BACKGROUND:Bone marrow derived mesenchymal stem cells (BMSCs) will be homing to the lesions after balloon injury in the inflammatory reaction process.However,the molecular mechanism of transforming growth factor-β1 (TGF-β1) to promote BMSC into smooth muscle-like cell remains unclear.OBJECTIVE:To investigate the BMSC differentiation rate under different TGF-β1 levels after acute vascular injury.DESIGN,TIME AND SETTING:A randomized controlled animal experiment and in vitro induced cell observation were performed at the SPF Laboratory Animal Center and Laboratory of Tissue Engineering,Ninth People's Hospital of Shanghai Jiao Tong University Medical School between January 2008 and May 2009.MATERIALS:A total of 24 male SD rats were randomly divided into normal control and model groups with 12 animals in each group.In addition,6 4-weak-old male SD rats were used to prepare BMSCs.METHODS:Model of acute carotid artery injury was established in model group.Serum of normal control group and model group after 1,3,7 days of injury to detect TGF-β1 level after the vascular injury by ABC-ELISA.The BMSCs after one passage were cultured at a density of (1.0-3.0)×10~5 cells/100 mm in culture dish,and divided into two groups:in routine culture group,cells were cultured in high-glucose DMEM containing 20% fetal bovine serum;in TGF-β1 group,cells were induced by different concentrations of TGF-β1 (1,3,5 and 10 μg/L) based on routine culture for 1 week.MAIN OUTCOME MEASURES:Serum TGF-β1 level after the vascular injury;,cell morphological changes by inverted phase contrast microscopy;smooth muscle α-actin expression.RESULTS:The normal serum TGF-β1 level in rat was low,but increased rapidly after 1 day of injury,reached its peak at 3 days,and declined gradually but did not return to the basic level till day 7.After 1 week of induction,BMSCs were confluent,forming peak valley appearance.Immunocytochemistry showed that compared with routine culture,the rate differentiation of smooth muscle-like cell was significantly increased in cells stimulated by TGF-β1,especially 5 and 10 μg/L TGF-β1 (P ＜ 0.01).Real-time quantitative PCR results were similar to immunocytochemistry.CONCLUSION:Serum TGF-β1 level increased after acute vascular injury and peaked at day 3.In vitro,a similar concentration of TGF-β1 could induce cultured BMSC to differentiate into smooth muscle-like cells.

Objective To investigate the effective strategies to prevent and treat biliary complications after orthotopic liver transplantation.Methods The clinical data of 316 patients who received orthotopic liver transplantation at the Fuzhou General Hospital of Nanjing Military Command from November 2001 to March 2012 were retrospectively analyzed.Cold perfusion with HTK + UW solution was applied when obtaining the liver graft,and then the liver graft was preserved in the UW solution.The bile duct was perfused with UW solution thereafter.Orthotopic liver transplantation or piggyback liver transplantation were adopted in the cadaver liver transplantation.Left liver transplantation and right liver transplantation were adopted in the living donor liver transplantation.Choledochojejunal Roux-en-Y anastomosis or duct-to-duct choledochostomy were used for biliary reconstruction.Ordinary T tubes were used for drainage before 2006,and then 6 F pediatric suction catheter or epidural catheter were applied for drainage thereafter.The Ttube was pulled out 3-6 months after the operation.Enteral nutrition was applied to patients at the early phase after operation.The immunosuppressive agents used including tacrolimus + mycophenolatemofetil + adrenal cortical hormone,and for some patients,tacrolimus + mycophenolatemofetil + sirolimus + hormone were used.Patients were followed up for 2 years to learn the incidence of biliary complications and guide the medication.The difference in the incidence of bile leakage between patients who wcrc admitted before 2006 and those admitted after 2006 were compared using the chi-square test.Results The warm ischemia time was 2-6 minutes,and the cold ischemia time was 3-10 hours.For patients who received cadaver liver transplantation,orthotopic liver transplantation was carried out for 291 times and piggyback liver transplantation for 24 times; biliojejunal Roux-en-Y anastomosis was carried out for 5 times and bile duct end-to-end anastomosis for 310 times.For patients who received living donor liver transplantation,1 received left liver transplantation and 1 received right liver transplantation,and they received bile duct end-to-end anastomosis.A total of 311 patients received immunosuppressive treatment with tacrolimus + mycophenolatemofetil + adrenal cortical hormone,and 5 patients reveived tacrolimus + mycophenolatemofetil + sirolimus + hormone.Of the 316 patients who received orthotopic liver transplantation,38 had biliary complications after the operation,including bile leakage in 18 patients,intra-and extra-hepatic bile duct stricture in 6 patients,anastomotic stricture in 6 patients,biliarycomplications included cholangitis in the portal area and cholestasis in 4 patients,choledocholithiasis and cholangitis in 2 patients and biliary infection in 2 patients.The incidence of bile leakage before 2006 was 14.00％ (7/50),which was significantly higher than 4.12％ (11/267) of bile leakage after 2006 (x2-7.676,P ＜ 0.05).Of the 38 patients with biliary complications,the condition of 35 patients was improved,and 3 patients died.Of the 18 patients with bile leakage,15 was cured by conservative treatment,3 received surgical treatment (the condition of 1 patient was improved by drainage,anti-infection treatment and nutritional support,but died of peritoneal hemorrhage at postoperative 1 month; 2 patients received peritoneal drainage,1 was cured and 1 died of peritoneal infection).For the 6 patients with intra-and extra-hepatic bile duct stricture,1 was cured by liver retransplantation and 5 were cured by conservative treatment,endoscopic retrograde cholangio-pancreatography (ERCP) or balloon dilation.For the 6 patients with anastomotic stricture,the condition of 3 patients was improved by conservative treatment,balloon dilation or stent implantation,1 gave up treatment due to hepatic cancer recurrence and died thereafter,1 received anastomosis + T tube drainage,1 was cured by recurrent tumor resection and choledochojejunostomy.Four patients with cholangitis in the portal area and cholestasis were cured by conservative treatment.For the 2 patients with choledocholithiasis and cholangitis,1 was cured by stent implantation with ERCP,and 1 received conservative treatment,and the level of total bilirubin was decreased.Two patients with biliary infection were cured by anti-infection treatment.Conclusions Most of the biliary complications could be treated by non-surgical treatments.For patients with severe biliary complications or those could not be treated by non-surgical treatment,re-exploration of the bile duct is effective.Liver re-transplantation is the only choice for patients with dysfunction of liver graft caused by severe ischemic biliary injury.

Objective To evaluate the ultrasonographic ( US ) findings of granulomatous lobular mastitis( GLM ) ,and to compare the diagnostic accuracy among US ,mammographic ,and magnetic resonance imaging ( MRI) . Methods Imaging characteristics of 56 patients who were pathologically comfirmed as GLM were reviewed .All the lesions were assessed by BI‐RADS ( Breast Imaging Reporting and Data System) . Results Fifty‐eight lesions were found in 56 patients . Thirty ( 51 .7% ) were irregular ,20 (34 .5% ) were lobular and 8(13 .8% ) were round or oval in shape . Forty two lesions (72 .4% ) were hypoechoic ,14 (24 .1% ) were mixed echoic textur ,including 6 lesions (10 .3% ) with tubular connections and 8 lesions ( 13 .8% ) with irregular markly hypoechoic internal echoes . Two ( 3% ) were isoechoic .No calcification were found . Color Doppler signals were detected in 33 cases(56 .9% ) ,and the resistance index ( RI) ranged from 0 .61 to 0 .79 . Forty patients underwent mammography ,there were no distinct lesions in 6 cases(15% ) ,suspicious calcification in two(5% ) , asymmetric density in twenty(50% ) ,and solitary masses in twelve(30% ) . MRI was performed in 36 patients ,and revealed no abnormality in two patients(5 .6% ) , twenty nine lesions ( 80 .6% ) showed hypointensity on T1‐weighted images and hyperimensity on T2‐weighted images ,five lesions ( 13 .9% ) showed isointensity on T1‐weighted images and hyperimensity on T2‐weighted images ,and all the lesions showed heterogeneous enhancement after contrast .The diagnostic accuracy of ultrasonography , mammography and MRl was 63 .8% , 45% and 61 .1% , respectively . Conclusions There were no specific imaging characteristics of GLM ,the combination of ultrasonography , mammography and MRI might benefit the diagnosis of GLM .

