Objective To explore the change of degrade of clonazepam in serum with single and repeated - dose administration in children with seizure, and find a reasonable method for using the clonazepam. Methods Children with seizures were divided into single - dose paradigam, repeated - dose paradigam, and decreased - dose paradigam. The concentration of CZP in serum was determined by high performance liquid chromatography( HPLC). Results The serum concentrations of clanazepain in single - dose paradigam were (101.9?12.1),(76.9 ? 5.8),(50.7?2.9),(30.9?5.4),(21.5?6.8)?g/L,the time point that the blood samples collected were 15,30,60,120 and 480 min. The serum concentrations in repeated - dose paradigam were (97. 2 ? 6. 1),(130.4? 13. 4), (99. 4 ? 9.8),(79.6?2.4)?g/L,in decreased-dose paradigam were( 101.1 ?13.1),(123.1 ?6. 6), (99.4 ?9. 8), (79. 3 ? 2. 2)?g/L,in these two groups,the time point were 15,45,60 and 120 min. Conclusion Repeated administration of CZP with decreased dose may increase its effectiveness in treatment without substantially increasing toxicity.

Aim To observe the expression of migration inhibitory factor (MIF) in the enteric neurons,and to explore the nervous regulation on MIF activity in experimental colitis.Methods Colitis was induced in sensitized rat and mouse by 2,4-dinitrochlorobenzene(DNCB)enema.MIF activity was measured both in the mesentery lymphocyte(by MTT)and in the enteric neurons(by immunofluorescence double staining).6-OHDA was intraperitonealy (ip) administered to mouse before DNCB treatment.Norepinephrine(NE) was added to lymphocyte culture in vitro during MIF preparation.Results The expression of MIF protein in enteric neuron was increased in DNCB-induced colitis in rat.ip 6-OHDA in colitis mouse(38～150 mg?kg-1) resulted in a further increase of MIF activity than ip vehicle in colitis mouse (P

0.99.The quantitive restriction was 0.1 ?g/mL,the recovery rate was over 70%.The RSD of intra-and inter-day was less than 15%.The pharmacokinetic parameters of flavonoids were different from each other after ig and iv administration.The absolute bioavailability of flavonoids were 12.9%,10.8%,2.2%,10.2% and 5.9%,respectively.Conclusion The method is sensitive,specific,accurate,and is not only useful for guiding the bioavailability study on the corolla of A. manihot,but also for establishing a reasonable clinical dosage program.The four flavonols in the corolla of A.manihot can be rapidly distributed and eliminated in rats,the pharmacokinetics of two routes of administration are different.A novel integrated pharmacokinetic approach to describing the holistic pharmacokinetic properties of Chinese materia medica has been successfully developed and validated using four flavonols of A.manihot as a model herbal medicine.This study would be a new try for guiding the holistic pharmacokinetic study in consistence with the intrinsic theory and characteristics of traditional Chinese medicine.

Objective To study the expression and clinical significance of CD34 and smooth muscle actin (SMA) in stroma of breast tissues and lesions. Methods Seventy cases of breast tissues and lesions, including 20 fibroadenomas, 10 sclerosing adnoses, 30 invasive ductal carcinomas and 10 invasive lobular carcinomas were investigated, and 10 normal breast tissues were served as controls. Immunohistochemical staining was applied to compare the distribution of CD34+ fibrocytes and SMA-reactive myofibroblasts. Results The stroma of normal breast tissue contained CD34+ fibrocytes, whereas SMA-reactive myofibroblasts were absent (100% for both). All benign breast lesions exhibited astromal CD34+ fibrobytes, and fibroadenomas showed SMA-reactive myofibroblasts as well. In invasive ductal and lobular carcinoma the stroma was devoid of CD34+ fibrocytes, but a varying number of stromal SMA-reactive myofibroblasts were detectable (100%). Conclusion In breast carcer, immunohistochemical staining used in detecting expressions of CD34 and SMA is helpful in distinguishing benign lesions from malignancies.

To study the effect of miR-490 on Coxsackievirus B3 (CVB3) replication, HeLa cells were trans- fected with miR-490 in vitro, and infected with a Renilla luciferase (RLuc)-expressing CVB3 variant (RLuc-CVB3). The activities of RLuc in these cells were measured at 8h intervals from 0 to 40 h post-infection (p.i.), and the effects of miR-490 on RLuc-CVB3 replication were observed. In a further study, HeLa cells were transfected with either miR-490 or antisense miR-490 (AMO-miR-490), and were then infected with an enhanced green fluorescence protein (EGFP)-expressing CVB3 variant (EGFP-CVB3). The replication of EGFP-CVB3 was then determined by detecting the expression of EGFP. We observed that miR-490 could significantly inhibit the expression of RLuc in infected cells at 32 h p. i. Furthermore, in HeLa cells infected with EGFP-CVB3 at 32 h p.i., EGFP expression was also significantly inhibited by the presence of mniR-490. The inhibitory effect of miR-490 on EGFP expression in EGFP-CVB3-infected cells could be reversed by tranfection with AMQ-miR-490. These results indicated that miR-490 significantly inhibits the replication and expression of QVB3.
Enterovirus B, Human ; genetics ; physiology ; Enterovirus Infections ; genetics ; metabolism ; virology ; HeLa Cells ; Humans ; MicroRNAs ; genetics ; metabolism ; Virus Replication

