OBJECTIVETo identify the expression and distribution of macrophage colony stimulating factor (M-CSF) in brains of PDAPPV717I transgenic mice.

METHODSWe detected the expression and distribution of M-CSF mRNA in brains of PDAPPV717I transgenic mice by using hybridization in situ and immunohistochemical staining.

RESULTSExpression of M-CSF mRNA was significantly higher in brains of PDAPPV717I transgenic mice than that in non-transgenic mice, and M-CSF mRNA in brain was mainly produced by reactive astrocytes.

CONCLUSIONThe results indicate that astrocytes play an important role in the onset/development of neuropathology of Alzheimer's disease.

Alzheimer Disease ; genetics ; metabolism ; Animals ; Brain ; metabolism ; Female ; Immunohistochemistry ; In Situ Hybridization ; Macrophage Colony-Stimulating Factor ; biosynthesis ; genetics ; Male ; Mice ; Mice, Transgenic ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo examine the changes of expressions of orexin A, orexin receptor-1 (OX1R), prepro-orexin (Prepro-OX) mRNA, OX1R mRNA and ob-R of hypothalamus in rats with chronic renal failure (CRF).

METHODSSixty-two male Wister rats weighing 200-250 g were divided into three groups, including group 1 (normal, n = 5), group 2 (sham-operated, n = 25) and group 3 (CRF, n = 32). Hypothalamus orexin A was assayed by radioimmunoassay. Serum leptin was assayed by enzyme linked immunosorbent assay. The expression of Prepro-OX mRNA and OX1R mRNA of hypothalamus were measured by reverse transcription polymerase chain reaction, and expression of orexin A, OX1R and ob-R by immunohistochemistry. Automatic biochemical analyzer was used to measure the serum creatinine.

RESULTSHypothalamus orexin A levels were negatively correlated (r = -0.63, P < 0.001) with serum leptin levels in the rats. The expression of hypothalamus Prepro-OX mRNA in CRF rats was significantly lower than that of sham-operation at week 12 (P < 0.01). Hypothalamus Prepro-OX mRNA levels were negatively correlated (r = -0.81, P < 0.001) with the levels of serum leptin and serum creatinine (r = -0.68, P < 0.05) in the rats at week 12. The expression of hypothalamus OX1R mRNA in CRF rats was lower than that of sham-operation at week 12 (P > 0.05). Specific immunoreactivity for orexin A was present in perikeryon of the hypothalamus neuron. Specific OX1R-like immunoreactivity was observed in some nerve fibres. Specific immunoreactivity for ob-R was present in membranes of the hypothalamus neuron. Hypothalamus neurons of orexin A-like specific immunoreactivity in CRF rats were significantly fewer than those in shamoperated rats at week 8. Hypothalamus neurons of OX1R-like specific immunoreactivity in CRF rats were similar to those in sham-operated rat at week 8. Hypothalamus neurons of ob-R-like specific immunoreactivity in CRF rats were significantly more than those in sham-operated rats at week 8.

CONCLUSIONSThe lower hypothalamus orexin A levels may be induced by high serum leptin level in CRF rats. The lower expression of hypothalamus Prepro-OX mRNA in CRF rats may be one of the main causes inducing lower hypothalamus orexin A. The expression of OX1R in hypothalamus neurons is somewhat reduced and the expression of ob-R in hypothalamus neurons is somewhat raised in CRF rats. These remain to be studied further.

Animals ; Carrier Proteins ; genetics ; metabolism ; Hypothalamus ; metabolism ; Intracellular Signaling Peptides and Proteins ; Kidney Failure, Chronic ; metabolism ; Leptin ; genetics ; metabolism ; Male ; Neuropeptides ; genetics ; metabolism ; Neurotransmitter Agents ; genetics ; metabolism ; Orexin Receptors ; Orexins ; Protein Precursors ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Cell Surface ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; Receptors, Leptin ; Receptors, Neuropeptide ; genetics ; metabolism

OBJECTIVETo verify the binding of p53 to p21WAF1/CIP1 gene promoter and detect its binding to hsp90 beta gene promoter in vivo.

