AIM:To investigate the protective effect and mechanism of zinc ions on lung ischemia/reperfusion injury(LIRI)in rats.METHODS:SPF SD rats aged 6～8 weeks were divided randomly into 4 groups:control(control)group,ischemia/reperfusion(I/R)group,zinc chloride(ZnCl2)+I/R group,and PI3K inhibitor(LY294002)+ZnCl2+I/R group.Inductively coupled plasma mass spectrometry(ICP-MS)was used to measure the concentration of zinc ions in lung tissues,while the degree of lung tissue injury was assessed by HE staining,the alveolar damage index,and the lung wet/dry weight ratio.qPCR was used to detect the mRNA expression of solute carrier family 39 member 8(SLC39A8/ZIP8),with the TUNEL assay used to determine the level of apoptosis in lung tissue.The phosphorylation levels of caspase3,PI3K,AKT,GSK-3β,ZIP8,and solute carrier family 30 member 9(SLC30A9/ZNT9)were detected by Western blot,while the mitochondrial membrane potential was measured by the mitochondrial extraction kit and JC-1 mitochondrial mem-brane potential detection kit.RESULTS:Compared with the I/R group,the zinc ion level in the ZnCl2+I/R group in-creased(P<0.01),with the qPCR results showing that the expression level of ZIP8 also increased(P<0.01).The West-ern blot results demonstrated that the phosphorylation levels of PI3K/AKT/GSK-3β and cleaved caspase-3/pro were both in-creased(P<0.01 or P<0.05).In addition,the level of caspase-3 was decreased(P<0.01),the ZIP8 level was increased(P<0.05),whereas the level of ZNT9 was not significantly different(P>0.05).The mitochondrial membrane potential was increased(P<0.01)and the level of apoptosis was decreased(P<0.01).The results of HE staining,total lung water(TLW),lung index of quantitative assessment of histology(IQA),and lung tissue wet/dry weight ratio showed that the de-gree of injury was reduced significantly(P<0.05 or P<0.01).Compared with the ZnCl2+I/R group,the LY294002+Zn-Cl2+I/R group had a significant reduction in zinc ion levels(P<0.05),while qPCR showed a significant reduction in ZIP8 expression(P<0.01).Western blot showed that the phosphorylation level of PI3K/AKT/GSK-3β was decreased(P<0.01),the level of caspase-3/pro-caspase-3 was increased(P<0.01)the level of ZIP8 was decreased(P<0.05),al-though there was no significant difference in ZNT9(P>0.05).Measurements of the mitochondrial membrane potential demonstrated a significant decrease(P<0.01),while the TUNEL results showed that the level of apoptosis had increased(P<0.05).HE staining,TLW,IQA and lung tissue wet/dry weight ratio indicated that the degree of injury was aggravated significantly(P<0.05 or P<0.01).CONCLUSION:Administration of zinc chloride in rats has a protective role in lung ischemia/reperfusion injury by activating the PI3K/AKT pathway,leading to inactivation of GSK-3β,stabilization of the mitochondrial membrane potential level,and inhibition of cell apoptosis.

