Objective To study the effects of prenatal cocaine exposure on the development of offspring's brains by building a murine model. Methods We weighted the body weight and brain weight of offspring on P10 from COC and SAL groups and observed the development of neuron and astrocyte in cerebral cortex by toluidine blue staining and immunohistochemistry. Results The brain weight and body weight from COC were both reduced on P10 compared with SAL group.We discovered prenatal cocaine exposure induced polarity disorder and dysplasia of neuron in cerebral cortex;the number of the astrocytes in corpus callosum and hippocampus regions decreased.Conclusion\ Pregnatal cocaine exposure can result in abnormal development of cerebral cortex of offsprings which may play an important role on cocaine induced abnormal behavior.\;[

AIM To develop a murine model for investigating the effects of prenatal cocaine exposure on the development of fetal nerve system. METHODS A nutritionally paired control group of dams injected with saline and pair fed with the COC dams were set up. Another two groups were COC groups injected with cocaine HCl and SAL group administrated with saline. After injection twice daily during gestation days 8～17,mice were decapitated on E17 and blood and brain samples were collected for pharmacological analysis and neurotransmitter analysis by HPLC.RESULTS Pharmacological analysis revealed that cocaine was found in maternal and fetal plasma at 15 min following ip administration to embryonic day E17 pregnant mice. Though COC dams and SPF dams had the same feeding condition, compared with the latter, the former had higher maternal concentrations of DA and 5 HT, lower fetal weight, brain weight, striatum weight and higher concentrations of DA and 5 HT in striatum, P

AIM To investigate the effects of in utero cocaine exposure on the fetal development, when fetuses were exposed to equal total dose but different dose and timing. METHODS Pregnant dams were randomly separated into three groups: SAL, COC20 and COC40. On E17, recorded body weight, brain weight and striatum weight of all groups, and examined the concentrations of DA and 5 HT in fetal striatum by HPLC. RESULTS Body weight of SAL, COC40, COC20 groups decreased progressively in turns. Brain weight of COC20 group and COC40 group was lower than that of SAL. Only the brain/body ratio of COC40 was decreased ( P

The aim of the experiments was to develop and characterize a murine model for investigating the effects of prenatal cocaine exposure on the mother and fetus. Pregnant mice were separated into three groups: group 1 was treated with cocaine HCl at 10 mg/kg twice daily (COC); group 2 was treated with saline at 10 ml/kg twice daily (SAL); and group 3 was pair-fed with the COC dams and was injected with saline following the same schedule (SPF) from embryonic day (E) 8 to 17. We utilized high-pressure liquid chromatography (HPLC) with UV detector and electrochemical detector to test the concentrations of cocaine, dopamine and serotonin, as well as HE staining to observe morphological alterations of liver and placenta. Though less food intake and lower weight gain were observed in COC and SPF groups but not in SAL dams, lower fetal body weight and brain weight were only seen in COC offspring. Pharmacological analysis revealed that cocaine was found in fetal plasma at 15 min following intraperitoneal administration on E17, accompanied with elevated concentrations of dopamine (DA) and serotonin (5-HT) in fetal brain. We also observed morphological changes in liver and placenta of cocaine-exposed fetuses. The present study indicates that pregnancy cocaine exposure can lead to maternal undernutrition and developmental abnormality of the fetal brain, liver and placenta. It is suggested that the developmental abnormality of the fetuses induced by cocaine is due to the toxicological effect of cocaine but not to maternal undernutrition.
Animals ; Brain ; metabolism ; pathology ; Cocaine ; adverse effects ; blood ; Disease Models, Animal ; Dopamine ; metabolism ; Female ; Fetus ; drug effects ; pathology ; Liver ; pathology ; Malnutrition ; Maternal Exposure ; adverse effects ; Maternal Nutritional Physiological Phenomena ; Mice ; Mothers ; Placenta ; pathology ; Pregnancy ; Serotonin ; metabolism

OBJECTIVE: To investigate the inhibition effects of Tianshen Yizhi Recipe (TSYZR), a compound traditional Chinese herbal medicine, on decreased expression of nicotinic acetylcholine receptor (nAChR) and the neurotoxicity as well as lipid peroxidation induced by beta-amyloid peptide (Abeta) in human SH-SY5Y neuroblastoma cells. METHODS: The SH-SY5Y cells were treated by a certain concentration of TSYZR, and then exposed to Abeta(25-35). Methyl thiazolyl tetrazolium reduction assay was carried out to understand the influences of the drugs on cellular viability. Expressions of nAChR subunits (alpha3 and alpha7) at protein and mRNA levels were detected by Western-blotting and reverse transcription polymerase chain reaction, respectively. Lipid peroxidation was measured by thiobarbituric acid to observe the capacity of antioxidant of the drugs. RESULTS: TSYZR at a safe concentration could increase alpha7 protein in the cells, inhibit decreased expressions of alpha3 and alpha7 nAChR subunit proteins, prevent lower expression of alpha7 mRNA in SH-SY5Y cells induced by Abeta, reduce the neurotoxicity and lipid peroxidation resulting from Abeta, but had no significant effect on the lower expression of alpha3 mRNA. CONCLUSIONS: TSYZR can up-regulate the expression of alpha7 nAChR subunit protein and prevent decreased expressions of nAChRs and neurotoxicity as well as lipid peroxidation induced by Abeta. This drug may play an important therapeutic role in treatment of Alzheimer disease.

