10-Methoxycamptothecin (MCPT), a natural camptothecin derivative, shows notable anticancer activity. Using UPLC-QTOF-MS, this study characterized MCPT metabolite profiles in rats, detecting 14 metabolites in urine, 5 in bile, 7 in feces, and 2 in plasma. The findings indicated that phase I metabolites of MCPT were primarily formed via demethylation, demethoxylation, and hydroxylation, while phase II metabolites mainly included glucuronide and sulfate of MCPT and its phase I metabolites. MCPT, hydroxycamptothecin, and the subsequent glucuronide conjugates were the predominant forms of MCPT in rats. This study provided a comprehensive overview of the metabolite profile of MCPT in rats.

Background:Sustained release of bupivacaine can be achieved by encapsulating bupivacaine within multivesicular liposomes, providing localised analgesia for up to 72 h. This study aimed to evaluate whether a multimodal analgesic approach integrating epidural analgesia with liposomal bupivacaine enhanced transversus abdominis plane block could extend the interval to initial opioid use and thus reduce the total post-operative opioid requirements in women undergoing caesarean section. Methods:Women scheduled for elective caesarean delivery under combined spinal-epidural anaesthesia were randomly assigned to a liposomal bupivacaine or placebo group. All participants were given 0.75% ropivacaine 15 mg intrathecally at L3-L4 interspace. Before surgical closure, 0.6 mg of epidural hydromorphone was administered. Bilateral ultrasound-guided lateral transversus abdominis plane blocks were performed after surgery. The liposomal bupivacaine group received 133 mg liposomal bupivacaine in 20 mL fluid per side, whereas the placebo control group received 20 mL saline per side. Postoperative analgesia included scheduled oral acetaminophen and self-administered boluses of oxycodone as needed. The primary outcomes were 24- and 48-h oxycodone consumption. Results:A total of 128 women were enrolled. The median [interquartile range (IQR)] postoperative cumulative oxycodone consumption was significantly lower in the liposomal bupivacaine group than in the placebo group at 24 h [2 (0-5) mg vs. 4 (1-8) mg, p = 0.009] and 48 h [8 (0-13) mg vs. 10 (4-18) mg, p = 0.022], and the median (IQR) interval to first patient-controlled analgesia use was significantly longer in the liposomal bupivacaine group than in the placebo group [22 (12-48) vs. 8 (4-18) h; p < 0.001]. Conclusion:Epidural hydromorphone combined with liposomal bupivacaine enhances transversus abdominis plane block, prolongs analgesia duration and reduces opioid requirements in the first 48 h after caesarean-section. Clinical trial registration:https://www.chictr.org.cn/showproj.html?proj=237618, Identifier [ChiCTR2400087477].

Leaf vein, an essential part of leaf architecture, plays significant roles in shaping the proper leaf size. To date, the molecular mechanisms governing leaf development including leaf venation patterning remains poorly understood in birch. Here, we performed the genome-wide identification of homeodomain-like (HD-like) superfamily genes using phylogenetic analysis and revealed the functional role of a potential HD-like gene in leaf growth and development using transgenic technology and transcriptomic sequencing. A total of 267 HD-like genes were identified based on Arabidopsis HD-containing transcription factors, which were members of KNOTTED1-like homeobox (KNOX) family, BELL1-like homeobox (BLH) family, Zinc finger-HD (ZHD) family, HD-leucine zipper (HD-Zip) family, Golden2, ARR-B, Psr1 (GARP) family, WUSCHEL-related homeobox (WOX) family, and Myeloblastosis (MYB) and MYB-like family. Further, 41 HD-like genes showing co-expression with marker genes related to leaf vascular tissues exhibited differential expression during primary vein development. Among them, a potential HD-like gene (BpPHD4) of GARP family served as a negative factor in governing leaf size and venation patterning. Compared to non-transgenic plants, BpPHD4 repression transgenic plants showed increased leaf length, leaf width, leaf area, leaf thickness, spongy tissue thickness, stomata number, epidermal cell size, primary vein length, the distance between the secondary veins, and primary vein diameter, which was opposite to those of BpPHD4 overexpression transgenic plants. Meanwhile, reduced expression levels of BpPHD4 could remarkably promote phloem tissue development. Transcriptome analysis of BpPHD4 overexpression transgenic plants showed two candidate genes (Bpev01.c0518.g0018 and Bpev01.c2797.g0002) probably regulated by BpPHD4. To conclude, our findings contribute to a better understanding of HD-like superfamily genes and unravel the role of a potential HD-like gene in genetically controlling leaf size and venation patterning in birch, which provides clues to genetic improvement of woody plants with diverse geometric and topological properties of leaf vascular network.

