BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are in advantages of easy collection and amplification in vitro, but the culture and induction in vitro still need to be modified. OBJECTIVE: To investigate the differentiated potency of BMSCs into ostcoblasts by modified in vitro culture methods and inductor matching. DESIGN: Observational study. SETTING: Department of Medical Laboratory, General Hospital of Guangzhou Military Area Command of Chinese PLA. MATERIALS: This study was performed in the Stem Cell Tissue Engineering Laboratory, Department of Medical Laboratory, Genera Hospital of Guangzhou Military Area Command of Chinese PLA from July 2004 to June 2006. Twenty adult SD rats of clean grade, irrespective of gender, weighing 140-180 g were provided by Animal Experimental Center, General Hospital of Guangzhou Military Area Command of Chinese PLA. The experimental animals were disposed according to ethical criteria. Β-sodium glycerophosphate, dexamethasone, and vitamin C were provided by Sigma Company, USA; goat-anti-rat osteocalcin antibody by DSL Company, USA; S-P immunohistochemical kit by Maixin Biotechnology Developing Co., Ltd., Fujian. METHODS: Cells were cultured and induced into osteoblasts by modified methods. Separation and culture of BMSCs: By anesthesia, bone marrow of bilateral femur and tibia was separated to prepare single cell suspension; subsequently, the suspension was inoculated in culture bottle, and the culture media was semi-quantitatively changed after 48 and 96 hours in order to get rid of non-adherent hematopoietic cells. The liquid was changed every three days to further get rid of non-adherent cells. At 80% cell confluence, the medium was digested with 0.25% trypsin and cells were subcultured in the ratio of 1:2. MSCs in the second generation were inoculated in 6-well culture plate and glass fiat plate; after 48 hours, basic culture fluid was removed. Differentiation of BMSCs into osteoblasts: Subcultured BMSCs differentiated into osteoblasts induced by dexamethasone (10 mmol/L), β-sodium glycerophosphate (10-7 mol/L), and vitamin C (50 mg/L). MAIN OUTCOME MEASURES: Ten days after differentiation by modified culture methods and inductor matching, alkaline phosphatase activity was measured with Ca-Co technique. Twelve days after culture, osteocalcin secretion was detected with immunohistochemical method. Two weeks after culture, cell mineralization was detected with Von kossa staining. RESULTS: Alkaline phosphatase activity: Alkaline phosphatase staining of cells was apparent; gray-black particles or massive precipitations were observed in cytoplasm after positive reaction; regions expressing alkaline phosphatase activity were brown-black. Osteocalcin secretion: Osteocalcin was apparently positive; nucleus was blue; cytoplasm was brown. Cell mineralization: Induced cells grew layer by layer, and adiaphanous nodus was observed at the same time. Black mineralized nodus granules in various sizes were observed after Von kossa staining, and this suggested that mineralized apposition occurred. CONCLUSION: BMSCs may be successfully cultured and induced into osteoblasts by modified culture method.

