Objective To explore the mechanism of the thioredoxin-interacting protein(TXNIP)/thioredoxin-1(Trx-1)pathway in regulating the polarization of retinal microglia in rats after retinal ischemia-reperfusion(RIR)in rats,and to provide new ideas for the prevention and treatment of retinal ischemia reperfusion injury(RIRI).Methods For-ty-two healthy adult male Sprague-Dawley rats were randomly divided into a Sham group,a RIRI group and a TXNIP siRNA group.The right eye of the rats was experimented.For RIRI and TXNIP siRNA groups,RIRI models were established using the anterior chamber high intraocular pressure method.Rats in the TXNIP siRNA group were given the intravitreal injection of TXNIP siRNA 3 d before modeling.Hematoxylin-eosin(HE)staining was used to analyze retinal histopathologic changes of rats in all groups 24 h after modeling.Immunohistochemical staining of brain-specific homeobox/POU domain proteins 3A(Brn-3a)was made to count the number of retinal ganglion cells(RGCs).The dynamical changes in the number of TXNIP+cells 6 h,24 h,72 h and 7 d after modelling were analyzed through immunohistochemical staining in the RIRI group.The retinal microglia polarization and changes in the expression of TXNIP and Trx-1 proteins in each group were de-tected by double immunofluorescence staining and Western blot 24 h after modeling.Results HE staining results showed that 24 h after modelling,the retinal cells were disordered and the inner retinal layer was thickened and swelled in RIRI and TXNIP siRNA groups,compared with those in the Sham group(all P＜0.05).Immunohistochemical staining results of Brn-3a showed that 24 h after modeling,the number of Brn-3a+cells in RIRI and TXNIP siRNA groups significantly decreased,compared with that in the Sham group(both P＜0.05).The number of Brn-3a+cells in the TXNIP siRNA group was signifi-cantly higher than that in the RIRI group(P＜0.05).Immunohistochemical staining results of TXNIP at different time points after modeling showed that the expression of TXNIP+proteins started to increase 6 h after modeling.The TXNIP+protein level reached a peak at 24 h and then decreased gradually.Western blot results revealed that 24 h after modeling,RIRI and TXNIP siRNA groups had significantly higher TXNIP levels and significantly lower Trx-1 levels than the Sham group(all P＜0.05).Compared with those in the RIRI group,the expression of TXNIP proteins was significantly lower and the expression of Trx-1 proteins was significantly higher in the TXNIP siRNA group(both P＜0.05).Double immunofluores-cence staining showed that 24 h after modeling,Iba1+/CD206+cells were significantly more and Iba1+/CD16+cells were significantly less in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).RIRI and TXNIP siRNA groups had significantly more Ibal+/TXNIP+cells and significantly less Iba1+/Trx-1+cells than the Sham group(both P＜0.05).The number of Iba1+/TXNIP+cells was significantly lower and the number of Iba1+/Trx-1+cells was significantly higher in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).Conclusion RIR activates the TXNIP/Trx-1 path-way to induce the activation of retinal microglia and regulate the polarization of microglia,thereby resulting in RIRI in rats.

Objective To explore the mechanism of the thioredoxin-interacting protein(TXNIP)/thioredoxin-1(Trx-1)pathway in regulating the polarization of retinal microglia in rats after retinal ischemia-reperfusion(RIR)in rats,and to provide new ideas for the prevention and treatment of retinal ischemia reperfusion injury(RIRI).Methods For-ty-two healthy adult male Sprague-Dawley rats were randomly divided into a Sham group,a RIRI group and a TXNIP siRNA group.The right eye of the rats was experimented.For RIRI and TXNIP siRNA groups,RIRI models were established using the anterior chamber high intraocular pressure method.Rats in the TXNIP siRNA group were given the intravitreal injection of TXNIP siRNA 3 d before modeling.Hematoxylin-eosin(HE)staining was used to analyze retinal histopathologic changes of rats in all groups 24 h after modeling.Immunohistochemical staining of brain-specific homeobox/POU domain proteins 3A(Brn-3a)was made to count the number of retinal ganglion cells(RGCs).The dynamical changes in the number of TXNIP+cells 6 h,24 h,72 h and 7 d after modelling were analyzed through immunohistochemical staining in the RIRI group.The retinal microglia polarization and changes in the expression of TXNIP and Trx-1 proteins in each group were de-tected by double immunofluorescence staining and Western blot 24 h after modeling.Results HE staining results showed that 24 h after modelling,the retinal cells were disordered and the inner retinal layer was thickened and swelled in RIRI and TXNIP siRNA groups,compared with those in the Sham group(all P＜0.05).Immunohistochemical staining results of Brn-3a showed that 24 h after modeling,the number of Brn-3a+cells in RIRI and TXNIP siRNA groups significantly decreased,compared with that in the Sham group(both P＜0.05).The number of Brn-3a+cells in the TXNIP siRNA group was signifi-cantly higher than that in the RIRI group(P＜0.05).Immunohistochemical staining results of TXNIP at different time points after modeling showed that the expression of TXNIP+proteins started to increase 6 h after modeling.The TXNIP+protein level reached a peak at 24 h and then decreased gradually.Western blot results revealed that 24 h after modeling,RIRI and TXNIP siRNA groups had significantly higher TXNIP levels and significantly lower Trx-1 levels than the Sham group(all P＜0.05).Compared with those in the RIRI group,the expression of TXNIP proteins was significantly lower and the expression of Trx-1 proteins was significantly higher in the TXNIP siRNA group(both P＜0.05).Double immunofluores-cence staining showed that 24 h after modeling,Iba1+/CD206+cells were significantly more and Iba1+/CD16+cells were significantly less in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).RIRI and TXNIP siRNA groups had significantly more Ibal+/TXNIP+cells and significantly less Iba1+/Trx-1+cells than the Sham group(both P＜0.05).The number of Iba1+/TXNIP+cells was significantly lower and the number of Iba1+/Trx-1+cells was significantly higher in the TXNIP siRNA group than those in the RIRI group(both P＜0.05).Conclusion RIR activates the TXNIP/Trx-1 path-way to induce the activation of retinal microglia and regulate the polarization of microglia,thereby resulting in RIRI in rats.

Objective:To investigate the effect of N-acetylserotonin (NAS) on the retinal microglia polarization in retinal ischemia-reperfusion injury (RIRI) rats and explore its mechanism via nucleotide-bound oligomeric domain 1 (NOD1)/receptor interacting protein 2 (Rip2) pathway.Methods:Healthy male Sprague Dawley rats were randomly divided into Sham ( n=21), RIRI ( n=21) and NAS (injected intraperitoneally 30 min before and after modeling with NAS, 10 mg/kg, n=18) groups, using random number table. And the right eye was used experimental eye. The RIRI model of rats in RIRI group and NAS group was established by anterior chamber high intraocular pressure method. Rats in NAS group were intraperitoneally injected with 10 mg/kg NAS before and 30 min after modeling, respectively. The retinal morphology and the number of retinal ganglion cell (RGC) in each group were detected by hematoxylin-eosin staining and immunohistochemical staining. The effect of NAS on polarization of retinal microglia was detected by immunofluorescence staining. Transcriptome sequencing technology was used to screen out the differentially expressed genes between Sham and RIRI groups. Western blot and real-time quantitative polymerase chain reaction (RT-PCR) were used to examine the differentially expressed genes. Immunohistochemical staining, Western blot and RT-PCR were used to investigate the effect of NAS on the expression of NOD1 and Rip2 protein and mRNA in retinal tissue and microglia of rats. General linear regression analysis was performed to determine the correlation between the number difference of NOD1 + cells and the number difference of M1 and M2 microglia in retinal tissues of rats in NAS group and RIRI group. Results:A large number of RGC were observed in the retina of rats in Sham group. 24 h after modeling, compared with Sham group, the inner retinal thickness of rats in RIRI group was significantly increased and the number of RGC was significantly decreased. The thickness of inner retina in NAS group was significantly thinner and the number of RGC was significantly increased. Compared with Sham group, the number of retinal microglia of M1 and M2 in RIRI group was significantly increased. Compared with RIRI group, the number of M1 microglia decreased significantly and the number of M2 microglia increased significantly in NAS group. There was statistical significance in the number of M1 and M2 microglia in the retina of the three groups ( P<0.05). Transcriptome sequencing results showed that retinal NOD1 and Rip2 were important differential genes 24 h after modeling. The mRNA and protein relative expressions of NOD1 and Rip2 in retina of RIRI group were significantly higher than those of Sham group, with statistical significance ( P<0.05). The number of NOD1 + and Rip2 + cells and the relative expression of mRNA and protein in retinal microglia in RIRI group were significantly higher than those in Sham group, and NAS group was also significantly higher than that in Sham group, but lower than that in RIRI group, with statistical significance ( P<0.05). The number of Iba-1 +/NOD1 + and Iba-1 +/Rip2 + cells in retinal microglia in RIRI group was significantly increased compared with that in Sham group, and the number of Iba-1 +/Rip2 + cells in NAS group was significantly decreased compared with that in RIRI group, but still significantly higher than that in Sham group, with statistical significance ( P<0.05). Correlation analysis results showed that the difference of retinal NOD1 + and Rip2 + cells in NAS group and RIRI group was positively correlated with that of M1 microglia ( r=0.851, 0.895), and negatively correlated with that of M2 microglia ( r=-0.797, -0.819). The differences were statistically significant ( P<0.05). Conclusion:NAS can regulate the microglial polarization from M1 to M2 phenotype, the mechanism is correlated with the NOD1/Rip2 pathway.