Betulinic acid (BA) is a plant-derived pentacyclic triterpenoid with various biological properties. Studies have identified that the deprotonated molecular ion of BA could not generate any product ions by an LC-ESI/MS/MS. In this study, BA reacted with p-toluenesulfonyl isocyanate (PTSI) to produce an ionizable and fragmentary derivative for LC-ESI/MS/MS detection in a negative mode. Thus, we developed and validated an UPLC-ESI/MS/MS method to quantitate BA in rat plasma. The calibration curve was linear over the concentration range of 5–1280 ng/mL for BA, with a lower limit of detection of 1 ng/ml. This validated method was successfully applied to a pharmacokinetic study of BA in rat plasma after intravenous administration of BA.

Low-grade glioma (LGG) can affect any part of the central nervous system. This study aimed to investigate the mechanism of Aidi injection (AIDI) in treating LGG using network pharmacology, The Cancer Genome Atlas, and molecular docking technology. Open databases were used to obtain AIDI targets and components as well as LGG genes. To identify essential elements and targets, a protein-protein interaction network and an active ingredient-common target network were built. Kyoto encyclopedia of genes and genomes and gene ontology enrichment analysis were used to screen the major signaling pathways. The clinical importance of key genes was confirmed using both survival and differential expression studies. The top 6 components in terms of degree value on the active ingredient-common target network were considered as the core components of AIDI for treating LGG. CASP3, TGFβ1, TP53, and EGFR were termed hub target genes, which are not only aberrantly expressed in LGG tissue, but their excessive expression was also found to lower patient survival rates. The biological processes linked to the antitumor activity of AIDI primarily involve responses to hormones and reactive oxygen species. Kyoto encyclopedia of genes and genomes enrichment analysis identified PI3K-AKT and MAPK as the most important signaling pathways. Molecular docking experiments demonstrated strong binding between the hub genes and the essential components of AIDI. AIDI may target CASP3, TGFβ1, TP53, and EGFR and modulates PI3K-AKT and MAPK signaling pathways to exert its anticancer action against LGG. Reactive oxygen species may play an important role in this process.

Traditional anti-corrosion and anti-fouling coatings struggle against the harsh marine environment. Our study tackled this by introducing a novel dual-layer hydrogel (A-H DL) coating system. This system combined a Cu2O–SiO2–acrylic resin primer for anchoring and controlled copper ion release with a dissipative double-network double-anchored hydrogel (DNDAH) boasting superior mechanical strength and anti-biofouling performance. An acrylamide monomer was copolymerized and cross-linked with a coupling agent to form the first irreversible network and first anchoring, providing the DNDAH coating with mechanical strength and structural stability. Alginate gel microspheres (AGMs) grafted with the same coupling agent formed the second reversible network and second anchoring, while coordinating with Cu2+ released from the primer to form a system buffering Cu2+ release, enabling long-term antibacterial protection and self-healing capabilities. FTIR, SEM, TEM, and elemental analyses confirmed the composition, morphology, and copper distribution within the A-H DL coating. A marine simulation experiment demonstrated exceptional stability and anti-fouling efficacy. This unique combination of features makes A-H DL a promising solution for diverse marine applications, from ship hulls to aquaculture equipment.

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Polygonatum cyrtonema polysaccharides have a variety of pharmacological effects. The commonly used extraction methods include traditional hot water extraction, alkaline extraction, enzymatic hydrolysis method, ultrasonic-assisted extraction, etc., but there are problems such as low yield, high temperature, high cost, strict extraction conditions, and insufficient environmental protection. In this study, crude polysaccharide extraction from the Polygonatum cyrtonema Hua was performed using the freeze–thaw method. Response surface methodology (RSM), based on a three-level, three-variable Box–Behnken design (BBD), was employed to obtain the best possible combination of water-to-raw material ratio (A: 30–50), freezing time (B: 2–10 h), and thawing temperature (C: 40–60 °C) for maximum polysaccharide extraction. Using the multiple regression analysis and analysis of variance (ANOVA), the experimental data were fitted to a second-order polynomial equation and were used to generate the mathematical model of optimization experiments. The optimum extraction conditions were as follows: a water-to-raw material ratio of 36.95:1, a freezing time of 4.8 h, and a thawing temperature of 55.99 °C. Under the optimal extraction conditions, the extraction rate of Polygonatum cyrtonema Hua polysaccharide (PCP) was 65.76 ± 0.32%, which is well in close agreement with the value predicted by the model, 65.92%. In addition, PCP has significant antioxidant activity. This result shows that the freeze–thaw method can improve the extraction efficiency, maintain the structural integrity of polysaccharides, simplify the extraction process, promote the dispersion of polysaccharides, and is suitable for large-scale industrial production.

