Aim To investigate the protective effects of echinacoside ( ECH ) on oxidative stress injury in vascular dementia rats. Methods Vascular dementia model was duplicated by means of permanent ligation of bilateral common carotid artery at two times intervals for three days. Biochemical methods was used to detect the GSH, NO, GSH-Px, NOS in each group rat’ s cor-tex and hippocampus. The change of the tissue struc-ture in CA1 area of hippocampus in each group was ob-served and analyzed by microscope after HE staining. Results Compared with sham group, the content of GSH and activity of GSH-Px in the rats of the model group were decreased significantly ( P<0. 01 ) , while the content of NO and activity of NOS were increased obviously (P<0. 05, P <0. 01). Meanwhile, in tis-sue structure of rat’ s hippocampal CA1 area, the num-bers of cells were reduced, cell arrangement was sparse and in disorder, cell form was not intact and cell struc-ture was not normal, cytoplasm, nucleus and cyto-plasm were blurry, nuclei were hyperchromatic and in contraction, and were changed to triangular or irregular shape, lacking nucleoli, and proliferation of glial cells appeared. Compared with the model group, the content of GSH and activity of GSH-Px in the rats of the treat-ment groups were increased significantly ( P<0. 05 , P<0 . 01 ) , while the activity of NOS was decreased ob-viously (P<0. 05, P<0. 01). Meanwhile, in rat tis-sue structure of hippocampal CA1 area, the rat cells in each treatment group were arranged in order, the form of structure was normal, the nuclei were less hyper-chromatic and contracted. Compared with Gal group and sham group, the rats of ECH high dosage group were not different in every part of experiment ( P >0. 05 ) . Conclusion ECH exerts protection on oxida-tive stress injury in vascular dementia rats.

Aim To investigate the effects of echina-coside ( ECH ) on the learning and memory capacities and brain level of oxygen free radicals of rats with de-mentia induced by amyloid β-peptide. Methods Six-ty Sprague Dawley rats, weighing (300±10) g, were randomly divided with 10 rats pergroup into 6 groups:sham operated group, model, ECH high dose (40 mg ·kg-1·d-1), ECH middle dose (20 mg· kg-1· d-1), ECH low dose (10 mg·kg-1·d-1) and Hup A (Huperzine A, 0. 02 mg·kg-1·d-1) group. Mor-ris maze tests were conducted for evaluating the learn-ing and memory ability. Content of malondialdehyde (MDA), nitric oxide (NO) and activities of superox-ide dismutase ( SOD) and NO synthase ( NOS) in the hippocampus and cortex were detected. ResultsAβ25-35 induced significant learning and memory im-pairment in the rats. Compared with the rats in model group, those treated with ECH at different doses all manifested alleviation of learning and memory impair-ment ( P<0 . 01 , P<0 . 05 ) . Cotents of MDA of ECH treatment group were obviously decreased, while SOD activities were obviously increased, and significantly reduced the release of NO and NOS in the hippocam-pus and cortex brain tissue ( P <0 . 01 , P <0.05 ) . Conclusion ECH can enhance the learning and mem-ory ability in rats with AD, which is presumably relat-ed to accelerating the cleaning of oxygen free radicals and reducing oxidative stress in brain of AD rats.

Aim Tostudytheeffectsofechinacoside on striatal and hippocampus extracellular fluid of mono-amine neurotransmitter in Parkinsonˊs disease rat, and discuss the possible mechanism of the ECH effect on brainprotection.Methods Doublepointinjectionof 6-hydroxy dopamine damaged was adopted to make PD model, the corresponding drugs or normal saline were intraperitoneally injected for 4 consecutive weeks, mi-cro-dialysis program in double targets was taken after treatment, the dialysate was injected into HPLC-EDC, and striatal and hippocampus extracellular fluid of DA, DOPAC, HVA, NA, 5-HT levels of each group were measured. Results Compared with control group, striatal and hippocampus extracellular fluid of DA, DOPAC, HVA, NE, 5-HT levels were significantly re-duced in model group ( P <0. 01 ); compared with model group, striatal and hippocampus extracellular fluid of DA, DOPAC, HVA, NE, 5-HT levels were significantly increased in treatment group ( P <0 . 05 , P<0. 01 ) , and the content of five kinds of substances in ECH high dose group was similar to that in MD group.Conclusion ECHcouldincreaseextracellular monoamine neurotransmitters in striatum and hippo-campus, ECH has a therapeutic effect on PD, and echinacoside is possible mechanisms of the brain pro-tective effect of PD rats.

