Objective To explore the relationship between antifungal resistance and expression of a virulence factor phospholipase B1 in Candida albicans. Methods Total RNA was extracted from 15 fluconazole-resistant and 15 fluconazole-sensitive Candida albicans isolates. RT-PCR was performed to detect the expression of phospholipase B1 mRNA. Extracellular and intracellular proteins secreted by Candida albicans cells were concentrated using salting-out method and extracted by P0013B RIPA lysis buffer, respectively followed by Western-blotting analysis to detect the expression of extracellular and intracellular phospholipase B1 protein. Results The relative expression of phospholipase B1 mRNA was 0.6173 ± 0.1090 in fluconazoleresistant Candida albicans isolates, significantly higher than in fluconazole-sensitive isolates (0.2653 ± 0.0935,P ＜ 0.05). Increased expressions of extracellular and intracellular phospholipase B1 protein were noted in fluconazole-resistant Candida albicans isolates compared with fluconazole-sensitive isolates (0.4145 ± 0.2773 vs 0.2720 ± 0.2194, 0.1825 ± 0.1831 vs 0.2653 ± 0.0935, t = 2.703, 3.443, both P ＜ 0.05). Conclusions The expressions of phospholipase B1 mRNA and protein are elevated in fluconazole-resistant Candida albicans isolates, suggesting that the expression of phospholipase B1 may be associated with the occurrence of antifungal resistance in Candida albicans.

ObjectiveTo test the susceptibilities of Candida albicans to 5 antifungal agents including methylrosanilinium chloride and to estimate their relationship with the genotypes of C.albicans.MethodsThe susceptibilitiesof 67C.albicansisolatestomethylrosaniliniumchloride, fluconazole, amphotericinB,itraconazole and ketoconazole were determined by using a broth dilution method as described in the National Committee for Clinical Laboratory Standards(NCCLS) document M27-A2.DNA was extracted from the C.albicans isolates and subjected to the amplification of 25S ribosomal DNA(rDNA) introns by PCR.The C.albicans isolates were divided into A,B and C genotypes according to the amplicons.The relationship was analyzed statistically by exact probability test between the genotypes and antifungal susceptibility of C.albicans.ResultsOf the 67 C.albicans isolates,8.96％ were resistant to fluconazole,2.98％ to itraconazole,1.49％ to ketoconazole,and no resistance to amphotericin B was observed.The minimum inhibitory concentration (MIC) of methylrosanilinium chloride varied from 0.125 to 4 mg/L.Statistical analysis revealed no significant correlation between the genotypes and autifungal susceptibilities of C.albicans(all P ＞ 0.05).Conclusions More attention should be given to the increasing resistance to fluconazole and itraconazole in some isolates of C.albicans as well as to the favorable antifungal activity of methylrosanilinium chloride.The antifungal resistance shows no apparent correlation to the genotypes of C.albicans.

Objective To investigate the early postoperative rehabilitation condition of patients with cervical spondylotic radiculopathy and analyze its correlation with the life style.Methods A retrospective analysis for 164 cases of cervical spondylotic radiculopathy with complete follow-up data were carried out.Using modified MacNab criteria for evaluation of postoperative recovery.The clinical efficacy was evaluated 3 months after operation,and the patients were divided into satisfactory group (excellent/good) and unsatisfactory group (fair/poor).The influencing factors included general information and the lifestyle such as smoking,drinking,sleep time,the height of the pillow,working time and frequency of physical exercise.The chi-square test and Logistic multivariate regression analysis were used to analyze the effects of lifestyle on early postoperative rehabilitation patients with cervical spondylotic radiculopathy.Results Postoperative efficacy satisfaction of patients evaluated by modified MacNab standard was 88.4％ in 3 months.There were differences in age(x2 =5.819),smoking(x2 =5.074) and frequence of physical exercise(x2 =4.430),and the differences were statistically significant (all P＜ 0.05).Logistic analysis showed that age more than 50 years old(OR=3.913,95％ CI =1.321-11.594,P=0.014),smoking(OR=3.217,95％ CI =1.126-9.195,P =0.029) and frequence of physical exercise less than or equal to 3 times a week (OR=3.906,95％ CI =1.026-14.869,P=0.046) were the risk factors affecting the satisfaction of clinical curative effect of early postoperative cervical spondylotic radiculopathy.Conclusion Age,smoking and physical exercise are related to early postoperative rehabilitation of patients with cervical spondylotic radiculopathy,and then more than 50 years old,smoking and exercise less than 3 times a week have adverse effects on the early postoperative rehabilitation.

