Objective To investigate the MRI features of the brain in patients with neuromyelitis optica (NMO), and to evaluate the correlation between the brain abnormalities and related risk factors.Methods Fifty-four patients with definite NMO according to 2006 Wingerchuk diagnosis criteria were enrolled in this study. MRI scanning of the brain was performed in these patients. Distribution and signalfeatures of all the lesions were analyzed. A Logistic regression analysis was used to evaluate the risk factors of brain abnormalities. Results Twenty-four NMO patients (44. 4%) showed unremarkable findings and thirty (55.6%) showed abnormalities on brain MRI. Multiple and non-specific small lesions in the subcortical white matter and grey-white matter junction were the most frequent abnormalities on brain MRI (13/30, 43. 3%). Typical lesion locations included corpus callosum, subependyma of ventricles,hypothalamus and brain stem. The lesions showed punctate, patchy and linear abnormal signals. Postcontrast MRI showed no abnormal enhancement in 16 cases. Logistic regression analysis showed that coexisting anto-immune disease or infection history had correlations with abnormalities of the brain on MRI (OR=3.519,P ＜0.05). Conclusions There was a high incidence of brain abnormalities in NMO.Subependymal white matter, corpus callosum, hypothalamus and brain stem were often involved in NMO.NMO patients with coexisting anto-immune disease and infection history had higher risk of brain abnormalities.

Objective To investigate the effect of VvhA recombinant protein to the expression of IL-8 in human intestinal epithelial Caco-2 cells. Methods The VvhA recombinant protein(rVvhA) was ex-pressed by prokaryotic expression vector pET-28a (+)-whA in E. coli BL21 (DE3) , purified by Ni~(2+)-NTA affinity chromatography refoldod by stepwise deliquation together with dialysis methods and identified by Western blot. The cytotoxic of rVvhA to human Caco-2 cells was measured by CCK-8. The transcription of IL-8 mRNA in human Caco-2 cells induced by rVvhA was determined by RT-PCR, and the expression of IL-8 in human Caco-2 cells induced by rVvhA was determined by ELISA assay. Results rVvhA was purified with high purity up to 95%. The viability of human Caco-2 cells treated with 1.5 HU/ml rVvhA was inhibi-ted significantly (P < 0.05). The rVvhA can induce human Caco-2 cells to increase the transcription and expression of IL-8 in dose- and time-dependent manner. The transcription of IL-8 gene in human Caco-2 cells treated with 0.6 HU/ml rVvhA in 30 min can be up-regulated significantly, and the expression of IL-8 in human Caco-2 cells treated with rVvhA in 4 h can be increased significantly. Conclusion rVvhA has cy-totoxic to human Caco-2 and can increase the expression of IL-8, it might play a major role in the inflamma-tory reaction of rVvhA-exposed cells.

Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on hemolysin gene(vvhA)that coding cytolysin.Method By using software Primer Express, the PCR primers and TaqMan probe,which located in the conserved region of vvhA gene sequence,were designed for establishment of a TaqMan real-time fluorescent quantitative PCR to detect 100 bp amplicon from V.vulnificus DNA.A recombinant plasmid pMD19-vvhA100 as a positive control during detection was constructed using gene cloning technique.Minimal amplification cycles(Ct value)and fluorescence intensity enhancement (△Rn value)were used as observing index to optimize the reaction conditions of the TaqMan real-time fluorescent quantitative PCR.The DNAs with different concentrations from V.vulnificus and other eight bacteria and pMD19- vvhA100 were applied as templates to determine the specificity,sensitivity and reappearance of the TaqMan real- time fluorescent quantitative PCR.ICR mice were intraperitoneally,subcutaneously and orally infected with V. vulnificus,respectively.The detection effect of the TaqMan real-time fluorescent quantitative PCR was measured using the specimens of peripheral blood,subcutaneous tissue and intestinal content collected from the infected mice.Results The established TaqMan real-time fluorescent quantitative PCR showed positive results only for V. vulnificus DNA and pMD19-vvhA100.The detection effectiveness of the TaqMan real-time fluorescent quantitative PCR was as high as 0.01 ng of V.vulnificus DNA or 103 copies of pMD19-vvhA100.The SD values of the detection results repeated for three times using pMD19-vvhA100 with different concentrations were lease than 0.79. The detection results of TaqMan real-time fluorescent quantitative PCR were positive for all the specimens of peripheral blood and subcutaneous tissue.Conclusions The TaqMan real-time fluorescent quantitative PCR established in this study for V.vulnificus vvhA gene detection has advantages such as quickness,stability, sensitivity and specificity,indicating this method can be used for clinical laboratory diagnosis of septicemia and wound infection caused by V.vulnificus.

