OBJECTIVE To culture dendritic cells from the peripheral blood ofpatients with laryngeal squamous cell carcinoma. METHODS Peripheral blood was collected from patients under sterile conditions. The mononuclear cells were separated by centrifugation through a density gradient (Ficoll-Hypaque). The adhesion cells, collected from the mononuclear cells, were cultured with interleukin-4 (IL-4, 100ng),granulocyte/macrophage colony stimulating factor (GM-CSF, 100ng) and tumor necrosis factor ? (TNF-?, 100ng) in RPMI 1640 medium for 7 days. The cell morphology was observed under electronic microscope, and the characteristic surface marker CD80 of the cells was analyzed by flow cytometry. MTT colorimetry was employed to assess the proliferation of the mixed lymphocytes. The mixed lymphocytes were stimulated by DC loaded with laryngeal carcinoma antigen. RESULTS A large quantity of cells characteristic of dendritic cell was obtained. Cytometry analysis showed that 33.76 % of mononuclear cells expressed the characteristic marker CD80. After being cultured for 7 days, the expression level of CD80 was significantly increased (69.15 %). Few DC with laryngeal carcinoma antigen could play a role in stimulating lymphocyte proliferation, and the proliferation effect was optimized when the ratio between DC and T cells was 1:5. CONCLUSION A large number of DCs with high purityand sufficient treatment dose can be cultured in vitro from the peripheral blood of patients with laryngeal carcinoma. This technique may play an important role in experimental studies for clinical applications.

Objective To investigate the inhibitory effects and mechanisms of a nature product derivate oridonin on in?vasion of human lung cancer. Methods Human lung cancer A549 and PC-9 cell lines were treated with oridonin. MTS as?say was used to determine cell proliferation. Transwell assay was used to determine the cell invasion, and adhesion assay to determine the cell adhesion. Flow cytometry was used to determine cell cycle. Western blotting and realtime-PCR were used to detect expression levels of CDK1, mTOR, p53, p21, E-cadherin, CD44,β-catenin, uPA, MMP-2/9, p-AKT and p-Src. The luciferase reporter assay was used to detect the NF-κB promoter activity. Results In vitro proliferation, invasion and adhesion of A549 and PC-9 cells were significantly inhibited by oridonin. The cell cycle was halted by G2/M phase, and ex?pressions of E-cadherin, p53 and p21 were promoted, while expressions of CDK1, mTOR, CD44,β-catenin, uPA, MMP-2/9, p-AKT and p-Src and promoter activity of NF-κB were down-regulated. Conclusion Oridonin is able to inhibit the in vitro invasion of human lung cancer A549 and PC-9 cell lines, which might be correlated with its abilities to regulate the ty?rosine kinase activity.

