Objective To investigate the significance of optimization of bedside Gram staining of sputum smear in the early diagnosis and antimicrobial treatment for ventilator-associated pneumonia(VAP)patients. Methods The data of patients with VAP undergoing mechanical ventilation over 48 hours in the Department of Critical Care Medicine of Tianjin Fourth Central Hospital from June 2009 to June 2014 were analyzed. The patients were divided into two groups according to whether or not bedside Gram staining of sputum smear was used or not. The sputum samples from lower respiratory tract of all VAP patients were collected daily with tracheal catheter. In empirical examination group(from June 2009 to December 2011,n=43),the patients received antibiotics at the time of onset of VAP, selection of antibiotics depended on the information of bacterial epidemiology of the intensive care unit(ICU),and also existence of high risk factors of multi-drug resistant bacteria. In target treatment group(from January 2012 to June 2014,n=43),the patients received antibiotics according to the results of bedside instant sputum smear examination and empirical antibiotic regime. The correlation between the results of sputum smear examination and culture result was analyzed. The levels of body temperature,white blood cell(WBC)count,procalcitonin(PCT)level,and high sensitivity C-reactive protein(hs-CRP)were measured on the 1st day and 3rd day. The length of antibiotics treatment, duration of mechanical ventilation,and the time of ICU stay were recorded for both groups. Results There were 512 qualified sputum specimens for culture,from which 336 pathogens were found,and 358 strains of pathogenic bacteria were found from microscopic examination of 512 qualified sputum smear. The coincidence rate of results of bedside examination of sputum smear and that of sputum culture was 78.32%(401/512). The diagnostic acumen of the former was 85.42%(287/336),specificity was 64.77%(114/176),positive predictive value was 80.17%(287/358),and negative predictive value was 74.03%(114/154). On the 1st day,no statistical differences in infection index between the two groups could be found,but on the 3rd day,the results were significantly improved in both groups. Compared with the empirical treatment group,the body temperature,WBC,PCT and hs-CRP in the target treatment group were significantly lower〔body temperature(℃):36.83±0.69 vs. 37.64±0.71,WBC(×109/L):7.91±2.75 vs. 9.66±3.39,PCT(μg/L):7.14±3.89 vs. 10.14±4.32,hs-CRP(mg/L):12.24±6.28 vs. 15.54±5.94,P<0.05 or P<0.01〕. Compared with the empirical treatment group,the time of antibiotics use(days:6.00±2.55 vs. 9.20±3.46), the duration of mechanical ventilation(days:5.00±1.73 vs. 7.00±1.94),and the length of ICU stay(days:7.43±1.72 vs. 12.57±4.16)were significantly shortened(P<0.05 or P<0.01). Conclusions The results of bedside sputum examination and sputum culture showed a good correlation,and the former is helpful in early diagnosis and treatment of VAP. The result of high quality sputum smear in significant in guiding the first choice of antibiotics,reduce the time of antibiotic use,shorten the duration of mechanical ventilation and the length of ICU stay,and improve the outcome of the patients.

OBJECTIVETo study the risk factors for capillary leak syndrome (CLS) in neonates.

METHODSThe clinical data of 52 neonates with CLS (case group) were retrospectively reviewed. Fifty hospitalized neonates without CLS were used as the control group. The possible factors for the development of CLS were identified by univariate analysis. The independent risk factors for CLS were determined by multivariate logistic regression analysis.

RESULTSThe univariate analysis showed that the incidences of hyperglycemia, respiratory distress syndrome (RDS), sepsis and cold injury syndrome in the case group were significantly higher than those in the control group (P<0.05). The logistic regression analysis showed that sepsis (OR=5.004, P=0.001), RDS (OR=3.880, P=0.013) and cold injury syndrome (OR=3.207, P=0.023) were the independent risk factors for the development of CLS.

CONCLUSIONSRDS, sepsis and cold injury syndrome are independent risk factors for CLS in neonates. Hyperglycemia may be associated with the development of CLS.

Capillary Leak Syndrome ; etiology ; Female ; Humans ; Infant, Newborn ; Male ; Retrospective Studies ; Risk Factors

OBJECTIVETo evaluate the efficacy and safety of drug-coated balloon (DCB) angioplasty versus uncoated balloon (UCB) angioplasty in treatment of lower extremity arterial occlusive disease (LEAOD).