Background Endophthalmitis is a serious,sight-threatening condition.Identifying the causative microorganisms is very important for available treatment of endophthalmitis.Objective This survey was to analyze the spectrum of organisms causing culture-proven endophthalmitis and their sensitivities to commonly antimicrobial agents.Methods Medical data of patients with culture-proven endophthalmitis at Zhongshan Ophthalmic Center from January 2003 through December 2010 were respectively reviewed.The outcomes included intravitreal isolates and antibiotic sensitivities were analyzed.Results Four hundred and sixty-nine strains of organisms were isolated from 447 eyes of 447 patients with infective endophthalmitis,including 22 eyes of polymicrobial infection.In the organisms,gram-positive organisms were 241 (51.4％),fungi were 125 (26.7％) and gram-negative organisms were 103 (22.0％).The most common organisms were Staphylococcus epidermidis in 29.4％,Aspergillus in 7.7％ and Pseudomonas aeruginosa in 5.3％.In this group of infective patients,the most common clinic settings were posttraumatic endophthalmitis (72.7％),and then were postoperative endophthalmitis (10.5％),endogenous endophthalmitis (9.8％) and keratitis (6.9％).Most gram-positive organism and gram-negative organism were sensitive to levofloxacin and cefoperazone.The susceptibility rate of gram-positive organism to chloromycetin was increased in 2007-2010 years compared with 2003-2006 years (x2=5.398,P＜0.05).The susceptibility rate to ciprofloxacin of gram-negative organisms declined (x2 =5.398,P ＜ 0.05),but that to rifampicin increased in the duration of 2007-2010 compared with 2003-2006 (x2 =4.500,P ＜ 0.05).Conclusions Gram-positive organisms are the most commonly causative organisms of endophthalmitis.Most bacterial organisms are sensitive to levofloxacin and cefoperazone.Local data of culturing and susceptibility test offers a guideline for the treatment of infectious endophthalmitis.

OBJECTIVETo summarize and analyze the epidemic situation of human rabies from 1984 to 2002 in China, and to explore the possible factors causing the increase of cases so as to provide evidence for preventive and control measures.

METHODNational and some provincial data on the prevalence of rabies during 1984 to 2002 were collected and analyzed.

RESULTSFrom 1984 to 1989, the annual reported cases were between 4 000 and 6 000 but decreased after 1990. In 1996, the reported cases decreased to the lowest level from 3 520 in 1990 to 159. However, number of reported cases has been continuously increasing since 1998 which reached 1 122 in 2002, a 7.06 times increase as compared to the number in 1996. The epidemic areas were mainly located in the southeast and southwest parts of the country, such as Sichuan, Hunan, Guangxi, Guangdong, Anhui, Fujian, etc. Furthermore, there was no significant seasonal distribution as it showed before.

CONCLUSIONSuch facts as the increasing numbers of dogs, low inoculation rate to dogs, poor control on the quality of rabies vaccine, mistreatment to the wounds, and lacking good cooperation between different official departments regarding rabies control might serve as important factors responsible for the recurrence of rabies. Therefore, it is necessary to focus on the above mentioned points and to take comprehensive preventive measures to bring down the prevalence of rabies in China.

Animals ; China ; epidemiology ; Dogs ; Humans ; Rabies ; epidemiology ; prevention & control ; Rabies Vaccines ; standards ; Seasons ; Time Factors

OBJECTIVETo establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on the hemolysin gene (vvhA) coding cytolysin.

