A model of s bacute senility state was established in mice by successive retrobulbar injectiou of 80 mg/kg D-galactose for 50 days. Concomitant oral administration of 4 g/kg and 8 g/kg gastrodia can effectively recovcr the decreased ability of the passive avoid ance reaction in the model animals.The activity of RBC SOD and the contents of hydroxyproline in the animals skin were markedly increased as well as the lipofuscin of myocardium. However, gastrodia was less effective to lower lipofuscin in liver and brain.

0.05).The death rate of cases with pneumonia combined with ascites was higher than that of cases with ascites only(?2=4.894,P=0.027) and cases without ascites and infections(?2=9.260,P=0.002).Unfavorable prognosis was found in cases with Enterococcus faecium isolated before OLT.CONCLUSIONS Severe lung infection before OLT is one of the main reasons of death.It is important to grasp characteristics of infection,evaluate risk fully,control infections and screen cases strictly before OLT to improve survival rate.

OBJECTIVE To investigate the value of fibrobronchoscopy and bronchoalveolar lavage in etiologic diagnosis of pneumonia in immunocompromised patients.METHODS The clinical document and results of fibrobronchoscopy and bronchoalveolar lavage in 36 immunocompromised patients with pneumonia were retrospectively analyzed,whose conditions were mainly after organ transplantation and hematologic neoplasia.RESULTS Through fibrobronchoscopy and(or) bronchoalveolar lavage,22 cases(61.1%) were etiologically diagnosed.In 19 cases taking cytomegalovirus(CMV) quantitative PCR test of both peripheral blood and BALF,the positive rate of blood and BALF was 14.3% and 42.9%,respectively(P

Objective To explore the influence of different dose of Helicobacter pylori on the expression of transforming growth factorβ(TGF-β1) and B7-H1 in the infected gastric mucosal epithelial cell and the bacterial factors which influence the expression of TGF-β1.To confirm that H.pylorican induce the expression of TGF-β1 and B7-H1 to inhibit the host immune function in gastric mucosal epithelial cell.Methods (1) We investigated the expression of TGF-β1 of human gastric mucosal epithelial cells infected with different concentration(1.0 × 109 CFU/ml,4.0 × 109 CFU/ml,8.0 × 109 C FU/ml) of H.pylori(NCTC 11637) in different time-point(0 h,0.5 h,1 h,1.5 h,2 h,4 h,8 h,12 h),and compared with the expression of TGF-β1 between the deactivated H.pylori group and activated H.pylori group.The blank group is the gastric mucosa epithelial cells which does not infect H.pylori.To detect expression of TGF-β1 in infected cell culture supernatant by enzyme-linked immunosorbent assay(ELISA) and the expression of B7-H1 mRNA by in situ hybridization.(2)At the same time,the middle concentration of deactivated H.pyloriand in vitro gastric mucosal cells were incubated for 2 h and 12 h,to detect expression of TGF-β1 in the cells and cell culture supernatant.(3)In vitro gastric mucosal cells were incubated with H.pylori bacterial supernatant and sedimentation by ultrasonic crushing and centrifugation and with H.pyloribacterial supernatant and sedimentation after boiling respectively,to detect expression of TGF-β1 in the cells and cell culture supernatant after 2 h,12 h.Results (1)Compared to the control group,the expression of TGF-β1 was significantly increased after stimulation with different concentration of activated H.pylori in different time-point(P ＜0.05).The expression of TGF-β1 secretion group has a similar dynamic trend,and the highest expression is the middle concentration group(P ＜0.05).(2)There was no difference between the middle concentration of deactivated H.pylori group and the same concentration of activated H.pylorigroup(P ＞ 0.05).(3) The expression of TGF-β1 in the H.pylori bacterial supernatant group was significantly increased higher than the blank group and the H.pylori bacterial sedimentation group(P ＜0.05),and the H.pylori bacterial supernatant group after boiling was significantly lower than the H.pylori bacterial supernatant group(P ＜ 0.05),but there was no difference between H.pylori sedimentation group after boiling and not boiling(P ＞ 0.05).The B7-H1 expression of different concentration groups which the H.pylori and gastric mucosal epithelial cells cocultured 12 h are higher than the blank group(P ＜ 0.05) by in situ hybridization,and the middle concentration group is the highest expression.TGF-β1 and B7-H1 mRNA are positively correlated(r,=0.628,P ＜0.01).Conclusion H.pylori can induce the gastric mucosal epithelial cells to express the TGF-β1,the factor was the soluble protein in the H.pylori thalline.At the same time,H.pylorican induce the B7-H1 expression increased.In gastric mucosal epithelial cells,TGF-β1 and B7-H1 mRNA are positively correlated.So H.pylori can inhibit the host immune response and participate the process of immune escape by increased the expressions of TGF-β1 and B7-H1.

