Purpose To investigate the expression and clinical significance of Foxp3 ( cell surface marker of regulatroy T) mRNA and its protein in endometrial carcinomas and normal endometrial tissues. Methods Real-time fluorescence quantitative PCR and immuno-histochemical SP methods were used to detect the expressions of mRNA and protein in tumor tissue of 84 cases of endometrial carcino-mas and 40 cases of normal endometrial tissue, then to analyze the relationship between Foxp3 gene and clinical pathological character-istics of endometrial carcinoma specimens, such as differentiation, FIGO stage. Results Foxp3 mRNA and it′s protein expression of endometrial carcinomas were significantly higher than that of normal endometrial tissues. There were significantly relationships between Foxp3 mRNA expression and FIGO stage of endometrial cancer, Foxp3 mRNA expressions of III+IV stage was higher than that ofⅠ+Ⅱ stage endometrial carcinoma (P<0. 05). But the relationship between Foxp3 expression and differentiation degree reached differ-ent conclusions in the two detection methods. By immunohistochemistry the expression of Foxp3 protein was correlation with histological differentiation grade (rs =0. 72, P <0. 01). In poorly differentiated endometrial carcinoma Foxp3 + cell number was significantly higher than that in well differentiated endometrial carcinoma. By detection of real-time fluorescence quantitative PCR method, Foxp3 mRNA expression was not correlated with tumor grade (rs =0. 01, P=0. 35). Conclusion Foxp3 in endometrial carcinomas are high expressions. Immunohistochemical method has more clinical value than real-time fluorescence quantitative PCR test results. Foxp3 may be involved in the regulation of the development of endometrial cancers.

BACKGROUND:Our previous work has demonstrated that bone marrow mesenchymal stem cels (BMSCs) transplantation can improve the heart function of rats with myocardial infarction. However, the overal efficacy is not satisfactory. OBJECTIVE: To adopt pioglitazone as a peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist combined with BMSCs transplantation therapy, thereby further improving cardiac function of rats with myocardial infarction as wel as investigating the relevant mechanisms. METHODS:Twenty Sprague-Dawley rats with myocardial infarction were induced by the left anterior descending coronary artery ligation. The animals were randomized into two groups: BMSCs and BMSCs+pioglitazone. Two weeks later, al the animals received the injection of BMSCs labeled with PKH26 in PBS into the local infarct zone, and then pioglitazone (3 mg/kg/d) was given by the oral gavage for 2 weeks in the BMSCs+pioglitazone group after the cel transplantation. After 2 weeks of cel transplantation, cardiac functions were evaluated by echocardiography. The expressions of PPAR-γ, Connexin 43 and molecules in TGF-β1/SMAD signaling pathway were examined in different areas of the left ventricle from each harvested heart using immunofluorescent staining, western blot assay and qRT-PCR. RESULTS AND CONCLUSION:There were no differences in the baseline parameters of cardiac function between the two groups. At 2 weeks after cel transplantation, the left ventricular internal diameter at end-diastole, left ventricular internal diameter at end-systole and left ventricular ejection fraction were significantly improved in the BMSCs+ pioglitazone group; the expressions of PPAR-γ and Connexin 43 were distinctly increased in different zones of the left ventricle; the levels of TGF-β1, SMAD2 and SMAD3 were obviously attenuated in the infarct zone and border zone. The above-mentioned findings suggest that pioglitazone, a PPAR-γ agonist, can enhance BMSCs potential in improvingthe heart function after myocardial infarction, and PPAR-γ may elevate the expression of Connexin 43via the blockade of the TGF-β1/SMAD signaling pathway in the procedure.

Tumor angiogenesis plays a pivotal role in the progress of tumor. Among the various endogenous angiogenic inhibitors discovered, the human plasminogen kringle 5 (K5) has been demonstrated to be a potential inhibitor of the proliferation and migration of vascular endothelial cells in vitro. The replication-incompetent adenovirus (Ad) vector Adeno-X-CMV-K5 (Ad-K5) (where CMV is cytomegalovirus) was constructed and its antiangiogenic effect was tested on vascular endothelial cell and tumor cell. For the construction, the K5 cDNA was fused in-frame with human plasminogen signal sequence and inserted into the eukaryotic expression vector pcDNA3 to form pcDNA3K5. The recombinant plasmid was subcloned into the shuttle plasmid pShuttle under the control of the constitutive CMV immediate-early promoter. The plasmid carrying the cDNA for K5 (pShuttleKS) was then recombined with the Adeno-X viral DNA and transformed into E. coli DH5alpha. The resultant recombinant plasmid pAd-K5 was transfected into human embryonic kidney (HEK) 293 cells with liposome. The adenovirus expressing human plasminogen kringle 5 (Ad-K5) was successfully packaged and propagated in 293 cells, as detected by the cytopathic effect (CPE) on the cells, and the viral titer in the supernatant was 5 x 10(8) pfu/mL by plaque assay. Both human umbilical vein endothelial cell line ECV304 and human breast carcinoma cell line MDA-MB-231 were infected with Ad-K5 and Ad-LacZ, which was used the negative control, and assayed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Compared with uninfected control and Ad-LacZ infected control, Ad-K5 infected group at 80 MOI (multiplicity of infection) significantly inhibited ECV304 proliferation; the difference between uninfected control and Ad-LacZ infected control was not significant. In contrast, there was no significant difference in the proliferation of MDA-MB-231 among all the treatments. In addition, the Ad-K5 at 100 MOI inhibited the differentiation and tube formation of ECV304 on ECMatrix gel. These results suggested that the recombinant replication-defective Adenovirus expressing human plasminogen kringle 5 inhibited the proliferation, differentiation and tube formation of ECV304 and had no effect on the proliferation of MDA-MB-231. Adenovirus mediated human plasminogen kringle 5 gene therapy may be a potential treatment of cancer through angiogenesis inhibition.
Adenoviridae ; genetics ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; genetics ; Humans ; Neovascularization, Physiologic ; genetics ; physiology ; Peptide Fragments ; genetics ; physiology ; Plasminogen ; genetics ; physiology ; Polymerase Chain Reaction

Objective To evaluate the efficacy of sevoflurane in preventing depression-like behavior in mice and the relationship with brain-derived neurotrophic factor (BDNF)/tyrosine kinase B (TrkB) signaling pathway.Methods Forty-four clean-grade male C57BL/6 mice,weighing 18-22 g,aged 8-10 weeks,were divided into 2 groups (n =22 each) using a random number table method:control group (group C) and sevoflurane group (group S).Mice in group C inhaled oxygen for 30 min,and mice in group S inhaled 2.5％ sevoflurane for 30 min.The forced swimming test and novelty-suppressed feeding test were performed after the mice were fully awake.The brains were immediately removed under anesthesia at the end of inhalation of oxygen or sevoflurane,and the prefrontal cortex and hippocampus were isolated for detection of the expression of BDNF,TrkB and phosphorylated TrkB (p-TrkB) by Western blot.Results Compared to group C,the immobility time and feeding latency were significantly shortened,the expression of p-TrkB in the prefrontal cortex and hippocampus was up-regulated (P＜0.05),and no significant change was found in the feeding consumption or expression of BDNF and TrkB in the prefrontal cortex and hippocampus in group S (P＞0.05).Conclusion Sevoflurane produces a preventive effect on depression-like behavior in mice,and the mechanism is related to increased phosphorylation of TrkB in BDNF/TrkB signaling pathway.

OBJECTIVETo evaluate the efficacy of the interferon alpha-2b nasal spray in prevention of rubella and measles virus infections.

METHODSThe properly selected volunteer groups have been divided into interferon alpha-2b experimental and control group. The experimental group received interferon alpha-2b treatment by nasal spray for 2 days before the immunization, then both groups were challenged with rubella and measles attenuated live vaccine respectively through nasal spray. The sera from pre-immunization and 21 and 28 days after immunization were collected to test the IgG antibody titers. The influence on the viral antibody titer reflects the viral preventive effect by interferon alpha-2b.

RESULTSThe antibody titer difference of measles virus between experimental and control group was 1.26 (21 day) and 2.96 (28 day), there were statistically difference between them; the difference of rubella virus was 0.95 (21 day) and 0.37 (28 day), but there were no statistically differences found.

CONCLUSIONThe preliminary results showed that the interferon alpha-2b can be used as prevention method for measles and rubella viral infections.

Administration, Intranasal ; Adult ; Antibodies, Viral ; blood ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Male ; Measles ; immunology ; prevention & control ; virology ; Measles Vaccine ; immunology ; therapeutic use ; Measles virus ; drug effects ; immunology ; Recombinant Proteins ; Rubella ; immunology ; prevention & control ; virology ; Rubella Vaccine ; immunology ; therapeutic use ; Rubella virus ; drug effects ; immunology ; Treatment Outcome ; Vaccination ; methods ; Vaccines, Attenuated ; immunology ; therapeutic use ; Young Adult

BACKGROUNDTo study the molecular mechanism of interferon alpha2b(IFNalpha2b) inhibiting the SARS virus replication. SARS-associated coronavirus (SARS virus) cDNA chip was developed and applied to detect the virus RNA transcription levels in the interferon-treated and untreated cell cultures, and the mechanism of anti-SARS virus activity of interferon alpha2b in cell culture system was explored.

METHODSSARS virus cDNA chip was prepared by comparing the published SARS virus genome sequence, and the cDNA chip was used to study the interferon alpha2b function during SARS virus replication.

RESULTSSARS virus cDNA chip was successfully prepared by using PCR method. The results showed that the cDNA chip could be used to detect the viral RNA transcription level. Interferon alpha2b could inhibit almost all the SARS virus gene transcription. An unknown gene at the position 28130-28426 bp, named as U gene, may play an important role during the viral replication.

CONCLUSIONA SARS virus whole genome cDNA chip was established. It could be used to study the virus molecular biology and antiviral drug screening. The results also showed that interferon alpha2b could inhibit almost the whole virus gene transcription by using the cDNA chip.

Humans ; Interferon-alpha ; pharmacology ; Oligonucleotide Array Sequence Analysis ; methods ; RNA, Viral ; genetics ; Recombinant Proteins ; SARS Virus ; drug effects ; genetics ; isolation & purification ; physiology ; Severe Acute Respiratory Syndrome ; virology ; Virus Replication ; drug effects

BACKGROUNDTo study the anti-SARS virus activities of different recombinant human interferons on the cell culture system.

METHODSAnti-SARS virus activities of interferons were determined by using CPE inhibition test in human skeletal muscle sarcoma (Rda) cell culture.

RESULTSThe average minimum amount of interferon alpha 2b, alpha 1b, beta 1b or omega 1b to inhibit 50% CPE in Rda cell culture was (160.5+/-129.5) IU/ml, (149.0+/-71.7) IU/ml, (69.5+/-61.5) IU/ml, (87.3+/-47.1) IU/ml, respectively or (0.6+/-0.5) ng/ml, (10.6+/-5.1) ng/ml, (3.5+/-3.1) ng/ml, (0.9+/-0.5) ng/ml, respectively.

CONCLUSIONAll the tested recombinant interferons showed anti-SARS virus activities on the Rda cell culture with different sensitivities.

Antiviral Agents ; pharmacology ; Cell Line, Tumor ; Humans ; Interferon Type I ; pharmacology ; Interferon-alpha ; pharmacology ; Recombinant Proteins ; SARS Virus ; drug effects ; Severe Acute Respiratory Syndrome ; drug therapy ; virology

OBJECTIVETo assess the value of (99m)Tc-N-NOET ((99m)Tc-N-ethoxy-N-ethyl dithiocarbamato-nitrito) myocardial perfusion SPECT for the diagnosis of coronary artery disease.

METHODSA total of 42 patients [mean age (54 +/- 9) years, 35 men] with suspected chest pain were included in this study. 740 MBq of (99m)Tc-N-NOET was injected intravenously during bicycle exercise when the heart rate attained reached more than 85% of the expected maximum, or in cases of angina pectoris, severe arrhythmias and ischemic ST segment changes. (99m)Tc-N-NOET 740 MBq, SPECT myocardial imaging acquisitions were obtained at 15 minutes and 2 hours after (99m)Tc-N-NOET injection. Coronary angiography was performed in all patients.

RESULTSCoronary artery stenosis was detected in 26 patients and normal coronary angiography was shown in 16 patients. (99m)Tc-N-NOET myocardial perfusion imaging was abnormal in twenty-one patients out of the 26 patients with significant coronary artery stenosis (sensitivity, 81%); 14 out of 16 patients with normal angiography had a normal myocardial perfusion imaging (specificity, 88%). The positive predictive value, negative predictive value and predictive accuracy of (99m)Tc-N-NOET myocardial perfusion imaging for detection of CAD was 91%, 74% and 83%, respectively. The sensitivity of the imaging for detecting single vessel, double vessels and triple vessels disease were 60% (6/10), 86% (6/7) and 100% (9/9), respectively. There was mild (99m)Tc-N-NOET lung uptake in patients with coronary artery stenosis 15 minutes post (99m)Tc-N-NOET injection.

CONCLUSIONSPECT myocardial perfusion imaging with (99m)Tc-N-NOET supplied an important diagnostic tool for detecting coronary artery disease. Lung uptake with stress (99m)Tc-N-NOET might be related to coronary artery disease.

Adult ; Aged ; Coronary Artery Disease ; diagnostic imaging ; Exercise Test ; Female ; Humans ; Male ; Middle Aged ; Organotechnetium Compounds ; Thiocarbamates ; Tomography, Emission-Computed, Single-Photon

BACKGROUND: Chronic progressive external ophthalmoplegia (CPEO) is a mitochondrial encephalomyopathy caused by multiple mtDNA abnormalities. There is little information about the changes of ocular fundus with CPEO. The aim of this work was to measure and evaluate changes in the macular retinal thickness and optic nerve head in patients with CPEO using spectral-domain optical coherence tomography and to compare the findings with those of healthy individuals. METHODS: Totally, 18 CPEO patients were enrolled in this study. Healthy volunteers matched for gender, age, and diopter settings were included as a control group. The retinal thickness of macular central fovea, inner and outer retinal layer thickness of perifoveal macular, optic nerve head parameters, and peripapillay retinal nerve fiber layer thickness (pRNFLT) for all included cases were measured using spectral-domain optical coherence tomography. A paired t test was used to compare the differences in the studied parameters between the two groups. The correlations between macular retinal thickness, pRNFLT, disease duration, and age of onset were also analyzed. RESULTS: Among the macular parameters, retinal thickness of macular central fovea (t = -2.135, P < 0.05) and outer retinal layer thickness (t = -1.994, P < 0.05) of patients in the CPEO group were statistically significant lower than those of patients in the normal control group. For the optic nerve head parameters, the patients in the CPEO group showed a larger rim volume (t = -2.499, P < 0.05) and nerve head volume (t = -2.103, P < 0.05). The overall pRNFLT of patients in the CPEO group was statistically significant lower than that of patients in the control group (t = -4.125, P < 0.05). The comparison of pRNFLT in eight sectors showed that the pRNFLT of patients in the CPEO group was statistically significant lower than that of the control group mainly in the inferior and temporal sectors. The degree of pRNFL defect negatively correlated with the disease duration (r = -0.583, P < 0.05). CONCLUSIONS The retinal thickness of patients with CPEO was significantly thinner, which was mostly the outer retina. The patients' optic discs had a low volume and the loss of the retinal nerve fiber layer was obvious. With the extension of the disease duration, the retinal nerve fiber layer defect was even more significant.

Background:: Chronic progressive external ophthalmoplegia (CPEO) is a mitochondrial encephalomyopathy caused by multiple mtDNA abnormalities. There is little information about the changes of ocular fundus with CPEO. The aim of this work was to measure and evaluate changes in the macular retinal thickness and optic nerve head in patients with CPEO using spectral-domain optical coherence tomography and to compare the findings with those of healthy individuals. Methods:: Totally, 18 CPEO patients were enrolled in this study. Healthy volunteers matched for gender, age, and diopter settings were included as a control group. The retinal thickness of macular central fovea, inner and outer retinal layer thickness of perifoveal macular, optic nerve head parameters, and peripapillay retinal nerve fiber layer thickness (pRNFLT) for all included cases were measured using spectral-domain optical coherence tomography. A paired t test was used to compare the differences in the studied parameters between the two groups. The correlations between macular retinal thickness, pRNFLT, disease duration, and age of onset were also analyzed. Results:: Among the macular parameters, retinal thickness of macular central fovea (t = -2.135, P < 0.05) and outer retinal layer thickness (t = -1.994, P < 0.05) of patients in the CPEO group were statistically significant lower than those of patients in the normal control group. For the optic nerve head parameters, the patients in the CPEO group showed a larger rim volume (t = -2.499, P < 0.05) and nerve head volume (t = -2.103, P < 0.05). The overall pRNFLT of patients in the CPEO group was statistically significant lower than that of patients in the control group (t = -4.125, P < 0.05). The comparison of pRNFLT in eight sectors showed that the pRNFLT of patients in the CPEO group was statistically significant lower than that of the control group mainly in the inferior and temporal sectors. The degree of pRNFL defect negatively correlated with the disease duration (r = -0.583, P < 0.05). Conclusions: The retinal thickness of patients with CPEO was significantly thinner, which was mostly the outer retina. The patients’ optic discs had a low volume and the loss of the retinal nerve fiber layer was obvious. With the extension of the disease duration, the retinal nerve fiber layer defect was even more significant.