Objective To observe the expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblast,and to study the role of fibronectin in pathogenesis of chronic fluorosis.Methods Male and female Wistar rats 144 were randomly divided into four groups,which were designated as the control group(normal diets,n =36),fluoride group(normal diets + 100 mg/L fluoride,n =36),lower calcium monophagia group (synthetic diets,n =36) and lower calcium monOphagia with fluoride group(synthetic diets + 100 mg/L fluoride,n =36).Rats were sacrificed 4 and 8 months after beginning of the experiment,respectively,and femur tissue was fixated and paraffin-embedded.The osteoblast isolated from calvaria of neonatal rats was treated with different dose of fluoride(0,1,2,4 mg/L fluoride,respectively) for 48 and 72 h,cell culture supernatant and cells were collected,respectively.The cranial osteoblasts were cultured in vitro and divided into four groups according to different concentration of fluoride added,which were 0(control group),0.01,1.00,and 10.00 mg/L groups.These cells were treated with mineralized induced medium at day 2 and cultured for 3 more weeks whereafter,and then the slides were fixed in alcohol.The expression of fibronectin in rat femur tissue was detected by immunohistochemistry (IHC),and fibronectin mRNA expression was determined by in situ hybridization; the fibronectin levels in supernatant of cultured osteoblast was examined by enzyme-linked immunosorbent assay(ELISA),and the expression of fibronectin mRNA in osteoblasts was detected with RT-PCR; skull mineralized nodule formation of osteoblasts was observed under a light microscopy after stained with 0.1％ red alizarin liquid.Results Little expression of fibronectin (brown granules under light microscope) could be seen in femur tissue of fluorosis rats of control group and lower calcium monophagia group; but abundantly expressed in fluoride group and lower calcium monophagia with fluoride group; the fibronectin was also expressed in osteoblasts,bone cells and bone marrow cells with less red particles in the control group and lower calcium monophagia group,but more in the fluoride group and lower calcium monophagia with fluoride group.The expression of fibronectin protein in supernatant of cultured osteoblasts was significantly increased in the group of 4 mg/L fluoride at 48 h(0.108 ± 0.042,t =0.764,P＜ 0.05) compared with control group(0.081 ± 0.010); the value was also significantly increased in 1,2,4 mg/L groups at 72 h(0.089 ± 0.010,0.087 ± 0.012,0.098 ± 0.023; t =0.765,0.704,0.996; all P ＜ 0.05) compared with control group (0.070 ± 0.014) ; the expression of fibronectin mRNA was much higher in 1,2,4 mg/L groups at 48 h (0.61 ±0.06,0.77 ± 0.07,0.77 ± 0.07) and 72 h(1.61 ± 0.14,2.54 ± 0.20,2.75 ± 0.22) compared with control group [0.48 ± 0.04(t =0.111,0.182,0.182,all P ＜ 0.05),0.97 ± 0.08(t =0.093,0.109,0.108,all P＜ 0.05) ].A lot of mineralized nodules could be seen under light microscope in 1.00 and 10.00 mg/L groups.Conclusions The expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblasts are increased,and fluoride also promotes the mineralization nodules formation of osteoblasts.These results suggest that fibronectin may regulate the process of bone mineralization,and possibly play a role in the development of skeletal fluorosis.

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

Objective To observe the time-dependent changes of astrocytes and microvessels in the ischemic core and surrounding areas.Methods Glial fibrillary acidic protein (GFAP)was measured by HE,and CD31 by immunohistochemistry as markers in post-mortem specimens from ten patients,who died of cerebral infarction.Results Each section of the infarcted brains was divided into four areas (the area 0-3).GFAP expressed a little in area 0 and 1,increased in a time-dependent manner in area 2 and 3;CD31 die not expressed in area 0,expressed a little in the area 1,and increased in area 2 and 3 continuously.Conclusions The proliferation of astrocytes and microvessels may play a significant role in the process of restoration after cerebral infarction.

Animals ; Bridged Bicyclo Compounds ; metabolism ; Cytochrome P-450 CYP3A ; biosynthesis ; genetics ; Drug Design ; Drug Interactions ; Enzyme Induction ; Humans ; Lithocholic Acid ; metabolism ; Phloroglucinol ; analogs & derivatives ; metabolism ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; physiology ; Receptors, Steroid ; genetics ; physiology ; Signal Transduction ; Terpenes ; metabolism ; Transcription Factors ; genetics ; metabolism

OBJECTIVETo study the rehabilitation effects ergometry on COPD patients.

METHODSThirty COPD out-patients in our Hospital were randomly divided into 2 groups. Rehabilitation group, 15 patients, performed leg ergometry exercise of 80% peak Watt x 30min/d x 3d/w x 12w. Another 15 patients were control group without exercise. All patients received conventional therapy. Pulmonary function testing (PFT), cardiopulmonary exercise testing (CPET), arterial blood gas analysis (ABG), Borg and CAT sores were done at both baseline and 12 w.

