BACKGROUND: An ideal marked molecule has not been found to detarmine bone marrow mesenchymal stem cells (MSCs) so as to make sure the homogenicity.OBJECTIVE: To verify the in vitro differentiation from BrdU-treced MSCs into osteoblasts.DESIGN, TIME AND SETTING: A cytological observation/n vitro was performed in Shanxi Medical University from January to October 2008.MATERIALS: Wistar rats aging 4 weeks old were provided by Experimental Animal Center of Shanxi Medical University.METHODS: MSCs were isolated and cultured by using density gradient cantrifugation combined with attachment culture method.At about 80% confluence, trypsin was used for passage and amplification. MSCs at density of 5×1010/L were inoculated in a 25-mm culture dish with L-DMDM culture medium containing dexamethasone, β -phosphoglycarol, vitamin C, and 10% fetal bovine serum. The third-passaged MSCs were labeled in vitro with 10 μmol/L BrdU tracer. Thereafter, 10 visual fields were randomly selected to calculate numbers of positive and negative ceils so as to obtain BrdU tracing rate under a fluorescence microscope (×200).MAIN OUTCOME MEASURES: Inverted microscope was used to observe cell morphology; flow cytometry was used to detect cell surface antigen, differentiation into osteoblasts, and BrdU tracing rate in vitro.RESULTS: The purified MSCs which were like fibroblasts were adherent and fusiform. The third-passaged cells were changed equidirectionally and whirlpool-arranged, and the survival rate was more than 95%. The seventh-passaged cells still grew rapidly.CD44, CD71, and CD105 expressions were positive, but CD45 expression was negative. Black particles were visualized in MSCs after Von kossa staining. BrdU tracing rate was more than 90%.CONCLUSION: Density gradient centrifugation combined with attachment culture method can effectively isolate and purify rat MSCs which are cultured in vitro for a long period and differentiated into osteoblasts. BrdU tracer is safe, effective, and convenient to successfully label MSCs.

AIM: To explore the mutual effect of co-culture of wild-type astrocytes (ASC) and motor neurons VSC4.1 (VSC) on the respective ability to produce reactive oxygen species (ROS). METHODS: The inhibition rates of cell growth in ASC and VSC in co-culture or independent culture was detected after exposed to excitatory stimulus by MTT method. Real-time observation of ROS production by ASC and VSC labeled with Hoechst 33342 was detected by confocal microscopy under the conditions of co-culture or independent culture. RESULTS: Higher concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in ASC alone, while lower concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in VSC only. Real-time observation by confocal microscopy showed that ROS production by VSC under the condition of co-culture, which showed a notable increase at 15 min, was significant less than that in independent culture, which peaked at 5 min and was gradually decreased. ROS production by ASC in co-culture began to increase significantly at 10 min. CONCLUSION: Compared to independent culture, ASC reduces the resting ROS production by co-cultured with VSC, while ASC prolongs the duration of ROS production by VSC after exposed to excitatory stimulus.

Purpose To explore the difference of expression and prognostic significance of SP1,KLF4 and p21 in low grade ovarian serous carcinoma (LGSC) and high grade ovarian serous carcinoma (HGSC).Methods The expression of SP1,KLF4 and p21 protein was examined with immunohistochemistry EliVision method in cases with LGSC and HGSC.Kaplan-Meier analysis and Cox multivariate survival analysis were used to assess the impact of SP1,KLF4 and p21 expression on prognosis of LGSC and HGSC.Results SP1,KLF4 and p21 expression were detected respectively in 74.5％,17.0％ and 11.7％ HG-SC cases,and in 65.2％,34.8％ and 26.1％ LGSC cases.Compared to control group,the expression level of SP1 was significantly higher (P ＜ 0.05),but the expression level of KLF4 and p21 were significantly lower (P ＜0.05).There was no significant difference of SP1,KLF4 and p21 expression between HGSC and LGSC (P ＞ 0.05).The expression of SP1,KLF4 and p21 were associated with FIGO stage,meanwhile SP1 associated with residual tumor size in HGSC (P ＜ 0.05).There was a significant negative correlation between SP1 and KLF4,p21 proteins in HGSC (P ＜ 0.05).Kaplan-Meier analysis revealed that there were significantly poor overall survival (OS) of 5 years for patients with HGSC displaying high expression of SP1,or low expression of KLF4 and p21 (P ＜0.05),but no significantly improved OS for patients with LGSC (P ＞ 0.05).Cox analysis showed that SP1 overexpression is an independent prognosis factor for HGSC.Conclusion Overexpression of SP1 and low expresion of KLF4 and p21 contribute to carcinogenesis of HGSC and LGSC.They are associated with a poor prognosis of HGSC,but not LGSC,meanwhile SP1 is an independant prognosis factor for HGSC.

