Pd-Catalyzed C(sp2)-C(sp3) Suzuki Coupling and Synthesis of Lumacaftor Using Designed Monophosphine Ligands.

Novel monophosphine ligands L1 and L2 with a C-P ring were designed and synthesized for the efficient Pd-catalyzed C(sp2)-C(sp3) Suzuki coupling. With 0.5 mol % Pd2dba3 and 2 mol % L1, aryl halides coupled with alkylboronic acids to give the products in up to 99% yield. The aryl thianthrene salt was further applied for late-stage methylation in 97% yield. The active pharmaceutical ingredient lumacaftor was synthesized by aryl-alkyl and aryl-aryl Suzuki coupling reactions using L1 and L2.

Phosphorus modifies the association between body mass index and uric acid: Results from NHANES 2007-2018.

INTRODUCTION: Studies in recent years have shown that high uric acid causes harm to the human body, which has become a serious public health problem. Elevated serum uric acid has been shown to be associated with obesity, but the relationship between BMI and uric acid (UA) remains controversial. Although the association between BMI and UA has been well studied, the effect of phosphorus levels in vivo on this association remains unclear. This study aimed to determine the relationship between BMI and serum uric acid and the effect of phosphorus on the relationship between the two. RESEARCH DESIGN AND METHODS: The present study analyzed data from the National Health and Nutrition Examination Survey (NHANES) continuous 2007-2018 cycle. We included 10786 participants aged 20 years and over. Multivariable linear regression was performed to assess the association between BMI and serum uric acid. phosphorus was stratified into low phosphorus (<3.3 mg/dl), middle phosphorus (3.3-3.9 mg/dl) and high phosphorus (>3.9 mg/dl). Correction of the effect of phosphorus was assessed by testing the interaction between BMI and UA in multivariate linear regression. RESULTS: In this cross-sectional study, we found that BMI was positively associated with UA in the female population but not significantly in the male population or in the total population. In multiple regression analysis, UA was 0.51 higher in the highest female BMI group than in the lowest group (p = 0.0001). The relationship between BMI and UA differed significantly by gender under the influence of phosphorus, with men and women in Model II having a greater elevation of UA in men than in women within most groups. (BMI >30, phosphorus >3.9 mg/dl, ß:0.83 95% CI: 0.43, 1.23 vs ß: 0.79 95% CI: 0.30, 1.29). In addition, phosphorus significantly altered the positive association between BMI and UA in most models. CONCLUSION: Our results indicate significant associations between BMI and uric acid in women, with higher BMI values likely to be associated with a higher risk of hyperuricemia, suggesting that uric acid levels in obese people should be closely monitored in clinical practice. Phosphorus and BMI have an interactive effect in elevating UA and should be noted as indicators of phosphorus in clinical practice.

Bruton's tyrosine kinase inhibition suppresses pathological retinal angiogenesis.

BACKGROUND AND PURPOSE: Pathological retinal angiogenesis is a typical manifestation of vision-threatening ocular diseases. Many patients exhibit poor response or resistance to anti-vascular endothelial growth factor (VEGF) agents. Bruton's tyrosine kinase (BTK) controls the proliferation and function of immune cells. Therefore, we examined the anti-inflammatory and anti-angiogenic effects of BTK inhibition on retinal angiogenesis. EXPERIMENTAL APPROACH: Retinal neovascularisation and vascular leakage in oxygen-induced retinopathy in C57/BL6J mice were assessed by whole-mount retinal immunofluorescence. PLX5622 was used to deplete microglia and Rag1-knockout mice were used to test the contribution of lymphocytes to the effects of BTK inhibition. The cytokines, activation markers, inflammatory and immune-regulatory activities of retinal microglia/macrophages were detected using qRT-PCR and immunofluorescence. NLRP3 was detected by western blotting, and the effects of BTK inhibition on the co-culture of microglia and human retinal microvascular endothelial cells (HRMECs) were examined. KEY RESULTS: BTK inhibition suppressed pathological angiogenesis and vascular leakage, and significantly reduced retinal inflammation, which involved microglia/macrophages but not lymphocytes. BTK inhibition increased anti-inflammatory factors and reduced pro-inflammatory cytokines that resulted from NLRP3 inflammasome activation. BTK inhibition suppressed the inflammatory activity of microglia/macrophages, and acted synergistically with anti-VEGF without retinal toxicity. Moreover, the supernatant of microglia incubated with BTK-inhibitor reduced the proliferation, tube formation and sprouting of HRMECs. CONCLUSION AND IMPLICATIONS: BTK inhibition suppressed retinal neovascularisation and vascular leakage by modulating the inflammatory activity of microglia and macrophages. Our study suggests BTK inhibition as a novel and promising approach for alleviating pathological retinal angiogenesis.

