Objective To investigate the expression of peroxisome proliferators-activated receptor ? (PPAR?) in trophoblast and relation between PPAR? ligands and trophoblast invasion.Methods We examined the expression of PPAR? by immunohistochemistry,immunocytochemistry and real time quantitative PCR.We next examined,using the cytotrophoblast culture model,the biological role of PPAR? ligands in vitro.Results PPAR? was mainly localized in the nuclei of villous cytotrophoblast and extravillous cytotrophoblast of cell islands and cell columns.In villous tissue and cultured trophoblast from early first trimester,the level of expression of PPAR? mRNA and protein was 36.0?5.1,13.4?3.1 and 1.35?0.08,1.13?0.11;from late first trimester it was 23.3?5.5,6.1?1.3 and 1.17?0.03,0.86 ?0.05,and the expression of PPAR? was obviously decreased (P

The study was aimed to explore the role of gene JWA, a novel retinoic acid responsible and cytoskeleton associate gene, in regulating committed differentiation of HL-60 cell and the molecular mechanism in the course of differentiation and apoptosis of leukemic cells. By using FCM, the changes of CD13, CD14, CD15, CD11b and cell cycles were detected in HL-60 cells treated with ATRA (10(-6) mol/L), Ara-C (10 ng/ml) and TPA (10(-8) mol/L) respectively. The samples were determined by semi-quantitative reverse transcript-polymerase chain reaction (RT-PCR) and Western blot for the expression of JWA, Bcl-2, HSP27 and HSP70 at day 0, 2, 4, 6, 8. The results showed that HL-60 cells committedly differentiated into granulocyte-, monocyte-, macrophage-like cells. As a result, JWA was up-regulated in a time-dependent manner, while Bcl-2 was down- regulated at the same time. In ATRA and TPA group, the change of HSP70 had positive correlation with JWA, and negative correlation with Bcl-2. The expression of HSP27 was not detected. Contrast to the cells from APL patient, the expression of JWA need not be activated by ATRA in advance. In this study, we also exposed HL-60 cells in higher dose of Ara-C (20 ng/ml), and JWA expression underwent opposite trend comparing with in lower dose of Ara-C (10 ng/ml). It is concluded that JWA may play double important roles in regulating ATRA and TPA-induced differentiation and apoptosis in leukemic cells. The JWA expression had a negative correlation between induction and cytotoxic response. The difference of JWA expressions between HL-60 cell and ANLL patient cells would be involved in different leukemia pathogenesis.
Antineoplastic Agents ; pharmacology ; Blotting, Western ; Cell Differentiation ; drug effects ; Cytarabine ; pharmacology ; HL-60 Cells ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tetradecanoylphorbol Acetate ; pharmacology ; Time Factors ; Tretinoin ; pharmacology

BACKGROUNDEnhanced apoptosis of cytotrophoblasts in early pregnancy is associated with high risk of intrauterine growth retardation and preeclampsia, which are two common pregnant complications. Its etiological factors remain unclear. Cytotrophoblasts share some traits with innate immune cells and may show response to lipopolysaccharide. This study was conducted to demonstrate whether lipopolysaccharide has apoptosis-inducing effects on cytotrophoblast and the role of innate immune reaction in this process.

METHODSCytotrophoblasts were isolated from early pregnant villous tissues and cultured with serum-free medium. Subsequently, cytotrophoblasts were treated with lipopolysaccharide at the concentrations of 0 (control), 25, 50, 100 and 200 ng/ml for 24 hours. Apoptosis of cytotrophoblasts was determined by light microscopy, Hoechst 33258 DNA staining with a fluorescent microscope, transmission electron microscope and annexin V-fluorescein isothiocyanate-conjugated/propidium iodide (PI) staining with flow cytometry. Then expression of caspase-3 was detected by Western blot. Confocal immunofluorescence technique was used to detect tumor necrosis factor alpha expression in cytotrophoblasts. The levels of tumor necrosis factor alpha in the culture medium were detected by enzyme-linked immunosorbent assay.

