[Value of peripheral blood rare cell EGFR gene amplification detection in the evaluation of benign and malignant pulmonary nodules].

Objective: To explore the value of detection of epidermal growth factor receptor (EGFR) gene amplification in peripheral blood rare cells in the assessment of benign and malignant pulmonary nodules. Methods: A total of 262 patients with pulmonary nodules were selected as the retrospectively study subjects from the Second Affiliated Hospital of Army Military Medical University and Peking Union Medical College Hospital from July 2022 to August 2023. There were 98 males and 164 females, with the age range from 16 to 79 (52.1±12.1) years. The EGFR gene amplification testing was performed on the rare cells enriched from patients' peripheral blood, and the clinical manifestations, CT imaging features, histopathological and/or pathological cytological confirmed results of patients were collected. The receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value of the method of detection of EGFR gene amplification in peripheral blood rare cells, and its diagnostic efficacy was evaluated. Results: Among the 262 patients, 143 were malignant pulmonary nodules and 119 were benign pulmonary nodules. The differences between malignant pulmonary nodules and benign pulmonary nodules in nodule diameter and nodule density were statistically significant (both P<0.001), while the differences in age, gender and nodule number were not statistically significant (all P>0.05). The number [M (Q1, Q3)] of EGFR gene amplification positive rare cells in patients with malignant pulmonary nodule was 8 (6, 11), which was higher than that in patients with benign pulmonary nodule [2 (1, 4), P<0.001]. The ROC curve results showed that when the optimal cut-off value was 5 (that was, the number of EGFR gene amplification positive rare cells was>5), the area under the curve (AUC) of the detection of EGFR gene amplification in peripheral blood rare cells for discrimination of benign and malignant pulmonary lesions was 0.816 (95%CI: 0.761-0.870), with a sensitivity of 83.2%, a specificity of 80.7%, and an accuracy of 82.1%. Based on the analysis of the diameter of the nodules, the AUC for distinguishing between benign and malignant pulmonary nodules with diameter 5-9 mm and 10-30 mm was 0.797 (95%CI: 0.707-0.887) and 0.809 (95%CI: 0.669-0.949), respectively, with sensitivity, specificity and accuracy reached 75% or above. Based on the analysis of nodule density, the AUC for distinguishing between benign and malignant solid nodule and subsolid nodule was 0.845 (95%CI: 0.751-0.939) and 0.790 (95%CI: 0.701-0.880), respectively, with sensitivity, specificity and accuracy reached 75% or above. Based on the analysis of nodule number, the AUC for distinguishing between benign and malignant solitary pulmonary nodule and multiple pulmonary nodule was 0.830 (95%CI: 0.696-0.965) and 0.817 (95%CI: 0.758-0.877), respectively, with sensitivity, specificity and accuracy reached 80% or above. Conclusion: The detection of EGFR gene amplification in peripheral blood rare cells contributes to the evaluation of benign and malignant pulmonary nodules, and can be used in the auxiliary diagnosis of benign and malignant pulmonary nodules.

[The expression and function of PD-L1 in CD133(+) human liver cancer stem-like cells].

