Child ; Child, Preschool ; Humans ; Macrophage Activation Syndrome ; etiology ; pathology ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; pathology

Objective To observe the clinical efficacy and adverse reactions of tigecycline in treatment of health-care-associated pneumonia (HAP ) caused by extensively drug-resistant Acinetobacter baumannii (XDRAB ). Methods Clinical data of patients who used tigecycline for the treatment of XDRAB HAP in intensive care units of a hospital from March 2013 to June 2014 were retrospectively analyzed.Results XDRAB isolated from 31 patients with HAP were all sensitive to tigecycline,the resistance rates to carbapenems and sulbactams (including cefopera-zone-sulbactam,SCF)were all 100%,17 cases (54.84%)were mixed infection.Combined use rates of tigecycline and SCF were 85.71 %(12/14)in respiratory intensive care unit(RICU)and 47.06%(8/17)in general intensive care unit(GICU).Of 31 patients,the cure rate,effective rate,bacterial clearance rate,and antimicrobial adverse reac-tion rate were 29.03%,45.16%,61 .29%,and 16.13% respectively,no serious adverse drug reactions occurred. In RICU group and GICU group,the cure rates were 42.86% and 17.65% respectively,effective rates were 71 .43% and 23.53% respectively,and bacterial clearance rates were 78.57 % and 47.06% respectively,difference in effective rate between two groups was significant (P <0.05).Among patients receiving combination of tigecycline and SCF as well as not receiving combined SCF,the cure rates were 35.00% and 18.18% respectively,effective rates were 60.00% and 18.18% respectively,and bacterial clearance rates were 65.00% and 54.55% respectively, difference in effective rate between two groups was significant (P <0.05).Conclusion Tigecycline has a good clini-cal efficacy and little adverse reaction in treating XDRAB HAP;tigecycline combined with SCF is a good choice.

Child ; Child, Preschool ; Complement C3 ; Disease Progression ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; Lipid Metabolism ; Male ; Obesity ; blood

Objective: To determine the noninvasive imaging efficacy of 99mTc-sandostatin scintigraphy for lung cancer. Methods: 57 consecutive patients with pulmonary nodules(PN) were studied with 99mTc-sandostatin scintigraphy.Planar imaging was obtained after injection of (991.6?187.59) MBq of 99mTc-sandostatin at 1.5-4 hour with GE Dual-head gamma camera(Millennium VG, Hawkeye ;General Electric Medical Systems) . SPECT images of the chest were performed at 4-6 h post injection. All scintigraphically detected lesions were confirmed by histopathological analysis and/or by other imaging modalities.Tumor to normal tissues ratios (T/N) were calculated . Results: Out of 57 patients ,47 were malignant tumors; 12 with small cell lung cancer (SCLC), 35 with non- small cell lung cancer (NSCLC). 10 had benign lesions. The sensitivity , specificity and accuracy of 99mTc-Sandostatin in detection of lung cancer were 95.7%, 90%, 94.7%, respectively. In two patients (pts) with pulmonary squamous cell cancer 99mTc-Sandostatin imaging was false negative. In 10 pts. with benign PN, 9 pts. was true negative, but one patient with tubercolama was false positive. T/N ratio was 3.43?0.66, 2.24?0.31 in SCLC and NSCLC respectively. The T/N ratio was higher in small cell lung cancer than NSCLC(t = 4.072 ,P

To examine the role of ultrasound in gene delivery in vitro, three cells lines were exposed to the low-frequency ultrasound of varying intensities and for different durations to evaluate their effect on gene transfection and cell viability of the cells. Microbubble (MB), Optison (10%), was also used to observe the role of the microbubbles in gene transfection. The results demonstrated that as the ultrasound intensity and the exposure time increased, the gene transfer rate increased and the cell viability decreased, but at high energy intensities, the cell viability decreased dramatically, which caused the transfer rate to decrease. The most efficient ultrasound intensity for inducing gene transfer was 1 W/cm(2) with duration being 20 s. At the same energy intensity, higher ultrasound intensity could achieve maximal gene transfer rate earlier. Microbubbles could increase ultrasound-induced cell gene transfer rate by about 2 to 3 times mainly at lower energy intensities. Moreover, microbubbles could raise the maximum gene transfer rate mediated by ultrasound. It is concluded that the low-frequency ultrasound can induce cell gene transfer and the cell gene transfer rate and viability are correlated with not only the ultrasound energy intensity but also the ultrasound intensity, the higher ultrasound intensity achieves its maximal transfer rate more quickly and the ultrasound intensity that can induce optimal gene transfer is 1 W/cm(2) with duration being 20 s, and microbubbles can significantly increase the maximal gene transfer rate in vitro.
3T3 Cells ; CHO Cells ; Cell Line ; Cell Survival/*genetics ; Contrast Media/metabolism ; Cricetinae ; Cricetulus ; Microbubbles ; Transfection/*methods ; Ultrasonics

Objective To evaluate the value of CK34?E12,p63 and P504S immunostaining in diag- nosis of benign and malignant lesions of the prostate.Methods Expression of CK34?E12,p63 and P504S in 74 benign and malignant lesions of prostate,including 27 PC,6 HGPIN,10 LGPIN,3 AAH and 28 BPH were observed by immunohistochemical method.Results Expression of p63 and CK34?E12 were positive in AAH,LGPIN and BPH,all of PC were negative,the positive rate of HGPIN was 83.3%(5/6).There were sig- nificant differences in p63 and CK34?E12 expression between PC and AAH,HGPIN,LGPIN,BPH(P

Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Child ; Female ; Gene Expression Regulation, Neoplastic ; Hodgkin Disease ; genetics ; metabolism ; Humans ; Ki-1 Antigen ; metabolism ; Male ; Middle Aged ; PAX5 Transcription Factor ; metabolism ; Staining and Labeling ; methods ; Young Adult

Objective To explore the effects of P85,microbubbles and ultrasound on plasmid DNA skeletal muscle gene transduction of mice in vivo. Methods Plasmid encoding green fluorescent protein (GFP) ,which conjugated with 0.05% P85 and/or microbubbles, 10% Optison,was injected into the tibialis anterior(TA) muscle of mice with or without ultrasound irradiation (1 MHz, 1 W/cm2 2 min,20% duty cycle). Mice were killed 1 week after injection. The TA muscles were removed and snap-frozen immediately in isopentane cooled by liquid nitrogen and sections 7 μm thick were cut at intervals. One set of sections mounted with DAPI were used to assess the transfection efficiency by counting the number of GFP-positive fibers under fluorescence microscopy,and the other set of sections were stained with haematoxylin and eosin to assess the tissue damage area. Results The P85 and Optison significantly enhanced the plasmid DNA skeletal muscle gene delivery in vivo separately (P<0.01, P<0.05).Ultrasound exposure could significantly enhance the efficiency of P85 induced gene delivery(P<0.01) but not of Option(P>0.05).The gene delivery efficiency induced by P85 was higher than that by Optison no matter with or without ultrasound irradiation(P<0.01). When the P85 conjugated with Optison, they could further significantly enhance gene delivery efficiency with ultrasound exposure (P<0.01). Meanwhile, ultrasound exposure could increase the muscle damage areas in the groups with microbubbles (P<0.01). Conclusions The P85,microbubbles and ultrasound exposure display synergistic effect to enhance plasmid DNA transduction in skeletal muscle of mice in vivo.

OBJECTIVEPrader-Willi syndrome (PWS) is an example of a human genetic disorder that involves imprinting genes on the proximal long arm of chromosome 15 and SNRPN gene as a candidate gene for this syndrome. The purpose of this study was to show the molecular genetic defects and genomic imprinting basis in Chinese PWS patients and to evaluate the clinical applications of a differential diagnostic test for PWS.

METHODSFluorescence in situ hybridization (FISH) and methylation-specific PCR (MSPCR) techniques were applied for 4 clinically suspected PWS patients. Using three probes, including SNRPN probe for identification of the critical locus in PWS region, D15Z1 and PML control probes for identification of the 15p arm and 15q arm, the authors detected the deletions 15q in PWS. MSPCR was based on sodium bisulfite treatment of DNA and PCR primers specific for the maternal and paternal allele.

RESULTSWhen hybridized with mixed probes, it was found in 2 patients that the central specific signal was absent, but both the flanking control signals were retained, indicating SNRPN gene deletion of chromosome 15q11-13. Bisulfite-modified DNA from all PWS children amplified with methylated allele-specific primer pair showed only maternal 131bp PCR product, indicating the maternal uniparental disomy (UPD15).

CONCLUSIONGenomic imprinting plays an important role in the molecular pathogenesis of PWS that caused by paternal microdeletions of 15q11-q13 or maternal UPD of chromosome 15. The basic defect seemed to be an absence of function of PWS genes that are normally expressed only from the paternal chromosome 15. MSPCR is a rapid and simple PCR-based assay compared with other cyto-molecular tests and its results were consistent with the clinical diagnosis of PWS, so it seems to be a reliable diagnostic method for PWS patients who show abnormal methylation at SNRPN. The genetic differential tests for PWS are important in determining familial recurrence risk.

Adolescent ; Autoantigens ; Chromosome Deletion ; Chromosomes, Human, Pair 15 ; genetics ; Gene Deletion ; Genomic Imprinting ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Polymerase Chain Reaction ; methods ; Prader-Willi Syndrome ; genetics ; Ribonucleoproteins, Small Nuclear ; genetics ; snRNP Core Proteins

OBJECTIVETo construct the recombinant plasmid of RCAS1, to express and purify its fusion protein GST-RCAS1, and to investigate its biological function.

METHODSRCAS1 encoding gene was amplified by RT-PCR from total RNA extract of MCF-7 cells and was ligated with expression plasmid vector pGEX-2T by T4 DNA ligase after digested by the restricted endonucleases BamH I and EcoR I. Then the ligated products were inserted into competence JM109 E. Coli and the positive recombinants were identified by restriction endonuclease digestion assay and DNA sequencing. The GST-RCAS1 fusion protein expression was induced by IPTG in BL21 E. Coli and was purified with GST column and identified by SDS-PAGE and Western blotting with anti-GST monoclonal antibody, anti-RCAS1 (N-18) and anti-RCAS1 (C-20) polyclonal antibody. The apoptosis of activated T cells induced by GST-RCAS1 fusion protein was detected by flow cytometry with Annexin V and propidium iodide (PI) staining.

RESULTA 642 bp product was cloned by RT-PCR and the recombinant plasmid was constructed successfully. The GST-RCAS1 fusion protein was recognized by GST monoclonal antibody and RCAS1(N-18 and C-20) polyclonal antibody. FACS analysis showed that GST-RCAS1 fusion protein induced apoptosis in activated T cells.

CONCLUSIONThe recombinant plasmid of RCAS1 has been successfully constructed and the GST-RCAS1 fusion protein expressed and purified. The apoptosis inducing effect of GST-RCAS1 fusion protein on activated T cells is demonstrated.

Antigens, Neoplasm ; biosynthesis ; genetics ; Base Sequence ; Breast Neoplasms ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification ; Tumor Cells, Cultured