RESULTSUnder light, fluorescence microscope and transmission electron microscope, characteristic alternations of apoptosis in cytotrophoblasts were observed after lipopolysaccharide treatment. Flow cytometry results showed that lipopolysaccharide significantly increased apoptosis indexes of cytotrophoblasts. Significant statistical differences were found in the above groups (P = 0.01). The mean relative densities of bands corresponding to caspase-3 were significantly increased in groups treated with lipopolysaccharide, as compared with the normal control (P < 0.001). Tumor necrosis factor a expression was found to increase in cytotrophoblasts by confocal immunofluorescence technique and in culture medium by enzyme-linked immunosorbent assay after lipopolysaccharide treatment. A positive correlation was found between tumor necrosis factor a expression and apoptosis indexes of cytotrophoblasts (r = 0.747, P < 0.001).

CONCLUSIONApoptosis of cytotrophoblasts could be induced by lipopolysaccharide, in which innate immune reaction is the important mechanism.

Apoptosis ; drug effects ; Blotting, Western ; Cells, Cultured ; Dose-Response Relationship, Drug ; Flow Cytometry ; Humans ; Immunity, Innate ; drug effects ; Lipopolysaccharides ; pharmacology ; Microscopy, Confocal ; Trophoblasts ; cytology ; drug effects ; Tumor Necrosis Factor-alpha ; analysis ; physiology

Objective To determine factors affecting postoperative survival of elderly patients with hepatocellular carcinoma (HCC). Methods A retrospective analysis of consecutive 54 elderly patients undergoing hepatectomy for hepatocellular carcinoma from Jan 1995 to Dec 2002 was performed. The data were analyzed by Kaplan-Meier method and Cox regression. Results Univariate analysis and Cox regression showed Child Pugh grading, vessel invasion, satellite nodule formation, Edmondson Steiner grading, intrahepatic recurrence and distant metastasis all related to postoperative overall survival or disease-free survival of the elderly with hepatocellular carcinoma (all P

OBJECTIVETo sort CD133(+) subset cells in human gastric cancer (GC) and to identify their tumor initiating cell-like properties.

METHODSThe tissues of GC and normal tissues adjacent to GC were obtained from 50 patients. Samples were stained for CD133 by immunohistochemistry. Likewise, assessments of CD133 were undertaken by Western blot. Flow cytometry was used to determine the proportion of CD133(+) cells in four GC cell lines therein the KATO-III was sorted by magnetic activated cell sorting (MACS) method. The growing characteristics and the tumorigenic ability of CD133(+) cells were evaluated in vitro and in vivo. Meanwhile, the growth of single cells in suspension culture was observed and expression of stem cell-specific marker were determined using reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe expression of CD133 was demonstrated on the cell membranes in the mucosa and submucosa of primary GC, which were higher than those in the normal gastric tissues adjacent to cancer (P<0.05). Four GC cell lines including KATO-III, SGC-7901, AGS and MKN-45 were found to contain (28 ± 2)%, (17 ± 2)%, (6 ± 2)%, and (4 ± 2)% of CD133(+) cells respectively. In addition, the purity of CD133(+) cells isolated from KATO-III by MACS was (91 ± 3)% and up to(95 ± 2)% after 1-week culture. CCK-8 detection showed that population doubling time of the CD133(+) cells was (21 ± 3)h, significantly shorter than that of the CD133(-) cells[(40 ± 8)h, P<0.05]. Notably, there was a remarkable difference of tumor formation rate between CD133(+) cells (100%), non-sorted cells (80%), and CD133(-) cells(0). The average mass and volume of tumor in group of CD133(+) cells was larger and heavier than those in non-sorted cells (P<0.05, P<0.05). Furthermore, the single cell proliferated well, formed the big sphere and semi-quantitative RT-PCR showed expression of stem cell markers such as Oct-4, Nanog, Sox-2, Musashi-1 and EGFR.

