Study on the hepatotoxicity and potential mechanism of gefitinib based on CYP450 in mice and AML12 cells.

OBJECTIVES: Gefitinib is mainly used for the treatment of non-small-cell lung cancer. Hepatotoxicity is one of the main side effects of gefitinib, and seriously affects the treatment process of the disease. However, the hepatotoxicity mechanism of gefitinib remains unclear. METHODS: The hepatotoxicity of different doses of gefitinib was investigated in mice and AML-12 cells, and the possible correlation of hepatotoxicity with CYP450 was analysed. KEY FINDINGS: The toxic effects of gefitinib were confirmed by the increased liver index, decreased body weight and survival rate, injured liver function and histopathology followed 16 days of oral administration. Gefitinib (400 mg/kg) upregulated the hepatic mRNA expression of CYP1A1 and downregulated the CYP2D9 and CYP2D10 in mice. Furthermore, we verified that gefitinib produced cytotoxicity on AML-12 cells in a dose and time-dependent manner, and confirmed that gefitinib (20 µM) induced cell apoptosis, upregulated mRNA expression of CYP1A1 and downregulated CYP2D9 and CYP2D10. Pearson correlation analysis also showed that the hepatotoxicity of gefitinib was positively correlated with CYP1A1 and negatively correlated with CYP2D9 and CYP2D10. CONCLUSIONS: Our results suggested that the hepatotoxicity gefitinib may be associated with CYP1A1, CYP2D9 and CYP2D10. These findings will contribute to a better understanding of the mechanism of gefitinib hepatotoxicity.

The antidepressant-like effect of formononetin on chronic corticosterone-treated mice.

Previous studies reported the neuroprotective effects of formononetin (FMN), however, whether it has antidepressant-like effects have not been reported. To evaluate the antidepressant-like effects of FMN, a mice model of depression was established by chronic corticosterone (CORT) injection. The serum corticosterone levels and hippocampal protein expression were detected by ELISA and Western blot. Nissl staining was used to observe the damage of hippocampal neurons and immunofluorescence was used to observe the neurogenesis in the hippocampus. Our results showed that FMN significantly increased the sucrose preference and shorten the immobility time in the forced swimming test in CORT-treated mice. Moreover, FMN reduced the serum corticosterone levels, upregulated the protein expression levels of the glucocorticoid receptor (GR), and brain-derived neurotrophic factor (BDNF) in the hippocampus, protected against the CORT-induced neuronal impairment, and promoted the neurogenesis in the hippocampus. Taken together, the present study was the first to demonstrate the antidepressant-like effects of FMN in the CORT-induced mice model of depression, which may contribute to the discovery of a new candidate for treating depression.

Tandem mass tag-based quantitative proteomic analysis of the liver reveals potential protein targets of Xiaochaihutang in CUMS model of depression.

Depression is a global mental disorder disease and greatly threatened human health. Xiaochaihutang (XCHT) has been used successfully in treatment of depression for many years in China, but the mechanism is unclear. Using the chronic unpredictable mild stress (CUMS) mice model of depression, the present study aimed to reveal possible antidepressant mechanisms of XCHT from the perspective of liver by analyzing hepatic proteomics in mice. Bioinformatics analysis identified 31 differentially expressed proteins (DEPs), including 5 upregulated and 26 downregulated proteins, between the CUMS model and XCHT groups. The bile secretion pathway was found by KEGG pathway analysis of these DEPs. Four of the 31 differentially expressed proteins, including 2 active proteins involved in bile secretion, carbonic anhydrase 2 (CA2) and cystic fibrosis transmembrane conductance regulator (CFTR), were selected to verify their genes. Four genes (Cyp7a1, Fxr, Shp and Ntcp) related to bile acid synthesis and transport were further investigated by quantitative real-time polymerase chain reaction (qRT-PCR). Both biochemical tests and gene studies demonstrated that CUMS affected bile acid synthesis and transport, while XCHT regulated this pathway. The results indicated that there may be a potential relationship between CUMS induced depression and hepatic injury caused by increased bile acid, and also provide a novel insight to understand the underlying anti-depression mechanisms of XCHT.

Fulminant myocarditis induced by immune checkpoint inhibitor nivolumab: a case report and review of the literature.

