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.

OBJECTIVETo construct primary cultured granulosa cells model of Zi Gooses tansfected by alpha-enolase (ENO1) overexpression adenovirus vector, and to detect the effect of ENO1 overexpression of granulose cells on progesterone secretion.

METHODSGranulosa cells were infected with Ad-CMV-ENO1 in gradient multiplicity of infection(MOI) levels:100, 250, 350 and 400 pfu/cell. Twenty four hours and 48 h after infection, green fluorescent protein (GDP) was respectively detected by fluorescence inverted microscopy. The effect of ENO1 overexpression of granulose cells on progesterone secretion was detected by the step double antibody sandwich enzyme-linked immunosorbent assay (ELISA).

RESULTSThe optimal infection rate (100%) was achieved when MOI was 800 pfu/cell,48h after infection. Real time RT-PCR and Western blot showed that the level of mRNA and protein expression ENO1 were increased significantly after infection (P < 0.01); The granulosa cells progesterone secretion of Ad-CMV-ENO1 group increased signigicantly (P < 0.01).

CONCLUSIONENO1 overexpression could make the primary culture follicle granulosa cells in vitro improve progesterone secretion.

Adenoviridae ; Biomarkers, Tumor ; genetics ; Cell Line ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; Female ; Genetic Vectors ; Granulosa Cells ; metabolism ; Green Fluorescent Proteins ; genetics ; Humans ; Ovarian Follicle ; cytology ; Phosphopyruvate Hydratase ; genetics ; Progesterone ; secretion ; RNA, Messenger ; genetics ; Transfection ; Tumor Suppressor Proteins ; genetics

Objective To establish the ischemic precondition ([PC) model of PC12 cell line in vitro, and to explore the effect of nitric oxide (NO) on the IPC cerebral protection. Method PC12 cells were cultured and used for producing the model of ischemie precondition by the way of oxygen-glucose deprivation. Twenty dishes of cells were randomly divided into four groups (5 dishes for each group): control group, ischemic precondition group (IPC),non-ischemic precondition group (NIPC) and L-NAME treatment group (L-NAME). In control group, the cells were in-cubated with low glucose (<1 g/L) and2% FBS medium in normal oxygen; in IPC group, the cells were administrated with oxygen-glucose deprivation (OGD) for 6 hours, and then subjected with reperfuaion before OGD 15 hours; in NIPC group, the cells were treated the same as control group for 6 hours, and then subjected with reperfusion before OGD 15 hours; in L-NAME group, the cells received L-NAME (1 mmol/L) and cocultured for 30 minutes before OGD 6 hours, and then received the same treatment as the IPC group. To test whether the model was established, metabolic rate of MIT, LDH release were measured and the apoptosis rate was detected by flow cytometry following oxygen-glucose deprivation 15 hours. The activity of nitric oxide synthases (NOS) was as-sessed by biochemical assay. One-way ANOVA and LSD multiple comparison test were used to analyze differences among different groups, and P<0.05 was considered different. Results Compared with NIPC group, the metabolic rate of MTT increased (94.9%±35.1%, P<0.05), while LDH release and the cell apoptotic rate decreased significantly in IPC group (279.1%±28.1%, P<0.03). Compared with control group(100.0%± 13.5%),the activities of NOS increased both in NIPC and IPC groups (390.0%±14.6%, P<0.01;126.3% ±10.6%, P<0.01). Moreover, the apoptosis rates in each group (control group, IPC group, NIPC group and L-NAME group) were 5.90, 8.73, 38.62 and 11.73%,respectively. Conclusions IPC reduces the death and apoptosis rate of PC12 cell after oxygen-glucose deprivation injury. NO might be involved, but it is not the only factor.

As a neuropeptide, neurotensin (NTS) is widely expressed in central and peripheral nervous system, which is mainly mediated byneurotensin receptor1 (NTSR1) to activate the related downstream signaling pathways. After summarized the function and mechanism of NTS/NTSR1 in various malignant tumors, we found that NTS/NTSR1 played essential roles during tumor initiation and development. NTS/NTSR1 regulates tumor initiation, proliferation, apoptosis, metastasis and differentiation mainly through three pathways, including IP3/Ca2+ /PKC/MAPKs pathway, MMPs/EGFR/MAPKs (PI3K/Akt) pathway, or Rho-GTPsaes and non-receptor tyrosine kinase pathway. Besides, NTS/NTSR1 is also regulated by some upstream pathways and some traditional Chinese medicine preparations and traditional Chinese medicine therapies. In this article, we summarized the function of NTS/NTSR1 and its mechanisms, and discussed the prospective in its application to clinical diagnosis and drugs targeting.
Animals ; Humans ; Medicine, Chinese Traditional ; Neoplasms ; etiology ; Neurotensin ; chemistry ; physiology ; Receptor, Epidermal Growth Factor ; physiology ; Receptors, Neurotensin ; chemistry ; physiology ; Signal Transduction ; physiology ; rhoA GTP-Binding Protein ; physiology