AIMTo develop a high throughput screening assay to identify inhibitors of intercellular adhesion molecule-1 (ICAM-1) expression in human umbilical vein endothelial cells (HUVEC).

METHODSICAM-1 expression in lipopolysaccharide-stimulated endothelial cells was measured by ELISA. The cytotoxicity of the compounds was measured by MTT.

RESULTSLipopolysaccharide (LPS) increased ICAM-1 expression in HUVEC in a concentration- and time-dependent manner. Two thousand compounds were screened and the hit rate was 1.5%. Among these 30 compounds, 24 were cytotoxic.

CONCLUSIONThe ELISA method was inexpensive, reproducible and suitable for high throughput primary cell assay. This assay was feasible to identify inhibitors of ICAM-1 and simultaneously discriminate the activity from the cytotoxic effects.

Cell Adhesion Molecules ; antagonists & inhibitors ; biosynthesis ; Cells, Cultured ; Chemistry, Pharmaceutical ; methods ; Drug Evaluation, Preclinical ; methods ; Endothelium, Vascular ; cytology ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; drug effects ; metabolism ; Lipopolysaccharides ; pharmacology ; Umbilical Veins ; cytology ; metabolism

BACKGROUNDLipoprotein-associated phospholipase A2 (Lp-PLA2) is mainly secreted by macrophages, serving as a specific marker of atherosclerotic plaque and exerting pro-atherogenic effects. It is known that high-density lipoprotein (HDL) plays an important role against atherosclerosis by inhibiting pro-inflammatory factors, however, the relationship between HDL and Lp-PLA2 remains elusive.

METHODSIn this study, reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and a platelet-activating factor (PAF) acetylhydrolase assay were performed to determine the Lp-PLA2 mRNA level, protein expression and activity in human monocyte-derived macrophages upon HDL treatment of different concentrations and durations. To investigate the underlying mechanism of HDL-induced Lp-PLA2 action, pioglitazone, a peroxisome proliferator-activated receptor-γ (PPARγ) ligand, was introduced to human monocyte-derived macrophages and mRNA and protein levels of Lp-PLA2, as well as its activity, were determined.

RESULTSLp-PLA2 mRNA levels, protein expression and activity were significantly inhibited in response to HDL treatment in a dose and time dependent manner in human monocyte-derived macrophages. Pioglitazone treatment (1 - 10 ng/ml) upregulated the Lp-PLA2 mRNA level, protein expression and activity in human monocyte-derived macrophages, while the effects were markedly reversed by HDL. In addition, pioglitazone resulted in a significant increase in PPARγ phosphorylation in human monocyte-derived macrophages, which could be inhibited by HDL.

CONCLUSIONThese findings indicate that HDL suppresses the expression and activity of Lp-PLA2 in human monocyte-derived macrophages, and the underlying mechanisms may be mediated through the PPARγ pathway.

1-Alkyl-2-acetylglycerophosphocholine Esterase ; genetics ; metabolism ; Cells, Cultured ; Humans ; Lipoproteins, HDL ; pharmacology ; Macrophages ; drug effects ; enzymology ; metabolism ; PPAR gamma ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; genetics

Objective: This study was designed to investigate the prognostic value of serum β2-micorglobulin(β2-MG) level in non-Hodgkin s lymphoma(NHL) patients and its correlation with Ann Arber staging system and the pathological type. Method: Serum β2-MG levels in 75 de novo NHL patients were measured by radioimmunoassay. Results: The β2-MG level in 75 NHL patients was higher than that in nomal control level in 52％ of the patients and correlated with the Ann Arber stage. There was significant difference between two groups (P<0.01). The international prognostic index (IPI) and the response rate to chemotherapy were also correlated with the β2-MG level in serum. The response rate, the complete response, and partial response were 87.2%, 61.5%, and 25.6% in normal serum β2-MG group, while in the high β2-MG group they were only 75.0%, 50.0%, and 25.0%, respectively, which were significantly different between the two groups (P<0.05). The difference of 5-year survival between the two groups was significant (64.1% vs 27.8%, P<0.05) and favorable in normal β2-MG group. Conclusion: β2-MG level in serum is an independen prognostic factor in de novo NHL, and the patients with abnormal β2-MG level in serum have a unfavorable prognosis.