ObjectiveTo investigate the role of apoptosis played in myocardial hypertrophy and the effect of L-arginine (L-Arg) on the pathogenesis.Methods36 rats were randomly divided into the control group, model group and L-Arg treating group. The animal model of over-loading myocardial hypertrophy was made, and systolic blood pressure (SBP)and the cardiac indexes were measured, spectrophotography and flow cytometry were used to detect the content of nitric oxide (NO), activity of superoxide dismutase (SOD) and apoptosis rate.ResultsIn the model group, SBP, cardiac indexes and myocardial apoptosis rate increased, the content of NO and activity of SOD decreased compared with the control group. While, in the L-Arg treating group, SBP, cardiac indexes and apoptosis rate decreased, the content of NO and the activity of SOD increased compared with the model group.ConclusionMyocardial apoptosis may play an important role in the pathogenesis of myocardial hypertrophy and cause the losing of myocardial cells. L-Arg induces the increasing production of NO and inhibits myocardial apoptosis through increasing the activity of SOD.

Objective To evaluate the effects of total glucosides of paeony (TCP) on cell proliferation of and expression of VECF and IL-23 in human HaCaT keratinocytes and their potential mechanisms. Methods MTT assay was performed to detect the cell proliferation of HaCaT cells incubated with various concentrations (0.5 to 312.5 mg/L) of TGP. HaCaT cells were classified into 8 groups, control group without any treatment, TGP groups treated with 6 different concentrations of TGP, SB203580 group treated with TGP of 125 mg/L after 2-hour pretreatment with SB203580 of 10 μmol/L After additional culture, reverse transcription (RT)- PCR and enzyme linked immunosorbent assay (ELISA) were conducted to determine the expression levels of VEGF and IL-23 mRNA and protein, Western blot to test the phosphorylation of p38 mitogen-activated protein kinase (MAPK) in these cells. Results The proliferation of HaCaT cells was promoted by TGP of low concentrations (0.5 and 2.5 mg/L), but inhibited by TGP of equal to or more than 12.5 mg/L, and peaked at the concentration of 125 mg/L. TGP of 0.5 and 2.5 mg/L enhanced the mRNA and protein expressions of VEGF and IL-23, while TGP of 12.5 to 125 mg/L suppressed the expression of VEGF mRNA and protein, and TGP of 62.5 to 125 mg/L downregulated the expression of IL-23 mRNA and protein. The phosphorylation of p38 protein kinase in HaCaT cells was induced by TGP of 125 mg/L in a time-dependant manner. Concretely, the level of phosphorylated p38 kinase in HaCaT cells was 0.3314 ± 0.0245 (peak) at 5 minutes, decreased to 0.2173 ± 0.0189 at 10 minutes (still statistically higher than untreated HaCaT cells) and 0.1664 ± 0.0201 at 30 minutes after treatment with TGP of 125 mg/L. SB203580 attenuated the effect of TGP on p38 phosphorylation, and the level of phosphorylated p38 kinase was 0.1529 ±0.0147 in HaCaT cells pretreated with SB203580 prior to the treatment with TGP. Conclusion TGP can inhibit the cell proliferation of and expressions of VEGF and IL-23 mRNA and protein in HaCaT cells, likely mediated by the p38 MAPK signaling pathway.

