Objective To observe the effect and mechanism of imiquimod on T helper (Th) cell subsets in the Parabronchial lymph node (PBLN) cell cultures from ovalbumin (OVA)-sensitized rats.Methods PBLN were isolated and cultured. PBLN cells were divided into A~F, according to different concentrations of intervention. Cultured for 0, 3, 6, 12, 24, 48 hours, the expressions of IL-4 and IFN-? in supernatants were determined by ELISA. The mRNA expressions of the cytokines in cells were detected by RT-PCR.Results In the group A, only low concentrations of IFN-? were detected. Based on the cultured time, the concentrations of IFN-? were increased significantly if imiquimod concentration was between 1 and 10 ?g/ml. Levels of IL-4 were increased slowly compared with those in the group B (P0.05).Conclusion Imiquimod show the best effect on antigen-specific Th cell subsets when cultured for 12h. The results suggest that imiquimod have benefit in atopic diseases such as the late inflammation reaction of asthma.

Objective To explore drug resistance and distribution of multidrug-resistant(MDR)Mycobacterium tuberculosis (M.tuberculosis)in a county-level hospital,so as to strengthen the prevention and control of health-care-associated infection with M.tuberculosis .Methods Specimens with positive sputum smear were performed M. tuberculosis culture and drug resistance testing,and distribution of MDR tuberculosis patients in the departments before isolation were investigated retrospectively.Results Of 488 patients with tuberculosis,254 were positive for sputum smear,122 M.tuberculosis strains were isolated from positive sputum smear patients,120 isolates were per-formed drug susceptibility testing,results revealed that 86 isolates were drug-resistant strains,46 of which were monodrug-resistant,40 were MDR.Of MDR strains,16 were all resistant to isoniazide,rifampicin,streptomycin, and ethambutol.The percentage of monodrug-resistance,MDR,pandrug resistance was 9.43%,8.20%,and 3.28% respectively.Medical imaging department,ultrasound department,and respiratory disease department were the main units of M.tuberculosis exposure.Conclusion The percentage of MDR M.tuberculosis is high among M. tuberculosis ,surveillance should be intensified,so as to prevent the transmission in hospital.

Objective To investigate the drug resistance and risk factors of hospital-acquired pneumonia (HAP) induced by imipenem-resistant Acinetobacter baumannii.Methods Clinical data on 114 patients with Acinetobacter baumannii-related HAPs admitted in Wujiang First People' s Hospital in Suzhou during January 2013 and December 2014 were retrospectively analyzed.According to the results of drug sensitivity test,patients were divided into imipenem-resistant group and non imipenem-resistant group.Drug resistance to 20 commonly used antibiotics was observed in two groups,and multivariate Logistic regression analysis was performed to identify the risk factors of imipenem-resistant Acinetobacter baumannii infection.Results Among 114 strains ofAcinetobacter baumannii,66 strains (57.89％) were imipenem-resistant and 48 strains (42.11％) were non-imipenem-resistant.The resistance rates to β-lactams,quinolones and aminoglycosides were significantly higher in imipenem-resistant group than those in non-imipenem-resistant group (P ＜ 0.01),and no tigecycline-resistant strain was found in both groups.Univariate analysis showed that acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score ≥ 15,plasma level of albumin ≤ 25 g/L,intensive care unit (ICU) stay,indwelling gastric tube,deep venous catheterization,establishment of artificial airway,mechanical ventilation time ≥ 7 d,use of broad-spectrum antibiotics ≥ 14 d and combined use of antibiotics were risk factors of imipenem-resistant Acinetobacter baumannii related HAP (x2 =13.06,6.86,25.40,15.09,17.87,21.46,17.94,6.91 and 10.10,P ＜0.01).Multivariate Logistic regression analysis revealed that establishment of artificial airway [OR =72.014,95％ confidetial interval (CI):19.566-265.061,P ＜ 0.01],and use of broad-spectrum antibiotics ≥ 14 d (OR =3.892,95％ CI:1.092-13.879,P ＜ 0.05) were independent risk factors of imipenem-resistant Acinetobacter baumannii related HAP.Conclusion Imipenem-resistant Acinetobacter baumannii strains are highly resistant to most antibiotics.Strict control of invasive procedures and long-term combined use of antibiotics may reduce the occurrence of imipenem-resistant Acinetobacter baumannii related HAPs.

