Multidrug resistance (MDR) plays a major obstacle to successful gastric cancer chemotherapy. The purpose of this study was to investigate the MDR reversal effect and mechanisms of hyperthermia in combination with neferine (Nef) in adriamycin (ADM) resistant human SGC7901/ADM gastric cancer cells. The MDR cells were heated at 42°C and 45°C for 30 min alone or combined with 10 μg/mL Nef. The cytotoxic effect of ADM was evaluated by MTT assay. Cellular plasma membrane lipid fluidity was detected by fluorescence polarization technique. Intracellular accumulation of ADM was monitored with high performance liquid chromatography. Mdr-1 mRNA, P-glycoprotein (P-gp), γH2AX expression and γH2AX foci formation were determined by real-time PCR, Western blot and immunocytochemical staining respectively. It was found that different heating methods induced different cytotoxic effects. Water submerged hyperthermia had the strongest cytotoxicity of ADM and Nef combined with hyperthermia had a synergistic cytotoxicity of ADM in the MDR cells. The water submerged hyperthermia increased the cell membrane fluidity. Both water submerged hyperthermia and Nef increased the intracellular accumulation of ADM. The water submerged hyperthermia and Nef down-regulated the expression of mdr-1 mRNA and P-gp. The water submerged hyperthermia could damage DNA and increase the γH2AX expression of SGC7901/ADM cells. The higher temperature was, the worse effect was. Our results show that combined treatment of hyperthermia with Nef can synergistically reverse MDR in human SGC7901/ADM gastric cancer cells.

OBJECTIVE: To investigate the characters of infection in patients with malignant tumors, especially the distribution, yearly change of pathogens, and pathogen resistance to common antibacterial agents. METHODS: We respectively analyzed the characters of infection in 489 patients with malignant tumors. RESULTS: The respiratory tract was the most frequent infection site (61.1%). The infection was mainly caused by opportunistic pathogens. The Gram-negative bacterias mainly consisted of Pseudomonas aeruginosa, Escherichia coli, and Acinetobacter baumannii (46.3%). The Gram-positive bacteria mainly consisted of Staphylococcus aureus and Staphylococcus epidermidis (29.9%), and the rest 23.8% of the infection was caused by different fungi, mainly consisting of Candida albicans. The ratio of the Gram-negative bacteria resistance to antibiotics such as penicillins, cephalosporins (except ceftazidime), sulfanilamides, tetracyclines and quinolones was higher. The ratio of the Gram-positive bacteria resistance to antibiotics such as penicillins, macrolides and quinolones was higher. The ratio of fungus resistance to antibacterial agents such as fluconazol and itraconazole was higher. The infection caused by fungi obviously increased in the past 5 years. CONCLUSION The infection in patients with malignant tumors is mainly caused by opportunistic pathogens, and the pathogen resistance to antibacterial agents is serious. The infection caused by fungi is increasing.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anti-Bacterial Agents ; therapeutic use ; Communicable Diseases ; complications ; drug therapy ; microbiology ; Drug Resistance, Microbial ; Female ; Humans ; Lung Neoplasms ; complications ; Male ; Middle Aged ; Neoplasms ; complications ; Pneumonia ; complications ; drug therapy ; microbiology ; Pseudomonas Infections ; drug therapy ; Staphylococcal Infections ; drug therapy ; Young Adult

OBJECTIVE: To explore the relation between diabetes mellitus and clinicopathological factors and the incidence of radiation pneumonitis in patients with non-small cell lung cancer. METHODS: The data of 332 patients with non-small cell lung cancer, who were admitted to the Department of Oncology of Third Xiangya Hospital of Central South University between January 2007 and August 2009, were collected retrospectively. The patients were divided into a diabetes mellitus (DM) group (n=45) and a non-diabetes mellitus (NDM) group (n=287). The clinicopathological factors were compared between the two groups. The patients who received radiotherapy were further divided into a diabetes mellitus (DMR) group (n=33) and a non-diabetes mellitus group (NDMR) group(n=287), and the incidence of radiation pneumonitis was compared. RESULTS: A total of 45 patients (13.55%)developed diabetes mellitus. There was significant difference in the body-weight, age and hypertension (P<0.05), while no significant difference in the pathologic factors, such as tumor pathological type, degree of differentiation, and classification of malignant tumors (TNM) stage between the two groups(P>0.05). No significant difference in the irradiation area was found between the DM group and the NDM group(P>0.05). The incidence of radiation pneumonitis in the DMR group was 42.42%(14 out of 33), while 21.31%(39 out of 183) in the NDMR group, with significant difference in the incidence of radiation pneumonitis between the DMR group and the NDMR group(P<0.05). The risk value in the DMR group was 2.721 folds (95%CI, 1.253-5.910) that in the NDMR group in patients with non-small cell lung cancer companied with diabetes mellitus. CONCLUSION Diabetes mellitus is the risk factor of radiation pneumonitis for patients with nonsmall cell lung cancer who receive radiotherapy.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; complications ; pathology ; radiotherapy ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; complications ; radiotherapy ; Female ; Humans ; Lung Neoplasms ; complications ; pathology ; radiotherapy ; Male ; Middle Aged ; Radiation Pneumonitis ; complications ; epidemiology