AIM:To investigate the protective effect and mechanism of zinc ions on lung ischemia/reperfusion injury(LIRI)in rats.METHODS:SPF SD rats aged 6～8 weeks were divided randomly into 4 groups:control(control)group,ischemia/reperfusion(I/R)group,zinc chloride(ZnCl2)+I/R group,and PI3K inhibitor(LY294002)+ZnCl2+I/R group.Inductively coupled plasma mass spectrometry(ICP-MS)was used to measure the concentration of zinc ions in lung tissues,while the degree of lung tissue injury was assessed by HE staining,the alveolar damage index,and the lung wet/dry weight ratio.qPCR was used to detect the mRNA expression of solute carrier family 39 member 8(SLC39A8/ZIP8),with the TUNEL assay used to determine the level of apoptosis in lung tissue.The phosphorylation levels of caspase3,PI3K,AKT,GSK-3β,ZIP8,and solute carrier family 30 member 9(SLC30A9/ZNT9)were detected by Western blot,while the mitochondrial membrane potential was measured by the mitochondrial extraction kit and JC-1 mitochondrial mem-brane potential detection kit.RESULTS:Compared with the I/R group,the zinc ion level in the ZnCl2+I/R group in-creased(P<0.01),with the qPCR results showing that the expression level of ZIP8 also increased(P<0.01).The West-ern blot results demonstrated that the phosphorylation levels of PI3K/AKT/GSK-3β and cleaved caspase-3/pro were both in-creased(P<0.01 or P<0.05).In addition,the level of caspase-3 was decreased(P<0.01),the ZIP8 level was increased(P<0.05),whereas the level of ZNT9 was not significantly different(P>0.05).The mitochondrial membrane potential was increased(P<0.01)and the level of apoptosis was decreased(P<0.01).The results of HE staining,total lung water(TLW),lung index of quantitative assessment of histology(IQA),and lung tissue wet/dry weight ratio showed that the de-gree of injury was reduced significantly(P<0.05 or P<0.01).Compared with the ZnCl2+I/R group,the LY294002+Zn-Cl2+I/R group had a significant reduction in zinc ion levels(P<0.05),while qPCR showed a significant reduction in ZIP8 expression(P<0.01).Western blot showed that the phosphorylation level of PI3K/AKT/GSK-3β was decreased(P<0.01),the level of caspase-3/pro-caspase-3 was increased(P<0.01)the level of ZIP8 was decreased(P<0.05),al-though there was no significant difference in ZNT9(P>0.05).Measurements of the mitochondrial membrane potential demonstrated a significant decrease(P<0.01),while the TUNEL results showed that the level of apoptosis had increased(P<0.05).HE staining,TLW,IQA and lung tissue wet/dry weight ratio indicated that the degree of injury was aggravated significantly(P<0.05 or P<0.01).CONCLUSION:Administration of zinc chloride in rats has a protective role in lung ischemia/reperfusion injury by activating the PI3K/AKT pathway,leading to inactivation of GSK-3β,stabilization of the mitochondrial membrane potential level,and inhibition of cell apoptosis.

AIM:To investigate the effects of sodium hydrosulfide(NaHS)on hexokinase 2(HK2)-nucleo-tide-binding oligomerization domain-like receptor protein 3(NLRP3)-gasdermin D(GSDMD)pathway and pyroptosis in-duced by lung ischemia/reperfusion(I/R)in rats.METHODS:Male Sprague-Dawley rats were divided into 6 groups:control group,control+NaHS group,I/R group,low-dose NaHS+I/R(L+I/R)group,medium-dose NaHS+I/R(M+I/R)group,and high-dose NaHS+I/R(H+I/R)group,with 6 rats in each group.The NaHS was administered via intraperi-toneal injection at 1.5 mL,30 min before modeling.The left lung tissues were collected 30 min after ischemia and 1 h af-ter reperfusion,and the wet/dry weight ratio and total lung water content were recorded.Hematoxylin-eosin(HE)staining was used to examine lung tissue morphological changes.The levels of malondialdehyde(MDA),myeloperoxidase(MPO)and lactate in lung tissues were measured with test kits.ELISA was employed to determine the levels of interleukin-1β(IL-1β)and IL-18.The expression of glycolysis-and pyroptosis-related indicators was analyzed by Western blot,qRT-PCR and immunofluorescence staining.RESULTS:Compared with control group,the rats in NaHS group showed no signifi-cant differences in all laboratory tests(P>0.05).The rats in I/R group exhibited significant lung injury,oxidative stress,increased lactate level,and up-regulated glycolysis and pyroptosis(P<0.05 or P<0.01).Compared with I/R group,the indicators in L+I/R group showed a downward trend(P<0.01)or no difference(P>0.05),while those in M+I/R group dis-played a significant reduction(P<0.05 or P<0.01).However,the indexes in H+I/R group exhibited no significant dif-ferences in these tests(all P>0.05).CONCLUSION:A moderate dose(56 μmol·L-1·kg-1)of NaHS mitigated the oc-currence of pyroptosis by inhibiting the HK2-NLRP3-GSDMD pathway,thus contributing to the attenuation of lung I/R in-jury in rats.