METHODSPrimers and probes in the conserved region of the vvhA gene sequence were designed for the TaqMan real-time PCR to detect 100 bp amplicon from V. vulnificus DNA. Recombinant plasmid pMD19-vvhA100 was constructed and used as a positive control during the detection. Minimal amplification cycles (Ct value) and fluorescence intensity enhancement (DeltaRn value) were used as observing indexes to optimize the reaction conditions of TaqMan real-time PCR. The TaqMan assay for the detection of Vbirio vulnificus was evaluated in pure culture, mice tissue which artificially contaminated Vibrio vulnificus and clinical samples.

RESULTSThe established TaqMan real-time PCR showed positive results only for Vibrio vulnificus DNA and pMD19-vvhA100. The standard curve was plotted and the minimum level of the vvhA target from the recombinant plasmid DNA was 10(3) copies with a Ct value of 37.94+/-0.19, as the equivalent of 0.01 ng purified genomic DNA of Vibrio vulnificus. The results detected by TaqMan PCR were positive for the 16 clinical samples and all the specimens of peripheral blood and subcutaneous tissue of mice which were infected with Vibrio vulnificus.

CONCLUSIONTaqMan real-time PCR is a rapid, effective, and quantitative tool to detect Vibro vulnificus, and can be used in clinical laboratory diagnosis of septicemia and wound infection caused by Vibrio vulnificus.

Animals ; Bacterial Proteins ; genetics ; Base Sequence ; DNA Primers ; Mice ; Mice, Inbred ICR ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Vibrio vulnificus ; isolation & purification

The aim of this study is to investigate the effect and mechanism of angiotensin (Ang) II on E-selectin and vascular cell adhesion molecule-1 (VCAM-1) expression in rat brain microvascular endothelial cells (BMEC) and evaluate the effect of compound EXP-2528, a novel Ang II type 1 (AT1) receptor antagonist. The experiment was performed in cultured BMEC of rat. The mRNA and protein expression of E-selectin and VCAM-1 in BMEC was analyzed by RT-PCR and Western blotting, respectively. The results showed that the mRNA and protein expression of E-selectin and VCAM-1 in BMEC were significantly upregulated by 4 h or 18 h exposure to 1 x 10(-7) mol x L(-1) Ang II. These effects were abolished by pretreatment with the selective AT1 receptor antagonists losartan and compound EXP-2528, but not with the AT2 selective antagonist PD123319. Combining losartan with PD123319 also significantly inhibited Ang II-induced E-selectin and VCAM-1 expression in BMEC, but there was no significant difference compared with losartan group. These findings indicated that Ang II upregulated E-selectin and VCAM-1 in BMEC by activating AT1 receptor and then involved in the development of cerebrovascular disease.
Angiotensin II ; pharmacology ; Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Brain ; blood supply ; Cells, Cultured ; E-Selectin ; genetics ; metabolism ; Endothelial Cells ; metabolism ; Imidazoles ; pharmacology ; Isoxazoles ; pharmacology ; Losartan ; pharmacology ; Microvessels ; cytology ; RNA, Messenger ; metabolism ; Rats ; Vascular Cell Adhesion Molecule-1 ; genetics ; metabolism

OBJECTIVETo explore the roles of matrix metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) in the pathogenesis of endometriosis and the effects of estrogen and progestin on their expression.

METHODSImmunohistochemistry and RT-PCR were employed to detect the expression of MMP-1 and TIMP-1 in the ectopic tissues of 35 patients with endometriosis, 22 eutopic endometrium tissues from women with endometriosis and 28 normal controls. Fifty-nine nude mice were injected with human late secretory endometrial chippings and randomized into estrogen group, progestin group, estrogen-progestin group and control group with corresponding treatments. The implantation rates and graft morphology were observed and MMP-1 and TIMP-1 expressions in the grafts detected by immunohistochemistry.

RESULTSTypical endometrial glands and stroma were observed in all the groups with comparable implantation rates. The administration of progestin was associated with multiple peritoneal implantation sites and significantly larger implants. The transplanted endometria showed proliferative or secretory changes with estrogen or progestin administration. MMP-1 expression significantly increased and TIMP-1 expression decreased with increased MMP-1/TIMP-1 ratio in human and nude mouse ectopic endometria in comparison with those in normal endometria (P<0.05, P<0.01). MMP-1 expression was higher in estrogen and estrogen-progestin groups than in the control group, and was lower in the 3 sexual hormone-treated groups than in the control group. MMP-1 mRNA expression in the eutopic endometrium was significantly higher than that in the normal endometria.