Objeetive To detect ERG11 gene mutations in clinical isolares of Candida albicans resistant to fluconazole.and discuss their relationship with formation of drug resistance.Methods Clinical specimens were collected.CHROMagar mediuln and amplification of the fragment spanning the conserved sequence of 25S rDNA including some transposable introns.were used to identify subtype Candida albicans isolates.FIuconazole sensitivity was detected in vitro through microdilution and Rosco tablets.The other three fragment of ERG11 gene were amplified and followed by sequencing with resistant type strain ATCC 76615-19 and Candida albicans Darlington strain with two sensitive isolates as controh.Results Fifteen resistant isolates of Candida albicans were found,all of which were type A.Sixteen silent mutations and 11 missense mutations were detected.Mutations in ATCC 76615-19 and Darlington strain were same with what had been reported.In the 2 sensitive strains.G640A(E165K),A945C(E266D)and G1609A/G(V488I)occurred,as well as the other 9 silent mutations.Only G487T(A114S)and T916C(Y257H)existed in each of 14 resistant isolates.In the other one resistant isolate,T541C(Y132H),T495A(D116E),A530C (K128T)and T1493A(F449Y)occurred Mong with 8 silent mutations.Conclusions The occurrence of G487T(A114S)and 1916C(Y257H)in 14 isolates from different sources suggested they may involve in fluconazole resistance.The novel mutation T1493A(F449Y)can appear in resistant isolves of Candida albicans.

BACKGROUND:Xenogenic deproteinized bone is generally collected and has unique biological properties;however,its immunogen hopes be resolved so as to discover a new type of bone graft material.OBJECTIVE:To explore the performance of xenogenic deproteinized bone scaffold for tissue-engineered bone and the effect on spinal intertransverse fusion in goats.DESIGN,TIME AND SETTING:An in vivo cell-scaffold study was performed at the Key Laboratory of Fibrotic Biotherapy of Heilongjiang Province between February and October 2008.MATERIALS:Twelve healthy male goats aged 6-8 months were provided by Animal Center of Mudanjiang Medical College.METHODS:Cancellous bone at distal femur of adult swine was obtained to prepare xenogenic deproteinized bone scaffold using physical and chemical methods.Effects of the scaffold on morphology,structure,component,biomechanical property,and biological behavior of seed cells were detected and analyzed.Bone marrow was extracted from ilium of goat and gradient-centrifuged to obtain the third-passaged bone marrow mesenchyme stern cells (BMSCs).A certain quantity of autologous BMSCs and recombinant human bone morphogenetic protein 2 (rhBMP-2) were plated onto the scaffold to achieve tissue-engineeried bone.Models of bilateral L3,4 intertransverse bone graft were established on 12 goats.Tissue-engineeried bone was implanted into the left side in a repairing group,while the same volume of autologous ilium was implanted into the right side in a control group.MAIN OUTCOME MEASURES:X-ray examination and histological detection were performed at 4,8,and 12 weeks after implantation.RESULTS:Deproteinization-treated cancellous bone exhibited a spatial grid structure of variously sized,crossing and opening pores,composing hydroxyapatite and collagen.The pore diameter was 200-500 μm,and porosity was about 60%.Mechanical property and cell compatibility were well.X-ray examination demonstrated that at 4 weeks after implantation,some or even lateral intertransverse bridging regions were unclear,and material density in the repairing group was lower than control group;at 8 weeks after implantation,interspace between up and down bridging was shrunk,and a great quantity of calluses were successively formed;at 12 weeks after implantation,complete confluence was observed,and the density in the repairing group was closed to that in the control group.Histological detection indicated that at 4 weeks after implantation,new bone was multidrop-formed in the repairing group;at 8 weeks after implantation,bone tissue like the islet shape was passed through the whole implanted materials;at 12 weeks after implantation,woven bones arranged across,medullary cavity was formed,and osteogenic activity in the repairing group was closed to that in the control group.On the other hand,at 4 weeks after implantation,a large quantity of new bones were formed in the control group;at 8 weeks after implantation,a great quantity of collagen fibers were observed,and osteogenesis was clear in marginal region;at 12 weeks after implantation,fibrous tissues were reduced,and osteogenesis was active.CONCLUSION:BMSCs and rhBMP-2 incubated on xenogenic deproteinized cancellous bone is an ideal tissue-engineeried bone scaffold,characterizing by an excellent histocompatibility and a strong osteogenesis,based on in vitro X-ray diffraction analysis,mechanical test,and in vivo osteogenesis experiment.

