Objective: To explore the short and mid-term efficacy of device closure of patent foramen ovale (PFO) for treating the patients with PFO combining cryptogenic stroke (CS) and transient ischemic attack (TIA). Methods: A total of 56 PFO patients with CS and TIA receiving device closure in our hospital from 2009-05 to 2015-12 were retrospectively studied. Transthoracic echocardiography (TTE), electrocardiogram (ECG), chest X-ray were examined at 24h, 1 month, 3 and 6 months after theoperation; telephone visit was conducted every 6 months thereafter. Results: There were 54/56 PFO patients combining CS and 2 combining TIA; 53 (94.6%)patients received PFO occluder from Starway medical technology. Aspirin was used for 6 months after the operation. The patients were followed-up for the average of (34.67±23.24) months. No body suffered from post-operative stroke and TIA; no residual shunt was observed. Conclusion: The short and mid-term efficacy of device closure has been satisfactory for treating the patients with PFO combining CS and TIA; its overall clinical value should be further investigated in large population and long-term study.

Objective To investigate the effect of low salt diet on vascular remodeling of rat induced by high fructose(HF).Methods Wistar male rats weighed 180-200 g were fed for 8 weeks and randomly divided into 6 groups:(1) control group was given the normal fodder and distilled water;(2) high fructose group(HF) was given normal fodder (0.5 ％ NaCl,w/w) and fructose water(10 ％,w/v);(3) high-salt group (HNa) was given high salt fodder (7 ％ NaCl,w/w) and distilled water;(4) high fructose combined with high salt diet group(HFNa) was simultaneously given high salt fodder and 10 ％ fructose water;(5)high fructose combined low salt group(HFLNa) was simultaneously given low salt fodder and 10％ fructose water;(6) high fructose combined with spirotaclone group(HFE) was given 10％ fructose water for 4 weeks and then added with spirotaelone(50 mg · kg-1 · d-1 by tube feeding) for continuous 4 weeks.The changes of arterial blood pressure,vascular wall histological evaluation and expression of α-SMA and fibronectin in vascular wall were detected in each group.Results (1) Compared with the blood pressure[(111.03 ±9.17) mm Hg] in the control group,the blood pressure in the HF and HNa groups were (133.94± 5.86) mm Hg and (128.09±7.56) mm Hg respectively,which were significantly increased(P＜0.05);(2) HF mainly caused the hyperplasia of vascular wall middle layer smooth muscle.The a-SMA expression results in the HF group was (0.006 3 ±0.000 21),which in the control group was (0.004 6 ± 0.000 31),the difference was statistically significant(P＜0.05),moreover which promoted the elastic fibers increase;while HNa mainly stimulated the elastic fibers to thicken and extracellular matrix deposition,the fibronectin expression was 0.002 6 ± 0.000 2 in the HNa group and (0.004 7±0.000 2)in the HF group,compared with(0.001 3±0.000 1)in the normal group,which were significantly increased(P＜0.001);(3) the blood pressure was (106.04±9.59) mm Hg in the HFLNa group,(103.99±7.12) mm Hg in the HFE group,compared with(133.94±5.86) mm Hg in the HF group,showing that the blood pressure in the HFLNa group and HFE group was significantly decreased compared with the HF group (P＜0.05);moreover the vascular remodeling in the HFLNa group(0.006 8±0.000 2) and HFE group (0.004 2±0.000 4) was improved,and compared with the HF group(0.006 3±0.000 2),α-SMA expression was significantly decreased (P＜0.05).Conclusion Low salt diet can effectively improve vascular remodeling induced by HEF.

