Objective To investigate the expression of adiponectin in placenta and its correlation with preeclampsia.Methods Placental tissues were collected from normal term pregnancies(normal pregnancy group,n=20),mild preeclampsia(mild preeclampsia group,n=12)and severe preeclampsia (severe preeclampsia group,n=22).The expression of adiponectin protein and the intensity of its mRNA in placenta were detected using immunohistochemistry and RT-PCR,respectively.Integral optical density (IOD)which represents the expression level of adiponectin protein,and the ratio of adiponectin cDNA PCR products to β-actin cDNA PCR products which represents the intensity of transcription of adiponectin mRNA in placenta were analyzed.Results (1)The expression of adiponectin protein was observed in cytoplasm of placental cytotrophoblasts and syncytiotrophoblasts among three groups.There was no significant difference in adiponectin protein expression between maternal side and fetal side of placenta in three groups(all P＞0.05);(2)The expression of adiponectin protein in placenta in severe preeclampsia group(30 984 ±14 604)was significantly lower than that of mild preeclampsia group(58 360±8910,P＜0.01)and of normal pregnancy group(53 246±17 554,P＜0.01).There was also no significant difference in the expression of adiponeclln protein in placenta between term delivery and preterm delivery in severe preeclampsia group(38 890±20 386 vs 29 319±8997,P＞0.05),however,the expression of adiponectin protein in placenta in term delivery of severe preeclampsia group was significantly lower than that ofterm delivery of normlal pregnancy group(38 890±20 386 vs 53 246±17 554,P＜0.05);(3)The expression of adiponectin mRNA was detected in placental tissues among three groups also.The intensity of transcription of adiponectin in placenta in severe preeclampsia group(1.0±0.2)was markedly lower than that of mild preeclampsia group(2.9±0.8,P＜0.05)and normal pregnancy group(3.3±1.1,P=0.000).Conclusion The expression of adiponectin decreases in placenta tissues of severe preeclampsia,indicating that the abnormal expression of adiponectin may be involved in the pathogenesis of preeclampsia.

OBJECTIVETo investigate the effects on chemotherapeutic sensitization of the PI3K inhibitor LY294002 in diffuse large B cell Lymphoma (DLBCL) cell lines ly1, ly8, ly10.

METHODSThe three cell lines were treated with LY294002, or doxorubicin alone or combined or sequentially respectively. Western blotting was used to detect the level of phospho-AKT after the treatment. Flow cytometry combined with annexin V-FITC assay and Brdu incorporation assay were used to analyze the alterations of cell cycle, proliferation, and apoptosis, respectively.

RESULTSLY294002 decreased the level of phospha-AKT efficiently in the three DLBCL cell lines. The ratio of S phase cells was significantly decreased (P < 0.05). Sequential use of LY294002 and doxorubicin increased the ratio of apoptosis and there was significant difference between the sequential group and the other four groups (P < 0.05) at 24, 48, 72(ly1), 48, 72 (ly8) or 24 h (ly10).

CONCLUSIONLY294002 can sensitize doxorubicin-induced apoptosis and may be a potential molecular therapeutic agent targeted at AKT signaling pathway in DLBCL.

Apoptosis ; drug effects ; Cell Line, Tumor ; Humans ; Lymphoma, Large B-Cell, Diffuse ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Proto-Oncogene Proteins c-akt ; metabolism

OBJECTIVETo observe the status of AKT and phospho-AKT (pAKT) in three diffuse large B cell lymphoma (DLBCL) cell lines, and to investigate the effects of AKT activation on biologic behavior of DLBCL cells.

METHODSThree DLBCL cell lines, ly1, ly8 and ly10 were maintained in 10% FBS or serum free culture medium. The expression of AKT and status of pAKT were detected by Western blotting. LY294002, an inhibitor of PI3K, was used to suppress the level of pAKT. Flow cytometry combined with PI staining, AnnexinV-FITC assay and Brdu incorporation assay were used to analyze the parameters of the cell cycle, apoptosis and proliferation respectively.

