Mechanically tough, adhesive, self-healing hydrogel promotes annulus fibrosus repair via autologous cell recruitment and microenvironment regulation.

Annulus fibrosus (AF) defect is an important cause of disc re-herniation after discectomy. The self-regeneration ability of the AF is limited, and AF repair is always hindered by the inflammatory microenvironment after injury. Hydrogels represent one of the most promising materials for AF tissue engineering strategies. However, currently available commercial hydrogels cannot withstand the harsh mechanical load within intervertebral disc. In the present study, an innovative triple cross-linked oxidized hyaluronic acid (OHA)-dopamine (DA)- polyacrylamide (PAM) composite hydrogel, modified with collagen mimetic peptide (CMP) and supplied with transforming growth factor beta 1 (TGF-ß1) (OHA-DA-PAM/CMP/TGF-ß1 hydrogel) was developed for AF regeneration. The hydrogel exhibited robust mechanical strength, strong bioadhesion, and significant self-healing capabilities. Modified with collagen mimetic peptide, the hydrogel exhibited extracellular-matrix-mimicking properties and sustained the AF cell phenotype. The sustained release of TGF-ß1 from the hydrogel was pivotal in recruiting AF cells and promoting extracellular matrix production. Furthermore, the composite hydrogel attenuated LPS-induced inflammatory response and promote ECM synthesis in AF cells via suppressing NFκB/NLRP3 pathway. In vivo, the composite hydrogel successfully sealed AF defects and alleviated intervertebral disk degeneration in a rat tail AF defect model. Histological evaluation showed that the hydrogel integrated well with host tissue and facilitated AF repair. The strategy of recruiting endogenous cells and providing an extracellular-matrix-mimicking and anti-inflammatory microenvironment using the mechanically tough composite OHA-DA-PAM/CMP/TGF-ß1 hydrogel may be applicable for AF defect repair in the clinic. STATEMENT OF SIGNIFICANCE: Annulus fibrosus (AF) repair is challenging due to its limited self-regenerative capacity and post-injury inflammation. In this study, a mechanically tough and highly bioadhesive triple cross-linked composite hydrogel, modified with collagen mimetic peptide (CMP) and supplemented with transforming growth factor beta 1 (TGF-ß1), was developed to facilitate AF regeneration. The sustained release of TGF-ß1 enhanced AF cell recruitment, while both TGF-ß1 and CMP could modulate the microenvironment to promote AF cell proliferation and ECM synthesis. In vivo, this composite hydrogel effectively promoted the AF repair and mitigated the intervertebral disc degeneration. This research indicates the clinical potential of the OHA-DA-PAM/CMP/TGF-ß1 composite hydrogel for repairing AF defects.

Exosomes from M2c macrophages alleviate intervertebral disc degeneration by promoting synthesis of the extracellular matrix via MiR-124/CILP/TGF-ß.

Immuno-inflammation is highly associated with anabolic and catabolic dysregulation of the extracellular matrix (ECM) in the nucleus pulposus (NP), which dramatically propels intervertebral disc degeneration (IVDD). With the characteristics of tissue remodeling and regeneration, M2c macrophages have attracted great attention in research on immune modulation that rebuilds degenerated tissues. Therefore, we first demonstrated the facilitating effects of M2c macrophages on ECM anabolism of the NP in vitro. We subsequently found that exosomes from M2c macrophages (M2c-Exoss) mediated their metabolic rebalancing effects on the ECM. To determine whether M2c-Exoss served as positive agents protecting the ECM in IVDD, we constructed an M2c-Exos-loaded hyaluronic acid hydrogel (M2c-Exos@HA hydrogel) and implanted it into the degenerated caudal disc of rats. The results of MRI and histological staining indicated that the M2c-Exos@HA hydrogel alleviated IVDD in vivo in the long term. To elucidate the underlying molecular mechanism, we performed 4D label-free proteomics to screen dysregulated proteins in NPs treated with M2c-Exoss. Cartilage intermediate layer protein (CILP) was the key protein responsible for the rebalancing effects of M2c-Exoss on ECM metabolism in the NP. With prediction and verification using luciferase assays and rescue experiments, miR-124-3p was identified as the upstream regulator in M2c-Exoss that regulated CILP and consequently enhanced the activity of the TGF-ß/smad3 pathway. In conclusion, we demonstrated ameliorating effects of M2c-Exoss on the imbalance of ECM metabolism in IVDD via the miR-124/CILP/TGF-ß regulatory axis, which provides a promising theoretical basis for the application of M2c macrophages and their exosomes in the treatment of IVDD.

