1.6-Shogaol alleviates cerebral injury after cardiac arrest-cardiopulmonary resuscitation in rats by inhibiting death-associated protein kinase 1-mediated autophagy.
Ouyang RAO ; Shixin LI ; Ning ZHU ; Hangxiang ZHOU ; Jie HU ; Yun LI ; Junling TAO ; Yehong LI ; Ying LIU
Chinese Critical Care Medicine 2025;37(6):568-575
OBJECTIVE:
To observe the neuroprotective effect of 6-shogaol (6-SH) in global cerebral ischemia/reperfusion injury (CIRI) following cardiac arrest (CA) and cardiopulmonary resuscitation (CPR) in rats.
METHODS:
Computer-aided molecular docking was used to determine whether 6-SH could spontaneously bind to death-associated protein kinase 1 (DAPK1). SPF-grade male SD rats were randomly divided into a sham group (n = 5), a CPR group (n = 7), and a CPR+6-SH group (n = 7). The CPR group and CPR+6-SH group were further divided into 12-, 24-, and 48-hour subgroups based on observation time points. A rat model of global CIRI after CA-CPR was established by asphyxiation. In the sham group, only tracheal and vascular intubation was performed without asphyxia and CPR induction. The CPR group was intraperitoneally injected with 1 mL of normal saline immediately after successful modeling. The CPR+6-SH group received an intraperitoneal injection of 20 mg/kg 6-SH (1 mL) immediately after successful modeling, followed by administration every 12 hours until the endpoint. Neurological Deficit Score (NDS) was recorded at each time point after modeling. After completion of observation at each time point, rats were anesthetized and sacrificed, and brain tissue specimens were collected. Histopathological changes of neurons were observed under light microscopy after hematoxylin-eosin (HE) staining. Ultrastructural changes of hippocampal neurons and autophagy were observed by transmission electron microscopy (TEM). Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect mRNA expression levels of DAPK1, vacuolar protein sorting 34 (VPS34), Beclin1, and microtubule-associated protein 1 light chain 3 (LC3) in brain tissues. Western blotting was used to detect protein expression levels of DAPK1, phosphorylated DAPK1 at serine 308 (p-DAPK1 ser308), VPS34, Beclin1, and LC3. Immunofluorescence was used to observe Beclin1 and LC3 expression in brain tissues under a fluorescence microscope.
RESULTS:
Molecular docking results indicated that 6-SH could spontaneously bind to DAPK1. Compared with the sham group, the NDS scores of the CPR group rats were significantly increased at all modeling time points; under light microscopy, disordered cell arrangement, widened intercellular spaces, and edema were observed in brain tissues, with pyknotic and necrotic nuclei in some areas; under TEM, mitochondria were markedly swollen with intact membranes, dissolved matrix, reduced or disappeared cristae, vacuolization, and increased autophagosomes. Compared with the CPR group, the NDS scores of the CPR+6-SH group rats were significantly decreased at all modeling time points; under light microscopy, local neuronal edema and widened perinuclear space were observed; under TEM, mitochondria were mostly mildly swollen with intact membranes, fewer autophagosomes, and alleviated injury. RT-qPCR results showed that compared with the sham group, mRNA expression levels of DAPK1, VPS34, Beclin1, and LC3 in brain tissues were significantly upregulated in all CPR subgroups, with the most pronounced changes at 24 hours. Compared with the CPR group, the CPR+6-SH group showed significantly lower mRNA expression of the above indicators at each time point [24 hours post-modeling (relative expression): DAPK1 mRNA: 3.41±0.68 vs. 4.48±0.62; VPS34 mRNA: 3.63±0.49 vs. 4.66±1.18; Beclin1 mRNA: 3.08±0.49 vs. 4.04±0.22; LC3 mRNA: 2.60±0.36 vs. 3.67±0.62; all P < 0.05]. Western blotting results showed that compared with the sham group, the protein expression levels of DAPK1, VPS34, Beclin1, and LC3 in all CPR subgroups were significantly increased, while the expression of p-DAPK1 ser308 was significantly decreased, with the most pronounced changes observed in the CPR 24-hour subgroup. Compared with the CPR group, the CPR+6-SH subgroups exhibited significantly reduced protein expression of DAPK1, VPS34, Beclin1, and LC3 [24-hour post-modeling: DAPK1/β-actin: 1.88±0.22 vs. 2.47±0.22; VPS34/β-actin: 2.55±0.06 vs. 3.46±0.05; Beclin1/β-actin: 2.12±0.03 vs. 2.87±0.03; LC3/β-actin: 2.03±0.24 vs. 3.17±0.23; all P < 0.05]. Conversely, the expression of p-DAPK1 ser308 was significantly upregulated in the CPR+6-SH group compared to the CPR group [24-hour post-modeling: p-DAPK1 ser308/β-actin: 0.40±0.02 vs. 0.20±0.07, P < 0.05]. Under the fluorescence microscope, fluorescence intensities of Beclin1 and LC3 in the CPR 24-hour group were significantly higher than those in the sham 24-hour group; compared with the CPR 24-hour group, the CPR+6-SH 24-hour group showed significantly reduced fluorescence intensities of Beclin1 and LC3.
