The pharmaceutical ethynylestradiol (EE) is a potent endocrine modulator. Application enlargement of ethynylestradiol in clinics and abuse in livestock farming and fishing make it important to explore ethynylestradiol toxicological action on vertebrate embryonic development and to establish an in vivo method for EE toxicity detection efficiently and conveniently. In the present study, using a model animal zebrafish and 17alpha-ethynylestradiol as a representative compound, we have investigated EE2 teratogenicity, target tissues and target genes on zebrafish embryo. The results show that median teratogenesis concentration (TC50) of EE2 is 0.8 microg x mL(-1), and median lethal dose (LD50) is 3.3 microg x mL(-1). Targets of EE2 action were implicated in brain, eyes, heart, muscle, skeleton, pigment and viscera. Embryonic cardiac arrhythmia caused by EE2 is probably resulted from heart abnormal structure. The embryonic stage sensitive to EE2 mainly started at cleavage and last up to the organogenesis with time-accumulating effect. RT-PCR results indicate that EE2 treatment disturbed gene expression pattern at the early period of zebrafish embryonic development by suppressing transcription of gene boz that promotes brain development, upregulating genes for trunk and tail, such as ntl, spt, shh, and perturbing Nodal signal expression of TGFbeta superfamily, for example, cyc, sqt and oep. Using zebrafish, an efficient in vivo method for quick evaluation of EE toxicity on embryonic development has been developed.

The paper reports the systematic study on felodipine and its impurities in tablets, to improve its quality standards for the control of the related substances. HPLC-DAD, UPLC-MS, IR and NMR methods were used for the isolation of felodipine and its impurities in tablets, their identification and the zebrafish animal model was used for the analysis of the toxic impurities. In felodipine material and its tablets, three impurities are isolated and identified. They are impurity 1 [dimethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], impurity 2 [ethyl methyl 4-(2, 3-dichlorophenyl)-2, 6-dimethylpyridine-3, 5-dicarboxylate] and impurity 3 [diethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], separately. The result of zebrafish animal model analysis showed that the teratogenic effects of four compounds were: impurity 3 > or = felodipine > impurity 1 > impurity 2, lethal effects were as follows: impurity 2 = impurity 3 > felodipine > or = impurity 1. This study confirmed the toxicity of three impurities in felodipine. According to the results, the paper suggested the amendments to the standard of the medicine and provided the support to the control of impurities in the manufacturing process.

Objective:To determine the expression of the full-length (NK1R-FL) and truncated (NK1R-Tr) neurokinin-1 receptor (NK1R) and the neurokinin-2 receptor (NK2R) in breast cancer tissues and cell lines, as well as to study the effects of the NK1R and NK2R antagonists on the growth of breast cancer cells. Methods:Immunohistochemistry and Western blot assays were used to detect NK1R, NK1R-FL, and NK2R expression in clinical samples of primary breast cancer tissue, benign lesions, and normal breast tissue, as well as in different breast cancer cell lines. Cell proliferation and soft agar growth tests were performed on cells treated with the NK1R and NK2R antagonists to study the ectopic overexpression of NK1R-FL and NK1R-Tr in breast cancer cell lines. Results:Total NK1R expression was detected in the breast cancer tissues, benign lesions, and normal breast tissues. Compared with the normal breast epithe-lia and benign breast lesions, the expression levels of NK1R-FL and NK2R decreased in the carcinoma. These changes were also relat-ed to the carcinoma type, histological grade, lymph node metastasis, HER2 and Ki-67 expression, and estrogen and progesterone recep-tors in breast cancer. The expression levels of NK1R-FL and NK2R were high in the HBL-100 breast cell lines of para-neoplastic tis-sues, but NK1R-Tr expression was low. The MDA-MB-231, T-47D, and MCF-7 cells only expressed NK1R-Tr. NK1R-Tr or NK1R-FL overexpression caused the decreased inhibition rate or increased levels of the NK1R and NK2R antagonists in the breast cancer cells. Conclusion:NK1R-FL and NK2R are co-expressed in normal cells. NK1R-Tr is highly expressed in breast cancer cells and exerts nega-tive feedback to regulate NK1R-FL and NK2R expression in all cells, especially cancer cells.