BACKGROUND:Compared with conservative treatment, percutaneous vertebroplasty (PVP) and percutaneous kyphoplasty (PKP) can rapidly relieve the pain caused by the compression of osteoporotic vertebral body. However, the perfusion of bone cement can not only restore the rigidity of fractured vertebrae, but also theoretical y alter the stress distribution of fractured vertebrae and increase the stress load of adjacent vertebrae, thus causing adjacent intervertebral disc degeneration. ＜br> OBJECTIVE:To observe the effects of PVP and PKP on the adjacent intervertebral disc degeneration and explore the possible mechanism. ＜br> METHODS:A total of 85 patients with osteoporotic vertebral compression fracture were divided into three groups:conservative treatment group (n=23), PVP group (n=42) and PKP group (n=23). ODI scores, DHI, MRII, and peripheral interleukin-1β, interleukin-6, tumor necrosis factor-αlevels were col ected and analyzed at different time points. ＜br> RESULTS AND CONCLUSION:The results showed that, PVP group and PKP group relieved pain effectively, reduced the time in bed, and had a better treatment satisfaction, but there were no significant differences in the scores of ODI scores between PVP group and PKP group at 24 months after discharge (P>0.05). At 24 months after discharge, MRII in the PVP group and PKP group were lower than that in conservative treatment group and before treatment (P＜0.05), while peripheral interleukin-1, interleukin-6, tumor necrosis factor-αlevels were higher (P＜0.05), there were significant differences between PVP group and PKP group (P＜0.05). The disc degeneration adjacent to the fractured vertebral body can be accelerated by PVP and PKP procedures, and the latter is more serious. Interleukin-1, interleukin-6, tumor necrosis factor-αlevels are closely related to the degree of intervertebral disc degeneration.

BACKGROUND:The inflammation of osteoarthritis is mediated by chondrocytes and the synovial tissue-secreted cytokines. Articular cartilage and synovial tissue contains a variety of cytokines, which play important regulatory role in the repair of articular cartilage injury.
OBJECTIVE:To analyze the relationship between chondrocytes, synovial tissue-secreted cytokines and osteoarthritis, as wel as the effect of chondrocytes and synovial tissue-secreted cytokines on osteoarthritis.
METHODS:A computer-based search was conducted in Wanfang database (www.wanfangdata.com.cn), PubMed database (www.ncbi.nlm.nih.guv/pubmed) by the first author from 2005 to 2010. The key words were“osteoarthritis, degeneration, cartilage tissue, cytokines”. A total of 146 articles were obtained through computer search, then reading the titles and abstracts for initial screen, 86 articles with the unrelated research purposes were eliminated, 40 articles with the repetitive content were excluded, and final y 21 articles related to the effect of various cytokines in the degenerative cartilage and synovial tissue of the osteoarthritis patients were included for further analysis.
RESULTS AND CONCLUSION:The cytokines mainly refers to the activated immune cel s and some stromal cel-secreted non-specific regulate immune responses and inflammatory responses-mediated smal molecule proteins, including lymphokines produced by the lymphocytes, as wel as the single nuclear factor and other cytokines produced by monocytes macrophages. Synovial cel-secreted cytokines may partial y explain the pathological process of osteoarthritis and play an important role in joint inflammation. Although, more and more scholars have pay attention to the effect of synovial cel s and cartilage cel-secreted cytokines, but mainly studies the effect of exogenous cytokines on chondrocytes or synovial cel s, and the effect of endogenous cytokines in the pathogenesis of osteoarthritis has not been extensively studied.

Objective To evaluate the possible roles of IL-18 in hepatitis B virus (HBV) infection. Methods Serum IL-18 level was measured using ELISA, and serum HBV DNA content was detected by FQ-PCR. Results There was significant difference in serum IL-18 level between the fulminant hepatitis patients and chronic hepatitis patients or normal controls (P