Zwitterionic Polysulfone Copolymer/Polysulfone Blended Ultrafiltration Membranes with Excellent Thermostability and Antifouling Properties.

Membrane fouling has been one of the most important challenges in membrane separation operations. In this study, we report a facile strategy to prepare antifouling polysulfone (PSf) UF membranes by blending amphiphilic zwitterion polysulfone-co-sulfobetaine polysulfone (PSf-co-SBPSf) copolymer. The copolymer chemical structure was characterized by 1HNMR spectroscopy. The PSf/PSf-co-SBPSf blend membranes with various zwitterionic SBPSf segment contents exhibited better surface hydrophilicity and excellent antifouling ability compared to PSf and PSf/PEG membranes. The significant increase of both porosity and water permeance indicates that the PSf-co-SBPSf has a pore-forming effect. The pure water flux and flux recovery ratio of the PSf/PSf-co-SBPSf blend membranes were both remarked to improve 286.43 L/m2h and 92.26%, while bovine serum albumin (BSA) rejection remained at a high level (97.66%). More importantly, the water flux and BSA rejection see minimal variance after heat treatment, indicating excellent thermostability. Overall, the PSf/PSf-co-SBPSf blend membranes achieved a comprehensive performance of sustainable hydrophilic, high permeation flux, and remarkable antifouling ability, thus becoming a promising candidate in high-temperature separation application.

Is there a relationship between neural EGFL like 1 (NELL1) promoter hypermethylation and prognosis of gastric cancer?

We hypothesized that neural EGFL like 1 (NELL1) promoter hypermethylation might be associated with the prognosis of gastric cancer. Some studies considered NELL1 as a tumor suppressor gene and our research confirmed for the first time the hypermethylation in the promoter region of NELL1 by the application of mass spectrometry. Promoter hypermethylation can cause the silencing of tumor suppressor genes and promote tumor progression. Based on present studies and research results, we proposed that NELL1 promoter hypermethylation might be associated with cancer staging and the survival of gastric cancer patients and had prognostic value. We hoped that NELL1 promoter hypermethylation would be applied not only for early detection but also prognosis prediction of gastric cancer and would become a new prognostic biomarker.

Study on the Core-Shell Reversion of PSBMA- b-PLMA Nanoparticles for the Fabrication of Antifouling Coatings.

In this paper, two series of poly(sulfobetaine methacrylate)- b-poly(lauryl methacrylate) (PSBMA- b-PLMA) diblock copolymers were prepared to investigate the core-shell reversion of amphiphilic copolymers. Experimental results proved that the PSBMA- b-PLMA copolymers can be self-assembled as core-shell nanoparticles in chloroform. Moreover, 1H NMR spectra and contact angle measurements revealed that there is a transitional PSBMA/PLMA block ratio of 0.6, above which the nanoparticles are capable of switching their core and shell in aqueous solution. Consequently, nanoparticles with PSBMA/PLMA block ratios above 0.6 showed superior antifouling and antibacterial abilities to those with block ratios below 0.4. Moreover, it was also found that the block chain length plays an important role in core-shell reversion as evidenced by 1H NMR spectra, water contact angle, and antifouling tests. As a result, coatings fabricated with the PLMA100 series of nanoparticles showed better antifouling abilities than those of the PLMA150 series at the same block ratio probably because of the thinner shell of PLMA100 copolymers. PSBMA100- b-PLMA100 was proved to be the best candidate for the fabrication of antifouling coatings as it exhibited the highest efficacy in antibacterial adhesion and antiprotein adsorption. This study provided a facile method to fabricate antifouling coatings by developing amphiphilic diblock copolymers with tuned hydrophobic/hydrophilic block ratio, block chain length, etc.

Is co-expression of USP22 and HSP90 more effective in predicting prognosis of gastric cancer?

The ubiquitin-specific peptidase 22 (USP22) belongs to the largest subfamily of deubiquitylases and recent studies indicate that overexpression of USP22 may promote gastric cancer progression and predict prognosis. But little is known about the interaction network of USP22 in gastric cancer. In this study, we applied bioinformatics methods and found that USP22 was correlated with the heat shock protein 90 (HSP90) which is now considered to be a biomarker to predict the prognosis of gastric cancer. Then the siRNA transfection and western blotting were used to testify the correlation of USP22 and HSP90 in gastric cancer cells. The immunohistochemistry staining of the microarrays was applied to confirm the correlation of USP22 and HSP90 expression in gastric cancer tissue and further analysis showed that co-expression of USP22 and HSP90 was related to lymph node metastasis and more effective in predicting the prognosis of gastric cancer. In summary, our data demonstrate that correlation exists between USP22 and HSP90 expressions in gastric cancer and co-expression of USP22 and HSP90 may be more effective in predicting prognosis of gastric cancer.

