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Objective@#To study the effects of Lycium barbarum polysaccharides ( LBP ) on blood indexes and liver tissue morphology in rats with intrahepatic cholestasis.@*Methods@#Sprague-Dawley rats were randomly divided into the control group, the model group, and LBP low, medium and high dose group. The rats in the model group and LBP dose groups were given 60 mg/kg alpha-naphthylisothiocyanate ( ANIT ) by gavage every three days of the experiment, and the rats in the control group were given salad oil instead of ANIT. From the third day, the rats in each dose group were given 40, 150 and 600 mg/kg LBP, and the rats in the model group were given distilled water. After four weeks, the blood and urine indexes were measured, and the morphological changes of liver tissue were observed. @*Results@#From the third day of the experiment, the activity of rats in the model group and LBP dose groups decreased, and the color of urine changed to dark yellow. There was no abnormality in the group. In the model group, the levels of serum total bilirubin, direct bilirubin, total bile acid ( TBA ), alkaline phosphatase ( ALP ), γ-glutamyltransferase(γ-GGT), cholesterol, alanine aminotransferase ( ALT ), aspartate aminotransferase ( AST ), white blood cell ( WBC ), percentage of granulocyte, urinary bilirubin, urinary bile acid, liver mass and liver to body ratio were higher than those in the control group, while red blood cell and percentage of lymphocyte were lower than those in the control group ( all P<0.05 ). Pathological changes of liver tissue were observed. The levels of serum TBA, ALP, γ-GGT, ALT, AST, WBC and liver to body ratio in LBP high dose group were lower than those in the model group ( all P<0.05 ). The infiltration of inflammatory cells, proliferation and expansion of bile duct, degeneration and necrosis of liver cells were alleviated. @*Conclusions@#LBP can improve the blood indexes and pathological changes of liver tissue in rats with intrahepatic cholestasis at the dosage of 600 mg/kg. Inhibition of inflammatory response and reduction of oxidative stress injury may be the mechanism for alleviating cholestatic liver injury.
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SUMMARY: Fixation is a crucial step in processing of tissue specimen for preservation of cellular architecture and composition of cells. Alcohol-based fixatives are considered some of the most promising alternatives to formalin. We evaluated the performance of alcohol-based fixatives (EthMeth and methacarn) and formalin as a comparator fixative in the research laboratory. Following 24 hours of fixation, tissue morphology and cellular details of the liver, spleen and brain (cerebral cortex) were evaluated. Morphological characteristics were evaluated by gross observations and analyzing cellular details, tissue architecture and overall staining characteristics (Hematoxylin and Eosin). EthMeth and methacarn fixation gave generally comparable and satisfactory results on the tissue morphology and subsequent identification of tissue characteristics. Particularly, tissues were well preserved and all nuclear as well as cytoplasmic details were clearly visible. However, formalin fixed tissues showed some peculiarity such as improper fixation, mild shrinkage, and alterations of tissue components. These results confirm that alcohol-based fixation is the superior alternative to formalin for preservation of tissue morphology. However, it is required to standardize the formalin-free methods and harmonize diagnosis in the laboratory worldwide.
RESUMEN: La fijación es un paso crucial en el procesamiento de muestras de tejido para preservar la arquitectura celular y la composición de las células. Los fijadores a base de alcohol se consideran algunas de las alternativas más prometedoras a la formalina. Evaluamos el rendimiento de los fijadores a base de alcohol (EthMeth y metacarn) y formalina como fijador comparativo en el laboratorio de investigación. Después de 24 horas de fijación, se observó la morfología del tejido y los detalles celulares del hígado, bazo y corteza cerebral. Se evaluaron las características morfológicas mediante observaciones generales y analizando detalles celulares, arquitectura de tejidos y características generales de tinción (hematoxilina y eosina). La fijación de EthMeth y metacarn dio resultados generalmente comparables y satisfactorios en la morfología del tejido y la posterior identificación de las características del mismo. Particularmente, los tejidos estaban bien conservados y todos los detalles nucleares y citoplasmáticos eran claramente visibles. Sin embargo, los tejidos fijados con formalina mostraron cierta peculiaridad, tal como una fijación inadecuada, la contracción leve y alteraciones de los componentes del tejido. Estos resultados confirman que la fijación a base de alcohol es la mejor alternativa a la formalina, para preservar la morfología del tejido. Sin embargo, es necesario estandarizar los métodos sin formalina y armonizar el diagnóstico en los laboratorios.