Allicin Alleviates Mitochondrial Dysfunction in Acrylamide-Induced Rat Kidney Involving the Regulation of SIRT1.

Acrylamide (AA), commonly formed in carbohydrate-rich thermally processed foods, exerts harmful effects on the kidney. Allicin, from crushed garlic cloves, exhibits strong biological activities. In the current study, the protection mechanisms of allicin against AA-caused nephrotoxicity were comprehensively examined using an in vivo rat model based on previous research that allicin plays a key role in improving renal function. The results showed that allicin attenuated histological changes of the kidney and ameliorated renal function. Damaged mitochondrial structures, upregulated voltage-dependent anion channel 1 expression, and decreased membrane potential and adenosine 5'-triphosphate levels were observed after AA treatment. Surprisingly, allicin notably reversed the adverse effects. Further, allicin effectively restored mitochondrial function via modulating mitochondrial biogenesis and dynamics, which might be associated with the upregulated expression of sirtuin 1 (SIRT1). Meanwhile, allicin dramatically activated the SIRT1 activity and subsequently inhibited p53 acetylation, prevented the translocation of cytochrome c to the cytoplasm, and reduced the caspase expression, thus further inhibiting mitochondrial apoptosis caused by AA. In summary, the relieving effect of allicin on AA-caused nephrotoxicity lies in its inhibition of mitochondrial dysfunction and mitochondrial apoptosis.

Positive responses and mechanisms of nitrifying sludge to carbon quantum dots: reactor performance, microbial behavior, and antioxidant defense.

Extensive application of carbon quantum dots (CQDs) enlarges its concentration in sewage treatment system. The response of nitrifying sludge to CQDs after long-term exposure was investigated. Results showed that CQD concentrations of 0-100 mg/L presented positive effect to enzymes involved in nitrification, accelerating NH4+-N degradation and NO2--N transformation. The oxidation rate of NO2--N was significantly improved from 3.14 to 7.91 mg/(L h) under the stress of 100 mg/L CQDs. Besides, CQDs stimulated the production of sludge biomass and kept the stability of sludge settleability. Additionally, CQDs were mainly captured by loosely bound extracellular polymeric substances, reducing aromatic-like protein. Microbes alleviated CQD stress by secreting tryptophan-like protein and polysaccharides. After few CQDs entered cells, intracellular antioxidant defense was activated. Total antioxidant capacity level was heightened at least 31%. The activities of superoxide dismutase and catalase were enhanced at relatively low and high CQD concentration levels. Hence, microbial metabolic pathways, microbial community, and nitrifying bacteria were not significantly affected by CQDs. The findings of this work provide new insight for understanding the environmental implication of CQDs in the biological treatment system.

Plant base editing and prime editing: The current status and future perspectives.

Precise replacement of an allele with an elite allele controlling an important agronomic trait in a predefined manner by gene editing technologies is highly desirable in crop improvement. Base editing and prime editing are two newly developed precision gene editing systems which can introduce the substitution of a single base and install the desired short indels to the target loci in the absence of double-strand breaks and donor repair templates, respectively. Since their discoveries, various strategies have been attempted to optimize both base editor (BE) and prime editor (PE) in order to improve the precise editing efficacy, specificity, and expand the targeting scopes. Here, we summarize the latest development of various BEs and PEs, as well as their applications in plants. Based on these progresses, we recommend the appropriate BEs and PEs for both basic plant research and crop improvement. Moreover, we propose the perspectives for further optimization of these two editors. We envision that both BEs and PEs will become the routine and customized precise gene editing tools for both plant biological research and crop improvement in the near future.

Clinical application of bronchial high-frequency ventilation in 2-port thoracoscopic segmentectomy.

