Objective To investigate the effects of blockade of OX40/OX40L costimulation pathway on mice islet allograft tolerance in CD40/CD154 costimulation pathway blockade mice.Methods C57BL/6 mice were induced into diabetes mellitus as recipients,and were transplanted with DBA/2 mice islets.The recipients were divided into four groups,(1) treated with IgG as controls,(2) anti-OX40L mAb,(3) anti-CD154,(4) combined treatment of anti-OX40L mAb and anti CD154mAb.The mean survival time (MST) of islet allograft was observed.The expression of OX40 in activated T cells of CD154 deficient mice was detected.Effector T cells were obtained from the spleen of CD154 deficient mice cultured with or without anti-OX40L mAb for 3 days.The proliferation of T cells was assayed.Results The MST in the control group,anti-OX40L mAb group,anti-CD154 mAb group and anti OX40L mAb + anti-CD154 mAb group was 19,22,48,and ＞150 days respectively (P ＜0.05).The OX40 expression was readily induced in the 66％ activated T effector cells.CD154 deficient T effector cells proliferation was inhibited by the addition of anti-OX40L mAb in the culture in a dose-dependent fashion.Conclusion The blockade of OX40/OX40L costimulation pathway can promote islet allograft tolerance in CD40/CD154 costimulation pathway blockade mice by inhibiting the proliferation of T cells.

Purpose To analyze differential diagnosis between Hashimoto thyroiditis of atypical cell clusters ( ACC) and papillary thy-roid carcinoma (PTC). Methods 153 cases of Hashimoto thyroiditis (HT) were collected and divided into 3 groups:PTC group (49 cases), ACC group (32 cases), and HT as control group (72 cases). Morphology observations were done. CD56, CK19, galectin-3 and Ki-67 were detected by immunohistochemistry. Results Morphologic differences were observed among the groups. PTC showed milder positive expression of CK19 and galectin-3, and weaker positive expression of CD56 than that of ACC. Ki-67 index in ACC and PTC was lower than that of HT. Conclusions Morphological characteristics combined with CD56, CK19, Galectin-3 and Ki-67 evalu-ations could be valuable in the differential diagnosis between PTC and ACC.

OBJECTIVETo investigate the feasibility of dynamic magnetic resonance diffusion tensor imaging (DTI) in rabbit models with implanted VX2 hepatic tumor grafts.

METHODSMRI and DTI images were obtained from 16 rabbit models with implanted VX2 hepatic tumor grafts (14, 18, 22, and 26 days after tumor implantation, respectively) and 4 normal rabbits. The average diffusion coefficient (ADC) and exponential ADC (eADC) were estimated and compared against pathological findings.

RESULTSThe ADC values increased after tumor implantation but then decreased in the rabbit models, whereas eADC exhibited a pattern of reverse changes. These changes significantly differed from those in the control group. Coagulation necrosis and fibrous hyperplasia showed obvious increase as found by pathological examination.

CONCLUSIONDynamic MR DTI quantitative analysis of rabbit models of implanted VX2 hepatic tumor can partially describe the growth behaviors of implanted liver cancer.

Animals ; Carcinoma, Hepatocellular ; pathology ; Diffusion Magnetic Resonance Imaging ; Female ; Image Enhancement ; methods ; Liver Neoplasms ; pathology ; Male ; Neoplasm Transplantation ; Rabbits ; Random Allocation

The superantigen,such as staphylococcal enterotoxins, had been identified as possible anti-cancer molecules in many reports. In this paper, we cloned the entA gene encoding Staphylococcal enterotoxin A from the genomic DNA of Staphylococcus aureus(ATCC13565) by PCR, the sequence cloned was accordance with that reported in Genebank. The entA gene could be expressed effectively after inserted into plasmid pET-22b( + ), The rSEA was expressed as inclusion bodies when induced by IPTG at 37 degrees C and became soluble after induced at low temperature, the soluble part is about 55% of total rSEA products. Only one band was detected by western-blotting in expression product of BL-21 (DE3) with pET-SEA. The soluble rSEA was purified by Ni2+ chelating sepharose column. No other protein except rSEA was seen in SDS-PAGE gel stained by both Coomassie brilliant blue and silver salt, which showed that the rSEA was purified effectively. Homology modeling of rSEA determined the structure change was conducted, which indicated there was no apparent structure change between rSEA and native SEA. This result was also confirmed by proliferation assay of PBMC, for the rSEA could induced proliferation of PBMC as effectively as native SEA. The increasing anti-tumor activity of rSEA was also detected after the spleen cell activated in vivo by rSEA, which was accordance with others reports. This work paved the way for the further study of anti-cancer with rSEA.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Humans ; Leukocytes, Mononuclear ; cytology ; drug effects ; Plasmids ; Polymerase Chain Reaction ; Solubility