Objective To explore the effects of hyperbaric oxygen (HBO) on Th17/Treg cell infiltration in the brain tissue of Alzheimer disease (AD) mice,and also to identify possible mechanism involved for the improvement of AD conditions.Methods Following HBO treatment of APP/PS1 double transgenic mice,immunofluorescence staining and Western blotting were used to detect the deposition of Aβ in the brain tissue.Flow cytometry was used to analyze the infiltration of Th17 cells and Treg cells in the brain tissue.Results The deposition of Aβ in the brain tissue of AD mice after HBO therapy was significantly reduced (P ＜ 0.01).In addition,HBO would increase the expression of IL-17,while it could significantly decrease the expression of FOXP3 (P ＜ 0.01).Flow cytometry detection results indicated that HBO therapy could up-regulate the proportion of Th17/Treg cells in the brain tissue of AD mice (P ＜ 0.01).Conclusion HBO could decrease Aβ deposition in the brain tissue through up-regulation of the proportions of Thl7/Treg cells in the brain tissue of AD mice.

Objective To explore the effects of hyperbaric oxygen (HBO) on Th17/Treg cell infiltration in the brain tissue of Alzheimer disease (AD) mice,and also to identify possible mechanism involved for the improvement of AD conditions.Methods Following HBO treatment of APP/PS1 double transgenic mice,immunofluorescence staining and Western blotting were used to detect the deposition of Aβ in the brain tissue.Flow cytometry was used to analyze the infiltration of Th17 cells and Treg cells in the brain tissue.Results The deposition of Aβ in the brain tissue of AD mice after HBO therapy was significantly reduced (P ＜ 0.01).In addition,HBO would increase the expression of IL-17,while it could significantly decrease the expression of FOXP3 (P ＜ 0.01).Flow cytometry detection results indicated that HBO therapy could up-regulate the proportion of Th17/Treg cells in the brain tissue of AD mice (P ＜ 0.01).Conclusion HBO could decrease Aβ deposition in the brain tissue through up-regulation of the proportions of Thl7/Treg cells in the brain tissue of AD mice.

Objective To report a newly developed method and procedure to establish a rat model of subarachnoid hemorrhage in detail, and to provide a better model simulating the clinical subarachnoid hemorrhage caused by a ruptured aneurysm for related research.Methods One hundred and twenty healthy SPF 2-3-month old male Sprague-Dawley rats were divided into 4 groups, 30 rats in each group.The three experimental groups were sacrificed at 6, 24 and 72 hours after modeling.Rat models of subarachnoid hemorrhage were established by inserting a fiber core in the internal carotid artery and piercing this artery.Successful establishment of the subarachnoid hemorrhage model was confirmed by observation of breathing, pupil, defecation, urination and inspection at autopsy dissection.The controllability and reproducibility of this model were verified by observation of clinical manifestation and explored by mortality analysis.Results Subarachnoid hemorrhage was successfully induced by fiber core piercing the internal carotid artery at the needed location.Conclusions This method of model preparation is stable and understandable.The operation is nimble, with a good reproducibility.This model can be successfully performed after a short time learning, well simulate the sudden hemorrhage caused by a ruptured aneurysm, and suitable for research on early brain injury and vasospasm after subarachnoid hemorrhage.

