BACKGROUND:Seed cells from different sources have different ability in cell adhesion,proliferation,and differentiation,which can led to bioactive diversity in constructed tissue engineered products. OBJECTIVE:To explore the differentiation ability of different original fetal osteoblasts during constructing tissue engineered periosteum at molecular level. DESIGN,TIME AND SETTING:The contrast observation was performed at the Central Laboratory of Shaanxi Provincial Peoples’ Hospital between July 2007 and July 2008. MATERIALS:The human amnion cells(consent was obtained from the puerperant) were prepared human acellular amniotic membrane(HAAM) . METHODS:Periosteum-origin osteoblasts(POB) and cranium-origin osteoblasts(COB) were seed on HAAM,cultured for 2,4,6,8,and 10 days,and then their total RNA was extracted,which were reversely transcripted to cDNA. The real-time PCR analysis was used to reveal core binding factor ?l(Cbfa1) ,Osterix,and the cycle threshold was also measured. MAIN OUTCOME MEASURES:The expression of Cbfa1,Osterix,as well as osteocalcin. RESULTS:On tissue engineered periosteum,the expression of Cbfa1 in POB was lower than it that in COB(P

OBJECTIVE To investigate the site of nosocomial infection in ICU,distribution and resistance of bacteria in order to make the intervention strategy.METHODS Antimicrobial susceptibility of pathogenic bacteria isolated from nosocomial infection patients in ICU from Jul 2006 to Jul 2008 was performed by Kirby-Bauer method.RESULTS The main pathogens of nosocomial infection in ICU were Gram-negative organisms(48.09%),Gram-positives(38.93%),and fungi(12.98%).The most common pathogens were Staphylococcus aureus,Acinetobacter baumannii,Pseudomonas aeruginosa,fungi,and S.haemolyticus.All strains of S.aureus and S.haemolyticas were antimicrobial sensitive to vancomycin,teicoplanin and linezolid.All A.baumannii strains were antimicrobial sensitive to cefoperazone/sulbactam.They were resistant to other antimicrobial agents.CONCLUSIONS Gram-negative organisms are the main pathogenic bacteria of nosocomial infection in ICU,but the percentage of Gram-positives and fungi is increasing,S.aureus is the most main pathogenic bacterium of nosocomial infection in ICU.S.haemolyticus is also a main pathogenic bacterium.The pathogenic bacteria of nosocomial infection in ICU are highly resistant to the most antimicrobial agents.

Objective To analyze the factors affecting the efficacy of postoperative radiotherapy (PORT) in node-positive non-small cell lung cancer (NSCLC). Methods 480 patients with stage N_1-N_2 NSCLC after radical surgery were retrospectively reviewed. Of them, 267 patients received adjuvant chemotherapy and 121 received PORT. All patients were grouped based on the N stage, tumor size and lymph node positive ratio (the percentage of positive lymph nodes from the detected lymph nodes, LNPR). Group 1 included patients with tumor size ≤3 cm and LNPR ≤33%, group 2 was tumor size > 3 cm or LNPR > 33%, and group 3 was tumor size > 3 cm and LNPR > 33%. The endpoints were the local recurrence free survival (LRFS) and overall survival (OS). Kaplan-Meier method and Cox's proportional hazards regression model were used for the statistic analyses. Results PORT improved the overall survival only in patients with N_2 disease. Both tumor size and LNPR significantly influenced the efficacy of PORT. The 5-year LRFS for patients with vs. without PORT in the group 1, 2 and 3 were 55% vs. 60% (χ~2 = 0.03,P-0.869), 42% vs. 50% (χ~2 =0.31,P=0.547),and 62% vs. 52% (χ~2=4.25,P=0.036), respectively;and the corresponding OS were 22% vs. 50% (χ~2 = 1.65 ,P =0. 199), 26% vs. 22% (χ~2= 0. 13,P=0.786) and 42% vs. 16% (χ~2= 15.33,P=0.000), respectively. Conclusions Tumor size and LNPR significantly impact the efficacy of PORT . For patients with stage N_2 NSCLC , PORT could improve local recurrence free survival and overall survival when tumor size > 3 cm and LNPR >33%.

