To investigate the role of ERC,6 gene in the growth rate and antifungal susceptibility,Aspergillus fumigatus strain with extra copies of ERG6 gene was constructed.Open reading frame (ORF) of putative ERG6 gene was searched in A.fumigatus genome.A PCR fragment, ERG6 ORF sandwiched by its flanking sequences (about 1 kb respectively),was amplified and was then subcloned into vector pRG-AMA1- NotI to produce a recombinant plasmid pERG6,which was further transformed into uracil auxotroph A.fumigatus strain AF293.1 to produce the transformant AF-pERG6.In the same time,empty plasmid pRG-AMA1-NotI was also transformed into A.fumigatus strain AF293.1 to produce the transformant AF-empty as a control.Radial growth of the transformants was tested on minimal medium (MM) and YAG medi- um.Antifungal susceptibilities of these resahing transformants,AF-pERG6 and AF-empty,to the common antifungal agents were performed by using both disk diffusion and broth microdillution methods.A.fumigatus genome contains a ERG6 gene,of which the ORF size is 1 256 bp.Comparing to Candida albicans and Sacchromyces cerivisiae sterol methyltransferase,A.fumigatus Erg6p had 57% and 50% identity, and had 70% and 63% similarity in amino acid sequences,respectively.Radial growth of transformant AF-pERG6 was slower than that of transformant AF-empty.The antifungal susceptibilties of transformant AF-pERG6 to the antifungal drugs itraconazole,voriconazole,terbin- aline,amphotericin B,caspofungin and grisefolvin were same to that of transformant AF-empty.In A.fumigatus,extra copies of ERG6 gene have no effect on antifungal susceptibilities to itraconazole,voriconazole,terbinafine,amphotericin B,caspofungin and grisefolvin.Radial growth of A.fumigatus harboring extra copies of ERG6 gene becomes slower compared to the control.

In recent years,with the increasing of immunocompromised patients in clinical practice,invasive candidiasis has been increased accordingly. Although several antifungal drugs,including polyenes,azoles,5-flucytosine,and echinocandins,have been used widely to treat invasive candidiasis,the treatment failure that caused by the resistance of Candida albicans to these antifungal drugs have been reported widely. Here,we reviewed the molecular mechanisms of resistance to azoles,polyenes,5-flucytosine,and echinocandins,such that to provide an update on these issues.

Objective To evaluate the effects of endogenous NO on the chemosensitivity of human breast cancer cell. Methods MCF-7 cells were cultured as monolayer, incubated with cytokine IL-1β. The pro-duction of NO was detected by NO assay. The expression of iNOS protein was measured by Western blotting. Establishing control group and experimental group, the chemosensitivity of MCF-7 cells incubated by L-NMMA and L-Arg to ADM and 5-Fu was studied by MTT assay. Results There was a positive correlation of dose-dependence between NO production and IL-1β concentration. MCF-7 cells expressed plenty of iNOS by induetion of IL-1β. There was no significant difference on iNOS whether L-NMMA and L-Arg existed or not. Incubating MCF-7 cells with 0. 5 μmol/L or 1 μmol/L ADM, the survival rate of experiment group was remarkablely decreased(P < 0.05) ; L-NMMA significantly increased survival rate of experiment group(P < O. 05) ; L-Arg decreased survival rate of experiment group(P < 0.05). Conclusion The induction of IL-113 in MCF-7 cells can increase the production of endogenous NO, which increases MCF-7 cells' sensitivity to chemotherapy.

OBJECTIVETo explore the treatment of chronic calcaneal osteomyelitis with bone defect after debridement and evaluate its clinical outcomes.

METHODSFrom June 2009 to June 2011, 52 patients with chronic calcaneal osteomyelitis were treated with stage-one compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone,including 12 females and 40 males with an average age of 43 years old ranging from 18 to 67. According to Cierny-Mader classification, there were 34 cases with stage III and 18 with stage IV. There were 32 cases on right side and 20 on left,with a course of 6 months to 3 years. The area of soft tissue wound ranged from 3.0 cm x l.5 cm to 23.0 cm x l2.0 cm. The clinical effects were evaluated according to infection controlling, calcium sulfate absorption,bone defect repair and heel functional recovery.

RESULTSAll patients were followed up for 2 to 3.8 years (averaged 2.8 years). Primary healing was achieved in 52 patients. Two cases of recurrence were found post-operatively, 1 case in 3 months and another in 5 months,which were cured after a second operation. Bone repair healing was gained in 1.5 to 3.5 months (averaged 2.5 months). Complete radiological absorption of calcium sulfate was found in 1.2 to 3 months(averaged 2.2 months). Local exudation after removal of drainage tube had been persisting in 10 patients for 2 to 3 months, which was consistent with the time when cacium sulfate were totally absorbed. Flap had partial necrosis in 4 cases,and the wounds were closed after appropriate treatment finally. The mean Maryland score was 88.15±7.70. There were excellent results in 32 cases, good in 14, fair in 6.

CONCLUSIONA satisfactory short-term clinical results can be gained by one-stage compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone in chronic calcaneal osteomyelitis, but the long-term results need further follow-up. And much more study is also demanded to reduce the exudation of calcium sulfate.

Adolescent ; Adult ; Aged ; Anti-Bacterial Agents ; therapeutic use ; Bone Transplantation ; Calcaneus ; injuries ; surgery ; Calcium Sulfate ; therapeutic use ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Osteomyelitis ; drug therapy ; physiopathology ; surgery ; Surgical Flaps ; Wound Healing ; Young Adult

Objective To evaluate the susceptibility of itraconazole-resistant Aspergillus fumigatus isolates to four other antifungal drugs.Methods Six strains of Aspergillus fumigatus were serially isolated from a patient with lung aspergilloma,who was resistant to itraconazole therapy.Clinical Laboratory Stan- dard Institute broth microdilution method M38-A and E-test were applied to test the susceptibility of A.fumi- gatus strains to amphotericin B,itraconazole,voriconazole,caspofungin and micafungin.Results With M38-A method,the minimal inhibitory concentration(MIC)of itraconazole was 0.5?g/mL for 2 strains and more than 16?g/mL for the other 4 strains;the MICs of amphotericin B and voriconazole were 1?g/mL and 0.25-1?g/mL respectively for all the 6 strains;and the minimal effective concentration(MEC)of caspofungin and micafungin were no more than 0.03?g/mL for all the 6 strains.Moreover,E-test results also revealed that caspofungin and voriconazole had a good antifungal activity against all the strains,including the 4 itraconazole-resistant ones.Conclusion The itraconazole-resistant A.fumigatus strains isolated from a patient with lung aspergilloma are susceptible to amphotericin B,voriconazole,caspofungin,and micafungin.

To systematically evaluate the clinical efficacy and safety of compound Danshen injection in treating hypoxic-ischemic encephalopathy (HIE) of newborns. Computer retrievals were made in PubMed, Embase, Cochrane Library, CBM, CNKI, VIP and China info (before May 2014) and relevant literature references, and manual retrievals were made for journals and conference papers, in order to collect randomized or semi-randomized controlled trials concerning compound Danshen injection in the treatment of neonatal HIE. The quality of included references was evaluated according to literatures recommended by Cochrane Handbook. RevMan 5. 3 software was applied in the statistical treatment. Finally, a total of 13 randomized controlled trials were included, covering 1,211 patients (including 639 patients in the compound Danshen injection-treated group and 572 patients in the control group). Meta-analysis results showed that the routine treatment combined with compound Danshen injection can improve the treatment efficiency of neonatal HIE [RR = 1.28; 95% CI (1.21-1.36)], reduce the mortality rate [RR = 0.42; 95% CI (0.23-0.75)] and the incidence of long-term neurological sequelae [RR = 0.48; 95% CI (0.35-0.65)], with statistical differences. No fatal side effect was observed in all of included trials. So far, limited evidences in this study proved that the application of compound Danshen injection in the treatment of neonatal HIE can enhance the clinical efficiency. However, because of the low quality of the included trials, more well-designed and large-scale multi-center randomized controlled trials shall be made in the future.
Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypoxia-Ischemia, Brain ; drug therapy ; Infant, Newborn ; Infant, Newborn, Diseases ; drug therapy ; Injections ; Randomized Controlled Trials as Topic

Objective To study the nasal mucosa absorption of paeonol ofXingbi microemulsion-based thermo-sensitive gel in rats; To evaluate the nasal mucosa absorption of medicine and the rationality of preparation form.Methods According to the nasal administration method, the blood medicine concentration of paeonol was determined at different time points.Results The nasal administration bioavailability of paeonol in Xingbi microemulsion-based thermo-sensitive gel was 78.68%, the plasma concentration reached the maximum (0.3404 μg/mL) after 3 min of administration, and the average residence time was 9.23 h. After reaching Cmax, The plasma concentration changed similar to intravenous administration of paeonol. The concentration data of paeonol in plasma was consistent with a two-compartment model, with a weight of 1.Conclusion Xingbi microemulsion-based thermo- sensitive gel has a higher bioavailability, and the preparation form was reasonable.

