by chromatin immunoprecipitation followed by sequencing (ChIP-Seq), and serve as a basis for further exploring the toxicological mechanisms of BaP. Methods Human bronchial epithelial-like cells (16HBE) were cultured to the fourth generation inthe logarithmic growth phase. Cells were harvested and added to chromatin immunoprecipitation lysis buffer. The lysate was divided into experimental and control groups. The experimental group received a final concentration of 20 μmol·L−1 BPDE solution, while the control group received an equivalent volume of dimethyl sulfoxide solution. The cells were then incubated at 37 °C for 24 h. Chromatin fragments of 100-500 bp were obtained through sonication. BPDE-specific antibody (anti-BPDE 8E11) was used to enrich DNA fragments with BPDE adducts. High-throughput sequencing was conducted to detect BPDE adduct sites. The top 1000 peak sequences were subjected to motif analysis using MEME and DREME software. BPDE adduct target genes at the whole-genome level were annotated, and Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of BPDE adduct target genes were conducted using bioinformatics techniques. Results The high-throughput sequencing detected a total of 842 BPDE binding sites, distributed across various chromosomes. BPDE covalently bound to both coding and non-coding regions of genes, with 73.9% binding sites located in intergenic regions, 19.6% in intronic regions, and smaller proportions in upstream 2 kilobase, exonic, downstream 2 kilobase, and 5' untranslated regions. Regarding the top 1000 peak sequences, four reliable motifs were identified, revealing that sites rich in adenine (A) and guanine (G) were prone to binding. Through the enrichment analysis of binding sites, a total of 199 BPDE-adduct target genes were identified, with the majority located on chromosomes 1, 5, 7, 12, 17, and X. The GO analysis indicated that these target genes were mainly enriched in nucleic acid and protein binding, participating in the regulation of catalytic activity, transport activity, translation elongation factor activity, and playing important roles in cell division, differentiation, motility, substance transport, and information transfer. The KEGG analysis revealed that these target genes were primarily enriched in pathways related to cardiovascular diseases, cancer, and immune-inflammatory responses. Conclusion Using ChIP-Seq, 199 BPDE adduct target genes at genome-wide level are identified, impacting biological functions such as cell division, differentiation, motility, substance transport, and information transfer. These genes are closely associated with cardiovascular diseases, tumors, and immune-inflammatory responses.

7,8-dihydroxy-9,10-epoxybenzoapyrene induced ferroptosis in mouse hippocampal neuron HT22 cells and its mechanism / 环境与职业医学

Background Benzo[a]pyrene (BaP) has neurotoxicity, which can induce the loss of hippocampal neurons in humans and animals and lead to spatial learning and memory dysfunction, but its mechanism is still unclear. Objective To observe the ferroptosis of mouse hippocampal neuron HT22 cells induced by 7,8-dihydroxy-9,10-epoxybenzo[a]pyrene (BPDE), an active metabolite of BaP, and to explore its potential mechanism, so as to provide a basis for the study of BaP neurotoxicity mechanism. Method Mouse hippocampal neuron HT22 cells were selected and divided into four groups: solvent control group and low, medium, and high concentration BPDE exposure groups (0.25, 0.50, and 0.75 μmol·L−1). Cell survival was detected by CCK8 method. Cell morphology and ultrastructure were observed under light and electron microscopes. The levels of reactive oxygen species (ROS) and Fe2+ were detected by fluorescence probe method. Iron, 4-hydroxynonenoic acid (4-HNE), malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GSH-PX) levels were detected with commercial kits. The expression levels of acyl-CoA synthase long chain family member 4 (ACSL4), cyclooxygenase 2 (COX2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) were detected by Western blotting. After interventions with ferroptosis inhibitors 20 μmol·L−1 deferoxamine (DFO) and 10 μmol·L−1 ethyl 3-amino-4-cyclohexylaminobenzoate (Fer-1), the cell survival rate of each BPDE exposure group and the changes of the ferroptosis characteristic indicators and protein expression levels were observed. Results With the increase of BPDE concentration, the survival rate of HT22 cells decreased gradually, and the survival rate of each BPDE group was significantly lower than that of the solvent control group (P<0.01). Under light microscope, the number of cells in the high concentration BPDE group was significantly reduced, and atrophic cells and reduced synapses were recorded. Under electron microscope, the HT22 cells in the high concentration BPDE group showed mitochondrial shrinkage, decreased crista, and increased mitochondrial membrane density. Compared with the solvent control group, the levels of intracellular lipid ROS, Fe2+, 4-HNE, and MDA significantly increased in the high concentration group (P<0.01), the GSH and GSH-PX levels were significantly decreased (P<0.01), the protein expression levels of ASCL4 and COX2 were significantly increased (P<0.01), and the protein expression levels of SCL7A11 and GPX4 were significantly decreased (P<0.01). The ferroptosis inhibitors DFO and Fer-1 significantly reversed the cell survival rate (P<0.01), the ferroptosis characteristic indicators (ROS, Fe2+, 4-HNE, MDA, GSH, and GSH-PX levels) (P<0.01), and the expression levels of ferroptosis-related proteins (ACSL4, COX2, SLC7A11, and GPX4) (P<0.01) in the high concentration BPDE group. Conclusion BPDE can induce ferroptosis in mouse hippocampal neuron HT22 cells, which may be related to the inhibition of SLC7A11/GSH/GPX4 axis and the induction of iron metabolism disorder.