Camptothecin (CPT) and its derivatives are potent candidates for cancer treatment. However, the clinical ap-plications are largely restricted by non-selectivity and severe toxicities. The peptide transporter 1 (PEPT1), which is highly expressed in human intestines, has been found to be overexpressed in several cancer cells. This dis-covery suggests that PEPT1 has the potential to serve as a therapeutic target for both improving bioavailability and cancer-targeting treatment. Therefore, a prodrug approach for CPT targeting at PEPT1 highly expressed cancer cells was adopted in the present study. Eighteen CPT prodrugs, its peptidic conjugates, were synthesized and the structures were confirmed by NMR and HRMS. The protein expression profiles of PEPT1 in different cell lines were performed using immunofluorescence assay and western blotting analysis. The cytotoxicity of CPT prodrugs and their uptake via competition with Gly-Sar, a typical substrate of PEPT1, were evaluated in both PEPT1-overexpressed and under expressed cells. The results demonstrated that most CPT prodrugs significantly impaired Gly-Sar uptake, suggesting a higher affinity of CPT-peptidic conjugates for PEPT1 and PEPT1 over-expression cells. In addition, these prodrugs demonstrated a higher capability for inhibiting cell growth in PEPT1 highly-expressed cancer cells compared to PEPT1 under expressed cells. These results indicated that this peptidic prodrug strategy might offer great potential for improved tumor selectivity and chemotherapeutic efficacy of CPT.

Betulinic acid (BA) and betulin (BE) are naturally pentacyclic triterpenes with documented biological activities, especially antitumor and anti-inflammatory activity. However, their bioavailability in vivo is not satisfactory in terms of medical applications. Thus, to improve the solubility and bioavailability so as to improve the efficacy, 28-O-succinyl betulin (SBE), a succinyl derivative of BE, was synthesized and its solubility, in vitro and in vivo anti-tumor activities, the apoptosis pathway as well as the pharmacokinetic properties were investigated. The results showed that SBE exhibited significantly higher solubility in most of the tested solvents, and showed a maximum solubility of 7.19 ± 0.66 g/L in n-butanol. In vitro and in vivo anti-tumor activity assays indicated both BA and SBE exhibited good anti-tumor activities, and SBE demonstrated better potential compared to BA. An increase in the ratio of Bad/Bcl-xL and activation of caspase 9 was found in SBE treated Hela cells, suggesting that the intrinsic mitochondrial pathway is involved in SBE induced apoptosis. Compared with BA, SBE showed much-improved absorption and bioavailability in pharmacokinetic studies.

Diabetic nephropathy is a major complication of diabetes mellitus and is related to dysfunction of renal cells. Hederagenin is a triterpenoid saponin from some Chinese herbs with anti-inflammatory and anti-diabetic activities. However, its role in diabetic nephropathy progression is still obscure. This study aimed to explore the effects of hederagenin on renal cell dysfunction in vitro. Human renal mesangial cells (HRMCs) and human renal proximal tubular epithelial cells (HRPTEpiCs) were cultured under high glucose (HG) conditions to mimic diabetic nephropathy-like injury. Cell proliferation was evaluated by CCK-8. mRNA and protein levels were determined by qRT-PCR and western blotting, respectively. The secretion levels of fibrosis-related biomarkers were analyzed by ELISA. Results showed that hederagenin reduced HG-induced proliferation increase in HRMCs and HRPTEpiCs. Hederagenin attenuated HG-induced increase in mRNA and protein expression of NLRP3, ASC, and IL-1β. Hederagenin also suppressed HG-induced increase in mRNA and secretion levels of FN, Col. IV, PAI-1, and TGF-β1. NLRP3 inhibitor MCC950 attenuated HG-induced fibrosis of renal cells, and its activator nigericin reversed the suppressive effect of hederagenin on HG-induced fibrosis. Bioinformatics analysis predicted cathepsin B (CTSB) as a target of hederagenin to modulate NOD-like receptor (NLR) pathway. Hederagenin decreased CTSB level, and CTSB overexpression reversed the suppressive effect of hederagenin on HG-induced NLRP3 inflammasome activation and fibrosis in HRMCs and HRPTEpiCs. In conclusion, hederagenin attenuates HG-induced fibrosis of renal cells by inhibiting NLRP3 inflammasome activation via reducing CTSB expression, indicating a therapeutic potential of hederagenin in diabetic nephropathy.

