Retraction Note: Antisense oligonucleotides targeting midkine induced apoptosis and increased chemosensitivity in hepatocellular carcinoma cells.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Retraction Note: Antisense oligonucleotides targeting midkine inhibit tumor growth in an in situ human hepatocellular carcinoma model.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

DIP2C expression in breast cancer and its clinical significance.

INTRODUCTION: The aim of this study was to investigate DIP2C expression in different subtypes of breast cancer tissues and cell lines and its correlation with clinicopathologic and histopathological features, in an effort to elucidate the DIP2C expression profile in breast cancer and its clinical significance. METHODS: Hereby, we investigated the DIP2C expression in breast cancer tissues using TMA-IHC method and the DIP2C expression in breast cell lines using quantitative RT-PCR. RESULTS: DIP2C displayed universal expression, being present in all the breast cancer subtypes. There were more cases that staining weakly in breast cancer tissues (n=79/150, 52.7%) than that in fibroadenomas tissues (n=2/18, 11.1%) and normal tissues (n=2/20, 10.0%) (χ2=21.84, P <0.001). Within different intrinsic subtypes of breast cancer assayed by IHC expression profiles, there were less cases of the strongly staining group in basal-like subtype (n=38/86, 44.2%) and HER-2 subtype (n=6/24, 25.0%) than that in luminal A (14/20, 70%) and luminal B (13/20, 65%) subtypes (χ2=11.77, p=0.008). Furthermore, DIP2C expression was positive correlated with ER (χ2=8.90, p=0.003) and PR expression (χ2=10.94, p=0.001), while negative correlated with EGFR expression (χ2=9.27, p=0.002), in accordance with the results of cell lines with different subtypes. Oncomine database also confirmed that, DIP2C was expressed lower in breast cancer tissues, and could indicate prognosis. CONCLUSION: our data revealed DIP2C expression level decreased in breast cancer, especially in basal-like and HER-2 subtypes, and could be a valuable target for diagnosis on specific subtype of breast cancer.

Deregulation of miR-126-3p in basal-like breast cancers stroma and its clinical significance.

INTRODUCTION: The aim of this study was to investigate miR-126-3p expression in stroma and tumor cells of basal-like breast cancer tissues, in an effort to elucidate the potential effect of miR-126-3p on tumor microenvironment and progress of basal-like breast cancer. METHODS: Expression levels of miR-126-3p in 33 paired basal-like breast cancer tissues were assayed by real-time quantitative PCR. Tumor cells and normal epithelial cell were isolated from ten paired basal-like breast cancer tissues and matched adjacent tissues, separately, using laser capture microdissect(LCM)-based PCR method. Further validated in larger sets were assayed by tissue microarrays (TMA)-based ISH method. RESULTS: MiR-126-3p expression level had no significant differences between basal-like breast cancer subtypes and matched adjacent tissues. However, a decreasing trend of miR-126-3p expression can be found in tumor cells of basal-like subtype, compared with matched adjacent tissues, using LCM-based PCR. Using TMA method, miR-126-3p expression level was the lowest in stroma of basal-like breast cancers among four subtypes (χ2=10.55, P=0.01), and was increasing in stroma of breast cancers compared with fibroadenomas. Furthermore, strong miR-126-3p expression in stroma is significantly associated with HER-2 expression (χ2=4.70, P=0.03) and Ki-67 index. (χ2=4.84, P=0.03), which suggested a potential prognostic value of miR-126-3p in stroma of breast cancer. However, miR-126-3p expression in tumor cells derived from different subtypes hadn't significant clinical values in this study. CONCLUSIONS: the miR-126-3p expression level in breast cancer stroma was associated with different intrinsic subtypes and its correlation with hormone receptor and Ki-67 index shed light on the potential clinical prognostic value of miR-126-3p, in the field of specific breast cancer subtypes.

[Imaging Findings of Ovarian Fibrothecoma].

OBJECTIVE: To summarize the computed tomography (CT) and magnetic resonance imaging (MRI) characteristics of ovarian fibro the coma and to compare them with the pathological findings. METHOD: CT and MRI features of 25 patients with pathologically proved ovarian fibrothecoma were retrospectively analyzed. RESULTS: Of these 25 patients,the tumors were single in 23 patients and bilateral in 2 patients. The tumors were round or oval in 16 cases and lobulated in 11 cases. CT plate scanning showed that both the solid masses and the solid components of the cystic and solid masses had slightly lower densities than that of the myometrium, and gradual and mild enhancement could be found in the arterial phase and delay phase after enhanced scan. MRI showed iso-low signal on T1-weighted imaging, slightly low or high signal on T2-weighted imaging of fat suppression sequences, slightly high signal on diffusion weighted imaging and the enhance characteristics as the well as CT after enhanced scan.Pelvic cavity effusion was seen in 3 cases. CONCLUSION: Ovarian fibrothecoma have certain imaging characteristics,which are helpful to improve the diagnosis and differential diagnosis of this disease and lower the misdiagnosis rate before operation.

Inhibitory effect of midkine-binding peptide on tumor proliferation and migration.

BACKGROUND: To investigate the inhibitory effect of midkine-binding peptides on human umbilical vein endothelial cells (HUVECs) proliferation and angiogenesis of xenograft tumor. METHODS: The midkine-binding peptides were panned by Ph.D.-7(™) Phage Display Peptide Library Kit, and the specific binding activities of positive clones to target protein were examined by phage ELISA. The effect of midkine-binding peptides on proliferation of HUVECs was confirmed by MTT test. The xenograft tumor model was formed in BALB/c mice with the murine hepatocarcinoma cells H22 (H22). Microvessel density (MVD) was analyzed by immunohistochemistry of factor VIII staining. RESULTS: Midkine-binding peptides have the inhibitory effects on tumor angiogenesis, a proliferation assay using human umbilical vein endothelial cells (HUVECs) indicated that particular midkine-binding peptides significantly inhibited the proliferation of the HUVECs. Midkine-binding peptides were also observed to efficiently suppress angiogenesis induced by murine hepatocarcinoma H22 cells in BALB/c nude mice. CONCLUSION: The midkine-binding peptides can inhibit solid tumor growth by retarding the formation of new blood vessels. The results indicate that midkine-binding peptides may represent potent anti-angiogenesis agents in vivo.

Upregulation of miR-1280 expression in non-small cell lung cancer tissues.

BACKGROUND: Non-small cell lung cancer (NSCLC) is a prolific and high-mortality disease with few effective treatments. Although the detection and surgical techniques for NSCLC continue to advance, the survival rate for the patients with NSCLC remains poor. Enhanced predictive biomarkers such as microRNAs (miRNAs) are needed at the time of diagnosis to better tailor therapies for patients. This study focused on the expression of miR-1280 in NSCLC tissues and distal normal tissues in order to explore the association between miR-1280 expression and NSCLC. METHODS: A total of 72 newly diagnosed primary NSCLC patients were enrolled in this study. Quantitative real-time polymerase chain reaction (PCR) was performed to identify the expression level of miR-1280 in the NSCLC tissues and distal normal tissues of these patients. RESULTS: The miR-1280 expression was significantly higher in the NSCLC tissues (0.084 ± 0.099) than distal normal tissues (0.014 ± 0.015, P = 0.009). In 54 patients (75%), the miR-1280 expression in the NSCLC tissues was upregulated (2-ΔΔct > 2), and no case showed a downregulation of miR-1280 expression. CONCLUSIONS: The expression level of miR-1280 could be regarded as a biomarker for NSCLC.

[IgG4-associated parotitis:case report and review of the literature].

A case of immunoglobulin G4 (IgG4)-associated parotitis was reported and related literatures were reviewed,in order to improve the recognization of this systemic diseases and reduce the misdiagnosis and mistreatment in clinical practice for the stomatologists.

[Correlation between androgen receptor expression and hepatitis B virus X protein and its clinical significance in hepatocellular carcinoma].

