ObjectiveTo examine the influences of Shenfu injection on the endogenous metabolic byproducts in the myocardium of the rat model exhibiting chronic heart failure， thus deciphering the therapeutic mechanism of the Qi-reinforcing and Yang-warming method. MethodsSD rats were randomly allocated into a control group and a modeling group. Chronic heart failure with heart-Yang deficiency syndrome in rats was modeled by multi-point subcutaneous injection of isoproterenol， and the rats were fed for 14 days after modeling. The successfully modeled rats were randomized into model， Shenfu injection （6.0 mL·kg^-1）， and trimetazidine （10 mg·kg^-1） groups and treated with corresponding agents for 15 days. The control group and the model group were injected with equal doses of normal saline， and the samples were collected after the intervention was completed. Cardiac color ultrasound was performed. Hematoxylin-eosin （HE） staining was used to observe histopathological morphology， and the serum level of N-terminal pro-brain natriuretic peptide （NT-proBNP） was assessed by enzyme-linked immunosorbent assay （ELISA）. The mitochondrial morphological and structural changes of cardiomyocytes were observed by transmission electron microscopy， and the metabolic profiling was carried out by ultra high performance liquid chromatography-quantitative exactive-mass spectrometry （UHPLC-QE-MS）. Differential metabolites were screened and identified by orthogonal partial least squares-discriminant analysis （OPLS-DA） and other methods， and then the MetaboAnalyst database was used for further screening. The relevant biological pathways were obtained through pathway enrichment analysis. The receiver operating characteristic （ROC） curve was established to evaluate the diagnostic value of each potential biomarker for myocardial injury and the evaluation value for drug efficacy. ResultsThe results of color ultrasound showed that Shenfu Injection improved the cardiac function indexes of model rats （P<0.05）. The results of HE staining showed that Shenfu injection effectively alleviated the pathological phenomena such as myocardial tissue structure disorder and inflammatory cell infiltration in model rats. The results of ELISA showed that Shenfu injection effectively regulated the serum NT-proBNP level in the model rats. Transmission electron microscopy （TEM） showed that Shenfu injection effectively restored the mitochondrial morphological structure. The results of metabolomics showed that the metabolic phenotypes of myocardial samples presented markedly differences between groups. Nine differential metabolites could be significantly reversed in the Shenfu injection group， involving three metabolic pathways： pyruvate metabolism， histidine metabolism， and citric acid cycle （TCA cycle）. The results of ROC analysis showed that the area under the curve （AUC） values of all metabolites were between 0.75 and 1.0， indicating that the differential metabolites had high diagnostic accuracy for myocardial injury， and the changes in their expression levels could be used as potential markers for efficacy evaluation. ConclusionShenfu injection significantly alleviated the damage of cardiac function， myocardium， and mitochondrial structure in the rat model of chronic heart failure with heart-Yang deficiency syndrome by ameliorating energy metabolism remodeling. Reinforcing Qi and warming Yang is a key method for treating chronic heart failure with heart-Yang deficiency syndrome.

Objective:To evaluate the effect of sevoflurane postconditioning on early inflammatory responses during intestinal ischemia-reperfusion (I/R) in rats.Methods:Sixty clean-grade Sprague-Dawley rats, aged 8-10 weeks, weighing 200-230 g, were divided into 3 groups ( n=20 each) using a random number table method: sham operation group (Sham group), intestinal I/R group (I/R group), and sevoflurane postconditioning group (Sevo group). Intestinal I/R was produced by occlusion of the superior mesenteric artery for 60 min followed by reperfusion for 2 h in anesthetized rats in I/R group and Sevo group.Laparotomy was performed, and the superior mesenteric artery was only isolated in Sham group.The rats inhaled 1.15% sevoflurane for 30 min for postconditioning in Sevo group.Blood samples were collected by cardiac puncture at 2 h of reperfusion, and then the rats were sacrificed.Samples of intestine were obtained for examination of the pathological changes of intestinal tissues (with a light microscope) which were scored according to Chiu and for determination of the neutrophil L-selectin levels in blood (by flow cytometry), tumor necrosis factor-alpha (TNF-a) expression (by Western blot), myeloperoxidase (MPO) activity (by spectrophotometry). Results:Compared with group Sham, the neutropil L-selectin level in blood was significantly increased in group I/R and decreased in group Sevo, and Chiu′s score, TNF-a expression and MPO activity were significantly increased in I/R and Sevo groups ( P<0.05). Compared with group I/R, the neutropil L-selectin level in blood, Chiu′s score, TNF-a expression, TNF-a expression and MPO activity were significantly decreased ( P<0.05), and the pathological changes were significantly attenuated in group Sevo. Conclusion:The mechanism by which sevoflurane postconditioning reduces intestinal I/R injury may be related to inhibiting early inflammatory responses in rats.