Objective To explore the neuroprotective role of Genistein (GEN) on hippocampal CA1 neurons and the possible mechanism following global cerebral ischemia ( GCI) in rats.Methods Seventy five rats were subjected to global cerebral ischemia ( GCI ) by four-vessel occlusion and randomly divided into five groups , sham, ischemia/reperfusion (I/R), GEN, ICI 182,780 and vehicle groups.Fluoro-Jade B and neuron-specific nuclear-binding protein ( NeuN) staining was used to observe CA 1 neuronal survival .TUNEL was used to detect apoptotic neurons .Spatial learning and memory function of the rats were evaluated by Morris water maze .Results The best dose of neuroprotective role of GEN was 1.0mg/kg body weight.Compared with sham, TUNEL-positive neurons in the hippocampal CA1 region increased significantly in I/R and vehicle groups (P<0.01), while post-treatment with GEN (1.0mg/kg) at 5min after ischemia by tail vein injection decreased markedly (P<0.01).Treatment of 1.0mg/kg GEN markedly attenuated spatial learning and memory deficits of the rats after ischemic insult compared to I /R group.Furthermore, ICI 182,780 significantly abolished the neuroprotective role of GEN (P <0.01).Conclusion The low-dose (1.0mg/kg) GEN significantly attenuates neuronal damage and cognitive deficits following GCI in rats , and the mechanism may be involved in estrogen receptor activity.

Aim To investigate the influence of echi-nacoside ( ECH ) on monoaminergic neurotransmitter extracellular of the hippocampus and cerebral cortex in rat model of Alzheimer′s disease ( AD) , and ultimately to provide a theoretical basis for ECH′s improving the ability of learning and memory. Methods 60 male SD rats were randomly divided into 6 groups with 10 rats in each group : sham operation group, model, ECH groups of low, medium and high doses (10, 20, 40μg ·g-1 ·d-1 ) , and Hup A ( Huperzine A, 0. 02 μg· g-1·d-1) group. The AD rat model was established by abdominal cavity injection with D-galactose and uni-laterally injected with amyloid beta-protein fragment 25-35 ( Aβ25-35 ) into the right hippocampus. Morris wa-ter maze test was used to study the animals′ ability of spatial learning and memory. The synchronous dual-probe dual-channel brain microdialysis sampling tech-nology was applied to collect dialysates from different encephalic areas continuously, and combined with HPLC electrochemical detection were used to measure the extracellular levels of norepinephrine ( NE) , dopa-mine (DA), 5-serotonin (5-HT). Results 1. Com-pared with the sham operation group, the mean escape latency of the model group was significantly prolonged ( P<0. 05 ) , and the time that rats were in the plat-form quadrant was significantly shortened ( P<0. 05 );on the contrary, compared with the model group, the mean escape latency of ECH groups were significantly shortened ( P<0. 05 ) , and the time that rats were in the platform quadrant was significantly extended ( P<0. 05). 2. Compared with the sham operation group, the contents of NE,DA and 5-HT were significantly de-creased in the model group ( P <0. 05 ) . However, compared with the model group , ECH could improve the concentrations of NE, DA, 5-HT in the hippocam-pus and cerebral cortex, and these monoamine levels of the brain regions were restored to near control. Con-clusion ECH can effectively improve the ability of learning and memory of rats with AD, giving a rise to the monoamine neurotransmitter both in hippocampus and cortex, exerting a positive effect on treatment of cognitive dysfunction . The ECH low dose group is sig-nificantly lower than the ECH groups of medium and high doses and Hup A group in improving the ability of learning and memory.