Creation of antifouling microarrays by photopolymerization of zwitterionic compounds for protein assay and cell patterning.

Nonspecific binding or adsorption of biomolecules presents as a major obstacle to higher sensitivity, specificity and reproducibility in microarray technology. We report herein a method to fabricate antifouling microarray via photopolymerization of biomimetic betaine compounds. In brief, carboxybetaine methacrylate was polymerized as arrays for protein sensing, while sulfobetaine methacrylate was polymerized as background. With the abundant carboxyl groups on array surfaces and zwitterionic polymers on the entire surfaces, this microarray allows biomolecular immobilization and recognition with low nonspecific interactions due to its antifouling property. Therefore, low concentration of target molecules can be captured and detected by this microarray. It was proved that a concentration of 10ngmL-1 bovine serum albumin in the sample matrix of bovine serum can be detected by the microarray derivatized with anti-bovine serum albumin. Moreover, with proper hydrophilic-hydrophobic designs, this approach can be applied to fabricate surface-tension droplet arrays, which allows surface-directed cell adhesion and growth. These light controllable approaches constitute a clear improvement in the design of antifouling interfaces, which may lead to greater flexibility in the development of interfacial architectures and wider application in blood contact microdevices.

Affinity Monolith-Integrated Microchips for Protein Purification and Concentration.

Affinity chromatography is a valuable method to purify and concentrate minute amount of proteins. Monoliths with epoxy groups for affinity immobilization were prepared by direct in-situ photopolymerization of glycidyl methacrylate and ethylene glycol dimethacrylate in porogenic solvents consisting of 1-dodecanol and cyclohexanol. By integrating affinity monoliths onto a microfluidic system, targeted biomolecules can be captured and retained on affinity column, while other biomolecules having no specific interactions toward the immobilized ligands flow through the microchannel. Therefore, proteins which remain on the affinity column are purified and concentrated, and then eluted by appropriate solutions and finally, separated by microchip capillary electrophoresis. This integrated microfluidic device has been applied to the purification and separation of specific proteins (FITC-labeled human serum albumin and IgG) in a mixture.

MALDI-TOF Mass Array Analysis of Nell-1 Promoter Methylation Patterns in Human Gastric Cancer.

Mass spectrometry (MS) enables rapid and sensitive qualitative and quantitative analyses of biomolecules (proteins, peptides, oligosaccharides, lipids, DNA, and RNA), drugs, and metabolites. MS has become an essential tool in modern biomedical research, including the analysis of DNA methylation. DNA methylation has been reported in many cancers, suggesting that it can be utilized as an early biomarker to improve the early diagnosis rate. Using matrix-assisted laser desorption/ionization time-of-flight MS and MassCLEAVE reagent, we compared Nell-1 hypermethylation levels among tumor tissues, paracarcinoma tissues, and normal tissues from gastric cancer patients. Almost 80% of the CpG sites in the amplicons produced were covered by the analysis. Our results indicate a significant difference in methylation status between gastric cancer tissue (a higher level) and normal tissue. The same trend was identified in gastric cancer tissue versus paracarcinoma tissue. We also detected lower relative expression of Nell-1 by real-time PCR. Furthermore, immunohistochemical analyses revealed that Nell-1 staining was less intense in cancer tissue relative to normal tissue and that the tumor cells had spread to the muscle layer. These findings may serve as a guide for the early diagnosis of gastric cancer.

S-1 combined with cisplatin versus cisplatin alone for the treatment of advanced gastric cancer: a pilot randomized-controlled trial.

