Polyphenol (-)-Epigallocatechin Gallate (EGCG) mitigated kidney injury by regulating metabolic homeostasis and mitochondrial dynamics involvement with Drp1-mediated mitochondrial fission in mice.

The study aimed to examine effects of (-)-epigallocatechin-3-gallate (EGCG) on energy metabolism and mitochondrial dynamics in mouse model of renal injury caused by doxorubicin (DOX). Here, mice were divided into Control group, EGCG-only treated group, DOX group, and three doses of EGCG plus DOX groups. Our results showed that EGCG behaved beneficial effects against kidney injury via attenuation of pathological changes in kidney tissue, which was confirmed by reducing serum creatinine (SCr), blood urea nitrogen (BUN), and apoptosis. Subsequently, changes in reactive oxygen species generation, malondialdehyde content, and activities of antioxidant enzymes were considerably ameliorated in EGCG + DOX groups when compared to DOX group. Furthermore, EGCG-evoked renal protection was associated with increases of mitochondrial membrane potential and decreases of mitochondrial fission protein Dynamin-related protein 1 (Drp1). Moreover, changing glycolysis into mitochondrial oxidative phosphorylation was observed, evidenced by controlling activities of malate dehydrogenase (MDH) and hexokinase (HK) in EGCG + DOX groups when compared to DOX group, indicating that reprogramming energy metabolism was linked to EGCG-induced renal protection in mice. Therefore, EGCG was demonstrated to have a protective effect against kidney injury by reducing oxidative damage, metabolic disorders, and mitochondrial dysfunction, suggesting that EGCG has potential as a feasible strategy to prevent kidney injury.

miR-455/GREM1 axis promotes colorectal cancer progression and liver metastasis by affecting PI3K/AKT pathway and inducing M2 macrophage polarization.

BACKGROUND: Colorectal cancer is among the most common malignant tumors affecting the gastrointestinal tract. Liver metastases, a complication present in approximately 50% of colorectal cancer patients, are a considerable concern. Recently, studies have revealed the crucial role of miR-455 in tumor pathogenesis. However, the effect of miR-455 on the progression of liver metastases in colorectal cancer remains controversial. As an antagonist of bone morphogenetic protein(BMP), Gremlin 1 (GREM1) may impact organogenesis, body patterning, and tissue differentiation. Nevertheless, the role of miR-455 in regulating GREM1 in colorectal cancer liver metastases and how miR-455/GREM1 axis influences tumour immune microenvironment is unclear. METHODS: Bioinformatics analysis shows that miR-455/GREM1 axis plays crucial role in liver metastasis of intestinal cancer and predicts its possible mechanism. To investigate the impact of miR-455/GREM1 axis on the proliferation, invasion, and migration of colorectal cancer cells, colony formation assay, wound healing and transwell assay were examined in vitro. The Dual-Luciferase reporter gene assay and RNA pull-down assay confirmed a possible regulatory effect between miR-455 and GREM1. In vivo, colorectal cancer liver metastasis(CRLM) model mice was established to inquiry the effect of miR-455/GREM1 axis on tumor growth and macrophage polarization. The marker of macrophage polarization was tested using immunofluorescence(IF) and quantitative real-time polymerase chain reaction(qRT-PCR). By enzyme-linked immunosorbent assay (ELISA), cytokines were detected in culture medium supernatants. RESULTS: We found that miR-455 and BMP6 expression was increased and GREM1 expression was decreased in liver metastase compared with primary tumor. miR-455/GREM1 axis promotes colorectal cancer cells proliferation, migration, invasion via affected PI3K/AKT pathway. Moreover, downregulating GREM1 augmented BMP6 expression in MC38 cell lines, inducing M2 polarization of macrophages, and promoting liver metastasis growth in CRLM model mice. CONCLUSION: These data suggest that miR-455/GREM1 axis promotes colorectal cancer progression and liver metastasis by affecting PI3K/AKT pathway and inducing M2 macrophage polarization. These results offer valuable insights and direction for future research and treatment of CRLM.

A Fluorescent In Situ Hybridization (FISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Cell Culture.