RESULTSThere was no statistically difference in lung function testing, blood gas analysis and cardiopulmonary exercise test when pre- exercises between 2 sub-groups. The IC, peak VO2 and peak, W of rehabilitation group significantly increased (P < 0.05); and Borg and CAT.scores significantly decreased (P < 0.05) from baseline; and other PFT and ABG did not change (P > 0.05). While there was no difference in control group (P > 0.05).

CONCLUSIONLeg submaximal ergometry rehabilitation improves health condition and ameliorate dyspnea symptoms in COPD patients.

Blood Gas Analysis ; Dyspnea ; therapy ; Exercise Test ; Exercise Therapy ; Humans ; Pulmonary Disease, Chronic Obstructive ; therapy ; Respiratory Function Tests

OBJECTIVEThe expression of C/EBPalpha protein and mRNA during automatically activation process in primary cultures of HSCs were observed in order to explore its possible association with the proliferation and activation of HSCs.

METHODSImmunocytochemistry, Western blot and RT-PCR were used to evaluated the expression of C/EBPalpha protein and mRNA; as well as the expression of alpha-SMA, Desmin, MMP2, type I procollagen (alpha1). The eukaryotic vector harboring the full length cDNA of C/EBPalpha was transfected into activated HSC, then immunocytochemistry was applied to confirm the transfection and evaluate the effect of transfection on the proliferation of HSC by calculating the PCNA-positive cells. The morphological changes of HSC were observed by use of phase-contrast microscope.

RESULTSConstitutive expression of mRNA and protein of C/EBPalpha were detected in primarily cultured HSCs, and the protein was seen in both nuclei and cytoplasm with the latter being dominant. Their expression levels reached highest at day 2 of the culture, then decreased gradually when continually cultured to the day 4, 7, 10, on the other hand, the expression of alpha-SMA, MMP2 and ColI(alpha1) increased steadily. Transient transfection was verified by the fact that much more and stronger C/EBPalpha stain was observed in transfected HSCs than in void-vector transfected cells. In C/EBPalpha gene transfected HSCs, the number of PCNA-positive cells dramatically decreased compared with the void-vector transfected cells 24h after transfection. In addition, the C/EBPalpha gene transfected HSCs died 36 h after transfection, a few surviving cells became longer and thinner in morphology, however the void-vector transfected cells almost all remained alive.

CONCLUSIONSC/EBPalpha was likely involved in the HSCs activation, and over-expressed C/EBPalpha by transfection had inhibitory influence on the proliferation of cultured rat HSCs.

Animals ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; Cells, Cultured ; Collagen Type I ; genetics ; Liver ; cytology ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo study the effect of RAR-beta transfection plus treatment with the corresponding ligand ATRA on the proliferation and phenotype of platelet-derived growth factor (PDGF)-activated hepatic stellate cells (HSC).

METHODSPDGF-activated hepatic stellate cells of rats were transfected with eukaryotic expression vector pCMV-script-RAR-beta, which was verified by western blot. The proliferation of transfected HSC was assayed by BrdU incorporation as well as MTT methods. Their phenotype (alpha-SMA and desmin) was observed by immunocytochemistry assay with image analysis and RAR-beta protein expression was detected by western blot.

RESULTSTransfection of RAR-beta gene and treatment with ligand ATRA could increase the expression of RAR-beta protein for at least 144h and inhibit the proliferation and the expression of alpha-SMA and desmin in PDGF-activated HSC. Significant statistical differences were perceived comparing with sham-transfected, only-PDGF treated, non-ligand treated and irrelevant ligand-treated HSC.

CONCLUSIONSTransfected with RAR-beta gene as well as using related ligand ATRA could suppress the proliferation and reverse the activation phenotype of activated HSC.

Animals ; Blotting, Western ; Cell Division ; Liver ; cytology ; Phenotype ; Platelet-Derived Growth Factor ; pharmacology ; Rats ; Receptors, Retinoic Acid ; physiology ; Transfection ; Tretinoin ; pharmacology

OBJECTIVETo establish a method for determining anticoagulation potency of Sparganii Rhizoma, and evaluate the effect of Sparganii Rhizoma herbs from different producing areas on promoting blood circulation and removing blood stasis; and study the material basis of Sparganii Rhizoma through the correlation analysis on its anticoagulation potency, ferulic acid and total flavonoid content.

METHODThe anticoagulation time of Sparganii Rhizoma from different producing areas with activeated partial thromboplastin time for their active extracts. Their biopotency was calculated by using the method of "parallel lines of dose effect" (3, 3). The degree of correlation between their anticoagulation potency and chemical constituents were calculated by using Pearson correlational analysis method.

