Objective: To study the mechanism of hypoxia-induced drug-resistance in esophageal squamous cell carcinoma and observe the effect of hypoxia inducible factor 1 alpha (HIF-1α) on the Taxol-induced apoptosis of EC9706 cells. Methods: Small interference RNA (siRNA) targeting HIF-1α was constructed and transfected into human EC9706 cells. Then EC9706 cells were cultured under chemical hypoxia conditions induced by CoCl2, a chemical hypoxia inducer. The expression of HIF-1α in EC9706 cells were detected by Western blot before and after RNA interference. The effect of Taxol on apoptosis of EC9706 cells were determined by TUNEL assay and Annexin V/PI double staining. Results: The expression of HIF-1α protein was induced by CoCl2 75 μmol/L after 4 h, and was significantly inhibited by siRNA-HIF-1α. The apoptosis rate of EC9706 cells induced by hypoxia were significantly higher in siRNA-HIF-1α transfection group than those without transfection or transfected with control siRNA (P < 0.05). Conclusion: Over-expression of HIF-1α inhibited the apoptosis of EC9706 cells induced by Taxol under hypoxia. siRNA targeting HIF-1α increases the therapeutic efficacy of esophageal squamous cell carcinoma.

Objective To evaluate the diagnostic and prognostic value of the serum procalcitonin ( PCT ) level in the non-acquired immune deficiency syndrome ( AIDS ) immunocompromised critically ill patients suspected to have infection. Methods A retrospective study was conducted in the non-AIDS immunocompromised patients who were admitted to Department of Critical Care Medicine of Xiangya Hospital, Central South University during January 2011 to December 2014. Demographic characteristics, underlying disease, acute physiology and chronic health evaluationⅡ( APACHEⅡ) score at admission, and clinical records including baseline and peak levels of temperature, white blood count ( WBC ), PCT, and survival rate within 28 days, infection focus, infectious agents ( bacterial, fungi or mixed infection ), and the severity of infection ( sepsis, severe sepsis, or septic shock ) were recorded. Receiver operating characteristic ( ROC ) curve was plotted, and the diagnostic and protective value of above parameters was evaluated. Results A total of 98 patients ( 43 male and 55 female ) were enrolled in the study with a median age of 44 ( 28, 52 ) years old and a median APACHEⅡscore of 17 ( 11, 20 );47 with malignant hematological tumor, 45 with autoimmune diseases, and 6 post solid organ transplantation. Among them 53 patients ( 54.1%) died within 28 days. Twenty-seven patients were diagnosed as systemic inflammatory response syndrome ( SIRS ) without infection. Among 71 patients with infection, 45 were diagnosed as bacterial infection, 10 with fungal infection, and 16 with mixed infection. Sepsis was diagnosed in 7 patients, severe sepsis in 32 patients , and septic shock in 32 patients .①There was no statistical significance in the baseline and peak levels of PCT and WBC, or baseline level of temperature between the groups of SIRS patients without infection and infected patients. The peak level of temperature was significantly higher in the patients with infection as compared with that of the SIRS without infection patients [℃:39.4 ( 38.9, 40.0 ) vs. 38.8 ( 37.8, 39.2 ), Z=-3.268, P=0.001 ]. It was showed by subgroup analysis that in patients with hematological malignant disease or autoimmune diseases, higher level of body temperature was found in infection group compared with non-infection SIRS group [℃:39.5 ( 39.0, 40.0 ) vs. 39.0 ( 38.4, 39.4 ), Z=-2.349, P=0.019;39.0 ( 38.4, 39.5 ) vs. 38.2 ( 37.0, 38.9 ), Z=-2.221, P=0.026 ].