We aimed to assess the efficacy and safety of S-1 combined with cisplatin (SC) over cisplatin alone (C) for the treatment of advanced gastric cancer in China. Between July 2009 and June 2011, 72 eligible patients with advanced gastric cancer were selected and divided randomly into two groups. Thirty-six patients received SC, with S-1 on days 1 through 14 of a 21-day cycle and cisplatin (60 mg/m on day 1) every 4 weeks for two cycles. The other 36 patients were administered only cisplatin (in the same manner as SC). The primary outcome was overall survival. The secondary outcomes were progression-free survival and adverse events. The 2-year overall response rate was 51.5 and 42.3% for the SC and C groups, respectively, and the difference was statistically significant, whereas the median overall survival was 9.4 months (range, 1.9-24.4 months) and 7.6 months (range, 1.7-21.4 months), respectively (P=0.039). The median progression-free survival was 7.7 months for SC (range, 1.8-19.4 months), whereas it was 6.5 months (range, 1.5-16.4 months) for C (P=0.047). The toxicity profile was similar in both groups. In summary, we have shown that S-1 combined with cisplatin is more effective, with acceptable toxicity in comparison with cisplatin alone in Chinese patients with advanced gastric cancer. Chinese Clinical Trials Register: ChiCTR-TRC-13003993.

Immunopathogenesis of chronic hepatitis B.

Chronic hepatitis B (CHB) is a widespread infectious disease with unfavorable outcomes and life-threatening consequences for patients, in spite of modern vaccination and antiviral treatment modalities. Cutting-edge experimental approaches have demonstrated key pathways that involve cross-talk between viral particles and host immune cells. All events, including penetration of hepatitis B virus (HBV) particles into host cells, establishing persistence, and chronization of CHB infection, and possibility of complete elimination of HBV particles are controlled by the immune system. Researchers have paid special attention to the replication capacity of HBV in host cells, which is associated with cellular changes that reflect presentation of viral antigens and variability of HBV antigen features. In addition, specific HBV proteins have an immune-modulating ability to initiate molecular mechanisms that "avoid" control by the immune system. The relationship between immunological shifts and chronic infection stages has been intensively studied since it was recognized that the immune system is a direct participant in the recurrent (cyclic) nature of CHB. Understanding the wide diversity of molecular pathways and the crosstalk between innate and adaptive immune system components will provide fresh insight into CHB immune pathogenesis and the possibilities of developing new treatment strategies for this disease.

Fluorescent measurement of affinity binding between thrombin and its aptamers using on-chip affinity monoliths.

A microfluidic chip with integrated 2mm long monoliths incorporated with poly(ethylene glycol) (PEG) groups was developed for thrombin-aptamer interaction study. The non-G quartet forming oligonucleotide coated monoliths was compared to a 15 mer thrombin-binding aptamer, in which affinity binding and elution processes were real-time monitored fluorescently. The results showed that the fluorescence intensity of aptamer stationary phase is approximately 10 times higher than that of the control column, which is probably due to the successful suppression of nonspecific adsorption between thrombin and aptamers/monoliths by using PEG-monolith. The experiment was repeated using human serum albumin (HSA) and green fluorescence protein (GFP) as interferences, it was double confirmed that thrombin was selectively retained by PEG-monolith. An elution efficiency of 75% was achieved with an elute of 200mM acetic acid and 2M NaCI, and the eluted thrombin was successfully separated in an ionic buffer system of 20mM NaHCO3 (pH 9.5) with 3% PEG. The hydrophilic and antifouling properties of PEG-monolith greatly decrease nonspecific adsorption and enhance detection sensitivity, which provided an alternative method to perform on-chip fluorescent measurement of bioaffinity binding.

Fabrication of two-layer poly(dimethyl siloxane) devices for hydrodynamic cell trapping and exocytosis measurement with integrated indium tin oxide microelectrodes arrays.

The design, fabrication and test of a microfluidic cell trapping device to measure single cell exocytosis were reported. Procedures on the patterning of double layer template based on repetitive standard photolithography of AZ photoresist were investigated. The replicated poly(dimethyl siloxane) devices with 2.5 µm deep channels were proved to be efficient for stopping cells. Quantal exocytosis measurement can be achieved by targeting single or small clumps of chromaffin cells on top of the 10 µm × 10 µm indium tin oxide microelectrodes arrays with the developed microdevice. And about 72 % of the trapping sites can be occupied by cells with hydrodynamic trapping method and the recorded amperometric signals are comparable to the results with traditional carbon fiber microelectrodes. The method of manufacturing the microdevices is simple, low-cost and easy to perform. The manufactured device offers a platform for the high throughput detection of quantal catecholamine exocytosis from chromaffin cells with sufficient sensitivity and broad application.

