Ubiquitin-like proteins are structurally similar to ubiquitin. These proteins are processed, activated, conjugated, and re-leased from conjugates by enzymatic steps that are similar to the corresponding mechanisms for ubiquitin. Ubiquitin-like proteins regu-late a wide array of cellular processes through modification processes, such as nuclear-cytosolic transport, transcriptional regulation, protein stability, response to stress, and progression through the cell cycle. A large number of recent studies have found dysfunctional ubiquitin-like proteins in hepatocellular carcinoma. These proteins are important in tumorigenesis, cell proliferation, apoptosis, and an-giogenesis. Anticancer drug studies revealed that regulating protein modification by using ubiquitin-like proteins may alter the anti-tu-mor effects of chemotherapy and thus influence the chemosensitivity of hepatocellular carcinoma. Results indicate that ubiquitin-like proteins may become a new target for cancer therapy. The mechanism of ubiquitin-like proteins in tumorigenesis and hepatocellular car-cinoma progression is of great significance in the diagnosis and treatment of hepatocellular carcinoma.

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.

Background and purpose: Tumor vasculature is increasingly recognized as a target for cancer therapy. In recent years, a fusion protein consisting of the extra cellular domain of tissue factor (truncated tissue factor, tTF) was fused to the antibody selectively binding to tumor vasculature. Antibody-truncated tissue factor(Ab-tTF) fusion protein specifically induced thrombotic occlusion of tumor vessels resulting in tumor growth retardation or regression in some types of solid tumors. However, there were still some disadvantages in the above approach. We constructed and expressed that the (RGD)_3-tTF fusion protein with peptides arginine-glycine-aspartic acid (GRGDSP, abbr. RGD)as the carrier of tTF to explore whether it bad the capability of targeting to tumor vasculature in the colonic carcinoma model. Methods: The (RGD)_3-tTF fusion gene consisting of the tTF was fused to three series-wound peptides RGD. The (RGD)_3-tTF construct was expressed in Escherichia coil BL21(DE_3). The fusion protein was purified through Nickel affinity chromatography column. The activity of inducing blood coagulation was detected by clotting assay and coagulation factor X (FX) activation assay. The specific binding to integrins α_vβ_3 was analyzed by indirect enzyme linked immunosorbent assay (ELISA). All these were compared with the fusion protein RGD-tTE Colonic nude mice models were randomly divided into 3 groups (1 nude mice per group).Tumors were stained by the (RGD)_3-tTE RGD-tTF fusion protein and tTF which were labeled with Fluorescein Isothiocyanate(FITC). The location of the (RGD)_3-tTF fusion protein in the colonic carcinoma bearing nude mice tissue was analyzed by immunofluorescence assay. Results: The (RGD)_3-tTF fusion protein retained tissue factor thrombogenic activities. With increasing concentration, the clotting time was shortened correspondingly. Under the conditions of Ca~(2+), the clotting time was 9.96±0.56 min when the concentration was 6 μmol/L(P<0.01). The (RGD)_3-tTF fusion protein could activise F X above 6 μmol/L concentration, which was similar to RGD-tTF fusion (F=0.147, P>0.05). The ability of the (RGD)_3-tTF fusion protein binding specifically to integrins α_vβ_3 was stronger than that of the RGD-tTF fusion protein in the same concentration (F=164.81, P<0.01), which was apparently indicated by the A_(405nm) 1.25 and 0.95 when the concentration was 0.24 μmol/L. Immunofluorescence assay showed that the (RGD)_3-tTF fusion protein was assembling in the tumor vasculature of the colonic carcinoma bearing nude mice. Conclusion: The (RGD)_3-tTF fusion protein which retained tissue factor thrombogenic activities could bind specifically and efficiently to tumor vasculature in the colonic carcinoma bearing mice through binding to the tumor marker integrins α_vβ_3. It might be a promising foundation for further studies on the colon cancer molecular targeted therapy with tTF as an effective factor.

Ischemic stroke is an important cause of death and disability.Hemorrhagic transformation (HT) is a serious complication of acute ischemic stroke,especially in patients receiving intravenous thrombolytic therapy.Identifying patients at high risk of HT is very important for improving stroke outcomes.Some studied have shown that several serum biomarkers can be used to predict HT risk in patients with acute ischemic stroke.This article reviews the serum biomarkers of HT after acute ischemic stroke.

Background and purpose: In recent years, more and more clinical researches on the neuroendocrine carcinoma of the breast were carried out at home and abroad. Although there are quite a lot of the retrieved documents of NEBC at home and abroad, but large-scale reports are rare, besides, the epidemiology, diagnosis, treatment and prognosis were different;more research are needed to analyze NEBC. This paper mainly discussed the NEBC clinical diagnosis, treatment and prognosis. Methods: A retrospective analysis was carried out, 25 cases of Tumor Hospital Afifliated to Xinjiang Medical University from Jan. 2004 to Jun. 2013 were pathologically diagnosed as NEBC by clinical data and the follow-up. Results:The average age of 25 NEBC patients was 58.2 years old, without clinical and imaging characteristic features, immunohistochemistry staining showed that, the estrogen receptor (ER) and progesterone receptor (PR) positive rates were 76%and 64%. No one showed HER-2 strong positive. The follow-up was 9-115 months. Besides, 1-, 2-and 5-year overall survival (OS) rates were 100%, 95%and 88%, the disease-free survival (DFS) rates were 96%, 90%and 78%. Conclusion:The age of the patients with NEBC in this study was lower than the data abroad. Age, tumor size, pathological staging may be related to the prognosis of NEBC, and postoperative comprehensive treatment options need further study.

