Objective: Scutellarin in Shuganning injection has potential allergenicity and is prone to causing anaphylactoid reactions. There is an urgent need to establish a simple and rapid chromatographic quantitative method to achieve the limit control of scutellarin in Shuganning injection.
Methods: For scutellarin, HPLC method was used in the quantitative analysis using a Shim-pack GIS C₁₈ (2.1 mm×150 mm, 5 μm) column. The mobile phase was a blend of 0.1% trifluoroacetic acid in water and acetonitrile, and gradient elution progress was applied for the whole process. The flow rate was 0.3 mL·min^-1 and the column temperature was maintained at 37 ℃. The detecting wavelength was 320 nm.
Results: The linear regression equation was y=51 456x-27 352, the correlation coefficient r=0.999 8, namely the calibration curve was linear in the range of 1.02-102.00 μg·mL^-1; the average recovery was 103.76%; the detection and quantification limit for determining scutellarin was 0.16 μg·mL^-1 and 0.57 μg·mL^-1, respectively; the injection sample was stable at room temperature for 48 h.
Conclusion: This method is easy to operate, reproducible and has high accuracy, thus can be used for quality control of Shuganning injection.

Hyperthermic intrathoracic chemotherapy (HITHOC) is a localized treatment that combines hyperthermia with medications such as chemotherapy, anti-angiogenic agents, targeted therapy, or immunotherapeutic agents. It has advanced clinical progress for conditions such as malignant pleural mesothelioma, pleural metastasis of lung cancer, and pleural dissemination of thymic tumors. Systematic analyses indicate that as an adjuvant therapy to cytoreductive surgery, HITHOC improves patient survival and effectively controls pleural effusion and pleural metastasis. It exerts antitumor effects by enhancing drug penetration through thermal effects, inducing apoptosis, and modulating the immune microenvironment. HITHOC combined with systemic therapies (e.g., chemotherapy, targeted therapy, and immunotherapy) demonstrates synergistic potential, but its role as a local treatment should be clearly defined as adjunctive within the comprehensive management of advanced-stage patients. Further research is needed to clarify its indications, standardized protocols, and long-term efficacy and to explore optimized regimens and personalized combination treatment strategies. This review delves into the key bottlenecks hindering the development of hyperthermia therapy, such as the lack of high-level evidence-based medicine and the absence of standardized diagnostic and efficacy evaluation criteria, and proposes potential solutions.

The JAK2 V617F mutation is a key molecular event in myeloproliferative neoplasms and is strongly associated with an increased risk of thrombosis. This review aims to summarize the underlying mechanisms, clinical manifestations, and management strategies for thrombosis driven by the JAK2 V617F mutation. Relevant recent studies are analyzed to evaluate the effect of the mutation on molecular signaling, hematopoietic cell function, inflammatory activation, endothelial injury, and coagulation imbalance. Findings indicate that the JAK2 V617F mutation induces the persistent activation of the JAK-STAT pathway, leading to erythroid clonal expansion, platelet hyperreactivity, and enhanced neutrophil chemotaxis, along with excessive inflammatory cytokine release, NET formation, and platelet-leukocyte aggregation. These processes collectively promote endothelial dysfunction and hypercoagulation, ultimately contributing to arterial and venous thrombosis. Clinically, affected patients may present with coronary artery events deep vein thrombosis, and portal vein thrombosis, often accompanied by high recurrence rates and poor outcomes. In conclusion, the JAK2 V617F mutation drives thrombosis via multiple intertwined mechanisms, suggesting that future therapeutic strategies should integrate anticoagulation, anti-inflammation therapy, NET inhibition, and targeted JAK-STAT pathway blockade to improve patient survival.