BACKGROUND: Nivolumab, an anti-programmed cell death protein 1 antibody, is commonly used as an immune checkpoint inhibitor in various cancers. Various adverse events are associated with these therapies, including hepatitis, dermatitis, and myocarditis. Myocarditis is a relatively rare but potentially fatal immune-mediated adverse reaction. CASE PRESENTATION: We report a case of colon cancer in a 56-year-old Chinese patient with lung and liver metastasis who developed fulminant myocarditis by nivolumab and survived with the support of extracorporeal membrane oxygenation. After six cycles (within 3 months) of nivolumab treatment, the patient developed chest tightness and was hospitalized. A diagnosis of fulminant myocarditis associated with immunotherapy was confirmed based on the clinical manifestations and laboratory examinations. He recovered well and was discharged on day 45 after management with extracorporeal membrane oxygenation, intravenous methylprednisolone, and immunoglobulin. CONCLUSIONS: This case illustrates a severe cardiovascular complication of immunotherapy, strongly suggesting the necessity of close monitoring for outpatient usage of nivolumab. Additionally, our experience provided an efficient management strategy of extracorporeal membrane oxygenation in terms of life-threatening conditions.

Quantitative proteomic analysis of the liver reveals antidepressant potential protein targets of Sinisan in a mouse CUMS model of depression.

In traditional Chinese medicine, the role of the liver in depression is highly valued, and liver-relieving drugs, such as Sinisan, are often used to treat depression; however, the mechanism whereby these drugs work remains unclear. The present study aimed to reveal possible antidepressant mechanisms of Sinisan (SNS) by analyzing hepatic proteomics in chronic unpredictable mild stress (CUMS) mice. Using the CUMS mouse model of depression, the antidepressant effects of SNS were assessed by the sucrose preference test (SPT) and forced swimming test (FST). Hepatic differentially expressed proteins (DEPs) after SNS treatment were investigated by tandem mass tag (TMT) based quantitative proteomics analysis. Then, a bioinformatics analysis of DEPs was conducted through hierarchical clustering, Venn analysis, Gene Ontology (GO) annotation enrichment, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. DEP genes were further validated by quantitative real-time polymerase chain reaction (qRT-PCR) analysis and western blotting. Behavioral results demonstrated that SNS significantly increased sucrose intake in SPT and shortened the immobility time in FST in model mice. Eighty-two DEPs were identified, including 37 upregulated and 45 downregulated proteins, between model and SNS groups. Enrichment analysis of GO annotations indicated that SNS primarily maintained cellular iron ion homeostasis by iron ion transportation and regulated expression of some extracellular structural proteins for oxidation-reduction processes. KEGG and Venn analysis showed that mineral absorption, steroid hormone biosynthesis and metabolism might be the principal pathways through which SNS acts on depression. Furthermore, several proteins involved in the biosynthesis and metabolism of steroid hormone pathways were significantly up/downregulated by SNS, including CYP2B19, CYP7B1 (validated by qRT-PCR) and HSD3b5 (validated by qRT-PCR and western blotting). Our results indicate that SNS plays important roles in antidepressant actions by restoring DEPs, resulting in the biosynthesis and metabolism of steroid hormones. The current results provide novel perspectives for revealing potential protein targets of SNS in depression.

[Effects of mechanical ventilation on diaphragmatic contractile function in patients with AECOPD].

OBJECTIVE: To study diaphragmatic strength in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) during mechanical ventilation (MV), and to explore the predictive value of maximal transdiaphragm pressure (Pdi max) for successful extubation. METHODS: A prospective study was conducted. Twenty-one patients with AECOPD receiving MV admitted to intensive care unit (ICU) of Shanghai East Hospital of Tongji University of School Medicine from February 2015 to May 2017 were enrolled. Pdi max value was measured by using esophageal and gastric balloon catheters within 24 hours of intubation until the day the patient underwent extubation or died. In addition, the C-reactive protein (CRP), serum albumin (Alb) and prealbumin (PA) during MV were recorded. Pearson correlation was used to analyze the correlations between Pdi max and CRP, Alb and PA. The receiver operating characteristic curve (ROC) was used to cumulate Pdi max value of the successful weaning. RESULTS: Tracheotomy was done in 2 patients, and 2 patients quit the study. The remaining 17 patients were included in the investigative protocol. Six of the 17 patients died and 11 patients were successfully extubated. (1) Mean Pdi max decreased progressively over time in 17 patients of AECOPD during MV. There were no significant changes in Pdi max at 1-2 days of MV. Mean Pdi max at 7 days was significantly lower than that at 3 days [cmH2O (1 cmH2O = 0.098 kPa): 20.2±4.2 vs. 28.1±4.4, P < 0.01]. By the end of the evaluation period at 11 days of MV, mean Pdi max decreased about 38.7% to the 1 day of MV (cmH2O: 19.8±4.7 vs. 32.3±3.9, P < 0.01). During MV, CRP, Alb and PA showed a downward trend. (2) Mean Pdi max and the Pdi max before extubation in patients with difficulty extubation from MV was lower than that in successful weaning [Mean Pdi max (cmH2O): 25.2±5.4 vs. 28.9±5.8, Pdi max before extubation (cmH2O): 16.9±2.8 vs. 26.8±6.6, both P < 0.01]. (3) There was significantly negative correlation between Pdi max value and CRP (r = -0.799, P = 0.000). There was significantly positive correlation between Pdi max value and serum Alb (r = 0.613, P = 0.008) and PA (r = 0.661, P = 0.004). (4) ROC curve analysis showed that the area under the ROC curve (AUC) for predicting weaning success in the patients with AECOPD was 0.902. The sensitivity of the diagnosis was 81.8% and the specificity was 100% when cut-off value of Pdi max was 23.2 cmH2O. CONCLUSIONS: MV induced the reduction of diaphragmatic contractility in a time-dependent manner. The Pdi max in patients with difficult extubation from MV was lower than that in successful weaning. The Pdi max could be a parameter to predict the successful extubation in patients with AECOPD during MV.

