Objective To study the binding performance of 99Tcm labeled anti-human prostatic specific membrane antigen (PSMA) monoclonal antibody J591 (99Tcm-J591) and prostate cancer cells in vitro,the biodistribution and SPECT imaging of 99Tcm-J591 in nude mice bearing human prostate cancer in vivo.Methods The monoclonal antibody J591 was labeled with 99Tcm by improved Schwarz method.Labeled antibody was purified by Sephadex G-50.The labeling efficiency and radiochemical purity were measured by paper chromatography and trichloroacetic acid method.The binding performance of J591 and prostate cancer cells was measured by flow cytometry in vitro.The nude mice bearing PSMA-positive C4-2 prostate carcinoma xenografts served as experiment group,mice bearing PSMA-negative PC3 tumors served as control group.6.2-8.5 MBq of 99Tcm-J591 (25 μg) was intravenously injected into mice.Gamma imaging was performed 2,6,12 and 24 h after injection,T/NT was calculated by ROI technique.After scanned 12 h post injection,4 mice of the experiment group and 5 mice of the control group were sacrificed and the tracer in vivo biodistribution was measured by gamma-counting,and the ％ ID/g was calculated.Two-sample t test was carried out to validate significant difference of ％ID/g between two groups.Results The labeling efficiency and radiochemieal purity of 99Tcm-J591 were (78.9±6.2)％ and (92.3±5.1)％,respectively,and the specific activity of 99Tcm-J591 was 68.7 MBq/mg.The antibody J591 and 99Tcm-J591 could strongly combine with PSMA-positive C4-2 cells in vitro,and didn't combine with PSMA-negative PC3 cells in vitro.SPECT imaging results showed that radioactive concentration was obvious in tumor 6 h post injection,the concentration scope became large and the tumor image was clear 12 h post injection.T/NT was 1.9±1.1 at 2 h,4.3±1.8 at 6 h,5.6±2.7 at 12 h,1.4±0.6 at 24 h,respectively.In the control group,no radioactivity concentration was found in tumor,and T/NT was less than 2.The biodistribution results showed that ％ID/g of tumor tissue was 20.1±5.2 in the experiment group and 5.8±2.6 in the control group,and there was significant difference (t=5.37,P＜0.001).No significant tracer uptake occurred in other tissues and organs between the two groups (all t＜ 1.98,all P＞0.05).Conclusion The immunoactivity,characteristics of biodistribution and tumor targeting property of monoclonal antibody J591 show promising future and potential values in diagnosis and therapy of prostate cancer.

Objective To prepare Exosomes secreted by mouse hepatoma cell (H_(22)) and heat stressed Exosomes (HS-Exo) derived from heat stress-treated mouse hepatoma cell (H_(22)), in order to study the possible anti-tumor immune mechanism. Methods Exosomes and HS-Exo were purified by serial ultracentrifugation and sucrose density gradiant centrifugation, and were observed and identified by electron microscope. The components and production of the protein and the effects of the host immune response against hepatocellular carcinoma of HS-Exo were observed by using Exosomes as the control. Their immunological factors were detected by Western blot. Lymphocyte proliferation and specific cytotoxic activity of mouse splenic cells were determined by MTT. CD4~+ and CD8~+ lymphocytes infiltration in mouse tumor tissues immunized by both were analysed by immunohistochemical staining. Results HS-Exo was similar in morphology to the Exosomes, the important immune-ralated protein expressed in HS-Exo was increased (P<0.05). HS-Exo immunized mouse group showed more effective inhibition of tumor growth, better-induced lymphocyte proliferation,more significantly enhanced the cytotoxic activity of spleen lymphocytes, as well as a more prominent role in tumor therapy than Exosomes immunized mouse control group (P<0.05). Conclusions Heat stress treatment method for the preparation of HS-Exo was feasible. HS-Exo had a stronger role in the immuneactivity and tumor treatment than control Exosomes.