Objective:To evaluate the relationship between milk fat globular epidermal growth factor Ⅷ (MFG-E8)-mediated efferocytosis and sepsis-associated encephalopathy (SAE) in mice.Methods:Forty clean-grade healthy male C57BL/6 mice, aged 2-3 months, weighing 18-22 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group Sham), cecal ligation and perforation (CLP) group (group CLP), sham operation+ phosphate buffer solution (PBS) group (group Sham+ PBS) and CLP+ recombinant mouse MFG-E8 group (group CLP+ rmMFG-E8). SAE was induced by CLP in anesthetized mice.PBS 1 μl and rmMFG-E8 1 μg were injected into the lateral cerebral ventricle in Sham+ PBS and CLP+ rmMFG-E8 groups after operation for 5 consecutive days.Novel object recognition test was performed at 6 days after operation, and the contextual fear conditioning test was performed at 7 and 8 days after operation.The percentage of time spent exploring a novel object, discrimination index and percentage of freezing time induced by condition were calculated.The animals were sacrificed after the end of behavioral tests, and the hippocampus was extracted for determination of the expression of MFG-E8 and GTP-Rac1 (by Western blot), the mRNA expression levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α and IL-1β (using real-time reverse transcription-polymerase chain reaction) and the apoptosis rate in hippocampus (by TUNEL). Results:Compared with group Sham, the percentage of time spent exploring a novel object, discrimination index and percentage of freezing time were significantly decreased, hippocamcal MFG-E8 expression was down-regulated, GTP-Rac1 expression was up-regulated, mRNA expression of IL-6, TNF-α and IL-1β was up-regulated, and apoptosis rate was increased in group CLP ( P<0.05). Compared with group CLP, the percentage of time spent exploring a novel object and discrimination index were significantly increased, freezing time was prolonged, hippocamcal MFG-E8 and GTP-Rac1 expression was up-regulated, mRNA expression of IL-6, TNF-α and IL-1β was down-regulated, and apoptosis rate was decreased in group CLP+ rmMFG-E8 ( P<0.05). Conclusion:The reduction of hippocampal MFG-E8-mediated efferocytosis may be involved in the pathophysiological mechanism of SAE in mice.

OBJECTIVE: To investigate the effects of gene of phosphate and tension homology (PTEN)-induced putative kinase 1 (PINK1)/Parkin pathway on hippocampal mitophagy and cognitive function in mice with sepsis-associated encephalopathy (SAE) and its possible mechanism. METHODS: A total of 80 male C57BL/6J mice were randomly divided into Sham group, cecal ligation puncture (CLP) group, PINK1 plasmid transfection pretreatment groups (p-PINK1+Sham group, p-PINK1+CLP group), empty vector plasmid transfection control group (p-vector+CLP group), with 16 mice in each group. The mice in CLP groups were treated with CLP to reproduce SAE models. The mice in the Sham groups were performed laparotomy only. Animals in the p-PINK1+Sham and p-PINK1+CLP groups were transfected with PINK1 plasmid through the lateral ventricle at 24 hours before surgery, while mice in the p-vector+CLP group were transfected with the empty plasmid. Morris water maze experiment was performed 7 days after CLP. The hippocampal tissues were collected, the pathological changes were observed under a light microscope after hematoxylin-eosin (HE) staining, and the mitochondrial autophagy was observed under a transmission electron microscopy after uranyl acetate and lead citrate staining. The expressions of PINK1, Parkin, Beclin1, interleukins (IL-6, IL-1β) and microtubule-associated protein 1 light chain 3 (LC3) were detected by Western blotting. RESULTS: Compared with the Sham group, CLP group mice in Morris water maze experiment had longer escape latency, shorter target quadrant residence time, and fewer times of crossing the platform at 1-4 days. Under the light microscope, the hippocampal structure of the mouse was injured, the neuronal cells were arranged in disorder, and the nuclei were pyknotic. Under the electron microscope, the mitochondria appeared swollen, round, and wrapped by bilayer or multilayer membrane structures. Compared with the Sham group, CLP group had higher expressions of PINK1, Parkin, Beclin1, LC3II/LC3I ratio, IL-6 and IL-1β in hippocampus, indicating that sepsis induced by CLP could activated inflammatory response and caused PINK1/Parkin-mediated mitophagy. Compared with the CLP group, p-PINK1+CLP group had shorter escape latencies, spent more time in the target quadrant and had more number of crossings in the target quadrant at 1-4 days. Under the light microscope, the hippocampal structures of mice was destroyed, the neurons were arranged disorderly, and the nuclei were pyknotic. Under transmission electron microscope, swollen and rounded mitochondria and mitochondrial structure wrapped by double membrane or multilayer membrane structure were observed. Compared with the CLP group, the levels of PINK1, Parkin, Beclin1 and LC3II/LC3 ratio in the p-PINK1+CLP group were significantly increased [PINK1 protein (PINK1/β-actin): 1.95±0.17 vs. 1.74±0.15, Parkin protein (Parkin/β-actin): 2.06±0.11 vs. 1.78±0.12, Beclin1 protein (Beclin1/β-actin): 2.11±0.12 vs. 1.67±0.10, LC3II/LC3I ratio: 3.63±0.12 vs. 2.27±0.10, all P < 0.05], while the levels of IL-6 and IL-1β were significantly decreased [IL-6 protein (IL-6/β-actin): 1.69±0.09 vs. 2.00±0.11, IL-1β protein (IL-1β/β-actin): 1.11±0.12 vs. 1.65±0.12, both P < 0.05], suggesting that overexpression of PINK1 protein could further activate mitophagy and reduce the inflammatory response caused by sepsis. There was no statistically significant difference in the above pathological changes and related indicators between Sham group and p-PINK1+Sham group, CLP group and p-vector+CLP group. CONCLUSIONS PINK1 overexpression can further activate CLP-induced mitophagy by upregulating Parkin, thereby inhibiting inflammation response and alleviate cognitive function impairment in SAE mice.
Male ; Animals ; Mice ; Mice, Inbred C57BL ; Sepsis-Associated Encephalopathy ; Phosphates ; Actins ; Beclin-1 ; Interleukin-6 ; Autophagy ; Ubiquitin-Protein Ligases ; Cognitive Dysfunction ; Sepsis ; Mitochondria ; Protein Kinases

