Objective To investigate the effects of Forkhead box protein O3a(FoxO3a)on M2 polarization and ferroptosis of macrophages in allergic rhinitis(AR).Methods Nasal lavage fluids were collected from 30 patients with AR and 30 healthy volunteers,while nasal mucosa samples were obtained from 3 AR patients and 3 healthy volunteers.The levels of FoxO3a in nasal lavage fluids were detected by ELISA.M0 macrophages were transfected with siRNA negative control plasmid(si-Con),siRNA FoxO3a plasmid(si-FoxO3a),pcDNA3.1-vector,or pcDNA3.1-FoxO3a plasmid,and then induced to polarize towards M2 type by interleukin-4(IL-4).M0 macrophages were set as the control group,IL-4 model group,IL-4+si-Con group,IL-4+si-FoxO3a group,IL-4+si-FoxO3a+nuclear factor erythroid 2-related factor 2(Nrf2)inhibitor ML385 group,IL-4+pcDNA3.1-vector group,and IL-4+pcDNA3.1-FoxO3a group.A co-culture system of M0 macrophages and human nasal epithelial cells(NECs)was established,and then divided into the NECs+M0 macrophages(control group),NECs+M2 macrophages(IL-4 model group),NECs+si-FoxO3a-transfected macrophages(IL-4+si-FoxO3a group)and NECs+si-FoxO3a-transfected macrophages+ML385(IL-4+si-FoxO3a+ML385 group).Apoptosis of NECs was detected by flow cytometry.C57BL/6 mice were divided into normal group,AR model group and shRNA-FoxO3a adeno-associated virus(AAV-sh-FoxO3a)group.HE staining was used to observe histopathological changes in nasal mucosa tissue,and flow cytometry was used to detect the proportion of M2 macrophages in mouse nasal lavage fluids.The levels of IL-5,IL-13 and NECs mucin 5AC(MUC5AC)in mouse nasal lavage fluids were detected by ELISA;Western blot was used to detect the levels of FoxO3a,IL-10,arginase 1(Arg-1),Nrf2,heme oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4).Results Compared with the normal,the expression of FoxO3a was increased in nasal lavage fluids and nasal mucosa tissues of AR patients.Compared with the control group,the proportion of M2 macrophages and the levels of FoxO3a were increased in the IL-4 model group,while the expression levels of Nrf-2,HO-1 and GPX4 were decreased,and the levels of Fe2+and malondialdehyde(MDA)were increased(P<0.05).Compared with the IL-4 model group,these indicators were reversed in the IL-4+si-FoxO3a group.Compared with the IL-4 model group,the proportion of M2 macrophages was increased in the IL-4+pcDNA3.1-FoxO3a group.In the co-culture system,compared with the IL-4 model group,the apoptosis of NECs and MUC5AC secretion were decreased in the IL-4+si-FoxO3a group(P<0.05).ML385 reversed the protective effect of si-FoxO3a on NECs.Mice in the AAV-sh-FoxO3a group showed reduced nasal mucosa tissue damage,decreased levels of IL-5 and IL-13 in nasal lavage fluids,and a lower number of M2 macrophages,decreased level of FoxO3a in M2 macrophages,while increased expression of GPX4,Nrf2 and HO-1.Conclusion Interference with FoxO3a protein expression can inhibit macrophage ferroptosis,suppress M2 polarization,and reduce NECs damage,which is related to the activation of the Nrf-2/HO-1 signaling pathway in macrophages.

Objective To investigate the effects of Forkhead box protein O3a(FoxO3a)on M2 polarization and ferroptosis of macrophages in allergic rhinitis(AR).Methods Nasal lavage fluids were collected from 30 patients with AR and 30 healthy volunteers,while nasal mucosa samples were obtained from 3 AR patients and 3 healthy volunteers.The levels of FoxO3a in nasal lavage fluids were detected by ELISA.M0 macrophages were transfected with siRNA negative control plasmid(si-Con),siRNA FoxO3a plasmid(si-FoxO3a),pcDNA3.1-vector,or pcDNA3.1-FoxO3a plasmid,and then induced to polarize towards M2 type by interleukin-4(IL-4).M0 macrophages were set as the control group,IL-4 model group,IL-4+si-Con group,IL-4+si-FoxO3a group,IL-4+si-FoxO3a+nuclear factor erythroid 2-related factor 2(Nrf2)inhibitor ML385 group,IL-4+pcDNA3.1-vector group,and IL-4+pcDNA3.1-FoxO3a group.A co-culture system of M0 macrophages and human nasal epithelial cells(NECs)was established,and then divided into the NECs+M0 macrophages(control group),NECs+M2 macrophages(IL-4 model group),NECs+si-FoxO3a-transfected macrophages(IL-4+si-FoxO3a group)and NECs+si-FoxO3a-transfected macrophages+ML385(IL-4+si-FoxO3a+ML385 group).Apoptosis of NECs was detected by flow cytometry.C57BL/6 mice were divided into normal group,AR model group and shRNA-FoxO3a adeno-associated virus(AAV-sh-FoxO3a)group.HE staining was used to observe histopathological changes in nasal mucosa tissue,and flow cytometry was used to detect the proportion of M2 macrophages in mouse nasal lavage fluids.The levels of IL-5,IL-13 and NECs mucin 5AC(MUC5AC)in mouse nasal lavage fluids were detected by ELISA;Western blot was used to detect the levels of FoxO3a,IL-10,arginase 1(Arg-1),Nrf2,heme oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4).Results Compared with the normal,the expression of FoxO3a was increased in nasal lavage fluids and nasal mucosa tissues of AR patients.Compared with the control group,the proportion of M2 macrophages and the levels of FoxO3a were increased in the IL-4 model group,while the expression levels of Nrf-2,HO-1 and GPX4 were decreased,and the levels of Fe2+and malondialdehyde(MDA)were increased(P<0.05).Compared with the IL-4 model group,these indicators were reversed in the IL-4+si-FoxO3a group.Compared with the IL-4 model group,the proportion of M2 macrophages was increased in the IL-4+pcDNA3.1-FoxO3a group.In the co-culture system,compared with the IL-4 model group,the apoptosis of NECs and MUC5AC secretion were decreased in the IL-4+si-FoxO3a group(P<0.05).ML385 reversed the protective effect of si-FoxO3a on NECs.Mice in the AAV-sh-FoxO3a group showed reduced nasal mucosa tissue damage,decreased levels of IL-5 and IL-13 in nasal lavage fluids,and a lower number of M2 macrophages,decreased level of FoxO3a in M2 macrophages,while increased expression of GPX4,Nrf2 and HO-1.Conclusion Interference with FoxO3a protein expression can inhibit macrophage ferroptosis,suppress M2 polarization,and reduce NECs damage,which is related to the activation of the Nrf-2/HO-1 signaling pathway in macrophages.

Objective:To explore the method of establishing a modified demyelination and myelination regeneration model induced by dicyclohexanone oxalyl dihydrazone (CPZ) in mice with multiple sclerosis (MS), and to analyze the image markers of demyelination and myelination regeneration in mouse MS model.Methods:After the intragastrically administered with sodium carboxymethyl cellulose (CMCNa) for one week, a total of 30 C57BL/6 male mice were randomly divided into the control group ( n=10), the demyelination group ( n=10), and the remyelination group ( n=10). The mice of the control group were immediately performed MR scanning and pathological specimen obtaining; the mice in the demyelination group were administered with intragastrical CPZ-CMCNa once a day for 6 weeks for inducing demyelination, then received MR scanning and specimen obtaining with the same protocols used in control group; the mice in the remyelination group were administered with intragastrical CPZ-CMCNa once a day for six weeks for demyelination, then CPZ was withdrawn and normal diet was given for another four weeks. Then MR scanning and specimen obtaining were performed with the same protocols used in the other two groups. Regions of interest (ROIs) were set at the rostrum of corpus callosum (rCC), the bilateral normal appearing white matters (NAWM) of the rostrum of corpus callosum, and the bilateral cerebral cortex (Cx). The normalized T 2WI (T 2-normalized), fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) values were compared among the three groups by one-way ANOVA. Results:The demyelination and remyelination mice model of MS were successfully established. The T 2-normalized values of rCC in control group, demyelination group and remyelination group were 0.47±0.03, 0.72±0.04, 0.54±0.04, respectively, with statistically significant difference found ( F=90.511, P<0.05). Post-hoc multiple comparisons showed significant differences among those groups ( P<0.05). There was no significant difference of T 2-normalized value in NAWM and Cx among the three groups ( P>0.05). Moreover, there were significant differences in the FA values (0.36±0.04, 0.29±0.03, and 0.32±0.05), the MD values [(0.572±0.015), (0.598±0.034), and (0.626±0.043)×10 -3 mm 2/s], the AD values [(0.79±0.04), (0.77±0.06), and (0.83±0.04)×10 -3 mm 2/s], and the RD values [(0.46±0.02), (0.51±0.03), and (0.53±0.05)×10 -3 mm 2/s] of rCC of the control group, the demyelination group, and the remyelination group (all P<0.05). Significant difference was found in FA values between the demyelination group and the control group ( P<0.05), and in MD values between the remyelination group and the control group ( P<0.05), as well as in AD values between the remyelination group and the demyelination group ( P<0.05). There were also significant differences in RD values between the remyelination group and the control group, and the demyelination group and the control group (all P<0.05). However, no significant difference was found in all diffusion tensor imaging (DTI) metrics of NAWM and Cx among the three groups (all P>0.05). The LFB-eosin staining showed that the myelin sheath of rCC was lost in the demyelination group, and the rCC was partially regenerated and repaired in the remyelination group. Conclusion:The modified CPZ-CMCNa model can selectively induce demyelination and remyelination of rCC, and the changes of demyelination and remyelination of rCC in the modified CPZ-CMCNa model can be quantitatively detected by T 2WI combined with DTI, which might provide related theoretical basis for the study on dynamic changes of MS lesions.

Objective To assess the changes of white matter microstructure in neuromyelitis optica spectrum disorders (NMOSD) with DTI based on tract-based spatial statistics (TBSS) method.Methods Conventional MR and DTI were performed in 20 NMOSD patients (NMOSD group) and 20 healthy volunteers (control group).DTI data were analyzed with TBSS procedure,which was a part of FSL software packages,and non-parametric statistical analysis was performed on the whole brain.Correlation between FA value of tracts with significant difference in NMOSD group and expanded disability status scale (EDSS) scores was analyzed.Results TBSS analysis revealed significantly (P＜ 0.05,FWE corrected) extensive decrease of FA value in cerebrum and cerebellum white matter fiber bundles,i.e.corpus callosum,fornix,corticospinal tract,unciform fasciculus,cerebellar peduncles etc.in NMOSD group,and decrease of AD value,increase of RD value in cingulum bundle,corpus callosum and fornix,while MD value only increased in fornix and retrolenticular part of the left internal capsule (P＜0.05,FWE corrected).Negative correlations were found between FA value of left uncinate fasciculus,right external capsule,left inferior cerebellar peduncle,bilateral anterior limb of internal capsule,medial lemniscus and EDSS scores (all P＜0.05).Conclusion Widespread white matter damage is observed in cerebrum and cerebellum in NMOSD patients by using TBSS analysis,which may partly correspond to the disabilities of NMOSD patients.However,the other microstructural changes of white matter tracts may suggest complicated pathological mechanism of NMOSD.

Objective To investigate the distribution of ideal cardiovascular health status in Chinese health checkup population. Methods Subjects were enrolled from a health checkup population coming to the PLA health management center from Sept 2009 to Mar 2016. Modified with China's specifications, lifestyle and checkup data were collected and analyzed according to the American ideal cardiovascular health standard. Results A total of 37664 people were included in the study, of whom 72.88%were male and 27.12%were female. Comprehensive analysis showed that, among 7 health indicators including smoking, physical activity, diet, fasting blood glucose, blood pressure, total cholesterol and body mass index, there were only 43 subjects (0.11%) whose lifestyle reached the ideal cardiovascular health status, 11 subjects were in the poor cardiovascular health status, accounting for 0.03%. The rest of the subjects were in the intermediate levels of cardiovascular health status. There was a large gap between the daily diet intake and the dietary recommendation, and there was also a large gap between the actual level physical activity and the ideal level of physical activity recommended by related guidelines, indicating that unhealthy diet and inadequate physical activity are two bottleneck factors. Dairy product intake has the lowest satisfaction ratio, followed by vegetable and fruit intake. Most subjects (94.10%) showed insufficient physical activity. The percentage of three status of cardiovascular health among young, middle-aged and elderly subjects differed significantly (χ2=1200, P=0.000), and presented an increasing trend of ideal cardiovascular health and a declining trend of poor cardiovascular health status with age from youth to middle age, to the elderly, which reflected insufficient physical activity especially among the young people, and then the middle-aged. Meanwhile, the proportion of ideal cardiovascular health in men was higher than that in women. Conclusion The rate of ideal cardiovascular health is relatively low in the study population, unhealthy diet and inadequate physical activity are two bottleneck factors. Encouraging people to develop good eating and exercise habits might be the most effective method to improve population's ideal cardiovascular health status.

Objective To assess the damage of limbic system pathways in relapsing-remitting multiple sclerosis (RRMS) by diffusion tensor imaging (DTI) tractography.Methods DTI tractography was used to acquire fiber numbers,fractional anisotropy (FA),mean diffusivity (MD) of cingulum,fornix,and uncinate fasciculus (UF) in 20 RRMS patients (MS group) and 20 healthy volunteers (control group),and statistical analysis was performed.Results Compared with control group,lower FA value (P＜0.01) and higher MD value (P＜0.05) were found in cingulum,fornix and UF of MS group,and the fiber numbers of fornix decreased in MS group (P＜0.001).There were significant differences between the left and the right UF in fiber numbers and FA value of control group and the fiber numbers of MS group.Negative correlations were found between FA value of cingulum and UF and EDSS scores in MS group (r=-0.572,-0.665,both P＜0.05),and positive correlations were found between MD value and EDSS scores (r=0.627,0.603,both P＜0.05).Conclusion DTI tractography is valuable in assessing MS associated limbic system damage and in monitoring the clinical progression of the disease.

Purpose Intravoxel incoherent motion (IVIM) MR is a method developed in recent years which can quantitatively evaluate the diffusion and perfusion characteristics of microenvironment.The aim of this study was to investigate the application value of IVIM in assessing relapsing-remitting multiple sclerosis (RRMS).Materials and Methods A retrospective analysis of the clinical data of 27 patients with RRMS confirmed clinically at the First Affiliated Hospital of Chongqing Medical University from Jun.2015 to Jan.2016 was carried out in the study.All the patients underwent the conventional MRI and IVIM MRI based on multi-b-factor (b values of 10,20,30,40,50,100,150,200,350,500,650,800,1000 s/mm2) with 3.0T MR scanner.The apparent diffusion coefficient (ADC),ADCslow,ADCfast and f values were evaluated since they could reflect the diffusion and perfusion status of RRMS lesions and normal-appearing white matter (NAWM) regions.Results The ADC,ADCslow,ADCfast and f values of the non-enhancing (NE) lesions were significantly higher than those in the NAWM regions both near and far from NE lesions (P＜0.05).However,the ADC,ADCslow,ADCfast and f values in the NAWM regions close to the NE lesions had no significant differences with those in the NAWM regions far from the lesions (P＞0.05).Conclusion The IVIM MRI can measure the diffusion and perfusion status of the lesions and NAWM in RRMS patients,which,therefore,is helpful in speculation of the pathological changes of RRMS lesions and in its injury classification and identification.

Objective To investigate the effects of different CO2 pneumoperitoneum pressures on gastric cancer cells' growth and proliferation in nude mouse model of implanted tumor. Methods Human gastric cancer cell lines MNK-45 were exposed under 0、10、12 and 15 mm Hg CO2 pneumoperitoneum for 4 hrs respectively. 2 × 106 processed cells were inplanted into nude mice subcutaneously. Three weeks later, mice were sacrificed and the weight and bulk of the tumor measured. Then we observed the transplantation tumor by HE stain and Ki-67 stain. Results There was no significant difference in tumor's growing time, bulk and weight between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (7. 8 d, 7. 2 d, 7. 8 d; 1. 2 cm3, 1. 3 cm3, 1. 3 cm3; 1.5 g, 1. 9 g, 1. 6 g)and the control group (7. 3 d, 1. 2 cm3, 1.4 g) (P > 0. 05 ). The growing time of tumor in 15 mm Hg CO2 pneumoperitoneum (12. 5 d) was obviously longer than the control group ( P < 0.05 ) , the bulk and weight of tumor in 15 mm Hg CO2 pneumoperitoneum (0. 5 cm3, 0. 5 g) group significantly decreased compared with the control group (P <0.05). The positive rate of Ki-67 in 15 mm Hg CO2 pneumoperitoneum (27. 5% ) group was obviously lower than the control group (59.6%) (P<0.01). However, there were no significant differences between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (61.2%, 60.5%, 63.4%) and the control group (P > 0.05). Conclusion Clinically adopted CO2 pneumoperitoneum pressures have no significant effect on gastric cancer cells growth and proliferation.

With the development of instrumentation and surgical techniques,laparoscopic gastrectomy has become a promising surgical option for the treatment of gastric cancer.While laparoscopic gastrectomy is high technique-demanding,which hampered its popularization.Compared with traditional laparoscopes,Da Vinci surgical system has more special features,such as flexible robotic arms and three-dimensional imaging,which facilitates surgical procedures.A 58-year-old male patient with gastric cancer underwent Da Vinci surgical system-assisted radical total gastrectomy at the Southwest Hospital in March 2010.The mean operation time and blood loss were 270 minutes and 60 ml,respectively,and the number of dissected lymph nodes was 21.The short-term clinical effect was perfect without postoperative complications.Da Vinci surgical system-assisted radical total gastrectomy is safe and feasible,and it brings challenges to conventional laparoscopes.

Objective To investigate the feasibility of laparoscopic-assisted transanal pull-through resection and anastomosis in the treatment of ultra-low rectal cancer.Methods From November 2005 to December 2006,21 patients with ultra-low rectal cancer had undergone laparoscopic-assisted transanal pull-through resection and anastomosis in Southwest Hospital.The perioperative condition,postoperative complications and the result of follow-up were retrospectively analyzed.Results The operation was successfully performed on all the patients.The mean operation time and postoperative hospital stay were(216±25)minutes(170-260 minutes)and(9.4±1.0)days(7-11 days),respectively.The time needed for the recovery of gastrointestina]function was(65±14)hours(38-88 hours).The mean perioperative blood loss was(140±49)ml(80-250 ml).All the patients were followed up for(22±4)months(15-28 months),and no anastomotic bleeding or fistula was observed.Six patients developed mild to moderate anastomotic striclure,1 local recurrence and 1 liver metastasis.Conclusions Laparoscopic-assisted transanal pull-through resection and anastomosis for ultra-low rectal cancer is safe and feasible,and the short-term effect is satisfactory.