] Objective To explore the role and mechanisms of FGF2 in chemo?resistance in breast cancer. Methods The inhibitors for different signal pathway were used to treat two drug?resistant breast cancer cell lines MCF?7/5?Fu and T47D/5?Fu established in our lab. MTS assay was used to determine chemo?sensitivity and Hoechst stain was used to measure apoptosis. Protein activation and FGF2 protein level in cell culture medium were detected by western blot and ELISA respectively. Results Akt inhibitor MK?2206 (20 nM) and mTOR inhibitor AZD8055 (2 nM) significantly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel, but ERK1/2 inhibitor SCH772984 showed no significant effect. Compared to parent cell lines MCF?7 and T47D, p?Akt and p?S6K (represented as mTORactivity) levels were obviously up?regulated in MCF?7/5?Fu and T47D/5?Fu cell lines, and so do the FGF2 mRNA level and FGF2 protein level from culture medium. Moreover, FGFR inhibitor AZD4547 (4 nM) markedly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel and down?regulated activation of FGFR?Akt?mTOR signal pathway. In agreement, FGF2 protein (10ng/ml) enhanced the chemo?resistance of MCF?7 and T47D cell lines to 5?Fu and paclitaxel and up?regulated activation of FGFR?Akt?mTOR signal pathway. Conclusion Activation of FGF2?FGFR?Akt?mTOR signal pathway promoted chemo?resistance of breast cancer cells.

Objective To investigate the leptin level and its relation with Hp infection in the gastric ulcer(GU).Methods Thirty-one Hp positive GU patients were chosen and divided into two groups,one group included 22 patients with successful eradication therapy on Hp and the other group included 9 patients with unsuccessful eradication therapy;12 Hp negative GU patients underwent anti-ulcer therapy;14 Hp negative normal persons were chosen as control.The BMI,serum leptin level,IL-8 mRNA and protein of gastric issue and symptom score were determined before and three month after the therapy.Results The expression of IL-8 protein and mRNA was obviously higher in Hp positive GU patients before therapy(P

Objective To explore the levels of IL-1β,IL-6 and tumor necrosis factor α (TNF α) in serum and cerebrospinal fluid of patients with spontaneous subarachnoid hemorrhage and their relationship with systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS).Methods The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and TNF α in serum and cerebrospinal fluid of patients with spontaneous subarachnoid hemorrhage were measured with enzyme-linked immunosorbent assay (ELISA).Results The levels of IL-1β,IL-6,and TNFα in serum and cerebrospinal fluid of patients with spontaneous subarachnoid hemorrhage were significantly higher than those of control group (P ＜ 0.05),but the increased time of these cytokines was different.Three cytokines in serum and the cerebrospinal fluid levels of IL-1β and IL-6 but not TNFα were significantly related to SIRS and MODS.Condusions The increased cytokine levels in serum and cerebrospinal fluid of patients with spontaneous subarachnoid hemorrhage may be related to SIRS and MODS,and the measurement of IL-1β,IL-6,and TNFαin serum,and IL-1β and IL-6 in cerebrospinal fluid of patients with spontaneous subarachnoid hemorrhage can be useful to predict and treat SIRS and MODS.

Objective To explore the clinical value and safety of early enteral nutrition support in patients after liver transplantation.Methods We retrospectively analyzed the clinical data of 86 cases who used early enteral nutrition support therapy after liver transplantation between January 2008and October 2011.All of patients were uproot the gastric tube at the first day after the operation,and gradual to the normal diet.The patients who used parenteral nutrition support therapy were as the control group(n＝112).Then we compared the data of patients in the two groups.Results The early enteral nutrition is more useful to the patients after liver transplantation than intravenous nutrition [In the seventh day after the operation,the control group's ALT was (45.2 ± 12.9) U/L,AST was (40.2±9.4) U/L,ALBwas (35.6±2.5) g/L,P＜0.05].The early enteral nutrition also can decrease hospital stay and hospital costs [(14.2±3.4) d,P＜0.05].Conclusion The early enteral nutrition is useful and safe to the patients after liver transplantation.