METHODSRandomized controlled trial comparing DCB and UCB angioplasty for treatment of LEAOD were searched in online databases. Literature screening and quality assessment was carried out according to the established inclusion criteria and exclusion criteria. Restenosis rate at 6 months after surgery, late lumen loss, target lesion revascularization rate, patency rate, mortality rate, and amputation rate at 1 year after operation were compared between DCB group and UCB group using RevMan 5.3 software.

RESULTSEleven trials involving a total of 1853 patients with 2150 lesions were included, with 1110 patients (1288 lesions) in DCB group and 743 patients (862 lesions ) in UCB group. Meta-analysis showed that the restenosis rate at 6 months after the operation (15.2% vs 39.0%; OR: 0.28; 95%CI: 0.17 to 0.48; P<0.00001), late lumen loss (range -0.05 to 0.56 vs 0.54 to 1.7; WMD: -0.57; 95%CI: -0.93 to -0.21), and target lesion revascularization rate at 1 year after operation (13.0% vs 28.1%; OR: 0.39; 95%CI: 0.23 to 0.64; P=0.0002) were significantly lower in DCB group than in UCB group. The patency rate at 1 year after the operation was significantly higher in DCB group than in UCB group (71.8% vs 52.9%; OR: 2.32; 95%CI: 1.21 to 4.43; P=0.001). The mortality rate (4.8% vs 5.0%; OR: 1.00; 95%CI: 0.62 to 1.63; P=0.99) and amputation rate at 1 year after the operation (3.4% vs 2.9%; OR:1.41; 95%CI: 0.74 to 2.70; P=0.30) did not differ significantly between DCB and UCB group.

CONCLUSIONDCB angioplasty is more effective than UCB angioplasty in endovascular treatment of LEAOD with similar treatment safety.

Smoking is the primary cause of lung cancer and is linked to 85% of lung cancer cases. However, how lung cancer develops in patients with smoking history remains unclear. Systems approaches that combine human protein-protein interaction (PPI) networks and gene expression data are superior to traditional methods. We performed these systems to determine the role that smoking plays in lung cancer development and used the support vector machine (SVM) model to predict PPIs. By defining expression variance (EV), we found 520 dynamic proteins (EV>0.4) using data from the Human Protein Reference Database and Gene Expression Omnibus Database, and built 7 dynamic PPI subnetworks of lung cancer in patients with smoking history. We also determined the primary functions of each subnetwork: signal transduction, apoptosis, and cell migration and adhesion for subnetwork A; cell-sustained angiogenesis for subnetwork B; apoptosis for subnetwork C; and, finally, signal transduction and cell replication and proliferation for subnetworks D-G. The probability distribution of the degree of dynamic protein and static protein differed, clearly showing that the dynamic proteins were not the core proteins which widely connected with their neighbor proteins. There were high correlations among the dynamic proteins, suggesting that the dynamic proteins tend to form specific dynamic modules. We also found that the dynamic proteins were only correlated with the expression of selected proteins but not all neighbor proteins when cancer occurred.
Databases, Genetic ; Databases, Protein ; Humans ; Lung Neoplasms ; etiology ; metabolism ; pathology ; Protein Interaction Mapping ; Protein Interaction Maps ; Smoking ; adverse effects ; Support Vector Machine

OBJECTIVETo investigate the role of methylation on E-cadherin inactivation in nasopharyngeal carcinoma (NPC) cell line HNE1 and CNE2, as well as evaluate the inhibitory effect of 5-aza-2'-deoxycytidine (5-Aza-dC) on cell abilities of proliferation and invasion.

METHODSThe expression level of E-cadherin was measured by RT-PCR, Western blot and immunohistochemistry (polymer method), the methyaltion status was analyzed by methylation-specific PCR (MSP), and cell proliferation and invasion were examined by MTT and invasion assay, separately before and after treatment with demethylating agent 5-Aza-dC.