Objective To investigate the effects of 4.1N expression in lung cancer A549 cell line on cell proliferation, invasion and migration. Methods A549 cells were cultured in vitro and transfected with lipofectamine 2000 mediation. Three groups were employed: transfection with pEGFP-4.1N plasmid, pEGFP vector plasmid, and blank control, respectively. The mRNA and protein expression differences of 4.1N was examined by semi-quantitative RT-PCR and Western blot in every group after 48 h. The proliferation capability was determined by MTT assay. Invasion capability was evaluated by scratches, adhesion experiments and Transwell chamber model. Results After the transfection, the expression of 4.1N mRNA and protein in pEGFP-4.1N plasmid transfection group was significantly enhanced (P<0.05). The proliferation capability of A549 cells descended extremely (P<0.05). The migration and invasion capability of A549 cells in vitro decreased substantially (P<0.05). Conclusions Transfected with 4.1N gene can significantly increases the expression levels of 4.1N mRNA and protein in A549 cells which are highly metastatic in human. Cell behavior in vitro studies showed that 4.1N gene can inhibit the proliferation, adhesion, invasion and migration of A549 cells, which plays an important role in the metastasis of lung cancer and it may become a molecular marker for metastasis of lung cancer.

Objective To investigate the relationship between risk factors and recurrent respiratory tract infections( RRTI) in children. Methods A case-control study was carried out in 204 children with RRTI,com-pared with 400 children without RRTI in control group. Results Second-hand smoking,breastfeeding,malnutri-tion,zinc deficiency,pet at home,home decoration and use of glucocorticoid were associated with RRTI in chil-dren. But the association between breastfeeding and RRTI was only found in children less than 5 years old. The second hand smoking was associated with RRTI only among boys,rather than girls. Home decoration,zinc defi-ciency,malnutrition and breastfeeding were associated with RRTI only among girls,but not among boys. Conclu-sion Recurrent respiratory tract infections are associated with multiple factors. Different risk factors have differ-ent affections on RRTI on gender.

Objective To investigate the influencing factors of pain and the changes of vital signs in newborn infants. Methods Forty two newborn infants were rated by the behavioral scale of acute pain in newborn infants. The scores of pain were compared among infants with different gender,gestational age,birth weight,birth age,type of puncture and whether by vaginal birth or not. At the same time,the respiration rate,heart rate,blood pressure and oxygen saturation (SO2)were dynami-cally recorded by the multi-function monitor in the process of puncture. Results The average score of pain was 7.6. There was no significant difference among newborn infants with different gender,gestational age,birth weight and type of puncture(P> 0.05),while significant differences among infants with different birth age and whether by vaginal birth or not (P=0.015 and 0.043 respectively). In the process of puncture,the SO2 was significantly decreased,while the respiration rate,heart rate,sys-tolic and diastolic blood pressure were significantly increased. Conclusions Pain is prevalent in newborn infants and accom-panied by obvious changes of vital signs. The means of childbirth and birth age have significant influence on the neonatal pain. It is suggested to pay close attention to the neonatal pain and take effective interventions.

Objective To investigate whether maternal antibody to hepatitis B surface antigen (anti-HBs)in infants may interfere with the antibody response to hepatitis B vaccine. Methods Infants from singleton pregnant mothers,who delivered at full term at the Affiliated Drum Tower Hospital of Nanjing University Medical School from October 2006 to January 2007,were divided into two groups based on their mothers'status of anti-HBs(43 positive and 29 negative).All infants were vaccinated with hepatitis B vaccine at birth and one month thereafter.Serum anti-HBs were quantitatively determined for the mothers before delivery and for infants in cord blood at delivery and in serum at the age of 1 and 3.5 months. Results Anti-HBs of all 43 newborns in the positive group were positive in cord blood with the coefficiency of 0.98 to the maternal serum anti-HBs level(t=39.05,P<0.01).Forty-two out of the 43 infants remained anti-HBs positive at the age of 1 month.Anti-HBs was negative both at birth and 1 month old in infants of the negative group.However,all infants in both groups were anti-HBs positive at 3.5 months of age,while the average concentration of anti-HBs in infants of the negative group was significantly higher than that of the positive group [(466.9±86.7)mIU/ml vs(151.2±23.1)mIU/ml,t=2.72,P=0.011].Among the 5 infants whose maternal anti-HBs level>1000 mIU/ml,3 did not produce active antibodies against two doses of hepatitis B vaccination. Conclusions Passively acquired maternal anti-HBs in infants can inhibit the active antibody response to hepatitis B vaccine,and the extent of this effect is associated with maternal anti-HBs level.