②The baseline level of PCT (μg/L ) were 0.54 ( 0.20, 4.19 ), 2.78 ( 0.50, 9.54 ), 1.00 ( 0.45, 6.89 ), and 0.22 ( 0.07, 1.86 ) in non-infection SIRS patients or the patients with bacterial, fungal, and mixed infection, respectively. The peak level of PCT (μg/L ) were 4.19 ( 1.95, 13.42 ), 12.37 ( 3.82, 45.89 ), 1.82 ( 0.49, 17.86 ), and 5.14 ( 2.66, 12.62 ), respectively, in each subgroup. When the comparison was conducted among the patients with different infectious agent, the baseline level of PCT in patients with bacterial infection was significantly higher than that in SIRS patients without infection ( P=0.026 ) and mixed infection patients ( P=0.001 ), and the peak level of PCT was significantly higher than that in the SIRS patients without infection ( P=0.009 ) and the patients with fungal infection ( P=0.016 ). ROC curve showed that the higher value was found in the baseline and peak levels of PCT for diagnosis of septic shock in all patients [ area under ROC curve ( AUC ) of baseline level = 0.681±0.054, P = 0.001; AUC of peak level = 0.690±0.054, P=0.002 ], and the same value was also found in the baseline and peak levels of PCT for diagnosis of bacterial infection in the patients with malignant hematological tumor ( AUC of baseline level=0.687±0.080, P=0.008;AUC of peak level=0.697±0.079, P=0.021 ).③The peak level of PCT (μg/L ) were 4.05 ( 0.53, 31.22 ), 5.78 ( 2.14, 16.68 ), and 11.64 ( 2.94, 58.14 ) in subgroup of patients with sepsis, severe sepsis and septic shock, respectively, and they showed no statistical significance among subgroups ( P>0.05 ). A high serum level of peak PCT strongly indicated the presence of septic shock ( AUC=0.646±0.060, P=0.019 ), especially in the subgroup of patients with systemic autoimmune disease ( AUC=0.689±0.081, P=0.035 ).④The peak level of PCT (μg/L ) in the APACHEⅡ>18 group ( 38 cases ) was significantly higher than that of APACHEⅡ≤18 group [ 60 cases, PCT (μg/L ):11.64 ( 3.36, 39.39 ) vs. 4.42 ( 1.32, 14.70 ), P=0.016 ];there was a certain correlation between the peak level of PCT and the severity of the disease.⑤The peak level of PCT in death group was significantly higher than that of the survival group [μg/L:9.07 ( 3.05, 33.09 ) vs. 4.19 ( 1.26, 14.61 ), P=0.043 ]. ROC curve showed that the peak level of PCT might be valuable in predicting the prognosis in immunocompromised patients ( AUC=0.619±0.057, P=0.043 ). Conclusions The serum level of PCT is found to be a reliable marker for the diagnosis of bacterial infection in immunocompromised critical patients, especially in those with hematologic malignancy. Additionally, PCT provides a useful tool for evaluating the severity of infection and the prognosis of critically ill patients.

OBJECTIVE To prevent and control nosocomial infection(NI) among patients with hematological malignancies,and make clinical investigation.METHODS The random medical records of patients suffering from hematological malignancies in our hospital were analyzed retrospectively.RESULTS NI rate was 51.11%,in which the main infection site first was respiratory tract,next oral cavity and gastrointestinal tract.Gram-negative bacteria and fungi were the most common pathogens.CONCLUSIONS Patients with hematological malignancies are susceptible to NI,and they should be treated properly so as to prevent from infection effectively.