Objective To explore the correlation between plantar pressure and walking function in hemiplegic stroke patients.Methods Thirty hemiplegic patients with stroke (a hemiplegic group) and thirty age-matched healthy persons (a control group) were recruited.Gait and balance function training and assessment system (model:AL-600) were used to quantify the walking velocity,peak plantar pressure at heel-strike and push-off periods and displacement of center of pressure (DCOP) of all subjects during walking.The asymmetry of gait was calculated.Two independent sample t-test were used to compare the walking velocity,peak plantar pressure and DCOP for the two groups.Pearson correlation coefficients were applied to analyze the correlation between the walking velocity and peak plantar pressure and DCOP.Results The walking velocity,the peak plantar pressure at heel-strike and push-off periods and DCOP of the hemiplegic group were significantly lower than the control group.In the hemiplegic group,the asymmetry of peak plantar pressure and DCOPx significantly increased,while that of DCOPy became bigger without significant difference.Moreover,the walking capacity of the hemiplegic group was positively correlated with the peak plantar pressure and DCOP.Conclusion Among hemiplegic stroke patients,both the peak plantar pressure at heel-strike and push-off periods lower in a way.Their capacity of weight transfer decreases,which is closely related to their walking velocity.

Objective To explore the efficacy and safety of HAA regimen (homoharringtonine,cytarabine and aclarubicin) in the treatment of 150 newly diagnosed adult acute myeloid leukemia (AML).Methods All patients entered the study from May 1999 to June 2008 were treated with HAA regimen. Coxsurvival analysis was used to estimate the survival rate and differences between M1/M2 and M4/M5 were compared with 2-sided log-rank test. Results Out of the 150 patients, 121 (81%) achieved complete remission (CR). After the first course, CR rate was 68%. The CR rates of 97%, 84% and 38% were achieved in patients with favorable, intermediate and unfavorable cytogenetics, respectively. For the patients with CR, the median follow-up time was 16.5 ( 1.5-100.5 ) months, and the estimated 3-year survival rate was 45%. The estimated 3-year relapse free survival rate was 52% for the 121 patients with CR.Conclusions HAA regimen may be an efficacious and safe regimen with a good toleration in the induction therapy for newly diagnosed AML, and a high CR rate could be achieved with only one or two courses.

The paper reports the systematic study on felodipine and its impurities in tablets, to improve its quality standards for the control of the related substances. HPLC-DAD, UPLC-MS, IR and NMR methods were used for the isolation of felodipine and its impurities in tablets, their identification and the zebrafish animal model was used for the analysis of the toxic impurities. In felodipine material and its tablets, three impurities are isolated and identified. They are impurity 1 [dimethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], impurity 2 [ethyl methyl 4-(2, 3-dichlorophenyl)-2, 6-dimethylpyridine-3, 5-dicarboxylate] and impurity 3 [diethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], separately. The result of zebrafish animal model analysis showed that the teratogenic effects of four compounds were: impurity 3 > or = felodipine > impurity 1 > impurity 2, lethal effects were as follows: impurity 2 = impurity 3 > felodipine > or = impurity 1. This study confirmed the toxicity of three impurities in felodipine. According to the results, the paper suggested the amendments to the standard of the medicine and provided the support to the control of impurities in the manufacturing process.
Abnormalities, Drug-Induced ; Animals ; Antihypertensive Agents ; administration & dosage ; chemistry ; toxicity ; Calcium Channel Blockers ; administration & dosage ; chemistry ; toxicity ; Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Felodipine ; administration & dosage ; chemistry ; toxicity ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Pharmaceutical Preparations ; analysis ; chemistry ; Quality Control ; Spectrophotometry, Infrared ; Tablets ; Tandem Mass Spectrometry ; Zebrafish

External snapping hip(ESH) is a vague term used to describe palpable or auditory snapping with hip movements with or without pain. The pathogenesis of ESH is related to the specific anatomical structure and friction factor. The clinical symptom is auditory snapping during activities, physical examination, X-ray, magnetic resonance imaging(MRI), dynamic ultrasound and other imaging techniques can be used to diagnose. Conservative medical management includes rest, avoidance of aggravating activities, and antiinflammatory medications. Treatment Patients with mild symptoms can achieve good results by medication, rest and physiotherapy. Surgical treatment for patients with ineffective conservative treatment was performed. All kinds of open surgery method can achieve good clinical curative effect, arthroscopic surgery is gradually been promoted due to small trauma, less complications. Besides, there are some reports that traditional treatments such as massage, acupuncture and acupotomology have achieved good clinical results, which deserve further study and promotion.

OBJECTIVETo study the effects of glial cell derived neurotrophic factor (GDNF) on regeneration of facial nerve defects by autogenous facial vein conduit.

METHODSThirty-six rabbits were used in this study and 10 mm-length facial nerve defects were made on both sides of all animals. The nerve gaps were bridged using autogenous posterior facial vein graft of the same side. The animals received injection of either saline (group A, n=16) or GDNF (group B, n=16) into the veins. Nerve function was evaluated by evoking nerve action potential immediately after operation and 4, 8 and 16 weeks after operation. Regenerated nerve samples were harvested at 4, 8, and 16 weeks after operation and processed for histology and transmitting electron microscopic examination (TEM).