Objective To explore the value of curvature value of liver surface nodularity(LSN)based on MRI in evaluating liver function in patients with liver cirrhosis.Methods A retrospective analysis was made on the patients who underwent upper abdomen MR examination at 3.0T.The normal liver function patients and cirrhosis patients were enrolled in the study and then the Child-Pugh score of the patients were calculated.The patients were divided into three groups:normal liver group,compensated cirrhosis group and decompensated cirrhosis group.The water phase imaging of 3D modified Dixon fast field echo(mDixon-FFE)sequence was copied in DICOM format.ITK software was used to manually draw the full-thickness liver edge by two observers.The curvature value of LSN was obtained by using matlab self compiled code for follow up analysis.Kruskal-Wallis H test was used to compare the curvature value between the groups.The receiver operating characteristic(ROC)curve was drawn and the area under the curve(AUC)was obtained.Spearman test was used for the correlation analysis.Results The curvature values of LSN among the normal liver,compensated cirrhosis and decompensated cirrhosis groups gradually increased(P<0.05).Comparing normal liver with compensated cirrhosis,the AUC of diagnosing compensated cirrhosis was 0.84,with the sensitivity of 72.7%and the specificity of 89.3%.Comparing compensated cirrhosis with decompensated cirrhosis,the AUC of diagnosing decompensated cirrhosis was 0.91,with the sensitivity of 80%and the specificity of 90.9%.There was a moderate positive correlation between the curvature value of LSN and liver function score in patients with cirrhosis(r=0.63,P=0.002).Conclusion The curvature value of LSN based on MRI can be used for preliminary evaluation of liver function of liver cirrhosis,with the AUC more than 0.80 and higher sensitivity and specificity.

AIM:To investigate the regulatory role of retinoid X receptor(RXR)in oxidative stress response of rat type Ⅱ alveolar epithelial cells(AECII)induced by hypoxia/reoxygenation(HR).METHODS:The AECII were di-vided into control(C)group,HR group,HR+solvent dimethyl sulfoxide(DMSO)group(HD group),HR+RXR agonist 9-cis-retinoic acid(9-RA)group(RA group),and HR+RXR antagonist HX531 group(HX group).Cell Counting Kit-8(CCK-8)method was used to measure the cell viability.Immunofluorescence staining was used to detect the expression of surfactant protein A(SP-A)and RXRα in AECII.Kits were detected to the levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in cells.Transmission electron microscopy was used to observe the ultrastructural changes of the cells.Western blot was used to detect the protein level of nuclear factor E2-related factor 2(Nrf2).RT-PCR was used to detect the expression level of Nrf2 mRNA.RESULTS:Compared with C group,the cell viability and SOD activity in HR,HD,RA and HX groups were decreased significantly(P<0.05),the MDA content were increased significantly(P<0.05),the Nrf2 mRNA and protein expression levels were decreased significantly(P<0.05 or P<0.01),and the immuno-fluorescence expression of RXRα was significantly increased(P<0.01).Compared with HR and HX groups,the cells in RA group showed significantly increased cell viability(P<0.05),increased SOD activity(P<0.05),decreased MDA con-tent(P<0.05),increased Nrf2 mRNA and protein expression levels(P<0.01),and significantly increased immunofluo-rescence expression of RXRα(P<0.01).CONCLUSION:Hypoxia/reoxygenation can aggravate the oxidative stress re-sponse of rat AECII,and RXR agonist intervention can alleviate HR-induced rat AECII injury by inhibiting oxidative stress.

