Objective: design knee prostheses infection modal for rabbits. Methods:The knee implants were designed with 3mm To Kirschner wire for the length of 15mm.Implanted the stem to the distal femoral through intercondylar fossa.forty eight rabbits were divided into 4 groups randomly,12 in each group and finished above-mentioned surgery. Immediately after the surgery,0.5 ml 1?107 CFU of Staphylococcus aureus ATCC 25923 was injected into the joints of group A,0.5 ml 1?106 CFU in group B and 0.5 ml 1?105 CFU in group C, group D were not injected.the animals were sacrificed 2 weeks after injections, wounds were observed and X-ray films were and taken. levels of serum CRP and ESR determined,pathological changes were observed and bacteria cultures were performed. Results: After staphylococcus aureus was injected into the joints right after the surgery.100% knee joint infection in group A and 100%in group B, 83.3% in group C,none knee joint infection in group D. A large amount of inflammatory cells were found in the infected samples, theand levels of serum CRP and ESR were elevated in the infected samples. Conclusion:An animal model of TKA infection can be established successfully at 2 weeks by 0.5 ml 1?106 CFU staphylococcus aureus ATCC25923 injecting into the joints immediately after the surgery.

BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)are widely utilized as seed cells or carriers in bone tissue engineering and gene therapy.Thus,how to obtain BMSCs with high purity arose more attentions of researchers.OBJECTIVE:To observe the effects of different separation methods and culture conditions on biological characteristics of rabbit BMSCs.METHODS:BMSCs were obtained by whole bone marrow culture,density grand centrifugal and red blood cell lysis.At 48 hours after culture,the cell numbers were counted,the time of passage was recorded,in addition,the cell morphology was observed by phase contrast microscope,and the CD44 antigen expression was identified using flow cytometry.The 3rd and the 7th generation aging cells were cultured with DMEM-LG,MEM-HG,and DMEM/F12 culture medium.MTT and count cell plat were used to evaluate the growth of BMSCs.Phase contrast microscope was used to observe the morphological changes of aging cells.RESULTS AND CONCLUSION:BMSCs could be separated by each method.The adherent cells showed shuttle or multiple angle shapes,with rich cytoplasm,and positive for CD44 antigen.The more cell number and shortest primary culture time was presented in red blood cell lysis group(P ＜ 0.05).DMEM/F12 could promote the proliferation of quiescent cells.And the cells prevented the better viability.The method of read blood cell lysis improving the efficiency of BMSCs adherent is an effective method of extraction of BMSCs.DMEM/F12 could promote the proliferation Maybe,DMEM/F12 is more suitable for BMSCs.

The general theories of gray correction methods for the medical ultrasonic image are introduced in this paper, including gray level correction, gray level transformation and histogram correction. The dominant histogram equalization method is expatiated and modified against its disadvantages including inapparent enhancement of partial detail and non-interactivity. Series connections of several gray correction methods are used to treat with the ultrasonic image, and thus its utility is improved through the selective enhancement of the contrast of certain gray scope.

Medical image segmentation is a classical puzzle for researchers.Image segmentation is the method to delineate anatomic structures or other interested regions automatically or semi-automatically,which is helpful to diagnosis and plays a crucial role in many medical imaging applications.In this article,a rather complete survey to medical image segmentation methods and their characteristics are given,especially to the new methods or improvement of the classical methods which was put forward recently.The prospects and the challenge of medical image segmentation is discussed.

Objective To determine the influence of P-glycoprotein (P-gp) inhibitor on the blood brain barrier (BBB) transport of amphotericin B (AmB)..Methods An in-vitro BBB model was established with brain capillary endothelia cells (BCEC). AmB was chosen as the test drug and verapamil was chosen as the inhibitor of P-gp.Cellular uptake of AmB at different time points and with series of verapamil concentrations were performed respectively after the determination of appropriate incubation time and drug dosage by the cytotoxicity assay. The AmB concentrations of series of samples were detected using high performance liquid chromatography (HPLC) method. One-way ANOVA analysis and Bonferroni test were used for data analysis.Results The cellular transport of AmB was accumulated as the time prolonged.The inhibitor group had a significant higher cellular uptake levelsof AmBat the time point of 90 min (t=6.753,P=0.001),120 min (t=3.574,P=0.016) and 150 min (t=4.759,P=0.005) as compared with the control group.The AmB cellular uptake level increased significantly when BCEC were incubated with verapamil of 2 μmol/L (P=0.000),5 μmol/L (P=0.014),10 μmol/L (P=0.000),50 μmol/L (P=0.014),75 μmol/L (P=0.000) and 100 tμmol/L (P=0.000),respectively,compared with the control group.Conclusion The P-gp inhibitor verapamil can enhance the cellular uptake of AmB which indicates that P-gp is involved in the BBP transport of AmB.

