BACKGROUND: Preliminary study has proved that the bone marrow-derived mesenchymal stem cells (MSCs) in a rat emphysema model produced by use of trypsin alone can "homing" to the lesioned lung tissues, and participate in the formation of pulmonary arteries to promote lung tissue repair. Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) play equally a powerful role in promoting angiogenesis. OBJECTIVE: To observe the influence of bFGF, VEGF and MSCs in regeneration of pulmonary capillary and pathological repair of pulmonary emphysema rats. METHODS: Except normal control group, the remaining 5 groups of rats were exposed to tobacco smoke and received a single intratracheally instillation of porcine pancreatic elastase to induce emphysema models. Following successful modeling, rats of bFGF group were intratracheally injected with 400 U bFGF and rats of VEGF group with 2 μg VEGF, once a week for three times. MSCs group was injected 1 mL suspension of 4×10~9/L MSCs into tail vein. MSCs+VEGF group was injected MSCs into tail vein and intratracheally injected VEGF (2 ug, three times) at the same time. Model control and normal control groups were intratracheally injected with equal volume of sodium chloride. Four weeks after treatment, arterial blood gas analysis was performed to observe pathological and morphological changes of lung tissues. CD34~+ expression in lung tissues was determined using immunohistochemistry method. RESULTS AND CONCLUSION: Compared with model control group, PaO_2 values dramatically increased in VEGF group (P <0.05), while other indices remained unchanged (P > 0.05); there were no obvious changes in each index in other groups (P >0.05). Gross and microscopic observations showed that, lung was smooth, pale pink, and elastic in normal control group, with uniform size of pulmonary alveoli on cross-section; pathological changes of chronic obstructive pulmonary emphysema existed in model control group, but improved in other 4 groups. Compared with model control group, mean pulmonary alveoli number and CD34~+ relative positive area dramatically increased in bFGF, VEGF, MSCs, MSCs+VEGF groups (P < 0.05), mean linear intercept and mean alveoli area were significantly reduced (P < 0.05). No significant difference was observed in each index among these 4 groups (P > 0.05). bFGF, VEGF and MSCs could improved the pathology of pulmonary emphysema models produced by tobacco smoking and intratracheally instillation of porcine pancreatic elastase. The possible mechanism of recovering the pulmonary emphysema is the proliferation of pulmonary capillary and enlargement of pulmonary artery, improved blood flow in the lung, improved ventilation/perfusion shunt, reduced pulmonary alveolus size and volume of the lung through self-compensation.

Objective To investigate the clinical manifestations,endoscopy and pathological features of Behcet's disease (BD) in gastrointestinal systems in elderly patients.Methods Totally 8 patients aged ≥ 60 years with BD were collected.Clinical characteristics and endoscopic performances of gastrointestinal tracts in patients were summarized.All patients were received gastrointestinal biopsy and the histology change was observed.The vasculitis of gastrointestinal tracts was confirmed by histochemical staining.Results The proportion of patients with BD involving the esophagus was 16.7％ (9/54) and among whom 6 cases (11.1 ％) were elderly,so the proportion of BD patients with esophageal lesions in elderly patients was 66.7 ％ (6/9).The proportion of patients with BD involving the ileocolon was 9.3％ (5/54) and 4 cases (7.4％) were elderly patients,so the proportion of BD patients with ileocolon lesions in elderly patients was 80.0％ (4/5).6 patients had retrosternal chest pain,among whom 2 patients with dysphagia and 4 patients with abdominal pain and hematochezia.In the laboratory tests,the levels of erythrocyte sedimentation rate (ESR) and C reactive protein were elevated in 4 cases,and antinuclear antibodies (ANA) and anti-neutrophil cytoplasm antibody(ANCA)were negative in all patients.In the physical examination,oral and genital ulcers in 6 patients,and a history of recurrent oral and genital ulcers were found in the other 2 patients.6 patients had ophthalmia and 4 patients with skin lesions.6 patients had multiple esophageal ulcers,in which shallow ulcers was more common,and the surface of ulcer was clean and white with congestion and edema at the periphery of ulcers.BD involving the ileocecal and ascending colon were found in 4 cases.Typical small vessel vasculitis was the pathological performance.The retrosternal chest pain disappeared and the esophageal ulcers were healed after the treatment.Conclusions Gastrointestinal involvement in BD patients is associated with the the course of disease.Longer duration can increase the possibility of gastrointestinal involvement in BD.Elderly BD patients are more common with esophageal involvement.Rctrosternal chest pain may be a relatively specific clinical symptom in BD involving esophageal tract.Glucocorticoids and (or) thalidomide are effective for the treatment of BD with esophageal involvement.