Objective: To investigate the expression of programmed death protein-ligand 1 (PD-L1) in liver cancer stem-like cells (LCSLC) and its effect on the characteristics of tumor stem cells and tumor biological function, to explore the upstream signaling pathway regulating PD-L1 expression in LCSLC and the downstream molecular mechanism of PD-L1 regulating stem cell characteristics, also tumor biological functions. Methods: HepG2 was cultured by sphere-formating method to obtain LCSLC. The expressions of CD133 and other stemness markers were detected by flow cytometry, western blot and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the expressions of stemness markers and PD-L1. The biological functions of the LCSLC were tested by cell function assays, to confirm that the LCSLC has the characteristics of tumor stem cells. LCSLC was treated with cell signaling pathway inhibitors to identify relevant upstream signaling pathways mediating PD-L1 expression changes. The expression of PD-L1 in LCSLC was down regulated by small interfering RNA (siRNA), the expression of stem cell markers, tumor biological functions of LCSLC, and the changes of cell signaling pathways were detected. Results: Compared with HepG2 cells, the expression rate of CD133 in LCSLC was upregulated [(92.78±6.91)% and (1.40±1.77)%, P<0.001], the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were also higher than those in HepG2 cells (P<0.05), the number of sphere-formating cells increased on day 7 [(395.30±54.05) and (124.70±19.30), P=0.001], cell migration rate increased [(35.41±6.78)% and (10.89±4.34)%, P=0.006], the number of transmembrane cells increased [(75.77±10.85) and (20.00±7.94), P=0.002], the number of cloned cells increased [(120.00±29.51) and (62.67±16.77), P=0.043]. Cell cycle experiments showed that LCSLC had significantly more cells in the G(0)/G(1) phase than those in HepG2 [(54.89±3.27) and (32.36±1.50), P<0.001]. The tumor formation experiment of mice showed that the weight of transplanted tumor in LCSLC group was (1.32±0.17)g, the volume is (1 779.0±200.2) mm(3), were higher than those of HepG2 cell [(0.31±0.06)g and (645.6±154.9)mm(3), P<0.001]. The expression level of PD-L1 protein in LCSLC was 1.88±0.52 and mRNA expression level was 2.53±0.62, both of which were higher than those of HepG2 cells (P<0.05). The expression levels of phosphorylation signal transduction and transcription activation factor 3 (p-STAT3) and p-Akt in LCSLC were higher than those in HepG2 cells (P<0.05). After the expression of p-STAT3 and p-Akt was down-regulated by inhibitor treatment, the expression of PD-L1 was also down-regulated (P<0.05). In contrast, the expression level of phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in LCSLC was lower than that in HepG2 cells (P<0.01), there was no significant change in PD-L1 expression after down-regulated by inhibitor treatment (P>0.05). After the expression of PD-L1 was knockdown by siRNA, the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were decreased compared with those of siRNA-negative control (NC) group (P<0.05). The number of sphere-formating cells decreased [(45.33±12.01) and (282.00±29.21), P<0.001], the cell migration rate was lower than that in siRNA-NC group [(20.86±2.74)% and (46.73±15.43)%, P=0.046], the number of transmembrane cells decreased [(39.67±1.53) and (102.70±11.59), P=0.001], the number of cloned cells decreased [(57.67±14.57) and (120.70±15.04), P=0.007], the number of cells in G(0)/G(1) phase decreased [(37.68±2.51) and (57.27±0.92), P<0.001], the number of cells in S phase was more than that in siRNA-NC group [(30.78±0.52) and (15.52±0.83), P<0.001]. Tumor formation in mice showed that the tumor weight of shRNA-PD-L1 group was (0.47±0.12)g, the volume is (761.3±221.4)mm(3), were lower than those of shRNA-NC group [(1.57±0.45)g and (1 829.0±218.3)mm(3), P<0.001]. Meanwhile, the expression levels of p-STAT3 and p-Akt in siRNA-PD-L1 group were decreased (P<0.05), while the expression levels of p-ERK1/2 and ß-catenin did not change significantly (P>0.05). Conclusion: Elevated PD-L1 expression in CD133(+) LCSLC is crucial to maintain stemness and promotes the tumor biological function of LCSLC.

[Profiles and differences of submucosal microbial in peri-implantitis and health implants: A cross-sectional study].

OBJECTIVE: To describe the submucosal microbial profiles of peri-implantitis and healthy implants, and to explore bacteria that might be correlated with clinical parameters. METHODS: In the present cross-sectional study, 49 patients were recruited. Each patient contributed with one implant, submucosal biofilms were collected from 20 healthy implants and 29 implants with peri-implantitis. DNA was extracted and bacterial 16S ribosomal RNA (16S rRNA) genes were amplified. Submucosal biofilms were analyzed using 16S rRNA sequencing at Illumina MiSeq platform. Differences between the groups were determined by analyzing α diversity, microbial component and microbial structure. The potential correlation between the bacteria with pocket probing depth (PPD) of peri-implant calculated by Spearman correlation analysis. RESULTS: The α diversity of submucosal microbial of health group was significantly lower than that in peri-implantitis group (Chao1 index: 236.85±66.13 vs. 150.54±57.43, P < 0.001; Shannon index: 3.42±0.48 vs. 3.02±0.65, P=0.032). Principal coordinated analysis showed that the submucosal microbial structure had significant difference between healthy and peri-implantitis groups [R2=0.243, P=0.001, analysis of similarities (ANOSIM)]. Compared with healthy implants, relative abundance of periodontal pathogens were higher in peri-implantitis, including members of the red complex (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and some members of orange complex (Precotella intermedia, Eubacterium nodatum, Parvimonas micra), as well as some new periodontal pathogens, such as Fillifactor alocis, Fretibacterium fastidiosum, Desulfobulbus sp._HMT_041, and Porphyromonas endodontalis. Spearman correlation analysis revealed that the relative abundance of Treponema denticola (r=0.686, P < 0.001), Tannerella forsythia (r=0.675, P < 0.001), Fretibacterium sp. (r=0.671, P < 0.001), Desulfobulbus sp._HMT_041 (r=0.664, P < 0.001), Filifactor alocis (r=0.642, P < 0.001), Fretibacterium fastidiosum (r=0.604, P < 0.001), Porphyromonas gingivalis (r=0.597, P < 0.001), Porphyromonas endodontalis (r=0.573, P < 0.001) were positive correlated with PPD. While the relative abundance of Rothia aeria (r=-0.615, P < 0.001) showed negatively correlation with PPD. CONCLUSION: Marked differences were observed in the microbial profiles of healthy implants and peri-implantitis. The members of red and orange complex as well as some new periodontal pathogens seem to play an important role in peri-implant disease. Compared with healthy implants, the submucosal microbial of peri-implantitis were characterized by high species richness and diversity.

[Correlation between gene polymorphisms of killer cell immunoglobulin-like receptors and their ligands and Graves' disease].

Objective: To explore the relationship between gene polymorphism of killer cell immunoglobulin-like receptor (KIR) and its ligand-specific human leukocyte antigen C (HLA-C) and Graves' disease (GD). Methods: Case-control study. A total of 118 unrelated GD patients (GD group) admitted to Shandong Provincial Hospital from January 2011 to December 2017 and 108 age-and sex-matched healthy controls (healthy control group) were included. The KIR genotype and its ligand HLA-C allele were detected by polymerase chain reaction sequence-specific primers (PCR-SSP). The distribution of KIR/HLA-C gene combination in GD patients and control population was analyzed to explore its association with the occurrence of GD. Results: In GD group, there were 29 males and 89 females, aged (38±14) years. In the healthy control group, there were 28 males and 80 females, aged (37±13) years. Compared with the healthy control group, the occurrence frequency of HLA-Cw01 was higher in GD group[36.4%(43/118) vs 18.5%(20/108), P=0.003], and the occurrence frequency of HLA-Cw03 and HLA-Cw06 was lower in GD group[11.9%(14/118) vs 39.8%(43/108), P<0.001; 9.3%(11/118) vs 18.5%(20/108), P=0.045]. The frequency of KIR2DL1/HLA-C2 gene combination in GD group was lower than that in control group [17.8%(21/118) vs 34.3%(37/108), P=0.005]. Logistic regression analysis showed that KIR2DL1/HLA-C2 gene combination was a protective factor for GD occurrence (OR=0.308, 95%CI: 0.126-0.752, P=0.010). Conclusions: The polymorphism of KIR/HLA-C gene is related to GD. The low expression of KIR2DL1/HLA-C2 in GD patients may be a protective factor for GD.

[Association between exposure patterns of adverse childhood experiences and anxiety symptom trajectories in medical college students].

Objective: To analyze the association between exposure patterns of adverse childhood experiences (ACEs) and anxiety symptom trajectories in medical college students. Methods: A survey was conducted on first-year students from Anhui Medical College and Anqing Medical College, using the Childhood Abuse Questionnaire, Family Disability Questionnaire, Childhood Adverse Social Experience Item, and Anxiety Self Rating Scale. The baseline survey was conducted from November to December 2019, and two follow-up visits were conducted once every six months until November to December 2020. The latent class analysis (LCA) was used to analyze the exposure patterns of ACEs. The latent class growth analysis (LCGA) was used to analyze the development trajectory of anxiety symptoms. The multiple logistic regression model was used to analyze the correlation between different exposure patterns of ACEs and the trajectory of anxiety symptom trajectories. Results: A total of 3 662 college students aged (19.2±1.0) were surveyed. The LCA showed that the exposure patterns of ACEs could be divided into the "high ACEs" group (13.4%), "high neglect/emotional abuse" group (25.7%), "high family dysfunction" group (6.9%), "high neglect" group (27.1%), and "low ACEs" group (26.3%). The LCGA divided anxiety trajectories into four groups: "high anxiety decline" (7.1%),"anxiety increase "(4.1%), "moderate anxiety"(52.9%), and "low anxiety"(35.9%). Using the low ACEs group as a reference group, compared with the low anxiety trajectory, the high ACEs group, high neglect/emotional abuse group, high family dysfunction group, high neglect group, and medium to high-level anxiety trajectory were all associated with an increased risk (P<0.05). Conclusion: There is heterogeneity in ACEs exposure patterns among medical college students, and ACEs exposure patterns are important influencing factors for anxiety symptom trajectories.

[Application of artificial intelligence in digital chest radiography diagnosis of pneumoconiosis].

Pneumoconiosis is the occupational disease with the highest burden in China currently. The diagnosis of pneumoconiosis mainly relies on manual reading of X-ray high-kilovoltage or digital photography chest radiograph, which has some problems such as low efficiency, strong subjectivity, and cannot accurately judge the critical lesions. With the progress of machine-aided diagnosis technology, the efficient, objective and quantitative of artificial intelligence diagnosis technology just solve the shortcomings above. This paper reviews the research progress in digital chest radiography diagnosis of pneumoconiosis using artificial intelligence technology, especially deep learning model, combined with the limitations of conventional manual reading, in order to clarify the application prospect of artificial intelligence technology in the diagnosis of pneumoconiosis by digital chest radiography, and provide a direction for future research in this field.

[Analysis on the quality control of suspected occupational disease from the characteristics of applicants diagnosed with noise deafness].

Objective: To analyze the audiology and occupational health data of applicants diagnosed of occupational noise deafness, and to explore the influencing factors in the diagnosis of suspected occupational noise deafness. Methods: In May 2022, the information of patients diagnosed with occupational noise deafness in Peking University Third Hospital from January 2018 to December 2021 was collected, and the occupational health data of their working environment, clinical audiological examination results and diagnosis basis of occupational noise deafness were collected and analyzed. Multi-factor unconditional logistic regression analysis was used to analyze independent risk factors for the diagnosis of occupational noise deafness. Results: A total of 129 subjects were included, all of which were suspected cases of occupational noise deafness found in various occupational health examination institutions. Eight cases (6.20%) were diagnosed as occupational noise deafness, and 121 cases (93.80%) were non-occupational noise deafness. After hearing examination, only 27.27% (24/88) of the patients' audiological changes were consistent with the starting point of occupational noise deafness diagnosis. Further analysis of the noise intensity in the workplace showed that 16 patients were identified as non-occupational noise deafness because the noise intensity of the working environment was less than 85 dB. Logistic regression analysis showed that the working hours were more than 8 hours (OR=9.274, 95%CI: 1.388-61.950, P=0.022) and the noise intensity of the working environment (OR=1.189, 95%CI: 1.059-1.334, P=0.003) were independent risk factors for the diagnosis of occupational noise deafness. Conclusion: The exclusion rate of suspected occupational noise deafness found in occupational health examination is higher after adequate rest. The test results of working environment noise intensity provided by the employer can help to determine occupational noise deafness.

[Efficacy of combined application of glycine powder air-polishing in non-surgical treatment of peri-implant diseases].

OBJECTIVE: To compare the clinical efficacy of combined application of glycine powder air-polishing and mechanical submucosal debridement in non-surgical treatment of peri-implant diseases. METHODS: A randomized controlled clinical study was carried out on patients diagnosed with peri-implant diseases in the Department of Periodontology, Peking University School and Hospital of Stomatology, between May of 2020 and June of 2021.Twenty-eight patients with totally sixty-two implants were enrolled.The patients were randomly divided into the test group and control group. The patients in the test group (13 subjects/32 implants) received mechanical submucosal debridement using titanium curettes combined with application of glycine powder air-polishing, while the control group (15 subjects/30 implants) received mechanical submucosal debridement using titanium only. Clinical parameters, such as plaque index (PLI), pocket probing depth (PPD), bleeding index (BI) and the percentage of suppuration on probing on implants' level (SoP%) were measured at baseline and 8 weeks after non-surgical intervention. Changes and group differences of clinical parameters of the implants before and 8 weeks after non-surgical intervention were compared. RESULTS: Mean PLI, PPD, BI of both the test group and control group significantly reduced 8 weeks after non-surgical intervention (P < 0.05). Compared with the control group, the test group achieved lower BI (2.7±0.8 vs. 2.2±0.7, P < 0.05), more reduction of BI (0.6±0.7 vs. 1.1±0.6, P < 0.01) and more reduction of SoP% (21.9% vs. 10%, P < 0.05) after non-surgical intervention. Both the control and test groups exhibited comparable PLI and PPD reductions (P>0.05). For the implants diagnosed with peri-implant mucositis, the test group revealed more signi-ficant reduction in BI and SoP% than the control group (1.0±0.7 vs. 0.4±0.7, P=0.02; 6.3% vs. 0, P=0.012). There was no significant difference existing in PLI and PD improvement between the control group and test group (P>0.05). For the implants diagnosed with peri-implantitis, there was no significant difference existing in PLI, PPD, BI and SoP% improvement values between the test and control groups (P>0.05). No complications or discomforts were reported during the study. CONCLUSION: Both treatment procedures could relieve the inflammation of peri-implant soft tissue. Non-surgical mechanical submucosal debridement combined application of glycine powder air-polishing is associated with significant reduction of soft tissue bleeding and suppuration on probing especially in the implants diagnosed with peri-implant mucositis.

[Diagnostic value of surgical lung biopsies for diffuse parenchymal lung disease: the change of disease spectrum in the past 28 years in a single institution in China].