Objective To understand the clinical epidemiology of nosocomial candidemia in Huashan Hospital during a 10-year period. Methods One hundred and nine cases of nosocomial candidemia in Huashan Hospital affiliated Fudan University during the period of 1998- 2007 were retrospectively reviewed. The underlying conditions, risk factors, clinical manifestations, treatment and outcome were described. The prognostic factors were analyzed by chi square test or Fisher exact probability test. Multivariate analysis was done by multiple Logistic regression. Results The average annual incidence of nosocomial candidemia during the study period was 0.28/10 000 patients per day.The most common pathogen was C. albicans (59/109,54.1%), followed by C. tropicalis (20/109,18.3%), then C. parapsilosis (11/109, 10. 1%), C. glabrata (11/109, 10.1%), and other Candida spp. (8/109, 7.3% ). Underlying diseases frequently identified included diabetes (50,45.9%), solid malignancy (32, 29.4%), head trauma (13, 11. 9%) and stroke (12, 11.0%).There were 37 cases who died or deteriorated. The overall mortality was 34.0% and the attributable mortality was 22. 0% (24/109). In multivariate prognostic analysis, retention of central venous catheters (OR: 5.42, 95% CI: 1.68-17.41, P＝0.005), corticosteroid medication (OR: 3.69,95% CI: 1.10-12.34, P=0. 034), and severe sepsis on the day of candidemia (OR: 2.94, 95% CI:1.72-15. 21, P = 0. 003) were factors independently correlated to increased mortality. Furthermore,adequate antifungal therapy was the only independent predictor of decreased overall mortality (OR: 0. 27,95% CI: 0. 09-0. 78,P=0.015). Conclusions The incidence of nosocomial candidemia in our hospital has been increasing during the past decade. Timely diagnosis and treatment plays a key role in the management of nosocomial candidemia,

Surface plasmon resonance (SPR) biosensors have become an advanced method for measuring biological molecular interaction. This article is focused on the principle and advantages of SPR biosensor chip technology, and its new applications in biomedical research fields. Its application prospects are also discussed.
Clinical Laboratory Techniques ; Drug Evaluation, Preclinical ; methods ; Surface Plasmon Resonance ; instrumentation

Objective To describe the distributions of FCGR polymorphisms in human immunodeficiency virus (HIV)-uninfected patients with cryptococcosis,and to investigate the association of FCGR polymorphisms with the susceptibility to cryptococcosis.Methods The distributions of the four functional polymorphisms,including FCGR2A 131H/R,FCGR3A 158F/V,FCGR3B NA1/NA2,and FCGR2B 232I/T were compared between 198 cryptococcosis patients and 190 healthy controls.The polymorphisms distribution patterns were also compared between patients with central nervous system (CNS) infection and those without CNS infection.Genotyping of eight single nucleotide polymorphism (SNP) in FCGR were performed by multiplex SNaPshot technology using DNA extracted from blood samples.The comparison between patients and controls was performed by chi square test or Fisher exact test.Results Compared to healthy controls,the frequency of FCGR2B 232I/I increased (65％ vs 53％,x2 =4.27,P=0.039,OR=1.652,95％CI:1.02-2.67) and that of FCGR2B 232I/T decreased (27％ vs 40％,x2 =5.77,P=0.016.OR=0.542,95％CI:0.33-0.90) in patients with cryptococcal meningitis.Among immunocompetent patients,the frequency of FCGR2B 232I/I was also over-presented (69％ vs 53％,x2=4.53,P =0.033,OR=1.958,95％CI:1.05-3.66) and the FCGR2B 232I/T genotype was also less frequently observed (24％ vs 40％,x2=5.14,P=0.023,OR=0.467,95％CI:0.24-0.91) compared to healthy controls.There were 117 cases with CNS infection and 81 non-CNS infection cases.The genotype of FCGR2A 131R/Rwas over-presented (19％ vs 6％,x2 =6.48,P=0.011,OR=3.52,95％CI:1.27-9.73) and the FCGR2B 232I/T genotype was under-presented (27 ％ vs 46 ％,x2 =7.56,P =0.006,OR=0.431,95％CI:0.24-0.79) in patients with CNS infection compared with those without CNS infection.Furthermore,the frequency of FCGR2B 232I/I genotypes increased (69％ vs47％,x2 =5.47,P=0.019,OR=2.479,95％CI:1.15-5.34) and the frequency of FCGR2B 232I/T decreased (24％ vs 51％,x2 =8.66,P=0.003,OR=0.307,95％CI:0.14-0.68) in immunocompetent patients with CNS infection compared with those without CNS infection.Conclusions FCGR2A 131H/R and FCGR2B 232I/T are associated with the susceptibility to cryptococcal CNS infection,which suggests that FcγRⅡA and FcγRⅡB may contribute to the pathogenesis of cryptococcosis.

