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Objective To investigate the effects of ferulic acid (FA) and adipose-derived mesenchymal stem cells(ADMSCs)on rat hepatic stellate cells (HSCs) by regulating TGF-β/smad signal transduction pathway.Methods HSCs were divided into 4 groups:blank control group,FA control group,ADMSCs control group and FA+ADMSCs group.The apoptosis rate of HSCs in each group was detected by flow cytometry.The expression levels of TGF-β1,Smad2,Smad3 and Smad7 mRNA in HSCs were detected by qRT-PCR.The TGF-β1 and Smad7 protein levels,and phosphorylated-Smad2/3 (p-Smad2/3)expression were detected by Western blot.Results Compared with the other 3 groups,the apoptosis rate of HSCs in the FA+ADMSCs group was significantly increased(P＜0.05),while the expression levels of TGF-β1,Smad2 and Smad3 mRNA were significantly decreased,and the Smad7 mRNA expression was increased;moreover,the expression levels of TGF-β1 protein and p-Smad2/3 were significantly decreased,while the Smad7 protein expression was significantly increased(P＜0.05).Conclusion FA can enhance the effect of ADMSCs for down-regulating the TGF-β1 expression in HSCs,and then leads to the decrease of downstream p-Smad2/3 activity and Smad7 expression increase,thus participates in promoting cellular apoptosis.
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Objective To investigate the clinical characteristics of elderly-onset inflammatory bowel disease (IBD) and its treatment options.Methods Clinical data of IBD in-patients treated in Peking University First Hospital from January 1995 to December 2014 were reviewed and the subsequent information in the follow-up was also collected.Based on the onset age, patients were divided into the elderly-onset group (≥60 years old) and the control group (＜60 years old).The general situation, clinical characteristics and treatment options were retrospectively analyzed and compared between the two groups.Results There were 37 patients (9.3％) with IBD in the elderly-onset group, with 25 cases with ulcerative colitis (UC) and 12 cases with Crohn disease (CD).The ratio of gender, smoking, family history of IBD and colon cancer in the elderly-onset and control groups were 1 : 0.9 and 1 : 1.3, 32.4％ and 21.7％, 0.0％ and 6.1％, 0.0％ and 1.9％, respectively, with no significant differences between the two group (P=0.597, 0.139, 0.247 and 0.840).The attack frequency and the percentages of patients with persistent symptoms and severe disease had no statistical differences between the elderly onset and the control groups.Compared with the control group, the elderly-onset group showed that the percentage of UC patients with extensive colitis (E3) was decreased [16.0％ (4/25) vs.41.2％ (117/289) respectively (x2 =6.123, P=0.013)], and the percentage of CD patients with colonic type (L2) was increased [58.3％ (7/12)vs.21.3％ (16/75) respectively (x2=6.447, P=0.011)].For treatment options, the number of patients receiving 5-Aminosalicylic Acid (5-ASA)/sulfasalazine(SASP) as maintenance treatment for more than 1 years were 274 cases (96.9％) in the control group and 22 cases (86.5％) in the elderlyonset group (x2 =6.382, P=0.011).And the number of patients receiving systemic corticosteroids during induction period were 5 cases (19.4％) in the elderly-onset group and 122 case (43.2％) in the control group (x2 =7.617, P=0.006).There were no statistically significances in the number of patients with immunosuppressant, biologics and operation treatment between the two groups.Conclusions The elderly-onset IBD patients are not rare.The inflammation involvement is more limited in the elderly-onset patients than in the young-onset patients.The treatment option for elderlyonset IBD should be more cautious, especially for the treatment with 5-ASA/SASP long-term maintenance and systemic corticosteroids usage.