Objective:To investigate the feasibility of targeted imaging and therapy of prostate cancer using nanocomposite probes composed of fluorescent magnetic nanoparticles(FMCNPs) and single chain Fv(ScFv) antibody specific for gama-seminoprotein.Methods:The nanocomposite probes(FMCNPs-ScFv) were prepared by conjugating fluorescent magnetic nanoparticles with singlegama-chain Fv antibody specific gama-seminoprotein,and were characterized by high resolution transmission electron microscopy,fluorescent spectrum and magnetic spectrum.Nanocomposite probes were incubated with prostate cancer LNCaP cells,and the targeting results of nanocomposite probes were observed by fluorescent microscopy.The cytotoxicity effect of the nanocomposite probes was measured by MTT.Nude mice models of prostate cancer were established and identified by immunohistochemistry method.The nanocomposite probes were injected into nude mice via tail vein.The distribution of nanocomposite probes in the nude mice was observed by Micro-animal imaging system,targeted imaging of the prostate cancer was observed by MR instrument.The nude mice with prostate cancer were irradiated with 100 W magnetic field for 30 min,and the changes of tumor sizes were observed.Results:The FMCNPs-ScFv nanocomposite probes were successfully prepared.Nanocomposite probes entered into the cytoplasm of cancer cells and exhibited low cytotoxicity effect.Nude mice model with prostate cancer were successfully fabricated;the nanocomposite probes distributed quickly in the main organs of mice,and gradually concentrated on the tumor tissues within 24 h.MR images showed that the tumor images were gradually enhanced from 6 h to 24 h after injection of the nanocomposite probe.Four days after magnetic irradiation,the tumors in the nude mice grew slower compared with the control nude mice(P

OBJECTIVETo evaluate the levels of bone morphogenetic protein-15 (BMP-15) in human follicular fluid (FF) and its association with response to ovarian stimulation.

METHODSWestern blotting was performed to determine the levels of BMP-15 in FF obtained from follicle aspirates in 70 patients undergoing IVF treatment. According to the response to ovarian stimulation the patients were divided into poor responder group and normal responder group.

RESULTBMP-15 levels in FF of poor responders were significantly higher than those in normal responders (1.01 +/- 0.34 vs 0.77 +/- 0.24, P<0.01).

CONCLUSIONIncreased levels of BMP-15 in FF may be associated with poor response to ovarian stimulation.

Adult ; Blotting, Western ; Bone Morphogenetic Protein 15 ; Female ; Follicle Stimulating Hormone ; administration & dosage ; Follicular Fluid ; drug effects ; metabolism ; Gonadotropin-Releasing Hormone ; administration & dosage ; analogs & derivatives ; Growth Differentiation Factor 9 ; Humans ; Infertility, Female ; metabolism ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Ovary ; drug effects ; metabolism ; Ovulation Induction

Animals ; Cell Hypoxia ; Cell Survival ; drug effects ; Cells, Cultured ; Female ; Hippocampus ; cytology ; drug effects ; Male ; Neurons ; cytology ; drug effects ; Neuroprotective Agents ; pharmacology ; Peptides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo classify the Chinese isolates of Coltiviruses.

METHODSThree sets of primers were selected among them two were specific to the 9th and 12th segments of subgroup B2, and one was for the 12th segment of subgroup B1-All the Chinese isolates of Coltivirus selected in the experiment were classified according to the lengths of different amplicons of the reverse transcriptase-polymerase Chain reaction (RT-PCR). The homogenicity of the nucleic acids of the isolates BJ95-75 and YN-6 was also compared with other Coltivirus strains belonging to subgroup B2.

RESULTSWith the primers 12-854-S/12-B2-R, which were specific to the 12th segment of Coltivirus subgroup B2-850 bp amplicons were obtained from Beijing isolate BJ95-75 and all the Yunnan isolates such as YN-6, -67-1, -68-1, -69, -70-1, -70-2, -90, -92-2, -93 of Coltivirus 492 bp DNA fragments were also amplified from all of them with the segment 9th specific primers 9-JKT-S/9-JKT-R. However no positive results were obtained from Northeast isolates NE97-12, NE97-31 and control viruses YN-99(Orbivirus),YN-151-1(JEV) with the same two sets of primers. With 12-B1-S/12-B1R primers specific to the 12th segment of subgroup B1, no amplicons of right length were obtained from any of the Chinese isolates of Coltivirus and the control viruses. When compared the nucleic acid sequences of BJ95-75 and YN-6 with other Coltivirus strains such as Bannavirus, JKT6423, JKT6969, JKT7043, the amplicons from segment 12th of these two strains had more than 89.4% homology with the other strains, especially to the earlier Chinese isolate Bannavirus, the homolog was more then 98.9%. Nearly 96.5% and 99.2% of the nucleic acids of the amplicons from segment 9th of the two strains were being homologous to Bannavirus and about 84.0% to JKT6423, which had been classified into type B2a. But the maximal homogenicity was about 53% when compared with the other two coltivirus strains. JKT6969 and JKT7043 which had been classified into type B2b.

