BACKGROUND:Studies have reported that alendronate intake significantly increases bone mineral density in patients with osteoporosis. OBJECTIVE:To analyze and compare the changes in metabolites before and after alendronate intervention in ovariectomized rats by chromatography-mass spectrometry,and to further explore the specific mechanism and target of alendronate in the treatment of osteoporosis. METHODS:A total of 36 female Sprague-Dawley rats were randomly divided into model group,alendronate sodium group and sham operation group.The osteoporosis model was established by ovariectomy in the first two groups.Four weeks after modeling,the rats in the alendronate group were intragastrically given alendronate sodium,while those in the sham operation group and model group were given equal volume of normal saline.After 12 weeks of continuous gavage,the metabolites of the lumbar spine were analyzed by chromatography-mass spectrometry,and the common differential metabolites were obtained,which were analyzed by bioinformatics such as Kyoto Gene and Genome Encyclopedia pathway. RESULTS AND CONCLUSION:Totally 17 different metabolites were obtained in the three groups.The enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes showed that alendronate sodium could regulate unsaturated fatty acid biosynthesis,linoleic acid metabolism and other pathways to protect ovariectomized rats.These results indicate that alendronate sodium may exert its anti-osteoporosis effect by interfering with unsaturated fatty acid bioanabolism and linoleic acid metabolism,so as to achieve the purpose of preventing osteoporosis

Chemical investigation of the stems of Fritillaria unibracteata P.K. Hsiao & K.C. Hsia resulted in the isolation of nine compounds, by means of silica gel column chromatography, and preparative HPLC. Based on spectroscopic and chemical evidence, these compounds were identified as: 27-hydroxychlorogenone (1), sieboldogenin (2), (3β, 25S)-spirost-5-ene-3,17,27-triol (3), laxogenin (4), tigogenone (5), cerevisterol (6), ergosterol peroxide (7), stigmaterol (8), and β-sitosterol (9). Compound 1 was a new compound, and compounds 2-9 were isolated from the stems of Fritillaria unibracteata for the first time. The inhibitory effects of compounds 1−9 on A549 cells were determined using the MTT method. The results show that compound 6 exhibits moderate inhibitory activity with an IC50 value of (14.16 ± 1.11) μmol/L.

OBJECTIVES: To analyze the molecular mechanism of Xixian Pills for treatment of rheumatoid arthritis (RA). METHODS: Forty-eight rats were randomized into 6 groups (n=8), including a normal control group, a collagen-induced arthritis (CIA) model group, 3 Xixian Pills treatment (200, 400 and 800 mg/kg) groups, and a Tripterygium glycosides tablet (TGT) treatment group. In the latter 4 groups, the rats were treated with daily gavage of Xixian Pills or TGT 2 weeks after CIA modeling for 3 consecutive weeks. The differentially expressed proteins in high-dose Xixian Pills group and the model group compared with the normal control group were screened based on the tandem mass spectrometry tag (TMT) technology, and the core targets and signaling pathways were analyzed. The immune cell infiltration and gene expression data were analyzed using ggplot2 and tidyverse packages, and the correlation coefficients between the core targets and the immune cells were calculated. RESULTS: The CIA rats showed significantly increased serum levels of TNF-α and IL-6 and lowered serum IL-10 level. Treatments with high- and medium-dose Xixian Pills and TGT all significantly reduced serum TNF‑α and IL-6 and increased IL-10 levels in CIA rats. Proteomic analysis identified 160 differential proteins between the model group and high-dose Xixian Pills group, and the core targets included CCL5, STAT1, GZMB and IL7R. The areas under the ROC curve of CCL5 and STAT1 were both greater than 0.9. Immunohistochemical and immunofluorescence staining revealed increased levels of CCL5 and STAT1 in the ankle joints of CIA rats, which were significantly decreased after treatment with Xixian Pills. CONCLUSIONS Treatment with Xixian Pills offers protection of the joints in CIA rats possibly by inhibiting joint inflammation via regulating protein expressions of CCL5 and STAT1.
Animals ; Drugs, Chinese Herbal/pharmacology* ; Rats ; Arthritis, Rheumatoid/metabolism* ; Proteomics ; Tripterygium/chemistry* ; Arthritis, Experimental/metabolism* ; Tumor Necrosis Factor-alpha/blood* ; Interleukin-10/blood* ; Interleukin-6/blood* ; Male ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVE: To investigate the therapeutic effects of Banxia Xiexin Decoction (BXD), a herbal medicine formula, on inflammation and the imbalance of the gut microbiota in a rat model of colorectal cancer (CRC) induced by azoxymethane (AOM) /dextran sulfate sodium (DSS). METHODS: A total of 75 male C57BL/6 mice were randomly divided into five groups: normal control group (NC), model group (MODEL), low-dose BXD treatment group (L-BXD), high-dose BXD treatment (H-BXD) group and MS treatment group (MS). BXD and MS were used in CRC mice at the doses of 3.915 g/kg, 15.66 g/kg, 0.6 g/kg for 3 weeks consecutively. Histopathological changes in the colon were observed using hematoxylin-eosin (HE) staining. The content of inflammatory factors in serum was detected by an enzyme-linked immunosorbent assay (ELISA), and the expression of mRNA and protein of genes related to immunity, apoptosis, inflammation, and inflammatory factors was evaluated. Changes in the intestinal flora of mouse fecal were determined based on high-throughput sequencing of the 16S rRNA microbial gene. RESULTS: Compared to the model group, the low-dose BXD and high-dose BXD groups decreased the number of colon tumors, reversed weight loss, and shortened colon length of mice. The pathological examination showed that BXD alleviated the malignancy of intestinal tumors. It also suppressed signal transducer and activator of transcription 3 (STAT3), matrix metalloproteinase-9 (MMP-9), and transforming growth factor beta 1 (TGF-β1) expression, while increasing the expression of the tight junction protein ZO-1 in colon tissues. Additionally, the levels of key pathway proteins involved in inflammation (phosphorylated-STAT3, Bcl-2, COX-2) and cell cycle regulatory molecules (c-Myc and PCNA) were reduced. According to 16S rRNA sequence analysis, BXD enhanced the relative abundance of potentially beneficial bacteria, while that of cancer-related bacteria decreased. CONCLUSION BXD plays a preventive role in developing colorectal cancer; its mechanisms are related to the inhibition of inflammation and tumor proliferation, as well as maintenance of intestinal homeostasis.

