The effects of all-trans retinoic acid (ATRA) on primary gastric carcinoma models made by subcutaneous implanting gastric cancer to mice were observed. Thirty-two cancer bearing nude mice were randomly divided into 4 groups: control group (group Ⅰ)， ATRA low dose feeding group (100μg/day， group Ⅱ), moderate dose feeding group (300μg/day， group Ⅲ), and high dose feeding group (1 000 μg/day, group Ⅳ). The alteration of tumor growth, morphology, cytobiology, and immuno-histochemical assay were perfermed. The results showed significant inhibition of tumor growth and inducing differentiation in the group Ⅲ and group Ⅳ as compared with group Ⅰ (Ｐ＜0.01),and inhibited expression of p53, p21 protein in implanted tumor. The authors consider that ATRA has some effects of growth inhibition and differentiation on gastric cancer cells in vivo, and these is related to the inhibition of expression p53 and p21 onco-gene of cancer cells and accelerate apoptosis of carcinoma cells.
Objective To investigate the role of urotensin Ⅱ(UⅡ) in the pathogenesis of asthma.Methods Lung function，differential cell count and level of UⅡin induced sputum were studied in 26 asthmatic patients in acute exacerbation and in clinical remission.Results Induced sputum UⅡ level from acute asthma was higher than that of remissed asthma [(58.88±47.38)pg/mL vs (12.69±12.78)pg/mL，Plt;0.01].Induced sputum UⅡ levels of asthma patients in acute exacerbation had a tendency to increase as disease deteriorated，which negatively correlated with FEV1% predicted (r=-0.326，Plt;0.05)，but did not with sputum total cell and neutrophil counts(Pgt;0.05).No significant difference of induced sputum UⅡ levels was found between male and female，smokers and non-smokers.Conclusion UⅡ may play a role in acute exacerbation of asthma
Objective To investigate the clinical characteristics of neutrophilic asthma ( NA) .Methods NA patients were collected from the out-patient and in-patient departments of Respiratory Diseases of Xinqiao Hospital between January 2010 and December 2010. The results of the medical records,pulmonary function tests, and induced sputum cytology were analyzed retrospectively. Results The NA patients with neutrophil percent ≥ 61% accounted for 33. 1% ( 51 /154 ) of all the asthmatics patients detected by induced sputumin the same period, and 45 cases with complete records were included. Of the 45 cases recruited, 20 cases ( 44. 4% ) were in-patients,2 cases ( 4. 4% ) were with controlled asthma, 3 cases ( 6. 7% ) were with cough variant asthma, 30 cases ( 66. 7% ) were female, 12 cases ( 26. 7% ) were atopic patients, and 27 cases ( 60% ) had acute exacerbation. The age of onset of 35 patients ( 77. 8% ) were after 12 years. FEV1% pred lt; 60% and gt; 80% was obtained in 55. 9% ( 19/34) and 14. 7% ( 5 /34) of patients respectively. The result of bronchodilator test was positive in 64% ( 16/25) of patients, and mean increase in FEV1 was 11. 7% . The percentage of neutrophil and eosinophil was ( 74. 5 ±9. 1) % and ( 2. 4 ±2. 5) % respectively in induced sputum, and 35. 6% ( 16/45) of the patients had increased eosinophil percentage ( gt;3% ) . Conclusions In our study, most of NA is severe and acute exacerbation asthma, and its clinical features are various. The mechanismand clinical significance of increased neutrophils in asthmatic patients are unclear and more studies are needed.
Objective To review new progress of related research of bone tissue engineering in recent years. Methods Domestic and international l iterature concerning bone tissue engineering was reviewed and analyzed. Results In the recent years, great progression had been made in the research and development of bone tissue engineering, it had been used in more and more hospitals, and relevant national regulations and protocols had been set up. As to seed cells of bone tissue engineering, autologous and allogeneic stem cells had been widely used, while recently embryonic stem cells and induced pluri potent stem cells had attracted most attentions. In the field of scaffolds materials, significant improvementshad been made, from natural extractions to artificial polymers; from single construction to multiple compounds with surface modifications. As to the methods of construction, the static seeding approach had been widely accepted, and the appl ications of bioreactor had provided a stable and various micro-enviroment for the vitro-culture of different stem cells, which had beenregarded as an alternative way of vitro-culture and construction for bone tissue engineering. Conclusion With the tremendous help of the techniques and approaches above, we shall expect a promising future of a new generation bone tissue engineering based medical products in the years to come.