Objective:To design Keap1-tat peptide and explore its neuroprotective role on hipocampal CA1 neuron,as well as the effect on spacial learning and memory function following global cerebral ische-mia.Methods:Adult male Sprague Dawley (SD)rats were subjected to global cerebral ischemia (GCI) by four-vessel occlusion for 1 5 min and randomly divided into five groups:sham,sham+Keap1-tat,is-chemia/reperfusion (I/R),Keap1-tat peptide-and vehicle-administrated groups.For Keap1-tat or vehi-cle groups,the rats were treated with Keap1-tat (30,50,1 00 μg in 5 μL 0.9%saline)or the same vo-lume vehicle by intracerebroventricular injection (icv)30 min prior to ischemia.Cresyl violet staining was used to observe the surviving neurons and 4-hydroxy-2-noneal (4-HNE ) and 8-hydroxy-2′-deox-yguanosine (8-OHdG)immunostaining were used to detect the change of markers response to oxidative stress in hippocampal CA1 region.The spatial learning and memory function of the rats was evaluated using Morris water maze.Results:Compared with sham group,the number of surviving neurons in ische-mia-reperfusion and vehicle groups significantly decreased in the hippocampal CA1 region (P<0.05 ), while administration of Keap1-tat significantly decreased the damage following GCI (P<0.05),and the dose of 50 μg existed the most effective neuroprotective role.Furthermore,immunostaining intensity of 4-HNE and 8-OHdG,markers of oxidative stress damage attenuated by Keap1-tat peptide as compared with vehicle group in CA1 region.Of significant interest,the time of finding underwater platform in Keap1-tat group animals was significantly short,and after removing the platform,the probe time of Keap1-tat group animals in the original quadrant where the platform was significantly increased compared with that of vehi-cle and I/R group animals (P<0.05).Conclusion:Keap1-tat peptide can effectively attenuate neuro-nal damage in hippocampal CA1 region and improve learning and memory function,which might bedue to the attenuation of oxidative stress caused by GCI.

The aim of this study is to investigate the effect of echinacoside (ECH) on cholinergic neurotransmitter extracellular of hippocampus and striatum and its possible mechanisms of neuro-protective effect against vascular dementia rats. In this study brain microdialysis technique combined with HPLC-IMER-ECD (high-performance liquid chromatography-immobilized enzyme reactor-electrochemical detector) was used. The bilateral common carotid arteries occluded in two times operation at 72 h interval for vascular dementia model rats were used and the successful vascular dementia model rats were examined by Morris water maze. The content of acetylcholine (ACh) and choline (Ch) of microdialysate extracellular of hippocampus and striatum was determined by HPLC-IMER-ECD and the AChE activity in the hippocampus was measured. The results showed that the success rate of vascular dementia model was 83.08% after six weeks; the results also showed that echinacoside and galantamine could increase the content of ACh and reduce the content of Ch extracellular of hippocampus and striatum significantly and the AChE activity increased significantly compared with that of the model group. The results suggested that echinacoside could promote the recovery of cholinergic neurotransmitter levels in vascular dementia rats' brain, which may be one of the mechanisms of neuro-protection.

