Objective:To investigate the expressions of peripheral lymph node addressin(PNAd)andGlcNAc-6-sulfotransferase(GlcNAc6ST)in endometrium and their impacts on implantation.Methods:PNAd expression in endometrium was examined by immunohistochemistry and Western Blot from 75women(12 from healthy women,in proliferative phase;63 from sterile women,of whom,27 were inearly-secretory and 36 in mid-secretory phase).GlcNAc6ST mRNA was examined by real-time PCR in41 sterile women.The 63 sterile women had underwent ⅣF-ET and were consequently divided into clini-cal pregnant(29 cases)and nonpregnant(34 cases)groups.Results:(1)PNAd localized to the mem-brane and cytoplasm of luminal and glandular epithelia.Staining was patchy and much less intense duringthe proliferative phase than during the secretory phase.In Western Blot of PNAd,four bands appeared,which were Sgp200,CD34,MAdCAM-1,GlyCAM-1 respectively,and each was positively correlatedwith the others significantly.The former three molecular levels were significantly higher during the secre-tory phase as compared with the proliferative phase.Message RNA of GlcNAe6ST was positive in all ca-ses and showed no correlation with any component of PNAd.(2)The expressions of CD34 and GlyCAM-1,but not Sgp200 and MAdCAM-1,were significantly higher in pregnant women than in nonpregnantones.However,the GlcNAc6ST mRNA level did not differ between groups.(3)No significant differ-ence was found in female age,methods of fertilization,thickness of endometrium on day hCG,cumulativeembryo score(CES)and mean score of transferred embryo(MSTE)between the groups.Conclusion:PNAd expression in the human endometrium fluctuates with the menstrual cycle.Elevated CD34 and Gly-CAM-1 during the secretory phase might be stimulative factors for embryo implantation.Defect in PNAdexpression may account for a portion of unexplained infertility.

Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Female ; Fetal Blood ; Humans ; Infant, Newborn ; Lipopolysaccharides ; Lymphocytes ; metabolism ; Male ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; genetics ; metabolism ; Time Factors ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To investigate the changes of hypoxia-inducible factor(HIF)-1α and hexokinase-Ⅱ(HK-Ⅱ)expression in human esophageal squamous cell carcinoma and its effect in glycolysis.Methods TE13 cells and Eca109 cells were cultured under hypoxic condition(1 ％O2)for different hypoxic time(6,12,24 and 48 hours).Cells cultured under normal oxygen condition(20％O2)were set as control.The changes of HIF-1α and HK-Ⅱ expressions at protein level were detected by Western blot.HIF-1α genes were specifically silenced with RNA interference technology(RNAi),and then the changes of HIF-1α and HK-Ⅱ expression were determined by realtime PCR and Western blot.Under normal oxygen and hypoxic condition,the changes of lactic acid concentration in cell culture medium were detected by spectrophotometric method.Results Under hypoxic condition,the expression of HIF-1α and HK-Ⅱ gradually increased as hypoxic time extended(P＜0.05),reached a peak at 12h and then gradually decreased as time extended.Compared with that under normal oxygen condition,the expression of HK-Ⅱ in TE13 cells and Eca109 cells significantly increased under hypoxic condition(P＜0.05),which was more significant after 12 hours hypoxia.The result of realtime PCR indicated that under normal oxygen condition the expression of HIF-1α at RNA level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference(P＜0.05).The expression of HK-Ⅱ at RNA level was consistent with the result of HIF-1α.Under normal and hypoxia condition,the expression of HK-Ⅱ at protein level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).The lactic acid secretion of TE13 cells and Eca109 cells under hypoxia condition(14.707 ± 3.594 and 15.062 ±3.901)was higher than that under normal oxygen condition(6.070±1.839 and 6.891±1.592,P＜0.05).The lactic acid secretion of TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).Conclusion The expressions of HIF-1α and HK-Ⅱ in human esophageal squamous cell carcinoma significantly increased under hypoxia conditions.The expression of HK-Ⅱ is closely correlated with lactic acid concentration and HIF-1α expression.HIF-1α may affect cell glycolysis through HK-Ⅱ.

OBJECTIVETo compare the difference in clinical efficacy on children attention deficit hyperactivity disorder (ADHD) between the combined therapy of scalp acupuncture and EGG biofeedback and the simple EEG biofeedback therapy so as to search the better therapeutic method for ADHD.

METHODSOne hundred patients were randomized into an observation group and a control group, 50 cases in each one. In the control group, the simple EEG biofeedback therapy was adopted. In the observation group, on the basis of biofeedback therapy, scalp acupuncture was added and applied to Dingzhongxian (MS 5), Dingpangyixian (MS 8), Baihui (GV 20), Sishencong (EX-HN 1), etc. The ten treatments made one session. After four sessions of treatment, FIQ value in Wechsler intelligence scale, CIH score in Conners children behavior questionnaire, the ratio of 0 wave and p wave in EEG, FRCQ and FAQ in the integrated visual and auditory continuous performance test (IVA-CPT) and clinical comprehensive efficacy were observed before and after treatment in the two groups separately.