OBJECTIVE: To investigate the expression of androgen receptor (AR) and hepatitis B virus X protein (HBx) in hepatocellular carcinoma (HCC), and analyze the relationship between AR and HBx expressions. METHODS: Tumor tissues and peritumoral tissues of 83 HBV-associated HCC cases were investigated in this study. Fourteen cases of HBV-negative HCC and 13 cases of hemangioma peritumoral tissues were considered as control. AR and HBx mRNA levels were determined by quantitative fluorescence real-time RT-PCR and their protein levels were assayed by Western blot. The expression of AR and HBx proteins in tissues were examined with EnVision immunohistochemical staining. The methylation status of AR promoter was determined using methylation-specific PCR (MSP). RESULTS: Both expression levels of AR mRNA and protein of the peritumoral tissues were significantly higher (0.17) than that of tumor tissues (0.09) in HBV-associated HCC (P < 0.01), but such a difference was not found in HBV-negative HCC (0.06 vs. 0.07, P > 0.05). The level of AR expression in peritumoral tissues was associated with tumor differentiation in HBV-associated HCC. AR mRNA and protein levels of peritumoral tissues in HBV-associated HCC were significantly higher than that in HBV-negative HCC and hemangioma (all P < 0.05). In the tumor tissues, HBV-associated HCC had significantly higher AR expression than HBV-negative HCC at mRNA level (P < 0.05), but not at protein level. Spearman rank correlation analysis showed that the AR mRNA or AR protein levels were positively correlated with HBx in both tumor and peritumoral tissues in HBV-associated HCC, but the expressions of AR and HBx were not associated with AR promoter methylation status. The relative expression levels of AR mRNA and protein in the HBV-associated peritumoral tissues were negatively correlated with tumor differentiation (r = -0.213, P < 0.05; r = -0.313, P < 0.05), the higher the AR expression, the poorer differentiation. But this correlation of AR mRNA and protein was not shown in the hepatocellular carcinoma tissues. CONCLUSIONS: HBx may enhance AR expression in HBV-associated HCC, but AR promoter demethylation maybe not been involved in its main mechanism. An increased AR expression is probably an early event during the development and progression of HBV-associated HCC, and AR expression in the peritumoral tissue is correlated with HBV-associated HCC differentiation. AR may play different roles in HBV-associated HCC and HBV-negative HCC.

Progranulin expression in breast cancer with different intrinsic subtypes.

Progranulin is a newly discovered 88-kDa glycoprotein originally purified from the highly tumorigenic mouse teratoma-derived cell line PC. We found that high progranulin expression was associated with higher breast carcinoma angiogenesis, reflected by increased vascular endothelial growth factor expression and higher microvessel density. However, no immunohistochemical evidence currently exists to correlate progranulin expression with clinicopathological features in different intrinsic subtypes of breast carcinoma biopsies. The aim of this study was to investigate the progranulin expression profiles in the intrinsic subtypes of breast carcinomas and their relevance to histopathological and clinicopathological features. Tissue blocks containing 264 cases of breast carcinomas from 2006 to 2009 were classified as different intrinsic subtypes. Tissues of four intrinsic subtypes were immunostained for progranulin, vascular endothelial growth factor and CD105. Their relevance to histopathological and clinicopathological features was also analyzed. Twenty tissue samples from breast fibroadenomas were included in this study. Progranulin expression showed no significant differences in different intrinsic subtypes, although an increasing tendency could be found in the triple-negative breast cancer (TNBC) subgroup (χ(2)=5.00, df=3, p=0.17). However, differences were significant when pathologically node metastasis-positive (pN(+)) TNBC were excluded (χ(2)=17.84, df=3, p<0.01). Some clinicopathological parameters, including CK5/6 (χ(2)=0.08, df=3, p=0.78), E-cadherin (χ(2)=0.71, df=3, p=0.40) and P53 (χ(2)=0.05, df=3, p=0.83), displayed no correlation with activity of progranulin in pathologically node metastasis-negative (pN(-)) TNBC. It was noted that the EGFR expression level of the pN(-) TNBC subtype was significantly higher in cases with strong progranulin expression than in cases with weak progranulin expression (χ(2)=11.26, df=1, p<0.01). A significantly higher expression level of progranulin in pN(-) TNBC suggests that progranulin is a promising new target for pN(-) TNBC treatment. Strong expression of progranulin correlates with positive EGFR expression in the pN(-) TNBC subtype. The close relationship between EGFR and progranulin/VEGF/CD105 expression may partly play a role in high angiogenesis levels in the pN(-) TNBC subtype.