Objective:To characterize the clinical and epidemiological changes, treatments, and prognoses of primary esophageal small cell carcinoma (PESC). Methods:A retrospective analysis was conducted using the clinical epidemiology data of 529 PESC patients se-lected from the clinical databases of 500,000 esophageal and gastric cardiac carcinomas of the Henan Key Laboratory for Esophageal Cancer Research (1992-2015). Among these patients, 241 cases were included in the survival analysis. The five-year survival rate was calculated using Kaplan-Meier analysis, and the differences in survival rates were compared using the Log-rank analysis model. Re-sults:All 529 PESC cases were analyzed, which accounted for 0.2%of esophageal cancers diagnosed in the same period. The incidence of PESC increased annually (R2=0.574). The survival rates for 1-, 2-, 3-, and 5-year of 241 PESC patients were 55%, 40%, 29%, and 9%, respectively, and the median survival time was 21.9 months. On the basis of the VALSG criteria of lung small cell carcinoma, a statisti-cal difference was observed in the overall survival rates for limited and extensive diseases (P=0.003), with the median survival time of 24.3 and 17.5 months, respectively. Furthermore, significant differences were observed on survival with various treatment modalities (P=0.004). The median survival time of PESC patients treated with combined surgery and radiochemotherapies (28.8 months) was lon-ger than those with either chemotherapy (17.8 months, P=0.015) or chemoradiotherapy (14.5 months, P=0.004). In limited disease pa-tients, the median survival time was longer in patients treated with surgery (27.7 months) than in those without surgery (16.2 months, P=0.007). Notably, the biopsy diagnosis before surgery for PESC was only 40.8%. Conclusion:PESC is a rare malignant carcinoma with increasing incidence. PESC presents poor prognosis, and the survival rate can be improved through combined therapies based on sur-gery. A high misdiagnosis rate for PESC is observed before surgery with biopsy.

Objective To explore the effect of hyperbaric oxygen (HBO) on the anxiety and depression after traumatic brain injury (TBI) and optimal treatment course.Methods One hundred and five TBI patients with depression or anxiety,who either sought medical care in the outpatient or hospitalized were recruited 干or the study.Of all the cases,56 were diagnosed to have anxiety,38 have depression,and another 11 cases were diagnosed to have both anxiety and depression.The patients were divided into the HBO group and the non-HBO group,depending upon whether or not they received HBO therapy.The patients in the HBO group were those treated with HBO in our department for at least 4 weeks,while the patients in the non-HBO group were those who received medical treatment,but without HBO therapy or those who received HBO therapy for less than 1 week.Of the 105 patients,59 received HBO therapy and 46 did not have HBO therapy.In the 67 patients with anxiety,38 received HBO therapy,while 29 did not have HBO therapy.On the other hand,in the 49 patients with depression,28 received HBO therapy,while 21 did not have HBO therapy.In the case of patients with anxiety,the scores of self-rating anxiety scale (SAS) and Hamilton anxiety scale (HAMA),1,2,3 and 4 weeks after treatment were assessed and compared between the 2 groups.And in the case of patients with depression,the scores of self-rating depression scale (SDS) and Hamilton depression scale (HAMD),1,2,3 and 4 weeks after treatment were also assessed and compared between the 2 groups.Results For all the patients,the SAS and SDS scores of the HBO group(44.9 ± 11.2,36.8 ± 14.0)4 weeks after therapy were significantly lower than those of the non-HBO group(5 1.7 ± 13.9,42.8 ± 14.9)at the same time point (P ＜0.05).For the patients with anxiety,no significant differences could be noted in the scores of SAS and HAMA,1 and 2 weeks after HBO therapy,as compared with those of the non-HBO group (P ＞ 0.05).However,the scores of SAS and HAMA,3 and 4 weeks after HBO therapy were significantly decreased,as compared with those of either before therapy or 1 and 2 weeks after HBO therapy.Furthermore,the scores of the HBO group were significantly lower than those of the non-HBO group,with statistical significance (P ＞0.05).For the patients with depression,no significant differences could be noted in the SAS and HAMA scores in the patients of the HBO group following 1 and 2 weeks after HBO therapy,when compared with those of the non-HBO group(P ＞ 0.05).However,the scores of SAS and HAMA 4 weeks after HBO therapy were significantly decreased,as compared with those 3 weeks after HBO therapy,and was significantly lower than those of the non-HBO group at the same time point,and statistical significance could be seen when comparison were made between the two(P ＞ 0.05).Conclusions HBO therapy could obviously alleviate the symptoms of the patients with anxiety and depression.However,an over 3-week treatment course was required for the patients with anxiety,and an over 4-week treatment course was mandatory for those patients with depression.

