OBJECTIVE To approach the effect of empirically applying ?-lactam antibiotics for treatment of hospital-acquired pneumonia on distribution and antibiotic-resistance of pathogenic bacteria.METHODS To investigate 141 patients with hospital-acqired pneumonia in intensive care unit during Jan 2001-Oct 2005,and divide into 3 groups:third generation cephalosporin group;lactamase inhibitor group;and other lactam antibiotics group according to different initial antibacterial strategy,then analyze difference in distribution and antibiotic-resistance of pathogens among each group.RESULTS We acquired 164 strains of pathogens.Comparing with other two groups,the proportion of Gram-positive cocci in lactamase inhibitor group was higher significantly(P

Aim To explore the mechanisms of lovastatin protecting EPCs.Methods EPCs were preincubated with lovastatin or LOX-1 mAb for 24 h and then exposed to oxLDL for 48 h.The abilities of migration,adhesion,and tube structure formation of EPCs were examined.To explore the mechanisms,the level of NO,the expression of eNOS and LOX-1 protein and mRNA were assayed.Results Incubation of EPCs with oxLDL resulted in the impairment of migration,adhesion and tube structure formation.Furthermore,oxLDL caused the decrease of NO generation,the down-regulation of eNOS mRNA and protein expression,the up-regulation of LOX-1 mRNA and protein expression.However,the detrimental effects of oxLDL on EPCs function were attenuated by lovastatin and LOX-1 mAb.Moreover,the effects of oxLDL on NO generation,eNOS and LOX-1 expression were reversed by lovastatin and LOX-1 mAb.Conclusion Lovastatin protects EPCs by the regulation of eNOS and LOX-1 expression.

0.05).Conclusion Breath-hold MRCP utilizing the SSFSE technique with high-quality images can accurately assess the level of obstruction and the causes of the obstruction in patients with obstructive jaundice, without the risks of endoscopic retrograde cholangiopancreatography(ERCP).This MRCP technique should be preferred a reliable and noninvasive imaging modality for the diagnosis of obstructive jaundice.

Objective To establish a new multiplex-PCR assay to improve the detection rate of mutations in the DMD gene in Chinese patients. Methods A retrospective review of DMD deletion spectrum of 355 DMD patients with deletions all over the gene was performed. All deletions were confirmed by " one-step approach" diagnostic procedure and MLPA analysis. The exons with high frequency of mutations were identified to constitute the amplification system and the PCR conditions were optimized. Results Two new multiplex-PCR assays were established. Assay one was used to detect 10 exons including exon 5, 8, 17, 44, 45, 47, 49, 50, 51 and 52 of DMD gene, in two PCR sets. The theoretical detection rate would be 92% (326/355). Assay two was used to detect 5 exons including exon 12, 19, 35, 43 and 54, which could be used to screen additional 5% (17/355) deletion cases. The method was validated in other 22 DMD patients. Multiplex-PCR results were completely identical to the MLPA results in all 22 DMD patients. Conclusions The two multiplex-PCR assays were established based on the analysis of 355 Chinese DMD patients with gene deletions. It is believed that the new approach would be more applicable for deletion detection on the Chinese DMD patients since the DMD cases involved were from the whole country.

Purpose To describe clinicopathological features, diagnosis and differential diagnosis of Castleman′s disease. Methods Retrospective analyses of the clinical data, clinicopathology and immunohistochemistry were conducted in ten cases of Castleman dis-ease and reviewed of literature. Results There were 8 cases of unicenrtic Castleman′s disease and 2 cases of multicentric Castleman′s disease. Pathologically, there were 6 cases of hayline vascular types, one case of plasmatcyic type and 3 cases of mixed type in all Castleman′s disease. Immunohistochemically, all cases were negative for BCL-6 and CD10, and Ki-67 expression was less than or e-qual to 30%. There were 4 cases with complete follow-up data, including one case of intermediate type, 3 cases of hyaline vascular type which were healed by surgical resection without recurrence. Conclusions Castleman′s disease is a rare and lymphoproliferative disorders with unknown cause, it is not easy to diagnose before the operation. Whether immunohistochemical features reflect abnormal immune function or play unknown role in the pathogenesis of Castleman′s disease is also demanded further study.

Objective To investigate activation of the phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(Akt)pathway in the endometrium of women with polycystic ovary syndrome(PCOS)and its role in endometrium hyperplasia and carcinogenesis,and the factors affecting the activation of the PI3K/Akt pathway.Methods From Jan 2007 to Jun 2008,52 patients with PCOS who underwent dilatation and curettage were selected as experimental group matched with 32 non-PCOS patients as control group.Serous hormonal parameters,fasting blood glucose and insulin,body mass index(BMI),and endometrium pathology were measured and evaluated in all patients.The PCOS patients were divided into insulin resistance and non-insulin resistance group according to homeostasis model assessment-insulin resistance index (HOMA-IR).Meanwhile,the PCOS patients were grouped as normal,endometrial hyperplasia and carcinoma depending on outcome of pathology.The expression of Akt and phosphorylated Akt(p-Akt)were determined by western blot.Results(1)The expression of p-Akt was significantly higher in PCOS group [(46±18)％]than that in control[(33 ±9)％,P ＜0.01)].(2)The expression of p-Akt was significantly higher in group of endometrial hyperplasia and carcinoma[(56 ± 19)％]when compared with those in normal endometria group[(31 ± 12)％,P ＜ 0.05]; the expression of p-Akt was significantly higher in group of insulin resistance[(50 ± 19)％]compared with that in non-insulin resistance group [(34 ± 10)％,P ＜0.01].(3)There was a positive correlation between the expression level of p-Akt in endometrium with PCOS and HOMA-IR and BMI respectively(r =0.400,0.326,both P ＜ 0.05).Conclusions The PI3K/Akt pathway was over activated in endometrium with PCOS which may be associated with the formation of endometrial hyperplasia and carcinoma in PCOS patients.Insulin resistance and obesity may be high risk factors for over-activation of the PI3K/Akt pathway in endometrium with PCOS.

