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Objective To prepare a rabbit anti-mouse coiled-coil domain containing 189 (Ccdc189) polyclonal antibody. Methods The pET-28a-Ccdc189 prokaryotic expression plasmid was constructed and transformed into E.coli BL21. IPTG was used to induce the expression of Ccdc189 prokaryotic protein. Adult male New Zealand rabbits were immunized with purified recombinant protein to obtain rabbit anti-mouse Ccdc189 polyclonal antibody. The specificity of the polyclonal antibody was identified by Western blot analysis, indirect ELISA and immunofluorescence histochemical staining. Results The pET-28a-Ccdc189 recombinant plasmid was successfully constructed and the expression of the Ccdc189 recombinant protein was induced. ELISA revealed that the titer of the polyclonal antibody was 1:1 000 000. Western blot and immunofluorescence staining demonstrated that the Ccdc189 polyclonal antibody could specifically identify the Ccdc189 prokaryotic protein and the Ccdc189 protein in adult wild-type mouse testis. Conclusion A polyclonal antibody with high specificity against mouse Ccdc189 was successfully created.
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Conejos , Masculino , Animales , Ratones , Especificidad de Anticuerpos , Anticuerpos , Ensayo de Inmunoadsorción Enzimática , Western Blotting , Proteínas Recombinantes , Escherichia coli/genéticaRESUMEN
Objective To prepare rabbit polyclonal antibody against mouse IQ and ubiquitin-like domain-containing protein (IQUB) and detect its expression in the mouse testis. Methods Full-length coding sequence of IQUB was inserted into the pET-30a(+) vector to construct pET-30a-IQUB recombinant prokaryotic plasmid. Transformation of pET-30a-IQUB plasmid into E. coli BL21 was performed, and protein expression was induced with isopropyl-beta-D-thiogalactoside (IPTG). The protein was purified through histidine-tagged fusion protein purification column, then denatured by treatment of urea with gradient concentration. New Zealand rabbits were immunized with the denatured protein to produce IQUB polyclonal antibody. Antibody titer was detected by ELISA, and Western blot analysis and immunofluorescence assay were employed to validate the effectiveness and specificity of IQUB antibody. Results pET-30a-IQUB recombinant plasmid was constructed, and protein expression of IQUB was induced successfully with IPTG. The titer of IQUB polyclonal antibody reached 1:1 000 000. The antibody specifically recognized the endogenous IQUB protein of testis in the wild-type adult mouse. IQUB was expressed in spermatogenic cells of different stages. It was localized in the acrosome and flagellum of mature sperms. Conclusion The highly specific rabbit anti-mouse IQUB polyclonal antibody is successfully prepared, which can be used for Western blot and immunofluorescence histochemistry.
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Masculino , Conejos , Animales , Ratones , Ubiquitinas , Escherichia coli/genética , Isopropil Tiogalactósido , Anticuerpos , Ensayo de Inmunoadsorción EnzimáticaRESUMEN
It is shown that phytochemicals have a protective effect on colon cancer. Curcumin, polysaccharides (apple polysaccharides, mushroom glucans), saponins (paridis saponins, ginsenosides), resveratrol, quercetin and other plant drugs can inhibit colon cancer cell proliferation and promote cell apoptosis through different signaling pathways. In addition, it also has anti-inflammatory, antioxidant, anti-angiogenesis, reduce the toxic side effects of chemotherapy drugs, and reverse the drug resistance of tumor cells. Understan-ding the prevention and cure effect of plant medicine on colon cancer and its possible mechanism can provide more theoretical basis and therapeutic ideas for the clinical prevention and cure of colon cancer.
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Objective To explore the related factors of senile acute pulmonary thromboembolism(PTE) patients,and to observe the prognosis and analyze the strategy of control and prevention to reduce the incidence. Methods 50 elderly patients with acute PTE were selected,related factors were analyzed by using the retrospective investigation,and the prevention and control were administrated.Results The mainly disease occurred of the elderly patients with acute PTE were combined with chronic diseases 42% (21 /50),with lower extremity DVT 26.00%(13 /50)and with recent trauma,fracture and operation 20%(10 /50).Elderly patients with acute PTE who were ≥70 years of age and lying in bed the time≥1 month,were 58%(29 /50)and 62%(31 /50),which were higher than those of the other age <70 years old and lying in bed time <1 month of patients,the difference was statistically signifi-cant(χ2 =9.26,9.80,all P <0.05).Conclusion The related risk factors for elderly patients with acute PTE mainly correlate with age,lying in bed time,chronic diseases,patients with lower extremity DVT and recent trauma,fracture, operation,the variety of factors and overlap each other.Mastering the variety factors of PTE in the nursing process,and taking effective measures to prevent and control the factorscan reduce or avoid the occurrence of acute PTE,and im-prove the prognosis of the patients.
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The liver is the central organ involved in lipid metabolism. Dyslipidemia and its related disorders, including non-alcoholic fatty liver disease (NAFLD), obesity and other metabolic diseases, are of increasing public health concern due to their increasing prevalence in the population. Besides their well-characterized functions in cholesterol homoeostasis and nutrient absorption, bile acids are also important metabolic regulators and function as signaling hormones by activating specific nuclear receptors, G-protein coupled receptors, and multiple signaling pathways. Recent studies identified a new signaling pathway by which conjugated bile acids (CBA) activate the extracellular regulated protein kinases (ERK1/2) and protein kinase B (AKT) signaling pathway via sphingosine-1-phosphate receptor 2 (S1PR2). CBA-induced activation of S1PR2 is a key regulator of sphingosine kinase 2 (SphK2) and hepatic gene expression. This review focuses on recent findings related to the role of bile acids/S1PR2-mediated signaling pathways in regulating hepatic lipid metabolism.
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OBJECTIVE To discuss the function of sustaining quality improvement in preventing perineum wound infection.METHODS The perineum wound infection situation in 2003 was retrospectively analyzed,and since then the measures for sustaining quality improvement were implemented.RESULTS In 2004 and in 2005 the perineum wound infection percentage and the stitch inflammation rate were obviously reduced than in 2003.The difference was statisticaly significant.CONCLUSIONS Sustaining quality improvement is an unceasing,gradually advancing and long-term process.The prevention of perineum wound infection is a long time work for obstetrics workers.To carry out strictly the aseptic technical operation,grasp skilled suture technology,choose easily absorbable suture,maintain suitable cleanliness and nursing and use the antibiotic could be effectively to prevent the perineum wound infection and the stitch inflammation occurrence.