Objective: To study the protective effect of coenzyme Q_10 on red blood cell membrane and its immunity in patient during extracorporeal circulation. Method: Twenty patients under extracorporeal circulation about 30 minutes were randomly divided into control group and coenzyme Q_10 group (n=10). Coenzyme Q_10 2mg/kg was added to the priming fluid in coenzyme Q_10 group. Plasma levels of malondialdehyde (MDA), free hemoglobin (FHb), immune adhe sion ability and immune compound of red blood cell membrane were measured before extracorporeal circulation, 15 and 30 minutes after beginning of cardiopulmonary bypass, and on first day morning after operation. Result; All different periods after beginning of cardiopulmonary bypass in coenzyme Q_10, group, MDA and FHb levels were lower and im mune adhesion ability was higher than those of control group. Conclusion: Coenzyme Q_10 may protect red blood cell membrane and its immunity of patient during extracorporeal circulation.

Objective To evaluate the efficiency of PCR-restriction fragment length polymorphism (PCR-RFLP)aiming at the structure gene g6341,versus PCR fingerprinting analysis in the genotyping of Cryptococcus neoformans MethodsEight reference strains and 68 clinical and environmental isolates of C.neoformans were genotyped by PCR-RFLP and PCR fingerprinfing.In PCR fingerprinting,the minisatel lite-specific core sequence of wild-type phage M13 was used as a single primer.The structure gene g6341 was selected for PCR-RFLP analysis by sequence alignments of multiple genes,a pair of pnmers were developed based on the conserved region of g6341 gene.PCR products were digested with the appropriate restriction endonucleases,and RFLP profiles were analyzed.Partial sequence analysis of g6341 gene was performed for different genotypes of C.Neoformans.Phylogenetic analysis was done to study the relatedness between these genotypes.Results As sequence homology analysis showed,g6341 gene was suitable for RFLP analysis.In the case of enotyping of 76 C. Neoformans strains,the results obtained from PCR-RFLP were consistent with those from PCR fingerprinting.Sequence analysis of g6341 gene revealed a homology of 84%-97%among the eight genotypes as well as a consistency of 99%-100%within a same genotype.In the phylogenetic tree,genotypes VNⅠ,VNⅡ,VNⅢand VNⅣ belonged to one cluster,and genotypes VGⅠ,VGⅡ,VGⅢ and VGⅣ to another cluster.Conclusions PCR-RFLP analysis aiming at the structure gene g6341 is a useful tool to genotype C.neoformans.Sequence analysis of g6341 gene can disclose the relatedness among different molecular types of C.neoformans.

Objective：To study the ultrastructure changes of osteosarcoma cells(OS-732) inactivated by alcohol of two different concentrations and clinical application value.Methods：Osteosarcoma cells(OS-732) were inactivated by 75% and 95% alcohol and observed by light and electron microscope.And its viability was detected by MTT method. Results：Cell internal structure changed significantly and irreversible damage formed. MTT method proved that inactivated cells had no viability.Conclusion：These two inactivation methods were effective. Cell internal enviroment was damaged very seriously and cell was led to death. These two inactivation methods could provide choices for clinical limb protective operation.

Objective To investigate the molecular epidemiology of clinical cryptococcal isolates from China by analyzing the constituents and distributions of the varieties,genotypes and mating types (MAT)of them.Methods (1)PCR fingerprinting and PCR amplification were performed by using the minisatellite-specific core sequence of wild-type phage M13 as single primer.Genotypes of the 110 cryptococcal isolates from China were assigned by comparison with the reference strains of the 8 major molecular types loaded on gel.(2)Identification of the varieties and mating types was carried out by PCR using the specific primers of the varieties and mating types.Results Of the 110 clinical cryptococcal isolates,strains of Cryptococcus neoformans var.grubii with genetype VNⅠ and mating type MATα were the most representative ones(89.1%)followed by strains of C.neoformans var.gattii(8.2%)including isolates of genotype VG I,mating type MATα(7.3%)and genotype VGⅡ,mating type MATα(0.9%);AD hybrids with the genotype VNⅢ,mating type MAT-/α and genotype VN Ⅲ,mating type MATα/-(1.8%);and isolate of C.neoformans var.neoformans with the genotype VNⅣ and mating type MATa(0.9%).Conclusion Of the clinical isolates from China,all three varieties and AD hybrids are found.The vast majority(>99%) of strains possess the α allele in MAT locus and most of them are C.neoformans vat.grubii with the genotype VN I,which accord with the data of most studies of clinical molecular epidemiology in other geographic areas.However.no genotype of VNⅡ.VGⅢ and VGⅣ isolates are found in this study.

