Effects of microwave exposure on the enzymatic activity of Trichophyton rubrum / 中华物理医学与康复杂志

Objective@#To investigate the effects of exposing Trichophyton rubrum fungus to microwaves at different intensities in terms of the activity of succinic dehydrogenase and beta-(1, 3)-D-glucan synthase.@*Methods@#Trichophyton rubrum organisms were randomly divided into a control group and experimental groups. The experimental groups were incubated at 27 ℃ after direct radiation with 2450 MHz microwaves at 20, 40, 60 or 80 W for 15 min, repeated 8 times. The control group was incubated without any irradiation. Thirty days later, the beta-(1, 3)-D-glucan synthase and succinate dehydrogenase activities were determined using enzyme-linked immunosorbent assays.@*Results@#The enzymatic activity decreased gradually with increasing radiation intensity. When the output power was 80 W, the beta-(1, 3)-glucan-synthase-D activity was 0.730±0.74 U/ml and that of the succinate dehydrogenase was 1.828±1.774 U/L, both significantly lower than in the groups subjected to less powerful irradiation.@*Conclusions@#Microwave radiation can decrease the enzymatic activity of Trichophyton rubrum in a dose-dependent manner. Higher intensity is more effective. Microwave irradiation can decrease the activity of succinate dehydrogenase and beta-(1, 3)-glucan synthase from Trichophyton rubrum in vitro, resulting in the destruction of fungal cell walls and interfering with the tricarboxylic acid cycle, furthering cell death. Moreover, the temperature change possibly also helps promote the biological effects of microwave radiation.

Effects of microwave exposure on the enzymatic activity of Trichophyton rubrum / 中华物理医学与康复杂志

Objective To investigate the effects of exposing Trichophyton rubrum fungus to microwaves at different intensities in terms of the activity of succinic dehydrogenase and beta-( 1, 3)-D-glucan synthase. Methods Trichophyton rubrum organisms were randomly divided into a control group and experimental groups. The experimental groups were incubated at 27 ℃ after direct radiation with 2450 MHz microwaves at 20, 40, 60 or 80 W for 15 min, repeated 8 times. The control group was incubated without any irradiation. Thirty days later, the beta-( 1, 3)-D-glu-can synthase and succinate dehydrogenase activities were determined using enzyme-linked immunosorbent assays. Results The enzymatic activity decreased gradually with increasing radiation intensity. When the output power was 80 W, the beta-(1,3)-glucan-synthase-D activity was 0.730±0.74 U/ml and that of the succinate dehydrogenase was 1.828±1.774 U/L, both significantly lower than in the groups subjected to less powerful irradiation. Conclusions Microwave radiation can decrease the enzymatic activity of Trichophyton rubrum in a dose-dependent manner. Higher intensity is more effective. Microwave irradiation can decrease the activity of succinate dehydrogenase and beta-( 1, 3)-glucan synthase from Trichophyton rubrum in vitro, resulting in the destruction of fungal cell walls and interfering with the tricarboxylic acid cycle, furthering cell death. Moreover, the temperature change possibly also helps promote the biological effects of microwave radiation.

The value of combined detection of LTB4 and LTFA in early diagnosis of renal involvement in children with henoch schonlein purpura and its influence on prognosis / 国际检验医学杂志

Objective To investigate the value of leukotriene B4(LTB4)and leukotriene E4(LTFA)in early diagnosis of renal involvement in children with Henoch Schonlein purpura(HSPN)and its influence on prognosis.Methods A total of 185 children with HSPN were enrolled in our hospital from January 2014 to October 2016.A total of 50 healthy children were selected as control group at the same period.The serum levels of LTB4 and LTFA in all subjects were detected,and their value in early diagnosis of HSPN and its influence on prognosis were analyzed.Results Compared with the control group,the serum levels of LTB4 and LT-FA in children with HSPN were significantly higher(P<0.05),the contents were 987.6 ng/L and 896.3 ng/L,respectively.The AUC of the combined detection of HSPN was 0.899 and the sensitivity was 0.810,which was better than that of LTB4 and LTB4 alone(P<0.05).The average follow-up was(28.4 ± 10.3)months,with grade A prognosis in 133 cases(71.9%),B grade in 47 ca-ses(25.4%),and C grade in 5 cases(2.7%).The rank sum test showed,the prognosis of children with LTB4<987.6 ng/L was better than that of children with LTB4≥987.6 ng/L,the prognosis of children with LTFA<896.3 ng/L was better than that of children with LTFA≥896.3 ng/L.Spearman analysis showed that the content of LTB4 and LTFA was negatively correlated with prognosis(Rs= -0.693 and -0.637,P<0.05).The higher contents of LTB4 and LTFA,the worse the prognosis.Conclusion Combined detection of LTB4 and LTFA has important clinical significance for early diagnosis and prognosis of HSPN.

