Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

BackgroundMouse has been used in laboratory studies as the model of ocular diseases.Electroretinogram (ERG)is a non-invasive method for primary examination to evaluate retinal function.Though flash ERG has been widely applied in the mouse ocular disease model for the functional assessment of the retinal outer layer,pattern ERG(PERG) is seldom used for inner retinal evaluation.ObjeetiveThe present experimental study was to investigate the recording parameters and method,wave characteristics of PERG and influencing factors in mouse,and to build the foundation for further research.Methods Thirty C57BL/6 mice aged 6 weeks old were included in this research.RETLport ( Roland Consult,Germany) was adopted for the recording of PERG.The positive needle electrode was placed in the cornea,and the reference and earth electrodes were placed under the derma in the cheek and tail.The PERG under different temporal frequencies (0.5,1.0,2.0 and 4.0 Hz),and special frequencies (0.05,0.10,0.20 cpd) were recorded in a photopic environment,and different contrast ratio (95％ and 99％ ) of stimulator or different transmission bands ( 1-100 Hz,5-30 Hz) in the same temporal frequency and spatial frequency were regulated to analyze the influence on mouse PERG.The use of animals was in compliance to the Regulations for the Administration of Affairs Concerning Experimental Animals by the State Science and Technology Commission.ResultsThe latency of N1 PERG showed a negative N1wave at around 37 ms and positive P wave at about 86 ms in adult mice.The amplitude of N1-P was 2-6 μV.Different spatial frequency,temporal frequency and contrast can affect the final results,and the different temporal frequencies were statistically significant.The wave was stable and the amplitude was unaffected at 5-30 Hz transmission bands with pronounced interference (mean amplitude of N1-P waves were(3.40±0.71),(5.08±0.88),(3.21±1.54),(3.85±1.96)μV in 0.5,1.0,2.0,4.0 Hz,F=7.43,P=0.00).ConclusionsPERG wave from adult mouse is similar to that from human.It is a useful method in evaluating the inner retinal function.Appropriate stimulating parameters are critical for recording.

Objective To evaluate 18F-fluorodeoxyglucose (FDG) PET/CT in detecting occult malignancy in patients with suspected paraneoplastic neurological syndrome (PNS).Methods Twenty consecutive patients who underwent PET/CT scanning with the indication of suspected PNS were retrospectively reviewed.The gold standard of PNS was either cytology or clinical follow-up, and the final diagnosis was compared with PET/CT findings.Results Of the 20 patients, six were PNS.PET/CT detected nine cases.Six were true positive and three were false positive.The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of PET/CT were 100% (6/6), 78.57% (11/14), 85.00% (17/20),66.7% (6/9) and 100.00% ( 11/11 ) respectively.The treatment plan was modified based on the PET/CT results in 4 patients.Conclusions 18F-FDG PET/CT may play a role in detecting the underlying malignancy of PNS.It is also valuable in staging of the malignancy thus providing information for therapy decision making.

Objective To investigate the renal angiographic manifestations of severe hemorrhage following minimally invasive pereutaneous nephrostolithotomy (MPCNL), and to evaluate the technique of super-selective renal arterial embolization in treating the condition. Methods Forty-eight cases of severe hemorrhage following MPCNL treated with super selective renal arterial embolization in our department were retrospectively reviewed. The angiographic findings, results and complications of embolization procedures were analyzed. Results Two cases were of acute hemorrhage immediately after MPCNL, and the other 46 cases were of delayed hemorrhage 2 to 7 days after MPCNL. Of these 48 cases, 25 (52.1%) showed simple pseudo-aneurysms, 6 (12.5%) pseudo-aneurysms accompanied with arterial-venous shunts, 1 (2.1%) pseudo-aneurysm with extravasated contrast medium, 11 (22.9%) arterial-venous fistulas, 2 (4.2%) extravasated contrast medium from arterial branches, 1 (2.1%) renal capsular branches varix, 2 (4.2%) no lesion detected. Successful super-selective embolization was achieved in all 46 positive cases, and renal hemorrhage was stanched consequently. Polyvinyl alcohol foam embolization particles (PVA), gelfoam and coils were used in the procedures (PVA in 18 procedures, PVA +coil in 5, gelfoam in 10, geffoam + coil in 11, PVA + gelfoam + coil in 2). Post-embolization syndrome of various degrees were seen in all treated patients. A slight rise in blood creatinine levels was observed in 12 cases. Conclusion Super selective renal arterial angiography and embolization is the treatment of choice in patients who suffered severe hemorrhage due to MPCNL.

Calcinosis ; complications ; Decalcification Technique ; Humans ; Meningeal Neoplasms ; complications ; Meningioma ; complications ; Specimen Handling ; methods ; Ultrasonics

OBJECTIVETo evaluate the inhibitory effect of genistin combined with anastrozole on the growth and apoptosis of breast tumor tissue, and to study their anti-cancer mechanism by using the model of 7,12-dimethylbenz [alpha] anthracene (DMBA)-induced mammary tumors following ovariectomy in Sprague-Dawley (SD) rats.