RESULTSThree cases were dropped out in the observation group and 2 cases were out in the control group. In the two groups, FIQ, FRCQ and FAQ were all increased after treatment (P < 0.01, P < 0.05); the increases in the observation group were much more significant than those in the control group after treatment (all P < 0.05). In the two groups, CIH score and the ratio of 0 wave and p wave were all reduced after treatment (P < 0.01, P < 0.05); the reduction in the observation group were much more apparent as compared with those in the control group (both P< 0.05). The total effective rate was 91.5% (43/47) in the observation group and better than 83. 3% (40/48, P < 0.01) in the control group.

CONCLUSIONThe combined therapy of scalp acupuncture and EEG biofeedback achieves the superior efficacy on children ADHD as compared with the simple biofeedback therapy. This combined therapy rapidly relieves the essential symptoms of ADHD and improves EEG waveform in children patients. Importantly, this therapy obtains and consolidates the significant efficacy.

Acupuncture Therapy ; Adolescent ; Attention Deficit Disorder with Hyperactivity ; psychology ; therapy ; Biofeedback, Psychology ; Child ; Combined Modality Therapy ; Electroencephalography ; Female ; Humans ; Male ; Scalp

Objective To observe the quantity change of autophagosomes in podocytes and expressions of autophagy-related gene Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) in different pathological stages of idiopathic membranous nephropathy (IMN),and to explore how autophagy is related to podocyte injury,the occurrence of proteinuria and the disease progression in IMN.Methods Clinical data of 26 patients who were diagnosed as IMN (14 IMN stage 1 and 12 IMN stage 2) admitted to Zhejiang Provincial people's Hospital from January 2013 to December 2014 were retrospectively analyzed.Normal renal tissue from 15 cases of kidney neoplasms with nephrectomy was collected as control.The changes of kidney tissue pathology were detected after PAS and PASM staining by light microscope.The autophagosomes of podocyte were detected by transmission electron microscopy.Expressions of Beclin-1 and LC3 protein were detected by immunohistochemistry.Expressions of LC3 and synaptopodin were detected by immunofluorescence.The correlation of autophagosomes and clinical pathologic factors in IMN patiens was analyzed.Results There were fewer autophagosomes of podocytes and lower expression of Beclin-1 and LC3 protein in IMN group than those in control group (P=0.034,P=0.011,P=0.013,respectively).Moreover,these effects were more obvious with the development of IMN.Compared with those in control group,autophagosomes,Beclin-1 and LC3 protien were reduced in IMN stage 2 group (P=0.009,P=0.030,P=0.015);the number of autophagosomes and the expressions of LC3 and Beclin-1 were decreased in IMN stage 1 group as well,however statistically insignificant (P=0.352,P=0.087,P=0.128);Comparisons between IMN stage 2 patients and IMN stage 1 patients shown significant difference in the number of autophagosomes (P=0.030),but no significant difference in expressions of Beclin-1 and LC3 (P=0.355,P=0.181).Autophagosomes number was not correlated with serum creatinine,serum urea nitrogen,24-hour urinary protein and eGFR (all P ＞ 0.05).The expressions of synaptopodin and LC3 protein were lower in IMN group than those in control group.Conclusion Autophagy may contribute to podocyte injury and the production of protein urine in IMN,and may be closely related to the progression of disease.

AIM: To observe the effect of protein kinase C-?(PKC?)antisense oligonucleotide on cell growth, cell cycle and the expression of cyclin E in human poor-differentiated nasopharyngeal carcinoma(NPC) cell line CNE-2Z. METHODS: Antisense PKC? was transfected by cationic liposome(LP) in CNE-2Z cells to analyze the cell growth and cell cycle by MTT colorimetric assay and flow cytometry, respectively. Moreover, the expression of cyclin E was determined by immunocellularchemistry and scanning the result of dot-blotting. RESULTS: ①With the concentration of antisense PKC? increasing, the relative cell growth index was decreased gradually( P

Objective To investigate the correlation between the expression of HIF-1a (hypoxia inducible factor 1,HIF-1a) in perihematomal brain issue and formation of brain edema in patients with hypertensive cerebral hemorrhage.Method Perihematomal brain issue was collected in the course of hematoma elimination in 32 patients with hypertemive intraeerebral hemorrhage.Expressions of HIF-1a and vascular endothelial growth factor (VEGF) were observed by immunohistochemistry.The volume of perihematomal brain edema on computed tomographie scan was determined by computed tomographic scan before surgery.The results of staining and the volume of perihematomal brain edema were analyzed in double blind fashion.Results HIF-1a protein immunohistochemical staining positive cells (2.8?0.8/HP) were identified dispersedly from 4 hours after acute hemorrhagic stroke in perihematomal brain issue,and reached the peak at 24～48 hours (12.5?3.9/HP).High expression of HIF-1a progressed at 48～72 hours (12.2?1.8/HP) after acute hemorrhagic stroke.There was a positive correlation between the expression of HIF-1a and VEGF (r=0.76,t=6.37,P

OBJECTIVETo investigate the expression profile of interleuki-1β (IL-1β) in rat myocardium at different time points during hypoxia/reoxygenation(H/R)transition.

