Objective:To investigate the protective effect of grape seed proanthocyanidin extract(GSPE) on fluoride-induced nephrotoxicity and study the possible role of the nuclear factor E2-related factor 2 (Nrf2) signaling pathway.Methods:Twenty-four male Sprague-Dawley (SD) rats (aged 8 weeks, weighing 240 - 260 g) were randomly divided into four groups using a random number table: control group, GSPE group, sodium fluoride (NaF) group, and NaF + GSPE group, with 6 rats in each group. The control group was fed a standard diet for 28 d. The GSPE group was administered GSPE (100 mg/kg) via gavage daily for 28 d. The NaF group was fed a standard diet for 7 d, followed by NaF solution (600 mg/kg) for 21 d. A drinking-water-borne fluorosis rat model was established. The NaF + GSPE group was pretreated with GSPE (100 mg/kg) for 7 d follwed by NaF solution (600 mg/kg) for 21 d. At the end of the experiment, the levels of renal oxidative stress were assessed by measuring malondialdehyde (MDA) and reduced glutathione (GSH) levels, catalase (CAT) and superoxide dismutase (SOD) activities. Renal function was evaluated by analyzing the levels of serum creatinine (Cr), blood urea nitrogen (BUN), and Urea. Renal pathological changes and apoptosis levels were examined using hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, respectively. Additionally, the expression levels of Nrf2 signaling pathway-related proteins [Nrf2, heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase-1 (NQO-1), and caspase-3] were detected by Western blot.Results:Compared with the control group, the NaF group had higher level of renal MDA, lower level of GSH, and lower activities of CAT and SOD( P < 0.001). Compared with the NaF group, the NaF + GSPE group had lower level of renal MDA, higher level of GSH, and higher activities of CAT and SOD ( P < 0.001). The serum Cr, BUN, and Urea levels were significantly elevated in the NaF group compared to the control group ( P < 0.001), while these parameters were significantly reduced in the NaF + GSPE group relative to the NaF group ( P < 0.001). The HE staining results showed that the glomerular volume increased and the basement membrane thickened; renal tubules disruption of cellular structure, with degeneration, dilation, or shedding of renal tubular epithelial cells; inflammatory cell infiltration can be seen in the renal interstitium in the NaF group of rats. Compared with the NaF group, the pathological changes in the NaF + GSPE group were significantly improved. The TUNEL results showed that the apoptosis rates of renal cells in the NaF and NaF + GSPE groups [(26.82 ± 2.25)%, (12.83 ± 1.32)%] were higher than those in the control group [(6.22 ± 0.62)%, P < 0.001], and in the NaF + GSPE group was lower than that in the NaF group ( P < 0.001). Compared with the control group, the NaF group showed expression levels of Nrf2 (nuclear), HO-1, NQO-1, activated caspase-3 protein were higher, while the expression level of Nrf2 (cytoplasmic) protein was lower. Compared with the NaF group, the NaF + GSPE group had higher expression levels of Nrf2 (nuclear, cytoplasmic), HO-1, and NQO-1 proteins, and lower expression level of activated caspase-3 protein ( P < 0.001). Conclusion:GSPE can alleviate the nephrotoxicity induced by fluoride, which may be related to activating the Nrf2 signaling pathway to enhance the body's antioxidant capacity and reduce the level of renal cell apoptosis.

Objective To investigate the application of cardiovascular magnetic resonance feature tracking(CMR-FT)in assessing myocardial strain in dilated cardiomyopathy(DCM)patients residing at high altitudes.Methods We retrospectively enrolled 29 DCM patients living at high altitudes(DCM-H),27 DCM patients living in a low-altitude plain environment(DCM-P),23 healthy volunteers living at a high altitude(HV-H),and 24 healthy volunteers living in a low-altitude plain environment(HV-P).All subjects underwent cine MRI scanning using a 3.0T rapid steady-state free precession sequence.The CMR images thus acquired were analyzed using cvi42,a post-processing software,to obtain left ventricular function and myocardial strain parameters.Results Compared with the HV-H group,the DCM-H group showed higher left ventricle end-diastolic volume(LVEDV)and left ventricle end-systolic volume(LVESV),and lower left ventricular ejection fraction(LVEF)and stroke volume(LVSV)(all P＜0.01).No significant difference was observed in cardiac function between the DCM-H and DCM-P groups(all P＞0.05).The absolute values of global radial strain(GRS),global circumferential strain(GCS),and global longitudinal strain(GLS)in the DCM-H group were lower than those in the HV-P group([14.5±6.5]％vs.[34.2±10.7]％,[-11.1±4.4]％vs.[-19.9±2.8]％,and[-7.7±3.2]％vs.[-13.6±4.1]％,respectively),with the differences being statistically significant(all P＜0.001).The DCM-H group had higher absolute GRS,GCS,and GCS values than the DCM-P group did([14.5±6.5]％vs.[7.0±2.7]％,[-11.1±4.4]％vs.[—5.4±2.2]％,and[—7.7±3.2]％vs.[—4.3±1.7]％,respectivley,all P＜0.01).Conclusion Myocardial strain in DCM patients living at a high altitude is lower than that in healthy volunteers living at a high altitude,but higher than that in DCM patients living in a low-altitude plain environment.CMR-FT can be used to quantitatively assess myocardial contractility in DCM patients living at a high altitude,showing promise for clinical application.

