Objective To study the clinical effect of personalized intervention on elderly patients with colostomy after Miles operation for rectal carcinoma. Methods 114 cases elderly patients with colonic stoma after Miles operation for rectal cancer from June 2014 to January 2016 were divided into control group and observation group by random number method, 57 cases in each. The control group were treated with routine intervention, while patients in observation group was treated with personalized intervention. the self-care ability score before intervention, the incidence of complications, intervention satisfaction and life quality score were compared between the two groups at the same time. Results The total complication rate in the observation group was significantly lower than that in control group ( 5.25% vs 22.80%) (χ2 = 8.36, P = 0.000); the intervention satisfaction of the observation group was significantly higher than that in control group (91.23% vs 75.44%) (χ2 = 6.60, P = 0.010). After intervention, the self-care ability score and life quality score of observation group were significantly higher than that in control group (P < 0.01). Conclusion The personalized intervention can reduce the complications, significantly improve the patients' life quality score and self-care ability, and effectively alleviate the negative situation, improve the patients intervention satisfaction, with a higher development value, it is worth of clinical promoting.

Objective To detect the transferred vascular endothelial growth factor (VEGF)165 gene expression in rhesus autologous bone marrow mesenchymal stem cells (MSCs), and to explore the functional viability of transgenic MSCs. Methods MSCs from rhesus bone were isolated by Ficoll, which were used to detect the phenotype. After the culturing, the expression vector pcDNA-eGFP-VEGF165 was transfected into bone marrow MSCs. Fluorescence microscope and flow cytometry were used to detect the enhanced green fluorescent protein (eGFP) expression. At the same time, the phenotype in transfected MSCs was also indentified. The VEGF165 expression level was detected by RT-PCR. Results The highly purified MSCs were collected successfully. The transfected MSCs and daughter cells showed expressions of eGFP and VEGF165, which also remained the characteristics of MSCs. Conclusion The VEGF165 gene that is transfected into MSCs can maintain characteristics of MSCs, and stably express foreign genes.

In order to amplify pilA gene and ompC gene of avian pathogenic Escherichia coli (APEC) strain, two pairs of primers were designed according to the GenBank sequences, and a 549 bp pilA gene and a 1104 bp ompC gene were obtained by PCR separately. Sequence analysis indicated that the homology of the nucleotide sequence of AEPC strain to those other reference strains was 98.18% of the pilA gene and 97.28% of the ompC gene. Two expression plasmids pETpilA and pETompC were constructed by inserting pilA gene and ompC gene into the prokaryotic expression vector pET-28a. The two plasmids were transformated into E. coli BL21 separately and two recombinant strains BL21 (pETpilA) and BL21 (pETompC) were obtained. The type 1 fimbraie and the out membrane protein were highly expressed when the recombinant strain BL21 (pETpilA) and BL21 (pETompC) were induced by IPTG Two specific proteins were detected by SDS-PAGE and immunogenicity of the expressed protein was confirmed by Western blotting and ELISA. The expressed fimbraie and OmpC were transformed into vaccine. The protective immune response was proved after the mice were immunized with the two vaccines. The results showed that the recombinant strain BL21 (pETpilA) and BL21 (pETompC) could be as candidate vaccine to provide protective immune response against AEPC infection.
Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; immunology ; metabolism ; Escherichia coli Proteins ; genetics ; immunology ; metabolism ; Escherichia coli Vaccines ; immunology ; Fimbriae Proteins ; genetics ; immunology ; metabolism ; Gene Expression Regulation, Bacterial ; Genes, Bacterial ; Mice ; Porins ; genetics ; immunology ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