Considering the weak relevance between the stages of C,D,I and O and the weak operability of CDIO theory in teaching,mind map is introduced into CDIO theory to drive medical English teaching.Firstly,how mind map and CDIO theory are mutually complementary is discussed,based on which,along the main clue of CDIO theory,a further discussion is carried out from four levels:conceiving the main structure and content of a passage,designing sentence teaching,implementing vocabulary memorization and operating self-directed learning,all with mind map as an assistance.Moreover,specific cases are used to exemplify how mind map is applied to CDIO medical English teaching.Using.mind maps in CDIO medical English teaching is highly coherent and operable.It helps to integrate the four stages of C,D,I and O and enhances teaching quality and learning efficiency.

Objective To investigate the correlation between hemoglobin level and health status of the elderly living in communities in Beijing.Methods A random cluster sampling method was used to select residents living in communities of Beijing city,and a cross-sectional study was carried out by questionnaires,scene testing and blood sample collection.WHO-formulated criteria were applied for diagnosing anemia.The health indicators in questionnaires included visual impairment,physical disability,decreased health,self-care,fatigue,anorexia,independent walking distance,exercise frequency,intelligence status and computing power.Results Complete information was obtained in a total of 1 948 elderly people,including 790 cases of male and 1 158 cases of female,with an average age of(73.9±6.1)years and a median age of 74 years(65-100).The mean level of hemoglobin in the 1 948 people was(135.65 ± 14.48) g/L,with (142.56 ± 15.56) g/L in male and (130.95 ± 11.53) g/L in female.Hemoglobin level was significantly lower in female than in men (t =54.739,P＜ 0.01).Hemoglobin level was decreased with aging,and negatively associated with appetite,physical strength,walk assistance,visual acuity and physical ability(r=-0.055,-0.067,-0.071,-0.114,-0.095;P =0.022,0.005,0.004,0.000,0.000),while positively associated with health status,activities in daily life,athletic ability,exercise frequency and intelligence (r =0.073,0.126,0.122,0.066,0.124;P =0.002,0.000,0.000,0.006,0.000).Conclusions The hemoglobin level of the elderly decreases with aging and is associated with health status and quality of life in the elderly,which should be taken care seriously.

Through the expert questionnaire (Delphi Indagate) to form Traditional Chinese Medicine (TCM) Healthy Lifestyle Guiding Principles.We conducted two rounds of surveys of TCM experts in the field of preventive health care on the basis of previous literature research.This study was aimed to form the framework in order to entry to the TCM Healthy Life-style Guiding Principles.The results showed that the positive coefficient of experts in two rounds of investigation were 84.31％ and 100％,respectively.The authoritative decree were both more than 0.7.The expert opinion score mean median was 9.21 and 9.085.Median mean values were 7.965 and 7.925.The median scores of full marks were higher than 65.12％ and 66.67％.The minimum coefficients of variation were 0.11 and 0.10.It was concluded that in the investigation and study,the enthusiasm of the experts,professional standards and credibility were high.Expert opinion is more concentrated.Expert opinion is well coordinated and gradually converges through two rounds of surveys.Through the Delphi indagate,the basic framework and items of TCM guiding principles of healthy lifestyle were established,which laid the foundation for further standardizing the guiding principles of TCM in healthy lifestyle.

Objective: To observe the killing and clearance effect of an Er:YAG laser combined with sodium hypochlorite on Enterococcus faecalis at different depths of the root canal in vitro to provide a reference for clinical application. Methods : A total of 75 models of Enterococcus faecalis infection were successfully established and randomly divided into three groups (25 per group). Ten random samples per group underwent no processing. The remaining models in group A were treated with an Er:YAG laser combined with sodium hypochlorite 52.5 g/L. Group B was treated with 52.5 g/L sodium hypochlorite (positive control), and group C was treated with normal saline (negative control). The bacterial reduction rate was calculated for each group. The bactericidal effect on the surface of the root canal wall was observed in 5 samples by scanning electron microscopy. Results: There was no statistically significant difference (P > 0.05) between group A (100.00 ± 0.00) and group B (98.62 ± 2.01) but was a statistically significant difference between group A (100.00 ± 0.00) and group C (64.37 ± 2.45) (P < 0.05). The percentage reduction in bacteria was higher in group A (99.46 ± 2.31) than in groups B (92.89 ± 3.07) and C (56.72 ± 4.96) (P < 0.05). The decrease in bacteria was greater in group A (97.62 ± 3.73) than in groups B (72.49 ± 2.35) and C (44.42 ± 4.78) (P < 0.05). The bacterial reduction rate in group A (95.89 ± 2.46) was higher than that in groups B (63.88 ± 1.08) and C (33.31 ± 5.21) (P < 0.05). There were significant differences at different dentin depths in the bacterial reduction rates among groups A, B and C (P < 0.05). Electron microscopy analysis showed that the Er:YAG laser combined with 52.5 g/L sodium hypochlorite was better than either treatment alone at removing bacteria from the surface of the root canal wall. Conclusion An Er:YAG laser can effectively enhance the bactericidal effect of sodium hypochlorite on the inner wall of the root canal.