Objective To investigate the relationship between endothelial damage and p120-catenin (p120-ctn) in a model of paraquat intoxication,and the modulatory effect of mangiferin on p120-ctn.Methods Human umbilical vein endothelial cells (HUVECs) were cultured in two compartment spreading apparatus in vitro.The endothelial cells were divided into three groups:control group (cultured in DMEM with 10％ fetal bovine serum),paraquat group (paraquat was added to the medium with final concentration of 0.05 μmol/L) and mangiferin group (cultured in medium with addition of paraquat for 30 minutes,then mangiferin was added in a final concentration of 20 μmol/L).The cellular permeability at 6,12,24,48,72 hours after culture in the three groups was measured.The expressions of p120-ctn 1A,p120-ctn 3A mRNA and p120-ctn protein were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot analysis.The distribution of p120-ctn protein was observed by immunofluorescence.Results Compared with control group,cellular permeability in paraquat and mangiferin groups were increased with prolongation of time,and peaked at 72 hours [(29.86 ± 3.98)％,(24.39 ± 2.79)％ vs.(11.71 ± 1.67)％,both P＜0.05].The cellular permeability was significantly lower in mangiferin group than that in paraquat group at different time points (all P＜0.05).At 6 hours after intoxication,the expressions of p120-ctn 1A,p 120-ctn 3A mRNA (gray value) and p 120-ctn protein (gray value) were significantly lower in paraquat group than those in control group (p120-ctn 1A mRNA:0.150 ± 0.024 vs.0.433 ± 0.024,p120-ctn 3A mRNA:0.316 ± 0.043 vs.0.701 ±0.020,p120-ctn protein:0.485 ±0.031 vs.0.763 ±0.038,all P＜0.01).The expressions of p120-ctn 1A,p120-ctn 3A mRNA and p120-ctn protein were significantly higher in mangiferin group than those in paraquat group from 6 hours on (p120-ctn 1A mRNA:0.281 ± 0.021 vs.0.150 ± 0.024,p120-ctn 3A mRNA:0.602 ± 0.042 vs.0.316 ± 0.043,p120-ctn protein:0.675 ± 0.031 vs.0.485 ± 0.031,all P＜0.01),and they were gradually increased with prolongation of time,and peaked at 72 hours (p120-ctn 1A mRNA:1.376 ±0.128 vs.0.150 ± 0.024,p120-ctn 3A mRNA:1.251 ± 0.059 vs.0.316 ± 0.043,p120-ctn protein:0.844 ± 0.050 vs.0.485 ± 0.031,all P＜ 0.01).Under upright fluorescence microscope,p120-ctn was mainly distributed in the cell membrane in control group,with a slight expression in cytoplasm,and no expression in the nuclei.With prolongation of time,p120-ctn expression in the cell membrane was gradually decreased in paraquat group,while it was increased in the cytoplasm and nuclei,with blurring of cell membrane and widening of cellular gap.p120-ctn expression was improved on the cell membrane in mangiferin group at corresponding time points,with decreased in expression in nuclei and cytoplasm.Conclusion The p120-ctn protein plays an important role in the enhancement of endothelial permeability in paraquat intoxication,and mangiferin may attenuate endothelial injury in paraquat intoxication possibly through modulation of p 120-ctn protein.

The strain F12-11-1-2 was isolated from the East China Sea,which had antimitosis activity using Pyricularia oryzae mode.Ac- cording to phenotypical study,salt-aggregation test and 16S rDNA sequence analysis,the strain F12-11-1-2 has been identified to be Bacillus subtilis.

Objective:To express Tumstatin_(183-230)-TRAIL fusion protein and to observe its biological functions.Methods: SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin_(183-230)and TNF-related apoptosis-inducing ligand (TRAIL_(114-281)).An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c_2;the vector was used to transfect E.coli BL21(DE3)and expression of MBP-Tu-T fusion protein was induced by IPTG.Amylose Resin columns were employed to purify the fusion protein.The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation,standard tumor cell cytotoxic assay,in vitro tube formation inhibition,and electron microscopic observation(apoptosis).Results:The expression rate of MBP-Tu-T fusion protein in E.coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation(IC_(50)12.5?g/ml),induced apoptosis of pancreatic cancer cells,and inhibited tube formation.Conclusion:Constructed MBP-Tu-T fusion protein is bifunctional,which lays a solid foundation for further investigation of antitumor effect of Tumstatin_(183-230)-TRAIL in vivo.