Estrogen Receptor (ERalpha) is a member of superfamily of ligand-activated transcription factors which play critical roles in many biological processes. To screen novel modulators of ERalpha for drug development and biological function research, we developed a mammalian one-hybrid-based high-throughput screening model for ERalpha modulator. We cloned the ERalpha LBD gene from the total mRNA of fat tissue by RT-PCR and fused it with the GAL4 DNA binding domain of pBIND-GAL4 plasmid to construct a chimara expression plasmid pBIND-GAL4-Eralpha(LBD). The L02 cells was cotransfected with pBIND-GAL4-ERalpha(LBD) and a GAL4-responsive luciferase reporter plasmid pGL3-GAL4, and following treatment with test compounds for 24 h, the activities of luciferase were detected to evaluate the transactivities of ERalpha modulators. After manner optimizations of transfection conditions, Estradiol, an agonist control, induced the expression of luciferase in a dose-dependent with EC50 of 0.17 micromol/L, the maximum folds of induction was about 28.1. Tamoxifen, an antagonist control, efficiently suppressed the estradiol-mediated luciferase induction with EC50 of 0.10 micromol/L. Using this screening model, we discovered four ERalpha agonists from 2000 natural and synthetic compounds.
3T3-L1 Cells ; Animals ; Chimera ; metabolism ; DNA-Binding Proteins ; biosynthesis ; genetics ; Estrogen Receptor Modulators ; chemistry ; isolation & purification ; Estrogen Receptor alpha ; agonists ; Genes, Reporter ; genetics ; Genistein ; chemistry ; isolation & purification ; HeLa Cells ; Humans ; Luciferases ; genetics ; metabolism ; Mice ; Models, Chemical ; Saccharomyces cerevisiae Proteins ; biosynthesis ; genetics ; Transcription Factors ; biosynthesis ; genetics ; Transfection

Objecfive To find out the suitable therapy for the patient of desmoid tumour in head and neck.Methods Foay-four patients with desmoid tumours of the head and neck treated at Cancer Hospital of Fudan University between 1987 and 2002 were identified from inpatient tumour database.Patients were classified into three groups:operation group(15 cases);operation+radiation group(12 cases);radiation group(17 cases).All patients were prospectively follwed.Clinieopathologic features and treatment modalities were evaluated.Results In the group of operation,four of operation group had recurrences 26.7%(4/15).In the group of radiation,three of radiation group had recurrences 17.6% (3/17).And the recurrence of operation+tradiation group Was 23.5%(4/17).No patient died of their disease.Conclusion For desmoid tumors of the head neck,operation+radiation was recommended,and the benefits of radiation therapy were demonstrated.

Objective To evaluate the efficacy of interventional therapy for complex congenital heart defects patients with un-repaired or postoperative residual lesions.Methods Between March 1998 and April 2009,42 patients (12 females),mean age 6 years (5 months to 30 years) received interventional therapy,17 cases underwent occlusion of major acrto-pulmunary collateral arteries (MAPCAs),15 underwent device closure of residual ventricular or atrial septal shunting,12 underwent balloon angioplasty (n = 10) and stenting (n = 2) for stenosis of the anastomosis of vessels or branched pulmonary arteries.Results Twenty-three MAPCAs were performed in 17 patients without residual shunting.One patient died of multiple organ failure after intervention therapy and the remaining patients discharged without complication,successful device closure was performed in 15 patients and there was minimal residual shunting in 1 patient.There were no severe arrhythmias such as complete atrio-ventricular block during and post procedure.Exercise capacities were significantly improved in 12 patients underwent balloon angioplasty or stenting.Pressure gradients were significantly decreased and there was no aneurysmal or thromboembolic formation post procedure.Conclusions Interventional therapy is a safe and effective therapy option for treating complex congenital heart defects patients with un-repaired or postoperative residual lesions.

The present study aims to explore the possible mechanisms that trichostatin A (TSA), a histone deacetylases inhibitor (HDACi), affects the inflammatory signaling pathways of lipopolysaccharide/toll-like receptor 4/nuclear factor-κB (LPS/TLR4/NF-κB). Murine macrophage cell line RAW264.7 cells were employed. MTT assay was used to assess cell viability. The contents of TNF-α, IL-1β and IL-6 in culture supernatant were assayed by enzyme-linked immunosorbent assay (ELISA). TLR4 expression and NF-κB/p65 (Lys310) acetylation were examined by Western blotting. DNA binding activity of NF-κB/p65 was detected by using TransAM(TM) NF-κB/p65 activity assay kit. The results showed that, compared with control group, which was treated by DMSO, the cells treated with TSA (20, 40, 80 ng/mL) showed decreased percentages of cell survival (P < 0.05). The contents of TNF-α, IL-1β and IL-6 in culture supernatant were all increased by LPS (100 ng/mL), whereas reduced by 40 ng/mL TSA pretreatment (P < 0.05). TSA pretreatment inhibited LPS-induced up-regulation of TLR4 protein expression. Acetylation of NF-κB/p65(Lys310), which was already increased by LPS, was further enhanced by TSA (P < 0.05). On the contrary, LPS-increased DNA binding activity of NF-κB/p65 was decreased by pretreatment with TSA (P < 0.05). The results suggest that TSA-induced anti-inflammation may be attributed to decreases in the expression of TLR4 and DNA binding activity of NF-κB/p65.
Acetylation ; Animals ; Cell Line ; Hydroxamic Acids ; pharmacology ; Inflammation ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; Macrophages ; drug effects ; metabolism ; Mice ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

OBJECTIVETo investigate the molecular basis for a novel human leukocyte antigen (HLA) allele B*5827.