Objective To investigate the changes of serum apolipoprotein A-I and its clinical significance to acute respiratory infection.Method Totally 44 patients with acute respiratory infection were divided into three groups according to various concentration of Serum apolipoprotein A-I.They were procaleitonin(PCT)＜0.5 ng/ml group,0.5 ng/ml≤PCT＜2 ng/ml group and PCT≥2 ng/ml group.We measured apolipoprotein A-I,C-reactive protein,procalcitonin and albumin within 24 hours after admission.Results With the increase of serumPCT,the production of ApoA-I and albumin were down-regulated,while CRP up-regulated.Conclusions Apolipoprotein A-I has a sound relationship with the acute respiratory infection.It can be used as one of the diagnostic criteria in severe infection patients who have disorders of lipometabolism.

Objective:To comprehensively investigate the prognostic significance of nodal skip metastasis and lobe-specific me-tastasis for patients with IIIa-N2 non-small-cell lung cancer (NSCLC). Methods:A total of 218 completely resected pN2-NSCLC cases with systematic lymph node dissections from 2008 to 2009 at Tianjin Medical University Cancer Hospital were enrolled. Mediastinal lymph node metastasis was subdivided into continuous metastasis and skip metastasis according to whether N1 lymph nodes were in-volved. Mediastinal lymph node metastasis was also classified into extensive metastasis and lobe-specific metastasis on the basis of whether the lymph nodes involved were within or beyond lobe-specific regions. Overall survival (OS) and disease-free survival (DFS) were compared. Results:For the whole cohort, 5-year OS was 21.6%and 5-year DFS was 16.8%. The 5-year OS for patients with skip metastasis or continuous metastasis were 37.6%and 22.0%, respectively (P=0.008). The 5-year DFS of patients with skip metastasis or continuous metastasis were 29.1%and 15.0%, respectively (P=0.022). The 5-year OS of patients with lobe-specific metastasis and ex-tensive metastasis were 38.3%and 20.4%, respectively (P=0.005). The 5-year DFS of patients with lobe-specific metastasis and exten-sive metastasis were 28.4%and 15.1%, respectively (P=0.009). According to the two patterns, patients were subdivided into three sub-groups:Group A (presence of both skip metastasis and lobe-specific metastasis), Group B (presence of skip metastasis only or lobe-spe-cific metastasis only), and Group C (presence of non-skip metastasis and non-lobe-specific metastasis). The 5-year OS of the three sub-groups were 47.1%(Group A), 28.1%(Group B), and 16.6%(Group C) (P=0.001), and the 5-year DFS of these subgroups were 35.2%(Group A), 20.8%(Group B), and 11.2%(Group C), respectively (P=0.002). Multivariate analysis demonstrated that the combined pat-tern was an independent prognostic factor for both OS and DFS. Conclusion:This combined pattern of lymph node metastasis was a strong prognostic factor for IIIa-N2 NSCLC. This pattern should be considered when predicting prognoses and during the selection of patients that will receive postoperative treatments.

Solitary brain metastasis in non-small cell lung cancer (NSCLC) patients was previously considered as Stage IV. Gen-erally, only chemotherapy or radiotherapy rather than surgery was considered the treatment for these patients;hence, achieving the de-sired effect was difficult. In recent years, the treatment benefit for these patients significantly increased. Several patients were even pro-vided the chance of being cured through standardized and individualized treatment by a multiple disciplinary team (MDT). This article introduces two solitary brain metastasis patients with NSCLC who were treated by MDT in Tianjin Medical University Cancer Institute and Hospital. This article aims to explore a complete set of diagnostic and treatment practices for the benefit of more patients.

