Rapid epidemiological screening for tuberculosis (TB) usually uses tuberculin pure protein derivative (PPD) skin test, which has limitations such as low specificity and high side effects. ESAT-6 and CFP-10 are secreted proteins of Mycobacterium tuberculosis, but the related genes are missing from Bacillus Calmette-Guerin (BCG). In this study, the fusion protein ESAT6-CFP10 (EC) was expressed and purified, and prepared into chitosan nanoparticles (EC-NPs), which were loaded into the microneedle patch and carried out the preliminary test of tuberculosis skin test. The drug loading capacity of MNP-EC-NPs (microneedle patch, MNP) can reach 0.03 μg per needle and 1.92 μg per patch. The shelf life of MNP-EC-NPs can reach 6 months at room temperature, and it can effectively penetrate the epidermis. Volunteer skin test results showed that MNP-EC-NPs can effectively distinguish between BCG vaccinators, and can effectively show positive reaction in the skin of tuberculosis patients without significant side effects. The experiment was approved by the Ethics Committee of Wuhan Pulmonary Hospital [2022 (2)]. In this study, a TB skin test method was established, using ESAT6-CFP10 fusion protein instead of PPD, and using soluble microneedle dosage form, which improved the specificity of TB skin test diagnosis and provided a new technical scheme for TB epidemic screening.

Photothermal therapy is a new type of tumor therapy that uses near-infrared laser to specifically activate the photothermal agent accumulated in the lesion site, so as to achieve thermal ablation of cancer cells. However, the long metabolic cycle and difficult clearance of photothermal agent materials in vivo are also one of the major obstacles hindering their clinical transformation. In this work, we used hemoglobin as a novel stabilizer, hexachloroplatinic acid as a novel oxidant, and pyrrole as a monomer to prepare hemoglobin-stabilized platinum-based polypyrrole nanoparticles (Hb@PtP) by a one-step oxidative polymerization method and investigated in detail to study their physicochemical properties, such as morphology and structure. After polyethylene glycol modification, the particle size of obtained Hb@PtPP was 99.08 ± 8.3 nm and the zeta potential was -18.7 ± 1.2 mV. Transmission electron microscopy showed that the Hb@PtPP nanoparticles exhibited irregular spherical shape and uniform dispersion in the system. Under the irradiation of 808 nm laser with different power densities, Hb@PtPP showed power density-dependent temperature-raising behavior, and CCK-8 and dead-live staining experiments confirmed that they could effectively exert the photothermal effect to kill tumor cells. Small living animal imaging results demonstrated that Hb@PtPP had good tumor targeting and retention ability, and achieved tumor growth inhibition and ablation in vivo under laser excitation. All animal experiments involved in the study were performed in accordance with the program approved by the Animal Care and Use Committee of the Chinese Academy of Medical Sciences, Beijing Union Medical College, Institute of Radiation Medicine [IRM/2-IACUC-2312-005].

A high performance liquid chromatography (HPLC) method utilizing correction factors was established for the quantitative detection of related substances in flumazenil. Separation was achieved using an Agilent Pursuit XRs C18 column (250 mm × 4.6 mm, 5 μm) with an isocratic elution of dilute phosphoric acid, methanol, and tetrahydrofuran as the mobile phases. Correction factors calculated from a standard curve method were applied to determine the impurity content. The quantification of impurities in flumazenil was conducted using both external standard and correction factor methods, followed by validation and comparison of the two. For the identification of degradation products, a forced degradation approach was employed to prepare a flumazenil degradation solution, and the resulting impurities were confirmed by LC-MS analysis. The separation of flumazenil and its impurities was found to be efficient. The limits of quantification for impurities A, B, D, and E were established at 0.169 9, 0.314 7, 0.143 9, and 0.270 8 ng, respectively, with the limits of detection at 0.055 8, 0.096 9, 0.048 8, and 0.089 0 ng. These impurities demonstrated a strong linear relationship across the concentration ranges of 0.034 9-7.847 0, 0.038 7-8.710 7, 0.034 6-7.794 1, and 0.032 4-7.292 8 µg·mL^-1, respectively (n = 7). The method achieved average recoveries between 98.25% and 99.42%, with an RSD of less than 2.0% (n = 9), indicating high accuracy. The external standard and correction factor methods were used to determine the related substances in flumazenil, and the results of the two methods were consistent. The established HPLC method is characterized by its high accuracy, sensitivity, and repeatability, and is suitable for determining related substances in flumazenil.