Objective:To investigate the effect of laboratory indicators on hair loss in patients with female pattern hair loss (FPHL).Methods:Patients with FPHL who visited the Outpatient Clinic of the Department of Medical Aesthetics in Hangzhou First People’s Hospital from November 2022 to November 2023 were selected as the study group, and healthy women who matched the age of the study group in the physical examination center during the same period were selected as the control group. The general information of the patient was recorded, and was also tested by trichoscopy to rule out other patterns of alopecia. Representative indicators including testosterone, dehydroepiandrosterone sulfate(DHEA-S), thyroid-stimulating hormone, 25-hydroxyvitamin D, and serum ferritin were selected from laboratory tests for further analysis. Otherwise, the proportion of deficiency in vitamin D(<20 ng/ml) was calculated based on 25-hydroxyvitamin D levels (number of deficiency cases/total number of cases in each group×100%). Count data were presented as samples (percentages), and chi-square test was used for comparison between groups. Normally distributed continuous data were presented with Mean±SD, independent samples t-test was used for comparison between groups, M( Q1, Q3) was used for non-normally distributed continuous data, and Wilcoxon rank-sum test was used for comparison between groups. Multivariate logistic regression was used to analyze the influencing factors of FPHL. P<0.05 was statistically significant. Results:A total of 37 patients were selected in both groups. The mean age was (28.8±1.3) years in the study group and (29.6±0.9) years in the control group ( t=0.49, P=0.625). The body mass index was (22.8±0.4) kg/m 2 in the study group, and (23.5±0.3) kg/m 2 in the control group ( t=1.26, P=0.211). The testosterone level was 0.58 (0.49, 0.79) nmol/L in the study group, and 0.54 (0.50, 0.78) nmol/L in the control group( Z=1.42, P=0.157). The level of DHEA-S was 6.21 (5.18, 9.60) μmol/L in the study group, and 6.20 (5.20, 9.34) μmol/L in the control group ( Z=2.75, P=0.006). The level of thyroid-stimulating hormone was 2.56 (1.55, 3.66) mU/L in the study group and 1.49 (1.05, 2.65) mU/L in the control group ( Z=2.51, P=0.012). The level of 25-hydroxyvitamin D was 15.44 (11.80, 21.20) ng/ml in the study group, and the level of 25-hydroxyvitamin D was 20.32 (12.07, 21.20) ng/ml in the control group ( Z=2.30, P=0.021), and the proportion of 25-hydroxyvitamin D deficiency in the study group was 64.9% (24/37), which was higher than that in the control group [40.5% (15/37)] ( χ2=4.39, P=0.036). The serum ferritin level was 64.44 (39.47, 133.45) μg/L in the study group and 67.75 (52.63, 143.83) μg/L in the control group ( Z=0.70, P=0.484). The results of multivariate logistic regression analysis showed that the risk of FPHL was increased by the high level of DHEA-S and thyroid-stimulating hormone, and the low level of 25-hydroxyvitamin D (all P<0.05). Conclusion:Abnormal level of DHEA-S, thyroid-stimulating hormone, and 25-hydroxyvitamin D may be risk factors for FPHL.

Objective To investigate the improvement effect and related mechanism of roxadustat on myocardial ischemia-reperfusion(I/R)injury in mice.Methods Twenty four male C57BL/6N mice were randomly divided into the sham operation group,the control group and the roxadustat group,with eight mice in each group.A mouse myocardial I/R model was established.The control group was given 100 μL saline injection containing 5%dimethyl sulfoxide by gavage.The roxadustat group was given 25 mg/kg roxadustat by gavage.The left anterior descending coronary artery of mice in both groups was ligated for 40 minutes,and then reperfusion for 24 hours to establish the myocardial I/R model.In the sham operation group,only the left anterior coronary artery was pierced without ligation.The area of myocardial infarction in mice was detected by triphenyltetrazolium chloride(TTC)staining.The apoptosis of mouse cardiomyocytes was detected by TdT-mediated dUTP nick and labeling(TUNEL)staining.The expression of apoptosis-related proteins bcl-2 associated X protein(Bax),Caspase3 and inflammatory cell markers F4/80 and myeloperoxidase(MPO)were detected by immunohistochemistry staining.The damage of myocardial cells was observed by hematoxylin-eosin(HE)staining.Results The area of myocardial infarction after myocardial I/R was reduced in the roxadustat group compared to the control group and the sham operation group(P＜0.05).The number of apoptotic cells was higher in the control group and the roxadustat group than that in the sham operation group,and the number of apoptotic cells was lower in the roxadustat group than that in the control group(P＜0.05).The expression levels of Bax and Caspase3 proteins in myocardial tissue were higher in the control group and the roxadustat group than those in the sham operation group,while those of the roxadustat group was lower than those of the control group(P＜0.05).The expression levels of F4/80 and MPO proteins in myocardial tissue were lower in the roxadustat group than those in the control group(P＜0.05).In the control group,the myocardial tissue arrangement was disordered,and there was an increase in interstitial vacuoles.Compared with the control group,the myocardial cells were arranged more neatly in the roxadustat group,and the interstitial vacuoles were reduced.Conclusion Roxadustat can reduce the myocardial infarction area after I/R injury,inhibit myocardial cell apoptosis,alleviate myocardial injury,reduce infiltration of myocardial macrophages and neutrophils,and reduce inflammatory injury.

