Aim The expression of NF-?B activation and the effect of antioxidant (N- acetylcysteine, NAC) on NF-?B activity in human airway epithelial cell was assessed. Methods Using the TNF-?,the airway epithelial cell strains of normal subject(16HBE) and tumor patient (H292) was activated and using Western-Blot and ELISA the expression of NF-?B and IL-8 were detected. Results It was found that the activity of NF-?B could be stimulated by the TNF-? and increase with the amount of TNF-? with the peak occurring at 2 to 4 hours after stimulation and then decreasing at six hours. At the same time, the level of IL-8 was elevated, but decreased with inhibition of NF-?B activity by NAC, that means the action of NAC has a dose-dependent effect. Conclusion NAC not only blocks the signal transmission activated by NF-?B, but also anticipates the transcription modulation of expression of many cell factors and inflammatory mediums. It suggests that NAC may play a role in the anti-inflammatory treatment of respiratory diseases.

The properties of polyethyleneimine-cholesterol cationic lipopolymer (PEI-Chol) as gene carries and its gene transfer efficiency in vitro with lipid microbubbles were presented in this paper. PEI-Chol lipopolymer was synthesized by linking cholesterol chloroformate to the amino groups of branched poly(ethyleneimine) (PEI) of 1 800. The structure and molecular weight of PEI-Chol were confirmed by IR, 1H NMR and MADI-TOF-MS (matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry), respectively. The average molecular weight of PEI-Chol was approximately 2 000. The gene delivery system of bubble/PEI-Chol/DNA was constructed by mixed PEI-Chol/pDNA (N/P 10:1) complexes with lipid microbubbles (2-8 microm) which were prepared by DPPC, DSPE-PEG2000 and perfluoropropane with the reverse phase evaporation technique. pEGFP-Cl (enhanced green fluorescent protein) was used as report gene to investigate the DNA condensing ability of PEI-Chol lipopolymer by agarose gel electrophoresis. And their cytotoxicity and in vitro transfer efficiency of different complexes were compared with each other in A549 and MCF-7. The results indicated PEI-Chol lipopolymer can condense plasmid DNA when N/P ratio upto 4, PEI-Chol complexes and bubble/PEI-Chol/DNA complexes were nontoxic to A549 and MCF-7 when formulated at the N/P ratio of 10/1 as determined by MTT assay. This bubble/PEI-Chol/DNA delivery system provided good transfer efficiency with other desirable characteristics such as against-precipitation of plasma proteins. In conclusion, bubble/PEI-Chol/DNA complex is a novel non-viral gene delivery system.
1,2-Dipalmitoylphosphatidylcholine ; chemistry ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cell Survival ; Cholesterol ; chemistry ; Contrast Media ; DNA ; chemistry ; genetics ; Female ; Fluorocarbons ; chemistry ; Gene Transfer Techniques ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Lipids ; chemistry ; Lung Neoplasms ; pathology ; Microbubbles ; Phosphatidylethanolamines ; chemistry ; Plasmids ; Polyethylene Glycols ; chemistry ; Polyethyleneimine ; chemistry ; Transfection ; methods

OBJECTIVETo observe the effects of Epimedium total Flavonoids Phytosomes on preventing and treating bone-loss of the castrate osteoporosis rat model.

METHODThe osteoporosis model was established with 4-month-odl panther's rats, their ovaries on both sides castrated. Dual energy X-ray scanning was used to determine the bone density, and immunity and ELASA were used to assay concentration of estradiol and IL-6 in serum respectively, then determine their effect.

RESULTThe BMP and E2 of high dosage group nilestriol group and normal group are higher than those of model group (P < 0.01), while their content of IL-6 is apparently lower than that of model group(P < 0.01).

CONCLUSIONThe osteoporosis model was established successfully and the using of EFP can improve the bone density, enhance E2 level and decrease the IL-6 concentration in serum.