Clinicopathologic and prognostic implications of progranulin in breast carcinoma.

BACKGROUND: Progranulin is a newly discovered 88-kDa glycoprotein originally purified from the highly tumorigenic mouse teratoma-derived cell line PC. Its expression is closely correlated with the development and metastasis of several cancers. However, no immunohistochemical evidence currently exists to correlate progranulin expression with clinicopathologic features in breast carcinoma biopsies, and the role of progranulin as a new marker of metastatic risk and prognosis in breast cancer has not yet been studied. The aim of this study was to investigate the clinicopathologic and prognostic implications of progranulin expression in breast carcinoma and its correlation with tumor angiogenesis. METHODS: Progranulin expression was determined immunohistochemically in 183 surgical specimens from patients with breast cancer and 20 tissue samples from breast fibroadenomas. The tumor angiogenesis-related biomarker, vascular endothelial growth factor was assayed and microvessel density was assessed by counting vascular endothelial cells in tumor tissues labeled with endoglin antibody. The relationship between progranulin expression and the clinicopathologic data were analyzed. RESULTS: Progranulin proteins were overexpressed in breast cancer. The level of progranulin expression was significantly correlated with tumor size (P = 0.004), lymph node metastasis (P < 0.001) and TNM staging (P < 0.001). High progranulin expression was associated with higher tumor angiogenesis, reflected by increased vascular endothelial growth factor expression (P < 0.001) and higher microvessel density (P = 0.002). CONCLUSION: Progranulin may be a valuable marker for assessing the metastasis and prognosis of breast cancer, and could provide the basis for new combination regimens with antiangiogenic activity.

[Effects of combined enteral nutrition support on hemorrheologic parameters and the level of inflammatory factors in rabbits with severe acute pancreatitis].

OBJECTIVE: To explore the effects of combined enteral nutrition (CEN) on the hemorheologic parameters and the changing levels of inflammatory factors in an animal model of severe acute pancreatitis (SAP). METHODS: The experimental animals were divided randomly into 3 groups, i.e. early enteral nutrition (EEN) group, CEN group and parenteral nutrition(PN)group (n = 20 each). Enteral nutrition was administered to the EEN and CEN group animals at 24 h and 72 h post-modeling respectively. The PN group animals were supported by parenteral nutrition all time. Hemorrheologic indices of all experimental animals were examined on Days 1, 3 and 7 post-modeling. And the inflammatory factors were examined on Days 1 and 7. RESULTS: Compared with the EEN and PN groups, some hemorrheologic indices of the CEN group decreased significantly (P < 0.05) on Day 7 post-modeling. They included blood sedimentation, hematocrit (HCT), whole blood high-cut reduction viscosity and whole blood low-cut reduction viscosity. As compared within the CEN group, each hemorrheologic index was lower on Day 7 than that on Day 1 (P < 0.05). Except for whole blood high-cut reduction viscosity and erythrocyte aggregation index in the EEN group after a 7-day nutrition support, there was no significant change for all hemorrheologic indices in the PN group. As to the level of inflammatory factors, the values of interleukin 8 (IL-8) and tumor necrosis factor-α (TNF-α) in the CEN group were lower than those in the PN group on Day 7 post-modeling (P < 0.05). The values of IL-8 and IL-6 in the CEN group were lower than those in the EEN group on the same day (P < 0.05). As compared within the CEN group, the values of IL-6 and TNF-α were lower on Day 7 than those on Day 1 post-modeling (P < 0.05). CONCLUSION: The modulatory mechanism of EN over SAP should be achieved by correcting hemorrheologic index change and lowering the level of inflammatory factors. A proper timing of EN is probably the most optimal nutrition support mode of SAP therapy.

Nuclear factor-κB and apoptosis in patients with intracerebral hemorrhage.

Studies using rat models have indicated that neuronal apoptosis is involved in the pathogenesis of intracerebral hemorrhage (ICH); however, the mechanism by which apoptosis occurs is unclear. In the present study, we aimed to quantify the number of nuclear factor-κB (NF-κB)-positive cells and apoptotic cells in specimens of middle temporal gyrus taken from 46 human subjects with hypertensive ICH. We also investigated the roles that intercellular adhesion molecule-1 (ICAM-1) and interleukin (IL)-1ß play in apoptosis following ICH. At about 24 hours after ICH, some neurons exhibited nuclear swelling and incomplete cellular structures were visible. The mean percentage of apoptotic cells was 39.28 ± 21.83% at 49-72 hours after ICH. NF-κB immunoreactivity varied with time after ICH: the number of immunostained neurons increased during the 2-6 hours after ICH, and reached a maximum at 7-48 hours. The number of IL-1ß-immunostained neurons reached a maximum at 2-6 hours after ICH. The number of ICAM-1-immunostained neurons increased during the 48 hours after ICH and reached a maximum at 49-72 hours. These observations indicate that apoptosis has a major role in pathological cell death after ICH and that activation of NF-κB is positively related to the progress of apoptosis. Additionally, activation of ICAM-1 and IL-1ß seem to be involved in apoptosis after ICH.

Expression of midkine and endoglin in breast carcinomas with different immunohistochemical profiles.

The aim of this study was to investigate the midkine and endoglin expression in breast carcinomas with five different immunohistochemical profiles and their relevance to histopathologic and clinicopathologic features. We analyzed 161 archival tissues immunohistologically. The level of midkine expression in breast cancer significantly correlated with lymph node metastasis (p = 0.001) and TNM staging (p = 0.003). High microvessel density (MVD) was associated with higher midkine reactivity group (p = 0.036). Although the basal-like subtype had higher midkine expression level and MVD, no significant difference with the other breast cancer subtypes was found. In conclusion, midkine was a promising target for tumor prognosis in clinical diagnosis and treatment. This study found no significant differences in tumor angiogenesis in different molecular subtypes of breast cancer.

[Expression characteristics of nuclear factor kappa B in hepatocellular carcinoma tissues].

OBJECTIVE: To investigate the expression characteristics of nuclear factor kappa B (NF-kB) in hepatocellular carcinoma (HCC) tissues and its correlation with tumor necrosis factor alpha (TNF alpha) and clinical pathological features. METHODS: Thirty liver specimens from HCC patients were collected by self-control method. The localization and expression of NF-kappaB in HCC and their surrounding tissues were detected by immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively. And the levels of TNF alpha in these tissues were analyzed by ELISA. RESULTS: The expressed NF-kappaB was localized in nucleus and cytoplasm in HCC, whereas only in cytoplasm in the surrounding tissues. The expression level and density of NF-kappaB in HCC tissues were obviously higher than those in the surrounding tissues (P < 0.01), which was positively correlated with increased TNF alpha in HCC tissues (r = 0.964, P < 0.01). No positive correlation was found between NF-kappaB expression and histological differentiation grade, number of tumor, size of tumor, and HBsAg positive (P > 0.05). CONCLUSION: The expression and localization of NF-kappaB in HCC tissues are obviously different from those in the surrounding normal liver tissues, and the level of nucleoprotein NF-kappaB in HCC tissues is correlated with expressed TNF alpha, suggesting that TNF alpha can activate NF-kB, the activated NF-kB then translocates to the nucleus and plays important role in the carcinogenesis of HCC.

In vitro and in vivo suppression of hepatocellular carcinoma growth by midkine-antisense oligonucleotide-loaded nanoparticles.

AIM: To synthesize antisense oligonucleotides (ASODNs) of midkine (MK), package the ASODNs with nanoparticles, and to inhibit hepatocellular carcinoma (HCC) growth using these nanoparticles. METHODS: HepG2 cell proliferation was analyzed in vitro using the 3-(4,5-dimethythiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt assay. The in vivo activity of nanoparticles delivering the MK-ASODNs was analyzed by histopathological and immunohistochemical staining and quantitative real time polymerase chain reaction (PCR). RESULTS: The in vitro proliferation of HepG2 cells was significantly inhibited by the nanoparticles packaged with MK-ASODNs (NANO-ASODNs). Furthermore, the NANO-ASODNs significantly inhibited the growth of HCC in the mouse model. CONCLUSION: NANO-ASODNs can significantly suppress the growth of HCC in vitro and in vivo.

