Objective To investigate the synergistic effects of 9-cis-retinoic acid(9-cis-RA) and 8-cl-cAMP on growth inhibition and apoptosis induction in H460 and H292 cell lines of non-small-cell lung cancer(NSCLC). Methods Experimental groups included 9-cis-RA groups(1,5,10 and 20 ?mol/L),8-cl-cAMP groups(5,10,20 and 50 ?mol/L),9-cis-RA(5 and 10 ?mol/L) combined with 8-cl-cAMP(10 ?mol/L) groups and blank control group.The cell growth inhibition rates were detected by trypan blue staining,and the apoptosis of H460 and H292 cells were observed by Hoechst33258 fluorescence microscope,DNA gel electrophoresis and flow cytometer(FCM). Results 9-cis-RA inhibited the growth of H460 cells in a time-and dose-dependent manner,and induced the apoptosis of H460 cells(P

Objective To study the mRNA expression of myelin basic protein (MBP) and myelin oligodendregha glyeoprotein (MOG) in hippocampus of rats following global brain ischemia.Method The four- vessel occlusion animal model in the Sprague-Dawley rats was used in this study.The mRNA expression levels of MBP and MOG in the hippocampus of rats were analyzed by reverse transcription polymerase chain reaction (RT- PCR) at day 2,4,7,14 and 28 days after global brain ischemia.There were eight rats at each time-point and sham operated group.Results The mRNA expression of both MBP and MOG in hippocampus of rats decreased at 2 days after global brain ischemia.The gene expression of myelin gene decreased significantly at 7 days and it reached to the lowest level at 28 days.Compared with sham operated group,the gene expression of MBP and MOG in hippocampus of rats decreased significantly at 7,14 and 28 days after global brain ischemia (P

Objective To make a comparison for the neutrophils prepared either by induction of differentiation of myeloid leuke-mia cell line,or by separation and purification of peripheral blood cells,or by induction of myeloid differentiation of peripheral blood stem cells.Methods NB4 cells were induced differentiation by 1μmol/L all-trans retinoic acid (ATRA)to mature granulo-cytes;neutrophils were separated and purified from peripheral blood by lysis of red blood cells followed by negative selection using magnetic bead-labeled antibodies;hematopoietic stem cells were separated and purified from peripheral blood by Percoll gradient centrifugation followed by negative selection using magnetic bead-labeled antibodies,and were induced to myeloid differentiation by GM-CSF and G-CSF.Morphology and purity of neutrophils prepared by these three methods were studied by means of MGG stai-ning.CD18 protein expression and subcellular distribution were studied by means of immunofluorescence staining.Results Purity of neutrophil was above 40% by induction of differentiation of NB4 cells,and was about 90% if purified from peripheral blood,and was above 70% if induced by myeloid differentiation of peripheral blood stem cells.There was no obvious difference for CD18 ex-pression in neutrophils prepared by these three methods,and staining of CD18 had a dotted pattern distributed in these cells.Con-clusion Peripheral blood neutrophils prepared by lysis of red blood cells followed by negative selection using magnetic bead-labeled antibodies are with high purity and viability which is suitable for immediate test of neutrophils from fresh blood.Neutrophils pre-pared by myeloid differentiation of hematopoietic stem cell are with high viability and last for days,which can be used in long test for neutrphils.

OBJECTIVETo explore the effect of miR-542-3p in malignant transformation of human bronchial epithelial cells (16HBE) induced by anti-benzo(a)pyrene-7,8-diol-9,10-epoxide (anti-BPDE).

METHODSThe relative expression level of mature miR-542-3p in transformed cells (16HBE-T) and untransformed control cells (16HBE-N) was measured by real-time quantitative polymerase chain reaction (qRT-PCR). miRNA mimic was transiently transfected into 16HBE-T to change the expression level of miR-542-3p, and then the influenced changes of cell proliferation, cell cycle, apoptosis, and soft agar colony formation rate and the migration of transfected cells were analyzed.