Screening carbonyl reductases with the ability to catalyze the reduction of complex carbonyl compounds is of great significance for the biosynthesis of R-tolvaptan(R-TVP). In this study, the target carbonyl reductase in the crude enzyme extract of rabbit liver was separated, purified, and identified by ammonium sulfate precipitation, gel-filtration chromatography, ion exchange chromatography, affinity chromatography, and protein mass spectrometry. With the rabbit liver genome as the template, the gene encoding the carbonyl reductase rlsr5 was amplified by PCR and the recombinant strain was successfully constructed. After RLSR5 was purified by affinity chromatography, its enzymatic properties were characterized. The results indicated that the gene sequence of rlsr5 was 972 bp, encoding a protein with a molecular weight of 40 kDa. RLSR5 was a dimeric protein, and each monomer was composed of a (α/β)₈-barrel structure. RLSR5 could asymmetrically reduce 7-chloro-1-[2-methyl-4-[(2- methylbenzoyl)amino]benzoyl]-5-oxo-2,3,4,5-tetrahydro-1H-1-benzazepine (prochiral ketone, PK) to synthesize R-TVP. The specific activity of the enzyme was 36.64 U/mg, and the optical purity of the product was 99%. This enzyme showcased the optimal performance at pH 6.0 and 30 °C. It was independent of metal ions, with the activity enhanced by Mn²⁺. This study lays a foundation for the biosynthesis of tolvaptan of optical grade.
Animals ; Rabbits ; Alcohol Oxidoreductases/biosynthesis* ; Recombinant Proteins/metabolism* ; Escherichia coli/metabolism* ; Liver/enzymology*

Objective:To explore the clinical characteristics and risk factors of acute kidney injury (AKI) in patients with diquat mixed with paraquat poisoning.Methods:We retrospectively analyzed the clinical data of 210 patients admitted to the department of emergency at our hospital with paraquat, diquat or mixed poisoning from January 20, 2016, to June 10, 2024. Based on the detection results of plasma toxicants, patients were categorized into three groups: the paraquat group (PQ group), and the diquat group (DQ group), the diquat-paraquat mixed group (mixed group). We compared clinical indices, occurrence, and severity of AKI among these groups. In the mixed group, patients were further divided into AKI and non-AKI subgroups based on the presence of AKI during hospitalization, and differences in clinical indices between these subgroups were analyzed. Univariate logistic regression and least absolute shrinkage and selection operator (Lasso) regressions were used to screen risk factors, and multivariate logistic regression was applied to establish the model. ROC curves were generated, and factors influencing AKI occurrence in the mixed group were identified.Results:A total of 88 patients were included in the PQ group, 28 in the DQ group, and 57 in the mixed group. Significant differences were observed among the three groups in terms of age, mortality rate, ingestion amount, body mass index(BMI), occurrence of AKI, the incidence of organ support therapy, SIRS score, PSS score, and APACHE Ⅱ score on admission ( P < 0.05). All three groups exhibited various degrees of AKI, with the mixed group showing a higher proportion and more severe conditions. In the mixed group, compared with the non-AKI group, the AKI group showed significantly higher values for age, number of deaths, ingestion amount, SIRS score, PSS score, APACHE Ⅱ score, plasma PQ concentration on admission, plasma DQ concentration on admission, white blood cell count (WBC), neutrophil count (NEUT), monocyte count (MONO), serum creatinine (SCr), procalcitonin (PCT), c-reactive protein (CRP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood lactic acid (Lac), and cystatin C (CysC) ( P < 0.05). Conversely, the estimated glomerular filtration rate (eGFR) and partial pressure of carbon dioxide (PCO2) were significantly lower in the AKI group ( P < 0.05). Additionally, in the mixed group, SCr levels at various times post-poisoning were significantly higher compared with the non-AKI group ( P < 0.05), showing an increasing trend and peaking at 212.19 ± 101.67 μmol/L on the fifth day after poisoning. Age, ingestion amount, SIRS score on admission, WBC, MONO, and PCO 2 were extracted by Lasso-Logistic regression. Multivariate logistic regression identified ingestion amount and SIRS score on admission as the independent risk factors for the occurrence of AKI in the mixed group. The area under the ROC curve for ingestion amount and SIRS score on admission was 0.991 (95% CI: 0.976-1.000), the sensitivity was 0.940 and the specificity was 1.000. Conclusions:The diquat mixed with paraquat poisoning is associated with a higher incidence and greater severity of AKI compared with poisoning with either PQ or DQ alone. Additionally, ingestion amount and SIRS score on admission have been identified as independent risk factors for the occurrence of AKI in patients with the mixed poisoning. The combined assessment of these two factors improves AKI prediction in patients with the mixed poisoning.

