AIM:To investigate the effects of cigarette smoke exposure during adolescence on the inflammato-ry response mediated by microglia in the prefrontal cortex of mice,and its impact on learning and memory functions in adulthood.METHODS:72 two-week-old healthy male Kunming mice,with each weighing(11.0±1.5)g,were random-ly divided into control and cigarette exposure groups(n=36 per group).The mice in the cigarette exposure group were pas-sively exposed to 6 cigarettes daily for 10 weeks.At three time points of 4-week-old(infancy),8-week-old(adoles-cence),and 12-week-old(adulthood),six mice were selected from each group to have their neurobehavioral and patholog-ical changes examined.In particular,the step-down test,three-chamber social interaction test,and novel object recogni-tion test were used to detect changes in learning and memory abilities and cognitive behavior.Immunofluorescence testing was performed to detect the morphology,number of synapses,and expression of inflammatory factor apoptosis-associated speck-like protein containing a CARD(ASC)around the microglial cells in the prefrontal cortex of mice in each group.Western blot was performed to assess the expression levels of synaptophysin(SYP)and postsynaptic density protein-95(PSD-95)in the cerebral cortex of mice in each group.Enzyme-linked immunosorbent assay(ELISA)was performed to detect the protein expression of tumor necrosis factor-α(TNF-α),interleukin 1β(IL-1β),and IL-6 in the prefrontal cor-tex.RESULTS:(1)In the step-down test,the latency of mice at 8 and 12 weeks of age was significantly shortened,and the number of errors was significantly increased in the cigarette exposure group compared with the age-matched control group(P＜0.01).In addition,the social recognition time and exploration time for novel objects were prolonged(P＜0.05).(2)Immunofluorescence assays revealed that exposure to cigarette smoke in mice,at both 8 and 12 weeks of age,resulted in a reduction of SYP-positive puncta within the prefrontal cortex.Concurrently,there was an observed increase in the number of Iba1-positive microglia,which exhibited an activated phenotype,as well as an elevation in ASC-positive puncta in proximity to the microglia.Western blot further revealed reduced expression of synaptophysin protein SYP and PSD-95 in the cerebral cortex of the mice at 8 and 12 weeks of age in the cigarette exposure group(P＜0.05).(3)ELISA showed increased levels of inflammatory factors TNF-α,IL-1β,and IL-6 in the prefrontal cortex of the mice at 8 and 12 weeks of age in the cigarette exposure group(P＜0.05).CONCLUSION:Exposure to cigarette smoke during adoles-cence in mice may result in the enhanced secretion of inflammatory factors through the activation of microglia in the pre-frontal cortex.This activation can alter microglial function and induce synaptic damage,consequently impairing learning,memory,and cognitive abilities in adulthood.

AIM:To investigate the effects of cigarette smoke exposure during adolescence on the inflammato-ry response mediated by microglia in the prefrontal cortex of mice,and its impact on learning and memory functions in adulthood.METHODS:72 two-week-old healthy male Kunming mice,with each weighing(11.0±1.5)g,were random-ly divided into control and cigarette exposure groups(n=36 per group).The mice in the cigarette exposure group were pas-sively exposed to 6 cigarettes daily for 10 weeks.At three time points of 4-week-old(infancy),8-week-old(adoles-cence),and 12-week-old(adulthood),six mice were selected from each group to have their neurobehavioral and patholog-ical changes examined.In particular,the step-down test,three-chamber social interaction test,and novel object recogni-tion test were used to detect changes in learning and memory abilities and cognitive behavior.Immunofluorescence testing was performed to detect the morphology,number of synapses,and expression of inflammatory factor apoptosis-associated speck-like protein containing a CARD(ASC)around the microglial cells in the prefrontal cortex of mice in each group.Western blot was performed to assess the expression levels of synaptophysin(SYP)and postsynaptic density protein-95(PSD-95)in the cerebral cortex of mice in each group.Enzyme-linked immunosorbent assay(ELISA)was performed to detect the protein expression of tumor necrosis factor-α(TNF-α),interleukin 1β(IL-1β),and IL-6 in the prefrontal cor-tex.RESULTS:(1)In the step-down test,the latency of mice at 8 and 12 weeks of age was significantly shortened,and the number of errors was significantly increased in the cigarette exposure group compared with the age-matched control group(P＜0.01).In addition,the social recognition time and exploration time for novel objects were prolonged(P＜0.05).(2)Immunofluorescence assays revealed that exposure to cigarette smoke in mice,at both 8 and 12 weeks of age,resulted in a reduction of SYP-positive puncta within the prefrontal cortex.Concurrently,there was an observed increase in the number of Iba1-positive microglia,which exhibited an activated phenotype,as well as an elevation in ASC-positive puncta in proximity to the microglia.Western blot further revealed reduced expression of synaptophysin protein SYP and PSD-95 in the cerebral cortex of the mice at 8 and 12 weeks of age in the cigarette exposure group(P＜0.05).(3)ELISA showed increased levels of inflammatory factors TNF-α,IL-1β,and IL-6 in the prefrontal cortex of the mice at 8 and 12 weeks of age in the cigarette exposure group(P＜0.05).CONCLUSION:Exposure to cigarette smoke during adoles-cence in mice may result in the enhanced secretion of inflammatory factors through the activation of microglia in the pre-frontal cortex.This activation can alter microglial function and induce synaptic damage,consequently impairing learning,memory,and cognitive abilities in adulthood.

