OBJECTIVE To investigate the absorption characteristics and tissue distribution of silybin （Sy） nanocrystals prepared by two methods in different intestinal segments of rats. METHODS Sy nanocrystals （i.e. Sy-NS-G and Sy-NS-F） with comparable particle sizes were prepared using high-pressure homogenization and anti-solvent precipitation methods， respectively. Rats were randomly divided into three groups： Sy raw drug group， Sy-NS-G group， and Sy-NS-F group. Each group was further divided into three subgroups with low， medium， and high （60， 120， 180 μg/mL） mass concentrations （calculated based on Sy）， with 3 rats in each subgroup. The absorption rate constant （Ka） and apparent absorption coefficient （Papp） of Sy raw drug， Sy-NS-G and Sy-NS-F in different intestinal segments were investigated by using the in vivo one-way intestinal perfusion experiment. Additionally， the rats were divided into three groups: Sy raw drug group， Sy-NS-G group， and Sy-NS-F group， with 20 rats in each group. Rats in each group were administered a single intragastric dose of 50 mg/kg （calculated based on Sy）. They were sacrificed at 0.3， 1， 4， 10， and 24 hours post-administration respectively， to investigate the tissue distribution of Sy raw drug， Sy- NS-G， and Sy-NS-F in the heart， liver， spleen， lungs， kidneys， brain and intestines. RESULTS In duodenum and jejunum， the Ka and Papp of the nanocrystals prepared by the two methods remained unchanged with the increase of Sy concentration， and there was no significant difference （P＞0.05）； the absorption of Sy-NS-F in the duodenum was greater than that of Sy-NS-G； the absorption sites of Sy-NS-G and Sy raw drug were mainly in the ileum， while those of Sy-NS-F were mainly in the duodenum and ileum. The concentrations of Sy-NS-G and Sy-NS-F in different tissues of rats were different； Sy-NS-G peaked in most tissues at 1 h， and the distribution concentration was as follows： intestine＞spleen＞heart＞lungs＞liver≈brain＞kidneys. Sy-NS-F reached its peak at 1 h， and the distribution concentration was in the order of intestine＞spleen＞kidney＞lung＞heart≈liver＞brain. CONCLUSIONS The absorption mode of Sy nanocrystals in the duodenum and ileum is mainly passive diffusion. In the duodenum， the absorption of Sy-NS-F is greater than that of Sy-NS-G； there are significant differences in the tissue distribution of Sy-NS-G and Sy-NS-F in rats.

[This corrects the article DOI: 10.1016/j.apsb.2021.07.004.].

In recent years, potato scab caused by Streptomyces scabies is aggravating year by year, becoming an industrial problem urgently to be resolved. Screening antagonistic bacteria with good inhibitory effect and wide adaptability is the main measure to realize effective prevention and control of the disease. This study screened three strains of antagonistic bacteria DXT2-4, T2-1 and 21-14 with good inhibitory effect on S. scabies by using plate standoff test, and identified them as Bacillus altitudinis, Bacillus safensis and Bacillus pumilus, respectively, based on morphological characteristics, physiological and biochemical properties, and 16S rRNA gene sequences. DXT2-4, T2-1 and 21-14 showed the pot control efficacy of 68.83%, 48.57%, and 57.14%, respectively. The field control efficacy of the three strains was 59.48%, 34.58% and 51.75% in Hulun Buir, Inner Mongolia Autonomous Region and 55.14%, 36.05%, and 49.05% in Huizhou, Guangdong. The three strains could grow normally in the media with pH 1.0-13.0 and with 1%-11% NaCl, and they had inhibitory effects on Rhizoctonia solani, Verticillium dahliae, Alternaria solani, and Fusarium oxysporum. The indole-3-acetic acid yields of DXT2-4, T2-1, and 21-14 were 2.23, 1.11, and 1.67 mg/L, respectively. DXT2-4 and 21-14 demonstrated strong abilities to solubilize phosphorus. The optimal carbon source, nitrogen source, and inorganic salt for fermentation of strain DXT2-4 were 2% molasses+2% corn starch, 2% soybean meal, and 0.3% MgSO₄·7H₂O, respectively. These findings suggest the three strains of bacteria can efficiently inhibit the growth of S. scabies and have strong environmental adaptability. Particularly, DXT2-4 has the best effects of inhibiting the disease and promoting plant growth, showing a high development value and broad application prospects, this is of great significance for promoting sustainable potato production and ensuring the environmentally sound utilization of resources.
Streptomyces/metabolism* ; Fermentation ; Plant Diseases/prevention & control* ; Solanum tuberosum/growth & development* ; Bacillus/growth & development* ; Antibiosis

