Shedding light on ONOO- detection: the emergence of a fast-response fluorescent probe for biological systems.

ONOO-, a bioactive molecule, plays a critical role in inflammation-related signaling pathways and pathological mechanisms. Numerous studies have established a direct correlation between elevated ONOO- levels and tumor progression. Therefore, investigating ONOO- levels in inflammation and tumors is of utmost importance. Fluorescence imaging presents a highly sensitive, non-invasive, easily operable, selective, and efficient method for ONOO- detection in situ. In this study, we designed and synthesized a rhodamine-based probe, NRho, which effectively identifies tumors, inflammatory cells, tissues, and organs by detecting ONOO- content. The synthesis process of NRho is simple, yielding a probe with favorable spectral characteristics and rapid response. Our cell imaging analysis has provided novel insights, revealing distinct ONOO- levels among different types of cancer cells, with hepatocellular carcinoma cells exhibiting higher ONOO- content than the others. This observation marks the proposal of such variations in ONOO- levels across cancer cell types. Furthermore, our study has showcased the practicality of our probe in live organ imaging, enabling the identification of tumors from living organs within a brief 5-minute incubation period. Additionally, our findings highlight the rapid detection capability of the probe NRho in various tissue samples, effectively identifying inflammation. This research holds important promise in advancing biomedical research and clinical diagnosis.

Mixed-Lineage Leukemia 1 Inhibition Enhances the Differentiation Potential of Bovine Embryonic Stem Cells by Increasing H3K4 Mono-Methylation at Active Promoters.

Mixed-lineage leukemia 1 (MLL1) introduces 1-, 2- and 3-methylation into histone H3K4 through the evolutionarily conserved set domain. In this study, bovine embryonic stem cells (bESCs, known as bESCs-F7) were established from in vitro-fertilized (IVF) embryos via Wnt signaling inhibition; however, their contribution to the endoderm in vivo is limited. To improve the quality of bESCs, MM-102, an inhibitor of MLL1, was applied to the culture. The results showed that MLL1 inhibition along with GSK3 and MAP2K inhibition (3i) at the embryonic stage did not affect bESCs' establishment and pluripotency. MLL1 inhibition improved the pluripotency and differentiation potential of bESCs via the up-regulation of stem cell signaling pathways such as PI3K-Akt and WNT. MLL1 inhibition decreased H3K4me1 modification at the promoters and altered the distribution of DNA methylation in bESCs. In summary, MLL1 inhibition gives bESCs better pluripotency, and its application may provide high-quality pluripotent stem cells for domestic animals.

Imaging and Detection of Hepatocellular Carcinoma with a Hepatocyte-Specific Fluorescent Probe.

Hepatocellular carcinoma is a highly invasive malignant tumor of the liver, which is the main cause of cancer-related death. The cancerization of hepatocytes may lead to the changes of cell microenvironment, active substances, and enzymes. Viscosity is one of the important parameters of cell microenvironment. Therefore, the study of the change in the viscosity of hepatocytes is very important for the detection and treatment of liver cancer. However, the hepatocyte-specific fluorescent probes which can detect viscosity have not been developed yet. Herein, the first hepatocyte-specific fluorescent probe (HT-V) for viscosity detection was designed and synthesized, which exhibited excellent optical properties for biological imaging studies. By using the unique probe HT-V, compared with the normal liver cells, a significant increase of viscosity in the liver cancer cells was observed in the cell imaging experiment. The organ imaging experiments showed that the probe HT-V could be successfully used to diagnose and image hepatocellular carcinoma in vivo. In addition, in situ imaging revealed that the new probe HT-V can specifically target and image hepatocellular carcinoma in mice. We expected that this powerful tool may provide guidance for the detection and imaging of hepatocellular carcinoma in the future.

Evaluation of Cell Viability with a Single Fluorescent Probe Based on Two Kinds of Fluorescence Signal Modes.

Accurate evaluation of cell viability is important for dosage tests of anticancer drugs, pathology, and numerous biological experiments. However, due to the serious insufficieny of fluorescent probes, which can distinguish various cell states, the study of cell viability is immensely limited. To resolve this issue, we design and synthesize a new probe ACD-E to monitor cell viability with two kinds of fluorescence signal modes, the first single fluorescent probe that can distinguish three different cell states and furnish accurate information in biological experiments. ACD-E can discriminate live and dead cells in a dual-color mode by evaluating cell mitochondrial esterase activity and can also discriminate live and early necrosis cells by determining mitochondrial viscosity in a "turn-on" mode in the near-infrared region. Significantly, the novel ACD-E can also distinguish cell viability in vivo. This work establishes a robust strategy for monitoring multiple cell states using a single fluorescent probe.

Adaptive intensity-modulated radiotherapy with simultaneous integrated boost for stage III non-small cell lung cancer: Is a routine adaptation beneficial?

PURPOSE: Tumor and anatomical changes during radiotherapy have been observed in stage III non-small cell lung cancer (NSCLC) from many previous studies. We hypothesized that a routinely scheduled adaptive radiotherapy would have clinical important dose benefits to lower the risk of toxicities, without increasing the tumor recurrences. METHODS: We retrospectively reviewed 92 consecutive patients with inoperable stage III NSCLC between November 2017 and March 2019. All eligible patients should received simultaneously integrated boost (SIB) using intensity-modulated radiation therapy (IMRT). A mid-treatment CT simulation and a new adapted plan were routinely given after the first 20 fractions. The organs at risk (OARs) were delineated per RTOG 1106 atlas. Dose-volume histograms were quantitatively compared between the initial and composite adaptive plans. Logistic regression was applied to analyze the dose-response relationship. Clinical endpoints included acute symptomatic radiation pneumonitis (RP2) and esophagitis (RE2), local and regional tumor control, and progression-free survival (PFS). RESULTS: Sixty-four eligible patients received adaptive SIB-IMRT were consecutively included. The GTVs reduced by a median of -38.2% after 42 to 44 Gy in 20 fractions of radiotherapy. By adapting to tumor and anatomical changes, dosimetric parameters of OARs decreased significantly. The mean lung dose decreased by an average of -74.8 cGy, and mean esophagus dose was lower by 183.1 cGy. We found grade 2 or higher acute RP in 11 patients (17.2%), and RE2 in 28 patients (43.8%). Commonly used lung and esophagus dose metrics were significantly associated with RP2 and RE2. The adaptation could reduce RP2 probability by 3%, and RE2 risk by 5%. Subgroups with higher OARs dose or larger tumor shrinkage may get more dose and toxicities benefits. The estimated median PFS was 12.5 months from the start of radiotherapy. CONCLUSIONS: We demonstrated that the routinely adaptive SIB-IMRT strategy could significantly reduce the dose to surrounding normal tissues, potentially lower the associated acute RP and RE, without increasing the risk of tumor recurrences.

A fluorescent probe for specific detection of ß-galactosidase in living cells and tissues based on ESIPT mechanism.

ß-galactosidase is of great significance to living organisms, which is an important marker of primary ovarian cancer and cellular senescence. To detect the activity of ß-galactosidase, a novel fluorescent probe ESIPT-GAL which based on excited state intramolecular proton transfer (ESIPT) mechanism for detecting ß-galactosidase is developed in this work with low background fluorescence and high sensitivity (ΦF = 0.0045-0.2409). The fluorescence intensity at 552 nm of this probe increased by ~ 55 times with ß-galactosidase addition (0-4 U/mL), and its detection limit is very low (3.9 × 10-5 U/mL). In addition, the spectral data (pseudo-first-order rate: 1.303 min-1) and enzyme kinetic parameter (Vmax = 69.5 µΜ•S-1) both show that the probe can achieve rapid response to ß-galactosidase. Moreover, the probe has good water solubility, which ensures that it has good biocompatibility and can be easily applied to detect ß-galactosidase in living cells and tissues. Importantly, the probe ESIPT-GAL can monitor ß-galactosidase in deep mouse tissue sections (90 µm).

MLL1 combined with GSK3 and MAP2K inhibition improves the development of in vitro-fertilized embryos.

The MM-102 compound prevents the interaction between mixed lineage leukemia 1 (MLL1) and WD Trp-Asp repeat domain 5 (WDR5) and results in the inhibition of MLL1 H3K4 histone methyltransferase (HMT) activity. The inhibition of the FGFR signaling pathway and activation of the WNT pathway by small molecule inhibitors (known as 2i) improves blastocyst development. However, studies on the effects of MLL1 combined with GSK3 and MAP2K inhibition (3i) on the development of embryos have not been reported. Our results show that 3i improves bovine and mouse IVF development only when added at the appropriate time point and affects ICM-related gene (OCT4, SOX2 and NANOG) expression in a concentration-dependent manner. 3i increases the expression of blastocyst-related genes such as PRDM14, KLF4 and KLF17 and decreases the expression of the de novo DNA methyltransferase genes DNMT3L and DNMT1 in bovines, but increases Prdm14, Stella, Klf2 and Klf4 expression and significantly decreases Dnmt3l, Dnmt3b, and Dnmt1 expression in mice. The analysis of transcription data showed that the expression of DNMTs increases slightly later than that of PRDM14 during embryo development, which indicates that PRDM14 is the upstream regulator. 3i upregulates PRDM14 and then downregulates DNMTs to affect IVF embryo development. When 3i-treated mouse embryos were transplanted, the morphology and body weight of the offspring were not significantly different from those of the control group. These offspring were as fertile as normal mice. 3i improves the development of bovine and mouse IVF embryos but does not affect the quality of the embryos. The application of 3i provides a new method for improving IVF embryo production in domestic animals.

[Study on prevalence and correlation factors of bronchial asthma in Zaozhuang area, Shandong province].

OBJECTIVE: To study the prevalence of asthma and its correlated factors in Zaozhuang area in 2003, to provide a basic consideration for prevention/treatment and control policy. METHODS: 6 points were selected by stratified-clusterd-random sampling with a total of 16,725 persons expected, but only 10,610 subjects investigated. RESULTS: In this survey, 128 asthma cases were identified with a overall prevalence of 1.21%. The prevalence for children was 2.02%, and for adult was 0.90% with the former significantly higher then the latter (chi(2) = 21.39, P < 0.01). Rates for male and female were 1.08%, 1.32% with a ratio of 1:1.22. For 77.97% of children with asthma. The initiative age of asthma was before 7 years old among children while among 36.23% of the adults, it was before 15 years of age. Correlation analysis showed that upper respiratory tract infection (OR = 17.81, 95% CI: 12.25-25.89), cold air exposure (OR = 3.43, 95% CI: 2.41-4.90), stimulation through cooking and by harmful gases (OR = 2.56, 95% CI: 1.80-3.63), allergic materials (OR = 2.74, 95% CI: 1.80-4.17) were main inducing factors. 65.63% of the asthma cases having had history of allergic disease while 25.78% having had family history with the OR of allergic history and family history as 21.69 vs. 73.96. CONCLUSION: The epidemic status of bronchial asthma was serious, with an assumption that asthma cases might have reached the number of 43 thousand in Zaozhuang area.