Objective To analyze the clinical characteristics, risk factors and prevention measures of nosocomial infection in senile patients with hematologic disorders. Methods The clinical data of 581 senile patients with hematologic disorders from July 2005 to June 2007 were retrospectively analyzed. Results The nosoeomial infection rate was 30.5%(177/581). The 581cases developed nosocomial infection for 254 times [43.7% (254/581)]. Lower respiratory tract infection was the most frequent infection site, followed by intestinal infection and upper respiratory infection. The species were predominated by Gram-negative bacillus (67.1%) . The risk factors of nosocomial infection were non-remission of malignant diseases,chronic underlying diseases, long time hospitaliation, community infection, granulocytopenia, invasive manipulation and application of adrenocortical hormone and antibiotic. Conclusions The nosoeomial infection rate is high in senile patients with hematologic disorders and it can be decreased by taking prevention measures according to the risk factors.

Objective: To explore the regulative role of extracellular regulated protein kinase-5 (ERK5)/Bcl-2 interacting mediator of cell death (Bim) pathway in hypothermal stimulation induced neonatal rat’s cardiomyocytes (CMs) damage and apoptosis.
Methods: CMs were cultured for hypothermal stimulation and the speciifc siRNA was used to down-regulate the ERK5 or Bim in CMs. The cell apoptosis was detected by lfow cytometry, protein expression was examined by Western blot analysis, the intracellular Ca2+, reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) were evaluated by lfuorescent labeling and lfow cytometry.
Results: In hypothermal stimulated CMs, ERK5 siRNA could promote Bim protein expression, but Bim siRNA could not inlfuence ERK5, while attenuated p-ERK5 expression. ERK5 siRNA induced higher apoptosis rate, while Bim siRNA could decrease such effect. ERK5 siRNA increased the intracellular Ca2+overloading, ROS activation andΔΨm damage, while Bim siRNA played the role to against those effects in hypothermal stimulated CMs.
Conclusion: Our study revealed that ERK5/Bim pathway played the important regulative roll in hypothermal stimulation induced neonatal rat’s CMs damage and apoptosis.

Objective To investigate the clinical efficacy of internal fixation through posterior minimally invasive approach in treating unstable scapula fractures.Methods A retrospective case control study was conducted on the clinical data of 36 patients with scapular fractures admitted between May 2011 and August 2016.There were 30 males and six females,with average age of 51.5 years (range,46-64 years).According to Hardegger classification,there were 18 patients with scapular body fracture,14 with scapular neck fracture,and four with glenoidal fracture.According to operation method,the patients were divided into Group A (n =24) which adopted internal fixation through posterior minimally invasive approach and Group B (n =12) which adopted the conventional Judet approach for internal fixation.The incision length,operation time,intraoperative bleeding,fracture healing time,Hardegger standard of clinical effects,and complications in two groups were compared.Results All patients were followed up for average 18 months (range,6-24 months).The total length of surgical incision was (12.50 ± 4.50) cm in Group A and (27.95 ± 5.20) cm in Group B (P ＜ 0.05);the operation time was (86.5 ± 1 1.5) minutes in Group A and (120.6 ± 10.9) minutes in Group B (P ＜ 0.05);the intraoperative bleeding was (200.0 ± 20.0)ml in Group A and (420.0 ± 20.0)ml in Group B (P ＜ 0.05);fracture healing time was (10.0 ± 1.0) weeks in Group A (12.0 ± 1.5) weeks in Group B (P ＜ 0.05).According to Hardegger standard,in Group A,15 patients were excellent,six good,and two fair,with an excellent and good rate of 91％;while in Group B,six patients were excellent,three good,two fair,and one poor,with an excellent and good rate of 75％ (P ＜ 0.05).No complications were observed in Group A.In Group B,one patient with hematoma and one patient with nonunion of incision area were observed,both of which recovered after drainage and dressing change.One patient with superior scapular nerve injury was found and recovered after treatment.The incidence of complications was 25％ in Group B,higher than 0 in Group A (P ＜ 0.05).There was no internal fixation fracture or nonunion in two groups.Conclusion Compared with the conventional Judet approach,minimally invasive internal fixation approach in the treatment of unstable scapula fracture demonstrates the advantages of mild trauma,faster functional restoration,and fewer complications.

OBJECTIVETo investigate the effects of culture supernatant of human amnion mesenchymal stem cells (hAMSCs-CS) on biological characteristics of human fibroblasts.

