Objective To investigate the molecular mechanisms of cross-resistance to azoles in a clinical isolate of Aspergillus fumigatus. Methods A. fumigatus was isolated from a patient with invasive aspergillosis.Clinical Laboratory Standard Institute M38-A2 broth microdilution method and E-test method were used to determine the minimum inhibitory concentrations (MICs) or minimum effective concentration (MEC) of itraconazole, voriconazole, amphotericin B, posaconazole and caspofungin for the A. fumigatus isolate. DNA was extracted from the isolate and subjected to the amplification of cyp51A gene encoding the target enzyme of azole antifungal agents followed by sequence analysis. Results The broth microdilution test showed that the MEC of caspofungin was 0.5 mg/L, and MICs of itraconazole, voriconazole and amphotericin B were ≥ 16 mg/L,8 mg/L and 1 mg/L, respectively, for this isolate; while E-test assay revealed that the MICs of caspofungin,itraconazole, voriconazole, amphotericin B and posaconazole were 0.047 mg/L, ≥32 mg/L,≥32 mg/L, 12 mg/L and ≥32 mg/L, respectively. Sequence analysis showed an insertion of a 34-bp tandem sequence in the promoter region of the cyp51A gene as well as a T364A point mutation causing the substitution of leucine 98 (L98H). In addition, there were some other mutations in the cyp51A gene of this isolate, such as A137T,G585A, C814A, G836C, T991C and A1350G, which could result in corresponding amino acid substitutions.Conclusions An A. fumigatus strain with cross-resistance to azole antifungal agents is isolated. There is an insertion of a 34-bp tandem sequence into the promoter region as well as a T364A point mutation in the cyp51A gene, which contribute to the cross resistance to azole antifungal agents including itraconazole, voriconazole,and posaconazole. In addition, other mutations causing amino acid substitutions have also been detected in the cyp51 A gene of this isolate.

Two patients were admitted to the hospital for 2-month history of pruritic eruptions on the forehead and 2-week history of pruritie eruptions on the leg, respectively. Both patients had a history of pet contact. Topical application of glucocorticoids did not work well. Dermatological examination revealed a patch measuring 5 cm ×6 cm on the forehead of one patient and a patch measuring 2 cm × 3 cm on the leg of the other patient. Both patches were surrounded by red papules and scaling. Microscopic examination of skin scales revealed hyphae and chain-like spores, and culture of skin scales grew Microsporum gyeum. Both the isolates of Microsporum gyeum were sensitive to ketoconazole, miconazole, bifonazole, terbinafine, and voriconazole. Both patients were healed after treatment with oral terbinafine and topical ciclopirox olamine.

Objective To investigate the in vitro susceptibility of 16 strains of Exophiala dermatitidis to 6 commonly used antifungal agents. Methods The Glinical and Laboratory Standards Institute (CLSI)M27-A2 protocol was carried out to determine the MIGs of terbinafine (TRB), itraconazole (ITC), amphotericin B (AMB), fluconazole (FLC), voriconazole (VRC), and caspofungin (GAS) to 16 strains of E. dermatitidis, and E-test was performed to determine those of VRG, ITC and AMB. Besides, the minimal fungicidal concentrations (MFGs) of the above antifungal agents to the 16 strains of E. dermatitidis were further assessed.The activity of TRB in combination with ITC and VRG against E. dermatitidis was also estimated. Results The MIC ranges of TRB, VRC, ITC, AMB, FLC, and CAS were 0.125 - 0.25 mg/L, 0.25 - 0.5 mg/L, 2.0 mg/L,2.0 mg/L, 16 - 32 mg/L and 16 - 64 mg/L respectively as shown by M27-A2 microdilution assay, while the MIC ranges of VRG, ITG and AMB, as determined by E-test, were 0.032 - 0.094 mg/L, 0.047 - 0.5 mg/L and 0.125 - 3.0 mg/L, respectively. The MFC ranges of TRB, VRC, ITG, AMB and FLG were 0.125 - 0.5 mg/L,0.25 - 0.5 mg/L, 2.0 - 4.0 mg/L, 2.0 - 4.0 mg/L and 16 - 64 mg/L, respectively. No synergism in the acitivity against E. dermatitidis was observed for the combination of TBR with ITC or VRC. Conclusion E. dermatitidis is susceptible to TRB, ITC, AMB, and VRC, but less sensitive to both FLC and GAS.

AIM: To investigate the postoperative changes of binocular visual function in patients with congenital superior oblique palsy.METHODS: Eye position and binocular visual function were examined in 28 patients with congenital superior oblique palsy pre- and post-operatively. The results were statistically analyzed.RESULTS: The normal eye position rate after operation was 89.3%. There were 9 patients who gained simultaneous perception after operation, while 13 patients gained fusion as many as that gained stereoacuity. There was statistically significant difference between postoperative and preoperative eyes (P<0.05). Near stereoacuity was improved after operation. Patients who had better visual acuity and low strabismus degrees as well as fusion had a better recovery.CONCLUSION: The operation could improve the eye position and rebuild the binocular visual function of the congenital superior oblique palsy. The recovery of binocular visual function was related to visual acuity and strabismus degrees of the patients and whether they had fusion function.

Objective To evaluate the performance of VITEK 2-Compact GN13 methods for testing imipenem susceptibility of Klebsiella pneumoniae and assess the reliability of its Advanced Expert System (AES).Methods A retrospective study was conducted with a total of 157K. pneumoniae strains, which were isolated from blood and intra-abdominal infections in the First Affiliated Hospital of Nanchang University from 2014 to 2015. Thein vitro minimum inhibitory concentration (MIC) values of imipenem were determined by disc diffusion, VITEK 2-Compact GN13 and broth microdilution methods, respectively. Categorical agreement (CA) rates of disc diffusion and VITEK 2-Compact GN13 methods were determined using broth microdilution as reference method. The genes encoding ESBLs and carbapenemase were screened by PCR and sequencing analysis. The phenotypic confirmatory tests such as modified Hodge test, PCR and DNA sequencing were used to confirm the resistance mechanism and evaluate the reliability of AES in interpreting the imipenem susceptibility of K. pneumoniae.Results Among the 157 isolates, 64 and 8 were identified as resistant and intermediate strains by broth microdilution method, respectively; 52 and 10 were tested as resistant and intermediate strains by disc diffusion method, respectively; 54 and 13 were determined as resistant and intermediate strains by VITEK 2-Compact GN13 method, respectively, while 70 and 3 were judged as resistant and intermediate strains by VITEK 2-Compact GN13 method plus AES validation. The CA of disc diffusion and VITEK 2-Compact GN13 methods compared with broth microdilution method were all higher than 90 %. However, the major error (ME) rate was 3.8 % and very major error (VME) rates were all 0.6 % in imipenem susceptibility testing by VITEK 2-Compact GN13 and disc diffusion. The imipenem susceptibility of 16 strains were modified by the AES, which eliminated 0.6 % VME, but increased major error by 1.3 % and minor error by 1.9 %. Phenotypic confirmatory tests showed that 75 % (12/16) of these strains were validated as producers of both ESBLs and carbapenemase, which was consistent with the result of AES validation. PCR and DNA sequencing analysis proved that 62.5 % (10/16) of these strains produce IMP-4/KPC-2 /NDM-1 and ESBLs.Conclusions Both disc diffusion and VITEK 2-Compact GN13 methods can be used for testing the imipenem susceptibility of K. pneumoniae isolates with reliable and accurate results. Attention should be paid to the possibility of ME and VME when testing imipenem susceptibility. The VME can be avoided by the AES mechanism. However, AES intervention will increase ME and minor error, which may be associated with decreased expression of carbapenemase.

Objective To evaluate the effect of using serf-designed facial support in prone position for patients after vitrectomy and silicone oil tamponade. Methods 100 patients with vitrectomy and sili-cone oil tamponade were randomly divided into the experimental group and the control group with 50 pa-tients in each group. The self-designed facial support in prone position was used in the experimental group, while the routing nursing measure was used in the control group. The ocular adverse effect and re-lapse rate were compared between the two groups. Results The incidence rate of corneal edema, in-creased intraocular pressure and retinal redetachment in the experimental group was higher than those in the control group, the time of keeping position in the experimental group was longer than that in the con-trol group. Conclusions Self-designed facial support in prone position can decrease discomfort and pain caused by compulsive position, reduce the incidence of complications, improve retinal attachment and increase the successful rate for the patients with vitrectomy and silicone oil tamponade.

to common antifungal drugs.

Objective:To investigate the efficiency of Agrobacterium tumefaciensmediated transfor-mation of Aspergillus fumigatus by using pyrG as a recessive selectable marker.Methods: FAP1 and SHO1 genes target sequences,composed of a selectable marker pyrG and the flanking sequences of the FAP1 and the SHO1 genes,were cloned into a binary plasmid pDHt/sk,respectively.The produced plasmids were transformed into A.tumefaciens.The A.tumefaciens and uracil auxotroph A.fumigatus were cocultured in induction medium without uricil and uridine at 24 ℃ for 48 h.To inhibit growth of A.tumefaciens and to select transformants,the cultures were transferred to 37 ℃ and incubated for another 48 h.Results: In this study,A.tumefaciens-medidated transformation of A.fumigatus produced high homologous recombination rates,which was 44%(7 of 16) for FAP1 and 35%(7 of 20) for SHO1.Conclusion: Our study showed that A.tumefaciens-medidated transformation by using pyrG as a recessive selectable marker is an efficient tool for target gene deletion of A.fumigatus.

Objective To study the effect of clinical nursing path with graph and text instructions (GTI) on health education to patients with complex retinal detachment.Methods Toally 76 inpatients with complex retinal detachment were enrolled in the study:38 of them hosptalized from March to August 2015 were put in the control group received routine treatment and the others hosptalized from September 2015 to March 2016 as the experiment group treated with GTI.The clinical outcomes were compared.Result The patients treated with GTI were better in the knowledge of the complex retinal detachment and moreover,they felt significantly more satisfied compared to that of the control group (P＜0.05).Conclusion It is beneficial to use GTI in the patients with complex retinal detachment.GTI is effective for improving their knowledge on the disease,and increasing their satisfaction with the clinical nursing.