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 analyze the clinical features and genetic basis of a child with Disorder of sex development (DSD).Methods:A child who was admitted to the Linyi People′s Hospital for primary amenorrhoea on July 29, 2019 was selected as the study subject. Clinical data of the child was collected. Chromosomal karyotyping and quantitative real-time PCR were used to detect Y chromosome microdeletions and other chromosomal aberrations. Next-generation sequencing was carried out for the child and her parents. Candidate variant was verified by Sanger sequencing and bioinformatic analysis.Results:The child, a 13-year-old girl, has featured primary amenorrhoea and onset of secondary sex characteristics of males. Ultrasound exam had detected no uterus and definite ovarian structure, but narrow band vaginal hypoecho and curved cavernoid structure. The child was found to have a 46, XY karyotype without an AZF deletion. DNA sequencing revealed that she has harbored a maternally derived c. 323delA (p.Q108Rfs*188) variant in the nuclear receptor subfamily 5 group A member 1 ( NR5A1) gene, which may result in a truncated protein. The variant was classified as pathogenic (PVS1+ PM2_Supporting+ PP4) based on the guidelines from the American College of Medical Genetics and Genomics. Conclusion:The NR5A1: c. 323delA variant probably underlay the pathogenesis of 46, XY DSD in this child. The discovery of the novel variant has enriched the mutational spectrum of NR5A1 gene and provided a basis for clinical diagnosis, treatment and prenatal diagnosis.

Due to the non-targeted release and low solubility of anti-gastric cancer agent,apatinib(Apa),a first-line drug with long-term usage in a high dosage often induces multi-drug resistance and causes serious side effects.In order to avoid these drawbacks,lipid-film-coated Prussian blue nanoparticles(PB NPs)with hyaluronan(HA)modification was used for Apa loading to improve its solubility and targeting ability.Furthermore,anti-tumor compound of gamabufotalin(CS-6)was selected as a partner of Apa with reducing dosage for combinational gastric therapy.Thus,HA-Apa-Lip@PB-CS-6 NPs were constructed to synchro-nously transport the two drugs into tumor tissue.In vitro assay indicated that HA-Apa-Lip@PB-CS-6 NPs can synergistically inhibit proliferation and invasion/metastasis of BGC-823 cells via downregulating vascular endothelial growth factor receptor(VEGFR)and matrix metalloproteinase-9(MMP-9).In vivo assay demonstrated strongest anti-tumor growth and liver metastasis of HA-Apa-Lip@PB-CS-6 NPs adminis-tration in BGC-823 cells-bearing mice compared with other groups due to the excellent penetration in tumor tissues and outstanding synergistic effects.In summary,we have successfully developed a new nanocomplexes for synchronous Apa/CS-6 delivery and synergistic gastric cancer(GC)therapy.

Objective To construct a prognostic risk model for colorectal cancer(CRC)with good prediction ability based on upda-ted ferroptosis-related genes.Methods Transcriptome levels,clinical information and updated ferroptosis-related genes of CRC pa-tients were downloaded from UCSC Xena and FerrDb V2databases.The genes related to prognosis were screened out by R language and univariate COX regression analysis,and the final prognostic risk model was established by LASSO regression combined with multivariate COX regression analysis.Then,survival analysis and the predictive power of the model were assessed by R language"survival"package,"timeROC"package and COX regression.Results There were 8differentially expressed ferroptosis-related genes including NEDD4L,TIMP1,KIF20A,GDF15,ENO3,TERT,MS4A15,HOTAIR which composed of the prognostic risk model.The model divided the CRC patients into two groups,and the difference in survival prognosis was between the two groups was statistically significant(P＜0.05).The area under the receiver operating characteristic(ROC)curves of the model in 1,3 and 5 year were 0.732,0.702 and 0.738,respective-ly.The results of GO and KEGG analysis showed that the genes between the two groups were enriched in pathways and functions such as tumor process,inflammatory response,and even immunity.Immune-related functions,immune cell infiltration,and immune check-points were significantly different between high and low risk groups(P＜0.05).Conclusion The prognostic model constructed based on 8differentially expressed ferroptosis-related genes has an independent prognostic effect on CRC,providing a new way to predict the prog-nosis of CRC.

Objective To establish a method for determining the amount of silicone oil on the surface of rubber closures,which is one of the packaging material standards in the Pharmacopoeia of the People's Republic of China.Methods The development process of the method for determining silicone oil content on the surface of rubber closures was described,taking the infrared transmission mode as example.Through method validation,the scientific and rational aspects of the method were examined.Key points of the determination method were analyzed,including the selection of quantitative peaks,selection of measurement solvent,selection of extraction solvent and conditions,selection of instrument measurement methods,selection of liquid cell windows for infrared transmission mode,selection of quantification methods,and selection of dimethyl silicone oil reference standards.Results The infrared spectroscopy mode was chosen to determine the amount of polydimethylsiloxane on the surface of rubber closures based on the peak height of the Si-CH3 bond vibration absorption at 1 260 cm-1.Cyclohexane was used as the measurement solvent.The same concentration of polydimethylsiloxane reference solution was measured using both infrared transmission mode and attenuated total reflection mode,and quantification was performed based on peak height.The results from the two measurement modes were essentially the same.The linear range of polydimethylsiloxane was about 0.3-10.0 mg·mL-1,and its correlation coefficient was 0.999 8.The average recovery of the method was 102.3%-105.6%.The LOD of polydimethylsiloxane was 0.1 mg·mL-1.The silicone oil measurement results of seven batches for rubber closures were consistent with the silicone oil amounts provided by the manufacturer.Conclusion Enterprises can choose either the infrared transmission mode or the attenuated total reflection mode to measure the amount of polydimethylsiloxane on the surface of rubber closures based on their actual conditions.

Biliary fibrosis (BF) is the result of pathological repair of bile tract injury, characterized by thickening and sclerosis of the bile duct wall and progressive stricture of the lumen, which may ultimately lead to serious adverse outcomes such as biliary obstruction, biliary cirrhosis, liver failure, and hepatobiliary malignancies. Current research describes BF as a pathological feature of certain bile tract diseases, lacking a systematic summary of its etiology, pathophysiology, molecular mechanisms, and treatment. BF is a common but easily neglected disease state in biliary system, which may promote the development and progression of hepatobiliary diseases through abnormal repair mechanism after pathological biliary tract injury. Based on the latest research progress from both domestic and international perspectives, the authors review the concept, clinical manifestation, etiology, pathogenesis, and therapeutic strategies of BF to provide a reference for clinical physicians.

Objective:To examine the distribution features of angiotensin converting enzyme ( ACE) gene I/D polymorphisms and their correlation with left ventricular hypertrophy (LVH) in children and adolescents with essential hypertension. Methods:This was a case-control study.One hundred and forty-four children and adolescents who were diagnosed with essential hypertension but received no treatment at the Department of Cardiology, Children′s Hospital, Capital Institute of Pediatrics from January 2022 to April 2023 were recruited. ACE gene polymorphisms were detected using the polymerase chain reaction-restriction fragment length polymorphism method.Patients were divided into the DD+ DI genotype group and the II genotype group according to the dominant model, and the differences in gene frequencies, genotype frequencies, clinical data, and plasma renin-angiotensin-aldosterone system (RAAS) levels were compared between the two groups.In addition, according to the echocardiographic findings, patients were divided into the LVH and non-LVH groups.The risk factors of LVH were analyzed using the multivariate Logistic regression model.The correlation of ACE gene I/D polymorphisms with LVH in children and adolescents with essential hypertension was evaluated using the Spearman analysis.The correlation between plasma RAAS levels and LVH was assessed using the partial correlation analysis. Results:The frequencies of DD, DI and II genotypes in 144 patients were 11.1%, 50.7% and 38.2%, respectively.The frequency of the D and I allele was 36.5%(105/288) and 63.5%(183/288) respectively.Patients in the DD+ DI genotype group (89 cases) were older than those in the II genotype group (55 cases) ( Z=-2.308, P=0.021), and there was no statistically significant difference between the two groups in gender and height (all P>0.05).The incidence of LVH in the DD+ DI genotype group was higher than that in the II genotype group, with a statistically significant difference ( χ2=5.230, P=0.022).Renin activity in RAAS levels was positively correlated with left ventricular mass index and relative wall thickness ( r=0.276, 0.247; P=0.002, 0.006).Multivariate Logistic regression analysis showed that both DD+ DI genotype ( OR=5.678, 95% CI: 1.623-19.872) and body mass index (BMI) ( OR=1.124, 95% CI: 1.024-1.233) were risk factors for the development of LVH in children and adolescents with essential hypertension (all P<0.05). Conclusions:ACE gene I/D polymorphisms are strongly associated with LVH.DD+ DI genotype and BMI are independent risk factors for the development of LVH in children and adolescents with essential hypertension.