Objective:To investigate the clinicopathological features of basal cell layer type high-grade squamous dysplasia of the esophagus.Methods:Fifty-two cases of basal cell layer type high-grade squamous dysplasia of the esophagus were collected at PLA Joint Logistics Support Force 989 Hospital (34 cases) and Beijing Chaoyang Hospital (18 cases) from 2009 to 2019. The clinical, histological and immunohistochemical features were characterized. Related literature was also reviewed.Results:The median age of the 52 patients was 64 years (range 43-72 years). There were 35 men and 17 women, with a male to female ratio of 2.1∶1.0. There were 8 cases in the upper esophagus, 41 in the middle esophagus and 3 in the lower esophagus. According to the Paris Classification, 24 cases were 0-Ⅱb and 28 cases were 0-Ⅱc. Endoscopic examination showed that the color of the lesions was red and the edge was irregular. The narrow band imaging showed that the lesions were brown, and the microvascular abnormalities on the mucosal surface were observed with high magnification. Iodine staining of the lesions showed no or light staining and irregular border. Histologically, the basal layer of squamous epithelium was hypercellular, with large and hyperchromatic nuclei, and disordered cell arrangement. A high proportion of the cases showed a down-growth pattern and associated invasive squamous cell carcinoma. The immunohistochemical staining of 37 cases showed that the mutation rate of p53 was 48.6% (18/37), the median of Ki-67 labeling index was 60% (range 20%-90%), the median of Ki-67 labeling index of the basal tumor cells was 26/HPF (range 5-70/HPF), and the rate of abnormal Ki-67 distribution pattern was 37(100.0%). According to the initial pathological diagnosis, there were 8 cases of low-grade intraepithelial neoplasia, 2 cases of atypical epithelial cells and 42 cases of high-grade intraepithelial neoplasia.Conclusions:The basal cell layer type high-grade squamous dysplasia of the esophagus has a unique morphology. The dysplasia is mainly limited to the lower half part of the squamous epithelium. With marked cytological atypia and prominent invasiveness pattern, it is likely to develop into invasive squamous cell carcinoma at an early stage of the disease. The rate of pathologic misdiagnosis (such as low-grade lesion) is high. The p53 mutation and Ki-67 abnormal distribution pattern are helpful features for confirming the diagnosis of such high-grade dysplasia.

Objective To investigate the expressions of Th17 lymphocytes and interleukin-17 (IL-17) in peripheral blood of patients with non-small cell lung cancer (NSCLC) and its clinical significance. Methods Sixty patients with primary and untreated NSCLC were enrolled and designed as experimental group, at the same time, 60 healthy volunteers were collected as control group. Flow cytometry (FCM) was used to detect the level of Th17 lymphocytes. Enzyme linked immunosorbent assay (ELISA) was used for detecting the level of IL-17. The relationship between the expression levels of Th17 and IL-17 in peripheral blood and clinicopathological features was compared between the two groups. Results The peripheral blood levels of Th17 lymphocytes and IL-17 in the experimental group [(1.7±1.2) ％, (8.3±2.5) pg/ml] were higher than those in the control group [(0.9 ±0.6) ％, (5.4 ±1.2) pg/ml] (P< 0.05). The peripheral blood expression of Th17 lymphocytes and IL-17 in patients with smoking history [(1.8±1.2) ％, (8.8±3.7) pg/ml] were higher than those in patients without smoking history [(1.6±1.2)％, (8.0±2.2) pg/ml], and the peripheral blood expression of Th17 lymphocytes and IL-17 were higher in patients with squamous-cell carcinoma [(1.8 ±1.2) ％, (9.4 ±4.7) pg/ml] than those in patients with adenocarcinoma [(1.6±1.1) ％, (7.3±3.9) pg/ml], furthermore, they were also higher in patients with stage Ⅲ-Ⅳ than those in patients with stage Ⅰ-Ⅱ (P < 0.05). Conclusion Th17 lymphocytes and IL-17 play certain roles in the occurrence and progression of NSCLC.

