The quorum sensing commonly exists in procaryote kingdom,regulating various biologic functions.Recently similar phenomenon was also found in fungi world,which affecting both biofilm formation and dimorphism.In this article,we focus on the recent progress on quorum sensing of pathogenic fungi and discuss the possibility of taking quorum sensing molecule as a potential target for antifungl therapy.

BACKGROUND: Simple drug therapy can not acquire satisfactory effects in treatment of fungal keretitis. At present, penetrating keratoplasy (PKP) has been considered a primary means to cure the fungal keratitis to save eyeball and vision.OBJECTIVE: To evaluate the clinical efficacy of PKP in the treatment of high-risk fungal keratitis.DESIGN, TIME AND SETTING: A retrospective case analysis was performed at the Department of Ophthalmology, Affiliated Hospital of Qingdao University Medical University between January 2000 and January 2007.PARTICIPANTS: A total of 51 patients (51 eyes) with high-risk fungal keratitis who underwent PKP in the Affiliated Hospital of Qingdao University Medical College were recruited into this study. Of these patients, 12 suffered from perfored, 35 from hypopyon, 8 from complicated cataract prior to surgery, and 5 from complicated glaucoma.METHODS: All patients received antifungal and antibacterial treatments prior to surgery and underwent PKP within 4 days following admission. After surgery, antifungal and antibacterial treatments were performed locally and systemically. All patients were followed-up for 6--24 months.MAIN OUTCOME MEASURES: Postoperative visual acuity, recurred fungal infection, rejection of implants, secondary glaucoma, and ulceration of implant.RESULTS: ① Of 51 patients, 18 were followed-up for 6-12 months, 2 for 13-18 months, and 8 for 19-24 months. ② A total of 49 (96.1%) out of 51 patients preserved the eyeballs and the visual acuity improved to different degrees in 48 (94.1%) patients. ③ After surgery, fungal infection recurred in 6 eyes (11.6%), 4 of which were controlled by antifungal medication and 2 was enucleated because of uncontrolled endophthalmitis. Graft rejection was found in 18 (35.3%) eyes, 13 of which recovered transparent by medication and 5 received secondary PKP. Graft ulceration was present in 4 (7.84%) eyes, 3 of which were cured and the remaining one was re-grafted because of severe endothelial cell loss. Secondary glaucoma appeared in 7(13.7%) eyes, and the intraocular pressure was controlled medically and surgically. Complicated cataract occurred in 6 (11.8%) eyes, 3 of which underwent cataract extraction. Most complications were successfully controlled. In the final follow-up period, 45 (88.2%) grafts were transparent.CONCLUSION: PKP is an effective approach to preservation of eyeballs and restoration of visual function in patients with high-risk fungal keratitis, which can not be treated by conservative therapy.

Objective To clarify the quorum sensing and identify the quorum sensing molecular of Cryptococcus neoformans. Methods The growth of Cryptococcus neoformans TUP1 △ strain at different inoculum size was observed. The culture filtrate of high cell density TUP1△ strain was collected and plated on medium with low density TUP1△ cell, and the growth of low cell density TUP1△ was recorded. Finally, the active molecule in culture filtrate was purified and identified.Results The growth of Cryptococcus neoformans TUP1△ strain was density dependent and the colony could be normally formed only above a specific density. The filtrate from a high cell density TUP1△culture could promote the growth of low cell density TUP1△, and the active molecule in this culture filtrate was identified to be an oligopeptide composed of 11 amino acids pepride, named quorum sensing peptide 1(QSP1). QSP1 was artificially synthesized, which could also promote the colony forming of TUP1△ strain inoculated at low cell density. Conclusion Quorum sensing exists in Cryptococcus neoformans, and the quorum sensing molecular is QSP1.

