Objective To evaluate the factors which might affect the successful reopening of occluded subclavian artery. Methods During the period of Jan. 1998-March 2007, endovascular stent placement was performed in 20 patients with occluded subclavian artery, including 12 males and 8 females, aged 17-74 years (mean 53 years). The procedures were carried out by using super-slippery guide wire via anterograde or bilateral access. Results Successful reopening of occluded subclavian artery was obtained in 14 cases and the treatment ended in failure in 6 cases. In 14 patients with successful results, the mean length of diseased artery was 3.00 cm and the mean course of disease was 9.83 months, which were 2.13 cm and 27.6 months respectively in 6 cases of failure. The successful reopening rate was 80% in arthrosclerosis cases, while it was 50% in aorto-arteritis obliterans. The successful reopening rate was 100% in those cases whose occluded artery showing a sharp stump, while the successful reopening rate was 33% in those cases whose occluded artery showing a round stump, with a statistically significant difference between the two (P< 0.05). The successful rate in cases performed via anterograde access was 65%, and it was only 43% in cases performed via bilateral access. Conclusion The etiology, the duration of disease and the shape of stump bear a close relationship to the successful reopening rate, while no obvious correlation exists between the length of diseased artery and the reopening rate. Higher reopening rate can be achieved when the procedure is performed via anterograde access.

Objective To study whether noggin alone can induce human bone marrow mesenchymal stem cells (MSCs) to differentiate into neural cells for the treatment of neural degenerative diseases.Methods Human bone marrow MSCs were primarily cultured and then divided into control,Ad group of empty adenovirus vector and Ad-noggin group of adenovirus vector with noggin gene.Their morphological changes were observed with fluorescence microscopy and laser confocal microscopy,and the results of 48 h,4,7,10 d were analyzed with statistical methods.Results Both the vector made human bone marrow MSCs carry green fluorescence.Ad group didn’t show any morphological changes.While for Ad-noggin group,a few neural-like cells appeared in 48 h after transfection.The number of such cells was increased,and there were some neurites around the cells after 4 d.The cells amplified greatly after 10 d.Compared to those of 7 d after transfection,the soma and dendritic diameters of Ad-noggin group were significantly increased after 10 d (P

Background Researches showed that stem cells can rescue damaged cells through mitochondrial transfer.This mode has been used to regenerative cell-based therapy.Retinal pigment degeneration is an eye disease of retinal pigment epithelial (RPE) cell apoptosis as pathogenetic mechanism.Whether stem cells can repair the target cells by above mechanism has not been clarified.Objective This study was to investigate the influence of mitochondrial transfer on the function of RPE cells.Methods The RPE cells of Long-Evans rats were isolated and cultured and the third generation of cells were used in sequential experiment.The cells were identified by detecting the expressions of RPE65 and Bestrophin proteins with immunofluorescence stain.Mouse neural stem cells (NSCs) (C17.2 strain) with green fluorescence protein (GFP) and without GFP were cultured.Mitotracker-green and Mitotracker-red staining were separately used to labeled the mitochondria of RPE cells and NSCs.RPE cells were cocultured with NSCs,and wheat germ agglutinin (WGA) was used to mark the tunneling nanotubes (TNT) between the two kinds of cells,and then the mitochondrial migration in TNT was exhibited by the laser scanning confocal microscope.The proportion of RPE cells in different cycles was assayed after marked with propidium iodide (PI) by flow cytometry.The contents of ATP,ADP and AMP in RPE cells were detected by high performance liquid chromatography (HPLC).Results The third-generation of RPE cells grew well with the RPE65-and Bestrophinpositive rate ＞85％.The Mitotracker-red-labeled rates of NSCs and RPE cells were no less than 95％.TNT structure was seen to appear the blue fluorescence between RPE cells and NSCs 24 hours after co-cultured and the red dye mitochondria from NSCs migrated toward red dye mitochondria from RPEs with the lapse of time.The RPE cell proportion reduced in G1 phase and increased by 5％ and 2％ in the S phase and G2/M phase respectively after mitochondrial transfer than before (P=0.016,0.114,0.189).The contents of ATP,ADP and AMP in the RPE cells were (8.77 ±3.68),(2.76±0.92) and (1.07 ±0.65) μg/mg after cell co-culture,and those before co-culture were (11.29±2.29),(3.12±0.95) and (1.59± 1.22) μg/mg,without significant differences between them (P =0.370,0.668,0.553).Conclusions NSCs can transfer normal mitochondria to co-cultured RPEs via TNT structure.Mitochondrial exchange might be one of therapeutic mechanisms of NSCs recuing damaged RPE cells.