RESULTSBefore transfection, the expression level of mature miR-542-3p in 16HBE-T was lower (39.08 ± 6.95)% than it in 16HBE-N (t = 15.18, P < 0.05). In comparison with the 16HBE-T group, the expression level of miR-542-3p in miR-542-3p mimic-transfected group was (5.23 ± 0.55) fold (t = 17.37, P < 0.05) after transfection. Cell proliferation of mimic-transfected group was decreased to (62.06 ± 5.61)% (t = -17.28, P < 0.05), percentage of cells in G(0)/G(1) phase up to (74.76 ± 4.86)% (t = 4.53, P < 0.05), rate of colony formation degrade to (5.87 ± 0.67)% (t = -6.66, P < 0.05), coverage areas ratio decreased to (0.31 ± 0.08) (t = -6.78, P < 0.05). There was no change with apoptosis.

CONCLUSIONOur studies showed that miR-542-3p played the role as a tumor suppressor, which led to a significant decrease in the proliferation capacity and degree of malignancy. These findings suggest aberrantly down-regulated miR-542-3p may be one critical factor that contributes to malignant transformation of 16HBE induced by anti-BPDE.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; adverse effects ; Bronchi ; cytology ; Cell Transformation, Neoplastic ; drug effects ; genetics ; metabolism ; Epithelial Cells ; cytology ; drug effects ; Humans ; MicroRNAs ; genetics ; Transfection

OBJECTIVETo study the chemical constituents of Polyporus ellissi.

METHODSilica gel column chromatography was applied for the isolation and purification of the constituents. The structures were established by means of spectroscopic and chemical data.

RESULTSix compounds were obtained and identified as cerebroside B (I), cerebroside D (II), ergosterol peroxide (III), 9(11)-dehydroergosterol peroxide (IV), mannitol (V) and palmitate-1-glycerol (VI).

CONCLUSIONCompounds (I) and (II) were isolated from the genus Polyporus for the first time.

Cerebrosides ; chemistry ; isolation & purification ; Mannitol ; chemistry ; isolation & purification ; Molecular Structure ; Polyporaceae ; chemistry

OBJECTIVETo establish a rapid and simple method to detect Y chromosome microdeletions directly using whole blood as starting material for multiplex-PCR.

METHODSUsing a self-prepared pHpH-Bufferq, multiplex-PCR amplification was directly carried out from the anticoagulant whole blood sample without DNA extraction step. Twelve sequence tagged sites (STS), namely SY84, SY86, SY127, SY134, SY124, SY132, SY152, SY157, SY239, SY242, SY254 and SY255, in AZFa, AZFb, and AZFc gene regions were detected in 5 different tubes. In order to ensure the validity of the experiments, sex-determining region Y (SRY) and X-linked or Y-linked zinc finger gene (ZFX/Y) were used as internal controls. Furthermore, conventional PCR using genomic DNA extracted from each blood sample was performed in parallel for evaluating the accuracy of the experiments.

RESULTSA total of 156 male samples were detected, and a normal male sample and a female sample were used as a positive and a negative control respectively. The results showed that 144 samples had no deletion; one sample was AZF-deleted; one sample was AZFb-deleted; seven samples were AZFc-deleted; one sample was both AZFb- and AZFc- deleted; and two samples were all AZFa-, AZFb- and AZFc- deleted. The observed results from two kinds of starting material (whole blood and purified DNA) are completely consistent.

CONCLUSIONIn our method, PCR amplification was directly carried out from whole blood without any DNA extraction step. So it has the advantages of low cost, simple process, time-saving operation and less cross-contamination. The whole process can be completed within 2 hours. Thus the efficiency of clinical detection is improved greatly.

Azoospermia ; genetics ; Cells, Cultured ; Chromosome Deletion ; Chromosomes, Human, Y ; Female ; Humans ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Sex Chromosome Aberrations

OBJECTIVETo investigate the effect of farnesoid-X-receptor (FXR) antagonist Z-guggulsterone in an in vivo high-fat fed apolipoprotein E knockout (ApoE(-/-)) mice model of myocardial ischemia/reperfusion (I/R).

