MCF-7 cell - derived exosomes were involved in protecting source cells from the damage caused by tributyltin chloride via transport function.

Tributyltin chloride (TBTC) is a ubiquitous environmental pollutant with various adverse effects on human health. Exosomes are cell - derived signaling and substance transport vesicles. This investigation aimed to explore whether exosomes could impact the toxic effects caused by TBTC via their transport function. Cytotoxicity, DNA and chromosome damage caused by TBTC on MCF-7 cells were analyzed with CCK-8, flow cytometry, comet assay and micronucleus tests, respectively. Exosomal characterization and quantitative analysis were performed with ultracentrifugation, transmission electron microscope (TEM) and bicinchoninic acid (BCA) methods. TBTC content in exosomes was detected with Liquid Chromatography-Mass Spectrometry (LC-MS). The impacts of exosomal secretion on the toxic effects of TBTC were analyzed. Our data indicated that TBTC caused significant cytotoxicity, DNA and chromosome damage effects on MCF-7 cells, and a significantly increased exosomal secretion. Importantly, TBTC could be transported out of MCF-7 cells by exosomes. Further, when exosomal secretion was blocked with GW4869, the toxic effects of TBTC were significantly exacerbated. We concluded that TBTC promoted exosomal secretion, which in turn transported TBTC out of the source cells to alleviate its toxic effects. This investigation provided a novel insight into the role and mechanism of exosomal release under TBTC stress.

A sensitive lateral flow test strip sensor for visual detection of acid red 18 in food using bicentric-emission carbon dots.

Hazardous synthetic colorants have found widespread use in food production, and excessive consumption of these pigments can pose potential risks to human health. In this study, we propose an ultrasensitive fluorescence method for the analysis of Acid Red 18 (AR18) in food products. The method is based on the nitrogen-doped carbon dots (N-CDs) derived from tris and resorcinol through a hydrothermal way. The as-synthesized N-CDs exhibit two emission centers at 425 nm and 541 nm, corresponding to the excitation wavelengths of 377 nm and 465 nm, respectively. Upon the addition of AR18, the fluorescence intensity at 541 nm significantly decreases with a simultaneous, though less pronounced, reduction in the intensity at 425 nm, which is attributed to the localization of fluorescence resonance energy transfer (L-FRET). Specifically, a novel ratiometric fluorescent probe was constructed based on the extracted data from the 3D fluorescence excitation-emission matrix. This probe demonstrates a wide linear range from 0.0539 to 30 µM and a low limit of detection (LOD) of 53.9 nM. For practical applications, a portable fluorescent sensor based on a lateral flow test strip (LFTS) was designed for real-time monitoring of AR18. Color channel values were determined using a smartphone application, resulting in a satisfactory LOD of 75.3 nM. Furthermore, the suitability of the proposed ratiometric fluorescent probe was validated through the detection of AR18 in real food samples, consistently achieving recovery rates in the range of 99.7-101.4%. This research not only expands the scope of CDs in sensing fields, but also provides an effective strategy for the development of an excellent platform for real-time AR18 detection, contributing to public food safety.

Preparation of Multifunctional Nano-Molybdenum Disulfide and Its Tribological Properties in Water-Based Cutting Fluids.

In order to meet the advanced requirements of the manufacturing industry, the use of water-based cutting fluids (WCFs) in metal processing is gradually increasing. However, their lubrication performance still needs to be improved considerably. Therefore, new multifunctional molybdenum disulfide nanoparticles (m-MoS2 NPs) were developed to improve the lubricating properties of WCFs. M-MoS2 NPs modified with silver nanoparticles were prepared by an in situ surface modification. The morphology and chemical composition of the m-MoS2 NPs were characterized by scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HR-TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). Furthermore, the dispersion and bactericidal properties of m-MoS2 NPs with different weight percents in WCFs were also studied experimentally. The effect of m-MoS2 NPs concentration on friction properties and their friction mechanism were investigated in this research. The results revealed that the prepared m-MoS2 NPs were all nanoscale particles with a layered structure. The dispersion and bactericidal properties of m-MoS2 NPs in WCFs were better than those of MoS2 NPs. The best dispersion and bactericidal properties were observed with 1 wt % MoS2 NPs, as well as friction reduction and antiwear properties. During friction, the two friction surfaces were in the boundary lubrication state,and the prepared m-MoS2 NPs entered the friction contact zone along with the WCFs. A friction chemical reaction film rich in MoS2 and Ag NPs was formed on the friction surface to fill and repair the worn surface, exerting a good lubrication effect.