Hepatitis B virus (HBV) is undoubtedly a master in exploiting host resources while evading host defense for its multiplication within a constrained genetic coding capacity. To further unravel these cunning strategies, a clear picture of virus-host interaction with key subcellular and molecular contexts is needed. Here, we describe a FISH protocol modified from the ViewRNA assay that allows direct visualization of HBV RNA, DNA, and cccDNA in cell culture models (e.g., HepAD38, HepG2-NTCP). It can be coupled with immunofluorescence staining of viral or host proteins or other fluorescent tagging systems which could illuminate numerous aspects of virus-host interactions.

Comparison of biomechanical effects of polyetheretherketone (PEEK) rods and titanium rods in lumbar long-segment instrumentation: a finite element study.

Introduction: Polyetheretherketone (PEEK) lumbar fusion rods have been successfully used in short-segment posterior instrumentation to prevent adjacent segment degeneration. However, limited studies have reported their application in lumbar long-segment instrumentation. This study aimed to compare the biomechanical performances of PEEK rods and titanium rods in lumbar long-segment instrumentation using finite element (FE) models, with the expectation of providing clinical guidance. Methods: A lumbar FE model (A) and four lumbar fixation FE models (BI, CI, BII, CII) of the L1-S1 vertebral body were developed using CT image segmentation (A: intact model; BI: intact model with L2-S1 PEEK rod internal fixation; CI: intact model with L2-S1 titanium rod internal fixation; BII: intact model with L3-S1 PEEK rod internal fixation; CII: intact model with L3-S1 titanium rod internal fixation). A 150-N preload was applied to the top surface of L1, similar to the intact model. The stresses on the lumbar intervertebral disc, facet joint, pedicle screws, and rods were calculated to evaluate the biomechanical effect of the different fixation procedures in lumbar long-segment instrumented surgery. Results: Under the four physiological motion states, the average stresses on the adjacent segment intervertebral disc and facet joint in all fixation models were greater than those in the intact model. Furthermore, the average stresses on the adjacent segment intervertebral disc and facet joint were greater in models CI and CII than in models BI and BII, respectively. The average stresses on the pedicle screws and rods were decreased in models BI and BII compared with models CI and CII under the four physiological motion states, respectively. Discussion: The PEEK rod internal fixation system may have better biomechanical properties than the titanium rod internal fixation system in delaying adjacent segment degeneration, improving the lumbar function of postoperative patients, and reducing the risk of screw loosening and breakage in lumbar long-segment instrumentation.

Necroptosis in bacterial infections.

Necroptosis, a recently discovered form of cell-programmed death that is distinct from apoptosis, has been confirmed to play a significant role in the pathogenesis of bacterial infections in various animal models. Necroptosis is advantageous to the host, but in some cases, it can be detrimental. To understand the impact of necroptosis on the pathogenesis of bacterial infections, we described the roles and molecular mechanisms of necroptosis caused by different bacterial infections in this review.

Taxonomy and phylogeny of Panus (Polyporales, Panaceae) in China and its relationship with allies.

Panus is a typical wood-rotting fungi, which plays considerable roles in ecosystems and has significant economic value. The genus Panus currently consists of more than 100 species; however, only eight species have been reported from China. This study aims to distinguish and describe two novel species from the Panussimilis complex, namely Panusminisporus and Panusbaishanzuensis, one new record species from Zhejiang Province, Panussimilis and three common species, Panusconchatus, Panusneostrigosus and Panusrudis, based on detailed morphological and phylogenetic studies, relying on Chinese specimens. Panusminisporus is characterised by its reddish-brown pileus, decurrent lamellae with cross-veins, slender stipe, smaller basidiospores, wider generative hyphae and absence of sclerocystidia. Panusbaishanzuensis is featured by its pileus with concentric and darker ring zone, decurrent lamellae with cross-veins, shorter stipe, longer basidiospores, diverse and shorter cheilocystidia and smaller sclerocystidia. Internal transcribed spacer (ITS) regions, large subunit nuclear ribosomal RNA gene (nLSU) and translation elongation factor 1-α gene (tef-1α) were employed to perform a thorough phylogenetic analysis for genus Panus and related genera, using Bayesian Inference and Maximum Likelihood analysis. The results indicate that Panusminisporus and Panusbaishanzuensis form two independent clades within the Panussimilis complex themselves. Detailed descriptions, taxonomic notes, illustrations etc. were provided. In addition, a key to the reported species of Panus from China is also provided.

Tumor-resident microbiota contributes to colorectal cancer liver metastasis by lactylation and immune modulation.

The role of tumor-resident microbiota in modulating tumor immunity remains unclear. Here, we discovered an abundance of intra-tumoral bacteria, such us E.coli, residing and resulting in Colorectal cancer liver metastasis (CRLM). E.coli enhanced lactate production, which mediated M2 macrophage polarization by suppressing nuclear factor-κB -gene binding (NF-κB) signaling through retinoic acid-inducible gene 1 (RIG-I) lactylation. Lactylation of RIG-I suppressed recruitment of NF-κB to the Nlrp3 promoter in macrophages, thereby reducing its transcription. This loss of Nlrp3 affected the immunosuppressive activities of regulatory T cells (Tregs) and the antitumor activities of and CD8+ T cells. Small-molecule compound screening identified a RIG-I lactylation inhibitor that suppressed M2 polarization and sensitized CRLM to 5-fluorouracil (5-FU). Our findings suggest that tumor-resident microbiota may be a potential target for preventing and treating CRLM.

Quantitative analysis from ultrafast dynamic contrast-enhanced breast MRI using population-based versus individual arterial input functions, and comparison with semi-quantitative analysis.

PURPOSE: To evaluate the value of inline quantitative analysis of ultrafast dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) using a population-based arterial input function (P-AIF) compared with offline quantitative analysis with an individual AIF (I-AIF) and semi-quantitative analysis for diagnosing breast cancer. METHODS: This prospective study included 99 consecutive patients with 109 lesions (85 malignant and 24 benign). Model-based parameters (Ktrans, kep, and ve) and model-free parameters (washin and washout) were derived from CAIPIRINHA-Dixon-TWIST-VIBE (CDTV) DCE-MRI. Univariate analysis and multivariate logistic regression analysis with forward stepwise covariate selection were performed to identify significant variables. The AUC and F1 score were assessed for semi-quantitative and two quantitative analyses. RESULTS: kep from inline quantitative analysis with P-AIF for diagnosing breast cancer provided an AUC similar to kep from offline quantitative analysis with I-AIF (0.782 vs 0.779, p = 0.954), higher compared to washin from semi-quantitative analysis (0.782 vs 0.630, p = 0.034). Furthermore, the inline quantitative analysis with P-AIF achieved the larger F1 score (0.920) compared with offline quantitative analysis with I-AIF (0.780) and semi-quantitative analysis (0.480). There were no statistically significant differences for kep values between the two quantitative analysis schemes (p = 0.944). CONCLUSION: The inline quantitative analysis with P-AIF from CDTV in characterizing breast lesions could offer similar diagnostic accuracy to offline quantitative analysis with I-AIF, and higher diagnostic accuracy to semi-quantitative analysis.

Chinese Medicine Prolongs Overall Survival of Chinese Patients with Advanced Gastric Cancer: Treatment Pattern and Survival Analysis of a 20-Year Real-World Study.

OBJECTIVE: To describe the treatment patterns and survival status of advanced gastric cancer (AGC) in China in the past two decades, and objectively evaluate the impact of standardized Chinese medicine (CM) treatment on the survival of AGC patients. METHODS: This multicenter registry designed and propensity score analysis study described the diagnosis characteristics, treatment-pattern development and survival status of AGC from 10 hospitals in China between January 1, 2000 and July 31, 2021. Overall survival (OS) was evaluated between non-CM cohort (standard medical treatment) and CM cohort (integrated standard CM treatment ≥3 months). Propensity score matching (PSM) and inverse probability of treatment weighting (IPTW) were performed to adjust any difference in average outcomes for bias. RESULTS: A total of 2,001 patients histologically confirmed locally advanced and/or metastasis stomach and gastroesophageal junction adenocarcinoma were enrolled. Among them, 1,607 received systemic chemotherapy, 215 (10.74%) accepted molecular targeted therapy, 44 (2.2%) received checkpoint inhibitor therapy, and 769 (38.43%) received CM. Two-drug regimen was the main choice for first-line treatment, with fluoropyrimidine plus platinum as the most common regimen (530 cases, 60.09%). While 45.71% (16 cases) of patients with HER2 amplification received trastuzumab in first-line. The application of apatinib increased (33.33%) in third-line. The application of checkpoint inhibitors has increased since 2020. COX analysis showed that Lauren mixed type (P=0.017), cycles of first-line treatment >6 (P=0.000), CM (P=0.000), palliative gastrectomy (P=0.000), trastuzumab (P=0.011), and apatinib (P=0.008) were independent prognostic factors for the OS of AGC. After PSM and IPTW, the median OS of CM cohort and non-CM cohort was 18.17 and 12.45 months, respectively (P<0.001). CONCLUSIONS: In real-world practice for AGC in China, therapy choices consisted with guidelines. Two-drug regimen was the main first-line choice. Standardized CM treatment was an independent prognostic factor and could prolong the OS of Chinese patients with AGC. (Registration No. NCT02781285).