As the ubiquitous nucleic acids recognizing motif, Zinc finger protein play important role in regulation of gene expression. The study of recognization specific will greatly facilitate understanding the delicate interaction of Zinc finger protein and DNA. By the choice of expression vector, the induction and culture conditions, the DNA binding domain of Zif268 was expressed in Escherichia coli partly solubly. The gel mobility shift assay shows that purified DNA binding domain can bind its natural target sequence specifically, which indicates the DNA binding domain remains its DNA binding activity in Escherichia coli. The functional expression of DNA binding domain of Zif268 will greatly facilitate the development of in vivo genetic selection assay for the study of Zinc fingers-DNA interaction.
DNA-Binding Proteins ; genetics ; Electrophoretic Mobility Shift Assay ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; Zinc Fingers ; genetics

Objective To evaluate the effect of a novel orthopedic biomaterial,graded composite zireonia(ZrO2)hydroxyapatite(HAP)on activity of rat osteoblast ceils(OB)cultured in vitro. Methods The pure zirconia material was used as control to measure surface roughness of the composite material that was examined by using scanning electron microscope(SEM)and X-ray diffraetometer (XRD).The rat osteoblast cells were cultured on the two materials.Alkali phosphatase(ALP)of the two groups was measured and ELISA was used to detect IL-6 and TGF-?eoncentration of the supematant of OB cells.Tumor growth factor-?(TGF-?)mRNA was detected by RT-PCR.SEM was used to observe OB cells on the two materials.The extract of the composite material was used for a eytotoxicity test to cal- culate the relative proliferation rate(RGR)and classify the toxicity.Results The surface roughness of the gradual composite materials was significantly higher than that of the control materials(P＜0.01). The ALP of the gradual material group was markedly higher than that of the control group at different in- tervals.There was significant difference of the IL-6 and TGF-?concentrations 2-4 days after culture be- tween two groups(P＜0.05,P＜0.01).The mRNA level of TGF-?of the two OB groups also showed marked statistical difference(P＜0.01).The ossification of the OB cells on the composed material was marked after 14 days.The MTT color experiments showed no statistic significance between materials group and negative group,with the toxicity at levelⅠand 0(P＜0.05).Conclusion Graded composite ZrO2 HAP can significantly promote proliferation and differentiation of OB cells cultured in vitro and has good biocompatibility.

OBJECTIVETo determine differences in adherence to secondary prevention guidelines (pharmacological interventions) among coronary heart disease (CHD) patients between a Chinese medicine (CM) hospital and a general hospital in a Chinese city.

METHODSMedical records of 200 patients consecutively discharged from the CM hospital and the general hospital for CHD were reviewed to determine the proportions of eligible patients who received antiplatelet agents, β-blockers, angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs) and statins at discharge. The effects of patient characteristics and hospital type on the use of these medicines were estimated using logistic regression models.

RESULTSPatients discharged from the CM hospitals were older; more likely females; had greater history of hyperlipidemia, cerebrovascular diseases and less smoker (P<0.01 or P<0.05). They were less likely to receive coronary angiography and percutaneous coronary intervention, and had a longer length of stay than those discharged from the general hospital (P<0.01 or P<0.05). There were no significant differences in antiplatelet agents (96% vs. 100%, P=0.121) or statins (97.9% vs. 100%, P=0.149) use between the CM hospital and the general hospital. In multivariable analyses that adjusted for patient characteristics and hospital type, there was no significant difference in use of β-blockers between the CM hospital and the general hospital. In contrast, patients discharged from the CM hospital were less likely to receive ACE inhibitors/ARBs compared with those discharged from the general hospital (odds ratio: 0.3, 95% confidence interval: 0.105-0.854).

CONCLUSIONIn this study, the CM hospital provides the same quality of care in CHD for prescribing evidence-based medications at discharge compared with another general hospital except for ACE inhibitors/ARBs use.

Aged ; Coronary Disease ; drug therapy ; prevention & control ; Evidence-Based Medicine ; Female ; Hospitals, General ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Secondary Prevention

AIMStudies on synthesis and antibacterial activity of new heterocycles.

METHODSThe cyclocondensation of [(3-pyridyl)-1,3,4-oxadiazol-2-yl] thio acetic acid with various aroyl hydrazines in the presence of POCl3 and xylene gave the corresponding titled compounds, and the in vitro antibacterial activity was primarily evaluated by the method of cupplate diffusion solution.

