Objective:To investigate the molecular mechanism of transcytosis of Leptospira interrogans ( L. interrogans) across vascular endothelial cells. Methods:Transwell assay was performed to observe the ability of L. interrogans strain Lai across the monolayer of human vascular endothelial cells (HUVEC). Transmission electron microscopy and laser confocal microscopy were used to detect the endocytic vesicles containing L. interrogans strain Lai in HUVEC. The leptospiral endocytic pathway was determined by endocytic inhibition test. Laser confocal microscopy was also used detect the co-localization of L. interrogans with lysosomal marker LAMP1 in HUVEC. The exocytosis of L. interrogans from HUVEC was detected using Petroff-Hausser counting chamber and darkfield microscopy. Results:L. interrogans strain Lai could rapidly transmigrate through HUVEC monolayers and be internalized into HUVEC by PI3K-microfilament-dependent endocytosis to form leptospiral endocytic vesicles. The internalized L. interrogans did not co-localize with LAMP1, indicating the leptospiral endocytic vesicles did not fuse with lysosomes. The exocytosis of internalized L. interrogans was through FAK-microfilament/microtubule pathway. Conclusions:L. interrogans strain Lai could transmigrate through HUVEC by transcytosis to diffuse in vivo and cause disease aggravation.

Objectives To analyze pathogen distribution and antimicrobial resistance of bacilli among children with otitis media. Methods Pathogenic bacteria was isolated from children with suppurative otitis media. The VITEK32 was used for iden-tification. The bacterial susceptibility testing was done by Kirby-Bauer method. According to CLSI standard the antimicrobial susceptibility was determined. Results From Jan 2010 to Dec 2012, 425 children with suppurative otitis media were examined. 347 strains were isolated, of which the detectable rate was 81.65%. The detectable rate of bacteria and fungus was 93.37%(324/347) and 6.63%(23/347), respectively. Among bacteria, the detectable rate of streptococcus pneumoniae was 40.92%(142/347) and staphylococcus aureus was 33.43%(116/347). The detectable rate of haemophilus influenza was 7.78%(27/347). The preva-lence of streptococcus pneumoniae is high in children aged 1-3years, with detectable rate at 47.09%. There was no statistical dif-ference among different age groups. The prevalence of methicillin-resistant staphylococcus aureus (MRSA) in middle ear secre-tion was 1.11%(5/45), 18.75%(9/48)and 30.43%(7/23)in 2010, 2011 and 2012 respectively, with no statistical difference (χ2=3.86, P=0.145). The prevalence of penicillin-resistant streptococcus pneumoniae (PRSP) in middle ear secretion was 9.26%, 3.92%and 27.03%in 2010, 2011 and 2012 respectively, with statistical difference (χ2=11.47, P=0.003). Conclusions Choosing correct therapy according to the result of middle ear secretion culture and antibiotics sensitive test can increase the recovery rate of otitismedia.

