Decreased expression of Sushi Domain Containing 2 correlates to progressive features in patients with hepatocellular carcinoma.

BACKGROUND: Sushi Domain Containing 2 (SUSD2) has been identified as a regulator of colon and breast cancer. Increasing evidence suggests that SUSD2 plays a key role in tumorigenesis. However, the SUSD2 expression status and its functions in hepatocellular carcinoma (HCC) are still unrevealed. In the present study, we intended to investigate SUSD2 expression status and its correlation with the clinicopathological features in HCC patients. Furthermore,we examined the influence of SUSD2 on the proliferation, apoptosis, invasion and migration of the HCC cell lines HepG2 and SMMC7721. METHODS: We evaluated the SUSD2 expression in HCC tissues and paired normal liver tissues by quantitative real-time PCR and western blotting analysis. The clinicopathological significance of SUSD2 was investigated by immunohistochemistry (IHC) on a HCC tissue microarray. Receiver operating characteristic (ROC) analysis was applied to determine the optimal cut-off score for positive expression of SUSD2. The correlation between SUSD2 protein expression and clinicopathological features of HCC was analyzed by Chi square test. The cell proliferation, apoptosis, invasion and migration potential were observed to detect the functions of SUSD2 in HCC cells. RESULTS: Decreased expression of SUSD2 mRNA and protein were observed in the majority of HCC tissues, compared with paired normal liver tissues. When SUSD2 high expression percentage was determined to be above 52.5 % (area under ROC curve = 0.769, P = 0.000), low expression of SUSD2 was observed in 62.2 % (112/180) of HCC tissues and high expression of SUSD2 was observed in all normal liver tissues (16/16) by IHC. Decreased expression of SUSD2 in patients was correlated with high histological grade (χ(2) = 5.198, P = 0.023), advanced clinical stage (χ(2) = 30.244, P = 0.000), pT status (χ(2) = 33.175, P = 0.000), pN status (χ(2) = 4.785, P = 0.029), pM status (χ(2) = 4.620, P = 0.032). Down-regulation of SUSD2 promoted cell proliferation,invasion and migration,reduced the cell apoptosis. CONCLUSIONS: Our findings suggest that SUSD2 may play as a tumor suppressor in HCC cells and could be served as an additional potential marker for diagnosis.

Xylomolins O-X: New limonoids from the Thai mangrove Xylocarpus moluccensis.

Ten new limonoids, named xylomolins O-X, were isolated from seeds of the mangrove Xylocarpus moluccensis, collected in the mangrove swamp of Trang Province, Thailand. Their structures were elucidated on the basis of comprehensive spectroscopic data analysis. The absolute configurations of five compounds (1, 3, 8-10) were unequivocally determined by single-crystal X-ray diffraction analyses, conducted with Cu Kα radiation. Xylomolins OU (1-7) are structurally intriguing mexicanolides, and xylomolin V (8) is a derivative of azadirone. Xylomolin W (9) is the first phragmalin 1,8,9-orthoester with report on X-ray crystallography from the genus Xylocarpus. In addition, xylomolin X (10) is the fifth member of the khayalactone class of limonoids with a hexahydro-2H-2,5-propanocyclopenta[b]furan motif. Compounds 1-10 inhibited NO production in LPS-activated RAW 264.7 macrophages in the range of 10.45-95.47% at the concentration of 100.0 µM. Xylomolin X (10) and xylomolin V (8), exhibited the most potent activity with IC50 values of 9.90 ± 1.84 µM and 14.66 ± 2.33 µM, respectively.

[Effect of ribavirin upon secretion of thymic stromal lymphopoietin in respiratory syncytial virus-induced asthma exacerbation of mice].

