Objective To explore the application value of 3.0T magnetic resonance perfusion weighted imaging(PWI)in the rat late brain glioma model.Methods C6 glioma cells were injected stereotactically into the right caudate nucleus of 32 Wistar rats.In 3 weeks after inoculation,the conventional MRI and PWI were performed.The relative cerebral blood volume(rCBV)values were recorded and compared with the pathological results.Results The tumor formation in 30 inoculated rats were confirmed by the sub-sequent pathological examination,with the tumor formation rate of 100%.PWI showed that rCBV values of the brain glioma region and the contralateral mirror image brain tissue were 262.61±72.82 and 189.39±57.21,the difference between them was statisti-cally significant(P<0.05).Conclusion The rat late brain glioma model is suitable for the study on the blood volume of brain tumor tissues.

Objective To analyze the skin barrier function in patients with facial steroid dermatitis.Methods Ninety-five patients with facial steroid dermatitis were enrolled into this study along with 25 healthy volunteers as controls.Transepidermal water loss(TEWL)and skin hydration were measured in 8 facial regions.including lesional skin in forehead,nasal tip,submaxilla,both cheeks and angulus oris,and non-lesional skin in the left posteior ear.Results Compared with the healthy group,the patients with steroid dermatitis had a significant higher value of TEWL on lower mandible,both angulus oris and cheeks(t:4.90,2.60,2.57,2.54,3.77,respectively,P＜0.01),while a significant lower level of skin hydration was noted on both angulus otis,right cheek and forehead(t=3.27,3.81,2.02,2.78,respectively,P＜0.05).Among the 8 test sites in both patients and controls,TEWL value decreased in the following order:submaxilla and both angulus oris＞both cheeks and forehead＞non lesional skin on the posteroir ear,and the decrease in skin water content was highest in submaxilla and both angnlus oris,followed by cheeks and forehead,and finally by left posterior ear.Conclusions In patients with facial steroid dermatitis,skin barrier function is disturbed on submaxilla,both angnlus oris and cheeks,and TEWL value is found to be higher in perioral area,while cheeks have a lower level of skin hydration.

A new method based on adherence of cellulolytic bacteria to insoluble cellulose for isolation and purification of thermophilic cellulolytic anaerobes was reported, in which Hungate anaerobic operating techniques were used to roll tubes with insoluble cellulose powder as substrate.

Objective:To clone soluble human anti-digoxin antibodies from large phage antibody library and construct a vector that expresses diabody.Methods:Soluble ScFvs were prepared through infecting E coli.HB2151 with the selected phage antibodies and induced with IPTG.The diabody vector was modified by enzyme digestion and checked by SDS-electrophoretogram.Results:It was showed by ELISA that soluble ScFv had specific binding ability to digoxin.The vector to express a soluble diabody was obtained by genetic modification,which was shown by Western blot.Conclusion:Soluble human anti-digoxin antibodies were successfully obtained from phage antibodies.The vector is efficient in creating diabody.

Periampullary carcinoma is a rare malignant tumor of digestive system,and its accurate diagnosis is still difficult.From January 2007 to July 2012,12 patients with periampullary carcinoma had been admitted to the Southwest Hospital of Third Military Medical University,and the imaging data were retrospectively analyzed.The results of ultrasonography revealed that all tumors were hypoechoic.The tumor displayed hyperenhancement in 3 patients,isoenhancement in 1 patient,hypoenhancement in 8 patients during the arterial phase on contrastenhanced ultrasonography (CEUS),while the tumor displayed hypoenhancement in all patients during the venous phase.Magnetic resonance imaging (MRI) plain scanning showed duodenal papilla enlargement in 1 patient,ampullary tissue mass signal in 2 patients,tissue mass signal at distal common bile duct in 2 patients,the rest 7 patients did not show tissue mass signal.Lower biliary obstruction was the common manifestation of the 12 patients on magnetic resonance cholangiopancreatography (MRCP),intrahepatic and extrahepatic bile vine-like expansion in 4 patients,double duct sign in 2 patients,the bottom of common bile duct with filling defect in 2 patients and it revealed beak-like narrow in 1 patient.CEUS,MRI and MRCP could both play an important role as conventional methods in diagnosing periampullary carcinoma.

Objective To isolate,purify and identify pancreatic duct derived stem cells (PDSCs) from the pancreatic duct of rats,and culture in the three-dimension cell culture system.Methods Adult male Wistar rats underwent perfusion with collagenase V via the pancreatic duct,then the pancreas was surgically procured,digested,followed by discontinued density gradient centrifuge to isolate ductal tissue from islets.The acinar and ductal tissue was cultivated in serumcontaining medium in the three-dimension cell culture to obtain adherent cells,as PDSCs,which were expanded by consecutive passages.The morphology and characterization of PDSCs on phenotype were examined.Results PDSCs could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three dimension cell culture system.Morphologically,PDSCs had remarkable size,most with one nucleus.PDSCs grew in many layers in three-dimension cell culture system.PDSCs was revealed to express CD29,CD73,CD90,CD105,but not CD14,CD19,CD34,CD45 by FACS,in agreement with MSCs.Conclusion PDSCs of rats could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three-dimension cell culture system.PDSCs lines were successfully established.

