TMEM65 promotes gastric tumorigenesis by targeting YWHAZ to activate PI3K-Akt-mTOR pathway and is a therapeutic target.

Copy number alterations are crucial for the development of gastric cancer (GC). Here, we identified Transmembrane Protein 65 (TMEM65) amplification by genomic hybridization microarray to profile copy-number variations in GC. TMEM65 mRNA level was significantly up-regulated in GC compared to adjacent normal tissues, and was positively associated with TMEM65 amplification. High TMEM65 expression or DNA copy number predicts poor prognosis (P < 0.05) in GC. Furtherly, GC patients with TMEM65 amplification (n = 129) or overexpression (n = 78) significantly associated with shortened survival. Ectopic expression of TMEM65 significantly promoted cell proliferation, cell cycle progression and cell migration/invasion ability, but inhibited apoptosis (all P < 0.05). Conversely, silencing of TMEM65 in GC cells showed opposite abilities on cell function in vitro and suppressed tumor growth and lung metastasis in vivo (all P < 0.01). Moreover, TMEM65 depletion by VNP-encapsulated TMEM65-siRNA significantly suppressed tumor growth in subcutaneous xenograft model. Mechanistically, TMEM65 exerted oncogenic effects through activating PI3K-Akt-mTOR signaling pathway, as evidenced of increased expression of key regulators (p-Akt, p-GSK-3ß, p-mTOR) by Western blot. YWHAZ (Tyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase) was identified as a direct downstream effector of TMEM65. Direct binding of TMEM65 with YWHAZ in the cytoplasm inhibited ubiquitin-mediated degradation of YWHAZ. Moreover, oncogenic effect of TMEM65 was partly dependent on YWHAZ. In conclusion, TMEM65 promotes gastric tumorigenesis by activating PI3K-Akt-mTOR signaling via cooperating with YWHAZ. TMEM65 overexpression may serve as an independent new biomarker and is a therapeutic target in GC.

Single-Molecule Observation of Selenoenzyme Intermediates in a Semisynthetic Seleno-α-Hemolysin Nanoreactor.

The development of monitoring methods to capture short-lived intermediates is crucial for kinetic mechanism validation of enzymatic reaction steps. In this work, a semisynthetic selenoenzyme nanoreactor was constructed by introducing the unnatural amino acid (Sec) into the lumen of the α-hemolysin (αHL) nanopore. This nanoreactor not only created a highly confined space to trap the enzyme-substrate complex for a highly efficient antioxidant activity but also provided a single channel to characterize a series of selenoenzyme intermediates in the whole catalytic cycle through electrochemical analysis. In particular, the unstable intermediate of SeOH can be clearly detected by the characteristic blocking current. The duration time corresponding to the lifetime of each intermediate that stayed within the nanopore was also determined. This label-free approach showed a high detection sensitivity and temporal-spatial resolution to scrutinize a continuous enzymatic process, which would facilitate uncovering the mysteries of selenoenzyme catalysis at the single-molecule level.

Supramolecularly regulated artificial transmembrane signal transduction for 'ON/OFF'-switchable enzyme catalysis.

An artificial signal transduction model with a supramolecular recognition headgroup, a membrane anchoring group, and a pro-enzyme catalysis endgroup was constructed. The transmembrane translocation of the transducer can be reversibly regulated by competitive host-guest complexations as an input signal to control an enzyme reaction inside the lipid vesicles.

Dynamically Tunable Ultrathin Protein Membranes for Controlled Molecular Separation.

Developing the ultrathin membranes for high-performance separation still faces the challenge of both high permeance and selectivity. Herein, a large-area protein membrane was fabricated by the interfacial self-assembly of bovine serum albumin (BSA) and surfactants at the oil/water interface of emulsions. Benefiting from the ultrathin thickness and unique protein-surrounded tortuous channels, the membrane displays ultrahigh permeation flux and selective sieving capability for various molecules ranging from small dye molecules to proteins based on a dual filtration mechanism. More importantly, the rejection precision can also be reversibly regulated by the folding/unfolding transition of proteins to control the effective pore size of transport channels, even under a pressure-driven condition. This dynamically tunable ultrathin protein membrane combines the advantages of high permeance, selectivity, controllability, recyclability, and mechanical stability, which may create new opportunities for advanced applications in extended fields.

Difunctionalized pillar[5]arene-based polymer nanosheets for photodynamic therapy of Staphylococcus aureus infection.

