Objective To perform comparative proteomic analysis of human ovarian cancer cell lines for detecting platinum-resistance associated proteins.Methods The total proteins of two sensitive (SKOV3 and A2780)and four resistant(SKOV3/CDDP,SKOV3/CBP,A2780/CDDP and A2780/CBP) human ovarian cancer cell lines were separated by two-dimensional gel electrophoresis(2-DE).The differentially expressed proteins were analyzed using image analysis software,stained with Coomassie Brilliant Blue,then identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and database searching.The mRNA and protein levels of the differentially expressed protein which was most significant in all of the four resistant cell lines were validated by RT-PCR and western blotting,respectively.Results Five proteins were found to be significant in four cell lines. Annexin A3 and destrin were up-regulated and nicotinamide-adenine dinucleotide phosphate(NADP)- dependent isocitrate dehydrogenase 1 was down-regulated in all the four resistant samples.Glutathione transferase omega 1 had an increased expression in the other three resistant cell lines except for SKOV3/CBP in which its expression was not changed.However,cofilin 1 represented a different trend.In the two resistant sublines of SKOV3,eofilin 1 had a down-regulation,but it had an up-regulation in the cell lines induced from SKOV3.The expression of annexin A3 was up-regulated by 3-20 fold and the results of RT- PCR and western blotting showed complete consistency with that by 2-DE.Conclusions Proteomic techniques are useful to the identification of the resistance-associated proteins in ovarian cancer platinum- resistant cell lines and five candidates have been found.The five differential proteins might become hopeful candidate biomarkers for resistance.

Objective Oligosaccharides of Rehmannia glutinosa were separated and purified by different molecular weight cut-off ultrafiltration(UF) membranes,and then ultrafiltrate was concentrated and purified by nanofiltration(NF) membrane.Methods Different molecular weight cut-off UF unit first and then NF unit were used in process.Results Oligosaccharides were separated by twice UF,the optimum separation conditions:the concentration of feed solution was 13—132 mg/mL,the operation pressure was 0.25—0.275 MPa,and the temperature was 20—40 ℃.Then ultrafiltrate was concentrated and purified by NF membrane,the optimum separation conditions:the operation pressure was 0.59—0.79 MPa,the temperature was 20—40 ℃,and the concentration multiplegot to three.Under this condition,the total extraction rate of oligosaccharide products was 46.63% and the purity was above 93.3%,the molecular weights of oligosaccharide products determined by gel filtration were less than 6 000.Conclusion The technology is not only simple and feasible but also easy to separate and purify the oligosaccharides of R.glutinosa effectively.

Objective To explore the MRI features of ejaculatory duct obstruction.Methods During January 2003 to Dccember 2010,transrectal ultrasonography (TRUS) was performed for 106 patients and underwent surgical treatment for ejaculatory duct obstruction.Among them,16 patients underwent MRI examination.The MRI features of ejaculatory duct obstruction in these patients were summarized.Results Ejaculatory duct cysts,ranging in size from 4 mm ×4 mm ×7 mm to 4 mm ×4 mm ×9 mm and locating in the paramedian line,were detected in 5 of the 16 patients; ejaculatory duct dilation located in the paramedian line was detected in 7 patients,with the internal diameter of 5 to 30 mm. After contrast injection,significant enhancement of the wall of the ejaculatory duct was observed in 2 patients.Mullerian duct cysts complicated with dilated ejaculatory duct and seminal vesicles were detected in 4 patients,in whom the cysts were located in the median line,ranging in size from 4 mm × 5 mm × 6 mm to 34 mm×35 mm ×44 mm,with inverted teardrop shaped pointing toward the seminal colliculus.ConclusionThe most common MRI features of ejaculatory duct obstruction are ejaculatory duct dilation and ejaculatory duct cysts.

Drugs, Chinese Herbal ; chemistry ; Drugs, Investigational ; Humans ; Plasma

Objective To explore the feasibility,efficiency and effect of wide-type p53 gene transfected into cervical cancer HeLa cells by ultrasound microbubble intensifier. Methods Based on the optimum parameter of ultrasound wave irradiation (0.5 W/cm2 ,30 s) selected in pre-experiment,HeLa cells were divided into 5 groups:(A)simple plasmid group,(B)plasmid-ultrasound group,(C)plasmid-ultrasound-microbubble group,(D)plasmid-liposome group,(E)blank control group. At 24-48h after transfection,the transfection efficiency was detected by fluorescence microscope,the mRNA expression of p53 was detected by RT-PCR,the cell cycle was detected by flow cytometry,and the inhibition rate of cells growth was detected by MTT assay. Results Fluorescence microscopy showed that the expression of green fluorescent protein in group C was higher than that in group Band group D (P

