Breast cancer is the fifth most common cause of cancer deaths in the world, which often spreads first to the axillary lymph nodes (ALN) from the primary tumor. ALN helps clinician stage breast cancer. In addition, it is one of the key prognostic factors for patients with invasive breast cancer. The sentinel lymph node (SLN) is defined as the first regional lymph node to receive lymphatic fluid from a malignant tumor. As a result, it seems possible to assess the complete nodal status with sentinel lymph node biopsy (SLNB), which is attractive and reliable approach for identifying lymph node metastasis. Ductal carcinoma in situ (DCIS) is the most common type of non-invasive breast cancer. However, the role of SLNB in DCIS is controversial because DCIS does not cause invasion and metastasis theoretically. In this review, clinical applications of SLNB in DCIS will be discussed. The potential benefit of accurately upstaging patients with DCIS and the minimal invasiveness of SLNB justify use of SLNB in selected high-risk DCIS patients. At least DSIS with microinvasion, have DCIS of sufficient extent on mammography or MRI, or indicated invasive or microinvasive focus by final histological examination, are recommended for SLNB. Moreover, large randomized trials to evaluate the usefulness of SLNB in DCIS patients after long-term follow-up on local control and survival are required for further evaluation.
Mesenchymal stem sells (MSCs) have received much attention in the field of bone tissue engineering due to their biological capability to differentiate into osteogenic lineage cells. Hypoxia-inducible factor 1alpha (HIF-1α) plays an important role in the MSC-related bone regeneration during hypoxia, while core binding factor alpha 1 (Cbfα1) is a transcription regulator that is involved in the chondrocyte differentiation and ossification. In the present study, we investigated the effects of hypoxia on biological capability of MSCs. MSCs were isolated from adult rabbit bone marrow, and were cultured in vitro under normoxia (air with 5% CO2) or hypoxia (5% CO2 and 95% N2). The proliferation of MSCs, alkaline phosphatase (ALP) activity, and production of collagens type I and type III (Col I/III) were examined. The expression levels of HIF-1α and Cbfα1 were measured by real-time PCR and western blot analyses. We found that hypoxia significantly induced the proliferation of MSCs and increased ALP activity and the production of Col I/III. Moreover, hypoxia increased the expression of Cbfα1 mRNA after 12 h, whereas the expression of HIF-1α mRNA was increased after 1 h of hypoxia. Knockdown of HIF-1α expression with a small interfering RNA significantly increased the expression levels of Cbfα1 protein either under the normoxia or hypoxia condition. Our results indicate that hypoxia enhances MSCs to differentiate into osteogenic lineage cells and suggest that Cbfα1 may be negatively regulated by HIF-1α.
Acute-on-chronic hepatitis B liver failure (ACHBLF) refers to liver failure occurring in patients with chronic hepatitis B (CHB) related liver diseases. Interferon-γ (IFN-γ) plays an important role in the exacerbation of liver function. However, the exact mechanism, by which IFN-γ mediates ACHBLF, is not fully understood. Forty patients with ACHBLF, fifteen patients with CHB and ten healthy controls were included in this present study. ELISA was performed to measure the level of serum IFN-γ. The methylation status of IFN-γ promoter in peripheral blood mononuclear cells (PBMCs) was determined using methylation-specific PCR. Model for End-stage Liver Disease (MELD) scoring was performed for evaluating the severity of liver failure. The serum level of IFN-γ in patients with ACHBLF or CHB was significantly lower than that in healthy controls, while the serum IFN-γ level in ACHBLF patients was significantly higher than that in CHB patients. In ACHBLF patients, the level of IFN-γ was positively correlated with total bilirubin and MELD score, but negatively correlated with prothrombin time activity. These results suggest the involvement of IFN-γ in the pathogenesis of ACHBLF. Importantly, the degree of methylation of the IFN-γ gene promoter in ACHBLF patients (60%, 24/40) was significantly lower than that in CHB patients (93%, 14/15), but was higher than that in the control group (20%, 2/10). Furthermore, in ACHBLF patients, the serum IFN-γ level was significantly higher in unmethylation group than that in methylation group. In conclusion, enhanced demethylation of IFN-γ gene promoter in PBMCs may be associated with the onset of ACHBLF.