OBJECTIVE: To analyze the mdr-1 gene promoter methylation and histone acetylation status in both MCF-7/Adr and MCF-7 cells and to preliminarily explore the epigenetic mechanism of multidrug resistance in breast cancer. METHODS: mdr-1 gene promoter methylation status of the 2 cell lines was detected by methylation-sensitive PCR. mRNA expression of DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) was detected by real-time quantitative PCR. Acetylation level of histone H3 and H4 was examined by optical density assay. RESULTS: Promoter hypermethylation of mdr-1 gene was observed in MCF-7 cells whereas hypomethylation was found in MCF-7/Adr cells. Expression of DNMT1, DNMT3a, and DNMT3b mRNA in MCF-7/Adr cells significantly decreased compared with that of MCF-7 cells (P<0.05). H3 and H4 histone acetylation levels of MCF-7/Adr cells were obviously higher than those of the MCF-7 cells (P<0.01). Expression of HDAC1, HDAC2, HDAC7, and Sirtuin type 1 (SIRT1) mRNA in MCF-7/Adr cells was significantly reduced (P<0.05). CONCLUSION Hypomethylation of the promoter region of mdr-1 gene, high H3 and H4 histone acetylation, and low mRNA expression of DNMTs and HDACs may be important epigenetic factors for the development of MDR in MCF-7/ Adr cells.
ATP Binding Cassette Transporter, Subfamily B, Member 1 ; genetics ; Acetylation ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; metabolism ; DNA Methylation ; genetics ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Epigenesis, Genetic ; Female ; Histone Deacetylases ; genetics ; metabolism ; Histones ; chemistry ; Humans ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; metabolism

OBJECTIVE: To determine the effect of different heating Methods combined with neferine(Nef) on the proliferation and expressions of γH2AX and mdr-1/P-gp in MCF-7/Adr breast cancer cells. METHODS: MTT assay was used to determine block heating, water submerged heating, medium heating, and oven heating combined with 10 μg/mL Nef on adriamycin cultured MCF-7/Adr cell proliferation. The mdr-1mRNA expression was detected by real-time quantitative PCR. γH2AX and P-gp expressions were detected by Western blot. RESULTS: The absorbance values of MCF-7/Adr cells in different heating groups at 42 degree and 45 degree were significantly decreased, the mdr-1/P-gp expression was decreased, and γH2AX expression was upregulated compared with those of the 37 degree control group (all P<0.01). The absorbance values (P<0.01) and mdr-1/P-gp expression(P<0.05) were significantly lower and γH2AX expression(P<0.05) was significantly higher in the hyperthermia combined with 10 μg/mL Nef group than those of 10 μg/mL Nef group and hyperthermia group in MCF-7/Adr cells. The water submerged heating group had the lowest P-gp expression and the highest γH2AX expression among different heating groups at 42 degree and 45 degree in MCF-7/Adr cells (P<0.05). CONCLUSION Hyperthermia can increase the cell toxicity of adriamycin to multidrug resistant breast cancer cells. Hyperthermia significantly damages DNA of MCF-7/Adr cells and the higher temperature, the worse effect. Multidrug resistant breast cancer cells may respond differently to the different heating methods. Combined treatment of hyperthermia with Nef can increase the sensitivity in adriamycin chemotherapy.
ATP Binding Cassette Transporter, Subfamily B ; ATP Binding Cassette Transporter, Subfamily B, Member 1 ; genetics ; metabolism ; Antibiotics, Antineoplastic ; pharmacology ; Benzylisoquinolines ; pharmacology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Histones ; genetics ; metabolism ; Hot Temperature ; Humans ; RNA, Messenger ; genetics ; metabolism

