To explore whether the Smad signal transduction pathway in human peritoneal mesothelial cells (HPMCs) influences the process of human peritoneal fibrosis stimulated by TGF-beta1. HPMCs were isolated from normal human omentum and 95% of the primary cultured cells was confirmed to be HPMCs. The third generation of cultured cells was stimulated by 5 ng/ml TGF-beta1. Immunohistochemistry, Western blotting, ELISA, and RT-PCR were employed to investigate the follows: the protein expression of p-Smad2/3 and its migration in HPMCs; the protein and mRNA expressions of SMAD 7 in cells; and the expressions of extracellular fibronectin (FN) protein, intracellular FN mRNA, as well as intracellular collenge-I (COL1) protein and mRNA. The protein expression of p-Smad2/3 in HPMCs obviously increased 15 min (29% p-Smad2/3-positive cells) after TGF-beta1 stimulation, peaking from 30 min (81% ) to 1 h (84%) and dropping after 2 h (37%); Meanwhile, p-Smad2/3 mainly distributed in cytoplasm at 15 min, concentrated in cell nucleus
To investigate the presence and significance of HBsAg, HBcAg, and HBV DNA in renal tissues from patients with HBV infection. The presences of HBsAg and HBcAg were measured in the renal tissues of 63 patients with HBV associated glomerulonephritis (HBV-GN), 20 patients with non-HBV associated glomerulonephritis (NHBV-GN), and 12 other renal patients with positive HBV markers (such as renal tuberculosis, renal calculi, renal cellular carcinoma, and renal atrophy, and so on) by immunocytochemistry. HBV DNA was detected in the renal tissues of those patients with positive immunocytochemic results by polymerase chain reaction (PCR). HBsAg and HBcAg were present with the positive rates of 76.2% and 42.9% respectively in the renal tissues of HBV-GN patients, which were significantly higher than those of NHBV-GN patients, but had no statistical difference compared with those of other renal patients with positive HBV markers. The positive expression rates of HBsAg and HBcAg in renal tissues showed no statistically sig
To explore the interaction between androgen receptor (AR) and silencing mediator for retinoid and thyroid hormone receptor (SMRT) and their interaction site. Methods We recombined and constructed AR, SMRT gene and gene fragments, in vitro translated 35S fusion proteins to investigate the relationship between AR and SMRT using transient transfection, mammalian two-hybrid test, GST pull-down assay, and indirect immunofluorescence staining. Results AR possessed an intrinsic transcriptional repression activity and AR interacted directly with SMRT. One interactive surface on AR was mapped to the ligand-binding domain (LBD), and the presence of DNA binding domain enhanced this interaction. The binding surface on SMRT was mapped to the carboxyl-terminal nuclear receptor interacting domain (ID), and mutation of the LXXXIXXXI/L corepressor motif within this domain interferred with the interaction. LBD domain on the AR can interact with ID2 motif on the SMRT.