Objective To investigate the clinical significance of Wilm tumor gene (WT1) expression in breast cancer. Methods Real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method was established for detecting WT1 and GAPDH mRNA expression levels in 110 cases of various breast tumor and the corresponding adjacent normal breast tissue. Normalized WT1 expression level (WT1_N)was determined as a ratio between WT1 and GAPDH for each case. The tumor tissue WT1_N over the normaltissue WT1_N of the same case was calculated as T/N_(WT1) ratio, and T/N_(WT1) value was analyzed with the clinicalpathological parameters. Results The WT1_N expression levels of the 102 breast cancer tissues were significantly higher than those of the adjacent normal breast tissues, with the median WT1_N of 2.38 (ranged from 0.12 to 112.3) and 0.81 (ranged from 0.03 to 11.65) for each (P <0.01), but there were no statistical differences between the WT1_N of 8 benign breast tumors and the nearing normal tissues, with the median WT1_N of 0.46 (ranged from 0.16 to 5.04) and 0.53 (ranged from 0.14 to 4.94) for each. Furthermore the WT1_Nas well as the T/N_(WT1) ratio of the malignant breast cancer tissues were significantly higher than those of the benign tumor tissues, with the median T/N_(WT1) value of 2.54 (ranged from 0.28 to 172.88) and 1.17 (ranged from 0.09 to 2.63) for each. Non-parameter correlation analysis showed that the T/N_(WT1) in breast cancers were of no relevance to lymph node metastasis, clinical-pathological types, estrogen receptor and progestone receptor status, but positively correlated with the expression level of IL-8 gene which calculated with T/N IL-8(r =0.723, P <0.01). Conclusion The WT1 gene is highly expressed in breast cancer which suggests that WT1 level assessed by RQ-RT-PCR could be a novel marker of disease progression and poor prognosis.

This study investigated the intracellular localization of asparagine synthetase (ASNS) in the relation with chemoresistance in leukemia. pIRES-GFP-ASNS-Flag/Neo expression vector was transiently tansfected into SK-N-MC cells and 297T cells respectively. Immunofluorescence and Western blot analysis were performed for cellular localization of ASNS respectively. U937 cells were treated with L-asparaginase for 48 h and examined for endogenous ASNS expression on plasma membrane by immunofluorescence staining. Immunofluorescence staining showed that the transiently expressed ASNS was partly localized on transfected-SK-N-MC cell surface. Moreover, Western blotting exhibited that ASNS expressed both in cytosol and on plasma membrane of transfected-293T cells. Immunofluorescence staining with anti-ASNS-specific monoclonal antibody revealed that endogenous ASNS was localized on the plasma membrane of U937 cells, except for its distribution in the cytosol. In addition, ASNS exhibited a higher expression on plasma membrane after treatment with L-asparaginase as compared with the untreated cells. It was concluded that the subcellular translocation of ASNS may play an important role in L-asparaginase resistance in leukemia cells.