BackgroundPrussian blue (PB) nanoparticles (NPs) have been intensively investigated for medical applications, but an in-depth toxicological investigation of PB NPs has not been implemented. In the present study, a comprehensive investigation of the fate and risks of PB NPs after intravenous administration was carried out by using a mouse model and an integrated methodology of pharmacokinetics, toxicology, proteomics, and metabolomics.ResultsGeneral toxicological studies demonstrated that intravenous administration of PB NPs at 5 or 10 mg/kg could not induce obvious toxicity in mice, while mice treated with a relatively high dose of PB NPs at 20 mg/kg exhibited loss of appetite and weight decrease in the first two days postinjection. Pharmacokinetic studies revealed that intravenously administered PB NPs (20 mg/kg) underwent fast clearance from blood, highly accumulated in the liver and lungs of mice, and finally cleared from tissues. By further integrated proteomics and metabolomics analysis, we found that protein expression and metabolite levels changed significantly in the liver and lungs of mice due to the high accumulation of PB NPs, leading to slight inflammatory responses and intracellular oxidative stress.ConclusionsCollectively, our integrated experimental data imply that the high accumulation of PB NPs may cause potential risks to the liver and lungs of mice, which will provide detailed references and guidance for further clinical application of PB NPs in the future.

The core-shell nano drug loaded particle DOX@AuNC@HA???DAH???was prepared by a simple one potmethod using gold nanocage???AuNC???as the core???sulfhydryl modified hyaluronic acid???LC-HA???as the shell and doxo???rubicin hydrochloride???DOX???as the drug model.The gold nanocage provides a container for drug loading and endowsthe carrier with photothermal properties.The modified hyaluronic acid encapsulates the gold nanocage and providespH/enzyme response and targeted mediating functions.The structure of DAH was characterized???and the drugloading???controlled release performances???cell uptake and cytotoxicity were studied.The results show that the DAHnanoparticles with the core-shell structures have a high drug loading capacity???a good cycle stability and excellentphotothermal conversion efficiency under the irradiation of laser source.In phosphate buffer solution with a pH of7.4???DAH has high stability???the DOX leakage ratio in20h is less than20%???and DAH exhibits good stimulusresponse and can release the loaded drug quickly in acidic???hyaluronidase and photothermal action environment.In addition???DAH is more easily uptaken by tumor cells in cell experiments???which demonstrates certain targeting???showing better stimulus responsiveness.When chemotherapy and photothermal therapy works together???the activity oftumor cells is greatly weakened???showing the advantages and potential of combined therapy.

White birch ( Betula platyphylla ) is precious material for pulpwood and widely distributed in 14 provinces of China. Previous study indicated that inhibited expression of a gene encoding an auxin amide synthase, BpGH3.5 , in transgenic plants reduced the level of IAA–amino acid conjugation, resulting in more free IAA, thereby better growth of birch. Utilizing transgenic- BpGH3.5 lines to increase wood production in a wide range of environments is the goal for breeders. In three field trials here, we measured tree height, diameter at breast height, and volume of 16 BpGH3.5 -transgenic 7-year-old white birch lines (including 12 antisense strand lines and 4 overexpression lines) and a wild-type white birch line from three sites that varied greatly in their environmental conditions. To select elite BpGH3.5- transgenic lines for each target environment, we used an additive main effects and multiplicative interaction model to analyze genotype by environment interaction, growth adaptability and stability. The selection criteria for elite transgenic lines were set as the average volume plus 0.75 times the standard deviation for the tested lines at each test site. Results showed that the effect of line and site for height was highly significant ( P < 0.01), and the effect of line × site was significant ( P < 0.05); selected as the elite lines were FG12, FG13 and FG27 at the Maoershan Experimental Forest Farm, FG13 and FG32 at the Shidaohe Forest Farm, and FG3 and FG31 at the Ecological Experiment Forest Farm. These seven high-yield, stable lines can now be tested in production trials or adjacent trial areas with similar environmental conditions, while the high-yield, unstable lines should be tested in production trials in areas deemed suitable for their growth. These results provide guidance on which released transgenic elite lines will grow best in a wide range of conditions.