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Fijación del Tejido/métodos , Alcoholes/química , Fijadores , Formaldehído/química , Cloroformo/química , Ácido Acético/química , Metanol/químicaRESUMEN
Objective: The exploration of the anticancer potential of polysaccharide isolated from the methanolic extract of Tinospora cordifolia (T. cordifolia) stem bark against breast cancer in DMBA-induced female albino Wistar rat models were examined by various hematological parameters. Methods: Analysis of Red blood cell (RBC), White blood cell (WBC) and platelet level, Tumor markers Carcino Embryonic Antigen (CEA) and Cancer Antigen 15.3 (CA 15.3) in the serum, was done in the normal, cancer and compound treated rats using specific kits. Histological studies were performed to examine the changes in the tissue morphology and cell patterns in breast tissue. Results: The decreased levels of RBC, WBC and platelets in 7,12-Dimethylbenz [a] anthracene (DMBA)-induced breast cancer (Group III) animals were revived to the normal conditions in polysaccharide treated breast cancer (Group IV) animals as that of normal (Group I). The level of tumor markers CEA and CA 15.3, was found elevated in serum of DMBA-induced breast cancer groups (Group III) when compared to their levels in the normal groups (Group I) whereas polysaccharide treatment (Group IV) prevented this rise in the levels of tumor markers. The histological studies on the breast tissue samples of all the groups showed the appropriate features where the normal (Group I) animals were characterized with normal cells uniformly arranged without any change in orientation and morphology, DMBA-induced cancer (Group III) animals showed an improper orientation of cells arranged as glandular structures, as nest, or cords of various sizes or as solid sheets foci of necrosis in some areas with margins infiltrating, pushing, circumcised or mixed and the polysaccharide treated (Group IV) animals showed results resembling that of the normal (Group I) animals. Conclusion: Thus, polysaccharide is proved as an effective chemo preventive agent against breast cancer.
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Objective: To investigate the significance of speckle tracing echocardiography in the examination of the rat models of myocardial infarction, and to elucidate the reliability and clinical application of speckle tracing echocardiography in the identification of the myocardial infarction rats. Methods: A total of 50 rats were randonly divided into sham operation group and model group, with 25 rats in each group. Ischemic preconditioning method was used to establish the myocardial infarction models. Speckle tracking echocardiography and cardiac tissue Masson staining were performed to analyze the changes of echocardiography and the morphology of myocardium tissue of the rats. Results: Compared with sham operation group, the each index in echocardiography of the rats in model group before modeling had no significant differences ( Pr>0. 05). Compared with sham operation group∗ the left ventricular ejection fraction (KF), circumferential strain (CS)» the peak values of CS in the anterior septum, anterior wall, anterior lateral wall, systolic period, radial strain ( RS) of the anterior wall of the rats in model group were decreased ( P<0. 05); the standardized time to peak of CS of anterior wall (TTP) was increased ( P<0. 05). The results of Masson staining showed that there was no change in the myocardium tissue and no infarction of the rats in control group; the infarction area of the rats in model group was (4. 52± 1. 41) %•, the infarction located in the anterior wall which was the myocardiocyte lesion in the small area. Conclusion: Speckle tracing echocardiography RS and IIP can reveal the infarction area and its surrounding areas of myocardial infarction, and quantitatively detect the myocardial infarction site in the rats. Speckle tracing echocardiography can be used for the clinical rapid diagnosis of myocardial infarction and confirmation of infarction area.
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Objective: To investigate gastrointestinal regulation of red yeast rice on spleen deficient dyspepsia mice, and provide reference for clarifying mechanism of invigorating spleen to promote digestion of fermented traditional Chinese medicine. Method: Mice were randomly divided into 5 groups, including the blank group, the model group, the unfermented red yeast rice treatment group (1.17 g·kg-1·d-1), the fermented red yeast rice treatment group (1.17 g·kg-1·d-1), the domperidone group (3.9 mg·kg-1·d-1), ten mice in each group were continuous intragastric administration for 10 days. The effects of red yeast rice before and after fermentation on the general behavior, body weight, food intake and gastrointestinal function of spleen deficient dyspepsia mice were compared. The effects of red yeast rice on the gastrointestinal hormone[motilin (MTL), gastrin (GAS), 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP)] levels in serum of spleen deficient dyspepsia mice were determined by enzyme-linked immunosorption assay (ELISA). The regulation function of red yeast rice on the pathological changes of gastrointestinal tissue of spleen deficient dyspepsia mice was investigated by hematoxylin-eosin (HE) staining. The effect of red yeast rice on gut microbiota of spleen deficient dyspepsia mice was investigated, the primers were designed on bacterial 16S rRNA V3-V4 region sequences and Illumina Miseq platform was used for high-throughput sequencing. Result: Compared with the model group, fermented red yeast rice could recover the body weight and food intake, reduce gastric residual rate (PPEscherichia-Shigella, while increasing the relative abundance of the beneficial bacteria such as Lactobacillus and Bifidobacterium. Conclusion: Fermented red yeast rice can enhance the gastrointestinal function of spleen deficient dyspepsia mice, the mechanism may be related to regulating gastrointestinal hormone level, improving histopathology of gastrointestinal tissue and restoring intestinal flora structure.
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OBJECTIVE: To observe the effect of acupuncture combined with medication (Rule Granule) on serum prolactin (PRL)，estradiol (E2) and progestone (P) contents and expressions of prolactin receptor (PRLR), estrogen receptor (ER) and progestrone receptor (PR) proteins in mammary gland (MG) tissues of MG hyperplasia (MGH) rats, so as to explore its mechanisms underlying improvement of MGH. METHODS: A total of 55 female SD rats were randomized into blank control group (n＝10), model group (n＝10), acupuncture group (n＝11), medication group (n＝10), and acupuncture plus medication group (n＝9). The MGH model was established by muscular injection of E2 benzoate(0.5 mg/kg, once daily for 20 days), followed by injection of P (5 mg/kg) into the lateral muscle of the hind-limb, once daily for 5 days, and the rats of the control group were treated by muscular injection of normal saline at the same site and in the same procedures. Acupoint group A composed of bilateral Tianzhong (SI 11), Ganshu (BL 18) and Zusanli (ST 36), and group B composed of Tanzhong (CV 17), and bilateral Wuyi (ST 15) and Hegu (LI 4) were alternatively punctured with filiform needles and stimulated swiftly by twirling the needle in each acupoint for 20－30 sec, once daily for 30 days. Rats of the medication group and acupuncture ＋medication group were treated by gavage of Rule Granule fluid containing Baishao (Paeonia Iactiflora Pall), Danggui (Angelica sinensis), etc. (1.5 mL/100 g for each rat) and those of the other groups treated by gavage of distilled water (1.5 mL/100 g). The treatment was given once daily for continuous 30 d. At the end of the experiments, the rats' abdominal aorta blood was collected for assaying the contents of serum PRL, E2 and P with ELISA, and the pathological changes of breast tissue (the left 2nd pairs) were observed under microscope after sectioning and H．E. staining. The expression of PRLR, ER and PR proteins in the breast tissue was detected by immunohistochemistry. RESULTS: After modeling, the hyperplasia of mammary gland in the model group was obvious; the serum PRL and E2 contents and the immunoactivities of PRLR, ER and PR were significantly increased (P0.05)．. CONCLUSION: Acupuncture, Rule Granule, and acupuncture combined with Rule Granule can improve hyperplasia of mammary gland in rats, which may be related to their effects in reducing serum PRL and E2 and breast ER, PR and PRLR expression levels, and in increasing serum P level. The therapeutic effects of acupuncture plus Rule Granule are better than those of simple acupuncture and simple Rule Granule.
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OBJECTIVE:To observe the effects of aerosol inhalation of Chuankezhi injection on lung index and tissue morphol-ogy of trachea,lung and esophagus in guinea pigs. METHODS:40 guinea pigs were randomly divided into blank control group (normal saline),Chuankezhi high-dose,medium-dose and low dose (2.48,1.24 and 0.62 ml/kg) groups. Each treatment group was administrated by inhaling the atomized injection,once a day,20 ml/kg,for consecutive 2 weeks. The general situation of guin-ea pigs was observed 1 h after each time of medication;1 h after last medication,lung index of guinea pigs was detected,as well as the tissue morphology of left lung,right lung,trachea and esophagus by HE staining. RESULTS:Compared with normal control group,the general situation of guinea pigs were normal in Chuankezhi high-dose,medium-dose and low dose groups;there was no difference in lung index and all of them were about 0.653;tissue section staining showed that no abnormal lesion was found in these tissues. CONCLUSIONS:2 weeks of aerosol inhalation of Chuankezhi injection have no significant effect on lung index and the tissue morphology of trachea,lung and esophagus of guinea pigs.