Effects of subchronic exposure to benzoɑpyrene on memory function and mRNA expressions of NMDA receptors in different brain regions in rats / 环境与职业医学

Background The altered expressions of hippocampal N-methyl-D-aspartate (NMDA) receptors induced by benzo[ɑ]pyrene (BaP) causes short-term spatial learning and memory impairment in humans and animals, but whether BaP causes alterations of NMDA receptor subunits in other brain regions and the associated neurotoxic mechanism is still essentially unknown. Objective To observe the mRNA expressions of NR1, NR2A, and NR2B of NMDA receptor subunits in different brain regions in SD rat model with subchronic exposure to BaP, and to provide a basis for in-depth study of the mechanism of BaP-induced neurotoxicity. Methods Forty male SD rats were selected and randomly divided into a control group and 1.00, 2.50, and 6.25 mg·kg−1 BaP exposure groups with 10 rats in each group. The exposure rats received intraperitoneal injection of BaP every other day for 90 d.The average latency to platform, the average total distance, and the duration spent in previous quadrant were measured by the Morris Water Maze. Real-time fluorescence quantitative PCR was used to detect the mRNA expressions of NR1, NR2A, and NR2B in hippocampus, cortex, cerebellum, and striatum of rats. Results The average latency to platform and the average total distance in the 2.50 and 6.25 mg·kg−1 BaP groups were significantly prolonged compared with the control group (P<0.05), and the duration that rats spent in previous quadrant in the 6.25 mg·kg−1 BaP group was significantly shortened (P<0.05). Compared with the control group, the mRNA expressions of NR1 and NR2B in the hippocampus in the 2.50 and 6.25 mg·kg−1 BaP groups were significantly reduced (P<0.05), and the NR2A mRNA expression in the hippocampus in the 6.25 mg·kg−1 BaP group was significantly reduced (P<0.05); the mRNA expressions of NR1 and NR2B in the cortical tissue in the 6.25 mg·kg−1 BaP group were significantly reduced (P<0.05), and the mRNA expression of NR2A in the cortical tissue in the 1.00 mg·kg−1 BaP group was reduced; the mRNA expression of NR2B in the cerebellar tissue in the 6.25 mg·kg−1 BaP group was significantly reduced (P<0.05); there were no differences in the mRNA expressions of NMDA receptor subunits in the striatum tissue (P>0.05). Conclusion Subchronic BaP exposure can cause short-term spatial learning and memory impairment in rats, which may be related to the down-regulation of mRNA expressions of NR1, NR2A, and NR2B in hippocampus, changes of mRNA expressions of NR1, NR2A, and NR2B in cortical area, and the down-regulation of NR2B mRNA expression in cerebellum.