Fucoidan has been reported to exhibit various beneficial activities ranging from to antivirus and anticancer properties. However, little information is available about the effects of fucoidan on cerebral ischemia-reperfusion injury (IRI). Our study aimed to explore the effects of fucoidan on cerebral IRI, as well as the underlying mechanisms. Sprague-Dawley (SD) rats were randomly subjected to four groups: Sham, IRI+saline (IRI+S), IRI+80 mg/kg fucoidan (IRI+F80), and IRI+160 mg/kg fucoidan (IRI+F160). Fucoidan (80 mg/kg or 160 mg/kg) was intraperitoneally injected from 7 days before the rats were induced to cerebral IRI model with middle cerebral artery occlusion (MCAO) method. At 24 h after reperfusion, neurological deficits and the total infarct volume were determined. The levels of inflammation-associated cytokines (interleukin (IL)-1β, IL-6, myeloperoxidase (MPO), and tumor necrosis factor (TNF)-α), oxidative stress-related proteins (malondialdehyde (MDA) and superoxide dismutase (SOD)) in the ischemic brain were measured by enzyme-linked immunosorbent assay (ELISA). Besides, the levels of apoptosis-related proteins (p-53, Bax, and B-cell lymphoma (Bcl)-2) and mitogen-activated protein kinase (MAPK) pathway (phosphorylation-extracellular signal-regulated kinase (p-ERK), p-c-Jun N-terminal kinase (JNK), and p-p38) were measured. Results showed that administration of fucoidan significantly reduced the neurological deficits and infarct volume compared to the IRI+S group in a dose-dependent manner. Also, fucoidan statistically decreased the levels of inflammation-associated cytokines, and oxidative stress-related proteins, inhibited apoptosis, and suppressed the MAPK pathway. So, Fucoidan plays a protective role in cerebral IRI might be by inhibition of MAPK pathway.
Animals ; Apoptosis ; Brain ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Infarction, Middle Cerebral Artery ; Interleukin-6 ; Lymphoma, B-Cell ; Methods ; Peroxidase ; Phosphotransferases ; Protein Kinases ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; Reperfusion Injury* ; Superoxide Dismutase ; Tumor Necrosis Factor-alpha

Objective To investigate the value of intravoxel incoherent motion (IVIM) model of diffusion weighted MRI in assessing grades and enhancement patten of uterine cervical cancer. Methods Thirty one patients with pathologically proven cervical cancer, who underwent MRI scan preoperatively, were analyzed retrospectively and were divided into 3 groups according to their pathological grading of cacer, including 6 with G1 cancer, 17 with G2 and 8 with G3. The diameter of each lesion was≥1 cm. 10 b values (0, 30, 50, 100, 150, 200, 400, 800, 1 000, 1 500 s/mm2) were used in DWI, and DCE-MRI was performed with a time resolution of 9.8 s. Parameters of DWI (ADC, D, f, D*) and semiquantitative parameters of DCE-MRI (Slop, Maxslop, CER, Washout, AUC90) were measured. One-way ANOVA analysis of variance and Pearson correlation were used to analyze normally distributed continuous data. Kruskal-Wallis H test and Spearman correlation were used to analyze abnormally distributed continuous data. Tumor volume and all of the MRI parameters were compared as well as correlated with pathological grading.The perfusion parameters derived from IVIM were correlated with those derived from dynamic enhanced MR imaging. The sensitivity and specificity of f value to to diagnose G3 cervical cancer and the best cutoff were calculated from areas under the ROC curves.Results Tumor volume of G1,G2 and G3 cancers were(33.8±31.1),(19.6±16.9)and(31.2±29.1)cm3(F=1.147,P=0.332), respectively.ADC values of the three groups were(1.03 ± 0.11)× 10-3,(1.00 ± 0.10)× 10-3 and(0.90 ± 0.05)× 10-3mm2/s,respectively(F=4.619,P=0.018).D values of the three groups were (0.80 ± 0.11) × 10-3, (0.77 ± 0.06) × 10-3and (0.69 ± 0.06) × 10-3mm2/s ,respectively(F=5.272, P=0.011).f values of the three groups were 0.20±0.02, 0.22±0.03 and 0.24± 0.03, respectively (F=3.524, P=0.043).All of the others were of no significant difference (P>0.05).Both ADC and D correlated negatively with tumor grading (r=-0.464 and-0.493, P=0.009 and 0.005, respectively). f value correlated positively with tumor grading (r=0.436, P=0.014).Areas of ADC, D and f value under ROC curves to diagnose G3 cancers were 0.179, 0.147 and 0.690, respectively. While the cut-off value of f was 0.22, the diagnostic performance for G3 cancer was with a sensitivity of 75.0% (6/8) and a specificity of 60.9% (14/23). The value of f had weak positive correlations with Slop, Maxslop, CER and AUC90 of semiquantitative analysis of DCE-MRI (r=0.319, 0.337, 0.293 and 0.344, respectively, P<0.01). Conclusion IVIM model of multi-b value DWI may provide information in the assessment of differentiation and en hancement pattern of cervical cancer.

