Objective To construct a qseC-deleted mutant strain of E.coli by Red recombination and to study the effect of qseC gene on biofilm formation in the mutants.Methods The chloramphenicolresistant gene flanked by homologues of target genes was amplified by PCR and electro-transformed into E.coli MC1000.When induced by L-arabinose,the plasmid pKD46 could express three recombinant proteins of λ-prophage,which led to the replacement of target gene(qseC) with chloramphenicol-resistant gene.Then the chloramphenicol-resistant gene was eliminated by FLP-promoted recombination events.The biofilm formation of wild-type and mutant strain was detected by crystal violet staining.Results The qseC-deleted mutant of E.coli was confirmed by various PCR and DNA sequencing.Gene qseC was completely deleted.There was no significant difference in growth ability between the qseC mutant strain and the wild-type strain MC1000.The biofilm formation of wild-type and mutant strain was quantified by crystal violet staining.The absorbance determined with a plate reader at 570 nm was 1.00±0.15 and 0.47±0.10 respectively.Conclusion The qseC-deleted mutant of E.coli was constructed successfully.And the qseC gene plays an important role in regulation of biofilm formation in E.coli.

Three hundred and nine community health service (CHS) practitioners were selected with randomized cluster sampling for survey with the job satisfaction questionnaire-occupational stress indicator (OSI).Degree of the job satisfaction was (68 ± 12)%.Position, work experience, income,feeling of recognition, degree of motivation, the scope for career development, the space for personal ambition and the relationship between salary and work load were potential influencing factors of job satisfaction to CHS practitioners.The authors suggest to establish the dynamic mechanisms of satisfaction study and to formulate comprehensive and effective incentive measures for CHS practitioners.

MicroRNAisatypeofsingle-strandednon-codingRNA,directlybondingtothecomplemen-tary target gene mRNA.That leads to the inhibition of mRNA translation of the target molecule in order to reduce expression of the target gene.MicroRNAs are not only involved in the regulation of inflammatory reac-tion,but also play important roles in the formation of neoplasm.The researches showing that the inflammatory reaction interacting with miRNAs results in the differential expressions of microRNAs in lung cancer have been widespreadly concerned.Along with the deepening of the research on the mechanisms of inflammatory reaction and differential expression of microRNAs,it will provide new strategies for the prevention,diagnosis and treat-ment of lung cancer.

Objective To discuss how to practise and consummate to shoot and assess the portfilm for radiotherapy. Methods Double radiography technique was used to take a portfilm. The reference mark points (plumbum points) were added into the field at the first exposure and the cross lines drawn based on the plumbum point images were definited as the reference coordinate to the actual field. Then, the portfilm was compared and analyzed with the simulator planning film. Results The images of both plumbum points and block boundaries were distinct in the portfilm, but the resolution and contrast of the tissue structure image was inferior to the ordinary film from simulator. However, all different structures and positions could be identified in order to assess if the actual field coincided with the radiation planning. Conclusions Although the real-time property of the portfilm is inferior to the electric portal image device (EPID), the double radiography portfilm is feasible as an important quality control method in radiation therapy due to its lower price and easier performance. The design of reference points is the key point in this technique.

Objective To summarize our experience in the prevention and treatment of iatrogenic vascular(trauma).Methods The clinical data of 24 different types of iatrogenic vascular trauma committed from 2003 to 2006 were retrospectively analyzed.Results The 24 cases included 7 cases of superior mesenteric(arteriovenous) trauma,3 cases of portal venous trauma,4 cases of carotid arterial trauma,6 cases of iliac and femoral arterial trauma,and each one of trauma of popliteal artery,axillary artery,renal artery,left gastric artery,respectively.Treated method: Six cases underwent vascular repair,5 cases had vascular anastomosis,2 cases had vascular replacement,3 cases had vascular ligation,2 cases had covered stent implantation under intervention,and other methods included thrombectomy,thrombolysis and packing.Among the 24 cases,22 were cured completely,one patient died from massive hemorrhage 24 hours after operation,and the other died 5 days after operation.Conclusions Iatrogenic vascular trauma can be prevented and its incidence reduced by increased vigilance,clear identification of the anatomy,and accurate and careful operation.Once iatrogenic vascular trauma has occurred,its cause must be determined and then different treatment methods can be(chosen),based on the cicumstaces. When effective treatment technique is not at hand,one should promptly seek outside support or transfer the patient to an advanced hostipal.