Objective To establish a method for the determination of quetiapine fumarate in human serum by RP‐HPLC and apply it into clinical .Methods Extracting with ethyl ether after serum‐drug was alkalized ,and then determined by RP‐HPLC .The determination was performed on Zorbax Eclipse XDB‐C18 column with mobile phase consisted of methanol‐water (70∶30 ,containing 0 .5% triethylamine and 0 .4% glacial acetic acid) at the flow rate of 0 .6 ml/min .The detection wave‐length was set at 254 nm ,and the column temperature was 35 ℃ .The method would be applied into analysis of clinical medica‐tion .Results Quetiapine fumarate and the impurities could be completely separated ,and the linear range of quetiapine fumar‐ate were 50‐1 000 ng/ml(r=0 .999 5) .The recovery of the method was 98 .2%‐100 .1% and the recovery of extracting was 75 .2%‐84 .6% .RSD of intra‐day was within 0 .8%‐3 .7% and RSD of inter‐day was within 1 .4%‐5 .1% .The limit of quantita‐tion for quetiapine fumarate was 2 .1 ng/ml .This method had been applied into clinical pharmacy and achieved a good effects . Conclusions The method is simple ,accurate ,reproducible ,and sensitive for determination of quetiapine fumarate in human se‐rum .It has important significance on instructing the rational use of clinical medicine and discovering the unreasonable drug combination .

In view of the characteristics of the computerized system,the key points in the quality assurance (QA) of the computerized system was discussed and summarized combined with the requirements of the GLP laboratory in Europe and America.The validation of computerized system,the control during the use of computerized system,period maintenance and safety protection of computerized system,archives of electronic data was discussed,expecting to provide reference for the management of computerized system in Chinese GLP laboratory which is generally not high currently.The experiences were obtained as follow:Through repeated inspection and review,the problem was found and set as the risk point;a targeted QA inspection plan was made focusing on the risk-based inspection and the QA inspection plan was timely adjusted according to the problems,which ensures the pertinence and validity of the QA inspection.

Objective To evaluate the application of fractional exhaled nitric oxide (FeNO) in the reducing treatment of bronchial asthma.Methods From October 2015 to September 2016,60 asthmatic patients with FeNO＞25 ppb were randomized into FeNO group and control group with 30 cases in each group.Patients in both groups were treated with combined inhaled corticosteroids and long-acting beta 2 agonist (ICS/LABA) starting with low doses;the dosage was adjusted according to the symptom control alone in control group,while in FeNO group the dosage was adjusted according to the symptom control and FeNO level.After 1 year-follow up,the Asthma Control Test (ACT) scores,Asthma Life Questionnaire (mini AQLQ)scores,pulmonary function,FeNO levels,blood eosinophil counts,total IgE,hierarchical control level,cumulative corticosteroid use and cumulative months of leukotriene receptor antagonists (LTRA) use were compared before and after treatment within group,and between two groups.Stratified analysis was carried out in the patients complicated with allergic rhinitis.Results After treatment,ACT scores,mini AQLQ scores and FEV1/pred (％) were significantly higher than those before treatmentin both groups (t=10.755,10.189,8.632 and 13.311,8.102,12.456,respectively,all P＜0.05),while the FeNO,EOS and total IgE levels were significantly lower than those before treatment (t=8.005,3.313,3.924 and 8.967,3.885,3.270,respectively,all P＜0.05),and the numbers of patients with good control were significantly increased (Z=-5.035 and-4.976 respectively,P＜0.05).Compared with control group,FeNO level was lower,mini AQLQ scores of symptom scores and emotional scores were higher and the average numbers of asthma attacks per patient per year were less after treatment in FeNO group (t=2.912,4.214,4.589,U=2.154,all P＜0.05).However,there was no significant difference in cumulative corticosteroid use and cumulative months of LTRA use between two groups (U=564.000 t=1.921 and 0.165,respectively,P＞0.05).For patients complicated with allergic rhinitis,the numbers of acute asthma attack were increased and the cumulative dosage of systemic corticosteroid use was higher in control group than those in FeNO group (both P＜0.05).Conclusion The reducing treatment strategy based on FeNO level and symptom control is of clinical value for patients with bronchial asthma,especially for those complicated with allergic rhinitis.