BACKGROUNDMolecular targeted drugs is now widely used in non-small cell lung cancer (NSCLC) clinical treatment. Icotinib hydrochloride is a new type of oral epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKIs). In this study, we examined the role of EGFR, K-RAS, B-RAF somatic mutations and EGFR mRNA expression in tumor specimens from advanced NSCLC patients as predicators of the efficacy of icotinib hydrochloride.

METHODSWe analyzed tumor paraffin-embedded specimens, which were obtained from 14 of 40 patients with advanced NSCLC who enrolled in the stage I clinical trial of icotinib hydrochloride. Somatic mutations were evaluated by mutant-enriched liquidchip (MEL) technology, and EGFR mRNA expression was measured by branched DNA liquidchip (MBL) technology.

RESULTSIn the 14 specimens, seven patients showed EGFR mutations, exon 19 deletion (3/7) and exon 21 point mutation (4/7); and two patients showed K-RAS mutation. No mutations in EGFR exon 20 or B-RAF were detected. In patients with EGFR mutation, one patient developed progress disease (PD), three patients had stable disease (SD), two patients had partial responses (PR) and one patient had a complete response (CR). In patients with wild-type EGFR, four patients had PD, three patients acquired SD, and none had PR/CR (P = 0.0407). EGFR mutations were associated with better progress-free survival (PFS) (141 days vs. 61 days) but without a statistically significant difference (P = 0.8597), and median overall survival (OS) (≥ 449 days vs. 140 days). EGFR mRNA expression levels were evaluated (three high, eight moderate, one low, and two that can not be measured due to insufficient tumor tissue) and no statistically significant relationships was observed with response, PFS or OS.

CONCLUSIONSThe EGFR mutation rate was consistent with that reported in the Asian population, so the MEL technology is reliable for measuring EGFR mutation with high throughput and rapidity. EGFR exon 19 deletions and exon 21 point mutation are predictive biomarkers for response to icotinib hydrochloride as second line treatment or above.

Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; Crown Ethers ; therapeutic use ; Exons ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Proto-Oncogene Proteins B-raf ; genetics ; Proto-Oncogene Proteins p21(ras) ; genetics ; Quinazolines ; therapeutic use ; RNA, Messenger ; genetics ; Receptor, Epidermal Growth Factor ; genetics

BACKGROUNDA retrospective analysis of clinical data were conducted reviewing patients who were given erlotinib at Peking Union Medical College (PUMC) Hospital from May 2005 to December 2009. Relationships between clinical factors, epidermal growth factor receptor (EGFR) mRNA expression, EGFR gene mutations, KRAS gene mutations and clinical outcomes were investigated in Chinese patients with advanced non-small cell lung cancer (NSCLC).

METHODSPatients with stage IIIB/IV NSCLC who had not previously participated in erlotinib related clinical trials were enrolled into this study. All patients were given oral erlotinib 150 mg per day. Tumor samples of some patients were accessed with mutant-enriched polymerase chain reaction assay (EGFR, KRAS gene mutations) and multiplex branched DNA assay (EGFR mRNA expression).

RESULTSSeventy-nine patients were enrolled in this study, 23 patients had a partial response (PR), 36 patients had a stable disease (SD), 20 patients had a PD, with an objective response rate of 29.1%, and a disease control rate of 74.7%. Females (P = 0.023), non-smokers (P = 0.013), patients with a skin rash (P = 0.047), and with highly differentiated tumors (P = 0.037) were significantly correlated with the objective response rate. Patients with a lower ECOG PS (P = 0.002), highly differentiated tumors (P = 0.014), non-smokers (P = 0.002), and patients with a skin rash (P < 0.001) were significantly correlated with the disease control rate. The median progression-free survival was 35 weeks (95%CI: 13 - 57 weeks) and 1-year survival was 72.3%. Highly-differentiated tumors (P = 0.027) and patients with a skin rash (P < 0.001) were significantly correlated with PFS. Seventeen patients were tested for EGFR/KRAS gene mutations and EGFR mRNA expression. Progression-free survival (PFS) of patients with EGFR exon 19/21 mutations was 66 weeks, longer than patients with wild type EGFR exon 19/21 (P = 0.018). No significant relationships were found between EGFR mRNA expression, EGFR exon 19/21 mutations, and KRAS mutations and objective response rate or disease control rate. The most common adverse events were skin rash (60.9%) and diarrhea (26.6%).

CONCLUSIONSErlotinib was safe and effective in treating Chinese patients with advanced NSCLC. The PFS of patients who had a skin rash, highly differentiated tumors, or EGFR exon 19/21 mutations was significantly longer.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Erlotinib Hydrochloride ; Female ; Humans ; Male ; Middle Aged ; Protein Kinase Inhibitors ; therapeutic use ; Quinazolines ; therapeutic use ; Retrospective Studies ; Treatment Outcome