Objective To explore the value of dual energy CT imaging in the diagnosis of gouty tophus deposition.Methods Sixty-five cases of patients with gout treated in this hospital between December 2012 to December 2015 were selected.Another 65 cases of patients(rheumatoid arthritis,degenerative osteoarthritis and unexplained leg pain) were selected as control group.The re sults of dual energy CT imaging between two group were compared.Three experienced clinicians who didn't understand the imaging results were selected.They had a medical history and physical examination in patients with gout.The differences between clinical evaluation and imaging results were compared.Results Dual energy CT imaging showed that there were no tophi deposition in con trol group;the topi deposition in wrist joint,knee joint and ankle joint were 6.06％oo (33/34),100.00％ (22/22) and 96.49％ (55/57) in gout group.Dual energy CT imaging detected a total of 545 parts of tophus deposition.It was 4.36 times of the result of clinical evaluation(125 parts).Conclusion Dual energy CT imaging shows the advantages of simple noninvasive,high accuracy and low rate of misdiagnosis in diagnosing gout nodules.

Objective To clone and express Mycobacterium tuberculosis fusion antigen ESAT-6 and CFP-10 ( EC) and to evaluate the biological activity of the fusion antigen EC in inducing specific cytokines secretion from THP -1 cells. Methods The fusion antigen EC gene was cloned into pET-30 a prokaryotic expression vector and expressed highly in E.coli BL21.Then, the THP-1 cells were stimulated with purified fusion antigen EC of different concentrations (10 and 20 μg/ml).Culture supernatants were collected after 12 h and 24 h, respectively.The secretion levels of IL-2, IL-4, IL-6, IL-8, IL-10, GM-CSF, TNF-αand IFN-γin THP-1 cell culture supernatants were detected using Bio-PlexProTM Assays kit.Results The M.tuberculosis fusion antigen EC was cloned and expressed successfully .The secretion levels of IL-6, IL-8 and TNF-αin EC infected THP-1 cells were significantly higher than those in THP-1 cells (P<0.05).The secretion levels of other cytokines did not change significantly .Conclusion The obtained M.tuberculosis fusion antigen EC has biological activity in inducing the THP-1 cells to secrete a higher level of IL-6,IL-8 and TNF-α.

Objective To investigate the correlation between urinary 8-iso-prostaglandin F2α (8-iso-PGF2α) level and carotid atherosclerotic plaque stability in patients with acute ischemic stroke.Methods The patients with acute ischemic stroke were enrolled consecutively.According to the findings of carotid artery ultrasound,they were divided into either a non-plaque group or a plaque group,and then the plaque group was redivided into a stable plaque subgroup and an unstable plaque subgroup.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of urinary 8-iso-PGF2α.Multivariate logistic regression analysis and Spearman correlation analysis were used to investigate the relationship between 8-iso-PGF2α and carotid artery plaques.Results A total of 150 patients with acute ischemic stroke were enrolled,including 104 had carotid artery plaque (30 were stable plaques and 74 were unstable plaques) and 46 had no carotid artery plaque.The urinary 8-iso-PGF2α level (86.45 ± 6.20 ng/mmol creatinine vs.45.70 ±6.19 ng/mmol creatinine; t =37.136,P=0.001) and intima-media thickness (IMT) (2.89 ± 1.03 mm vs.0.86 ±0.53 mm; t =3.518,P =0.002) in the plaque group were significantly higher than those in the nonplaque group.Multivariate logistic regression analysis showed that urinary 8-iso-PGF2α level (odds ratio [OR] 1.183,95％ confidence interval [CI] 1.087-1.276; P=0.001) and IMT (OR 28.642,95％ CI 8.276-137.231; P =0.001) were the independent risk factors for carotid artery plaque.Pearson correlation analysis showed that there was no significant correlation between urinary 8-iso-PGF2α level and carotid artery IMT (r =0.075,P =0.264).The urinary 8-iso-PGF2α level of the unstable plaque subgroup was significantly higher than that of the stable plaque subgroup (97.30 ± 7.20 ng/mmol creatinine vs.69.17 ±9.25 ng/mmol creatinine; t =16.506,P =0.001).Multivariate logistic regression analysis showed that urinary 8-iso-PGF2α level (OR 4.652,95％ CI 1.732-12.643; P =0.001) was an independent risk factor for unstable plaque.Conelusions The urinary 8-iso-PGF2α level is associated with the existence of carotid atherosclerotie plaque and instability,its increased level is an independent risk factor for carotid plaque instability in patients with ischemic stroke.