Bacterial infections pose severe threats to global public health security. Developing antibacterial agents with both high efficiency and safety to handle this problem has become a top priority. Here, highly stable and effective polymer nanosheets have been constructed by the covalent co-assembly of a pillar[5]arene derivative and metalloporphyrin for photodynamic antibacterial therapy (PDAT). The monolayer nanosheets are strongly positively charged and thus capable of binding with Staphylococcus aureus (SA) through electrostatic interactions. Additionally, the nanosheets can be activated to generate reactive oxygen species (ROS) under white-light irradiation, and exhibit satisfactory antibacterial performance towards SA. More importantly, cell viability assays demonstrate that the nanosheets show little to no cytotoxicity impact on mammalian cells even when the concentrations are much higher than those employed in the antibacterial studies. The above results suggest that the polymer nanosheets could be an effective antibacterial agent to overcome bacterial infections and hold a broad range of potential applications in real life.

Virus-Based Supramolecular Structure and Materials: Concept and Prospects.

Rodlike and spherelike viruses are various monodisperse nanoparticles that can display small molecules or polymers with unique distribution following chemical modifications. Because of the monodisperse property, aggregates in synthetic protein-polymer nanoparticles could be eliminated, thus improving the probability for application in protein-polymer drug. In addition, the monodisperse virus could direct the growth of metal materials or inorganic materials, finding applications in hydrogel, drug delivery, and optoelectronic and catalysis materials. Benefiting from the advantages, the virus or viruslike particles have been widely explored in the field of supramolecular chemistry. In this review, we describe the modification and application of virus and viruslike particles in surpramolecular structures and biomedical research.

Construction of Ultralarge Two-Dimensional Fluorescent Protein Arrays via a Reengineered Rhodamine B-Based Molecular Tool.

The self-luminous property of enhanced green fluorescent protein (EGFP) makes it an extremely attractive building block for creating functional biomaterials. A practical challenge in the design of EGFP-based materials, however, stems from the structural and chemical heterogeneity of the EGFP surface. In this study, a maleimide-functionalized rhodamine B molecule (RhG2M) was designed as a versatile molecular tool to overcome this obstacle. Site-specific modification of an EGFP variant (EGFP-4C) with RhG2M allowed for the fabrication of a series of well-defined two-dimensional (2D) arrays that span nano- and micrometer scales. Furthermore, the resulting ultralarge 2D EGFP-4C arrays feature both structural uniformity and flexibility, together with the inherent optical properties, making them advanced materials with great potential for practical applications. In addition, this strategy can be further extended into three dimensions and applied to the modular generation of periodic functional materials with more complex structures.

Cucurbit[8]uril-based supramolecular nanocapsules with a multienzyme-cascade antioxidative effect.

A supramolecular nanocapsule was constructed by the ternary host-guest complexation of azobenzene (Azo) and methylviologen (MV) to cucurbit[8]uril (CB[8]) and the subsequent self-assembly. The supramolecular nanocapsule with both glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities can mimic the intracellular enzymatic reactive oxygen species (ROS) defense system.

Anticarcinogenic potential of gold nanoparticles synthesized from Trichosanthes kirilowii in colon cancer cells through the induction of apoptotic pathway.

Gold nanoparticles (AuNPs) is the most excellent anticancer theranostic nanoparticles synthesized through efficient, simple and green synthesis method using extracts of Trichosanthes kirilowii, extensively characterized by UV-spectroscopy, FT-IR and TEM techniques. The AuNPs, synthesized by means of T. kirilowii extracts identified that nanoparticles were ∼50 nm in size, which is an admirable nano dimension attained by green synthesis. In agreement with the outcome of microscopic cellular morphological observations, MTT assay showed effective, selective, anticarcinogenic effect of AuNPs on HCT-116 cells in a dose-dependent manner. The AuNPs significantly enhance ROS generation, cause mitochondrial membrane damage and induce morphological changes using AO/EtBr staining assay. Furthermore, AuNPs treatment induces G0/G1 phase cell-cycle arrest in HCT-116 cells. Also, AuNPs treatment activates caspase expression and downregulates the anti-apoptotic expression in HCT-116 cells. Our results point out that the phytoconsituents isolated from T. kirilowii can act as appropriate reducing and stabilizing agents in the properties of AuNPs; hereby, it leads to the green synthesis of an anti-carcinogenic agent with highly efficient potential for cancer treatment.