RESULTSSixteen novel titled compounds were synthesized, their structures were confirmed by IR, 1HNMR, MS and elemental analysis. Biological screening results demonstrated that most of the compounds prepared displayed potential antibacterial activity.

CONCLUSIONOxadiazoles incorporting pyridyl oxadiazole ring may be usefully antibacterial candidate drugs.

Anti-Bacterial Agents ; chemical synthesis ; chemistry ; pharmacology ; Escherichia coli ; drug effects ; Oxadiazoles ; chemical synthesis ; chemistry ; pharmacology ; Proteus vulgaris ; drug effects ; Staphylococcus aureus ; drug effects

OBJECTIVE: To determine the relationship between intravoxel incoherent motion (IVIM) imaging derived quantitative metrics and serum soluble CD40 ligand (sCD40L) level in an embolic canine stroke model. MATERIALS AND METHODS: A middle cerebral artery occlusion model was established in 24 beagle dogs. Experimental dogs were divided into low- and high-sCD40L group according to serum sCD40L level at 4.5 hours after establishing the model. IVIM imaging was scanned at 4.5 hours after model establishment using 10 b values ranging from 0 to 900 s/mm². Quantitative metrics diffusion coefficient (D), pseudodiffusion coefficient (D*), and perfusion fraction (f) of ischemic lesions were calculated. Quantitative metrics of ischemic lesions were normalized by contralateral hemisphere using the following formula: normalized D = D(stroke) / D(contralateral). Differences in IVIM metrics between the low- and high-sCD40L groups were compared using t test. Pearson's correlation analyses were performed to determine the relationship between IVIM metrics and serum sCD40L level. RESULTS: The high-sCD40L group showed significantly lower f and normalized f values than the low-sCD40L group (f, p < 0.001; normalized f, p < 0.001). There was no significant difference in D*, normalized D*, D, or normalized D value between the two groups (All p > 0.05). Both f and normalized f values were negatively correlated with serum sCD40L level (f, r = −0.789, p < 0.001; normalized f, r = −0.823, p < 0.001). However, serum sCD40L level had no significant correlation with D*, normalized D*, D, or normalized D (All p > 0.05). CONCLUSION: The f value derived from IVIM imaging was negatively correlated with serum sCD40L level. f value might serve as a potential imaging biomarker to assess the formation of microvascular thrombosis in hyperacute period of ischemic stroke.
Animals ; CD40 Ligand* ; Diffusion ; Dogs ; Infarction, Middle Cerebral Artery ; Magnetic Resonance Imaging* ; Perfusion ; Stroke* ; Thrombosis

BACKGROUNDA new lacunar infarction model was recently established in beagle dogs through proximal middle cerebral artery (MCA) occlusion by thrombus. This study aimed to characterize the model by multimodal magnetic resonance imaging (MRI) and to investigate its potential role for the future stroke research.

METHODSThe left proximal MCA was embolized with an autologous thrombus in six beagles. Diffusion-weighted imaging (DWI) and T2-weighted imaging (T2WI) were performed every half hour during the first six hours after occlusion, followed by three time points at 12 hours, 24 hours, and one week. Perfusion-weighted imaging (PWI) and magnetic resonance angiography (MRA) were carried out at six hours, 24 hours and one week. The PWI-DWI mismatch ratio was defined as (PWI-DWI)/DWI ischemic volume.

RESULTSLacunar infarcts induced by MCA occlusion were located in the left caudate nucleus and internal capsule. All the lesions could be detected within two hours by DWI. Lesion volume on DWI increased in a time dependent manner, from (87.19 ± 67.16) mm(3) at one hour up to (368.98 ± 217.05) mm(3) at 24 hours (P = 0.009), while that on PWI gradually decreased from (7315.00 ± 2054.38) mm(3) at six hours to (4900.33 ± 1319.71) mm(3) at 24 hours and (3334.33 ± 1195.11) mm(3) at one week (P = 0.002). The mismatch ratio was 41.93 ± 22.75 at six hours after ischemia, showing "extensive mismatch", and decreased to 18.10 ± 13.74 at 24 hours (P = 0.002). No MCA recanalization was observed within 24 hours after MCA occlusion.

CONCLUSIONSLacunar infarction induced by proximal MCA occlusion could be detected early by DWI and was characterized by extensive PWI-DWI mismatch. Multimodal MRI is useful to demonstrate the natural evolution of PWI-DWI mismatch. This ischemic model could be further used for investigating early thrombolysis in lacunar stroke showing extensive mismatch.

Animals ; Diffusion Magnetic Resonance Imaging ; methods ; Dogs ; Infarction, Middle Cerebral Artery ; complications ; Magnetic Resonance Angiography ; methods ; Male ; Stroke, Lacunar ; diagnosis