Purpose To quantitatively evaluate myocardial three-dimensional strains using cardiac magnetic resonance feature tracking,investigate whether ferroptosis contributes to myocardial injury in experimental autoimmune myocarditis(EAM),and explore the relationship between strains and ferroptosis biomarkers.Materials and Methods EAM was induced in Lewis rats via subcutaneous injections with porcine cardiac myosin emulsified in same amount of complete Freund's adjuvant.Both EAM(n=15)and control(n=15)rats underwent 7T cardiac magnetic resonance and were euthanatized at day 21.Cardiac magnetic resonance assessments included left ventricular function and strains.The cardiac damage and ferroptosis-related markers were measured with histopathology,serology,Western blot and polymerase chain reaction.Results The left ventricular ejection fraction declined in the rats post-immunization with porcine cardiac myosin at day 21(t=7.86,P<0.001).Global,basal,middle and apical strains encompassing radial strain,circumferential strain and longitudinal strain were significantly lower in the EAM group compared to the control group(t=2.61-10.45,P<0.05).Cardiac and inflammatory biomarkers,including lactate dehydrogenase,cardiac troponin I and interferon γ indicated myocardial injury in EAM group compared to the control group(t=16.54,21.20,16.06,all P<0.001).Prussian blue staining revealed iron overload in EAM group.Compared to the control group,malondialdehyde and reactive oxygen significantly increased in EAM group(t=15.75,3.47,both P<0.01),and superoxide dismutase were significantly decreased(t=14.21,P<0.001).Western blot and histopathology showed that the protein expression of ferritin heavy chain was significantly higher in the EAM group compared to the control group(t=5.15,P<0.01).The solute carrier family 7 member 11/glutathione/glutathione peroxidase 4 axis was down regulated at serum,mRNA and protein expression levels in EAM group(t=7.73-30.14,all P<0.001).Global circumferential and longitudinal strains were significantly associated with ferroptosis biomarkers(r=-0.854-0.829,all P<0.05).Conclusion Cardiac magnetic resonance feature tracking has the potential to quantitatively evaluate myocardial damage of EAM which involves ferroptosis.The strains are related with ferroptosis biomarkers.

Objective:To investigate the protective effect of grape seed proanthocyanidin extract(GSPE) on fluoride-induced nephrotoxicity and study the possible role of the nuclear factor E2-related factor 2 (Nrf2) signaling pathway.Methods:Twenty-four male Sprague-Dawley (SD) rats (aged 8 weeks, weighing 240 - 260 g) were randomly divided into four groups using a random number table: control group, GSPE group, sodium fluoride (NaF) group, and NaF + GSPE group, with 6 rats in each group. The control group was fed a standard diet for 28 d. The GSPE group was administered GSPE (100 mg/kg) via gavage daily for 28 d. The NaF group was fed a standard diet for 7 d, followed by NaF solution (600 mg/kg) for 21 d. A drinking-water-borne fluorosis rat model was established. The NaF + GSPE group was pretreated with GSPE (100 mg/kg) for 7 d follwed by NaF solution (600 mg/kg) for 21 d. At the end of the experiment, the levels of renal oxidative stress were assessed by measuring malondialdehyde (MDA) and reduced glutathione (GSH) levels, catalase (CAT) and superoxide dismutase (SOD) activities. Renal function was evaluated by analyzing the levels of serum creatinine (Cr), blood urea nitrogen (BUN), and Urea. Renal pathological changes and apoptosis levels were examined using hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, respectively. Additionally, the expression levels of Nrf2 signaling pathway-related proteins [Nrf2, heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase-1 (NQO-1), and caspase-3] were detected by Western blot.Results:Compared with the control group, the NaF group had higher level of renal MDA, lower level of GSH, and lower activities of CAT and SOD( P < 0.001). Compared with the NaF group, the NaF + GSPE group had lower level of renal MDA, higher level of GSH, and higher activities of CAT and SOD ( P < 0.001). The serum Cr, BUN, and Urea levels were significantly elevated in the NaF group compared to the control group ( P < 0.001), while these parameters were significantly reduced in the NaF + GSPE group relative to the NaF group ( P < 0.001). The HE staining results showed that the glomerular volume increased and the basement membrane thickened; renal tubules disruption of cellular structure, with degeneration, dilation, or shedding of renal tubular epithelial cells; inflammatory cell infiltration can be seen in the renal interstitium in the NaF group of rats. Compared with the NaF group, the pathological changes in the NaF + GSPE group were significantly improved. The TUNEL results showed that the apoptosis rates of renal cells in the NaF and NaF + GSPE groups [(26.82 ± 2.25)%, (12.83 ± 1.32)%] were higher than those in the control group [(6.22 ± 0.62)%, P < 0.001], and in the NaF + GSPE group was lower than that in the NaF group ( P < 0.001). Compared with the control group, the NaF group showed expression levels of Nrf2 (nuclear), HO-1, NQO-1, activated caspase-3 protein were higher, while the expression level of Nrf2 (cytoplasmic) protein was lower. Compared with the NaF group, the NaF + GSPE group had higher expression levels of Nrf2 (nuclear, cytoplasmic), HO-1, and NQO-1 proteins, and lower expression level of activated caspase-3 protein ( P < 0.001). Conclusion:GSPE can alleviate the nephrotoxicity induced by fluoride, which may be related to activating the Nrf2 signaling pathway to enhance the body's antioxidant capacity and reduce the level of renal cell apoptosis.

