Objective To establish a flow cytometry-based drug susceptibility test for the rapid de-tection of antifungal susceptibility or resistance of Candida isolates.Methods The gate selection and opti-mal experimental conditions of flow cytometry-based drug susceptibility test were determined by using Candi-da albicans strain ATCC90029 as the test strain and propidium iodide ( PI) as the fluorescent dye .The es-tablished flow cytometry-based drug susceptibility test was used to detect the susceptibility or resistance to fluconazole or voriconazole of 110 isolates belonging to Candida species, and the obtained results were com-pared with those by using typical M 27-A3 constant dilution method .Results The killed and viable Candida albicans ATCC90029 strains were clearly divided into two groups on the figure of SS /log (FL3) by regulating voltages.There was a high correlation between the results of susceptibility test and the proportions of killed and viable fungi in mixture (r=0.999).The flow cytometry-based drug susceptibility test could provide the results within 30 min and its optimal concentration of fungal suspension , time of drug-fungus incubation , dyeing method and time were 1.0×106/ml, 3 h incubation and sodium deoxycholate-pretreated plus PI dye-ing for 5 min, respectively .The total coincident rates between the established test and the constant dilution method were 98.2%and 87.3%in the detection of drug susceptibility of 110 fungal isolates to fluconazole and voriconazole .Conclusion The flow cytometry-based drug susceptibility test shows advantages of rapidi-ty, accuracy and high sensitivity compared with the constant dilution method .It has a great potential for clin-ical application .

Objective:To supply a reference measure,CD8+CD38+ expressin,AST,TBIL,PT were examined in patients with chronic hepatitis B.Methods:CD8+CD38+ expression of peripheral blood lymphocyte in 38 patients with chromic hepatitis B and 14 patients with liver cirrhosis were detected with flow cytometry. AST,TBIL,PT were also measured at the same time with instruments.Results:(1)CD8+CD38+ expression in patients with choronic hepatitis B and liver cirrcosis elevated significantly compared with those in normal controls. But CD8+CD38+ expression in patients with liver cirrhosis was lower significantly than those with chronic hepatitis B.(2)CD8+CD38+ expression in moderate chronic hepatitis B were higher than those in mild chronic hepatitis B. CD8+CD38+,CD8+,CD38+ expression in severe chronic hepatitis B were higher than those in mild chronic hepatitis B. CD38+ expression in severe chronic hepatitis B was higher than those in moderate chronic hepatitis B. CD8+CD38+,CD8+,CD38+ expression in liver cirrhosis was lower than those in mild,moderate,severe chronic hepatitis B. CD8+ expression in liver cirrhosis was lower than those in severe chronic hepatitis B.(3)CD8+CD38+ expression in patients under AST,TBIL,PT abnormal condition were higher than those in patients under AST,TBIL PT,normal condition.Conclusion:The increasing of CD8+CD38+ expression of peripheral blood lymphocyte in patients with chronic hepstitis B,reflect the fact that cell immunolgy was abnormally activated.There was a remarkable correlation on CD8+CD38+ and AST,TBIL,PT. To test expression of CD8+CD38+ of peripheral blood lymphocyte in chronic hepatitis B has very important clinical reference value on patient′s condition development and diagnosis.

The architecture of the specialized myocardial fibers of the atrioventricular junction area(AVJ) were observed under microscope on serial sections. It was found that the human A-V node extend backward divergently to form two posterior extensions (left and right, LPE, RPE). The A-V node can be divided into a superficial layer and a deep layer, and the deep layer further divided into a upper part and a lower part. The fibers in the superficial layer are vertical, and join with the lower part of the deep layer at the anterior extremity of the node. The lower part of the deep layer is the main axis of the node. The fibers in the lower part of the deep layer are longitudinal, and join with the RPE at the posterior extremity of the node. The fibers in the upper part of the deep layer are oblique and join with the LPE at the posterior extremity of the node, and mix with the lower part at the anterior extremity. The longitudinal dissociation of the anterior part of the His bundle is more distinct than that of the posterior part. The lower-left part of the bifurcating part of the bundle extend to form the posterior left branch, the upper-left and upper-middle parts of the bifurcating part extend to form the anterior left branch, and the right part and lower-middle part of the bifurcating part extend to form the right bundle branch. The morphological evidence of the dual or multiple conducting pathways, the longitudinal dissociation and the reentry pathways in the AVJ were discussed.