Pharmacologic evaluation of delayed long-acting cabotegravir administration among cisgender women in HPTN 084.

HPTN 084 demonstrated the superiority of long-acting injectable cabotegravir (CAB-LA) compared with daily oral tenofovir disoproxil fumarate-emtricitabine (F/TDF) for HIV prevention in women. CAB-LA (600 mg) or placebo injections were administered 4 weeks after an initial dose (loading dose) and every 2 months (Q2M) thereafter; this is the approved regimen. Participants experienced both loading dose and Q2M delays during the trial. CAB concentrations were evaluated before a delay, at the visit associated with the delay, and the visit after a delayed injection was administered. During the blinded phase of the trial, 194 participants randomized to CAB-LA experienced at least one injection delay. Plasma CAB concentrations were maintained above the 4× protein adjusted 90% inhibitory concentration (4× PA-IC90; protocol-specific threshold) for all loading dose and 98% of Q2M delays when injections were administered up to 6 weeks late. The feasibility of shifting to an every 3-month (Q3M) regimen in females was interrogated via simulation studies using a population pharmacokinetic model. Q3M injections in both CAB-naïve (with a loading dose) and previously CAB-exposed females were predicted to yield higher steady-state exposures than in males on the approved Q2M regimen. Although there is observed forgiveness following an isolated delayed CAB-LA injection and simulations suggest acceptable CAB-LA exposures in women with a 600 mg CAB-LA Q3M regimen, empirical efficacy of this regimen has not been established, and transitioning to this dosing schema is not recommended. Future pharmacokinetic bridging studies are aimed at evaluating higher dose CAB-LA formulations administered less frequently. CLINICAL TRIALS: This study is registered with ClinicalTrials.gov as NCT03164564.

Crystal structure of exfoliative toxin D from Staphylococcus aureus.

Staphylococcus aureus exfoliative toxins (ETs) are serine proteases responsible for staphylococcal scalded skin syndrome. Four ETs, ETA, ETB, ETD, and ETE, have been identified, all of which cleave desmoglein-1. This study presents the crystal structure of ETD at 1.75 Å resolution. The protein exhibits a structure composed of two ß-barrels and two α-helices as described in previous studies of ETs. A predicted model of ETD in complex with Ile380-Glu381-Gly382-Pro383 (IEGP), a segment of human desmoglein-1 (hDsg1), was constructed. Glu381 of hDsg1 was predicted to interact with as many as six amino acid residues in ETD, whereas two amino acid residues in ETD primarily constituted subsite S1', and a space near subsite S1' was noted. It is likely that polypeptide chains located near the IEGP segment in the predicted structure of hDsg1 bind to this space. The structure of loop D, which was predicted to participate in subsite S2', in ETD was markedly different from those in other ETs.

Deep Learning for Distinguishing Mucinous Breast Carcinoma From Fibroadenoma on Ultrasound.

PURPOSE: Mucinous breast carcinoma (MBC) tends to be misdiagnosed as fibroadenomas (FA) due to its benign imaging characteristics. We aimed to develop a deep learning (DL) model to differentiate MBC and FA based on ultrasound (US) images. The model could contribute to the diagnosis of MBC for radiologists. METHODS: In this retrospective study, 884 eligible patients (700 FA patients and 184 MBC patients) with 2257 US images were enrolled. The images were randomly divided into a training set (n = 1805 images) and a test set (n = 452 images) in a ratio of 8:2. First, we used the training set to establish DL model, DL+ age-cutoff model and DL+ age-tree model. Then, we compared the diagnostic performance of three models to get the optimal model. Finally, we evaluated the diagnostic performance of radiologists (4 junior and 4 senior radiologists) with and without the assistance of the optimal model in the test set. RESULTS: The DL+ age-tree model yielded higher areas under the receiver operating characteristic curve (AUC) than DL model and DL+ age-cutoff model (0.945 vs. 0.835, P < .001; 0.945 vs. 0.931, P < .001, respectively). With the assistance of DL+ age-tree model, both junior and senior radiologists' AUC had significant improvement (0.746-0.818, P = .010, 0.827-0.860, P = .005, respectively). CONCLUSIONS: The DL+ age-tree model based on US images and age showed excellent performance in the differentiation of MBC and FA. Moreover, it can effectively improve the performance of radiologists with different degrees of experience that may contribute to reducing the misdiagnosis of MBC.

Global Analysis of Natural Products Biosynthetic Diversity Encoded in Fungal Genomes.

Fungal secondary metabolites (SMs) represent an invaluable source of therapeutic drugs. Genomics-based approaches to SM discovery have revealed a vast and largely untapped biosynthetic potential within fungal genomes. Here, we used the publicly available fungal genome sequences from the NCBI public database, as well as tools such as antiSMASH, BIG-SLiCE, etc., to analyze a total of 11,598 fungal genomes, identifying 293,926 biosynthetic gene clusters (BGCs), which were subsequently categorized into 26,825 gene cluster families (GCFs). It was discovered that only a tiny fraction, less than 1%, of these GCFs could be mapped to known natural products (NPs). Some GCFs that only contain a single BGC internally are crucial for the biodiversity of fungal biosynthesis. Evident patterns emerged from our analysis, revealing popular taxa as prominent sources of both actual and potential biosynthetic diversity. Our study also suggests that the genus rank distribution of GCF is generally consistent with NP diversity. It is noteworthy that genera Xylaria, Hypoxylon, Colletotrichum, Diaporthe, Nemania, and Calonectria appear to possess a higher potential for SM synthesis. In addition, 7213 BGCs match possible known compound structures, and homologous gene clusters of well-known drugs can be located in different genera, facilitating the development of derivatives that share structural similarity to these drugs and may potentially possess similar biological activity. Our study demonstrated the various types of fungi with mining potential, assisting researchers in prioritizing their research efforts and avoiding duplicate mining of known resources to further explore fungal NP producers.

Tryptophan-rich diet and its effects on Htr7+ Tregs in alleviating neuroinflammation and cognitive impairment induced by lipopolysaccharide.

BACKGROUND: Neuroinflammation is a vital pathogenic mechanism for neurodegenerative diseases such as Alzheimer's, schizophrenia, and age-related cognitive decline. Regulatory T cells (Tregs) exhibit potent anti-inflammatory properties and can modulate neurodegenerative diseases arising from central nervous system inflammatory responses. However, the role of Tregs in neuroinflammation-related cognitive dysfunction remains unclear. It is highly plausible that Htr7+ Tregs expressing unique genes associated with the nervous system, including the Htr7 gene encoding the serotonin receptor 5-HT7, play a pivotal role. METHODS: Mice were given a tryptophan-rich diet (with a tryptophan content of 0.6%) or a normal diet (with a tryptophan content of 0.16%). The neuroinflammation-mediated cognitive dysfunction model was established by intracerebroventricular injection of lipopolysaccharide (LPS) in 8-week-old C57BL/6J mice. The activation and infiltration of Tregs were measured using flow cytometry. Primary Tregs were cocultured separately with primary CD8+ T cells and primary microglia for in vitro validation of the impact of 5-HT and 5-HT7 receptor on Tregs. Prior to their transfer into recombination activating gene 1 (Rag1-/-) mice, Tregs were ex vivo transfected with lentivirus to knock down the expression of Htr7. RESULTS: In this study, the tryptophan-rich diet was found to reverse LPS-induced cognitive impairment and reduce the levels of 5-HT in peripheral blood. The tryptophan-rich diet led to increased levels of 5-HT in peripheral blood, which in turn promoted the proliferation and activation of Htr7+ Tregs. Additionally, the tryptophan-rich diet was also shown to attenuate LPS-mediated neuroinflammation by activating Htr7+ Tregs. Furthermore, 5-HT and 5-HT7 receptor were found to enhance the immunosuppressive effect of Tregs on CD8+ T cells and microglia. In Rag1-/- mice, Htr7+ Tregs were shown to alleviate LPS-induced neuroinflammation and cognitive impairment. CONCLUSIONS: Our research revealed the ability of Htr7+ Tregs to mitigate neuroinflammation and prevent neuronal damage by suppressing the infiltration of CD8+ T cells into the brain and excessive activation of microglia, thereby ameliorating LPS-induced cognitive impairment. These insights may offer novel therapeutic targets involving Tregs for neuroinflammation and cognitive impairment.

Novel multidomain peptide self-assembly biomaterials based on bola structure and terminal anchoring: Nanotechnology meets antimicrobial therapy.

To ameliorate the diminished antimicrobial efficiency and physiological stability associated with monomeric antimicrobial peptides (AMPs) molecules, future research will focus on the artificial design of self-assembling peptides to replace monomeric entities, aiming to combat the antibiotic resistance crisis caused by microbial infections. In this study, the "bola" structure was used as the foundational architecture driving molecular self-assembly, with hydrophobic amino acids at the termini to anchor and finely adjust the sequence, thereby organizing a range of novel multidomain peptides (MDPs) templates into an ABA block motif. The results indicate that FW2 (GMSI = 53.94) exhibits the highest selectivity index among all MDPs and can form spherical micelles in an aqueous medium without the addition of any exogenous additives. FW2 exhibited high stability in vitro in the presence of physiological salt ions, serum, and various pH conditions. It exhibited excellent biocompatibility and efficacy both in vivo and in vitro. Furthermore, FW2 strongly interacts with the lipid membrane and employs various synergistic mechanisms, such as reactive oxygen species (ROS) accumulation, collectively driving cellular apoptosis. This study demonstrates a straightforward strategy for designing self-assembling peptides and promotes the advancement of peptide-based biomaterials integration progress with nanotechnology.

Studying the effects of Saposhnikoviae Radix on the pharmacokinetic profiles of 10 bioactive compounds originating from Astragali Radix in rat plasma by UHPLC-QTRAP-MS/MS.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragali Radix-Saposhnikoviae Radix (AR-SR) is a well-known and effective herb pair. Although the compatibility of these two herbs has been widely applied in many traditional Chinese medicine formulas, its potential mechanism still needs to be investigated. AIM OF STUDY: To evaluate the pharmacokinetic profiles of 10 bioactive compounds derived from AR when administrated alone and in combination with SR to rats, aiming to further reveal the impact of SR on AR. MATERIALS AND METHODS: Two groups of male Sprague-Dawley rats received oral administration of AR and AR-SR freeze-dried powder solutions, respectively. UHPLC-QTRAP-MS/MS technology was utilized to perform the pharmacokinetic studies of 10 compounds derived from AR in rat plasma samples. RESULTS: A reliable UHPLC-QTRAP-MS/MS method was established to simultaneously determine the rat plasma concentrations of eight isoflavonoids, referring to calycosin (CAL), calycosin-7-O-ß-D-glucoside (CAL-G), formononetin (FOR), formononetin-7-O-ß-D-glucoside (FOR-G), astrapterocarpan (APC), astrapterocarpan-3-O-ß-D-glucoside (APC-G), astraisoflavan-7-O-ß-D-glucoside (AIF-G) and formononetin-7-O-ß-D-glucuronide (FOR-GN), and two saponins, including astragaloside IV (AS IV) and cycloastragenol (CAG), originating from AR. Following the oral administration of AR, seven isoflavonoids were quickly absorbed but exhibited low plasma concentrations under 17.88 ng/mL except FOR-GN. The latter maintained higher plasma concentration level more than 15 ng/mL for at least 10 h. Besides, for the first time, AS IV was observed with an obvious double-peak phenomenon after administering AR extract, whereas the concentration of CAG was lower than LLOQ before 6 h. When AR and SR were administrated together, the double-peak phenomena of CAL, FOR, APC, AIF-G and FOR-GN were enhanced and there was a significant increase in their values of area under the concentration-time curve (AUC) and mean residence time (MRT) (P < 0.05) while the pharmacokinetic profiles of CAL-G, FOR-G, APC-G, AS IV and CAG stayed almost unchanged (P > 0.05). Moreover, the elimination half-time (t1/2) values of CAL, FOR and APC were significantly elevated, and the clearance rate/bioavailability (CLz/F) for CAL and FOR was reduced (P < 0.05). CONCLUSIONS: SR has the potential to modulate the ADME process of five out of the eight isoflavonoids (CAL, FOR, APC, AIF-G and FOR-GN, except CAL-G, FOR-G and APC-G) originating from AR. This interaction is especially likely to affect the hepatic and intestinal drug disposition of these isoflavonoids, thereby extending the duration of their pharmacological effects, which may subsequently impact the therapeutic efficacy of AR.

Sub-kHz-linewidth broadband-swept fiber laser based on Rayleigh scattering-enabled Brillouin random lasing oscillation with dynamic tunning Brillouin gain spectrum.

A sub-kHz-linewidth broadband-swept fiber laser using Rayleigh scattering-based Brillouin random lasing oscillation is proposed and experimentally demonstrated. Benefiting from Brillouin-involved acoustic damping and arbitrary-wavelength distributed Rayleigh feedback, leveraging instantaneously tuning Brillouin gain spectrum induced by a frequency-sweeping pump, a highly coherent random lasing emission with cavity mode elimination as well as frequency noise suppression is achieved in a sweeping manner. Results show that the proposed sweeping Stokes laser with a two-order-magnitude compressed linewidth of 840 Hz and 20 dB frequency noise suppression can unprecedentedly operate over the maximum wavelength range of 16 nm. Dynamic characteristics of the sweeping laser frequency are experimentally investigated, indicating a minimum residual nonlinearity of 0.0001 within the frequency-sweeping range of 126.63 GHz. It is believed that the proposed swept fiber laser may have attractive potential in diverse applications, including sensing and imaging.

Recent progress on triterpenoid derivatives and their anticancer potential.

Cancer poses a significant global public health challenge, with commonly used adjuvant or neoadjuvant chemotherapy often leading to adverse side effects and drug resistance. Therefore, advancing cancer treatment necessitates the ongoing development of novel anticancer agents with diverse structures and mechanisms of action. Natural products remain crucial in the process of drug discovery, serving as a primary source for pharmaceutical leads and therapeutic advancements. Triterpenoids are particularly compelling due to their complex structures and wide array of biological activities. Recent research has demonstrated that naturally occurring triterpenes and their derivatives have the potential to serve as promising candidates for new drug development. This review aims to comprehensively explore the anticancer properties of triterpenoids and their synthetic analogs, with a focus on recent advancements. Various aspects, such as synthesis, phytochemistry, and molecular simulation for structure-activity relationship analyses, are summarized. It is anticipated that triterpenoid derivatives will emerge as notable anticancer agents following further investigation into their mechanisms of action and in vivo studies.

Controlled release characteristics of alkyl gallates and gallic acid from ß-cyclodextrin inclusion complexes of alkyl gallates.

The freeze-drying approach was used to create inclusion complexes utilizing alkyl gallates and ß-cyclodextrin, namely dodecyl gallate, octyl gallate, butyl gallate, and ethyl gallate, which are exemplary examples of phenolic esters. The everted-rat-gut-sac model demonstrated that the inclusion complexes released alkyl gallates, which were subsequently hydrolyzed to generate free gallic acid, as evidenced by HPLC-UV analysis. Both gallic acid and short-chain alkyl gallates were capable of permeating the small intestinal membrane. The transport rate of gallic acid (or alkyl gallates) exhibited an initial rise followed by a drop when the carbon-chain lengths varied. The inclusion complex groups exhibited a superior sustained-release effect compared to the comparable alkyl gallates groups, thus possibly leading to higher bioavailability and stronger bioactivity. Moreover, altering the length of the carbon chain will allow for the effortless achievement of regulated release of phenolic compounds and short-chain phenolic esters from such ß-cyclodextrin inclusion complexes.

Combining PCT with CRP is better than separate testing for patients with bacteriuria in the intensive care unit: a retrospective study.

BACKGROUND: Previous studies on PCT for urinary tract infections (UTI) have focused primarily on minors. This study investigated the predictive value of the neutrophil-to-lymphocyte ratio (NLR), C-reactive protein (CRP) level and procalcitonin (PCT) level in adult patients with bacteriuria in IUC. METHODS: This case‒control study included 85 patients with bacteriuria (PB) in the ICU from March 2021 to Jan 2024 based on positive urine culture results and a control group (n = 136) from Jan 2024 to March 2024. Patient data were collected using a hospital information management system. ROC curves of the NLR, CRP and PCT were use to predict the PB. RESULTS: The AUCs of the NLR, CRP and PCT for the prediction of PB in ICU were 0.711 (95% CI 0.644-0.772), 0.855 (95% CI 0.800-0.900), and 0.884 (95% CI 0.832-0.924), respectively; the optimal thresholds were 8.02, 18.52 mg/L, and 0.215 ng/mL, respectively; the sensitivities were 69.0 (95% CI 56.9-79.5), 90.1 (95% CI 80.7-95.9), and 83.1 (95% CI 72.3-91.0), respectively; and the specificities were 67.6 (95% CI 59.1-75.4), 68.4 (95% CI 59.9-76.1), and 80.9 (95% CI 73.3-87.1), respectively. The negative predictive value (NPV) of CRP is greater than that of PCT. In bacteriuria caused by Candida infections, CRP and PCT have higher sensitivity and NPV. CONCLUSIONS: Combined CRP and PCT testing is more helpful for diagnosing bacteriuria. CRP and PCT have higher sensitivity and NPV in diagnosing bacteriuria caused by Candida infection.

Electrical Stimulation Promotes Endocytosis of Magnetic Nanoparticles by Cancer Cells.

Nanomaterials are increasingly used in biomedical imaging and cancer therapy, and how to improve the endocytosis of nanomaterials by cells is a key issue. The application of alternating current (AC) electrical stimulation to osteosarcoma cells (MG-63) here can increase the cellular endocytosis of Fe3O4 nanoparticles (diameter: 50 nm) by 52.46% via macropinocytosis. This can be ascribed to the decrease in F-actin content and the increase in intracellular Ca2+ concentration. Transmission electron microscope, immunofluorescence staining, western blot, flow cytometry, and inductively coupled plasma emission spectrometer analyses support this interpretation. The application of electrical stimulation decreases the cell viability in magnetic hyperthermia by 47.6% and increases the signal intensity of magnetic resonance imaging by 29%. Similar enhanced endocytosis is observed for breast cancer cells (MCF-7), glioblastoma cells (U-87 MG), melanoma cells (A-375), and bladder cancer cells (TCCSUP), and also for Fe3O4 nanoparticles with the diameters of 20 and 100 nm, and Zn0.54Co0.46Cr0.65Fe1.35O4 nanoparticles with the diameter of 70 nm. It seems the electrical stimulation has the potential to improve the diagnostic and therapeutic effects of magnetic nanoparticles by promoting endocytosis.

Hydrogeochemical features, genesis, and quality appraisal of confined groundwater in a typical large sedimentary plain.

The confined groundwater of arid sedimentary plains has been disturbed by long-term anthropogenic extraction, and its hydrochemical quality is required for sustainable development. The present research investigates the hydrochemical characteristics, formation, potential health threats, and quality suitability of the confined groundwater in the central North China Plain. Results show that the confined groundwater has a slightly alkaline nature in the study area, predominantly dominated by fresh-soft Cl-Na and HCO3-Na types. Water chemistry is governed by water-rock interactions, including dissolution of evaporites and cation exchange. Approximately 97% of the sampled confined groundwaters exceed the prescribed standard for F-. It is mainly due to geological factors such as mineral dissolution, cation exchange, and competitive adsorption of HCO3 - and may also be released from compacted soils because of groundwater extraction. Enriched F- in the confined groundwater can pose an intermediate and higher non-carcinogenic risk to more than 90% of the population. It poses the greatest health threat to the population in the north-eastern part of the study area, especially to infants and children. For sustainable development, the long-term use of confined groundwater for irrigation in the area should be avoided, and attention should also be paid to the potential soil salinization and infiltration risks. In the study area, 97% of the confined groundwaters are found to be excellent or good quality for domestic purposes based on Entropy-weighted Water Quality Index. However, the non-carcinogenic health risk caused by high contents of F- cannot be ignored. Therefore, it is recommended that differential water supplies should be implemented according to the spatial heterogeneity of confined groundwater quality to ensure the scientific and rational use of groundwater resources. PRACTITIONER POINTS: The hydrochemistry quality of confined groundwater in an arid sedimentary plain disturbed by long-term anthropogenic extraction was investigated. The suitability of confined groundwater for multiple purposes such as irrigation and drinking were evaluated. The hydrochemical characteristics and formation mechanism of confined groundwater under the influence of multiple factors were revealed.

Effects of Zishen Yutai pills combined with metformin on women with polycystic ovary syndrome undergoing in vitro fertilization.

In this study, we analyzed the clinical efficacy of Zishen Yutai pills (ZSYTP) combined with metformin hydrochloride on infertile women diagnosed with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization and embryo transfer (IVF-ET). Patients were assigned into 3 groups: the ZSYTP group (n = 50), the metformin group (n = 50), and the combination group (ZSYTP combined with metformin hydrochloride, n = 50), based on their respective and the indicated treatments before undergoing IVF-ET. Then, their glucose metabolism indices, sex hormone indices, traditional Chinese medicine (TCM) syndrome scores, and outcomes of IVF-ET were compared. Baseline characteristics were not significantly different between the 2 groups. After treatment, various parameters such as body mass index (BMI), fasting plasma glucose (FPG), fasting insulin (FIN), homeostatic model assessment of insulin resistance (HOMA-IR), luteinizing hormone (LH), estradiol (E2), follicle-stimulating hormone (FSH), testosterone (T) levels, and TCM syndrome scores were found to be reduced compared to pretreatment levels in both groups. Moreover, the improvement observed in the treatment group exceeded that of the control group. Specifically, the observation group displayed significantly lower gonadotropin (Gn) dosage and duration, as well as a reduced abortion rate compared to the control group. Furthermore, the observation group had higher numbers of obtained eggs, high-quality embryos, eggs obtained through IVF-ET, average transferred embryos, clinical pregnancy rate, and embryo implantation rate compared to the control group. Pretreatment with ZSYTP combined with metformin before IVF-ET in PCOS patients improves the outcome of IVF-ET.

Metarhizium spp. encode an ochratoxin cluster and a high efficiency ochratoxin-degrading amidohydrolase revealed by genomic analysis.

INTRODUCTION: Ochratoxins (OTs) are worldwide regulated mycotoxins contaminating a variety of food-environment and agro-environment. Several Aspergillus and Pencillium species synthesize OTs from a six-gene biosynthetic gene cluster (BGC) to produce the highly toxic final product OTA. Although many studies on OTA-degrading enzymes were performed, high efficiency enzymes with strong stability are extremely needed, and the OTA degrading mechanism is poorly understood. OBJECTIVES: The study aimed to explore the OT-degradation enzyme and investigate its degradation mechanisms in Metarhizium, which contain an OT biosynthetic gene cluster. METHODS: Phylogenomic relationship combined with RNA expression analysis were used to explore the distribution of OT BGC in fungi. Bioactivity-guided isolation and protein mass spectrometry were conducted to trace the degrading enzymes in Metarhizium spp., and the enzymes were heterologously expressed in E. coli and verified by in vitro assays. Structure prediction and point mutation were performed to reveal the catalytic mechanism of MbAmh1. RESULTS: Beyond Aspergillus and Pencillium species, three species of the distant phylogenetic taxon Metarhizium contain an expressed OT-like BGC but lack an otaD gene. Unexpectedly, no OT BGC products were found in some Metarhizium species. Instead, Metarhizium metabolized both OTA and OTB to their non-toxic degradation products. This activity of M. brunneum was attributed to an intracellular hydrolase MbAmh1, which was tracked by bioactivity-guided proteomic analysis combined with in vitro reaction. Recombinant MbAmh1 (5 µg/mL) completely degraded 1 µg/mL OTA within 3 min, demonstrating a strong degrading ability towards OTA. Additionally, MbAmh1 showed considerable temperature adaptability ranging from 30 to 70 °C and acidic pH stability ranging from 4.0 to 7.0. Identification of active sites supported the crucial role of metal iron for this enzymatic reaction. CONCLUSION: These findings reveal different patterns of OT synthesis in fungi and provide a potential OTA degrading enzyme for industrial applications.

Multi-omics profiling of retinal pigment epithelium reveals enhancer-driven activation of RANK-NFATc1 signaling in traumatic proliferative vitreoretinopathy.

During the progression of proliferative vitreoretinopathy (PVR) following ocular trauma, previously quiescent retinal pigment epithelial (RPE) cells transition into a state of rapid proliferation, migration, and secretion. The elusive molecular mechanisms behind these changes have hindered the development of effective pharmacological treatments, presenting a pressing clinical challenge. In this study, by monitoring the dynamic changes in chromatin accessibility and various histone modifications, we chart the comprehensive epigenetic landscape of RPE cells in male mice subjected to traumatic PVR. Coupled with transcriptomic analysis, we reveal a robust correlation between enhancer activation and the upregulation of the PVR-associated gene programs. Furthermore, by constructing transcription factor regulatory networks, we identify the aberrant activation of enhancer-driven RANK-NFATc1 pathway as PVR advanced. Importantly, we demonstrate that intraocular interventions, including nanomedicines inhibiting enhancer activity, gene therapies targeting NFATc1 and antibody therapeutics against RANK pathway, effectively mitigate PVR progression. Together, our findings elucidate the epigenetic basis underlying the activation of PVR-associated genes during RPE cell fate transitions and offer promising therapeutic avenues targeting epigenetic modulation and the RANK-NFATc1 axis for PVR management.

The diagnostic value of 68Ga-NOTA-MAL-Cys-MZHER2:342 PET/CT imaging for HER2-positive lung adenocarcinoma.

Background: The human epidermal growth factor receptor 2 gene (HER2) has been identified as a potential therapeutic target in lung adenocarcinoma (LUAD). Non-invasive positron emission tomography (PET) imaging provides a reliable strategy for in vivo determination of HER2 expression through whole-body detection of abnormalities. The PET tracer 68Ga-NOTA-MAL-Cys-MZHER2:342 has shown promising results for HER2-positive breast and gastric cancers. This study aims to evaluate the performance of 68Ga-NOTA-MAL-Cys-MZHER2:342 in vitro and in vivo models and in clinical patients with HER2-positive LUAD. Methods: NOTA-MAL-Cys-MZHER2:342 was synthesized and labeled with 68Ga. Cell uptake, cell binding ability, and stability studies of 68Ga-NOTA-MAL-Cys-MZHER2:342 were assessed both in the Calu-3 lung cancer (LC) cell line and normal mice. In vivo assessment in tumor-bearing mice was conducted using microPET imaging and biodistribution experiments. Additionally, preliminary PET/CT imaging analysis was performed on HER2-positive LC patients. Results: 68Ga-NOTA-MAL-Cys-MZHER2:342 was prepared with a radiochemical purity (RCP) exceeding 95%. The tracer demonstrated high cell uptake in HER2-overexpressing Calu-3 cells, with an IC50 of 158.9, an adequate 1.73 nM. Good stability was exhibited both in vitro and in vivo. MicroPET imaging of Calu-3-bearing mice revealed high tumor uptake and notable tumor-to-background ratios. Positive outcomes were also observed in two HER2-positive LUAD patients. Conclusion: 68Ga-NOTA-MAL-Cys-MZHER2:342 demonstrated satisfactory stability, sensitivity, and specificity. These findings suggest that 68Ga-NOTA-MAL-Cys-MZHER2:342 PET/CT imaging provides a novel tool for non-invasive visual assessment of HER2 expression in LUAD patients.