Purpose To quantitatively evaluate myocardial three-dimensional strains using cardiac magnetic resonance feature tracking,investigate whether ferroptosis contributes to myocardial injury in experimental autoimmune myocarditis(EAM),and explore the relationship between strains and ferroptosis biomarkers.Materials and Methods EAM was induced in Lewis rats via subcutaneous injections with porcine cardiac myosin emulsified in same amount of complete Freund's adjuvant.Both EAM(n=15)and control(n=15)rats underwent 7T cardiac magnetic resonance and were euthanatized at day 21.Cardiac magnetic resonance assessments included left ventricular function and strains.The cardiac damage and ferroptosis-related markers were measured with histopathology,serology,Western blot and polymerase chain reaction.Results The left ventricular ejection fraction declined in the rats post-immunization with porcine cardiac myosin at day 21(t=7.86,P<0.001).Global,basal,middle and apical strains encompassing radial strain,circumferential strain and longitudinal strain were significantly lower in the EAM group compared to the control group(t=2.61-10.45,P<0.05).Cardiac and inflammatory biomarkers,including lactate dehydrogenase,cardiac troponin I and interferon γ indicated myocardial injury in EAM group compared to the control group(t=16.54,21.20,16.06,all P<0.001).Prussian blue staining revealed iron overload in EAM group.Compared to the control group,malondialdehyde and reactive oxygen significantly increased in EAM group(t=15.75,3.47,both P<0.01),and superoxide dismutase were significantly decreased(t=14.21,P<0.001).Western blot and histopathology showed that the protein expression of ferritin heavy chain was significantly higher in the EAM group compared to the control group(t=5.15,P<0.01).The solute carrier family 7 member 11/glutathione/glutathione peroxidase 4 axis was down regulated at serum,mRNA and protein expression levels in EAM group(t=7.73-30.14,all P<0.001).Global circumferential and longitudinal strains were significantly associated with ferroptosis biomarkers(r=-0.854-0.829,all P<0.05).Conclusion Cardiac magnetic resonance feature tracking has the potential to quantitatively evaluate myocardial damage of EAM which involves ferroptosis.The strains are related with ferroptosis biomarkers.

