Objective To investigate the mechanism of preventing islet β-cell apoptosis in NOD mice with pioglitazone.Methods Female NOD mice at 4 weeks of age were divided into pioglitazone group ( n =21,0.02％pioglitazone was added into the feed ) and control group ( n =21,fed with regular diet).The accumulative incidence of diabetes was followed-up to 52 weeks of age in each group of NOD mice.Pancreas was removed from NOD mice at 12 weeks of age in each group ( n =15 ) to score severity of insulitis by routine H-E staining.The apoptotic β-cells in islets were observed with double-labeling technique of TUNEL in situ combined with standard sensitive avidin-biotin complex (sABC) immunohistochemical method.The spleens were taken for cell culture; IL-4 and IFN-γ levels in sera and supernatants of cultured splenocyte,the activity of PPARγ and NF-κB nuclear proteins in cultured splenocyte were measured by ELISA.Results (1)At 30 and 52 weeks of age,the respective incidences of diabetes were 57.1％ and 76.2％ in pioglitazone group,and 76.2％ and 90.5％ in control group ( all P＞0.05 ).At 15 weeks of age,the incidence became 4.8％ in pioglitazone group,and 33.3 ％ in control group ( P =0.045 ).( 2 ) At 12 weeks of age,the percentages of non infiltrated islet and peri-insulitis islet in pioglitazone group were higher than those in control group ( 14.73％ vs 5.69％,P＜0.01 ; and 26.02％ vs 15.72％,P＜0.01 ),and that of intraislet insulitis was lower than that in control group ( 59.25％ vs 78.59％,P＜0.01 ).The percentage of apoptotic β-cell in pioglitazone group was lower than that in control group( 6.17％ ±3.62％ vs 10.62％ ±4.43％,P=0.008 ).(3) In sera,IFN-γ level in pioglitazone group was lower than that in control group [( 561.05±78.61 ) vs ( 666.43 ± 28.42 ) pg/ml,P =0.045].In cultured splenocyte supernatant,the level of IFN-γ in pioglitazone group was lower than that in control group[(605.84+65.60) vs (692.20+44.98) pg/ml,P=0.041].(4) In cultured splenocyte,PPARγ nuclear protein activity in pioglitazone group was higher than that in control group ( 0.06 ± 0.01 vs 0.03 ± 0.01,P =0.013 ),and NF-κB nuclear protein activity was lower than that in control group ( 0.03 ± 0.01 vs 0.08± 0.01,P =0.001 ).Conclusions Pioglitazone activates PPARγ nuclear protein,inhibits activity of NF-κB nuclear protein,downregulates IFN-γ,diminishes differeutiation of Th cells to Th1,and subsequently prevents insulitis and β-cell apoptosis in NOD mice.

Objective To investigate the mechanism of pioglitazone preventing diabetes and the role of nuclear factor of actived T cells (NFAT) on non-obese diabetic(NOD) mice .Methods (1)Female NOD mice at 4 weeks of age were randomly divided into pioglitazone group(n=21) and control group(n=21) .The accumulative diabetes incidence was followed-up to 30 weeks of age in each group of NOD mice .(2)Pancreas were removed from NOD mice at 12 weeks of age in each group(n=15) to score insulitis se-verity by routine HE staining .IL-4 ,IFN-γand peroxisome proliferator-activated receptor γ(PPARγ) mRNA levels in spleens were tested by RT-PCR .IL-4 and IFN-γlevels in sera ,the activity of PPARγand NFATc1 nuclear protein in spleens were measured by enzyme linked immunosorbent assay (ELISA) .Results (1) At 15 weeks of age ,the diabetes incidence was 4 .76% in pioglitazone group ,and 33 .33% in control group(P<0 .05) .At 30 weeks of age ,the diabetes incidence was 57 .14% in pioglitazone group ,and 76 .19% in control group(P>0 .05) .(2) At 12 weeks of age ,the insulitis score in pioglitazone group was lower than that in control group[(1 .79 ± 0 .75) vs .(2 .38 ± 0 .66) ,P<0 .05] .(3) IFN-γ mRNA level in pioglitazone group was lower than that in control group[(0 .16 ± 0 .07) vs .(0 .53 ± 0 .26) ,P<0 .05] ,and PPARγmRNA level in pioglitazone group was higher than that in control group(0 .91 vs .0 .25 ,P<0 .05) .(4)IFN-γ level in pioglitazone group was lower than that in control group [(561 .05 ± 78 .61)pg/mL vs .(666 .43 ± 28 .42)pg/mL ,P<0 .05] .(5)At 12 weeks of age ,the spleen PPARγnuclear protein activity in pioglitazone group was higher than that in control group [(0 .05 ± 0 .01) vs .(0 .02 ± 0 .01) ,P<0 .05)] ,and NFATc1 nuclear protein activity was low-er than that in control group[(0 .23 ± 0 .04) vs .(0 .33 ± 0 .04) ,P<0 .05] .Conclusion Pioglitazone could activate PPARγ nuclear protein ,inhibit activity of NFATc1 nuclear protein ,downregulate IFN-γ,diminish Th cells deviating to Th1 ,and sequently prevents insulitis and diabetes onset in NOD mice .