Nephrotic syndrome (NS) has a variety of classifications, pathogenesis and pathological types. Clinical diagnosis primarily relies on serum biochemistry, while the specific classification necessitates renal puncture for biopsy, which is hindered by poor patient compliance. Therefore, it is of great significance for clinical diagnosis to find a non-invasive and rapid method to reflect the classification and progression of nephrotic syndrome. In this study, LC-MS metabolomics combined with receiver operating characteristic (ROC) and multiple linear regression analysis was used to screen and identify potential biomarkers capable of reflecting the typing and progression of nephrotic syndrome. According to the statistical parameters VIP > 1, P < 0.05 and AUC > 0.5 obtained from the orthogonal partial least squares discriminant analysis (OPLS-DA) model, five potential classification markers were screened to distinguish membranous nephropathy (MN) from IgA nephropathy (IgAN), including indoleacetic acid, isoleucine proline, DL-indole-3-lactic acid, D-phenylalanine and L-tryptophan. Furthermore, using estimated glomerular filtration rate (eGFR) as the dependent variable, a multiple linear regression analysis was conducted to identify the potential progression markers capable of reflecting the progression of MN to uremia. These metabolites included alanylleucine, 9-capryloylcarnitine, gluconic acid, caprylyl glycine and sebacic acid. Potential markers of progression of IgA nephropathy to uremia comprised alanylleucine, 9-capryloylcarnitine, caprylyl glycine, and sebacic acid. This study provides a theoretical basis for the discovery of potential classification and progression biomarkers of kidney disease, and also offers a methodological reference for future research in this area. The protocol was approved by the Ethics Committee of Shanxi Provincial People's Hospital [(2020) Provincial Medical Ke Lun Shen Zi No. 30].

This study aimed to investigate the role and mechanism of sappanone A (SA) in regulating renal ischemia-reperfusion injury (IRI) in rats. The animal experiment has been approved by the Ethics Committee of Suzhou Wujiang District Children's Hospital (approval number: 2022010). First, hematoxylin-eosin (H&E) staining was used to evaluate the effects of SA on IRI, and renal damage was scored. Serum creatinine (SCr), blood urea nitrogen (BUN) and cystatin C (Cystatin C) were analyzed. The effect of sappanone A on the apoptosis of renal tubular epithelial cells induced by IRI was analyzed by TUNEL staining. Protein expression levels of p-JNK/JNK, p-ERK/ERK, Bcl2, Bax and cleaved-caspase 3 in renal tissues were detected by Western blot. Finally, H&E staining, serological analysis, TUNEL staining and Western blot were used to determine whether JNK activator anisomycin could reverse the effect of SA on IRI in rats. The results showed SA significantly reduced the renal tubule injury caused by ischemia-reperfusion, and decreased the level of SCr, BUN and Cys C in serum. TUNEL staining showed that SA significantly reduced the apoptosis of renal tubular epithelial cells induced by IRI. Western blot analysis of kidney tissue showed that SA significantly promoted the expression of apoptosis inhibiting protein Bcl2 and inhibited the expression of apoptosis-promoting proteins Bax and cleaved-caspase 3. Further analysis elucidated that SA did not affect the phosphorylation of ERK but decreased the phosphorylation of JNK. Finally, H&E staining, serological analysis, TUNEL staining and Western blot confirmed that JNK activator anisomycin could reverse the alleviating effect of SA on IRI in rats. The above findings suggest that SA could alleviate IRI in rats by inhibiting JNK phosphorylation.

In order to solve the problem of resistance of Pseudomonas aeruginosa to multiple antibiotics, it is an effective way to find inhibitors of P.aeruginosa efflux pump. In this study, 15 new ornithine peptidomimetic derivatives were designed and synthesized by changing the side chain structure of natural amino acids with PAβN, a dipeptide efflux pump inhibitor, and their synergic activity with aztreonam, a monocyclic β-lactam antibiotic, against P.aeruginosa was evaluated. Among them, the representative compound 12b not only enhanced the antibacterial activity of β-lactam antibiotics aztreonam, ceftazidime and meropenem, but also significantly enhanced the antibacterial action of macrolide antibiotics clarithromycin, showing a broad-spectrum synergic sensitization effect. In addition, compound 12b also has a good safety. Preliminary mechanisms suggest that 12b works by directly targeting the efflux transporter MexB. These results provide a new lead compound for the development of a new class of efflux pump inhibitors against P.aeruginosa.