Objective: To analyze the correlation and gender differences between adverse childhood experiences (ACEs) and positive childhood experiences (PCEs) and depression and anxiety symptoms among middle school students, so as to provide a reference for promoting the mental health of middle school students. Methods: With a stratified random cluster sampling method, a total of 6 656 middle school students in 4 cities, including Nanchang, Shenyang, Taiyuan, and Zhengzhou, were selected as research subjects from October 2021 to October 2022. The Adverse Childhood Experiences International Questionnaire (ACEs-IQ), Benevolent Childhood Experiences Scale (BCEs), Patient Health Questionnaire-9 (PHQ-9) and Generalized Anxiety Disorders-7 (GAD-7) scale were used to conduct questionnaire surveys.The Chi square test was used to compare the reporting rates of depression and anxiety symptoms among middle school students in different groups, and a Logistic regression model was established to analyze the effects of ACEs and PCEs on depression and anxiety symptoms among middle school students and their gender differences. Results: The reporting rate of depressive symptoms among middle school students was 20.1%, and the reporting rate of anxiety symptoms was 13.9% . ACEs were positively correlated with depression and anxiety symptoms among middle school students (depression symptoms: OR =1.20, 95% CI =1.18-1.22, anxiety symptoms: OR =1.18, 95% CI =1.16-1.20), while PCEs were negatively correlated with depression and anxiety symptoms among middle school students(depression symptoms: OR =0.84, 95% CI = 0.83 -0.86, anxiety symptoms: OR =0.85, 95% CI =0.83-0.87) ( P <0.05). In the general population (depression symptoms ： OR =0.99, 95% CI = 0.98- 0.99, anxiety symptoms: OR =0.99, 95% CI =0.99-1.00) and among girls (depression symptoms: OR = 0.98 , 95% CI = 0.97- 0.99 , anxiety symptoms ： OR =0.99, 95% CI =0.98-1.00), the interaction term between ACEs and PCEs were negatively correlated with depression and anxiety symptoms ( P <0.05). Conclusions ACEs significantly affect the depression and anxiety symptoms of middle school students, while PCEs can help reduce the impact of ACEs on the depression and anxiety symptoms of middle school students, girls are more susceptible to the impact of early experiences than boys. It should focus on gender differences, formulate comprehensive mental health protection strategies, to promote the mental health development of middle school students.