本文以贵州省雷公山国家森林公园为研究对象,采用文献查阅、实地调研、问卷调查相结合的方法,分析其森林康养产业的发展现状,总结发展中存在的问题,参考其总体要求,针对问题提出科学可行的对策建议,以期促进雷公山国家森林公园森林康养产业走出一条极具当地特色的发展之路,为贵州省乃至全国其他森林康养基地发展森林康养提供借鉴.

Multiple independent sequence variants of the hTERT locus have been associated with telomere length and cancer risks in genomewide association studies. Here, we identified an intronic variable number tandem repeat, VNTR2-1, as an enhancer-like element, which activated hTERT transcription in a cell in a chromatin-dependent manner. VNTR2-1, consisting of 42-bp repeats with an array of enhancer boxes, cooperated with the proximal promoter in the regulation of hTERT transcription by basic helix-loop-helix transcription factors and maintained hTERT expression during embryonic stem-cell differentiation. Genomic deletion of VNTR2-1 in MelJuSo melanoma cells markedly reduced hTERT transcription, leading to telomere shortening, cellular senescence, and impairment of xenograft tumor growth. Interestingly, VNTR2-1 lengths varied widely in human populations; hTERT alleles with shorter VNTR2-1 were underrepresented in African American centenarians, indicating its role in human aging. Therefore, this polymorphic element is likely a missing link in the telomerase regulatory network and a molecular basis for genetic diversities of telomere homeostasis and age-related disease susceptibilities.

Root rot of Populus davidiana × P. alba var. pyramidalis Louche (Pdpap) is caused by Fusarium oxysporum. We used RNA sequencing to study the molecular mechanisms and response pattern of Pdpap infected by F. oxysporum CFCC86068. We cloned the PdpapWRKY28 transcription factor gene and transformed the recombinant vector pBI121-PdpapWRKY28 into Pdpap. The resistance function of PdpapWRKY28 was verified using physiological and biochemical methods. By means of RNA sequencing, we detected 1,403 differentially expressed genes (DEGs) that are common in the different treatments by F. oxysporum. Furthermore, we found that overexpression of the PdpapWRKY28 gene may significantly improve the resistance of Pdpap plants to F. oxysporum. Our research reveals a key role for PdpapWRKY28 in the resistance response of Pdpap to F. oxysporum. Additionally, our results provide a theoretical basis for in-depth research on resistance breeding to combat root rot.

Late-embryogenesis-abundant proteins (LEA) typically accumulate to high levels during seed dehydration and have an important role in the plant response to abiotic stresses. Transgenic hybrid poplars of 9-year-old TaLEA lines showed adaptation to drought and saline–alkali conditions. The exogenous gene was expressed normally, and the T-DNA insertion sites were identified on different chromosomes. Improving the survival rate of afforested species is essential for forest production in saline–alkali areas. Transgenic hybrid poplars could provide potential genetic materials for adaptation to semi-arid areas. We investigated the growth stability and adaptability of 9-year-old TaLEA transgenic lines of Populus simonii × nigra, as well as the position of T-DNA insertion and the stability of exogenous genes. Transgenic poplar field trials were carried out using a multi-site joint analysis. Molecular characteristics and the T-DNA insertion sites were identified by genome resequencing. The XL-9, XL-7, and XL-13 preferred lines consistently showed the highest and most stable growth across three testing sites. The exogenous gene was integrated into the genome chromosome and was expressed normally in the XL-1, XL-7, and XL-9 lines after 9 years. XL-9, XL-7, and XL-13 are the preferred lines for subsequent production tests. Transgenic hybrid poplar lines can be selected for adaptability 2 years after field experiments. The results of this study will provide further guidance for testing of transgenic trees for future forest production.