Objective To investigate the application of strain rate imaging in quantitative assessment of left ventricular regional diastolic function after intracoronary stent implantation.Methods Fifty-six healthy person and 60 patients with coronary artery disease were performed quantitative assessment of left ventricular regional diastolic function by apical four chamber view,apical two chamber view and left ventricular long axis view before and after intracoronary stent implantation using strain rate imaging.Results Compared with those of normal myocardium,the peaks of strain rate curve at diastole of ischemic myocardium were reduced,and were increased after operation (P ＜ 0.05).Conclusions Intracoronary stent implantation can significantly improve the blood supply to the ischemia myocardium.Strain rate imaging can quantitatively analyze the changes of left ventricular regional diastolic function.

Objective To explore the feasibility of inherent depressive Fawn-Hooded (FH/Wjd)rats as a comorbidity model of mental disorder and irritable bowel syndrome (IBS) by comparing visceral sensitivity of FH/Wjd rats and Sprague-Dawley (SD) rats. Methods Depression trait of FH/Wjd rats was validated through forced swimming test (FST) and sucrose preference test (SPT).Visceral sensitivity to colorectal distention (CRD) under various pressures was assessed by abdominal withdrawal reflex (AWR). The expression of 5-HT in rats' colon,c-fos expression in colon and brain areas of infralimbic cortex (IL),prelimbic cortex (PrL) and rostral anterior cingulated cortex (rACC)was tested with immunohistochemistry. Results FST indicated that the immobility time of FH/Wjd rats was significantly longer than that in SD rats (t=-8. 931,P＜0. 01). SPT showed that the ratio of sucrose water in total liquid consumed was significantly lower in FH/Wjd rats than that of SD rats(t=4. 155,P=0. 01). At each CRD pressure gradient (20,40,60 and 80 mm Hg,1 mm Hg=0. 133kPa),AWR score was all significantly higher in FH/Wjd rats than that of SD rats (t=-2. 697,-3.464,-6.822 and -3. 976,P=0. 022,0.006,0.000 and 0.003). The expression level of 5-HT in FH/Wjd rats' control group was significantly higher than that of SD rats' control group (t=-11.371,P=0. 000). Compared with their own control group,5-HT expression increased both in FH/Wjd rats dilated groups and SD rats dilated groups,and the expression level in FH/Wjd rats was higher than that of SD rats (t= -3. 364,P= 0. 007). There was no significant difference of c-fos expression in colon and prefrontal brain areas between FH/Wjd rats' control group and SD rats'control group (colon:t=-0. 129,P= 0. 900;IL:t=-1. 316,P= 0. 218;PrL:t=1. 241,P = 0. 243;rACC:t=2. 151,P = 0. 057). Compared with their own control group,the expression of c-fos in colon and prefrontal brain areas significantly increased both in FH/Wjd rats dilated groups and SD rats dilated groups,and the expression level in FH/Wjd rats was higher than that of SD rats (colon: t=- 5.864,P= 0.000;IL:t=-2. 530,P = 0. 030;PrL:t=-7. 039,P= 0. 000;rACC:t=-6. 489,P=0. 000). Conclusions Inherent depressive FH/Wjd rats present visceral hypersensitivity. Meanwhile,the expression of 5-HT in colon is high,and with hyperreactivity to visceral noxious stimuli in colon and brain IL,PrL,and rACC areas. FH/Wjd rats might be a comorbidity animal model of mental disorder and IBS.

Objective To apply fluo-rescencc-activated cell sorting(FACS) in sorting T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+), which establish the separating method of series of cells from human peripheral blood, so that the scientific research and clinical research could be carried out were specifically. Methods 10 cases of normal peripheral blood were collected and T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) were stained with florescence conjugated antibodies. The positive cells were sorted by FACS. Results Before sorting the peripheral blood, the proportion of the T lymphocyte(CD_3~+) and granulocyte (CD_(15)~+) in the leukocyte is 48.8 % and 30.8 %; after sorting by FACS, the purity of T lymphocytes (CD_3~+) is up to 98 % and the recovery is about 95 %; the purity of granulocyte (CD_(15)~+) is up to 97 % and the recovery is about 96 %. Conclusion The FACS could Mlow us to quickly sort T lymphocyte (CD_3~+) and granulocyte (CD_(15)~+) with higher recovery and higher purity from the peripheral blood.