AIM:To discuss the effect of Shenmai injection on insulin resistance ( IR) in 3T3-L1 cells and its mechanisms.METHODS:3T3-L1 preadipocytes were induced by chemical reagents to differentiate into fully differentiated adipocytes.Oil red O staining was used to detect the differentiation level of the adipocytes .The insulin-resistant 3T3-L1 cell model was demonstrated using insulin , which was confirmed by glucose concentration in cell supernatant .The IR cell model was given 10 μmol/L rosiglitazone , 25 and 50 g/L Shenmai injection and normal saline for comparison .MTT assay was used to assess the cell activity of 3T3-L1 cells which was treated with drugs for 8, 16, 24 and 36 h.Glucose oxidase method was used to detect the glucose concentration in the cell supernatant at 8, 16 and 24 h.The protein levels of glucose transporter-4 (GLUT4), phosphatidylinositol 3-kinase (PI3K), AKT and p-AKT were determined by Western blot .RE-SULTS:3T3-L1 adipocytes were successfully induced as shown by the positive oil red O staining .The IR cell model was demonstrated , and glucose concentration in the cell supernatant after treatment with Shenmai injection showed that Shenmai injection reduced the IR in 3T3-L1 cell model.The protein levels of GLUT4, PI3K and p-AKT increased compared to con-trol group.CONCLUSION:Shenmai injection reduces the IR in 3T3-L1 cell model, which functions by increasing the protein levels of GLUT4, PI3K and p-AKT.

Objective To determine the expression of indole-2,3 dioxygenase (IDO) in human acute leukemia,and to investigate its correlations with clinicopathological parameters and prognosis in acute leukemia.Methods The expression of IDO in protein and RNA levels was detected by immunohistochemistry and real-time quantitative RT-PCR,respectively,and the correlations of IDO with clinicopathologic features and prognosis of acute myeloid leukemia (AML)-M5 were analyzed.Results The positive rate of IDO protein was 63.3 ％ (38/60) in human acute leukemia,while it in AML (34/49,69.4 ％),especially in AML-M5 patients (29/35,82.9 ％),was significantly higher than that of acute lymphoblastic leukemia (4/11,36.4 ％).The expression of IDO protein in healthy human peripheral blood mononuclear cells was negative.The RNA expression level of IDO in AML-M5 or non AML-M5 patients were significantly higher than that of healthy people (P ＜ 0.001),and AML-M5 patients had significantly higher IDO RNA level than that in non AML-M5 patients (P ＜ 0.05).The IDO gene expression was not correlated with sex,age and drug sensitivity,while it was closely related with these factors in the patients without complication of pulmonary infection.IDO could not act as an independent prognostic marker.Conclusion The expression of IDO in AML-M5 patients is significantly higher than that in non AML-M5 patients and healthy people.The positive expression of IDO is associated with poor prognosis of AML-M5 patients,but it is not an independent poor prognostic indicator.

Objective To explore the correlation of the myeloid antigen expression and clinical characteristics of acute lymphoblastic leukemia (ALL) in children.Methods The clinical data of 77 newly diagnosed ALL patients in Department of Hematology,the People's Hospital of Zhengzhou University from Jan.2010 to Dec.2013 were analyzed.The patients included 53 boys and 24 girls with a median age of 7.73 (2.00-15.00) years old.Based on flow cytometry (FCM) analysis of bone marrow,these patients were divided into 2 groups:one group included 26 patients with positive myeloid antigen expression (MyAg + ALL) and the other group included 51 patients with negative myeloid antigen expressions (MyAg-ALL).The correlation among myeloid antigen expression,clinical features,prednisone experiment,myelogram on the 15th day was analyzed through induction chemotherapy and minimal residual disease (MRD) on the 33rd day,and the rate of disease-free survival (DFS) was compared between the 2 groups.Results There were 26 cases with myeloid antigen expression among 77 patients (33.77％),CD13 + accounting for 19.48％ (15/77 cases),CD33 + 10.39％ (8/77 cases),and CD117 + 5.19％ (4/77 cases).Among these patients,there were 2 patients expressing both CD13 + and CD33 +,and 1 patient expressing both CD33 + and CD117 +.There was no difference between the MyAg + ALL group and MyAg-ALL group in gender (x2 =0.217,P =0.641),age (≥ 10 years old,x2 =0.011,P =0.918),white blood count(≥50 × 109/L,x2 =1.198,P =0.274),lactate dehydrogenase (LDH) (≥500 U/L,x2 =0.317,P =0.573),genetic abnormality (x2 =0.377,P =0.539),immunophenotype (B-ALL/T-ALL,x2 =0.397,P =0.529),and risk stratification (low-risk group,middle-risk group and high-risk group,x2 =0.260,P =0.878).Univariate Logistic regression showed that the reaction rate of prednisone experiment (P =0.023,OR =3.422) and positive rate of MRD (P =0.001,OR =0.133) of MyAg + ALL group were obviously higher than those in MyAg-ALL group.Multivariate Logistic regression showed that positive rate of MRD in CD13 + ALL group was obviously higher than that of CD13-ALL group (P =0.034,OR =120.765).The DFS rate of CD13 + ALL group and CD13-ALL group were (50.4 ± 13.8)％ and (77.4 ±6.7)％ respectively,and there was a significant difference between the 2 groups (x2 =3.928,P =0.047).Conclusions There is no significant correlation between myeloid antigen expression and clinical characteristics of children patients with ALL.For the patients with myeloid antigens,the early reaction of induction chemotherapy is bad,and for patients with CD13,the prognosis is not good.