Animals ; Bone Density ; drug effects ; Drug Carriers ; Epimedium ; chemistry ; Estradiol ; blood ; Female ; Flavonoids ; administration & dosage ; isolation & purification ; pharmacology ; Interleukin-6 ; blood ; Osteoporosis ; blood ; drug therapy ; prevention & control ; Ovariectomy ; Phospholipids ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influence of stromal cells on the Kallikrein 7 (KLK7) expression of epithelial cells in benign prostate hyperplasia (BPH).

METHODSWe constructed a stromal-epithelial co-culture model after separating the two types of cells from BPH tissues and identifying them by cell morphology and chemiluminescent microparticle immunoassay (CMIA). The expression of KLK7 mRNA was detected by RT-PCR in the epithelial cells with or without the stromal cells, and that of the KLK7 protein (hK7) determined by Western blot.

RESULTSStromal and epithelial cells were successfully separated and identified, and a stromal-epithelial co-culture model successfully established. RT-PCR showed that the mRNA expression of the KLK7 gene was higher in the epithelial cells co-cultured with stromal cells than in the epithelial cells alone, and the gray value of KLK7 to GAPDH was 1.41 +/- 0.041 in the former and 1.78 +/- 0.10 in the latter (P < 0.01). The results of Western blot were consistent with those of RT-PCR.

CONCLUSIONStromal cells can suppress the expression of the KLK7 gene in the epithelial cells in BPH. KLK7 may be involved in the change of epithelial cells stimulated by stromal cells.

Cells, Cultured ; Humans ; Kallikreins ; metabolism ; Male ; Prostate ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; Stromal Cells ; metabolism

Objective To make a reasonable selection from internal fixation Methods for treating femoral neck fracture, namely the fixation angle of a single screw, the number of screws and the combination mode of screws, so as to obtain the optimal stability of fracture reduction. Methods Based on the DICOM data and Lindon mode, a three-dimensional finite element model of femoral neck fracture fixation was built including different angles, numbers and combination modes of the screws. The model was attributed to nonlinear material properties based on the relationship between the property parameters of bone materials and the gray value of CT images, and loaded under the simulation of physiological loads. Results Corresponding to different angles of the fracture surface, the optimal fixation angle of the single screw was 65°and 70°in clinic. The more the number of screws, the better the effect of fixation, when the surgical condition was permitted. The inverted triangle placement was better than the triangle placement in case of three-screw fixation, while the triangle placement was superior to two-screw fixation, but the decision on the placement of two-screw fixation in flat form or diagonal form depended on the angle of the fracture surface. Conclusions Different angles of the fracture surface have significant impacts on the effect of fixation, and the loading simulation process on the three-dimensional model can provide a feasible way to the study of the fixation for the femoral neck fracture.

The fundamental event of cancer invasion and metastasis is the complicated interaction of cancer cells with host cells, in which event, a number of proteases and their inhibitors are involved. Matrix metalloproteinases are the potent proteases in degrading the basement membrane and extra cellular matrix and are inhibited by specific endogeneous inhibitors, tissue inhibitors of metalloproteinases-1(TIMP-1) and TIMP-2. The expression of mRNA for TIMP-1 and -2 was investigated by Northern blot analysis in specimens taken from 27 patients with primary gastric adenocarcinoma; 25 samples from the primary site, six from the metastatic lymph nodes and two from the peritoneal fluids. The expression for TIMP-1 and -2 was compared in primary gastric cancer tissues, metastatic lymph nodes and normal gastric mucosae. TIMP-1 mRNA was overexpressed in 24 (96%) out of 25 primary cancer tissues compared with the paired normal mucosae, while TIMP-2 was in 10 (40%). In six specimens of metastatic lymph nodes, TIMP-1 and -2 were overexpressed in 6 (100%) and 4 (67%) specimens, respectively. Of two specimens prepared from the peritoneal fluids, all specimens overexpressed TIMP-1 compared with the those of primary cancer tissues, while one (50%) specimen overexpressed TIMP-2. Immunohistochemical staining was done to investigate the localization of TIMP-1 and -2, demonstrating that the immunoreactivity for TIMP-1 and -2 was clearly detected in the cytoplasm of the stromal cells. These results suggest that both TIMP-1 and -2 are overexpressed by stromal cells in most of primary and some metastatic gastric cancer tissues and that TIMP-1 and TIMP-2, produced by stromal cells, may play an important role in inhibiting the proteolytic activity of matrix metalloproteinases originated from cancer cells, in gastric cancer.
Adenocarcinoma/*enzymology ; Blotting, Northern ; Glycoproteins/*biosynthesis/genetics ; Human ; Proteins/*biosynthesis/genetics ; RNA, Messenger/biosynthesis ; Stomach Neoplasms/*enzymology ; Support, Non-U.S. Gov't ; Tissue Inhibitor of Metalloproteinases ; Tissue Inhibitor-of Metalloproteinase-2