Objective To investigate the effect of hyperbaric oxygen preconditioning (HBO) on activation of extracellular signal-regulated kinase (ERK) and death-associated protein kinase 1 (DAPK1)in ischemia-like treated neurons.Methods (1) Cultured mouse primary neurons were allocated into control,ischemia-like condition treated and HBO preconditioning groups.Neurons in ischemia-like condition treated group were treated with 95％ NO2+5％ CO2 for 30 min,and cells in the HBO preconditioning group were pretreated with 98％ O2+2％ CO2 for 2 h,followed by ischemia-like condition treatment.At the end ofthe treatment,the growth of neurons was measured by MTT assay and expression of phosphorylated extracellular signal-regulated kinase (p-ERK) was detected by Western blotting.Neurons were transfected with ERK over-expression vector and pretreated with HBO and ischemia-like condition,and then,the expressions of activated caspase-3 (cleavage caspase-3) and DAPK1 in neurons were analyzed by Western blotting.The interactions of ERK with DAPK1 were also detected by irnmunoprecipitation.(2) Forty-five BALB/c mice were randomized into control,ischemia treated and HBO preconditioning groups.After finishing the treatments,the interactions of ERK with DAPK1 in hippocampal neurons were detected by immunoprecipitation.Results As compared with control group,cells in ischemia-like condition group showed a reduction of cell survivals,while cells in HBO preconditioning group exhibited an inhibition effect on ischemia-like condition-induced cell survival reduction.The expressions of p-ERK were increased by ischemia-like condition,while decreased by HBO preconditioning,with significant difference (P＜0.05).Moreover,over-expression of ERK promoted level of cleavage caspase-3 in neurons; however,DAPK1 expressions were not affected by ischemia-like condition or HBO preconditioning.The interactions of ERK with DAPK1 were attenuated by HBO preconditioning as compared with those by ischemia-like condition treatment.Conclusion HBO preconditioning inhibits ERK activation and disturbs the interaction between ERK and DAPK1,which blocks cell apoptosis and thus induces ischemic tolerance.

Background and objective Afatinib is an irreversible ErbB-family blocker with a clinical activity in non-small cell lung cancer with epidermal growth factor receptor (EGFR) mutations. hTe aim of this study is to assess the safety of afatinib in patients with advanced lung adenocarcinoma. Methods Patients with lung adenocarcinoma (stage IIIb or IV) with EGFR mutations were ifrst-line treated with an oral administration of afatinib (40 mg/d) until disease progression. Adverse events, effects, and survival condition were observed. Results hTe most common adverse events were diarrhea (n=5, 100%), skin rash (n=4, 80%), and mucositis/stomatitis (n=4, 80%). Moderate toxicities not exceeding grade 3 were observed. Relatively, the most serious adverse reaction was mucositis/stomatitis. Mild diarrhea occurred in all patients. hTree patients experienced temporary drug withdrawal and dose reduction because of adverse reaction. Among the four patients who were evaluated, partial response was observed in two patients (50%), one with stable disease (25%) and one with progressive disease (25%). Median progression-free survival was 9.7 months, whereas median overall survival was 18.4 months. Conclusion Afa-tinib was approved as ifrst-line treatment for patients with advanced lung adenocarcinoma. hTe most common adverse events were diarrhea and skin rash. However, mucositis/stomatitis related to afatinib should also be considered. Considering the small number of cases, the conclusion requires more trials for conifrmation.

Objectives To investigate the changes and features of drug resistance in Haemophilus inlfuenzae (Hi) isolated from children with infection diseases in Suzhou. Methods One thousand two hundred and twenty-two Hi strains isolated from clinical specimens were collected from January 2011 to June 2012. Antimicrobial susceptibility was tested by Kirby-Bauer me-thod, andβ-lactamase was analyzed by chromogenic nitroceifn method. Then strains were divided into four groups according to their speciifc resistance patterns:①β-lactamase positive strains (except for cefuroxime-resistant strains),②β-lactamase negative ampicillin-resistant (BLNAR) strains,③cefuroxime-resistant strains, and④other resistant strains. Results The resistance of iso-lated Hi from children in Suzhou area to ampicillin, co-trimoxazole, cefaclor, tetracycline, chloramphenicol, azithromycin, cefu-roxime, ampicillin/sulbactam was 32.7%, 76.7%, 27.4%, 14.3%, 10.2%, 8.4%, 6.9%and 4.3%, respectively, and the detection rate in four groups was 29.0%, 2.6%, 6.9%, 41.2%, respectively. Multi-resistant strains were mainly distributed in Group①and Group②, accounting for 67.5%and 81.3%of each group. Azithromycin, tetracycline and chloramphenicol showed high antimicrobial activity against BLNAR and cefuroxime-resistant Hi strains. The positive rate of resistance toβ-lactamase was 30.8%. The rates of resistance to cefuroxime, cefaclor, ampicillin/sulbactam and cotrimoxazole inβ-lactamase-positive ampicillin-resistant strains are signiifcantly different from those inβ-lactamase-negative ampicillin-resistant strains. Conclusions The resistance feature of Hi isolated from children in Suzhou shows signiifcant changes, including new appearance of cefuroxime-resistant strain, a rapid increase in resistance to azithromycin, and a large proportion of multidrug-resistant strains. The rapid increase in BLNAR and the emergence of cefuroxime-resistant strains have become the new resistance pattern of Hi in this area.