[A study of lymphocyte apoptosis and endoplasmic reticulum stress in the development of sepsis and their association with outcome in septic patients].

OBJECTIVE: To investigate the role of lymphocyte apoptosis and endoplasmic reticulum stress (ERS) on the development of sepsis and their association with the prognosis of sepsis patients. METHODS: A prospective cohort study was conducted. Seventy septic patients admitted to intensive care unit (ICU) of Shanghai East Hospital of Tongji University were enrolled. Blood samples were collected on days 1, 3 and 7 to measure percentage of circulating apoptotic lymphocyte with flow cytometry analysis. The relative expressions of endoplasmic reticulum specific glucose regulated protein 78 (GRP78) mRNA and transcription factor CHOP mRNA were measured by real-time reverse transcription-polymerase chain reaction (RT-PCR). The correlation between CHOP mRNA expression and percentage of circulating apoptotic lymphocyte was analyzed by Spearman relative analysis. The patients were divided into death (n = 23) and survival subgroups (n = 47). Twenty healthy volunteers during the same period were selected as the healthy control group. RESULTS: (1) Rate of lymphocyte apoptosis: compared with healthy control group [(2.86±0.66)%], septic patients, either survival or death subgroup, exhibited higher rate of lymphocyte apoptosis on days 1, 3 and 7 [survival subgroup: (12.44±4.43)%, (8.57±3.38)%, (6.78±3.35)%; death subgroup: (14.42±2.01)%, (11.32±2.53)%, (8.87±3.62)%, all P < 0.01], and it was obvious on day 1, and the phenomenon became less marked gradually. The rate of circulating apoptotic lymphocytes did not differ between the death and survival subgroups on day 1, but there was a significant difference in the rate on day 3 and day 7 (both P < 0.05). (2) The expression of CHOP mRNA (2(-ΔΔCt)): compared with that in healthy controls [(2.56±1.09)×10-3], CHOP mRNA expression was increased on days 1, 3 and 7 in septic patients [survival subgroup: (5.83±1.96)×10(-3), (4.24±1.60)×10(-3), (4.15±1.64)×10(-3), death subgroup: (37.20±20.70)×10(-3), (18.80±13.90)×10(-3), (9.28±7.78)×10(-3), all P < 0.01], and it was more obvious in the death subgroup, as it was increased by 6.38, 4.43, and 2.24 folds (P values was 0.000, 0.000, and 0.001), but it decreased rapidly in death subgroup. (3) The expression of GRP78 mRNA (2(-ΔΔCt)): compared with healthy controls [(3.31±2.04)×10(-3)], the expression of GRP78 mRNA in both survival and death subgroups increased in septic patients on day 1 [(5.83±2.00)×10-3, (11.30±6.48)×10(-3), both P < 0.01], and they decreased subsequently. The expression of GRP78 mRNA in the survival subgroup declined to the levels of the healthy control group on day 3 and day 7 [3 days: (3.99±1.60)×10(-3), 7 days: (3.30±1.35)×10(-3), both P > 0.05], and GRP78 mRNA expression in the death subgroup was gradually lowered, but it was still higher than that in the healthy control group [3 days: (7.27±3.64)×10(-3), 7 days: (5.23±1.94)×10(-3), both P < 0.01]. (4) Spearman relative analysis showed that the expression of CHOP mRNA was positively correlated with the rate of lymphocyte apoptosis (r = 0.414, P = 0.000). CONCLUSIONS: The increase in the rate of lymphocyte apoptosis and activation of ERS play an important role in the development of sepsis, and it is associated with worse outcome in the septic patients.

Ginsenoside Rb1 improves postoperative fatigue syndrome by reducing skeletal muscle oxidative stress through activation of the PI3K/Akt/Nrf2 pathway in aged rats.