Objective To investigate the regulating effects of Krüppel-like factor 6 (KLF6) on the apoptosis of human lens epithelial cells (HLECs) by activating transcription factor 4 (ATF4) pathway and explore the bio-molecular mechanism of KLF6/ATF4-induced HLECs apoptosis.Methods HLECs (HLE-B3) were cultured using high glucose DMEM medium.The eukaryotic expression plasmid pEGFP-C2-ATF4 was transfected into the cells by liposome 2000 in the ATF4-transfected group,and pEGFP-C2 was transfected in the empty plasmid group.Then the cells were exposed to 20 mJ/cm2 ultraviolet ray B (UVB) for 200 seconds,The morphological changes of the cells were observed by hematoxylin & eosin staining and Hoechst33258 fluorescein staining.Cultured cells were transfected using pEGFP-C2-KLF6 and pEGFP-C2 plasmid and pSilencer-KLF6 (siKLF6) and pSilencer plasmid,respectively,and the expression of ATF4 protein in the cells was detected by Western blot assay.Culture cells were divided into four groups.pEGFP-C2 and pSilencer plasmids were co-transfected into the cells in the empty plasmid group;pEGFP-C2-KLF6 and pSilencer empty plasmid were co-transfected into the cells of the KLF6 + pSilencer group;pEGFP-C2 empty plasmid and pSilencer-ATF4 were co-transfected in the cells of the siATF4 + pEGFP-C2 group;pEGFP-C2-KLF6 and pSilencer-ATF4 plasmids were co-transfected in the cells of the KLF6 + siATF4 group,and then the cells were exposed to UVB.The apoptosis of the cells were detected by ELISA assay.Results Cultured cells grew well in the normal control group with the uniform morphology and regular arrangement.The karyopyknosis,karyorrhexis and enlargement of intercellular space were found in the cells exposed to UVB.In the ATF4 transfected group,the number of cells was decreased.The relative expression level of the ATF4 protein in the cells was 0.99±0.06 and 0.13±0.02 in the UVB+ATF4 transfected group and UVB+pEGFP-C2 plasmid group,respectively,with a significant difference between them (t =23.13,P＜0.01).The relative expression levels of KLF6 and ATF4 proteins in the KLF6 transfected group were higher than those in the empty plasmid group,and the relative expression levels of KLF6 and ATF4 proteins in the siKLF6 group were significantly lower than those in the empty plasmid group (all at P＜0.01).ELISA assay showed that the apoptotic rate in the ATF4 transfected group was 1.37± 0.11,which was significantly higher than 0.31 ±0.11 in the normal control group (t =8.034,P =0.001);the apoptotic rate of the cells was increased in the KLF6+pSilencer group and decreased in the siATF4+pEGFP-C2 group in comparison with the empty plasmid group (P＜0.01,P=0.02).In addition,the apoptotic rate in the KLF6+ siATF4 group was remarkably lower than that in the KLF6 + pSilencer group (P＜ 0.01).Conclusions KLF6 promotes the apoptosis of HLECs induced by UVB radiation.Silence of ATF4 gene reduces the apoptotic rate of the cells.ATF4 is probably a target factor in the regulating oathwav of KLF6 to apoptosis.

Objective To observe the protective effect of dl-3-n-Butylphthalide (NBP) on apoptosis of retinal Müller cells induced by hydrogen peroxide (H2O2).Methods Human retinal Müller cells cultured in vitro were divided into normal control group,model group (H2O2 group) and experimental group (H2O2+NBP group).The cells in the H2O2 group and H2O2+NBP group were cultured with 200 μ mol/L H2O2 for 2 h.Then the culture solution of the H2O2 group replace with complete medium and the H2O2+NBP group replace with complete medium containing 1 tmol/L NBP.The normal control group was a conventional cultured cells.Müller cells were identified by immunofluorescence staining.Hematoxylin-eosin (HE) staining was used to observe the apoptosis morphological changes.MTT assay was used to detect the activity of of retinal Müller cells after after 24 h and 48 h of NBP intervention.Hoechst33258 staining was used to observe the apoptosis.LIVE/DEAD (R)cell activity/cytotoxicity kit was used to detect cell viability.Dichlorofluorescein diacetate (DCFH-DA) + endoplasmic reticulum (ER) red fluorescent probe (ER-Tracker Red) double staining was used to observe the expression level of reactive oxygen species (ROS) in ER of cells.One-way ANOVA combined with Dunnett statistical method were used for data analysis.Results HE staining showed that the number of cells in H2O2+NBP group was higher than that in H2O2 group.MTT assay showed that after 24 h and 48 h of NBP intervention,the differences in cell viability between the normal control group and the H2O2 group,the H2O2 group and the H2O2+NBP group were statistically significant (t=28.96,3.658,47.58,20.33;P＜0.001,0.022).The results of Hoechst33258 showed that the nuclear nucleus of a few cells in the H2O2+NBP group was crescent-shaped and the nuclear fragmentation was reduced,and the blue fluorescence of the remaining cells was uniform.The LIVE/DEAD ~ cell activity/cytotoxicity kit showed that the number of dead cells with red fluorescence in the H2O2 group increased significantly,and the number of viable cells with green fluorescence decreased significantly.In the H2O2+NBP group,the number of viable cells with green fluorescence increased,and the number of dead cells with red fluorescence decreased.The double staining results of DCFH-DA+ER-Tracker Red showed that the green fluorescence intensity of H2O2 group was significantly enhanced;the green fluorescence intensity of H2O2+NBP group was lower than that of H2O2 group.Conclusion NBP alleviates H2O2-induced apoptosis of human retinal Müller cells by inhibiting ROS production.