Objective To investigated the efficacy and toxic effects of combined chemotherapy of vinorelbine plus cisplatin or carboplatin in patients aged ≥ 70 years and with non-small cell lung cancer (NSCLC).Methods One hundred patients with lung cancer aged ≥70 years were enrolled in the study.Fifty patients in chemotherapy group were assigned to receive vinorelbine 25 mg/m2 at the first day and the fifth day plus cisplatin 60-70 mg/m2 or carboplatin 250 mg/m2 at the second day.All treatments were repeated every 3 or 4 weeks.Another fifty patients aged ≥ 70 years were taken as control group, not receiving treatment.The primary endpoint was survival.Results Forty-five patients were evaluable for response and the partial remission rate was 35.6% (16/45).One year survival rate was 37.8% and median survival time was 9.75 months in chemotherapy group.The median survival time was 4.0 months for patients in control group.All 50 patients in chemotherapy group were evaluable for toxic side effects.WHO grade Ⅲ incidences of leucopoenia, neutropenia and anemia were 38.0%, 52.0% and 2.2%, respectively.Grade IV incidence of neutropenia was 35.5%.WHO grade Ⅲ incidences of fatigue, constipation and vomit were 22.0%, 8.0% and 14.8%,respectively.Five patients failed to complete the treatment due to side effects.Conclusions Combined chemotherapy of vinorelbine plus platinum drugs is effective and tolerated in patients aged over 70 years with advanced NSCLC.Even patients with stable clinical effects shows benefit of survival time.

BACKGROUNDGefitinib is an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor which is used to treat advanced non-small cell lung cancer, especially adenocarcinoma. The aim of this study is to evaluate the efficacy, side effects and prognostic factors of gefitinib in adenocarcinoma of the lung.

METHODSA total of 26 patients with advanced adenocarcinoma of the lung were enrolled in the study. Gefitinib was orally administered 250mg once daily until disease progression or the occurrence of intolerable toxicity. They were evaluated regularly and their survival was analyzed.

RESULTSIn 26 patients, there was 1 with complete regression (3.8%), 11 with partial response (42.3%), 9 with stable disease (34.6%) and 5 with progression of disease (19.2%). The objective response rate was 46.2% and the disease control rate was 80.8%. The median progression-free survival time was 8.2 months and the median overall survival time was 10.4 months. The 1-year survival rate was 31.6%. Age ( < 70 years old), skin rash and CEA decrease were significantly related to longer survival, however, times of prior chemotherapy and gefitinib treatment stage did not influence the survival. Mean PS (ECOG) was 3.0 before treatment, and 1.8 after treatment. Mean symptom relief time was 5.2 days.

CONCLUSIONSGefitinib is an effective target drug with slight side effect. It can significantly improve quality of life of patients with adenocarcinoma. It can be used as first-line therapy to patients who are not suitable for chemotherapy.

BACKGROUNDMatrix metalloproteinase-9 (MMP-9), endostatin (ES) and vascular endothelial growth factor (VEGF) are important angiogenic regulators for many neoplasms. The aim of this study is to judge clinical and prognostic values of detection of serum MMP-9, ES and VEGF in patients with non-small cell lung cancer (NSCLC).

METHODSSerum levels of MMP-9, ES and VEGF were detected in 92 patients with NSCLC, 50 patients with pulmonary benign disease and 52 healthy controls by ELISA method.

RESULTSThe serum levels of MMP-9, ES and VEGF in NSCLC patients were significantly higher than those in patients with pulmonary benign disease and healthy controls (P=0.000, P=0.000, P=0.000). The sensitivity and specificity of serum MMP-9 was 92.51% and 79.10% with a cutoff value of 117.17 μg/L, 88.32% and 74.25% for ES with a cutoff value of 100.31 μg/L, and 83.40% and 75.63% for VEGF with a cutoff value of 380.32 ng/L. Serum MMP-9 and ES levels were significant prognostic factors for lung cancer patients (P=0.0145, P=0.008). The change of serum MMP-9 level after chemotherapy was a useful indicator of prognosis for NSCLC patients (P=0.0322).

CONCLUSIONSThe serum levels of MMP-9, ES and VEGF are significantly increased in patients with NSCLC. They might be used as prognostic parameters in patients with NSCLC.

BACKGROUNDUroacitides is a group of cell differentiation inducers, which is purified from fresh human urine. Preclinical studies of Uroacitides have showed that cancer cells could be induced to differentiate, and the growth of cancer cells could be inhibited by Uroacitides. The aim of this study is to compare the efficacy and toxicity between Uroacitides combined with NP regimen and NP alone in treatment of advanced non-small cell lung cancer (NSCLC).

METHODSForty-two cases of advanced NSCLC were randomized into Uroacitides+NP and NP groups. NP group: NVB 25mg/m² on days 1 and 8, DDP 75mg/m² on day 1. Uroacitides combined with NP group: Uroacitides of 300mL was given through subclavian catheter daily for 7 days prior to the NP chemotherapy, then concurrently with NP regimen for 2 cycles, except the days of administration of chemotherapy.

RESULTSIn the Uroacitides+NP group, the overall response rate was 44.4%, and 20.0% in the NP group (P > 0.05). The median survival time was 9 months in the Uroacitides+NP group and 6 months in the NP group (P=0.0287). The main toxicities were myelosuppression, gastrointestinal reaction and alopecia, and there was no significant difference in incidences of toxicities between the two groups (P > 0.05).

CONCLUSIONSUroacitides combined with NP regimen shows a good curative effect and low toxicity, and may significantly prolong the median survival time for advanced NSCLC.

BACKGROUNDGensing Rg3 is an active component from ginseng. The aim of this study is to observe the clinical anticancer effect of Rg3 in combination with chemotherapy regimen NP (vinorelbine+cisplatin) in advanced non-small cell lung cancer (NSCLC).

METHODSStage III-IV NSCLC patients confirmed by pathology or cytology all received vinorelbine plus cisplatin for at least two cycles, and were randomized into two groups: patients in arm A also received placebo twice a day, while patients in arm B received two tablets of Rg3 twice a day for at least two months. The endpoints of the study were the efficacy, survival and tolerance of patients.

RESULTSFrom July 2000 to May 2002, 115 patients were enrolled into the trial. The patients' characteristics were well balanced in the two groups. Sex of patients: male, 79; female 36. Types of pathology: adenocarcinoma, 71; squamous cell carcinoma, 29; adenosquamous carcinoma, 8; others, 7. TNM stage: stage III, 45; stage IV, 70. Prior chemotherapy: with, 17; without, 98. Prior radiotherapy: with, 15; without, 100. Prior surgical treatment: with, 23; without, 92. Nine patients discontinued from the trial due to severe adverse effects (5) and other reasons (4), so there were 106 patients evaluable for clinical efficacy. The response rate was 14.5% (8/55) in arm A, and 33.3% (17/51) in arm B (P=0.011). The survival time in arm A was 9.7 months (mean) and 8.0 months (median), and 15.3 months (mean) and 10.0 months (median) in arm B (P=0.0088).

CONCLUSIONSPreliminary results show improvements in response rate and survival time (median and mean) in Rg3 arm compared with placebo arm. It is worthy to confirm the results in further clinical trials.