CONCLUSION
6-SH inhibited the expression of DAPK1, alleviated excessive autophagy after global CIRI following CA-CPR in rats, and exerted neuroprotective effects. The mechanism may be related to phosphorylation at the DAPK1 ser308 site.
Animals
;
Rats, Sprague-Dawley
;
Male
;
Rats
;
Cardiopulmonary Resuscitation
;
Autophagy/drug effects*
;
Heart Arrest/therapy*
;
Death-Associated Protein Kinases/metabolism*
;
Reperfusion Injury/metabolism*
;
Disease Models, Animal
;
Neuroprotective Agents/pharmacology*
;
Brain Ischemia/metabolism*
2.Role of regulated cell death in the development of gastric cancer
Yang YU ; Yun XU ; Jingyuan CAO ; Junling HAN ; Chenglin ZHOU
Chinese Journal of Clinical Medicine 2024;31(4):652-658
Gastric cancer is one of the most common malignant tumors worldwide,with its incidence and mortality rates ranking third among malignant tumors in China.Regulated cell death(RCD)is a type of cell death activated by signal transduction modules and closely connected to the progression and treatment of gastric cancer.Different types of RCD,comprising apoptosis,pyroptosis,ferroptosis,cuproptosis,and autophagy,not only aid in eliminating damaged cells,but also serve a crucial role in suppressing gastric cancer spread.This paper reviews different forms of RCD and their roles in the progression of gastric cancer,so as to provide reference for new diagnosis and treatment strategies of gastric cancer.
3.Compatibility Mechanism of Mineral Medicine Os Draconis in Bupleuri Radix-containing Tri-herbal Medicines Based on Supramolecular Systems
Zi XING ; Junling HOU ; Yifan ZHAO ; Liman XIAO ; Mengjia WEI ; Mengyuan YANG ; Lu YUN ; Yuanfei NIU ; Zhijie ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):191-198
ObjectiveBy starting with the combination of Os Draconis, Bupleuri Radix, and Ostreae Concha, the role of mineral medicine Os Draconis in the combination of the Bupleuri Radix-containing tri-herbal medicines was preliminarily explored from the perspective of supramolecular system formation. Method① The appearance and Tyndall phenomenon of single decoction of Os Draconis, Bupleuri Radix, and Ostreae Concha, as well as co-decoction of Bupleuri Radix-Os Draconis, Bupleuri Radix-Os Draconis-Ostreae Concha, and Bupleuri Radix-Ostreae Concha were observed, and the average particle size, dispersion coefficient, and Zeta potential of suspension particles in each decoction were determined. The micromorphology of supramolecular structures was observed by scanning electron microscope (SEM). ② The pH of different compatibility systems, liquid viscosity coefficient, liquid surface tension, freeze-dried powder yield rate, and other physical properties were determined, and the interaction of different compatibility systems was detected by infrared absorption spectroscopy (FTIR) and UV-visible spectrophotometry (UV-Vis). ③ The composition and content difference of different compatible systems were determined by high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS). ResultCompared with the single decoction, the co-decoction had more obvious turbidity and Tyndall phenomenon. The particles in the co-decoction suspension were smaller and more evenly distributed, and the Zeta potential was reduced, indicating a more stable system. Under SEM, Bupleuri Radix was irregularly lamellar, and Bupleuri Radix-Os Draconis and Bupleuri Radix-Os Draconis-Ostreae Concha were mainly spherical nanoparticles. Bupleuri