Objective To investigate the protective effects of recombinant human erythropoietin(rhEPO)treatment on histopathologic changes seen in hyperoxia induced lung injury.Methods Rat pups were randomly divided into four groups:Ⅰ:air-exposed control group,Ⅱ: air-exposed+rhEPO-treated group,Ⅲ:hyperoxia-exposed control group,Ⅳ:hyperoxia-exposed+rhEPO-treated group.GroupⅢ and Ⅳ rats were exposed to 85% oxygen.GroupⅡand Ⅳ rats were received rhEPO (1 200 U/kg) subcutaneously on postnatal 0 day and 2 day.On postnatal 14 day,survival curve,measurement of body weight and lung weight,radical alveolar counts(RAC),microvessel count were compared,CD_ 31 and vascular endothelial growth factor(VEGF) were performed by immunostaining to assess hyperoxia-induced changes in lung morphology.Results Treatment of hyperoxia-exposed rats with rhEPO prolonged the survival and resulted in a significant increase in the weight gain of body and lung[(25.88?2.59) vs(18.8?3.93) P

Objective To observe temporal expression of matrix metalloproteinases(MMP-13) and tissue inhibitor of metalloproteina-ses-1 (TIMP-1) in lung of newborn rats with hyperoxia induced chronic lung disease (CLD),and to explore the relationship of CLD with MMPs.Methods The neonatal rats within 24 hours after birth were randomly divided into hyperoxia-exposed group(n=40) and control group(n=40).On postnatal 1,3,7,14 and 21 days,lung tissue of rats in 2 groups were collected.Lung histological changes were evaluated by hematoxylin and eosin and Masson stain;Collagen Ⅰ was detected by enzyme linked immunoadsorbent assay;MMP-13 and TIMP-1 were identifide by immunohistochemistry.Results Exposured to hyperoxia enviroment for 21 days,the number of alveolar decreased,terminal air space enlarged,inter-alveolar septa thickened,and deposition of interstitial collagen fibers.On 14 and 21 days,collagen Ⅰ in the lung of hyperoxia-exposed group increased significantly compared with that of control group(P0.05),obviously decreased on 21 day(P

Acupuncture Therapy ; Deglutition Disorders ; etiology ; therapy ; Humans ; Male ; Middle Aged ; Stroke ; complications

Objective To evaluate the left ventricular diastolic function in coronary artery disease (CAD)patients without regional wall motion abnormality by real-time three-dimensional specke tracking imaging(RT3D-STI),and to investigate the value of RT3D-STI in detecting CAD.Methods According on coronary angiography,92 patients without regional wall motion abnormality were divided into two groups:control group(n = 21 ),without coronary artery stenosis;case group (n = 50 ),coronary stenosis group. Two dimensional images and the three-dimensional full volume dynamic images of left ventricle were collected.Routine parameters and end-systolic left ventricular globle longitudinal strain (GLS),globle circumferential strain (GCS),globle area strain (GAS),globle radial strain (GRS)and corresponding strain imaging diastolic index (SI-DI)were aquired,respectively.The parameters by RT3D-STI were analyzed to diagnose coronary artery stenosis.Results Compared with control group,there were no significant difference in routine parameters,GLS,GRS,GAS and GCS(P >0.05 ).Longitudinal SI-DI(L-SI-DI),area SI-DI(A-SI-DI),radial SI-DI (R-SI-DI),circumferential SI-DI (C-SI-DI)in case group were significantly reduced compared with control group (P <0.05).The sensitivity of L-SI-DI,A-SI-DI,R-SI-DI and C-SI-DI were 90.5%,81 .0%,85.7%,81 .0%,and the specificity of them were 82.0%,78.0%,68%,56%, respectively,L-SI-DI was the best in sensitivity and specificity.L-SI-DI,A-SI-DI,R-SI-DI were significantly correlated with Gensini score (r = -0.706,-0.547,-0.410 P < 0.05 ).Conclusions RT3D-STI can quantitatively assess the early changes of left ventricular diastolic function,which has important significance for early diagnosis of coronary artery stenosis.

