Animals ; Genes, p53 ; Genetic Therapy ; Humans ; Neoplasms ; genetics ; metabolism ; therapy ; Proto-Oncogene Proteins c-mdm2 ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

0.05).The degreeⅢ～Ⅳthrombocytopenia was more common in group A than in group B,but the degreeⅢ～Ⅳhypolekocytosis and phlebitis was more serious in group B.Conclusion NC and GC for treating refractory metastatic breast cancer have a high response rate and tolerable side effects.

Objective To investigate the association between leukoaraiosis (LA) and early neurological deterioration (END) in patients with acute ischemic stroke.Methods Clinical data of 328 patients with acute ischemic stroke admitted in the hospital from January 2013 to January 2016 were retrospectively reviewed.According to the changes of National Institute of Health Stroke Scale (NIHSS) scores within 72 h after admission,88 patients (26.8%) were identified as END.Clinical manifestations,laboratory tests and radiographic findings were compared between END group and non-END group.ResultsUnivariate analysis indicated that age [(74.6±11.0) vs.(70.7±11.8) years,t=2.67,P=0.01],female sex [51.1% (45/ 88) vs.38.8%(93/240),χ2=4.05,P=0.04],initial NIHSS [M(Q1,Q3) 6(3,9) vs.3 (2,6),χ2=-4.38,P=0.00],systolic blood pressure [(155±28) vs.(149±20) mmHg(1 mmHg=0.133 kPa),t=2.04,P=0.04],responsible artery occlusion [18.2% (16/88) vs.8.3%(20/240),χ2=6.39,P=0.01],white cell count [(7.8±2.7) 109 vs.(7.1±2.2) 109,t=2.32,P=0.02],fasting blood glucose [(7.2±2.6) vs.(6.6±2.4) mmol/L,t=2.00,P<0.05] and C-reactive protein level [(24.5±27.1) vs.(14.6±23.2) g/L,t=3.25,P=0.00] were significantly different between END group and non-END group.After adjustment of confounding factors,LA in periventricular with Fazekas grade 2 (OR=2.309,95%CI: 1.070-4.984,P=0.03) and Fazekas grade 3 (OR=2.861,95%CI: 1.214-6.742,P=0.02) and LA in centrum semiovale with Fazekas grade 3 (OR=3.047,95%CI: 1.244-7.461,P=0.02) were independently associated with END.Conclusion Leukoaraiosis in periventricular group and centrum semiovale are associated with early neurological deterioration in patients with acute ischemic stroke.

Objective To investigate the renoprotective effect of transforming growth factor beta activator kinase 1 (TAK1) inhibitor 5Z-7-oxozeaenol (OZ) in diabetic db/db mice and the mechanism.Methods Twenty-four male db/db mice were randomly divided into two groups:db/db mice (db/db,n=12) and db/db mice with 5Z-7-oxozeaenol treatment (db/db+OZ,n=12).Another group of wild type mice (n=12) was held as the control group.OZ 2 mg/kg was administrated by intraperitoneal injection every other day.At week 8 and 12 after 5Z-7-oxozeaenol treatment,blood glucose (BG),body weight (BW),kidney weight (KW) and urinary albumin excretion rate (UAER) were evaluated.Kidney pathological lesions were detected by light and electron microscopy.NF-κB p65,monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-ot (TNF-o) were detected by immunohistochemistry.Western blotting was used to detect p-TAK1,TAB1,p-p38MAPK and IL-1β expression,while ICAM-1 and MCP-1 mRNA levels were evaluated by real-time PCR.Results Compared with control group,the levels of BG,BW,KW and UAER were higher (P ＜ 0.01) in db/db mice group,while BW,KW and UAER levels were significantly decreased in db/db + OZ group compared with that in db/db mice group (P ＜ 0.05).In week 8 and 12 db/db mice,glomerular volume and extracellular matrix were increased,while pathological lesions in kidney tissue were positively improved by TAK1 inhibitor.Immunohistochemistry showed that NF-κB p65,MCP-1 and TNF-α expression levels were apparently increased in db/db mice group compared with that in control group (P ＜ 0.05) and were significantly inhibited by TAK1 inhibitor (P ＜ 0.05).Western blotting showed that p-TAK1,TAB1,p-p38MAPK and IL-1β expression levels were higher in db/db mice group than that in control group (P ＜ 0.05) and lower in db/db+ OZ group than that in db/db mice group (P ＜ 0.05).Moreover,real-time PCR showed that the expressions of ICAM-1 and MCP-1 mRNA were higher in db/db mice group than that in control group and lower in db/db+OZ group than that in db/db mice group (P ＜0.05).Conclusions TAK1 Inhibitor can down-regulate MAPK and NF-κB pathway to restrain the reaction of inflammation and alleviate kidney injury in diabetic db/db mice.

Objective To investigate the role of transforming growth factor-β activated kinase-1 (TAK1) signaling pathway in the activation of bone marrow derived macrophages (BMDM) induced by high glucose.Methods Purity of mouse BMDM was detected by flow cytometry.The mice macrophages cultured in vitro were stimulated by high glucose and treated with TAK1 specific inhibitor 5Z-7-oxozeaenol.Cells were divided into normal control group (RPMI 1640),osmolality control group (25 mmol/L mannitol),high glucose group (33 mmol/L D-glucose) and inhibitor group (33 mmol/L D-glucose+300 nmol/L 5Z-7-oxozeaenol).Immunocytochemistry and flow cytometry were used to detect macrophage subtype.The expression of monocyte chemotactic protein-1 (MCP-1) and tumor necrosis Factor-α (TNF-α) mRNA were determined by real time PCR.Expressions of p-TAK1,TAK1 binding protein (TAB1),p-JNK,p-p38 MAPK and NF-κB p65 proteins were analyzed by Western blotting.Results The purity of BMDM was about 99.36％.Compared with normal control group,high glucose group had increased percentage of M1 macrophages,increased expression of MCP-1 and TNF-α mRNA (all P ＜ 0.05).Moreover,p-TAK1,TAB1,p-JNK,p-p38 MAPK and NF-κB p65 proteins expression also increased significantly in high glucose group (all P ＜ 0.05).After treatment with inhibitor 5Z-7-oxozeaenol,the effects induced by high glucose were inhibited (P ＜ 0.05).Conclusions High glucose can induce M1 macrophage activation and expression of inflammatory cytokine of BMDM,which can be inhibited 5Z-7-oxozeaenol through inhibiting TAK1/MAPK and TAK1/NF-κB pathway.

BACKGROUND:Studiesin vitro have suggested that icarin can attenuate lipopolysaccharide (LPS)-induced acute pneumonia. Is the anti-inflammatory effect of icarin stil valid in the presence of wear particles? OBJECTIVE:With studiesin vivo andin vitro, to investigate the regulatory effect of icarrin on titanium particle-induced inflammatory reaction. METHODS:(1) Studiesin vivo: Eighty male C57BL/6 mice aged 6-8 weeks were randomly divided into four groups: control group, icarin group, titanium particle group, and titanium particle+icarin group. Mice in the titanium particle group and titanium particle+icarin group received surgical procedure, and sterile and endotoxin-free titanium particles were implanted on the calvaria surfaces to induce inflammatory reaction. Mice in the control group and icarin group received the same surgery, but no wear particles were implanted. Then icarin was given oraly to mice in the titanium particle group and titanium particle+ icarin group with a dose of 200 mg/kg per day for 2 weeks from the day of modeling. Mice in the control group and icarin group were given oraly the same dose of placebo. Two weeks later, tumor necrosis factor-α and interleukin-1β at protein and mRNA levels were respectively detected with enzyme-linked immunohistochemical (ELISA) and quantitative real time reverse transcription PCR (qRT-PCR) analysis. (2) Studiesin vitro: Mouse monocyte/macrophage RAW264.7 cels were cultured with different conditioned media: control group, nuclear factor receptor ligand кB (RANKL); icarin group, RANKL+icarin; titanium particle group, RANKL+titanium particles; titanium particle+icarrin group, RANKL+icarin+titanium particles. Titanium particles stimulated RAW264.7 cels were co-cultured with RANKL and icarin for 72 hours. Tumor necrosis factor-α and interleukin-1β at protein and mRNA levels in the supernatant were detected with ELISA analysis and qRT-PCR, respectively. RESULTS AND CONCLUSION: (1) Resultsin vivo: icarin treatment obviously decreased titanium particle-induced inflammatory cellinfiltration and made the thickness of periosteum thinner, down-regulated tumor necrosis factor-α and interleukin-1β expressions at protein and mRNA levels. (2) Results in vitro: when RAW264.7 cels were stimulated with titanium particles for 72 hours, tumor necrosis factor-α and interleukin-1β expressions at protein and mRNA levels in culture media increased obviously; when icarin was administrated, tumor necrosis factor-α and interleukin-1βexpressions at protein and mRNA levels down-regulated significantly. These results suggest icarin can obviously suppress titanium particle-induced inflammatory reactionin vivo andin vitro.