Discussion on the sharing mechanism of cerebration of constructing the sharing platform for respiratory disease biobank / 中华医学科研管理杂志

Objective:Standardized sample resources and high-quality clinical big data are important resources for medical research, only through resource sharing can maximize its utilization.Which can be utilized to the max only through resource sharing.Methods:This paper attempts to explore the sharing mechanism of the resource sharing platform and proposes some aspects such as the platform construction background, management regulations, legal ethical system, data sharing principles, benefit distribution, etc.This article attempts to explore the sharing mechanism based on the resource sharing platform of the respiratory disease biobank, proposes the contents that should be included in the sharing mode.Detailed information including the platform construction background, management procedures, legal and ethical system, data sharing principles and benefit distribution should take into consideration in the operating mechanism of the platform.Results:Establishing a resource sharing platform matches the development of clinical research in China.The tailored sharing model which is suitable for the field of respiratory diseases will also guide the rapid development of clinical research.Conclusions:The construction of a respiratory disease biobank sharing platform is conducive to promoting the opening and sharing of biological samples and information resources in the context of big data.

Lipid peroxidation and anti-lipid peroxidation and the skin pigment in skin of C57BL/6 mice exposed to arsenic / 中华地方病学杂志

Objective To study the effect of lipid peroxidation and anti-lipid peroxidation and the pigment in skin of C57BL/6 mice exposed to arsenic.Methods Forty male C57BL/6 mice were divided into four groups via the random number table method,ten mice in each group,and the mice were fed ad libitum drinking water containing arsenic at 0,1,5 and 25 mg/L concentrations for a period of 6 weeks,respectively.Twelve days before the end of the experiment,procedure of depilation was performed to induce anagen of hair cycle.On the 30th day of experiment,the activity of superoxide dismutase (SOD) was determined by nitrite method,the activity of glutathione peroxidase (GSH-Px) was determined by dithiobisobenzoic acid method,the content of malondialdehyde (MDA) in skin of C57BL/6 mice was determined by thiobarbituric acid method.Melanin was measured by NaOH dissolution method.Results No significant difference was found in body weight and water intaken between groups (F =0.119,0.363,P ＞ 0.05).The activity of SOD [(16.00 ± 5.05),(13.96 ± 2.02),(10.46 ± 3.14) U/mg prot] in 1,5 and 25 mg/L arsenic groups were all significantly lower than that in control group [(20.36 ± 4.82) U/mg prot,P ＜ 0.05].GSH-Px activity in 1,5 and 25 mg/L arsenic groups [(98.14 ± 23.92),(87.18 ± 10.87),(53.56 ± 19.97) U/mg prot] were all significantly lower than that in control group [(119.34 ± 33.14) U/mg prot,P ＜ 0.05].While MDA levels in 5 and 25 mg/L arsenic groups [(9.09 ± 2.04),(11.48 ± 2.21) nmol/mg prot] were significantly higher than that of control group [(6.19 ± 0.56) nmol/mg prot,P ＜ 0.05] and that of 1 mg/L arsenic group [(6.52 ± 1.67) nmol/mg prot,P ＜ 0.05).No significant difference was found in melanin levels (P ＞ 0.05).Conclusions Lipid peroxidation and the decreasing of antioxidation may be one of the mechanisms of arsenic-induced skin damage.Arsenic-induced skin melanin content change is not found.

Correlations of nasal responses to leukotriene D4 and histamine nasal provocation with quality of life in allergic rhinitis

BACKGROUND: The symptoms of allergic rhinitis (AR) greatly affect the quality of life (QoL) in the patients with AR. The correlations of nasal response to leukotriene D4 (LTD4) and histamine nasal provocation with health related QoL in AR are not clear. OBJECTIVE: To evaluate the correlations of nasal response to LTD4 and histamine nasal challenge with QoL in AR. METHODS: Patients randomly underwent LTD4 and histamine nasal challenge tests, completed the rhinoconjunctivitis quality of life questionnaire (RQoLQ), and rating the symptom severity score (total symptom score 4, TSS4) in the previous week. The correlations between nasal challenge tests induced nasal responses and QoL in RQoLQ were analyzed. RESULTS: A total of 25 eligible AR patients enrolled and finished both LTD4 and histamine nasal challenge and completed the questionnaire of RQoLQ. Histamine nasal challenge induced sneezing, increased nasal resistant were correlated with most of the dimensions (general, practical, nasal, eye problems, and quality of sleep, p < 0.05), while LTD4 nasal challenge induced sneeze, increased nasal resistant only correlated with nasal and ocular problems. On the contrary, the severity of the sneeze assessed by TSS4, was not correlated with QoL, while the severity of rhinorrhea, congestion, and nasal pruritus were correlated with nasal and practical problems, and nasal congestion was also correlated with ocular problems (r = 0.60, p = 0.01). CONCLUSION: LTD4 and histamine nasal challenge induced nasal responses were correlated with different clinical symptoms severity and QoL, which can be used as a good diagnosis and evaluation methods for the management of AR.