Erythropoietin protects against cisplatin-induced nephrotoxicity by attenuating endoplasmic reticulum stress-induced apoptosis.

BACKGROUND: The anticancer drug cisplatin (CP)-induced nephrotoxicity associated with apoptosis plays crucial roles in tumor patients. Erythropoietin (EPO) has recently been shown to enhance recovery from CP-induced acute kidney injury (AKI) in rats by exerting anti-apoptotic effects. However, the molecular mechanisms of Erythropoietin protects against CP-induced AKI are not very clear. The present study investigated the protective effects of erythropoietin (EPO) against CP-induced nephrotoxicity and the possible mechanism in rats. METHODS: Sprague-Dawley (SD) rats were randomly divided into four groups: (1) Control group (n=16): which received a single intraperitoneal injection of vehicle (0.9% saline; 5 mL/kg); (2) CP group (n=16): which received a single intraperitoneal injection of 10.0 mg/kg CP (previously dissolved at 2.0 mg/mL in 0.9% saline solution); (3) CP+rHuEPO group (n=16): which received rHuEPO (5000 U/kg) with co-injection of the LY294002 vehicle dimethyl sulfoxide (DMSO, 33.3 µL/kg; Sigma, St. Louis, MO, USA) by tail vein injection 2 days before CP administration, 15 min before CP administration, and 2 days after CP administration; (4) CP+rHuEPO+LY group (n=16): which received LY294002 (0.3 mg/kg) 10 min before rHuEPO administration by three injections into the tail vein at 2-day intervals beginning 2 days before CP administration. Apoptosis was assessed by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method. The expressions of C/EBP-homologous protein (CHOP), glucose-regulated protein 78 (GRP78), caspase-12, the phosphorylation of Akt, cleaved-caspase-3 and EPO receptor (EPOR) were measured after CP-treated. In addition, light microscopy and immunohistochemistry examinations were performed. RESULTS: The levels of serum urea and creatinine were increased at 96 h after CP-administered group. The eHuEPO-administered group had significantly lower serum creatinine levels. CP caused an increase in TUNEL-positive cells that was accompanied and apoptotic cell death induced by CP was significantly abrogated by rHuEPO at 96h by morphological evidence of apoptosis. The over-expression of CP-induced endoplasmic reticulum (ER) stress markers (CHOP and GRP78) and activation of caspase-12 were suppressed by rHuEPO, which also attenuated the CP-induced suppression of phosphatidylinositol-3kinase/Akt (PI3K/Akt) signaling in rat kidneys. In addition, LY294002 diminished the effect of rHuEPO on renal protection and antiapoptosis. CONCLUSIONS: Injection of rHuEPO enhance recovery from CP-induced AKI in rats by ameliorating renal functional impairment and exerting important anti-apoptotic effects. However, rHuEPO inhibited CP-induced AKI by a possible mechanism involving PI3K/Akt activation and the inhibition of ER stress-mediated apoptosis.

Overexpression of the HIF hydroxylase PHD3 is a favorable prognosticator for gastric cancer.

Hypoxia-induced factors (HIFs) play a central role in the adaptive mechanisms of cancer cells to survive under conditions of hypoxia. HIFs are regulated by prolyl hydroxylases (PHDs) among which PHD3 is implicated as a tumor suppressor. We aimed to correlate PHD3 expression with clinicopathologic parameters and to evaluate its prognostic significance in gastric cancer. The 101 tissue samples were collected from 83 resected stages I­IV gastric cancer patients, which were grouped as non-cancerous mucosa (n=18) and primary carcinoma (n=83). PHD3 expression was evaluated by immunohistochemistry. We adopted Pearson chi-square test, univariate analysis, multivariate analysis and Kaplan­Meier method. The positive frequency of PHD3 in cancer cells was 42.2%, whereas non-cancerous mucosa had no detectable PHD3. The expression of PHD3 increased significantly from non-cancerous mucosa to cancer. A significant difference was observed between PHD3 expression and tumor differentiation (P=0.007). The overexpression of PHD3 was associated with well differentiation. In univariate analyses, American Joint Committee on Cancer (AJCC) stage (P<0.0001), pT classification (P<0.0001), pN classification (P<0.0001), differentiation (P=0.0121), peritoneal metastasis (P=0.0006) and gross features (P=0.0104) were significantly associated with survival except PHD3 (P=0.2228) (Table 3). In multivariate analysis, AJCC stage was prognostically independent [hazard ratio (HR), 3.078; 95% confidence interval (CI), 2.228­4.252; P<0.0001]. Overexpression of PHD3 is a favorable prognosticator for gastric cancer. AJCC stage is an independent prognostic factor of gastric cancer.