BACKGROUND: Pathological changes of pulmonary emphysema are not reversible according to the existent pathogenesis of pulmonary emphysema. Research over many years report that injury of pulmonary blood capillary may take part in new pathogenesis of pulmonary emphysema based on lung volume reduction operation and bronchial lumen occlusion. Mesenchymal stem cells (MSCs) have multi-directional differentiation potencies, such as the differentiation into vascular endothelial cells. Therefore, MSCs may promote pulmonary vascularization and repair pulmonary tissue.OBJECTIVE: To observe the effect of MSCs transplantation on pathological changes of arterial blood gas and pulmonary tissue in model rats with pulmonary emphysema, and investigate the therapeutic effects on MSCs on pulmonary emphysema and the pathogenesis of pulmonary emphysema.DESIGN: Randomized controlled animal study.SETTING: The Second Hospital of Shanxi Medical University.MATERIALS: Thirty healthy Wistar rats, 6 weeks old, of either gender, weighing 180-200 g. They were provided by Physiological Experiment Animal Center, Shanxi Medical University. All rats were randomly divided into MSCs treatment group, model group and control group with 10 rats each.METHODS: The experiment was carried out in the Physiological Laboratory of Shanxi Medical University from April 2005 to April 2006. Rats in the MSCs treatment group and in the model group were anesthetized and intratracheally perfused with 250 U/kg Porcine pancreatic elastase (PPE) to establish pulmonary emphysema models; while, rats in the control group were perfused with saline. The models were successfully established 4 weeks later. All rats were anesthetized and then femur and tibia were obtained to separate and culture MSCs in vitro. Immunocytochemistry was used to detect the expression of CD71 in order to evaluate MSCs. Bromium azacytidine-labeled MSCs were inserted along caudal vein into rats in the MSCs treatment group; while, rats in the model group and control group were inserted with the same volume of PBS solution.MAIN OUTCOME MEASURES: ① Changes of arterial blood gas in the three groups; ② Pulmonary tissue was used for pathological sections in order to calculate mean alveolar number, mean alveolar area and mean linear intercept; ③Immunocytochemical staining was used to measure numbers of CD34+ cells so as to determine proliferation of alveolar blood capillary.RESULTS: Three rats in all died during the model establishment, while another 3 rats were supplied. Therefore, an overall number of 30 rats were involved in the final analysis. ① Culture and evaluation of MSCs: At 3 days after inoculation, MSCs were generally adherent to walls and fusiformly shaped. In the third generation, the expression of CD71 was observed on the surface of MSCs.② Comparisons of arterial blood gas in the three groups: There were no significant differences in pH value, PO2, PCO2 and SaO2 in the three groups (P ＞ 0.05). ③ Pathological changes of pulmonary tissue: Pathological changes in the MSCs treatment group were milder than those in the model group;meanwhile, mean alveolar number in the MSCs treatment group was more than that in the model group, and there was significant difference between them (F=80.201, P＜ 0.05). While mean alveolar area and mean linear intercept in the MSCs treatment group were smaller than those in the model group, and there were significant differences (F =26.755,26.875, P ＜ 0.05). ④ Comparisons of CD34+ expression in pulmonary tissue: Relative positive area of CD34+ in the MSCs treatment group and model group was smaller than that in the control group (F =20.411, P ＜ 0.05), but that in the MSCs treatment group was larger than that in the model group, and there was significant difference between them (F=20.411, P＜ 0.05).CONCLUSION: MSCs can reverse the pathological changes of pulmonary emphysema; on the other hand, the decrease of the number of pulmonary capillary maybe one of the important pathogeneses of pulmonary emphysema.

Translational medicine is a more effective and efficient way to improve the investment return on bioscience and provide equitable access to high—quality healthcare.Recognizing the need for a new impetus to spur clinical and translational research,the National Institutes of Health established the Clinical and Translational Science Awards Program.

Objective To explore the possible mechanism of SchA ,which decreases MPP+induce SH-SY5Y cell damage .Meth-ods Cultured cells were divided into 5 groups ,one as control group ,cultured by free-blood serum media;the other 4 groups were treated with different concentrations of SchA(1 ,3 ,5 μmol/L) and MPP+ (1 mmol/L) for 48 h named model group ,1 ,3 ,5 μmol/L SchA group respetivly .The content of nitric oxide(NO) were measured by NO kit ;The expression levels of total Akt and p-Akt proteins were detected by Western blot .Results Compared with the control group ,the content of NO in group significantly in-creased after MPP+stimulating(P<0 .05);compared to the control group ,the content of NO in 5μmol/L SchA group significantly decreased(P<0 .05) .The expression levels of total Akt in all groups had no significant difference(P>0 .05) .The expression levels of p-Akt in model group significantly lowered ,while SchA(1、3、5 μmol/L) significantly increased the expression levels of p-Akt in comparision with cells in model group .Conclusion Decreasing MPP+ induced SH-SY5Y cell damage of SchA may be related to the content of NO and p-Akt expression .

BACKGROUND:Recently,application of stem cells and growth factor to promoting lung regeneration in repair of emphysema lesion has been a hot focus in study.Thus,it is worth to pay attention on whether stem cells carrying relevant foreign growth factor gene can repair emphysema lesion.OBJECTIVE:To evaluate the effidency of adenovirus vector mediated green fluorescence protein(Ad-GFP)transfecting bone marrow mesenchymal stem cells(BMSCs)and its effect on the cell proliferation,to explore oriented migration of intravenously administrated BMSCs transfected with Ad-GFP in the lung tissues of pulmonary emphysema rats.METHODS:MSCs were separated and purified from the bone marrow of rats by density gradient centrifugation and by adherence.At different multiplicity of infection(MOI),transfection efficiency was observed by laser confocal microscopy.At 48 hours of transfection,MTT method was used to evaluate the proliferation of MSCs.A total of 16 Wistar rats were randomly divided into emphysema model group and control group(n=8).Model rats were established by exposure to cigarette smoke.MSCs,transfected with Ad-GFP,were grafted into the body of rats via tail vein.Lungs derived at 24 hours after implantation,and frozen sections were made.Migration and survival of MSCs in the lung tissues were observed by fluorescence microscopy.RESULTS AND CONCLUSION:MSCs from Wistar rats were successfully cultured,grew well and infected by Ad-GFP.The highest transfection effincincy(88.42 %)could be achieved at MOI of 200.Green fluorescent protein labeling had little effect on proliferation of MSCs by different MOI(P＞0.05).At 24 hours posttransplantation,the green fluorescence-positive tissue was Found in the lung tissues of emphysema model group and control group.Compared with control group,the expression of GFP in lung tissues was higher in emphysema model group(P＜0.05).These suggested that introduction of target gene cannot affect proliferation and homing property of BMSCs.

BACKGROUND:The pathogenesis of intervertebral disc protrusion remains unclear.A stable and feasible intervertebral disc degeneration model is required to validate the pathology and imaging performance and to reveal intemal factors.OBJECTIVE:To investigate the feasibility of building rabbit model of intervertebral disc degeneration using aspiration method,as well as the convenience and stability of model establishment through the performance of pathology and imaging.MATERlALS:A total of 4210-month-old healthy New Zealand rabbits,irrespective of genders,were randomly divided into control group and experiment group with 21 rabbits in each group.Sumianxin was offered by Veterinary Institute,Quartermaster University of Chinese PLA in Changchun City.METHODS:The disc height index(DHI)of all rabbit swas set as 100%.A21-gauge hypodermic needle was used to aspirate nucleus pulposus 8-12 mg from L4-5 disc of rabbits in the experiment group,while control group received no management. At 4,12.20 weeks,7 rabbits were selected from each group for index determination.MAIN OUTCOME MEASURES:DHI percent,X-ray plain film,MRI and Alcian blue staining were used to observe the changes in the intervertebral disc degeneration.RESULTS:The DHI percentage of experiment group at 4,12 and 20 weeks respectively was(81.0±3.2)%,(75.0±2.5)%and (71.0±1.8)%,with significant difference compared to control group(P＜0.05).T2-weighted image showed that the signal of the discs was significantly decreased,and Alcian blue staining showed that the content of aggrecan was also significantly decreased.CONCLUSI0N:The aspiration is a reliable and convenient way to construct rabbit models of intervertebral disc degeneration.and these models exhibit the similar pathology and imaging performance with human intervertebral disc degeneration.

Objective To evaluate the clinical efficacy and safety of hydrotalcite chewable tablets in treatment of patients with functional dyspepsia epigastric pain syndrome(EPS), and to investigate the onset time of hydrotalcite after the first dosage and whether use of hydrotalcite in EPS is a costeffective strategy. Methods A multicenter, randomized, open, positive controlled clinical trial was carried out in 240 patients with EPS. The subjects randomly received eigher hydrotalcite or omeprazole for 2 weeks. The improvemcnt and the disappear time of symptoms were evaluated before and after treatment and cost-effective was analyzed between two groups. Results It was demonstrated that both hydrotalcite and omeprazole could relief symptoms after treatment. After treatment for 2 weeks, the total effective rate was 85. 71% in hydrotalcite group and 90. 43% in omeprazole group with no significant difference (P＞0.05). The median onset time of hydrotalcite after first dosage was 0.417 h. The overall direct cost per patient was RMB ￥122. 29 for hydrotalcite treatment and RMB ￥242.95 for omeprazole treatment. The main adverse events included diarrhea, thirst, bloating,headache and belching. No severe adverse event was found in two groups. Conclusions Hydrotalcite has fast effect on relief of EPS symptoms. Use of hydrotalcite is a cost-effective strategy in the management of EPS. It is a safe and effective medicine in treatment of EPS.

Objective To study the influence of total flavonoids of hemerocallis fulva(TFHF) on hepatocyte apoptosis and related protein expression in mice with alcoholic hepatic injury.Methods A total of 40 mice were randomly divided into four groups:blank control,model control andsmall and high dose TFHF groups,10 cases in each group.The mice were given the continuous gavage administration for 7 d.Then the model group was given once gavage by 50％ ethanol 12.0 mL/kg after 1 h of the last administration.The blank control group was given the equal volume of distilled water.The activity levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum as well as the superoxide dismutase(SOD) activity and malondialdehyde (MDA) content in liver tissue hemogenate were detected.Hematoxylin and Eosin(HE) staining was performed for observing the pathological changes of the liver tissue.The flow cytometer was used to test the apoptosis ratio in hepatocyte suspension.The expressions of caspase-3,Bcl-2 and Bax protein were detected by Western blot.Results The various TFHF groups could decrease the activities of ALT and AST in serum (P＜0.05),while could decrease the MDA content in liver tissue hemogenate (P＜0.01) and increased the SOD activity;the liver tissue pathological examination showed that the high dose TFHF group could make the liver cell degeneration,alleviated the necrosis degree and relieved the pathological change of hepatic tissue;compared with the model group,the hepatocyte apoptosis rate in each TFHF group was decreased significantly;Western blotting results showed that the caspase-3 protein level in each TFHF group was decreased,expression of Bcl-2 protein was increased,whereas which of Bax protein was decreased and Bax/Bcl-2 ratio was reduced.Conclnsion TFHF has obvious protective effect on mice acute hepatic injury induced by ethanol,and can inhibit the hepatocyte apptosis,its action mechanism may be related to its antioxidation and regulation of caspase-3,Bcl-2 and Bax expression.

The cancer stem cells (CSCs) from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified. The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension, and mixed homogeneously into 1.2% alginate gel. Single-cell alginate gel was cultured with serum-free DMEM/F12 medium. Epirubicin (0.8 mug/mL) was added to the medium to enrich CSCs. After cultured conventionally for 7 to 10 days, most of cells suspended in alginate gel were killed by epirubicin. But few cells survived and some single-cell cloning spheres formed. Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation. Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo. The results showed that some cells positive for Oct3/4 (TRITC) and Nanog (TRITC) were found in single-cell cloning spheres, and most of positive cells were concentrated in the core of sphere. Cells from spheres could form osteosarcoma in the body of mice. It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes (Oct3/4 and Nanog), resisting anti-cancer drugs, and tumorigenicity in vivo. To sum up, it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.