0.05).(3)The levels of CRH and DHEA-S in umbilical cord blood of PL(7.8?3.3)ng/L,and(514?295)?g/L,respectively and of TL (7.7?4.1)ng/L,and(483?207)?g/L,were higher than that in term not in labor(4.8?2.4)ng/L, and(360?80)?g/L,respectively(P 0.05).In PL,the level of CRH in umbilical cord blood and the expression of CRH mRNA in placentas and fetal membranes were correlated with each other(r=0.935 and 0.853,P

Angiotensin II ; pharmacology ; Blotting, Western ; Cells, Cultured ; Glomerular Mesangium ; cytology ; drug effects ; metabolism ; Humans ; I-kappa B Kinase ; NF-kappa B ; analysis ; Peptide Fragments ; pharmacology ; Protein-Serine-Threonine Kinases ; analysis

Objective To explore the inhibitory effect of rosiglitazone of peroxisome proliferator-activated receptor-?(PPAR?) agonist on aldosterone-induced mesangial cell(MC) proliferation.Methods Mouse primary MC were cultured and treated with aldosterone(100 nmol/L) in the presence or absence of rosiglitazone(1.0,2.5,5.0,10.0 ?mol/L).The incorporation of 3H-thymidine(3H-TdR) and cell count were used as the measure of MC proliferation.Cyclin D1 and cyclin A expression,PI3K and Akt phosphorylation were determined by Western blot analysis.Results 1.Aldosterone induced MC proliferation,as assessed by 3H-TdR incorporation and cell number,which were increased by 2.46-and 2.14-fold,respectively,in aldosterone-treated cells.Aldosterone-induced MC proliferation was inhibited by PPAR? agonist rosiglitazone in dose-dependent manner in mouse MC.2.Aldosterone induced cyclin D1 and cyclin A expression.Rosiglitazone reduced aldosterone-induced cyclin D1 and cyclin A expression in dose-dependent manner.3.Aldosterone induced PI3K/Akt activation in dose-dependent manner,incubation with 100 nmol/L aldosterone for 60 min,phosphorylation PI3K and Akt expression increased by above 3.0-fold.4.PI3K inhibitor LY294002 and Akt inhibitor significantly inhibited aldosterone-induced cyclin D1 and cyclin A expression.5.Rosiglitazone significantly inhibited aldosterone-induced PI3K/Akt activation,10 ?mol/L rosiglitazone almost completely blocked aldosterone-induced PI3K/Akt activation.Conclusion Rosiglitazone can block aldosterone-induced MC proliferation via inhibition of PI3K/Akt activation.

Objective To investigate the effect of c-Jun N-terminal kinase(JNK) specific inhibitor SP600125 on Angiotensin Ⅱ(AngⅡ)-induced transforming growth factor-?1(TGF-?1) and fibronectin (FN) expression in human mesangial cells (MC).Methods Human MC were isolated and cultured in vitro and were treated with AngⅡ in the presence or absence of JNK specific inhibitor SP600125.The protein was isolated or the supernate of medium was collected at the end of experiment.JNK,extracellular signal-regulated kinase(ERK1/2),and p38 mitogen-activated protein kinase(MAPK) activity were determined by Western blot method.TGF-?1 and FN were determined by enzyme linked immunosorbent assay(ELISA).Results SP600125 inhibited AngⅡ-induced Ser63 phosphorylation of c-Jun in a concentration-dependent manner,and JNK activity was reduced by 75% at 10 ?mol/L and by 90% at 20 ?mol/L.SP600125 had no effect on AngⅡ-induced ERK1/2 and p38 activity.TGF-?1 and FN protein were constitutively produced in MC,and production was significantly stimulated for 8 to 48 h after addition of AngⅡ.Preincubation of cells with SP600125(20 ?mol/L) significantly inhibited AngⅡ-induced TGF-?1 and FN production during this time period.SP600125 inhibited AngⅡ-induced production of TGF-?1 and FN in a concentration-dependent manner.Conclusion SP600125 inhibited AngⅡ-induced JNK activation and TGF-?1 and FN expression in human MC and may serve as the novel approach for the treatment of patients with chronic kidney disease.

0.05)between the two groups.Conclu- sion The long-term outcomes of e-CHB is not markedly different compared with that of e+CHB.

OBJECTIVETo observe the changes of the cytokines following stereotactic irradiation for hepatocarcinoma with cirrhosis in rabbits.