Radix-Ostreae Concha was irregularly lamellar, with a small number of spherical nanoparticles. The pH of the single decoction of Bupleuri Radix and co-decoction increased, and the viscosity coefficient increased. The liquid surface tension decreased. The freeze-dried powder yield rate of the Bupleuri Radix-Os Draconis co-decoction was the highest, followed by Bupleuri Radix-Ostreae Concha decoction and Bupleuri Radix-Os Draconis-Ostreae Concha decoction, and the yield rate of Bupleuri Radix single decoction was the lowest. The main change of FTIR was the stretching vibration of -OH, and the co-decoction moved to the low-frequency direction obviously. UV-Vis showed that the maximum absorption occurred at 295.8 nm for all groups, and the absorption intensity was different (Bupleuri Radix-Os Draconis>Bupleuri Radix-Os Draconis-Ostreae Concha>Bupleuri Radix-Ostreae Concha>Bupleuri Radix). The components of Bupleuri Radix were used as the indexes, and the content of methanol extract determined by HPLC was higher than that of water extract, and the components of Bupleuri Radix single decoction were mainly saikosaponin a (SSa) and saikosaponin c (SSc), which were slightly higher after co-decoction compatibility. UPLC-Q-TOF-MS could identify 37 compounds in both single decoction and co-decoction. ConclusionThe combination of Bupleuri Radix, Os Draconis, and Ostreae Concha can form a smaller, more uniform, and stable nano-sized supramolecular system, which is conducive to the dissolution of the main components of Bupleuri Radix, and the Os Draconis contributes the most in this process.
4.Detection and analysis of clinical specimens of COVID-19 in Anhui province
Qingqing CHEN ; Jun HE ; Junling YUN ; Yonglin SHI ; Weiwei LI ; Yuan YUAN ; Yinglu GE ; Nan SA ; Meng WANG ; Zhuhui ZHANG ; Yong SUN
Chinese Journal of Experimental and Clinical Virology 2020;34(4):382-384
Objective:To examine 2019 novel coronavirus (2019-nCoV) RNA in clinical specimens of COVID-19 patients in Anhui province, and provide evidence for laboratory diagnosis of COVID-19 and risk assessment of clinical specimens.Methods:ORF1ab gene and N gene of 2019-nCoV were detected by real-time fluorescence RT-PCR in 466 clinical specimens of 197 COVID-19 cases. Chi-square test was used to analyze the differences in positive rates of specimens with clinical classification and time of onset.Results:The positive rates of 2019-nCoV in throat swab, sputum, serum, blood sample were 88.83%, 94.67%, 6.78% and 5.08%. The positive rate for 2019-nCoV RNA in throat swabs and sputum differed significantly ( χ2=8.994, P=0.003) in common cases during 7 days after illness onset. Conclusions:The positive rate of RNA in sputum was higher than throat swabs. 2019-nCoV RNA was detected in serum and blood specimens of COVID-19 cases. There was a risk of serum and blood specimens for transmission of COVID-19.
5.Clinicopathologic study of 25 cases of placental chorioangioma.
Aichun WANG ; Yaqi MA ; Yun WANG ; Yingnan WANG ; Junling XIE ; Li WANG ; Jing YUAN ; Yiqun GU ; Aijun LIU ; E-mail: ALIU301@126.COM.
Chinese Journal of Pathology 2015;44(8):600-602
OBJECTIVETo investigate the clinicopathological characteristics, diagnostic criteria and differential diagnosis of placental chorioangioma.
METHODSTwenty-five cases of placental chorioangioma were analyzed for their clinical data, histomorphology and immumohisto chemical staining. Relevant literature was reviewed.