To evaluate the safety and clinical application of a computer-aided surgery system (CAS) combined with high-frequency bronchial ventilation in 2-port thoracoscopic anatomical segmentectomy. A total of 301 patients who underwent 2-port thoracoscopic segmentectomy between January 1, 2019 and March 1, 2022 in the 960th Hospital of the People's Liberation Army and the Department of Thoracic Surgery of Zibo Municipal Hospital were retrospectively analyzed. The experimental and control groups were created according to the different methods of appearing the intersegmental plane of the lung. The experimental group comprised 152 patients who underwent CAS reconstruction combined with high-frequency ventilation, and the control group comprised 149 patients who underwent CAS reconstruction combined with expansion collapse. The characteristics of the patients, including age, sex, smoking history, forced expiratory volume in 1 second/forced vital capacity, Maximal ventilation, diameter of pulmonary nodules, intraoperative blood loss, postoperative drainage volume, drainage tube removal time, length of hospital stay after extubation, postoperative complication rate, operation time and appearance time of the intersegmental plane, were compared between the 2 groups. All patients completed the operation between high-frequency bronchial ventilation and expansion collapse group. There was no significant difference in Forced expiratory volume in 1 second/Forced vital capacity [(101.05 ±â€…11.86) vs (101.86 ±â€…11.61)], maximum expiratory volume [(86.36 ±â€…17.59 L) vs (85.28 ±â€…17.68 L)], the diameter of lung nodules [(13.61 ±â€…3.51 cm) vs (13.21 ±â€…3.41 cm)], intraoperative blood loss [(47.50 ±â€…45.90 mL) vs (48.49 ±â€…34.65 mL)], postoperative drainage volume [(425.16 ±â€…221.61 mL) vs (444.70 ±â€…243.72 mL)], drainage tube removal time [(3.88 ±â€…1.85 days) vs (3.43 ±â€…1.81 days)], or postoperative hospital stay [(6.07 ±â€…2.14 days) vs (5.82 ±â€…1.88 days) between the experimental group and the control group (P > .05)]. There were significant differences in operation time [(95.05 ±â€…26.85 min) vs (117.85 ±â€…31.70 min), P = .017] and intersegmental plane appearance time [(2.37 ±â€…1.03 min) vs (14.20 ±â€…3.23 min), P < .001]. High-frequency bronchial ventilation is safe and feasible when used in quickly and accurately identifying the intersegmental plane and is worthy of clinical application in 2-port thoracoscopic segmentectomy.

Modulation of Disordered Bile Acid Homeostasis and Hepatic Tight Junctions Using Salidroside against Hepatocyte Apoptosis in Furan-Induced Mice.

Furan, a processing-induced food contaminant, has attracted great attention due to its hepatotoxicity. To further investigate the underlying mechanism of salidroside (SAL) alleviating furan-induced liver damage, we divided Balb/c mice into the control group, the furan (8 mg/kg/day) group, and three groups of three different doses of SAL (10/20/40 mg/kg/day) in the current research. The shifted serum profile was observed through untargeted metabonomics, to which the bile acid metabolism was related, and the alleviating effect of SAL against furan-induced apoptosis was caused by the metabolism. Target bile acid quantification for the liver and serum showed that SAL positively regulated the homeostasis of bile acids disturbed by furan. Meanwhile, SAL significantly upregulated the synthesis genes of bile acids (Cyp7a1, Cyp7b1, Cyp8b1, and Cyp27a1) and the uptake transport genes (Ntcp and Oatps) and downregulated the efflux transport genes (Bsep, Ost-α, Ost-ß, Mrp2, and Mrp4). Transmission electron microscopy of the bile canaliculi and tight junctions and the levels of tight junction marker proteins (ZO-1, occludin, and claudin-1) confirmed that the disruption of the hepatic tight junction was inhibited by SAL. The connection between the apoptosis- and tight junction-related proteins was observed through the construction of a protein-protein interaction network. SAL suppressed the furan-induced hepatocyte apoptosis evidenced by the detection of TUNEL and Bax, Bcl-2, and caspase-3 levels. Taken together, SAL alleviated furan-induced hepatocyte apoptosis via altering the disordered homeostasis of bile acids and hepatic tight junctions.

Perioperative Acupuncture Optimizes Surgical Outcomes: Theory, Clinical Practice and Future Perspectives.

A growing body of evidence supports the use of perioperative acupuncture as part of an enhanced postsurgical recovery protocol. Data from both clinical trials and animal studies has shown that the integration of acupuncture into perioperative patient care leads to a reduction of perioperative complications such as preoperative anxiety, intraoperative hemodynamic instability, postoperative pain, postoperative cognitive dysfunction, and postoperative nausea and vomiting in surgical patients. Despite these favorable outcomes, perioperative acupuncture has yet to be widely adopted in current anesthesia practice. This review summarized data from clinical perioperative acupuncture studies and cites recent discoveries regarding the anatomical location and characteristics of acupoint(s), acupuncture stimulation techniques, and treatment practice protocols, as well as identified the areas of deficiency in perioperative acupuncture applications. To facilitate acupuncture integration in perioperative care practice, the authors propose to establish a perioperative acupuncture registry which can be used for data mining as well as a resource for studying the underlying mechanisms of acupuncture. Through this acupuncture registry, clinical guidelines and research protocols can be established, additional large/multi-center clinical and pragmatic trials can be easily performed to determine if the integration and expansion of perioperative acupuncture practice is cost-effective.