PurposeTo analyze the far infrared thermography characteristics before and after surgery in patients with acoustic neuroma.Materials and Methods Thirty-two patients with acoustic neuroma were included as observation group and 40 normal healthy individuals as control group. Un-cooled thermal imaging system (ATIR-M301B) was used with working temperature of 20-25℃. Craniofacial infrared images were collected to analyze temperature differences among different detection zones.Results Far infrared thermography revealed that there were no obvious temperature differences between both sides of supraorbital region, endocanthion region, frontal region and buccal region in the control group (P>0.05). The temperature differences in bilaterally symmetrical parts of supraorbital region, endocanthion region, frontal region, buccal region were significantly higher in observation group (t=1.557, 1.714, 1.483 and 1.569,P<0.05). The craniofacial temperatures of 32 patients changed after operation, and the differences reduced in supraorbital region, endocanthion region, frontal region, and buccal region (t=2.655, 2.462, 2.897 and 4.465,P<0.05).Conclusion Far-infrared thermography inspection can detect abnormal temperature changes.

C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.

There were significant increase in urine protein excretion,raised malondialdehyde(MDA) level and expressions of NF-?B,p22phox and p47phox in renal tissue,and significant decrease in reduced glutathione,superoxide dismutase,vitamin C and E levels in type 2 diabetic Goto-Kakisaki rats after 12 weeks. There was obvious histomorphologic change in the kidneys.All the above indices were improved by intraperitoneal injection of?-lipoic acid(35 mg/kg q.o.d).Besides,significant positive correlations were found of MDA level to p22phox,p47phox and NF-?B in the renal tissue,?-lipoic acid seems to protect the diabetic kidney in this diabetic rat model via antioxidative effects.

Objective To construct wtp53/junB fusion gene and its eukaryotic expression vector in order to provide the basis for further application of polygene union therapy in hepatocellular carcinoma. Methods Polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR) and gene recombination techniques were used to construct the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the enhanced green fluorescent protein (EGFP). The transfection of pEGFP-C1-wtp53/junB in hepatoma HepG2 cells was detected by the location of green fluorescence. Results The DNA sequence of wtp53/junB fusion gene was successfully cloned into the pEGFP-C1 plasmid and the sequence was the same as what we expected. Green fluorescence located on cell nucleus proved that pEGFP-C1-wtp53/junB was transfected into HepG2 cell line successfully. Conclusion We successfully constructed the eukaryotic vector of pEGFP-C1-wtp53/junB fusion gene, which carries the EGFP, and transfects it into human hepatoma cell nucleus. It may lay the basis for studying the synergetic effect of wtp53 and junB in hepatocellular carcinoma.

OBJECTIVEBronchopulmonary dysplasia (BPD) is a multifactorial disease resulting from the impact of injury (including oxygen toxicity, barotrauma, volutrauma, and infection) on the immature lung. Oxygen toxicity is thought to be a major contributing factor in the pathogenesis in BPD. Previous animal studies have shown that exposure to hyperoxia in the neonatal period causes lung structural changes that are similar to the histology seen in human infants with BPD. Erythropoietin (EPO) has pleiotropic actions including antioxidant, anti-apoptotic, anti-inflammatory and angiogenic effects. Animal experiments reveal that EPO may have protective effects on hyperoxic lung injury, but the mechanisms remain unknown. The aim of the study was to evaluate the anti-inflammatory effects and understand mechanism of action of EPO on the hyperoxia-induced BPD in newborn rats.

METHODSeveral litters of Wistar pups were pooled together within 12 hours after birth and randomly divided into four groups: I. air-exposed control group, II. air-exposed human recombinant erythropoietin (rhEPO)-treated group, III. hyperoxia-exposed placebo group and IV. hyperoxia-exposed rhEPO-treated group . Group III and IV rats were exposed to 85% oxygen. Group II and IV rats received rhEPO (1200 IU/kg) subcutaneously on postnatal days 0 and 2. Group I and III received 0.9% saline in the same way. Pups from each group were sacrificed on days 3, 7, and 14. Blood hemoglobin concentration, hematocrit and platelet count were determined by blood cell analyzer. Total protein content in bronchoalveolar lavage fluid (BALF) and myeloperoxidase (MPO) were measured by biochemical assay. Changes of monocyte chemoattractant protein-1 (MCP-1) and cytokine-induced neutrophil chemoattractant-1 (CINC-1) mRNA expressions were measured by RT-PCR.

RESULTIn group III, there were a few inflammatory cells infiltrations in interstitium on day 3 and inflammatory response worsened on day 7. Alveolar and capillary hypoplasia and interstitial fibrosis were evident on day 14. The pathological changes were ameliorated greatly in group IV and the survival was prolonged. There were no abnormal raises of hemoglobin concentration, hematocrit and platelet count in group IV compared with group I. Total protein concentration in BALF was measured as a marker for capillary leakage. MPO is a major constituent of neutrophil cytoplasmic granules and its activity therefore is a direct measure of neutrophil presence and an indirect indicator of lung injury. Total protein concentration and MPO in BALF were greatly depressed in group IV compared with group III, P<0.05, P<0.001. The upregulation of genes of CINC-1 and MCP-1 was closely related with lung inflammation caused by oxidative stress. MCP-1 and CINC-1 mRNA expression were detected and it was found that their changes were in line with the degree of lung inflammation. MCP-1 and CINC-1 mRNA expression increased in group III compared with group I especially on day 7, P<0.01 or <0.001. The changes of MCP-1 and CINC-1 mRNA were positively correlated with changes of MPO in BALF covering all groups on days 3, 7 and 14, respectively (r = 0.391, P<0.05; r = 0.701, P<0.01; r = 0.600, P<0.01; r = 0.471, P<0.01; r = 0.789, P<0.01; r = 0.588, P<0.01).

CONCLUSIONEPO could significantly reduce the lung inflammatory cell infiltration, and capillary endothelial cell injury in hyperoxic lung injury in newborn rats. The mechanism may be related with the inhibition of MCP-1 and CINC-1 gene expression by EPO.

Animals ; Animals, Newborn ; Chemokine CCL2 ; metabolism ; Disease Models, Animal ; Erythropoietin ; pharmacology ; Female ; Hyperoxia ; complications ; pathology ; Lung ; drug effects ; metabolism ; pathology ; Lung Injury ; etiology ; metabolism ; pathology ; Male ; Rats ; Rats, Wistar ; Recombinant Proteins

OBJECTIVETo evaluate remineralization efficacy of an arginine containing dentifrice on initial enamel carious lesions in vitro.

METHODSHuman enamel blocks with early lesions were prepared and randomly divided into three treatment groups: negative control group (distilled and deionized water), positive control group (fluoride containing dentifrice and 0.14% sodium monofluorophosphate), and test dentifrice group (8.0% arginine and 0.14% sodium monofluorophosphate). The lesions were subjected to a pH-cycling regime for 10 days. Surface enamel microhardness of the enamel blocks from each group was measured before and after pH cycling, and the surface microhardness recovery was calculated. Then, specimens were analyzed for enamel fluoride uptake (EFU) through acid etching method, after which they were treated in demineralization solution for a 2 h period of acid challenge. The other specimens were sectioned and examined through polarized light microscopy.

RESULTSIn the test dentifrice group, microhardness recovery and EFU were significantly higher than those in the negative control and positive groups. The test dentifrice group was significantly resistant to the acid challenge compared with the other groups. Conspicuous remineralization of enamel subsurface lesions was observed under polarized light microscopy among samples treated with test dentifrice, whereas the control groups showed no significant changes on enamel subsurface lesions.

CONCLUSIONThis study presents the potential superiority of Pro-Argin dentifrice over conventional fluoride dentifrice in promoting the remineralization of initial enamel lesions.

Arginine ; Calcium Carbonate ; Cariostatic Agents ; Dental Caries ; Dental Enamel ; Dentifrices ; Fluorides ; Hardness ; Humans ; Phosphates ; Tooth Demineralization ; Tooth Remineralization