OBJECTIVES: This study aims to investigate the optimal dose ratio and mechanisms of the primary active components in Yinchenhao decoction (geniposide, chlorogenic acid, and rhubarb polysaccharides) for ameliorating metabolic-associated steatotic liver disease (MASLD). METHODS: C57BL/6 mice were randomly divided into the normal control group, model control group, uniform design groups 1-6, and Yinchenhao Decoction group; except for the normal control group, mice in all other groups were fed a Western diet to establish a MASLD model, and after 8 weeks of modeling, mice in the uniform design groups 1-6 and Yinchenhao Decoction group were given the corresponding drugs by gavage. At 12 weeks, all mice were sacrificed: their body weight and liver weight were measured, hematoxylin-eosin staining was used to observe the histopathological changes of liver tissue, the plasma levels of alanine transaminase (ALT) and aspartate transaminase (AST) were detected, and the levels of total cholesterol (TC) and triglycerides (TG) in plasma and liver were measured. Based on these results, the optimal uniform design group was identified; subsequently, with plasma AST, plasma TG, and liver TC levels as screening indicators, the optimal dose ratio was obtained via a regression equation, which was further verified from two dimensions, namely functional indicators and tissue morphology. Meanwhile, glucose tolerance test and insulin tolerance test were conducted to evaluate glucose metabolic homeostasis and insulin sensitivity in mice, periodic acid-Schiff staining was used to observe glycogen accumulation, quantitative reverse transcription-polymerase chain reaction was employed to detect the mRNA expression of genes related to glycolipid metabolism and bile acid metabolism, Western blotting was performed to measure the protein expression of molecules involved in bile acid metabolism, and commercial kits were used to determine the plasma levels of total bilirubin (TBIL), direct bilirubin (DBIL), and total bile acid (TBA). RESULTS: Combinations of geniposide, chlorogenic acid, and rhubarb polysaccharide all reduced the liver-to-body weight ratio, alleviated liver injury, and decreased lipid accumulation, among which the uniform design group 6 (200 mg/kg geniposide+160 mg/kg chlorogenic acid+340 mg/kg rhubarb polysaccharide) exhibited the optimal efficacy. Meanwhile, regression analysis indicated that the dosage ratio of uniform design group 6 was the optimal one for MASLD intervention. Validation experiments showed that, compared with single-drug intervention, the optimal dosage ratio resulted in significantly lower body weight, as well as lower plasma levels of ALT, AST and TC in mice (all P<0.05), along with a more pronounced reduction in the area of hepatic lipid accumulation. Mechanistic investigation experiments demonstrated that intervention with the optimal dosage ratio significantly improved glucose tolerance and insulin sensitivity in mice (all P<0.05), reduced hepatic glycogen deposition, and downregulated the mRNA expression of glycolipid metabolism-related genes such as Gsk3, G6pc, Pck1, Fbp1, Fasn, Srebp-1c, Scd1, Slc27a2, and Slc27a5 (all P<0.05); it also decreased plasma levels of TBIL, DBIL, and TBA (all P<0.05), reversed the abnormal protein expression of bile salt export pump (BSEP), farnesoid X receptor (FXR), and cholesterol-7α-hydroxylase (CYP7A1) in the liver (all P<0.05), and reversed the abnormal mRNA expression of bile acid metabolism-related genes including Nr1h4, Cyp7a1, Cyp27a1, Slc10a1, and Slco1a1 (all P<0.05). CONCLUSIONS The combination of geniposide (200 mg/kg), chlorogenic acid (160 mg/kg), and rhubarb polysaccharide (340 mg/kg) exerts the optimal ameliorative effect on MASLD in mice. This superior efficacy is presumably achieved by synergistically regulating the key nodes of glycolipid metabolism and bile acid metabolism, ultimately optimizing the therapeutic outcome.