CONCLUSIONSCD133 protein expression in primary lesions is higher than those in the normal gastric tissues. CD133(+) subset cells can be isolated, purified, and amplified in human GC, and possess some properties including the ability of self-renewal, proliferation, and higher tumorigenic ability in vivo and can express some stem cell markers.

AC133 Antigen ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Middle Aged ; Neoplastic Stem Cells ; metabolism ; Peptides ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the effects of Yikunning (compound of Chinese traditional Medicine, YKN) on the apoptotic rate and expression of caspase-3 in rat ovaries during perimenopausal period.

METHODSThirty female Wistar rats during perimenopausal period were selected by unforced aging. Then the rats were divided into 3 groups randomly: YKN group, livial control group and aged control group. Ten young female rats were selected as young control group. Intragastric administrations were conducted for 4 weeks once daily continuously. The apoptotic rate in rat ovaries were detected by TUNEL. The expression of caspase-3 mRNA and protein in rat ovaries were detected by RT-PCR and Western blot, respectively.

RESULTSThe apoptotic rate in rat ovaries in YKN group was lower than that in aged control group, which showed difference between them (P < 0.01). The levels of caspase-3 mRNA and protein in rat ovaries in YKN group were lower than those in aged control group, which showed differences among them (P < 0.01).

CONCLUSIONYKN can decrease the apoptotic rate and down-regulate the expression of caspase-3 mRNA and protein in rat ovaries of during perimenopausal period. It may be one of the molecular mechanisms of YKN postponed the ovarian failure and cured perimenopausal syndrome.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Ovary ; cytology ; metabolism ; Perimenopause ; Rats ; Rats, Wistar

OBJECTIVETo evaluate the relationship between tumor necrosis factor-alpha-238, transforming growth factor beta (509 and 869) gene polymorphisms and pneumoconiosis susceptibility.

METHODSWe searched published full-text from foreign language databases including Elsevier, PubMed, Wiley Online Library, EMCC, Web of Science, chinese databases containing CNKI, VIP, Wanfang, CBM and Cochrane library to collect case-control or cohort study on gene gene polymorphisms said above with pneumoconiosis susceptibility from the year January1988 to August 2011. 28 relevant articles were selected and 20 of them met the criteria. The correlated index was extracted for aggregate analysis in RevMan 4.2.

RESULTSAmong the 20 studies, 10 articles on TNF-α238 polymorphism (including 2232 pneumoconiosis cases and 1985 control subjects), 4 articles on TGF-β509 polymorphism (including 693 pneumoconiosis cases and 663 control subjects), and 6 articles on TGF-β869 polymorphism (including 1450 pneumoconiosis cases and 1101 control subjects) were included in the current study. Meta-analysis results showed that there was a significant association between TNF-α238 polymorphism and pneumoconiosis: the population with GA and AA genotypes of TNF-α238 had higher risks to pneumoconiosis (OR = 1.53, 95%CI: 1.25 ∼ 1.88) comparing to GG genotype, and the population with A allele had higher risks to pneumoconiosis comparing to allele G (OR = 1.64, 95%CI: 1.17 ∼ 2.30). The stratified analysis showed that the people with GA and AA genotypes and A allele who were silicosis, Asian or exposed to dust had higher risks to pneumoconiosis (OR = 2.14, 95%CI: 1.20 ∼ 3.82; OR = 2.16, 95%CI: 1.20 ∼ 3.88; OR = 1.78, 95%CI: 1.01 ∼ 3.11; OR = 1.83, 95%CI: 1.04 ∼ 3.22; OR = 1.80, 95%CI: 1.21 ∼ 2.66; OR = 1.50, 95%CI: 1.23 ∼ 1.83). No significant association was found between TGF-β (509 and 869) gene polymorphisms with pneumoconiosis: In contrast to the CC genotype, the population who had CT and TT genotypes had no higher risks to pneumoconiosis (OR = 1.56, 95%CI: 0.81 ∼ 3.01; OR = 0.96, 95%CI: 0.79 ∼ 1.18); The population who had T allele had no higher risks to pneumoconiosis in contrast to the C allele (OR = 1.35, 95%CI: 0.86 ∼ 2.13; OR = 1.02, 95%CI: 0.91 ∼ 1.15).