CONCLUSIONGenotyping the recent Chinese isolates of coltivirus for the first time in our country. Most of the Chinese isolates belong to subgroup B2, more exactly type B2a. The Northeast isolates NE97-12 and NE97-31 were not correctly grouped with the available primers.

Animals ; Base Sequence ; China ; Coltivirus ; classification ; genetics ; isolation & purification ; Culicidae ; virology ; Genotype ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Nucleic Acid

OBJECTIVETo investigate whether or not adaptive response of hPARP-1 protein normal and deficient cells is induced by low dose of hydroquinone (HQ), and to analyze the relationship between the adaptive response and micronuclei formation, and cell cycle alteration in human embryo lung fibroblasts (HLF), so as to elucidate the mechanism of adaptive response.

METHODSHLF, HLFC and HLFP cells pretreated with low concentration were retreated by high concentration of HQ. Cellular viability, the rate of micronuclei and abnormal nuclei, cell cycle and DNA strand break were determined.

RESULTSThe tolerance to 80.0 micromol/L concentration of HQ was enhanced when HLF, HLFC and HLFP cells were pretreated with HQ from 0.001 - 0.050 micromol/L. There were varying degrees of micronuclei and abnormal nuclei in three cells pretreated with low concentration of HQ and then retreated with high concentration of HQ; the cell numbers of G1, G2, S phase in cell cycle were obviously different. When compared with only high attack dose, the micronuclei rate and abnormal nuclei rate of HLF, HLFC and HLFP decreased by pretreatment with HQ at high concentration (P < 0.05), meanwhile increased by pretreatment with HQ at low concentration (P < 0.05). HLF, HLFC and HLFP showed blockage in G2 phase when pretreated with HQ at 0 approximately 0.05 micromol/L, but HLFP showed blockage in G1 phase, and in S phase at 1.0 and 2.0 micromol/L.

CONCLUSIONThe level of adaptive response of hPARP-1 protein deficient cells was lower than normal cell, suggesting that hPARP-1 protein may play an important role in the adaptive response of cells, which may be related with the regulation of cell cycle.

Cell Cycle ; Cell Nucleus ; Cell Survival ; Cells, Cultured ; Dose-Response Relationship, Drug ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Hydroquinones ; toxicity ; Lung ; cytology ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism

To investigate the antitumor activities of the immunoconjugates composed of anti-type IV collagenase monoclonal antibody Fab' fragment and lidamycin (LDM) prepared with different linkers. The immunoconjugates were prepared by linking Fab' to lysine-69 of LDM apoprotein by SPDP, LCSPDP, SMBS or SSMPB as the intermediate drug linkers. Immunoreactivities of the conjugates were determined by ELISA. The cytotoxicities of the conjugates were examined by clonogenic assay. In vivo antitumor effects of the conjugates were evaluated in nude mice bearing subcutaneously implanted HT-1080 tumor. ELISA assay showed that the conjugates retained part of the immunoreactivity of 3G11 against the antigen. The cytotoxicities of the Fab'-SMBS-LDM and Fab'-SSMPB-LDM to HT-1080 cells were significantly potent, compared with Fab'-SPDP-LDM, Fab'-LCSPDP-LDM and free LDM. In animal models at the same condition, free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM inhibited the growth of HT-1080 tumor by 70.9%, 74.8% and 72.3%, while Fab'-SMBS-LDM and Fab'-SSMPB-LDM reached 78.0% and 87.7%, respectively. The median survival time of the mice treated with free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM were prolonged by 71.9%, 82.2% and 107.5%, respectively, compared with that of untreated group. Whereas, the median survival time of Fab'-SMBS-LDM and Fab'-SSMPB-LDM were prolonged by 145.2% and 165.8%, respectively, indicating that Fab'-SSMPB-LDM was more effective than Fab'-SMBS-LDM in tumor suppression and life span prolongation. Fab'-SSMPB-LDM has more marked selective antitumor efficacy and lower toxicity, and might be a novel candidate for cancer therapy.
Aminoglycosides ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antibodies, Monoclonal ; immunology ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Collagenases ; immunology ; Enediynes ; pharmacology ; Fibrosarcoma ; pathology ; Humans ; Immunoconjugates ; pharmacology ; Immunoglobulin Fab Fragments ; immunology ; Matrix Metalloproteinase Inhibitors ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Tumor Burden ; drug effects