RESULTSThere was constitutive activation of AKT in three DLBCL cell lines and the levels of pAKT were altered in the different environments. In 10% FBS culture medium, pAKT was higher than that in serum free culture medium in ly8 and ly10, however, pAKT in ly1 maintained in serum free culture medium was mildly higher than that in 10% FBS culture medium. When the cell lines ly1, ly8, ly10 were maintained in 10% FBS culture medium, the inhibitor LY294002 suppressed the level of pAKT efficiently in three DLBCL cell lines. The percentage of cells at S phase and the proliferation index were significantly decreased (P < 0.05) without an increase of apoptosis (P > 0.05).

CONCLUSIONSActivation of AKT may play an important role in the development of DLBCL. It is closely related to the control of cell cycle and proliferation, but is not associated with apoptosis. LY294002 can inhibit cell growth by decreasing the levels of pAKT in DLBCL cell lines.

Apoptosis ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromones ; pharmacology ; Culture Media, Serum-Free ; Enzyme Activation ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism

OBJECTIVE: To explore the change rules of peak area ratio of STR loci to Amelogenin (AMEL) locus (STR/AMEL), a sex-determining gene in DNA degradation, and to evaluate the application of STR/AMEL value in the estimation of DNA degradation degree. METHODS: DNA was extracted from iliopsoas, and the variations of STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) were analyzed after the artificial degradation was made by DNase I, and the changes of these three ratios of the iliopsoas naturally degraded in an outdoor environment were also analyzed. The regression curves were analyzed using the periods of DNA degradation and outside the body as the independent variable (x) and the STR/AMEL value as the dependent variable (y) and three curve equations under two conditions were established. RESULTS: Both under the conditions of artificial and natural degradation, STR/AMEL value had a negative relationship with the degradation time. The relationship between STR/AMEL and degradation time can be well simulated by the cubic function. R2 was over 0.99 under controlled degradation condition and over 0.86 under natural degradation condition. CONCLUSION The STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) is negatively related with the DNA degradation degree, which follows mathematical regression models strictly, and it might be applied to evaluate the DNA degradation degree.
Amelogenin/genetics* ; DNA Damage/genetics* ; DNA Primers ; Humans ; Microsatellite Repeats ; Regression Analysis ; Time Factors

OBJECTIVETo explore the effect of nitric oxide (NO) on human sperm capacitation and acrosome reaction (AR).

METHODSDifferent concentrations of sodium nitroprusside (SNP) were added to the sperm suspension from 48 healthy fertile men, and the suspension was incubated in 1 x Earle at 37 degrees C for 1 hour. Progesterone was used to induce AR for 15, 30, 45 and 60 min, and then acid phosphatase (ACP) activity in the suspension before and after capacitation and at different time of AR was measured by p-nitrophenyl sodium phosphate assay. In the meantime, sperm motile parameters were assayed by CASA to observe sperm capacitation and AR.

RESULTSACP activity and sperm motile parameters increased in the 50 approximately 100 nmol/L NO concentration group, showed no significant variation in the 150 approximately 200 nmol/L group, and decreased in the 250 approximately 300 nmol/L group.

CONCLUSIONNO can facilitate sperm capacitation, AR and sperm motile parameters in low concentration and suppress them in high concentration. ACP activity assay of sperm is an objective and reliable method to evaluate sperm capacitation and AR in whole sperm population.

Acid Phosphatase ; metabolism ; Acrosome Reaction ; drug effects ; physiology ; Adult ; Dose-Response Relationship, Drug ; Humans ; Male ; Nitric Oxide ; physiology ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Sperm Capacitation ; drug effects ; physiology ; Sperm Motility ; drug effects ; physiology ; Spermatozoa ; enzymology

OBJECTIVETo investigate the influence of heat shock factor1 (HSF1) on gene expression of inflammatory mediators in RAW264.7 murine macrophage cells induced by burn serum.

METHODSSera were separated from blood of normal rats and rats with severe burns, and the recombinant vector pcDNA3. 1/HSF1 was constructed. RAW264.7 macrophages were divided into non-transfection group, vacant vector group (with burn and normal sera stimulation, respectively after vacant vector transfection) and recombinant vector group (with burn and normal sera stimulation, respectively after recombinant vector transfection). Some recombinant vector transfected macrophages without serum stimulation were prepared for the determination of HSF 1 expression with Western blotting. The mRNA expressions of TNF-alpha, HMGB 1 and IL-10 were determined with RT-PCR.