METHODSChromatin immunoprecipitation and PCR analysis were used to measure specific gene regulation sequence and Western blot analysis to investigate p53 protein.

RESULTSThe p53 binding sequences on the promoters of p21WAF1/CIP1 and hsp90 beta gene were found in the p53 antibody immunoprecipitated DNA fragments and p53 was detected in the immunoprecipitated samples.

CONCLUSIONSp53 binds to promoters of p21WAF1/CIP1 and hsp90 beta gene in vivo, and regulates the expression of the two genes.

Binding Sites ; Chromatin ; genetics ; isolation & purification ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; genetics ; metabolism ; Gene Expression Regulation ; HSP90 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Polymerase Chain Reaction ; methods ; Precipitin Tests ; methods ; Promoter Regions, Genetic ; Protein Binding ; Transcription, Genetic ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; physiology

OBJECTIVETo explore the characteristics of protective immunity against Plasmodium yoelii (P.y.) infection by asexual blood-stages cellular vaccine.

METHODSThe particulate vaccines were constructed by saponin or double-distilled-water lysed parasitic red blood cells and inoculated into BALB/c mice by intraperitoneal injection (i.p.). Each group was challenged by the lethal erythrocytic P.y. parasites, and then their parasitemia and survival rates were detected. Expressions of interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) were detected by RT-PCR. ELISA showed the serum antibodies against the malaria challenge and their-subclasses. Special membrane protein was recognized by immunofluorescence assay.

RESULTSThe vaccination with saponified erythrocytic parasites protected the immunized mice against P.y. challenge, while double-distilled-water lysed vaccine did not (P < 0.01). This protection was characterized by the increase of both IFN-gamma/IgG2a and IL-4/IgG1. Meanwhile, MHC class I alpha chain molecule was recognized on the membrane of infected-erthythrocyte.

CONCLUSIONSaponified P.y. asexual blood-stage cellular vaccine has a significantly high protective immunity against this lethal P.y. malaria, and the immunity may be associated with the expression levels of IgG2a and IFN-gamma. MHC class I alpha chain on infected erythrocytes may play an important role in the successful immunization.

Animals ; Antibodies, Protozoan ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Histocompatibility Antigens Class I ; blood ; Immunoglobulin G ; blood ; Malaria ; prevention & control ; Malaria Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Plasmodium yoelii ; immunology ; Random Allocation ; Vaccination

OBJECTIVETo evaluate the animal model of the multidrug resistant glioma cell line C6/adr for further in vivo studies.

METHODSThe rat glioma cells C6 and multidrug resistance cells C6/adr were cultured in vitro and implanted into the brain of S-D rats. After implantation, all these animals were examined continually with magnetic resonance imaging (MRI) and histological examination. The growth procedure of intracranial implanted glioma and the survival span of the animal model were evaluated. The statistical analysis was made between the survival data of the two cell lines.

RESULTSThe symptoms of intracranial hypertension did not occur until 4 weeks after inoculation. The MRI findings of the implanted glioma in the rat brain were much earlier than the abnormal behavior observed. Pathological results after inoculation demonstrated the MRI findings. The two cell lines had similar growth characteristics and no significant differences in survival times.

CONCLUSIONThese results suggest that by means of MRI and histology the growth procedure of the implanted glioma in vivo be successfully observed. All these data will proved to be a useful basis for study of glioma in vivo.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Brain Neoplasms ; pathology ; Disease Models, Animal ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Glioma ; pathology ; Magnetic Resonance Imaging ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Cells, Cultured

OBJECTIVETo study whether intraspinally transplanted human cord blood CD34+ cells can survive, differentiate, and improve neurological functional recovery after spinal cord injury in rats.

METHODSRats were randomly divided into two groups. One group of rats was subjected to spinal cord left-hemisection and transplanted with human cord blood CD34+ cells labeled by bromodeoxyuridine (BrdU); The other group was carried by left-hemisection with injection of PBS (control group). The neurological function was determined before and 24 h, 1, 2, 3 and 4 weeks after spinal cord injury and cell transplantation using the modified Tarlov score. The distribution and differentiation of transplanted human cord blood cells in vivo in rat spinal cord were evaluated by histological and immnuhistochemical analysis.