The level of serum bilirubin associated with coronary lesion types in patients with coronary artery disease.

BACKGROUND: Serum bilirubin has been proven to be associated with coronary artery disease (CAD). However, how serum bilirubin is related to the complexity of coronary artery lesions is still unknown. METHODS AND RESULTS: One thousand two hundred and sixty patients (men 775, 61.5%, mean age, 59.3  ±â€Š 8.2 years) diagnosed with unstable angina were enrolled in the study. Patients were categorized into three major groups and group III was further divided into four subgroups according to the guidelines of AHA/ACC 1993 described in the Methods section. The total serum bilirubin levels showed significant differences among the three major groups (group I vs. group II, 14.8 ± 5.8 vs. 13.7 ± 4.7 µmol/l, P=0.017; group I vs. group III, 14.8 ± 5.8 vs. 12.6 ±  4.4 µmol/l, P<0.001; group II vs. group III, 13.7 ± 4.7 vs. 12.6 ± 4.4 µmol/l, P=0.009). The difference was further seen among the subgroups. Logistic regression analysis demonstrated that age, male sex, histories of hypertension and diabetes, and total serum bilirubin were independent risk factors for CAD. However, in the subgroups, only age, male sex, history of hypertension and total serum bilirubin were associated with CAD. Total serum bilirubin showed the strongest relationship (odds ratio=0.95, 95% confidence interval 0.91-0.98, P=0.001). CONCLUSION: Total serum bilirubin level is an independent risk factor for CAD. It has a strong relationship with coronary artery lesion types.

Cystatin C levels are associated with the prognosis of systolic heart failure patients.

BACKGROUND: Cystatin C, which has long been regarded as a biomarker that indicates kidney functions, has recently been recognized as an inflammatory marker in the human body. AIM: To elucidate how cystatin C is related to the prognosis of systolic heart failure patients. METHODS: Patients with systolic heart failure who were admitted to the fourth affiliated hospital of Harbin Medical University between January and April 2008 were enrolled in this study. Serum homocysteine, high-sensitivity C-reactive protein (hs-CRP) and cystatin C levels were determined and all the patients received an average of 2 years of follow-up for occurrence of death, heart transplantation or readmission with worsening heart failure. RESULTS: Of 138 patients enrolled, those who experienced adverse outcomes (e.g. cardiac death, heart transplantation or progressive heart failure) (n = 21) had considerably higher mean levels of serum homocysteine (28.6 ± 13.4 vs 14.4 ± 6.3mg/L; P < 0.01), hs-CRP (17.5 ± 14.1 vs 6.4 ± 7.7 µmol/L; p < 0.01) and cystatin C (1.63 ± 0.81 vs 0.91 ± 0.27 mg/L; P < 0.01) than those without adverse outcomes (n = 117). Furthermore, the Cox proportional hazards model demonstrated that serum homocysteine, hs-CRP and cystatin C are all independent predictors of adverse outcomes. CONCLUSIONS: Cystatin C, together with hs-CRP and homocysteine, is an independent risk factor that is important in the prognosis of patients with systolic heart failure.

Imaging and cell targeting characteristics of magnetic nanoparticles modified by a functionalizable zwitterionic polymer with adhesive 3,4-dihydroxyphenyl-l-alanine linkages.