Objective To explore the variation trend of natural killer (NK) cell subsets in the recipients infected with cytomegalovirus (CMV) after renal transplantation. Methods Clinical data of 92 renal transplant recipients were retrospectively analyzed. All recipients were divided into the CMV infection group (n=43), CMV infection recovery group (n=13), stable renal function group (n=15), rejection group (n=11) and other infection group (n=10). In addition, healthy adult volunteers were enrolled in the healthy control group (n=15). The proportion of NK cells in peripheral blood, the expression proportion and the mean fluorescence intensity (MFI) of CD226 and CD16 in NK cells were observed and statistically compared among different groups. Results The proportion of NK cells was 4.9% (2.2%, 11.5%) in the CMV infection group and 3.7% (2.3%, 6.5%) in the CMV infection recovery group, which were significantly lower than those in the other groups (all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection group was significantly lower compared with those in the healthy control group and stable renal function group(all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection recovery group was remarkably higher than those in the CMV infection group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection group was significantly lower than those in the healthy control group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection recovery group was significantly higher than those in the CMV infection group (both P < 0.05). Conclusions The expression proportion and MFI of CD226 and CD16 in NK cells are down-regulated in CMV infection period, whereas up-regulated during the CMV infection recovery period, prompting that CD226 and CD16 expressed by NK cells are intimately correlated with the course of CMV infection.

Objective To evaluate the effects of ketamine anesthesia on proteome in hippocampus in aged rats.Methods Thirty healthy male Wistar rats,aged 20 months,weighing 560-610 g,were di-vided into 2 groups(n＝15 each)using a random number table method: control group(group C)and ket-amine group(group K).In group K,ketamine 80 mg/kg was intraperitoneally injected,additional 1/2 ini-tial dose was given when the righting reflex was recovered,and anesthesia was maintained for 3 h.Morris water maze test was performed starting from 1st day after the end of anesthesia.Five rats were selected at days 1 and 7 after the end of anesthesia and sacrificed,and hippocampal tissues were obtained to extract proteins.Proteins extracted from rat hippocampi were identified by 2-dimensional electrophoresis(2-DE).The differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and biological information system.Results Compared with group C,the escape latency and total swimming distance to find the submerged platform in Morris water maze at the 1st day after anesthesia were significantly prolonged in group K(P＜0.05 or 0.01).The MAL-DI-TOF-MS analysis showed that there were 21 differentially expressed proteins at 1st day after ketamine an-esthesia,of which 6 proteins(involving maintenance of intracellular protein homeostasis,energy metabo-lism,etc.)presented with up-regulated expression and 15 proteins(involving synaptic vesicle transport ef-ficiency,synaptic structural and functional plasticity,maintenance of intracellular protein homeostasis,NMDA-mediated Ca2+signal transport,energy metabolism,etc.)presented with down-regulated expres-sion.There were 8 differentially expressed proteins at 7th day,including 3 proteins with up-regulated ex-pression and 5 proteins with down-regulated expression(P＜0.05).Conclusion Ketamine anesthesia can induce 21 differentially expressed proteins in hippocampi of aged rats,involving synaptic vesicle transport efficiency,synaptic structural and functional plasticity,intracellular protein homeostasis,NMDA-mediated Ca2+signal transport,energy metabolism,and etc.which may be involved in the mechanism of ketamine-induced temporary cognitive dysfunction.

Objective: To evaluate the effects of ketamine anesthesia on proteome in hippocampus in aged rats. Methods: Thirty healthy male Wistar rats, aged 20 months, weighing 560-610 g, were divided into 2 groups (n=15 each) using a random number table method: control group (group C) and ketamine group (group K). In group K, ketamine 80 mg/kg was intraperitoneally injected, additional 1/2 initial dose was given when the righting reflex was recovered, and anesthesia was maintained for 3 h. Morris water maze test was performed starting from 1st day after the end of anesthesia.Five rats were selected at days 1 and 7 after the end of anesthesia and sacrificed, and hippocampal tissues were obtained to extract proteins.Proteins extracted from rat hippocampi were identified by 2-dimensional electrophoresis (2-DE). The differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and biological information system. Results: Compared with group C, the escape latency and total swimming distance to find the submerged platform in Morris water maze at the 1st day after anesthesia were significantly prolonged in group K (P<0.05 or 0.01). The MALDI-TOF-MS analysis showed that there were 21 differentially expressed proteins at 1st day after ketamine anesthesia, of which 6 proteins (involving maintenance of intracellular protein homeostasis, energy metabolism, etc.) presented with up-regulated expression and 15 proteins (involving synaptic vesicle transport efficiency, synaptic structural and functional plasticity, maintenance of intracellular protein homeostasis, NMDA-mediated Ca²⁺ signal transport, energy metabolism, etc.) presented with down-regulated expression.There were 8 differentially expressed proteins at 7th day, including 3 proteins with up-regulated expression and 5 proteins with down-regulated expression (P<0.05). Conclusion Ketamine anesthesia can induce 21 differentially expressed proteins in hippocampi of aged rats, involving synaptic vesicle transport efficiency, synaptic structural and functional plasticity, intracellular protein homeostasis, NMDA-mediated Ca²⁺ signal transport, energy metabolism, and etc.which may be involved in the mechanism of ketamine-induced temporary cognitive dysfunction.