RESULTSThe expression level of E-cadherin was down-regulated compared with the normal tissue, simultaneously partially methylated in gene promoter. Treatment with 20 µmol/L 5-Aza-dC increased the expression of E-cadherin and reduced the methylation degree. Moreover, it also significantly suppressed cell growth (27.6% for HNE1 cells and 34.3% for CNE2 cells, P < 0.05) and invasiveness (37.2% for HNE1 cells and 29.7% for CNE2 cells, P < 0.05).

CONCLUSIONSAberrant methylation around gene promoter region may play an important part in down regulation of E-cadherin in NPC, suggesting a potential therapeutic strategy for demethylating agents such as 5-Aza-dC.

Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cadherins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; DNA Methylation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Promoter Regions, Genetic

Aim To determine the effect of exosomes from lipopolysaccharide-treated human bone marrow mesenchymal stem cells on proportion of Ly6Chigh and Ly6Clow monocytes/macrophages in inflammatory micro- environment. Methods BMSCs were obtained by gra-dient centrifugation, identified and then treated with li-popolysaccharide for 48 h. The exosomes were purified from conditional medium with or without LPS treatment and identified by CD63 protein using Western blot and transmission electron microscope. The diameters and concentration were detected by Nanoparticle Trafficking Analysis ( NTA ) . The monocytes/macrophages were sorted from bone marrow of the mice by magnetic beads. Cells were co-cultured with exosomes for 24 hours, and then treated with LPS for 48 hours. The proportion of Ly6C monocytes/macrophages was detec-ted by flow cytometry. Inflammatory cytokines in cell supernatant were investigated using ELISA. Results BMSCs surface markers CD44, CD90 were positively detected, but CD34, CD45 were not expressed. BM-SCs presented adipogenic differentiation ability. Exo-somes were positively expressing CD63 protein, and NTA showed that the diameters of exosomes were up to (82.4 ± 3.7 ) nm. BMSCs stimulated by LPS pro- duced more exosomes ( P < 0.01 ) . Exosomes from BMSCs with or without LPS treatment could increase the ratio of Ly6Chigh monocytes (P<0.01) and down-regulate the ratio of Ly6Chigh macrophages (P<0.05), and the effect of LPS treated-exosomes was more signif-icant than untreated-exosomes (P<0.05). Moreover, the concentration of IL-6 was also elevated under exo-somes treatment ( P <0.05 ) . Conclusions Human bone marrow mesenchymal stem cells-derived exosomes contribute to the regulation of Ly6Chigh monocytes/mac-rophages, indicating that they could be involved in the therapeutic treatment of inflammatory diseases.

OBJECTIVETo investigate the clinical features of invasive fungal disease (IFD) in children without underlying diseases.

METHODSThe clinical data of 49 children without underlying diseases who were diagnosed with IFD were retrospectively studied.

RESULTSFungal pathogens were detected in 37 (76%) out of 49 patients, including Cryptococcus neoformans (17 children, 46%), Candida albicans (10 children, 27%), Aspergillus (3 children, 8%), and Candida parapsilosis (3 children, 8%). Fungal pneumonia (17 children, 46%) was the most commonly seen disease, with Candida albicans as the major pathogen (9 children, 53%). The 49 children had at least one high-risk factor for infection, including the use of antibiotics, a long length of hospital stay, and invasive procedures. Of all the children, 82% did not respond well to antibiotic treatment or experienced recurrent pyrexia. Among the 24 children who underwent G tests, 17 (71%) showed positive results. All the children were given antifungal therapy, and among these children, 37 (75%)were cured, 3 (6%) were still in the treatment, 5 (10%) died, and 4 (8%) were lost to follow-up.

CONCLUSIONSIn IFD children without underlying diseases, Cryptococcus neoformans and Candida are the main pathogens, and lung infection is the most common disease. Long-term use of high-dose antibiotics may be an important risk factor for fungal infection. The IFD children without underlying diseases are sensitive to antifungal drugs and have a satisfactory prognosis.