OBJECTIVE: To investigate the role of ribavirin (RIB) in treating respiratory syncytial virus (RSV)-induced asthma exacerbation of mice. METHODS: (1) Cell experiment: 32 flasks of human airway epithelial cell 16HBEs were randomly divided into four groups: RSV group, RSV/RIB group, RIB group and control group. 16HBEs were infected with RSV at a multiplicity of infection (MOI) of 2. RIB 50 microg/ml was added in culture medium and Western blot used to detect the production of thymic stromal lymphopoietin (TSLP) protein; (2) Animal experiment: 32 female BALB/c mice were randomly divided into four groups: Ovalbumin (OVA) group, OVA/RSV group, OVA/RSV/RIB group and control group. Mice were sensitized by OVA and stimulated with nebulized OVA. RSV was inoculated into murine nasal cavity and RIB 10 mg/kg intramuscularly administered. BUXCO noninvasive murine lung function detection instrument was used to examine the airway response to metacholine; ELISA was used to detect IL-4, IL-5, IL-13 and IFN-gamma in murine serum and TSLP in supernatants of bronchoalveolar lavage fluid (BALF); murine lung specimens were stained with HE to observe inflammation and immunohistochemical technique was employed to observe the production of TSLP in murine airway epithelial cells. RESULTS: The cell experiment demonstrated the productions of TSLP protein in RSV group, RSV/RIB group, RIB group and control group were 1.97 +/- 0.22, 1.16 +/- 0.19, 0.99 +/- 0.17 and 0.89 +/- 0.08 respectively, and the production of TSLP in RSV/RIB group was lower than that in RSV group (P < 0.01). The animal experiment demonstrated that the murine airway responsiveness in RSV/OVA/RIB group was lower than that in OVA/RSV group (P < 0.01). The levels of IL-4 [(109.7 +/- 41.9) pg/ml], IL-5 [(220.8 +/- 30.9) pg/ml], IFN-gamma [(13.0 +/- 3.4) pg/ml] in murine serum and TSLP [(1945 +/- 82) pg/ml] in BALF of RSV/OVA/RIB group were significantly lower than those in OVA/RSV group [(274.2 +/- 103.7), (293.3 +/- 46.1), (30.1 +/- 5.7) and (2127 +/- 46) pg/ml respectively, all P < 0.01]; less infiltration of airway inflammatory cells in OVA/RSV/RIB group was observed than that in OVA/RSV group. Immunohistochemical staining of TSLP also showed a lower production of TSLP in airway epithelial cells of OVA/RSV/RIB group than OVA/RSV group. CONCLUSION: Ribavirin can inhibit the elevated production of TSLP after RSV infection and relieve RSV-induced asthma exacerbation in mice.

[Effect of adjusting the immunosuppressants on the prognosis of the infection in the patients with liver transplantation].

OBJECTIVE: To explore the method of adjusting the immunosuppressants in serious infection after liver transplantation. METHODS: With reference to sepsis-related organ failure assessment (SOFA), 2005.1-2007.12, when the patient's score > or =15, the immunosuppressants were withdrawn, and the patients were given powerful antibiotics and the other treatments in combination. They were further divided into two groups, SOFA 15-17 (group A, 10 cases) and > or =18 (group B, 16 cases). They were compared, and also with the patients without stoppage of immunosuppressants (group C, 13 cases, 2003.3-2004.12). After withdrawing the immunosuppressant, the rejection incidence and times, the changes in SOFA score and mortality and their relationships were analyzed. RESULTS: After adjusting the immunosuppressant and with control of serious infections, rejection occurred in 9 patients, with 5 cases in group A (50.0%), 4 in B (25.0%), none in C. The differences among groups showed statistically significant difference (chi(2)=8.0, P=0.02), but no difference was seen between group A and B (chi(2)=1.70, P=0.19). When the rejection developed, the SOFA score decreased obviously (9.78+/-3.14 vs. 17.22+/-1.86, t=6.10, P=0.00). The time of rejection was (17.56+/-2.60) days after stopping the immunosuppressant. All 25 deaths were due to serious infection with multiple organ dysfunction syndrome, but not rejection. Five deaths occurred in group A (50.0%), 7 in B (43.8%), 13 in C (100.0%). Not a single patient with rejection died from infection. Proper adjustment of the immunosuppressants could decrease the mortality (chi(2)=7.60, P=0.02). CONCLUSION: SOFA score could be used to guide the adjustment of the immunosuppressants, when SOFA> or =15, the immunosuppressants could be stopped, which would not increase the rejection incidence and decrease mortality. The lower the SOFA score is, the faster the patients recuperate better, but more rejection develops. In order to adjust the immunosuppressant in time, the period with high SOFA score should be shortened.