Objective To observe the therapeutic efficacy of electrical stimulation and balloon dilatation in treating cricopharyngeal achalasia after a brainstem stroke.Methods Thirty dysphagia patients with cricopharyngeal achalasia after a brainstem stroke were randomly divided into an experimental group and a control group, each of 15.The experimental group was given real-time electrical stimulation and balloon dilatation, while the control group was treated using common electrical stimulation and balloon dilatation.Results Twenty-six patients in the 2 groups returned to oral feeding after treatment.Videofluoroscopy revealed that the cricopharyngeal sphincter had relaxed and the food passed successfully when swallowing.No aspiration was observed.There was no significant difference in swallowing between the two groups, but the average treatment time, days of treatment and cost of therapy in the experimental group were significantly less than in the control group.Conclusions Either real-time electrical stimulation or common electrical stimulation combined with balloon dilatation can treat dysphagia effectively, but the former can shorten the course of treatment and lower its cost.

Aim We used bone marrow-derived macro-phages ( BMMs ) , to explore the mechanism of macro-phage activation and the effect of TGF-β activated ki-nase-1 ( TAK1 ) inhibitor 5 Z-7-oxozeaenol on it under AGEs conditions. Methods The BMMs were obtained from C57 mice, and purity of BMMs was detected by flow cytometry. Cell viability was tested after treatment with different concentrations of TAK1 inhibitors. Laser confocal microscopy was used to detect macrophage M1 subtype . Flow cytometry was used to analyse the macro-phage activated by AGEs. TNF-α and MCP-1 mRNA levels were evaluated by qRT-PCR. Western blot was used to detect the expression levels of TAK1 signal pathway protein. Results AGEs stimulation could in-crese the activity of M1 macrophages,and 5Z-7-oxoze-aenol could inhibit the differentiation of BMMs. Com-pared with control group, AGEs increased the expres-sion of MCP-1 and TNF-α mRNA(P<0. 01). p-TAK1, TAB1,p-JNK,p-p38MAPK and NF-κBp65 proteins ex-pression also increased significantly ( P <0. 05 ) . After treatment with inhibitor, transcription levels of MCP-1 and TNF-α decreased significantly ( P < 0. 05 , P <0. 01 ) . 5 Z-7-oxozeaenol treatment downregulated the expression of p-TAK1,TAB1,p-JNK,p-p38MAPK and
NF-κBp65 proteins ( P <0. 05 ) . Conclusions AGEs can induce BMMs to M1 phenotypic polarization. 5Z-7-oxozeaenol reduces the expression of inflammatory cyto-kine via inhibiting TAK1/MAPKs, MAPKs/NF-κB pathways.

200 citrinin mutants were screened with the inhibition zone method from the transformants library of Monascus ruber M-7 by Agrobacterium-mediate DNA transfer, which contains more than 5,000 transformants.Then 53 mutants, whose citrinin contents ranged from 0.04?g/g to 154.57?g/g in the red fermented rice (RFR), were achieved by high performance liquid chromatography (HPLC).Color values of RFR prepared by these mutants were also detected.The results showed that there was a positive correlation between the citrinin content and the color value among the mutants.These results provide materials and research bases for ferrther studying the relationship between the production of citrinin and pigment of Monascus ruber at molecular level.

According to DNA sequences of the invA gene of Salmonella spp.,the phoA gene of Escherichia coli and the nuc gene of Staphylococcus aureus,three pairs of oligonucleotide primers were designed and synthesized to amplify the special DNA sequences by multiplex PCR. Moreover,the reaction conditions of multiplex PCR were optimized. The results showed the multiplex PCR using the three pairs of primers produced specific amplicons of expected sizes,284bp for Salmonella spp.,622bp for Escherichia coli,484bp for Staphylococcus aureus. The optimized reaction conditions followed as the concentration of primer 40nmol/L for Salmonella spp.,40nmol/L for Escherichia coli,80nmol/L for Staphylococcus aureus,2.4mmol/L Mg 2+ ,200?mol/L dNTP,1.5U Taq DNA polymerase,anneal temperature from 55.0℃ to 57.4℃. Under the condition,the detection limits for DNA template were 10.2pg,10.2pg and 102.0pg for Salmonella spp.,Escherichia coli and Staphylococcus aureus,respectively. The whole process could be completed within 4h. The multiplex PCR assay was a specific,sensitive,rapid and reliable method for detecting Salmonella spp.,Escherichia coli and Staphylococcus aureus,which establish important foundation for simultaneous detection for these three bacteria in food.