Objective To assess the subgenotypes of pathogenic Cryptococcus gattii isolates from China and to elucidate the epidemiological links between these domestic isolates and those from other parts of the world. Methods DNA was extracted from 9 clinical isolates of Ctyptococcus gattii from China. The partially variable regions of the three unlinked loci, namely IGS1, PLB1 and GEF1, were amplified and sequenced, and the bioinformation at these loci was obtained from GenBank for multi-locus sequences alignment and phylogenetic analysis. Results Of these 9 clinical isolates, 8 were genotype VG Ⅰ and mating type α with the same sequences at the tested regions as the reference strain WM276, which was a representative isolate of an independent subgenotype; 1 was of genotype VG Ⅱ and mating type α, which was the first report in China, with the tested sequences consistent with those of the referrence strain R272. Sequencing and phylogenetic analysis of GEF1 gene, which was located at mating type locus, successfully identified the genotypes and mating types of all the Cryptococcus gattii isolates involved here. Conclusions Multi-locus sequence analysis shows that causative Cryptococcus gattii isolates of genotype VG Ⅰ in China carry similar sequences at the tested loci in IGS1, PLB1 and GEF1 genes, to a widely distributed subgenotype in the world, and the sequences of the first VG Ⅱ genotype isolate from China resemble the less virulent subgenotype VG Ⅱ b found in Vancouver islands.

Objective To identify the AD hybrid strains and its hybrid types within Cryptococcus neoformans.Methods Difierent hybrid types of AD strains were analyzed by PCR 0f STE20 and MF genes within MAT locus and CIA4 and GPal genes out of MAT locus.The PCR-RFLP analysis of g6341 gene was also performed.Results The mating types of 18 AD strains were precisely identified by PCR of STE20 gene,whereas those of H strain were not identified.CL44 gene was better than the GPal gene in PCR identification of the AD hybrids.In the RFLP analysis of g6341 gene,AD strains were grouped into 2 distinct RFLP patterns based on the mating type on serotype A allele.The mating types of AD strains were not identified by the molecular analyses based on the CL44,GPal and g6341 genes.Conclusion It is necessary to use multi-locus analyses of genes within and out of the MAT locus in precise identification of the AD strains and their hybrid types of Cryptococcus neoformans.

Objective To evaluate the role of Restriction fragment length polymorphism (RFLP) analysis in detection of the fragment of GEF1α/a gene which are both located at ct and a mating type loci in identification of species, varieties, genotypes and mating types of Cryptococcus neoformans species complex(Cryptococcus neoformans and Cryptococcus gattii). Methods The GEF1α/a gene was selected from 20 genes which both located at α and a mating type loci for RFLP analysis, according to the requirements of sequence similarities and primer design in PCR-RFLP analysis. Primer pair was designed from the conserved regions of GEF1α/a genes of distinct genotypes and mating types of reference strains to amplify a fragment of GEF1α/a gene from Cryptococcus neoformans and Cryptococcns gattii strains tested. Sequence alignment,restriction maps analysis, endonucleases selection and electrophoresis stimulation were conducted by using DNAMAN and Vector NTI software. EeoT14 Ⅰ and Hap Ⅱ endonucleases were selected for RFLP analysis of the GEF1α/a fragments amplified from 125 isolates of Cryptococcns neoformans and Cryptococcus gattii. Results An approximate 1 300 bp fragment was amplified from total 82 Cryptococcus neoformans and 43 Cryptoceccus gattii isolates. However, negative PCR results were found in the reference strains of Cryptococcus laurentii, Candida albicans, Candida tropicalis, Candida parapsilosis, Candida krnsei,Candida glabrata, Trichosporon asahii, Aspergillus fumigatns and Aspergillus flavus. RFLP analysis successfully identified the species, varieties, genotypes and mating types of total 125 isolates of Cryptococcus neoformans and Cryptococcns gattii tested in this study. Condusion PCR-RFLP analysis of the GEF1α/a fragment has the potential value in identification of species, varieties, genotypes and mating types of Cryptococeus neoformans species complex simultaneously and rapidly, and may be a useful tool in molecular epidemiological analysis.