The fundamental event of cancer invasion and metastasis is the complicated interaction of cancer cells with host cells, in which event, a number of proteases and their inhibitors are involved. Matrix metalloproteinases are the potent proteases in degrading the basement membrane and extra cellular matrix and are inhibited by specific endogeneous inhibitors, tissue inhibitors of metalloproteinases-1(TIMP-1) and TIMP-2. The expression of mRNA for TIMP-1 and -2 was investigated by Northern blot analysis in specimens taken from 27 patients with primary gastric adenocarcinoma; 25 samples from the primary site, six from the metastatic lymph nodes and two from the peritoneal fluids. The expression for TIMP-1 and -2 was compared in primary gastric cancer tissues, metastatic lymph nodes and normal gastric mucosae. TIMP-1 mRNA was overexpressed in 24 (96%) out of 25 primary cancer tissues compared with the paired normal mucosae, while TIMP-2 was in 10 (40%). In six specimens of metastatic lymph nodes, TIMP-1 and -2 were overexpressed in 6 (100%) and 4 (67%) specimens, respectively. Of two specimens prepared from the peritoneal fluids, all specimens overexpressed TIMP-1 compared with the those of primary cancer tissues, while one (50%) specimen overexpressed TIMP-2. Immunohistochemical staining was done to investigate the localization of TIMP-1 and -2, demonstrating that the immunoreactivity for TIMP-1 and -2 was clearly detected in the cytoplasm of the stromal cells. These results suggest that both TIMP-1 and -2 are overexpressed by stromal cells in most of primary and some metastatic gastric cancer tissues and that TIMP-1 and TIMP-2, produced by stromal cells, may play an important role in inhibiting the proteolytic activity of matrix metalloproteinases originated from cancer cells, in gastric cancer.
Adenocarcinoma/*enzymology ; Blotting, Northern ; Glycoproteins/*biosynthesis/genetics ; Human ; Proteins/*biosynthesis/genetics ; RNA, Messenger/biosynthesis ; Stomach Neoplasms/*enzymology ; Support, Non-U.S. Gov't ; Tissue Inhibitor of Metalloproteinases ; Tissue Inhibitor-of Metalloproteinase-2

BACKGROUNDSevere acute respiratory syndrome coronavirus (SARS-CoV) is a newly emerging virus that gives rise to SARS patients with high rates of infectivity and fatality. To study the humoral immune responses to SARS-CoV, the authors evaluated IgG and IgM specific antibodies in patients' sera.

METHODSTwo methods, enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent assay (IFA), were used to detect specific serum IgG and IgM against SARS-CoV in 98 SARS patients and 250 controls consisting of patients with pneumonia, health-care professionals and healthy subjects. The serum antibody profiles were investigated at different times over one and a half years in 18 of the SARS patients.

