Objective To investigate the possible influence of immunosuppressive therapy,including sirolimus (SRL) and calcineurin inhibitors (CNI, tacrolimus), on level of Treg in the liver allo-graft recipients, Methods Forty-seven liver transplant recipients with stable liver function were assessed for at least 2 years, and divided into two groups: one composed of 15 patients receiving SRL,and another, of those receiving CNI (32 patients with tacrolimus). Thirty-eight age-matched healthy subjects (HS) were used as normal controls. We examined the expression of CD4, CD25, and Foxp3 in peripheral blood monouclear cells. Treg of every group was analyzed using Cell Quest software.Results SRL significantly increased the prevalence of CD4+ CD25high T cells, compared with HS and CNI group. The prevalence of CD4+ CD25high T cells of CNIs group was significantly lower than that of HS. The percentage of CD4+ CD25high T cells in the total CD4+ T cells was 1.88 % (1.56 %-2. 60 %), 1. 15 % (0. 57 %-l. 48 %) and 0. 84 % (0. 46 %-1. 45 %) in SRL, NS and CNI group,respectively (P<0. 01 or P<0. 05). Foxp3 was expressed in more than 95 % of CD4 + CD25high T cells and less than 20 % of CD4+ CD25low T cells, whereas not expressed in CD4+ CD25- T cells.Conclusion Different immunosuppressive therapy (SRL or CNI) might have different roles in tolerance induction in liver transplant recipients. Namely, SRL promoted the induction of allo-antigen tolerance, but CNI hampered the progression.

Objective To investigate the application value of DFY-Ⅱ ultrasound imaging analysis software in evaluation of fetal lung maturity in different pregnancy.Methods Totally 315 cases of healthy single pregnancies (24-41 weeks) were collected.The sonographic views of fetal right side sagittal plane were obtained.The echo intensity of fetal lung and liver were analyzed and the ratio was canculated with DFY-Ⅱ ultrasound imaging analysis software.Results The ratio of echo intensity from fetal lung and liver had positive correlation with gestational weeks (r=0.94,P＜0.05),the linear regression equation was Y=0.60+0.07X (r2 =0.883).Conclusion The ratio of echo intensity by fetal lung and liver analyzed with DFY-Ⅱ ultrasound imaging analysis software can be an effective method to evaluate fetal lung maturity.

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.

Objective To investigate whether Tumor Necrosis Factor-alpha (TNFα) is capable of activating Rho kinase pathway which leads to smooth muscle cell proliferation and the intervention function of Rosuvastatin, and clarify the mechanism and intervention manner of anti-atherosclerosis by Rosuvastatin. Methods Wistar neonate rat smooth muscle cells were cultured, and the activity of cell proliferation was determined by methyl thiazolyl tetrazolium (MTT). The expression of Rho kinase genes after the stimulation of TNFα was evaluated by RT-PCR. Western blot method was used to measure the protein expression of proliferating cell nuclear antigen (PCNA) after TNFα stimulation and Rosuvastatin intervention in smooth muscle cell. Results The TNFα stimulation significantly enhanced the expression of Rho kinase and increased the expression of PCNA protein in smooth muscle cells (P < 0.05). These effects were positively correlated with prolonged treatment whereas additional Rosuvastatin administration inhibited the above-mentioned effects (P < 0.05). Conclusions The activation of TNFα mediated Rho kinase signaling pathway can significantly promote smooth muscle cell proliferation, and Rosuvastatin can not only inhibit this pathway but also the induced proliferation.

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.

In proteomic research, to improve protein solubility of membrane proteins and nuclear proteins, buffers containing high concentration of detergent, such as 4% SDS, were widely used. However, high concentration of detergent might severely interfere with the downstream proteomic analysis, including protein quantitation and trypsin digestion. To improve the proteomic compatibility of buffers with high concentration of detergent, we used short gel method to pretreat buffers containing detergent. Protein samples were first separated by a short (2-2.5 mm) SDS-PAGE electrophoresis, and proteins were quantitated by comparing with bovine serum albumin standards via optical density analysis. The gel was then cut and peptides were recovered using in-gel digestion. The quantitative linearity range of this method was 1 to 8 μg. The quantitation was accurate and reproducible. After short gel analysis, recovered peptides generated high mass spectrometry signals. In conclusion, short gel method eliminated the interference of high concentration detergent in the proteomics analysis, and it was suitable for protein samples' pretreatment, and was worth to apply in proteomic research.
Detergents ; chemistry ; Electrophoresis, Polyacrylamide Gel ; methods ; Mass Spectrometry ; Membrane Proteins ; chemistry ; Nuclear Proteins ; chemistry ; Proteins ; chemistry ; Proteomics ; methods ; Trypsin