The E3 ubiquitin ligase CSIT1 regulates critical sterility-inducing temperature by ribosome-associated quality control to safeguard two-line hybrid breeding in rice.

Two-line hybrid breeding can fully utilize heterosis in crops. In thermo-sensitive genic male sterile (TGMS) lines, low critical sterility-inducing temperature (CSIT) is vital to safeguard the production of two-line hybrid seeds in rice (Oryza sativa), but the molecular mechanism determining CSIT is unclear. Here, we report the cloning of CSIT1, which encodes an E3 ubiquitin ligase, and show that CSIT1 modulates the CSIT of thermo-sensitive genic male sterility 5 (tms5)-based TGMS lines through ribosome-associated quality control (RQC). Biochemical assays demonstrated that CSIT1 binds to the 80S ribosomes and ubiquitinates abnormal nascent polypeptides for degradation in the RQC process. Loss of CSIT1 function inhibits the possible damage of tms5 to the ubiquitination system and protein translation, resulting in enhanced accumulation of anther-related proteins such as catalase to suppress abnormal accumulation of reactive oxygen species and premature programmed cell death in the tapetum, thereby leading to a much higher CSIT in the tms5-based TGMS lines. Taken together, our findings reveal a regulatory mechanism of CSIT, providing new insights into RQC and potential targets for future two-line hybrid breeding.

Construction of fluorescent sensor array and three-dimensional microfluidic paper based analytical device for specific identification and visual determination of antibiotics in food.

For antibiotics misuse since the global outbreak of COVID 19, a novel strategy for discriminating and detecting antibiotics is proposed based on the graphene quantum dots with multi-doped heteroatoms including F, N and P (M-GQDs), which exhibit blue emission (419.0 nm) under the excitation of 336.0 nm. Specifically, the fluorescence of M-GQDs is quenched by tetracyclines (TCs) owing to inner filter effect (IFE) and enhanced by alkane-modified fluoroquinolones (AFQs), which is attributed to restricted conformational rotation based on π-π stacking, hydrogen-bonding and electrostatic interactions. Meanwhile, the electron-accepting property of oxazine ring in oxazine-modified fluoroquinolones (OFQs) increases emission peak at 498.0 nm and decreases emission peak at 419.0 nm as the color changes from blue to cyan. Moreover, a cascade system integrated with 3D microfluidic paper-based analytical device (3D-µPAD) is applied successfully for visually distinguishing three antibiotics, which shows great potential and versatility of M-GQDs for food safety monitoring.

Aggregation-induced bimodal excitation of nitrogen-doped carbon dots for ratiometric sensing of new coccine and solid-state multicolor lighting.

Trace detection of foodstuff pigments have gained increasing attention because of their close association with biological and environmental processes. Herein, we propose an innovative bimodal excitation nitrogen-doped carbon dots (N-CDs) for ratiometric sensing of new coccine (NC) pigment, which are synthesized by using melamine and o-phenylenediamine as precursors via solvothermal treatment. With the increase of the N-CDs concentration, N-CDs exhibit not only a concentration-dependent tunable color behavior, but also a novel aggregation-induced bimodal excitation phenomenon. Considering this distinctive bimodal excitation behavior, a ratiometric sensor based on N-CDs has been developed for the detection of the NC in different organic solvents due to the inner filter effect and fluorescence resonance energy transfer. The intensity ratio of two excitation signals is linear with the NC concentration in the range of 0.032-100 µM, and the limit of detection is as low as 32.1 nM. Meanwhile, we realize the design of multicolor-emission N-CDs/polymer films. All in all, this work presents a novel kind view of the mechanism of distinctive bimodal excitation of N-CDs, and further proposes an innovative ratiometric method for the screening analysis of NC in food samples and environmental pollutants.

Construction of graphene quantum dots ratiometric fluorescent probe by intermolecular electron transfer effect for intelligent and real-time visual detection of ofloxacin and its L-isomer in daily drink.

The design of intelligent and real-time sensing devices is significant in the medical drug monitoring field, but it is still highly challenging. Here, ratiometric fluorescent detections of ofloxacin (OFL) and its L-isomer levofloxacin (LEV) constructed from tri-doped graphene quantum dots (T-GQDs) are reported, and the detection limits reach as low as 46/67 nM toward OFL/LEV due to the intermolecular electron transfer (intermolecular ET) effect. After adding OFL/LEV, the generation of electrostatic bond provides a channel for the intermolecular ET from the edge of T-GQDs to OFL/LEV, resulting in the fluorescence quenching at 414 nm and the fluorescence promoting at 498 nm. Furthermore, a smartphone can be used for the visual and quantitative detection of OFL and LEV by identifying the RGB values of test paper and drink samples. This work not only reveals the physics mechanism of ratiometric detection, but also develops a convenient smartphone diagnostic for OFL and LEV.

Activated cascade effect for dual-mode ratiometric and smartphone-assisted visual detection of curcumin and F- based on nitrogen-doped carbon dots.

The growing persistence of harmful ion or drug molecular residues has always been considered as a matter of concern due to its importance in biological and environmental processes, which requires taking measures to maintain environmental health sustainably and effectively. Inspired by the multi-system and visual quantitative detection of nitrogen-doped carbon dots (N-CDs), we develop a novel cascade nano-system based on dual emission carbon dots for on-site visual quantitative detection of curcumin and fluoride ion (F-). Herein, tris (hydroxymethyl) aminomethane (Tris) and m-dihydroxybenzene (m-DHB) are elected as reaction precursors to synthesize dual-emission N-CDs by a one-step hydrothermal method. The obtained N-CDs exhibit dual emission peaks at 426 nm (blue) and 528 nm (green) with quantum yields of 53 % and 71 %, respectively. Then, trace curcumin and F- intelligent off-on-off sensing probe is formed by taking advantage of the activated cascade effect. As for the occurrence of inner filter effect (IFE) and fluorescence resonance energy transfer (FRET), the green fluorescence of N-CDs quenches remarkably, called as OFF initial state. Then the curcumin-F- complex leads to the hypochromatic shift of the absorption band from 532 to 430 nm, which activates the green fluorescence of N-CDs, named as ON state. Meanwhile, the blue fluorescence of N-CDs is quenched due to the FRET, called as OFF terminal state. This system shows good linear relationships from 0 to 35 µM and 0 to 40 µM with low detection limits of 29 nM and 42 nM for curcumin and F- ratiometric detection, respectively. Moreover, a smartphone-assisted analyzer is developed for on-site quantitative detection. Furthermore, we design a logic gate for logistics information storage, which proves the possibility of a logic gate based on N-CDs in practical application. Thus, our work will provide an effective strategy for environmental quantitative monitoring and information storage encryption.

A Novel Nomogram Based on Imaging Biomarkers of Shear Wave Elastography, Angio Planewave Ultrasensitive Imaging, and Conventional Ultrasound for Preoperative Prediction of Malignancy in Patients with Breast Lesions.

Several studies have demonstrated the difficulties in distinguishing malignant lesions of the breast from benign lesions owing to overlapping morphological features on ultrasound. Consequently, we aimed to develop a nomogram based on shear wave elastography (SWE), Angio Planewave Ultrasensitive imaging (Angio PLUS (AP)), and conventional ultrasound imaging biomarkers to predict malignancy in patients with breast lesions. This prospective study included 117 female patients with suspicious lesions of the breast. Features of lesions were extracted from SWE, AP, and conventional ultrasound images. The least absolute shrinkage and selection operator (Lasso) algorithms were used to select breast cancer-related imaging biomarkers, and a nomogram was developed based on six of the 16 imaging biomarkers. This model exhibited good discrimination (area under the receiver operating characteristic curve (AUC): 0.969; 95% confidence interval (CI): 0.928, 0.989) between malignant and benign breast lesions. Moreover, the nomogram also showed demonstrated good calibration and clinical usefulness. In conclusion, our nomogram can be a potentially useful tool for individually-tailored diagnosis of breast tumors in clinical practice.

Energy transfer mediated rapid and visual discrimination of tetracyclines and quercetin in food by using N, Cu Co-doped carbon dots.

The appearance of multi-drug resistant Escherichia coli makes the combination of tetracyclines (TCs) and quercetin (QCT) more common to fight stubborn bacterial infections so that the effective detections of TCs and QCT are essential and necessary. Here, a novel fluorescence probe for differentiating TCs and QCT is developed based on the nitrogen and copper co-doped carbon dots (N, Cu-CDs). The N, Cu-CDs are prepared from ethylene diamine tetraacetic acid (EDTA) and anhydrous copper chloride as precursors through hydrothermal process and exhibit bright blue fluorescence with excellent optical stability. With the presence of four tetracyclines (DOX, TC, CTC and OTC), the fluorescence intensity of N, Cu-CDs is quenched directly due to the internal filtration effect (IFE), and the detection limit obtained through single-signal fluorescence sensing is as low as 23.8 nM for DOX, 37.2 nM for TC, 43.8 nM for OTC and 28.8 nM for CTC. More remarkably, three dimensional ratiometric fluorescence probe for detecting QCT is proposed based on the appearance of another emission at (410 nm, 490 nm) due to electron transform (ET) process. This new method shows a good linear relationship in the range of 10-100 µM with a low detection limit of 59.3 nM. Furthermore, a dual-channel fluorescence sensing platform based on microfluidics paper-based analytical devices (µPADs) is developed for simultaneously visual discrimination of TCs (DOX is chosen as the typical detecting model for TCs) and QCT. This investigation provides a new way for the development of CDs as multifunction fluorescence probes.

Diagnostic Value of Superb Microvascular Imaging in Differentiating Benign and Malignant Breast Tumors: A Systematic Review and Meta-Analysis.

PURPOSE: We performed a systematic review and meta-analysis of studies that investigated the diagnostic performance of Superb Microvascular Imaging (SMI) in differentiating between benign and malignant breast tumors. METHODS: Studies published between January 2010 and March 2022 were retrieved by online literature search conducted in PubMed, Embase, Cochrane Library, Web of Science, China Biology Medicine Disc, China National Knowledge Infrastructure, Wanfang, and Vip databases. Pooled sensitivity, specificity, and diagnostic odd ratios were calculated using Stata software 15.0. Heterogeneity among the included studies was assessed using I2 statistic and Q test. Meta-regression and subgroup analyses were conducted to investigate potential sources of heterogeneity. Influence analysis was conducted to determine the robustness of the pooled conclusions. Deeks' funnel plot asymmetry test was performed to assess publication bias. A summary receiver operating characteristic curve (SROC) was constructed. RESULTS: Twenty-three studies involving 2749 breast lesions were included in our meta-analysis. The pooled sensitivity and specificity were 0.80 (95% confidence interval [CI], 0.77-0.84, inconsistency index [I2] = 28.32%) and 0.84 (95% CI, 0.79-0.88, I2 = 89.36%), respectively. The pooled diagnostic odds ratio was 19.95 (95% CI, 14.84-26.82). The area under the SROC (AUC) was 0.85 (95% CI, 0.81-0.87). CONCLUSION: SMI has a relatively high sensitivity, specificity, and accuracy for differentiating between benign and malignant breast lesions. It represents a promising supplementary technique for the diagnosis of breast neoplasms.

Promising energy transfer system between fuorine and nitrogen Co-doped graphene quantum dots and Rhodamine B for ratiometric and visual detection of doxycycline in food.

A novel sensor based on dual emissive fluorescent graphene quantum dots is developed for a rapid, selective, sensitive and visual detection of doxycycline (DOX). The ratiometric fluorescent probe is designed by grafting fluorescent group (Rhodamine B, RhB) on F, N-doped graphene quantum dots (FNGQDs). In the presence of DOX, the fluorescence at 466 nm is remarkably quenched due to inner filter effect and fluorescence resonance energy transfer, whereas the peak at 592 nm is attenuated slightly due to the energy transfer in the emission peaks of FNGQDs and RhB functional group. The sensor shows good linear relationship from 0.04 to 100 µM with a low detection limit of 40 nM. Furthermore, the flexible solid-state fluorescent sensing platform is used for detecting DOX in milk, pork and water samples. Therefore, this dual-emission FGQD-RhB can be used as a high-performance fluorescent and visual sensor for food safety and environmental monitoring.

A smartphone-integrated optical sensing platform based on Lycium ruthenicum derived carbon dots for real-time detection of Ag.