OBJECTIVE To analyze the homology of carbapenem-resistant Klebsiella pneumoniae(KPN),offering help for clinical therapy and nosocomial infection control.METHODS The antimicrobial-resistant phenotype of forty carbapenem-resistant K.pneumoniae strains was analyzed by the WHONET 5.4 soft and the resistant genotypes were determined by plasmid profile analysis and pulse-field gel electrophoresis(PFGE).RESULTS Analyzing antimicrobial-resistant phenotype to usual eighteen clinical drugs,the main drug resistant profiles were pan-resistant and only sensitive to tobramycin among the eight antimicrobial-resistant profiles(72.5%).Additionally,the main strains were type Ⅰ and type Ⅱ among the five strains analyzed by plasmid profile(82.5%).When analyzed by PFGE,five types were identified and among these strains type Ⅰ was predominant in 34 strains(85.0%).CONCLUSIONS The strains used in this study exhibit higher homology.Therefore,clinical departments and nosocomial infection departments should pay more attention to these strains to avoid outbreak.

We isolated and identified the bacterial pathogen in a pyogenic balanoposthitis patient and investigated the drug resistance and its mechanism of the isolate.Urethral secretions and balanus pustule liquids were collected for microscopic examination after Gram-staining and detection of mycoplasma using Mycoplasma IST 2 kit.The two samples were inoculated on Columbia blood plate,N.gonorrhoeae selective plate and chromID Candida plate for isolation.The obtained colonies were identified by VITEK 2-compact automatic bacterial detection and analysis system.Moreover,PCR was performed to detect 16S rRNA gene of N.gonorrhoeae in the samples and colonies.KB method was applied for detecting susceptibility of five common antibiotics against the isolate.The β-lactamase and extended spectrum β-lactamase confirmatory tests were used to investigate the enzyme production of the isolate as well as drug resistance-associated tetM,TEM,mefA and ermF genes in the isolate were detected by PCR.Results showed that all the clinic samples showed negative for mycoplasma.All the isolating cultivation results of urethral secretions were negative while the balanus pustule liquids provided positive isolating cultivation in the blood and selective plates.The VITEK 2-compact system and 16S rRNA-PCR revealed that the isolated strain belongs to N.gonorrhoeae.The isolate can produce β-lactamases and resist to penicillin G,ciprofloxacin and tetracycline.The tetM,TEM,mefA and ermF genes could be found in the isolate's genome.The patient's balanoposthitis is caused by infection of N.gonorrhoeae.The multidrug resistance of Neisseria gonorrhoeae isolate is closely associated with its carried resistant genes.

Objective To study the effects of using 25 kDa linear and branched PEI to transfer plasmid DNA pEGFP -C1 (pDNA ,encoding the enhanced green fluorescent protein reporter gene ) to the basilar membrane of the C57BL/6 mice cochlea in vitro .Methods L -PEI/pDNA and B -PEI/pDNA polyplexes were generated in 0 .1M phosphate buffer solution (PBS) or 5% glucose solution .Polyplexes were characterized by transmission electron mi-croscopy .Agarose gel retardation assay was used to determine the plasmid binding ability of L -PEI and B -PEI . The toxicity was investigated by MTT assay .The transfection was firstly evaluated in 293T cell line ,and then the appropriate amount of PEI and plasmid were applied for cochlear explant transfection of P4 mice pups .Results Un-der the same condition ,B -PEI had better transfection efficiency than L -PEI ,but its toxicity was also higher . When generated in PBS ,the polyplexes had lower toxicity than in glucose solution .L -PEI-pDNA nanoparticles could transfect the spiral limbus fibrocytes ,some spiral ganglion neurons and supporting cells ,but the efficiency was low .Conclusion L -PEI could be used as the non -viral vector for the transfection of the cultured basilar mem-brane of P4 mice pups ,but it should be modified to reach higher efficiency .

Background: Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. Methods: Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. Results: The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. Conclusion The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.

Background: Subclinical hypothyroidism (SCH) is the most common thyroid dysfunction, and its relationship with blood pressure (BP) has been controversial. The aim of the study was to analyze the association between SCH and newly-diagnosed hypertension. Methods: Based on data from the Thyroid disease, Iodine nutrition and Diabetes Epidemiology (TIDE) study, 49,433 euthyroid individuals and 7,719 SCH patients aged ≥18 years were enrolled. Patients with a history of hypertension or thyroid disease were excluded. SCH was determined by manufacturer reference range. Overall hypertension and stage 1 and 2 hypertension were diagnosed according to the guidelines issued by the American College of Cardiology/American Heart Association in 2017. Results: The prevalence of overall hypertension (48.7%), including stage 1 (28.9%) and 2 (19.8%) hypertension, increased significantly in SCH patients compared with euthyroid subjects. With elevated serum thyroid stimulating hormone (TSH) level, the hypertension prevalence also increased significantly from the euthyroid to different SCH subgroups, which was more profound in females or subjects aged <65 years. The age- and sex-specific regression analysis further demonstrated the same trends in the general population and in the 1:1 propensity matched population. Similarly, several BP components (i.e., systolic, diastolic, and mean arterial BP) were positively associated with TSH elevation, and regression analysis also confirmed that all BP components were closely related with SCH in female subjects aged <65 years. Conclusion The prevalence of hypertension increases for patients with SCH. SCH tends to be associated with hypertension and BP components in females younger than 65 years.