Adolescent ; Adult ; Aged ; Cadherins ; genetics ; metabolism ; physiology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Ki-67 Antigen ; genetics ; metabolism ; Male ; Microtubule-Associated Proteins ; genetics ; metabolism ; physiology ; Middle Aged ; Neoplasm Invasiveness ; physiopathology ; Pituitary Neoplasms ; pathology ; physiopathology ; Young Adult

AIM: In order to explore the pathogenesis of ulcerative colitis (UC), an experimental colitis in mouse was induced by the hapten dinitrochlorobenzene (DNCB), and the activity of leukocyte migration inhibitory factor (LMIF) was measured at the same time. METHODS: 67 BALB/c mice were randomly divided into control (60% ethanol) and DNCB groups. After they were sensitized by smearing 3.3% DNCB on the abdominal skin, they were challenged with DNCB at concentration of 0.1%, 0.2% and 0.4% respectively by instillation once a day. The weight, stool viscosity and hematochezia were observed and accumulated as disease active index (DAI) score. The pathological changes in colon tissue were judged macropathologically and by means of microscope. LMIF activity was determined by the absorbance (A) of migrated leukocytes. RESULTS: Compared to control group, the increases in DAI accumulate score, pathologic score, and LMIF activity in DNCB groups were observed. CONCLUSION: Mouse colitis was induced by DNCB, which was accompanied by an increase in LMIF activity. [

OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alka-loids biosynthetic pathways, we conducted the transcriptome analysis of L. casuarinoides by illumina sequencing.METHODS The plant of L.casuarinoides was collected and was subjected to RNA isolation, cDNA library construction, and illumina sequencing before bioinformatics analysis. After sequencing, the clean reads were obtained for de novo assembly by using Trinity software,and then further processed with TGICL sequencing clustering software to generate unigenes,The unigenes are aligned by Blast X alignment to six public protein database. In addition, all unigenes are functionally annotated by GO, KEGG and characterized putative genes involved in lycopodium alkaloids biosynthesis. RESULTS In total, 124, 524 high-quality unigenes were obtained with an average sequence length of 601 bp. Among the L. casuarinoides transcripts, 61,304 showed significant similarity (E-value<1 e-5) to the known proteins in the public database.Among the total 124 524 unigenes,47,538 open reading frame (ORFs) were predicted. Based on the bioinformatics analysis, all possible enzymes involved in the Lycodine-type alkaloids biosynthetic pathway of L. casuarinoides were identified, including primary amine oxidase (PAO), and Malonly-CoA decarboxylase. In addition, a total of 64 putative cytochrome P450 (CYP450) and 827 transcription factors were selected as the candidates of Lycodine-type alka-loids modifiers. Furthermore, a total of 13 352 simple sequence repeats (SSRs) were identified from the 124, 524 unigenes, of which dinucleotide motifs AG/CT (50.1%), were the most abundant. CONCLU-SION This transcriptome analysis of L.casuarinoides,provides many valuable candidate genes involv-ing in the biosynthesis of novel secondary metabolites but also lays the foundation for genetic diversity analysis via SSRs markers in L.casuarinoides.

OBJECTIVETo explore the effect of ERK1/2 MAPK pathway on the expression of Kv1.5 channel, a voltage-gated potassium ion channel, in rat pulmonary artery smooth muscle cells (PASMCs) and its mechanisms during the process of hypoxia.

METHODSThe PASMCs derived from SD rats were cultivated primarily. The third to sixth generation of PASMCs were divided into 5 groups randomly: (1) Normal group (N); (2) Hypoxic group (H); (3) Demethy sulfoxide(DMSO) group (HD); (4) U0126 group (HU): 10 micromol/L U0126; (5) Anisomycin group (HA): 10 micromol/L anisomycin. There were three dishes of cells in each group. The cells in normal group were cultured in normoxic incubator (5% CO2, 37 degrees C), the cells in other groups were added to 0.05% DMSO in the hypoxic incubator (5% CO2, 2% O2, 37 degrees C), all cells were cultured for 60 h. RT-PCR and Western blot were used to detected the espressions of Kv1.5 mRNA and protein in PASMCs.

RESULTSCompared with N group, the expressions of Kv1.5 mRNA and protein in H, HD and HA groups were reduced significantly (P < 0.05); Compared with H group and HD groups, Kv1.5 mRNA and protein expressions in HU group were increased sharply (P < 0.05). Compared with the HU group, Kv1.5 mRNA and protein expressions in HA groups were significantly lower (P < 0.05).

CONCLUSIONLow oxygen reduced Kv1.5 mRNA and protein expressions, U0126 could resistant the Kv1.5 channel lower expression caused by hypoxia. Anisomycin had no significant effect on Kv1.5 channel expression under hypoxia, but the expression of Kv1.5 was still significantly lower than the normal oxygen group. These data suggest that hypoxia may cause hypoxic pulmonary hypertension by interfering ERK1/2 signaling pathway to inhibit Kv1.5

Animals ; Cell Hypoxia ; Hypertension, Pulmonary ; Kv1.5 Potassium Channel ; metabolism ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; Oxygen ; Pulmonary Artery ; cytology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley

Calcinosis ; pathology ; Child ; Humans ; Inhibins ; metabolism ; Male ; S100 Proteins ; metabolism ; Sertoli Cell Tumor ; metabolism ; pathology ; surgery ; Testicular Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism