DNA, Neoplasm ; analysis ; Female ; Flow Cytometry ; Humans ; Hydatidiform Mole ; diagnosis ; genetics ; In Situ Hybridization, Fluorescence ; Microsatellite Repeats ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Pregnancy ; Triploidy ; Uterine Neoplasms ; diagnosis ; genetics

Female ; Humans ; Hypothyroidism ; drug therapy ; Infant, Newborn ; Thyroxine ; poisoning ; therapeutic use

Objective To explore the effects of TNF-α on the expression of IP3 R1 mRNA and protein in human mesangial cells (HMCs) and elucidate the role of protein kinase C (PKC) in this signal pathway.Methods Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effects of TNF-α on IP3R1 mRNA and protein expression.Depletion PKC,the selective inhibitor of PKCα Safingol and inhibitor of PKCδ Rottlerin,overexpression of dominant negative mutant of PKC to examine the mechanism of signal transduction of TNF-α-regulated IP3 R1 in HMCs.PKCα activation was assayed by Western blot.Results TNF-α increased IP3R1 mRNA and protein expression in HMCs,effects that were blocked by prolonged incubted chronic PMA,Safingol and also by domain negative PKCα construct.TNF-α promoted PKCα activation with maximal PKCα phosphorylation that occurred 8 h post-stimulation.Conclusion TNF-α increased the expression of IP3 R1 and this was mediated through the PKCα activation signaling pathways in HMCs.

Objective To explore the effects of TNF-α on the expression of IP33R1 mRNA and protein in human mesangial cells (HMCs) and elucidate the mechnism of TNF-α indnces the IP3R1 expression in the occurrence of hepatorenal syndrome (HRS).Methods HMCs was stimulated by tumor (TNF-α) with 100 ng/mL for different hours (2,4,8,24 hours).The expression change of IP3R1 mRNA and protein were detected by quantitative real-time polymerase chain reaction and immunoblot assay.Several inhibitors including D609,U73122,PP1,Safingol,Rottlerin and non-radioactive PKC assay to examine the mechanism of signal transduction of TNF-α-regulated IP3R1 in HMCs.Results The levels of IP3R1 mRNA at 2 h post-TNF-α exposure were significantly enhanced and reached peak at 8 h in HMCs (P ＜ 0.01),then descened at 24 h (P ＜ 0.01).The levels of IP3R1 protein at 4 h post-TNF-α exposure were obviously increased and reached peak at 24 h post-TNF-α exposure (P ＜ 0.01).Compared with the control group,safingol (PKC-α inhibitor) and D609 (PC-PLC inhibitor) each significantly suppressed TNF-α-induced expression of IP3R1 mRNA (3.30 ± 0.81) vs.(1.95 ± 0.130,P ＜ 0.05 ; (2.10 ± 0.49),P ＜ 0.01 andIP3R1 protein (3.09±0.13) vs.(1.86+0.39),P＜0.01; (1.98±0.02),P＜0.01.TNF-αpromoted autophosphorylation,and hence the activation,of PKC-α with maximal phosphorylation that occurred 8 h post-stimulation measured by non-radioactive PKC assay,and the effect was marked attenuated by pretreated with D609 or safingol.Conclusions TNF-α increased the expression of IP3R1 and this was mediated,at least in part,through the PC-PLC/PKC-α signaling pathways in HMCs.

ObjectiveTo explore the cytogenetic characteristics of multiple myeloma (MM) patients,to evaluate the effect of a long-term culture stimulated by cytokines on cytogenetic study of MM,and to investigate the clinical detection value of RB1 and P53 deletion in interphase plasma cells by using fluorescence in situ hybridization (FISH).MethodsKaryotype analysis was performed in 81 MM patients by using the short-term culture of bone marrow cell and G-banding technique.Among the 81 MM patients,28 patients used two culture methods:one was the short-term culture and the other was to culture cells for 6 days with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) (40 μg/L) and IL-6 (10 μg/L).RB1 and P53 deletion were detected on interphase plasma cells by using FISH in 31 patients.ResultsAmong the 81 patients,75 had enough metaphases for analysis.Among the 75 patients,31 (41.3％)had clonal karyotypic abnormalities including 4 numeric abnormalities,11structural abnormalities and 16 both abnormalities.Among the 28 patients using two culture methods,the clonal karyotypic abnormalities were detected in 6 patients(25.0％ ) in the group of cultured for 24 hours,and 14 patients (51.9％) in 6-day culture group with a significant difference (P =0.026).RB1 deletion and P53 deletion were detected in 10 patients (32.3％ ) and 11 patients(35.5％ ),respectively,with both RB1 and P53 deletions be detected in 5 patients ( 16.1％ ).ConclusionsMore than half of the tested MM patients have both numeric and structural chromosome abnormalities.The karyotype analysis using banding technique is basic cytogenetic study.Extended culture in the presence of IL-6 and GM-CSF could improve the efficiency of cytogenetic analysis to MM.Interphase FISH is a sensitive method of clinical application significance to detect the gene deletion of MM.