RESULTSThe cell line attained after recombinant vector transfection was comparatively stable,with partial activation of HSF 1. Burn sera markedly upregulated TNF-alpha, HMGB1 mRNA expression (0.910 +/- 0.100, 0.860 +/- 0.020, respectively), but downregulated IL-10 expression (0.430 +/- 0.010, respectively) in normal macrophages, while these genes maintained in a very low level in normal macrophages with normal serum stimulation . macrophages with recombinant vector transfection and burn serum stimulation could obviously inhibit the expression of TNF-alpha and HMGB 1, but enhance the IL-10 gene expression (0.130 +/- 0.100, 0.450 +/- 0.020 , 0.450 +/- 0.020, respectively )when compared with that with vacant vector transfection and burn serum stimulation (0.800 +/- 0.050, 0.880 +/- 0.030, 0.420 +/- 0.010, respectively).

CONCLUSIONHSF1 can inhibit the expression of some pro-inflammatory mediators in macrophages after a severe burns, indicating that appropriate upregulation of anti-inflammatory mediators might exert protective effects on the organism.

Animals ; Burns ; metabolism ; Cell Line ; DNA-Binding Proteins ; genetics ; Female ; Gene Expression ; HMGB1 Protein ; metabolism ; Heat Shock Transcription Factors ; Heat-Shock Response ; genetics ; Inflammation Mediators ; metabolism ; Interleukin-10 ; metabolism ; Macrophages ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Serum ; Transcription Factors ; genetics ; Transfection ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo find out the occurrence of cesarean section (CS) and to probe the factors associated with CS.

METHODSWomen with CS as "case group" and women without CS as "control group" were chosen in a case-control study.

RESULTSAmong 14 071 childbirth women, 6 421 had CS (case group) with the occurrence rate of 45.6% and 7 650 (54.4%) had normal delivery (control group). In comparison with the control group, the CS group had following several higher rates [with significant differences between case group and control group (P < 0.01)]: well-educated (78.9% vs 69.5%), white collar jobs (38.0% vs 32.3%), urban residents (79.1% vs 70.6%), high monthly income (>/= 500 Yuan) (81.0% vs 70.6%), of older age (>/= 25 years) (73.3% vs 63.0%), heavier baby weight (> 4 000 gram) (8.3% vs 2.9%), male babies (53.9% vs 51.4%), BMI of mother (> 24) (8.8% vs 4.8%), cephalopelvic disproportion (21.1% vs 0.9%), intrauterine asphysia (20.3% vs 6.7%), abnormality of force of labor (4.2% vs 2.7%), prolonged labor (2.9% vs 1.0%) and placenta previa (1.4% vs 0.4%). Our study also indicated that the higher the educational level was, the higher the rate of CS appeared; and the older the pregnant women was, the higher the rate of CS was. In CS group, over 70% primipara were over 24 years, and over 20% primipara had cephalopelvic disproportion and over 20% had intrauterine asphysia in CS group.

CONCLUSIONSAt present, the occurrence rate of cesarean section was rather high (45.6%) in China. The high rate of CS was more likely to associate not only with abnormal physiological/medical factors (eg. cephalopelvic disproportion, intrauterine asphysia, abnormality of force of labor, and prolonged labour, etc.), but also with some demographic factors as education, occupation, income and age, etc. It is necessary to take measures to reduce the unnecessary CS in China.

Adult ; Cesarean Section ; statistics & numerical data ; China ; Female ; Humans ; Logistic Models ; Pregnancy

OBJECTIVETo establish a diffuse large B-cell lymphoma (DLBCL)-mice model using human DLBCL cell line LY8, to investigate its characteristics of growth and to provide a model for in vivo study of DLBCL pathogenesis and treatment.

METHODSLY8 cells were injected subcutaneously into the right flank of nude mice. Harvested tumor tissues were cut into small pieces of 1.5 mm × 1.5 mm × 1.5 mm and implanted subcutaneously into nude mice. Tumor growth was visualized and the histologic characteristics were documented. Expression of LCA, CD20, CD79α, Ki-67, CD3, CD45RO, bcl-6, MUM-1, CD10 and bcl-2 were examined by using immunohistochemistry. IgH clonal rearrangement and status of three microsatellite loci (D14S68, D18S69, D20S199) in the xenografted tumor samples and the parental cell line LY8 were detected using PCR amplification followed by PAGE.