METHODS(1) hAMSCs were isolated from deprecated human fresh amnion tissue of placenta and then sub-cultured. The morphology of hAMSCs on culture day 3 and hAMSCs of the third passage were observed with inverted phase contrast microscope. (2) Two batches of hAMSCs of the third passage were obtained, then the expression of vimentin of cells was observed with immunofluorescence method, and the expression of cell surface marker CD90, CD73, CD105, and CD45 was detected by flow cytometer. (3) hAMSCs-CS of the third passage at culture hour 72 were collected, and the content of insulin-like growth factor Ⅰ (IGF-Ⅰ), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF) were detected by enzyme-linked immunosorbent assay. (4) Human fibroblasts were isolated from deprecated human fresh prepuce tissue of circumcision and then sub-cultured. Human fibroblasts of the third passage were used in the following experiments. Cells were divided into blank control group and 10%, 30%, 50%, and 70% hAMSCs-CS groups according to the random number table (the same grouping method below), with 48 wells in each group. Cells in blank control group were cultured with DMEM/F12 medium containing 2% fetal bovine serum (FBS), while cells in the latter 4 groups were cultured with DMEM/F12 medium containing corresponding volume fraction of hAMSCs-CS and 2% FBS. The proliferation activity of cells was detected by cell counting kit 8 and microplate reader at culture hour 12, 24, 48, and 72, respectively, and corresponding volume fraction of hAMSCs-CS which causing the best proliferation activity of human fibroblasts was used in the following experiments. (5) Human fibroblasts were divided into blank control group and 50% hAMSCs-CS group and treated as in (4), with 4 wells in each group, at post scratch hour (PSH) 0 (immediately after scratch), 12, 24, 48, and 72, the migration distance of cells was observed and measured with inverted phase contrast microscope. (6) Human fibroblasts were grouped and treated as in (5), with 3 battles in each group, and apoptosis rate of cells was detected by flow cytometer. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, one-way analysis of variance, LSD test, and t test.

RESULTS(1) On culture day 3, most hAMSCs were in large form, and spindle-shaped with much prominences like fibroblasts or in flat polygonal shape. hAMSCs of the third passage were spindle-shaped. The expression of vimentin of hAMSCs of the third passage was strongly positive, and the expressions of surface markers CD90, CD73, and CD105 of the cells were positive, while the expression of CD45 of the cells was negative. (2) The content of IGF-Ⅰ, VEGF, EGF, and bFGF in hAMSCs-CS were respectively (11.7±1.0), (316±68), (6.1±0.4), and (1.49±0.05) pg/mL. (3) At culture hour 12-72, the proliferation activity of human fibroblasts in each hAMSCs-CS group was significantly higher than that in blank control group (with P values below 0.01), and the proliferation activity of human fibroblasts in 50% hAMSCs-CS group was the highest. (4) The width of scratch in two groups was nearly the same at PSH 0. The migration distance of cells in 50% hAMSCs-CS group was significantly longer than that in blank control group at PSH 12-72 (with P values below 0.01). (5) The apoptosis rate of human fibroblasts in blank control group was (16.2±2.4)%, which was significantly higher than that in 50% hAMSCs-CS group [(7.4±3.6)%, t=6.710, P<0.01].

CONCLUSIONShAMSCs-CS can promote proliferation and migration of human fibroblasts and inhibit the apoptosis of human fibroblasts.

Amnion ; cytology ; Apoptosis ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Culture Media, Conditioned ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Epidermal Growth Factor ; metabolism ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Fibroblasts ; cytology ; drug effects ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Male ; Mesenchymal Stromal Cells ; chemistry ; Pregnancy ; Vascular Endothelial Growth Factor A ; metabolism

UNLABELLEDWe designed a pair of specific primers and a TaqMan fluorescent probe targeting the toxR gene of Vibrio parahaemolyticus (VP). After optimizing the conditions, the specialty, sensitivity and reproducibility of the detection method were evaluated.

RESULTS(1) the developed real-time PCR assay protocol detected only VP and was not affected by other normal food pathogens such as Staphylococcus aureus, Salmonela, Listeria monocytogenes. (2) the limit of detection was 25 copies of toxR gene in the detected samples, and the sensitivity of pure cultures and simulated food samples was 21 cfu/mL and 210 cfu/g. (3) the developed protocol of real-time PCR assay had a high reproducibility, and the sample's variation was 0.9% and 1.3% within the same sample and between tests. (4) the standard curve had a good linearity when the gene quantity was between 2.5x10(1) and 2.5x10(6) copies. The developed detection assay targeting the toxR gene can quantitatively detect VP in only 3 hours, and thus is an efficacious method for the detection of Vibrio parahaemolyticus.

Bacterial Proteins ; genetics ; DNA-Binding Proteins ; genetics ; Fluorescent Dyes ; metabolism ; Gene Targeting ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Taq Polymerase ; metabolism ; Transcription Factors ; genetics ; Vibrio parahaemolyticus ; genetics ; isolation & purification

