الملخص
Objective To establish a genetic monitoring method for laboratory quails.Methods Quail microsatellite loci were searched in the literature,and microsatellite DNA loci suitable for quails were screened by an interspecific transfer method in closely related species,namely chickens and ducks.Quail liver DNA was extracted as a template,and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis.On the basis of amplification of the selected microsatellite loci,the number of alleles,polymorphisms,and microsatellite loci combinations for quail genetic quality detection were selected and detection method were developed.Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails.Conclusions A genetic monitoring method for laboratory quails was preliminary developed.
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Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×104 U and 33.22×104 U) was higher than that of BALB/c mice (5.33×104 U and 19.31×104 U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×104 U/g) was higher than that of BALB/c mice (0.23×104 U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.
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Objective@#To research the auditory nerve transduction effects under multi-wavelength pulsed laser stimulations within a safe and acceptable signal range.@*Methods@#The real-time detection of intracellular calcium concentration was adopted by specific fluorescent indicator staining based on calcium imager. The spiral ganglion cells of mice were cultured in vitro. After fluorescent indicating, morphologic observation under optical microscope, Fura-2 calcium ion fluorescence excitation, intact morphology cells selection, fixing the optical fiber, the spiral ganglion cells were irradiated by different wavelength laser, including visible light (450 nm) and near infrared light (808 nm,1 065 nm). The intracellular calcium concentration was monitored by calcium ion imaging.@*Results@#When 450 nm laser stimulated spiral ganglion cells, the intracellular calcium concentration was strongly increased, however, for other wavelength laser stimulation, there was no obvious relative response. And the sensitivity expression of the nerve cells under laser was related with the location of laser fiber. Cells closer to the fiber produced more obvious changes in calcium ion concentration, while for cells farther away from the fiber, the change amplitudes were weaker although the number of changes in calcium ion concentration was consistent.@*Conclusion@#The spiral ganglion cells of mice can induce a signal transduction response under the action of laser, and the response has laser wavelength selectivity.
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Objective To investigate the morphological and immunophenotypic characteristics of phyllodes tumors (PT) and explore their significance in differential diagnosis and evaluation of prognosis.Methods 48 specimens with PT were observed under light microscope with HE and immunohistochemical staining (EnVision method).The antibodies including CD34,CD10,CD117,p53,and Ki-67 were used.10 cases of fibroadenoma were compared and the clinical data and follow-up results were analyzed retrospectively.Results All of 48 cases with PT presented as well-circumscribed masses with typically leaflike structures composed of double-layered epithelial component arranged in clefts and overgrowing hypercellular mesenchymal component.31 benign,12 borderline,and 5 malignant PT were diagnosed based on stromal overgrowth,cellular pleomorphism,mitosis,margins,and others.Focal necrosis was detected in 3 malignant cases and liposarcoma component existed in 1 case.Immunohistochemical staining showed that the positive rates of CD34 were 93.5 ％,58.3 ％,0 in benign,bordline,and malignant PT respectively,there were significant differences between each groups (P ＜ 0.05).The expression of CD10 were 25.8 ％,83.3 ％,80.0 ％,its expression in benign group was significantly different compared with bordline and malignant PT (P ＜ 0.05).p53 was expressed in three groups with no significant difference (P ＞ 0.05).CD117 and Ki-67 had high expression comparatively in malignant PT.All cases were treated surgically with a local recurrence rate of 22.9 ％.Conclusions Reasonable surgery pattern based on accurate histopathological diagnosis is crucial to reduce the local recurrence rate of PT.Immunohistochemistry examination in PT is helpful to the differential diagnosis and evaluation of prognosis.
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Serum B7-H4 protein levels were measured by enzyme-link immunosorbent assay (ELISA) in 62 patients with epithelial ovarian cancer, 20 patients with benign ovarian diseases, and 30healthy subjects.Serum B7-H4 protein levels in epithelial ovarian cancer, benign ovarian diseases and normal controls were 0.72(0.32,1.88 )μg/L, (0.45±0.21)μg/L,(0.38 ± 0.19 ) μg/L respectively (P＜0.01).The serum level of B7-H4 protein in epithelial ovarian carcinoma was correlated with FIGO stages, lymph node metastasis, histological grade and histological types ( all P ＜ 0.01 ), but not with menopause and ascites (P＞0.05).The receiver operating characteristics curve analysis revealed that B7-H4protein had a sensitivity and specificity of 69% (43/62)and 86%(43/50) for diagnosis of epithelial ovarian cancer.
الملخص
Objective To investigate the change and clinical significance of the serum level of human epididymis secretory protein 4 (HE4) in elderly patients with epithelial ovarian cancer (EOC).Methods The serum levels of HE4 were determined by ELISA in EOC patients (n= 33), benign ovarian disease patients ( n = 17) and healthy women ( n = 20), all of them aged 60 years or over. And there were other 31 EOC patients and 20 healthy women aged less than 60 years at the same period as controls. Results In healthy people, the serum level of HE4 was higher in the group with age ≥60 years than with age ＜60 years [(32.25± 13. 15) pmol/L vs. (24.59±8.60) pmol/L]. The serum HE4 level had a positive correlation with age (r=0. 525, P＜0.01). The median level of HE4 was significantly higher in elderly EOC patients than in benign ovarian disease patients (81.50 pmol/L vs.45.60 pmol L, U=168.5, P＜0.05) and healthy women (81.50 pmol L vs. 33.30 pmol L, U=76.5, P＜0. 01). The HE4 had a significantly higher area under ROC curve when compared with CA125 (0. 799 vs. 0. 782), while the cut-off value was set 65.43 pmol L with a sensibility and specificity of 66.67% and 86.49% respectively. The diagnostic efficacy of HE4 was higher than CA125. There was no significantly statistical difference between elderly and non-elderly EOC patients in serum HE4 levels (U= 101.2, P＞0. 05). Conclusions Age is an important factor which can affect human serum level of HE4. HE4 determination is helpful in the diagnosis of ovarian epithelial carcinoma in the elderly patients, and the combined detection of HE4 and CA125 is an useful adjuvant diagnostic measure.