Objective: To investigate the changes of disease spectrum in diffuse parenchymal lung disease (DPLD) diagnosed by surgical lung biopsy, and to explore the diagnostic value of surgical lung biopsy in DPLD. Methods: Four hundred and fifty-five consecutive DPLD patients, who underwent surgical lung biopsy in Peking Union Medical College Hospital during the past 28 years, were analyzed retrospectively. Results: There were 211 males and 244 females. The average age at biopsy was (45±14) years. Four hundred and eleven cases (90.3%) were diagnosed by pathologic findings. Four hundred and forty-one cases (96.9%) were diagnosed by clinical-radiologic-pathologic multidisciplinary discussion. The 30-day mortality and 90-day mortality were 2.4% and 3.3% respectively. The disease spectrum included interstitial pneumonia in 209 cases (45.9%) (nonspecific interstitial pneumonia in 105 cases, usual interstitial pneumonia in 33 cases), other miscellaneous DPLD in 166 cases (36.5%) (including hypersensitivity pneumonitis in 49 cases), tumor in 39 cases (8.6%), and infectious diseases in 27 cases (5.9%). In the three consecutive periods (1993-2002, 2003-2012 and 2013-2020), the number of biopsies was 76 (16.7%), 297 (65.3%) and 82 (18%) respectively. The disease spectrum changes over time: in the above three periods, the percentage of interstitial pneumonia in DPLD was 68.4%, 45.1% and 28%, other miscellaneous DPLDs were 22.4%, 39.4% and 39.0%, the tumors were 2.6%, 7.4% and 18.3%, the infectious diseases were 5.3%, 5.1% and 9.8%. Conclusions: This study presented the changes of disease spectrum in DPLD diagnosed by surgical lung biopsy through single center real-world data, reflecting the progress of clinicians' understanding of DPLD and interstitial pneumonia. Surgical lung biopsy is still valuable for some difficult and complicated DPLD cases.

High cyclic tensile stress disrupts the extracellular matrix in human chondrocyte by F-actin cytoskeletal polymerization and reactive oxygen species production.

This study aims to explore the mechanism of cyclic tensile stress (CTS) on human chondrocytes (CHs) relating to the reactive oxygen species (ROS) generation and extracellular matrix (ECM) stability in vitro. A well-established CTS model with 5%, 10%, or 20% elongation was performed for CHs stretching. After CTS, the cell viability, total ROS level, main ECM components, matrix metalloproteinase (MMP), tissue inhibitor of metalloproteinase (TIMP), F-actin density, and some anti-oxidative enzymes were analyzed. Additionally, the antioxidant N-acetylcysteine (NAC) and cytochalasin D were used to suppress the ROS production and F-actin polymerization when the CHs underwent CTS, respectively. The treatment of 20% elongation-CST significantly decreased the CH viability and the expressions of collagen II, aggrecan, anti-oxidative enzymes and TIMP3/4, however, it increased the ROS accumulation, F-actin polymerization, and the expression of collagen I and MMP3/13. In contrast, the application of NAC and cytochalasin D could partly rescue the CHs from the injury caused by the high CTS. Therefore, high CTS disrupts the ECM by remodeling the F-actin cytoskeleton and promoting ROS production. Cytochalasin D and NAC are effective in rejecting F-actin cytoskeleton polymerization, and ROS accumulation through a potential synergetic process, which alleviates the ECM injury caused by High CTS.

[Efficacy of high-voltage long-duration pulsed radiofrequency treatment in patients with neuralgia resulting from failed back surgery syndrome].

Objective: To explore the efficacy and safety of high-voltage long-duration pulsed radiofrequency (PRF) treatment in patients with neuralgia resulting from failed back surgery syndrome (FBSS). Methods: The clinical data of 58 patients diagnosed with neuralgia resulting from FBSS in the Department of Pain Medicine, Peking University Third Hospital from January 2017 to January 2020 were retrospectively analyzed. The patients were divided into two groups according to the treatment method. Experimental group (n=28) underwent high-voltage long-duration PRF therapy, using ultrasound and X-ray guidance to target the spinal nerve of the affected side, while control group (n=30) was applied with the standard pulsed radiofrequency therapy. Visual analogue scale (VAS), Oswestry disability index (ODI), 36-item short form health survey (SF-36), patient health questionnaire (PHQ-9) before treatment and at 1 week, 1 month, and 6 months after treatment were recorded. Meanwhile, postprocedural complications and adverse reactions were also collected. Results: VAS, ODI, SF-36 and PHQ-9 scores at 1 week, 1 month, and 6 months after treatment were significantly improved in both groups compared with their respective pre-treatment baseline scores (all P<0.01). The differences of VAS, ODI, and PHQ-9 scores between the two groups were not statistically significant at 1 month after treatment (all P>0.05). However, VAS, ODI, and PHQ-9 scores were lower in experiment group than those in control group at 6 months after treatment (all P<0.05). The marked improvement rate and total effective rate at 6 months after treatment in experiment group was 78.6% (22/28) and 92.9% (26/28), respectively, which were higher than that of control group [60.0% (18/30) and 83.3% (25/30), respectively], but the differences were not statistically significant (both P>0.05). No serious complications occurred during the whole period of treatment. Conclusions: Both treatments can effectively relieve the lower limb neuralgia. High-voltage long-term PRF has better efficacy and longer duration than standard PRF.