[Non-rigid medical image registration based on mutual information and thin-plate spline].

To get precise and complete details, the contrast in different images is needed in medical diagnosis and computer assisted treatment. The image registration is the basis of contrast, but the regular rigid registration does not satisfy the clinic requirements. A non-rigid medical image registration method based on mutual information and thin-plate spline was present. Firstly, registering two images globally based on mutual information; secondly, dividing reference image and global-registered image into blocks and registering them; then getting the thin-plate spline transformation according to the shift of blocks' center; finally, applying the transformation to the global-registered image. The results show that the method is more precise than the global rigid registration based on mutual information and it reduces the complexity of getting control points and satisfy the clinic requirements better by getting control points of the thin-plate transformation automatically.

Urotensin II promotes aldosterone expression in rat aortic adventitial fibroblasts.

Urotensin II (UII) contributes to cardiovascular diseases by activating vasoactive peptides. The present study aimed to determine the effect of UII on aldosterone (ALD) and its receptor in cultured adventitial fibroblasts (AFs) and the tunica adventitia of rat vessels to explore the possible mechanisms underlying vascular remodeling. Expression levels of aldosterone and its receptor on tunica adventitia were determined using immunohistochemistry. Growth­arrested AFs and tunica adventitia from rat vessels were incubated with UII and inhibitors of various signal transduction pathways. ALD receptor (ALD­R) mRNA expression levels and ALD protein exoression levels were determined by reverse transcription­quantitative polymerase chain reaction and ELISA, respectively. Aldosterone and its receptors were expressed on tunica adventitia. UII promoted ALD protein secretion from cells in a dose­ and time­dependent manner. ALD­R mRNA expression in cells was also dysregulated. Furthermore, the effects of UII were substantially inhibited by treatment with the inhibitors PD98059, Y­27632, H­7, CSA and nicardipine. These results were further verified in the tunica adventitia of rat vessels. The present findings indicated that UII stimulated ALD protein secretion and ALD­R mRNA expression in AFs and in the tunica adventitia of rat vessels; moreover, this effect may be mediated by signal transduction pathways involving MAPK, Rho, PKC, calcineurin and Ca2+. UII may also contribute to vascular remodeling by stimulating the production of ALD and its receptor.

[Development of a three-dimensional precise conformal radiotherapy treatment planning system based on a cobalt-60 teletherapy unit].

In this paper, a three-dimensional precise conformal radiotherapy treatment planning system based on a cobalt-60 teletherapy unit is introduced. With the help of additional precise target localization and conformal field-shaping devices, the TPS can greatly improve the performance of conventional cobalt-60 teletherapy units in precise target localization, radiotherapy planning and dose delivery. The clinical practices show that the TPS has advantages of reliable precision and an affordable price , and it is urgently needed in our country.

[Optimization methods in radiation treatment planning].

The optimization methods in radiation treatment planning are reviewed in this paper, including the physical and biological optimization models, the optimization for Gamma knife treatment planning, the optimization for intensity modulated radiation treatment planning and the optimization for intravascular brachytherapy treatment planning. The development trend of radiation treatment planning is also introduced in the paper.

The association between aquaporin-1 expression, microvessel density and the clinicopathological features of hepatocellular carcinoma.

The present study aimed to evaluate the effects of aquaporin-1 (AQP1) level and intratumoral microvessel density (IMD) on the clinicopathological features of patients with hepatocellular carcinoma (HCC). The AQP1 expression levels, IMD and AQP1/IMD ratios in patients with HCC were measured using a semi-quantitative immunohistochemical technique. The association between these features and clinicopathological variables were evaluated. The prognostic impact of AQP1 and IMD on overall survival (OS), and 5-year disease-free survival (DFS) of HCC patients was investigated retrospectively. P<0.05 was considered to indicate a statistically significant difference. A total of 90 cases of HCC were included in the present study. AQP1 was markedly expressed in the membranes of microvessels and small vessels, but seldom in hepatocellular carcinoma cells. Blood vessels in the tumors were markedly stained by anti-cluster of differentiation 34 antibody. AQP1 expression and IMD was significantly correlated with tumor size, histologic grade, Child-Pugh classification, microvascular invasion and tumor-node-metastasis (TNM) stage (P<0.05). Concurrently, for the 5-year DFS and OS, a larger tumor size, poorly differentiated histological grade, B and C Child-Pugh classification, presence of microvascular invasion, high TNM stage, a high AQP1 expression and a high IMD were significant risk factors for mortality. Multivariate analysis revealed that TNM stage and IMD were independent unfavorable prognostic markers for 5-year DFS (P=0.049 and P=0.025, respectively) and OS (P=0.043 and P=0.042, respectively). These data suggest that high AQP1 expression and IMD are associated with tumor progression and prognosis in HCC. The IMD level may serve as an independent indicator for the 5-year DFS and OS.

[A review of methods for correction of artifacts in functional MRI].

There are many kinds of artifacts in image series of functional MRI, such as head motion artifacts, physiological motion artifacts, blood flow artifacts, ghost artifacts and susceptibility artifacts. These artifacts which are unassociated with the neural activities, have so severe affects on the analysis of functional MRI data that they not only reduce the sensibility and reliability of functional MRI, but also make the detecting, locating and visualizing of the functional active regions more complicated. The mechanism and the effect of artifacts in functional MRI are discussed here, the methods of correcting artifacts are reviewed, and research prospects are discussed too.

Does ligand-receptor mediated competitive effect or penetrating effect of iRGD peptide when co-administration with iRGD-modified SSL?

Ligand-mediated targeting of anticancer therapeutic agents is a useful strategy for improving anti-tumor efficacy. It has been reported that co-administration of a tumor-penetrating peptide iRGD (CRGDK/RGPD/EC) enhances the efficacy of anticancer drugs. Here, we designed an experiment involving co-administration of iRGD-SSL-DOX with free iRGD to B16-F10 tumor bearing mice to examine the action of free iRGD. We also designed an experiment to investigate the location of iRGD-modified SSL when co-administered with free iRGD or free RGD to B16-F10 tumor bearing nude mice. Considering the sequence of iRGD, we selected the GPDC, RGD and CRGDK as targeting ligands to investigate the targeting effect of these peptides compared with iRGD on B16-F10 and MCF-7 cells, with or without enzymatic degradation. Finally, we selected free RGD, free CRGDK and free iRGD as ligand to investigate the inhibitory effect on RGD-, CRGDK- or iRGD-modified SSL on B16-F10 or MCF-7 cells. Our results indicated that iRGD targeting to tumor cells was ligand-receptor mediated involving RGD to αv-integrin receptor and CRGDK to NRP-1 receptor. Being competitive effect, the administration of free iRGD would not be able to further enhance the anti-tumor activity of iRGD-modified SSL. There is no need to co-administrate of free iRGD with the iRGD-modified nanoparticles for further therapeutic benefit.

[Inhibitory effects of endostatin on interleukin-6 and interleukin-8 release by human umbilical vein endothelial cells].

