Objective To explore the relationship between the expression level of p27 KIP1 and p57 KIP2 and the development and prognosis of thyroid carcinoma. Methods p27 KIP1 and p57 KIP2 expression in 6 nodular goitres, 10 thyroid adenomas and 35 thyroid carcinoma were detected by immunohistochemical method, and the relationship between p27 KIP1 and p57 KIP2 expression and clinicopathological characteristics were analyzed. Results The positive rate of nuclear expression of p27 KIP1 and p57 KIP2 in nodular goitres and thyroid adenomas was obviously higher than that in thyroid carcinomas, in papillary carcinoma and follicular carcinoma was significantly higher than that in medullary carcinoma and undifferentiated carcinoma, and in the thyroid carcinomas of stages I and II was also significantly higher that in the thyroid carcinomas of stages III and IV. The frequency of cytoplasmic expression of both p27 KIP1 and p57 KIP2 in thyroid carcinoma with lymph node metastasis was obviously higher than that in ones without lymph node metastasis. Conclusion The nuclear expression of both p27 KIP1 and p57 KIP2 in thyroid carcinoma were significantly associated with clinical stage, lymph node metastasis and local invasion. The low nuclear expression of p27 KIP1 and p57 KIP2 may indicate the poor prognosis of the patients, and the cytoplasmic expression may indicate the lymph node metastasis.

Objective:To determine the plasma level of apelin in patients of maintenance hemodialysis (MHD) and to explore the relationship between apelin level and carotid atherosclerosis (AS) in MHD patients.Methods:A total of 92 MHD patients and 36 sex-and age-matched healthy subjects were enrolled in this study.The plasma level of apelin was evaluated by radiation immunoassay;serum endothelial injury markers including thrombomodulin,von Willebrand factor (vWF),and CD 146,and inflammatory factors including high sensitive C-reactive protein (hsCRP),IL-6 and TNF-α were determined by ELISA.Common Carotid arteries intima media thickness (CCA-IMT),cross-sectional calculated intima-media area (cIM area) area and atherosclerotic plaque were measured by non-invasive high-resolution B-mode ultrasonography.Results:The plasma levels of apelin was significantly decreased in MHD patients compared with healthy subjects (P＜0.01),accompanied with elevated plasma levels of thrombomodulin,vWF,CD 146,hsCRP,IL-6 and TNF-α (all P＜0.01).The plasma levels of apelinin in MHD patients with carotid artery plaques were obviously lower than those without plaques [(43.16± 10.12) pg/mL vs (61.43±16.25) pg/mL,P＜0.01].Plasma level of apelin was inversely related with CCA-IMT and cIM area (r=-0.355 and r=-0.297 respectively,all P＜0.01).Multiple stepwise regression analysis showed that plasma level of apelin was an independent risk factor for CCA-IMT and cIM area.Conclusion:The plasma apelin in MHD patients might take part in vascular endothelial injury and the progress of atherosclerosis.It plays an important role in the initiation and development of uremia associated atherosclerosis through elevating inflammatory factors including hsCRP,IL-6 and TNF-α levels.