Objective To discuss the operative strategies for cervical spine fracture combined with ankylosing spondylitis and the deformity of cervicothoracic kyphosis.Methods Retrospectively analyzed 21 cases of cervical spine fracture combined with ankylosing spondylitis and the deformity of cervicothoracic kyphosis,who underwent surgeries from January,2007 to August,2014.Twenty males and 1 female were included.Mean age was (48.6±7.5) years (range,36-65 years).The preoperative American Spine Injury Association (ASIA) classification distribution:2 cases of Grade A,6 cases of Grade B,7 cases of Grade C,5 cases of Grade D,and 1 case of of Grade E.The investigative surgical methods including posterior cervical open reduction,internal fixation and bone graft fusion;posterior cervical open reduction/decompression,internal fixation and bone graft fusion;or combined posterior and anterior cervical decompression,bone graft fusion and instrumentation.The operative time and blood loss were recorded,the clinical therapeutic effect was evaluated by visual analogue scale (VAS) score,ASIA grade improvement,cervical curvature (the angle between a line drawn parallel to the inferior endplate of C2 and a line drawn parallel to the inferior endplate of C7 in the lateral plane of an X-ray image) and radiological assessment (including bone graft fusion condition and internal fixation position).Results One case died from severe pulmonary infection and respiratory failure at the 45th day after operation.All the other 20 patients obtained complete follow-up with a mean time of (39.4±8.76) months (range,25-59 months).7 cases received posterior cervical open reduction,internal fixation and bone graft fusion,average operative time was (92.1±5.4) minutes and average blood loss was (96.1+23.7) ml.9 cases received the posterior cervical open reduction and decompression,internal fixation and bone graft fusion,mean operation time was (121.4± 14.0) minutes and blood loss was (250.0±38.9) ml.3 cases experienced combined posterior and anterior cervical decompression,bone graft fusion and fixation,average operative time was (222.4± 14.9) minutes and average blood loss was (354.3+46.7) ml.Mean of VAS scores at the 3th month postoperatively was 2.76±0.46,significantly lower than the preoperatively VAS scores (8.95±0.36).Mean of VAS scores at the 24th month postoperatively was 1.77±0.39,significantly lower than those at the 3th month postoperatively.The neurological function of spinal cord achieved significant improvement after operations and continued recovered gradually in the subsequent follow up.ASIA Classification distribution at the 24th month postoperatively was:0 case of Grade A,0 case of Grade B,3 cases of Grade C,8 cases of Grade D,9 case of Grade E.Mean of the cervical curvature after operation was-7.1°± 1.9°,significantly higher than preoperatively(-13.4°±3.3°) and did not have any loss of Cobb's angles at 24th month after operation.All the surviving cases achieved solid bone graft fusion.Mean bone graft fusion time was 5.8 months (range,4-9 months).There was no internal fixation loose,fracture and dislocation occurred during follow up.Conclusion All procedures can significantly relieve the painful symptoms,effectively restore the normal cervical alignment and stability,and remarkably improve the neural function of the patients suffering cervical spine fracture combined with ankylosing spondylitis and the deformity of cervicothoracic junction kyphosis.

Objective: To investigate the mechanisms of nuclear export signal of androgen receptor (NESAR) in the regulation of androgen receptor (AR) protein expression and stability in prostate cancer.Methods: The green fluorescent protein fusion protein expression vectors pEGFP-AR(1-918aa), pEGFP-NESAR (743-817aa), pEGFP-NAR (1-665aa) and pEGFP-NAR-NESAR, and lysine mutants of NESAR pEGFP-NESAR K776R, pEGFP-NESAR K807R and pEGFP-NESAR K776R/K807R, were transiently transfec-ted into prostate cancer cell line PC3.Fluorescence microscopy, Western blot and immunoprecipitation were used to detect NESAR regulation of androgen receptor stability.Results: Under the fluorescence microscope, NESAR-containing fusion proteins were cytoplasmic localization, and their fluorescence intensities were much weaker than those without NESAR.The expression levels of NESAR-containing fusion proteins were significantly lower than those without NESAR.The half-lives of GFP-NESAR and GFP-NAR-NESAR were less than 6 h, while the expression of GFP and GFP-NAR was relatively stable and the half-life was more than 24 h in the presence of cycloheximide.The expression levels of GFP-NESAR were significantly increased by proteasome inhibitor MG132 treatment in a dose-dependent manner;in contrast, MG132 did not show any significant effect on the protein levels of GFP.When new protein synthesis was blocked, MG132 could also prevent the degradation of GFP-NESAR in the transfected cells in the presence of cycloheximide, while it had no significant effect on GFP protein stability in the parallel experiment.GFP immunoprecipitation showed that the ubiquitination level of GFP-NESAR fusion protein was significantly higher than that of the GFP control.The mutations of lysine sites K776 and K807 in NESAR significantly reduced the level of ubiquitination, and showed increased protein stability, indicating that they were the key amino acid residues of NESAR ubiquitination.Conclusion: NESAR was unstable and decreased the stability of its fusion proteins.NESAR was the target of polyubiquitination and mediated the degradation of its fusion proteins through the ubiquitin-proteasome pathway in prostate cancer cells.Our research provides a new way to regulate the level and/or activity of AR proteins, thus helping us understand the molecular mechanisms of AR degradation and strict control of AR in the progression to castration-resistance.

