Objective · To investigate effect of fucoidan on autophagy, migration and invasion in human multiple myeloma U266 cells. Methods · The U266 cells treated with fucoidan were cultured in vitro. The formation of autophagosomes was observed by transmission electron microscopy (TEM). Transwell assay was used to evaluate the effect of fucoidan on migratory and invasive abilities of U266 cells. The protein levels of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, P62, MMP9, CXCR4, p-AKT/T-AKT, p-mTOR/T-mTOR were detected by Western blotting. MMP9 concentration in the culture medium was examined by ELISA. Results · ① Autophagosomes increased in fucoidan-treated cells compared with control group under TEM. ② Migratory and invasive abilities were inhibited by fucoidan in a dose-dependent manner, which were suppressed by chloroquine. ③ Western blotting demonstrated that expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and MMP9 increased in fucoidan-treated cells, while P62, CXCR4, p-AKT/T-AKT and p-mTOR/T-mTOR decreased compared with control group. ④ The result of ELISA showed that MMP9 concentration in the culture medium of fucoidan-treated cells significantly decreased. Conclusion · Fucoidan induces autophagy and inhibits migration and invasion in U266 cells.

Objective:To establish an experimental model for intracellular antibacteria and endotoxin neutralization in vitro to detect the antibacterial and endotoxin neutralization activity of the muBPI_(25) protein.Methods: RAW264.7 cells were transfected with pcDNA3.1(+)muBPI_(36-259), and then were infected with intracellular bacterial of either G ~+/G~-to establish the experimental model of intracellalar antibacteria.The RAW264.7 cells were co-transfected with the pSecTag2B-muBPI_(36-259) and dual-luciferase reporter gene plasmids for establishment of the experimental model of endotoxin neutralization.Results:The experimental model of intracellular antibacteria confirmed that the muBPI_(25) protein could inhibit/kill Salmonella typhi.The experimental model of endotoxin neutralization indicated that the muBPI_(25) protein could neutralize endotoxin.Conelusion: We firstly demonstrate that murine BPI N-terminal functional fragment(muBPI_(25) protein)can inhibit/kill Salmonella typhi,and can neutralize, its lysating product, endotoxin.

BACKGROUNDSurgery is regarded as the most effective treatment to relieve pain and reduce complications in chronic pancreatitis (CP). Two major strategies exist: duodenum-preserving pancreatic head resection (DPPHR) and pancreatoduodenectomy (PD). Many studies suggest that DPPHR offers advantages during surgery and in the short-term; however, the long-term effects have not been thoroughly investigated. We analyzed the long-term outcomes of DPPHR and PD, over follow-up times of at least 1 year, to determine the optimal surgical treatment for CP.

METHODSWe systemically reviewed all CP surgical treatment reports, and only included randomized controlled trials (RCT) comparing DPPHR and PD, excluding unqualified studies using several pre-specified criteria. When multiple publications of a single trial were found, the most comprehensive current data were selected. Characteristics of the study populations and long-term postoperative outcome parameters were collected. The quality of the studies and data was analyzed using RevMan 4.2 software.

RESULTSFive trials were qualified for meta-analysis, with 261 participants in total (114 in the DPPHR group and 147 in the PD group). There were no significant differences in the age, gender, or indications for surgery of each group. At the mean of 5.7-year (1 - 14 years) follow-up examination, DPPHR and PD resulted in equally effective pain relief, exocrine and endocrine function, and similar mortality rates (P > 0.05); however, DPPHR patients had improved global quality of life and weight gain, and reduced diarrhea and fatigue (P < 0.05).

CONCLUSIONDPPHR and PD result in equal pain relief, mortality, and pancreatic function; however, DPPHR provides superior long-term outcomes.