METHODSMale ApoE(-/-) mice were randomly divided into three groups: standard ApoE(-/-) group (fed with standard mouse diet for 12 weeks before myocardial I/R procedure, n = 18), high-fat ApoE(-/-) group (fed with high-fat mouse diet for 12 weeks before myocardial I/R procedure, n = 22), and high-fat ApoE(-/-) + FXR antagonist group(fed with high-fat mouse diet for 12 weeks and received FXR antagonist Z-Guggulsterone 30 minutes before myocardial I/R procedure, n = 17). The expression of FXR was detected by real-time quantitative-PCR. Myocardial infarct size was determined by Evans blue/TTC double staining methods. Myocardial apoptosis was determined by in situ TUNEL technique. Markers of the mitochondrial-mediated apoptotic pathway (cytochrome c release, caspase-9 activity, and BAX and BCL-2 levels), endoplasmic reticulum stress apoptotic pathway (caspase-12 activity and CHOP level), and death receptor apoptotic pathway (caspase-8 activity, and Fas and FasL levels) were also measured.

RESULTFXR expression (3.7-fold higher, P < 0.01), myocardial infarct size [(62.1 ± 7.0)% vs. (33.8 ± 5.8)%, P < 0.01] and myocardial apoptosis index[ (36.8 ± 5.7)% vs. (17.2 ± 3.8)%, P < 0.01]were all significantly higher in high-fat ApoE(-/-) group than those in standard ApoE(-/-) group. Compared with high-fat ApoE(-/-) group, myocardial infarct size [(24.4 ± 4.7)% vs. (62.1 ± 7.0)%, P < 0.01] and myocardial apoptosis index [(13.8 ± 2.7)% vs. (36.8 ± 5.7)%, P < 0.01] were significantly reduced in high-fat ApoE(-/-) + FXR antagonist group. Moreover, levels of mitochondrial-mediated apoptotic pathway markers (cytochrome c release, caspase-9 activity, and BAX/BCL-2 levels) and endoplasmic reticulum stress apoptotic pathway markers (caspase-12 activity and CHOP level) were significantly lower in high-fat ApoE(-/-) + FXR antagonist group than those in high-fat ApoE(-/-) group (all P < 0.01). Levels of death receptor apoptotic pathway markers (caspase-8 activity, and Fas and FasL levels) were similar between high-fat ApoE(-/-) group and high-fat ApoE(-/-) + FXR antagonist group.

CONCLUSIONFXR antagonist alleviates myocardial reperfusion injury in cholesterol-fed ApoE(-/-) mice via inhibition of the mitochondrial-mediated and endoplasmic-reticulum stress pathway.

Animals ; Apolipoproteins E ; genetics ; Apoptosis ; drug effects ; Caspase 9 ; metabolism ; Cholesterol, Dietary ; administration & dosage ; Cytochromes c ; metabolism ; Disease Models, Animal ; Endoplasmic Reticulum Stress ; Male ; Mice ; Mice, Knockout ; Myocardial Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Pregnenediones ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Receptors, Cytoplasmic and Nuclear ; antagonists & inhibitors ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo study the gene expression of Notch1 and Jagged1 in children with acute leukemia (AL) and their possible roles in the pathogenesis of AL.

METHODSMononuclear cells from bone marrow or peripheral blood of 47 children with AL and 20 controls (normal children or children with nonmalignant hematologic disease) were collected from February 2009 to July 2011. A two-step method to semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the gene expression of Notch1 and Jagged1. Of the 47 children with AL, there were 26 cases of B-ALL, 6 cases of T-ALL and 15 cases of AML.

RESULTSThe positive expression rate of Notch1 in the ALL and AML groups was higher than in the control group (P<0.05). The expression level of Notch1 in T-ALL children was higher than in B-ALL children (P<0.01). The positive expression rate of Jagged1 in the ALL and AML groups was not significantly different from the control group, however, the expression level of Jagged1 in the ALL and AML groups was higher than in the control group (P<0.05).