OBJECTIVE:To observe the ultra-filtration and the resistance to pollution of5ultra-filtrated membrane in respect of each of the compositions from the Isatis indigotica Fort.water-extraction,in order to produce the Isatis indigotica Fort.-preparation through ultra-filtrated method instead of ethanol-sedimentary method.METHODS:By measuring the quantities of the solid residue and the contents of polysaccharide,arginine and chloroform extraction in the ethanol-sedi?mentary solution and ultra-filtrated solution by gravimetry,TLC-scan and HPLC respectively.RESULTS:The solid residue in the ultra-filtered solution was lower than that in the ethanol-sedimentary solution;The content of polysaccharide in the ultra-filtered solution with polypropylene nitrile membrane and polysulfone-6000membrane were close to that in the ethanol-sedimentary solution.The arginine in the ultra-filtered solution was higher than that in the ethanol-sedimentary solution,but the chloroform extraction in the ultra-filtered solution was lower than that in ethanol-sedimentary solution;The resistance to pollution of polysulfone membrane was the strongest in these ultra-filtrated membrane.CONCLUSION:According to the requirement for isolation of active components,ultra-filtrated method with polysulfone-50000membrane could be used instead of ethanol-sedimentary method for efficient and economical application.

Objective To investigate the effects of curcumin on H2O2-induced the production of nitric oxide(NO) and reactive oxygen species(ROS) in mouse embryo-fibroblast.Methods Macrophage were collected in abdominal cavity of 6-8 weeks Kunming mouse,cultured macrophage(2?108 L-1) were divided into control group and curcumin groups randomly.Macrophage in H2O2 group were added into a single bolus of H2O2(1 mmol?L-1) for 30 min,macrophage in curcumin group were preincubated with different concentration of curcumin for 2 h followed by a 30 min incubation with 1 mmol?L-1 H2O2 and macrophage in control group were added into the same volume(0.1 mL) of 9 g?L-1 so-dium chloride.Immunocytochemistry was used to measure the contents of inducible nitric-oxide synthase(iNOS),NO production was determined by Griess reactive,ROS was determined by DCFH-DA Fluorescence proe.Results The production of NO in control group was little.The production of NO in H2O2 group markly highter than that in control group(P

Objective To explore the distribution characteristics of monosomal karyotype (MK) in elderly patients with acute myeloid leukemia (AML).Methods The karyotype analysis was performed in 123 elderly patients with newly diagnosed AML in our center from Nov 2000 to Feb 2015.We retrospectively studied the distribution characteristics of monosomal karyotype in these patients.Results Among 123 elderly patients with AML,117 patients had enough metaphases chromosomes for analysis.Among the 117 patients,there were 16 cases with good-risk karyotype,54 cases with intermediate-risk karyotype,and 47 cases with adverse-risk karyotype.In the 47 patients with adverse-risk karyotype,43 cases had complex karyotypes (CK).In the 117 elderly AML patients,37 cases (31.6％) had monosomal karyotype (MK),22 AML cases were secondary to myelodysplastic syndrome (MDS-AML),among them 13 cases (55.0％) had MK.In the 95 cases with primary AML,the detection rate of MK was 25.3％ (24 cases).The detection rate of MK+ AML was higher in MDS-AML patients than in de novo AML patients (P=0.000).Among the 37 patients with MK+AML,35 cases had complex karyotypes.30 (81.1％) MK+AML patients had two or more distinct autosomal monosomies and 7 (18.9％) MK+ AML patients had one single autosomal monosomy in the presence of structural abnormalities,and the incidence of autosomal monosomies was higher than that of single autosomal monosomy.The presence of--5 (27.0％),-4 (18.9％),-7 (16.3％) and-6 (13.5％) chromosomes was the most common autosomal monosomy among MK+ AML patients.Conclusions The detection rate of MK is relatively high in elderly AML patients.Two or more distinct autosomal monosomies are more common.The detection rate of MK+AML is higher in patients with MDS-AML than in patients with de novo AML.

Acinetobacter Infections ; microbiology ; Acinetobacter baumannii ; drug effects ; isolation & purification ; Anthracosis ; microbiology ; Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; Fosfomycin ; pharmacology ; Humans ; Imipenem ; pharmacology ; Pneumonia, Bacterial ; microbiology

BACKGROUND:There are many researches on the finite element analysis of total hip arthroplasty, but the biomechanical distribution after hip surface replacement is little reported.OBJECTIVE: To analyze the biomechanical characteristics after hip surface replacement based on three- dimensional finite element analysis.METHODS: The three-dimensional finite element model after metal-to-metal hip surface replacement was established.The load of 350 N paralleling to the gravity line was loaded on the acetabulum. The stress distribution of the prosthesis and femur and the stress of the different regions of the femoral head were observed.RESULTS AND CONCLUSION: (1) The stress value of the acetabular cup was significantly higher than that of the acetabular body ((0.63± 0.34) vs. (1.89±0.67) MPa, P < 0.05). (2) The stress was concentrated on the femoral neck. The femoral head and femoral neck junction appeared with stress occlusion. The stress around the femoral prosthesis stem was (158.24±28.79) MPa, and the stress value of the femoral prosthesis stem was (186.46±22.37) MPa, showing no significant difference (P > 0.05). (3) The stress value showed significant difference among femoral head regions (P <0.05), and the stress of the regions 6 and 8 was significantly higher than that of the other regions (P < 0.05). (4) These results show that after metal-to-metal hip surface replacement, femoral neck stress is concentrated, the stress of the femoral prosthesis is occluded, and the stress at the medial femoral neck as well as the junction of femoral neck prosthesis and bone is larger.