METHODSSixteen rabbits with liver cirrhosis and hepatocarcinoma (experimental group) were randomized into two equal groups to receive stereotactic irradiation at single dose of 20 and 30 Gy, respectively. Eight rabbits with hepatocarcinoma (control group) were divided into two equal groups and treated in identical manner. All the rabbits were killed 3 weeks after irradiation, and EV two-step method was used to observe the cytokine changes of proliferating cell nuclear antigen (PCNA) and glutathione S-transferase pi (GST-pi) after irradiation.

RESULTSAfter irradiation, PCNA and GST-pi expression showed significant difference in the adjacent liver tissue between the experimental and control rabbits with irradiation at 20 Gy (P=0.010), but not with the irradiation dose of 30 Gy (P=1.000). Irradiation at different doses resulted in significant difference in the cytokine expression in the experimental rabbits (P=0.010). In the liver tissue exposed to irradiation, different irradiation doses resulted in significant difference in PCNA and GST-pi protein expression (P=0.010).

CONCLUSIONSFor hepatocarcinoma with cirrhosis in rabbits, radiation at the single dose of 30 Gy produces better response than 20 Gy, and PCNA and GST-pi may serve as good indexes for evaluating the therapeutic effect.

Animals ; Glutathione S-Transferase pi ; biosynthesis ; Immunohistochemistry ; Liver Cirrhosis, Experimental ; metabolism ; radiotherapy ; Liver Neoplasms, Experimental ; metabolism ; radiotherapy ; Male ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Rabbits ; Radiotherapy Planning, Computer-Assisted ; methods ; Radiotherapy, Conformal ; Random Allocation

BACKGROUNDOur previous study has confirmed that one bout of exhaustion (Ex) can cause hippocampus neurocyte damage, excessive apoptosis, and dysfunction. Its initial reason is intracellular calcium overload in hippocampus triggered by N-methyl-D-aspartic acid receptor (NMDAR) over-activation. NMDAR activation can be suppressed by γ-aminobutyric acid (A) receptor (GABAAR). Whether GABAAR can prevent intense exercise-induced hippocampus apoptosis, damage, or dysfunction will be studied in this study.

METHODSAccording to dose test, rats were randomly divided into control (Con), Ex, muscimol (MUS, 0.1 mg/kg) and bicuculline (BIC, 0.5 mg/kg) groups, then all rats underwent once swimming Ex except ones in Con group only underwent training. Intracellular free calcium concentration ([Ca2+]i) was measured by Fura-2-acetoxymethyl ester; glial librillary acidic protein (GFAP) and synaptophysin (SYP) immunofluorescence were also performed; apoptosis were displayed by dUTP nick end labeling (TUNEL) stain; endoplasmic reticulum stress-induced apoptosis pathway was detected by Western blotting analysis; Morris water maze was used to detect learning ability and spatial memory.

RESULTSThe appropriate dose was 0.1 mg/kg for MUS and 0.5 mg/kg for BIC. Ex group showed significantly increased [Ca2+]i and astrogliosis; TUNEL positive cells and levels of GFAP, B cell lymphoma-2 (Bcl-2) associated X protein (Bax), caspase-3, caspase-12 cleavage, CCAAT/enhancer binding protein homologous protein (CHOP), and p-Jun amino-terminal kinase (p-JNK) in Ex group also raised significantly compared to Con group, while SYP, synapse plasticity, and Bcl-2 levels in Ex group were significantly lower than those in Con group. These indexes were back to normal in MUS group. BIC group had the highest levels of [Ca2+]i, astrogliosis, TUNEL positive cell, GFAP, Bax, caspase-3, caspase-12 cleavage, CHOP, and p-JNK, it also gained the lowest SYP, synapse plasticity, and Bcl-2 levels among all groups. Water maze test showed that Ex group had longer escape latency (EL) and less quadrant dwell time than Con group; all indexes between MUS and Con groups had no significant differences; BIC had the longest EL and least quadrant dwell time among all groups.

CONCLUSIONSActivation of GABAA R could prevent intense exercise-induced synapses damage, excessive apoptosis, and dysfunction of hippocampus.

Animals ; Apoptosis ; physiology ; Body Weight ; physiology ; Endoplasmic Reticulum Stress ; physiology ; Hippocampus ; metabolism ; Male ; Physical Exertion ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, GABA ; genetics ; metabolism ; Synapses ; pathology

OBJECTIVETo explore the role of exogenous connective tissue growth factor (CTGF) in the collagen III synthesis of human renal tubular epithelial cell line HK2 in vitro.

METHODSCultured HK2 cells were randomly assigned to three groups: placebo-control, low-dose CTGF-treated (2.5 ng/mL) and high-dose CTGF-treated groups (20 ng/mL). Cell morphological changes were observed under an inverted microscope. Collagen III alpha mRNA expression was detected using RT-PCR. Immunohistochemistry staining was used to assess the levels of intracellular collagen III alpha protein.