METHODSThe isolated Langendorff perfused rat heart model was established.Forty SD rats were randomly divided into sham group (A group) and hypoxia/reoxygenation group (H/R group). The H/R group rats were subdivided into H/R 0.5 h group(B group), H/R 1 h group(C group), H/R 2 h group(D group)according to reoxygenation time. The left ventricular development pressure(LVDP), maximal rates of increase/decrease of the left ventricular pressure(±dp/dtmax) were continuously recorded. The concentration of interleukin-1β(IL-lβ) and creatine kinase-MB (CK-MB) in myocardium was measured by ELISA. The mRNA expression of IL-lβ in myocardium was determined by RT-PCR. Microstructure of myocardium was observed under light microscopy.

RESULTSThe value of LVDP and ±dp/dtmax in hypoxia/reoxygenation group rat were significantly lower than that in sham group(P < 0.05). The expression of IL-lβ and CK-MB at protein level and the expression of IL-1β at mRNA level in hypoxia /reoxygenation group were higher than that in sham group(P < 0. 05). There were significant differences of the above parameters among H/R 0.5 h, 1 h, 2 h group(P <0.05). The concentration of IL-1β and CK-MB, the mRNA expression of IL-1β were higher in H/R 2 h group than that of other groups(P < 0.05).

CONCLUSIONThe high expression of IL-Iβ in myocardium after myocardial hypoxia /reoxygenation in rats might lead to. ischemia/reperfusion injury.

Animals ; Creatine Kinase, MB Form ; metabolism ; Disease Models, Animal ; Hypoxia ; metabolism ; pathology ; Interleukin-1beta ; metabolism ; Myocardial Ischemia ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

This study was to investigate the chemical constituents of the aerial part of Zygophyllumfabago, by phytochemical methods. The compounds were isolated by silica gel and Sephadex LH-20 column chromatographies from the EtOAc extract. Their structures were characterized by various spectroscopic data (1H-NMR, 13C-NMR, MS) and comparison with the literature. As a result, thirteen compounds were isolated and their structures were identified as 1-hydroxyhinesol(1), hinesol(2), atractylenolactam(3), beta-eudesmol (4), 5alpha-hydroperoxy-beta-eudesmol(5), 12-hydroxy-valenc-1(10)-en-2-one(6), pubinernoid A(7), (6S,7E)-6-hydroxy-4,7-megastigmadien-3,9-dione(8), 3-hydroxy-5alpha, 6alpha-epoxy-beta-ionone (9), (3S,5R, 6S, 7E)-3, 5, 6-trihydroxy-7-megastigmen-9-one(10), (6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one(11), (S)-3-hydroxy-beta-ionone(12), and blumenol A(13). Compounds 1-7 were sesquiterpenoids and 8-13 were megastigmane type norsesquiterpenoids. All the compounds were obtained from Z. fabago for the first time, and compound 1 was a new natural product.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Terpenes ; chemistry ; isolation & purification ; Zygophyllum ; chemistry

OBJECTIVETo investigate the role of p38 MAPK on ischemic postconditioning (IPO) attenuating pneumocyte apoptosis after lung ischemia/reperfusion injury (LIRI).

METHODSForty adult male SD rats were randomly divided into 5 groups based upon the intervention (n = 8): control group (C), LIR group (I/R), LIR + IPO group (IPO), IPO + solution control group (D), IPO + SB203580 group (SB). Left lung tissue was isolated after the 2 hours of reperfusion, the ratio of wet lung weight to dry lung weight (W/D), and total lung water content (TLW) were measured. The histological structure of the left lung was observed under light and electron transmission microscopes, and scored by alveolar damage index of quantitative assessment (IQA). Apoptosis index (AI) of lung tissue was determined by terminal deoxynuleotidyl transferase mediated dUTP nick end and labeling (TUNEL) method. The mRNA expression and protein levels of and Bax were measured by RT-PCR and quantitative immunohistochemistry (IHC).

RESULTSCompared with C group, W/D, TLW, IQA, AI and the expression of Bax of I/R were significantly increased, the expression of Bcl-2 and Bcl-2/Bax were significantly decreased (P < 0.05, P < 0.01), and was obviously morphological abnormality in lung tissue. Compared with I/R group, all the indexes of IPO except for the expression of Bcl-2 and Bcl-2/ Bax were obviously reduced, the expression of Bcl-2 and Bcl-2/Bax were increased (P < 0.05, P < 0.01). All the indexes between D and IPO were little or not significant( P > 0.05). The expression of Bcl-2 and Bcl-2/Bax of SB were significantly increased and other indexes were reduced than those of IPO (P < 0.05, P < 0.01).

CONCLUSIONIPO may attenuate pneumocyte apoptosis in LIRI by inactivation of p38 MAPK, up-regulating expression of Bcl-2/Bax ratio.

Alveolar Epithelial Cells ; cytology ; Animals ; Apoptosis ; Disease Models, Animal ; Ischemic Postconditioning ; Lung ; blood supply ; enzymology ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology ; pathology ; prevention & control ; bcl-2-Associated X Protein ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism