ObjectiveTo analyze the long-term trend of incidence and mortality of type 2 diabetic kidney disease （DKD） in China from 1990 to 2021. MethodsThe Joinpoint regression model was used to analyze the average annual percentage change （AAPC） of standardized incidence rate and standardized mortality rate， and the age-period-cohort （APC） model was constructed to analyze the longitudinal age change， period and cohort effect risk ratio （RR）. ResultsFrom 1990 to 2021， the standardized incidence rate of type 2 DKD in males and females showed an overall upward trend， with AAPC of 0.08% and 0.36%， respectively. The age-standardized mortality rate of the total population and female showed a downward trend， with AAPC of -0.61% and -1.03%， respectively. However， there was no significant difference in males. APC model showed that the age effect existed： the peak age was 75-79 years old， the mortality rate of females increased， and the mortality rate of males decreased after 80-84 years old. For the effect of time period， the risk of type 2 DKD incidence in females in 2017—2021 was 1.05 times that in 2002—2006， and the risk of death in males and females in 2017—2021 was 0.84 and 0.71 times that in 2002—2006， respectively. For cohort effects， the highest risk of disease was seen in men and women born in 1967—1971， and the highest risk of death was seen in men born in 1952—1956 and women born in 1912—1916. ConclusionFrom 1990 to 2021， the standardized incidence rate of type 2 DKD in China shows an upward trend， and the standardized mortality rate shows a downward trend. It is necessary to strengthen the health behavior publicity and education of type 2 DKD， and actively carry out early screening to reduce the disease burden.

Metabolic dysfunction-associated fatty liver disease （MAFLD）， as one of the most common chronic liver diseases in the world， poses a severe challenge to precision diagnosis and treatment due to its complex pathogenesis and highly heterogeneous disease progression. Existing clinical classification systems cannot meet the needs for comprehensively analyzing the complexity of the disease and the heterogeneity of its adverse outcomes. In recent years， data-driven prognostic risk classification methods have gradually emerged， optimizing the ability for predicting adverse outcomes and enhancing the accuracy of identifying different endpoint outcomes. However， such paradigm of “classify first， associate outcomes later” suffers from a “black-box” nature， and there are various indicators for classification， leading to limited stability and generalizability in clinical application. Future research needs to integrate or establish large-scale population cohorts， develop outcome-oriented prognostic risk classification models， incorporate dynamic data， refine classification algorithms， and validate their generalizability across multiple populations， thereby providing reliable support for the precision diagnosis and treatment of MAFLD.

ObjectiveTo compare the differences between the ultrafiltration method and the dextran gel filtration method during the purification of Tfn-modified PCL micelles by using purification efficiency and micelle purity as indicators. MethodsCoumarin-6 （C6） was used as a fluorescent probe and was loaded into HOOC-PEG-PCL to form PCL micelles by the film-dispersion method. Tfn was then conjugated to the surface of PCL micelles via an amidation reaction， resulting in two types of micelles： Tfn/PCL_H and Tfn/PCL_L. The pharmaceutical properties of the two types of micelles were characterized. The micelles were then purified through ultrafiltration method and dextran gel method respectively， and the efficiency of the two methods， along with the purity of the final micelles， was compared. The density of Tfn on the surface of PCL micelles was also calculated. ResultsThe hydrated diameter of PCL micelles was approximately 73 nm， and the C6 loading efficiency was around 0.046%. The size increased to 134 nm and 158 nm for Tfn/PCL_L and Tfn/PCL_H， respectively. The micelle population was monodisperse. The purification results showed that， for the ultrafiltration method， after two and one rounds of purification， the Tfn/C6 ratio stabilized at 23.6 and 3.4 for Tfn/PCL_H and Tfn/PCL_L， respectively. For the dextran gel filtration method， the Tfn/C6 ratio reached 23.7 for the Tfn/PCL_H group after two rounds of purification. However， for the Tfn/PCL_L group， the Tfn/C6 ratio increased during four rounds of dextran gel purification， and a significant difference （P = 0.042 4） was observed between the first and last filtrations. The density of Tfn in the final micelles were calculated. For the ultrafiltration method， the Tfn density of Tfn/PCL_H and Tfn/PCL_L were 94.9% and 13.8%， respectively. For the dextran gel filtration method， the density of the two micelles were 95.6% and 14.4%， respectively. For Tfn/PCL_L group， the density results revealing a statistically significant difference （P=0.000 2）. ConclusionThe purification efficiency of the two methods is comparable. However， the purity of the final micelles shows a significant difference， with the dextran gel filtration method resulting in higher purity， particularly for the Tfn/PCL_L micelles.

ObjectiveAbnormal lipids in neurons can cause the accumulation of α-synuclein（α-syn）. This study aimed to explore the mechanism of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis extract （ASH） in treating Parkinson's disease （PD） mice using lipidomics combined with network pharmacology. MethodsMice were divided into the blank group， model group and ASH （45.5 mg·kg^-1） group. Motor ability was evaluated by pole climbing time and autonomous activity count； The oxidative stress indicators were detected by enzyme-linked immunosorbent assay （ELISA）. Lipid biomarkers in brain tissues were screened and identified by ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）， and metabolic pathway analysis was conducted. The key targets of ASH for PD treatment were explored using network pharmacology. The Kyoto Encyclopedia of Genes and Genomes （KEGG） database was used for pathway enrichment analysis， and the "compound-reaction-enzyme-gene" network was constructed using the MetScape plugin. The protein expression levels of glutathione S-transferase P1 （GSTP1）， glutathione S-transferase Mu 2 （GSTM2）， prostaglandin peroxide synthase 1 （PTGS1）， prostaglandin peroxide synthase 2 （PTGS2）， and prostaglandin E synthase （PTGES） were validated by Western blot. ResultsCompared with the blank group， the model group showed significantly prolonged pole climbing time and reduced autonomous activity count （P<0.01）. Compared with the model group， the ASH group demonstrated significantly faster pole climbing and increased autonomous activity count （P<0.01）. The model group exhibited significantly decreased superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） levels， and increased malondialdehyde （MDA） level in brain tissues compared with the blank group （P<0.01）. The ASH group showed increased SOD and GSH-Px levels and decreased MDA level compared with the model group （P<0.05， P<0.01）. Lipidomics analysis identified 10 differential metabolites and 8 differential metabolic pathways. Network pharmacological analysis revealed 213 intersection targets between ASH components and PD， with KEGG enrichment involving the sphingolipid signaling pathway， lipid arteriosclerosis， phosphoinositide 3-kinase/protein kinase B（PI3K/Akt） signaling pathway， mitogen-activated protein kinase（MAPK） signaling pathway， and hypoxia inducible factor-1（HIF-1） signaling pathway. Integrated lipidomics and network pharmacology analysis highlighted the central role of the arachidonic acid metabolic pathway. The Western blot results showed that ASH effectively up-regulated GSTP1， GSTM2， and PTGS1 protein expression， and down-regulated PTGS2 and PTGES protein expression. ConclusionASH can ameliorate behavioral deficits， exert antioxidant effects， regulate lipid differential metabolites and the arachidonic acid metabolic pathway， thereby exerting therapeutic effects in PD model mice.

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.

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 relationship between the expression of serum microRNA(miR)-488 and miR-184 and the occurrence of preeclampsia in patients with gestational hypertension.Methods A to-tal of 128 patients with gestational hypertension admitted to a hospital from June 2022 to March 2023 were se-lected and divided into the simple gestational hypertension group(55 cases)and the combined with pre-eclampsia group(73 cases)according to whether they developed preeclampsia.Another 46 healthy pregnant women in the same period were selected as the control group.Real-time fluorescence quantitative polymerase chain reaction was used to detect miR-488 and miR-184 expression in each group,and multiple Logistic regres-sion was used to analyze the influencing factors of the occurrence of preeclampsia in patients with gestational hypertension.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of each influencing factor in the occurrence of preeclampsia in patients with gestational hypertension,and Pearson cor-relation was used to analyze the correlation between serum miR-488,miR-184 and placental growth factor.Re-sults Compared with the control group,the expressions of serum miR-488 and miR-184 were increased in the simple pregnancy hypertension group and the combined with preeclampsia group,and the differences were sta-tistically significant(P<0.05).Compared with mild preeclampsia group,serum miR-488 and miR-184 ex-pressions were increased in severe preeclampsia group,and the difference was statistically significant(P<0.05).There were significant differences in gravida and parity,systolic blood pressure,diastolic blood pres-sure,urinary protein quantity and placental growth factor levels between the combined with preeclampsia group and the simple pregnancy hypertension group(P<0.05).Multiple Logistic regression analysis showed that gravida and parity,systolic blood pressure,diastolic blood pressure,urinary protein quantity,miR-488 and miR-184 were risk factors for preeclampsia in patients with gestational hypertension(P<0.05),and placental growth factor was protective factor for preeclampsia in patients with gestational hypertension(P<0.05).ROC curve analysis showed that the prediction efficiency of miR-488+miR-184 was significantly better than that of serum miR-488(Z=2.530,P=0.011)and miR-184(Z=2.437,P=0.015)alone.Serum miR-488 and miR-184 were negatively correlated with placental growth factor(r=-4.361,-4.288,P<0.05).Con-clusion Serum miR-488 and miR-184 have predictive value in the occurrence of preeclampsia in patients with gestational hypertension.

Objective:To observe the effect of governor vessel(GV)electroacupuncture on potassium chloride cotransport-er 2(KCC2)and γ-Aminobutyric acid type a receptor(GABAAR α 1)and to explore the possible mecha-nism of GV electroacupuncture in improving post-stroke limb spasticity(PSLS).Method:45 male SD rats of SPF grade were randomly selected into the blank group and the sham operation group,with 9 rats in each group.The remaining rats were used to make the middle cerebral artery occlusion model.On the 3rd day after the completion of modeling,the rats with limb spasticity were chosen and random-ly divided into the model group,the electroacupuncture group,and the baclofen group,with 9 rats in each group,following a series of behavioral tests.Acupuncture was provided to the electroacupuncture group at"Da-zhui","Jizhong"and"Houhui",whereas the baclofen group received a baclofen solution by gavage for 7 days.Rats in each group were observed for the changes in nerve function by the symptom score of nerve function defect,and the changes in muscle tension level were evaluated by the modified Ashworth grading score and EMG signal value change.KCC2 and GABAARα1 mRNA and protein in the cerebral cortex of rats were de-tected by qPCR and Western Blot,and changes in chloride ion content were detected with a colorimetric kit.Result:The neurological function defect score,the modified Ashworth grading score,the water content of brain tissue,the volume of cerebral infarction,and the expression of chloride ion content in the model group were all significantly increased compared with the sham operation group at the same time point(P＜0.01),while the EMG signal value,cerebral cortex KCC2 and GABAAR α 1 mRNA and protein were significantly decreased(P＜0.01).Compared with the model group,the nerve function defect score,the modified Ashworth grading score,the water content of brain tissue,the volume of cerebral infarction and the expression of chlo-ride ion content in the electroacupuncture group and the baclofen group were all decreased(P＜0.01),while the EMG signal value and KCC2,with GABAARα1 mRNA and protein expression were all increased(P＜0.01,P＜0.05,respectively);There was no significant difference in all indexes between the GV electroacupuncture group and the baclofen group(P＞0.05).Conclusion:GV electroacupuncture can increase the expression of KCC2,GABAARα1 mRNA,and protein in the cerebral cortex of rats,in order to exert the inhibitory effect of γ-aminobutyric acid,thus improving the symptoms of PSLS.

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