OBJECTIVE To explore the mechanism of action of epigallocatechin-3-gallate(EGCG) in inhibiting laryngeal cancer cells. METHODS The expression of epidermal growth factor receptor(EGFR) in laryngeal cancer cell lines AMC-HN-8, TU686 and TU212 was detected by Western blotting, and the inhibitory effects of cetuximab and EGCG on three laryngeal cancer cells were detected by CCK-8 assay. A lentiviral vector containing EGFR promoter and Luc reporter gene was constructed to generate a TU686-EGFR-Luc cell line that could steadily express Luc activity. Luciferase assay was performed to evaluate the effect of EGCG on the transcription activity of EGFR promoter. Cell cycle and apoptosis of EGCG-treated laryngeal carcinoma cells were analyzed by flow cytometry, and changes of the levels of EGFR and downstream ERK1/2, cell cycle-associated proteins P53 and P27, apoptosis-associated proteins BCL2 and PART, and autophagy marker LC3A/B were further examined. RESULTS The laryngeal carcinoma cell lines were insensitive to cetuximab but could be effectively suppressed by EGCG. EGCG effectively inhibited the transcription activity of EGFR promoter. Treatment of TU686 cells at sub-IC50 dose EGCG resulted in significant cell cycle arrest at S phase with partial apoptosis. Significant inhibition of expression and activation of EGFR and downstream signaling pathway were observed. CONCLUSION EGCG can effectively downregulate EGFR and suppress laryngeal carcinoma cells, further investigation on in vivo effect and mechanisms are anticipated.

Objective:To comparatively study the similarities and differences between the clinical, pathological, and risk factors of advanced fibrosis in men and women with non-alcoholic fatty liver disease (NAFLD).Methods:267 patients with NAFLD diagnosed by liver biopsy were retrospectively included, and were divided into male and female groups. The difference of clinical and pathological indexes were compared between the two groups. The measurement data were in accordance with normal distribution. The comparison between the two groups was performed by independent sample t-test. The non-parametric test was used for non-normal distribution. The classification data were expressed as a percentage, and the chi-square test was used for comparison between groups. Logistic regression analysis was used to analyze the risk factors.Results:The age of onset of NAFLD was significantly lower in male than female patients ( P < 0.01). There was no statistically significant difference between the male and female groups in terms of body mass index and the prevalence of type 2 diabetes ( P > 0.05). Biochemical index: The levels of alanine aminotransferase, albumin, total bilirubin and uric acid were significantly higher in male than female patients ( P < 0.01). Liver pathology: The proportion of ballooning degeneration was significantly lower in male than female patients ( P < 0.01). There was not statistically significant difference between the two groups in the proportion of steatohepatitis score, non-alcoholic steatohepatitis (52.0% vs. 61.5%, P = 0.283) and advanced liver fibrosis (14.3% vs. 17.8%, P = 0.162). Thrombocytopenia was a common independent risk factor for advanced stage liver fibrosis ( OR = 0.984, 0.978~0.989, P < 0.01). Type 2 diabetes was only an independent risk factor for advanced stage liver fibrosis in men ( OR = 6.557, 1.667~25.782), P < 0.01). Elevated AST was only an independent risk factor for advanced stage liver fibrosis in women ( OR = 1.016, 1.003~1.028, P = 0.012). Conclusion:In NAFLD patients, there are some clinical and pathological differences between genders. Platelets are a common predictor of advanced liver fibrosis in men and women. Type 2 diabetes in men and elevated aspartate aminotransferase in women can be regarded as independent risk factors for advanced liver fibrosis.

Objective:To study the non-target metabolomics analysis and to analyze the metabolomic changesof cerebrospinal fluid (CSF) in patients with tuberculous meningitis.Methods:Case-control study. From July 2018 to July 2019, 20 cerebrospinal fluid specimens of diagnosed patients with tuberculous meningitis were collectedin the department of neurology from the first medical center of the PLA general hospital and the eighth medical center of the PLA general hospital and 20 CSF without tuberculous meningitis as the control. Among them, there were 12 males and 8 femalesin the tuberculous meningitis group, aged (37.9±16.1) years; there were 13 males and 7 femalesin the control group, aged (34.7±14.8) years. Using ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) technology with three different mode, namely reverse phase chromatography positive ion mode, reverse phase chromatography negative ion mode and hydrophilic chromatography positive ion mode,to detectthe metabolic fingerprints of patients′CSF and analyzed by SIMCA software for orthogonal partial least squares discriminant analysis (OPLS-DA). The variable importance projection value of OPLS-DA model (threshold value>1) plus the P value of t-test (P<0.05) was applied to find the differential metabolites in the cerebrospinal fluid of the two groups of patients.Results:Ten differential metabolites were found in CSF, including L-isoleucine, L-phenylalanine, L-kynurenine, L-methionine, L-tyrosine acid, dimethylglycine, L-alanine, L-threonine, L-histidine and L-lysine, and all of them were up-regulated in the tuberculous meningitis group.Conclusion:Changesof the amino acid metabolism found in the cerebrospinal fluid of tuberculous meningitis patients can provide basis for differential diagnosis and basic molecular research of tuberculous meningitis.