Establishment of digital polymerase chain reaction-ribotyping and database for Clostridium difficile genotyping / 中华传染病杂志

Objective To develop a digital polymerase chain reaction (PCR) ribotyping method and database for Clostridium difficile genotyping.Methods Sequencer based fluorescence capillary gel electrophoresis was used,instead of agarose gel electrophoresis,to establish the digital PCR-ribotyping of Clostridium difficile.Forty Clostridium difficile reference strains,consisting of 10 PCR-ribotypes (RT),were genotyped by the new digital PCR-ribotyping method to set-up the database.Results The sequencer based fluorescence capillary gel electrophoresis correctly detected PCR-ribotyping products of the 40 reference strains,and showed as digital figure;significant differences of these digital figures were found between the 10 RT.High similar digital figures were shown in twenty-one RT027 strains,three RT002 strains and two RT014 strains.However,seven RT001 strains were typed as four subtypes,and two RT014 strains as two subtypes,respectively.Conclusion A digital PCR-ribotyping,and a reference database consisting of 10 RT are successfully established.

Detection of genotype and toxin associated gene of hypervirulent Clostridium difficile clinical isolates / 国际检验医学杂志

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .

Detection and genotyping of moxifloxacin-resistant Clostridium difficile by multiplex polymerase chain reaction / 中华传染病杂志

Objective To develop a multiplex polymerase chain reaction (PCR )method for detecting and genotyping moxifloxacin-resistant Clostridium difficile (C.difficile)isolates.Methods Specific PCR primers of slpA genotypes gr,hr,fr,gc08 and 078 were designed according to the differences of slpA nucleotide sequences in different C.difficile genotypes,and the house-keeping gene tpi specific PCR primers were also added for the construction of multiplex PCR method.Nine common intestinal normal and pathogenic strains were used to verify the specificity of slpA multiplex PCR for the detection of C.difficile.Forty-six C.difficile reference strains,belonging to 11 slpA genotypes,were used to verify the ability of the multiplex PCR method for dectecting and genotyping.Thirty-nine moxifloxacin-resistant clinical isolates were genotyped by the multiplex PCR,and its clinical value was evaluated by comparing with slpA sequence typing (slpA ST)method.Results All the 9 intestinal normal and pathogenic strains were negative when detected by the multiplex PCR.And tpi of 46 C. difficile reference strains were positive,and 36 strains belonging to slpA genotypes gr,hr,fr,gc08 and 078 were genotyped correctly.Other 10 strains which belonged to other 6 genotypes were non-typeable. Among 39 moxifloxacin-resistant clinical isolates,all were positive of tpi,and 32 isolates were typed correctly by the multiplex PCR method,including 22 slpA genotypes gc08,6 genotypes hr,2 genotypes fr,and 2 genotypes 078,which were consistent with slpA ST.However,7 isolates could not be typed by multiplex PCR,which were identified as other genotypes not included in the multiplex PCR by slpA ST. Conclusions A convenient and rapid multiplex PCR method for the detection of C.difficile is established successfully,which can distinguish among five slpA genotypes.slpA genotype gc08 is the common genotype of moxifloxacin-resistant clinical isolates.

PCR-ribotyping for genotyping Clostridium dif ficile clinical isolates / 国际检验医学杂志

Objective To investigate the genotype and production of toxin A and B of C .difficile clinical isolates collected from Sydney ,Australia .Methods Sixty‐eight C .difficile clinical isolates were collected from Westmead Hospital ,the University of Sydney ,which were genotyped by using PCR‐ribotyping ,and toxin A ,B coding gene tcdA ,tcdB were detected by using PCR meth‐od .Results Thirty‐one PCR‐ribotypes (RTs) were confirmed in the 68 C .difficile clinical isolates ,RT014 (19 .1% ) and RT002 (11 .8% ) were the common genotypes .Sixty‐four of 68 (94 .1% ) isolates contained tcdA and tcdB for toxin A and B .Conclusion The common prevalent PCR‐ribotypes of C .difficile were RT014 and RT002 in Sydney ,most of the C .difficile clinical isolates contained toxin A and B .

