Objective To investigate the mechanism of AMP-activated protein kinase (AMPK) in hepatic ischemia-reperfusion injury (HIRI). Methods ① Grouping. Forty-two mice were randomly divided into Sham group, 4 ischemia reperfusion (IR) group of different times (2, 6, 12, and 24 h), Compound C group, and Compound C+repamycin (Rapa) group, each group enrolled in 6 mice. Compound C group: mice were modeled at 1 h after intraperitoneal injection of Compound C (25 mg/kg). Compound C+Rapa group: mice were modeled at 1 h after intraperitoneal injection of rapamycin (1 mg/kg) and Compound C (25 mg/kg). Mice of 4 IR groups, Compound C group, and Compound C+Rapa group were used to prepare HIRI model. Mice of Sham group were treated only for laparotomy, freeing the first portal hepatis and closing peritoneal. ② To filter the best IR time. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum of mice in Sham group and IR groups of 4 different reperfusion time points were measured. The pathological changes of liver tissues were observed by HE staining, and the expressions of related proteins in liver tissue of mice were detected by Western blot. Considering the results of blood biochemical test, HE staining, and Western blot together to determine the best IR point. ③ The exploration of signal pathway for AMPK. The expressions of proliferating cell nuclear antigen (PCNA) were observed by immunohistochemical staining in the liver tissues of IR-12 h group, Compound C group (12 h after IR) and compound C+Rapa group (12 h after IR). The mitochondrial damage was observed by rhodamine 123 staining, and the apoptotic status of liver cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay (TUNEL). Results ① The 12 h after IR was the best observation time point. Compared with IR-12 h group, the levels of ALT and AST in Sham group, IR-2, 6, and 24 h groups were lower (P<0.05). HE staining showed that liver tissue destruction in IR-12 h group was the most severe. Western blot showed that, expressions of AMPKα, phosphorylated adenylate activated protein kinase α (p-AMPKα), Nip3-like protein X (Nix), BCL-2 homologous water-soluble protein (Bax), as well as ratio of autophagy microtubule-associated protein light chain 3 (LC3)Ⅱto LC3Ⅰof Sham group, IR-2, 6, and 24 h group were all lower than those of IR-12 h group (P<0.05), but the expressions of phosphorylated mammalian target of Rapa (p-mTOR) of Sham group, IR-2, 6, and 24 h group were all higher (P<0.05). Therefore, 12 h after IR was the best time to observe. ② Compared with IR-12 h group, the expression level of PCNA protein in liver tissue of Compound C group was lower (P<0.05), the mitochondrial luminescence intensity was weaker and the apoptotic cells were more. Compared with Compound C group, the expression of PCNA protein in the liver tissue of the Compound C+Rapa group was higher (P<0.05), the mitochondrial intensity was stronger and the apoptotic cells were less. ③ Compared with IR-12 h group, the expressions of Nix and p-AMPKα, and ratio of LC3Ⅱ to LC3Ⅰ in liver tissue of Compound C group decreased (P<0.05), while the expressions of p-mTOR, Caspase-3, and Cleaved Caspase-3 increased (P<0.05). Compared with Compound C group, the expressions of p-AMPKα and Nix in the liver tissue of Compound C+Rapa group increased (P<0.05), while the expressions of p-mTOR, Caspase-3, and Cleaved Caspase-3 decreased (P<0.05). Conclusion During the HIRI in mouse, AMPK regulates mitophagy and apoptosis through the mTOR/Nix pathway.
Objective To investigate the clinical features, treatment, and influence factors of prognosis in patients with gastric neuroendocrine neoplasms (GNENs). Methods From March 2011 to January 2016, the clinicopathological data of 44 patients with GNENs who treated in The Affiliated Hospital of Xuzhou Medical University were retrospectively analyzed to summarize the choice of treatment plan and analyze influence factors of prognosis. Results A total of 44 patients enrolled in this study. The major clinical manifestation included abdominal pain in 18 patients (40.9%), abdominal distension in 16 patients (36.4%), loss of appetite in 4 patients (9.1%), acid regurgitation and belching in 4 patients (9.1%), nausea and vomiting in 1 patient (2.3%), eating after choking sense in 3 patients (6.8%), gastrointestinal bleeding in 2 patients (4.5%), diarrhea in 1 patient (2.3%), and palpitations with weakness in 3 patients (6.8%). The treatment of 44 patients included radical resection in 26 patients (59.1%), endoscopic resection in 13 patients (29.6%), local excision in 1 patient (2.3%), and 4 patients had distant metastasis before operation were conducted of palliative treatment〔palliative resection in 2 patients (4.5%) and conservative treatment in 2 patients (4.5%)〕. Univariate analysis showed that the gender, the age, the tumor size, and the N staging (lymph node metastasis) were not associated with prognosis (P>0.050), but the tumor location and the depth of tumor invasion were related to the prognosis (P<0.050). The tumors located in the upper part of the stomach and the serosal infiltration indicated poor prognosis. However, neither of them can be used as independent factor to evaluate the poor prognosis of GNENs patients (P>0.050). Conclusions GNENs has nonspecific clinical manifestation. Radical surgery and endoscopic resection are the main treatment methods, but the influence factors of prognosis in GNENs patients need further study.