Objective The objective of this study was to investigate the miR-345 expression level in non-small cell lung cancer(NSCLC)tissue and its potential clinical significance.Methods Real-time PCR was conducted to evaluate the expression of miR-345 in NSCLC tissues in 98 samples and cell lines.Then the association between tissue miR-345 expression level and clinical outcomes was further analyzed.Results The expression level of miR-345 was significantly decreased in NSCLC tissues and cell lines when compared to the controls(P<0.05).miR-345 expression level in NSCLS tissues was associated with various clinicopathological parameters including LN metastasis(P<0.001),distant metastasis(P=0.028),TNM stage(P=0.004)and grade(P=0.011).In addition,the NSCLC patients in the low miR-345 expression group showed significantly shorter overall survival time than that in the high miR-345expression group(P=0.021).Multivariate analysis showed that the expression of miR-345 was an independent risk factor for NSCLC(HR=3.897,95% CI:2.263~10.440;P=0.012).Conclusion The expression level of miR-345 was decreased in NSCLC tissues as well as cell lines compared with normal controls.Low miR-345 expression in tissues was associated with progression and poor prognosis of NSCLC.These results indicate that miR-345 may be a novel prognostic marker in NSCLC.

Objective To explore the test-retest reliability of the open field test,as a method for ethology in Kunming mice. Methods The behavior of adult male Kunming mice in the open field test was recorded twice,once for five minutes, with a one-week inter-trial interval. The following parameters were evaluated:number of central squares; number of peripheral squares; percent of central squares; number of horizontal squares; number of rearings; total score; and number of fecal boli. Subsequently,the intraclass correlation coefficient (ICC) and the agreement index (Kappa) were calculated for each of these parameters. Results Both ICC and Kappa for total score( ICC =0.902, P＜0.01; Kappa =0.680, P＜0.01; r=0.815, P＜0.01 ) showed a good reliability and correlation between the test trials. Only ICC for number of central squares ( ICC = 0. 578, P ＜ 0.01 ), number of peripheral squares( ICC =0.615, P＜0.01 ) ,percent of central squares(ICC=0.523, P＜0.01) ,number of horizontal squares( ICC = 0. 607, P ＜ 0.01 ) and number of rearings ( ICC = 0. 606, P ＜ 0.01 ) indicated a fair reliability between the test trials. Neither ICC nor Kappa for number of fecal boli displayed a significant reliability between the test trials. The difference of these parameters between test and retest, such as number of peripheral squares( t =3.01, P＜0.01 ),number of horizontal squares( t = 3.34, P ＜ 0.01 ) and total score( t = 7.86, P ＜ 0.01 ) were significant. Conclusion The parameter,total score for Kunming mice in the open field test,is found to be stable over time, and indicates a good reliability with a one-week inter-trial interval.

Objective To explore the behavioral dimensions and validity of the light-dark box(LDB),as an model of state anxiety in Kunming mice.Methods The behavior of adult,male,Kunming mice in LDB was recorded twice,for five minutes,with a one-week inter-trial interval.The following parameters were evaluated by factor analysis:latence for the first entry into the dark area(Latence);percentage of time exploring in the light area (Ltime%);percentage of squares crossing in the light area(Lcross%);percentage of rears in the light area (Lrear%);transitions between two areas(Transition);total number of squares crossing in the whole apparatus (Cross);total number of rears in the whole apparatus(Rear);and number of fecal boli(FB).Results There was a good intercorrelation both among Ltime%,Lcross%,Lrear%,and among Transition,Cross,Rear.A good correlation also was found between test and retest for Transition,Rear and FB.Ltime%,Lcress%and Lrear%had significant loadings on Anxiety factor(Loading%:39.54%,35.40%);Transition loaded on Locomotor factor(Loading%:23.78%,19.66%),combined with Cross;Rear loaded on Exploration factor(Loading%:13.75%,22.60%);and FB loaded on Emotionality factor(Loading%:14.42%,13.02%);Latence was not stable.Conclusion Light-dark box,as an animal model of state anxiety in Kunming mice,comprised three behavioral dimensions:anxiety factor,LDB-activity factor(locomotor factor and exploration factor)and emotionality factor.