Multilocus sequence typing indicates diverse origins of invasive Candida tropicalis isolates in China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>According to data from the China Hospital Invasive Fungal Surveillance Net (CHIF-NET) 2010, Candida tropicalis (C. tropicalis) is the third most common pathogen causing invasive candidiasis. Moreover, the majority of fluconazole-resistant C. tropicalis isolates were from a single hospital. Therefore, a molecular epidemiological survey is necessary to investigate the genetic relatedness of C. tropicalis isolates in China.</p><p><b>METHODS</b>In this study, 48 C. tropicalis isolates causing invasive fungal infections from four tertiary hospitals in China were studied. All the isolates were identified by sequencing the internal transcribed spacer region. Antifungal susceptibility to triazoles, amphotericin B, and caspofungin was determined by the Clinical and Laboratory Standards Institute standard broth microdilution method. Multilocus sequence typing (MLST) was performed, and phylogenetic analysis was further performed by the eBURST and maximum parsimony (MP) methods to characterize the genetic relatedness of isolates.</p><p><b>RESULTS</b>MLST discriminated 40 diploid sequence types (DSTs) among 48 isolates, including 36 novel DSTs, and the XYR1 gene showed the highest discriminatory power. The DSTs obtained from this study were compared with those of previously reported C. tropicalis isolates, and there was poor type alignment with regional strains. Nine groups and 11 singletons were identified by eBURST, whereas two groups and 10 subgroups were clustered by MP analysis. Generally, there were no obvious correlations between clonal clusters generated and the specimen source or hospital origin. Seven fluconazole-resistant isolates were confirmed and assigned to three distinguishable branches.</p><p><b>CONCLUSIONS</b>The results suggested diverse origins of invasive C. tropicalis isolates in China. Although most invasive C. tropicalis strains in the mainland of China were clustered with previously characterized Asian isolates, major C. tropicalis clusters identified in this study were genetically distinct from those of other geographic regions.</p>

Detection of TAC1 gene point mutations in fluconazole-resistant Candida albicans isolates with rolling circle amplification / 中华皮肤科杂志

Objective To detect point mutations of TAC1 gene in fluconazole-resistant Candida albi-cans isolates with rolling circle amplification (RCA), develop an accurate, rapid and specific assay to detect single nucleotide polymorphisms (SNPs), and to estimate the relationship between mutations of TAC1 gene and resistance to fluconazole. Methods A total of 33 fluconazole-resistant Candida albicana isolates, including 8 strains from America and 25 from Australia, were collected. Four TAC1-specific padlock probes were designed according to previously reported mutations. DNA was extracted from these tested strains, subjected to amplification of three targeted fragments of TAC1 gene with PCR. Then, RCA was performed to detect point mutations of TAC1 gene in resistant Candida albicans strains. At the same time, the target fragments underwent sequencing analysis, and the results of RCA were compared with those of sequencing. Results Two types of resistance-associated mutations were found in 5 out of the 33 fluconazole-resistant strains. Among the 5 strains, 4 were from America, 1 harbored T225A mutation and 4 carried A736V mutation. No related mutation was found in TAC1 gene of 4 fluconazole-sensitive isolates. Conclusions RCA assay could accurately and rapidly detect point mutations of genes. Further studies are required to clarify the relationship between TAC1 point mutations and fluconazole resistance.

Detection of five common cell culture contaminant Molllcutes using rolling circle amplification / 中华微生物学和免疫学杂志

Objective To develop a novel padlock probe and rolling circle amplification(RCA) to detect 5 common cell culture contaminant Mollicute( Mycoplasma arginini, Mycoplasma fermentans, Mycoplasma hyorhinis, Mycomplasma orale, and Acholeplasma laidlawii ). Methods "Universal" primers ( SPS1, SPA2 and SPS2, SPA1) were used to amplify the Mollicute 16S-23S rRNA intergenic spacer region.Amplicon was ligate Mollicute specific padlock probe. Probe amplified and monitored using a Corbett RotorGeneTM 6000 machine. Results Five reference Mollicute strains were correctly detected by RCA method.There was no cross-reaction. RCA method can sensitive detect 10 copies templates and show strong positive signal. Sixty-two cell culture specimens were detected. Thirty-seven samples were contained single specie Mollicute and 14 samples contained two Mollicutes. Eleven samples did not contained Mollicute. RCA detection results were concordant with previously species-specific PCR. Conclusion RCA can rapid, sensitive and specific detect contamination Mollicute.

