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 .

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.

OBJECTIVETo study the feasibility of close and three-dimensional reconstruction of maxillary defects combination with pedicled buccal fat pad graft, temporalis myofascial flap and titanium mesh.

METHODSAmong 11 patients with maxillary tumors, 9 were malignant and 2 were benign. 8 cases underwent subtotal maxillectomy and 3 cases underwent total maxillectomy. Maxillary defects were closely reconstructed combination with pedicled buccal fat pad graft (BFP), temporalis myofascial flap (TMF) and titanium mesh to restore the profile of maxilla.

RESULTSAll flaps were alive with satisfactory function and aesthetic appearance. The uncovered BFP and the facial muscle surface of TMF epithelialized within 4-6 weeks. Their degree of mouth opening ranged from 2.5 to 4.0 cm and without nasal foods return. All patients had a good voice quality.

CONCLUSIONMaking use of BFP, TMF and titanium mesh can restore the good contour and function of maxillary. It is an effective method of reconstruction of maxillary defects, and is easy to manipulate. It is worthy to popularization.

Adipose Tissue ; Female ; Humans ; Male ; Maxilla ; Maxillary Neoplasms ; Middle Aged ; Prostheses and Implants ; Reconstructive Surgical Procedures ; Surgical Flaps ; Titanium

OBJECTIVETo investigate the clinical significance of erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) in the treatment of spinal tuberculosis.

METHODSSixty-seven patients (41 males and 26 females, ranging in age from 23 to 61 years) with active spinal tuberculosis in our hospital (from Mar. 2004 to Mar. 2007) were included in this study. The tuberculosis focus were located either in cervical spine, thoracic spine or in lumbar spine. After 4 to 6 weeks anti-tuberculosis chemotherapy, all the patients underwent one-stage operation (focus debridment) and auto-bone graft combined with internal fixation. Blood test for ESR and CRP were carried out at different times before and after operation.

RESULTSThe average ESR was (79.4 +/- 35.6) mm/h, and the average CRP was (44.3 +/- 17.5) mg/L before chemotherapy, indicating active tuberculosis focus. After 4 to 6 weeks chemotherapy, the average ESR was (45.3 +/- 21.0) mm/h,and the average CRP was (26.7 +/- 11.8) mg/L, the differences were statistically (P < 0.05), and the clinical symptoms of spinal tuberculosis relieved in all patients. Four weeks after operation, the average ESR dropped to (42.8 +/- 16.5)mm/h, the average CRP dropped to (23.8 +/- 10.0) mg/L statistically (P < 0.05). Eight weeks after operation, the average value of ESR and CRP were at normal level in 47 cases, indicating inactive tuberculosis focus. Focus healing was achieved in 65 patients after short-term chemotherapy.

CONCLUSIONThe level of ESR and CRP are high in active spinal tuberculosis and low when focus controlled. ESR and CRP are reliable parameters in evaluation the treatment and prognosis of spinal tuberculosis.

Adult ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Perioperative Care ; Tuberculosis, Spinal ; blood ; diagnosis ; drug therapy ; surgery ; Young Adult

1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.