A donor-DNA-free CRISPR/Cas-based approach to gene knock-up in rice.

Structural variations (SVs), such as inversion and duplication, contribute to important agronomic traits in crops1. Pan-genome studies revealed that SVs were a crucial and ubiquitous force driving genetic diversification2-4. Although genome editing can effectively create SVs in plants and animals5-8, the potential of designed SVs in breeding has been overlooked. Here, we show that new genes and traits can be created in rice by designed large-scale genomic inversion or duplication using CRISPR/Cas9. A 911 kb inversion on chromosome 1 resulted in a designed promoter swap between CP12 and PPO1, and a 338 kb duplication between HPPD and Ubiquitin2 on chromosome 2 created a novel gene cassette at the joint, promoterUbiquitin2::HPPD. Since the original CP12 and Ubiquitin2 genes were highly expressed in leaves, the expression of PPO1 and HPPD in edited plants with homozygous SV alleles was increased by tens of folds and conferred sufficient herbicide resistance in field trials without adverse effects on other important agronomic traits. CRISPR/Cas-based genome editing for gene knock-ups has been generally considered very difficult without inserting donor DNA as regulatory elements. Our study challenges this notion by providing a donor-DNA-free strategy, thus greatly expanding the utility of CRISPR/Cas in plant and animal improvements.

A Novel Feature Extraction Method for Power Transformer Vibration Signal Based on CEEMDAN and Multi-Scale Dispersion Entropy.

Effective diagnosis of vibration fault is of practical significance to ensure the safe and stable operation of power transformers. Aiming at the traditional problems of transformer vibration fault diagnosis, a novel feature extraction method based on complete ensemble empirical mode decomposition with adaptive noise (CEEMDAN) and multi-scale dispersion entropy (MDE) was proposed. In this paper, CEEMDAN method is used to decompose the original transformer vibration signal. Additionally, then MDE is used to capture multi-scale fault features in the decomposed intrinsic mode functions (IMFs). Next, the principal component analysis (PCA) method is employed to reduce the feature dimension and extract the effective information in vibration signals. Finally, the simplified features are sent into density peak clustering (DPC) to get the fault diagnosis results. The experimental data analysis shows that CEEMDAN-MDE can effectively extract the information of the original vibration signals and DPC can accurately diagnose the types of transformer faults. By comparing different algorithms, the practicability and superiority of this proposed method are verified.

A Novel Hybrid Approach for Partial Discharge Signal Detection Based on Complete Ensemble Empirical Mode Decomposition with Adaptive Noise and Approximate Entropy.

To eliminate the influence of white noise in partial discharge (PD) detection, we propose a novel method based on complete ensemble empirical mode decomposition with adaptive noise (CEEMDAN) and approximate entropy (ApEn). By introducing adaptive noise into the decomposition process, CEEMDAN can effectively separate the original signal into different intrinsic mode functions (IMFs) with distinctive frequency scales. Afterward, the approximate entropy value of each IMF is calculated to eliminate noisy IMFs. Then, correlation coefficient analysis is employed to select useful IMFs that represent dominant PD features. Finally, real IMFs are extracted for PD signal reconstruction. On the basis of EEMD, CEEMDAN can further improve reconstruction accuracy and reduce iteration numbers to solve mode mixing problems. The results on both simulated and on-site PD signals show that the proposed method can be effectively employed for noise suppression and successfully extract PD pulses. The fusion algorithm combines the CEEMDAN algorithm and the ApEn algorithm with their respective advantages and has a better de-noising effect than EMD and EEMD.

Cancer inhibition mechanism of lung cancer mouse model based on dye trace method.

To minimize the incidence and mortality of cancer, dye trace method was used to explore the mechanism of drug inhibition. 60 mice were selected as the research objects and randomly divided into five groups: model group, shikonin group, aconitine group, notoginsenoside R1 group, and compound group. When establishing the model, begin to administrate the medicine by gavage. The permeability of lung barrier was measured, and H.E staining, immunohistochemical staining, and Western blot test were carried out. The results showed that the mice in model group had decreased autonomic activity, increased permeability of the lung barrier, white nodules on the lung tissue, decreased protein expression related to cell proliferation and differentiation, and decreased protein expression associated with cell proliferation and differentiation, increased expression of related proteins in cancer stem cells, and low level of cell-linked communication. And the incidence of lung cancer in the model group mice was 100%. The histopathological changes in mice were improved to varying degrees after the intervention of the three drugs. Especially in the compound group, the incidence of lung cancer decreased to 8.3%. This study demonstrated that the combination of shikonin, aconitine and notoginsenoside R1 had a good anti-cancer effect, which provided a theoretical basis for clinical research.

From two- to multi-state vertical spin valves without spacer layer based on Fe3GeTe2 van der Waals homo-junctions.

Different than covalently bonded magnetic multilayer systems, high-quality interfaces without dangling bonds in van der Waals (vdW) junctions of two-dimensional (2D) layered magnetic materials offer opportunities to realize novel functionalities. Here, we report the fabrication of multi-state vertical spin valves without spacer layers by using vdW homo-junctions in which exfoliated Fe3GeTe2 nanoflakes act as ferromagnetic electrodes and/or interlayers. We demonstrate the typical behavior of two-state and three-state magnetoresistance for devices with two and three Fe3GeTe2 nanoflakes, respectively. Distinct from traditional spin valves with sandwich structures, our novel homo-junction-based spin-valve structure allows the straightforward realization of multi-state magnetic devices. Our work demonstrates the possibility of extend multi-state, non-volatile spin information to 2D magnetic homo-junctions, and it emphasizes the utility of vdW interface as a fundamental building block for spintronic devices.

A comprehensive comparison of posterior lumbar interbody fusion versus posterolateral fusion for the treatment of isthmic and degenerative spondylolisthesis: A meta-analysis of prospective studies.

Posterolateral fusion (PLF) and posterior lumbar interbody fusion (PLIF) are both frequently-used procedures for the surgical treatment of isthmic and degenerative spondylolisthesis. The current meta-analysis aimed to perform a comprehensive comparison between PLF and PLIF in terms of the therapeutic effects on spondylolisthesis. PubMed, Embase, Web of Science and the Cochrane Central Register of Controlled Trials were searched for relevant prospective studies. Measures of clinical outcome, postoperative complication rate, fusion rate, and blood loss are presented as odds ratio (OR), mean difference and corresponding 95 % confidence interval (CI) as appropriate. Eight prospective studies comprising 723 patients were eventually enrolled in the meta-analysis. Patients who underwent PLIF had a better clinical outcome (pooled OR, 1.63 [95 % CI, 1.02-2.61]; p = 0.04) and a higher fusion rate (pooled OR, 3.33 [95 % CI, 1.88-5.90]; p < 0.01) than those who underwent PLF. No significant difference between the two procedures was identified for postoperative complication rate and blood loss. The results showed that PLIF was superior to PLF in clinical outcome and fusion rate, and equal to PLF in terms of blood loss and the rate of postoperative complications. Here we provide the most effective evidence currently available for the comparison between PLF and PLIF, which has guiding significance for clinical practice.

Seizures at presentation are correlated with better survival outcomes in adult diffuse glioma: A systematic review and meta-analysis.

PURPOSE: Seizures are the most common presenting sign of patients with diffuse glioma. In the current study, we performed a meta-analysis to determine the correlation of seizures at presentation to survival outcomes in adult diffuse glioma, and the possible mechanisms were also discussed. METHODS: A comprehensive literature search was performed in PUBMED, EMBASE, Web of Science and the Cochrane Central Register of Controlled Trials. The pooled hazard ratio (HR) and corresponding 95% confidence interval (CI) were used to estimate effects. Heterogeneity among studies and publication bias were also evaluated. RESULTS: 11 studies with 2088 patients were finally included for the current meta-analysis. Seizure-free preoperatively was significantly associated with a poor overall survival in patients with diffuse glioma, the pooled HR was 1.73 (95% CI 1.43-2.08, Z = 5.71, p < 0.001). Subgroup analysis was also performed by tumor grade, the same association was identified in both low-grade glioma (pooled HR 2.49, 95% CI 1.47-4.20, Z = 3.40, p < 0.001) and glioblastoma (pooled HR 1.46, 95% CI 1.27-1.68, Z = 5.24, p < 0.001). A significant correlation of seizure-free with a poor progression-free survival was also identified (pooled HR 1.42, 95% CI 1.06-1.92, Z = 2.33, p = 0.02), although only 3 studies comprising 368 patients were included. CONCLUSION: The current study determined that seizures at presentation were an independent predictor of better survival outcomes in adult diffuse glioma. It is the first study which provides a comprehensive standardized assessment of the association between seizures at presentation with long-term survival outcomes in patients with diffuse glioma.