CONCLUSIONProgestrin can not inhibit MMP-1 expression or the effect of estrogen on ectopic endometrium known as progestin resistance. The high expression of MMP-1 and low expression of TIMP-1 in endometriotic tissues confer strong invasiveness of ectopic endometrial tissue, especially in eutopic endometrial tissue, and may play an important role in the pathogenesis of endometriosis.

Adult ; Animals ; Endometriosis ; metabolism ; Estrogens ; pharmacology ; Female ; Humans ; Matrix Metalloproteinase 1 ; genetics ; metabolism ; Mice ; Mice, Nude ; Middle Aged ; Progestins ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

OBJECTIVETo investigate the expression of microRNA-21(miR-21) in ovarian epithelial carcinoma and its association with the clinicopathological features.

METHODSThe expression of miR-21 was detected by Stem-loop real-time RT-PCR in 48 cases of ovarian epithelial carcinomas, 24 cases of benign ovarian epithelial tumors and 15 cases of normal ovarian tissues.

RESULTSThe relative expression level of miR-21(2-(DeltaDelta)CT) was 4.849-/+1.813 in the ovarian epithelial carcinomas, significantly higher than that in the benign ovarian tumors and normal ovarian tissues (P<0.01), but comparable between the latter two groups. The expression of miR-21 was not correlated to the histological type, but increased significantly with the progression of the clinical stages and histological grading (P<0.01), showing a close correlation to lymphatic metastasis.

CONCLUSIONMiR-21 might play a role as an oncogene in the tumorigenesis and development of ovarian epithelial carcinoma, and is possibly correlated to the progression and prognosis of ovarian epithelial carcinoma.

Adult ; Aged ; Cystadenocarcinoma, Serous ; genetics ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Ovarian Neoplasms ; genetics ; metabolism ; Prognosis ; Young Adult

Objective To investigate the ostengenie potential of adipose-derived stem cells(AD- SCs)when exposed to adenovirns containing hBMP-2 cDNA(Adv-hBMP-2)and offer a choice of cell source for gene therapy and tissue engineering.Methods Human adipose tissues were obtained from patients who received orthopaedic surgery or liposuction.ADSCs were obtained by digesting the adipose tissues.Firstly,flowcytometric analysis was performed for the confirmation of mesenchymal stem cell ori- gin and the surface markers including CD34,CD44,CD68,CD71,CD90,and CD105.The ADSCs were transfected by Adv-hBMP-2 and the effects were tested in vitro,lmmunoprecipitation and Western blotting and ELISA were performed for confirming BMP gone transduction and its stable expression.The transform of ADSCs was assessed by extracellular ALP staining,intracellular ALP spectrophotometry,von Kossa staining and RT-PCR.In the in vivo experiment ADSC-Adv-hBMP-2 cells were injected into the hind limb of nude mice and analyzed radiographically and histologically.Results ADSCs were successfully isolated from human adipose tissues.The isolated ADSCs expressed CD44,CD71,CD90 and CD105 and CD34 and CD68 were absent.The result confirmed the mesenchymal stem cell origin of the cells.West- ern blotting and ELISA confirmed successful and persistent hBMP-2 production by ADSC-Adv-hBMP-2 cells.Extracellular ALP staining,intracellular ALP spectrophotometry,yon Kossa staining and RT-PCR revealed that ADSCs treated with Adv-hBMP-2 had a tendency of transfering into osteoblast.X-ray and H&E sections from hind limb of nude mice injected with ADSC-Adv-hBMP-2 cells confirmed bone forma- tion at 2 weeks.Conclusions Liposuction aspirates contain abundant ADSCs that can be transduced with hBMP-2 gene,and the tranduced ADSCs differentiate into the osteoblast.ADSCs may be an ideal source of mesenchyme-lineage stem cells for gone therapy and tissue engineering.