Auxotrophic strains of N1-37 (Phe-) and N2-27 (His-), screened from mutations of Paenibacillus polymyxa JSa-9 previously, were used as the parent strains to screen high-producing LI-F antibacterial lipopeptide fusion strain through protoplast fusion with polyethylene glycol as a promote agent. Fusion strain F5-15 was obtained. Then the product of LI-F antibacterial lipopeptide was quantified by HPLC, and the difference of expression of the key genes of lipopeptide synthase between wild strain JSa-9 and the fusion strain was analyzed by real-time PCR. LI-F antibacterial lipopeptide yield of the fusion strain F5-15 was 3.1-fold of the original strain JSa9's, and the expression levels of the target genes were 10.48, 2.48, 2.1 and 11.8 fold of the initial strain JSa-9, respectively.
Anti-Bacterial Agents ; biosynthesis ; Chromatography, High Pressure Liquid ; Lipopeptides ; biosynthesis ; Paenibacillus ; metabolism ; Protoplasts ; metabolism ; Real-Time Polymerase Chain Reaction

BACKGROUND: Imatinib has a significant pro-apoptosis effect on chronic myelogenous leukemia (CML), but there are still some patients being resistant to it. Human umbilical cord mesenchymal stem cells (hUC-MSCs) affect the apoptosis of a variety of hematologic malignancies. However, the impacts of hUC-MSCs on the apoptosis of CML cells induced by imatinib remain unclear.OBJECTIVE: To investigate whether hUC-MSCs have an influence on the apoptosis of K562 cells induced by imatinib and to reveal the possible underlying mechanism.METHODS: K562 cells were cultured with hUC-MSCs or/and imatinib. Cellular apoptosis was measured with Annexin-V and PI staining by flow cytometry analysis. The protein expressions of Bax, Bcl-2, caspase-3, caspase-9 and cleaved-PARP in K562 cells were detected by western blot assay. Pan-caspase inhibitor Z-VAD-FMK was used to block apoptosis in each group, and during this process the effect of caspase apoptosis signaling pathway was detected.RESULTS AND CONCLUSION: The apoptosis of K562 cells was enhanced, when imatinib was combined with hUC-MSCs. Western blot analysis showed that the expression of pro-apoptotic protein Bax was enhenced and the expression of anti-apoptotic protein Bcl-2 was suppressed. Furthermore, the cleaved forms of caspase-9, caspase-3 and PARP in K562 cell were higher in the hUC-MSCs+imatinib group than in the imatinib group. The apoptosis of K562 cells induced by the hUC-MSCs combined with imatinib was significantly inhibited by Z-VAD-FMK. In conclusion, these findings indicate that hUC-MSCs can enhance imatinib-induced apoptosis of K562 cells by activating caspase apoptosis signaling pathway.

BACKGROUND: Mesenchymal stem cells (MSCs) are an important component of the in vivo microenvironment and act on multiple biological behaviors of tumor cells. The potential clinical value of MSCs has become an issue of concern in recent years.OBJECTIVE: To investigate the gene expression profiles of acute promyelocytic leukemia (APL) cell line NB4 treated with umbilical cord-derived MSCs (UC-MSCs) using cDNA microarray.METHODS: In vitro co-culture system was constructed, and then cellular proliferation, apoptosis and differentiation status of NB4 cells treated with UC-MSCs were evaluated. Two cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without UC-MSCs. The probes were labeled with fluorescence dyes individually, hybridized with cDNA microarray, and their fluorescent intensities were scanned. The genes were screened through the analysis of the difference in two gene expression profiles.RESULTS AND CONCLUSION: UC-MSCs promoted the proliferation and differentiation, while reduced the apoptosis of NB4 cells. The analysis of gene expression profiles indicated that after co-culture with UC-MSCs, 530 genes were up-regulated and 53 genes were down-regulated. Accordingly, specific gene function and pathway signaling related were also regulated to some extent. Overall, UC-MSCs influence can major biological behaviors of NB4 cells by changing expression of a large amount of genes, gene-related function and multiple intracellular signaling pathways.

ObjectiveTo observe the effect of indomethacin on the migration of breast cancer cell line MCF-7 in vitro and investigate the mechanism involved.MethodsThe migration of MCF-7 cell line stimulated with or without indomethacin were tested using transwell plates consisting upper and lower chambers separated by Millipore polycarbonate membrance filters with 8 μm pore sizes; the levels of chemokine receptor 4(CXCR4),cyclooxygenase(COX-2),epidermal growth factor receptor(EGFR) and vascular endothelial growth factor(VEGF)expression in MCF-7 cell line were detected by flow cytometry,Real-time PCR and ELISA,respectively.Results Indomethacin decreased the migration ability of MCF-7 cell line significandy.CXCR4 membrane expression was significantly reduced in a time-dose dependent manner,and CXCR4,COX-2 and EGFR mRNA levels were significantly downregulated after indomethacin stimulation.However,exposure to indometahcin had no major effect on VEGF production of cells.ConclusionThe downregulation of CXCR4,COX-2 and EGFR expression might be the primary mechanism involved in the inhibitory effect of indomethacin on the migration of MCF-7 cell line.