Objective To establish a hypobaric hypoxia rat model in a real high-altitude environment,to investigate the effects of the real high-altitude environment on rat bone mass and bone microstructure using multiple methods such as Micro CT,blood biochemistry,and pathology,and to explore the potential mechanisms involved.Methods Sprague Dawley(SD)rats were transported to the Yushu Plateau Laboratory(at 4250 m above sea level)in Qinghai Province and kept there for 4,or 8,or 18 months.These groups were designated as H-4,H-8,and H-18,respectively.Upon completion of the high-altitude exposure,these animals were transported to the Molecular Imaging Laboratory,West China Hospital,Sichuan University(at 500 m above sea level)in Chengdu for relevant testing and comparison with the control animals raised in a low-altitude environment for the same durations(designated L-4,L-8,and L-18).The tests performed included blood biochemistry,Micro CT imaging,and pathological assessments such as ELISA,Western blot,and HE and TRAP staining.Results Compared with that of the control group,the body mass of rats in the H-4 and H-18 groups decreased significantly(H-4 group vs.L-4 group:[513.75±35.10]g vs.[649.18±60.03]g,P＜0.01;H-18 group vs.L-18 group:[535.58±66.65]g vs.[670.86±44.96]g,P＜0.01).The serum Ca2+concentration was higher in the H-8 group and H-18 group compared to that in the control group(H-8 group vs.L-8 group:[2.48±0.09]mmol/L vs.[2.38±0.07]mmol/L,P＜0.05;H-18 group vs.L-18 group:[2.55±0.11]mmol/L vs.[2.13±0.27]mmol/L,P＜0.05).No statistically significant difference was observed in the concentration of P3+.Bone metabolism indicator cross-linked carboxy-terminal telopeptide of type Ⅰ collagen(CTX-Ⅰ)was significantly increased in all high-altitude groups compared to the low-altitude groups(H-4 group vs.L-4 group:[1.44±0.08]ng/mL vs.[0.70±0.13]ng/mL,P＜0.01;H-8 group vs.L-8 group:[1.52±0.10]ng/mL vs.[0.75±0.10]ng/mL,P＜0.01;H-18 group vs.L-18 group:[2.70±0.13]ng/mL vs.[1.94±0.15]ng/mL,P＜0.01).In addition,CT results showed a decrease in bone volume fraction of trabecular bone in the three high-altitude groups(H-4 group vs.L-4 group:[7.48±2.35]％vs.[10.40±2.93]％,P＜0.05;H-8 group vs.L-8 group:[7.17±2.68]％vs.[10.09±2.95]％,P＜0.05;H-18 group vs.L-18 group:[2.90±2.91]％vs.[8.68±4.11]％,P＜0.01),and increased trabecular separation in the three high-altitude groups(H-4 group vs.L-4 group:[0.70±0.12]mm vs.[0.60±0.06]mm,P＜0.05;H-8 group vs.L-8 group:[0.68±0.07]mm vs.[0.59±0.05]mm,P＜0.01;H-18 group vs.L-18 group:[0.80±0.09]mm vs.[0.70±0.09]mm,P＜0.05).TRAP staining showed an increase in osteoclasts in the H-4 and H-18 groups.Western blot results indicated an increase in the expression of receptor activator of nuclear factor-κB ligand(RANKL)and hypoxia inducible factor-1α(HIF-1α)in high-altitude environment,while the expression of osteoprotegerin(OPG)was inhibited.Conclusion The impact of high-altitude environment on rat femurs is characterized primarily by a reduction in trabecular bone mass and damage to bone microstructure.

Objecfive Most prostate cancer contains two or more widely separate turnors.To study the origin of prostate cancer based on the analysis of microsatellite alteration in separate tumors from the same prostate.Methods A polymerase chain reaction (PCR) was used to examine the allelic loss pattern of 4 microsatellite polymorphic markers on chromosome 8p (D8S133,D8S136,D8S137) and 17q (D17S855) in multifocal tumors of prostate from 19 patients.DNA samples were obtained from different regions of distinctly separate tumors on single case using microdissection technique.Results The overall frequence of LOH at D8S133,D8S136,D8S137 and D17S855 for all informative cases was 74%,38%,86% and 46%respectively in 40 separate tumors of prostate from 19 patients.The pattern of allelic loss was not identical in 15 of 18 (83%) informative cases. It showed that the different regions of prostate cancer were independent origin respectively.Discordant pattern of histology was observe in distantly separate regions.whereas the same allele was consistently lost in samples from different regions of the same tumor in 3 cases. Condusion Current data supports independent origin of multiple tumors in most prostate cancer patients.

Objective To investigate expression of CD56, CD95(Fas), Ki-67, p53, bcl-2, HMB45 and S-100 in mucosa amelanotic malignant melanoma in order to improve the pathological diagnosis level reduce wrong diagnosis and avoid missing dignosis, and afford objective factors for prognosis and therapy. Methods The techniques of tissues chips and immunohistochemical lablling were used for analyzing 48 cases of mucosa amelanotic malignant melanoma. Results The positive rates of HMB45 and S-100 were 100 % (48/48) and 85.4 % (41/48) respectively. The positive rate of CD56 was 91.6 % (44/48), there was not statistical difference between original cases and metastatic cases. The positive rate of CD95 was 85.4 %(41/48). In which it is 100 % (11) in 11 cases of having lymphanoid metastasis. The positive rates of Ki-67 and p53 were 79.2 % (38/48) and 58.3 % (28/48) respectively. The positive distribution of Ki-67 was almost same as CD95. The positive rate of bcl-2 was 39.6 % (19/48), the positive expression was significantly different between p53 and bcl-2(P