Ginsenoside Rb1 is reported to possess anti-fatigue activity, but the mechanisms remain unknown. The aim of this study was to investigate the molecular mechanisms responsible for the anti-fatigue effect of ginsenoside Rb1 on postoperative fatigue syndrome induced by major small intestinal resection (MSIR) in aged rat. Aged rats with MSIR were administrated with ginsenoside Rb1 (15 mg/kg) once a day from 3 days before surgery to the day of sacrifice, or with saline as corresponding controls. Rats without MSIR but going through the same surgery procedure were administrated with saline as blank controls. Anti-fatigue effect was assessed by an open field test; superoxide dismutase, reactive oxygen species and malondialdehyde in skeletal muscle were determined. The mRNA levels of Akt2 and Nrf2 in skeletal muscle were measured by real-time quantitative PCR. The activation of Akt and Nrf2 was examined by western blot and immunohistofluorescence. Our results revealed that ginsenoside Rb1 significantly increased the journey and the rearing frequency, decreased the time of rest in aged rats with MSIR. In addition, ginsenoside Rb1 significantly reduced reactive oxygen species and malondialdehyde release and increased the superoxide dismutase activity of skeletal muscle in aged rats with MSIR. Ginsenoside Rb1 also increased the expression of Akt2 and Nrf2 mRNA, up-regulated Akt phosphorylation and Nrf2 nuclear translocation. These findings indicate that ginsenoside Rb1 has an anti-fatigue effect on postoperative fatigue syndrome in aged rat, and the mechanism possibly involves activation of the PI3K/Akt pathway with subsequent Nrf2 nuclear translocation and induction of antioxidant enzymes.

Fast-track surgery in gastrectomy for gastric cancer: a systematic review and meta-analysis.

PURPOSE: Fast-track surgery aims to attenuate the surgical stress response, reduce complications, and shorten hospital stay. The goal of the present meta-analysis is to assess the safety and effectiveness of fast-track surgery in patients undergoing gastrectomy for gastric cancer compared with conventional perioperative care. METHODS: PubMed, Embase, the Cochrane Central Register of Controlled Trials, and reference lists of the identified studies were searched to identify randomized clinical trials that compared fast-track surgery with conventional perioperative care in patients undergoing gastrectomy for gastric cancer. RESULTS: Five studies with a total of 400 patients were included in the meta-analysis. Meta-analysis shows that postoperative hospital stay (weighted mean difference (WMD) -1.87 days, 95 % confidence interval (CI), -2.46 to -1.28 days, P < 0.00001), time to first passage of flatus (WMD -0.71 days, 95 % CI, -1.03 to -0.39 days, P < 0.0001), and hospital costs (WMD -505.87 dollars, 95 % CI, -649.91 to -361.84 dollars, P < 0.00001) were significantly reduced for fast-track surgery. No significant differences were found for readmission rates (relative risk (RR), 1.97 95 % CI, 0.37 to 10.64, P = 0.43) and total postoperative complications (RR, 0.99 95 % CI, 0.56 to 1.76, P = 0.97). CONCLUSIONS: Fast-track surgery is safe and effective in gastrectomy for gastric cancer. Further randomized trials are needed to strengthen the conclusions.

Early versus traditional postoperative oral feeding in patients undergoing elective colorectal surgery: a meta-analysis of randomized clinical trials.

BACKGROUND: The safety and effectiveness of early oral feeding after colorectal surgery has not been determined. We performed a meta-analysis to evaluate surgical outcomes following early oral feeding compared with traditional oral feeding in patients undergoing elective colorectal surgery. METHODS: MEDLINE, EMBASE, and the Cochrane Central Register of Controlled Trials were searched to identify randomized clinical trials comparing the outcomes following early oral feeding versus traditional oral feeding in patients undergoing elective colorectal surgery. The trials must have reported at least one of the following end points: anastomotic dehiscence, pneumonia, wound infection, nasogastric tube reinsertion, vomiting, mortality, length of hospital stay, hospital costs, and quality of life. RESULTS: Seven trials, which included a total of 587 patients, met our inclusion criteria. Compared with traditional oral feeding, early oral feeding reduced the length of hospital stay (weighted mean difference -1.58 days; 95% CI -2.77 to -0.39; p = 0.009) and the total postoperative complications (relative risk 0.70; 95% CI 0.50-0.98; p = 0.04). There were no significant differences in the risk of anastomotic dehiscence, pneumonia, wound infection, rate of nasogastric tube reinsertion, vomiting, or mortality. CONCLUSIONS: Early oral feeding is safe and effective in patients undergoing elective colorectal surgery.