Objective To analyze the clinical features of patients with uncommon fungal infections in central nervous system (CNS).Methods Thirty-five patients with uncommon CNS fungal infections who were admitted to Huashan Hospital from 1997 to 2010 were retrospectively reviewed.The pathogens,symptoms and signs.treatments of patients were evaluated.The data were analyzed by rank sum test and Fisher'S exact test.Results Twenty-nine of the 35 patients met the definition criteria of prover CNS fungal infections,while the other 6 had probable diagnosis.Predisposing factors were found in 86% of all patients.The most common pathogens were Aspergillus and Candida species.The symptoms and signs commonly occurred including fever(22 cases),headache(19 cases), cranial neuropathy(12 cases),and meningeal irritation sign(12 cases).High white blood cell count,high protein level,and low glucose level were the main findings of cerebrospinal fluid (CSF) analysis.Patients with cerebral aspergillosis were more frequently accompanied with immunocompromised conditions, and they often got CNS aspergillosis from hematogenous dissemination or direct extension of paranasal sinus infection.Cerebral granuloma and abscess were the common clinical characteristics of CNS aspergillosis.Cerebral candidiasis often arose from neurosurgical surgery or traumatic brain injury,and these patients were usually presented with meningitis.All patients were treated with antifungal drugs and (or) surgical intervention and 77%(27/35) of the patients achieved complete or partial responses. Antifungal agents combined with surgical resection might improve outcome of patients with CNS aspergillosis; while removal or replacement of drainage tubes in combination with antifungal treatment showed satisfactory efficacy in patients with cerebral candidiasis who usually had shunt manipulation. Conclusions The incidence of CNS fungal infection, such as cerebral aspergillosis and candidiasis, is increasing. Early diagnose and therapeutic intervention are crucial for improving outcome.

Objective To describe the distribution of mannose binding lectin (MBL) genetic polymorphisms in non-acquired immunodeficiency syndrome (AIDS) patients with cryptococcosis in China and to verify the association of MBL polymorphisms with susceptibility to cryptococcosis.Methods The case-controlled genetic association study was conducted and 167 non-AIDS patients with cryptococcosis and 208 healthy controls were recruited. Genome DNA was extracted from the peripheral blood and MBL gene was amplified by polymerase chain reaction (PCR). Six singlenucleotide polymorphisms ( SNP) of MBL gene were sequenced. The association of MBL polymorphisms with susceptibility to cryptococcosis were analyzed. The comparison between patients and controls was performed by chi square test or Fisher's exact test. The differences of MBL plasma concentrations between groups with different MBL genotypes were compared by single factor variance analysis. Results There were no differences between patients and controls in terms of MBL genotype frequencies, haplotypes and genotypes (all P>0. 05). Compared with healthy control, the deficient MBL-producing genotypes were strongly associated with cryptococcal meningitis (16. 5% vs 8. 7%,χ2=4.25, P=0.0392, OR = 2.09), particularly in patients without underlying immunocompromised conditions (21. 4% vs 8. 7%, χ2 =7. 15, P = 0. 0075, OR = 2. 88). Individuals with MBL deficiency genotypes showed significantly higher rates of central nervous system (CNS) cryptococcal infection rather than non-CNS cryptococcosis (16. 5% vs 3. 1%, Fisher's exact test, P = 0. 010, OR = 6. 13).The difference was even more significant in the immunocompetent patients (21. 4% vs 4. 0%, P =0.009, OR= 6. 55). Conclusion MBL deficiency is associated with cryptococcal meningitis and may play a role in CNS Cryptococcus infection.