Objective To evaluate the prophylactic efficacy and safety of posaconazole against invasive fungal disease(IFD)in hematologic patients with neutropenia.Methods Medical records of 18 hematologic patients with neutropenia received posaconazole for preventing IFD in a Beijing hospital between 2014 and 2015,the efficacy and safety was evaluated.Results There was no clinical diagnosis or confirmed diagnosis of IFD among 18 patients during posaconazole prophylaxis period,none of patients stopped posaconazole due to severe adverse reaction.Two patients with acute myeloid leukemia(AML) died of pulmonary infection,1 of whom isolated Stenotrophomonas maltophilia from sputum culture on the 12th day of posaconazole prophylaxis,the other isolated Escherichia coli from sputum culture on the 14th day of posaconazole prophylaxis.Other patients all adherence to posaconazole prophylaxis until granulocyte count recovered,patients were followed up until 100 days medication,no death occurred.The lowest peripheral neutrophil count was(0.00-0.27) x 109/L during posaconazole prophylaxis period,with the median of 0.02 x 109/L;the duration of posaconazole prophylaxis was 8-27days,with the median of 16 days;among patients without IFD breakthrough or received systemic use of other antifungal agents,there were 2 (11.1％) all-cause death within 100 days;there were no adverse reaction,such as liver function abnormalities≥grade 2 and kidney function abnormalities≥grade 2,as well as QTc prolongation.Conclusion Posaconazole is effective for preventing IFD in hematologic patients with neutropenia,adverse reaction is rare.

Objective To investigate the effect and mechanism of CRM197, the heparin binding-epidermal growth factor-like growth factor (HB-EGF) inhibitor, on the reverse of the resistance ofovarian cancer to paclitaxel. Methods (1)The effect of CRM197 on the 50% inhibitory concentrations (IC50) of human ovarian carcinoma cell line A2780 and paclitaxel-resistant ovarian carcinoma cell line A2780/Taxol was tested by methyl thiazolyl tetrazolium (MTT) assay. Western blot was used to detect the effect of CRM197 on the expression of HB-EGF,epidermal growth factor receptor (EGFR) and plasma membrane glycoprotein (P-gp) protein in A2780 and A2780/Taxol cells. Real-time PCR was used to examine the MDR1 mRNA expression in these cells. (2) A2780/Taxol cells were divided into 4 groups, including the cells transfected with empty vector and saline treatment (empty vector group), MDR1 small interference RNA (siRNA)vector and saline treatment (MDR1 siRNA group),empty vector and CRM197 treatment (empty vector+CRM197 group) and MDR1 siRNA vector and CRM197 treatment (MDR1 siRNA+CRM197 group), respectively. Flow cytometry was used to detecte the effect of intracellular rhodomine 123 (Rh123) accumulation, and caspase-3 activity assay was used to test the effect of apoptosis in four groups of A2780/Taxol cells. (3) In experiments in vivo, A2780/Taxol cells were inoculated to nude mouse subcutaneously to determine the EGFR and P-gp protein expression following CRM197 treatment by immunohistochemistry. Results (1)In vitro,MTT examination showed that the IC50 of A2780/Taxol cells to paclitaxel in A2780/Taxol+CRM197 group [(6.4±0.3)μmol/L] was significantly lower than the IC50 in A2780/Taxol group [(34.1± 0.5)μmol/L,P<0.01], and the reveral fold of CRM197 was 5.3. The expression level of HB-EGF protein in A2780/Taxol+CRM197 group (1.44 ± 0.29) was significantly lower than HB-EGF protein in A2780/Taxol group (2.72 ± 0.32),respectively (P<0.05). The expression level of EGFR protein (0.71 ± 0.25) and P-gp protein (0.82±0.19) in A2780/Taxol+CRM197 group was significantly lower than EGFR protein (1.87±0.31) and P-gp protein (1.84 ± 0.27) of A2780/Taxol group (P<0.05). Compared with A2780/Taxol group(1.78 ± 0.27), MDR1 mRNA was significantly down-regulated in A2780/Taxol+CRM197 group (0.79 ± 0.13,P<0.05). (2) The fluorescence intensity of Rh123 of the A2780/Taxol cells in empty vector group, MDR1 siRNA group,empty vector+CRM197 group, MDR1 siRNA+CRM197 group was 33.4±1.6, 56.3±3.3, 43.5± 3.1,100.4 ± 7.4, and the pNA of the A2780/Taxol cells was(11.4 ± 1.2),(52.8 ± 0.9),(71.2 ± 3.6),(82.7 ± 3.8)μmol/L. The expression levels in MDR1 siRNA+CRM197 group were both higher than the expression levels in empty vector+CRM197 group, and the expression levels in empty vector+CRM197 group, MDR1 siRNA group were both higher than the expression levels in empty vector group (P<0.05). (3) In vivo, the expression scores of EGFR protein in A2780/Taxol+CRM197 tumors (4.4 ± 1.4) were lower than that in A2780/Taxol tumors (10.2 ± 3.1,P<0.05). The expression scores of P-gp protein in A2780/Taxol+CRM197 tumors (3.8 ± 1.1) were lower than that in A2780/Taxol tumors (8.8 ± 2.7,P<0.05). Conclusion CRM197 reverses the resistance of ovarian cancer to paclitaxel by increasing caspase-3 activity to advance apoptosis via EGFR/MDR1/P-gp pathway.