OBJECTIVE: To initially investigate the function of neuronal pentraxin 1 (NPTX1) gene on osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). METHODS: hBMSCs were induced to undergo osteogenic differentiation, and then RNA was collected at different time points, namely 0, 3, 7, 10 and 14 d. The mRNA expression levels of key genes related with osteogenic differentiation, including runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), and NPTX1, were detected on the basis of quantitative real-time polymerase chain reaction (qPCR) technology. In order to establish a stable NPTX1-knockdown hBMSCs cell line, NPTX1 shRNA lentivirus was constructed and used to infect hBMSCs. ALP staining, alizarin red (AR) staining, and qPCR were employed to assess the impact of NPTX1-knockdown on the osteogenic differentiation ability of hBMSCs. RESULTS: The results showed that during the osteogenic differentiation of hBMSCs in vitro, the mRNA expression levels of osteogenic genes RUNX2, ALP and OCN significantly increased compared with 0 d, while NPTX1 expression decreased markedly (P < 0.01) as the osteogenic induction period exten-ded. At 72 h post-infection with lentivirus, the result of qPCR indicated that the knockdown efficiency of NPTX1 was over 60%. After knocking down NPTX1 in hBMSCs, RNA was extracted from both the NPTX1-knockdown group (sh NPTX1 group) and the control group (shNC group) cultured in regular proliferation medium. The results of qPCR showed that the expression levels of osteogenic-related genes RUNX2 and osterix (OSX) were significantly higher in the sh NPTX1 group compared with the shNC group (P < 0.01). ALP staining revealed a significantly deeper coloration in the sh NPTX1 group than in the shNC group at the end of 7 d of osteogenic induction. AR staining demonstrated a marked increase in mineralized nodules in the sh NPTX1 group compared with the shNC group at the end of 14 d of osteogenic induction. CONCLUSION NPTX1 exerts a modulatory role in the osteogenic differentiation of hBMSCs, and its knockdown has been found to enhance the osteogenic differentiation of hBMSCs. This finding implies that NPTX1 could potentially serve as a therapeutic target for the treatment of osteogenic abnormalities, including osteoporosis.
Humans ; Mesenchymal Stem Cells/cytology* ; Osteogenesis/genetics* ; Cell Differentiation/genetics* ; Nerve Tissue Proteins/genetics* ; Cells, Cultured ; C-Reactive Protein/genetics* ; RNA, Small Interfering/genetics* ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Bone Marrow Cells/cytology* ; Gene Knockdown Techniques ; Osteocalcin/metabolism* ; Alkaline Phosphatase/metabolism* ; RNA, Messenger/metabolism*

Human NAD(P)H: quinone oxidoreductase 1 (NQO1) is a flavoenzyme expressed at high levels in multiple solid tumors, making it an attractive target for anticancer drugs. Bioactivatable drugs targeting NQO1, such as β-lapachone (β-lap), are currently in clinical trials for the treatment of cancer. β-Lap selectively kills NQO1-positive (NQO1⁺) cancer cells by inducing reactive oxygen species (ROS) via catalytic activation of NQO1. In this study, we demonstrated that cryptotanshinone (CTS), a naturally occurring compound, induces NQO1-dependent necrosis without affecting NQO1 activity. CTS selectively kills NQO1⁺ cancer cells by inducing NQO1-dependent necrosis. Interestingly, CTS directly binds to NQO1 but does not activate its catalytic activity. In addition, CTS enables activation of JNK1/2 and PARP, accumulation of iron and Ca²⁺, and depletion of ATP and NAD⁺. Furthermore, CTS selectively suppressed tumor growth in the NQO1⁺ xenograft models, which was reversed by NQO1 inhibitor and NQO1 shRNA. In conclusion, CTS induces NQO1-dependent necrosis via the JNK1/2/iron/PARP/NAD⁺/Ca²⁺ signaling pathway. This study demonstrates the non-enzymatic function of NQO1 in inducing cell death and provides new avenues for the design and development of NQO1-targeted anticancer drugs.

Objective To assess the value of multimodal ultrasonography for diagnosing thyroid nodules—atypia of undetermined significance (AUS) of thyroid imaging reporting and data system (TI-RADS) categories 3 to 5. Methods A total of 90 AUS thyroid nodules in TI-RADS 3-5 categories from 88 patients underwent conventional ultrasonography, ultrasound elastography, superb microvascular imaging, and multimodal ultrasonography at the same time. With fine needle aspiration biopsy results as the gold standard, the methods were compared in terms of the sensitivity, specificity, accuracy, false positive rate (FPR), false negative rate (FNR), and area under the receiver operating characteristic curve (AUC) for diagnosing thyroid nodules. Results There were no significant differences between patients with benign and those with malignant thyroid nodules in terms of sex, age, and nodule locations (all P > 0.05), but the proportion of thyroid nodules ≤ 1 cm in diameter was significantly higher for malignant thyroid nodules than for benign thyroid nodules (χ²=9.610, P=0.002). Compared with benign nodules, malignant nodules were significantly more frequent to have low-level echoes or very low-level echoes, a blurred margin, a vertical diameter/horizontal diameter ratio of > 1, and microcalcifications or no calcifications (all P < 0.05). An ultrasound elastography score of ≥ 3 and type III vascularity on superb microvascular imaging indicated a higher possibility of malignant thyroid nodules (both P < 0.001). The multivariable logistic regression analysis showed that the size, echogenicity, margin, and vertical diameter/horizontal diameter ratio, and superb microvascular imaging type of thyroid nodules were not significant markers for benign or malignant thyroid nodules (all P > 0.05), while microcalcifications/no calcifications and an ultrasound elastography score of ≥ 3 were independent risk factors for malignant AUS nodules (both P < 0.05). The diagnostic sensitivity, specificity, accuracy, FPR, and FNR of conventional ultrasonography for AUS nodules were 91.30%, 71.40%, 62.70%, 28.60%, and 8.70%, respectively; the values for ultrasound elastography were 85.50%, 66.70%, 52.20%, 33.30%, and 14.50%, respectively; the values for superb microvascular imaging were 66.70%, 76.20%, 42.90%, 23.80%, and 33.30%, respectively; and the values for multimodal ultrasonography were 75.20%, 92.50%, 67.70%, 24.80%, and 7.50%, respectively. For distinguishing between benign and malignant AUS nodules, the AUC values of conventional ultrasonography, ultrasound elastography, superb microvascular imaging, and multimodal ultrasonography were 0.866, 0.745, 0.774, and 0.918, respectively. Conclusion Multimodal ultrasonography shows better diagnostic efficacy for AUS nodules of TI-RADS 3-5 compared with conventional ultrasonography, ultrasound elastography, and superb microvascular imaging, which can facilitate the malignancy risk stratification and management of AUS thyroid nodules.

Objective To investigate the effect of vitamin D(VD)on intestinal flora in spontaneously diabetic rats.Methods Zucker diabetic fatty rats(ZDF rats)were randomised to control(Con)group,VD control(VD)group,model(T2DM)group,and VD intervention(VD+T2DM)group.Fasting blood glucose profiles and oral glucose tolerance levels were determined in rats of each group.16S rDNA sequencing was used to assess changes in rat intestinal flora.OTU analysis(Venn diagram),α diversity analysis(chao1,observed species,PD whole tree,and shannon and simpson),βdiversity analysis(principal coordinate analysis(PCoA)),flora structure,and colony species variability analysis(linear discriminant analysis and influence factor(LEfSe)analysis)were also performed.Results VD intervention significantly improved fasting blood glucose levels and insulin resistance in T2DM rats(P＜0.05).α diversity showed no significant differences in chao1,observed species,PD whole tree,and shannon and simpson indices between T2DM and VD+T2DM groups(P＞0.05).β diversity analysis showed that the VD+T2DM group had more species similarity to the Con group than the T2DM group.The dominant bacteria of rat intestinal flora in each group were significantly different.In comparison to the T2DM group,the VD+T2DM group showed a decrease in abundance of Bacteroidetes and increases in abundances of Firmicutes and Clostridium XIVa.Conclusions VD improves fasting glucose elevation and insulin resistance in T2DM rats.VD improves the structure of intestinal flora,decreases Bacteroidetes,and elevates Firmicutes and Clostridium XIVa abundances in T2DM rats.

Objective To clarify and analyze the concept and connotation of the reflective practice in nurses.Methods CNKI,Wanfang,CBM,PubMed,Embase,Scopus,CIHNAL database were searched in relation to reflective practice in nurses.The retrieval period was from the inception to Dec.2022.Walker and Avant's concept analysis method was used to analyze the connotation.Results A total of 43 articles were retrieved.6 attributes of reflective practice in nurses were identified,including active engagement,critical thinking,openness and acceptance,making connections with previous knowledge or experiences,extemal support,dynamic and continuity.Antecedents included internal and extemal drivers.The reflective practice in nurses ultimately had a vital impact on themselves,patients and organizations.In recent years Reflective Practice Questionnaire has been one of the most commonly used data collection tools to measure the reflective practice in nurses.Conclusion The concept attributes of reflective practice in nurses were identified by concept analysis method,which provides references for nursing educators to cultivate reflective practice ability of nurses.In future research,researchers can combine the connotations of reflective practice for nurses to develop specific assessment tools for testing the level of reflective practice of nurses and construct the local reflective practice intervention scheme for nurses,so as to promote the development of reflective practice in nursing field in China.

Objective: To describe and analyze the current research status of compliance in orthokeratology among children and adolescents in and beyond China, so as to guide future research in this field. Methods: By combining subject words with free words, the paper systematically searched the research related to the compliance of children and adolescents wearing orthokeratology lenses in PubMed, Web of Science, Embase, Scopus, Cochrane Library, China National Knowledge Infrastructure, Wanfang, VIP and China Biomedical Literature Database. The retrieval time limit was until September 1, 2023.Two researchers independently screened and extracted data. Results: A total of 35 articles were included, and four themes were identified:the current level of compliance, evaluation indicators, influencing factors, and intervention programs. The evaluation indicators of orthokeratology lenswearing compliance were diverse, and the influencing factors involved general demographic characteristics, personal internal factors, external factors, and specific clinical situations related to orthokeratology. Conclusions Multiple factors are associated with compliance in orthokeratology. Eyecare practitioners should choose suitable evaluation tools based on specific conditions and implement diverse and practical intervention strategies guided by theory to enhance the safety and effectiveness of orthokeratology lenswearing among children and adolescents.

Cardiovascular diseases are now the leading cause of serious harms to human health,have drawn widespread attention both domestically and internationally. But the current research on mechanism of cardiovascular diseases is not keeping up with the current status of their treatment. The microbiota in the gut,the largest microecological system in the human body and its interaction with the human host have been implicated in a variety of diseases. The relationship between gut microbiota and cardiovascular diseases has been increasingly understood in recent years. The changes of gut microbiota and its metabolites are the major contributing factor for the occurrence and development of cardiovascular diseases,therefore,correction of gut microbiota dysbiosis may provide a novel therapeutic alternative for cardiovascular diseases. This article reviews the role of gut microbiota and its metabolites in cardiovascular diseases,aiming to provide reference for future related studies.

Objective:To detect the level of the circulating cell-free mitochondrial DNA (ccf-mtDNA) in plasma of children with primary carnitine deficiency (PCD)-associated cardiomyopathy and evaluate its clinical significance.Methods:In this prospective case-control study, peripheral blood samples were collected from 7 PCD patients with cardiomyopathy (PCD group), 16 dilated cardiomyopathy (DCM) patients (DCM group), and 50 healthy children (healthy control group) in the Pediatric Cardiovascular Department Ward of First Hospital of Jilin University from July 2017 to June 2022.The ccf-mtDNA levels were measured and compared between groups by the real-time fluorescence quantitative polymerase chain reaction.The correlations between plasma ccf-mtDNA level and blood free carnitine level and cardiac function in the PCD group were analyzed.The changes in the ccf-mtDNA level were monitored after L-carnitine treatment in the PCD group.The Kruskal-Wallis test was used for comparison among the three groups.The Mann-Whitney test was used for comparison between the PCD group and the control group.Changes before and after treatment in the PCD group were analyzed using the paired Wilcoxon rank sum test.The correlation between variables was evaluated by Logistic regression.Results:The plasma ccf-mtDNA levels in the PCD and DCM groups were 3.69×10 6 (1.09×10 6-7.26×10 6) copies/L and 0.99×10 6 (0.25×10 6-4.10×10 6) copies/L, respectively, which were significantly higher than that in the healthy control group[0.09×10 6 (0.01×10 6-0.35×10 6) copies/L]( H=33.34, 24.69; all P<0.01). Besides, the plasma ccf-mtDNA level in the PCD group was higher than that in the DCM group ( H=6.31, P<0.05). In the PCD group, the plasma ccf-mtDNA level was negatively correlated with the blood free carnitine level and left ventricular ejection fraction ( r=-0.85, -0.82, all P<0.05) and positively correlated with the modified Ross score and the N-terminal pro B type natriuretic peptide level ( r=0.81, 0.83, all P<0.05) before L-carnitine treatment.After treatment, the plasma ccf-mtDNA level decreased, and the blood free carnitine level and cardiac function recovered in the PCD group.The plasma ccf-mtDNA level declined sharply from the 3 rd month[0.96×10 6(0.50×10 6-2.27×10 6) copies/L] after treatment ( Z=2.24, P<0.05) and got to 0.27×10 6 (0.18×10 6-0.76×10 6) copies/L, 0.29×10 6(0.19×10 6-0.78×10 6) copies/L, and 0.16×10 6(0.10×10 6-1.06×10 6) copies/L at the 6 th, 9 th, and 12 th months after treatment, respectively, with no statistically significant difference compared to the healthy control group[0.09×10 6(0.01×10 6-0.35×10 6) copies/L] ( Z=1.23, 1.09, 2.12; all P>0.05). Conclusions:Plasma ccf-mtDNA may act as one pathogenic factor of cardiomyopathy in PCD, and monitoring its level is clinically important for heart condition assessment in PCD.