Computational Design and Study of Artificial Selenoenzyme with Controllable Activity Based on an Allosteric Protein Scaffold.

The establishment of new enzymatic function in an existing scaffold is a great challenge for protein engineers. In previous work, a highly efficient artificial selenoenzyme with controllable activity was constructed, based on a Ca2+ -responsive recoverin (Rn) protein. In this study, a design strategy combining docking, molecular dynamics, and MM-PBSA is presented, to predict the catalytically active site of glutathione peroxidase (GPx) on the allosteric domain of Rn. The energy contributions of the binding hot spot residues are evaluated further by energy decomposition analysis to determine the detailed substrate recognition mechanism of Rn, which provides clear guidance for artificial enzyme design for improved substrate binding (Michaelis-Menten constant, Km ).

Giant Proteinosomes As Scaffolds for Light Harvesting.

Based on an interfacial assembly strategy, a giant proteinosome was successfully fabricated by using protein-surfactant as building blocks, which formed a thin protein layer as a membrane. This approach of making protein assemblies was very facile, and it was very convenient to remove the templates of oil and get water-filled proteinosomes by dialysis. Through modifying acceptor and donor chromophores on the protein monomers, an efficient artificial light-harvesting system was successfully fabricated on the proteinosome, which was a scaffold for efficient light harvesting. Furthermore, the on-off switchable energy transfer was realized by protein folding and unfolding. The efficient artificial light-harvesting systems we designed as the potential platforms could be utilized for biomaterials.

MicroRNA-32 Regulates Development and Progression of Hepatocellular Carcinoma by Targeting ADAMTS9 and Affects Its Prognosis.

BACKGROUND MicroRNA-32 (miR-32) induces cell proliferation and metastasis in hepatocellular carcinoma (HCC), but the detailed mechanisms of miR-32 in regulating oncogenesis and development of HCC have not been clarified. The aim of this study was to investigate the effects of miR-32 on HCC and its clinical pathological significance, as well as to determine the functional connection between miR-32 and ADAMTS9 in HCC. MATERIAL AND METHODS Quantitative RT-PCR was used to assess the expression levels of miR-32 in HCC tissues, adjacent non-cancerous tissues, and liver cancer cell lines. In vitro cell proliferation, migration, and invasion assays were performed to confirm the biological functions of miR-32. Quantitative RT-PCR, Western blot analysis, and luciferase reporter assays were used to evaluate the role of miR-32 in the regulation of ADAMTS9. RESULTS miR-32 was highly expressed in HCC tissues compared with corresponding adjacent non-cancerous tissues. Over-expression of miR-32 was also found in 3 human liver cancer cell lines: SMMC-7721, Huh7, and HepG2. Moreover, increasing expression of miR-32 in HCC tissues was related to shorter overall survival. In vitro over-expression of miR-32 promoted cell proliferation, migration, and invasion; however, the under-expression of miR-32 revealed the opposite effects. Dual-luciferase reporter assay indicated that miR-32 can directly bind to the 3'-UTR of ADAMTS9. Western blot analysis showed that over-expression of miR-32 decreased expression of ADAMTS9 protein. Rescue tests further verified the connection between miR-32 and ADAMTS9. CONCLUSIONS Our data indicate that miR-32 accelerates progression in HCC by targeting ADAMTS9, and the abnormal expression of miR-32 is correlated with prognosis and could become a potential therapeutic target.

Effects of Risperidone and Aripiprazole on Serum Levels of Prolactin, Testosterone and Estradiol in Female Patients with Schizophrenia.

OBJECTIVE: To evaluate the effects of treatment with risperidone and aripiprazole on serum prolactin, testosterone and estradiol levels in female patients with schizophrenia in China. METHODS: In the retrospective study, Data were collected and included prolactin, testosterone and estradiol levels of 30 female patients with risperidone monotherapy. In the prospective study, Another 30 female schizophrenic patients were randomized to receive risperidone or adjunctive aripiprazole for six weeks. Serum prolactin, testosterone and estradiol levels were measured. RESULTS: Serum prolactin, testosterone and estradiol levels in both studies were significantly decreased after risperidone treatment compared with baseline (P<0.05), and prolactin levels remained at a high level. Serum prolactin levels in the adjunctive aripiprazole group were significantly decreased after treatment compared with baseline in the prospective study (P<0.05). Doses of 5 mg and 10 mg of adjunctive aripiprazole achieved the same efficacy at the end of treatment. CONCLUSIONS: Risperidone treatment decreased serum testosterone and estradiol levels. Adjunctive aripiprazole relieved hyperprolactinemia, but had no effect on testosterone or estradiol levels. Adjunctive aripiprazole at a dose of 5 mg is recommended for clinical use.