Objective Toinvestigatestatistically significant peptide peaks as biomarkersto diagnose osteoarticular tuberculosis, matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF MS) was applied to identify the characteristic fingerprint among the serum of patients with osteoarticular tuberculosis, rheumatoid arthritis and healthy adults.Methods Clinical Study. Serum samples of untreatedpatients with osteoarticular tuberculosis and rheumatoid arthritis were collected from August 2018 to December 2018, and serum samples of healthy adults from physical examination were collected as control. After analysis with MALDI-TOF MS, the serum peptide fingerprint datawas imported into software, and protein polypeptide peaks with obvious differences were screened to establish diagnostic models.Results Established the diagnostic model of osteoarticular tuberculosis and healthy adults with m/z 2943.9, 5929.6, 7615.4 and 9033.8 as differential protein polypeptides, the diagnostic model of osteoarticular tuberculosis and rheumatoid arthritis with m/z 4195.6, 5847.6, 5929.6 and 7748.6 as differential protein polypeptides. To these two models, the sensitivity were 95.00％ and 97.50％, respectively. The specificity were 85.71％ and 88.46％, respectively. The accuracy rates were 89.58％ and 92.39％, respectively. The AUC value of ROC curves were 0.8859 and 0.8709, respectively. Conclusions By mass spectrometry and software analysis, the serum protein polypeptides with statistical difference were found successfully. The related diagnostic modelsarealso established, which has certain reference value for auxiliary diagnosis of osteoarticular tuberculosis.

Objective To investigate the quality of life of army recruits in their basic military training period,and to analyze the influence of job burnout and work satisfaction on their quality of life.Methods A total of 625 recruits enrolled in 2014 of Xinjiang Army troop were chosen as subjects in this study by stratified cluster random sampling.Quality of life was assessed by Chinese version SF-36(Short-Form Health Survey Scale),job burnout was assessed by Chinese version Maslach Burnout inventory general survey (MBI-GS) and work satisfaction was assessed by Chinese version Minnesota Satisfaction Questionnaire (MSQ).Results The scores of PCS and MCS about 597 recruits were (85.93±12.62) and (81.10±14.12) respectively.According to demographic characteristics,the score of physiological function was lower than that of non-smoking group (t=2.009,P＜0.05),the score of role physical was lower than that of non-smoking group (t=2.617,P＜0.05),and the score of PCS was lower than that of non-smoking group (t=2.141,P＜0.05).As to except reported health transition,there were negative correlations between emotional exhaustion,depersonalization and other SF-36 scales (r=-0.344～-0.661,P＜0.01) respectively.There were negative correlations between personal accomplishment and vitality (r=-0.204,P＜0.05),role emotional (r=-0.239,P＜0.05),mental health (r=-0.289,P＜0.05) and MCS (r=-0.276,P＜0.05) respectively.Work satisfaction and quality of life was positively correlated (r=0.243～0.635,P＜0.01).As to the independent variable,regression analysis showed that emotional exhaustion (β=-4.732,P＜0.01),Minnesota external satisfaction (β=0.783,P＜0.01) influenced PCS,the difference was statistically significant.Emotional exhaustion(β=-6.534,P＜0.01),pre-enlistment place of residence (β=-5.319,P＜ 0.05),Minnesota external satisfaction (β=0.813,P＜0.01)influenced MCS,the difference was statistically significant.Conclusion Job burnout and work satisfaction influence the quality of life about the army recruits,the more job burnout signify the lower quality of life,and its influence on mental health is more significant.The higher work satisfaction signify the higher quality of life.

