OBJECTIVE To study the protective effect and potential mechanism of chikusetsu saponin Ⅳ a （chsⅣ） on renal function in diabetic nephropathy （DN） model rats. METHODS DN rat model was established by high-fat diet combined with streptozotocin injection. Thirty-six model rats were randomly divided into model group （i.g. administration of normal saline， high-fat diet）， chsⅣ low-dose and high-dose groups （i.g. administration of 90， 180 mg/kg chsⅣ， high-fat diet）， with 12 rats in each group. Additionally， 10 normal rats were set as the control group （i.g. administration of normal saline， regular diet）. From the 5th to the 12th week after streptozotocin injection， they were given intragastric administration of relevant drug or normal saline， once a day. After the last medication， the levels of fasting blood glucose， fasting insulin， blood urea nitrogen， serum creatinine and urine protein as well as the levels of reduced glutathione （GSH）， superoxide dismutase （SOD） and malondialdehyde （MDA） in renal tissues were measured. Additionally， the insulin resistance index was calculated. Hematoxylin-eosin， periodic acid-Schiff， and Masson staining techniques were employed to examine the histopathological alterations in the renal tissue. The expressions of Notch signaling pathway-related proteins in renal tissue were detected by immunohistochemical staining and Western blot methods. RESULTS Compared with model group， the histomorphological of renal tissues in the chsⅣ low- and high-dose groups were significantly improved， with significant decreases in renal histological scores， mesangial expansion index， and glomerulosclerosis scores （ P ＜0.05）； the levels of fasting blood glucose， fasting insulin， blood urea nitrogen， serum creatinine， urine protein and homeostasis model assessment for insulin resistance， as well as MDA content， the expression levels of Notch1， Notch intracellular domain， hairy and enhancer of Split 1 and Delta-like protein 1 in renal tissue were all significantly decreased （ P ＜0.05）. The levels of GSH and SOD in renal tissue were significantly elevated （ P ＜0.05）. Moreover， the improvement in these indicators was significantly more pronounced in the chsⅣ high-dose group compared to the chsⅣ low-dose group （ P ＜0.05）. CONCLUSIONS ChsⅣ can ameliorate renal pathological damage and functional impairment in DN rats. Its underlying mechanisms include restoration of glucose homeostasis and insulin sensitivity， attenuation of renal oxidative stress， and suppression of aberrant Notch signaling pathway activation.

Autosomal dominant polycystic kidney disease （ADPKD） is a systemic hereditary renal disorder and can affect multiple organs， and congenital hepatic fibrosis is one of the manifestations of liver involvement and is an important complication of ADPKD. Symptomatic management is currently the main treatment method for this disease， and disease-specific drugs such as tolvaptan have limited indications and cannot correct the underlying genetic defect. This article reports a case of ADPKD with congenital hepatic fibrosis， and sirolimus was identified as the individualized treatment regimen based on peripheral blood functional genomic analysis and drug sensitivity prediction platform. The patient achieved significant improvements in symptoms and quality of life after treatment， with a stable kidney volume. This case shows that functional genomics has a potential value in guiding individualized treatment of rare genetic disorders， which provides new treatment ideas and practice paths for similar patients.

OBJECTIVE To provide an experimental basis for quality control， clinical application and development of hepatoprotective drugs derived from Thesium chinense Turcz.METHODS Network pharmacology was adopted to predict and screen potential hepatoprotective constituents and their action targets of T. chinense Turcz. A liver-injured model of transgenic zebrafish Tg （ fabp10a ： dsRed ） with fluorescent-labeled liver was induced by 4 mmol/L isoniazid. The relative liver fluorescence area and relative fluorescence intensity of zebrafish larvae， as well as the levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， glutathione （GSH） and interleukin-6 （IL-6） were taken as indexes to evaluate the hepatoprotective activity of potential active constituents. High-performance liquid chromatography was used to determine the contents of hepatoprotective active constituents in 17 batches of T. chinense Turcz. medicinal materials from different producing areas.RESULTS Four potential hepatoprotective constituents including kaempferol-3- O -glucorhamnoside （KOG）， kaempferol-3，7-di- O -glucoside （KDG）， buddleoside （BUD） and apigenin-5- O -neohesperidoside （AON） were screened out via network pharmacology， and their core targets involved mitogen-activated protein kinase 1， nuclear factor-κB subunit 1， etc. Pharmacodynamic verification results showed that 25 μmol/L KOG， BUD， AON and 50 μmol/L KDG could significantly increase the relative liver fluorescence area and relative fluorescence intensity of liver-injured model larvae （ P ＜0.05）， reduce the levels of AST， ALT and IL-6 （ P ＜0.05）， and elevate the GSH level （ P ＜0.05）. Content determination results indicated that KOG could be detected in all 17 batches of T. chinense Turcz. medicinal materials， with contents ranging from 0.097 0 to 8.082 0 mg/g；KDG， AON and BUD were not detected in partial batches of samples， and their contents in detectable batches were 0.019 3-0.123 2 mg/g， 0.018 4-0.109 1 mg/g and 0.385 1-1.189 3 mg/g， respectively.CONCLUSIONS KOG， KDG， AON and BUD in T.chinense Turcz. may be its potential hepatoprotective active constituents. Among them， KOG can be detected in T.chinense Turcz. medicinal materials from different producing areas with relatively high content， which can be used as the main marker constituent for evaluating the hepatoprotective effect of T.chinense Turcz.

Large vessel occlusion due to intracranial atherosclerotic disease (ICAD-LVO) is one of the types of ischemic stroke with poor outcome. This article summarizes the relevant studies of ICAD-LVO in recent years, and reviews the imaging evaluation and treatment regimens to provide reference for clinical practice.

Objective:To investigate the molecular mechanisms of bexarotene in treating double hit lymphoma(DHL)based on Weighted Gene Co-expression Network Analysis(WGCNA),thereby providing potential targets and experimental evidence for DHL treatment.Methods:The gene expression datasets GSE44164 and GSE43677 were downloaded from the Gene Expression Omnibus(GEO)database,and differentially expressed genes(DEGs)were identified.WGCNA was employed to identify gene modules associated with DHL.A protein-protein interaction(PPI)network was constructed to screen for key hub genes.Drug-gene association analysis was conducted using the EpiMed platform to identify potential targeted drugs for DHL.The effects of bexarotene on DHL cell proliferation and key protein expression were evaluated using the CCK-8 assay and Western blotting(WB),and its effects on cell apoptosis was evaluated using flow cytometry.Results:WGCNA identified a turquoise module highly associated with DHL,and 10 hub genes(COL1A2,COL3A1,MMP2,COL5A2,DCN,BGN,FN1,MMP9,FBN1,and LUM)were screened from the PPI network.Drug association analysis nominated bexarotene as a potential therapeutic agent.In vitro validation demonstrated that bexarotene significantly inhibited U2932 cell viability(P<0.05),promoted cell apoptosis(P<0.001),and downregulated c-Myc and COL1A2 expression(P<0.05).Conclusion:Bexarotene may exert anti-DHL effects by suppressing the c-Myc signaling pathway and modulating extracellular matrix-related genes.Further studies are warranted to validate its in vivo efficacy and potential for combination therapy.

Objective:To investigate the effect of concentrated growth factor (CGF) on the biological performance of human dental pulp stem cells (hDPSCs) under oxidative stress status induced by hydrogen peroxide (H 2O 2). Methods:The hDPSCs were isolated by using tissue block separation method from healthy permanent teeth extracted for orthodontic reason. hDPSCs surface markers CD34, CD45, CD90 and CD105 were detected by flow cytometry. Alkaline phosphatase (ALP), alizarin red S (ARS), oil red O staining and colony formation assay were used to identify hDPSCs. After the cell counting kit-8 (CCK-8) detection, the optimal H 2O 2 concentration was used to construct the hDPSCs oxidative stress model. CGF conditioned medium was prepared by repeated freeze-thaw methods. After CCK-8 detection, the optimum CGF concentration was chosen for the subsequent experiments. The hDPSCs were divided into control group, H 2O 2 (only H 2O 2 processing), H 2O 2+CGF group (H 2O 2 processing in combination with the CGF) and CGF group (only CGF processing). Subsequent experiments were performed according to these groups. The oxidative stress model was verified by reactive oxygen species, β-galactosidase staining and Western blotting. The effects of CGF on the proliferation and migration of hDPSCs under oxidative stress status were detected by CCK-8 and cell scratch assay, respectively. ALP activity and ARS staining were used to detect the effect of CGF on the osteogenic differentiation of hDPSCs under oxidative stress status. The mRNA expression levels of odontogenesis related genes were detected by real-time fluorescence quantitative PCR (RT-qPCR), and the expression levels of odontogenesis and osteogenesis related proteins were detected by Western blotting. Results:Isolated hDPSCs showed positive expression of mesenchymal stem cells surface markers of CD90, CD105, and negative expression of hematopoietic stem cells surface markers CD34, CD45. The hDPSCs were proved to have the capacity of osteogenic, adipogenic differentiation and clone formation. The optimal concentration to construct the oxidative stress model was 200 μmol/L H 2O 2. Twenty percent CGF was the optimal concentration for subsequent experiments. Compared with the control group, the expression of aging protein p53 was significantly up-regulated from (0.82±0.12) to (1.19±0.14) in H 2O 2 group ( P<0.05), with deepened β-galactosidase staining and increased fluorescence intensity of reactive oxygen species. The proliferative capacity of cells in H 2O 2+CGF group on day 1, 3, 5 and 7 (0.23±0.01, 0.50±0.02, 1.60±0.07, 1.80±0.21) were all higher than in H 2O 2 group (0.15±0.01, 0.14±0.02, 0.50±0.03, 0.90±0.09) ( P<0.05). Cell healing capacity of cells in H 2O 2+CGF group at 12 h and 24 h [(47±7)%, (58±44)%] also increased compared with the H 2O 2 group [(36±2)%, (44±2)%] ( P<0.05), and similar results in the activity of ALP and the formation of mineralized nodules. On day 28, the mRNA expressions of dentin sialophosphoprotein (0.52±0.16) and dental matrix protein 1 (DMP-1) (0.39±0.13) in H 2O 2 group were all significantly lower than those in H 2O 2+CGF group (0.96±0.24, 0.83±0.30, respectively) and CGF group (1.12±0.18, 1.23±0.19, respectively) ( P<0.05). On day 28, the expressions of odontogenesis related protein DMP-1 (0.27±0.04) and osteogenesis related protein Runt-related transcription factor-2 (0.42±0.15) in H 2O 2 group were all significantly lower than those in H 2O 2+CGF group (0.66±0.18, 0.68±0.04) and CGF group (1.15±0.13, 1.06±0.19, respectively) ( P<0.05). Conclusions:H 2O 2 can induce oxidative stress in hDPSCs, while CGF can promote proliferation, migration, odontogenic and osteogenic differentiation of hDPSCs under oxidative stress status.

Single-cell and spatial multi-omics technologies enable the simultaneous analysis of thousands of cells in various states, revealing their transcriptional profiles, chromatin accessibility, and spatial positioning. Recent advances in single-cell multi-omics have led to significant discoveries regarding the definition, function, evolution, and interaction of various cell subtypes during tooth development. We summarize key findings from recent single-cell multi-omics studies on tooth development, highlighting their contributions to the field and discussing future perspectives in dental developmental genomics research.

Objective To analyze the current status of outcome measures in randomized controlled trials(RCTs)of acupuncture treatment for vascular dementia(VD)and promote the development of a standardized set of outcome measures.Methods Chinese and English literature databases were searched,including the Chinese Medical Periodical Full-Text Database,the Chinese Biology Medicine disc,China National Knowledge Infrastructure,Wanfang Data,VIP Database,PubMed,Embase,the Cochrane Library,MEDLINE,Web of Science,Chinese Clinical Trials Registry,and the International Traditional Medicine Clinical Trial Registry.Two researchers independently screened RCT literature on acupuncture treatment for VD between January 1,2015 and January 1,2025,risk of bias was assessed using the Cochrane Risk of Bias 2 tool.Extract basic study information,intervention measures,diagnostic criteria for both Chinese and Western medicine,TCM syndromes,and outcome measures.Summarize the indicator domains of RCT studies on acupuncture treatment for VD,and analyze the basic information and outcome measures of the included studies.Results A preliminary search identified 2,898 articles,of which 93 RCTs were ultimately included.These studies involved 84 outcome measures,covering six indicator domains:symptoms/signs(23.81%),traditional Chinese medicine(TCM)syndromes(3.57%),medical checkups(60.71%),quality of life(5.95%),safety assessment(4.76%),and prognosis follow-up(1.19%).A total of 91(97.85%)RCTs reported treatment duration,ranging from 2 to 24 weeks;72(77.42%)RCTs used clinical efficacy as the outcome indicator;11 studies(11.83%)reported safety assessments and adverse events.Conclusion Currently,the RCT study design for acupuncture treatment of VD lacks unified standards and has numerous methodological issues.These include insufficient description of sample size estimation processes,strong reliance on subjective rating scales,ambiguous definitions of primary and secondary outcome measures,incomplete integration of Chinese and Western medical indicators,and insufficient reflection of individualized syndrome differentiation and treatment characteristics.In addition,safety assessments and follow-up mechanisms remain relatively weak.Future research should focus on the essential nature of VD,establish a core set of indicators aligned with the clinical characteristics of traditional Chinese medicine,promote the scientific and standardized development of acupuncture research for VD,and provide more compelling evidence-based support for clinical practice.

Abnormal accumulation of collagen fibrils is a hallmark feature of oral submucous fibrosis (OSF). However, the precise characteristics and underlying mechanisms remain unclear, impeding the advancement of potential therapeutic approaches. Here, we observed that collagen I, the main component of the extracellular matrix, first accumulated in the lamina propria and subsequently in the submucosa of OSF specimens as the disease progressed. Using RNA-seq and Immunofluorescence in OSF specimens, we screened the cartilage oligomeric matrix protein (COMP) responsible for the abnormal collagen accumulation. Genetic COMP deficiency reduced arecoline-stimulated collagen I deposition significantly in vivo. In comparison, both COMP and collagen I were upregulated under arecoline stimulation in wild-type mice. Human oral buccal mucosal fibroblasts (hBMFs) also exhibited increased secretion of COMP and collagen I after stimulation in vitro. COMP knockdown in hBMFs downregulates arecoline-stimulated collagen I secretion. We further demonstrated that hBMFs present heterogeneous responses to arecoline stimulation, of which COMP-positive fibroblasts secrete more collagen I. Since COMP is a molecular bridge with Fibril-associated collagens with Interrupted Triple helices (FACIT) in the collagen network, we further screened and identified collagen XIV, a FACIT member, co-localizing with both COMP and collagen I. Collagen XIV expression increased under arecoline stimulation in wild-type mice, whereas it was hardly expressed in the Comp^-/- mice, even with under stimulation. In summary, we found that COMP may mediates abnormal collagen I deposition by functions with collagen XIV during the progression of OSF, suggesting its potential to be targeted in treating OSF.
Oral Submucous Fibrosis/pathology* ; Cartilage Oligomeric Matrix Protein/genetics* ; Animals ; Mice ; Humans ; Fibroblasts/metabolism* ; Collagen Type I/metabolism* ; Arecoline/pharmacology* ; Mouth Mucosa/metabolism* ; Cells, Cultured ; Fluorescent Antibody Technique

Objective:To explore the specific mechanism of lactic acid(LA)on regulating decidual macrophage differentia-tion in pre-eclampsia(PE)through non-receptor tyrosine kinase sarcoma virus protein/lactate dehydrogenase A(SRC/LDHA)signaling pathway.Methods:Decidual tissues were collected from 16 women with preeclampsia(PE)and 16 women with normal pregnancies during the same period.Immunofluorescence was used to assess macrophage subtypes and LA secretion in decidual tissues.Biochemi-cal assays and Western blot were performed to measure levels of LA,LDHA and MCT-4 in tissues.Biochemical assays were conducted to measure LA level in supernatants of normal trophoblast cells under different cell densities and oxygen conditions.Flow cytometry was used to assess polarization of macrophages in normal decidual tissues under different conditions of LA,oxygen and culture media.qRT-PCR,Western blot,ROS,and NAD+/NADH measurements were performed to evaluate mRNA and protein levels related to gly-colysis and mitochondrial oxidative phosphorylation,as well as proteins associated with SRC/LDHA signaling pathway,ROS level,and NAD+/NADH.Thirty pregnant rats were divided into normal group,model group,low-,medium-,high-dose AZD3965 groups,with 6 rats in each group.PE model was established,and the systolic blood pressure and urinary protein levels of pregnant rats were measured at various time points.On gestational day 20,the placental and fetal development in each group were compared.Immunofluo-rescence was used to assess trophoblast cell invasion and uterine spiral artery remodeling in decidual tissues.Immunohistochemistry and flow cytometry were performed to evaluate fetal vascular development and macrophage polarization.Results:Compared with nor-mal pregnant women,patients with PE exhibited an increased proportion of M1 macrophages,CK7+LDHA+,CK7+MCT-4+cells,LA,LDHA and MCT-4(P<0.05),while proportion of M2 macrophages was decreased(P<0.05).Compared with normoxic conditions,LA level in supernatant of trophoblast cells was significantly increased under hypoxic conditions(P<0.05).Under normoxic condition,compared with control groups,conditioned culture group showed a decreased M1/M2 macrophage(P<0.05).LA group exhibited in-creased expressions of NDUFA1,NDUFA2,UQCRC1,UQCRC2,COX4I1,COX4I2,ATP5F1A,ATP5F1B mRNA,and SDHB,COX4I1,LDHB,p-SRC(Y416),p-LDHA(Y10),VEGF and ARG-1 protein(P<0.05).NAD+/NADH was decreased(P<0.05).PX478 group showed increased expression of p-SRC(P<0.05).LA+PX478 group exhibited increased expressions of p-SRC(Y416)and p-LDHA(Y10)(P<0.05),while VEGF expression was decreased(P<0.05).Under hypoxic conditions,compared with control groups,the conditioned culture group,LA group and LA+PX478 group exhibited an increased M1/M2 macrophage(P<0.05).LA group showed upregulated expressions of LDHA mRNA,HIF-1α,LDHA,p-SRC(Y416),p-LDHA(Y10)and iNOS(P<0.05),while mRNA expression of LDHB and protein expressions of NDUFA1 and UQCRC2 were downregulated(P<0.05).ROS level and NAD+/NADH were decreased(P<0.05).PX478 group demonstrated reduced HIF-1α expression(P<0.05)and increased p-SRC(Y416)expression(P<0.05).LA+PX478 group showed decreased HIF-1α expression(P<0.05)and increased p-LDHA(Y10)ex-pression(P<0.05).Compared with LA group,LA+PX478 group exhibited elevated ROS level(P<0.05)and a reduced NAD+/NADH(P<0.05).Compared with normal group,pregnant rats in model group exhibited increased blood pressure,urine protein levels and an elevated M1/M2 in decidual tissue(P<0.05).The fetal mice exhibited abnormal development,with inadequate trophoblast cell infiltra-tion,impaired uterine spiral artery remodeling,a decreased number of fetal blood vessels in placental labyrinthine layer,and nar-rowed lumens.Compared with model group,the low,medium,and high-dose AZD3965 groups showed decreased blood pressure,urine protein levels and M1/M2(P<0.05).Additionally,there was an improvement in fetal mouse development,trophoblast cell infil-tration and uterine spiral artery remodeling.The number of fetal blood vessels in placental labyrinthine layer increased,and the lu-mens expanded.Conclusion:LA can regulate macrophage polarization in decidua of PE through SRC/LDHA pathway,and blocking LA uptake can alleviate the pathological manifestations in pregnant rats with PE.