Objective To explore the clinical characteristics and influencing factors of primary nephrotic syndrome (PNS) combined with hypercoagulability in children. Methods The clinical data of 57 children with primary PNS were analyzed retrospectively. The clinical features and treatment were compared among high coagulation state group, non high coagulation state group and control group (20 children). At the same time, the differences between the simple nephrotic syndrome group (SNS) and nephritic syndrome group (NNS) in hypercoagulable state were analyzed. In addition, the correlation analysis was performed. Results Among 57 patients, there were 50 patients in high coagulation state group and 7 in non high coagulation state group. There was no significant difference in gender, age and clinical manifestations between two groups (P>0.05). The platelet (PLT) count, platelet aggregation (PCT), albumin (Alb), fibrinogen (Fib), D-dimer (D2) were significantly higher than those in the control group, and there were statistically significant differences (P all<0.01). There were significant differences in the levels of PLT, Fib, D2 and complement C4 between hypercoagulable state group and non hypercoagulable state group (P all<0.05). There were significant differences in HCT, TC, LDL, PT and complement C3 levels between SNS group (n = 32) and NNS group (n =18) in 50 patients with high coagulation state (P<0.05). There was positive correlation between HCT and complement C3 (r=0.30, P<0.05), while there was no correlation between PLT and other indices (P>0.05). All of the 57 patients were improved and has no thrombosis after the treatment. Conclusion Children with primary PNS were usually associated with different degrees of hypercoagulable state, and PLT, Fib, D2 could be used as reference indices for the severity of hypercoagulable state, and the activation of complement system might be related to the occurrence and development of hypercoagulable state.

Objective To observe the perfusion pattern of hepatic angiomyolipomas using contrastenhanced ultrasound(CEUS) and compare diagnostic efficacy of contrast-enhanced ultrasound with contrastenhanced helical CT(CECT).Methods Nineteen patients with 21 resected and pathologically proven hepatic angiomyolipomas were included in this study.Low mechanical index (mechanical index less than 0.2) realtime CEUS was performed in nineteen patients (5 patients with pulse inversion harmonic, 14 patients with contrast pulse sequencing, CPS) after 2.4ml bolus injection of contrast agent SonoVue.CECT was performed using 16-slice helical CT and contrast agent Ultravist.The diagnostic performance was calculated by considering histologic results as the reference standards.Results Fifteen tumors were correctly diagnosed as hepatic angiomyolipomas,4 tumors were misdiagnosed (including 1 hepatocellular carcinoma,2hepatic adenomas, 1 hemangioma) and 2 tumors were characterized as benign lesions by CEUS.Eight tumors were correctly diagnosed as hepatic angiomyolipomas, 13 tumors were misdiagnosed (including 7hepatocellular carcinoma, 1 liposarcoma, 2 lipomas, 1 hepatic adenomas, 2 hemangiomas) by CECT.The preoperative diagnostic accuracy was 71.4% for CEUS and 38.1 % for CECT(P＜0.05 ).Conclusions CEUS can demonstrate typical imaging characteristics of most hepatic angiomyolipomas, and has higher diagnostic performance than CECT in characterization of hepatic angiomyotipomas.

Objective To investigate the risk factors associated with acute renal failure (ARF)after surgery in patients with simple abdominal trauma. Methods A retrospective case-control study was carried out in patients with simple abdominal trauma after surgery from January 2003 through May 2008 in our hospital. Twenty patients with ARF were set as study group and 68 patients without ARF as control group. Clinical data including age, gender, injury type, number of injured organs, blood transfusion volume, intra-abdominal pressure, abdominal infection, shock and related factors were analyzed by using univariate and Logistic regression to identify the independent risk factors of ARF after surgery in patients with simple abdominal trauma. Results Univariate analysis revealed statistical difference in aspects of blood transfusion volume ( ＞ 1 600 ml), operation timing ( interval between trauma and operation ＞12 h), preoperative shock duration ( ＞6 h), postoperative abdominal infection and intra-abdominal hypertension ( ＞ 12 mm Hg) between two groups ( P ＜ 0. 05 ). Logistic regression analysis showed that these five indices were the independent risk factors of ARF after surgery in patients with simple abdominal trauma ( P ＜ 0. 05 ). Conclusions Massive blood transfusion, delayed operation, long preoperative duration of shock, postoperative abdominal infection or intra-abdominal hypertension are the risk factors of ARF after surgery in patients with simple abdominal trauma. We should pay attention to these factors and take effective measures to prevent occurrence of ARF.