Objective To investigate the mechanism of Bushentongluoshugan Formula's effect in penile erection.Method All 40 adult male rats were randomly divided into the control group, castrated group, Bushentongluoshugan Formula group and sterandryl group.The rats of all groups, except the control group, were ablated bilateral testes. Bushentongluoshugan Formula group and sterandryl group were given the formula and sterandryl respectively. After 3 weeks the penile tissue was collected and the expression of transforming growth factor-β-mRNA (TGF-β-mRNA) was detected by using polymerase chain reaction (PCR) technique.Result The expression of TGF-β-mRNA in the castrated group was higher than that in other three groups, and among them the incremental order was the control group, sterandryl group and Bushentongluoshugan Formula group according to the decreasing performance of TGF-β-mRNA expression. The internal control of TGF-β-mRNA was less in the control group, sterandryl group and Bushentongluoshugan Formula group with a significant difference compared with that in the castrated group (P<0.01), and there was no significant difference between the sterandryl group and Bushentongluoshugan Formula group (P>0.05).Conclusion Bushentongluoshugan Formula plays an important role in penile erection through regulating the expression of TGF-β-mRNA.

OBJECTIVETo screen and characterize the short peptides which bind specifically to interleukin-2 (IL-2) receptor alpha chain (IL-2Ralpha) for acquisition of small antagonists for blocking the binding of IL-2 with IL-2Ralpha.

METHODS12-mer phage displayed peptide library was screened with the target cells of MT-2 cells which expressed IL-2Ralpha at high levels. The binding phage clones were eluted by anti-IL-2Ralpha monoclonal antibody. After 3 rounds of screening, the positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, and the amino acid sequences of the positive clones were deduced from the DNA sequences.

RESULTSSeven positive clones were screened out of the 17 phage clones bound to MT-2 cells. The positive clone M15 could bind specifically to MT-2 cell and PHA-activated peripheral blood monouclear cells. Amino acid sequence analysis identified 6 sequences, all of which contained hydrophilic residues, and 5 of these 6 sequences included Tyr, Phe and Leu conservative residues.

CONCLUSIONThe peptide sequences containing Tyr, Phe conservative residues identified in this study can bind to cell surface IL-2Ralpha.

Amino Acid Sequence ; Cell Line, Transformed ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Peptide Library ; Peptides ; genetics ; metabolism ; Protein Binding ; Receptors, Cell Surface ; metabolism ; T-Lymphocytes ; cytology ; metabolism

Adolescent ; Child ; Child, Preschool ; Clubfoot ; physiopathology ; Electromyography ; Female ; Humans ; Infant ; Male

Objective To explore the risk factors related to the formation of myocardial fatty infiltration and possible pathological consequences.Methods The macroscopic and microscopic findings in 117 autopsy cases with myocardial fatty infiltration were examined during October,2001 to June,2009.Results There was a significant positive correlation between the macroscopic grading of subepicardial adipose tissue and the microscopic myocardial fatty infiltrative degree(r_s=0.57,P ＜0.01)but there was no correlations between the myocardial fatty infiltrative degree and age as well as coronary arteriosclerosis (all P ＞0.05).The percent of myocardial atrophy was 39.32% (46/117),and the rate of myocardial atrophy in mild myocardial fatty infiltration group (13/63,20.63%) was significantly lower than that in moderate myocardial fatty infiltration group(22/39,34.92%;χ~2= 12.14,P ＜0.01) and in severe myocardial fatty infiltration group(11/15,73.33%;χ~2 = 13.42 ,P ＜0.01).There were 28 sudden cardiac deaths among the 117 eases including 6 deaths due to myocardial fatty infiltration.Conclusions Myocardial fatty infiltration is often associated with myocardial atrophy,even with sudden cardiac death but is not an accompanying pathologic changes of aging and coronary arteriosclerosis.