Objective:To study influence of renal sympathetic denervation (RDN)on cardiac function of dogs with heart failure (HF).Methods:A total of 40 dogs were randomly and equally divided into RDN group [received bilat- eral renal artery radiofrequency ablation (RFA)]and model group (only received femoral puncture).Pacemaker was implanted in every dog,and dog HF model was established using rapid right ventricular pacing.Cardiac and re-nal function indexes,BNP and sympathetic activity index levels were observed and compared between two groups be- fore RFA/sham operation,instant and four weeks after model establishment.Results:After operation four weeks, compared with model group,there were significant reductions in levels of epinephrine (E)[(362.69±42.54)ng/ml vs.(290.36±42.32)ng/ml],renin (R)[(305.46± 39.68)ng/ml vs.(230.04±32.80)ng/ml],aldosterone (AD)[(408.00±38.56)ng/ml vs.(246.00± 48.37)ng/ml],angiotensin Ⅱ (ATⅡ)[(280.00±48.08)pg/ml vs.(172.00±25.04)pg/ml]and norepinephrine (NE)[(425.65±50.54)ng/ml vs.(316.76±46.29)ng/ml]in RDN group (P<0.05 all);there were significant reductions in HR,respiratory rate (RR)and BNP level in RDN group,P<0.05 all;there were significant rise in SBP,LVEF,CO,CI,left ventricular pressure maximal rising rate (+dp/dtmax),left ventricular pressure maximal dropping rate (-dp/dtmax)and left ventricular end-systolic pressure (LVESP),and significant reductions in left ventricular end-systolic dimension (LVESd),left ventricular end-diastolic dimension (LVEDd)and left ventricular end-diastolic pressure (LVEDP)in RDN group,P<0.05 all.Conclusion:RDN can decrease renal sympathetic activity,improve heart function,inhibit myocardial remode- ling,its therapeutic effect is significant

Objective To examine the expression of heparin binding-epidermal growth factor-like growth factor (HB-EGF) in paclitaxel-resistant ovarian cancer and elucidate the relationship between HB-EGF and the resistance of ovarian cancer to paclitaxel. Methods The human ovarian carcinoma cell line A2780 and the paclitaxel-resistant human ovarian carcinoma cell line A2780/Taxol were cultured in vitro. Western blot was used to dectect the expression of HB-EGF protein in A2780 and A2780/Taxol groups. The A2780 cells were treated with cross-reacting material 197 (CRM197 and A2780 + CRM197 group) or dimethyl sulphoxide (DMSO;A2780 group), while the A2780/Taxol cells were treated with CRM197 (A2780/Taxol+CRM197 group) or DMSO (A2780/Taxol group). The effects of CRM197 on growth and proliferation was tested by methyl thiazolyl tetrazolium( MTT) and the results were showed as absorbance (A).The effects of CRM197 on cell cycles was tested by flow cytometry, while the effects of CRM197 on apoptosis was examined by caspase-3 activity assay and the results were showed as p-nitroaniline(pNa). In animal experiment, four groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability following CRM197 treatment. Immunohistochemistry was used to determine the expression of HB-EGF protein in A2780 and A2780/Taxol group. Results The expression level of HB-EGF protein in A2780/Taxol group (2.11±0.41) was significantly higher than that of A2780 group (0.75±0.20;P<0.01). The inhibition effect of CRM197 on the cell growth of A2780+CRM197 and A2780/Taxol+CRM197 group was accompanied by the acceleration of CRM197 concentration(P<0.01). When CRM197≥1 μg/ml, the inhibition effect of CRM197 on the cell growth of A2780/Taxol+CRM197 group was significantly higher than that in A2780/Taxol group(P<0.05). In cell cycle experiment, CRM197 induced the cell-cycle arrest at the G0/G1 phase in A2780+CRM197 cells[(67 ± 4)%] compared with A2780 cells[(54 ± 6)%;P<0.01], while CRM197 significantly induced the cell-cycle arrest at the G0/G1 phase in A2780/Taxol+CRM197 cells [(72± 4)%] compared with A2780/Taxol cells [(24±8)%;P<0.01]. CRM197 treatment in A2780+CRM197 group [(40 ± 6)μmol/L] led to the acceleration of the caspase-3 activity when compared to A2780 group [(6 ± 6)μmol/L;P<0.01], while CRM197 treatment in A2780/Taxol+CRM197 group [(66 ± 12)μmol/L] led to significant acceleration of the caspase-3 activity when compared to A2780 group [(9 ± 6)μmol/L;P<0.01]. In experiments in vivo, the expression scores of HB-EGF protein in A2780/Taxol tumors(10.8 ± 3.3) were higher than that in A2780 tumors (5.0±2.2;P<0.01). The tumor size and tumor weight of the A2780/Taxol+CRM197 group were both higher than those of the A2780+CRM197 group [(546 ± 85) mm3 vs (1 355 ± 119) mm3,(0.56 ± 0.09) g vs (1.31 ± 0.27) g; all P<0.01]. The CRM197 inhibition rate of the A2780+CRM197 and A2780/Taxol + CRM197 group were 43% and 68% respectively, showed that CRM197 significantly suppressed the growth of A2780/Taxol xenografts in vivo(P<0.01). Conclusions HB-EGF is over-expressed in paclitaxel-resistant ovarian cancer and may be contributes to drug resistance. Inhibition of HB-EGF expression potently enhances apoptosis and inhibit the growth of paclitaxel-resistant ovarian cancer, shedding light on the HB-EGF-targeted therapy options for chemoresistant ovarian cancer patients.

Objective: To observe influence of long-term exercise training on carotid intima-media thickness (IMT) in patients with mild-moderate hypertension. Methods: A total of 92 patients with essential mild-moderate hypertension were randomly divided into routine treatment group (n=46) and exercise group (n=46, received exercise training based on routine treatment) according to number table. All patients were followed up for one year, and the blood pressure, carotid artery diameter and carotid IMT were measured, compared and analyzed in two groups before and after treatment. Results: There was no significant difference in systolic and diastolic blood pressure before treatment between two groups (P＞0.05). Compared with baseline value, there were significant decrease in systolic blood pressure [(176.66±l1.78)mmHg vs. (130.89±13.01)mmHg], diastolic blood pressure [(101.43±6.41)mmHg vs. (81.71±8.45)mmHg], carotid artery diameter [(6.62±0.97)mm vs. (6.22±1.01)mm] and carotid IMT [(0.98±0.12)mm vs. (0.84±0.11)mm] in exercise group after one-year training (P<0.05 all), and they were all significantly lower than those of routine treatment group (P<0.05 all ). Conclusion: Long-term exercise training can effectively control blood pressure, decrease carotid artery diameter and carotid intima-media thickness.

