Objective To observe the bacterioflora of bile and it' s drugs sensitivity in patients with bile duct diseases to serve as a guidance in medication. Methods Bile of 156 cases of patients with biliary disease was collected and cultured respectively during endoscopic retrograde cholangiopancreatography (ER-CP) by deep cannulation and sucking out bile through the catheter. Forty eight blood samples among them were cultured simultaneously. Ten kinds of drug papers were used to investigate the bacterial sensitivity. The data were analysed statistically- Results Bacteria positive rate of these 156 bile samples was 62. 8% , including Gram - negative bacteria (81.6% ) and Gram - positive bacteria (18. 4% ). These bacteria consist of Pseudomonas aeruginosa (35. 1% ) , Escherichia coli(22. 8% ) , Streptococcus faecalis (16.7%) ,Entero bacilliaerogenes(7. 9% ) ,Klebsiella(7. 0% ) , Citrobacter(6. 1% ) ,Acinetobacler lwqffi(2. 6% ) and Staphylo-coccus aureus( 1. 7% ). The bacteria positive rate was only 4. 2% in the 48 blood samples. Drug sensitive rates of bacteria against 10 kinds of drugs were changed in recent years. The result indicated that the sensitive rates of bacteria were significantly higher in ciprofloxacin, fortum, cefoperazone, sulbactam + cefoperazone and cilastatin than those in ampicillin,azlocillin, cefazolin sodium, eefuroxime(P

Objective To study the clinical effects of human amniotic membrane transplantation loaded with autologons marrow mesenchymal stem cells(MSCs) in treatment of pterygium patient. Methods Totally 10 patient (10 eyes) with primary giant pterygium aged 42 ～60 years(7 males and 3 females) were enrolled. All patients were informed consents. Bone marrow was sterilely collected by bone marrow aspiration. MSCs were harvested, purified and cultured. At the third passage,MSCs were incubated in amniotic membrane. After pterygium resection,amniotic membrane patch with cells surface upward was covered on surface of exposed sclera. Another simple amniotic membrane was sutured to protect transplanted cells. Results 1 ～ 3 days after dissection of corneal epithelial area was repaired,a week around later,migration of conjunctival epithelial gradually grew,about 10 ～ 14 days, absorption and fusion of amniotic membrane began. Follow-up of 6 months to 3 years, there was no recurrence. Conclusion After amniotic membrane loaded with MSCs were transplanted to the ocular surface ,corneal epithelium repaired rapidly ,transplanted amniotic membrane and conjunctival integrated, conjunctival irritation was few, ocular surface scar was light, the pterygium was no recurrence.

Fast track surgery(Frs) is an idea and theory which has been highlighted by more and more clinic doctors and scholars, and widely used in the surgical operations recently. ~TS has greatly changed the mode of clinic therapy, ameliorated the velocity of the rehabilitation and prognosis after operation, and improved the quali- ty of life and the therapeutic effect. We reviewed the application of FTS in the surgery in this article.

Endocardial Cushion Defects ; complications ; Female ; Humans ; Infant, Newborn ; Lung ; abnormalities ; Lung Diseases ; complications ; congenital

Objective To investigate the effect of Mycobacterium vaccae vaccine combined with chemotherapy on immune function in treatment of children with pulmonary tuberculosis.Methods 89 cases with pulmonary tuberculosis from December 2010 to December 2013 treated in three hospitals in xinxiang city were randomly divided into control group and observation group.The control group was given regular chemotherapy,the observation group was given chemotherapy combined with Mycobacterium vaccae vaccine therapy.CD4 +,CD8 +,CD8 +/CD4 +and CD16 +CD56 +in peripheral blood were evaluated by FCM before and after treatment.Serum TNF- αand IL-10 levels were detected by the method of ELISA.Taking 3 sputum samples (night sputum,morning sputum and sputum at moment)of each patient to smear after treatment.Results Before treatment,there was no significant difference of each index between two groups.After treatment,CD4 +,CD4 +/CD8 +and CD16 +CD56 +in the observation group were significantly higher than that before treatment and the control group(P<0.05).The proportion of CD8 +in two groups was significantly lower than before treatment(P<0.05),the difference between groups was not statistically significant.Serum TNF- αand IL-10 levels decreased significantly in two groups than those before treatment and the control group(P<0.05).After treatment,the negative conversion rate in the control group was 90.9%(n=40),while 100%(n=45)in the observation group.The difference between two groups was statistically significant(P<0.05).Conclusion Mycobacterium vaccae vaccine combined with chemotherapy has a significant effect on children with pulmonary tuberculosis.It could significantly improve the immune function of children.

Objective To study clinical significance of anti-momoner C-reactive protein (anti- mCRP) antibody in systemic lupus erythematosus (SLE) and assess the relationship between serum CRP and anti-mCRP antibody.Methods Enzyme-linked immunosorbent assay (ELISA) was applied to determine serum level of anti-mCRP antibody in 113 pateints with SLE,65 patients with other rheumatic diseases,including primary Sjgren syndrome,rheumatoid arthritis,osteoarthritis,ankylosing spondylitis and systemic sclerosis,and 32 healthy controls.Serum level of CRP was evaluated by turbidimetry.Clinical manifestations and laboratory indicators of the patients were all recorded.Results Serum level of anti- mCRP antibody in SLE patients was significantly higher than that in patients with other rheumatic diseases and healthy controls,respectively (t=2.502 and 5.352,respectively,P 0.05).Titer of anti-mCRP antibody closely correlated with systemic lupus erythematosus disease activity index score (r=0.248,P0.05). Conclusions Level of Anti-mCRP antibody increased significantly in patients with SLE,which associated with disease activity of SLE and can be used as a valuable marker in evaluating activity of SLE.

BACKGROUND:In 1990s,overseas researchers use balloon occlusion method for establishing closed-chest animal models of myocardial infarction. But,ventricular fibrillation and thrombosis of intraoperative factors reduce the success rate of establishing the models. Currently,there are a few reports on establishing the large animal models. OBJECTIVE:We used balloon occlusion method for establishing closed-chest swine models of myocardial infarction,and explored ways to improve the success rate of modeling. DESIGN,TIME AND SETTING:The randomized controlled animal study of pathology observation was performed at the Department of Cardiology,First Affiliated Hospital of Kunming Medical College and Research Room of Pathology,Kunming Medical College from July 2008 to May 2009. MATERIALS:Fifteen Diannan small-ear pigs weighing 19-25 kg,aged 8-11 months,were divided into three groups:sham operation group,ischemia-reperfusion group,and ischemic postconditioning group,with 5 pigs in each group.METHODS:After the coronary occlusion and reperfusion period,the prophylactic use of lidocaine (1.0-2.0 mg/kg) infusion to control arrhythmia,and use of heparin to prevent and treat the thrombosis. A balloon catheter was positioned in the distal end of the first diagonal branch of the left anterior descending (LAD) artery under fluoroscopic guidance. In the sham operation group,the balloon was only placed to the LAD,did not block coronary artery. In the ischemia-reperfusion group,inflatable balloon occlusion was done for 60 minutes in the LAD after the balloon removed. In ischemic postconditioning group,after the balloon was inflated and occluded the LAD for 60 minutes,ischemic postconditioning was elicited by eight cycles of 30-second reperfusion,followed by 30-second reocclusion.MAIN OUTCOME MEASURES:Coronary angiography,electrocardiogram (ECG) and cardiac enzymes test was conducted to evaluate models of myocardial infarction. After three days,cardiac 2,3,5-triphenyltetrazolium chloride (TTC) staining and pathological examination was done to verily myocardial infarction.RESULTS:In the sham operation group,all pigs survived. In the ischemia-reperfusion group,4 pig models of myocardial infarction were successfully established,and one died of refractory ventricular fibrillation. In the ischemic postconditioning group,models of myocardial infarction after ischemia were successfully established. Following distal left anterior descending artery occlusion,the ECG leads V13 on the ST-segment elevation,the sick rational Q-wave formed;myocardial enzyme evolution of myocardial infarction in the human body was basically the same process. The site of myocardial infarction,basically the same parts,was located in apical,left ventricular anterior wall,and the former interval. TTC staining was normal myocardium brick red,myocardial infarct area appeared pale;pathological examination revealed a normal structure of myocardial infarct damage,cytoplasm condensed,dyeing deepening,transverse striations disappeared,nuclear enrichment,dissolution,fragmentation,many erythrocytes around the infarct area with abundant granulation tissue and a large infiltration of inflammatory cells.CONCLUSION:The described model presents a less invasion to the animals,and is the closest to the process of clinical practice.Intraoperative use of lidocaine and heparin for controlling arrhythmia and thrombosis of the models is successfully established as an effective manner. Ischemic postconditioning may be one of the factors for improving the modeling success rate.

Objective To explore the mechanism of deep brain stimulation(DBS)therapy to Parkinson's disease(PD).Methods We produced hemi-parkinsonian rat model with stereotaxically injecting 6-OHDA to right medial forebrain bundle(MFB)and stimulated ipsilateral subthalamu nucleus (STN)with platinum electrodes chronically to investigate the influence of DBS to the expression of Calbindin-28,synaptophysin and tyrosine dioxydase(TH)in Striatum by Western blot.In addition,slices of bilateral PD rats after DBS were stained to observe the expression of Calbindin-28 and synaptophysin in substantia nigra by Immunohistochemistry.Results High frequency stimulation impaired the rotational frequency 31% of unilateral PD rats triggered by apomophine;Long-term DBS increased the expression of TH in innocent striatum of unilateral PD rats 78.6%?9.5%,since the ipsilateral striatum(lesion side) was TH depleted by 6-OHDA insults;Calbindin-28 expression in ipsilateral striatum of hemi-PD rats raised up 75.4%?15.0% and long-term DBS reduced the effect by 43.0%?7.1%,meanwhile Calbindin-28 positive neurons in substantia nigra compacta in sham,PD and DBS rats were 74.5?10.2,75.7?15.6, 33.1?7.8.However,Synaptophysin expression in substantia nigra and striatum kept stable even after long- term DBS.Conclusions Consistent to the treatment to PD patients,DBS to STN alleviated the motor disorder of PD rats,the treatment might be based on regulating the expression of Calbindin-28 and TH.

Objective To study the clinicopathological and immunohistochemical features, histogenesis and biological behavior of solid pseudopapillary tumor of the pancreas ( SPT ). Methods Routine HE and immunohistochemical ( SP) methods were used in 20 cases of SPT. Results There were 18 females and 2 males, age ranging from 13 to 48 years with mean age of 25. 3 years. Abdominal pain and palpable mass were among the main complains. Seventeen cases were followed-up from 9 to 120 monthes. Fourteen cases were alive. Tumors were encapsulated, mixed with solid and cystic tissues. Histological features were psudopapillary structure with a fibrovascular core. Immunohistogically, the tumors were positive for a-1-AT ( 17 cases) , vimentin ( 14 cases) , synaptophysin ( 10 cases) , CgA (5 cases) ,CK and insulin (2 cases) ,glucagon and S-100 (1 case) ,PR (14 cases) , ER (1 case) ,pS2 (6 cases) , but all were negative for CEA and gastrin. Conclusion SPT is of low-graded malignancy and a distinct clinicopathologic entity in young female patients with both exocrine as well as endocrine differentiation. The tumor is closely related with sex hormone receptors.

BACKGROUNDNF-kappaB p65 was shown to inhibit transcription of phosphoenolpyruvate carboxykinase (PEPCK), a rate-limiting enzyme in gluconeogenesis in the liver. To understand the mechanism of action of NF-kappaB p65, we investigated the nuclear receptor corepressor in the regulation of PEPCK transcription.

METHODSRat H4IIE cells, human hepatoma HepG2 cells and human embryo kidney (HEK) 293 cells were used in this study. The transcriptional activity of a rat PEPCK gene promoter (-490/+100) was analyzed in HepG2 cells, a HepG2 super suppressor IkBalpha (ssIkBalpha) stable cell line, and HEK 293 cells. The effects of p65 and ssIkBalpha on a rat PEPCK gene promoter were observed using the PEPCK luciferase reporter system. The interaction of the cAMP-response- element-binding (CREB) protein, histone deacetylase 3 (HDAC3) and silencing mediator for retinoic and thyroid hormone receptors (SMRT) with the PEPCK gene promoter were investigated using the chromatin immunoprecipitation (ChIP) assay. p65 cotransfection and RNAi-mediated gene knockdown were used to determine the corepressor involved in the inhibition of PEPCK by NF-kappaB p65 and the transcriptional regulation of CREB by NF-kappaB p65.

RESULTSNF-kappaB p65 inhibited PEPCK expression and the inhibition was blocked by ssIkBalpha. The inhibitory effect of p65 was completely blocked in a HepG2 stable cell line in which ssIkBalpha was expressed. HDAC3 or SMRT knockdown led to a significant up-regulation of PEPCK reporter activity in the presence of p65 cotransfection. In the ChIP assay the interaction of HDAC3 and SMRT with the PEPCK gene promoter was induced by p65 activation, but the CREB signal was reduced. Transcriptional activity of CREB was inhibited by NF-kappaB p65 cotransfection. The inhibitory effect of NF-kappaB p65 was blocked by HDAC3 RNAi or SMRT RNAi.

CONCLUSIONSThe study showed that the inhibition of PEPCK by NF-kappaB p65 was dependent on HDAC3 and SMRT, which form a nuclear corepressor complex for transcriptional inhibition. The transcription factors NF-kappaB p65 and CREB share the same corepressor HDAC3-SMRT, and the corepressor exchange leads to inhibition of PEPCK gene transcription by NF-kappaB p65.

Animals ; Blotting, Western ; Cell Line ; Chromatin Immunoprecipitation ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Hep G2 Cells ; Histone Deacetylases ; genetics ; metabolism ; Humans ; NF-kappa B ; genetics ; metabolism ; Nuclear Receptor Co-Repressor 2 ; genetics ; metabolism ; Phosphoenolpyruvate Carboxykinase (ATP) ; genetics ; Promoter Regions, Genetic ; genetics ; Protein Binding ; genetics ; physiology ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factor RelA ; genetics ; metabolism