Objective To study the effect of rat osteoblast conditioned culture medium on the BMSCs differentiation of allogeneic rat and to find a new approach to provide seed cells for bone tissue engineering. Methods BMSCs and osteoblasts were harvested from 10 healthy one-week-old SD rats (male and female, weighing 20-30 g) by adherent method and enzyme digestion method respectively. Cell identification was conducted. Osteoblast conditioned culture medium was prepared by mixing supernatant of osteoblasts at passage 1-5 with complete medium (1:1). Then, BMSCs at passage 2 were co-cultured with osteoblast conditioned culture medium (inducement group) and complete medium (control group), respectively. The morphological changes of co-cultured BMSCs were observed by inverted phase contrast microscope, the growth condition of BMSCs was detected by MTT method, the expressions of ALP, Col I and osteocalcin (OCN) in the cocultured BMSCs were tested by immunohistochemistry staining, and the expressions of Col I and OCN mRNA were detected by RT-PCR. Results In the inducement group, BMSCs grew bigger, changing from long fusiform to flat and polygon with protuberance 7 days after co-culture; the presence of cell colony-l ike growth was observed 9 days after co-culture. Cell growth curve demonstrated that the counts of BMSCs was increased with time, there were more cells in the control group than that of the inducement group, and there was a significant difference in cell counts between the control and the inducement group 4-7 days after co-culture (P lt; 0.05). For the inducement group, ALP staining was positive 12 days after co-culture, the calcium nodules were appeared 18 days after co-culture, Col I and OCN were positive 21 days after co-culture, and the expressions of Col I and OCN mRNA were detected by RT-PCR 21 days after co-culture. Conclusion Rat osteoblast conditioned culture medium can significantly induce the differentiation of allogeneic rats’ BMSCs towards osteoblasts.
Objective To investigate the effect of NGF on fracture heal ing, and to study the role of BMP-2 induced osteoblast. Methods Sixty cleaned male Kunming mice (aging 6-8 weeks and weighing 23-25 g) were made fracture models in the middle of femoral shaft and randomly divided into four groups (groups A, B, C and D, n=15). Fracture was treated with NGF/ normal sal ine, BMP-2, BMP-2 /NGF/normal sal ine, and normal sal ine in groups A, B, C and D, respectively. After 14, 21 and 28 days, the specimens were selected from 5 mice each group to do the biochemical and histological analysis. Beforethe mice were killed, the arteriovenous blood was taken from their eye-ball to test the ALP activity. Results After 14 days,21 days and 28 days, the gross observation showed that the size and hardness of bone tissue, and callus tissue growth increased in groups A, B and C order and were higher than those in group D; the X-ray films showed that the calcified area increased in groups A, B and C order and were higher than those in group D; the histological observation showed that the trabecular maturity increased in groups A, B and C order and were higher than those in group D. The osteoblast area, the gray degree value of the radiographs in callus tissue, the ALP contents of serum and callus tissue, calcium content of callus tissue and net weight of callus were higher in groups A, B and C than in group D. There were significant differences (P lt; 0.05) in osteoblast area and gray degree values of the radiographs at 14, 21 and 28 days; in ALP contents of serum at 14 days; in ALP contents of callus tissue at 14 days and 21 days; in calcium content of callus tissue at 21 days and 28 days among 4 groups. There were significant differences in net weight of callus between groups B, C and groups A, D at 14 days (P lt; 0.05). At 21 days and 28 days, the trabecular surface index of osteoblast, the average trabecular volume and the mean trabecular width decreased as time went on, having an increase order of groups A, B, C and was higher in groups A, B, C than in group D, showing significant differences among 4 groups (P lt; 0.05). Conclusion NGF promotes the heal ing of fractures. NGF possesses synergistic effect on ectopic bone formation induced by BMP-2.