Objective:To investigate the effect of laboratory indicators on hair loss in patients with female pattern hair loss (FPHL).Methods:Patients with FPHL who visited the Outpatient Clinic of the Department of Medical Aesthetics in Hangzhou First People’s Hospital from November 2022 to November 2023 were selected as the study group, and healthy women who matched the age of the study group in the physical examination center during the same period were selected as the control group. The general information of the patient was recorded, and was also tested by trichoscopy to rule out other patterns of alopecia. Representative indicators including testosterone, dehydroepiandrosterone sulfate(DHEA-S), thyroid-stimulating hormone, 25-hydroxyvitamin D, and serum ferritin were selected from laboratory tests for further analysis. Otherwise, the proportion of deficiency in vitamin D(<20 ng/ml) was calculated based on 25-hydroxyvitamin D levels (number of deficiency cases/total number of cases in each group×100%). Count data were presented as samples (percentages), and chi-square test was used for comparison between groups. Normally distributed continuous data were presented with Mean±SD, independent samples t-test was used for comparison between groups, M( Q1, Q3) was used for non-normally distributed continuous data, and Wilcoxon rank-sum test was used for comparison between groups. Multivariate logistic regression was used to analyze the influencing factors of FPHL. P<0.05 was statistically significant. Results:A total of 37 patients were selected in both groups. The mean age was (28.8±1.3) years in the study group and (29.6±0.9) years in the control group ( t=0.49, P=0.625). The body mass index was (22.8±0.4) kg/m 2 in the study group, and (23.5±0.3) kg/m 2 in the control group ( t=1.26, P=0.211). The testosterone level was 0.58 (0.49, 0.79) nmol/L in the study group, and 0.54 (0.50, 0.78) nmol/L in the control group( Z=1.42, P=0.157). The level of DHEA-S was 6.21 (5.18, 9.60) μmol/L in the study group, and 6.20 (5.20, 9.34) μmol/L in the control group ( Z=2.75, P=0.006). The level of thyroid-stimulating hormone was 2.56 (1.55, 3.66) mU/L in the study group and 1.49 (1.05, 2.65) mU/L in the control group ( Z=2.51, P=0.012). The level of 25-hydroxyvitamin D was 15.44 (11.80, 21.20) ng/ml in the study group, and the level of 25-hydroxyvitamin D was 20.32 (12.07, 21.20) ng/ml in the control group ( Z=2.30, P=0.021), and the proportion of 25-hydroxyvitamin D deficiency in the study group was 64.9% (24/37), which was higher than that in the control group [40.5% (15/37)] ( χ2=4.39, P=0.036). The serum ferritin level was 64.44 (39.47, 133.45) μg/L in the study group and 67.75 (52.63, 143.83) μg/L in the control group ( Z=0.70, P=0.484). The results of multivariate logistic regression analysis showed that the risk of FPHL was increased by the high level of DHEA-S and thyroid-stimulating hormone, and the low level of 25-hydroxyvitamin D (all P<0.05). Conclusion:Abnormal level of DHEA-S, thyroid-stimulating hormone, and 25-hydroxyvitamin D may be risk factors for FPHL.