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BACKGROUND:Previously, serious thoracolumbar burst fractures with spinal cord injury were mainly treated by anterior surgery. The superiority of conventional posterior repair lies in strong reduction and fixation effects. However, it is difficult to fuly decompression. The anterior and central cylinders of the injured vertebra cannot receive good reconstruction. Loss of correction and internal fixation failure always appear. It remains controversial which repair method is more ideal. OBJECTIVE:To comparatively analyze the effect of posterior and anterior approaches with subtotal vertebrectomy, decompression, reconstruction of spine, and internal fixation for patients with thoracolumbar burst fractures. METHODS: The data of 42 patients with thoracolumbar burst fracture treated by anterior and posterior approaches with subtotal vertebrectomy, decompression, and reconstruction of spine from May 2006 to December 2012 was retrospectively analyzed. They were divided into two groups according to the surgical procedures: anterior approach group (n=23) and posterior approach group (n=19). They were folowed up for at least 24 months. Repair results and imaging results were compared in patients of both groups. RESULTS AND CONCLUSION:None patients in the two groups affected fixator loosing or breakage, and obtained good bone fusion. The intraoperative blood loss was more and operative time was longer in the anterior approach group than in the posterior approach group, showing significant difference (P 0.05). No significant difference in the anterior vertebral height and kyphosis Cobb angle was detected before treatment, at 1 week after treatment and during final folow-up (P > 0.05). The incidence of postoperative complications was significantly higher in the anterior approach group than in the posterior approach group (P < 0.05). These findings confirm that subtotal vertebrectomy through anterior approach and posterior approach can effectively repair thoracolumbar burst fractures. However, the complications of posterior approach are less, and posterior approach has few impacts on the pulmonary function, is safe, and has good biocompatibility with the host.
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Objective To explore the clinical effects of pyloroplasty on proximal gastrectomy in patients with lesions in cardia of the bottom of stomach.Methods One hundred cases of the proximal gastric diseases who were enrolled in the second affiliated hos-pital of Chongqing medical university from the January 2010 to December 2013 were selected in this prospective randomized con-trolled experimental study.Among them,6 cases were excluded,and the other 94 cases were divided into two groups:the experimen-tal group (A),in which 54 patients were underwent additional pyloroplasty surgery;the control group (B),in which 40 patients were underwent with no strings attached pyloroplasty.All patients were followed up.Then,the clinical indicators during hospitaliza-tion,term complications and repeated endoscopy prompted pyloric stenosis and reflux esophagitis situation three months after dis-charged from hospital were observed.Results There was no significant difference in the incidence rate of gastroplegia (P >0.05);there were significant differences in other clinical indicators during the postoperative hospitalization and the incidence rate of compli-cations (P <0.05).Conclusion Proximal gastrectomy additional pyloroplasty could reduce gastric retention,decrease the incidence of reflux esophagitis,shorten the hospital period and could be helpful for recovery,thus it is recommended to be further promoted.
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Objective To observe the effect of glutamine-enriched early enteral nutrition on intestinal mucosal barrier injury after orthotopic liver transplantation in rats.Method Male Wistar rats,the recipients were randomly divided into three groups:control group (control group,n =10),orthotopic liver transplantation group (OLT group,n =30) and glutamine-enriched early enteral nutrition group (EEN group,n =30).Only dissecting hepatoduodenal ligament was performed in control group,and OLT was performed from Wistar to Wistar rats by modified two-cuff method in OLT group and EEN group.For EEN group,recipients were supplied with Nutrison Fiber (125 ml/kg every day) plus Gln (0.3 g/kg every day) for 3 days before and 3 h after surgery by gastric perfusion.For OLT group,the same volume of normal saline was administered instead of the Nutrison in the same time.No special treatment was given in control group.The levels of plasma endotoxin,D-lactic acid,and TNF-α were determined at different time points in the three groups.The ultrastructural changes of ileal mucosa were observed under the transmission electron microscopy.At the same time,the remaining 5 rats per group were used for observing the lifetime.Result As compared with control group at 12,24 and 72 h,the levels of plasma endotoxin,D-lactic acid,and TNF-α were significantly increased in OLT group and EEN group (P＜0.01),and as compared with OLT group,the above-mentioned indexes were obviously decreased in EEN group at 24 and 72 h (P＜ 0.01).The ileal mucosal epithelial clearance in control was normal,and microvilli arranged neatly under the electron microscope.the ultrastructure damage in OLT group was more serious than in EEN group at 12,24,and 72 h.As compared with control group at 12,24,and 72 h,the expression levels of TNF-α mRNA were significantly increased in OLT group and EEN group (P＜0.01),and as compared with OLT group,the expression levels of TNF-α mRNA were obviously decreased in EEN group (P＜0.01).The survival time in cntrol group was significantly longer than in OLT group and EEN group (P＜0.05),and as compared with OLT group,the survival time in EEN group was obviously extended (P＜0.05).Conclusion OLT can lead to the damage of intestinal mucosal barrier,and glutamine-enriched early enteral nutrition is a potent protection against intestinal mucosal barrier injury and prolongs the survival time of rats after OLT.
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Objective To investigate the feasibility of a rat carotid atherosclerosis model induced by artery clamping with high-cholesterol diet and vitamin D3.Methods Twenty Wista rats were randomly divided into either a general diet group (n =5) or a high cholesterol diet group (n =15).After clamping the right common carotid arteries of the rats in the high cholesterol diet group,they were fed with high fat diet,and vitamin D3 (600 000 IU/kg) was injected intraperitoneally.The lipid levels of the general diet group and the high cholesterol diet group were detected at 6 and 12 weeks respectively.The bilateral cormmon carotid arteries were selected for preparing paraffin sections and were stained with HE staining.The pathological changes in blood vessels were observed.Results The levels of serum total cholesterol at 12 weeks (2.803 ± 1.307 mmol/L vs.1.513 ±0.281 mmol/L; P=0.017) and low-density lipoprotein cholesterol (0.660± 0.260 mmol/L vs.0.311 ±0.078 mmol/L; P =0.003) in the high-cholesterol diet group were significantly higher than those at 6 weeks.The histopathological examination showed that the common carotid artery intimas on the clamping sides were incomplete,the foam cell deposition was observed under intima,the atherosclerotic plaques or fibrous plaques were observed on the surface of cavity,inside the plaques contained necrotic tissue,and thrombosis was observed in the cavity.The common carotid artery intima-media thickness in the general diet group (n =5) at 12 weeks was 8.3 ± 1.1 μm.The sham-operated sides (n =20) and clamping sides (n =20) were 8.8 ± 0.7 μm and 97.4 ±25.7 μm,respectively.There were significant differences among the three groups (F =116.313,P=0.000).The clamping sides in the high-cholesterol diet group were significantly higher than the shamoperated sides in the high-cholesterol diet group (P=0.000) and the general diet group (P =0.000).Conclusions Common carotid artery clamping with high-cholesterol diet and vitamin D3 is a simple and feasible method for inducing a rat carotid atherosclerosis model.
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BACKGROUND: The implantation technique of bone marrow mesenchymal stem cells (MSCs) is of important significance for repair of brain injury. However, its action pathway still needs to be investigated.OBJECTIVE: To observe the changes of Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus of rats with cerebral ischemia in which MSCs were implanted, and to analyze the action mechanism of intracranial implantation of MSCs inhibiting neuronal apoptosis.DESIGN: A randomized controlled experiment.SETTING: Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-four male adult Wistar rats, weighing 180 to 240 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomized into 3 groups with 8 in each: control group, injury group and implantation group. 5-bromodeoxyuridine (BrdU, Sigma Company), TUNEL kit, Bcl-2, Bax antibody kit were purchased from Beijing Zhongshan Bioengineering Company.METHODS: This study was carried out in the Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College between April 2005 and September 2006. Rats in the injury group and implantation group were developed into rat models of cerebral ischemia by suture of external carotid artery. Seven days later, the successful rat models in the implantation group were injected in the cerebral cortex and striatum with 2×1012 L-1 MSCs suspension primarily cultured in vitro. The processing of the experimental animals corresponded to the requests of Animal Ethics.MAIN OUTCOME MEASURES: Neuronal apoptosis and Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus were detected by TUNEL staining and immunohistochemical method on the 7 and 14 days after successful modeling, separately.RESULTS: ①Neuronal apoptosis: On the 7th day after successful modeling, apoptotic cells were not found in the control group, and apoptotic cells in the implantation group were significantly fewer than those in the injury group (P ＜0.01). ② Bcl-2 and Bax expressions: On the 14th day after successful modeling, Bcl-2-positive neuronal expression in the cerebral cortex and hippocampus of rats in the injury group was significantly weaker than that in the control group and implantation group (P ＜ 0.01). Bax-positive expression in the cerebral cortex and hippocampus of rats in the injury group was significantly stronger than that in the control group and implantation group (P ＜ 0.01).CONCLUSION: MSCs can promote Bcl-2 expression and inhibit Bax expression of rats with cerebral ischemia injury,and accordingly neuronal apoptosis will be reduced.
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BACKGROUND: The c-fos gene is commonly expressed in neurons,which may act as one of the signs of activities and reflection of injured neural cells.OBJECTIVE: To investigate the expression of hippocampal c-fos gene of rats with ischemic cerebral injury and the protective role of shenmai injection at the molecular level.DESIGN: Randomized controlled study.SETTING: Department of Histology and Embryology, Luzhou Medical College.MATERIALS: The experiment was completed at the Department of Histology and Embryology, Luzhou Medical College from January to March 2002. Totally 30 male Wistar rats were randomly divided into control group, ischemia group and treatment group with 10 in each group.METHODS: Bilateral common carotid arteries of rats in the ischemia and treatment groups were splinted for 30 minutes and reperfused for 1 hour to establish models of ischemic cerebral injury. Rats in the treatment group were injected with 2 mL/kg of shenmai (constituted with hongshen, dwarf lilyturf tuber and other Chinese herbal medicines) 30 minutes before ischemia. Rats in the ischemia group were not treated with any drugs, and rats in the control group were treated with sham operation but without splinting common carotid artery and giving any drugs. Hippocampal tissue of rats was obtained to make paraffin sections. In every group, one section from each rat was taken at random. Totally 100 neurons of every group were counted. Expression of c-fos gene in hippocampal neuron was observed according to the nuclear color of neurons (+++ as dark-brown mark;++ as brown mark; + as light brown mark; - as no brown mark).MAIN OUTCOME MEASURE: c-fos expression of hippocampal neurons of rats in each group.RESULTS: Totally 30 rats entered the final analysis. Expression of c-fos gene in hippocampal neurons was significantly more in the ischemia group than that in the control group (+++: 24/visions, 7/visions, P ＜ 0.05); but that in the treatment group was less than that in the ischemia group (+++:13/visions, 24/visions, P ＜ 0.05).CONCLUSION: Shenmai injection can reduce the expression of c-fos gene in hippocampal neurons of rats with ischemic cerebral injury, and can protect neural cells of ischemic cerebral injured tissue.
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Objective To study the apoptosis of endothelial cells and the relation between it and the expression of P53 protein after focal cerebral ischemia reperfusion in rats.Methods The apoptosis of endothelial cell 2,6,12 h and 1,2,3,7,14,21 d after cerebral ischemic reperfusion were observed using an in situ end labeling method,and the expression of P53 protein was detected by immunohistochemical staining methods.Results Apoptotic endothelial cells in ischemic penumbra were observed 2h after cerebral ischemic reperfusion, they peaked at 12~24 h, and decreased gradually.There was no remarkable difference between it and the sham operative group at 21 d.The P53 protein began to express 6h after cerebral ischemic reperfusion, it peaked at 1~2 d, and then declined gradually to controlled level at 7d. The expression peak time of P53 was 24 h later than that of cell apoptosis.Conclusion Apoptosis was a pattern of endothelial cell death after reperfusion of MCAO. P53 protein played an important role in the process of apoptosis of endothelial cells.
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0.05).The expression began to increase around the infarction at 12h reperfusion,it reached the peak at the 3th day in the boundary zone and then it gradually decreased.Conclusion The overexpression of IL 6 played an important role in the course of cerebral ischemic reperfusion, and could damage the brain tissue.