Objective To compare the detection efficiency between multiplex RT-PCR method and liquichip technology for screening the viral etiological agents of diarrhea.Methods The development of the multiplex RT-PCR method.A total of 107 feces samples from patients who suffered from diarrhea and attended to Zhujiang Hospital of Southern University from September 2013 to February 2014 were collected and tested in parallel by both multiplex RT-PCR and xTAG Gastrointestinal Pathogen Panel ( xTAG GPP) for Adenovirus, Norovirus genogroupⅠandⅡ, as well as by both multiplex RT-PCR and monoplex RT-PCR for Astrovirus and Sapovirus.To evaluate the sensitivity and specificity of multiplex RT-PCR, xTAG GPP and monoplex RT-PCR were used as reference.Kappa coefficient test was used to evaluate the consistency among the methods.The detection limit and accuracy of multiplex RT-PCR were evaluated by detection of serial dilution of positive plasmids and products sequencing for the five viral agents.Results The multiplex RT-PCR showed high consistency with xTAG GPP and monoplex RT-PCR, in which Kappa value was 0.885 and 1.000 respectively( P=0.000 ).Compared to xTAG GPP, the sensitivity and specificity of the multiplex RT-PCR were at average of 80.8%( 21/26 ) and 100%( 295/295 ) respectively.The detection limit and accuracy of multiplex RT-PCR were 104 copies /μl-106 copies/μl.Conclusion The high consistency indicated that both the multiplex RT-PCR and xTAG GPP are useful as a special,sensitive, high throughput and rapid diagnostic tools for the detection of the major viral pathogens related to diarrhea in clinical laboratory.

Objective: To evaluate the effect and safety of an anti-IL-8 monoclonal antibody cream for the treatment psoriasis vulgaris. Methods: Eight-four patients with psoriasis vulgaris were treated with the anti-IL-8 monoclonal antibody cream for 8 weeks,the safety and effect were evaluated by analyzing the adverse action and the changing of the diameter,color,thickness,scales of the lesions. Results: After the treatments for 8 weeks,11 patients were cured,41 patients were remarkably improved,the cure rate and effective rate is 13.1% and 61.9%,respectively.Among them the cure rate of guttate type and effective rate of mixed type were the highest,while the cure rate and effective rate for aggressive stage were both higher than those of stable stage.And only one patient had mild local adverse action. Conclusion: The anti-IL-8 monoclonal antibody cream was effective and safe for the treatment of psoriasis vulgaris.

Objective: To investigate the influence of TGF-β and IFN-γ on the proliferation, migration and invasion of melanoma cells. Methods: Melanoma cells were cultured in vitro. When tumor cells were confluent about 80% degree, cytokines were added into cell culture media. The concentration of TGF-β and IFN-β was 5ng/mL and 10ng/mL, respectively. Melanoma cells were divided into free-cytokine group, TGF-β group,IFN-γ group, TGF-β and IFN-γ group. Tumor cells in each group were then incubated for 8h, 16h, 24h, 32h,40h and 48h, respectively. After incubation, fixing and staining with SRB, the optical densities and percentage viability were then determined by absorption at 540 nm (A 540). The scarification of tumor cells in each group on the surface was created by a 2001μL pipette tube. The motility of tumor cells in each group was assessed by measuring the distance between scarifications. The speed of the scuffing closure was monitored after 12h.The invasive ability of melanoma cells was observed by transwell cultivation. The tumor cells that invaded through the Matrigel and adhered to the bottom of the outside membrane were determined by absorption at 595 nm (A595). Gelatin zymography assay was used to examine the levels of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) activity when the tumor cells were treated with cytokines after 24h. MMP-2 and MMP-9 activity was demonstrated by gradation in the sodium dodecyl sulfate polyacryl-amide gel electrophoresis (SDS-PAGE) gelatin. MMP-2 and MMP-9 activity was determined by Image analy-sis Software. Results: TGF-β promoted the proliferation, migration and invasion of melanoma cells (P<0.05).However, IFN-γ inhibited the proliferation, migration and invasion of melanoma cells (P<0.05). The effect weakened or disappeared when both of them were used (P>0.05). Conclusion: In vitro, TGF-β may affect the inhibitory effect of IFN-γ on the proliferation, migration and invasion of melanoma cells. This study provided a better understanding of the relationship between tumor and inflammatory factors and established a good ba-sis for future research.

Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.

Objective To observe the effect of small-interfering RNA(siRNA) targeting c-Met,the receptor of hepatocyte growth factor(HGF),on the proliferation of human lens epithelial cells(LECs).Methods siRNA was transferred into LECs cultured in vitro by HiperFect Transfection Reagent.Real-Time PCR was applied to observe the expression of c-Met mRNA in LECs after gene transfer,and MTT assay was used to detect the proliferation of LECs induced by HGF.Results The expression of c-Met mRNA in LECs was significantly decreased in the experimental group,compared to that in the controls(P<0.01).Proliferation of LECs induced by HGF was inhibited,compared with the single HGF stimulated group(P<0.01).Conclusion The RNA interference targeting c-Met can effectively inhibit the expression of c-Met mRNA,and the proliferation of LECs induced by HGF.