RESULTSFunctional recovery determined by modified Tarlov score was significantly improved in the group receiving human cord blood CD34+ cells compared with the control group (P < 0.05). Moreover, human cord blood CD34+ cells were found to survive in rat spinal cord microenvironment, with the expression of the neural nuclear specific protein (NeuN) in 2% BrdU-reactive human cells and of the astrocytic specific protein glial fibrillary acidic protein (GFAP) in 7% BrdU-reactive human cells.

CONCLUSIONSIntraspinally administered human cord blood CD34+ cells can survive, differentiate, and improve functional recovery after spinal cord injury in rats. Transplantation of human cord blood cells may provide a novel strategy for the treatment of neural injury.

4-Hydroxycoumarins ; Animals ; Antigens, CD34 ; metabolism ; Female ; Fetal Blood ; cytology ; Humans ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Recovery of Function ; Spinal Cord Injuries ; surgery ; Stem Cell Transplantation

OBJECTIVETo explore the regulational effect of oxidized low-density lipoprotein (Ox-LDL) on expression of type A scavenger receptor (SR-A) in human mesangial cells (HMC).

METHODSHMC line (HMCL) with high expression of SR-A (HMCL-SRA) was established after stable transfection of expressive vector with cDNA encoding SR-A. Uptake of Ox-LDL by HMCL was evaluated using Oil Red "O" staining. SR-A mRNA expression was examined using reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSMore uptake of Ox-LDL was observed in the HMCL-SRA than that in the untransfected HMCL. Ox-LDL could induce SR-A mRNA expression in HMC in a dose-dependent manner, and reached a peak level after 24 h of stimulation. After 24 h of stimulation with Ox-LDL at the dose of 10, 50 and 100 micrograms/ml, SR-A mRNA expression was up-regulated to 1.35, 1.83 and 2.30-fold of controls, respectively. However, LDL had no effect on the expression of SR-A.

CONCLUSIONSIt suggests that SR-A be a major binding receptor to uptake Ox-LDL in HMC. Ox-LDL may promote the progression of chronic renal diseases through up-regulation of SR-A.

Cells, Cultured ; DNA, Complementary ; Glomerular Mesangium ; cytology ; metabolism ; Humans ; Lipoproteins, LDL ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Immunologic ; biosynthesis ; genetics ; Receptors, Scavenger ; Reverse Transcriptase Polymerase Chain Reaction ; Scavenger Receptors, Class A ; Transfection ; Up-Regulation

OBJECTIVETo observe the effects of different human parathyroid hormone 1-34 (hPTH1-34) administration on SaoS-2 cells, and explore the mechanism of bone formation improvement.

METHODSEach cycle covered 48 h. SaoS-2 cells were continuously or intermittently stimulated by 50 ng/ml hPTH1-34 for 1, 3, 6, 12, and 24 h in each cycle. Total RNA was extracted by Trizol kit. Alkaline phosphatase (ALP), osteocalcin or bone Gla-containing protein (BGP) and cyclic adenosine monophosphate (cAMP) levels were measured by chemical method, radioimmunoassay and competitive protein binding method, respectively. c-fos gene expression was semi-quantified by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSALP level was time-dependently increased in 1, 3 and 6 h stimulation, especially in 3 and 6 h (compared with control, P < 0.01; P < 0.05 or P < 0.01 compared with continuous stimulation). The cAMP level was time-dependently increased in 3 and 6 h incubation (P < 0.05 compared with control and continuous stimulation). Intermittent hPTH1-34 stimulation had more effects on cAMP level than continous action (P < 0.001). hPTH1-34 intermittent stimulation of 1, 3, and 6 h enhanced c-fos gene expression time-dependently.