BACKGROUND:With the potential of multi-directional differentiation and high proliferation, bone marrow mesenchymal stem cells (BMSCs) have wide application prospect in tissue engineering. OBJECTIVE:To investigate changes in cells and bioactivity in the differentiation from BMSCs into osteoblasts. DESIGN, TIME AND SETTING:A randomized control experiment was performed at the Laboratory of Stem Cells and Tissue Engineering, Department of Medical Experiment, General Hospital of Guangzhou Military Area Command of Chinese PLA from July 2004 to June 2006. MATERIALS:Twenty adult SD rats of both genders were used for bone marrow collection. METHODS:Rats were equally and randomly divided into an experimental group and a control group. BMSCs were isolated from adult rats by modified bone marrow culture method. BMSCs were inoculated in basic medium containing 10% fetal bovine serum, high glucose DMEM, 100 U/L penicillin, 100 U/L streptomycin and 2 mmol/L glutamine in the control group. BMSCs were inoculated in conditioned medium (basic medium, 10 mmol/L β-glycerophosphoric sodium, 10-7 mol/L dexamethasone, 50 mg/L vitamin C). Cell slide was prepared. MAIN OUTCOME MEASURES:Inverted microscope and transmission electron microscope were used to observe cell appearance. Gomori modified calcium-cobalt method was applied to do alkaline phosphatase staining. Immunocytochemistry was employed to measure osteocalcin expression. RESULTS:Forty-eight hours after inoculation, a mass of BMSCs adhered to a wall. Seventy-two hours later, BMSCs proliferated and well grew. Seven to nine days later, cells were grown to 80% confluency. Transmission electron microscope showed BMSCs with big cell nucleus and immature appearance. After in vitro osteoblast induction, BMSCs proliferated, aggregated, had node and mineralized. One week later, alkaline phosphatase activities were expressed in BMSCs. Two weeks later, osteocalcin expression was detected. CONCLUSION:After one week of in vitro osteogenic induction, BMSCs enter the process of osteoblast transformation, remain proliferative activities, and can be used as seed cells in bone tissue engineering

Objective To investigate the clinical and serological manifestations of the interstitial lung disease(ILD) resulting from the dermatomyositis(DM) or the polymyodsitis(PM),and the factors affecting the resulting of treatment for the purpose of providing necessary reference for clinical treatment and diagnosis.Methods The clinical manifestations and the findings of serological tests of 18 patients compared with only DM or PM.Results ILD was likely to strike 42.9 percent of patients with DM or PM,who were higher than those without ILD in the rate of positive Jo-1,LDH and Reynolds disease.Conclusion The patients with DM or PM,with their high probability to be stricken by ILD,need HR-CT scanning in order to be diagnosed as early as possible.

Objective To explore the relationship between TGF-?, P21ras, c-fos and the apoptosis of human embryonic pancreatic islet cells. Methods Semi-quantitative RT-PCR and immunohistochemistry (SABC method) were used to detect TGF-?, P21ras, c-fos expression at different growth periods of the primary human embryonic pancreatic islet cells and human fibroblast cells. Results P21ras expression was weak in every period of the pancreatic islet cells and significantly weaker than the fibroblast cells at exponential growth period and was stronger than in incubation period and retention period (P

Objective To explore the influence of age factor on the treatment of cutaneous hemangioma with (90Sr-90Y) applicator.Methods A total of 224 babies and children of different ages with cutaneous hemangioma were treated by using 90Sr-90 Y applicator.Then,the relation between their therapeutic effects and ages were analyzed respectively.Results The differences in the therapeutic effect between group of one month to less than one year and group of one to less than five years,and group of one month to less than one year and group of five to ten years,and between group of one to less than five ages and group of five to ten years (P＜0.05),were statistically significant.With an increase of the age,the recovery rate gradually decreased,but their effective rate,improvement rate and ineffective rate increased.In the 199 well-healed or effective patients,the differences in the distribution of treatment course between group of one month to less than one year and group of one to less than five years,and group of one month to less than one year and group of five to ten years,were statistically significant (P＜0.05); but the difference in the distribution of treatment course between group of one to less than five ages and group of five to ten years was not statistically significant (P＞0.05).The number of treatment course presented an increased tendency with an increase of the age for patients.Conclusions The age of patients is a factor influencing the therapeutic effect on cutaneous hemangioma with 90 Sr-90Y applicator.The younger the patients,the better their therapeutic effects,the shorter their courses of treatment,and the smaller the adverse reaction of beta ray.