Some aspects need to focus on in humanistic quality education of undergraduate clinical medical students / 中华医学教育探索杂志

The humanistic quality education of undergraduate clinical medical students is a system of education throughout the whole process of medical education.Some aspects include new career planning education,medical humanities quality courses,social practice,professional courses,clinical practice and evaluation of humanistic quality education need to design and control,which are necessary to achieve a good education results.

Relationship between HSP70 gene polymorphisms and susceptibility to lung function injury of cock-oven workers / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>In this study, we investigate the relationship between HSP70 and lung function injury. To study on the feasibility of HSP70 genes polymorphisms as biological marker of the damage of pulmonary dysfunction susceptibility.</p><p><b>METHODS</b>183 cock-oven workers were selected as exposure groups and 143 workers unexposed workers were selected as control groups. We investigated their general information with uniform questionnaire. Pulmonary dysfunction indicators were determined using portable spirometer. HSP70-1 G190C, HSP70-2 A1267G, HSP70- hom T2437C genotypes were analyzed by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The haplotypes were calculated using PHASE 2.0 software.</p><p><b>RESULTS</b>VC%, FVC%, MVV%, FEV(1.0%) in exposed group were lower than in non-exposure group, the difference were significantly (P < 0.05). VC%, FVC%, MVV%, FEV1.0% in exposed group with HSP70-1, HSP70-2, HSP70-hom genotypes were lower than in non-exposure group (P < 0.05); FVC% in exposed group with HSP70-hom T/C genotypes were lower than that with HSP70-hom T/T genotypes, MVV% were lower than that with HSP70-hom T/T, C/C genotypes. There's no difference in pulmonary dysfunction index of HSP70-1, HSP70-2 genotypes (P>0.05), but significant difference between the exposed group with HSP70-1, HSP70-hom genotypes; The adjust OR (95%CI) of exposed group with HSP70-1 G/C genotypes and HSP70-homT/C genotypes were 2.516 (1.012 ∼6.252) and 2.284 (1.033∼5.053). Exposed group with CGT haplotype pulmonary dysfunction were significantly higher than in non-exposure group (P < 0.05).</p><p><b>CONCLUSION</b>Coke oven exposure may increase pulmonary dysfunction injury, Coke oven workers who have the HSP70-1 G/C genotypes, HSP70-hom T/C genotypes and CGT haplotypes may increase the susceptibility of pulmonary dysfunction. There must be some relationship between HSP70-1, HSP70-hom gene polymorphisms and lung function injury of Cock-oven Workers.</p>

Quantitative assessment of pulmonary function using low dose multi-slice spiral CT in smoker / 中华放射学杂志