Objective To evaluate the prognostic value of blood lactate (Lac) level in sepsis patients with or without diabetes.Methods 106 patients admitted to intensive care unit (ICU) of Zhongshan Hospital Affiliated to Fudan University from April 2015 to November 2016 were enrolled. The patients with age > 18 years and the length of hospital stay > 24 hours were included. Records including blood Lac, serum creatinine (SCr), white blood cell count (WBC), platelet count (PLT), sequential organ failure assessment (SOFA) on the first day of admission; minimum oxygen index (PaO2/FiO2) in 3 days after admission; mechanical ventilation, whether there was a history of diabetes, usage of biguanides, etiology control treatment, usage of continuous renal replacement therapy (CRRT) were collected. According to the level of blood Lac patients were divided into high Lac group (Lac > 2 mmol/L) and low Lac group (Lac ≤ 2 mmol/L);based on their diabetic history, sepsis patients were divided into the diabetes group and non-diabetes group. The survival curve of each group was analyzed by Kaplan-Meier regression analysis, and the factors influencing the prognosis were analyzed by multivariate Cox regression analysis.Results There were 76 males and 30 females sepsis patients, with an average age of (68.1±14.7) years old. In the 51 patients of low Lac group, there were 7 patients who suffered from diabetes. While in the 55 patients of high Lac group, there were 12 patients who suffered from diabetes. Compared with low Lac group, high Lac group had a higher age, higher SOFA score, and a lower proportion of patients who had the treatment of etiology control (allP < 0.05). There was no significant difference of blood Lac in sepsis patients with diabetes and those without diabetes (mmol/L: 3.03±2.73 vs. 2.81±2.40,P > 0.05). Kaplan-Meier survival curve analysis showed that the 90-day survival rate in the high Lac group was significantly lower than that in the low Lac group (56.36% vs. 90.20%,χ2 = 0.697,P = 0.008). The high Lac group without diabetes had lower survival rate, and the 90-day survival rate was significantly lower than that of the low Lac group without diabetes (58.14% vs. 90.90%,χ2 = 7.152,P = 0.007); there was no significant difference in 90-day survival rate between the high Lac group and the low Lac group with diabetes (50.00% vs. 85.71%,χ2 = 0.012,P = 0.914). Multivariate Cox regression analysis showed that blood Lac was an independent risk factor for the prognosis of sepsis patients [odds ratio (OR) = 3.863, 95% confidence interval (95%CI) = 1.237-12.060,P = 0.020]. After stratification according to their diabetic history, the blood Lac was an independent risk factor for the prognosis of sepsis patients without diabetes (OR = 4.816, 95%CI = 1.407-15.824, P = 0.010), but the blood Lac had no effect on the prognosis of sepsis patients with diabetes (OR = 0.000, 95%CI =0.000-1.103,P = 0.270).Conclusions The predictive value of blood Lac on sepsis patients with or without diabetes was different. The blood Lac was related with the prognosis of sepsis patients without diabetes, while further study should be conducted for the prognostic value of blood Lac in sepsis patients with diabetes, and it's possible to increase the cut-off-point of Lac level in these patients.

Objective To evaluate the clinical efficacy and safety of capecitabine continuous maintenance in treatment of re-currence of triple-negative breast cancer,which had responded to combined chemoradiotherapy.Methods The triple-negative breast cancer was defined as negative of ER,PR and HER2.A total of 46 patients of triple-negative breast cancer were divided into the treatment group (23 patients)and the control group (23 patients).The treatment group was given capecitabine continuous mainte-nance after 6 cycles of chemotherapy.The control group was given 6 cycles of combined chemotherapy only.The clinical efficacy and safety was evaluated between the two groups.Results The PR,PD,RR,and DCR in the treatment group were significantly higher than those in the control group(P <0.05).The acute toxic effects (except for hand-foot syndrome)were similar in two group(P >0.05).Conclusion Capecitabine continuous maintenance after combined chemoradiotherapy in treatment of recurrence of triple-neg-ative breast cancer is more effective,lower toxicity and tolerable.

OBJECTIVETo provide a scientific basis for systematic research on the mechanism of chronic immobilization stress (CIS) induced metabolic network change in rats, through detecting the changes of endogenous metabolites in rats with CIS, treated or un-treated with Xiaoyao Powder (XYP), for determining the small molecule marker compound that closely associated with the metabonomical specificity of CIS and acting mechanism of XYP.

METHODSThirty-six experimental male SD rats were divided into 3 groups, the normal control group, the model group and the XYP group. And all the three groups were subdivided into two subgroups respectively on day 7 and day 21 of the experiment. The stress rat model of CIS was made by chronic restraining method for 3 h every day. Starting from the first day of modeling, XYP 3.854 g/kg in volume of 1 mL/100 g body weight was administered 1 h before restraining via gastrogavage to rats in the XYP group, while equal volume of distilled water was given to rats in the other two groups instead. Blood samples were collected on the 8 th day and 22 th day for detection in the following procedure: at 27 degrees C, 300 microL supernate of blood plasma was taken, calling the Carr-Purcell-Meiboom-Gill (CPMG) and longitudinal eddy-delay (LED) sequence respectively on a Fourier variable nuclear magnetic resonance (NMR) spectrometer, pre-saturated inhibition of the water peak was performed; free induction decay (FID) signals were transferred via 32 k Fourier transformation to gain one-dimensional NMR spectrogram; by taking TSP as the chemical migration reference peak, the segmental integral calculus (0.04 ppm per segment) was performed from 4.5 - 0.5 ppm (CPMG) and 6.0 - 0 ppm (LED) within the peak ranges in 1H spectra using the VNMR software; after normalization, centering and scaling were conducted on data, then used for pattern recognition of principal component analysis (PCA) using the SIMCA-P 10.0 software, or if necessary, the partial least squares discriminate analysis (PLS-DA) was performed.

RESULTS(1) The metabolites in the model group were significantly different from those in the control group, suggesting that the animal model was successfully established with the metabolic network different to that of control. The model group and the XYP group could be differentiated from the control group by the differences of metabolites and metabolic networks between groups; XYP could intervene the metabolites or the metabolic path to cause changes in final metabolites. (2) The serum contents of lactic acid (1.4, 4.16), choline (3.24), N-acetylgalactosamine (NAC) and saturated fatty acids (1-3) increased, but unsaturated fatty acids (1.99,4-5), blood sugar (34), HDL (0.83), etc. reduced in the CIS rats. XYP showed obvious regulatory effects on final metabolites, causing decrease of lactic acid, choline, NAC, saturated fatty acids and blood sugar, and increase of unsaturated fatty acids, blood sugar, HDL, 3.44 ppm compound, etc.

CONCLUSIONSThe metabolic phenotype in CIS rats includes the increase of lactic acid, choline, NAC, saturated fatty acid, and the decrease of blood sugar contents, unsaturated fatty acid, HDL, 3.44 ppm compound, etc., these may be the markers of the metabolites. The final metabolites changes induced by CIS are primarily the lipid substances. XYP markedly regulates the contents of final metabolites, showing the regulatory effects on final metabolites, but what is the metabolite or metabolic pathways it interferes to alter the final metabolites should be confirmed by further studies.

Animals ; Blood Chemical Analysis ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; Powders ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; drug effects

OBJECTIVETo study the antileukemic activity of L-asparaginase through determining the changes of 4 kinds of amino acids (Asn, Aspa, Glu and Gln) in cell culture medium.

METHODSFollowing L-Asp treatment with designed concentrations and duration, the IC50 (inhibitory concentration 50%) of 8 kinds of common leukemia cell lines (U937, HL-60, Jurkat, NB4, THP-1, Namalwa, Karpass299, K562) were determined by CCK-8 assay. The changes of the 4 kinds of amino acids mentioned above were detected by high performance liquid chromatography (HPLC).

RESULTSThe asparagines in cell culture medium were rapidly exhausted when treated with 0.01 U/mL L-Asp for 4 hrs or 1 U/mL L-Asp for 5 minutes. There were significant differences in the sensitivities to L-Asp of different leukemia cell lines. The sensitivities to L-Asp of various cell lines were dose-dependent. Low concentration of L-Asp resulted in a low IC50 and the IC50 increased following the L-Asp concentration increased.

CONCLUSIONSDifferent leukemia cell lines have different sensitivities to L-Asp, suggesting that exhaustion of asparagines around leukemia cells could not reflect the treatment efficacy of L-Asp. L-Asp antileukemic activity is dose-dependent, which suggests the importance of high-dose L-Asp on childhood acute lymphoblastic leukemia.

Asparaginase ; pharmacology ; Asparagine ; analysis ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; Humans ; Leukemia ; drug therapy ; metabolism ; pathology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy