Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.

Objective To investigate the blood-saving effect of controlled low central venous pressure (CLCVP) in different types of hepatectomy.Methods Ninety ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 37-76 yr,weighing 40-75 kg,undergoing elective hepatectomy,were divided into 6 groups according to the surgical approach and whether CLCVP was used during surgery (n =15 each):CLCVP1-3 groups and nonCLCVP1-3 groups (NCLCVP1-3 groups).The standard hepatectomy,half liver resection and irregular hepatectomy were performed in CLCVP1-3 groups,respectively,with CLCVP.The standard hepatectomy,half liver resection and irregular hepatectomy were performed in NCLCVP1-3 groups,respectively,without CLCVP.In CLCVP1-3 groups,from skin incision to the end of liver resection,CVP was maintained ≤ 5 cm H2 O through adjustment of the position,fluid restriction and iv infusion of nitroglycerin,and norepinephrine was infused simultaneously to maintain mean arterial pressure ≥ 60 mm Hg.In NCLCVP1-3 groups CVP was maintained at 6-12 cm H2O.Intraoperative blood loss and blood transfusion were recorded.Results Compared with NCLCVP1-3 groups,intraoperative blood loss was significantly decreased in CLCVP1-3 groups (P ＜ 0.05).Compared with NCLCVP3 group,the amount of blood transfusion was significantly decreased,the constituent ratio of intraoperative blood loss ＜ 200 ml was increased,and the constituent ratio of intraoperative blood loss ＞ 1000 ml was decreased in group CLCVP3 (P ＜ 0.05).Conclusion CLCVP can decrease the intraoperative blood loss and blood transfusion in patients undergoing irregular hepatectomy.

Objective To study the effect of SYUIQ-5,a novel antitumor agent,on gene expression of hepatic cytochrome P450 in rats.Methods Male Sprague-Dawley rats were administered daily intraperitoneal dose of SYUIQ-5(0.1mg/kg,5mg/kg and 10mg/kg body weight) for 5 consecutive days.The levels of CYP 3A1,2B1/2,1A1,1A2 and 2E1 mRNA in rat liver were analyzed by reverse transcription-polymerase chain reaction(RT-PCR).Results The level of gene expression of CYP 1A1 was increased significantly compared with the control.The levels of CYP 3A1,2B1/2,1A2 and 2E1 mRNA were not changed by SYUIQ-5 treatment compared with the control.Conclusion A significant increase in the CYP 1A1 rather than CYP3A1,2B1/2,1A2 and 2E1 gene expression by SYUIQ-5 treatment is observed.

0.05). Cross-reactivity was seen in 1 case of clonorchiasis sinensis by both the methods. Conclusion Microwave ELISA has the advantages of high sensitivity, specificity and rapidity.

There are many subjects related to anesthesiology and the band between anesthesiology and clinical practicevery is close. So students can not be proficient in anesthesiology by traditional teaching methods. The use of three-dimensional case teaching method in anaesthesiology teaching is a good attemptment. three-dimensional case teaching method integrate case method, multimedia technology and clinical case. It can help medical students analyze and solve clinical problems, improvetheir academic performance and enhance their clinical basic skills.

Objective: The bacterial superantigen staphylococcal enterotoxin A (SEA) has the potent ability of inducing T cell activation as well as directing activated T cells to kill tumour cells. However, its application in the tumour treatment is limited due to its systemic immune activation.In order to minimize its side effects SEA liposomes is prepared and their toxcicity in vivo is investigated . Methods: SEA liposomes were prepared by the method of reverse-phase evaporation . SEA liposomes were administered intravenously . In vivo the toxcicity of SEA liposomes was investigated by the measurement of blood pressure, colonic temperature and breath rate of New Zealand rabbits. Mice plasma TNF-? and IFN-? level were determined by ELISA . Results:After iv administration of SEA liposomes, significant reduction of mice plasma TNF-? and IFN-? level was observerd. As compared to free SEA, liposomal SEA had less effect on blood pressure, colonic temperature and breath rate of the rabbits. Conclusion:SEA Liposomes had relatively low toxicity. These advantage was probably due to its lower systemic immune activation effect and inducing consequent lower systemic TNF-? and IFN-? level.

ObjeCtive To investigate the value of protein biochip in the diagnosis of Lyme borreliosis. Methods The serum IgM and IgG antibodies against flagellin were detected by flagellin coated-and N-hydroxysuccinimide (NHS) modified-biochips in 82 patients with neuroborreliosis (NB),35 patients with erythema migrans (EM) and 44 normal controls.The results were compared with those from enzyme-linked immunosorbent assays (ELISA) and Western blot.Results The levels of both IgM and IgG antibodies were significantly higher in the NB patients than those in the normal controls (P

OBJECTIVETo investigate the effect of four traditional Chinese medicines distributed along lung meridian, namely Ephedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex, on TLR2 and NF-κB expressions in mice with lung heat syndrome, in order to study the intervention effect of the four traditional Chinese medicines (TCMs) on the lung heat syndrome.

METHODOne hundred KM mice were randomly divided into the normal control group, the model group, the Ephedrae Hebra group, the Zingiberis Rhizoma group, the Scutellariae Radix group and the Mori Cortex group (20, 10 g x kg(-1)), nasally dripped with streptococcus pneumoniae to establish the mouse lung heat syndrome model, and then administered with different TCMs. The expressions of TLR2, NF-κB p65 proteins in lung tissues were analyzed by the immunohistochemical method. The expressions of TLR2, NF-κB p65 mRNA were measured by real time PCR.