Objective PSGL-1 is specifically expressed in leucocytes.The aim of this study was to explore the changes of myeloid-derived suppressor cells (MDSCs) in the spleen and bone marrow in PSGL-1-deficient mice.Methods PSGL-1 -/-mice were used in the experiment.After identification of the offsprings, flow cytometry was used to test the expression of CD11b and Gr-1 in C57 and PSGL-1 -/-mice.Results Compared with the C57 mice, the expression of MDSCs was up-regulated in the PSGL-1-deficient mice ( P <0.001).Conclusion The expression of MDSCs is upregulated in PSGl-1-deficient mice.

Objective To explore the significance in judging the different clinical stages of relapsing polychondritis (RP) patients through examining the changes of aggrecanase and metabolic fragments of aggrecan.MethodsIn comparison with the control group (20 cases),40 patients were divided into the stable stage group (22 cases) and the active stage group (18 cases).The aggrecanase-generated neoeptitopes in cartilage matrix were analysed by immunohistochemistry and Western blot(WB) respectively.The mRNA and protein levels of aggrecanase-1,2 expressed in cartilage cells were measured by real-time reverse transcriptional polymerase chain reaction(RT-PCR) and WB respectively.The difference of these results among these three groups was analyzed accordingly.ResultsThe expression of aggrecanase-1,2 in mRNA level was measured by real-time RT-PCR.The values of aggrecanase-1,2 mRNA 2 -ΔΔC1 were 1.00 ± 0.26 and 1.00 ± 0.27 in control group,1.47 ± 0.11 and 1.57 ± 0.13 in stable stage group,2.09 ±0.12 and 2.09 ± 0.19 in active stage group respectively.By one-way ANOVA analysis,the difference between every two groups was statistically significant (F was 299.113 and 459.013,P ＜ 0.01 ).In comparison with control group,aggrecanase-1,2 increased significantly in both stable and active stage group (P ＜ 0.01 ) and aggrecanase-1,2 increased more significantly in active stage group than in stable stage group (P ＜ 0.01 ).The results from WB analysis indicated that aggrecanase-1,2 could not be detected in control group,and they were detectable in stable stage group and increased in active stage group at the relative molecular of 68 000 Da or 73 000 Da respectively.The aggrecanase-generated neoeptitopes were analyzed by WB as well.The results indicated that NITEGE and ARGSV could be detected in stable stage group and increased in active stage group at the relative molecular of 70 000 Da or 48 000 Da respectively,but there were no signals in control group.Similar with the previous WB results,no signals of NITEGE or ARGSV eptitopes were detected in normal cartilage matrix ( no red staining) by use of immunohistochemical staining.However,in stable stage group and active stage group,these eptitopes were apparently detected (obviously red staining).ConclusionWith the progression of the RP,the activity of the aggrecanase is enhanced,and the degradation of the aggrecan is increased,associated with the severity of the disease.

