Objective To study the receptors for the axon guidance factor Netrin-4 and whether the known Netrin-1 receptors, DCC and UNC5H1, are coreceptors for Netrin-4. Methods The receptor for Netrin-4 was detected by affinitycytochemistry and in situ staining with the alkaline phosphatase (AP) tagged Netrin-4 fusion protein. The dynamic character of the binding between Netrin-4 and its receptor was evaluated through ligand-receptor binding assay and competitive binding assay. Results In situ staining of AP4-Netrin-4 protein bound to COS7 cells transiently transfected with eukaryotic expression coding DCC and UNC5H1 showed that the Netrin-4 could bind to the transfected cells. Competitive binding assays showed that Netrin-1 could compete with Netrin-4 for the same receptors. Conclusion DCC and UNC5H1 are coreceptors for Netrin-4. However, because the binding affinity of Netrin-4 with its receptor was lower than that of Netrin-1,it could be inferred that there should exist unique receptor(s) for Netrin-4 other than DCC and UNC5H1.

Objective:To investigate the clinical values of touch imprint cytology (TIC)plus rapid immunohistochemistry staining (RIHC)in the intraoperative assessments of breast cancer sentinel lymph nodes (SLNs).Methods:A total of 281 SLNs from 89 patients of breast cancer were examined in this study.All those SLNs were evaluated with intraoperative touch imprint cytology and rapid immunohistochemistry staining.The RIHC indicator was pan-cytokeratin 1 9 (pan-CK 1 9 ).The two intraoperative methods are compared the results of SLNs metastases by postoperative routine pathological examination.Results:The detection rate of SLNs with TIC was 25.8% (23/89 );the detection rate with touch imprint cytology plus rapid immunohistochemistry staining technique was 36.0%(32/89),which was much higher than touch imprint cytology(P<0.05).The specificity was both 100.0%.The sensitivity of TIC and TIC plus RIHC for detecting positive lymph node was 69.7% (23/33)and 97.0% (32/33),respectively.The sensitivity of TIC plus RIHC was significantly higher than that of TIC (P<0.001).The false negative rate of SLNs with TIC was 30.3%(10/33);the false negative rate with TIC plus RIHC was 3.0%(1/33),which was lower than TIC (P<0.001).Conclusions:Touch imprint cytology plus rapid immunohistochemistry staining can provide a reliable and precise result for the intraoperative assessment of breast cancer SLNs.

The different biological functions were studied in mouse bone marrow-derived endothelial progenitor cells isolated by differential time attachment to obtain the optimal adherent time in this study. Density gradient centrifugation-isolated bone marrow mononuclear cells were seeded on the fibronectin-coated dish. The 1-day cultured unattached cells were seeded on the second dish for 2 more days. Then unattached cells in the second dish were seeded on the third dish. The cells on 3 dishes were defined as 1-day adherent cells, 3-day adherent cells and 3-day unattached cells, respectively. After 20-day culture, the biological functions, such as the percentage of biomarkers, the ability of adhesion, and the ability of forming tubes in vitro were analyzed. The results showed that the percentages of positive CD34, FLK-1, and CD34/FLK-1 expressions in 1-day attached cells were significantly increased compared to those in the 3-day adherent or unattached cells (P < 0.01), which showed the strongest adhesion ability. The expression of eNOS in 1- or 3-day adherent cells was significantly higher than that in 3-day unattached cells (P < 0.01). The expression of VEGF in 3-day adherent cells was significantly higher than that in 1-day adherent cells or 3-day unattached cells (P < 0.01). These results suggest the biological functions of 1-day adherent cells are significantly stronger than that of 3-day adherent or unattached cells. VEGF expression in 3-day adherent cells is higher than that in 1-day adherent cells or 3-day unattached cells. The expression of eNOS in 1-day adherent cells or 3-day adherent cells is higher than that in 3-day unattached cells. The optimal adherent time to obtain mouse bone marrow-derived endothelial progenitor cells is 1-3 d.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Leukocytes, Mononuclear ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Nitric Oxide Synthase Type III ; metabolism ; Stem Cells ; cytology ; metabolism ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism

Adolescent ; Blood Cell Count ; Child ; Copper ; blood ; Electromagnetic Fields ; adverse effects ; Female ; Humans ; Iron ; blood ; Magnesium ; blood ; Male ; Students ; Trace Elements ; blood ; Zinc ; blood

OBJECTIVETo investigate molecular mechanism of tanshinone II A inducing differentiation and apoptosis in acute promyelocytic leukemia NB4 cells.

METHODNB4 cells were cultured in vitro and treated with tanshinone II A and observed cellular morphology, cell category and the cellular proliferation. DNA microarray technique was used to analyze the gene expression profiles of NB4 cells induced by tanshinone II A.

RESULT92.8% of NB4 cells treated with 0.5 mg x L(-1) tanshinone II A were induced into mature neutrophils, in which myetocytes and melamyetocytes were 27.0%, banded and segmented neutrophits 68.2%. Cell growth were inhibited. cDNA microarray showed the enormously expressed 183 genes including 23 differentiation associated genes, and other interrelated genes.

CONCLUSIONTanshinone II A inducing differentiation in NB4 cells may be via regulation of many kinds of genes, especially differentiation associated genes expression. This partially explained the molecular mechanism of tanshinone II A inducing differentiation.