CONCLUSIONSIntermittent hPTH1-34 stimulation has a stronger effect on osteoblast than continuous action, especially in 3, 6 h in each cycle intermittent stimulation. The synchronous responses of c-fos, ALP and cAMP to hPTH1-34 suggest that hPTH1-34 affect Saos-2 cells through cAMP dependent protein kinase A (PKA) pathway and c-fos gene paly an important role.

Alkaline Phosphatase ; analysis ; Cells, Cultured ; Humans ; Osteoblasts ; cytology ; Osteocalcin ; analysis ; Osteogenesis ; drug effects ; Osteosarcoma ; genetics ; pathology ; Parathyroid Hormone ; pharmacology ; Parathyroid Hormone-Related Protein ; pharmacology ; Peptide Fragments ; pharmacology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo investigate the effects of long-term hormone replacement therapy (HRT) on the breasts of postmenopausal women using mammary ultrasonography.

METHODSAn open randomized clinical study was designed. The percutaneous estradiol gel was used in a cyclic regimen combined with micronized progesterone (MP) or medroxyprogesterone acetate (MPA). Sixty healthy women (natural menopause for 1 to 5 years) were recruited and divided into four groups according to the dosage of estrogen and two kinds of progestin. All were given for 25 days per month. Mammary ultrasonography was used to observe breast glandular section thickness, breast duct width, the morphology of lobular unit and the blood flow of color Doppler imaging at baseline and every year from the second to seventh year of HRT. The serum estradiol was also measured from the 15th to 25th day of the cycle. Breast pain was recorded by the subjects.

RESULTS(1) The breast glandular section thickness after HRT was larger than that of before HRT. The breast glandular section thickness became larger gradually over time while the breast duct width became smaller over time. The breast duct width of the fifth year of HRT was significantly different from that of the sixth year (P < 0.05). (2) Twenty-two persons had new breast structure changes after HRT, and the accumulated incidence was 41.5%. New solid lesions formation occurred in five subjects (8.3%) and new cyst formation occurred in one subject (1.7%). After the second year of HRT, the serum estradiol level of the subjects with breast structure changes was higher than that of without breast structure changes and in the sixth year of HRT, and the difference was significant (P < 0.05). After the second year of HRT, the breast glandular section thickness of the subjects with breast structure changes was larger than that of without breast structure changes and in the fifth and sixth year of HRT, the difference was significant (P < 0.05). (3) After HRT, the serum estradiol level of subjects with mastalgia was higher than that of without mastalgia and in the second and sixth follow-up year, the difference was significant (P < 0.05).

CONCLUSIONSThere is an increasing trend of the percentage of glandular tissues of the breast after HRT. There is an increasing trend of the serum estradiol level and the breast glandular section thickness among the subjects with the breast structure changes; there is an increasing trend of the serum estradiol level among the subjects with mastalgia. Mammary ultrasonography can be used to monitor breast structure changes and breast lesions during HRT.

Aged ; Breast ; pathology ; Estradiol ; therapeutic use ; Estrogen Replacement Therapy ; adverse effects ; Female ; Humans ; Medroxyprogesterone Acetate ; therapeutic use ; Menopause ; Middle Aged ; Time Factors ; Ultrasonography, Mammary

OBJECTIVETo investigate the structure and degradation property of the polyvinyl alcohol (PVA)-collagen complex drug membrane.

METHODSDrug collagen membrane was complexed with PVA. The physical and chemical properties of the membrane were characterized by transmission electron microscopy, scanning electron microscope, forier transform-infrared spectroscopy and differential scanning calorimetry. Degradation experiment was performed to determine the degradation property of membrane and a degradation curve was therefor drawn.

RESULTSThe thermodynamic stability of collagen membrane was not destroyed by adding PVA. Collagen had good compatibility with PVA. Compared with collagen membrane, collagen-PVA complex membrane had smaller and evener pores. Adding PVA decreased the degradation rate of membrane.

CONCLUSIONSPVA-collagen membrane has better microstructure and antidegradation property than collagen membrane.

Biocompatible Materials ; chemistry ; Collagen ; chemistry ; ultrastructure ; Humans ; Membranes ; Polyvinyl Alcohol ; chemistry ; Spectroscopy, Fourier Transform Infrared