Adolescent ; Antifungal Agents ; therapeutic use ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Invasive Fungal Infections ; diagnosis ; drug therapy ; etiology ; Male ; Risk Factors

The cytoplasm of E. coli is a reducing environment where cysteines do not engage in disulfide bonds. Any disulfide bonds that do appear are rapidly reduced through the action of disulfide reducing enzymes such as thioredoxin and glutaredoxin. To study the influence of E. coli cytoplasm on the solubility of recombinant proteins produced in it, bovine fibroblast growth factor (BbFGF), with single disulfide bond, and anti-HBsAg single-chain Fv (HBscFv), with two disulfide bonds, were selected as the pattern molecules of simple protein and complex protein, respectively. pJN98-BbFGF, a BbFGF expressing plasmid based on the vector pET3c, was constructed and transformed into normal host BL21(DE3) and a reductase deficient strain, E. coli Origami(DE3). At the same time, pQE-HBscFv, a HBscFv expressing plasmid was constructed and transformed into M15 [pREP4] and Origami(DE3). The recombinant BbFGF and HBscFv were produced in 2 types of bacteria and their solubilities and bioactivities were determined, respectively. It was found that the majority of BbFGF had formed inclusion body in the cytoplasm of BL21 (DE3) and all of them turned into soluble protein in Origami(DE3). It was also found the productivity of BbFGF in Origami (DE3) was 5% - 10% of the total protein and the value was 15% - 23% in BL21(DE3). BbFGFs produced in 2 recombinant bacteria were purified by cation exchange and heparin affinity chromatography. MTT assay revealed that the bioactivity of BbFGF purified from Origami(DE3) was higher than its counterpart from BL21(DE3). The ED50 of BbFGFs from different bacteria was 1.6ng/mL and 2.2ng/mL, respectively. As far as HBscFvs, both of them formed inclusion body in the cytoplasm of M15 [pQE-HBscFv] and Origami [pQE-HBscFv]. The inclusion body was solubilized in 6mol/L GuHCl, purified with a His-Trap column and then refolded by dialysis step-by-step against buffers containing downtrend concentration of GuHCl. Indirect ELISA was applied to determine the HBsAg binding activity of HBscFvs. It was found there was no obvious difference between the bioactivity of refolded HBscFvs produced from 2 recombinant bacteria. On the other hand, the supernatant of Origami [pQE-HBscFv] lysate displayed weak bioactivity and its counterpart from M15 [pQE-HBscFv] displayed without any bioactivity. The soluble HBsFv in the cytoplasm of Origami [pQE-HBscFv] was purified by cation exchange and immobilized metal affinity chromatography (IMAC) and the yield was 1 - 2mg/L. Those results suggested that modification of the redox environment of E. coli cytoplasm greatly improved the solubility of recombinant disulfide-bonded proteins produced in it. In the next step, we had like to co-express of molecular chaperones or refoldase to raise the yield of soluble recombinant proteins, as well as optimizing the culture condition of the "oxidizing" E. coli.
Animals ; Antibodies ; genetics ; immunology ; metabolism ; Cattle ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; enzymology ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; Fibroblast Growth Factors ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hepatitis B Surface Antigens ; immunology ; Inclusion Bodies ; chemistry ; metabolism ; Oxidoreductases ; genetics ; Plasmids ; genetics ; Protein Engineering ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Solubility

OBJECTIVETo observe the therapeutic effect of shenqi fanghou recipe (SFR) in preventing and treating radiation injury in patients with head and neck tumor.

METHODSOne hundred and forty patients with head and neck tumor, including nasopharyngeal carcinoma, carcinoma of tonsil or tongue, were randomly divided into 2 groups, 70 patients in the observed group were given modified SFR as adjuvant to radiotherapy, while 70 patients in the control group were treated with radiotherapy alone. The radiation reactions during radiotherapy and the condition of late stage radiation injury radiotherapy in patients in the 2 groups were observed.

RESULTSThe degree of oropharyngeal mucosa reaction, dryness in mouth and radiation dermatitis in cervical region in the observed group was milder than those in the control group, and the radiation injury induced late stage sequelae, such as the degree of mouth-opening was better and the cervical muscular sclerosis was better in the observed group than in the control group, showing significant difference (P < 0.01).

CONCLUSIONSFR has definite effect in preventing and treating radiation reaction and late stage radiation injury in patients with head and neck tumor.

Adult ; Aged ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Head and Neck Neoplasms ; drug therapy ; radiotherapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; Radiation Injuries ; drug therapy ; prevention & control