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.

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.

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.

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.

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.

Anti-tumor and anti-angiogenic effect of metronomic cyclic NGR-modified liposomes containing paclitaxel.

In the present study, we prepared NGR-modified sterically stabilized liposomes containing paclitaxel (NGR-SSL-PTX) in order to evaluate their potential targeting to aminopeptidase N receptors expressed on tumor endothelial cells and the tumor cell surface and its anti-angiogenic activity following metronomic administration. NGR-SSL-PTX was prepared by a thin-film hydration method. The in vitro targeting characteristics of NGR-modified liposomes on HUVEC (human umbilical vein endothelial cells), HT1080 (human fibrosarcoma cells) and MCF-7 (human breast adenocarcinoma cells) were then investigated. The effect of NGR-SSL-PTX on HUVEC proliferation and migration was also tested. The pharmacokinetics of NGR-SSL-PTX was studied in rats. The in vivo targeting activity of NGR-modified liposomes was investigated in HT1080 tumor-bearing mice. The anti-tumor activity of NGR-SSL-PTX following metronomic administration was evaluated in HT1080 tumor-bearing mice in vivo. The targeting activity of the NGR-modified liposomes was demonstrated by in vitro flow cytometry and confocal microscopy as well as in vivo confocal immunofluorescence microscopy and bio-distribution experiments. The results of endothelial cell proliferation and migration and microvessel density (MVD) confirmed the anti-angiogenic activity of NGR-SSL-PTX in vitro and in vivo. The sustained circulation of NGR-SSL-PTX was shown in the pharmacokinetic study. NGR-SSL-PTX is able to improve treatment efficacy producing the most significant anti-tumor activity and anti-angiogenic following metronomic administration.

Analysis of acute radiation-induced esophagitis in non-small-cell lung cancer patients using the Lyman NTCP model.

PURPOSE: To analyze acute esophagitis (AE) in a Chinese population receiving 3D conformal radiotherapy (3DCRT) for non-small cell lung cancer (NSCLC), combined or not with chemotherapy (CT), using the Lyman-Kutcher-Burman (LKB) normal tissue complication probability (NTCP) model. MATERIALS AND METHODS: 157 Chinese patients (pts) presented with NSCLC received 3DCRT: alone (34 pts) or combined with sequential CT (59 pts) (group 1) or with concomitant CT (64 pts) (group 2). Parameters (TD(50), n, and m) of the LKB NTCP model predicting for>grade 2 AE (RTOG grading) were identified using maximum likelihood analysis. Univariate and multivariate analyses using a binary regression logistic model were performed to identify patient, tumor and dosimetric predictors of AE. RESULTS: Grade 2 or 3 AE occurred in 24% and 52% of pts in group 1 and 2, respectively (p<0.001). For the 93 group 1 pts, the fitted LKB model parameters were: m=0.15, n=0.29 and TD(50)=46 Gy. For the 64 group 2 pts, the parameters were: m=0.42, n=0.09 and TD(50)=36 Gy. In multivariate analysis, the only significant predictors of AE were: NTCP (p<0.001) and V(50), as continuous variable (RR=1.03, p=0.03) or being more than a threshold value of 11% (RR=3.6, p=0.009). CONCLUSIONS: A LKB NTCP model has been established to predict AE in a Chinese population, receiving thoracic RT, alone or combined with CT. The parameters of the models appear slightly different than the previous one described in Western countries, with a lower volume effect for Chinese patients.

[Effects of different factors on the expression of thymic stromal lymphopoietin in respiratory syncytial virus-infected human airway epithelial cells].

OBJECTIVE: To investigate the effects of different factors on the expressions of thymic stromal lymphopoietin (TSLP) in respiratory syncytial virus (RSV)-infected human airway epithelial cell line 16HBE cells. METHODS: RSV amplified by infecting Hep-2 cells was identified for its virulence. 16HBE cells were divided into six groups, namely the control group, RSV group, RSV/anti-TLR3 group, RSV/IFN-gamma group, RSV/IL-4 group and RSV/dexamethasone group with corresponding treatments. Real-time RT-PCR was used to examine the expression of TSLP mRNA in the cells 6 h after RSV infection. Western blotting was used to examine TSLP protein expression in the cells 24 h after the infection. RESULTS: The expression of TSLP mRNA in 16HBE cells 6 h after RSV infection increased by 1.63-/+0.08 folds as compared to the expression level in the control cells. The expression of TSLP mRNA was significantly decreased in RSV-infected cells treated with anti-TLR3 antibody (P=0.034) and recombinant human IFN-gamma (P<0.001), but increased with the treatment by recombinant human IL-4 (P=0.025). Dexamethasone significantly inhibited the expression of TSLP mRNA in RSV-infected cells (P<0.001). The production of TSLP protein in 16HBE cells increased by 1.9 folds (P<0.001) 24 h after RSV infection, but underwent no significant changes after treatment with anti-TLR3 antibody (P=0.114). Recombinant human IFN-gamma significantly decreased while IL-4 enhanced the expression of TSLP protein in the infected cells (P=0.020 and 0.014, respectively). Dexamethasone significantly inhibited the increment of TSLP protein expression in RSV-infected cells (P<0.001). CONCLUSIONS: RSV infection can enhance the expressions of TSLP in human airway epithelial cells. IFN-gamma, anti-TLR3 and dexamethasone can inhibit the elevation of TSLP expression induced by RSV infection, but IL-4 synergistically enhances its expression.

[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.

[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.

[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.

[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 effects of recombinant human endostatin on growth and metastasis of lung adenocarcinoma LA795 in mice].

BACKGROUND & OBJECTIVE: Generally, growth and metastasis of tumor are critically dependent on angiogenesis. Endostatin can specifically inhibits tumor angiogenesis. The current study was designed to evaluate the inhibitory effect of recombinant human endostatin (rhES) secreted by pichia, pastoris, GS115 on the growth and metastasis of mice lung adenocarcinoma LA795 in mice T739. METHODS: To select a strain that could highly express recombinant human endostatin, then induce the clone to express rhES by adding methanol. Purification of rhES was performed with heparin affinity chromatography. LA795 tumor cells were inoculated subcutaneously into the dorsa of T739 mice, and the mice were randomized into two groups. The first group was given rhES(20 mg/kg/d), and the second group was given equal volume of PBS, for 14 consecutive days. The volume of tumors were measured. And the tumor metastasis in the lungs of the mice was observed. RESULTS: The selected clone was induced to secrete enough soluble rhES. The purified protein could strongly inhibit growth and metastasis of mice lung adenocarcinoma LA795 in T739 mice (P < 0.001). CONCLUSION: The rhES secreted by pichia, pastoris, GS115 has good biological activities and greatly inhibit growth and metastasis of mice lung adenocarcinoma LA795 in mice T739.

[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.