Objective: To investigate the fatigue status of military personnel stationed in plateau and high cold region, and to analyze the mediator effect of trait coping style on job stress and fatigue. Methods: In October 2010, with the method of cluster random sampling survey, 531 military personnel stationed in plateau and high cold region were chosen as subject. The fatigue status were evaluated by the Chinese version multidimensional fatigue inventory (MFI-20) , job stress were evaluated by the Job Stress Survey (JSS) , and trait coping style were evaluated by the Trait Coping Style Questionnaire (TCSQ) . Results: According to the information of different population characteristics, mean rank of physical fatigue about the urban (town) group were higher than that of rural group (Z=-2.200, P<0.05) ; mean rank of reduced motivation about the urban (town) group were higher than that of rural group (Z=-2.781, P<0.05) ; mean rank of general fatigue scores about the urban (town) group were higher than that of rural group (Z=-3.026, P<0.05) ; mean rank of physical fatigue about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-4.045, P<0.05) ; mean rank of reduced motivation about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-2.182, P<0.05) ; mean rank of mental fatigue about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-2.879, P<0.05) ; mean rank of general fatigue scores about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-3.647, P<0.05) ; mean rank of reduced motivation were significant statistical difference among the military officers, sergeancy and soldier group (F=18.965, P<0.05) ; mean rank of general fatigue scores were significant statistical difference among the military officers, sergeancy and soldier group (F=14.711, P<0.05) . The score of negative coping style were positively correlated with the score of physical fatigue (r_s=0.129) , reduced activity (r_s=0.123) , reduced motivation (r_s=0.149) and general fatigue (r_s=0.174) respectively, the score of organizational support lack strength were positively correlated with the score of physical fatigue (r_s=0.090) , reduced activity (r_s=0.098) , reduced motivation (r_s=0.099) and general fatigue (r_s=0.130) respectively. The mediator effect of negative coping style on the job stress and fatigue was 0.013 (P<0.01) . Conclusion The fatigue statuses of the urban (town) group and the up or equal 20-years old age group are poor, and the negative coping style plays mediator effect on the job stress and fatigue.

We discussed the function of four pairs of genes in the toxin-antitoxin system of Mycobacterium tuberculosis,providing theoretical foundation and scientific basis for studying the transmission mechanism of Mycobacterium tuberculosis.Four pairs of genes which belong to VapBC family,including four VapC genes (Rv1720c,Rv2103c,Rv2494,Rv3408) and four VapB genes (Rv1721c,Rv2104c,Rv2493,Rv3407) were chosen.We constructed a serial of arabinose-induced hybrid plasmid system in Escherichia coli and a serial of acetamide-induced hybrid plasmid system in Mycobacterium smegmatis respectively,in order to observe the potential inhibition effect of VapC and the release inhibition of homologous VapB.Results showed that only one toxin gene(Rv2103c) showed the function of bacteriostasis in both E.coli and M.smegmatis and the homologous antitoxin gene(Rv2104c) could release the inhibition of growth.We built the inducible systems of VapBC family in both E.coli and M.smegmatis respectively and found only a pair of toxin and antitoxin genes(Rv2103c,Rv2104c) had the function of inhibition and release for the growth of bacteria.And two pairs of toxin genes(Rv1720c,Rv2494) did not have the function of inhibition for the growth of both E.coli and M.smegmatis.Whereas,another toxin gene VapC47(Rv3408) also did not have the bacteriostastic activity,only this result was not consistent with the existing literature.We speculated that the reason for this kind of difference may be the different inducible systems we used.Cause the other three results were consistent with all existing literature and the doubtful result also appeared in other reports,so our protocol could be confirmed as reliable,and we would use it to build inducible systems and make further functional identification of certain toxin and antitoxin genes that we are interested in.

Abstract Gram-negative ( G-) bacteria, such as denitrifying bacteria and anaerobic ammonia oxidation bacteria, are highly social organisms capable of sophisticated cooperative behavior mediated via quorum sensing. As signal molecules of the chemical communication, N-acyl-homoserine lactones ( AHLs ) can mediate the quorum sensing of the functional microbial population and regulate the population density. To understand the growth of functional microbial population and the mechanism for biological nitrogen removal in upflow microaerobic sludge reactors ( UMSRs ) treating organic wastewater with low ratio of chemical oxygen demand to total nitrogen, a method was established to simultaneously detect AHLs in the microaerobic processes. Water-sludge mixtures sampled from the UMSRs were pretreated in sequence by liquid-liquid extraction using ethyl acetate, rotary evaporation, constant volume with methanol, separation by C18 column. Gradient elution was carried out using 5 mmol/L ammonium acetate ( containing 0 . 1% formic acid ) and methanol as mobile phases. On the base of multiple reaction monitoring analysis, a triple quadrupole mass spectrometer with an electrospray ionization was introduced to detect the target compounds. Nine kinds of AHLs were used to evaluate the established method and the results showed that the detection limits were 0 . 01-0 . 5 μg/L and all of the AHLs presented excellent linearity with the concentration ranging from 0 . 5 to 100 μg/L. The recovery and relative standard deviation ranged from 62. 5% to 118. 1% and 2. 9% to 12. 1%, respectively. The analysis could be finished within 6. 5 min. The rapid, accurate and precise method for detecting AHLs provided a new insight into the growth and metabolic activity of functional microbial population in the activated sludge processes to understand the mechanism of biological nitrogen removal, suggesting a good application in regulation and operation of wastewater biological treatment processes.