[Effects of acute phosgene exposure on kidney in rats].

Objective: To investigate the changes in kidney and its mechanism during the development of acute phosgene exposure in rats. Methods: Rats were randomized into 2 groups: control and phosgene group (including 1, 3, 6, 12 and 24 h after exposed to phosgene) , 6 rats in each group. Rats in control group were exposed to air for 5 min, while rats in phosgene group were exposed to 8.33 mg/L phosgene for 5 min. The blood samples were collected at 1, 3, 6, 12 and 24 h after phosgene exposure. The blood creatinine (Cr) , urea nitrogen (BUN) and blood gas analysis were detected. HE staining and immunohistochemical staining were performed to observe the expression levels of 8-hydroxy deoxyguanosine and myeloperoxidase. Results: The arterial partial pressure of oxygen and oxygenation index of rats in the phosgene group were significantly lower than those in the control group at 3, 6 and 12 h after exposure (P<0.01) . The lowest points were reached at 6 h, which were (58.67±7.89) mmHg and (202.30±27.20) mmHg, respectively. The Cr and BUN of rats in the phosgene group were significantly higher than those in the control group at 3, 6, 12, and 24 h, and the renal organ coefficients were significantly higher than those in the control group at 3, 6 and 12 h (P<0.01) . HE staining showed that there were more erythrocytes in the glomeruli of rats in the phosgene group, the volume of renal tubular epithelial cells increased, and the cytoplasm was loose and lightly stained. The damage was most obvious at 6 h. The results of immunohistochemical staining showed that the positive expressions of 8-hydroxy deoxyguanosine and myeloperoxidase in the kidney tissue of the phosgene group increased. Conclusion: Hypoxemia and oxidative stress caused by phosgene poisoning may be the cause of renal damage in rats.

[Analysis of anxiety, depression and related factors in patients with chronic lumbocrural pain before minimally invasive surgery].

OBJECTIVE: To investigate anxiety and/or depression status of patients with chronic lumbocrural pain, and to further analyze related risk factors of anxiety and/or depression . METHODS: Retrospective analysis of the medical data of patients who suffered from chronic lumbocrural pain caused by lumbar disc herniation and/or lumbar spinal stenosis and received minimally invasive surgery from March 2018 to April 2018. General data (including age, gender, education levels, past history, sleep order and medical insurance), numeric rating scale(NRS), Japanese Orthopedic Association(JOA) back pain scale and hospital anxiety and depression scale(HADS) were collected for analysis. The basic demographic data and clinic data were analyzed, possible related risk factors associated were analyzed by univariate analysis, and multivariate Logistic regression analysis was further used to find the relative independent risk factors and included all the predictive variables with P values less than 0.05 as covariates. RESULTS: A total of 91 patients met the inclusion criteria and finished this study, the mean HADS score for anxiety was 8.1±4.2, 48(52.7%) respondents were screened positive for anxiety, while the rest 43(47.3%) patients had negative anxiety state, the mean HDDS score for depression was 6.9±4.9, 38(41.8%) respondents were screened positive for depression, and the rest 53(58.2%) patients were not depressed, and 56(61.5%) patients experienced anxiety or depression. There were significant difference in sleep disorder, JOA score and leg NRS score between the patients with and without anxiety(P<0.05), and the significant differences were also found in age, sleep disorder and JOA score between the patients with and without depression(P<0.05), Logistic regression analysis further showed that the JOA score and sleep disorder were risk factors for anxiety, and the JOA score was risk factor for depression. CONCLUSION: Patients with chronic lumbocrural pain are often accompanied by anxiety and/or depression before minimally surgery, the low JOA score and sleep disturbance increased the risk of presenting anxiety, and the low JOA score increased the risk of developing depression. It is necessary to evaluate mental status and related risk factors before surgery.

[Analysis on characteristic of patients with stage I coal worker's pneumoconiosis].