Multifunctional magnetic nanoparticles (MNPs) modified by a zwitterionic polymer (pCBMA-DOPA(2)) containing one poly(carboxybetaine methacrylate) (pCBMA) chain and two 3,4-dihydroxyphenyl-L-alanine (DOPA) residue groups were developed. Results showed that MNPs modified by pCBMA were not only stable in complex media, but also provided abundant functional groups for ligand immobilization. The pCBMA-DOPA(2) MNPs had a hydrodynamic particle size of about 130 nm, a strong saturation magnetization of 110.2 emu/g Fe and a high transverse relaxivity of 428 mM(-1)s(-1). Long-term stability in phosphate-buffered saline (PBS) and 10% NaCl solution was achieved for over six months. Compared to MNPs coated with dextran, pCBMA-DOPA(2) MNPs presented better stability in 100% human blood serum at 37 degrees C. Macrophage cell uptake studies revealed that the uptake ratio of pCBMA-DOPA(2) MNPs was much lower than that of dextran MNPs. Furthermore, quantitative analysis results showed that after pCBMA-DOPA(2) MNPs were conjugated with a targeting RGD peptide, uptake by human umbilical vein endothelial cell (HUVEC) was notably increased, which was further visualized by magnetic resonance imaging (MRI).

Ultra-low fouling and functionalizable zwitterionic coatings grafted onto SiO2 via a biomimetic adhesive group for sensing and detection in complex media.

Non-specific protein binding from human plasma and serum has severely hindered the full capabilities of biosensors concerned with cancer biomarker detection. Currently, there is a strong desire for developing new materials which allow for the convenient attachment of an ultra-low fouling and functionalizable surface coating which can be used for highly sensitive and label-free detection of target analytes directly from complex media. In this work, a short 20 min in situ "graft to" protocol using Tris pH 8.5 buffer was developed for zwitterionic carboxybetaine methacrylate (CBMA) polymer conjugates containing the adhesive biomimetic moiety, 3,4-dihydroxy-L-phenylalanine (DOPA), on SiO(2) substrates. Using a surface plasmon resonance (SPR) biosensor, different buffers, pH values, salt concentrations, and temperatures were investigated for determining the "graft to" conditions that yield dense polymer films which both minimize non-specific protein adsorption and maximize antibody immobilization. The optimized surface coatings were shown to be highly protein resistant to 100% human blood plasma and serum. Subsequent antibody functionalized surfaces without any blocking agents enabled the specific detection of the cancer biomarker ALCAM directly from undiluted human serum down to 64 ng/mL. The successful use of this zwitterionic surface coating for detection from complex media on SiO(2) surfaces indicates its potential for broad impacts in the development of implantable medical devices, in vivo diagnostics, and nano-scale biosensors.

Functionalizable and ultra-low fouling zwitterionic surfaces via adhesive mussel mimetic linkages.

In this work, a biomimetic polymer (pCB(2)-catechol(2)), with two zwitterionic poly(carboxybetaine) (pCB) arms for ultra-low fouling and two adhesive catechol groups for surface anchoring, was developed. Two pCB arms were grown from an initiator with two catechol groups via atom transfer radical polymerization (ATRP). Binding tests of pCB(2)-catechol(2) were performed on a gold surface under a range of conditions such as pH values and solvents. Protein adsorption from single protein solutions of fibrinogen and lysozyme, and complex media of 100% blood plasma and serum was evaluated using a surface plasmon resonance (SPR) sensor. Results are compared with those from two other polymers (i.e., one polymer with one pCB chain and one catechol group, termed as pCB-catechol, and another polymer with one pCB chain and two catechol groups, termed as pCB-catechol(2)). Furthermore, the direct immobilization of anti-activated leukocyte cell adhesion molecule (anti-ALCAM) was carried out on the pCB(2)-catechol(2) modified surface. Results showed that the antibody-immobilized surface maintained its excellent ultra-low fouling properties. The detection of activated leukocyte cell adhesion molecule (ALCAM) in 100% blood plasma with high sensitivity and specificity was achieved. This work demonstrates an effective and convenient strategy to obtain functionalizable and ultra-low fouling surfaces.

Aptamer-encoded nanopore for ultrasensitive detection of bioterrorist agent ricin at single-molecule resolution.

The molecular-scale pore structure, called nanopore, can be formed from protein ion channels by genetic engineering or fabricated on solid substrates using fashion nanotechnology. Target molecules in interaction with the functionalized lumen of nanopore, can produce characteristic changes in the pore conductance, which act as fingerprints, allowing us to identify single molecules and simultaneously quantify each target species in the mixture. Nanopore sensors have been created for tremendous biomedical detections, with targets ranging from metal ions, drug compounds and cellular second messengers, to proteins and DNAs. Here we will review our recent discoveries with a lab-in-hand glass nanopore: single-molecule discrimination of chiral enantiomers with a trapped cyclodextrin, and sensing of bioterrorist agent ricin.