Objective: To assess the health risk associated with drinking water in Hangzhou from 2016 to 2017,and to provide evidence for the safety of drinking water . Methods: The monitoring data of 5 genetic toxic substances（arsenic,hexavalent chromium,cadmium,chloroform,tetrachloromethane）and 13 body toxic substances（lead,mercury,selenium,cyanide,fluoride,nitrate,iron,ammonia nitrogen,manganese,copper,zinc,aluminum,volatile phenol）from 36 source water samples,36 finished water samples and 288 tap water samples in the main urban areas of Hangzhou were collected from 2016 to 2017. The health risk of drinking water containing the chemical pollutants mentioned above were assessed based on the evaluation models recommended by United States Environmental Protection Agency . Results: The concentrations of 5 genetic toxic substances and 13 body toxic substances in source water,finished water and tap water were all within the reference limits issued by Standards for Drinking Water Quality（GB 5749—2006）. The carcinogenic risk,non-carcinogenic risk and total health risk caused by the chemical pollutants in the source water were 2.18×10-5/a,7.75×10-9/a and 2.18×10-5/a. The carcinogenic risk,non-carcinogenic risk and the total health risk caused by the chemical pollutants in the finished water were 1.08×10-5/a,3.70×10-9/a and 1.08×10-5/a. The carcinogenic risk,non-carcinogenic risk and total health risk caused by the chemical pollutants in the tap water were 1.96×10-5/a,3.61×10-9/a and 1.96×10-5/a. The carcinogenic risk and total health risk caused by chemical pollutants ranged from high to low in the source water,tap water and finished water. The non-carcinogenic risks ranged from high to low in the source water,finished water and tap water . Conclusion The health risks of 18 chemical pollutants in drinking water in Hangzhou were at a low level,with the greater carcinogenic risk than the non-carcinogenic risk. Hexavalent chromium had the highest carcinogenic risk,while fluoride and aluminum had the highest non-carcinogenic risk.

Obesity is an established risk factor for endometrial cancer. Leptin, a secreted protein of the ob gene by white adipose tissue, plays an important role in the regulation of food intake and energy consumption in the brain and acts as a potential growth stimulator in normal and neoplastic cancer cells. However, a direct role for leptin in endometrial cancer has not been demonstrated. In the present study, the effect of leptin on the proliferation of Ishikawa endometrial cancer cells was investigated as well as the possible mechanism(s) underlying this action in endometrial cancers which express both short and long isoforms of leptin receptors. The expression of leptin receptor (ObRb) in Ishikawa cells was detected by RT-PCR and Western blotting. The cells after serum starvation, were treated by leptin with various concentrations (0, 10, 50, 100, 150 ng/mL) for different durations (6, 12, 24 h). The effect of leptin treatment on cell proliferation was examined by MTT assay. Meanwhile, inhibitory effect of Janus tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) inhibitor AG490 or extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor PD98059 on the proliferation of Ishikawa cells induced by leptin was also studied. Ishikawa cells were treated with 100 ng/mL leptin for various periods (0, 20, 40, 60 min), and the levels of STAT3 phosphorylation and ERK1/2 phosphorylation were examined by Western blotting. The results showed that leptin induced the phosphorylation of STAT3 and the activation of ERK1/2 in a time- and dose-dependent manner in the Ishikawa endometrial cancer cells. Blocking STAT3 phosphorylation with the inhibitor AG490, or blocking ERK1/2 activation by the specific ERK1/2 kinase inhibitor, PD98059, abolished leptin-induced proliferation of Ishikawa cells. In addition, leptin was found to potently induce the invasion of endometrial cancer cells in a Matrigel invasion assay. Leptin-stimulated invasion was effectively blocked by pharmacological inhibitors of STAT3 (AG490) and ERK1/2 kinase (PD98059). These results suggested that leptin promotes endometrial cancer growth and invasiveness by activating STAT3 and ERK1/2 signaling pathways and therefore blocking its action at the receptor level can be a rational therapeutic strategy.