To study the relationship between levels of activity of NF-kappaB p65, plasma soluble inter-cellular adhesion molecule-1, C-reactive protein on plaque stability, and different types of coronary heart disease. We measured the levels of plasma soluble inter-cellular adhesion molecule-1 and C-reactive protein by enzyme-linked immunosorbant assay and the activity of NF-kappaB p65 in peripheral blood lymocytes immunohistochemically. Compared with the stable angina and the control group, the baseline activity of NF-kappaB p65, sICAM-1 and C-reactive protein was significantly elevated in the acute myocardial infarction and the unstable angina (P 0.05). In all groups, C-reactive proteins were lowered at the review (P <0.01). The levels of activity of NF-kappaB p65, sICAM-1 and C-reactive protein are related to the plaque stability among different types of coronary heart disease. NF-kappaB p65, and sICAM-1 are not
To investigate the distribution of NMDAR1 and subunits (alpha1 and alpha3) of Immunohistochemistry and GABAA receptors in the cerebellar cortex of adult rhesus macaques. Nissl staining were used. (1) NMDAR1 immunoreactivity was prevalent in the endoplasma of Purkinje and dendrites of molecular layer, and moderate in glia, but was very weak in the stellate cells in the molecular layer and granule cells. In addition, cerebellar nuclei were also strongly stained; (2) The immunostaining pattern of subunit of GABAA alpha1 receptor was similar to that of NMDAR1, but more distinctive than NMDAR1 in stellate cells; (3) Immunoreactivity of subunit (alpha3) of GABAA receptor was also prevalent in the endoplasma of Purkinje cells and dendrites. The glia and the neurons of the cerebellar nucli were intensively stained. (4) The rates of Purkinje cells containing NMDAR1, and subunits of GABAA alpha1 and alpha3 were (90.0 +/- 2.1)%, (83.0 +/- 2.3)%, and (78.0 +/- 1.8)%, respectively. Conclusion NMDAR1 and subunits (alpha1 a
To determine the relationship between nitric oxide synthase (NOS) expression and Pgp/mdr-1. The expressions of iNOS, eNOS, and Pgp were detected by immunohistochemistry, and the expressions of iNOS gene and mdr-1 gene were detected by RT-PCR. NOS expression increased in patients with acute leukemia compared with that of the controls. There was some correlation between iNOS and Pgp/mdr-1 gene expression. NO may influcence multiple drug resistance in patients with acute leukemia.
To explore the significance and expression of transforming growth factor-beta1 (TGFbeta1) in cervical squamous carcinoma. The expression of TGFbeta1 in 31 cases of cervical squamous carcinomas were detected by SP immunohistochemical staining. TGFbeta1 expression was found in both cytoplasm and extracellular stroma. TGFbeta1 expression increased when the pathologic grade was getting worse and clinical stage was of later the in extracellular matrix and decreased in cytoplasm. The positive rate of TGFbeta1 in cytoplasm in the low grade group was lower than that in the high and middle grade (P 0.05. The decrease of TGFbeta1 expression may play an important
To determine the effect of atorvastatin on the activity of peripheral blood lymocyte nuclear factor-kappaB (NF-kappaB) and plasma soluble inter-cellular adhesion molecules-1 (sICAM-1) in acute coronary syndromes. Sixty-eight patients with acute coronary syndrome were randomly divided into atorvastatin therapeutic group (n = 37) and conventional therapeutic group (n = 31). Enzyme-linked immunosorbent assay was used to measure the plasma sICAM-1, and immunohistochemical method was used to measure the activity of NF-kappaB in the peripheral blood lymphocyte before and 12 weeks after the therapy in the two groups. Before the therapy, the level of NF-kappaB activity in the peripheral blood lymphocytes [(45.2 +/- 8.4)% vs (40.8 +/- 9.2)%, P > 0.05] and the plasma sICAM-1 [(357.2 +/- 84.5) ng/ml vs (365.5 +/- 91.3) ng/ml, P > 0.05] in the two groups had no significant difference. After 12 weeks of the therapy, in the conventional group the level of NF-kappaB activity in the peripheral blood lymphocytes [(40.8 +/- 9.
To investigate the differentiation of the rat bone marrow stromal cells (BMSCs) into neuron like cells which express glutamic acid decarboxylase (GAD) and choline acetyltransferase (ChAT). BMSCs of rats were flushed out with DMEM from femurs marrow and separated, and then planted in plastic flask. After being expanded and purified, the BMSCs were differentiated into neuron-like cells. Neurofilament (NF-200), microassociate protein-2 (MAP-2), neuron-specific nuclear protein (NeuN), nestin, gial fibrillary acidic protein (GFAP), GAD and ChAT were detected by immunohisitochemistry. The induced BMSCs exhibited neuronal morphological characteristics, and the expressions of NF-200, MAP-2, NeuN and nestin were positive, but the expression of GFAP was negative. About (3 +/- 0.8)% of the induced BMSCs expressed GAD, and (5 +/- 0.3)% expressed ChAT. BMSCs can differentiate into neuron like cells, and express GAD and ChAT partially.