Growing global environmental pollution problems challenge the need for converting biomass into an advantageous product. In this paper, Lycium ruthenicum is successfully turned into beneficial green emissive (527 nm) fluorescent nitrogen doping carbon dots (N-CDs) via the hydrothermal treatment for the first time. The horizontal and vertical dimensions of N-CDs are demonstrated to be about 4.5 and 0.73 nm, respectively. The N-CDs possess an extremely stable green fluorescence and quantum yield up to 21.8%. Meaningfully, N-CDs exhibit a good linear relationship with Ag+ in the range of 0.7-36 µM, and its detection limit is determined to be 59 nM. The practicability of the fluorescent probe is further validated in lake water and the satisfactory spiked recoveries of Ag+ ranges from 98.99% to 104.19%. Besides, based on the sensitive and selective photoluminescence quenching properties, a smartphone-based laboratory device and RGB analysis software are used to directly capture and analyze fluorescence images with a sensitive detection limit of 83 nM for Ag+. This novel sensor based on N-CDs and smartphone provides a reliable way for on-site monitoring of Ag+ and expands application prospect in the field of environmental pollution detection.

Aldehyde dehydrogenase 1 (ALDH1) immunostaining in axillary lymph node metastases is an independent prognostic factor in ALDH1-positive breast cancer.

OBJECTIVE: To determine whether aldehyde dehydrogenase 1 (ALDH1) immunostaining in axillary lymph node metastases in patients with breast cancer is associated with poor clinical prognosis. METHODS: This retrospective study reviewed data from the medical records of patients with immunohistochemistry-confirmed invasive ductal carcinoma (IDC) and 1-3 metastatic lymph nodes in the ipsilateral axilla between December 2012 and July 2015. The association between ALDH1 immunostaining in axillary lymph node metastases and clinical parameters and prognosis was analysed using χ2-test, Kaplan-Meier survival analysis, univariate and multivariate Cox regression analyses. RESULTS: A total of 229 patients with IDC were enrolled in the study. The median follow-up was 61 months (range, 20-89 months). Patients with ALDH1-positive axillary lymph node metastases had significantly shorter relapse-free survival and overall survival compared with those with ALDH1-negative axillary lymph node metastases. ALDH1 immunostaining in axillary lymph node metastases was a significant predictor of poor prognosis in univariate and multivariate analyses. CONCLUSION: This large study with long-term follow-up suggests that ALDH1 immunostaining in axillary lymph node metastases is an independent predictor of poor prognosis in patients with breast cancer. The clinical relevance of this finding should be confirmed in further well-designed prospective studies.

The nonlinear effect of financial and fiscal policies on poverty alleviation in China-An empirical analysis of Chinese 382 impoverished counties with PSTR models.

Using panel smooth transition regression (PSTR) models, this paper studies the effects of financial and fiscal policies on poverty reduction across 382 poverty-stricken counties in China. The findings are that both fiscal and financial policies have a positive influence on poverty reduction, and their relationships are nonlinear. For either a high or low degree of poverty, fiscal policies are effective for poverty reduction, while financial policies have a greater impact on poverty reduction when there is a medium degree of poverty. Therefore, in deciding which policies should be prioritized for reducing poverty, the level of poverty should be taken into account. To be more specific, when a portfolio of poverty-reduction policies is implemented, fiscal policies should be prioritized at the beginning, when the incidence of poverty is high. Then, financial support should come to the forefront as the poverty level drops, and fiscal support should be stepped up when the poverty level continues to drop.

Optimized Association of Short Alkyl Side Chains Enables Stiff, Self-Recoverable, and Durable Shape-Memory Hydrogel.

This work reports a self-healing and shape-memory hydrogel integrating multiple mechanical properties. The network configuration is featured as entangled networks cross-linked by distributed association of very short alkyl chains (hexyl, six carbons). These cross-linking knots are interconnected by the long hydrophilic polyvinyl alcohol backbone. The optimal aggregation of hexyl side chains leads to the broadened distribution in bonding strength as verified by static and dynamic mechanical characterization. These structural features contribute to high strength, toughness, stiffness, and yet fast recoverability. Furthermore, the hydrophobic and supramolecular nature of aggregated alkyl chains offers high durability and solvent-assistant healing function. Finally, distributed association of hexyl side chains confers a broadened temperature-dependent modulus, allowing for encoding stepwise shape recovery from a temporary shape at different temperatures and/or times.

Facile Soaking Strategy Toward Simultaneously Enhanced Conductivity and Toughness of Self-Healing Composite Hydrogels Through Constructing Multiple Noncovalent Interactions.

Tough and stretchable conductive hydrogels are desirable for the emerging field of wearable and implanted electronics. Unfortunately, most existing conductive hydrogels have low mechanical strength. Current strategies to enhance mechanical properties include employing tough host gel matrices or introducing specific interaction between conductive polymer and host gel matrices. However, these strategies often involve additional complicated processes. Here, a simple yet effective soaking treatment is employed to concurrently enhance mechanical and conductive properties, both of which can be facilely tailored by controlling the soaking duration. The significant improvements are correlated with co-occurring mechanism of deswelling and multiple noncovalent interactions. The resulting optimal sample exhibits attractive combination of high water content (75 wt %), high tensile stress (∼2.5 MPa), large elongation (>600%), reasonable conductivity (∼25 mS/cm), and fast self-healing property with the aid of hot water. The potential application of gel as a strain sensor is demonstrated. The applicability of this method is not limited to conductive hydrogels alone but can also be extended to strengthen other functional hydrogels with weak mechanical properties.

miR-7-5p overexpression suppresses cell proliferation and promotes apoptosis through inhibiting the ability of DNA damage repair of PARP-1 and BRCA1 in TK6 cells exposed to hydroquinone.

Hydroquinone (HQ), one of the major metabolic products of benzene, is a carcinogen, which induces apoptosis and inhibit proliferation in lymphoma cells. microRNA-7-5p (miR-7-5p), a tumor suppressor, participates in various biological processes including cell proliferation and apoptosis regulation by repressing expression of specific oncogenic target genes. To explore whether miR-7-5p is involved in HQ-induced cell proliferation and apoptosis, we assessed the effect of miR-7-5p overexpression on induction of apoptosis analyzed by FACSCalibur flow cytometer in transfection of TK6 cells with miR-7-5p mimic (TK6- miR-7-5p). We observed an increased apoptosis by 25.43% and decreased proliferation by 28.30% in TK6-miR-7-5p cells compared to those negative control cells (TK6-shNC) in response to HQ treatment. Furthermore, HQ might active the apoptotic pathway via partly downregulation the expression of BRCA1 and PARP-1, followed by p53 activation, in TK6-miR-7-5p cells. In contrast, attenuated p53 and BRCA1 expression was observed in shPARP-1 cells than in NC cells after HQ treatment. Therefore, we conclude that HQ may activate apoptotic signals via inhibiting the tumor suppressive effects of miR-7-5p, which may be mediated partly by upregulating the expression of PARP-1 and BRCA1 in control cells. The increase of miR-7-5p expression further intensified downregulation of PARP-1 and BRCA1 in TK6-miR-7-5p cells, resulting in an increase of apoptosis and proliferation inhibited.

Tough and durable hydrogels with robust skin layers formed via soaking treatment.

Recent progress has seen significant advances in the mechanical performances of synthetic hydrogels. However, the mechanics of hydrogels usually suffer drastic degradation under repetitive mechanical and complicated environmental loadings. Here, we fabricate a class of biocompatible layered poly(vinyl alcohol) hydrogels by simply soaking a preformed poly(vinyl alcohol) gel in sodium silicate. The resulting hydrogels exhibit a combination of superior mechanical performances and good biocompatibility, along with exceptional chemical robustness. The layered structure is composed of a compact cover layer and a porous center layer. Both layers are composed of poly(vinyl alcohol). The network in the cover is crosslinked by ordered polarized H-bonds with swelling stability, while the porous center confines a large volume of interstitial water. The structural and crosslinking metric confers the hydrogels with capabilities to tolerate complicated mechanical and environmental loads. The optimized gel is tough (fracture energy >10 kJ m-2) and strong (fracture stress ≈ 5 MPa). The strengthening mechanism can be correlated to the layered structure, which causes the impeded flow of interstitial water. Their mechanical performance is maintained in strong acidic/alkaline, and concentrated electrolytes, and in the presence of salting-in and H-bond-breaking reagents even at elevated temperatures. We speculate that the dipole-dipole pairings of Oδ--Hδ+ Oδ--Hδ+ in the cover layer probably generate hydrophobic microdomains. This swelling-resistant interaction may protect the hydrogels from swelling in complex aqueous envrionments. We also discuss the possible mechanism of the formation of layered structures and their crosslinkings.