Objective To explore effect of platelet-rich plasma (PRP) on rabbit BMSCs differentiation into SC in vitro and to detect secretory function of the differentiated cells. Methods BMSCs isolated from 5 mL bone marrow of 2-montholdNew Zealand white rabbit were cultured using density gradient centrifugation and adherence screening methods. A total of 5 mL femoral vein blood was obtained from rabbits to prepare PRP using modified Appel method. The BMSCs at passage 3 were divided into three groups: the combined induction group, in which the cells were cultured with complete medium containing PRP after β-mercaptoethanol and retinoic acid inductions; the simple induction group, in which the cells were cultured with L-DMEM complete medium without PRP afterβ-mercaptoethanol and retinoic acid induction; the control group, in which the cells were cultured with L-DMEM complete medium. Growth condition of the cells in each group was observed using inverted microscope. cell identification was conducted at 4, 7, 9, and 11 days after culture using immunofluorescence staining method, and NGF content was detected by ELISA method. NGF mRNA expression was assayed by RT-PCR 11 days after culture. Results Most cells in the combined induction and the simple induction group were out of BMSCs typical cell morphology 4 days after culture; cells in the combined induction group were out of BMSCs typical cell morphology and changed into cells resembl ing SC in terms of morphology and contour 9 days after culture. The cells in the control group showed no obvious morphological changes. S-100 protein expression in the cells was evident in the combined induction and the simple induction group at each time point after induced culture; the positive expression rate of cell in each group was increased over time, and significant differences were evident between the combined induction group and the simple induction group 7, 9, and 11 days after culture (P lt; 0.05). Control groupwas negative for the expression. There were significant differences when comparing the control group with the combined induction group or the simple induction group in terms of NGF content at each time point (P lt; 0.01). Significant difference was evident between the combined induction group and the simple induction group 7, 9, and 11 days after culture (P lt; 0.05), and no significant difference was noted 4 days after culture (P gt; 0.05). Relative intensity of NGF mRNA expression in the combined induction group was greater than that of the simple induction group 11 days after culture (P lt; 0.05). Conclusion Rabbit BMSCs can differentiate into SC excreting NGF under certain induction condition in vitro. PRP can remarkably promote BMSCs differentiation into SC.
Objective To investigate the possibility of ectomesenchymal stem cell of human embryo facial process in differentiating into osteoblasts.Methods Ectomesenchymal stem cells of human embryo facial process were isolated and cultured in mineralized promoting solution containing 10 mmol/L β-glycerophosphate, 100 μg/ml ascorbic acid and 10 nmol/L dexamethasone supplemented with 15% FBS. The morphological change was observed by phase contrast microscopy. The characteristics of cells was identified by immunohistochemistry assay. Alkaline phosphatase activity was tested and the form of mineralized nodules was tested with Von Kossa staining. The expression of osteocalcin was identified by RT-PCR.Results There were significant changes in the shape of the cells after 3 days cultured in mineralized promoting solution. The cells became larger and the shape changed from fibroblast-like to multilateral. The result for anticollogen typeⅠstaining was positive. The alkaline phosphatase activity increased. Mineralized nodules were formed aftercultured 25 days by Von Kossa staining. RT-PCR assay showed induced cells expressed osteocalcin.Conclusion Ectomesenchymal stem cells of humanembryo facial process can be induced to differentiate into osteoblasts by mineralized promoting solution.
OBJECTIVE To testify the inductive osteogenesis of allogeneic bone matrix gelatin (BMG) in promoting intervertebral fusion. METHODS The gelatin sponge, allogeneic BMG, decalcified bone matrix (DBM) and alcohol conserved bone were implanted respectively into the intervertebral space of rabbit, whose intervertebral discs were removed before implantation. The intervertebral spaces were evaluated by X-ray and histological examination at 4, 8, and 12 weeks after operation. RESULTS No obvious immune rejection was observed. Amounts of new bone were formed in the intervertebral spaces at 4 and 8 weeks. And complete infusion of the intervertebral spaces were appeared at 12 weeks. CONCLUSION Allogeneic BMG can promote bone fusion of intervertebral spaces through osteoinduction, which suggests that allogeneic BMP is an ideal substitute for bone replacement.
Osteoinducin is a kind of new biomaterial, which contains bone morphogenic protein (BMP). Its role of inducing osteosis has been confirmed by laboratory study. Since 1992, osteoinducin was applied to treat tumorous defects of bone for 7 cases. After eradication of the lesion of tumor, stickor granulelike osteoinducin was filled in the defects. In 8 weeks, the bone defects had the sign of osteosis on roentgenogram. No rejection reaction and local inflammation were found. The property, operation procedure and indication were introduced. Compared with other artificial bones, the osteoinducin showed faster osteosis, better histoco mpatibility and more convenient usage. It was a good material for bone graft.