IDH1 mutation is associated with a higher preoperative seizure incidence in low-grade glioma: A systematic review and meta-analysis.

PURPOSE: Gliomas, particularly low-grade gliomas (LGGs), are highly epileptogenic. Seizure is the most common presenting sign of LGG patients and significantly decreases their quality of life. Accordingly, there is a need for a better understanding of the mechanisms and risk factors of glioma-related epilepsy. The current study aimed to perform a comprehensive meta-analysis to investigate the correlation of isocitrate-dehydrogenase 1 (IDH1), an important molecular biomarker for glioma classification and prognosis, to preoperative seizure incidence in LGG. METHODS: PUBMED, EMBASE, and Web of Science databases were searched for relevant studies. The odds ratio (OR) and corresponding 95% confidence interval (CI) were used as the primary measures to assess the correlation between IDH1 mutation and preoperative seizure incidence. RESULTS: A total of 722 LGG patients, including 555 patients with IDH1 mutation and 167 patients with wild-type IDH1 were enrolled in the current meta-analysis. The pooled OR was 2.47 (95% CI 1.70-3.57, Z = 4.78, p < 0.01). No significant heterogeneity was observed among all included studies and no publication bias was identified. CONCLUSION: The current meta-analysis identified that IDH1 mutation was correlated to a higher preoperative seizure incidence in LGG. This result would generate impetus for research on the mechanisms behind this correlation, and provide a new idea for the individualized treatment of glioma-related epilepsy.

Application of shielding boronate affinity chromatography in the study of the glycation pattern of haemoglobin.

Human haemoglobin (Hb) may appear in a number of glycated species. The glycation pattern of Hb using shielding boronate affinity chromatography (SBAC) has been studied in the present work. SBAC is a novel separation technique, which eliminates nonspecific boronate-protein interactions by introducing a so-called shielding reagent. Two samples from Bio-Rad (Lyphochek)--one from normal persons' blood with relatively low HbA(1c) level (HbL) and the other from diabetic patients' blood with an elevated HbA(1c) level (HbH)--were used for the investigation. Glycated Hb (GHb) was separated from nonglycated Hb species using Tris as the shielding reagent. Two eluted peaks, eluted peak 1 (E1) and eluted peak 2 (E2), were obtained using a linear gradient elution with Tris. Several bands were observed on isoelectric focusing gel, which showed the same migration positions as Hb adducts, such as HbA(0), which is major Hb component containing two alpha chains and two beta chains; HbA(1c), which is post-translational glycation on the N-terminus of the beta chains of HbA(0); Foetal Hb (HbF), consisting of two alpha chains and two gamma chains; and glutathione Hb (also called HbSSG), which is the result from thiol-disulphide interchain exchange during oxidation of the thiol groups of Hb. In both HbL and HbH samples, E2 exhibited slightly higher amounts of HbF than E1. Electrospray-ionisation mass spectrometry showed that: (1) HbL-E1 was glycated with single glucose on both alpha and beta chains while no observable glycated chains were present in HbL-E2; (2) both HbH-E1 and HbH-E2 were glycated with single glucoses on both alpha and beta chains, however, compared with HbH-E1, HbH-E2 showed a higher relative intensity of the glycated beta chain and lower relative intensity of the glycated alpha chain; and (3) the degree of glycation increased with increasing glycation level of the sample. The amount of HbA(1c) presented in the eluted peaks was further determined using enzymatic digestion of glycated Hb by endoproteinase Glu-C and the subsequent separation and analysis of the digested peptides by reversed-phase high-performance liquid chromatography and capillary electrophoresis. The values of HbA(1c)/HbA(0) of the eluted peaks, i.e. HbL-E1, HbL-E2, HbH-E1 and HbH-E2, were 0.27, 0.19, 0.50 and 0.43, respectively. In both HbL and HbH samples, E1 contained higher amounts of HbA(1c) than E2. This study demonstrates the structural heterogeneity of GHb as well as the possibility of using SBAC to detect glycated species of Hb.