ObjectiveTo evaluate the clinical feasibility of low dose MSCT for quantitative assessment of pulmonary function in smokers.MethodsOne hundred and forty-six patients with chronic objective pulmonary disease ( COPD ) including 109 smokers ( 74.6％ ) and 37 non-smokers ( 25.3％ )underwent pulmonary function test and low-dose MSCT scan.All data were analyzed using computer-aided lung anlysis software.Pulmonary function parameters from low-dose MSCT were compared between smokers and non-smokers and also compared with pulmonary function test in non-smokers ( Pearson test).Results In smokers,the average volume at full inspiratory phase (Vin) was (5125 ± 862 ) ml,mean lung attenuation was ( - 902 ± 26 ) HU,mean lung density was (0.0984 ± 0.0260 ) g/cm3,emphysema volume was (2890 ±1370) ml.The average volume at full expiratory phase (Vex) was (2756 ±1027) ml,mean lung attenuation was ( -811 ±62) HU,mean lung density was (0.1878 ±0.0631 ) g/cm3,emphysema volume was (685 ±104) ml.In non-smokers,the average Vin was (3734 ± 759) ml,mean lung attenuation was ( -876 ±40) HU,mean lung density was (0.1244 ±0.0401 )g/cm3,emphysema volume was ( 1503 ± 1217) ml.The average Vex was ( 1770 ± 679 ) ml,mean lung attenuation was ( - 765 ± 56 ) HU,mean lung density was (0.2360 ± 0.0563 ) g/cm3,emphysema volume was ( 156 ± 45 ) ml.There were significant differences between smokers and non-smokers (P ＜0.01 ).The Vex/Vin was correlated with residual volume/total lung capacity ( RV/TLC,r =0.60,P＜0.01 ),and Vin was correlated with TLC ( r =0.58,P ＜ 0.01 ),Vex with RV ( r =0.59,P＜0.01 ).Pixel index ( PI ) -950in was correlated with FEV 1％ pre and FEV1/FVC％ ( r =- 0.53,- 0.62,respective,P ＜ 0.01 ),Pl-950ex was correlated with FEV1 ％ pre and FEV1/FVC％ ( r =-0.71,-0.77,respective,P＜0.0l).ConclusionLow-dose MSCT can be a potential imaging tool for quantitative pulmonary function assessment in smokes.

Parameters of quantitative multi-slice CT imaging technology in pulmonary / 中国医学影像技术

Objective To evaluate the relevance of MSCT pulmonary function parameters and pulmonary function test (PFT) parameters, and to define the reference value of MSCT pulmonary function parameters. Methods Thirty male volunteers received clinical PFT and MSCT scan. MSCT scan was perfomed at the end of the maximum inspiratory and maximum expiratory. All data were analyzed with the lung analysis software of computer-aided inspection system correlatedly with pulmonary function parameters. Results The lung volume at full inspiratory volume (Vin) and full expiratory volume (Vex) in MSCT scan had good correlation with total lung capacity (TLC) and residual volume (RV) (r=0.90, P<0.01; r=0.74, P<0.01). Vex/Vin was correlated with RV/TLC (r=0.74, P<0.01), and Vin-Vex was correlated with MVC (r=0.85, P<0.01). In inspiration, the average lung density was (-879.51±32.82) HU, the density per unit volume was (0.12±0.03) g/cm3, while in expiratory they were (-688.14±62.38) HU and (0.31±0.06) g/cm3. Conclusion MSCT pulmonary function tests with the analysis software of computer-aided inspection system have good correlation with PFT.

Current status of prevention and management of chronic obstructive pulmonary disease in rural area in China / 中华内科杂志

Objective To investigate the current status of prevalence, prevention and management of chronic obstructive pulmonary disease(COPD) in rural area in China. Methods A cross-sectional survey of COPD was conducted in Beijing city, Shanghai city, Guangdong province, Liaoning province,Tianjin city, Chongqing province and Shanxi province. A population-based cluster sample was randomly selected from each rural area. In the selected community,all residents at least 40 years old were recruited,and interviewed with a modified standardized questionnaire from the international burden of obstructive lung diseases (BOLD) study. All participants were tested with spirometry. Those with airflow limitation were performed on post-bronchedilator spirometry. The post-bronchedilator a ratio of forced expiratory volume in one second to forced vital capacity (FEV1/FVC) less than 70% was defined as the diagnostic criteria of COPD. Results (1) Data of 9434 participants was valid for analysis, with a valid response rate of 83. 6% ;the prevalence of COPD in rural was 8. 8% (830/9434), 12. 8% in male and 5.4% in female. (2)The percentage of smoking and the exposure to biomass smoke in rural was 43. 0% (4059/9434) and 83. 1% (7835/9434) respectively; cigarettes cessation rate was 17.5% ; only 12. 4% ( 502/4059 ) of smokers had received advice to quit smoking. (3) Among COPD patients, only 30. 0% (249/830) had ever been diagnosed as COPD, bronchitis, emphysema, or asthma, 2. 4% (20/830) had ever received spirometric tests, and 74. 5% were current smokers; only 7.9% (50/634)COPD patients in stage two or over had received regular drug treatment. Conclusion There was high prevalence and poor prevention and management for COPD in rural areas. Therefore, an enforced prevention and management for COPD are urgent.