[An animal experiment of immediate construction of mandibular defect].

PURPOSE: To observe the effect of immediate construction of mandibular defect with or without teeth missing by bone composite and absorbable Bio-gide membrane. METHODS: 40 New Zealand rabbits were randomly divided into 3 groups. For the mandibular defect, group A was filled with nothing, group B was filled with bone composite and Bio-gide membrane, group C was filled with bone composite and Bio-gide membrane as well as autogenous tooth implantation. The rabbits were sacrificed 4,8,12 weeks after surgery for evaluation of gross morphology, X-ray and histopathology. RESULTS: After 12 weeks, the defect in group A was still there but narrower, the defect in group B was filled with new bone, the density of which was same to the bone X-ray, and lamellar bone were under microscope, the implanted teeth in group C had been fixed tightly in 17 animals, no transparent areas in 13 animals from X-ray, but absorbable areas in 13 animals under microscope were noted. CONCLUSIONS: Immediate construction of mandibular defect with bone composite and Bio-guide membrane is feasible and effective,autogeneous tooth implantation on the construction area is also stable.

Enhanced therapeutic effects of combined chemotherapeutic drugs and midkine antisense oligonucleotides for hepatocellular carcinoma.

AIM: To evaluate the effect of combined antisense oligonucleotides targeting midkine (MK-AS) and chemotherapeutic drugs [cisplatin(DDP), 5-fluorouracil (5-FU) and adriamycin (ADM)] on inhibition of HepG2 cell proliferation, and to analyze the efficacy of MK-AS used in combined ADM in in situ human hepatocellular carcinoma (HCC) model. METHODS: HepG2 cells were treated with MK-AS and/or chemotherapeutic drugs mediated by Lipofectin, and cell growth activity was determined by MTS assay. An in situ HCC model was used in this experiment. MK-AS, ADM and MK-AS + ADM were given intravenously for 20 d, respectively. The animal body weight and their tumor weight were measured to assess the effect of the combined therapy in vivo. RESULTS: Combined treatment with MK-AS reduced the IC50 of DDP, 5-FU and ADM in HepG2 cells. MK-AS significantly increased the inhibition rate of DDP, 5-FU and ADM. Additionally, synergism (Q 1.15) occurred at a lower concentration of ADM, 5-FU and DDP with combined MK-AS. Combined treatment with MK-AS and ADM resulted in the more growth inhibition on in situ human HCC model compared with treatment with chemotherapeutic drugs alone. CONCLUSION: MK-AS increases the chemosensitivity in HepG2 cells and in situ human HCC model, and the combination of MK-AS and ADM has a much better in vitro and in vivo synergism.

Antisense oligonucleotide targeting midkine suppresses in vivo angiogenesis.

AIM: To evaluate the effect of antisense oligonucleotide targeting midkine (MK-AS) on angiogenesis in chick chorioallantoic membrane (CAM) and in situ human hepatocellular carcinoma (HCC). METHODS: An in situ human hepatocellular carcinoma (HCC) model and CAM assay were used in this experiment. The effect of MK-AS on angiogenesis was evaluated by cell proliferation assay and hematoxylin-eosin (HE) staining. RESULTS: MK-AS significantly inhibited human umbilical vein endothelial cells (HUVEC) and in situ human HCC growth. At the same time, MK-AS suppressed the angiogenesis both in human hepatocellular carcinoma cell line (HEPG2)-induced CAM and in situ human HCC tissues. CONCLUSION: MK-AS is an effective antiangiogenesis agent in vivo.

[Extraarticular diffuse pigmented villonodular synovitis of the temporomandibular joint: report of one case and review of the literature].

Diffuse pigmented villonodular synovitis (PVNS) in the temporomandibular joint (TMJ) is distinctly rare. This article reported a case of extra-articular type of PVNS of the TMJ, and reviewed the previously reported cases in the literature. It is concluded that PVNS can be diagnosed by characteristic MRI finding, wide local excision including a total synovectomy and postoperative radiotherapy should be adopted.