OBJECTIVETo study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).

METHODSWe established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.

RESULTSThe prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.

CONCLUSIONThe ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.

Animals ; Apomorphine ; Carcinogens ; Diethylstilbestrol ; Erectile Dysfunction ; etiology ; Humans ; Male ; Myocytes, Smooth Muscle ; ultrastructure ; Nitric Oxide Synthase Type I ; metabolism ; Organ Size ; Penile Erection ; Penis ; enzymology ; ultrastructure ; Pituitary Neoplasms ; chemically induced ; complications ; Prolactin ; blood ; Prolactinoma ; chemically induced ; complications ; Rats ; Rats, Wistar ; Testosterone ; blood

BACKGROUND: Polymethylmethacrylate (PMMA), commonly known as bone cement, has been widely used in the orthopedic surgery. It ensures the immediate stability of prosthesis and the minimal micromotion at the cement-bone interface, allowing early weight-bearing after surgery. OBJECTIVE: To investigate the biomechanical performance of Sanders type III fracture of the calcaneus by using PMMA bone cement as a treatment. METHODS: Eight adult cadaveric ankle and calcaneus specimens were selected and served as normal controls after detection of biomechanical properties. Another eight specimens were collected and randomized into experimental group and control group to make a model of Sanders type III fracture in the calcaneus. In the experimental group, PMMA bone cement was injected into the defect area. In the control group, the artificial bone was implanted in the defect area and a steel plate was used to fix the lateral calcaneus. Biomechanical properties of the specimens in the experimental and control group were detected. RESULTS AND CONCLUSION: (1) Strain and stress of the calcaneus: The stress distribution of the calcaneus in the normal control group was consistent with that in the experimental group, and there was no significant difference between the two groups. The stress of the calcaneus in the experimental group was similar to that in the control group with no significant difference. (2) Displacement and axial stiffness of the calcaneus: Compared with the normal control group, the calcaneal displacement in the experimental group only decreased slightly, and there was no significant difference between the two groups, and likewise, the calcaneal displacement in the control group increased slightly. In the experimental group, the axial compression strength was (21.98±1.88) MPa and the axial compression stiffness was (1 633±150) N/mm. Therefore, there was no significant difference between the experimental group and the normal control group (P > 0.05). (3) Contact strength of the subtalar joint: Fractures basically recovered with good outcomes after PMMA bone cement injection. To conclude, by using PMMA bone cement in the treatment of calcaneus fractures, the scientific validity and clinical utility can be ensured.

To investigate the interaction between tumor necrosis factor alpha (TNF alpha) mimotopes and TNF alpha-binding peptides screened from random phage display peptide library with TNF alpha mimotopes displayed on phage clone as the target, the computational docking program AutoDock (with confirmation calculations using Discover) was used to predict and analyze the binding modes of LLT-18 (TNF alpha binding peptide, sequence EHMALTYPFRPP) with TNF alpha, after which LCS-7 (TNF alpha mimic phage clone, displayed positive sequence c-RRPAQSG-c) was docked to LLT-18 manually. The binding between LLT-18 and TNF alpha or LCS-7 was stabilized predominantly through electrostatic interaction and H-bond formation. The Arg residues in TNF alpha or LCS-7 were important for their interaction with LLT-18. For LLT-18, the key amino acid residues were Glu1, His2, Met3 and Tyr7. These results suggest the feasibility of screening ligand to single epitope with specific phage clone as the target, and of predicting the interaction between small peptides by computer-aided molecular modeling.
Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; immunology ; Biotinylation ; Computer Simulation ; Drug Evaluation, Preclinical ; methods ; Epitopes ; immunology ; Humans ; Ligands ; Models, Molecular ; Oligopeptides ; chemistry ; metabolism ; Peptide Library ; Protein Conformation ; Solubility ; Tumor Necrosis Factor-alpha ; chemistry ; immunology ; metabolism

In this paper, we introduce the grid technology into the design of the distributed PACS. First, we analyse the architecture and functions of OGSA-DAI, and then, introduce a model of the distributed PACS based on OGSA-DAI, and give a detailed analysis on its basic components and workflow. Finally, we make a conclusion on the distributed PACS based on grid technology.
Computer Communication Networks ; Computer Systems ; Database Management Systems ; Humans ; Information Storage and Retrieval ; methods ; Medical Informatics ; Radiology Information Systems ; Software ; Software Design ; Systems Integration