ABSTRACT
Background Research on non-target organ damage of biological pesticides has attracted much attention. Rotenone exposure may be far beyond the occupational environment, and the exposureduring pregnancy may be increased through bioaccumulation, fruit or vegetable residues, and other forms of oral intake. At present, the effects of rotenone on placental development and its mechanism are still unknown. Objective To investigate the developmental damage of rat placenta and evaluate the expression levels of glycogen synthase kinase (GSK-3β) and beta catenin (β-catenin) followed by rotenone exposure through the placental barrier during pregnancy, as well as to propose possible associated mechanisms. Methods Eighteen sexually mature SD female infertile rats without specific pathogens were selected and divided into three groups: blank control group (0.9% saline), corn oil group, and rotenone group (corn oil + 2 mg·kg−1 rotenone) by random number method, six female animals in each group. Another six male rats were selected and mated to the female rats at night with a female to male ratio of 3:1 per cage. Pregnant rats were given 0.9% saline, corn oil, and 2 mg·kg−1 rotenone preparation by isovolumetric gavage once daily for the entire gestation period (19 d), and their conditions were observed after the last dose. The pregnant rats were anesthetized, and the size of the placenta and blood perfusion were detected by ultrasound the next day of the last dose of rotenone. Then, 3 pregnant rats in each group were sacrificed immediately and the placenta and umbilical cord tissues were dissected. The remaining 9 pregnant rats gave birth naturally, and the fetuses were observed for developmental evaluation and weighed. The histopathological changes of umbilical cord and placenta were observed by hematoxylin-eosin staining. The reactive oxygen species levels of placenta tissues were detected by flow cytometry. The Ca2+-ATPase activity of placenta tissues was detected by colorimetric method. The localization and levels of GSK-3β and β-catenin expression of placenta were detected by immunohistochemistry. The p-GSK-3β/GSK-3β and p-β-catenin/β-catenin protein expression in placental tissues were measured by Western blotting. Results No injury or death was recorded during the pregnant rats receiving rotennon administration. Adverse pregnancy outcomes such as fetal absorption and postpartum stillbirth were found in the rotenone group, and the weight of the fetal mice decreased (P<0.05). The B-ultrasound showed disc-shaped placenta with a thick middle and thin edge, smooth fetal surface, rough maternal surface, visible placental lobules, granular echotexture of the placenta with comma-like echogenic densities, and chorionic plate showing deep indentations, no calcification, degeneration, or necrosis in each group. Compared with the corn oil group, the fetal surface diameter of the placenta was reduced in the rotenone group (P<0.05). The Doppler color ultrasound showed that interplacental blood flow was reduced in the rotenone group, while interplacental blood flow was abundant in the blank control and the corn oil groups. The hematoxylin-eosin staining results showed that smooth muscle cells in the umbilical cord tissues of rats were loosely arranged, with fuzzy nuclei and inflammatory infiltration in the rotenone group. The placental trophoblast cells were small in size, disorderly arranged with nuclear fragmentation and cytoplasm turbidity. The tissue reactive oxygen species level in the rotenone group was higher than those in the other two groups (P<0.05). The Ca2+-ATPase activity of placental tissues was reduced in the rotenone group (P<0.05). The immunofluorescence low-magnification observation showed that GSK-3β and β-catenin were expressed in placental tissue, weak fluorescence expression in the decidua basalis, strong fluorescence expression in the labyrinthine layer structure. The labyrinthine layer under high magnification showed that compared with the blank control group and the corn oil group, the brightness of β-catenin fluorescence expression in the rotenone group decreased (P<0.05), and the brightness of GSK-3β expression increased (P<0.05). The Western blotting results showed that the expression of β-catenin and p-GSK-3β proteins decreased (P<0.01), and the expression of GSK-3β protein increased (P<0.01) in the rotenone group. No significant expression of p-β-catenin protein was detected in the placenta tissue of each group. Conclusion Rotenone exposure during pregnancy induces placental hypoperfusion, growth retardation, and oxidative stress in rats, as well as down-regulation of β-catenin and p-GSK-3β protein expression, and up-regulation of GSK-3β protein expression, which may further lead to abnormal pregnancy and fetal restricted growth.
ABSTRACT
OBJECTIVE： To provide reference for strengthening clinical application of key monitoring drugs and promoting rational drug use in clinic. METHODS： Based on evidence-based medicine， taking key monitoring drugs Shuxuetong injection as example， clinical evidence of domestic and foreign clinical studies were collected. The included literatures were graded according to the quality of GRADE evidence and recommended strength system. Evidence-based medicine evidence for the indications of Shuxuetong injection were evaluated， and criterion for clinical use of Shuxuetong injection was formulated in Huaihua First People’s Hospital （our hospital）. RESULTS： The main content of criterion for clinical application of Shuxuetong injection formulated by our hospital was that there was A-level evidence support for acute ischemic cerebral infarction， but it was weakly recommended and only used for adjuvant therapy； there was B-level evidence support for anticoagulation （for preventing DVT）， diabetic peripheral nerve lesion， but it was weakly recommended； there was only C-level or D-level evidence support for other indications， it was strongly recommendation against use. CONCLUSIONS： Clinical pharmacists formulate the criterion for clinical application of Shuxuetong injection by evidence quality evaluation method， provide reference for clinical application management of key monitoring drug and play an important effect on rational drug use in clinic.
ABSTRACT
Objective To explore the value of coronary flow reserve(CFR) evaluating myocardial ischemia measured by adenosine triphosphate (ATP) stress transthoracic Doppler echocardiography (TTDE),and the feasibility of CFR to predict coronary stenosis.Methods Fifty-four patients suffering chest pain with known or suspected coronary artery disease were performed ATP stress TTDE to measure resting and maximum expansion coronary blood flow velocity and calculate CFR.all patients were performed by coronary angiography (CAG) and single-photon emission computed tomography (SPECT) myocardial perfusion imaging.Results ① To evaluate myocardial ischemia,there was not statistical significant difference between non-invasive CFR and SPECT myocardial perfusion imaging(P ＞0.05).CFR≤2.0 was the best cutoff value for evaluating myocardial ischemia which yielded a sensitivity of 93.3 ％ and specificity of 89.7％.②Coronary artery stenosis was negatively correlated with CFR (P ＜0.001).ROC curve analysis demonstrated that CFR≤ 1.60 yielded a sensitivity of 92.3％ and specificity of 73.3％ to predict coronary stenosis significantly.Conclusions CFR measured by ATP stress TTDE can evaluate myocardial ischemia of coronary artery disease and predict LAD significant stenosis before CAG.Using CFR and CAG has important clinical value for choosing treatment of stable coronary artery disease.