Objective: To compare the clinical and imaging characteristics of patients with stage I coal worker's pneumoconiosis (CWP) . Methods: All 347 cases of coal workers' pneumoconiosis diagnosed in the Third Hospital of Peking University from January 2014 to December 2018 were included in the study. According to different working posts, the subjects were divided into three categories: mining, tunneling and mixing workers. Dust exposure duration, initial dust exposure age, diagnosis age, latency, small shadow shape and lung regions distribution in X-ray chest film of different categories of CWP patients were analyzed. Results: Among the 347 patients, 216 were mining workers (62.2%) , 77 were tunneling workers (22.2%) and 54 were mixing workers (15.6%) . The dust exposure duration of mining, tunneling and mixing workers were (14.5±7.0) , (16.3±8.2) and (19.0±8.8) years, respectively. There are statistically significant differences in dust exposure duration between different categories of workers (P<0.05) . There were no significant difference in the age of diagnosis, initial dust exposure age and the latency between different categories of workers (P>0.05) . The X-ray films of mining, tunneling and mixing workers showed small round shadow, accounting for 50.9% (110/216) , 96.1% (74/77) and 96.3% (52/54) respectively. 48.1% (104/216) of the mining workers and 38.9% (21/54) of mixing workers, the distribution of small shadow in chest X-ray films reached middle and lower lung regions, while in the 48.1% (37/77) of the tunneling workers, the distribution of small shadow in chest X-ray films reached lower lung regions. There were differences in above indicators among workers with different categories (P<0.05) . Conclusion: The dust exposure duration, the shape and the distribution of lung area on chest X-ray films are different in stage I CWP patients of different occupational categories.

[Investigation of the relationship between occupational gasoline exposure and metabolic syndrome].

Objective: To explore the correlation between occupational gasoline exposure and metabolic syndrome (MS) . Methods: In September 2019, a total of 147 occupational gasoline exposure workers from a oil sales company in Beijing were selected as the observation group by using cluster sampling method, 158 people without gasoline exposure from the company were selected as the control group. Occupational health examination were performed to measure body mass, blood pressure and fasting plasma glucose (FPG) , triglycerides (TG) , high-density lipoprotein cholesterol (HDL-C) and other data. General demographic characteristics, occupational history, past medical history and personal history were analyzed either. Results: The levels of systolic blood pressure, diastolic blood pressure, FPG, TG and BMI in the observation group were all higher than those in the control group (P<0.05) . The detection rates of MS, obesity and hypertension in the observation group were all higher than those in the control group (P<0.05) . There were no significant differences in age, gender, working age, drinking, smoking, marital status, HDL-C level, detection rates of abnormal TG and HDL-C between the observation group and the control group (P>0.05) . The odds ratio (OR) of MS in the observation group was 1.988 times that in the control group (P<0.05) . Conclusion: Occupational gasoline exposure is associated with the increasing detection rate of MS.

[The study on the role of extracellular histones in the pathogenesis of coal worker's pneumoconiosis].

Objective: To explore the role of extracellular histones in the pathogenesis of coal worker's pneumoconiosis (CWP) , the relationship of extracellular histones in plasma with pulmonary fibrosis caused by coal mine dust was analyzed, and the stimulating effect of extracellular histones on fibroblast proliferation was studied. Methods: In May 2019, a total of 220 coal mine dust exposure workers (including coal miners and CWP patients) who visited the occupational disease outpatient department of Peking University Third Hospital from 2012 to 2015 were enrolled in the study. According to the classification of small opacity profusion (SOP) in chest radiograph for pneumoconiosis diagnosis (category 0, 1, 2, 3) , 61 coal miners were in category 0 SOP, 65 coal miners were in category 1 SOP, 56 coal miners were in category 2 SOP and 38 coal miners were in category 3 SOP. The plasma levels of extracellular histone H4 and platelet-derived growth factor (PDGF) were measured by enzyme-linked immunosorbent assay (ELISA) kit. The stimulating effects of CWP patients' plasma and calf thymus histones (CTHs) on fibroblast and the antagonizing effect of anti-H4 antibody were investigated by fibroblast proliferation experiment in vitro. Results: Among the study subjects, there were 195 males (88.6%, 195/220) and 25 females (11.4%, 25/220) , age (55.1±7.2) years, coal mine dust exposure time (16.3±4.4) years. The plasma concentrations of histone H4 in the coal miners with category 0, 1, 2 and 3 SOP were (3.92±1.75) 、(9.84±4.17) 、(14.35±5.52) and (17.83±7.69) µg/ml, respectively. There were significant differences among the four groups (P<0.01) . The plasma level of histone H4 was positively correlated with the plasma level of PDGF in the coal miners (r=0.769, P<0.01) . Compared with healthy control plasma group, the cell proliferation percentages of patients' plasma group (272%±87%) and CTH group (283%±84%) were significantly increased (P<0.05) . Compared with patients' plasma group, the cell proliferation percentage of patients' plasma+anti-H4 antibody group (185%±66%) was significantly decreased (P<0.05) . Compared with CTH group, the cell proliferation percentage of CTH+anti-H4 antibody group (167%±59%) was significantly decreased (P<0.05) . Conclusion: Extracellular histones in plasma are associated with pulmonary fibrosis in patients with CWP. Studies in vitro have shown that extracellular histones can promote proliferation of pulmonary fibroblasts. It is suggested that extracellular histones can be important biomarkers for pulmonary fibrosis caused by coal mine dust.

[Characteristics of initial diagnosis in autoimmune liver disease: an 18-year, retrospective single-center study].