One of the technical bottlenecks limiting the high yield of 1,4-butanediamine is the insufficient tolerance of strains to 1,4-butanediamine. Enhancing the tolerance of strains to 1,4-butanediamine is therefore a primary challenge that needs to be addressed for the construction of strains with high yields of 1,4-butanediamine. Staphylococcus pasteuri 326180 exhibits exceptional tolerance to high-concentration 1,4-butanediamine, serving as both an ideal model for studying the mechanism underlying the 1,4-butanediamine tolerance and a novel host for constructing strains capable of efficiently producing 1,4-butanediamine. However, for both the research on the tolerance mechanism and the modification of chassis strains, gene editing of S. pasteuri needs to be carried out at the molecular level. The research objective of this paper is to establish a genetic manipulation system for S. pasteuri, laying foundation for subsequent studies on tolerance mechanism and the modification of chassis strains. This study systematically optimized the electroporation conditions, including key parameters such as the growth phase of cells, electric field strength, electroporation buffer, and recovery medium, successfully establishing an electroporation method for S. pasteuri. Additionally, we constructed the gene editing plasmid pCpfOA by replacing the resistance expression cassette, optimized the selection markers for gene editing, and finally established a CRISPR/Cpf1-based gene editing technology for S. pasteuri, achieving an editing efficiency of 90%. The genetic manipulation system of S. pasteuri established in this study provides technical support for research into the tolerance mechanism of this bacterium and the genetic modification of chassis strains.
Staphylococcus/drug effects* ; Gene Editing/methods* ; Electroporation/methods* ; Plasmids/genetics* ; CRISPR-Cas Systems ; Genetic Engineering/methods*

Objective To investigate the value of serum lipoprotein-associated phospholipase A2(Lp-PLA2)for predicting high-risk coronary plaques in elderly males.Methods A retrospective study was conducted on 46 elderly males aged ≥60 years undergoing health check-ups and coro-nary computed tomography angiography in our hospital between May and July 2024.Their general clinical data were collected.Artificial intelligence software was used to analyze coronary calcium scores and plaque characteristics.The participants were divided into a high-risk plaque group(n=15)and a non-high-risk plaque group(n=31).The differences were compared between the two groups.Multivariate logistic regression analysis was used to identify the influencing factors for high-risk coronary plaques.ROC curve was plotted to determine the predictive value of serum Lp-PLA2 for high-risk plaques,and its AUC value was calculated.Results The high-risk plaque group had significantly larger proportions of smoking history and hyperlipidemia,and higher level of homocysteine and Lp-PLA2 than the non-high-risk plaque group(P＜0.05,P＜0.01).Multiva-riate logistic regression analysis indicated that Lp-PLA2 was an independent risk factor for high-risk coronary plaques(HR=1.030,95％CI:1.008-1.053,P＜0.05).ROC curve analysis revealed that the AUC value of Lp-PLA2 in predicting high-risk coronary plaques was 0.833(95％CI:0.694-0.927,P＜0.01),with a sensitivity of 93.3％,a specificity of 71.0％,a positive predictive value of 62.5％,and a negative predictive value of 100％.Conclusion Serum Lp-PLA2 is of signif-icant value in predicting high-risk coronary plaques in elderly men.

OBJECTIVE To investigate a suspected outbreak of carbapenem-resistant Klebsiella pneumoniae(CRKP)infection in the intensive care unit of a traditional Chinese medicine hospital,identify the source of infec-tion and transmission routes,and provide a basis for prevention and control of CRKP infection.METHODS Epide-miological investigations were conducted on five patients with CRKP infections or colonization who were identi-fied in Jul.2024 at Suiyang County Hospital of Traditional Chinese Medicine.Samples were collected from pa-tients,the ward environments,and hand surfaces to detect CRKP.Fourteen CRKP isolates were selected for car-bapenemase gene testing,and homology analysis was performed by enterobacterial repetitive intergenic consensus polymerase chain reaction(ERIC-PCR)and multilocus sequence typing(MLST).RESULTS The median age of the five cases was 73 years,and all had undergone multiple invasive procedures.Environmental monitoring showed a CRKP positive rate of 26.35％,with CRKP isolates detected on the hands of healthcare workers,surfaces in the wards and medical equipment surfaces.Genetic analysis showed that all 14 CRKP strains carried the KPC resist-ance gene;except for case 1,other strains carried the VIM gene.MLST identified CRKP of all strains as sequence type 48(ST48);while ERIC-PCR revealed two distinct genotypes:genotype A for case 1 and genotype B for the other cases and environmental isolates.After strengthening patient isolation and group treatment,strictly cleaning and disinfecting the ward environments and medical equipment,and strictly implementing hand hygiene,the infec-tion was effectively controlled.CONCLUSIONS Inadequate disinfection of the ward environments and medical e-quipment and poor compliance with hand hygiene are the main contributors to the suspected CRKP outbreak.Ho-mology analysis suggests the existence of two independent transmission chains.Timely identification and manage-ment of the infection sources,interruption of transmission routes,protection of susceptible individuals and imple-mentation of comprehensive infection control measures are essential for effective outbreak control.