Objective:To explore the cooradinative training model of postgraduate and and residency of nuclear medicine based on the ability promotion. Develop the learning of basic medical imaging knowledge, the training of clinical practice, the development of scientific research ability and the training of new technology and so on.Methods: Bibliometric analysis was used to analysis the domestic and overseas current situation and research hotspot about carrying out the training of clinical medicine and residency. Combing the existing problem and combined the characteristic of medical imaging, a training model based on ability enhancements be proposed and supporting system is constructed.Results:Search on the Wanfang Database, 170 literatures were obtained from 2007 to 2016 about training of postgraduates and residents of clinical medicine. The annual distribution was increased gradually, the articles number in 2016 was 4.3 times than in 2007. The research hotspots focus on the integration of medical postgraduate student and resident, clinical ability and cultivation of scientific research thinking ability , the penetration of new technology and the choices of research topics. Literature review shows the postgraduates of medical imaging faced the deficiency of anatomy and radiology knowledge, the capacity for scientific research, the training of new radiology technology. Proposed nuclear medicine postgraduate students and residency training cooradinative transformation of the new model and developed a corresponding training curriculum training system.Conclusion: Based on the background of capacity enhancement, the new model of postgraduate and clinicians training mechanism, the operation of collaborative transformation model, reasonable arrangements for the training and scientific research, the focused training, these methods could effectively satisfied both training needs. In order to develop the high-quality talent with both clinical experience and scientific research ability.

Objective To evaluate the impact of comorbidity and age on the tolerance of firstline single-agent chemotherapy in elderly patients with advanced non-small cell lung cancer(NSCLC).Methods Clinical data of 61 elderly patients with advanced NSCLC(aged over 70 years,median age 72 years) receiving first-line single agent chemotherapy were retrospectively analyzed in this study.Performance status(PS) between 0-1 score was in 52 patients,PS 2 score in the other 9 patents.Patients were treated with gemcitabine or docitaxel as the first line chemotherapy,and the median number of chemotherapy cycles was 3.4.Comorbidity was assessed by Charlson comorbidity index (CC1).Patients with CCI equal to 0 were classified as non comorbidity group(n=26),and patients with CCI≥1 were classified as comorbidity group(n=35).Adverse reactions were graded by using the criteria of NCI-CTC v3.0.Results Age and PS could not predict adverse effects of grade 3 or 4.The incidence of hematologic toxicity of grade 3 or 4 was higher in comorbidity group than in noncomorbidity group(40.0％ vs.15.4％,x2 =4.36,P=0.037).The incidences of febrile neutropenia,non hematologic toxicity of grade 3 or 4 and treatment suspension were higher in comorbidity group than in non-comorbidity group.The most common types of comorbidity were diabetes and chronic pulmonary disease.The incidence of non-hematologic toxicity of grade 3 or 4 was increased in patients with chronic pulmonary disease as compared with patients without chronic pulmonary disease(41.4 ％vs.11.5％,x2=6.061,P=0.032).Conclusions The incidences of adverse reactions,especially hematologic toxicity of grade 3 or 4 are significantly increased in patients with comorbidity after singleagent chemotherapy.Evaluation of comorbidity before treatment is helpful to predict the tolerance of single-agent chemotherapy in elderly NSCLC patients.

Flavonoids ; analysis ; Soybeans ; analysis

OBJECTIVE: To explore the possibility mechanism of non-side population cells (NSP) of Hep-2 be induced into stem-like cancer cells by chemotherapy drug--cisplatin. METHOD: Hep-2 cell lines were sorted by fluorescence-actived cell sorting. The acquired NSP cells in trail group were co-cultured with cisplatin for more than 48 hours,while the control group with normal saline(NS). Then identified the percentage of the side population (SP) cells by flow cytometer. The β-catenin, notch-1 mRNA in trial and control group were detected using quantitative realtime PCR, and the β-catenin, notch-1 protein in two groups were compared by Western blot. RESULT: The percentage of side population cells in two groups were (17.16 ± 0.18)%, (10.05 ± 1.20)%, respectively. There was significant difference between two groups (t = 5.844, P < 0.01). The expression of β-catenin, notch-1 was higher in trail group by qRT-PCR; the protein levels of β- catenin, notch-1 was found to inceased in the trail group by Western blot (t = 5.155, P = 0.031; t = 5.977, P = 0.004). Statistical analysis showed significant difference between two groups (P < 0.05). CONCLUSION NSP cells can be differentiated into stem-like cancer cells after being treating with cisplatin. The supposed mechanism is maybe through wnt/β-catenin, notch signaling transduction pathway abnormalities.
Antineoplastic Agents ; pharmacology ; Cell Differentiation ; Cell Line, Tumor ; Cell Separation ; Cisplatin ; pharmacology ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; pathology ; Neoplastic Stem Cells ; drug effects ; RNA, Messenger ; Signal Transduction ; beta Catenin