Abstract Long noncoding RNA (lncRNA) is closely related to the pathogenesis of cancer,representing a burgeoning field in tumor research in recent years. MAGI2-AS3, a tumor-associated lncRNA, exerts pivotal roles in epigenetics, transcription and post-transcriptional regulation. Studies have suggested that MAGI2-AS3 may be involved in multiple stages of tumor development, and has potential applications for tumor diagnosis, therapy and prognosis. This review summarizes the expression, regulatory mechanism and clinical application value of MAGI2-AS3 in common malignant tumrs, providing the reference for tumor prevention and treatment.

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori(Hp)infection in mice.Methods L.paracasei TK1501 was cultured for 32 h at 37℃in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×107 CFU/mL.The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption,cation exchange chromatography and HPLC,and its stability and antibacterial activity were assessed.The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection,which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL.Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments,and gastric tissues of the mice were collected for HE staining.Results L.paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid,base,and organic solvents.In the in vitro experiment,the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus,Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract.In the mouse models,treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice.Conclusion L.paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.

Objective:To investigate the genomic profiles and immune microenvironment of olfactory neuroblastoma (ONB).Methods:Nineteen ONB cases diagnosed in the Beijing Tongren Hospital from May 2018 to October 2022 were divided into low-grade and high-grade groups according to the Hyams grading system, including 7 low-grade and 12 high-grade ONB. Whole exome sequencing and multiplex immunofluorescence analyses were performed on tissue samples of these ONB.Results:A total of 929 nonsynonymous alterations were identified in 18 of the 19 ONB (18/19) cases. The most commonly altered cancer-related genes were CTNNB1 (3/19) and ZNRF3 (3/19). The most mutated oncogenic pathways were the WNT and RAS pathways. The median tumor mutation burden (TMB) was 0.45/Mb, ranging from 0 to 3.25. The median tumor neoantigen load (TNB) was 9.39 neoantigens/Mb, ranging from 0 to 38.30. The median allelic mutation tumor heterogeneity (MATH) score was 16.95, ranging from 3.05 to 117.47. Only one of the 19 cases expressed PD-L1 (composite positive score, CPS>1) in the tumor cells. The median percentage of CD8 + tumor-infiltrating lymphocyte (TIL) in the tumor region was 1.08%. No significant differences were observed between the low-and high-grade groups for mutant genes, mutant pathways, TMB, TNB, MATH, PD-L1 expression levels, or CD8 + TILs percentage( P>0.05). However, the low-grade group showed significantly more CD68 + macrophages in both the tumor and total region than the high-grade group. Notably, CD68 +CD163 - macrophages accounted for an average of 80.52% of CD68 + macrophages. Conclusions:CTNNB1 and ZNRF3 are the most commonly altered cancer-related genes. The low expression of PD-L1 and the low percentage of CD8 + TIL indicate that ONB might not be sensitive to immunotherapy. The percentage of M1-type macrophages in low-grade ONB is significantly higher than that in high-grade ONB, suggesting that M1-type macrophages may be involved in the progression of ONB from low-grade to high-grade.

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori(Hp)infection in mice.Methods L.paracasei TK1501 was cultured for 32 h at 37℃in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×107 CFU/mL.The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption,cation exchange chromatography and HPLC,and its stability and antibacterial activity were assessed.The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection,which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL.Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments,and gastric tissues of the mice were collected for HE staining.Results L.paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid,base,and organic solvents.In the in vitro experiment,the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus,Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract.In the mouse models,treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice.Conclusion L.paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.