Ventricular Netrin-1 deficiency leads to defective pyramidal decussation and mirror movement in mice.

The corticospinal tract (CST) is the principal neural pathway responsible for conducting voluntary movement in the vertebrate nervous system. Netrin-1 is a well-known guidance molecule for midline crossing of commissural axons during embryonic development. Families with inherited Netrin-1 mutations display congenital mirror movements (CMM), which are associated with malformations of pyramidal decussation in most cases. Here, we investigated the role of Netrin-1 in CST formation by generating conditional knockout (CKO) mice using a Gfap-driven Cre line. A large proportion of CST axons spread laterally in the ventral medulla oblongata, failed to decussate and descended in the ipsilateral spinal white matter of Ntn1Gfap CKO mice. Netrin-1 mRNA was expressed in the ventral ventricular zone (VZ) and midline, while Netrin-1 protein was transported by radial glial cells to the ventral medulla, through which CST axons pass. The level of transported Netrin-1 protein was significantly reduced in Ntn1Gfap CKO mice. In addition, Ntn1Gfap CKO mice displayed increased symmetric movements. Our findings indicate that VZ-derived Netrin-1 deletion leads to an abnormal trajectory of the CST in the spinal cord due to the failure of CST midline crossing and provides novel evidence supporting the idea that the Netrin-1 signalling pathway is involved in the pathogenesis of CMM.

New species, new records and common species of Pluteussect.Celluloderma from northern China.

Wood-rotting fungi are organisms that can decompose wood substrates and extract nutrients from them to support their growth. They play a crucial role in the material cycle of forest ecosystems. The genus Pluteus plays a significant role in wood decomposition. In this study, the morphology and molecular systematics of the sect. Celluloderma of the genus Pluteus were carried out. Pluteusbrunneodiscus was identified as a new species, along with the discovery of two new records, P.cystidiosus and P.chrysophlebius, and a common species, P.romellii. Pluteusbrunneodiscus is characterized by the brown center of the pileus that transitions to white towards the margins, with the surface cracking to form irregular granules. It is typically found in Populus forests growing on decomposing twigs or wood chips. Line drawings, color photographs, and phylogenetic analyses of related species within the genus Pluteus accompany the descriptions of these four species. The analyses are based on ITS + TEF1-α sequence data. Finally, a key for the twenty species within the sect. Celluloderma of the genus Pluteus, which has been documented in China, is provided.

Ortho C-H Bond Activations in an Atmospheric Microwave Plasma Ion Source.

C-H bond ortho-substitution reaction has always been a significant and challenging topic in organic chemistry. We proposed a synthesis method based on microwave plasma torches. High-resolution mass spectrometry was used to monitor rapid reaction products. 2-Alkylbenzimidazole can be formed through the reaction of phenylnitrenium ion and nitriles on a millisecond scale. This reaction can achieve the one-step formation of benzimidazoles from benzene ring single-substituted compounds without the addition of external oxidants or catalysts. A similar C-H bond activation reaction can be accomplished with ketones. Meanwhile, the microwave plasma reactor was modified, and the resulting 2-methylbenzimidazole was successfully collected, indicating the device has good application potential in organic reactions such as C-H bond activation reaction.

Multiple evidence reveals two new species and new distributions of Calocybe species (Lyophyllaceae) from northeastern China.

The Calocybe species possess notable economic and medicinal value, demonstrating substantial potential for resource utilization. The taxonomic studies of Calocybe are lacking in quality and depth. Based on the specimens collected from northeast China, this study provides a detailed description of two newly discovered species, namely Calocybebetulicola and Calocybecystidiosa, as well as two commonly found species, Calocybedecolorata and Calocybeionides. Additionally, a previously unrecorded species, C.decolorata, has recently been discovered in Jilin Province, China. The two newly discovered species can be accurately distinguished from other species within the genus Calocybe based on their distinct morphological characteristics. The primary distinguishing features of C.betulicola include its grayish-purple pileus, grayish-brown to dark purple stipe, smaller basidiomata, absence of cellular pileipellis, and its habitat on leaf litter within birch forests. Calocybecystidiosa is distinguished by its growth on the leaf litter of coniferous forests, a flesh-pink pileus, a fibrous stipe with a white tomentose covering at the base, non-cellular pileipellis, larger basidiospores, and the presence of cheilocystidia. The reconstruction of phylogenetic trees using combined ITS, nLSU, and tef1-α sequences, employing maximum likelihood and Bayesian inference analyses, showed that C.betulicola formed a cluster with C.decurrens, while C.cystidiosa clustered with C.vinacea. However, these two clusters formed separate branches themselves, which also supported the results obtained from our morphological studies. A key to the Calocybe species reported from northeast China is provided to facilitate future studies of the genus.

General Synthesis of Conformationally Constrained Noncanonical Amino Acids with C(sp3)-Rich Benzene Bioisosteres.

Recent years have seen novel modalities emerge for the treatment of human diseases resulting in an increase in beyond rule of 5 (bRo5) chemical matter. As a result, synthetic innovations aiming to enable rapid access to complex bRo5 molecular entities have become increasingly valuable for medicinal chemists' toolkits. Herein, we report the general synthesis of a new class of noncanonical amino acids (ncAA) with a cyclopropyl backbone to achieve conformational constraint and bearing C(sp3)-rich benzene bioisosteres. We also demonstrate preliminary studies toward utilities of these ncAA as building blocks for medicinal chemistry research.

Soliton and dispersive wave generation with third-order dispersion and temporal boundary.

We investigate the pulse evolution and energy conservation condition at the temporal boundary under third-order dispersion. When the fundamental soliton crosses the temporal boundary and forms two reflected pulses and one transmitted pulse, the power of the transmitted pulse first increases and then decreases as the incident spectrum shifts toward the blue side. If the transmitted spectrum lies in the anomalous group-velocity dispersion region, second-order soliton is formed and dispersive wave is radiated. We present a modified phase-matching condition to predict the resonance frequencies. The predicted results are in good agreement with the results obtained by numerically solving the nonlinear Schrödinger equation.

Chronic cadmium exposure impairs flight behavior by dampening flight muscle carbon metabolism in bumblebees.

Cadmium pollution affects the global ecosystem because cadmium can be transferred up the food chain. The bumblebee, Bombus terrestris, is an important insect pollinator. Their foraging activity on flowers exposes them to harmful heavy metals, which damages their health and leads to massive population declines. However, the effects of chronic exposure to heavy metals on the flight performance of bumblebees have not yet been characterized. Here, we studied variation in the flight capacity of bumblebees induced by chronic cadmium exposure at field-realistic concentrations using behavioral, physiological, and molecular approaches. Chronic cadmium exposure caused a significant reduction in the duration, distance, and mean velocity of bumblebee flight. Transcriptome analysis showed that the impairment of carbon metabolism and mitochondrial dysfunction in the flight muscle were the primary causes. Physiological, biochemical, and metabolomic analyses validated disruptions in energy metabolism, and impairments in mitochondrial respiratory chain complexes activities. Histological analysis revealed muscle fiber damage and mitochondrial loss. Exogenous decanoic acid or citric acid partially restored sustained flight ability of bumblebees by mitigating muscle fiber damage and increasing energy generation. These findings provide insights into how long-term cadmium stress affects the flight ability of insects and will aid human muscle or exercise-related disease research.

Electron Microscope Study of the Pressure-Induced Phase Transformation and Microstructure Change of TiO2 Nanocrystals.

Microstructure transformation of materials under compression is crucial to understanding their high-pressure phase transformation. However, direct observation of the microstructure of compressive materials is a considerable challenge, which impedes the understanding of the relations among phase transformation, microstructure, and material properties. In this study, we used transmission Kikuchi diffraction and transmission electron microscopy to intuitively characterize pressure-induced phase transformation and microstructure of TiO2. We observed the changes of twin boundaries with increasing pressure and intermediate phase TiO2-I of anatase transformed into TiO2-II (α-PbO2 phase) for the first time. The following changes occur during this transformation: anatase (diameter of ∼100 nm) → anatase twins 60° along the [110] zone axis → intermediate TiO2-I twins 60° along the [010] zone axis → TiO2-II twins 90° along the [010] zone axis. These results directly reveal the crystallographic relation among these structures, enhancing our understanding of the phase transformation in TiO2 nanocrystals.

Alanine metabolism mediates energy allocation of the brown planthopper to adapt to resistant rice.

INTRODUCTION: During the adaptation to host plant resistance, herbivorous insects faced the challenge of overcoming plant defenses while ensuring their own development and reproductive success. To achieve this, a strategic allocation of energy resources for detoxification and ecological fitness maintenance became essential. OBJECTIVE: This study aimed to elucidate the intricate energy allocation mechanisms involved in herbivore adaptation that are currently poorly understood. METHODS: The rice Oryza sativa and its monophagous pest, the brown planthopper (BPH), Nilaparvata lugens were used as a model system. An integrated analysis of metabolomes and transcriptomes from different BPH populations were conducted to identify the biomarkers. RNA interference of key genes and exogenous injection of key metabolites were performed to validate the function of biomarkers. RESULTS: We found that alanine was one of the key biomarkers of BPH adaptation to resistant rice variety IR36. We also found that alanine flow determined the adaptation of BPH to IR36 rice. The alanine aminotransferase (ALT)-mediated alanine transfer to pyruvate was necessary and sufficient for the adaptation. This pathway may be conserved, at least to some extent, in BPH adaptation to multiple rice cultivars with different resistance genes. More importantly, ALT-mediated alanine metabolism is the foundation of downstream energy resource allocation for the adaptation. The adapted BPH population exhibited a significantly higher level of energy reserves in the fat body and ovary when fed with IR36 rice, compared to the unadapted population. This rendered the elevated detoxification in the adapted BPH and their ecological fitness recovery. CONCLUSION: Overall, our findings demonstrated the crucial role of ALT-mediated alanine metabolism in energy allocation during the adaptation to resistant rice in BPH. This will provide novel knowledge regarding the co-evolutionary mechanisms between herbivores and their host plants.

Investigation into the performance and stability of immobilized and coated polysaccharide columns in supercritical fluid chromatography.

In this study, the performance of the widely used "golden four" coated chiral stationary phases (Chiralpak AD-3, Chiralcel OD-3, Chiralpak AS-3, and Chiralcel OJ-3) was compared with their corresponding immobilized versions (Chiralpak IA-3, Chiralpak IB-3, Chiralpak IB N-3, Chiralpak IH-3, and Chiralpak IJ-3) under supercritical fluid chromatography (SFC) conditions with a set of 30 racemic compounds. Using the traditional modifiers, methanol and isopropanol, the immobilized columns (Chiralpak IB N-3 and Chiralpak IH-3) showed an improved general ability to successfully resolve the enantiomers of the target analytes relative to their coated versions (Chiralcel OD-3 and Chiralpak AS-3), while the coated columns (Chiralpak AD-3, Chiralcel OD-3, and Chiralcel OJ-3) performed better than their immobilized versions (Chiralpak IA-3, Chiralpak IB-3, and Chiralpak IJ-3). An investigation of the non-traditional modifiers, dichloromethane, ethyl acetate, and tetrahydrofuran with immobilized columns, revealed a generally decreased ability to successfully resolve the enantiomers of the target analytes, relative to the use of the traditional modifiers, methanol and isopropanol. The stability of the coated columns (Chiralpak AD-H and Chiralcel OD-H) was evaluated by injecting "forbidden" solvents, including dichloromethane, dimethyl sulfoxide, and tetrahydrofuran. After 200 injections of these solvents on coated columns, the retention factors and resolutions slightly decreased, and a significant increase in column backpressure was observed, indicating some degree of stationary phase degradation.

Analytical and preparative chiral supercritical fluid chromatography resolutions using crown ether-derived column.

In this study, crown ether-derived column Crownpak® CR-I (+) was evaluated under SFC conditions using 12 primary amines, and the chromatographic results were compared against eight immobilized polysaccharide-based columns. Crownpak® CR-I (+) achieved a significantly higher success rate. It was found that the addition of 5% water to the modifier dramatically improved the peak shape for chiral separation of primary amines on Crownpak® CR-I (+). The first reported preparative SFC separations on Crownpak® CR-I (+) are shown, offering a new approach for the preparative resolution of primary amines. The case studies demonstrate that Crownpak® CR-I (+) is a very useful column in the chiral separation of challenging compounds that contain a primary amine group in the pharmaceutical industry.