BACKGROUND: Bile acids (BAs) facilitate the progression of gastric intestinal metaplasia (GIM). Long non-coding RNAs (lncRNAs) dysregulation was observed along with the initiation of gastric cancer. However, how lncRNAs function in GIM remains unclear. This study aimed to explore the role and mechanism of lncRNA PVT1 in GIM, and provide a potential therapeutic target for GIM treatment. METHODS: We employed RNA sequencing (RNA-seq) to screen dysregulated lncRNAs in gastric epithelial cells after BA treatment. Bioinformatics analysis was conducted to reveal the regulatory mechanism. PVT1 expression was detected in 21 paired biopsies obtained under endoscopy. Overexpressed and knockdown cell models were established to explore gene functions in GIM. Molecular interactions were validated by dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (Ch-IP). The levels of relative molecular expression were detected in GIM tissues. RESULTS: We confirmed that lncRNA PVT1 was upregulated in BA-induced GIM model. PVT1 promoted the expression of intestinal markers such as CDX2 , KLF4 , and HNF4α . Bioinformatics analysis revealed that miR-34b-5p was a putative target of PVT1 . miR-34b-5p mimics increased CDX2 , KLF4 , and HNF4α levels. Restoration of miR-34b-5p decreased the pro-metaplastic effect of PVT1 . The interactions between PVT1 , miR-34b-5p, and the downstream target HNF4α were validated. Moreover, HNF4α could transcriptionally activated PVT1 , sustaining the GIM phenotype. Finally, the activation of the PVT1 /miR-34b-5p/ HNF4α loop was detected in GIM tissues. CONCLUSIONS BAs facilitate GIM partially via a PVT1/miR-34b-5p/HNF4α positive feedback loop. PVT1 may become a novel target for blocking the continuous development of GIM and preventing the initiation of gastric cancer in patients with bile reflux.
Humans ; RNA, Long Noncoding/metabolism* ; MicroRNAs/metabolism* ; Hepatocyte Nuclear Factor 4/genetics* ; Bile Acids and Salts ; Kruppel-Like Factor 4 ; Metaplasia/metabolism*

Bone repair remains an important target in tissue engineering, making the development of bioactive scaffolds for effective bone defect repair a critical objective. In this study, β-tricalcium phosphate (β-TCP) scaffolds incorporated with processed pyritum decoction (PPD) were fabricated using three-dimensional (3D) printing-assisted freeze-casting. The produced composite scaffolds were evaluated for their mechanical strength, physicochemical properties, biocompatibility, in vitro pro-angiogenic activity, and in vivo efficacy in repairing rabbit femoral defects. They not only demonstrated excellent physicochemical properties, enhanced mechanical strength, and good biosafety but also significantly promoted the proliferation, migration, and aggregation of pro-angiogenic human umbilical vein endothelial cells (HUVECs). In vivo studies revealed that all scaffold groups facilitated osteogenesis at the bone defect site, with the β-TCP scaffolds loaded with PPD markedly enhancing the expression of neurogenic locus Notch homolog protein 1 (Notch1), vascular endothelial growth factor (VEGF), bone morphogenetic protein-2 (BMP-2), and osteopontin (OPN). Overall, the scaffolds developed in this study exhibited strong angiogenic and osteogenic capabilities both in vitro and in vivo. The incorporation of PPD notably promoted the angiogenic-osteogenic coupling, thereby accelerating bone repair, which suggests that PPD is a promising material for bone repair and that the PPD/β-TCP scaffolds hold great potential as a bone graft alternative.
Calcium Phosphates/chemistry* ; Animals ; Bone Regeneration ; Rabbits ; Tissue Scaffolds ; Printing, Three-Dimensional ; Humans ; Human Umbilical Vein Endothelial Cells ; Neovascularization, Physiologic ; Osteogenesis ; Tissue Engineering/methods* ; Biomimetic Materials ; Cell Proliferation ; Angiogenesis

Objective To investigate the potential value of exosomes derived from rat ectoderm mesenchymal stem cells(EMSCs-exo)for repairing secondary spinal cord injury.Methods EMSCs-exo were obtained using ultracentrifugation from EMSCs isolated from rat nasal mucosa,identified by transmission electron microscope,nanoparticle tracking analysis(NTA),and Western blotting,and quantified using the BCA method.Neonatal rat microglia purified by differential attachment were induced with 100 μg/L lipopolysaccharide(LPS)and treated with 37.5 or 75 mg/L EMSCs-exo.PC12 cells were exposed to 400 μmol/L H2O2 and treated with EMSCs-exo at 37.5 or 75 mg/L.The protein and mRNA expressions of Arg1 and iNOS in the treated cells were determined with Western blotting and qRT-PCR,and the concentrations of IL-6,IL-10,and IGF-1 in the supernatants were measured with ELISA.The viability and apoptosis of PC12 cells were detected using CCK-8 assay and flow cytometry.Results The isolated rat EMSCs showed high expressions of nestin,CD44,CD105,and vimentin.The obtained EMSCs-exo had a typical cup-shaped structure under transmission electron microscope with an average particle size of 142 nm and positivity for CD63,CD81,and TSG101 but not vimentin.In LPS-treated microglia,EMSCs-exo treatment at 75 mg/L significantly increased Arg1 protein level and lowered iNOS protein expression(P<0.05).EMSCs-exo treatment at 75 mg/L,as compared with the lower concentration at 37.5 mg/L,more strongly increased Arg1 mRNA expression and IGF-1 and IL-10 production and decreased iNOS mRNA expression and IL-6 production in LPS-induced microglia,and more effectively promoted cell survival and decreased apoptosis rate of H2O2-induced PC12 cells(P<0.05).Conclusion EMSCs-exo at 75 mg/L can effectively reduce the proportion of M1 microglia and alleviate neuronal apoptosis under oxidative stress to promote neuronal survival,suggesting its potential in controlling secondary spinal cord injury.

Objective To investigate the potential value of exosomes derived from rat ectoderm mesenchymal stem cells(EMSCs-exo)for repairing secondary spinal cord injury.Methods EMSCs-exo were obtained using ultracentrifugation from EMSCs isolated from rat nasal mucosa,identified by transmission electron microscope,nanoparticle tracking analysis(NTA),and Western blotting,and quantified using the BCA method.Neonatal rat microglia purified by differential attachment were induced with 100 μg/L lipopolysaccharide(LPS)and treated with 37.5 or 75 mg/L EMSCs-exo.PC12 cells were exposed to 400 μmol/L H2O2 and treated with EMSCs-exo at 37.5 or 75 mg/L.The protein and mRNA expressions of Arg1 and iNOS in the treated cells were determined with Western blotting and qRT-PCR,and the concentrations of IL-6,IL-10,and IGF-1 in the supernatants were measured with ELISA.The viability and apoptosis of PC12 cells were detected using CCK-8 assay and flow cytometry.Results The isolated rat EMSCs showed high expressions of nestin,CD44,CD105,and vimentin.The obtained EMSCs-exo had a typical cup-shaped structure under transmission electron microscope with an average particle size of 142 nm and positivity for CD63,CD81,and TSG101 but not vimentin.In LPS-treated microglia,EMSCs-exo treatment at 75 mg/L significantly increased Arg1 protein level and lowered iNOS protein expression(P<0.05).EMSCs-exo treatment at 75 mg/L,as compared with the lower concentration at 37.5 mg/L,more strongly increased Arg1 mRNA expression and IGF-1 and IL-10 production and decreased iNOS mRNA expression and IL-6 production in LPS-induced microglia,and more effectively promoted cell survival and decreased apoptosis rate of H2O2-induced PC12 cells(P<0.05).Conclusion EMSCs-exo at 75 mg/L can effectively reduce the proportion of M1 microglia and alleviate neuronal apoptosis under oxidative stress to promote neuronal survival,suggesting its potential in controlling secondary spinal cord injury.

Objective To evaluate the risk of bias and reporting quality in randomized controlled trials(RCTs)of the Chinese medicine injection for acute cerebral infarction in the last five years.Methods RCTs literature on Chinese medicine injection in the treatment of acute cerebral infarction was systematically searched in CNKI,Wanfang Data,VIP,China Biology Medicine Database(CBM),PubMed,Embase and Cochrane Library from April 20,2018 to April 20,2023.The risk of bias and reporting quality of included RCTs were evaluated using the Cochrane Risk of Bias Tool(ROB 1.0)and CONSORT-CHM Formulas 2017,respectively.Results A total of 4 301 articles were retrieved,and 408 RCTs were included according to inclusion and exclusion criteria.The ROB evaluation results showed that the the majority of studies were rated as having an unclear risk of bias due to the lack of reporting on allocation concealment,blind method,trial registration information,and funding sources.The evaluation results of CONSORT-CHM Formulas 2017 showed that the number of reported papers of 17 items was greater than or equal to 50%,and the number of reported papers of 25 items was less than 10%,and most of the RCTs did not show the characteristics of TCM syndrome differentiation and treatment.Conclusion The quality of Chinese medicine injection in the treatment of acute cerebral infarction RCTs is generally low.It is recommended that researchers refer to the methodology design of RCTs and international reporting standards,improve the trial design,standardize the trial report,and highlight the characteristics of TCM syndrome differentiation and treatment.

Objective:To explore therapeutic efficacy and mechanism of puerarin(PUE)in treating of ulcerative colitis(UC).Methods:Network pharmacology and molecular docking technique were used to screen and analyze targets of PUE in regulating UC.C57BL/6 mice were given free access to 2.5%DSS aqueous solution for 7 days,and influence of PUE on changes in body weight and disease activity index(DAI)score were subsequently observed.Histopathological alterations of colon tissue were observed by HE staining,changes of goblet cell population in colon tissue were evaluated through Alcian blue staining;expressions of inflammatory factors in colon tissue were detected by qRT-PCR and ELISA.Effect of PUE on MODE-K cell viability and apoptosis were assessed by CCK-8 and flow cytometry.Results:A total of 38 common targets of PUE in modulating UC,such as AKT1,TNF,STAT3,CASP3,HIF1A and etc,mainly involving TNF,IL-17 and PI3K-Akt signaling pathway.In vivo experiments confirmed that PUE ameliorated degree of colon shortening,body weight and DAI scores and reduced inflammatory cell infiltration in mice.Besides,expressions of inflammatory factors in colon,such as TNF-α and IL-1β,were inhibited by PUE.Furthermore,in vitro experiments validated that PUE relieved DSS-induced apoptosis of epithelial cells.Conclusion:PUE alleviates occurrence and development of DSS-induced UC in mice.

Objective:To explore the protective effect and mechanism of acetate on sepsis-induced acute kidney injury (AKI) in rats.Methods:Male Sprague-Dawley (SD) rats were divided into sham operation group (Sham group), sepsis group caused by cecal ligation and puncture (CLP group), and acetate pretreatment group [NaA group, gavage sodium acetate (NaA) 300 mg/kg twice a day for 7 consecutive days before CLP] using a random number table method, with 7 rats in each group. The blood was taken from the main abdominal artery 24 hours after modeling, and renal tissue was collected from the rats. Enzyme linked immunosorbent assay (ELISA) was used to detect the serum levels of interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), and kidney injury molecule-1 (KIM-1). The concentration of serum acetate was determined by high performance liquid chromatography. The level of malondialdehyde (MDA) in renal tissue was detected by thiobarbituric acid method. Myeloperoxidase (MPO) in renal tissue was detected by colorimetric method. Hematoxylin-eosin (HE) staining was used to observe histopathological changes and assess renal tubule injury score. Western blotting was used to detect the protein expressions of G-protein coupled receptor 43 (GPR43) and adenosine monophosphate-activated protein kinase/silence infor-mation regulator 1/peroxlsome proliferator-activated receptor-γ coactlvator-1α (AMPK/SIRT1/PGC-1α) pathway. The positive expressions of GPR43, phosphorylation-AMPK (p-AMPK), SIRT1, PGC-1α were detected by immunohistochemistry.Results:Compared with Sham group, the serum levels of IL-6, TNF-α and KIM-1 were significantly increased in CLP group, the contents of MDA and MPO in renal tissue were increased, and the content of acetate was significantly decreased. HE staining results showed that most of the tubular epithelial cells were denaturated with local necrosis, a large number of brush border injuries and shedding, tubular structure destruction and fragmentation, and more inflammatory cells infiltrated the renal interstitium, the renal tubular injury score significantly increased. The expressions of GPR43, p-AMPK/AMPK, SIRT1, and PGC-1α in renal tissue were significantly reduced, indicating renal injury and increased levels of oxidative stress and inflammation in septic rats. Compared with the CLP group, the serum levels of IL-6, TNF-α and KIM-1 in the NaA group were decreased [IL-6 (ng/L): 126.20±6.23 vs. 161.00±17.37, TNF-α (ng/L): 85.59±7.70 vs. 123.50±17.78, KIM-1 (μg/L): 2.92±0.38 vs. 4.73±0.36, all P < 0.05]. The contents of MDA and MPO in renal tissue were significantly decreased [MDA (μmol/g): 6.56±0.18 vs. 8.53±0.34, MPO (U/g): 2.99±0.20 vs. 3.72±0.29, both P < 0.05]. HE staining showed that kidney injury had been alleviated, with a decrease in renal tubular injury score [1 (1, 2) vs. 3 (2, 3), P < 0.05]. Western blotting showed that the expressions of GPR43 and AMPK/SIRT1/PGC-1α pathway related proteins were significantly increased in renal tissue (GPR43/β-actin: 0.62±0.09 vs. 0.41±0.09, p-AMPK/AMPK: 0.58±0.07 vs. 0.44±0.06, SIRT1/β-actin: 0.85±0.06 vs. 0.73±0.03, PGC-1α/β-actin: 0.79±0.07 vs. 0.62±0.05, all P < 0.05). Immunohistochemistry showed that the positive expressions of GPR43, p-AMPK, SIRT1 and PGC-1α were significantly increased in renal tissue [GPR43 positive area: (33.66±2.62)% vs. (16.21±1.66)%, p-AMPK positive area: (16.64±2.11)% vs. (5.04±1.28)%, SIRT1 positive area: (14.61±2.86)% vs. (7.34±1.00)%, PGC-1α positive area: (15.30±2.39)% vs. (4.84±1.67)%, all P < 0.05], the serum acetate concentration significantly increased (μg/L: 32?479±14?683 vs. 12?935±3?197, P < 0.05). Conclusion:Acetate can ameliorate sepsis-induced AKI, the mechanism may be related to the activation of AMPK/SIRT1/PGC-1α pathway by GPR43.

Objective To investigate the regulatory effects of plasma-derived exosomal microRNA-1306-5p(miR-1306-5p)on inflammation,apoptosis and oxidative stress in intestinal mucosal epithelial cells in sepsis,and to explore the potential mechanisms.Methods Sepsis plasma-derived exosomes and healthy plasma exo-somes were separated and divided into healthy plasma exosomes group and sepsis plasma-derived exosomes group.The exosomes were observed by electron microscopy,the physical parameters of the two groups of exo-somes were analyzed,and the expression of miR-1306-5p in the exosomes was detected.Intestinal mucosal epi-thelial cells were divided into control group,negative control group of miR-1306-5p mimic(mimic-NC group),miR-1306-5p mimic group(mimic group),mimic combined with overexpression of PLK1 empty vector group(mimic-PLK1-EV group),and mimic combined with overexpression of Polo-like kinase 1(PLK1)group(mimic+PLK1-OE group).Real-time fluorescent quantitative PCR was used to detect the mRNA expressions of miR-1306-5p and PLK1 in each group,and protein imprinting was used to detect the expressions of miR-1306-5p target genes PLK1,caspase 3,B lymphoblastoma-2(Bcl-2)and Bcl2-associated X protein(Bax).Dual luciferase reporter gene assay was used to detect the binding effect of miR-1306-5p and PLK1,and flow cytom-etry was used to detect apoptosis.The expressions of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6 and IL-8 in the supernatant of cultured cells were detected by enzyme-linked immunosorbent assay.The expressions of reactive oxygen species(ROS),malondialdehyde(MDA),glutathione(GSH)and superoxide dismutase(SOD)were detected by kit method.Results The plasma derived exosomes of sepsis and healthy plasma were ellipsoid in shape,and there was no significant difference in physical parameters(P＞0.05).Compared with the healthy plasma exosomes group,the expression of miR-1306-5p was up-regulated in the plasma derived exosomes of sepsis group(P＜0.05).PLK1 was identified as the target gene of miR-1306-5p by double luciferase reporter method.Compared with mimic-NC group,the expressions of miR-1306-5p,TNF-α,IL-6,IL-8,IL-1β,caspase3,Bax,ROS and MDA were up-regulated,the apoptosis rate was increased,and the expressions of PKL1,Bcl-2,GSH and SOD were down-regulated in mimic group(all P＜0.05).Compared with mimic+PLK1-EV group,the expressions of TNF-α,IL-6,IL-8,IL-1β,caspase3,Bax,ROS,and MDA were significantly down-regulated,the apoptosis rate was decreased,and the expressions of PKL1,Bcl-2,GSH and SOD were up-regulated in mimic+PLK1-OE group(all P＜0.05).Conclusion Plasma derived exosome miR-1306-5p in sepsis promotes inflammation,apoptosis and oxidative stress damage of intestinal mucosal epi-thelial cells by targeting PKL1 inhibition.

OBJECTIVE To provide a detailed report and interpretation of the method and results for determining the weights of the technical indicators from the “multi-dimensional and multi-criteria comprehensive evaluation index system （first edition）” stated in Guideline for Multi-dimensional and Multi-criteria Comprehensive Evaluation of Chinese Patent Medicine. METHODS Normalization calculations were performed on the comprehensive weight values calculated by the analytic hierarchy process and expert weighting method to obtain the objective weights of the indicators. RESULTS The weight results of the six primary dimensions in the current comprehensive evaluation indicator system of Chinese patent medicine showed effectiveness dimension＞ safety dimension＞standard dimension＞application dimension＞scientific dimension＞economic dimension， with weight values of 0.281 0， 0.268 5， 0.195 8， 0.107 3， 0.096 1 and 0.051 3 respectively， consistent with the results of most researches currently. CONCLUSIONS The process of weight determination in this indicator system is scientifically reasonable， with clear methods and clear interpretations， and is worthy of further optimization and widespread application.

The retrieval and evaluation of evidence is the basis for the development of clinical practice guidelines for Chinese patent medicine. As traditional Chinese medicine has a different development trajectory and utilization characteristics from modern medicine， there is certain differences in terms of evidence composition， retrieval and integration.This paper discussed multi-source body of evidence on Chinese patent medicine based on modern evidence-based medicine and ancient medical literature， and summarized the retrieval strategy as well as the possible problems and solving methods. For different types of evidence on Chinese patent medicine， the corresponding evaluation tools have been recommended， and the order to integrate the evidence based on the quality of the evidence from high to low is suggested. Finally， a multi-source based evidence retrieval-evaluation-integration scheme for Chinese patent medicine has been formed， which will provide a methodological reference for practitioners in the development of clinical practice guidelines for Chinese patent medicine.