RESULTSThe average age of the 25 patients was 29 years. Fourteen patients had full-term pregnancy, 10 had preterm labor, and 1 had intrauterine fetal death. Nineteen patients had pregnancy complications. The tumors presented as red or dusty pink nodules with clear borders. The tumor size ranged from 1 to 16 cm. Microscopically, the tumors possessed abundant capillaries or cavernous blood spaces lined by hyperplastic endothelial cells. These cells were positive for CD34 and Ki-67 index < 10%.
CONCLUSIONSPlacental chorioangioma is a rare benign tumor of the placenta, and is associated with various pregnancy complications. Misdiagnosis of cell-rich type tumor should be avoided.
Adult ; Diagnosis, Differential ; Endothelial Cells ; pathology ; Female ; Fetal Death ; Hemangioma ; pathology ; Humans ; Infant, Newborn ; Placenta ; pathology ; Placenta Diseases ; pathology ; Pregnancy ; Pregnancy Complications, Neoplastic ; pathology ; Stillbirth
6.Clinical Research of EGFR and KRAS Mutation in Fine Needle Aspiration Cytology Specimens of Non-small Cell Lung Carcinoma
ZHANG ZHIHUI ; WU XILAN ; YING JIANMING ; LI JUNLING ; QIU TIAN ; GUO HUIQIN ; ZHAO HUAN ; SHAN LING ; LING YUN
Chinese Journal of Lung Cancer 2015;18(4):199-205
Background and objectiveEpidermal growth factor receptor (EGFR) and KARS must be detected mu-tation status before patients of lung cancer use targeted drugs. The aim of this study is to elucidate the signiifcance ofEGFR and KARS mutation in ifne needle aspiration (FNA) cytology suspension specimens of non-small cell lung carcinoma.Methods EGFR gene exons 18-21 andKARS codons 12, 13 of exons 2 were performed by Real-time PCR methods in ifne needle aspira-tion cytology suspension specimens of lymph nodes.Results 85 metastasis lymph nodes were detected in ifne needle aspira-tion cytology samples of lung cancer.EGFR mutation rate was 37.3%.KARS mutation rate was 7.2%. 19 formalin ifxed paraffn-embedded tissue specimens were available and match cytology specimens. Analysis ofEGFRmutation status in those samples revealed agreement with the results obtained in cytological samples (kappa=1.0). Clinical follow-up was available for 13 who presented with stage IV disease. Based on the identiifcation of such mutations, these patients received subsequent therapy with a TKI in clinic. We observed two cases complete remission (16.7%) and 8 cases partial remission (66.7%) and three had ongo-ing stable disease.ConclusionFine-needle aspiration cytology samples were detectedEGFR andKARS mutation. The meth-od which collects samples was easier, simple and convenient. This method has higher application value in clinical treatment.
7.Clinicopathologic study of 25 cases of placental chorioangioma
Aichun WANG ; Yaqi MA ; Yun WANG ; Yingnan WANG ; Junling XIE ; Li WANG ; Jing YUAN ; Yiqun GU ; Aijun LIU
Chinese Journal of Pathology 2015;(8):600-602
Objective To investigate the clinicopathological characteristics , diagnostic criteria and differential diagnosis of placental chorioangioma.Methods Twenty-five cases of placental chorioangioma were analyzed for their clinical data , histomorphology and immumohistochemical staining .Relevant literature was reviewed.Results The average age of the 25 patients was 29 years.Fourteen patients had full-term pregnancy , 10 had preterm labor , and 1 had intrauterine fetal death .Nineteen patients had pregnancy complications.The tumors presented as red or dusty pink nodules with clear borders.The tumor size ranged from 1 to 16 cm.Microscopically , the tumors possessed abundant capillaries or cavernous blood spaces lined by hyperplastic endothelial cells.These cells were positive for CD 34 and Ki-67 index <10%.Conclusions Placental chorioangioma is a rare benign tumor of the placenta , and is associated with various pregnancy complications .Misdiagnosis of cell-rich type tumor should be avoided .

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