Implication of impulse oscillometry in the measurement of bronchial hyperresponsiveness / 中国医师进修杂志

0.05) in the negative group. However, there was a significant increase in each parameter except Rc in the positive group (P

Expression in E.coli JM109 and pharmacology on anti-angiogenesis of endogenetic angiogenesis inhibitor Arresten gene / 中国药理学通报

Aim To construct prokaryotic expression vector for endogenetic angiogenesis inhibitor Arresten gene and to express recombinant.Pharmacology experiments found that the expression product had the function of restrain blood vessel growing on the chorioallantoic membrane(CAM).Methods The total RNA was extracted from the placenta organize of the normal puerpera,and Arresten gene was amplified by RT-PCR to construct recombinant vector pBV220-Arr.Then,it was transformed into E.coli JM109 for yielding recombinant human Arresten protein.The method to detect inhibiting angiogenesis activity was CAM study.Results The plasmid pBV220-Arr.could be expressed in E.coli JM109 from two hours to eight hours,and the expression yield reached the highest in two hours.The Arresten protein could restrain blood vessel growth of CAM evidently than that of angiostatin.Conclusion The recombinant plasmid pBV220-Arr was construted successfully and the target protein Arresten can express in E.coli JM109,the protein of Arresten had the evident effect of inhibite the activity of angiogenesis.

Normative values of pulmonary function testing in Chinese adults / 中华医学杂志(英文版)

<p><b>OBJECTIVES</b>To compare the difference in pulmonary function between Caucasians and Chinese and assess the best reference values of pulmonary function for Chinese adults.</p><p><b>METHODS</b>Values for pulmonary function were predicted by tests on 4773 Chinese healthy subjects (male: 2560, female: 2213, aged 15-78 years) in six parts (north, northeast, northwest, east, southwest and south) of China. Prediction equations of the European Community for Steel and Coal (ECSC), other equations for overseas Chinese or for Caucasians were also selected. The regression coefficients of forced vital capacity (FVC), forced expiratory volume in one second (FEV1), FEV1/FVC, total lung capacity (TLC) and residual volume (RV) were summarized.</p><p><b>RESULTS</b>ECSC predictions were closer to the Chinese ones than other selected equations. Comparison with ECSC predictions showed that on average the values for FVC, FEV1 were 5.3% smaller in Chinese males and 3.3% smaller in Chinese females, with the maximal differences in south China and the minimal differences in North China. RV and TLC in Chinese were lower than in Caucasians (males 4.8%, 5.5%, respectively; females 8.7% and 6.0%, respectively). Conversion factors were given for adjusting ECSC equations to fit Chinese.</p><p><b>CONCLUSIONS</b>For predicting values of pulmonary function in Chinese, we suggest to use the equations reported here. Alternatively, ECSC regression equations may be used with appropriate conversion factors.</p>

The Assessment of Bronchial hyperresponsiveness in 187 elderly subjects / 中华老年医学杂志

0.05). The BHR distribution was slight 31.9%, mild 43.5%, moderate 24.6%, no severe BHR were found in this study, showing less correlation between the percentage of predicted FEV 1 and the PD 20FEV 1-His in the elderly (r=0.277, P

The application of phrenic nerve conduction time in the resuscitation of anaesthesia / 中国病理生理杂志

AIM: To investigate the application of phrenic nerve conduction time(PNCT) in the recovery of general anaesthesia. METHODS: Pdi (t) and PNCT were measured by cervical magnetic stimulation of phrenic nerves in 8 patients before and after the administration of muscle relaxant. RESULTS: Pdi (t) was reduced from (23.7?2.4) cmH 2O to (11.5?3.4) cmH 2O (reduction rate: 51.5%, P