RESULTSAfter 48 hrs of low- or high- dose CTGF treatment, the appearances of HK2 cells were changed from oval to fusiform. High-dose CTGF treatment increased collagen III alpha mRNA expression (0.4461+/-0.0274 vs 0.2999+/-0.0115; P<0.05) as well as the protein expression of collagen III alpha (0.4075+/-0.0071 vs 0.3503+/-0.0136; P<0.05) compared with the placebo-control group.

CONCLUSIONSCTGF can induce morphological changes of human renal tubular epithelial cells in vitro. High concentration of CTGF may increase the synthesis of collagen III alpha.

Cells, Cultured ; Collagen Type III ; biosynthesis ; genetics ; Connective Tissue Growth Factor ; Epithelial Cells ; drug effects ; metabolism ; Humans ; Immediate-Early Proteins ; pharmacology ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Kidney Tubules ; drug effects ; metabolism ; RNA, Messenger ; analysis

AIMTo investigate the distribution of cation channel of sperm 1 (CATSPER1) protein and the presence of CATSPER1 mRNA in human testis and ejaculated spermatozoa. The influence of anti-human CATSPER1 antibody upon human sperm motility was used to evaluate the function of human CATSPER1 and to estimate its possible use as a target for immunocontraception.

METHODSHuman ejaculated sperm from normozoospermic donors (n = 12) and liquid nitrogen frozen human testis were used for the study of mRNA and protein expression of CATSPER1 by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, respectively. Spermatozoa from normozoospermic donors (n = 12) were individually processed using a swim-up procedure and were then incubated with CATSPER1 antibody at final concentrations of 20, 4 and 0.8 microg/mL. After 1, 2 and 6 h incubation, progressive motility and fast progressive motility were measured by means of computer-assisted semen analysis.

RESULTSCATSPER1 transcript was detected in both human testis and each human ejaculated semen sample. CATSPER1 protein expressed in the membrane of spermatid and was localized in the principal piece of the sperm tail. The application of CATSPER1 antibody at all concentrations significantly inhibited both progressive motility and fast progressive motility after 1, 2 and 6 h incubation, and significant dose-dependent changes were observed.

CONCLUSIONCATSPER1 is meiotically and post-meiotically expressed in human testis tissue. CATSPER1 mRNA in human ejaculated spermatozoa could be a more feasible target for study and infertility screening than testis biopsy. In addition, our results suggest that human CATSPER1 could be a possible target for immunocontraception.

Antibodies ; Calcium Channels ; genetics ; immunology ; Ejaculation ; Humans ; Male ; Meiosis ; Protein Biosynthesis ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Semen ; physiology ; Sperm Motility ; immunology ; physiology ; Spermatozoa ; cytology ; physiology ; Testis ; cytology ; physiology ; Transcription, Genetic

OBJECTIVESepsis remains a serious clinical problem because of high morbidity and mortality. The importance of Toll-like receptors (TLRs) for the induction of immune responses against sepsis was demonstrated in humans. The present study aimed to probe the gene polymorphisms of TLR4 (Asp299Gly and Thr399Ile) and TLR2 (Arg753Gln) in patients with sepsis among Chinese Han children in Wenzhou, and investigate the correlation with sepsis.

METHODThis study was conducted as a case-control study. Using polymerase chain reaction and DNA sequencing, gene polymorphisms of TLR4 (Asp299Gly and Thr399Ile) and TLR2 (Arg753Gln) in 59 children with sepsis, 38 children with severe sepsis (including 20 septic shock) and 57 healthy controls were analyzed. Hardy-Weinberg method of statistics was used to compare the frequency of genotypes alleles among three groups.

RESULTThe mutant genotypes of TLR4 gene (Asp299Gly and Thr399Ile) were not found among sepsis, septic shock and control groups. In severe sepsis group, the Arg753Gln TLR2 polymorphism occurred in 2 out of 38 severe sepsis patients and both of the subjects with the TLR2 Arg753Gln polymorphism had fatal staphylococcal infections.

CONCLUSIONTLR4 gene (Asp299Gly and Thr399Ile) polymorphisms may not be correlated with susceptibility to sepsis among Chinese Han children in Wenzhou. The fact that only 2 out of 38 severe sepsis patients had Arg753Gln TLR2 polymorphism suggests that a larger sample size is needed because of the rarity of the TLR2 allele among Chinese Han children in Wenzhou.

Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; China ; Female ; Genetic Predisposition to Disease ; Humans ; Male ; Polymorphism, Single Nucleotide ; Sepsis ; ethnology ; genetics ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics