Animals ; Cells, Cultured ; Cyclic AMP ; metabolism ; Pituitary Gland, Anterior ; cytology ; drug effects ; metabolism ; Rats ; Substance P ; pharmacology

Purpose To investigate the effect of subcellular location of tumor BRCA1 on the sensitivity to ionizing radiation (IR) and PARP inhibitor.Methods siRNA of BRCA1 were first used to inhibit endougenous BRCA1 expression in MCF7 cells.Then,plasmids of pCMV-3xFlag-WT-BRCA1,pCMV-3xFlag-NES-BRCA1 and pCMV-3xFlag-NLS-BRCA1 were transfected in MCF7 cells.Immunofluorescence staining was used to detect BRCA1 subcellular location as well as the formation of Rad51 and γ-H2AX foci.Apoptotic cells were analyzed by flow cytometry,and colony formation assay was performed to evaluate the survival of cells.Results There were 47％ cells with nuclear BRCA1,23％ cells with cytoplasmic BRCA1 and 30％ cell with mixed nuclear and cytoplasmic BRCA1 expression in WT-BRCA1 transfected cell.There were 87％ cells with nuclear BRCA1 in NES-BRCA1 transfected cell,and 82％ cells with cytoplasmic BRCA1 in NLS-BRCA1 transfected cell.There were 87％,84％ and 13％ Rad51 foci positive cells at 2 hours after 4 Gy radiation treatment and 22％,25％ and 59％ γ-H2AX foci positive cells at 24 hours after 4Gy radiation treatment in WT-BRCA1,NES-BRCA1 mutant and NLS-BRCA1 mutant transfected cell respectively.ABT-888 and radiation treatment induced more apoptosis and fewer colonies in NLS-BRCA1 transfected cell than WT-BRCA1,NES-BRCA1 mutant transfected cell.Conclusion Subcellular location of BRCA1 might affect homologous recombination repair of DNA double strand breaks and can be used to predict sensitivity to IR and PARP inhibitor.

Objective To explore the effects of polymorphisms of Crohn's disease related NOD2 gene and human beta-defensin 2 (hBD-2) on transcription of hBD-2 gene and its mechanism. Methods HEK293T cells were transfected with hBD-2 gene and NOD2 eukaryotic expression plasmid, and were then stimulated with LPS, TNF-α, or BAY 11-7082 (antagonist of NF-κB), respectively. Transcriptional activity of hBD-2 was detected afterwards. Results LPS could suppress transcription of hBD-2 (P=0. 020), which was increased by TNF-α in a dose-dependent manner (P =0. 004). In the presence of LPS, there was sig-nificant difference in transcriptional activity of hBD-2 between wild-NOD2 transfected group and mutated NOD2 (P268S) transfected group (P=0. 008), but there was no significant difference between wild hBD-2 transfected group and mutated hBD-2 transfected group (P=0. 053). With the stimulation of TNF-α (5 ng/ml), there was a significant difference between mutated hBD-2 transfected group and wild hBD-2 transfected group (P=0. 006), but no significant difference between wild-NOD2 transfected and mutated NOD2 transfected group was defected (P = 0. 064). Pretreatment with BAY 11-7082 before TNF-α (5 ng/ml) significantly inhibited the transcriptional activity of hBD-2 (P < 0. 001). Conclusion The poly-morphism of NOD2 affects the innate expression of hBD-2, the polymorphism of site in hBD-2 promoter (-233) may lead to significant decline of the inducible expression of hBD-2, and NF-κB might be a key pathway that NOD2 protein mediates the expression of defensin.

Objective To investigate the association of two single nuclear peptides(SNPs)polymorphisms(rs11209026 and rs11805303)which lies in interleukin-23 receptor(IL23R)gene with susceptibility to inflammatory bowel disease(IBD).Methods The target SNPs were directly sequenced by polymerase chain reaction(PCR)and gene polymorphisms of 50 healthy and 81 patients with IBD (Crohn's disease in 41 patients and ulcerative colitis in 40 patients)were analyzed using chromassoftware.Results The geno-type frequency and allelic frequency of rs11209026 were 7.3%and 3.7%in patients with Crohn's disease respectively,15.0%and 7.5%in patients with ulcerative colitis respectively as well as 14.0%and 7.0%in normal population respectively(all P value＞0.05).The geno-type frequency and allelic frequency of rs11805303 were 22.0%and 52.4%in patients with Crohn's disease respectively,15.0% and 41.2% in patients with ulcerative colitis respectively as well as 34.0%and 59.0%in normal population respectively(all P value＞0.05).But in allelic frequency there was significant difference between ulcerative colitis patients and normal population(P=0.018).The polymorphisms of rs11805303 loci did not correlate with age,gender,disease duration.activity and site in patients with ulcerative colitis.Conclusions IL23R gene polymorphism is not associated with the susceptibility to Crohn's disease.rs11805303 allele may be related with susceptibility to ulcerative colitis.But no correlation was found between the SNP polymorphisms and the clinical characteristic of ulcerative colitis.

OBJECTIVETo evaluate the clinical effectiveness of transurethral seminal vesiculoscopy (TUSV) combined with finasteride in the treatment of recurrent hemospermia.

METHODSThis study included 32 patients with recurrent hematospermia, with the disease course of 3 months to 4 years. After administration of finasteride at 5 mg/d for 2 weeks, the patients underwent TUSV for both exploration of the causes and treatment, followed by medication with finasteride at the same dose for another 2 weeks. Postoperative follow-up was conducted for observation of the outcomes and complications.

RESULTSTUSV was successfully accomplished in all the 32 cases, which revealed 16 cases of seminal vesiculitis, 10 seminal calculi, 1 seminal vesicle cyst, 2 seminal vesicle polyps, and 3 seminal vesicle abscess. The operative time was 20 to 51 (31.0 +/- 5.2) minutes. Postoperative complications included 1 case of acute epididymitis and 3 cases of breast discomfort within the first 4 weeks. No incontinence, urethral stricture, rectal injury, retrograde ejaculation, and sexual dysfunction occurred postoperatively. All the patients but 1 were followed up for 6 months to 2 years. Twenty-nine of the cases were cured, and 2 experienced recurrence.

CONCLUSIONTransurethral seminal vesiculoscopy combined with finasteride is safe and effective for the treatment of recurrent hemospermia.

Adult ; Endoscopy ; methods ; Finasteride ; therapeutic use ; Follow-Up Studies ; Hemospermia ; therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

ObjectiveTo detect the changes of gastroesophageal reflux disease (GERD) with high resolution endoscopy,and to explore its diagnostic value for nonerosive reflux disease (NERD). Methods From April 2007 to January 2008, consecutive out-patients visiting the Department of Gastroenterology due to continuous or recurrent symptoms of acid reflux, heartburn, cardiac-like chest pain for at least three months and volunteer healthy controls were recruited to the study. The subjects were classified into the normal group ( n =48 ), the NERD group ( n =70), the erosive esophagitis (EE) group ( n =70), and the Barrett esophagus (BE) group ( n =48). All subjects underwent endoscopy, and the shape of Z-lines, the shape of mucosal pits, mucosa roughness above Z-line and the shape of mucosal blood vessels were observed. The NERD changes were analyzed with optimal scale. ResultsZ-line shape of NERD group showed a significant difference from that of EE and BE groups ( P ＜ 0. 01 ), which was not different from that of normal controls ( P ＞0. 01 ). The shape of mucosal pits of NERD group was different from that of EE and BE (P ＜0. 01 ), which also was not different from that of control (P＞0. 01 ). Roughness of the mucosa above Z-line of NERD group was different from other 3 groups (P ＜0. 05). And the shape of mucosal blood vessels of NERD was different from the control (P＜0. 05), but was not from two others. Features of NERD relative images were thin,straight and spiral blood vessels, bar-like pits, plat mucosa, round, smooth and wide tooth-like Z-line and mucosa of white particle hypertrophy. ConclusionHigh resolution endoscopic features of most NERD patients are cloudy, white and rough mucosal surface, with white particular hypertrophy, spiral vascular dilation, extending to Z-line. These features can be indicators of NERD.

Objective To explore the changes of S-100 protein(S-100) levels in blood in newborn rats with bilirubin encephalopathy.Methods The model was established by administered bilirubin intraperitoneally(200 mg/kg) in newborn rats.The blood and brains samples were taken from 8 rats at the sequential time points of 6,12,24,48,72 and 96 h after the model established.Eight normal rats were used as the control.The dynamic changes of S-100 were detected by enzyme linked immunosorbent assay(ELISA) and immunohistochemical technique.Results S-100 significantly increased in blood of bilirubin encephalopathy newborn rats compared with controls(P

Arachidonic acid is an essential fatty acid in human nutrition and a biogenetic precursor of the biologically active prostaglandins and leukotrienes. ?~5 desaturase catalyzes the ?~5 dehydrogenation of di-homo-?-linolenic acid to form arachidonic acid in biosynthetic pathway of arachidonic acid. Using real time PCR technology,the transcriptional expression levels of gene encoding ?5 desaturase in three Mortierella alpina strains M10,M6 and M23,and in different growth phase of high arachidonic acid yielding strain M6,were determined. Results showed that there was a distinct corelationship between mRNA transcript level of ?~5 desaturase gene and biosynthesis of arachidonic acid. Results indicated that ?~5 desaturase plays an important role in arachidonic acid biosynthesis.

Aim To investigate estradiol inhibition of neointimal proliferation after rat carotid artery balloon injury. Methods Eight to ten-week-old SD rats (male,n=21,female,n=21) were divided into intact control(n=7),gonadectomy control(n=7) and estradiol (n=7, gonadectomy)groups in each sex. Left carotid artery was not injured with 2.0 F PTCA balloon until estradiol was injected for three days. Rats were killed 2 wk after injury. Neointimal areas and media area, ratios of intimal areas/media areas were measured with computer. Results Male neointimal areas and ratios of intimal areas /media areas in estradiol group were less than those in intact control group significantly(all P0.05). Conclusions Estrdiol inhibits neointimal proliferation after the gonadectomy in rats undergoing carotid artery balloon injury.

Fourteen compounds were isolated by column chromatography from the ethyl acetate extract of the seeds of Brassica campestris. Their structures were elucidated by physicochemical properties and spectroscopic data analysis. The isolated compounds were respectively identified as (5Z,7E)-4, 4-dimethyl-5-acetyl-5, 7-nonadienoic acid (1), indole-3-carboxaldehyde (2), blumenol A (3), vinylsyringol (4), sinapinic acid (5), sinapic acid ethyl ester (6), protocatechuic acid (7), crinosterol (8), campesterol (9), 7-oxo-stigmasterol (10), kaempferol (11), 2,5-dihydroxybenzoic acid (12), syringic acid (13) and daucosterol (14). Compound 1 was a new compound and the other compounds were isolated from this plant for the first time except for compounds 4, 5 and 13.
Brassica ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Seeds ; chemistry ; Spectrometry, Mass, Electrospray Ionization