Objective:To explore the β-amyloid (Aβ) deposition pattern of subjects with the preclinical Alzheimer′s disease (AD), community-derived amnestic mild cognitive impairment (aMCI) and normal cognition (NC) from communities of Shanghai.Methods:According to the inclusion and exclusion criteria, 273 subjects (104 males, 169 females; age (64.2±7.6) years) were recruited from Shanghai community and memory clinics from December 2018 to July 2020. All subjects underwent MRI, 18F-AV45 PET imaging and neuropsychological scale tests and were grouped into AD, aMCI and NC groups based on clinical diagnosis. Differences in demographic information, the neuropsychological scale tests′ scores and positive rate of Aβ deposition among each group were analyzed by one-way analysis of variance or χ2 test. Aβ deposition patterns of AD and MCI groups were analyzed at voxel level, and the differences of Aβ deposition among different groups were compared. Results:Among 273 patients, the positive rates of Aβ deposition in AD, aMCI and NC groups were 84.4%(38/45), 36.4%(20/55) and 23.1%(40/173), respectively ( χ2=58.37, P<0.001). Among AD, aMCI, NC and NC (Aβ-) groups ( n=132), the education years of AD group was the lowest ((9.7±4.6) years; F=8.86, P<0.001). In addition, there were significant differences in the scores of several neuropsychological scale tests among AD, aMCI, NC groups and NC (Aβ-) group ( F values: 27.68-235.50, all P<0.001). Compared with subjects in NC(Aβ-) group, the Aβ depositions in the aMCI and AD groups were widely distributed in the whole cerebral cortex; and AD group had higher Aβ deposition in bilateral frontal, parietal, temporal, occipital lobe, cingulate gyrus and precuneus than aMCI group. Conclusions:The positive rate of Aβ deposition in the preclinical AD population from the Shanghai community is obtained. There are significant different Aβ deposition patterns in subjects at different stages of AD.

Objective To identify abnormal cerebral glucose metabolism characteristics in patients with corticobasal degeneration (CBD) using 18 F-fluorodeoxyglucose (FDG) PET imaging. Methods From January 2014 to January 2017, resting-state brain 18 F-FDG PET imaging was performed in 10 CBD patients (5 males, 5 females; average age: (63.4±6.2) years) and 20 age-matched healthy subjects (8 males, 12 female; average age: (63.6±6.2) years). Voxel-based statistical parametric mapping (SPM) was used to analyze images to obtain the CBD-related brain metabolic characteristics. The regional cerebral metabolic rate of glucose (rCMRglc) was compared between 2 groups by two-sample t test. Results Compared with healthy controls, CBD group demonstrated asymmetrically decreased glucose metabolism mainly in the cere-bral hemisphere opposite to the more clinically affected body side, including the superior, middle and inferi-or frontal gyrus, the precentral gyrus, the superior and inferior parietal lobule, the angular gyrus, the supra-marginal gyrus, the precuneus, the middle occipital gyrus, the middle and inferior temporal gyrus, Heschl gyrus, the fusiform gyrus, the insula and the thalamus. And relatively increased glucose metabolism was present in ipsilateral precentral and postcentral gyrus, hippocampus, insula and putamen, bilateral cerebel-lum, paracentral lobules and pontine. The rCMRglc in those regions was significantly different between CBD patients and healthy controls (t values: 4.236-9.044, all P<0.01). Conclusion The asymmetric cerebral glucose metabolism features in CBD based on 18 F-FDG PET imaging contribute to the differential diagnosis between CBD patients and healthy subjects.

Dementia care is a chronic stressor severely influencing on physical and mental health and social life of family caregivers.In the research field of dementia care,the studies are inadequate regarding to the influence of caregiving stress on physical and mental health,especially on the risk of suffering from common chronic disease of family caregivers.So far,the studies are mainly based on hypotheses associated with chronic stress-induced three perspectives,i.e.,excessive activation of sympathetic nervous system,endothelial injury,and excessive activation of pro-inflammatory and procoagulant factors.This paper summarizes the research progress from these three perspectives.

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.