Background Many researches confirmed that basic fibroblast growth factor (bFGF)plays an important role on the proliferation and differentiation of retinal progenitor cells,but its low intraocular permeability limits its clinical application.To explore an effective approach to enhance the intraocular permeability of bFGF has an important significance for the treatment of retinopathy. Objective This study was to investigate the effect of azone on bFGF intraocular permeability after its topical administration. Methods Eighteen New Zealand white rabbits were randomly divided into four groups on random number table method.Distilled water( blank control group),5％ bFGF eyedrops(5％ bFGF group ),0.4％ azone+5％ bFGF eyedrops (0.4％ azone + 5％ bFGF group ) and 0.4％ azone+ 10％ bFGF eyedrops (0.4％ azone + 10％ bFGF group)were topically administered in different groups at 5- minute interval for 3 times.Aqueous and vitreous fluid were extracted 30 minutes after administration of eyedrops,and those in the 0.4％ azone + 5％ bFGF group were obtained 30,60 and 120 minutes after administration.bFGF concentration in the aqueous and vitreous fluid was quantified with ELISA. Results The bFGF levels(A value)in aqueous and vitreous fluid were 0.1007±0.0100 and 0.1340±0.0100 after topical administration of the 5％ bFGF eyedrops,those in blank control group were 0.1363 ±0.0100 and 0.1130±0.0100,respectively,and those in the 0.4％ azone+5％ bFGF group and 0.4％ azone+10％ bFGF group were significantly higher than the 5％ bFGF group ( both P=0.000).However,no significant difference was found in bFGF levels between 0.4％ azone+5％ bFGF group and 0.4％ azone + 10％ bFGF groups in both aqueous and vitreous fluid ( P =0.985,0.098 ).A value of bFGF in aqueous was gradually increased with prolong of time in the 0.4％ azone+5％ bFGF group,with the values 0.9413±0.0300 at 30 minutes,0.3865±0.0300 at 60 minutes,and 0.2550±0.0300 at 120 minutes,showing a positive linear correlation between bFGF level and time( R2 =0.736,P =0.003 ),but no significant correlation was seen in vitreous sample(R2=0.196,P=0.233). Conclusions Azone can improve the intraocular penetration of bFGF eyedrops.Increasing the concentration of bFGF in eyedrop from 5％ to 10％ dose not change its intraocular distribution.The highest content of the bFGF in aqueous is at 30 minutes following the administration of 0.4％ azone+5％ bFGF eyedrops.

Objective To investigate the effects of diabetic mellitus on the opening of mycardial mitochondrial permeability transition pore (MPTP) in rats,and to explore the role of the opening of MPTP in the development of diabetic cardiomyopathy.Methods Sixteen male Wistar rats were randomly divided into normal control group (NC) and diabetic mellitus group (DM) (8 each).Diabetic mellitus model was reproduced by a single intraperitoneal injection of streptozotocin (STZ).After 12 weeks,the production rate of mitochondrial reactive oxygen species (ROS) and the change in mitochondrial membrane potential (MMP) in myocardial mitochondria were assayed by fluorospectrophotometer.Cardiomyocyte apoptosis was evaluated by terminal-deoxynucleotidyl transferase mediated deoxy-UTP nick end labeling (TUNEL),and the glutathione (GSH) level in serum was measured by spectrophotometer.Results Compared with NC group [13.01?3.31U/(s?mg)],the production rate of mitochondrial ROS was significantly increased in DM group [28.79?6.11U/(s?mg),P

Objective To investigate the effect of telmisartan on mitochondrial membrane potential and cardiomyocyte apoptosis of rats with type 1 diabetes.Methods Type 1 diabetic rat model was reproduced by intraperitoneal injection of streptozotocin to adult male Wistar rats.Sixteen diabetic rats were randomly divided into two groups (8 each):DM group and T group (rats were treated with telmisartan).Another 8 normal rats served as control (Con group).After 12 weeks,the levels of reactive oxygen species (ROS) were detected by fluorescent probe DCFH-DA,and the changes in mitochondrial membrane potential (?m) was detected by fluorescent probe JC-1.Cardiomyocyte apoptosis was evaluated by TUNEL.Results The levels of blood glucose were higher and body weight was lower in both DM and T group than in Con group at 7th day and 12th week after streptozotocin injection (P

