1.Experimental Study of Tongluo Yizhi Formula Attenuating Memory Deficits in Rat Model of Vascular Dementia through PERK-mediated Endoplasmic Reticulum Stress Pathway
Dali XUE ; Xiaozhe ZHANG ; Hui WANG ; Guanfeng MA ; Supeng GUO
Journal of Modern Laboratory Medicine 2025;40(1):17-23
Objective To observe the effect of self-formulated Tongluo Yizhi formula on memory deficits and endoplasmic reticulum stress (ERS) in rats with vascular dementia (VD) and its mechanism. Methods Fifty SPF-grade Wistar rats were randomly divided into sham surgery group,model group,TLYZF low-dose (12.5 mg/kg/d) group,TLYZF medium dose (25 mg/kg/d) group and TLYZF high-dose (50 mg/kg/d) group,with 10 rats in each group. Except for the Sham group,other rats were subjected to modified bilateral carotid artery ligation to construct VD models. After 4 weeks of TLYZF intervention,Morris water maze experiment was used to evaluate the learning and memory abilities of rats.Hematoxylin eosin staining was used to detect the pathological structure of the hippocampus.Nissl staining was used to detect hippocampal neuronal damage.In situ end transfer enzyme labeling staining was used to detect hippocampal neuronal apoptosis.Enzyme-linked immunosorbent assay was used to detect the content of inflammatory factors in hippocampal tissue.Protein immunoblotting was used to detect apoptosis of hippocampal tissue cells and the expression of ERS related proteins.Results Compared with the Sham group,the average escape latency of the model group was prolonged(t=14.059),the number of crossing the platform was reduced(t=8.534),the hippocampal neurons were disorderly arranged,the morphology was fuzzy,the cytoplasmic vacuolization,the nuclear pyknosis,the inflammatory cell infiltration increased,the number of Nissl bodies decreased(t=17.131),and the neuronal apoptosis rate increased(t=17.701). The contents of tumor necrosis factor-α (TNF-α),interleukin-1 β (IL-1β) and IL-6 increased(t=6.541,6.957,10.014),the expression of Bcl-2 associated X protein (Bax),cleaved caspase-3,glucose-regulated protein 78 (GRP78),phosphorylated endoplasmic reticulum kinase (p-PERK)/PERK,phosphorylated eukaryotic initiation factor 2α (p-eIF2α)/eIF2α,activating transcription factor 4 (ATF4) and CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) increased(t=13.548~76.468),while the expression of B-cell lymphoma-2 (Bcl-2) proteinwas decreased (t=39.691),and the difference were statistically significant (all P<0.05),respectively. Compared with the model group,the average escape latency of TLYZF-treated rats was shortened(t=2.476,7.266,11.306),the number of platform crossing was increased(t=2.187,4.471,6.932),the hippocampal neurons were relatively regular,the nucleus was clear,the inflammatory cell infiltration was significantly reduced,the content of Nissl bodies was increased(t=4.359,9.477,11.449),the apoptosis rate of neurons was decreased(t=3.631,6.145,7.580),the contents of TNF-α,IL-1β and IL-6 were decreased (t=2.382~8.293),the expression of Bax(t=4.696,16.250,20.250),cleaved caspase-3(t=15.205,27.000,26.833),GRP78(t=5.918,13.139,13.741),p-PERK/PERK(t=13.416,14.230,17.889),p-eIF2α/eIF2α(t=20.152,39.346,50.750),ATF4(t=12.093,24.395,22.946)and CHOP protein((t=21.592,19.207,22.136)decreased,while Bcl-2 protein increased(t=7.474,20.761,35.350),the differences were statistically significant (all P<0.05),respectively,and the high-dose group had the best therapeutic effect.Conclusion Self-formulated Tongluo Yizhi formula could reduce the apoptosis and inflammatory injury of hippocampal neurons in VD rats,and its mechanism may be related to inhibiting ERS mediated by PERK.