Development of a multiple PCR-based reverse line blot hybridization assay(mPCR-RLB)to detect seven sexually transmitted disease pathogens / 中华皮肤科杂志

Objective To develop a multiple PCR-based reverse line blot hydrization assay (mPCR-RLB)to simutaneously detect several STD pathogens:Neisserria gonorrhoeae,Chlamydia trachomatis,Ureaplasma urealyticum,U.parvum,Mycoplasma genitalium,M,hominis and Trichomonas vaginalis.Methods Seven pairs of biotin-labelled primer were designed and synthesized to target the 16S rRNA-23S rRNA intergenic spacer regions of Neisserria gonorrhoeae,Chlamydia trachomatis,Mycoplasma,and repetitive DNA sequence of Trichomonas to identify and subtype thesc pathogens.DNA was extracted from the referrence strains of seven pathogens and used as templates.mPCR was performed to simutaneously amplify the target regions of these pathogens.Then,the biotin-labelled amplicons were hybridized with membrane-bound specific oligonucleotide probes followed by the detection of bound amplicons with chemiluminescence assay.Serially diluted plasmids containing the target genes of pathogens were amplified with this method to detect its sensitivity.Two-hundred and eleven specimens,including 104 male urethral swabs and 107 female cervical swabs,were collected from the STD clinic of Wuhan First Hospital;mPCR-RLB and single-primer PCR were performed.For specimens with inconsistent results,nested PCR was performed to confirm the results.Results The assay sensitively and specifically identified referrence strains of the tested pathogens.The detection limit of mPCR-RLB was 100 copies for all the pathogens.Of the 211 clinical specimens,2.8%(6/21)were negative for single-primer PCR,but positive for mPCR-RLB,and nested-PCR results were consistent with those of mPCR-RLB.Conclusion mPCR-RLB is a sensitive,specific and rapid method for the detection of STD pathogens from clinical specimens.

Effect of Rehabilitation Evaluation and Treatment on Guillain-Barré Syndrome / 中国康复理论与实践

@#ObjectiveTo explore the effect of rehabilitation evaluation and treatment on Guillain-Barré syndrome.MethodsThe rehabilitation treatment was performed in 21 patients with Guillain-Barré syndrome, and the therapeutic effect was evaluated with Lindmark Assessment.ResultsThe total effective rate of all patients was 90.47% in the upper limps and wrists, 85.71% in the hands and 95.23% in the lower limps.ConclusionThe rehabilitation can improve the therapeutic effect of Guillain-Barré syndrome.

The Application of Problem-based Teaching Method to the Clinical Teaching of Rehabilitation Medicine / 中华医学教育探索杂志

0.05).But,there's a remarkable difference in the learning interests,the abilities for self-taught,analysis and induce abilities,clinical thought and the comprehensive abilities for the two methods(P

Detection of Metallo-?-lactamase and Antibiotics Resistance of Imipenem-resistant Pseudomonas aeruginosa in Qingdao Area / 中华医院感染学杂志

OBJECTIVE To find out the imipenem-resistan metallo-?-lactamase in P.aeruginosa to provide the proof of treatment for clinic.METHODS A multi-disk was used to detect the metallo-?-lactamase of P.aeruginosa and the K-B disk method was used for monitoring of the antibiotic-resistance to 15 antibiotics.RESULTS Fifteen strains producing metallo-?-lactamase were isolated from 93 imipenem-resistant P.aeruginosa strains.The positive rate with metallo-?-lactamase was 16.1%(15/93).The susceptibility tests showed the lower resistance was to cefoperazone sulbactam(Sulperazone),amikacin and piperacillinl tazobactam(Tazocin).their resistance rate respectively was 16.8%,24.8% and 26.6%.The resistance rate to ciprofloxacin and ceftazidime was 43.7% and 33.6%.The resistance rate over 90.0% was to ampicillin,ampicilillin/sulbactam,cefazolin,cefpodoxime,cefuroxime and so on.CONCLUSIONS P.aeruginosa with imipenem-resistance shows seriou multidrug-resistance.The metallo-?-lactamase is the main reason for P.aeruginosa resistance to imipenem and cephalosporins.The doctors should choose antibiotics reasonably according to the susceptibility test when taking effective treatment to P.aeruginosa infection.