We formulated a selective enrichment broth (SVV) for simultaneous growth of Salmonella, Vibrio parahaemolyticus, and Vibrio cholerae by single factor experiment and response surface method. We evaluated the enrichment effect of SVV by conventional culture method and real-time PCR assay. We obtained the SVV broth by supplementting the Buffered Peptone Water (BPW) with bile salt no. 3, potassium tellurite, and sodium citrate as inhibitors, and glucose, mannitol, snhydrous sodium sulfite and sodium pyruvate as accelerants. We also modified the concentration of sodium chloride in BPW. When mixed at equal or varied proportions, the target pathogens had a great accumulation (10(5)-10(8) CFU/mL) after incubated in SVV for 18 h at 37 degrees C with shaking. It can also effectively inhibit the competitive microflora. We detected 10 artificial simulated samples and 608 real samples using SVV with real-time PCR. After enriched in SVV for 18 h, the quantity of the bacteria in samples were above the detection limit. The SVV with PCR assay showed higher tested positive (4.06%) compared to that of the conventional detection method (3.78%) and there was no false report. In summary, SVV is a promising new multiplex selective enrichment broth that can be used in detection of seafood.
Culture Media ; Food Microbiology ; Salmonella ; growth & development ; Vibrio cholerae ; growth & development ; Vibrio parahaemolyticus ; growth & development

Pseudomonas aernginosa (PA) is one of the most universal pathogens in clinical diagnosis, and conventional detection assay has many disadvantages. In this research, a pair of specific primers and a TaqMan fluorescent probe were designed in the conservative region of ETA gene by the method of bioinformatics analysis, the detection method for PA was successfully developed. Different gradient concentrations of PA DNA and various pathogen DNA were amplified by fluorescence quantitative PCR (FQ-PCR) to confirm the specificity and sensitivity of the developed method. Results showed that the developed detection assay is more sensible and specific by comparison to the conventional FQ-PCR method, and it is valuable for research and application prospects.
ADP Ribose Transferases ; genetics ; Bacterial Toxins ; genetics ; DNA, Bacterial ; analysis ; Exotoxins ; genetics ; Fluorescent Dyes ; Fluorometry ; methods ; Polymerase Chain Reaction ; methods ; Pseudomonas aeruginosa ; genetics ; isolation & purification ; Sensitivity and Specificity ; Taq Polymerase ; Virulence Factors ; genetics

Objective:To study the effects of cryptotanshinone on proliferation and apoptosis of human bladder carcinoma J82 cells in vitro and its mechanisms.Methods:The J82 cells were treated with cryptotanshinone of different concentrations including 0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 and 8.0 μmol/L.After 48-hour treatment, the proliferation of J82 cells was determined by CCK-8 assay method.Flow cytometric analysis with Annexin V-FITC/PI staining was used to evaluate the apoptosis of J82 cells, and Western blotting was adopted to observe the protein expressions of p65, caspase-3, caspase-8 and caspase-9 related to proliferation and apoptosis.Results:CCK-8 results showed that the A values of 450nm-wavelength were as following: the control group (1.77±0.06), 0.5μmol/L group (1.78±0.08), 1.0μmol/L group (1.64±0.05), 2.0μmol/L group (1.48±0.12), 3.0μmol/L group (1.20±0.07), 4.0μmol/L group (0.93±0.10), 5.0μmol/L group (0.76±0.02), 8.0μmol/L group (0.05±0.01), and the A values were significantly different among the three groups ( F=329.83, P=0.00), there were statistically significant differences between any two groups except the 0.5 μmol/L group(all P<0.05). The early and total apoptosis rates were both significantly different among the three groups ( F=32.49, P=0.00; F=6.39; P=0.03), the early apoptosis rates of 3.0, 1.0μmol/L group were higher than that of the control group(all P<0.05), and the early apoptosis rate in the 3.0 μmol/L group was higher than that in the 1.0μmol/L group[(11.83±1.12)% vs.(7.01±1.84)%, t=3.73, P<0.05]. The expression of p65 protein decreased, while both the expressions of caspase-3 and caspase-9 proteins increased after treatment with cryptotanshinone. Conclusion:Cryptotanshinone can significantly inhibit proliferation and induce apoptosis of human bladder carcinoma J82 cells in vitro, probably via suppressing NF-κB signal pathway and activating mitochondrial pathway, respectively.

Objective To investigate the value of high frequency magnetoencephalography signals in the localization of refractory temporal lobe epilepsy. Methods Retrospective analysis was performed in 10 patients with refractory temporal lobe epilepsy admitted to and accepted surgery in our hospital from January 2015 to December 2015. Surgical approaches of these patients were determined according to the results of long-term video EEG monitoring (VEEG), MR imaging, and conventional and high-frequency magnetoencephalography (MEG). MEG positioning analysis was performed after the surgery; followed up for 12 months was performed to evaluate the surgical efficacies. Results The surgery was effective in all the 10 patients; 5 patients achieved Engel grading Ⅰ, 2 patients achieved Engel grading Ⅱ, and 3 patients achieved Engel grading Ⅲ. The results of high-frequency MEG analysis indicated that 8 lesions were consistent with the surgical sites, enjoying good results; while the positioning error of the 2 patients was large. Conclusions The localization analysis of high-frequency neuromagnetic signals has the potential to determine epileptogenic zones preoperatively for epilepsy surgery. High-frequency oscillation is a new biomarker for the diagnosis of epilepsy.