Objective To analyze the ultrasonographic characteristics of breast cystic-solid lesions and to investigate the diagnostic value of BI-RADS breast imaging reporting and data system in this kind of disease Methods Imaging characteristics of 105 breast cystic-solid lesions of 93 patients were analyzed All the lesions were assessed by BI-RADS Histopathologic examination was used as the reference standard Results Among 105 lesions 57 54 3% were benign and 48 45 7% were malignant Differences in orientation shape margin calcification and vascularity were statistically significant between the benign and malignant lesions P < 0 05 whereas the echo pattern and the posterior acoustic feature did not show significant differences P >0 05).There were 5 1 predominantly solid mass,which appeared as solid mass with little dispersed or centralized cystic regions or solid mass with surrounding cystic regions,including 33 benign lesions and 1 8 malignant lesions.The area under the curves of ROC was 0.956 in this type.There were 25 lesions predominantly cystic,which had regular or irregular thick walls or septa,or had nodules protruding to the cystic component,including 1 1 benign lesions and 14 malignant lesions.The area under the curves of ROC was 0.753.There were 29 mixed solid-cystic mass,in which the solid component equaled to the cystic,including 13 benign lesions and 16 malignant lesions.The area under the curves of ROC was 0.887.The area under curves of ROC was 0.873 for the all breast cystic-solid lesions.The rates of malignant cystic-solid lesions in the three types had no significant difference(P >0.05).Conclusions Ultrasound is a useful tool for diagnosing breast cystic-solid lesions.The proportion of the cystic component is not helpful to differentiate malignancy from benignancy.

Objective To investigate the value of shear wave elastography (SWE)for differential diagnosis of benign and malignant thyroid nodules.Methods A total of 1 56 patients referred for surgery of thyroid nodules underwent standard ultrasound as well as elastography.The following characteristics of all the nodules were assessed:location,size,shape,margin,echogenicity,calcification and blood flow.The SWE elasticity indices of maximum (Emax),mean (Emean),minimum (Emin),and standard deviation (Esd) were automatically calculated by the US system.Thresholds were calculated by receiver operating characteristic (ROC ) curve analysis.The diagnostic performance of conventional US and SWE were compared.Histopathologic examination was used as the reference standard.Results Among a total of 1 56 nodules,1 1 8 were malignant and 38 were benign.SWE elasticity indices were significantly higher in malignant than in benign nodules (P ≤0.001 ).Sensitivity and specificity for predicting malignancy were 83.9%,92.1 % respectively using Emean with a threshold of 32.1 kPa,the specificity was significantly higher than using conventional ultrasound (P <0.05 ).Area under the ROC curve (Az)of Emean was highest (0.880,95% CI:0.81 7,0.943),however the differences were not significant(P >0.05).Conclusions SWE provides higher specificity for predicting malignancy without losing sensitivity.

The PIK3CA gene codes p100α,the catalytic subunit of phosphatidylinositol 3-kinase (PI3K) and is involved in the initiating the PI3K/AKT pathway.PIK3CA plays its biological roles through.downstream PI3K pathway. PIK3CA gene mutants can be detected in many kinds of tumors. The mutant PIK3CA gene can abnormally activate PI3K pathway,leading to the abnormal cell cycle,decreased cell adhesion,down regulated apoptosis and neovascularization,and then promotes tumor genesis and development.Recent researches have found that mutant PIK3CA gene is closely correlated with the genesis,development,differentiation,metastasis and drug resistance of colorectal cancer.Research of PIK3CA in colorectal cancer may provide significant evidence for the early diagnosis,gene screen,therapeutic regimen making,recurrence and follow up.

BACKGROUND:Caveolin-1 is expressed in mammalian brain and involved in the normal development of the brain, which can affect the proliferation of neural stem cells in the brain. OBJECTIVE:To acquire neural stem cells from caveolin-1 knockout embryonic mice in vitro and study their biological characteristics. METHODS:The whole brain was separated from C57BL/6 mice and caveolin-1 knockout C57BL/6 mice respectively at encyesis 14-16 days. Single cellsuspension was obtained by enzyme digestion, and cultured in the conditioned medium of neural stem cells. Fol owing 7 days of primary culture, the cells were induced in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum for 7 days. RESULTS AND CONCLUSION:The major cells of the cellsuspensions from the fetal mouse brain were dead at 1 day after culture, and some single cells floated in the medium and their transmittance were better, and then they gradual y formed multicellular bal s after 3 days. A smal amount of cells were adhered at the bottom of culture plate after passage, and a great amount of cellbal s appeared after 7 days. The proliferation rate of neural stem cells from caveolin-1 knockout mice was higher than that from normal mice. The cellbal s were nestin-positive and their differentiated cells was positive for neurofilament 200, glial fibril ary acidic protein or O4, respectively. Al of the cells from normal mouse brain were positive for caveolin-1, but the cells from caveolin-1 knockout mice were negative for caveolin-1 by immunocytochemistry. Moreover, the speed of cellbal formation and the number of cellbal s in neural stem cells from caveolin-1 knockout mice were better than those from normal mice. Caveolin-1 negative neural stem cells were cultured successful y from caveolin-1 knockout mouse brain, and the results show that caveolin-1 can promote the proliferation of neural stem cells and inhibit their differentiation in vitro.

Objective To evaluate the value of ultrasonic quantitative method in the diagnosis of liver fibrosis in chronic hepatitis B (CHB) patients. Methods Ultrasonography was performed in 186 CHB patients who underwent liver biopsies. Fifteen indices including liver capsule thickness and fourteen texture parameters of gray level co-occurrence matrix were extracted from standard sonograms and compared with fibrosis stages by histopathology. The status of liver fibrosis was divided into five stages from S0 to S4 by histopathology based on the disease severity. ANOVA and Spearman correlation analysis were applied to analyze the differences and relationships between these indices and pathological stage, respectively. Then discriminant analysis models were established based on the indices for quantitative diagnosis of liver fibrosis. Results Among the fifteen indices, including liver capsule thickness, only the variance (F=0. 55, r=0. 06; both P＞0. 05), sum average (F=0.61, both r=0.05 ; P＞0.05), sum entropy (F=1.68, r=0.09; both P≥0.05) and entropy (F=1.39,r=0.12; both P＞0.05) were not significantly associated with the stages and not manifested linear correlation. Using biopsy results as gold standard, the correct rank rate of discriminant analysis model analysis in the patients staged from S0 to S4 were 80. 0%, 64. 9%, 61.3%, 74. 1% and 80.6 %, respectively. There were 73.1% of cross-validated cases who were accurately classified by the model analysis. The sensitivity, specificity and accuracy in patients with stage ≥ 1 were 97. 6%,80.0% and 91.9%, respectively; those in patients with stage≥2 were 92.1%, 89.7% and 90.9%,respectively; those in patients with stage≥3 were 94.8%, 96.1% and 95.7%; and those in patients with stage 4 were 80. 6%, 97.4 % and 94.6%, respectively. When considered S0 as no fibrosis, S1 as mild fibrosis, S2 and S3 as moderate to severe fibrosis and S4 as early cirrhosis, the consistence rates between discriminant analysis model and biopsy result were 81.7%, 78. 4%, 56. 9% and 90.3%,respectively. There were 74.7% of cross-validated cases who were correctly classified. The sensitivity, specificity and accuracy of the models for determining the fibrosis severity in patients≥mild fibrosis were 97.6%, 81.7% and 92.5%, respectively; those in patients ≥ moderate to severe fibrosis were 83. 1%, 94.8% and 89.2%, respectively; those in patients with early cirrhosis were 90.3%, 93.5% and 93.0%, respectively. Conclusion As a novel and noninvasive method, ultrasonic texture analysis could quantitatively determine liver fibrosis in CHB patients and is worthy of further investigation.

Objective To explore effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide(VIP) in the hypothalamus, spinal cord and duodenum of chronical stressed mice. Methods Mice were randomly divided into normal, stress and Simotang group( n= 10 in each group), and given a variety of unpredictable chronic mild stress. After 21 days gastric emptying and intestinal propulsion function were measured,the expression of VIP was detected by immunohistochemistry and RT-PCR. Results Compared with mice in normal group( (49.81 ± 8.56)%; (51.02 ± 5.11 )% ), chronic stress increased gastric residual rate( (61.53 ±8.71 ) %; P ＜ 0.05 ) and reduced small intestine propulsion rate ( ( 31.79 ± 2.38 ) %; P ＜ 0. 05 ). There were differences in expressions of VIP positive cells and mRNA in duodenum( (8.8 ± 1.1 )/mm2 and(0. 58 ±0.03) ),hypothalamus ( ( 12.9 ± 1.5 )/mm2 and (0.81 ± 0. 07 ) ) and spinal cord ( ( 12.1 ± 1. 2)/mm2 and (0.76 ± 0.02) )in chronic stress group compared with normal group( (6.5 ± 0. 9)/mm2 and (0.43 ± 0. 04);( 10.8 ± 1.3 )/mm2and (0.57 ± 0.03 ); (9.3 ± 1.5 )/mm2 and (0.53 ± 0. 02 ) respectively). There was not difference in gastric residual rate (52.93 ± 9.15 )%, small intestine rate(48.98 ± 4.38 )% and expressions of VIP positive cells and mRNA in duodenum ( (6.7 ± 0.9)/mm2 and (0.48 ± 0. 05 ) ), hypothalamus ( ( 10. 6 ± 1.4 )/mm2 and ( 0. 61 ± 0. 05 ) )and spinal cord ( (9. 1 ± 1.3)/mm2 and(0.55 ± 0.05 ) ) in Simotang group compared with those in normal group (P ＞ 0.05 ), but there were decreased compared with those in chronic stress group (P ＜ 0.05 ). Conclusion Simotang can regulate expressions of VIP in duodenum, hypothalamusand spinal cord in chronically stressed mice.

Objective To discuss the effect of different methods of dropping the high temperature of the tubercular meningitis patients. Methods 35 cases with tubercular meningitis with high fever were divided into research group(11 cases) and control group(24 cases) ,24 patients in control group were treated by traditional cooling method, 11 patients in research group were rteated by mild hypothermia therapy, The self signs, the intracranial pressure,Glasgow scores, complications and the death rates of patients were assessed and compared. Results The effective rates of the research and control cooling method were 81.8% and 41.7% respectively. The difference between the two groups was significant;After the intervention,compared with the control group, the self scale is superior in research group,and the intracranial pressure and the death rates of patients were lower,and patients'Glasgow scores were higher,but the complications between the two groups had no significant differences. Conclusion Mild hypothermia can effectively control patients'temperature,at the same time, which can improve the patients conscious state and reduce the mortality.