Objective To investigate the relationship between four metabolic indices reflecting insulin resistance(IR)and the risk of diabetic kidney disease(DKD)in patients with type 2 diabetes mellitus(T2DM).Methods 157 T2DM patients with DKD(DKD group)and 198 T2DM patients(T2DM group)were included in our study.The general data and clinical indicators were collected.Triglyceride/high density lipoprotein cholesterol ratio(TG/HDL-C),triglyceride glucose(TyG)index,triglyceride glucose-body mass index(TyG-BMI)and metabolic score for IR(METS-IR)were calculated.Results Compared with T2DM group,TG/HDL-C,TyG index,TyG-BMI and METS-IR in DKD group were significantly increased(P<0.05).Logistic regression analysis showed that TyG index(OR=8.303,95%CI 1.787～38.573,P=0.007)was an independent risk factor for DKD.The risk of DKD in the fourth quantile of TyG index group was 5.652 times than the first quantile group(P<0.05).ROC curve analysis showed that when the cutoff value of TyG index was 9.03,its sensitivity and specificity for predicting DKD were 56.7%and 71.7%,respectively.Conclusions Among the four simple IR indicators,TyG index is an independent risk factor for the occurrence of DKD in T2DM patients.Increased TyG index indicates increased UACR level and high risk of DKD.

Objective To investigate the mechanism by which Guijianyu ameliorates podocyte injury in a mouse model of diabetic kidney disease(DKD)induced by advanced glycation endproducts(AGEs).Methods Sixty db/db mouse models of DKD were randomized equally into 5 groups for treatment with saline,Guijianyu extract at 3 doses or irbesartan for 12 weeks,and the changes in renal pathology and structure were observed using transmission electron microscopy,and the expressions of related genes and key proteins were detected using RT-qPCR and immunohistochemistry.In cultured MPC-5 cells incubated with 50 mg/L AGEs-BSA for 24 h,the effect of different concentrations of Guijianyu extract on cell viability was examined with CCK-8 assay;Western blotting was performed to detect the protein expressions of RAGE,VEGFA,TNF-α,NF-κB(p65),IL-6 and caspase-3,and the mRNA expressions of RAGE,NF-κB(p65),VEGFA and IL-6 were detected with RT-qPCR.Results In mouse models of DKD,treatment with high-dose Guijianyu extract significantly reduced renal expressions of RAGE,VEGFA,NF-κB(p65),and IL-6 proteins and the mRNA expressions of RAGE,NF-κB,and IL-6.In MPC-5 cells,exposure to AGEs significantly reduced cell viability and increased the protein expressions of RAGE,NF-κB(p65),VEGFA,TNF-α,IL-6 and caspase-3(P<0.05)and mRNA expressions of RAGE,NF-κB(p65),VEGFA,and IL-6.Treatment with Guijianyu extract obviously improved cell viability and reduced the expressions of RAGE,NF-κB(p65),VEGFA,TNF-α,IL-6,and caspase-3.Furthermore,Guijianyu extract effectively reversed RAGE agonist-induced elevation of protein expressions of RAGE,VEGFA,TNF-α,IL-6,and caspase-3 and mRNA expressions of RAGE,NF-κB(p65),IL-6,and VEGFA in MPC-5 cells.Conclusion Guijianyu extract ameliorates AGEs-induced mouse renal podocyte injury in DKD by inhibiting the activation of AGEs-RAGE signaling pathway and reducing the expressions of pro-inflammatory cytokines and vascular endothelial growth factors.