Background Following the accelerated speed of population aging in China,the incidence of cataract is rising gradually.Researches indicated that senescenee marker protein-30 ( SMP-30 ) is closely associated with the occurring and developing of cataract. Objective This study was to investigate the expression of SMP-30 in human lens epithelial cells(LECs) of age-related cataract and study the relationship between SMP-30 and apoptosis.Methods Capsulotomy was performed on 80 eyes of 59 patients with simple cortex age-related cataract and 70 eyes of 53 age-matched patients with nucleus age-related cataract.The anterior capsular specimens were obtained by circularly capsulorhexis during the operation.Expressions of the SMP-30 protein and mRNA in the LECs of two types of cataract were detected using immunochemistry and real-time PCR respectively.Apoptosis of the LECs was assayed by TUNEL.The differences of expression of SMP-30 and apoptosis were compared between the two types of cataract.Results Immunochemistry showed that SMP-30 was expressed in cytoplasm of LECs.The expression intensity of SMP-30 was higher in the center zone compared with periphery zone.The apoptosis rate of LECs was significantly higher in the center of the anterior capsule than the periphery in both two types of cataract ( nucleus cataract:19.34％±0.11％ vs 8.32 ％ ± 0.57 ％,P =0.025 ; cortex cataract:42.07 ％ ± 0.86 ％ vs 13.55 ％ ± 0.64 ％,P =0.010 ).The expression amount of SMP-30 mRNA was lower at the periphery than the center of lens in both two types of cataract (nucleus cataract:45.21±2.79 vs 76.42±11.21,P=0.042 ;cortex cataract:108.32±4.32 vs 206.34±15.67,P=0.037 ),and that of nucleus cataract was significantly lower than cortex cataract (60.02±9.08 vs 157.33 ± 13.01,P =0.034),and the apoptosis rate of LECs was declined in the nucleus cataract group compared with the cortex cataract group ( 14.05％ ±0.22％ vs 27.70％ ±0.81％,P =0.007 ). Conclusions LECs apoptosis exists in age-related cataract.SMP-30 probably plays an important role in the formation of cataract.

OBJECTIVETo evaluate the role of Jiangzhi Xiaoban Tablet (JXT) in improving heartfunction of coronary heart disease (CHD) patients by tissue Doppler imaging (TDI) and speckle trackingimaging (STI) technology.

METHODSRecruited were 60 inpatients with confirmed CHD by coronary angiography at First Affiliated Hospital, Hunan University of Traditional Chinese Medicine from October 2013to November 2014. They were assigned to the treatment group (group A) and the control group (groupB) according to random digit table, 30 cases in each group. Patients in group A took JXT, 0.45 g/tablet,4 tablets each time, 3 times per day, while those in group B took Simvastatin Tablet, 20 mg/tablet, 1 tablet each time, once per evening. The therapeutic course for all was 8 weeks. The long axis view of theheart of 18 segments STI Peak strain LS and TDI peak systolic Sa parameters were performed in all patients before and after treatment.

RESULTSBefore treatment segments of STI strain LS and TDI longitudinal peak systolic peak Sa were not statistically different between the two groups (P > 0.05). Each segment of STI peak longitudinal strain LS and TDI peak systolic Sa in the two groups were higher after treatment than before treatment (P < 0.05). After treatment each segment of STI parameters of LS and eachTDI segment parameters of Sa were significantly lower in group B than in group A (P < 0.01).

CONCLUSIONJXT could improve heart function of CHD patients to different degrees, and its curative effect was betterthan that of routine Western medicine (Simvastatin Tablets) treatment.

Coronary Artery Disease ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Echocardiography, Doppler ; Heart ; drug effects ; Humans ; Simvastatin ; therapeutic use ; Tablets

OBJECTIVETo investigate the role of fascin, an actin bundling protein, in the development and progression of human esophageal squamous cell carcinoma (ESCC), and explore its association with the clinicopathologic characteristics and 5-year survival of the patients.

METHODSUsing tissue array and immunohistochemistry, the expression of fascin was determined in 241 ESCC tissues and the corresponding normal esophageal mucosal tissues.

RESULTSESCC tissues showed a significantly higher overexpression rate of fascin than the corresponding normal esophageal mucosal tissues (68.9% vs 15.5%, P<0.05). The overexpression of fascin was correlated to lymph node metastasis and TMN stage, but not to the patients' age, gender, tumor differentiation and general classification. Survival analysis showed that abnormal expression of fascin was associated with the 5-year survival rate of patients with ESCC.

CONCLUSIONSThe abnormal expression of fascin may play an important role in the progression of ESCC, and detection of fascin expression may have important prognostic values.

Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Carrier Proteins ; metabolism ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Microfilament Proteins ; metabolism ; Middle Aged ; Prognosis

OBJECTIVE: To profile urinary metabolite variations from 1, 2-dimethylhydrazine (DMH)-induced precancerous colon rats, Jinfu Kang treated rats and healthy controls.

METHODWe used ethyl chloroformate derivatization and gas chromatography-mass spectrometry (GC-MS) based metabonomic method to analyze rat urines.

RESULTThe time-dependent variations of metabolite profile showed a progressive deviation of the metabolism in the model group from the initial pattern over time and a systemic recovery of the metabolism in the treatment group, which is consistent with the histological results. The in-depth analysis indicated that the disorder of tricarboxylic acid cycle (TCA), tryptophan metabolism, polyamine metabolism and gut flora structure were associated with DMH intervention.

CONCLUSIONMetabolic study revealed that Jinfu Kang can effectively reverse metabolic departures in DMH-induced precancerous colon rat, which is consistent with pathological results.