2.Experimental Study of Tongluo Yizhi Formula Attenuating Memory Deficits in Rat Model of Vascular Dementia through PERK-mediated Endoplasmic Reticulum Stress Pathway
Dali XUE ; Xiaozhe ZHANG ; Hui WANG ; Guanfeng MA ; Supeng GUO
Journal of Modern Laboratory Medicine 2025;40(1):17-23
Objective To observe the effect of self-formulated Tongluo Yizhi formula on memory deficits and endoplasmic reticulum stress (ERS) in rats with vascular dementia (VD) and its mechanism. Methods Fifty SPF-grade Wistar rats were randomly divided into sham surgery group,model group,TLYZF low-dose (12.5 mg/kg/d) group,TLYZF medium dose (25 mg/kg/d) group and TLYZF high-dose (50 mg/kg/d) group,with 10 rats in each group. Except for the Sham group,other rats were subjected to modified bilateral carotid artery ligation to construct VD models. After 4 weeks of TLYZF intervention,Morris water maze experiment was used to evaluate the learning and memory abilities of rats.Hematoxylin eosin staining was used to detect the pathological structure of the hippocampus.Nissl staining was used to detect hippocampal neuronal damage.In situ end transfer enzyme labeling staining was used to detect hippocampal neuronal apoptosis.Enzyme-linked immunosorbent assay was used to detect the content of inflammatory factors in hippocampal tissue.Protein immunoblotting was used to detect apoptosis of hippocampal tissue cells and the expression of ERS related proteins.Results Compared with the Sham group,the average escape latency of the model group was prolonged(t=14.059),the number of crossing the platform was reduced(t=8.534),the hippocampal neurons were disorderly arranged,the morphology was fuzzy,the cytoplasmic vacuolization,the nuclear pyknosis,the inflammatory cell infiltration increased,the number of Nissl bodies decreased(t=17.131),and the neuronal apoptosis rate increased(t=17.701). The contents of tumor necrosis factor-α (TNF-α),interleukin-1 β (IL-1β) and IL-6 increased(t=6.541,6.957,10.014),the expression of Bcl-2 associated X protein (Bax),cleaved caspase-3,glucose-regulated protein 78 (GRP78),phosphorylated endoplasmic reticulum kinase (p-PERK)/PERK,phosphorylated eukaryotic initiation factor 2α (p-eIF2α)/eIF2α,activating transcription factor 4 (ATF4) and CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) increased(t=13.548~76.468),while the expression of B-cell lymphoma-2 (Bcl-2) proteinwas decreased (t=39.691),and the difference were statistically significant (all P<0.05),respectively. Compared with the model group,the average escape latency of TLYZF-treated rats was shortened(t=2.476,7.266,11.306),the number of platform crossing was increased(t=2.187,4.471,6.932),the hippocampal neurons were relatively regular,the nucleus was clear,the inflammatory cell infiltration was significantly reduced,the content of Nissl bodies was increased(t=4.359,9.477,11.449),the apoptosis rate of neurons was decreased(t=3.631,6.145,7.580),the contents of TNF-α,IL-1β and IL-6 were decreased (t=2.382~8.293),the expression of Bax(t=4.696,16.250,20.250),cleaved caspase-3(t=15.205,27.000,26.833),GRP78(t=5.918,13.139,13.741),p-PERK/PERK(t=13.416,14.230,17.889),p-eIF2α/eIF2α(t=20.152,39.346,50.750),ATF4(t=12.093,24.395,22.946)and CHOP protein((t=21.592,19.207,22.136)decreased,while Bcl-2 protein increased(t=7.474,20.761,35.350),the differences were statistically significant (all P<0.05),respectively,and the high-dose group had the best therapeutic effect.Conclusion Self-formulated Tongluo Yizhi formula could reduce the apoptosis and inflammatory injury of hippocampal neurons in VD rats,and its mechanism may be related to inhibiting ERS mediated by PERK.