Objective To investigate the effect of the cerebral protection and possible mechanism of fasudil for hypoxic-ischemic cerebral damage (HIBD) in neonatal rats. Methods The HBID model was established, then the mice were randomly divided into different groups. The expressions ofα-SMA and ROCK-2 were detected in the newborn rats with ischemia. Results Compared with the model group, expressions of α-SMA, ROCK-2 decreased in each treatment group with significant differences (P < 0.05 or P < 0.01). Following with the increases of administration dose and the administration time, expressions of α-SMA, ROCK-2 decreased gradually with significant differences (P<0.05 or P<0.01). Conclusion Fasudil can reduce the expressions of α-SMA, ROCK-2 in the newborn mice with hypoxic-ischemic brain damage to attenuate the brain tissue hypoxic-ischemic injury. The protective effect on brain is significant by giving high-dose fasudil in the early neonatal rat HIBD (0 h).

Aim: Clone and characterize of the 5′- flanking region of the nitrate reductase (NR) gene derived from Dunaliella salina(D. salina). Methods : The genomic DNA from D. salina was respectively digested with BamHI, EcoRI, HindIII, Pst I, Sal I and Xba I. A genomic walking cassette was ligated to the ends of the digested DNA fragments, and then genomic walking libraries comprising BL, EL, HL, PL, SL and XL were constsucted. The 5′- flanking region of the NR gene from genomic walking libraries of D. salina was amplified by LA-PCR. The DNA sequences were analyzed with the software - Promoter Predictions. Isolated 5′-flanking regions fused to the GUS gene were tested for transient expression in the alga. Results: A single specific PCR product of about 1200bp in length from the HL library was generated. Also, several conserved motifs, such as CAAT-box, GAGA-box were found, which are related to regulation of transcription, and the putative binding sites of transcriptional factors such as EBP, EFII, NF-E1 and LV. BLAST showed that the DNA sequences shared high homology with 5′-upstream region of the NR gene from Dunaliella viridis. The isolated 5′-flanking regions were able to strongly drive GUS reporter gene expression, suggesting that it contains the promoter elements necessary for the transcription of the NR gene. The expression pattern of the GUS gene and the NR gene were similar, both ware induced by nitrate and repressed by ammonium. Conclusion: The cloned 5′- flanking sequences of NR gene derived from D. salina might be a specific promoter with the ability to“switch on or off” an expression of the heterologous gene in transgenic D. salina.

Standardized residency training is a systematic, standardized and homogeneous project. With the continuously increasing requirements for fine management, the management of standardized residency training is facing enormous challenges, and the construction of management information system is imperative. This paper introduces the basic modules design and application experience of management information system for standardized residency training in training hospitals, aiming at improving work efficiency, standardizing process management and ensuring training quality.

Objective To investigate the suppression of mrp1 and MRP1 induced by small interfering RNA and the restoration of sensitivity to chemotherapeutic drugs in the multidrug-resistant hepatocellular carcinoma cell lines HepG2/mrp1. Methods mrp1-targeted small interfering RNA duplexes were designed and composed and introduced into multidrug-resistant hepatocellular carcinoma cell lines HepG2/mrp1. The suppression of mrp1 mRNA and its gene product MRP1 was examined by RT-PCR and flow cytometry (FCM), respectively. MTT assay was performed to measure the reverse effect of small interfering RNA based on the results of ICs0. Results The overexpression of mrp1 mRNA and MRP1 was effectively suppressed by small interfering RNAs. The level of mrp1 mRNA in the transfected HepG2/mrp1 cells was reduced to (86.36±2.76)% and MRP1 to (89.38±3.76)%compared with those of the controls. The resistance to ADR was reversed five-fold, which indicated the restoration of sensitivity to drugs. Conclusion Small interfering RNA can inhibit mrp1 expression effectively and reverse the multidrug resistance mediated by MRP1.

Object To study the protective effect of Rodobryum roseum (Hedw) Limpr. on acute myocardial ischemia. Methods Rat acute myocardial ischemia models were prepared by ligating their left coronary arteries. Changes of their S-T segment was observed on lead Ⅱ dynamic ECG. Degree of ischemia was assessed by the extent of S-T segment elevation. Lactic dehydrogenase (LDH), creative phosphokinase (CPK), superoxide dismutase (SOD) and malondialdehyde (MDA) were determined at the same time. Results R. roseum can significantly alleviate S-T segment elevation, being most evident at the dose of 2 g/kg (P