RESULTCompared with the normal control group, the expressions of TLR2 and NF-κB p65 proteins in lung tissues in the model group were higher (P < 0.01), and the expressions of TLR2 and NF-κB p65 mRNA in lung tissues were up-regulated (P < 0.01 or P < 0.05). Compared with the model group, Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group and the Scutellariae Radix high dose group showed decreased expression of TLR2 protein (P < 0.05 or P < 0.01); Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group, Scutellariae Radix high and low dose groups and Mori Cortex high and low dose groups showed reduced expression of NF-κB p65 protein (P < 0.05 or P < 0.01). Ephedrae Hebra high and low dose groups, Zingiberis Rhizoma high and low dose groups, Scutellariae Radix high dose group and Mori Cortex high dose group showed down-regulated expression of TLR2 mRNA (P < 0.01 or P < 0.05).

CONCLUSIONEphedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex can induce the TLR2/NF-κB inflammatory signal pathways by down-regulating the expressions of TLR2 and NF-κB p65 in protein and mRNA, so as to alleviate the lung tissue injury in mice with lung heat syndrome.

Animals ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; classification ; pharmacology ; Female ; Gene Expression ; drug effects ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; microbiology ; Male ; Medicine, Chinese Traditional ; methods ; Meridians ; Mice ; Phytotherapy ; methods ; Plants, Medicinal ; chemistry ; classification ; Pneumonia, Pneumococcal ; drug therapy ; genetics ; metabolism ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria baicalensis ; chemistry ; Species Specificity ; Syndrome ; Toll-Like Receptor 2 ; genetics ; metabolism ; Transcription Factor RelA ; genetics ; metabolism

Objective To investigate the effect of controlled low central venous pressure(CLCVP) on blood loss and prognosis in different types of hepatectomy .Methods Two hundred and fifty seven patients underwent standard hepatectomy ,half liver resec‐tion or irregular partial hepatectomy from January 2011 to December 2012 in the First Affiliated Hospital of Chongqing Medical U‐niversity were retrospectively studied .Patients treated with CLCVP during hepatectomy were attributed to the CLCVP group .CVP of these patients were lowed to below 5 cm H2 O by minimizing fluid infusion and one or both of the following maneuvers :posture adjustment ,nitroglycerin administration .Alpha agonists were used when necessary to maintain the mean arterial pressure MAP at ≥60 mm Hg .Other patients been maintained with normal level of CVP by adjusting fluid administration were included in normal CVP group (NCVP) .Blood loss and transfusion volume ,length of hospital stay of the two groups were compared ,and the effects of different surgery type on CLCVP blood protection were evaluated .Results In the patients underwent standard hepatectomy or half liver resection ,intraoperative blood loss and transfusion were not statistically different between the two groups .While in the pa‐tients underwent irregular partial hepatectomy ,the CLCVP group suffered less blood loss and transfusion(P<0 .05) .Percentage of the patients with less than 200 mL blood loss and no transfusion of concentrated red cell in CLCVP group was higher than that of in NCVP group(P<0 .05) .Differences between the two groups in postoperative hospital stay were with no significance in all the operation types(P>0 .05) .Conclusion The efficiency of CLCVP on blood protection during hepatectomy is influenced by the sur‐gery type ,the blood protection is found to be significant only in irregular partial hepatectomy .No relationship was found between CLCVP and postoperative hospital stay in all types of hepatectomy .

Objective To investigate the expression of endothelial protein C receptor (EPCR) and its roles in plasma and placenta of patients with early onset severe preeclampsia. Methods Sixty cases of severe preeclampsia women who delivered in Xuzhou Maternity and Child Health Care Hospital from March 2014 to February 2016, were recruited, which included 30 cases with early onset severe preeclampsia (early onset group, gestational week <34 weeks ) and 30 patients with late onset severe preeclampsia (late onset group, gestational week ≥34 weeks). Thirty cases of healthy late pregnant women at the same period (gestational week≥34 weeks) were selected as control group. Immunohistochemistry SP method was applied to detect the expression of in EPCR placenta. Reverse transcription (RT)-PCR was used to detect the expression of EPCR mRNA in placenta. ELISA method was used to detect the levels of soluble EPCR (sEPCR)level in plasma of the pregnant women of the three groups. Results The expression of EPCR in placenta mainly distributed in the membrane and cytoplasm of placental syncytiotrophoblasts and vascular endothelial cells, a few in the cell nucleus. The expression of EPCR in early onset group(57%, 17/30)was significantly lower than that in late onset group (93%, 28/30; χ2=25.165,P=0.001). The expression of EPCR in late onset group had no significant difference from that in control group (97%, 29/30;χ2=0.540,P=0.910). The expression of EPCR mRNA in placenta of early onset group(0.40±0.07)was significantly lower than that in late onset group(0.91±0.06;t=-30.044,P=0.001), while there was no statistical difference of the expression of EPCR mRNA between the late onset group and the control group (0.92±0.07;t=-0.631, P=0.538). Plasma sEPCR level in early onset group, late onset group and control group were (231 ± 11), (124±6)and(121±4)μg/L respectively, which is higher in early onset group than that in late onset group (t=48.080,P=0.001). There was no statistical difference of plasma sEPCR level between the late onset group and the control group(t=2.534,P=0.100). Conclusions The pathogenesis of early onset and late onset severe preeclampsia may be different. Decreased expression of EPCR in placenta may be associated with the pathogenesis of early onset severe preeclampsia.