Objective To explore the value of 18 F-fluorodeoxyglucose (i8 F-FDG) PET/CT examination in the differential diagnosis of the gastric cancer and primary gastric lymphoma (PGL).Methods The clinical data of 80 patients with gastric cancer (60 with non-mucinous adenocarcinoma and 20 with mucinous adenocarcinoma) and 47 patients with PGL [22 with mucosa-associated lymphoid tissue (MALT) and 25 with diffuse large B-cell lymphoma (DLBCL)] who were admitted to the Tianjin Medical University Cancer Institute and Hospital from June 2006 to May 2014 were retrospectively analyzed.Spiral CT scan was first done and then followed by PET.The CT value of the lesions,maximum standardized uptake value (SUVmax) of patients and maximal gastrointestinal wall thickness (THKmax) were analyzed by the ANOVA test.The SUVmax comparison between groups was evaluated with the Student-Newman-Keuls.The lesions type was analyzed by the chi-square test.The THKmax and SUVmax among groups were analyzed by the Pearson correlation analysis.Results 18 F-FDG PET/CT imaging of patients with gastric cancer and PGL showed different types of gastric wall thickening,segmental and limited thickening of gastric wall were the main features of gastric cancer and diffuse and segmental thickening of gastric wall were the main features of PGL.The type Ⅰ,Ⅱ and Ⅲ of lesions were detected in 12,21 and 27 of 60 patients with nonmucinous adenocarcinoma,in 2,7 and 11 of 20 patients with mucinous adenocarcinoma,in 8,8 and 6 of 22 patients with MALT and in 13,7 and 5 of 25 patients with DLBCL respectively.There were significant differences in the 4 pathological types of lesions among all the patients (x2 =14.849,P ＜ 0.05).The lymph nodes beneath the renal hilum and at the retroperitoneum were involved in 16 patients with gastric cancer and in 10 patients with PGL,and 7 patients with gastric cancer and 12 patients with PGL were complicated with splenomegalia,respectively,showing a significant difference in the splenomegalia between patients with PGL and gastric cancer (x2=7.506,P ＜0.05).There was no significant difference in the metastasis of lymph nodes beneath the renal hilum and at the retroperitoneum between patients with PGL and gastric cancer (x2=0.178,P ＞0.05).Among 80 patients with gastric cancer,positive 18F-FDG was detected in 79 patients and negative 18F-FDG in 1 patient with T3 stage of mucinous adenocarcinoma.Among 47 patients with PGL,positive 18 F-FDG was detected in 46 patients and negative 18F-FDG in 1 patient with stage Ⅰ of MALT.The CT value of the lesion,SUVmax and THKmax in patients with non-mucinous adenocarcinoma,mucinous adenocarcinoma,MALT and DLBCL were (40 ± 8)HU,(39±11)HU,(41±11)HU,(38±9)HU and 9.9 ±6.6,5.6±1.9,4.6 ±2.9,18.3±7.6 and (2.1 ± 1.2) cm,(1.9 ± 0.9) cm,(1.3 ± 1.1) cm and (2.6 ± 1.5) cm,respectively,showing significant differences in the SUVmax among all the groups (F =26.920,P ＜ 0.05).In the pairwise comparisons,there were no significant difference between the MALT group and mucinous adenocarcinoma group (P ＞ 0.05),and significant differences among the other groups (P ＜ 0.05).The CT value of the lesions and THKmax among all the patients were compared,with no significant differences (F =0.578,4.510,P ＞ 0.05).There were no significant differences in the SUVmax and THKmax among all the patients (r =0.055,0.346,0.226,0.133,P ＞ 0.05).Conclusions There is an important diagnosis value of PET/CT examination in patients with gastric cancer and PGL.The pathological types of the lesions in patients with gastric cancer and PGL are different.The occurrence of splenomegalia in patients with PGL is easier than that with gastric cancer.SUVmax of patients with DLBCL is higher than those with gastric cancer and MALT.FDG uptake in patients with mucinous adenocarcinoma and MALT are not enough,and these may lead to false negative result of PET/CT examination.

Objective:To explore the effects of body mass index (BMI) and gender on primary lung cancer 18F-FDG uptake param-eters, standardized uptake value (SUV), and standard uptake value of lean body mass (SUL). Methods:Data of 50 patients with prima-ry lung cancer confirmed by 18F-FDG positron emission tomography (PET)/computed tomography (CT) were retrospectively analyzed. AW4.6 workstation was employed to measure the SUVmean and SUVmax. Meanwhile, PETVCAR (PET Volume Computed Assisted Reading, GE Healthcare) software was used to automatically measure the SULmean, SULmax, and SULpeak. The SUVmean, SUV-max, SULmean, and SULmax of the liver (central region of the right lobe) were also measured automatically by PETVCAR. Afterward, T/N ratios (lesion SUVmax/liver SUVmean, lesion SULmax/liver SULmean, and lesion SULpeak/liver SULmean) of the lung cancer lesions were calculated. Correlations of the 18F-FDG metabolic parameters with BMI and gender of the patients were analyzed. Results:Liver SUVmean and SUVmax demonstrated significant positive correlations with BMI in all the patients (γ=0.38 and 0.36, P<0.05), and the SUVmean and SUVmax were positively correlated with BMI in male and female groups (γ=0.47 and 0.44, P<0.05), respective-ly. By contrast, no correlation existed between the liver SULmean and SULmax and BMI (P>0.05). No significant correlation was not-ed between the SUVmean, SUVmax, SULmean, SULmax, and SULpeak of the lung cancer lesions and BMI (P>0.05). The correlation trend is the same as that in different gender groups. Only the SUVmax T/N ratio of the lung cancer lesions showed a significant nega-tive correlation with BMI (γ=?0.29, P<0.05). The T/N ratios did not correlate with BMI in the different gender groups (P>0.05). Con-clusion:Patient BMI and gender mainly affect SUV values, particularly SUVmax, by contrast, patient BMI and gender did not signifi-cantly influence SUL and T/N ratio (SUL). Hence, SUL can be more suitable to quantitatively analyze and assess treatment response ob-jectively. This result will be helpful to the clinical application and promotion of PERCIST, which evaluates treatment response mainly by SUL.

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Objective: To evaluate and compare the safety and efficiency of the combined therapy of 89Sr plus zoledronic acid and those of 89Sr-chloride alone, in patients with painful bone metastases. Methods: A total of 87 patients with osseous metastasis were ran-domly divided into treatment groups of 89Sr-chloride alone (group A, 53 patients) and 89Sr plus zoledronic acid (group B, 34 patients). A total of 17 patients in group B received zoledronic acid 2-14 days after 89Sr therapy, and 13 other patients in the group received 89Sr 4-7 days after zoledronic-acid therapy. Pain response and KPS score were evaluated after the different treatments. Results: No obvious bone marrow suppression and liver damage were found in all cases. All patients who received both 89Sr-chloride and 89Sr plus zoledronic acid showed reduced bone pain and total discomfort, as well as improved KPS score, but the response was more pronounced in group B (P=0.047; P=0.036). No statistical differences in pain score and KPS scores were observed between the groups treated with zoledronic acid first and 89Sr therapy first (P=1.000; P=0.667). Comparison of bone pain relief and changes in the KPS score of different primary tumors after treatment with 89Sr-chloride or 89Sr plus zoledronic acid showed no statistical significance. Conclusion: Compared with 89Sr-chloride, treatment with 89Sr plus zoledronic acid was more effective in patients with painful bone metastases. The safety of these two treatments are similar.

Objective To investigate the molecular epidemiology and infectious risk factors of linezolid-resistant Enterococci (LRE) isolates in the First Affiliated Hospital of Chongqing Medical University.Methods Thirteen LRE isolates were collected from 2011 to 2013 and confirmed by broth dilution susceptibility testing.The minimum inhibitory concentrations (MIC) of twelve antimicrobial agents were analyzed using Vitek 2 compact.The molecular epidemiology of LRE isolates was determined by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and the Diversilab.A casecontrol study was conducted for the analysis of risk factors, and Logistic regression was performed to evaluate the independent risk factors.Results All thirteen LRE isolates showed low-level resistance to linezolid, and most of these isolates were resistant to tetracycline, erythromycin and ciprofloxacin.However, they had high sensitivity to penicillin, ampicillin and tigecycline.Sequence type 480 was predominant in the hospital, and three isolates (isolates 3, 4, and 5) from July to September 2013 were found to have the same ST, PFGE pattern and rep-PCR group, indicating the same resistance clone.Admission to intensive care unit (ICU), peripheral vascular disease, males, hypoalbuminaemia, enema and linezolid therapy were identified as significant risk factors for LRE infections.Among these factors, admission to ICU, enema and linezolid therapy were independent risk factors for the acquisition of LRE.Conclusions Thirteen LRE isolates collected in the hospital showed a multidrug-resistant phenotype, and a small-scale prevalence was detected from 2011 to 2013.Therefore, attention should be paid to monitor the LRE in the hospital to decrease the prevalence of LRE infections.

Objective To meet the special needs of the department of radiation oncology, a radiation therapy information management system ( RTIMS) has been developed as a secondary database system to supplement the Varian Varis/Aria since 2007. Methods The RTIMS server was used to run a database and web service of Apache + PHP + MySQL. The RTIMS sever's web service could be visited with Internet Explorer (IE) to input, search, count, and print informations from about 30 workstations and 20 personal computers. As some workstations were installed with Windows and IE in English only, some functions had English version. Results In past five years, as the RTIMS was implemented in the department, some further needs were met and more practical functions were developed. And now the RTIMS almost covered the whole workflow of radiation therapy ( RT) . By September 2011 , recorded patients data in the RTIMS is as follows: 3900 patients, 2600 0utpatient RT records, 6800 progress notes, 1900 RT summaries, 6700 charge records, 83000 workload records, 3900 plan application forms, 1600 ICRT records. etc. Conclusions The RTIMS hased on the workflow of RT has been successfully developed and clinically implemented. And it was demonstrated to be user-friendly and was proven to　significantly improve the efficiency of the department. Since it is an in-house developed system, more functions can be added or modified to further enhance its potentials in research and clinical practice.

Objective To construct a mutant strain of Streptococcus mutans ( S.mutans ) with clpC-deletion and to investigate the role of clpC gene in genetic competence.Methods The fragment of clpC gene and the kanamycin resistant cassette flanked by two loxP sites were amplified by PCR.The purified fragment of clpC gene was cloned into pMD-19T simple vector to construct pCKX1.The pCKX1 vector was digested with ClaⅠ/EcoRⅠ, then blunted and introduced into lox71-KMR-lox66 to obtain pCKX2 vector via homologous recombination.The pCKX2 vector was linearized with SalⅠ and transformed into S.mutans UA159 strain.The positive strains constructed via homologous recombination were screened with kanamycin and transformed with the thermosensitive plasmid pCrePA.The KMR cassette was excised after incubating at 30℃ for 48 hours.Then the pCrePA plasmid was removed after overnight incubating at 37℃for the prepara-tion of clpC-deletion mutant.Total RNA were extracted from the S.mutans UA159 strain and the clpC-dele-tion mutant strain respectively, and then reverse transcribed into first strand cDNA.The target gene frag-ments were amplified by RT-PCR and analyzed by the agarose gel electrophoresis and sequencing.After be-ing verified by PCR and sequencing, the S.mutans UA159 strain and the clpC-deletion mutant strain were re-spectively transformed with E.coli-S.mutans shuttle vector pDL276 to observe the competence development induced by the competence-stimulating peptide (CSP).Results The PCR and sequencing results showed that the pCKX2 vector and the mutant strain with clpC-deletion were constructed successfully via homologous recombination.No clpC gene was detected in the clpC-deletion mutant as indicated by RT-PCR analysis.The formation of competent clpC-deletion mutant was delayed and the competence state was prolonged as com-pared with its parent strains.Conclusion The clpC gene negatively regulated the formation of competent S.mutans.

Objective To evaluate the efficacy and tolerance of preoperative concurrent chemoradiotherapy in the treatment of locally advanced middle-low rectal cancer.Methods From June 2007 to June 2013,51 untreated patients with histopathologically proven rectal cancer (T3/T4 or N (+))were included in this study.Three-dimensional radiotherapy was delivered to the whole pelvic cavity at 45.0-50.4 Gy/25-28 fractions.Two cycles of chemotherapy with FOLFOX4 or XELOX were given concurrently at weeks 1 and 4 of radiotherapy.Surgery was performed at 4-8 weeks after chemoradiotherapy.Adjuvant chemotherapy with FOLFOX4 or XELOX was given within one month after surgery.The Kaplan-Meier method was used to calculate survival rates,and the log-rank test was used for univariate analysis;the Cox regression model was used for multivariate prognostic analysis.Results Fortynine patients completed the preoperative chemoradiotherapy and surgery.The median follow-up was 2.9 years.The overall sphincter preservation rate was 65％;the overall downstaging rate was 59％.Ten (20.4％) of all patients achieved a pathologic complete response (pCR).Grade ≥3 toxicities occurred in 25％ of all patients,and the overall postoperative complication rate was 31％.The 3-and 5-year sample sizes were 24,12,respectively.The 3-and 5-year overall survival rates were 81％ and 69％,respectively;the 3-and 5-year disease-free survival (DFS) rates were 76％ and 60％,respectively;the 3-and 5-year local recurrence-free survival (LRFS) rates were 78％ and 70％,respectively;the distant metastasis-free survival rates were 82％ and 74％,respectively.The multivariate analysis showed that tumor downstaging was an independent prognostic factor for 5-year DFS and LRFS.Conclusions For locally advanced middle-low rectal cancer,preoperative radiotherapy with concurrent FOLFOX4/XELOX chemotherapy can increase pathologic downstaging rate,pCR rate,and sphincter preservation rate.Patients with tumor downstaging may have a better survival advantage.