Cucurbit[8]uril-Based Giant Supramolecular Vesicles: Highly Stable, Versatile Carriers for Photoresponsive and Targeted Drug Delivery.

Highly stable giant supramolecular vesicles were constructed by hierarchical self-assembly of cucurbit[8]uril (CB[8])-based supra-amphiphiles for photoresponsive and targeted intracellular drug delivery. These smart vesicles can encapsulate the model drugs with high loading efficiencies and then release them by manipulating photoswitchable CB[8] heteroternary complexation to regulate the formation and dissociation of supra-amphiphiles that cause dramatic morphological changes of the assemblies to achieve remote optically controlled drug delivery. More importantly, the confocal microscopy analysis, cellular uptake experiment, and cell viability assay have shown that the giant vesicles are able to maintain the structural integrity and stability within actual cellular environments and exhibit obvious advantages for intracellular drug delivery such as low toxicity, easy surface modification for tumor-targeting selectivity, and rapid internalization into different human cancer cell lines. A synergistic mechanism that integrates multiple pathways including energy-dependent endocytosis, macropinocytosis, cholesterol-dependent endocytosis, and microtubule-related endocytosis was determined to facilitate the internalization process. Moreover, cytotoxicity experiments and flow cytometric analysis have demonstrated that the doxorubicin hydrochloride-loaded vesicles exhibited a significant therapeutic effect for tumor cells upon UV light irradiation, which makes the photoresponsive system more promising for potential applications in pharmaceutically relevant fields.

[Association between -590C/T polymorphisms of interleukin-4 gene and chronic periodontitis: a meta-analysis].

PURPOSE: To evaluate the relationship between -590C/T polymorphisms of interleukin-4 gene and chronic periodontitis using meta-analysis. METHODS: The selected studies were pooled from eight major electronic databases for case-control study. To gain a more precise estimation of the relationship, a stratified meta-analysis with two subgroups was performed according to the races. Heterogeneity, publication bias and sensitivity analysis were also explored. RESULTS: Totally 8 studies were recruited. Total sample sizes for chronic periodontitis and control groups were 628 and 717, respectively. Meta-analysis showed that both -590C/T polymorphisms and allele frequency were not significantly associated with chronic periodontitis. In subgroup analysis, a significant association of increased chronic periodontitis risk and T allele was found. The results also indicated a significant correlation between -590C/T polymorphisms of IL-4 and Caucasian who suffered from chronic periodontitis(C vs.T: OR=0.71,95% CI=0.56-0.89;CC vs.CT:OR=0.60,95% CI=0.38-0.94;recessive genetic model CC vs.CT+TT:OR=0.61,95% CI=0.42-0.88), further analysis of the results showed the CC genotype was about 39% less likely to have chronic periodontitis than the CT and TT genotype in Caucasian individuals.However, these significant associations was not found in Asian group. CONCLUSIONS: The meta-analysis suggests there may be an important effect of single nucleotide polymorphisms (SNPs) in the promoter region of IL-4 gene on the pathogenesis of chronic periodontitis in Caucasian. This warrants further investigation in larger studies and multi-race epidemiological to evaluate the results.

[Meta analysis on effect of local application of chlorhexidine with scaling and root planing].

PURPOSE: Meta analysis was used to assess the effect of local drug delivery system of chlorhexidine (CHX) as an adjunct to scaling and root planing(SRP) versus SRP alone in patients with chronic periodontitis. METHODS: The selected studies of randomized controlled trials(RCTs) were pooled from six major electronic database up to May,2009 as CHX plus SRP versus SRP with at least 3 months of follow-up. Several English full texts were hand searched. Outcome measures were probing depth(PD) reduction and clinical attachment level (CAL)gain. RESULTS: Seven studies that met inclusion criteria were entered into the Meta analysis. A significant mean reduction of PD in patients with CHX plus SRP was observed, but there were no significant difference in CAL between the two treatment groups. CONCLUSIONS: Chlorhexidine as an adjunct to SRP could reduce probing depth in the treatment of chronic periodontitis. Supported by Research Project of Education Bureau of Liaoning Province(Grant No.20061021).