Objective To investigate the effect of unfractionated heparin (UFH) on the expression of heme oxygenase-1 (HO-1) in intestinal mucosa of mice with sepsis.Methods Thirty-six male C57BL/6J mice were randomly divided into sham group,cecal ligation and puncture (CLP) group and UHF group,n =12 in each group.Model of intestinal injury in sepsis was induced by CLP.In sham group,the mice were exposed without ligation of cecum.In UFH group,the mice were treated intravenously with 8 U of UFH via the tail vein half an hour before the operation and 12 hours after the surgery respectively.Six mice in each group were randomly chosen at 4 hours and 24 hours after operation to collect inferior vena venous blood samples and terminalileum tissues.The serum levels of interleukins (IL-1 β,IL-6),and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA).The serum level of D-lactate was determined by colorimetry.Pathological changes of ileum tissue and Chiu score were observed after hematoxylin eosin (HE) staining.The HO-1 expression was detected immunohistochemically.Results In sham group,no significant changes in the serum levels of IL-1 β,IL-6,TNF-α and D-lactate were observed.Twenty-four hours after the operation,the structure of intestinal mucosa was basically normal without obvious pathology change and no HO-1 positive cells were found.The serum levels of IL-1 β,IL-6,TNF-α,and D-lactate in CLP group were gradually increased,and they were significantly increased as compared with sham group [IL-1 β (ng/L):40.87±2.88 vs.22.60±2.05 at 4 hours,113.73±3.96 vs.22.07±2.74 at 24 hours;IL-6 (ng/L):63.89±3.26 vs.44.89±3.38 at 4 hours,176.56±5.45 vs.45.76±4.02 at 24 hours;TNF-α (ng/L):194.62± 14.13 vs.152.05±6.22 at 4 hours,599.62± 10.20 vs.155.90± 14.18 at 24 hours;D-lactate (mmol/L):0.24± 0.02 vs.0.19 ± 0.01 at 4 hours,0.33 ± 0.04 vs.0.20 ± 0.02 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,edema and inflammation in ileal mucosa,intestinal villi structural damage were observed,the Chiu score was significantly higher than those in the sham group [4.5 (3.0-5.0) vs.0 (0-1.0),P ＜ 0.05],and a small amount of HO-1 positive cells were localized in the intestinal mucosa.Compared with CLP group,the serum levels of IL-1 β,IL-6,TNF-α,and D-lactate of UFH group were significantly decreased [IL-1 β (ng/L):31.53 ± 2.90 vs.40.87 ± 2.88 at 4 hours,61.13 ± 2.80 vs.113.73 ± 3.96 at 24 hours;IL-6 (ng/L):51.16 ± 5.68 vs.63.89 ± 3.26 at 4 hours,81.16 ± 4.54 vs.176.56 ± 5.45 at 24 hours;TNF-α (ng/L):171.76± 5.60 vs.194.62± 14.13 at 4 hours,328.48 ± 10.79 vs.599.62± 10.20 at 24 hours;D-lactate (mmol/L):0.21 ±0.01 vs.0.24±0.02 at 4 hours,0.24±0.02 vs.0.33±0.04 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,intestinal injury was ameliorated,the Chiu score was significantly lower [1.5 (1.0-5.0) vs.4.5 (3.0-5.0),P ＜ 0.05],and HO-1 positive cells in the intestinal mucosa was remarkably increased.Conclusion UFH can enhance the expression of HO-1 in intestinal mucosa,reduce the release of inflammatory factors,ameliorate the intestinal inflammatory response,and thus play a protective role in intestinal tissue in mice with sepsis.

Drug targets discovery is one of the most important elements in new drug development, and a variety of methods have been developed recently from this point of view. This paper proposed a network-based local and global consistency for cardiovascular genes identification. Results were evaluated through the widely used database HPRD and DrugBank. Results showed that our algorithm can give reasonable candidate targets set. The method in this paper could be an impressive solution for targets searching.
Algorithms ; Cardiovascular Diseases ; genetics ; metabolism ; prevention & control ; Databases, Protein ; Drug Delivery Systems ; methods ; Drug Discovery ; methods ; Gene Regulatory Networks ; Humans ; Models, Theoretical ; Pharmaceutical Preparations ; administration & dosage ; metabolism ; Protein Binding ; Protein Interaction Mapping ; methods ; Proteins ; genetics ; metabolism

Network pharmacology, as a new developmental direction of drug discovery, was generating attention of more and more researchers. The key problem in drug discovery was how to identify the new interactions between drugs and target proteins. Prediction of new interaction was made to find potential targets based on the predicting model constructed by the known drug-protein interactions. According to the deficiencies of existing predicting algorithm based bipartite graph, a supervised learning integration method of bipartite graph was proposed in this paper. Firstly, the bipartite graph network was constructed based on the known interactions between drugs and target proteins. Secondly, the evaluation model for association between drugs and target proteins was created. Thirdly, the model was used to predict the new interactions between drugs and target proteins and confirm the new predicted targets. On the testing dataset, our method performed much better than three other predicting methods. The proposed method integrated chemical space, therapeutic space and genomic space, constructed the interaction network of drugs and target proteins, created the evaluation model and predicted the new interactions with good performance.
Algorithms ; Drug Delivery Systems ; methods ; Drug Discovery ; methods ; Genomics ; methods ; Models, Theoretical ; Pharmaceutical Preparations ; administration & dosage ; metabolism ; Protein Binding ; Protein Interaction Mapping ; methods ; Proteins ; genetics ; metabolism