Objective To assess the image quality (IQ) of an iterative reconstruction (IR) technique (iDose4) from prospective electrocardiography (ECG)-triggered coronary CTA on a 256 MSCT scanner and determine the optimal dose reduction using IR that can provide IQ comparable to filtered back projection (FBP).Methods Prospectively ECG gated CCTA were performed on 120 patients [76 men,44 women; age:(53 ± 10)y] using a 256-slice MSCT (Brilliance iCT,Philips Healthcare).The control group (Group A,n =30) were scanned using the conventional tube output (120 kVp,210 mAs) and reconstructed using FBP.The other 3 groups were scanned with the same kVp but successively reduced tube output as follows:B (n =30):105 mAs,C (n =30):84 mAs:D (n =30):65 mAs and reconstructed using IR levels of L4 to L6,respectively.All images were reconstructed using the same kernel (XCB).Two radiologists graded IQ in a blinded fashion on a 4-point scale (4-excellent,3-good,2-fair and 1-poor).Quantitative measurements of CT values,image noise,Signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were obtained in each group.Analysis of variance (ANOVA) was used for comparisons of objective evaluation indices (noise,CNR) and radiation dose (CTDIvol,DLP,ED) between the four groups.The Kruskal-Wallis test was used for comparisons of demographic data and for detection of differences in subjective evaluation of IQ among groups.A level of P ＜ 0.05 was considered statistically significant.A ROC analysis was performed to determine a radiation reduction threshold up to which excellent IQ was maintained.Results There was no significant differences in objective noise among Groups A (37.4 ±7.9) HU,B(33.2±7.1) HU,C(35.7±9.8) HU,and D(36.0±6.8) HU (F=1.48,P=0.22).There was no significant differences in CNR among Groups A(15.0 ±2.3),B(16.5 ±3.6),C(16.3 ±3.5),and D(15.3±2.8) (F=1.70,P =0.17).Group B and C had good and excellent scores of the subjective IQ (≥3),and there was no significant differences in the scores of the subjective IQ between Group A,and Groups B,C (P =0.30-1.00).Significant differences in image sharpness and study acceptability were observed between groups A and D (P ＜ 0.01).Using the criterion of excellent IQ (score 4),the ROC curve of dose levels and IQ acceptability established a reduction of 60％ of tube output (Group C) as optimum cutoff point (AUC:0.76,95％ CI:0.65-0.87).The effective dose (ED) of Group C was 61％ lower than that of Group A,(1.2 ± 0.1) mSv vs.(3.1 ± 0.6) mSv.Conclusion Iterative reconstruction techniques can provide 61％ ED reduction in prospectively-triggered coronary CTA using 256-slice MSCT while maintaining excellent image quality.

Objective To investigate cortical activation patterns for covert and overt picture naming with functional magnetic resonance imaging (fMRI). Methods fMRI data were collected on 24-27 years old volunteers during performance of covert and overt picture naming. After statistical postprocessing analysis, head movement data were compared across tasks and average neural activation maps were available for both tasks. Results Mean and maximal translations of head movement in covert picture naming were less than those in overt picture naming, but the difference has no statistical significance (P=0.23). It was shown that covert picture naming involved an orchestration of bilateral occipital gyri and cerebellums, bilateral supplementary motor area, postcentral gyri, inferior frontal gyrus and anterior cingulate cortex. Activations in overt picture naming included those in covert naming (but more intensive), bilateral precentral gyri and posterior superior temporal gyri, left anterior superior temporal gyrus, bilateral thalamus, basal ganglia, and left insula. Conclusion Covert and overt picture naming are two different tasks involving different neural processing networks and levels. They can not be taken as substitutes for each other.

Objective:To explore the abortion effect and mechanism induced by Chinese herbal medicine rhubarb extract.Methods:The pregnant mice at day 3 of gestation were Drenched by Aqueous extract of rhubarb at different doses (7,5,2.5 g/kg)once per day for five days,and mices in control group were oral administration of same volume of saline.All mices at 12 day of gestation were sacrificed.estradiol and progesterone levels in serum were detected by Radioimmunoassay;Content of IFN-γ, IL-2 and TNF-αin uterine lysates were measured by ELISA, macrophages in endometrium were determined by immunohistochemical method, mast cells in endometrium were tested by Toluidine blue staining, respectively.Results: showed as follows:With increasing dose of rhubarb extract,abortion rate and resorption rate were gradually increased and was 100%abortion in 7g/kg treatment groups.Estrogen and progesterone content showed a downward trend,IFN-γ,IL-2 and TNF-αin uterine lysates and the numbers of macrophages and mast cells were significantly higher in mices Drenched by Aqueous extract of rhubarb than in the control.Conclusion: These results show that rhubarb extract not only interfere stability of pregnancy state in pregnant mices because of its diarrhea,but also can directly affect endometrial environment in early embryonic mice,Resulting in abortion.

Objective To investigate hippocampal neuronal damage and dynamic change of α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor subunit GluR2 in status epilepticus, to find out whether GluR2/glyceral dehyde-3-phosphate dehydrogenase (GAPDH) interaction has any change.Methods Male Wistar rats (62 cases) were induced to status epilepticus by using LiC1-pilocarpine.The 62 rats were divided into 1 h (6 cases),6 h (12 cases), 24 h (12 cases),72 h (12 cases)and 7 d (20 cases)after status epilepticus.At the same time, the healthy control group (12 cases) was established.Morphologic changes of hippocampus and the amount of apoptotic cells in healthy control and SE model groups at different time points (6 h, 24 h, 72 h, 7 d) (6 cases each group) after status epilepticus were quantified by adopting Nissl staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling staining respectively.Expressions of GluR2 in healthy control and SE model groups at 1 h,6 h,24 h,72 h and 7 d (6 cases each group) after status epilepticus were detected by using Western blot.Co-precipitation and Western blot techniques were used to investigate whether the GluR2/GAPDH interaction in the hippocampus was increased.Results Compared with the healthy control group, the number of nerve cells in the hippocampal CA1 and CA3 regions was significantly reduced at all studied time points(F =30.866,24.043, all P ＜0.05).Apoptotic cells in hippocampal CA1 and CA3 regions were significantly increased at 24 h,72 h and 7 d after status epilepticus (F =84.762,52.574, all P ＜ 0.01).GluR2 at 1 h,6 h after status epilepticus was equal to that of the control group (P ＞ 0.05), but it was shown to be significantly down-regulated at other studied time points (F =76.506,P ＜ 0.01);when compared with the healthy control group,the GluR2/GAPDH interaction was significantly up-regulated in the hippocampus at 72 h after status epilepticus (t =7.029, P ＜ 0.05).Conclusions Status epilepticus can lead to neuronal damage in the hippocampus.Down-regulation of GluR2 and increase of the GluR2/GAPDH complex formation might be one of the mechanisms involved in hippocampal neuronal damage.

Objective To evaluate and testify the diagnostic efficacy of the ADC value of the diffusion-weighted imaging (DWI) in prostatic peripheral zone cancer. Methods There were 40 patients with prostatic diseases proved by ultrasound guided systemic biopsy. DWI scans were performed with 1.5T MR scanner using TOSORPA coil and b value of 800 s/mm2. The peripheral zones of prostate were divided in six areas and were attributed to noncancerous and cancerous areas according to the results of biopsy. The minimal ADC values of each peripheral zone were recorded and analyzed. Results There were 240 areas, in which 89 areas were cancerous and the others were noncancerous. The mean ADC value of cancerous areas was (0.98±0.30)×10-3 mm2/s, while the noncancerous areas was (1.59±0.29)×10-3 mm2/s. The diagnostic sensitivity, specificity and accuracy were 80.58%, 92.42% and 86.73% respectively with cutoff point ≤1.24×10-3 mm2/s, whereas 66.56%, 96.95% and 86.26% respectively with cutoff point ≤1.04×10-3 mm2/s. The diagnostic sensitivity was 98.63%, the specificity was 37.07%, the accuracy was 58.82% with cutoff point <1.69×10-3 mm2/s. Conclusion The ADC value ≤1.24×10-3 mm2/s should be regarded as the diagnostic standard of prostate cancer. When ADC value is between (1.04-1.24)×10-3 mm2/s, some cancer might be missed.