Proteomic analysis of liver cancer cells treated with suberonylanilide hydroxamic acid

Purpose Suberonylanilide hydroxamic acid (SAHA) is an orally administered histone deacetylase inhibitor (HDACI) that has shown significant antitumor activity in a variety of tumor cells. To evaluate if SAHA has an activity against liver cancer, and with an aim to identify the altered cellular factors upon SAHA treatment, human HepG2 cancer cell line was used as a model, and proteomic approach was utilized to elucidate the molecular mechanisms underlying SAHA’s antitumor activity. Methods Cell growth inhibition was measured by MTT method, and apoptosis was detected by means of flow cytometry analysis and TUNEL assay. Protein expression profiles were analyzed by 2-DE coupled with MALDI-Q-TOF MS/MS analysis. Results A total of 55 differentially expressed proteins were visualized by 2-DE and Coomassie Brilliant Blue (CBB) staining. Of these, 34 proteins were identified via MS/MS analysis. Among the identified proteins, six proteins also displayed significant expression changes at earlier time points upon SAHA treatment, and such alterations were further confirmed by semi-quantitative RT-PCR. Together, at both the mRNA and protein levels, SAHA suppressed the expression of reticulocalbin 1 precursor (RCN1), annexin A3 (ANXA3) and heat shock 27 kDa protein 1 (HSP27), while increasing the expression of aldose reductase (AR), triosephosphate isomerase 1 (TPI) and manganese superoxide dismutase (SOD2). Conclusion SAHA remarkably inhibited proliferation of HepG2 cancer cells, and induced apoptosis in vitro. Using proteomics approaches, a variety of differentially expressed proteins were identified in HepG2 cancer cells before and after treatment with SAHA. This study will enable a better understanding of the molecular mechanisms underlying SAHA-mediated antitumor effects at the protein level.     

Frequent inactivation of RUNX3 by promoter hypermethylation and protein mislocalization in oral squamous cell carcinomas

Purpose  RUNX3 is a functionally important component in transforming growth factor-beta (TGF-β) mediated signaling pathway. Epigenetic silencing expression of RUNX3, as well as aberrant cytoplasmic retention of RUNX3 protein are causely involved in gastric carcinogenesis. Here, we examined the expression of RUNX3 gene and protein in oral squamous cell carcinomas (OSCCs) and analyzed the methylation status of RUNX3 promoter region. Methods  About 10 normal oral mucosa and 30 OSCCs were collected to examine RUNX3 expression by RT-PCR analysis and immunohistochemistry assay using anti-RUNX3 monoclonal antibody R3-6E9. Methylation-specific PCR was carried out on the same specimens to analyze the methylation status of RUNX3 promoter. In addition, the stored paraffin-embedded specimens, including 40 oral leucoplakia (OLK) and 120 OSCCs, were examined by immunohistochemistry assay. Results  RUNX3 gene and protein were underexpressed in OSCCs due to promoter hypermethylation. Protein mislocalization occured frequently. Both downregulation of RUNX3 protein expression (P = 0.001) and protein mislocalization (P = 0.001) were correlated with the differentiation grades in OSCCs. Conclusions  RUNX3 plays an important role in oral carcinogenesis. It may be a useful diagnostic marker and a potential therapeutic target for OSCC. F. Gao and C. H. Huang contributed equally to this article.     

Dissection of aberrant GPCR signaling in tumorigenesis--a systems biology approach

The superfamily of G-protein-coupled receptors (GPCRs) is one of the largest mammalian protein families. It is involved in signal transduction and participates in the regulation of normal physiological function and pathological progression of a range of diseases. Recent studies have demonstrated that many aberrant GPCRs and their ligands are associated with tumorigenesis, angiogenesis and metastasis, which provides promising opportunities for drug discovery for cancer prevention and treatment. It is necessary to search for drug targets such as ligands of unknown GPCRs and better modulators of known GPCRs using high throughput screening approaches. Here, we review recent research advances in the identification of novel GPCRs and their protein interactions.     

Image monitoring for an intraperitoneal bleeding model of pigs using electrical impedance tomography

Current medical imaging techniques are not effective for timely detection of internal hemorrhage when the bleeding is slow and in small quantities. In this study, electrical impedance tomography (EIT) was applied to monitor the intraperitoneal bleeding of an animal model. Five healthy pigs three months old were used. The process of intraperitoneal bleeding was simulated with the injection of anticoagulated blood which was controlled by an electronic syringe pump. The injected rate was no more than 100 ml h(-1) and the total injection volumes ranged from 300 ml to 500 ml. Sixteen electrodes were attached to the abdomen and used for electrical current excitation and surface voltage measurement. Dynamic changes in impedance distribution within the abdomen were calculated by the back-projection algorithm and a series of EIT images were displayed in a unified range. The monitoring was performed with EIT at a rate of one frame per second and continued for at least 4 h. Intraperitoneal blood volume changes could be identified by inspection of consecutive EIT images during the progression of blood injection. 30 ml of blood in the peritoneum could be detected. EIT was shown to be a promising technique for continuous monitoring of intraperitoneal bleeding over periods of time.     