CONCLUSIONSThere are significant differences in the gene expression of Notch1 between children with different types of ALL, and a higher expression of Notch1 relates to T-ALL. The activation of Notch1 signal is common in children with AL. The abnormal gene expression of Notch1 in children with AML shows the role of Notch1 in AML. The gene expression of Jagged1 in children with ALL or AML is abnormal, and this needs to be confirmed by further research.

Acute Disease ; Adolescent ; Calcium-Binding Proteins ; genetics ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Infant ; Intercellular Signaling Peptides and Proteins ; genetics ; Jagged-1 Protein ; Leukemia ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Leukemia-Lymphoma, Adult T-Cell ; metabolism ; Male ; Membrane Proteins ; genetics ; Receptor, Notch1 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serrate-Jagged Proteins ; Signal Transduction

OBJECTIVETo explore the impact of glycogen synthase kinase-3β (GSK-3β) on Wnt and NF-κB pathways in a rat model of diabetic nephropathy (DN).

METHODSSD rats were randomly divided into normal control group (NC), DN model group (DM) and GSK-3β inhibitor group (DI). Blood glucose and 24-hour urine protein were monitored in three groups. Renal tissue samples were stained by HE. The expression of GSK-3β and NF-κB proteins was studied by immunohistochemistry. GSK-3β and NF-κB mRNAs were detected by RT-qPCR.

RESULTSTen weeks after STZ injection, the level of blood glucose increased significantly in DM group [(23.2±5.4) mmol/L] and DI group [(25.0±4.0) mmol/L], compared with NC group, and the level of 24-hour urinary protein increased significantly in DM group [(185.2±35.6) g/24 h] and DI group [(179.6±44.7) g/24 h], compared with NC group. Two weeks after LiCl injection, the level of blood glucose and 24-hour urinary protein decreased in DI group (17.6±2.1) mmol/L, (106.9±30.0) g/24 h], compared with DM Group. Compared with NC group, pathological changes of the kidney of DM group aggravated along with increased mRNA and protein expression of GSK-3β and NF-κB. But the pathological changes of the kidney in DI group alleviated along with declined mRNA and protein expression of GSK-3β and NF-κB as compared with DM group (all P<0.05).

CONCLUSIONSNF-κB protein expression positively correlates with the GSK3β expression. Wnt and NF-κB signal pathways play an important role in the development of diabetic nephropathy.

Animals ; Diabetic Nephropathies ; metabolism ; Disease Models, Animal ; Glycogen Synthase Kinase 3 ; antagonists & inhibitors ; metabolism ; Glycogen Synthase Kinase 3 beta ; Kidney ; pathology ; NF-kappa B ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Wnt Signaling Pathway

OBJECTIVETo probe the diagnostic value of the infrared radiation spectrum of acupoint for ulcerative colitis (UC).

METHODSA high sensitivity PHE 201 infrared spectrum instrument was used to determine the infrared radiation spectrum of Hegu (LI 4) and Shangjuxu (ST 37) in 34 cases of UC.

RESULTSOf 59 waves detected, there were significant differences in infrared radiation intensity of 28 different waves between the healthy people and the patients with UC in right Hegu (LI 4) (P < 0.05 or P < 0.01) and 13 waves in left Hegu (LI 4) (P < 0.05); there were significant differences in 16 different waves in right Shangjuxu (ST 37) (P < 0.05 or P < 0.01) and in 17 waves in left Shangjuxu (ST 37) (P < 0.05 or P < 0.01); there was a significant difference in 18 waves between right and left Hegu (LI 4) of the patients (P < 0.05 or P < 0.01) and 7 waves between right and left Hegu (LI 4) of the healthy people (P < 0.05). There was a significant difference in 4 waves between right and left Shangjuxu (ST 37) of the patients and one wave between right and left Shangjuxu (ST 37) of the healthy people (P < 0.01).

CONCLUSIONBoth Hegu (LI 4) and Shangjuxu (ST 37) show changes of infrared radiation spectrum when the intestine gets lesion, and Hegu (LI 4) can better show the change.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Colitis, Ulcerative ; therapy ; Female ; Humans ; Infrared Rays ; Intestine, Large ; anatomy & histology ; Male ; Middle Aged