Sepsis associated encephalopathy (SAE) is a severe disease secondary to sepsis, which is associated with increased mortality and causes long-term cognitive deficits in survivors. Recently, an increasing body of evidence has shown that gut microbiota is closely related to the central nervous system, and could influence brain function via microbiota-gut-brain axis. Therefore, in the occurrence and development of SAE, cholinergic anti-inflammatory pathway is one of the mechanisms by which gut microbiota could improve cognitive function. Efferocytosis, a process of eliminating apoptotic cells in the body, has anti-inflammatory effects and provides organ protection in sepsis. On the other hand, it could be enhanced by some metabolites of gut microbiota, making it another potential mechanism for gut microbiota regulating SAE. This review summarizes the mutual regulation of gut microbiota, efferocytosis and SAE, to explore potential mechanisms and therapeutic targets of SAE.

Objective:To explore the potential mechanism of PD-1 inhibitor P on RIMI from the perspective of immune microenvironment.Methods:To establish a mouse model of radiation-induced myocardial injury (RIMI), twenty C57BL/6 mice were randomly divided into 4 groups, 5 in each group. Group A was the healthy control group; Group B was the PD-1 inhibitor group; Group C was the simple irradiation group, with a heart irradiation of 15 Gy; Group D was the irradiation+ PD-1 inhibitor group. One month after irradiation, the mice were anesthetized and sacrificed. The morphological changes of myocardial tissues were observed by HE staining. The myocardial fibrosis was assessed by Masson staining. CD 3+ , CD 3+ CD 4+ , CD 3+ CD 8 lymphocyte subsets and cytokines (IL-4, IL-6, IL-17A, TNF-α, TGF-β 1 and INF-γ) levels were determined by flow cytometry. The apoptosis rate of myocardial cells was detected by TUNE. Results:One month after irradiation, there was no obvious myocardial fibrosis in group B, and collagen fibers were distributed in the interstitium of myocardial cells in groups C and D. Semi-quantitative analysis results showed that the myocardial collagen volume fraction (CVF) of groups A, B, C and D were (1.97±0.36)%, (2.83±1.03)%, (5.39±0.77)% and (7.72±1.43)%, respectively. The CVF between group A and group B was similar ( P=0.314), and the differences in CVF between the other groups were statistically significant (all P<0.05). Compared with group A, the absolute value and percentage of CD 3+ T lymphocytes were significantly increased in groups B, C and D (all P<0.01). The values in group D were significantly higher than those in group B and group C (all P<0.01); The absolute value and percentage of CD 3+ CD4 T lymphocytes were similar among four groups (all P>0.05); The absolute value and percentage of CD 3+ CD 8 T lymphocytes in group D were significantly higher than those in groups A, B and C (all P<0.001). The expression levels of IL-6, IL-17A, and TGF-β 1 in group D were significantly higher compared with those in groups A, B and C (all P<0.001). The apoptotic index was gradually increased in four groups, and the differences in apoptotic index among four groups were statistically significant (all P<0.001). Conclusion:PD-1 inhibitors can aggravate RIMI by promoting myocardial immune inflammatory response.