Cys–X–Cys ligand 9 might be an immunological factor in the pathogenesis of oral submucous fibrosis and its concomitant oral lichenoid lesion

Objectives

Oral submucous fibrosis (OSF) is a chronic oral precancerous disease primarily caused by betel quid chewing. Some OSF patients are concomitant with oral lichenoid lesion (OLL), a white-streak lesion with a higher risk for cancerization, in OSF mucosa. Immunological reaction has been considered as one of their common pathogenic mechanisms. Cys–X–Cys ligand 9 (CXCL9) is an important factor to recruit effector neutrophils and lymphocytes in immunological reactions. However, the expression levels of CXCL9 in OSF and OLL remain unclear.

Materials and methods

We investigated the expression levels of CXCL9 in 10 normal buccal mucosa (NBM) samples and 56 OSF concomitant with OLL patients, and evaluated the possible mechanism of CXCL9 on their pathogenesis.

Results

Our results showed NBM demonstrated negative CXCL9 expression. OSF stained positive CXCL9 mainly in the cytoplasm of inflammatory cells and endothelial cells throughout the superficial layer of connective tissue, while its concomitant OLL showed much stronger CXCL9 in all mononuclear cells of subepithelial inflammatory infiltration (p?=?0.0006). There was an upregulated trend of CXCL9 expression from NBM to OSF to OLL. However, no significant association between CXCL9 expression and clinicopathologic parameters of patients was found.

Conclusions

In conclusion, CXCL9 was found for the first time to contribute to the immunological pathogenesis for both OSF and its concomitant OLL, indicating a continuously enhanced intensity of immunoreactivity in their pathogenic process.

Clinical relevance

CXCL9 might be a useful tool to monitor the phase and disease severity of OSF and OLL, and a potential target for further clinical therapy for both lesions.     

接触性导管溶栓治疗急性左下肢深静脉血栓形成的单中心经验

［摘要］ 目的 探讨采用接触性导管溶栓治疗急性左下肢深静脉血栓形成的临床疗效。方法 收集我科2010年06月~2014年01月期间行导管溶栓治疗38例急性左下肢深静脉血栓形成的临床资料,所有患者均采用接触性导管溶栓治疗。比较治疗前后双下肢周径差、溶栓并发症、血栓后综合征等疗效指标。结果 所有患者经溶栓治疗后痊愈或好转后出院,出院前双下肢髌骨下10 cm周径差由3.74±0.94 cm下降为1.85±0.76 cm;双下肢髌骨上10 cm周径差径差由7.31±0.91 cm下降为4.35±1.28 cm,经配对样本t检验,P<0.05,治疗效果显著。平均随访18±9.3个月,随访成功率100%,随访期间10例（26.3%）患者诊断为血栓后综合征。结论 采用导管溶栓治疗急性下肢深静脉血栓形成的疗效确切、血栓后综合征发生率较低,是安全有效的治疗手段。     

Effect of p38 mitogen-activate protein kinase on MUC5AC protein expression of bile duct epithelial cells in hepatolithiasis patients

Primary hepatolithiasis is a common bile duct disease with benign nature but complicated mechanisms. Current studies have revealed its correlation with cytokine release by chronic inflammation, which also increased mucin (MUC) synthesis. This study investigated the role of p38 mitogen-activated protein kinase (MAPK) in regulating cytokine release and mucin synthesis, in an attempt to elucidate the role of p38 signaling molecule in the pathogenesis of hepatolithiasis. In human intrahepatic bile duct endothelial cells (HIBECs), lipoprotein (LPS) was used to induce the high expression of MUC. Small interference RNA (siRNA) was then used to silencing p38 gene expression. Cytokines including interleukin (IL)-1β and tumor necrosis factor (TNF)-α were measured, along with MUC5AC protein and mRNA expression assay. The interference of p38 gene expression inhibited the release of IL-1β and TNF-α in cultured cells. It also depressed both mRNA and protein levels of MUC5A. P38 MAPK signal pathway may be involved in the formation and progression of hepatolithiasis. This study provides potential new strategy for treating hepatolithiasis using p38 MAPK signal pathway as the drug target.     

Redox Regulation of Inflammation: Old Elements,a New Story

Inflammation is an essential immune response characterized by pain, swelling, redness, heat, and impaired function. A controlled acute inflammatory response is necessary to fight off infection and overcome injury. However, if the inflammatory process persists and enters into the chronic state, it can lead to local and systemic deleterious effects counterproductive to healing and instead constitutes a new pathology. Typically, inflamed tissues are associated with an elevated level of reactive species (reactive oxygen species (ROS)/reactive nitrogen species (RNS)). These ROS/RNS are generated during the respiratory burst of immune cells and are important factors in defense against invading pathogens. Additionally, reactive species are now known to trigger oxidative/nitrosative modifications of biomolecules. While most of these modifications lead to irreparable damage, some are subtle and fully reversible. The reversible modifications can initiate signaling cascades known as “redox signaling.” This redox signaling tightly modulates the inflammatory response. Thus, understanding the complex role of ROS/RNS‐induced redox signaling in inflammation will assist in the design of relevant therapeutic intervention strategies for inflammation‐associated diseases. This review will highlight the impact of oxidative stress and redox signaling on inflammation and inflammation‐associated diseases, with a focus on redox modifications of inflammation‐related proteins.     

乳牙外伤致继承恒牙萌出前冠内病损1例

乳牙外伤可累及其继承恒牙胚,导致继承恒牙发育异常、萌出异常,严重时不得不被拔出。该文报告1例乳牙外伤导致继承恒牙发生萌出前牙冠内病损、并伴发阻生、釉质发育不全、牙根发育畸形。