OBJECTIVE:To investigate the status quo of teachers'teaching ability of standardized training for hospital pharma-cists in Beijing area,and provide reference for strengthening the faculty construction. METHODS:A dynamic assessment of stan-dardized training base for hospital pharmacists of 17 inpatients in Beijing area was conducted to organize evaluation experts to as-sess the teaching ability of outpatient pharmacy,ward pharmacy and clinical pharmacy in training base. Questionnaire was designed and trainees were organized to evaluate the teaching ability,time,attitude,feedback and others and related results of teaching in bases were vegarded as objects for analysis and summary. RESULTS:The overall ability of teaching was high,and the most promi-nent was clinical pharmacy,which was well received by trainees. But there are differences among departments,the ability of ward pharmacy is slightly lower than the clinical pharmacy and outpatient pharmacy. And the teaching ability still had great improvement. CONCLUSIONS:In order to improve the overall teacher strength in training base,competence-based training for hospital pharma-cists must be strengthened to encourage teachers to attend training courses,strengthen cultivation of humanistic quality and enrich students'evaluation forms.

OBJECTIVETo compare the clinical effective differences between acupuncture-moxibustion and Chinese herbs for functional dyspepsia with emotional disorder.

METHODSEighty patients were randomly divided into an observation group and a control group, 40 cases in each one. In the observation group, based on the basic treatment Zusanli (ST 36), Yinlingquan (SP 9), Zhongwan (CV 12), Danzhong (CV 17), Neiguan (PC 6), Taichong (LR 3), Ganshu (BL 18), Geshu (BL 17) and Danshu (BL 19) were selected once every day or every other day. The treatment was given 14 times. In the control group, based on the basic treatment individual therapy of Chinese herbs according to syndrome differentiation was applied,one dose a day. Patients were treated for 4 weeks in the two groups. The scores of self-rating depression scale (SDS), the main symptoms and satisfied degree by self-rating for treatment were observed before and after 4-week treatment in the two groups. Also, the rate and the average time of return visit were compared in 6 months after treatment. Results After treatment, the SDS scores and the main symptoms scores were all improved compared with those before treatment in the two groups (all P< 0. 05). After 4-week treatment, the improvement of the SDS score in the observation group was better than that in the control group (48. 9±8. 5 vs 53. 1±8. 0, P<0. 05). After 1-week treatment and 4-week treatment, the improvements of the main symptoms in the observation group were superior to those in the control group (2. 7 ± 1. 0 vs 3. 3±0. 9, 1. 5±0. 9 vs 2. 3±1. 1, both P<0. 05), and the satisfied degree scores by self-rating were better than those in the control group (3. 0±1. 1 vs 3. 8±1. 3, 2. 0±1. 4 vs 2. 9±1. 5, both P<0. 05). In 6 months after treatment,the return visit rate in the observation group was 42. 5% (17/40), and it was lower than 70. 0% (28/40) in the control group (P<0. 05). The average time of return visit in the observation group was less than that in the control group (1. 0±0.8 vs 1. 9±0. 7, P<0. 05).

CONCLUSIONThe effect of acupuncture-moxibustion or functional dyspepsia with emotional disorder is better than that of Chinese medicine.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Dyspepsia ; psychology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome

Objective To synthesize 131I labeled anti-neuropilin-1 monoclonal antibody A6 (131IA6) and evaluate its biodistribution and imaging in malignant glioma xenografts.Methods (1) A6 was labeled with 131I by Iodogen method under the optimum labeling conditions,then the labeling efficiency,radiochemical purity and stability were measured in vitro.(2) In vitro bioactivity,cellular uptake and receptor affinity of 131I-A6 with U87MG cells were measured.(3) The nude mice bearing human U87MG cells were randomly divided into 5 groups with 5 in each group.The nude mice were sacrificed by cervical dislocation and dissected at 24,48,72,96,and 120 h,respectively,after intravenous injection of 1.2 MBq 131I-A6.The biodistribution of the agent was measured as ％ID/g,and the ratios of tumor/blood (T/B) and tumor/muscle (T/M) were calculated.(4) SPECT/CT imaging was performed in 6 mice including 3 in the competitive inhibition control group at 24,48,72,96,and 120 h post injection.Two-sample t test was used for data analysis.Results (1) The labeling yield of 131I-A6 was (95.46±3.34)％,and the radiochemical purity was more than 95％.At 96 h of incubation in PBS,the radiochemical purity was more than 85％.(2)131I-A6 had rapid accumulation in U87MG cells and reached the peak of (15.80±1.30)％ at 1 h.When the probe was incubated with large excesses of non-radioactive A6,the uptake level of 131I-A6 in U87MG cells was significantly inhibited (t=2.862,P＜0.05).Kd of 131I-A6 binding to NRP-1 was (1.67±0.14) nmol/L in U87MG cells.(3) Biodistribution study showed that the uptake in blood,liver and tumor was (8.00±1.42),(7.68±1.56) and (6.00±1.24) ％ID/g at 24 h,respectively.The uptake in muscle,brain and bone was lower.The T/B and T/M were 0.78±0.10 and 3.20±0.30 at 24 h,and they reached the highest level of 1.87±0.50 and 7.13±0.24 at 120 h.(4) The SPECT imaging showed that the tumors could be visualized at 24 h and delineated more clearly at 120 h post injection of 131I-A6.Conclusions 131I-A6 can be easily synthesized by Iodogen method with high radiochemical purity.The specific tumor uptake of 131I-A6,which correlates with NRP-1 expression in gliomas,suggests that it may be a new promising tumor targeting radiotracer.

Objective To evaluate the effect of 15-deoxy-△12,14-prostaglandin J2 (15d-PGJ2) on endotoxin-induced acute lung injury (ALI) in rats.Methods Forty healthy male Sprague-Dawley rats, aged 3-5 months, weighing 220-250 g, were randomly divided into 4 groups (n =10 each) using a random number table: control group (group C), 15d-PGJ2 group, lipopolysaccharide (LPS) group, and LPS +15d-PGJ2 group.In group 15d-PGJ2, 15d-PGJ20.3 mg/kg was injected via the tail vein, while the equal volume of normal saline was given in group C.In LPS and LPS+15d-PGJ2 groups, ALI was produced with LPS 6 mg/kg injected through the tail vein, and then the equal volume of normal saline and 15d-PGJ2 0.3 mg/kg were injected, respectively.At 4 h after LPS injection, blood samples were drawn from the abdominal aorta for blood gas analysis, and arterial oxygen partial pressure (PaO2) was recorded.The rats were then sacrificed, lungs were removed for microscopic examination, and for determination of wet/dry lung weight ratio (W/D ratio), TNF-α, IL-8 and cytokine-induced neutrophil chemoattractant-1 (CINC-1) contents (by enzyme-linked immunosorbent assay) , and nuclear factor kappa B (NF-κB) p65 and IκB-α expression (by Western blot).Results Compared with group C, no significant change was found in PaO2, W/D ratio, contents of TNF-α, IL-8 and CINC-1, and expression of NF-κB p65 and IκB-α in group 15d-PGJ2 (P＞0.05), and PaO2 was significantly decreased, W/D ratio and contents of TNF-α,IL-8 and CINC-1 were increased, the expression of NF-κB p65 was up-regulated, and the expression of IκB-α was down-regulated in LPS and LPS+ 15d-PGJ2 groups (P＜0.05).Compared with group LPS,PaO2 was significantly increased, W/D ratio and contents of TNF-α, IL-8 and CINC-1 were decreased, the expression of NF-κB p65 was down-regulated, and the expression of IκB-α was up-regulated (P＜0.05),and the pathological changes were attenuated in group LPS+ 15d-PGJ2.Conclusion 15d-PGJ2 can mitigate endotoxin-induced ALI in rats.

Objective To investigate the cell types of renal allograft glomerulitis in posttransplantation patients and its relationship with peritubular capillary (PTC) C4d deposition.Methods Fifty-one allograft kidney transplant recipients diagnosed as acute rejection from June 2006 to June 2009 in our center were enrolled in this retrospective study. Fifty-one biopsy specimens of these patients accompanied with glomerulitis were examined by immunostain for macrophages, T cells, granzyme B, Foxp3 and C4d. Glomeular macrophages, T cells, cytotoxic T cells, regulatory T cells were counted. The recipients were divided into C4d+ and C4d- groups according to the C4d deposition in peritubular capillaries. Results The total number of glomerular infiltrating cells in C4d+ group was significantly higher than that in C4d- group (17.79±7.70 vs 8.17±3.80, P＜0.01). The infiltrating cell was predominantly macrophage in CAd+ group and significantly higher as compared to C4d- group (13.73 ±7.03 vs 2.57 ±1.22, P＜0.01).However, the infiltrating cell was predominantly T cell in C4d- group and significantly higher as compared to C4d+ group (5.60±2.81 vs 4.05±2.60, P=0.023). The infiltrating T cells in both groups were predominantly cytotoxic T cells. Conclusions The total number of infiltrating cells in glomerulitis is related to peritubular capillary C4d deposition. The infiltrating cell is predominantly macrophage in C4d+ group and predominantly T cell in C4d- group. Meanwhile the infiltrating T cell in both groups is predominantly cytotoxic T cell.