Objective To explore the role of forkhead box F2 (FoxF2) in the extracellular matrix of trabecular meshwork.Methods The cultured human trabecular meshwork cells (HTMCs) were divided into Scramble control group and FoxF2 small hairpin RNA (shRNA) group,then FoxF2 shRNA,the FoxF2 restructuring interference carrier was built,HTMCs were infected with FoxF2 shRNA lentivirus.Western blot assay was used to detect the expression of FoxF2 protein and extracellular matrix.Furthermore,Transwell counting experiment was used to analyze the migration ability of HTMCs.Results The cultured HTMCs grew well and showed a long spindle shape.The growth status of HTMCs was well,and their morphological characteristics were consistent with the HTMCs in vivo.The relative expression level of FoxF2 protein in the FoxF2 shRNA group was lower than that in the Scramble control group,with a significant difference between them (0.72 ± 0.02 vs.1.27 ± 0.05;t =16.68,P ＜ 0.01).The relative expression level of fibronectin (FN),collagen type Ⅰ (COL Ⅰ) and α-smooth muscle actin (α-SMA) were 0.43±0.03,0.53 ±0.08 and O.86±0.15 in the FoxF2 shRNA group,and 0.87±0.04,1.66±0.06 and 1.73 ±0.13 in the Scramble control group,respectively,the relative expression levels of FN,COL Ⅰ and α-SMA in the FoxF2 shRNA group were significantly lower than those in the Scramble control group (t =15.08,18.81,7.50,all at P＜0.01).The migration number of HTMCs in the FoxF2 shRNA group was significantly lower than that in the Scramble control group (117.30±11.41 vs.251.00±10.37;t =8.72,P＜0.01).Conclusions The FoxF2 shRNA lentivirus are successfully constructed,which can decrease the expression of FoxF2 in HTMCs.Low expression of FoxF2 can reduce the expression level of extracellular matrix protein in HTMCs and inhibit the migration ability of HTMCs.

Objective To observe the effect ofpolypyramidine tract binding protein-associated splicing factor (PSF) towards advanced glycation end products (AGEs) induced the apoptosis of Müller cells in vitro.Methods Experimental study.Müller cells were cultured and divided into groups according to the project design,plasmid enhanced green fluorescent protein-PSF were transfected into the cells to achieve the overexpression of PSF Müller cells in vitro,then cells were exposed to AGEs and the Morphological changes were observed by HE staining and Hoechst 33258 staining while the survival rate of cells were detected by MTT assay.The effects of PSF on AGEs-induced Müller apoptosis was measured by Cell Death Detection ELISA kit.Meanwhile,2',7'-diehlorofluorescin diaeetate staining was performed to monitor the protective effects of PSF on AGEs-induced Müller cells ROS.Results The morphology of cells in normal group was full and the cytoplasm staining was uniform.In N+AGEs group and Vec+AGEs group,cell volume decreased,cytoplasm was dense and concentrated,and eosinophilic staining was enhanced.The cell morphology of PSF+AGEs group was still full,with uniform cytoplasm staining and uniform nucleus staining.The viability of N+AGEs group,Vec+AGEs group and PSF+AGEs group were 0.42±0.11,0.35±0.12 and 0.68±0.12.The apoptosis values were 1.08 ± 0.16,0.96± 0.20 and 0.44± 0.08.The intracellular ROS levels were 28 833.67± 3 550.06,28 356.67±4 854.81,186 163.00±382.54.Compared with N+AGEs group and Vec+AGEs group,the cell viability of PSF+AGEs group was significantly improved (F=20.65,P=0.000),ce11 apoptosis value (F=43.43,P=0.000) and intracellular ROS level (F=1 8.86,P=0.000).Conclusion PSF overexpression play a protective role in AGEs-induced apoptosis by inhibiting the production of ROS in Müiller cells.

OBJECTIVETo investigate the feasibility of using liquid chromatography (HPLC) to characterize the 3, 4-Dihydroxyphenylalanine (DOPA) redox state of mussel adhesive protein (MAP).

METHODSThe DOPA and protein contents of MAP were determined by HPLC, Arnow and Bradford methods respectively.

RESULTSWith extended oxidation time, the protein contents of MAP samples remained unchanged whereas the DOPA contents declined. The retention times of main peaks in HPLC for both the accelerated oxidation and retained samples shifted as the storage time extended, which could be related to the changes of sample redox state.

CONCLUSIONSThe redox state of MAP can be characterized by the change of HPLC peak retention time. HPLC can be used in the research on the MAP redox state, which is beneficial to the product development and quality control.