CONCLUSIONSignificant association was found between TNFα238 gene polymorphism and pneumoconiosis; and TGF-β (509 and 869) were not.

Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Pneumoconiosis ; epidemiology ; genetics ; Polymorphism, Genetic ; Risk Factors ; Transforming Growth Factor beta1 ; genetics ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVETo observe whether there are some differences between myocardial postconditioning and remote postconditioning, and whether there is additional cardiac protection when they are used combined during myocardial ischemia/reperfusion injury in rabbits.

METHODSThirty healthy New Zealand rabbits which were randomly divided into 5 groups (n = 5): ischemic control group (CON), sham operation group (Sham), myocardial postconditioning group (MPostC), remote postconditioning group (RPostC), myocardial postconditioning plus remote postconditioning group (MPostC + RPostC). Acute myocardial infarction was induced by 45 minutes occlusion on left circumflex coronary artery (LCX) and 2 hours reperfusion in all anesthetized open-chest rabbits except the Sham, the coronary occlusion and reperfusion were determined by changes of ECG and cardiac color. Skeletal muscle ischemia model was induced by extrinsic iliac arteries occlusion and reperfusion with artery clamps. The condition that the extrinsic iliac arteries were occluded or reperfused could be tested by according to the distal arterial pulse. Plasma creatine kinase (CPK) activity and lactate dehydrogenase (LDH) activity were measured at baseline, the end of ischemia, after 1 hour and 2 hours of reperfusion respectively. The extent of myocardial infarction was assessed by triphenyltetrazolium (TTC) staining and measured by area ratio of AN/AAR.

RESULTSCompared with the Con, myocardial infarct size was significantly reduced in MPostC and RpostC group (P < 0.05). But there was no significant difference between MPostC and RPostC group. Combined MPostC and RPostC markedly enhanced myocardial protection (P < 0.05). The trend of CPK and LDH release was similar to the trend of myocardial infarct size.

CONCLUSIONBoth MPostC and RPostC induced cardiac protection. There was no significant difference between MPostC and RPostC. Combined MPostC and RPostC induced markedly additive effect on myocardial protection.

Animals ; Disease Models, Animal ; Ischemic Postconditioning ; Muscle, Skeletal ; blood supply ; Myocardial Reperfusion Injury ; prevention & control ; Myocardium ; metabolism ; Rabbits

OBJECTIVETo study the feasibility and validity of multi-modal serial therapy for primary liver cancer in senile patients.

METHODS153 senile primary liver cancer patients (>or= 60 years) were given multi-modal serial therapy from June 1993 to December 2000. Hepatectomy was performed in 37, deep cryosurgery in 32 and non-operative therapy in 84 (intervention as chief therapy in 81, combined local and intervention therapy in 3). The multi-course intervention therapy was given postoperatively in hepatectomy and cryosurgery groups, while bioimmunotherapy and traditional Chinese medicine were used in all groups.

RESULTSThe 1-, 3- and 5-year survival rates in the hepatectomy group were 78.4%, 46.4% and 35.7%, without operative mortality. The 1- and 3- and 5-year year survival rates in the cryosurgery group were 64.5%, 40.9% and 25.0% with mortality of 3.1%. Among patients with non-operative therapy, the 1- and 3- and 5-year year survival rates in intervention group were 47.5%, 23.5% and 4.3%. The operative mortality was 1.2%. The 3 patients who received combined local and intervention therapy have survived for 2.5, 3.8 and 7.1 years.

CONCLUSIONMulti-modal serial therapy with surgical treatment as the chief means, being precise in the effect and good in safety, is feasible and valid for primary liver cancer in senile patients.

Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Carcinoma, Hepatocellular ; pathology ; therapy ; Chemoembolization, Therapeutic ; Combined Modality Therapy ; Cryosurgery ; Feasibility Studies ; Female ; Hepatectomy ; Humans ; Infusions, Intra-Arterial ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Staging ; Survival Rate