Kidney ischemia reperfusion injury (IRI) is a leading cause of acute kidney injury (AKI) with a poor prognosis and high mortality rate. Recent studies have reported that chrysophanol may have a renal protective effect, but its specific impact and mechanism on IRI remain unclear. This study aimed to explore the effects and mechanisms of chrysophanol on AKI induced by IRI. By utilizing a unilateral kidney IRI mouse model, histopathological changes in the kidney, serum levels of creatinine and urea nitrogen, and protein expressions of apoptosis and mitophagy in kidney tissue were examined. Additionally, a hypoxia/reoxygenation (H/R) model of human kidney-2 (HK-2) cells was established to measure mitochondrial membrane potential levels and reactive oxygen species (ROS). Functional enrichment analysis was performed to screen relevant targets of chrysophanol and AKI, and to verify key targets and pathways. The animal experiments conducted in this study were ethically approved by the Experimental Animal Ethics Committee of Peking University (No. LA2021503). The findings indicate that the IRI group exhibited elevated levels of creatinine and urea nitrogen in serum, significant renal tissue damage, and increased expression of renal injury markers (KIM1), apoptosis-related proteins (cleaved-caspase 3, caspase 3, cytochrome C), and mitochondrial autophagy protein (PINK1) compared to the sham surgery group. Chrysophanol treatment ameliorated the aforementioned pathological changes in a dose-dependent manner in an IRI model. Additionally, it exhibited significant improvements in mitochondrial membrane potential and inhibition of ROS production in HK-2 cells subjected to H/R conditions. Through network pharmacological analysis, HSP90AA1 and PIK3R1 were identified as key targets primarily enriched in the phosphoinositide 3 kinase/protein kinase B (PI3K/Akt) pathway. Real-time quantitative PCR (qPCR) validation confirmed that chrysophanol significantly decreased HSP90AA1 and PIK3R1 mRNA levels in HK-2 cells under H/R conditions, while also enhancing the protein expressions of p-PI3K, PI3K, p-Akt, and Akt. In conclusion, chrysophanol has the potential to enhance AKI by selectively modulating HSP90AA1 and PIK3R1, activating the PI3K/Akt pathway, decreasing apoptosis, regulating mitochondrial autophagy, enhancing mitochondrial membrane potential, and suppressing ROS production. These findings suggest that chrysophanol could serve as a promising therapeutic option for the treatment of AKI.

Pentacyclic triterpenoids are a class of widespread natural compounds containing six isoprene structures with a wide range of pharmacological activities, including antibacterial, anti-inflammatory, antiviral, antitumor, immune regulation, etc. The structural modifications of pentacyclic triterpenoid natural products and the drug development have always been a hot research topic. This article reviews the recent progresses in the structural modifications, pharmacological effects, and clinical studies of different kinds of pentacyclic triterpenoid natural products.

Kynurenine 3-monooxygenase (KMO) is a key rate-limiting enzyme in the downstream catabolism of kynurenine pathway (KP). Under the catalysis of KMO, the intermediate product kynurenine is metabolized into various active metabolites, including 3-hydroxykynurenine (3-HK), quinolinic acid (QA) and nicotinamide adenine dinucleotide (NAD⁺). More and more studies have shown that abnormal KMO expression activity mediates KP metabolic disorders, and is involved in the occurrence and development of nervous system diseases, autoimmune diseases, infectious diseases and tumors, suggesting that KMO can be used as a potential and effective drug therapeutic target. This article focuses on the role of KMO in the pathological mechanism of various diseases, and summarizes the existing KMO inhibitors to provide methods and ideas for targeted KMO therapy.

GNPS-based mass spectrum-molecular networks is an effective strategy for rapidly identifying known natural products and discovering novel structures. The chemical diversity of azaphilones from the fermentation extracts of Talaromyces sp. HK1-18 was studied by molecular network technique. Three linear tricyclic azaphilones, sequoiamonacins A-C (2a, 2b, 1), were isolated by silica gel column chromatography and high performance liquid chromatography from the extracts of the fungal strain of HK1-18, and their structures were identified by nuclear magnetic resonance and high-resolution mass spectrometry. Guided by the mass spectra of sequoiamonacins A-C (2a, 2b, 1), the cluster of sequoiamonacinoid analogues was discovered from the full molecular networking of HK1-18. By analyzing the MS/MS fragments of each parent ion in this cluster, 7 azaphilones (3-9) included 6 new ones (4-9) were predicted successfully. Then the MS/MS cracking regularity of this type of azaphilones was revealed. Compound 1 showed anti-inflammatory activity, which can inhibit the production of interleukin-1α (IL-1α) in lipopolysaccharide (LPS)-induced mouse macrophage RAW264.7, with an inhibitory rate of 29% at the concentration of 12.5 μg·mL^-1.