RESULTSThe subcutaneous xenograft DLBCL model was successfully established by using cell line LY8, and a stable growth was achieved up to the 9th generation. The tumor in each generation showed similar growth characteristics and the rate of subcutaneous tumor formation was 91.9% (114/124). The tumor growth was observed from the 2nd week after implantation, reaching 1.3 cm in major diameter at the 3rd week and 2.0 cm at the 4th week. The tumor had identical morphological characteristics with those of human DLBCL, and expressed LCA, CD20, CD79α, bcl-6, MUM-1, CD10 and bcl-2. The tumor of xenograft mice and cell line LY8 showed identical IgH rearrangement and microsatellite length.

CONCLUSIONSA human DLBCL bearing mouse model was successfully established. The mice model is similar to human counterpart with high stability and repeatability. Therefore, it provides an ideal animal model for in vivo studies of the biological characteristics and treatment of DLBCL.

Animals ; Antigens, CD20 ; metabolism ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Gene Rearrangement ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microsatellite Repeats ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

OBJECTIVETo explore the changes of CD(14)(+) monocyte human leucocyte antigen DR (HLA-DR) and their relationship with multiple organ dysfunction syndrome (MODS) in severe sepsis.

METHODSNinety-one patients with a definite diagnosis of severe sepsis in the intensive care unit (ICU) were included. CD(14)(+) monocyte HLA-DR levels were detected by flow cytometry on the first, 4th and 7th days of the study, and Marshall scores and prognosis on day 28 were evaluated.

RESULTSThirty-four patients died within 28 days following the onset with a mortality rate of 37.4%. Persistently lowered levels of HLA-DR were detected and significantly increased Marshall scores were found in the fatal cases at all the time points (P<0.001). In the surviving patients, the levels of HLA-DR were significantly increased (P<0.01) and Marshall scores were gradually decreased (P<0.001). During the observation period, the levels of HLA-DR decreased significantly as the number of dysfunctional organs and Marshall scores increased (P<0.001). The levels of HLA-DR were significantly increased in severe sepsis patients with 2-4 dysfunctional organs and Marshall score of 5-12 (P<0.05 or P<0.001). No changes in HLA-DR levels in severe sepsis patients with 5-6 dysfunctional organs and Marshall scores of 13-22. The levels of HLA-DR showed a significant inverse correlation to Marshall scores (r=-0.368, P<0.001).

CONCLUSIONIn patients with severe sepsis, persistent low CD(14)(+) monocyte HLA-DR levels predicts high mortality. The levels of HLA-DR are significantly correlated to the severity of organ dysfunction.

Adult ; Aged ; Aged, 80 and over ; Female ; HLA-DR Antigens ; metabolism ; Humans ; Lipopolysaccharide Receptors ; Male ; Middle Aged ; Monocytes ; immunology ; metabolism ; Multiple Organ Failure ; pathology ; Sepsis ; immunology ; metabolism

OBJECTIVETo compare the expression of nuclear matrix proteins (NMPs) in benign prostatic hyperplasia (BPH) epithelial cell line BPH-1 versus those in androgen-dependent human prostate cancer cell line LNCap and androgen-independent prostate cancer cell line PC-3.

METHODSWe isolated NMPs from the BPH-1, LNCap and PC-3 cell lines by 2-dimensional electrophoresis (2-DE), analyzed the differentially expressed proteins by matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS), and identified them by peptide mass fingerprint and database searching.

RESULTSWe successfully obtained well-resolved reproducible 2-DE patterns of NMPs in human prostate cancer cell lines, identified 12 differentially expressed NMPs including enzymes, regulatory proteins, RNA-binding protein and various other factors, 3 up-regulated and 9 down-regulated in prostate cancer cell lines.

CONCLUSIONThere are obvious differences in the expressions of NMPs between human prostate cancer cell lines and benign prostatic hyperplasia epithelial cell line.

Cell Line ; Cell Line, Tumor ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Male ; Nuclear Matrix-Associated Proteins ; metabolism ; Prostatic Hyperplasia ; metabolism ; Prostatic Neoplasms ; metabolism ; Proteome ; analysis ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization