Objective To find out a proper way to detect green fluorescent protein (GFP). Methods Kidneys, livers and femurs from GFP transgenic mice and C57BL/6J wild type mice were employed for in vivo study.The samples were dehydrated with alcohol and acetone individually before embedding, then frozen, paraffin and resin sections were made for the detection of GFP. C3 P12 cells which derived from calvaria bone cells of GFP transgenic mouse were used for the detection of GFP in vitro. Cells were exposed to alcohol, acetone and PBS after paraformaldehyde fixation. Laser scanning microscopy was employed for GFP detection. Results In frozen sections, both kidney and liver samples which exposed to 4% buffered paraformaldehyde fixation had strong GFP signals, while GFP signal disappeared completely in fresh frozen sections without fixation. Much stronger GFP intensity was found in acetone treated samples than in alcohol treated paraffin sections, but without apparent difference in GFP intensity in acetone and alcohol treated resin samples. Acetone and alcohol made no difference in fixed C3 cells in different time courses. Conclusion Acetone treated paraffin sections are preferable for GFP detection.

Objective To study the expression changes and significance of human telomerase reverse transcriptase (hTERT)-mRNA and mutant p53 protein in thyroid carcinoma tissues in the elderly.Methods The expression level of hTERT-mRNA was examined by in situ hybridization in 50 samples of thyroid carcinoma tissues from the elderly and 41 samples of thyroid carcinoma tissues from the non-elderly and 30 samples of thyroid benign lesions.The expression level of p53 was examined by immunohistochemistry in all subjects.Results The positive rate of hTERT-mRNA was 96.0％ (48/50),85.3％ (35/41) and 10.0％ (3/30) in thyroid carcinoma tissues from the elderly,the non-elderly and thyroid benign lesions respectively,and there were significant differences between the three groups (x2=73.61,P=0.000).The positive rate of p53 was 92.0％ (46/50),85.3％ (35/41) and 13.3％ (3/30) in thyroid carcinoma tissues from the elderly,the non-elderly and thyroid benign lesions respectively,and there were significant differences between the three groups (x2 =62.30,P =0.000).Conclusions The positive rates of hTERT-mRNA and p53 in thyroid carcinoma tissues are higher in the elderly than in the non-elderly,which can be used to evaluate the biological behavior and prognosis of thyroid carcinoma in the elderly,and they play the important roles for targeted therapy.

Objectvies:To investigate the role of vascular endothelial growth factor(VEGF) in the pathogenesis of endometriosis(EMs). Methods:A total of 70 specimens of endometriosis and 30 specimens of the normal controls were evaluated by immunohistologically techique with polyclonal antibody against VEGF. Results:The expression levels of VEGF in endometriotic tissues was higher than that in the normal endometrium( P

ObjectiveTo observe the effect of sport walking on heart and lungs functions in old women.MethodsForty-two old women aged from 60 to 69 had sport walking for 4 months; the speed and intensity were controlled according to their heart rate respectively. The indexes of respiration, circulation and rheoencephalography were measured before and after exercise.ResultsAfter exercise, blood-pumping function of heart improved obviously, stroke volume increased from (65.22±11.41)ml to (72.10± 10.78)ml, ejection fraction increased from (60.10±5.03)% to (68.78±6.25)%, while heart rate declined from (77.45±8.69) times/min to (7.89±8.21)times/min, capacity increased from (2.86±0.36)L to (3.34±0.53)L, and maximal voluntary ventilation for every minute increased from (96.14± 15.21)L to (114.02±16.01)L, significantly different compared with that before treatment ( P<0.01). The fluid time of rheoencephalography reduced from (0.171±0.058)s to (0.128±0.049)s ( P<0.01).ConclusionSport walking under proper intensity can improve the function of respiration and circulation system in old women, so it is a good way to keep health for the elders.

BACKGROUND:Human amniotic epithelial cells are an important source of cells in regenerative medicine as its multipotentation, but new studies mainly focused on differentiation features and there were little research oneffect of culture in vitro on biological property of amniotic epithelial cells.
OBJECTIVE:To analyze the effects of in vitro culture on growth, cellphenotype and differentiation capacity of human amniotic epithelial cells into cardiomyocyte-like cells, and explore the correlation of primarily cultured human amniotic epithelial cells marker SSEA-4 expression level and the change of biological characteristics of human amniotic epithelial cells.
METHODS:Primarily cultured human amniotic epithelial cells were obtained from amniotic tissues by using the same separation protocol. Human amniotic epithelial cells were cultured in vitro. The proliferation, cellphenotype and the differentiation capacity of human amniotic epithelial cells into cardiomyocyte-like cells were evaluated by means of cellcounting kit-8, flow cytometry and real-time PCR.
RESULTS AND CONCLUSION:The SSEA-4 positive cells in primarily cultured human amniotic epithelial cells from different fetal tissues were between 26.7%-97%, which indicated that there was great individual difference among amniotic tissue samples. Moreover, with passage, the SSEA-4 expression in human amniotic epithelial cells decreased significantly, which did not correlate with the SSEA-4 expression in primarily cultured human amniotic epithelial cells. Results indicated that there was great individual difference in SSEA-4 expression level in primarily cultured human amniotic epithelial cells from different amniotic tissue samples. Thus, it is necessary to set up clinical screening indexes to get samples with higher SSEA-4 expression stably and to control the quality of human amniotic epithelial cells. In addition, during culture period, SSEA-4 expression level was affected by culture conditions. The culture conditions of human amniotic epithelial cells should be optimized to maintain SSEA-4 expression at a high level. In addition, the differentiation capacity of human amniotic epithelial cells into cardiomyocyte-like cells was also affected by individual difference among different samples and culture conditions, which wil be further studied in the future.

Objective To investigated to implement condition of preventive measure and control effect for endemic fluorosis in Fengshun County from 2005 to 2006.Methods It was investigated according to the National Surveillance Program of Endemic Fluorosis.Hupo,Daizai and Anquan Villages of Tangxi Town in Fengshun County were selected as monitoring spots.The usage of reforming water facilities,fluoride content in drinking water and urine of children aged 8-12 years and the prevalence rate of dental fluorosis of children were investigated.Resul tsin 2005 and 2006.a total of 18 reforming water facilities were surveyed and six of which were damaged or out of service.In 2005,the fluoride content in drinking water in the 3 villages was 2.10,1.22 and 0.15 mg/L The prevalence rate of dental fluorosis of children aged 8-12 years was 54.23%(64/118),38.91%(79/203) and 9.10%(6/66).The urine fluoride content of children was 0.95,0.90 and 1.05 mg/L,respectively.In 2006,the fluoride content in drinking water in Hupo,Daizai and Anquan Village was 2.01,1.57 and 0.21 mg/L.The prevalence rate of dental fluorosis of children aged 8-12 years was 26.47%(27/102),12.50%(23/184)and 6.15%(4/65),respectively.The urine fluoride content of children was 0.97,0.61 and 0.59 mg/L.Conclusions The outcome of surveillance data in Fengshun County has reached the sanle level as that of non-disease area.However,the management of reforming water facilities should be improved.

Objectives To investigate the prevalence of animal plague in Horqin Right Front County and provide scientific bases for prevention and treatment of the disease.Methods Plague surveillance was carried out according to the requirements of National Plague Monitoring Programme at 3 monitoring points in Horqin Right Front County in 2012.Squirrel density,the number of small rats,rat fleas,pathogenic and serological surveillance of Yersinia pestis were included in the study.Results A total of 356 dauricus were captured,the average density was 0.88 rats/hm2; female and male ratio was 1 ∶ 1.31 and the female embryo rate was 96.46％.The captured 13 small nocturnal mice belonged to 2 species,which included 10 cricetulus barabensis and 3 cricetulus longicaudatus,and the capture rate of small nocturnal mouse was 4.33％ (13/300).A total of 98 rat fleas were collected from 25 rats; rat flea carrying rate was 100％(25/25) and flea index was 3.92.Eighty rat burrows were detected,and flea index was 0.188.Pathogen test results were negative.Indirect hemagglutination test was used to examine 356 rat sera and 1 serum was positive.Conclusions Detection of positive rat sera has proved the existence of plague epidemic in Horqin Right Front County.Therefore,the work on plague prevention and control has a long way to go.

Objective To explore the clinical characteristics ,diagnosis, treatment and prognosis of phaeohyphomycosis. Methods Clinical data were collected, including history, physical examination, cranial and spinal imaging. Brain biopsy was performed. Data of the pathology and incubation of brain tissue were analyzed. Responsiveness to treatment was followed up. Results A previously healthy three and half years old boy was presented to our unit, with a three- month history of recurrent headache, vomiting, progressive paraplegia accompanied by urinary continence and constipation. A computed tomogram scan and magnetic resonance imaging of the brain revealed multiple lesions located in the region of the parietal - occipital lobes, periventricular area and frontal lobe, with prominent surrounding edema and irregular peripheral enhancement of the mass after the administration of contrast materials. A cerebral biopsy was performed and the pathological report was cerebral phaeohyphomycosis. The culture of the tissue and cerebrospinal fluid grew a same fungus identified as exo-phiala dermatitidis. The patient's response to therapy was poor, the parents of the boy gave up therapy, and the boy died 1 month later. Conclusions Cerebral Phaeohyphomycosis caused by Exophiala dermatitidis is rare, but the most serious form of fungus infection. Pathology and incubation of the tissue are essential for diagnosis. There is no curative therapy and the prognosis is poor.

Objective To investigate expression levels of epithelial mucin 1 (MUC1) and epitbelial mucin15(MUC15) in elderly patients with papillary thyroid carcinoma and assess the role of MUC1 and MUC15 in the pathogenesis of thyroid papillary carcinoma.Methods Protein expression of MUC1 and MUC15 was detected by immunohistochemistry in 10 samples from normal thyroid tissue adjacent to thyroid adenoma,57 samples from papillary thyroid carcinoma (PTC),and 14 samples from PTC in neck lymph node metastasis.Results Expression rates of MUC1 in normal thyroid tissues,thyroid papillary carcinoma,and lymph node metastatic carcinoma were 40.0％,75.4％,64.3,respectively,and the rates for MUC15 were 0,73.7％,71.4％,respectively.The positive expression rate of MUC1 was higher in PTC tissues than in normal thyroid tissues (x2 =5.10,P=0.02) and,compared with normal thyroid tissues,the positive expression rate of MUC15 increased in PTC tissues and lymph node metastatic carcinoma (x2 =12.25 and 19.75,both P＜0.05)MUC15 protein expression was higher in micro-PTC (less than or equal to 1 cm in diameter) than in carcinoma larger than 1 cm in diameter (90.9％ vs.62.9,x2 =5.48,P=0.02).MUC15 expression was higher in PTC without lymph node metastasis than in PTC with lymph node metastasis (83.8％vs.55.0％,x2 =5.55,P=0.02).MUC1 expression was positively correlated with MUC15 expression in thyroid papillary carcinoma (r=0.35,P=0.01).Conclusions MUC1 and MUC15 may have synergistic effects in the initiation and progression of PTC.MUC15 may play a role in regulating tumorigenesis of thyroid papillary carcinoma in early stages and can potentially serve as a supplementary marker in the screening of micro-thyroid papillary carcinoma.

Objective To evaluate the application of early 18F-FDG PET-CT imaging after radiofrequency ablation (RFA) of hepatic malignancies. Methods Fifteen patients with liver tumors (five hepatocellular carcinoma, ten colorectal cancer liver metastasis ) underwent RFA as part of clinical management. The lesions were all hypermetabolic on PET-CT performed within 2 weeks prior to RFA. All subjects underwent 18F-FDG PET-CT (early PET-CT) within 24 hours after RFA. Total photopenia, focal uptake, and rim-shaped uptake were regarded as complete ablation, residual tumor, and inflammation, respectively. Follow-up PET-CT scans were performed as the reference standard. Results Twelve patients showed total photopenia at the ablation site on the early PET-CT scan, and in all of these patients, total photopenia at the ablation sites was seen on the follow-up PET-CT scans. Two patients had focal uptake at the ablation sites on the early PET-CT scan, and both of these foci increased in size and intensity, which were compatible with residual tumors at the time of ablation. Only one patient had rim-shaped uptake on the early PET-CT scan. The rim-shaped uptake disappeared on PET-CT performed 3 months later, which indicated the nature of inflammation. Conclusions There is infrequent inflammatory uptake at the RFA site of liver tumors on 18F-FDG PET-CT if scanning is performed within 24 hours after ablation. Thus, early PET-CT has the potential to evaluate the efficacy of an RFA procedure by indicating tumor-free as total photopenia and residual tumors as focal uptake.