With the rapid development of medical sciences in recent years,tissue engineering technology has made great achievements,while the choosing of seed cells has been a major focus of the field.Adipose-derived cells have the advantages of rapid expansion,good stability,no immune rejection and potential of multiplex differentiation.They can differentiate into cells originated from different germ layers such as:adipocytes,osteoblasts,chondrocytes,endothelial cells,myocytes,neuronal cells and so on.Besides,adipose tissue can be harvested easily and substantially with small trauma to human body.So ADSCs are expected to be an ideal choice of seed cells in tissue engineering.This article introduces the research progress of ADSCs in osteochondral tissue engineering.

Objective To analyse the survival time and related factors of patients with brain stem glioma who received 3DCRT.Methods Thirty-six patients with brain stem tumor were admitted from October 2004 to December 2008 and all received 3D-CRT with the dosage (50-54 Gy,25-30 f,5-6 weeks).During treatment,the patients’ outcomes were analyzed by observing the changes of symptoms,signs and adverse radiotherapy reaction and all of them were followed-up in the next 3 years.The survival data were analyzed by Kaplan-Meire method.Results The median survival time was 9 months in the 23 pediatric patients and 15 months in 13 adult patients.One-,two-and three-year survival rates between pediatric group and the adult group were 43.5 ％ (10/13) vs 76.9 ％ (10/13),26.1％ (6/23) vs 46.2 ％ (6/13),8.7 ％ (2/23) vs 38.5 ％ (5/13).Karnofsky performance scale score at admission (x2 =20.059,P =0.000),tumor site (x2 =17.585,P =0.000),growth pattern (x2 =21.247,P =0.000) were associate with survival time.Conclusion 3DCRT is an effective therapy to brain stem glioma,childhood onset,pontine glioma,diffusion style and Karnofsky performance scale less than 80 are risk factors of poor prognosis.

Objective To research the influences of gambogic acid on rats'cytochrome P450 isoforms by using the Cocktail probe drugs. Methods Rats were randomly divided into several groups. One group of rats received gambogic acid solution containing 0.5 %CMC-Na (2 mg?mL-1,once per day) orally for six days. At the same time,the rats in the other group received 0.5 %CMC-Na solution serving as blank control group. Blood samples were collect at different time point,and the plasma concentration of cocktail probe drugs was determined by HPLC. Results There was insignificant difference of caffeine and dapsone metabolism between the gambogic acid group and the blank group,but the chlorzoxazone was eliminated faster in the gambogic acid group,and the half life of chlozoxazone was shorter. Conclusion Gambogic acid has no significant influence on the cytochrome P450 isoforms of CYP1A2 and CYP3A4,but has an induction on the CYP2E1.

Objective To examine the expression of Noggin in CNS of the developing rat. Methods In situ hybridization histochemistry(ISHH) was performed using digoxigenin-labeled cRNA as probes. Results It was revealed that densely and deeply stained noggin positive cells were detected in cortex,hippocampus,cerebellum,and nucleus of hypothalamus and thalamus in embryonic day(E)16 rats.The number of noggin positive cells was increased in the thalamus and medulla oblongata at postnatal day(P)1-2,whereas decreased in the hippocampus and cortex.The number of noggin positive cells was decreased significantly in brain at 1 week postnatal(P1W),and began to increase at P2W,especially in the cortex and hippocamps.Strong positive signal can be detected in the frontal cortex,parietal cortex,cingulated cortex,hippocampus,olfactory and cerebellum at 1 month postnatal(P1M).The expression of noggin begins to decline at P3M,only sparse noggin positive cells can be seen in CNS at P18M.Furthermore,there is no noggin positive cells seen in the spinal cord of rats during development.Conclusion Our results indicated that noggin could play an important role in CNS development of rats.

Objective To examine the expression of BMP4 in CNS of the developing rat. Methods In situ hybridization histochemistry(ISHH) was carried out on tissue sections using specific digoxigenin\|labeled oligonucleotide probe. Results It showed that BMP4 mRNA positive cells were located mainly in cerebellum and olfactory at E16.Strong positive signal was seen in hypoglossal nucleus,and moderate signal also seen in spinocerebellar tract and spinal lemniscus at P1\|2.The number of BMP4 mRNA positive cells was increased in the frontal cortex,parietal cortex,and hippocampus subiculum at P1W.The peak of BMP4 expression was in cortex and periamydaloid cortex.Widely distributed BMP4 mRNA positive cells were detected in cortex and hippocampus of rats at P1M,strong positive signal was observed in temporal CNS at P3M,strong positive signal was observed in hippocampus,temporal corex and periamydaloid cortex,lateral nucleus of thalamus and paraventricular nucleus of hypothalamus.BMP4 mRNA positive cells were also found in corex,hippocampus,hypothalamus and thalamus at P18M.Conclusion\ These results indicated that BMP4 could play an important role in CNS development of rats.

BACKGROUND: The study about the multiple differentiation potentials of the mesenchymal stem cells is still on the stage of the animal experimentation. Can mesenchymal stem cells have the ability to differentiate into certain tissues and develop the corresponding functions after they are transplanted into certain tissues?OBJECTIVE: To observe the differentiation of the bone marrow mesenchymal stem cells into the nerve and its effect on the nerve functional recovery after they are transplanted into the peripheral zone of the ischemic infarction focus of the cerebral cortex.DESIGN: A randomized controlled study.SETTING: The Department of Anatomy of the School of Basic Medical College of Sun Yat-sen University; the Department of Neurology of the First Hospital of Sun Yat-sen University; Department of Pathology of the Medical College of Jinan University.MATERIALS: The experiment was conducted at the Medical College of Sun Yat-sen University from November 2002 to November 2003. Forty-eight male SD rats were chosen and randomly divided into two groups, with 32 rats in cerebral infarction model group and 16 in the non-model control group. In the cerebral infarction group, the rats were randomly divided again into two groups: 16 rats in the transplantation group and 16 in the phosphate buffered fluid group. The anterior fontanel taken as the reference point, 5 μL(5 × 104 L-1) of the bone marrow mesenchymal stem cells or the phosphate buffered fluid was respectively transplanted at the site 3 mm away on the caudal side and 1.5 mm aside at the depth of 2. 0 - 3. 0 mm.METHODS: The bone marrow mesenchymal stem cells were obtained through the separation and purification of the bone marrow of the ribs taken away in the thoracic surgery from the patients without the hematological diseases, and then the cells underwent in vitro culture, the amplification and the identification. At the 2nd and 6t1 weekend after the transplantation,the rats of every group were anesthetized, and the samples were taken from the transplantation site and made into the 25 μm of continuous frozen section. Then, the immunohistochemical method was used for the detection of the expressions of neuron specific enolase, neurofibril protein, glial fibrillary acidic protein, and nidogen to evaluate the ability of the bone marrow mesenchymal stem cells to differentiate into neurons and glial cells. Eight rats of the transplantation group and 8 rats of the phosphate buffered fluid group were randomly taken out, and 2 and 6 weeks before and after the transplantation the bar walking test evaluation method was used to identify the general status and reaction ability of the rats. Sixteen rats of the control group were assessed at the same time.enolase, neurofibril protein, glial fibrillary acidic protein and nidogen in the bar walking test.2nd weekend after the transplantation, there were positive expressions of glial fibrillary acidic protein and nidogen at the transplantation site of the bone marrow mesenchymal stem cells. At the 6th weekend there were positive expressions of neuron specific enolase and neurofibril protein at the transplantation site of the bone marrow mesenchymal stem cells. While in the phosphate buffered fluid group, there were negative expressions of neuron specific enolase, neurofibril protein, glial fibrillary acidic protein and nidosymptoms in the control group and the evaluation scores were all 9. 2 weeks after the transplantation, and the evaluation scores of the motor function in the transplantation group were higher than the ones in the phosphate buffered fluid group, [(6.7±0.9), (5.3-±1.0), (P ＜0.05)]. Six weeks after the transplantation, the evaluation scores of the motor function in the transplantation group were also higher than those in the phosphate buffered fluid group[(8.9±1. 1),(7.2±0.8),(P ＜0.05)].CONCLUSION: After their transplantation into the central nervous system,the bone marrow mesenchymal stem cells showed the ability to differentiate into neurons and glial cells, in which the characteristics of some neurons and glial cells were found. Bar walking test found that the evaluation scores of the motor function in the transplantation group were higher than those in the phosphate buffered fluid group, which suggests that the bone marrow mesenchymal stem cells have a significant effect on restoration of the functions of the nerves.

AIM: To observe the effect of adrenocorticotropic hormone(ACTH) on the levels of brain-derived neurotrophic factor (BDNF), trkB and corticotropin-releasing hormone(CRH) in the hippocampus of arthritic rats.METHODS: The BDNF immunoreactivity (IR) and CRH-positive neurons were stained with immunohistochemistry and in situ hybridization methods, respectively.RESULTS: The BDNF-IR, CRH mRNA-positive neurons in the contralateral hippocampus of the arthritic rats were increased significantly, which was decreased markedly by intraperitoneal injection of ACTH. However, the effect of ACTH was attenuated after adrenalectomy (ADX).CONCLUSION: These results suggest that BDNF and CRH in the hippocampus of arthritic rats were involved in the modulation of chronic pain, ACTH produced its analgesic effect by inhibiting the increase in BDNF and CRF level. Adrenal is critical to the analgesic action of ACTH.