Duodenum ; surgery ; Humans ; Pancreatectomy ; methods ; Pancreaticoduodenectomy ; methods ; Pancreatitis, Chronic ; psychology ; surgery ; Quality of Life ; Time Factors

OBJECTIVETo modify the current PCR-based method for rapid and efficient preparation of small interfering RNA (siRNA) expression cassette to improve the efficiency of RNA interference.

METHODSThe U6 promoter sequence was amplified by PCR using the genomic DNA of K562 cells as the template, and cloned into pMD18-T vector which served as the template for further PCR amplification with the primers on the plasmid. The amplified product was directly used as the template for preparing siRNA expression cassette. The siRNA expression cassette targeting p53 gene was amplified, verified by sequencing, and transfected into SH-SY5Y cells. After a 48-hour transfection, the cells were harvested and the total RNA was for RT-PCR for evaluating the effect of RNA interference.

RESULTSThe sequencing result confirmed the correct U6 promoter sequence cloned from K562 cells. After transfection of SH-SY5Y cells for 48 h with siRNA expression cassette, the p53 gene expression was inhibited at the mRNA level in comparison with the control cells as demonstrated by RT-PCR detection.

CONCLUSIONThe siRNA expression cassette prepared using the established method described hereby can be well applicable in RNA interference research.

Gene Silencing ; Gene Targeting ; methods ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Polymerase Chain Reaction ; RNA, Small Interfering ; biosynthesis ; genetics ; RNA, Small Nuclear ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

BACKGROUND: Proximal femoral nail anti-rotation is widely used to treat various intertrochanteric fractures. Although its operation trauma is small, and the blood loss of perioperative period is still large. Tranexamic acid has been gradually used to reduce the bleeding of intertrochanteric fracture. The effectiveness and safety of reducing blood loss during perioperative period were not reported. OBJECTIVE: To explore the safety and efficacy of tranexamic acid on perioperative blood loss in patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation. METHODS: One hundred and eight patients with intertrochanteric fracture undergoing proximal femoral nail anti-rotation were selected from First Affiliated Hospital, Guangzhou University of Chinese Medicine between January 2015 and January 2017. Among all the subjects, 52 patients who received the operation before January 2016 served as the control group and 56 patients who received the operation after January 2016 were selected as the treatment group. Half an hour before operation, patients in the treatment group received 1 g tranexamic acid dissolved in 250 mL normal saline by intravenous dropping; patients in the control group just received 250 mL normal saline by intravenous dropping. The bleeding volume, blood transfusion volume, hemoglobin, hematocrit, coagulation index, D-dimer levels and complications were compared between the two groups. RESULTS AND CONCLUSION: (1) During perioperative period, actual blood loss, intraoperative blood loss, dominant blood loss, recessive blood loss, volume of drainage, blood transfusion volume and blood transfusion rate were lower in the treatment group than in the control group (P < 0.05). (2) There was no statistically significant difference in the hemoglobin and hematocrit between the two groups before operation (P > 0.05). The hemoglobin and hematocrit of the two groups gradually decreased after the operation, and there was a slight improvement in the fifth day after surgery. At postoperative 2 hours, 1, 3 and 5 days, the hemoglobin and hematocrit of the treatment group were higher than in the control group (P < 0.05). At preoperation and each time point postoperation, prothrombin time, activated partial thromboplastin time, and fibrinogen levels were not statistically significant between the two groups (P > 0.05). Postoperative D-dimer levels in the two groups were significantly higher than preoperation, and there was a return on the fifth day. There was no statistically significant difference between groups at preoperation and each time point of postoperation (P > 0.05). (3) The results suggest that the tranexamic acid can effectively reduce the dominant and recessive blood loss in patients with the intertrochanteric fracture, and it is safe and effective.

With the utilization of diffusion tensor information of image voxels, a novel MRF (Markov Random Field) segmentation algorithm was proposed for diffusion tensor MRI (DT-MRI) images benefitted from the introduction of Frobenius norm. The comparison of the segmentation effects between the proposed algorithm and K-means segmentation algorithm for DT-MRI image was made, which showed that the new algorithm could segment the DT-MRI images more accurately than the K-means algorithm. Moreover, with the same segmentation algorithm of MRF, better outcomes were achieved in DT-MRI than in conventional MRI (T2WI) image.
Algorithms ; Diffusion Magnetic Resonance Imaging ; methods ; Humans ; Image Interpretation, Computer-Assisted ; methods ; Pattern Recognition, Automated

OBJECTIVETo evaluate the value of spiral computed tomography in the preoperative assessment of the degree of tumor infiltration in mesorectum and circumferential resection margin status of rectal cancer compared with large tissue slice technique.

METHODSFifty-seven patients with rectal cancer diagnosed by fiber colonoscopy and pathology from March 2007 to December 2007 underwent preoperative 64-layers spiral CT examination. The degree of tumor infiltration in mesorectum and circumferential resection margin status were evaluated. Large tissue slice technique was applied in the pathologic study after the total mesorectal excision of the rectum to determine the degree of tumor infiltration in mesorectum and the circumferential resection margin status. The spiral CT findings were compared with pathologic results. The accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the CT results were assessed respectively.

RESULTSThe overall spiral CT accuracy was 93.0% (53/57) for the degree of tumor infiltration in mesorectum, and it was 94.7%, 94.7% and 96.5% for degree I, II, III infiltration, respectively. Fifty-three cases (93.0%) were accurately predicated with the circumferential resection margin status. The sensitivity, specificity, PPV and NPV of spiral CT measurement was 80.0%, 97.6%, 92.3% and 93.2%, respectively, and was consistent well with the histopathological diagnosis.

CONCLUSIONSSpiral CT provides accurate preoperative assessment for the degree of tumor infiltration in mesorectum and circumferential resection margin status of rectal cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Mesentery ; diagnostic imaging ; pathology ; Middle Aged ; Neoplasm Invasiveness ; diagnostic imaging ; Neoplasm Staging ; Predictive Value of Tests ; Rectal Neoplasms ; diagnostic imaging ; pathology ; Rectum ; diagnostic imaging ; pathology ; Sensitivity and Specificity ; Tomography, Spiral Computed

BACKGROUNDDocetaxel (DOC) therapy is well tolerated and shows high response rates in patients with hormone refractory prostate cancer (HRPC). There are many reports on the effect of rapamycin (RPM) on the treatment of carcinogenesis. The goal of this study was to test whether RPM could enhance the susceptibility of both androgen-dependent and -independent prostate carcinoma cells to DOC.

METHODSProstate cancer (PC) cell lines (LNCap, PC3 and AILNCap) were cultured and treated with RPM and DOC alone or in combination. The effects of therapeutic agents on cells were determined by the WST-1 assay. Apoptosis induction was confirmed by flow cytometric analysis. The apopcyto caspase colorimetric assay kit was applied to measure the activities of caspases 3 and 9. The antitumor effects of RPM and DOC against PC cells were also assessed in nude mice using four randomized groups: control, RPM, DOC and combination drug therapy by measuring tumor size. All the animals tolerated both RPM and DOC without significant weight loss.

RESULTSRPM and DOC caused dosage-dependent growth suppression of PC cells. RPM could increase the susceptibility of PC cells to DOC significantly, and combined treatment with RPM and DOC caused synergistic growth suppression in all examined PC cell lines by isobolographic analysis. Both RPM and DOC significantly induced apoptosis in a dosage-dependent manner. RPM (10 nmol/L), DOC (1 nmol/L), and combined treatment induced apoptosis rate were 8%, 17% and 38%, respectively (the control was 2%). RPM could promote the apoptosis induced by DOC in PC cell lines. Both RPM and DOC significantly increased the caspase activity in a dosage-dependent manner. The relative activities of caspase 9 in control, RPM, DOC and RPM + DOC groups were 0.22 +/- 0.02, 0.36 +/- 0.06, 0.47 +/- 0.05 and 0.84 +/- 0.08, respectively. The relative activities of caspase 3 were 0.21 +/- 0.02, 0.24 +/- 0.05, 0.42 +/- 0.06 and 0.81 +/- 0.09, respectively. Either RPM or DOC alone significantly inhibited the growth of PC cells in nude mice compared to the control. The combination of RPM and DOC produced a significant reduction in tumor volume when compared to RPM or DOC alone. After 5-week treatment, the tumor sizes of LNCap in control, RPM, DOC and RPM + DOC groups were (570 +/- 56) mm(3), (412 +/- 41) mm(3), (425 +/- 46) mm(3) and (221 +/- 26) mm(3), respectively.

CONCLUSIONSRPM could significantly increase the susceptibility of both androgen-dependent and -independent PC cells to DOC; the synergy of RPM and DOC was demonstrated. RPM enhanced the DOC-induced upregulation of caspase activity, resulting in an increasing number of cells in sub-G1 phases. The synergy of the combined treatment might be observed in both androgen-dependent and -independent PC cell lines.

Animals ; Antineoplastic Agents ; therapeutic use ; Cell Line, Tumor ; Drug Synergism ; Flow Cytometry ; Humans ; Male ; Mice ; Mice, Nude ; Prostatic Neoplasms ; drug therapy ; Random Allocation ; Sirolimus ; therapeutic use ; Taxoids ; therapeutic use ; Xenograft Model Antitumor Assays

AIMTo establish a sensitive and efficient reporter gene-based screening model for finding agonists of estrogen receptor beta subtype.

METHODSA recombinant vector pTAL-ERE-SEAP was constructed by inserting a synthetic sequence composed of five estrogen responsive elements in front of promoter of pTAL-SEAP vector. pTAL-ERE-SEAP was then transfected into human embryonic kidney (HEK293) cells. G418 (200 microg x mL(-1)) was added to select positive clones that can be induced by E2 to express reporter gene SEAP. The speciality was tested by several ligands of relative nuclear receptors of the same family. The stability of the model, the time-effect relationship, the dose-response relationship, and the immunocytochemistry staining of ERbeta expression after transfection were observed. 2 622 compounds were screened by using this model.

RESULTSStably transfected clones were obtained. The expression levels of reporter gene SEAP of positive clones was induced by E2 in a dose-response and time-effect relationship manners. The Z' factor value was 0.7. The expression levels of dexamethasone and other ligands were low. The result of immunocytochemistry staining showed the expression of ERbeta. E2 had no proliferating effects on stably transfected clones.

CONCLUSIONStably transfected positive clones transfected with recombinant vector pTAL-ERE-SEAP were obtained. The positive clones may be used to screen for agonists of estrogen receptor beta subtype by measurement of luminescent value of expressed SEAP in wells of microlitre plate.

Alkaline Phosphatase ; genetics ; metabolism ; Cell Line ; Estradiol ; pharmacology ; Estrogen Receptor beta ; agonists ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Genetic Vectors ; Humans ; Immunohistochemistry ; Promoter Regions, Genetic ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Response Elements ; genetics ; Transfection

UNLABELLEDTo study the method for dose calculation and beam weight optimization of intensity-modulated radiation therapy (IMRT).

METHODSThe IMRT dose calculation model based on two-dimensional convolution was constructed, the program of dose calculation and beam weight optimization with genetic algorithm was written with Visual c#.Net, and the optimization results were analyzed.

RESULTSGenetic algorithm optimization of beam weights can produce highly conformal dose distributions within a clinically acceptable computation time.

CONCLUSIONGenetic algorithm is valid and efficient in IMRT beam weight optimization, which may facilitate IMRT treatment planning.

Algorithms ; Humans ; Models, Statistical ; Models, Theoretical ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; methods ; Radiotherapy, Conformal ; methods ; standards