Multidrug resistance (MDR) plays a major obstacle to successful gastric cancer chemotherapy.The purpose of this study was to investigate the MDR reversal effect and mechanisms of hyperthermia in combination with neferine (Nef) in adriamycin (ADM) resistant human SGC7901/ADM gastric cancer cells.The MDR cells were heated at 42℃ and 45℃ for 30 min alone or combined with 10 μg/mL Nef.The cytotoxic effect of ADM was evaluated by MTT assay.Cellular plasma membrane lipid fluidity was detected by fluorescence polarization technique.Intracellular accumulation of ADM was monitored with high performance liquid chromatography.Mdr-1 mRNA,P-glycoprotein (P-gp),γH2AX expression and γH2AX foci formation were determined by real-time PCR,Western blot and immunocytochemical staining respectively.It was found that different heating methods induced different cytotoxic effects.Water submerged hyperthermia had the strongest cytotoxicity of ADM and Nef combined with hyperthermia had a synergistic cytotoxicity of ADM in the MDR cells.The water submerged hyperthermia increased the cell membrane fluidity.Both water submerged hyperthermia and Nef increased the intracellular accumulation of ADM.The water submerged hyperthermia and Nef down-regulated the expression of mdr-1 mRNA and P-gp.The water submerged hyperthermia could damage DNA and increase the γH2AX expression of SGC7901/ADM cells.The higher temperature was,the worse effect was.Our results show that combined treatment of hyperthermia with Nef can synergistically reverse MDR in human SGC7901/ADM gastric cancer cells.

Objective:Hepatocellular carcinoma(HCC)prognosis involves multiple clinical factors.Although nomogram models targeting various clinical factors have been reported in early and locally advanced HCC,there are currently few studies on complete and effective prognostic nomogram models for stage Ⅳ HCC patients.This study aims to creat nomograms for cancer-specific survival(CSS)in patients at stage Ⅳ of HCC and developing a web predictive nomogram model to predict patient prognosis and guide individualized treatment. Methods:Clinicopathological information on stage Ⅳ of HCC between January,2010 and December,2015 was collected from the Surveillance,Epidemiology,and End Results(SEER)database.The patients at stage Ⅳ of HCC were categorized into ⅣA(without distant metastases)and ⅣB(with distant metastases)subgroups based on the presence of distant metastasis,and then the patients from both ⅣA and ⅣB subgroups were randomly divided into the training and validation cohorts in a 7:3 ratio.Univariate and multivariate Cox regression analyses were used to analyze the independent risk factors that significantly affected CSS in the training cohort,and constructed nomogram models separately for stage ⅣA and stage ⅣB patients based on relevant independent risk factors.Two nomogram's accuracy and discrimination were evaluated by receiver operator characteristic(ROC)curves and calibration curves.Furthermore,web-based nomogram models were developed specifically for stage ⅣA and stage ⅣB HCC patients by R software.A decision analysis curve(DCA)was used to evaluate the clinical utility of the web-based nomogram models. Results:A total of 3 060 patients were included in this study,of which 883 were in stage ⅣA,and 2 177 were in stage ⅣB.Based on multivariate analysis results,tumor size,alpha-fetoprotein(AFP),T stage,histological grade,surgery,radiotherapy,and chemotherapy were independent prognostic factors for patients with stage ⅣA of HCC;and tumor size,AFP,T stage,N stage,histological grade,lung metastasis,surgery,radiotherapy,and chemotherapy were independent prognostic factors for patients with stage ⅣB HCC.In stage ⅣA patients,the 3-,6-,9-,12-,15-,and 18-month areas under the ROC curves for the training cohort were 0.823,0.800,0.772,0.784,0.784,and 0.786,respectively;and the 3-,6-,9-,12-,15-,and 18-month areas under the ROC curves for the validation cohort were 0.793,0.764,0.739,0.773,0.798,and 0.799,respectively.In stage ⅣB patients,the 3-,6-,9-,and 12-month areas under the ROC curves for the training cohort were 0.756,0.750,0.755,and 0.743,respectively;and the 3-,6-,9-,and 12-month areas under the ROC curves for the validation cohort were 0.744,0.747,0.775,and 0.779,respectively;showing that the nomograms had an excellent predictive ability.The calibration curves showed a good consistency between the predictions and actual observations. Conclusion:Predictive nomogram models for CSS in stage ⅣA and ⅣB HCC patients are developed and validated based on the SEER database,which might be used for clinicians to predict the prognosis,implement individualized treatment,and follow up those patients.