METHODSThe DMBA induced postmenopausal SD rats were randomly divided into the control group, the genistein group, the anastrozole group, and the genistein combined with anastrozole group. The growth of tumors was observed in each group. The proliferation index and apoptosis index of tumor cells were determined. Moreover, estradiol (E2) and 17beta-HSD1 mRNA levels were determined by ELISA and RT-PCR respectively.

RESULTSThe tumor growth was inhibited in the genistein group and the anastrozole group. The inhibitory ratio was significantly higher in the genistein combined with anastrozole group (P < 0.05). Compared with the control group, levels of E2 and 17beta-HSD1 mRNA decreased more significantly in the genistein combined with anastrozole group (P < 0.05).

CONCLUSIONSGenistein could suppress the growth of mammary tumors in postmenopausal rats. It showed synergistic effect when combined with anastrozole, which resulted in reduced levels of E2 and 17beta-HSD1 mRNA. It had inhibitory effect on the growth of breast tumors.

17-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estradiol ; metabolism ; Female ; Genistein ; administration & dosage ; pharmacology ; Mammary Neoplasms, Experimental ; chemically induced ; pathology ; Nitriles ; administration & dosage ; pharmacology ; Ovariectomy ; Postmenopause ; Rats ; Rats, Sprague-Dawley ; Triazoles ; administration & dosage ; pharmacology

Objective To investigate the relationship between carotid intima media thickness (IMT) and pulse pressure index (PPI) in elderly hypertensive patients.PPI was defined as 24 h mean pulse pressure(PP)/24 h mean SBP.Methods One hundred and three elderly hypertensive patients were categorized by PPI level:group A (PPI

The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS 115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.

Objective To detect the histological characteristics of collagen nodules from hypertrophic scars (HS) and investigate the origin of collagen nodules.Methods The scar tissues were collected from patients with plastic operation.Morphological characteristics of collagen nodules were observed by light microscopy of HE-stained sections; expressions of type Ⅰ /Ⅲ collagens were observed by polarized light microscopy of sirius red-stained sections; expression and distribution of myofibroblasts (MFb)-specific protein (α-smooth muscle actin,α-SMA) were observed by immunostaining in order to observe level of local tissue tension.Results Collagen nodules varied in shape,not only sphereshaped,and in size.Moreover,abundant fibroblasts (Fb) with large and light-stained nuclei were seen in the nodules compared to non-nodule area,indicating that the cells located in the modules were active.Some collagen nodules were composed largely of collagen type Ⅲ (green),but some mainly contained collagen type Ⅰ (red or yellow),indicating the difference in the time of nodule formation.α-SMA was expressed mainly in the deep dermis equivalent to the level of reticular layer of the new scar tissues (2 months after injury) ; α-SMA was expressed mainly in the nodules of the old scar tissues (2-10 years after injury),but almost not in non-nodule areas except for a strongly positive staining in the vessels.Moreover,α-SMA presented a heterogeneous distribution in the collagen nodules,with stronger expression in the epidermal end than in the subcutaneous tissue end and stronger expression in the superficial dermis than that in the deeper part.It was suggested that there existed massive amount of MFb and high tension in the nodules arid that the tension distribution was not uniform in or between the nodules.Conclusions Collagen nodules are of varying shape and size and collagen types are associated with the time of nodule formation.Moreover,Formation of the collagen nodules may be closely related to the distribution and evolution of the local tissue tension.

AlM:To discuss the protective effect ofα-mangostin on retinal light damage in mice.METHODS:Totally 30 Balb/c mice, aged 6~8wk, were randomly divided into the control group, light-exposure group and α-mangostin group. Every group contained 10 mice. Mice of α-mangostin group were treated with alpha-mangostin at the dose of 30mg/( kg · d ) body weight by intragastric administration daily for 7d, and then exposed to white light at the 5th d. The light-exposure group and α-mangostin group were exposed to 5 000 ± 200lx white light-emmiting diodes (LEDs) for continuously 1h to establish the mice model of retinal light damage. Flash -electroretinograme was recorded 72h after light exposure. The changes in retinal morphology of mice were observed by light microscopy. Retinas were extracted to detect the malondialdhyde ( MDA ) content change of the retinal homogenate.RESULTS: Flash-electroretinogram ( F-ERG ) showed that retinal dysfunction was less severe in α-mangostin group than in light-exposure group ( P<0. 05 ). Light microscopy test showed that retina structural damage was less severe in α-mangostin group than in light-exposure group (P<0. 05). The level of MDA in retinal tissue of α-mangostin group was significantly lower when compared with light-exposure group (P<0. 05).CONCLUSlON: α-mangostin inhibits lipid peroxidation induced by light damage and protect retina against light damage.