Metastatic dissemination is the major cause of death from breast-cancer (BC). Fusobacterium nucleatum (F.n) is widely enriched in BC and has recently been identified as one of the high-risk factors for promoting BC metastasis. Here, with an experimental model, we demonstrated that intratumoral F.n induced BC aggressiveness by transcriptionally activating Epithelial-mesenchymal transition-associated genes. Therefore, the F.n may be a potential target to prevent metastasis. Given the fact that cancer-associated fibroblasts (CAFs) are abundant in BC and located near blood vessels, we report an optogenetic system that drives CAF to in situ produce human antibacterial peptide LL37, with the characteristics of biosafety and freely intercellular trafficking, for depleting intratumoral F.n, leading to a 72.1% reduction in lung metastatic nodules number without affecting the balance of the systemic flora. Notably, mild photothermal treatment was found that could normalize CAF, contributing to synergistically inhibiting BC metastasis. In addition, the system can also simultaneously encode a gene of TNF-related apoptosis-inducing ligand to suppress the primary tumor. Together, our study highlights the potential of local elimination of tumor pathogenic bacteria to prevent BC metastasis.

OBJECTIVES: To investigate the mechanism by which Guijianyu ameliorates podocyte injury in a mouse model of diabetic kidney disease (DKD) induced by advanced glycation endproducts (AGEs). METHODS: Sixty db/db mouse models of DKD were randomized equally into 5 groups for treatment with saline, Guijianyu extract at 3 doses or irbesartan for 12 weeks, and the changes in renal pathology and structure were observed using transmission electron microscopy, and the expressions of related genes and key proteins were detected using RT-qPCR and immunohistochemistry. In cultured MPC-5 cells incubated with 50 mg/L AGEs-BSA for 24 h, the effect of different concentrations of Guijianyu extract on cell viability was examined with CCK-8 assay; Western blotting was performed to detect the protein expressions of RAGE, VEGFA, TNF-α, NF-κB(p65), IL-6 and caspase-3, and the mRNA expressions of RAGE, NF-κB (p65), VEGFA and IL-6 were detected with RT-qPCR. RESULTS: In mouse models of DKD, treatment with high-dose Guijianyu extract significantly reduced renal expressions of RAGE, VEGFA, NF-κB(p65), and IL-6 proteins and the mRNA expressions of RAGE, NF-κB, and IL-6. In MPC-5 cells, exposure to AGEs significantly reduced cell viability and increased the protein expressions of RAGE, NF‑κB (p65), VEGFA, TNF-α, IL-6 and caspase-3 (P<0.05) and mRNA expressions of RAGE, NF-κB (p65), VEGFA, and IL-6. Treatment with Guijianyu extract obviously improved cell viability and reduced the expressions of RAGE, NF-κB(p65), VEGFA, TNF-α, IL-6, and caspase-3. Furthermore, Guijianyu extract effectively reversed RAGE agonist-induced elevation of protein expressions of RAGE, VEGFA, TNF-α, IL-6, and caspase-3 and mRNA expressions of RAGE, NF-κB (p65), IL-6, and VEGFA in MPC-5 cells. CONCLUSIONS Guijianyu extract ameliorates AGEs-induced mouse renal podocyte injury in DKD by inhibiting the activation of AGEs-RAGE signaling pathway and reducing the expressions of pro-inflammatory cytokines and vascular endothelial growth factors.
Animals ; Glycation End Products, Advanced ; Drugs, Chinese Herbal/pharmacology* ; Mice ; Signal Transduction/drug effects* ; Podocytes/pathology* ; Diabetic Nephropathies/drug therapy* ; Receptor for Advanced Glycation End Products ; Vascular Endothelial Growth Factor A/metabolism* ; Interleukin-6/metabolism* ; Male