To investigate the effect of HPV16E6 DNA vaccines on inducing the antitumor immune response to cervix carcinoma in vivo. We prepared pCB/HPV16E6 DNA vaccines to prevent and treat 615 modle mice transplanted E6 + U14 in vivo respectively. Survival and tumor sizes of all mice were recorded and their lymphnodes and lung's metastases were detected by microscopic observation. The cytotoxicity of the spleen T cells of those mice immuned with HPV E6 DNA vaccines against the E6 U14 and U14 was detected by MTT assay. HPV16 E6 DNA vaccines prevented the transplanted tumor growth effectively in inbred strain mice and cured some tumor-bearing mice (P < 0. 001). Pathological results showed that the number of lymphodes with metastases foci decreased in the preventing group (P < 0.05). HPV16 E6 DNA vaccines induced special and higher cytotoxicity T lymphocytes against the E6 + U14 (P < 0.001). HPV16 E6 DNA vaccines can induce antitumor immune protection of the mice against E6 + U14 cells, suggesting that the vaccines may be
To investigate the therapeutic effect of endoscopic variceal ligation (EVL) combined with splenectomy on patients with portal hypertension. The eliminating rate of varicose veins and recurrent bleeding rate were evaluated. Fifty-one paitents with cirrhosis of liver and portal hypertension were randomly divided into 2 groups. We treated 25 portal hypertensive patients using splenectomoy plus EVL. The therapeutic effect of pericardial devascularization in 26 patients with upper gastrointestinal bleeding due to portal hypertension served as the control. All patients were followed up for 12 months. There was no death due to the operation. All patients in the study group had no upper gastrointestinal rebleeding during the 12-month follow-up. The rebleeding rate in the control group was 11.5% (3/26). The varicose venis eliminating rate was 96% (24/25) in the study group and 50% (13/26) in the control group (P < 0.01). It showed no effect on portal hypertensive gastropathy in the study group, and pericardial devascu
To investigate a new therapy for endometriosis, which can inhibit the angiogenesis of ectopic endometrium. Xeogeneic endothelial cells (human umbilical vein endothelial cells, HUVECs) were used as vaccines, and a variety of controlled cells (including rat aortic endothelial cells and human fibroblast cells) were cultured, fixed and resuspended in PBS. Thirty-six Lewis rats with experimentally induced endometriosis were divided into 6 groups. Group A was treated with HUVECs (1 x 10(5)); Group B was treated with HUVECs (5 x 10(5)); Group C was treated with HUVECs (1 x 10(6)); Group D (a control group) was treated with PBS (0.5 ml); Group E (a control group) was treated with human fibroblast cells (5 x 10(5)); and Group F (a control group) was treated with rat aortic endothelial cells (5 x 10(5)). Microvessel desity (MVD), area and the proliferative index were deteced in different groups 4 weeks after the treatment. The effect of immune serum on rat endothelial cell proliferation in vivo was determined with meth
To investigate the changes of serum hepatocyte growth factor (HGF) in patients with acute renal failure (ARF) and to evaluate the clinical value. Serum HGF levels were determined by ELISA in 31 patients with ARF at the stages of oliguria, polyuria and convalescence and in 28 healthy controls. Compared with the controls, the HGF levels in the serum increased significantly in the ARF group at the stages of oliguria and polyuria, while the serum HGF returned to normal levels at the convalescence stage. There was significant positive correlation between the serum HGF and BUN or SCr in the ARF group. Serum HGF in ARF patients increased significantly and tended to rise at first but dropped afterwards, which reflected the course of renal injury and the recovery of renal epithelial cells.
To explore the changes of antioxidants in plasma and erythrocyte in epileptic patients. The levels of erythrocyte enzymes glutathione reductase (GR), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase and erythrocyte membrane lipid peroxidation (MDA), the percentage hemolysis, and plasma ceruloplasmin, vitamin A, vitamin C, vitamin E were measured in 39 controls, 32 epileptic patients and 26 follow-up patients. The levels of MDA, the percentage hemolysis, GSH-Px, CAT and plasma CER in the epileptic patients were significantly higher than those of the controls. While the levels of vitamin A, vitamin C, vitamin E, SOD and GR in the epileptic patients were markedly lower than those of controls. In the follow-up patients, the levels of GR, SOD, vitamin C and vitamin E were significantly higher than those of the epileptic patients. Free radicals and antioxidants play important roles in the etiology of epilepsy.