Objective:To investigate the role of the clearance of exogenous myelin antigen in experimental autoimmune encephalomyelitis (EAE).Methods:EAE was induced in C57BL/6J mice by subcutaneous immunization with myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) or FITC-MOG 35-55. The concentration of exogenous myelin antigen was assessed by analyzing the proliferation of the transferred CFSE-labeled mT/mG-2D2 CD4 + T cells in spleen tissues. The release of exogenous myelin antigen from the inoculation sites was analyzed by immunohistochemistry and flow cytometry. HE staining was used to investigate the mechanism underlying the rapid clearance of exogenous myelin antigen. The role of the clearance of exogenous myelin antigen in EAE was investigated by comparative analysis of EAE induced by subcutaneous immunization in the back and footpads, and analyzing the therapeutic effect of soluble MOG 35-55. Results:The proliferation of mT/mG-2D2 CD4 + T cells in mice was enhanced on day 2 than on day 7 after immunization [(52.6±6.8)% vs (18.5±4.9)%, P<0.01]. There was no significant difference in the proliferation of mT/mG-2D2 CD4 + T cells between EAE mice (day 13) and naive mice [(4.4±1.5)% vs (2.5±1.4)%, P=0.11]. Immunohistochemistry and flow cytometry showed that MOG 35-55 was released and engulfed by CD11b + cells at the inoculation sites, and no more MOG 35-55 was released at the onset of EAE. HE staining showed that granuloma that formed surrounding the antigen emulsion during EAE development prevented antigen release from the emulsion, completely isolating the antigen from the peripheral immune system. The incidence of EAE was relatively low in mice immunized via footpads, which was related to the sustained release of MOG 35-55, but had no direct relation to CD4 + regulatory T cells. Continuous intraperitoneal injection of soluble MOG 35-55 could prevent and treat EAE. Conclusions:Exogenous myelin antigen has been completely cleared in EAE mice, and the occurrence of EAE depends on the clearance of the myelin antigen.

Objective To evaluate the prognostic value of anemia in patients diagnosed with extranodal nasal-type natural killer (NK)/T cell lymphoma (NKTCL).Methods Clinical data of 1 225 NKTCL patients receiving the first course of treatment from 10 medical institutions in China were retrospectively analyzed.According to the diagnostic criteria in China,anemia was defined as the hemoglobin (Hb) level＜ 120 g/L for the male and＜ 110 g/L for the female from the sea-level area.The severity of anemia was classified into the extremely severe anemia (Hb ≤ 30 g/L),severe anemia (31-60 g/L),moderate anemia (61-90 g/L) and mild anemia (＞90 g/L).Results Among 1 225 patients,199(16.2％) were complicated with anemia,who had more adverse prognostic factors compared with their counterparts without anemia.Among NKTCL patients with anemia,the proportion of patients with stage Ⅱ-ⅣV,a median age＞ anemia,Eastern Cooperative Oncology Group (ECOG) score of 2-4 and NK/T-cell lymphoma prognostic index (NKTCLPI) ≥ 2 was relatively high.Patients with anemia obtained worse clinical prognosis than those without anemia.The 5-year overall survival (OS) and progression-free survival (PFS) in NKTCL patients with anemia were calculated as 49.4％ and 35.4％,significantly lower compared with 63.3％ and 56.0％ in their counterparts without anemia (both P＜0.01).Single factor analysis demonstrated that anemia,age,ECOG score,group B symptom,lactate dehydrogenase,primary tumor site,primary tumor invasion and staging were the prognostic factors of OS and PFS.Multivariate analysis revealed that anemia was still the independent prognostic factor.Conclusions Anemia is not common in patients with NKTCL and these patients obtain poor clinical prognosis.Anemia is an independent prognostic factor for patients with NKTCL.

Objective To analyze the clinical features and prognosis of extra-nodal nasal NK/T cell lymphoma originated from the larynx. Methods Clinical data of 15 cases of extra-nodal nasal NK/T-cell lymphoma originated from the larynx were retrospectively analyzed. The overall survival ( OS ) and progression-free survival ( PFS) were calculated by Kaplan-Meier survival analysis. The effect of different clinical factors on the clinical prognosis was assessed by univariate analysis. Results Among 15 patients,13 cases were male and 2 female. The median age of onset was 40 years. In 8 cases,the lesions were confined to the larynx,and only 4 cases suffered from cervical lymph node involvement. According to Ann Abor staging, 11 cases were classified as grade I,3 as gradeⅡand 1 as gradeⅢ.The median OS was 28. 0 months and the 5-year OS was 32. 0％.The median PFS was 24. 7 months and the 5-year PFS was 33. 3％.Among 14 patients with stage Ⅰ/Ⅱ,the clinical prognosis of patients receiving combined chemo-radiotherapy was significantly better than those of their counterparts undergoing radiotherapy or chemotherapy alone ( median OS:37. 2 vs. 11. 2 vs.3. 7 months,P=0. 004) . Conclusion Extra-nodal nasal NK/T cell lymphoma originated from the larynx is extremely rare, predominantly in middle-aged male patients. The general condition is relatively favorable. Patients present with multiple lesions in the early stage and relatively poor prognosis. The clinical efficacy of chemotherapy combined with radiotherapy is probably higher compared with that of radiotherapy or chemotherapy alone.