OBJECTIVE: To investigate the inhibitory effects exercised by endostatin on the production of interleukin-6 (IL-6) and IL-8 by human umbilical vein endothelial cells (HUVECs). METHODS: (HUVECs were isolated and cultured in vitro with endostatin (treated group) or PBS (control group), and the supernatant was harvested from the primary culture medium daily for 9 consecutive days starting from the first day of culture, followed by centrifugation. IL-6 and IL-8 contents in the supernatant were measured using sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: IL-6 and IL-8 were detected in the supernatant of the control cell culture, and their amounts increased as the cell culture was prolonged, reaching the peak levels on day 6 (2 979.32+/-19.65 pg/ml and 6 018.87+/-56.74 pg/ml, respectively). In the treated group, however, the amounts of IL-6 and IL-8 were significantly lower than the control levels (P<0.01). CONCLUSION: Endostatin can inhibit the growth and proliferation of endothelial cells, reducing their biological activities.

[Inhibitory effect of recombinant human endostatin on angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795].

OBJECTIVE: To study the inhibitory effect of recombinant human endostatin (rhES) on the angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795. METHODS: The recombinant yeast strain containing the gene sequence encoding highly soluble rhES was induced by methanol for rhES production, which was purified with heparin affinity chromatography. T739 mice with subcutaneous inoculation of LA795 cells were randomized into 2 groups (10 in each group) to receive injection of either rhES (20 mg/kg x b x w x per day) or PBS in the same volume for 14 consecutive days starting from the sixth day after the inoculation. The angiogenesis and lung metastasis of the implanted tumors were subsequently observed. RESULTS: Purified rhES was successfully obtained. As shown by immunohistochemistry, the tumors in the mice receiving rhES treatment exhibited less density of the microvessels than those in the PBS-treated mice did (P<0.01). Pathological examination of the lung tissue of the mice in rhES group found no visible signs of tumor metastasis, which, in contrast, was widespread in PBS group. The weight of the lungs was also significantly different (P<0.01). CONCLUSION: rhES possesses good biological properties and can potently inhibit the angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795.

[Synthesis, cloning, expression and antigenicity of the therapeutic multi-epitope gene of hepatitis B virus and of its products].

OBJECTIVE: To study the synthesis, cloning, expression and antigenicity of therapeutic multi-epitope gene of hepatitis B virus. METHODS: The therapeutic multi-epitope gene of hepatitis B virus was synthesized and cloned into the vector pWR450-1, then was expressed in E.coli and the products were purified. The immunogenicity of the expressed protein was analyzed by Western-blotting. RESULTS: Recombinant plasmid PWR/BPT was constructed successfully and the protein of multi-epitope gene of hepatitis B virus was expressed in E. coli, which showed ideal antigenicity by Western-blotting. CONCLUSIONS: The designed multi-epitope therapeutic gene of hepatitis B virus was proved to be correct and the expressed protein may be a good therapeutic vaccine.

[Virtual imagery in medicine].

The development of modern medical imaging and related technologies, and the increasing requirements for accuracy in clinical diagnosis and treatments give the emergence of a new discipline--virtual imagery in medicine. In this paper, we try to summarize its technical aspects, including virtual reality, 3-D image reconstruction and visualization, virtual endoscopy, and so on.

[DICOM in radiotherapy].

Digital Image Communication in Medicine (DICOM) defines a standard method to store and transmit digital medical image information, in which there is a piece of implemented protocol named DICOM-RT that specially addresses both the transmission of radiation therapy image data and the ancillary data. In this paper, we firstly introduce the DICOM-RT with the emphases on its components, relationship with radiotherapy and how to produce the DICOM-RT object that refer to some certain radiotherapy information. Then we expatiate on the impact that benefits from applying DICOM-RT to radiotherapy, with an aid to accelerate its application in China.

[The 3D PACS image system based on pipeline framework].

We put forward an image rendering system based on pipeline framework for processing and displaying medical images. Compared to original computer graphics algorithms divided into volume rendering and surface rendering, this framework can effectively comprehend methods of computer graphics and image processing, import some new concepts such as vertex buffer, pixel buffer and texture buffer. We implement Shaded Surface Display, Maximum Intensity Projection, Digitally Reconstructed Radiography, Multi planar Reformation, Curved Planar Reformation and Interactive Virtual Endoscopy in our new developed PACS image system.

Silencing of periostin inhibits nicotine-mediated tumor cell growth and epithelial-mesenchymal transition in lung cancer cells.

Nicotine has been found to induce the proliferation of lung cancer cells through tumor invasion and to confer resistance to apoptosis. Periostin is abnormally highly expressed in lung cancer and is correlated with angiogenesis, invasion and metastasis. Here, we investigated the roles of periostin in the lung cancer cell proliferation, drug resistance, invasion and epithelial-mesenchymal transition (EMT) induced by nicotine. The periostin gene was silenced using small interfering RNA (siRNA) in A549 non-small cell lung cancer (NSCLC) cells. The cells were transfected with control or periostin siRNA plasmids. Periostin mRNA was evaluated by quantitative reverse transcription-polymerase chain reaction (RT-PCR). Cell proliferation was detected using the MTT assay and cell apoptosis was detected by Annexin V-FITC and propidium iodide (PI) double staining. Tumor invasion was detected by the Boyden chamber invasion assay. Western blotting was performed to detect the expression of the EMT marker Snail. Our results revealed that stably periostin-silenced cells were acquired by G418 screening, and the periostin mRNA expression levels of which were decreased by nearly 80%. Periostin-silenced A549 cells exhibited reduced cell proliferation, elevated sensitivity to chemotherapy with cisplatin, decreased cell invasion and Snail expression (P<0.05). Nicotine upregulated the periostin protein levels in the A549 cells and this upregulation was not blocked by the generalized nicotinic acetylcholine receptor (nAChR) antagonist, hexamethonium. In conclusion, periostin is one of the targets regulated by nicotine in lung cancer cells and is involved in the cancer cell growth, drug resistance, invasion and EMT induced by nicotine.

The antitumor activity of a doxorubicin loaded, iRGD-modified sterically-stabilized liposome on B16-F10 melanoma cells: in vitro and in vivo evaluation.

Considering the fact that iRGD (tumor-homing peptide) demonstrates tumor-targeting and tumor-penetrating activity, and that B16-F10 (murine melanoma) cells overexpress both αv integrin receptor and neuropilin-1 (NRP-1), the purpose of this study was to prepare a novel doxorubicin (DOX)-loaded, iRGD-modified, sterically-stabilized liposome (SSL) (iRGD-SSL-DOX) in order to evaluate its antitumor activity on B16-F10 melanoma cells in vitro and in vivo. The iRGD-SSL-DOX was prepared using a thin-film hydration method. The characteristics of iRGD-SSL-DOX were evaluated. The in vitro leakage of DOX from iRGD-SSL-DOX was tested. The in vitro tumor-targeting and tumor-penetrating characteristics of iRGD-modified liposomes on B16-F10 cells were investigated. The in vivo tumor-targeting and tumor-penetrating activities of iRGD-modified liposomes were performed in B16-F10 tumor-bearing nude mice. The antitumor effect of iRGD-SSL-DOX was evaluated in B16-F10 tumor-bearing C57BL/6 mice in vivo. The average particle size of the iRGD-SSL-DOX was found to be 91 nm with a polydispersity index (PDI) of 0.16. The entrapment efficiency of iRGD-SSL-DOX was 98.36%. The leakage of DOX from iRGD-SSL-DOX at the 24-hour time point was only 7.5%. The results obtained from the in vitro flow cytometry and confocal microscopy, as well as in vivo biodistribution and confocal immunofluorescence microscopy experiments, indicate that the tumor-targeting and tumor-penetrating activity of the iRGD-modified SSL was higher than that of unmodified SSL. In vivo antitumor activity results showed that the antitumor effect of iRGD-SSL-DOX against melanoma tumors was higher than that of SSL-DOX in B16-F10 tumor-bearing mice. In conclusion, the iRGD-SSL-DOX is a tumor-targeting and tumor-penetrating peptide modified liposome which has significant antitumor activity against melanoma tumors.