Objective To study the effect of different concentration of tumor necrosis factor-? (TNF-?) on expression of the receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin (OPG) mRNA on human dental follicle cells in vitro, and investigate the role of TNF-? in osteoclast formation during tooth eruption. Methods The 5th passage of primary cultured human dental follicle cells were treated with 0 (control group), 5, 10, 25, 50 and 100ng/ml TNF-?, respectively, for 6 hours. Total RNA was then isolated from human dental follicle cells and subjected to RANKL and OPG mRNA assay by reverse transcription-polymerase chain reaction (RT-PCR) technique. The relative expression levels of RANKL and OPG mRNA were normalized to ?-actin gene expression. Results The mRNA expression of OPG in human dental follicle cells with 5ng/ml TNF-? treatment was down regulated significantly compared with that in control group (P

Objective:To study the effects of colony-stimulating factor-1 (CSF-1) on the expression of osteoprotegerin (OPG ) in human dental follicle cells.Methods:Human dental follicle cells culture was established and cell crawling slides were stained by OPG antibody. The third to sixth passage cells were incubated with 25 ng/ml CSF-1 for 0.5,1,3 and 6 h respectively,then the conditioned medium and cells were collected for ELISA and RT-PCR assay respectively.Results:OPG was expressed in the cytoplasm of human dental follicle cells.GSF-1 at 25 ng/ml and 1 h-exposure down-regulated OPG expression in the cells(P

Objective: To investigate the expression of CSF-1 and TGF-?1 in periodontal tissue of third premolar germ during second primary molar orthodontic movement in dog. Methods: Orthodontic appliance with the force of 0.784 N for the mesial movement of mandible second primary molar was used in 4 dogs for 14 d.Then the samples including mandibular second and third molars and surrounding tissues were prepared for CSF-1 and TGF-?1 detection by immunohistochemical staining technique. Results: Strong CSF-1, TGF-?1 expression was found in the dental follicle and alveolar bone of mesial part of third premolar germ, and weak in the dental follicle and alveolar bone of distal part of third premolar germ in the treated side of the dogs. Conclusion: CSF-1 and TGF-?1 may play roles in the bone remodeling for orthodontic primary tooth movement.

To observe whether there is an interaction between hepatitis virus B (HBV) and tumor suppressor p53, plasmid PCMVp53 was transfected or cotransfected with pCMVHBV a (wild type HBV) or PCMVHBV b (mutation type HBV) into the hepatoma cell line 7721 by phosphate calcium precipitation. Apoptosis cells were labeled by annexin Ⅴ FITC and detected by flow cytometry. Another experiment was performed by cotransfecting the cells with reporter plasmid p21 luc in each group mentioned above, then the luciferase activity was measured. The results showed that the cells transfected by pCMVp53 alone exhibited high luciferase activity and high apoptosis rate. Meanwhile, the luciferase activity and apoptosis rate were further higher in cells cotransfected by pCMVp53 and PCMVHBV a , but remained unchanged in cells cotransfected by PCMVp53 and PCMVHBV b. The results indicated that P53 could induce of 7721 cell apoptosis by activating p21 transcription, and such effect could be enhanced by HBV.

BACKGROUND:Previous studies have applied long-bone mechanical separation method to obtain osteoclasts, which are terminal y differentiated cells and cannot further proliferate. Therefore a large number of osteoclasts can be harvested with bone marrow cells inducing culture method and RAW264.7 cells inducing culture method to meet clinical requirements.
OBJECTIVE:To investigate the optimal method of receptor activator of nuclear factor kappa-B ligand (RANKL) induced osteoclast precursors to differentiate into mature osteoclasts.
METHODS:After bone marrow cells were isolated from mouse, RANKL and macrophage colony stimulating factor were added into the medium together, or RAW264.7 cells were cultured with RANKL to induce osteoclasts. The osteoclast precursors were treated with different concentrations of RANKL to observe the number of generated osteoclasts and evaluate the dose-effect relationship between RANKL and osteoclastogenesis. Annexin V-FITC and propidium iodide staining were used for flow cytometry to analyze the mononuclear-macrophage apoptosis during osteoclastogenesis.
RESULTS AND CONCLUSION:When 10μg/L RANKL was used, the peak of osteoclastogenesis appeared at days 6-7;when the concentration of RANKL was up to 100μg/L, the peak appeared at days 4-5. The number of new osteoclasts was dose-dependent on the RANKL concentration. 50μg/L of RANKL was the turning point in the fitted curve from osteoclastogenesis and RANKL concentration. After the RANKL concentration was more than 150μg/L, the number of osteoclasts slowed down obviously. RANKL can induce monocyte-macrophage to form osteoclasts and promote monocyte-macrophage apoptosis. The highest number of osteoclasts would be obtained to each unit of RANKL when monocyte-macrophage cells were cultured with 30μg/L of RANKL in the same vaccination density (104/cm2). Experimental findings indicate that, RAW264.7 cells or bone marrow cells inducing culture methods are simple and feasible, the optimum cellseeding density was 104/cm2;the appropriate stimulation concentration of RANKL was 30-50μg/L.

Bronchi ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Middle Aged ; Pneumonectomy ; Pulmonary Sclerosing Hemangioma ; pathology ; surgery

Objective To know the rural doctors' knowledge,belief and practice on iodine deficiency disorders(IDD),as well as their capability on health education communication in Ye County,Henan Province.Methods Semi-structured interviews were conducted among 50 rural doctors in Ye County,Henan Province.Topics covered include:sales of salt purchases,daily supervision and monitoring activities of salt,the health effects of salt,the situation of rural doctors trained,the situation of rural doctors participated in health education and related recommendations.Results ①Twenty-nine of the rural doctors knew exactly the advantage of salt iodization.②Thirty-nine of the rural doctors had taken part in the half quantitative detection of salt once or twice a year and 15 of them knew the supervision activities of salt industry department.③Forty-nine of the rural doctors believed salt iodization was beneficial to health and 48 of them were willing to participate in health education on IDD.④The major ways of their acquiring health knowledge were promotional materials [54.0％ (27/50)],TV [50.0％ (25/50)],and lecture/training [24.0％ (12/50)].⑤Thirty-seven of the rural doctors had taken part in health education on IDD.They were used to use booklets [68.0％ (34/50)],promotional slogan [36.0％ (18/50)],broadcast [24.0％ (12/50)],and doctor consultation [24.0％ (12/50)].⑥Twenty-eight of the rural doctors thought the current health education on IDD was better.They hoped to reduce the price of salt,strengthen supervision,receive more training and health education on IDD.Conclusions The rural doctors remain relatively lack of knowledge on IDD.Rural doctors have the initiative and responsibility to conduct health education on IDD,but their pattern is still a one-way propagation.

Objective To investigate the consumption rate of non-iodized salt,and evaluate the iodine status and goiter prevalence among school children in high water iodine areas of Henan Province from 2014 to 2015.Methods In the 20 counties with high water iodine,one township was randomly selected from each location (east,west,south,north and middle) in each county;secondly,4 villages were selected from each chosen township;thirdly,15 households were selected to collect salt samples from each chosen village.In the 10 chosen counties,one village with high water iodine was selected and water samples were collected;one school was sequentially selected from the chosen village and 100 school children aged 8-10 were chosen to collect their urine samples and measure their thyroid volume.Salt iodine was tested by semi-quantitative method;iodine contents of urine and water were tested by arsenic cerium catalytic spectrophometry;thyroid volume was measured by ultrasound method.Results In the 20 counties with high water iodine,4 440 salt samples were collected and tested both in 2014 and 2015;the rates of non-iodized salt were 98.2％ (4 363/4 440) in 2014 and 98.3％ (4 366/4 440) in 2015.In the 10 chosen counties,the median water iodine contents from the chosen villages were 202.0 μg/L in 2014 and 235.0 μg/L in 2015;970and 999 urine samples of the students from the chosen villages were collected and tested in 2014 and 2015,and the median urinary iodine contents were 251.9 μg/L in 2014 and 290.6 μg/L in 2015;937 and 948 students were examined in 2014 and 2015,the goiter rates were 3.4％ (32/937) in 2014 and 7.8％ (74/948) in 2015.Stratified by water iodine,the urinary iodine contents and goiter rates of school children increased with the rise of water iodine content.When the water iodine content exceeded 300 μg/L,goiter rate of school children was 8.4％,which was higher than other groups (P ＜ 0.05).Conclusions After stopping the supple of iodized salt in high water iodine areas,the current iodine status and goiter rate of school children are still higher than normal levels.Both noniodized salt supply and water improvement to reduce water iodine content should be taken in the areas with water iodine higher than 150 μg/L.

Objective To study corrective method for displacement in the procedure of electronic portal imaging device (EPID)-based intensity-modulated radiotherapy dose valuation by studying the relative mechanical displacement of different vendor EPID (aS1000,Varian; aS500,Varian; iViewGT,Elekta).Methods A 5 cm × 5 cm field was set up to acquire portal images for three kinds of EPID,then a in house software was used to analysis the portal images.The relative displacement was acquired via analyzing a series of comparation between center positions of gantry angle ranging from 0° to 360° and gantry angle at 0°.Results In the lateral direction,the maximum relative displacement of EPID with aS1000,S500 and iViewGT were (-0.23 ±0.17) mm,(2.94±0.17) mm and (0.35 ±0.09) mm,respectively.In the longitude direction,the displacements were (-4.16 ± 0.20) mm,(-4.15 ± 0.25) mm and (-1.66 ±0.11) mm,respectively.As to longitude direction,the displacements could be well fitted with the usage of quadruplicate empiric function.Conclusions There is a significant difference at the aspect of relative displacement between different vendors EPID at different gantry angles.And the displacement in the longitude direction is obviously larger than in the lateral direction.The relative displacement should be corrected when applying EPID to the intensity-modulated radiotherapy dose verification at different gantry angles.

Objective To evaluate the effect of collagen shield delivering fluconazole eye drops made by centrifugal method on treating fungal keratitis of rabbit eyes.Methods Using centrifugal method to make corneal collagen shield,after soaked with fluconazole eye drops,the collagen shields were used to treat fungal keratitis of rabbit eyes.Results The healing focus time of collagen shield delivering fluconazole eye drops group was average 11 days,the healing focus time of only delivering fluconazole eye drops group was average 18 days.The cure rate and effective rate of 2 groups were 78.94%,63.15% and 92.10%,78.94%,there were significance between 2 groups(P<0.05).Conclusion It was innovation for corneal collagen shield delivering fluconazole eye drops made by centrifugal method to cure fungal keratitis,which had broad development foreground.