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

Aim To investigate the anticoagulant efficacy and mechanism of a semi-synthesized sodiumβ-1,4-glucan sulfate (Na-MCS). Methods Anticoagulant activity was evaluated by means of coagulation assays in comparison with heparin. The anticoagulant mechanism of Na-MCS was disclosed by inhibitory analysis of the activities of coagulation factors using chromogenic substrates. Results 0. 6concentration. The dosage of Na-MCS required to double APTT of normal human plasma was 0.7represented a potent anticoagulation activity in vitro, which matched the efficacy of heparin in a certain range of concentrations. Na-MCS exhibited anticoagulant activity due to inhibition of the coagulation factors Ⅱa and Xa by the mediation of anti-thrombin AT-Ⅲ.

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 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 investigate the correlation between liver-type fatty acid binding protein (L-FABP) and nonalcoholic fatty liver disease (NAFLD) extent and other clinical parameters.Methods Ninety-six patients of NAFLD (NAFLD group) and 100 cases of healthy controls (control group) were selected.The levels of serum L-FABP and blood biochemical parameters were measured by enzyme-linked immunosorbent assay.The lesion degree was assessed by ultrasound.The body mass index (BMI),waist to hip ratio (WHR) and homeostasis model assessment insulin resistance index (HOMA-IR) were calculated.Results The WHR,BMI,fasting plasma glucose (FBG),triglycerides (TG),alanine aminotransferase (ALT),HOMA-IR and L-FABP in NAFLD group were higher than those in control group,there were statistical differences (P ＜ 0.05).The result of correlation analysis showed L-FABP level was positively related with ALT,TG,FBG,WHR,BMI,HOMA-IR (r =0.735,0.728,0.681,0.713,0.699,0.673 ;P ＜0.05),and negative correlation with HDL-C (r =-0.607,P ＜ 0.05).The L-FABP level in control group was (15.42 ± 2.51) g/L,mild NAFLD was (15.96 ± 2.92) g/L,moderate NAFLD was (17.48 ± 3.91) g/L,serious NAFLD was (25.14 ± 5.37) g/L.There was statistical difference in L-FABP level between serious NAFLD and control group,mild NAFLD,moderate NAFLD (P ＜ 0.05).Conclusion Serum L-FABP level of serious NAFLD patient significantly increases,and L-FABP level is related with biochemical parameters of liver function.

Objective:To assess clinical efficacy of micro-punch combined with electron beam radiation for the treatment of keloids on the lower jaw.Methods:Clinical data were collected from 36 patients with keloids on the lower jaw, who received micro-punch combined with electron beam radiation at Department of Dermatology, Xijing Hospital, the Fourth Military Medical University from January 2018 to January 2019, and analyzed retrospectively. There were 16 males and 20 females, with an average age of 23.80 years and a median disease duration of 32 months. The severity of keloids was evaluated before and 12 months after operation. A paired t test was used to compare keloid severity scores before and after treatment. Results:All of the 36 patients experienced primary wound healing. The keloid severity score was 7.11 ± 1.46 before operation, and significantly decreased to 2.33 ± 0.47 at 12 months after operation ( t=13.85, P=0.008) . Twelve months after the treatment, 15 patients were cured, 17 showed marked improvement, and 4 showed no response or experienced recurrence within 12 months after treatment, with a response rate of 88.89%. Conclusion:Micro-punch combined with electron beam radiation is effective for the treatment of keloids on the lower jaw.

OBJECTIVE:To establish the GC fingerpriont for Lingyang ganmao tablets.METHODS:The determination was performed on DB-5 ms capillary column.Flame ionization detector was adopted with temperature of 250 ℃ (temperature programming).The temperature of injector was 240 ℃.Carrier gas was nitrogen with flow rate of 1.0 mL/min;the sample size was 1 μL by split sampling with split ratio of 20 ∶ 1.Using peppermint ketone as reference,Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 edition) was used for the similarity analysis of 10 batches of Lingyang ganmao tablets.RESULTS:There were 14 common peaks in the batches of Lingyang ganmao tablets,similarity degrees were higher than 0.98.It was proved that the GC profiles and control fingerprint of 10 batches of samples had good consistency.CONCLUSIONS:The established fingerprint can provide reference for the identification and quality evaluation of Lingyang ganmao tablets.