3.Literature Analysis of the Preparation Elements of Animal Models of Skin Photoaging and the Data of Subjects
Yasheng DENG ; Jiang LIN ; Chiling GAN ; Guanfeng ZENG ; Jiayin HUANG ; Huifang DENG ; Yingxian MA ; Siyin HAN
Laboratory Animal and Comparative Medicine 2023;43(4):406-414
Objective To analyze the modeling elements and subjects of the animal model of skin photoaging, and to provide a reference for the preparation and improvement of the model and a basis for the scientific evaluation of the subject.Methods By searching and collecting relevant literature on the preparation of animal models of skin photoaging from 2010 to 2022 in the China National Knowledge Infrastructure, Wanfang Database, and PubMed database, the model animal species, gender, modeling method, modeling cycle, radiation source and its distance from the modeling site, cumulative radiation volume, detection indicators, and subjects (drugs or treatments) recorded in the literature were collated and summarized, and a database was established for statistical analysis.Results 257 articles that met the inclusion criteria were selected. Among them, the most common animal model was SKH-1 hairless mice, followed by SD rats and KM mice; the gender of animals was mainly female, medium-wave ultraviolet B (UVB) was often used as the radiation source, the distance between the radiation source and the modelling site was mostly 30 cm, and the modelling period was usually 40-60 days. The cumulative dose of long-wave ultraviolet A (UVA) was between 100-150 J/cm2, and the cumulative dose of UVB was between 5-10 J/cm2. The tests used after model establishment were skin histopathological examination, skin tissue homogenization, fibre staining, immunoblotting, etc. Subjects included Chinese herbal medicines, Chinese herbal extracts, Chinese patent medicines, Chinese herbal compound medicines, chemical drugs, biological agents and other treatments, while the animal model of skin photoaging was also used for clinical efficacy studies of external Chinese medicine, physiotherapy and positive control drugs.Conclusion In skin photoaging animal experiments, female SKH-1 hairless mice are often used, and UVB is used as the radiation source. The modeling period is usually 40-60 days, and the minimum erythema dose (MED) is incremented week by week. The cumulative UVB irradiation dose ranges from 0 to 10 J/cm2, which has the advantages of high success rate, good reproducibility and high similarity with clinical disease.
4.Clinical efficacy analysis of flexible ureteroscopic lithotripsy combined with percutaneous nephrolithotomy treating for patients with staghorn calculi
Guanfeng WANG ; Ru CHEN ; Feng MA
Journal of Clinical Surgery 2017;25(4):304-306
Objective To investigate the clinical efficacy of flexible ureteroscopic lithotripsy combined with percutaneous nephrolithotomy treating for partial staghorn calculi.Methods 84 patients diagnosed as partial staghorn calculi in our hospital were randomly divided into group A and B with each group 42 patients.Patients in group A received the conventional minimally invasive percutaneous nephrolithotomy in the prone position,and patients in group B received the percutaneous nephrolithotomy combined with flexible ureteroscopic lithotripsy in the modified Valdivia position.The post-operative stone free rate and complications were recorded.Results The surgery time in group B was longer than that in group A [(106.44±18.46)min vs(83.69±10.29)min],with statistically significant difference(P<0.05).However,the first stone free rate in group B was higher than that in group A(85.71% vs 59.52%,P<0.05),and the blood loss in group B was lower than that in group A [(70.02±9.15)ml vs(87.41±9.89)ml,P<0.05].The common complication of patients in two groups was fever(temperature >38.5℃),but there was no notably difference between the two group(P>0.05).Conclusion Compared with the regular percutaneous nephrolithotomy,flexible ureteroscopic lithotripsy combined with percutaneous nephrolithotomy treating partial staghorn calculi has the shorter operation time,the less blood volume and the higher first stone free rate.Furthermore,the combination method did not significantly increasing the incidence of patient's complication.

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