Objective To explore the feasible approach for establishment of a liver cancer model induced by N-nitrosodiethylamine(DENA)with Sprague-Dawley rat,and to provide ideal animal model for imaging diagnosis and interventional therapy.Methods One-hundred and forty male SD rats were administrated with 0.95 g/L DENA for 10 weeks,and MRI was performed for inspecting pathological changes of rat livers on the following week.When the liver tumor was proved then the rat will be the candidate for sequential procedures,otherwise the animal continued under observation until next MRI examination after 4 weeks.DSA was done in 64 rats'for detecting blood supply of liver tumors.Animals were sacrificed with an overdose of chloral hydrate,The representative tumor tissues were fixed in 10% formalin and 2.5% glutaraldehyde for light and electron microscopy analysis respectively.Alpha fetoprotein(AFP) and hepatocyte were assaied by immunohistochemistry technique in order to identify intrinsic trait of the harvested tumors.Results The earliest induced tumor was detected on 11th week and the latest was on 20th week by MRI,and the median period was 13.9 weeks.Tumor size ranged from 2 mm to 40 mm in diameter. The rate of single and multi-induced tumor was 9.7%(7/72)and 90.3%(65/72),respectively.87.7% (57/65)" of the induced multiple tumors was with hepatocirrhosis and 18.1% of these tumors combined with extrahepatic neoplasm or metastasis.Plenty blood supply was proved by DSA in most of those tumors. Tumors not only derived from hepatocyte but also manifested positive expression of AFP.Histological types of these tumors include hepatocellular carcinoma(HCC)(92.0%,66/72),intrahepatic cholangiocarcinoma (ICC)(4.0%,3/72),and combined HCC and ICC(4.0%,3/72),respectively.Electro-microscope analysis indicated that cytoplasm and organelle of induced tumors were abnormal distinctly compared with those of non-carcinomatous cells.Conclusion DENA can induce ideal rat liver cancer as a feasible animal model,MRI is the best approach for scrutinizing pathological changes of rat livers during induced period.

Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.

Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.

ObjeclJve To definine the corresponding value to glycated albumin(GA)for a specific target of HbAlc,and to elvaluate the relationship between GA and HbA1c.Methods From Oct.2006 TO Apr.2009, 2 532 subjects were enrolled who accepted oral glucose tolerance test(OGtt)in out-patient department,including 898 with normal glucose regulation,695 with impaired glucose regulation,and 939 with newly-diagnosed diabetes.GA was measured with liquid enzymatic method.HbA1c was measured with high performance liquid chromatography method.The plasma glucose was measured at fasting,0.5 h,1 h,2 h,and 3h after glucose load.The correlation among GA,HbA1c and the other parameters monitored was analyzed.Results (1)The levels of HbA1c and GA in 2 532 subjects were(6.3±1.1)% and(17.9±4.5)%.The ratio of GA/HbA1c was 2.85±0.51.(2)HbAlc and GA were positively correlated with fasting,0.5 h,1 h,2 h and 3 h plasma glucose(r was in 0.567-0.776,atl P<0.01).(3)GA was significantly correlated with HbA1c(r=0.701,P<0.01).Linear regression analysis,using GA and HbA1c summarized by patient(n=2 532),produced a relationship of GA=2.871×HbA1c-0.112.The change in GA per increase of 1% HbA1c was 2.87%.When HbA1c level was 6.5%,the expected value of GA was 18.5%.The sensitivity,specificity,positive predictive value,negative predictive value,and accuracy with GA≤18.5% to predict HbA1c≤6.5% were 82.32%,72.49%,86.48%.65.73%,and 79.19%,respectively.When HbA1c level was 7.0%,the expected value of GA was 20.O%.When HbA1c level was 7.5%.the expected value of GA was 21.4%.Conclusions We initially establish the corresponding value to GA for a specific target of HbA1c and provide the basis for clinical application.

The purpose of the study was to observe the effect of rapamycin (RAPA) on the differentiation and maturation of rat bone marrow-derived dendritic cells (BMDCs) in vitro. BMDCs from Wistar rats were cultured with granulocyte-macrophage colony-stimulating factor plus interleukin-4 in the presence or absence of RAPA (20 ng/mL), and stimulated with lipopolysaccharide (LPS) for 24 h before cells and supernatants were collected. Surface phenotype of BMDCs was flow-cytometrically detected to determine the expression of maturation markers, MHC class II and CD86. Supernatants were analyzed for the production of IL-12 and IFN-gamma cytokines by using ELISA. BMDCs were co-cultured with T cells from Lewis rats and mixed lymphocyte reaction was assessed by MTT method. The morphology of BMDCs stimulated with LPS remained immature after RAPA pretreatment. RAPA significantly decreased the CD86 expression, impaired the IL-12 and IFN-gamma production of BMDCs stimulated with LPS, and inhibited the proliferation of allogeneic T cells. In conclusion, RAPA can inhibit the maturation of BMDCs stimulated with LPS in terms of the morphology, surface phenotype, cytokine production, and ability of BMDCs to stimulate the proliferation of allogeneic T cells in vitro.