Animals ; Colonic Neoplasms ; chemically induced ; pathology ; Colonic Polyps ; chemically induced ; drug therapy ; urine ; Dimethylhydrazines ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Gas Chromatography-Mass Spectrometry ; Male ; Rats ; Rats, Wistar

BACKGROUNDExperimental evidence indicates that cyclooxygenase-2 (COX-2) plays a critical role in blastocyst implantation; however, little is known of the role of COX-2 in unexplained recurrent spontaneous abortion (URSA).

METHODSWe evaluated the expression level and potential signaling pathway of COX-2 in 30 cases of URSA who were excluded the abnormality of chromosomes, anatomy, endocrine, infectious, autoimmune diseases and in 30 normal pregnancies.

RESULTSThe mRNA and the protein expression level of COX-2 in the URSA group (-0.238 +/- 0.848, 0.368 +/- 0.089, respectively) were significantly lower than that in the control group (1.943 +/- 3.845, 1.046 +/- 0.108, respectively) (both, P < 0.01). The expression of prostaglandins PGF(2a), PGD(2), PGE(2), and PGI(2), in the URSA group ((2326.0 +/- 295.6) pg/ml, (2164.0 +/- 240.5) pg/ml, (238.7 +/- 26.4) pg/ml, (2337.0 +/- 263.0) pg/ml, respectively) were significantly lower than that in the control group ((3450.0 +/- 421.7) pg/ml, (3174.0 +/- 415.6) pg/ml, (323.5 +/- 43.8) pg/ml, (3623.0 +/- 460.4) pg/ml, respectively) (P < 0.05). The mRNA expression level of PPARbeta and RXRalpha (0.859 +/- 0.653, -0.172 +/- 0.752, respectively) in URSA group was significantly lower than that in the control group (1.554 +/- 1.735, 0.777 +/- 2.482, respectively) (both P< 0.05). The mRNA and protein expression levels of vascular endothelial growth factor-A (VEGF-A) in the URSA group (2.010 +/- 1.522, 0.35 +/- 0.46) was significantly lower than that in the control group (4.569 +/- 2.430, 0.750 +/- 0.350) (both P < 0.05).

CONCLUSIONSCOX-2 and the COX-2-derived PGI(2) signaling pathway possibly play an important role in successful embryo implantation, and their decreased expression may result in URSA. The decreased expression may influence the expression of VEGF-A which interferes with placental angiogenesis causing failure of embryo implantation, leading to spontaneous abortion.

Abortion, Habitual ; enzymology ; genetics ; Adult ; Blotting, Western ; Cyclooxygenase 2 ; genetics ; metabolism ; Dinoprost ; metabolism ; Dinoprostone ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Epoprostenol ; metabolism ; Female ; Humans ; Polymerase Chain Reaction ; Pregnancy ; Prostaglandin D2 ; metabolism ; Signal Transduction ; genetics ; physiology ; Vascular Endothelial Growth Factor A ; metabolism

Prostate cancer is the most common malignant tumor in male reproductive system,and androgen receptor (AR) signaling plays a critical role in the development and progression of prostate cancer.Traditional Chinese medicine shed light on the prevention and treatment of prostate cancer.Here,we focus on the relationship between AR and prostate cancer,as well as the mechanisms of traditional Chinese medicine compound,unilateral and monomer,which inhibits prostate cancer progression by regulating AR signaling.

Uveal melanoma(UM)is the most frequent and life-threatening ocular malignancy in adults.Aberrant histone methylation contributes to the abnormal transcriptome during oncogenesis.However,a comprehensive understanding of histone methylation patterns and their therapeutic potential in UM remains enigmatic.Herein,using a systematic epi-drug screening and a high-throughput transcriptome profiling of histone methylation modifiers,we observed that disruptor of telomeric silencing-1-like(DOT1L),a methyltransferase of histone H3 lysine 79(H3K79),was activated in UM,especially in the high-risk group.Concordantly,a systematic epi-drug library screening revealed that DOT1 L inhibitors exhibited salient tumor-selective inhibitory effects on UM cells,both in vitro and in vivo.Combining Cleavage Under Targets and Tagmentation(CUT&Tag),RNA sequencing(RNA-seq),and bioinformatics analysis,we identified that DOT1 L facilitated H3K79 methylation of nicotinate phosphoribosyltransferase(NAPRT)and epigenetically activated its expression.Importantly,NAPRT served as an oncogenic accel-erator by enhancing nicotinamide adenine dinucleotide(NAD+)synthesis.Therapeutically,DOT1L inhi-bition epigenetically silenced NAPRT expression through the diminishment of dimethylation of H3K79(H3K79me2)in the NAPRT promoter,thereby inhibiting the malignant behaviors of UM.Conclusively,our findings delineated an integrated picture of the histone methylation landscape in UM and unveiled a novel DOT1L/NAPRT oncogenic mechanism that bridges transcriptional addiction and metabolic reprogramming.