The Chinese keratoconus (CKC) cohort study.

The Chinese keratoconus (CKC) cohort study is a population-based longitudinal prospective cohort study in the Chinese population involving a clinical database and biobanks. This ongoing study focuses on the prevention of KC progression and is the first to involve the effect of gene‒environment interactions on KC progression. The CKC cohort is hospital-based and dynamic and was established in Zhengzhou, China; KC patients (n = 1114) from a large geographical area were enrolled from January 2019 to June 2023, with a mean age of 22.23 years (6‒57 years). Demographic details, socioeconomic characteristics, lifestyle, disease history, surgical history, family history, and visual and social function data are being collected using questionnaires. General physical examination, eye examination, biological specimen collection, and first-degree relative data were collected and analyzed in the present study. The primary focus of the present study was placed on gene, environment and the effect of gene‒environment interactions on KC progression. The follow-up of the CKC cohort study is expected to include data collection at 3 months, 6 months, and 1 year after the initial examination and then at the annual follow-up examinations. The first follow-up of the CKC cohort study was recorded. A total of 918 patients completed the follow-up by June 1, 2023, with a response rate of 82.40%. Aside from the younger age of patients who were followed up, no significant differences were found between patients who were followed up and patients who were not.

Novel Artificial Scaffold for Bone Marrow Regeneration: Honeycomb Tricalcium Phosphate.

Bone marrow is complex structure containing heterogenetic cells, making it difficult to regenerate using artificial scaffolds. In a previous study, we succeeded in developing honeycomb tricalcium phosphate (TCP), which is a cylindrical scaffold with a honeycomb arrangement of straight pores, and we demonstrated that TCP with 300 and 500 µm pore diameters (300TCP and 500TCP) induced bone marrow structure within the pores. In this study, we examined the optimal scaffold structure for bone marrow with homeostatic bone metabolism using honeycomb TCP. 300TCP and 500TCP were transplanted into rat muscle, and bone marrow formation was histologically assessed. Immunohistochemistry for CD45, CD34, Runt-related transcription factor 2 (Runx2), c-kit single staining, Runx2/N-cadherin, and c-kit/Tie-2 double staining was performed. The area of bone marrow structure, which includes CD45(+) round-shaped hematopoietic cells and CD34(+) sinusoidal vessels, was larger in 300TCP than in 500TCP. Additionally, Runx2(+) osteoblasts and c-kit(+) hematopoietic stem cells were observed on the surface of bone tissue formed within TCP. Among Runx2(+) osteoblasts, spindle-shaped N-cadherin(+) cells existed in association with c-kit(+)Tie-2(+) hematopoietic stem cells on the bone tissue formed within TCP, which formed a hematopoietic stem cell niche similar to as in vivo. Therefore, honeycomb TCP with 300 µm pore diameters may be an artificial scaffold with an optimal geometric structure as a scaffold for bone marrow formation.

[Bile acid derivatives-focused chemical profiling in snake bile].

As a precious traditional Chinese medicine(TCM), snake bile has been widely used in numerous Chinese medicine prescriptions. Bile acid(BA) derivatives have been demonstrated as the primary chemical family in snake bile. In-depth chemical characterization of BAs is of great importance towards the establishment of quality standards and clarification of the effective material basis for snake bile. This study firstly employed ~1H-NMR to preliminarily analyze the chemical profiles of snake bile, an automated fraction collector was subsequently implemented to obtain the fractions-of-interest. The fraction was then concentrated and re-analyzed by LC-MS. Based on ~1H-NMR, BAs were found to be the main components of snake bile, and six BAs including CDCA, CA, TCDCA, TCA, TDCA and GCA were tentatively identified from the representative spectrum with the assistance of literature and reference compounds. Whereas the content of TCA in snake bile was too great, resulting in a great obstacle for the detection of trace components, the automated fraction collector was subsequently implemented to obtain the fractions-of-interest for LC-MS analysis. According to matching MS/MS information and retention time with reference compounds as well as database retrieval, a total of 57 BAs were detected and annotated. Because of the combination of ~1H-NMR and LC-MS platforms, the findings are beneficial for the in-depth characterization of BAs in snake bile, which provides references for the establishment of quality control and evaluation methods of snake bile.

ß-Lactams as promising anticancer agents: Molecular hybrids, structure activity relationships and potential targets.

ß-Lactam, commonly referred as azetidin-2-one, is a multifunctional building block for synthesizing ß-amino ketones, γ-amino alcohols, and other compounds. Besides its well known antibiotic activity, this ring system exhibits a wide range of activities, attracting the attention of researchers. However, the structurally diverse ß-lactam analogues as anticancer agents and their different molecular targets are poorly discussed. The purpose of this review is 3-fold: (1) to explore the molecular hybridization approach to design ß-lactams hybrids as anticancer agents; (2) the structure activity relationship of the most active anticancer ß-lactams and (3) to summarize their antitumor mechanisms.

[Direct chemome profiling of Boschniakia rossica using online pressurized extraction-ultrahigh-performance liquid chromatography-ion trap-time-of-flight mass spectrometry].

An online pressurized extraction-ultrahigh-performance liquid chromatography-ion trap-time-of-flight mass spectrometry (online PLE-UHPLC-IT-TOF-MS) platform was configured for the rapid and direct profiling of the chemical constituents of Boschniakia rossica (B. rossica). Notably, only a sub-microgram amount (0.5 mg) of B. rossica was placed in an empty guard column core, which was then filled up with normal-phase silica gel as an extraction vessel. The guard column core was placed inside a guard column holder in the column oven (70℃) and connected to the UHPLC-IT-TOF-MS system via metal tubing. The extraction phase was maintained for 3 min with 0.1% (v/v) formic acid as the extraction solvent. The extraction and elution phases for the entire measurement were segmented by a 6-port/2-channel electronic valve. In the elution phase, 0.1% (v/v) aqueous formic acid-acetonitrile containing 0.1% (v/v) formic acid was the mobile phase, and the components of interest that accumulated at the front of the column were eluted into the IT-TOF-MS system for detection. A total of 48 compounds were observed, and according to the literature, database, and mass fragmentation pathways, 45 compounds, including 10 phenylethanol glycosides, 14 iridoid glycosides, and 21 phenylalanine glycosides, were identified from B. rossica. This study provides reliable information regarding the chemical composition and quality evaluation of B. rossica. Moreover, it offers a promising analytical pipeline for the chemical composition characterization of traditional Chinese medicines.

Optimal collision energy is an eligible molecular descriptor to boost structural annotation: An application for chlorogenic acid derivatives-focused chemical profiling.

Tandem mass spectrometry (MS/MS), in particular high-resolution MS/MS, is able to provide element compositions and substructures for the detected signals. However, it is still challenging to configure the whole structures via linking those substructures. Efforts were devoted here to propose and validate optimal collision energy (OCE) to be an auxiliary structural clue to mass-to-charge ratios (m/z), and online energy-resolved MS was developed to yield OCEs. Chlorogenic acid derivatives (CADs) were utilized as the proof-of-concept because diverse isomers usually initiated by the different linkage manners between the quinic acid/shikimic acid and cinnamoyl substituents(s), i.e. caffeoyl group, coumaroyl group, etc. Liquid chromatography-hybrid ion trap-time of flight MS (LC-IT-TOF-MS) was implemented to capture CADs in two well-known herbal medicines namely Lonicerae japonicae Flos and Inulae Flos. Afterwards, hybrid triple quadrupole-linear ion trap MS (Qtrap-MS) was deployed to acquire OCEs for the primary fragmentation pathways of all detected CADs through online energy-resolved MS. On the other side, structural calculations were conducted to figure out the relationships between OCEs and bond properties. Isomeric differences occurred for OCEs, and LC elution program as well as ionization parameters could not affect OCEs. Twenty-four and thirty-one CADs were hunted and putatively identified by LC-IT-TOF-MS in Lonicerae japonicae Flos and Inulae Flos, respectively, and the structural annotation was advanced by applying the OCE-bond property relationships. To verify the structures, CADs-of-interest were purified from Lonicerae japonicae Flos using an automated fraction collector and definitely identified with NMR spectroscopy. Exact consistence occurred for the structural identification of mono-caffeoylquinic acid isomers between LC-MS/MS and NMR analyses. Consequently, OCE is an inherent physicochemical parameter of a given compound and is an eligible structural descriptor to offset the ability of LC-MS/MS towards the chemical profiling of complex matrices.

Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors.

Activated platelets release adenosine trisphosphate (ATP) and bioluminescence analysis of ATP release is usually used to monitor activation of platelets induced by various stimulants. However, bioluminescence analysis of ATP possesses poor linearity, the signal is quickly attenuated, and the accuracy of ATP release from platelets is hard to determine accurately enough to be used in a high throughput screening of platelet inhibitors. The present study was designed to optimize bioluminescence analysis of ATP released by platelets and expand its application in high throughput screening of platelet inhibitors. The results showed that accuracy of ATP analysis was significantly improved by adding coenzyme A (CoA) and signal attenuation of ATP analysis was greatly postponed by adding bovine serum albumin (BSA) both in Hank's balanced salt solution (HBSS) and Tyrode's buffer. Furthermore, ATP release of activated platelets and inhibitory effects of Ly294002 and Staurosporine on platelet activation were accurately determined by our optimized bioluminescence analysis of ATP. Thus, we have successfully constructed an optimized bioluminescence analysis of ATP which can be used in high throughput screening of platelet inhibitors.