METHODSDNA from the proband was analyzed by polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) typing. The amplified product was sequenced bidirectionally.

RESULTSAbnormal HLA-B locus was observed and its nucleotide sequence was different from the known HLA-B allele sequences, with highest homology to HLA-B*5820 allele. It differs from HLA-B*5820 by 8 nucleotide substitutions in exon 3, i.e., nt 290 (G > C), nt 346 (T > A), nt 390 (A > C), nt 404 (G > C), nt 413 (C > G), nt 471 (A > G), nt 486 (A > G) and nt 487 (C > A), resulting in an amino acid change from ser > arg at nt 97, phe >tyr at nt 115, ser > arg at nt 130, thr > ala at nt 157 and thr > glu at nt 162. Nucleotide differences of nt 404 (G > C) and nt 413( C > G) did not change amino acid.

CONCLUSIONThe sequences of the novel allele have been submitted to GenBank (access No.GU071234). A novel HLA class I allele B*5827 has been officially assigned by the WHO HLA Nomenclature Committee in Jan. 2010.

Alleles ; Base Sequence ; Cloning, Molecular ; Genotype ; HLA-B Antigens ; chemistry ; genetics ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis, DNA

BACKGROUNDIncreased triglyceride (TG) occurs in patients with acute coronary syndrome (ACS), and apolipoprotein AV (apoAV) has been shown to lower TG levels. In the present study, we investigated plasma apoAV level and its relationship with TG and C-reactive protein (CRP) in ACS patients.

METHODSA total of 459 subjects were recruited and categorized into control group (n = 116), stable angina (SA) group (n = 115), unstable angina group (n = 116) and acute myocardial infarction group (n = 112). Plasma apoAV level was measured by a sandwich ELISA assay.

RESULTSCompared with controls ((100.27 +/- 22.44) ng/ml), plasma apoAV was decreased in SA patients ((76.54 +/- 16.91) ng/ml) but increased in patients with unstable angina ((330.89 +/- 66.48) ng/ml, P < 0.05) or acute myocardial infarction ((368.66 +/- 60.53) ng/ml, P < 0.05). Inverse correlations between apoAV and TG were observed in the control or stable angina groups (r = -0.573 or -0.603, respectively, P < 0.001), whereas positive correlations were observed in the patients with unstable angina or acute myocardial infarction (r = 0.696 or 0.690, respectively, P < 0.001). Furthermore, a positive relationship between apoAV and CRP was observed in the ACS patients but not in the non-ACS subjects.

CONCLUSIONThe plasma apoAV concentration is increased and positively correlates with TG and CRP in ACS patients.

Acute Coronary Syndrome ; blood ; metabolism ; Adult ; Aged ; Apolipoprotein A-V ; Apolipoproteins A ; blood ; C-Reactive Protein ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Triglycerides ; blood

Objective To study and establish the UPLC fingerprints of Lianhua Qingwen Capsules. Methods The samples were separated with a Waters ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) by linear gradient elution. The wavelength for detection was set at 239 nm; mobile phase was set at a flow rate of 0.3 mL/min;the column temperature was set at 30 ℃. Results UPLC fingerprints of Lianhua Qingwen Capsules were established with 32 common peaks. 9 of 32 common peaks were identified, including neochlorogenic acid (peak No.4, source from Lonicerae Japonicae Flos and Houttuyniae Herba), chlorogenic acid (peak No.6, source from Forsythiae Fructus, Lonicerae Japonicae Flos and Houttuyniae Herba), cryptochlorogenic acid (peak No.8, source from Lonicerae Japonicae Flos and Houttuyniae Herba), isoforsythiaside A (peak No.15, source from Forsythiae Fructus), forsythoside A (peak No.20, source from Forsythiae Fructus), quercitrin (peak No.23, source from Houttuyniae Herba), isochlorogenic acid C (peak No.24, source from Lonicerae Japonicae Flos), phillyrin (peak No.26, source from Forsythiae Fructus), glycyrrhizic acid (peak No.31, source from Glycyrrhizae Radix et Rhizoma). The similarities in 10 batches of Lianhua Qingwen Capsules samples were all above 0.96. Conclusion The method is with good precision, repeatability and stability, which can be used as a new means for the quality control of Lianhua Qingwen Capsules.