Objective: To analyze the chorological changes of diagnosis in patients with autoimmune liver disease (AILD) and related factors for early diagnosis. Methods: A total of 581 patients with age ranged from 16 to 81 were retrospectively analyzed, who were admitted to Tianjin Medical University General Hospital with AILD during January 2000 to December 2017. Age at diagnosis, diagnostic method and cirrhosis at diagnosis were compared in different groups according to admission period as 2000-2005, 2006-2011, 2012-2017. Results: The diagnostic rate of AILD showed an upward trend during the past near two decades. The proportion of AILD patients diagnosed via health examination was increasing year by year mainly by elevated transaminases (P<0.001). The mean age at diagnosis in our AILD patients were younger at present, especially in men (P=0.044). The proportion of cirrhosis at diagnosis was gradually reduced in three different periods respectively [77.78%(21/27), 41.58% (79/190), 25.00%(91/364), P<0.001], which were coincident in patients with autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) (P<0.001). The shrinking trend of cirrhosis at diagnosis was significantly correlated with the increasing application of health examination (r=-0.549, P<0.001). Conclusions: Extensive application of health examination expands the diagnostic rate of AILD. During the past 18 years, more young patients are diagnosed with AILD. The proportion of severe cases such as cirrhosis at diagnosis is decreasing. Screening of immunological examinations in patients with abnormal transaminases is needed and critical to the early diagnosis of asymptomatic AILD.

[Activation of lung endothelial cells by extracellular histone in mice with acute respiratory distress syndrome].

Objective: To observe the changes of extracellular histones and pulmonary microvascular endothelial cells, and study the activating role of extracellular histones to pulmonary microvascular endothelial cells in the pathogenesis of acute respiratory distress syndrome (ARDS) . Methods: The correlation of the severity of acute lung injury with extracellular histones and pulmonary endothelial damage was studied through mice model, and acute lung injury was produced by aspiration of different concentrations of hydrochloric acid (0.01、0.1、0.3 and 0.5 mol/L, 2 ml/kg). Tumor necrosis factor-α (TNF-α), soluble thrombomodulin (sTM) and lung pathological change were measured. The pro-inflammatory role of extracellular histones was tested by injecting calf thymus histones (CTH) or specific anti-H4 antibody through tail vein. The direct activating role of extracellular histones to pulmonary microvascular endothelial cells was studied through pulmonary endothelial model. Results: The extracellular histones in plasma were increased obviously 6h after aspiration of different concentrations of hydrochloric acid in mice. A positive correlation was seen between extracellular histones and concentrations of aspirated hydrochloric acid (r=0.9180, P<0.05). The sTM in plasma also showed a positive correlation with concentrations of aspirated hydrochloric acid (r=0.8701, P<0.05). Merely administering CTH could not only increase TNF-α and sTM in plasma but also cause obvious lung injury, while specific anti-H4 antibody could relieve the inflammation and lung damage caused by CTH. Extracellular histones could directly damage pulmonary endothelial cells to release sTM in pulmonary endothelial model in vitro, while anti-H4 antibody could protect the endothelial cells. Conclusion: Extracellular histones are the key endogenic inflammatory mediators during the pathogenesis of ARDS caused by aspiration of hydrochloric acid, which could promote inflammation by directly activating pulmonary endothelial cells.

Optimization of reverse chemical ecology method: false positive binding of Aenasius bambawalei odorant binding protein 1 caused by uncertain binding mechanism.

Odorant binding proteins (OBPs) are considered as the core molecular targets in reverse chemical ecology, which is a convenient and efficient method by which to screen potential semiochemicals. Herein, we identified a classic OBP, AbamOBP1 from Aenasius bambawalei, which showed high mRNA expression in male antennae. Fluorescence competitive binding assay (FCBA) results demonstrated that AbamOBP1 has higher binding affinity with ligands at acid pH, suggesting the physiologically inconsistent binding affinity of this protein. Amongst the four compounds with the highest binding affinities at acid pH, 2, 4, 4-trimethyl-2-pentene and 1-octen-3-one were shown to have attractant activity for male adults, whereas (-)-limonene and an analogue of 1-octen-3-ol exhibited nonbehavioural activity. Further homology modelling and fluorescence quenching experiments demonstrated that the stoichiometry of the binding of this protein to these ligands was not 1: 1, suggesting that the results of FCBA were false. In contrast, the apparent association constants (Ka) of fluorescence quenching experiments seemed to be more reliable, because 2, 4, 4-trimethyl-2-pentene and 1-octen-3-one had observably higher Ka than (-)-limonene and 1-octen-3-ol at neutral pH. Based on the characteristics of different OBPs, various approaches should be applied to study their binding affinities with ligands, which could modify and complement the results of FCBA and contribute to the application of reverse chemical ecology.