Objective To compare changes in indexes and analyze their values in prognosis of severe burn patients with sepsis.Methods A retrospective analysis was conducted. The patients with severe burn sepsis admitted to the Third Affiliated Hospital of Soochow University from August 2014 to December 2016 were enrolled. The blood culture was positive in the clinical diagnosis of sepsis. According to the prognosis, the patients were divided into death group and survival group. Their general information, vital signs, blood routine examination, serum sodium (Na+), serum glucose (Glu), C-reactive protein (CRP) and arterial partial pressure of carbon dioxide (PaCO2) at the time of admission and diagnosis of sepsis as well as the level of serum procalcitonin (PCT) at admission, diagnosis of sepsis and 1-8 days of post diagnosis were also compared. Receiver operating characteristic curve (ROC) was used to analyze the prognostic value of each index, and multivariate Cox regression analysis was used to analyze the influence of each index on the survival time, and the survival curve of Kaplan-Meier was analyzed for dead patients.Results There were 25 cases of severe burn patients with sepsis, which were admitted to hospital within 12 hours after injury; the time of diagnosis of burn sepsis was (14±6) days; 8 cases of survival; 17 cases died, the mortality rate was 68.0%, the time from diagnosis of sepsis to death was (28±14) days. The age of the death group was significantly higher than that of the survival group (years: 41±12 vs. 29±9,t = 2.598,P = 0.016), but there was no significant difference in the gender, total burn area,Ⅲ degree area, and the time of diagnosis of sepsis between the two groups. The platelet count (PLT) at the diagnosis of sepsis in death group was significantly lower than that of the survival group (×109/L: 69±43 vs. 180±108,t = -2.773, P = 0.023), and the PCT at 1-8 days of post-diagnosis in the death group was significantly higher than that of survival group [μg/L: 4.4 (2.2, 9.0) vs. 1.6 (0.7, 2.3),Z = -2.521,P = 0.012], but there was no significant difference in body temperature, heart rate, white blood cell count (WBC), percentage of neutrophils (Neu), Na+, Glu, CRP, PCT, PaCO2 at the time of admission and diagnosis of sepsis and PLT at the time of admission between the two groups. ROC curve analysis showed that the area under ROC curve (AUC) of age, PLT at the time of diagnosis and PCT at 1-8 days of post-diagnosis of sepsis was 0.808, 0.779, 0.825, respectively, for predicting the prognosis of patients with severe burn sepsis (allP < 0.05). At the cut-off age of 32, the sensitivity was 73.3% and the specificity was 75.0%. As the cut-off of PLT was 138×109/L at the time of diagnosis, the sensitivity was 92.3% and the specificity was 75.0%. As the cut-off of PCT was 2.39μg/L at 1-8 days of post-diagnosis of sepsis, the sensitivity was 73.3% and the specificity was 87.5%. Multivariate Cox regression analysis showed that age and PLT at the time of diagnosis were the favorable factors for the survival time of patients with severe burn sepsis (β value were -1.834, -0.029, respectively, bothP < 0.05). Kaplan-Meier survival analysis for patients in the death group showed that the median survival time of patients ≥32 years old was longer than that of patients < 32 years old (days: 32 vs. 9); 18-day cumulative survival rate was significantly higher than that of patients < 32 years old [83.3% (10/12) vs. 25.0% (1/4),χ2 = 9.705,P = 0.003].Conclusion Age, PLT at diagnosis of sepsis and PCT at 1-8 days after diagnosis of sepsis could be used as prognostic indexes for severe burn patients with sepsis.

OBJECTIVETo explore the causes, prevention, and management of the complications during intracranial aneurysm embolization with controllable coils (mechanical detachable spiral, MDS; and Guglielmi detachable coil, GDC).

METHODSRetrospective review of 120 cases with 125 intracranial aneurysms embolized with controllable coils from March 1995 to July 1999 was conducted. The 20 accidents (in 18 cases) including aneurysm rupture, over-embolization, protrusion of coil end into the parent artery, and thrombosis of the parent artery were analyzed.

RESULTSAmong the 20 accidents, there were 6 aneurysm ruptures, 6 over-embolizations (in 5 cases), 6 coil protrusions, and 2 thromboses (one was secondary to coil protrusion). The embolization-related mortality was 3.33% (4/120), the permanent neurological deficit was 1.67% (2/120), and the transitory neurological deficit was 3.33% (4/120). The occurrence and outcome of the complications were related to the embolizing technique, the pattern of aneurysm and its parent artery, the imperfection of embolic materials, and the observation and management during embolization.

CONCLUSIONSkilled embolizing technique, better understanding of the angio-anatomy of an aneurysm and its parent artery, correct judgement and management during embolization, and improvement of embolic materials are beneficial to the reduction of complications and to the melioration of the outcome of complications.

Aneurysm, Ruptured ; etiology ; Embolization, Therapeutic ; adverse effects ; instrumentation ; Humans ; Intracranial Aneurysm ; therapy ; Retrospective Studies ; Thrombosis ; etiology

Objective To determine in vitro drug susceptibility to five antifungal agents of clinical Cryptococcus neoformans strains isolated from different areas of China in recent ten years. Methods Eighty clinical isolates of Cryptococcus neoformans were isolated from Shanghai, Guangdong, Fujian, Beijing and some other areas of China from 1998 to 2007. The minimal inhibitory concentrations (MIC) of the isolates to five antifungal agents, including amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole, were determined using broth microdilution procedure (document M27-A2) recommended by the Clinical and Laboratory Standards Institute (CLSI). Kruskal-Wallis rank sum test was employed for the statistical analysis. Results The MIC50 of the Cryptococcus neoforrnans isolates tested for amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole were 0.5, 4, 2, 0.25 and ≤0.031 3 mg/L, respectively; and the MIC<,90> of the isolates tested for the above antifungal agents were 1, 8, 4, 0.5 and 0.062 5 mg/L, respectively. Among the tested isolates, 3 (3.8 %) were resistant to flucytosine, 4 (5.0 %) were resistant to itraconazole. All isolates were susceptible to amphotericin B and voriconazole. There was no significant difference in MIC of the strains isolated from any particular years to the five agents (χ2=0.500,2.687,2.211, 2.660,0.677,P>0.05). Conclusions The Cryptococcusneoformans isolates are highly susceptible to the five antifungal agents, while a few strains are resistant to flucytosine or itraconazole. The drug susceptibilities of the strains isolated from particular years are similar.