Cell Differentiation ; drug effects ; Cell Line, Tumor ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; genetics ; metabolism ; Phenanthrenes ; pharmacology

Cardiac Catheterization ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Prosthesis Implantation ; instrumentation ; Septal Occluder Device

AIMTo study the effect of CGRP receptor antagonist CGRP8-37 on nociceptive response and expression of nitric oxide synthase (NOS) and content of nitric oxide (NO) in the dorsal horn of the spinal cord of rats during formalin-induced inflammatory pain.

METHODSUsing formalin injection into right hind paw induced inflammatory pain. Counting the times of flinching reflex was used to observe the degree of spontaneous pain. NADPH-d histochemistry was used to observe the changes of NOS expression. The content of NO was observed by measuring the contents of nitrate/nitrite (NO3- / NO2-).

RESULTSspontaneous pain behavioral was elicited by formalin injection. The NOS expression and NO content significantly increased in the spinal cord at 24 h after formalin injection. Intrathecal injection of CGRP8-37 could significantly inhibit the response of spontaneous pain and the increases of NOS expression and NO content induced by formalin injection.

CONCLUSIONThe activation of CGRP receptors enhances NOS expression and NO production in the dorsal horn of the spinal cord during formalin-induced inflammatory pain.

Animals ; Calcitonin Gene-Related Peptide ; pharmacology ; Formaldehyde ; adverse effects ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Pain ; chemically induced ; metabolism ; Peptide Fragments ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitonin Gene-Related Peptide ; antagonists & inhibitors ; Spinal Cord ; drug effects ; metabolism

Four impurities were isolated from raw material of clindamycin phosphate (CP), and their structures have been determined. LC-MS was used to determine the molecular weights of the impurities in the raw material of CP. Reversed-phase preparative HPLC was used to prepare them, and their chemical structures were identified by HR-MS and NMR. The four unknown impurities were determined as clindamycin-B-phosphate (1), clindamycin-2,4-diphosphate (2), 3',6'-dehydro clindamycin phosphate (3), epi-clindamycin phosphate (4). Impurity 1 has been included in BP and EP, while 2, 3 and 4 have not. The impurities 2, 3, 4 are first separated from raw material of CP.
Anti-Bacterial Agents ; chemistry ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Clindamycin ; analogs & derivatives ; chemistry ; Drug Contamination ; Molecular Structure ; Molecular Weight ; Spectrometry, Mass, Electrospray Ionization

Objective To study the correlation between the method Strengthening Earth to Generate Metal ( SEGM ) on infantile repeated re-spiratory infection ( RRTI) of lung and spleen qi deficiency and the annual incidence of respiratory tract as well as the correlated immune response.Methods After the case screening , the selected patients were divide into 4 groups: control group (17 cases, no drugs), Chinese medicine group (36 cases, Shenling Baizhu particles , 1.5 -3 g each time, 2 times a day), western medicine group (36 cases, pidotimod granules, 0.4 g each time, 2 times a day) and integrated group of traditional and western medi-cine (37 cases, Shenling Baizhu granule plus Pidotimod granules ).The changes of children′s annual incidence and the immune index before and after treatment for 8 weeks were observed.Results The method of strengthening earth to generate metal can reduce the annual incidence of repeated respiratory tract infection ( P<0.05 ) , and can regulate the T lymphocyte subpopulation homeostasis , with serum immunoglobulin levels elevated (P<0.05).Conclusion The method of SEGM can reduce the annual incidence in chlid patients with recurrent respiratory tract infection of lung and spleen qi deficiency ( P<0.05);and regulate the immunity of the patients.

OBJECTIVETo evaluate the efficacy, side-effects and quality of life in the advanced colorectal cancer patients treated by irinotecan plus fuorouracil and leucovorin with thalidomide or without thalidomide.

METHODSEligible patients were randomly assigned to the treatment group and control group in a 1:1 ratio. In the treatment group, 32 evaluable patients were treated with irinotecan 180 mg/m2 i. v. on day 2, fuorouracil 400 mg/m2 bolus on day 1, 2 at a dose of 1200 mg/m2 civ. for 43 hours; leucovorin 200 mg/m2 i. v. on day 1, 2; thalidomide 300 mg, orally on day 1 - 14, two weeks as a cycle. In the control group, the regimen was the same as in the treatment group except oral intake of thalidomide.

RESULTSThe response rate was 28.1% in the treatment group vs. 15.2% in the control group (P = 0.2034) with a median TTP of 3.8 months vs. 2. 5 months (P = 0.1312). Furthermore, there was no statistically difference either between two groups regarding to adverse effects.

CONCLUSIONIrinotecan plus fuorouracil and leucovorin without oral intake of thalidomide is as effective and tolerable as irinotecan plus fuorouracil and leucovorin combined with oral thalidomide for advanced colorectal cancer.

Adenocarcinoma ; drug therapy ; Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Camptothecin ; administration & dosage ; analogs & derivatives ; Colorectal Neoplasms ; drug therapy ; Diarrhea ; chemically induced ; Disease Progression ; Female ; Fluorouracil ; administration & dosage ; Humans ; Leucovorin ; administration & dosage ; Male ; Middle Aged ; Remission Induction ; Survival Analysis ; Thalidomide ; administration & dosage ; Treatment Outcome