Objective To explore the clinical characteristics of and to provide help to the prevention and treatment of malignant chest tumor with nosocomial mixed fungal-bacterial infection.Methods From July 2007 to June 2015,pathogenic bacteria in sputum,blood,urine,chest incision,thoracic and abdominal fluid,and implantable medical biological material were cultivated in 5067 patients with malignant chest tumor suspected with infection.The clinical characteristics,source of specimen and pathogenic bacteria,the types of diseases,medical intervention activities of 142 cases detected with mixed fungal-bacterial infection were retrospectively analyzed.Fesult In 142 patients,104 patients at clinical stage Ⅲ-Ⅳ accounted for 73.2％,and 94 patients used antibiotics more than 14 days (66.2％);104 cases had implanted biological materials (74.7％);96 cases died (67.6％).A total of 167 strains bacteria were isolated.Sixty-one strains of G+ bacteria accounting for 36.5％ were mainly Epidermis staphylococcus and Staphylococcus aureus;106 strains of G-bacteria accounting for 63.5％ were mainly klebsiella pneumonia,Escherichia coli and baumanii;172 strains fungus mainly of Candida albicans were isolated (77.3％).Pathogenic bacteria sources were mainly sputum specimens + pharynx strip,blood culture and medical implant materials.In 72 lung cancer patients,squamous carcinoma and small cell carcinoma were 52.8％ and 33.3％ respectively,higher than adenocarcinoma (12.5％);In 42 esophageal cancer patients,postoperative patients were 42.9％.Parenteral nutrition patients with more than 10 days were 80.9％ higher than that of parenteral nutrition in patients with less than 10 days (19.1％).Conclusion Among malignant chest tumor patients with nosocomial mixed fungal-bacterial infection,the bacteria were found in staphylococcus aureus,klebsiella pneumoniae and E.coli and the fungus was Candida albicans.For clinical stage Ⅲ-Ⅳ,patients with parenteral nutrition for more than 10 days,having history of chemo or radiotherapy,with antimicrobial use for more than 14 days,and with implanted biological materials,should be warned about nosocomial mixed fungal-bacterial infection.

Objective To investigate the serum levels change of vascular endothelial growth factor (VEGF) during operation of non-small-cell lung cancer (NSCLC). Methods 120 cases of NSCLC patient diagnosed by pathology as well as with operation indication were selected as the experimental group. The patients selected were required without any chemotherapy or radiotherapy before operation, besides that, they should have good compliance and free will to be examined. During the process of experiment, 60 cases concluded as healthy in the physical examination were chosen as control group. The correlative information of the experimental group were collected including periphery blood specimen collected in 3 days before the operation, and of the 1st day, 7th day and 30th day after the operation while the periphery blood specimen of control group were collected. The serum levels of VEGF were detected by adopting enzyme linked immunosorbent assay (ELISA) method. Results The serum levels of VEGF in NSCLC patients before operation, of the first postoperative day, of the seventh postoperative day and of the thirtieth postoperative day were significantly higher than that in healthy people (P＜0.01), respectively (279.14±44.89)μg/L, (282.70±42.74) μg/L, (353.79±44.55) μg/L, (178.40±43.43) μg/L and (91.40±16.55) μg/L. The serum levels of VEGF in NSCLC patients showed positive correlation with the stages (P＜0.01). Conclusion The serum levels of VEGF in NSCLC patients scale up. At the same time it shows positive correlation with the stages of the primary tumor. The serum levels of VEGF in NSCLC patients scales up by degrees in one week after the operation, and drop one month later.

Objective To study the effect of epinephrine on biofilm formation of the qseC-deleted mutant of Escherichia coli on biomaterial.Methods The strains used in this study are Escherichia coli MC1000 and MC1000AqseC.LB was used for all the experiments.To determine the effect of epinephrine on motility,halos were measured in LB medium at 37℃ in the presence of epinephrine(50 μmol/L).LB with epinephrine and without epinephrine were used,and then the experiment of bacterial biofilm formation on PVC material was taken.The relative amount of biofilm was estimated.The thickness of bacterial community and bacterial community quantity in the unit area on PVC materials were measured by confocal laser scanning microscope( CLSM),and the surface structure of biofilm formation was observed by scanning electron microscope(SEM).Results The mutant strain formed less biofilm than the wild-type strain in LB.The increment in motility of wild-type strain due to epinephrine addition was shown,but mutant strain is unaffected.Similarly,biofilm formation of the wild-type strain was increased by epinephrine,but epinephrine did not affect the biofilm formation of the qseC mutant.The CLSM and SEM showed that epinephrine stimulated biofilm formation of wild-type strain on PVC materials,but had no effect on qseC-deleted mutant strain.Conclusion Epinephrine increases Escherichia coli biofilms on biomaterials through qseC.

Objective To investigate the role of quorum sensing Escherichia coli regulator C (qseC) in intestinal bacterial translocation in rats subjected to hemorrhagic shock.Methods Thirty Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 5 groups (n =6 each):control group (group C),MC1000-sham shock group (group M-SS),MC1000qseC-sham shock group (group △-SS),MC1000-hemorrhagic shock group (group M-HS),and MC1000△ qseC-hemorrhagic shock group (group △-HS).The rats drank 150 μg/ml of disinfect water containing streptomycin in 3 consecutive days to inhibit the autochthonous flora in the intestinal tract.From 4th day,the rats were fed with Escherichia Coli MC1000 or MC1000△ qseC 1 ml/100 g by gastric perfusion once a day for another 3 consecutive days in the other 4 groups,while the rats were fed with normal saline instead in group C.Hemorrhagic shock was induced by blood-letting.The mesenteric lymph node (MLN),spleen and liver specimens were obtained at 24 h after operation for bacterial culture and the bacteria were identified.Bacterial translocation from gut to MLN,spleen and liver was observed and the number of bacteria in MLN,spleen and liver tissues were counted.Results The rate of bacterial translocation was significantly higher,and the number of bacterial colonies in MLN,spleen and liver tissues and the total number of bacterial colonies were significantly larger in groups M-HS and △-HS than in group C,and in group M-HS than in groups M-SS and △-SS (P ＜ 0.05).The rate of bacterial translocation was significantly lower,and the number of bacterial colonies in MLN,spleen and liver tissues and the total number of bacterial colonies were significantly smaller in group △-HS than in group MHS.Conclusion QseC is involved in the intestinal bacterial translocation following hemorrhagic shock in rats.

BACKGROUND: Permanent or transient implantation of biomaterials can result in biomaterials-centered infections (BCI) in lung cancer patients.OBJECTIVE: To investigate the relationship between BCI and peripheral blood transforming growth factor β1 (TGF-β1) in patients with lung cancer.METHODS: A total of 248 lung cancer patients undergoing in vivo intravascular catheter indwelling > 7 days were included.Quantitative method was used for intubation, bacteriological culture and paired blood culture, and API Staph strips were adopted for positive patients. While enzyme-linked immunosorbent assay was used to detect TGF-β1 levels in the peripheral blood of patients with lung cancer and 75 healthy volunteers as normal controls.RESULTS AND CONCLUSION: Among the 248 patients, there were 82 BCI-positive cases, and 166 BCI-negative cases.Thirteen patients were confirmed to have catheter-related bloodstream infection. There were 48 Gram-positive bacteria, 24Gram-negative bacilli, and 10 fungal. The levels of TGF-β1 were higher in BCI-positive patients than BCI-negative patients (P < 0.05); the levels of TGF-β1 in the BCI-negative group were higher than those in the normal control group (P < 0.05). For lung cancer patients with nosocomial infection induced BCI, there are various species of pathogenic bacteria, and Gram-positive bacteria are more common. To detect TGF-β1 levels in patients with lung cancer is of significance for early prevention of BCI.