RESULTSThe sensitivity and specificity of ELISA for detecting IgG against SARS-CoV were 100.0% and 97.2% and for IgM 89.8% and 97.6% respectively; the figures using IFA for IgG were 100.0% and 100.0% and for IgM 81.8% and 100.0% respectively. During the first seven days of the antibodies trace test, no IgG and IgM were detected, but on day 15, IgG response increased dramatically, reaching a peak on day 60, remaining high up to day 180 and decreasing gradually until day 540. On day 15, IgM was detected, rapidly reached a peak, then declined gradually until day 180 when IgM was undetectable.

CONCLUSIONThe detection of antibodies against SARS virus is helpful in the clinical diagnosis of SARS.

Adult ; Aged ; Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Male ; Middle Aged ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVE: To investigate the difference of Pax2 and P53 expressions in children with primary nephritic syndrome (PNS) and the effect of Pax2 on glucocorsteroid (GC)-resistance. METHODS: Renal Pax2 and P53 expressions in children with PNS (40 patients) were detected by immunohistochemistry. A semiquantitative score was used to evaluate the injury degree of the glomeruli and the tubulointerstitium, and correlation analysis was done among Pax2, P53 and pathologic score. RESULTS: Pax2 and P53 expressions were not found in the control group. Pax2 expression of renal tubule epithelia exsisted in children with PNS and there was weak or no expression of Pax2 in the podocytes. Pax2 expressions in the proximal tubule and the distal tubule in the GC-resistant group were more intense than those in the GC-intensive group (P <0.01). The more the Pax2 expression in the tubule, the more abnormal structure such as dilation and atrophy. Pax2 expression in the tubule epithelia was positively correlated with pathologic score of tubulointerstitium (P < 0.01). There was no P53 expression in the GC-intensive group, but there exsisted P53 expression in parts of the patients from the GC-resistant group, mainly distributing in the renal tubular epithelia. P53 expression was positively correlated with P53 expression and the pathologic score of tubulointerstitium (P < 0.01). CONCLUSION Over-expression of Pax2 in the renal tubule epithelia may improve P53 expression to a certain degree, which may aggravate the lesion of the renal tubule. It may be one of the mechanisms resulting in GC-resistant in children with PNS.
Adolescent ; Child ; Child, Preschool ; Drug Resistance ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Nephrotic Syndrome ; drug therapy ; metabolism ; PAX2 Transcription Factor ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVE: To investigate the efficacy and adverse effect of mycophenolate mofetil (MMF) in the treatment of frequently relapsing nephrotic syndrome in children. METHODS: The study population consisted of 37 children (24 simple nephrotic syndrome and 13 nephritis-type syndrome) suffering from frequently relapsing nephrotic syndrome. Patients received 20-30 mg/(kg d) of MMF in conjunction with 1 mg/(kg d) prednisone for 3-6 months. RESULTS: Out of 24 patients suffered from simple nephrotic syndrome, 17 patients (70.8%) with complete relief, 4 patients (16.7%) with partial relief and 3 patients (12.5%) with non-relief, whereas out of 13 patients suffered from nephritis-type syndrome 6 patients (46.2%) with complete relief, 3 patients (23.1%) with partial relief and 4 patients (30.7%) with non-relief. Eight patients with Minimal Change Disease (MCD) achieved complete relief. Of 23 patients with Mesangial Proliferative Glomerulonephritis (MsPGN) or Membranoproliferative Glomerulonephritis (MPGN), complete relief was observed in 17 patients (73.9%), partial relief in 4 patients (17.4%) and non-relief in 2 patients. CONCLUSION These Results suggest that MMF has better efficacy against simple renal disease than against nephritis-type syndrome, and MMF may be more suitable for the treatment of frequently relapsing nephrotic syndrome characterized by proliferative lesions.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Male ; Mycophenolic Acid ; adverse effects ; analogs & derivatives ; therapeutic use ; Nephrotic Syndrome ; drug therapy ; Recurrence ; Treatment Outcome