Objective To construct a lentiviral vector carrying CRKL gene RNA interfering( RNAi ).Methods The CRKL RNAi was selected and subcloned into the lentiviral vector,pGCL-GFP(including U6 promotor and green fluorescent protein),generating the lentiviral vector LV-shCRKL.The corrected CRKL was confirmed by endoenzyme digestion ,sequencing.Recombinant lentiviruses were produced by 293T cells following the eo-transfection of LV-shCRKL,with the packaging plasmids pHelper1.0 and pHelper2.0.The virus titer was detected by GFP expressions in 293T cells.Results Plasmid LV-shCRKL carried the correct sequence.The recombinant lentiviruse LV-shCRKL could be produced by co-transfection of LV-shCRKL to 293T cells.Conclusion The recombinant lentiviruse vector LV-shCRKL is constructed successful.

Objective To explore the mechanism and way for CT10 regulator of kinase like (CRKL) involving in drug resistance in leukemia cells. Methods The four major proteins included Ras protein, signal transducer and activator of transcripton 5 (STAT5) protein, phosphoinositide 3-kinase (PI3K) protein and paxillin protein in leukemia which involved in signal transduction pathway of CRKL. The expressions of those proteins were detected by Western-blot and immunofluorescent staining and confocal laser scanning microscopy. Results Compared with K562/S cells, the expressions of Ras(41.52±15.47 vs. 23.74±8.67) and PI3K (35.60±12.48 vs. 10.09±0.005) protein were up-regulated in K562/ADM cells (t=3.01,6.13;both P<0.05), while there were no significant changes in the expressions of paxillin (20.10±11.89 vs. 23.11±12.40) and STAT5 protein (25.72±14.46 vs. 17.58±9.21) between K562/S cells and K562/ADM cells(t=0. 18,1.43;both P>0. 0S). Conclusions Ras and PI3K protein may play a role in the multidrug resistance of K562 cell line, while paxillin and STAT5 protein may be not involved in the formation of resistant in K562 cells.

According to the Organ Injury Scale Grading System of the American Association for the Surgery of Trauma (AAST-OIS),grade Ⅴ liver trauma is always complicated with retrohepatic inferior vena cava injury and less bile duct injury,and it is extremely severe and difficult to be treated.Timely and fast judgment,emergent exploration and effective repair of the injured bile duct are the key points for the treatment of bile duct injury.One patient with grade Ⅴ liver trauma combined with hilar bile duct transection injury was admitted to the Fuzhou General Hospital of Nanjing Military Command on August 30,2013.The rupture of left and right liver junction was detected by preoperative multidisciplinary consultation and emergency open surgery at admission hour 4.There was left and right hepatic duct bifurcation rupture at the first hepatic hilum.Non-functional liver tissues were excised.Breakage left and middle hepatic vein were sutured by polymer suture line.Liver traumatic bleeding and bile duct were sutured and ligatured individually.Left and right hepatic duct laceration was sutured by 6-0 PDS suture line.A hole in the stomach wall was opened fist,and then most part of the gastric contents was removed and the gastric wall was reparied by stapler.Patient received the postoperative symptomatic treatment with gradual recovery,and was discharged from hospital at admission day 26.The patient was readmitted to the hospital at 31 days of discharge due to outflow of purulent fluid from abdominal cavity drainage tube,and was treated by ceftriaxone sodium and tazobactam sodium according to the results of drug sensitive test and continuous peritoneal lavage.The abdominal cavity drainage tube and left and right hepatic duct drainage tube were removed at postoperative day 83.The patient was discharged from hospital at readmission day 28,and was followed up till December 2014 with good recovery and without complication.

Objective To compare the blood-saving effect when acute hypervolemic hemodilution (AHH) was performed with hydroxyethyl starch (HES) 130/0.4 dissolved in electrolyte injection (HES-E) and HES 130/0.4 in sodium chloride injection (HES-NaCl).Methods Thirty patients of both sexes,aged 18-60 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,with body mass index of 18-25 kg/m2,hemoglobin (Hb) ＞100 g/L,hematocrit (Hct) ＞ 35％,scheduled for elective abdominal operations under general anesthesia,were randomly divided into HES-E group and HES-NaCl group using a random number table,with 15 patients in each group.AHH was performed after induction of anesthesia.In HES-E and HES-NaCl groups,HES-E and HES-NaCl 15 ml/kg were intravenously infused over 30 min,respectively,and the infusion was conpleted before skin incision.Immediately after onset of AHH (T1),at 2 h after the end of AHH (T2),and at the end of operation (T3),arterial blood samples were collected for blood gas analysis and blood routine test,and pH value,base excess,HCO3-,K+,Na+,Cl-,Ca2+,Hb and Hct were recorded.Venous blood samples were collected at T1 and T2 for measurement of blood coagulation parameters including prothrombin time,activated partial thromboplastin time and fibrinogen and thrombelastography parameters.The volume of liquid intake and output and requirement for allogeneic blood transfusion were recorded,and the blood volume expansion rate was calculated.Results Compared with group HES-NaCl,no significant changes were found in the total volume of liquid infused,requirement for allogeneic blood transfusion,blood volume expansion rate,blood coagulation parameters at each time point,Hb and Hct (P＞0.05),pH value,base excess,HCO3 and K+ were significantly increased,and Na+ and Cl-were significantly decreased in group HES-E (P＜0.01).Conclusion There is no significant difference in the blood-saving effect between AHH with HES-E and HES-NaCl clinically,but HES-E can maintain homeostasis better.

Objective To investigate the role of CRKL activity in leukemia cells with muhidrug resistance and find new factor related to multidrug resistance. Methods By flow cytometry, CRKL activity was compared in K562, HL-60 cells and its resistance cells. The change of CRKL activity was observed in sensitive cells treated with and withdrawal daunorubicin. Results With the comparison of K562, HL-60 sensitive cells, in K562, HL-60 resistant cell lines, the level of CRKL phosphorylation in K562, HL-60 resistance cells treated with daunombicin 72 hours increased markedly. The level of CRKL phosphorylation was time-dependent with chemotherapy drugs, not change of CRKL activity was found in Jurkat ceils.Conclusion The level of CRKL activity is new factor related to muhidrug resistance in leukemia cells.

Objective To evaluate the protective effect of hydrogen-rich saline on renal ischemia/reperfusion (I/R) in mice. Methods Thirty C57BL/6 mice were randomly divided into 3 groups: sham-operated (SO) group, I/R group (mice were injected with 5 ml/kg saline by tail vein just before ischemia induction) and hydrogen-rich saline group (mice were injected with 5 ml/kg hydrogen-rich saline). At the 6th h after reperfusion, the sera and renal samples subject to IR injury were collected. The Scr and BUN levels in serum were determined and renal histological changes were also examined. The apoptosis of renal tubular epithelial cells was measured by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay. Malondialdehyde (MDA) contents in renal samples were measured using specific kits. The infiltration of F4/80 positive macrophages and neutrophils was assayed by using immunohistochemistry. The mRNA expression of TNF-α, IL-6, IL-1β and IL-17 was detected by using real time reverse transcription PCR. Results As compared with LR group, at the 6th h following reperfusion the levels of Scr and BUN were significantly reduced (P<0.05), histological changes obviously alleviated (P<0.01), apoptosis of renal tubular epithelial cells and MDA contents was decreased (P<0.05) in hydrogen-rich saline group. Moreover, the infiltration of macrophages and neutrophils, and the mRNA expression of TNF-α, IL-6, IL-1β and IL-17 in renal tissue in hydrogen-rich saline group were also declined as compared with IR group (P<0.05). Conclusion Hydrogen-rich saline can ameliorate renal IR injury to some extent, which is associated with inhibition of inflammatory response induced by reperfusion.

To explore the effects of astilbin on the maturation and immunologic function of mouse bone marrow-derived dendritic cells (DCs).

To investigate the protective effects of astilbin on renal ischemia-reperfusion (IR) injury in rats.