Animals ; Cell Survival ; Cells, Cultured ; GTP-Binding Proteins ; metabolism ; Isoproterenol ; adverse effects ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, beta-2 ; metabolism

Objective: To evaluate the effect of clusterin with and without leader sequence on overexpression preventing apoptosis in human prostate LNCaP cells. Methods:The plasmid pIRES2 EGFP was used to generate the clusterin expression constructs with full length or without the leader sequence (designated as pIRES2 EGFP/cluac, pIRES2 EGFP/clubc, respectively). Western blot analysis was employed to compare clusterin expression levels in the lysis and supernatant fluid of clusterin transfected LNCaP cells in vitro . The distribution of different functional domains of clusterin in cells was detected with Immunocytochemical staining. The clusterin's protective role of Na 2SeO 3 induced apoptosis in LNCaP cells was examined by flow cytometry (FCM) and fluorescence microscope. Results: Clusterin expression was detected in the lysis and supernatant fluid of pIRES2 EGFP/cluac transfected LNCaP cells, while clusterin was found only in lysis liquid of pIRES2 EGFP/clubc transfected LNCaP cells, but not found in their supernatant fluid. The distribution of cluserin in the plasm of pIRES2 EGFP/cluac transfected cells was aggregative, and on the other hand, clusterin distributed dispersedly in pIRES2 EGFP/clubc transfected cells. Its anti apoptotic property in LNCaP cells was proved by FCM and fluorescence microscope.Conclusion: It is apparent that clusterin plays an important role in preventing apoptosis in prostate cancer, and the presence of the leader sequence is necessary for clusterin's anti apoptotic function.

OBJECTIVE: To investigate the changes of laryngeal cancer Hep-2 cells to cisplatin chemosensitivity after the interference of siRNA of β-catenin gene expression. METHOD: Using a small interference RNA (siRNA) technology interfere β-catenin gene of Hep-2 cells . The mRNA and protein levels of β-catenin in the Hep-2 cells of different groups were detected by qPCR and Western blot. It was divided into siRNA-β-catenin-Hep-2 siRNA group, β-catenin-Neg negative control group and blank control group. Cell proliferation inhibition rate of different concentrations of cisplatin on three groups was detected by MTT assay. Calculate the 50% inhibitory effective concentration IC50 value. Check the change of three groups of cells' apoptosis rate by flow cytometry after the same concentrations of cisplatin stimulation. RESULT: β-catenin-siRNA interference fragment can specifically reduce the expression levels of β-catenin mRNA and protein. qPCR illustrated the expression of mRNA in β-catenin-siR-NA-Hep-2 interference group decreased 70% (P < 0.05) compared with the control group, Western blot results showed that the β-catenin protein expression of interference group (0. 545 ± 0.111) decreased significantly compared with blank control group (1.507 ± 0.139) and negative control group (1.429 ± 0.089), P < 0.05. The IC50 calculation software showed that IC50 of cisplatin on β-catenin-siRNA IC50 interference group is (5.81 ± 0.46)μg/ml, the blank control group is (10.10 ± 1.01) μg/ml, the difference between the two groups has statistical signifi- cance (P < 0.01). Cell apoptosis rate of β-catenin-siRNA interference group was (26.15 ± 0.60)%, significantly higher than the control group (14.16 ± 0.05)%, P < 0.05. CONCLUSION To interfere the expression of β-catenin can effectively enhance the sensitivity of laryngeal cancer cells to chemotherapeutic drugs cisplatin. It provides a theoretical support for the reduction of laryngeal cancer chemotherapy drug cisplatin dosage.
Apoptosis ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Humans ; Laryngeal Neoplasms ; genetics ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; beta Catenin ; genetics