Objective To explore the effects of job content on psychological stress in young recruits.Methods In October 2014,625 young recruits enrolled in one troop of Xinjiang Military Command in 2014 were chosen as subjects by multi-stage stratified random sampling.The Chinese version of the job content questionnaire (JCQ) and the psychological stress self evaluation test (PEST) were used to investigate the subjects.The subjects were divided into two groups with scores higher and lower than the mean score of three subscales (job requirement,degree of autonomy,and social support) of JCQ to explore the effects of job content on psychological stress in young recruits.The correlation of psychological stress with three subscales of job content was evaluated using the Pearson's correlation analysis.Binary logistic regression analysis was used to analyze the influencing factors for psychological stress.Results The PEST score of young recruits was 49.98± 9.98.Forty-five (7.68％) out of them had scores of ≥70 points and were diagnosed with high levels of psychological stress.When the subjects were grouped based on socio-demographic characteristics,a high level of psychological stress was significantly more frequent in subjects less than 20 years of age than in those not less than 20 years of age,in smoking subjects than in non-smoking subjects,and in urban residents than in rural residents (10.42％ vs 5.03％,P＜0.05;10.14％ vs 5.33％,P＜0.05;10.85％ vs 5.88％,P ＜0.05).There were no significant differences in educational level or identity before recruitment between subjects with different levels of psychological stress (P＞0.05).In various job content domains that had impacts on psychological stress,subjects with a low score of social support had significantly higher PEST scores than those with a high score of social support (50.96±10.35 vs 48.49±9.22,P＜0.01).The PEST score was negatively correlated with the degree of autonomy and social support(r=-0.103,P＜0.05;r=-0.239,P＜0.05).The results of logistic regression analysis showed that job requirement and social support were influencing factors for psychological stress (OR =0.718,95％CI=0.718 (0.607～0.851),P＜0.05;OR=1.185,95％CI=1.185 (1.087～1.292),P＜0.05).Conclusion Psychological stress of young recruits in army is not optimistic.Psychological stress is weakly negatively correlated with social support.The low scores on job requirement subscale and social support subscale may be potential protective factor and risk factor for psychological stress,respectively.

Objective To explore the effects of job content on psychological stress in young recruits.Methods In October 2014,625 young recruits enrolled in one troop of Xinjiang Military Command in 2014 were chosen as subjects by multi-stage stratified random sampling.The Chinese version of the job content questionnaire (JCQ) and the psychological stress self evaluation test (PEST) were used to investigate the subjects.The subjects were divided into two groups with scores higher and lower than the mean score of three subscales (job requirement,degree of autonomy,and social support) of JCQ to explore the effects of job content on psychological stress in young recruits.The correlation of psychological stress with three subscales of job content was evaluated using the Pearson's correlation analysis.Binary logistic regression analysis was used to analyze the influencing factors for psychological stress.Results The PEST score of young recruits was 49.98± 9.98.Forty-five (7.68％) out of them had scores of ≥70 points and were diagnosed with high levels of psychological stress.When the subjects were grouped based on socio-demographic characteristics,a high level of psychological stress was significantly more frequent in subjects less than 20 years of age than in those not less than 20 years of age,in smoking subjects than in non-smoking subjects,and in urban residents than in rural residents (10.42％ vs 5.03％,P＜0.05;10.14％ vs 5.33％,P＜0.05;10.85％ vs 5.88％,P ＜0.05).There were no significant differences in educational level or identity before recruitment between subjects with different levels of psychological stress (P＞0.05).In various job content domains that had impacts on psychological stress,subjects with a low score of social support had significantly higher PEST scores than those with a high score of social support (50.96±10.35 vs 48.49±9.22,P＜0.01).The PEST score was negatively correlated with the degree of autonomy and social support(r=-0.103,P＜0.05;r=-0.239,P＜0.05).The results of logistic regression analysis showed that job requirement and social support were influencing factors for psychological stress (OR =0.718,95％CI=0.718 (0.607～0.851),P＜0.05;OR=1.185,95％CI=1.185 (1.087～1.292),P＜0.05).Conclusion Psychological stress of young recruits in army is not optimistic.Psychological stress is weakly negatively correlated with social support.The low scores on job requirement subscale and social support subscale may be potential protective factor and risk factor for psychological stress,respectively.