To investigate the regulatory effect of Chinese traditional medicine mixture (CTMM) on inflammatory response in rats with severe burn. One hundred and ten rats were randomly divided into 3 groups:scalded rats inflicted by 30% III degree scald were treated with CTMM and SD-Ag (CTMM group), scalded rats inflicted by 30% III degree scald were treated with SD-Ag alone (scalded group), and healthy rats were treated with SD-Ag (normal group). The serum contents of TNF-alpha, IL-beta, IL-6, IL-8, IL-4, and IL-10 in rats in the 3 groups were dynamically monitored. The serum contents of TNF-alpha, IL-1beta, IL-6, IL-8, IL4, and IL-10 evidently increased in both the CTMM and scalded groups. But the contents of pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6, and IL-8) in the CTMM group were much lower than those in the scalded group. However, the contents of anti-inflammatory cytokines (IL-4 and IL-10) in the CTMM group were much higher than those in the scalded group. CTMM has double-way regulatory effect on the
To investigate the effect of interleukin-12 (IL-12) on immune responses induced by pcDNA3 which expresses hepatitis B virus pan-S gene. BABL/c mice were immunized with pcDNA3 alone (control group), pcDNA3-PS alone, or co-immunized with pcDNA3-PS and pcDNA3-IL12. Six weeks after the immunization, sero anti-HBS antibody was examined, lymphocyte proliferation test and specific cytokine level were assayed. RESULTS The titer of anti-HBs antibody, the stimulation index of splenocytes, and cytokine level of IL-12 and IFN-gamma from the mice coimmunized with pcDNA3-PS and pcDNA3-IL12 were significantly higher than those of the mice immunized alone with pcDNA3-PS. Antibody level, SI, and cytokine level of the immunized mice were higher than those of the control group. IL-12 expression plasmid can act as immunological adjuvants for HBV pan-S gene vaccine.
To explore the mechanisms of C2C12 cell apoptosis induced by hydrogen peroxide (H2O2) which is inhibited by heat shock proteins. The expression of heat shock proteins in mouse embryonic myogenic cell line, C2C12 cell,was induced by heat shock response. C2C12 cell apoptosis induced by 0.5 mmol/L hydrogen peroxide (H2O2) was determined by Hoechst 33258 staining. The activities of caspase -3,8,9 were assayed by caspase colorimetric assay kit and Western-blotting. The release of cytochrome C from mitochondria was observed by Western-blotting of cell mitochondria and cytosol fractions. The expression of HSP70 and alphaB-crystallin in C2C12 cell significantly increased at 24 hour after the heat shock response. Heat shock response could inhibit the release of cytochrome c from mitochondria to cytoplasm,the activation of caspase -3,8,9 and the subsequent apoptosis induced by H2O2 in C2C12 cell. HSPs can inhibit the C2C12 cell apoptosis through interference with the activation of both mitochondrial and death receptor
To determine the relation between genotype of HBV and its clinical manifestation. Sera of 220 patients from Hunan Province of China, infected chronically with asymptomatic, mild, moderate, severe and fulminant hepatitis hepatitis B, were genotyped by PCR with genotype-specific primers. The results were statistically analyzed. Only genotype B (86.4%) and C (13.6%) were found. With the aggravations of patients' condition, genotype C was found more often (P < 0.05). The levels of alanine aminotransferase (ALT), total bilirubin (TBIL) and HBV-DNA of genotype C was higher than those of genotype B, but there was no statistical difference. However, the ALT elevation rates of genotype C (96.7%) were significantly higher than those of genotype B (75.2%) (P < 0.05). There was no statistical difference in HBeAg rate between genotype B and C in general, but in patients with chronic fulminant hepatitis or aged 21 approximately 30, the HBeAg rates of genotype C (35.0% and 50.0%) were significantly higher than those of geno