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This study used the zebrafish model to explore the hepatotoxicity of Rhododendri Mollis Flos(RMF). The mortality was calculated according to the number of the survival of zebrafish larvae 4 days after fertilization under different concentration of RMF, and the dose-toxicity curve was fitted to preliminarily evaluate the toxicity of RMF. The liver phenotypes under the sublethal concentration of RMF in the treatment group and the blank control group were observed by hematoxylin-eosin(HE) staining and acridine orange(AO) staining. Meanwhile, the activities of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were determined to confirm the hepatotoxicity of RMF. Real-time quantitative polymerase chain reaction(real-time PCR) and Western blot were used to determine the expressions of genes and proteins in zebrafish larvae. Gas chromatography time-of-flight mass spectrometry(GC-TOF-MS) was used to conduct untargeted metabolomics testing to explore the mechanism. The results showed that the toxicity of RMF to zebrafish larvae was dose-dependent, with 1 100 μg·mL~(-1) of the absolute lethal concentration and 448 μg·mL~(-1) of sublethal concentration. The hepatocyte apoptosis and degeneration appeared in the zebrafish larvae under the sublethal concentration of RMF. The content of ALT and AST in zebrafish larvae at the end of the experiment was significantly increased in a dose-dependent manner. Under the sublethal concentration, the expressions of genes and proteins related to apoptosis in zebrafish larvae were significantly increased as compared with the blank control group. The results of untargeted metabolomics showed that the important metabolites related to the he-patotoxicity of RMF were mainly enriched in alanine, aspartic acid, glutamic acid, and other pathways. In conclusion, it is inferred that RMF has certain hepatotoxicity to zebrafish larvae, and its mechanism may be related to apoptosis.
Subject(s)
Animals , Zebrafish/genetics , Apoptosis , Larva , Chemical and Drug Induced Liver InjuryABSTRACT
Objective To investigate the evaluation of the doctor-patient trust by family members based on the general interpersonal trust scale, and explore the influencing factors of the evaluation of doctor-patient trust by patient family members, and to provide references for improving the doctor-patient trust and the doctor-patient relationship. Methods A survey of 3010 patient family members in 10 hospitals from 5 provinces in China was conducted using a self-made questionnaire and a general interpersonal trust scale. Results Only 47.2% of the family members agreed that the doctor-patient trust degree was good, and the interpersonal trust score was (70.52±7.07) points. The age, monthly income level, life satisfaction of family members are the important factors affecting the evaluation of doctor-patient trust (P < 0.05). Conclusion The patient family members have low recognition of doctor-patient trust, and some targeted measures should be taken to improve the interpersonal trust level and life satisfaction of the family members, so as to further enhance the doctor-patient trust.
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OBJECTIVE: To observe the effect of "Neiguan" (PC6)-electroacupuncture (EA) preconditioning on serum metabolites in myocardial ischemia-reperfusion injury (MIRI) rats, so as to reveal its mechanism underlying improvement of ischemic myocardium from metabonomics. METHODS: A total of 48 male SD rats were randomly divided into control, model, EA "Neiguan"(PC6) and EA "Hegu"(LI4) groups (n＝12 rats/ group). Rats of the control group were just banded on animal boards for 30 min, once daily for 7 days. The MIRI model was established by occlusion of the left anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 1 h, and rats of the model group were also banded as those in the control group. Before modeling, EA (10 Hz/50 Hz, 1 mA) was applied to bilateral "Neiguan"(PC6) and "Hegu"(LI4) for 30 min, once daily for 7 successive days. After the treatment, serum samples were collected to be analyzed by proton nuclear magnetic resonance (1H NMR) spectroscopy. The orthogonal partial least squares discriminate analysis (PLS-DA) was employed to distinguish the serum differential metabolic profile of rats in different groups and identify potential biomarkers. RESULTS: After modeling, the ECG of model group and electroacupuncture groups showed T wave towering, and there was no obvious ST segment between R wave and T wave. The T wave decreased more than 0.2 mV after reperfusion, and there was no obvious ST segment. Compared with the control group, MIRI induced significant changes of metabolites in the serum including increase of acetoacetate acid, lectic acid, creatine, glycerol and glucose, and decrease of alanine, glutamine, glycerophosphoryl choline and phosphorylcholine. In comparison with the model group, PC6-EA preconditioning induced significant changes, including an increase of glucose, and a decrease of leucine，isoleucine, valine，3-hydroxybutyric acid，lactate，acetate，acetone，acetoacetate acid，pyruvic acid，glutamine，creatine and glycerol. There is no significant difference in metabolic patterns between "Hegu" group and model group. Metabolic pathway enrichment analysis indicated that the protective effect of PC6-EA pretreatment was realized mainly by regulating pathways of glycolysis, gluconeogenesis, citric acid metabolism, pyruvate metabolism, ketone body metabolism, etc. CONCLUSION: PC6-EA pretreatment has a role in regulating gluconeogenesis, pyruvate metabolism, amino metabolism, ketone body metabolism and energy metabolism in rats with MIRI, which maybe contribute to its protective effect on ischemic myocardium, but the specific metabolic pathways and mechanisms need being studied further.
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Objective:To observe the effect of electroacupuncture (EA) pretreatment on adenine nucleotides in the myocardial tissues of the myocardial ischemia-reperfusion injury (MIRI) rats, and to explore the mechanism of EA pretreatment on myocardial prevention and protection in MIRI rats. Methods:Forty SPF male Sprague-Dawley (SD) rats were randomly divided into 5 groups: a blank group, a sham operation group, a model group, an EA at Neiguan (PC 6) group and an EA at Hegu (LI 4) group, with 8 rats in each group. Rats in the blank group only received binding to the rat plate, 30 min/time, once a day for 7 d; on the 7th day, rats in the sham operation group were subjected to threading for 40 min at the left anterior descending coronary artery without ligation, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day, rats in the model group were subjected to threading at the left anterior descending coronary artery with ligation, for 40 min before the blood flow was restored, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day of pretreatment with EA at Neiguan (PC 6) or Hegu (LI 4) for 30 min per day (once a day for 7 d), rats in the EA at Neiguan (PC 6) group and EA at Hegu (LI 4) group were subjected to modeling and sample collection same as in the model group. The left ventricular myocardium of the lower left anterior descending coronary artery was collected from rats in all 5 groups. Hematoxylin-eosin (HE) staining and transmission electron microscope (TEM) were used to observe the changes in myocardial pathological morphology. The change in the adenine nucleotide level of myocardial tissue was measured by high performance liquid chromatography (HPLC). Results:The HE staining and ultrastructure showed that the myocardial injury was severer in the model group compared with the sham operation group. Compared with the model group, the myocardial injury in the EA at Neiguan (PC 6) and the EA at Hegu (LI 4) groups was mild or hardly any. The adenine nucleotide levels in the sham operation group and the model group were all decreased compared with the blank group (allP<0.05); compared with the sham operation group, the adenine nucleotide level of the model group was also decreased, but the difference was not statistically significant (P>0.05); compared with the model group, the adenine nucleotide level in the EA at Neiguan (PC 6) group was increased (P<0.05), and the adenine nucleotide level in the EA at Hegu (LI 4) group was significantly increased (P<0.01). The adenine nucleotide level in the EA at Hegu (LI 4) group was higher than that in the EA at Neiguan (PC 6) group, but the difference was not statistically significant (P>0.05). Compared with the EA at Neiguan (PC 6) group, the levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in the EA at Hegu (LI 4) group were significantly increased (allP<0.01). Conclusion:Both EA at Neiguan (PC 6) and Hegu (LI 4) can alleviate the pathological damage to myocardium in MIRI rats, and increase the adenine nucleotide level in myocardial tissues, and thus protect MIRI rats. EA at Hegu (LI 4) has a better protective effect than Neiguan (PC 6).
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<p><b>OBJECTIVE</b>To report an X-linked dominant Charcot-Marie-Tooth disease (CMTX) Chinese family with vocal cord paresis and to identify the mutation of gap junction protein beta 1 gene (GJB1).</p><p><b>METHODS</b>Part of the family members with dysphagia, dysphonia and lethal respiratory failure were studied through flexible laryngoscope, clinical, brain MRI and electrophysiological examinations. After excluding large fragment tandem duplication containing peripheral myelin protein 22 gene (PMP22), direct sequencing was performed to analyze the mutation of the GJB1 gene in 5 patients including the proband, 5 unaffected family members and 50 unrelated healthy individuals.</p><p><b>RESULTS</b>Eight members spanning 3 generations in this family were affected with CMTX characterized by progressive atrophy and weakness of the anterior tibial and peroneal muscles, especially in the proband. Vocal cord paresis was observed through flexible laryngoscope in total of 4 affected members with dysarthria and dysphagia, 2 of them died of severe respiratory failure due to complete bilateral vocal cord involvement. Normal brain MRI was observed in the proband. The electrophysiological data showed predominant demyelization involving the motor and sensory nerves in the proband. DNA sequencing revealed a de novo c.186 C>G missense mutation in exon 2 of the GJB1 gene, the mutation cosegregated with phenotype.</p><p><b>CONCLUSION</b>Respiratory failure associated with vocal cord involvement may be a rare and severe symptom in CMTX. The present report provides further evidence for clinical and genetic heterogeneity in the X-linked Charcot-Marie-Tooth disease.</p>