The incidence of esophageal adenocarcinoma has been rising in many countries, while esophageal squamous cell carcinoma has remained stable or even declined in the same populations over the identical periods. These differences in trends indicate that these cancer subtypes may have a different etiology, which may be caused by lifestyle factors such as alcohol consumption and cigarette smoking. Therefore, a matched case-control study to clarify the risk factors of alcohol and tobacco intake on the development of esophageal adenocarcinoma was collected in Hebei Province of China. The life expectancy of the study area was around 70 years old. In the present study, 98 patients younger than 65 years who were diagnosed with esophageal adenocarcinoma and had initial surgeries (cases) were matched with 294 healthy adults (controls) at a ratio of 1:3 according to sex and age. We found the proportions of drinkers and smokers among cases were 48.0% and 60.2%, respectively, versus 21.2% and 43.5% among controls. Univariate conditional logistic regression analyses revealed that the odds ratios (ORs) showed a nearly monotonic increase for the duration of alcohol consumption and duration of tobacco smoking. Multivariate conditional logistic regression analysis indicated that only alcohol consumption was a significant risk factor for esophageal adenocarcinoma. Additional analysis of the combination of amount and duration of alcohol consumption indicated that heavy drinkers (> 30 ml/day) had significantly higher ORs, irrespective of the duration of alcohol consumption. In conclusion, heavy alcohol consumption increases the risk for esophageal adenocarcinoma independent of the duration of such consumption.
Osteopontin (OPN) is overexpressed in a variety of cancers including hepatocellular carcinoma (HCC), and is likely involved in the process of vasculogenic mimicry (VM) in some tumor cells. In this study, we explored whether OPN plays a role for VM in HCC. Metastatic MHCC97-H human HCC cells and non-metastatic Hep3B human HCC cells were compared for their abilities to establish VM. Three dimensional-culture assays showed that MHCC97-H cells but not Hep3B cells were able to form the chord-like structure that represents VM. Real-time RT-PCR arrays were used to detect gene expression profiles of the two HCC cell lines in three-dimensional culture. PCR array analyses revealed the increased expression of OPN in MHCC97-H cells forming VM compared with Hep3B cells. Small interfering RNA was employed to investigate whether OPN knockdown could influence VM, and the expression of matrix metalloproteinase (MMP)-2, MMP-9 and urokinase-type plasminogen activator (uPA) in MHCC97-H cells. OPN knockdown resulted in a significant decrease in the ability of MHCC97-H cells to form VM, which was accompanied by the down-regulation of MMP-2 and uPA expression. Furthermore, human HCC tissue samples were studied by immunohistochemistry to analyze the correlations between VM and the expression of OPN, MMP-2 and uPA. There existed significant positive correlations between VM and the expression of OPN, MMP-2 and uPA in HCC tissue samples. In conclusion, OPN is required for VM in HCC cells, and its action may be associated with activation of MMP-2 and uPA. OPN-targeted therapeutics may be useful for patients with advanced HCC.
The poor prognosis of liver cancer demands the development of new diagnostic markers and therapeutic strategies. Cancer/testis (CT) antigens are expressed in the testis and cancerous tissues, but not in adult somatic cells. Given their tumor-specific expression, CT antigens are potential molecular markers for tumor diagnosis and targets for cancer immunotherapy. To identify novel CT antigens for liver cancer, we examined mRNA expression of hitherto unknown CT antigen candidates, tudordomain-containing protein (TDRD) 1, 4 and 5 in three types of liver cancer; hepatocellular carcinoma (HCC, n = 28), cholangiocarcinoma (CC, n = 5) and combined HCC-CC (n = 8), with matched non-tumorous liver tissues. The TDRD1, 4 and 5 are known as being specifically expressed in the testis. TDRD1 and 5 are essential for male germ cell development. On RT-PCR analysis, TDRD1 mRNA was expressed in both HCCs and non-tumorous liver tissues, and TDRD5 mRNA was expressed in normal colonic and gastric mucosal tissues. Thus, TDRD1 and TDRD5 are not candidates for CT antigens. TDRD4 mRNA was expressed in the testis but not in other normal tissues, including colonic mucosa, gastric mucosa, and liver tissues. TDRD4 mRNA was expressed in 7 of the 41 liver cancers: 4 HCCs, 1 CC and 2 combined HCC-CCs. The TDRD4 mRNA expression was not significantly associated with patient age, tumor size, pathologic stages, hepatitis B virus infection, or CD133 expression. In conclusion, TDRD4 mRNA is expressed in a subset of liver cancers, and TDRD4 is a candidate CT antigen for liver cancer.
Normal endometrial growth is essential for embryonic implantation and maintenance of pregnancy. The uterine endometrium contains stem cells that are involved in tissue regeneration. Side population cells (SP cells) are an emerging cell population that may be responsible for the regeneration process of uterine endometrium. In this study, we investigated the changes in the distribution of SP cells using a mouse model of uterine endometrial injury that was induced by peritoneal injection of lipopolysaccharide (LPS). The uterine horns were collected 0, 6, 12, and 18 hours after LPS injection. ATP-binding cassette and sub-family G member 2 (Abcg2) is highly expressed on the cellular membrane of some stem and progenitor cells, and was used as a marker for SP cells. Immunohistochemistry demonstrated that Abcg2-positive cells were increased around the uterine endometrial glands from 6 to 12 h after LPS injection. The percentage of Abcg2-positive cells was calculated using flow cytometry. The percentage of stromal SP cells was significantly higher at 6 h after LPS injection, compared with the value before the injection (3.01 ± 0.41% vs. 1.63 ± 0.31%, P < 0.05). To evaluate the influence of ovarian hormones, we implanted pellets containing 17β-estradiol (0.1 mg), progesterone (10 mg), or a combination of 17β-estradiol and progesterone in the bilaterally ovariectomized mice. Ovariectomy abolished the increase in SP cells, which was restored by estradiol, but not by progesterone or the combination treatment. In conclusion, estrogen is required for the increase of SP cells, thereby leading to the regeneration of the uterine endometrium.
Human parvovirus B19 (HPV-B19), a small and non-enveloped DNA virus, causes erythema infectiosum (EI) in children. In adults, however, it is known to cause a variety of symptoms. A 39-year-old woman visited our hospital because of low-grade fever, diarrhea, bilateral leg edema, and numbness in the right arm, one and a half months after her daughter developed EI. We diagnosed her as HPV-B19 infection after her daughter's history and positive test for serum HPV-B19 IgM antibody, together with the continued observations. Two weeks later, she developed dizziness and left hearing difficulty. However, we did not give her any medication. HPV-B19 IgM antibody value (2.4) measured after one month of the onset was decreased to 1.7, 1.1, and 0.9 after two, three, and five months of the onset, respectively. Thus, it took 5 months for the IgM antibody value to become negative. Her symptoms gradually improved along with the decrease in HPV-B19 antibody without any medication. Hearing difficulty and dizziness are not categorized as manifestations of HPV-B19 infection, because these symptoms are very rare. The present report indicates that the symptoms related to inner ear dysfunctions should be added to those associated with adult HPV-B19 infection. In conclusion, we should consider HPV-B19 infection when we evaluate patients with causeless hearing difficulty and dizziness.
Methicillin-resistant Staphylococcus aureus (MRSA) has become a leading cause of infections in both the community and in hospitals. MRSA bacteremia is a serious infection with a very high mortality rate. The aim of this study was to assess the clinical features of MRSA bacteremia and to evaluate predictors of mortality in patients with this infection. The medical records of 83 patients with MRSA bacteremia, who had been admitted to Nagasaki University Hospital between January 2003 and December 2007, were retrospectively reviewed. Underlying disease, presumed source, MRSA sensitivity, Staphylococcal cassette chromosome mec (SCCmec) types, virulence genes and prognosis were evaluated. Of the 83 patients (44 men and 39 women; mean age: 63.7 years) with MRSA bacteremia, 30 (36.1%) had malignancy and 25 (30.1%) had been treated with immunosuppressive drugs. Fifteen patients (18.1%) were intravascular catheter related. SCCmec typeII accounted for 80% of SCCmec types of MRSA isolates. The mortality rate was 39.8% (33/83), which is similar to that of previous reports. The ratio of males to females, the mean age or the body temperature did not differ between survivors and nonsurvivors. Independent predictors associated with mortality in the multivariate analyses are pneumonia (P = 0.016), treatment with VCM (P = 0.039), and transplantation (P = 0.021). We suggest that poor prognosis achieved with VCM is in part due to its low blood concentration and poor tissue penetration. VCM should not be selected when presumed source of MRSA bacteremia is pneumonia.
Sepsis is the major cause of death in intensive care units, despite enormous efforts in the development of antimicrobial therapies. Sepsis is mediated by early [e.g., tumor necrosis factor (TNF)-α and interleukin (IL)-1β] and late [e.g., high-mobility group box 1 protein (HMGB1)] proinflammatory cytokines. HMGB1, which is secreted into extracellular milieu by activated macrophages or passively released by destroyed macrophages, stimulates intensive inflammatory responses. D-Ala2-D-Leu5-enkephalin (DADLE), a synthetic δ-opioid receptor agonist, has been shown to protect rats from sepsis. Here we elucidated the mechanism for protective effect of DADLE against sepsis. Sepsis was established in Sprague-Dawley rats by means of cecal ligation and puncture (CLP). In this model, the serum levels of TNF-α and IL-1β were increased after 2-3 h, while those of HMGB1 were increased after 18 h. Administration of DADLE (5 mg/kg) concurrently with CLP improved survival, which was associated with the decreases in the serum levels of TNF-α, IL-1β and HMGB1. Importantly, DADLE administrated 4 h after CLP showed comparable protective effect as the concurrent administration, with decreased serum HMGB1 levels. Moreover, peritoneal macrophages isolated from rats were challenged with lipopolysaccharide (LPS). Concurrent or delayed DADLE administration at 10−6 M suppressed the LPS-induced cell death. DADLE also suppressed the release of HMGB1 from macrophages that was induced by LPS, TNF-α or interferon-γ. In conclusion, DADLE protects rats from sepsis probably by decreasing the serum level of HMGB1. We propose DADLE as a candidate for septic shock therapy, even if it is administered after the onset of sepsis.