RESULTSparganii Rhizoma and is control drugs had a good linear relationship between dose and effect (Y = 172.76X - 193.39, R2 = 0.9955). The method had better accuracy (RSD 4.7%), repeatability (RSD 2.3%) and intermediate precision (RSD 5.4%), finding that the biopotency of Sparganii Rhizoma from different producing areas ranged between 52.33-238.58 U x g(-1), and all of them passed the test on reliability. The results of correlation analysis showed no remarkable relationship between the anticoagulation potency of Sparganii Rhizoma and the contents of the two chemical constituents.

CONCLUSIONThis biopotency determination method established in the experiment can be used as one of approaches for qulaity evaluation on Sparganii Rhizoma.

Animals ; Anticoagulants ; chemistry ; pharmacology ; Blood Coagulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Rabbits ; Rhizome ; chemistry ; Typhaceae ; chemistry

OBJECTIVETo assess the nutrition status of children and adolescents in China using the WHO growth reference (2007) in comparison with that defined by the International Obesity Task Force (IOTF) and the Working Group on Obesity in China (WGOC).

METHODSOverweight and obesity were defined by age-, sex-, specific BMI reference developed by WHO (2007), IOTF (2000), and WGOC (2004), respectively. Stunting and thinness were defined as height and BMI less than two standard deviations (SD) of the WHO growth reference (2007), respectively. Data of children and adolescents aged 5 to 19 years (n=54,857, 28,273 boys, 26,584 girls) from the 2002 China National Nutrition and Health Survey (CNNHS) were used in the study.

RESULTSThe prevalence of overweight, obesity, stunting and thinness among Chinese children and adolescents aged 5-19 years was 5.0%, 1.2%, 13.8%, and 7.4%, respectively when the WHO growth reference (2007) was used, whereas the estimated absolute total number affected by these 4 conditions were 14.6, 3.7, 40.6, and 21.8 million, respectively. The prevalence of overweight and obesity was 18.1% in large cities, while the stunting prevalence was 25.1% in rural 4. Obesity prevalence assessed by the WHO growth reference was higher than that as assessed by the IOTF reference, and obesity prevalence assessed by the WGOC reference was lower than that as assessed by the IOTF reference.

CONCLUSIONThe nutritional status of children and adolescents is not equal in different areas of China. Stunting is still the main health problem of the poor, while overweight and obesity are the main health problems in large cities.

Adolescent ; Adult ; Age Distribution ; Body Weight ; Child ; Child Nutrition Disorders ; epidemiology ; Child Nutritional Physiological Phenomena ; Child, Preschool ; China ; Female ; Humans ; Male ; Nutrition Surveys ; Nutritional Status ; Sex Characteristics ; World Health Organization ; Young Adult

OBJECTIVEAntigen-presenting cells such as monocytes and dendritic cells (DCs) stimulate T-cell proliferation and activation during adaptive immunity. This cellular interaction plays a role in the growth of atherosclerotic plaques. Tanshinone II A (TSN) had been shown to decrease the growth of atherosclerotic lesions. We therefore investigated the ability of TSN to inhibit human monocyte-derived DCs and their T-cellstimulatory capacity.

METHODSDCs derived from human monocytes cultured with recombinant human interleukin (IL)-4 and recombinant human granulocyte-macrophage colony-stimulating factor were co-cultured with TSN and lipopolysaccharide for 48 h. Phosphate-buffered saline was used as a negative control. Activation markers and the capacity of DCs for endocytosis were measured by flow cytometry, and proinflammatory cytokines were measured by enzyme-linked immunosorbent assays. DCs were co-cultured with lymphocytes to measure T-cell proliferation and IL-2 secretion by mixed lymphocyte reactions.

RESULTSTSN dose-dependently attenuated DC expression of costimulatory molecules (CD86), and decreased expression of major histocompatibility complex class II (human loukocyte antigen-DR) and adhesion molecules (CD54). Moreover, TSN reduced secretion of the proinflammatory cytokines IL-12 and IL-1 by human DCs, and restored the capacity for endocytosis. Finally, TSN-preincubated DCs showed a reduced capacity to stimulate T-cell proliferation and cytokine secretion.

CONCLUSIONSTSN inhibits DC maturation and decreases the expression of proinflammatory cytokines, while impairing their capacity to stimulate T-cell proliferation and cytokine secretion. These effects may contribute to the influence of TSN on the progression of atherosclerotic lesions.

Antigen-Presenting Cells ; drug effects ; Atherosclerosis ; immunology ; pathology ; B7-2 Antigen ; metabolism ; Cell Membrane ; drug effects ; metabolism ; Cytokines ; secretion ; Dendritic Cells ; drug effects ; immunology ; secretion ; Diterpenes, Abietane ; pharmacology ; Endocytosis ; drug effects ; Flow Cytometry ; Humans ; Immunity, Cellular ; drug effects ; Inflammation Mediators ; metabolism ; Lymphocyte Activation ; drug effects