Objective To explore the effect of hyperbaric oxygen (HBO) on the anxiety and depression after traumatic brain injury (TBI) and optimal treatment course.Methods One hundred and five TBI patients with depression or anxiety,who either sought medical care in the outpatient or hospitalized were recruited 干or the study.Of all the cases,56 were diagnosed to have anxiety,38 have depression,and another 11 cases were diagnosed to have both anxiety and depression.The patients were divided into the HBO group and the non-HBO group,depending upon whether or not they received HBO therapy.The patients in the HBO group were those treated with HBO in our department for at least 4 weeks,while the patients in the non-HBO group were those who received medical treatment,but without HBO therapy or those who received HBO therapy for less than 1 week.Of the 105 patients,59 received HBO therapy and 46 did not have HBO therapy.In the 67 patients with anxiety,38 received HBO therapy,while 29 did not have HBO therapy.On the other hand,in the 49 patients with depression,28 received HBO therapy,while 21 did not have HBO therapy.In the case of patients with anxiety,the scores of self-rating anxiety scale (SAS) and Hamilton anxiety scale (HAMA),1,2,3 and 4 weeks after treatment were assessed and compared between the 2 groups.And in the case of patients with depression,the scores of self-rating depression scale (SDS) and Hamilton depression scale (HAMD),1,2,3 and 4 weeks after treatment were also assessed and compared between the 2 groups.Results For all the patients,the SAS and SDS scores of the HBO group(44.9 ± 11.2,36.8 ± 14.0)4 weeks after therapy were significantly lower than those of the non-HBO group(5 1.7 ± 13.9,42.8 ± 14.9)at the same time point (P ＜0.05).For the patients with anxiety,no significant differences could be noted in the scores of SAS and HAMA,1 and 2 weeks after HBO therapy,as compared with those of the non-HBO group (P ＞ 0.05).However,the scores of SAS and HAMA,3 and 4 weeks after HBO therapy were significantly decreased,as compared with those of either before therapy or 1 and 2 weeks after HBO therapy.Furthermore,the scores of the HBO group were significantly lower than those of the non-HBO group,with statistical significance (P ＞0.05).For the patients with depression,no significant differences could be noted in the SAS and HAMA scores in the patients of the HBO group following 1 and 2 weeks after HBO therapy,when compared with those of the non-HBO group(P ＞ 0.05).However,the scores of SAS and HAMA 4 weeks after HBO therapy were significantly decreased,as compared with those 3 weeks after HBO therapy,and was significantly lower than those of the non-HBO group at the same time point,and statistical significance could be seen when comparison were made between the two(P ＞ 0.05).Conclusions HBO therapy could obviously alleviate the symptoms of the patients with anxiety and depression.However,an over 3-week treatment course was required for the patients with anxiety,and an over 4-week treatment course was mandatory for those patients with depression.

Objective To investigate the characteristics of fractional anisotropy (FA) and apparent diffusion coefficient (ADC) of association fiber tracts in patients with amnestic mild cognitive impairment (aMCI), and to assess the application value of diffusion tensor imaging (DTI) in differential diagnosis of aMCI and AD. Methods DTI were performed in 20 patients of aMCI (aMCI group), 20 patients of AD (AD group) and 20 normal control subjects (control group). FA and ADC values were calculated in the regions of interest (ROI) in inferior fronto-occipital fasciculus (IFOF), genu and splenium of corpus callosum, superior longitudinal fasciclesⅡ (SLFⅡ) and cingulated bundles. Results There was significant difference of FA values in inferior fronto-occipital fasciculus and cingulate bundles between aMCI group compared with control group (P<0.05), as well as of FA values in cingulate bundles between aMCI group and AD group (P<0.05). Conclusion Abnormal FA values in inferior fronto-occipital fasciculus and cingulate bundles suggest that DTI can be used as a diagnosis index of aMCI. Furthermore, it is helpful in the differential diagnosis of aMCI and AD.