RESULTSAction potential did not exist immediately after operation but it was evoked at 4, 8, and 16 weeks in both groups. At 4 and 8 weeks after operation, the amplitude and width of action potential were significantly higher in group B than group A (P < 0.01), except wave width at 4 weeks, which showed no significant differences, while the latency period was significantly shorter in group B than that in group A (P < 0.01). At 16 weeks, action potential was similar between two groups, except wave amptitude, which was higher in group B than group A (P < 0.01). Morphologic and TEM examinations showed more matured myelinated nerve fibers and active Schwann's cells in group B when compared group A during the whole regeneration process.

CONCLUSIONGDNF can promote nerve regeneraat early stage during reconstruction of facial nerve defects by autogenous facial vein conduit and combination of GDNF and autogenous vein graft provides a valuable method for clinical reconstruction of facial nerve defects.

Animals ; Facial Nerve ; Nerve Growth Factors ; Nerve Regeneration ; Neuroglia ; Rabbits ; Regeneration

The pharmaceutical ethynylestradiol (EE) is a potent endocrine modulator. Application enlargement of ethynylestradiol in clinics and abuse in livestock farming and fishing make it important to explore ethynylestradiol toxicological action on vertebrate embryonic development and to establish an in vivo method for EE toxicity detection efficiently and conveniently. In the present study, using a model animal zebrafish and 17alpha-ethynylestradiol as a representative compound, we have investigated EE2 teratogenicity, target tissues and target genes on zebrafish embryo. The results show that median teratogenesis concentration (TC50) of EE2 is 0.8 microg x mL(-1), and median lethal dose (LD50) is 3.3 microg x mL(-1). Targets of EE2 action were implicated in brain, eyes, heart, muscle, skeleton, pigment and viscera. Embryonic cardiac arrhythmia caused by EE2 is probably resulted from heart abnormal structure. The embryonic stage sensitive to EE2 mainly started at cleavage and last up to the organogenesis with time-accumulating effect. RT-PCR results indicate that EE2 treatment disturbed gene expression pattern at the early period of zebrafish embryonic development by suppressing transcription of gene boz that promotes brain development, upregulating genes for trunk and tail, such as ntl, spt, shh, and perturbing Nodal signal expression of TGFbeta superfamily, for example, cyc, sqt and oep. Using zebrafish, an efficient in vivo method for quick evaluation of EE toxicity on embryonic development has been developed.
Abnormalities, Drug-Induced ; etiology ; Animals ; Arrhythmias, Cardiac ; chemically induced ; embryology ; Dose-Response Relationship, Drug ; Embryo, Nonmammalian ; abnormalities ; drug effects ; Embryonic Development ; drug effects ; Ethinyl Estradiol ; administration & dosage ; toxicity ; Gene Expression Regulation, Developmental ; Teratogens ; toxicity ; Zebrafish ; abnormalities ; embryology

OBJECTIVETo induce primary chronic myeloid leukemia (CML) cells into dendritic cells (DCs).

METHODSBone marrow mononuclear cells (MNCs) were isolated from 13 CML patients and peripheral blood MNCs from 5 healthy donors. The isolated MNCs were co-cultured with rhGM-CSF 1,000 U/ml, rhIL- 4,500 U/ml and TNF-alpha 50 U/ml for 10 days. The morphological features were observed by Wright's staining,inverted microscope and electron microscope. CD(80), CD(86), CD(83), CD(1a) and HLA-DR expression were assayed by flow cytometry, cytogenetic analysis was performed by fluorescence in-situ hybridization(FISH). The concentration of IL-12 was measured by ELISA and the function of antigen presenting was tested by mixed lymphocyte reaction (MLR).

RESULTAfter being cultured with cytokines, the typical dendritic appearance with delicate membrane projections was observed. The CD(80), CD(86), CD(83), CD(1a) and HLA-DR markers and capacity of stimulating allogeneic T cells were upregulated significantly. FISH confirmed that the DCs were generated from leukemic origin and CML DCs could secrete higher level of IL-12 than CML MNCs. There were no differences in morphology and immunophenotype expression between DCs derived from CML and those from normal individuals. However, DCs from CML patients displayed weaker activity than that of normal individuals when tested in MLR.

CONCLUSIONCML cells could be induced into leukemia-DCs by co-culture with cytokines.

Bone Marrow Cells ; immunology ; pathology ; Cell Differentiation ; Dendritic Cells ; cytology ; immunology ; Humans ; Interleukin-12 ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; pathology ; Tumor Cells, Cultured