Objective To explore the effects of long non-coding RNA(lncRNA)HEM2M overexpression on liver injury in mice with non-alcoholic fatty liver disease(NAFLD).Methods Wild-type C57BL/6(WT)mice and myeloid cell-specific HEM2M knock-in(MYKI)mice were fed normal(ND)or high-fat diet(HFD)for 12 weeks.After intraperitoneal glucose tolerance and insulin tolerance tests,the mice were euthanized for detection of liver function indicators in the serum and liver tissue.HE staining and F4/80 immunohistochemical staining were used to examine liver pathologies,and the levels of IL-6,IL-1β,and TNF-α in the liver tissues were determined with ELISA.The mRNA expressions of HEM2M and the markers of M1 macrophages(TNF-α,iNOS,and IL-6)and M2 macrophages(Arg-1,YM-1,and IL-10)were detected using qRT-PCR,and the protein expressions of P-AKT,T-AKT,NLRC4,caspase-1 and GSDMD were assayed using immunoblotting.Caspase-1 activity in the liver tissues was determined with colorimetric measurement and immunofluorescence assay.Results Compared with HFD-fed WT mice,MYKI mice with HFD feeding showed milder liver function damage(P<0.01),alleviated hepatic steatosis,and reduced liver macrophage infiltration,glucose tolerance impairment and insulin resistance(P<0.01).The levels of IL-6,IL-1β,and TNF-α and mRNA expressions of M1 type macrophage markers were significantly decreased(P<0.01)and those of M2 type markers increased(P<0.01)in the liver tissues of HFD-fed MYKI mice,which also showed reduced NLRC4 inflammasome activity,caspase-1 activation,and GSDMD-N protein expression compared with their WT counterparts(P<0.05).Conclusion Overexpression of HEM2M reduces the production of hepatic inflammatory factors,improves insulin resistance and inhibits hepatic NLRC4 inflammasome activation,which leads to reduced hepatic pyroptosis and liver injury in NAFLD mice.

Objective The aim is to utilize CRISPR/Cas9 gene editing technology to construct Dmd gene mutant mice with a point mutation in exon 23 of the Dmd gene. Subsequently, the phenotypic changes of the mice in muscles and immune systems are analyzed and verified, providing an evaluation model for Duchenne muscular dystrophy and other related diseases.MethodsBased on the sequence characteristics of exon 23 of the Dmd gene, small guide RNA (sgRNA) was designed and synthesized. Cas9 mRNA, sgRNA fragments, and oligo donor DNA were microinjected into fertilized eggs of C57BL/6J mice. After transferring the fertilized eggs to surrogate mice, F0 generation mice were born. After mating with F0 generation mice, offspring mice were obtained, and Dmd gene positive mutant (Dmd^Mu/+) mice were obtained after genotype identification. Male hemizygous Dmd^Mu/+(Dmd^Mu/Y) mice were selected for phenotype validation. The body weight of live 3- and 9-month-old mice were recorded. Muscle tension was evaluated through the grid test. Hearts and semitendinosus muscles were collected, and the histopathological changes were observed using HE staining. Further, the expression of Dmd protein in muscle tissue of 9-month-old mice was analyzed by Western blotting.An acute inflammation model was established in Dmd^Mu/Y mice using lipopolysaccharide induction. Peripheral blood from the submandibular vein was collected, and the changes in the proportion of neutrophils and monocytes were detected by flow cytometry.Results The results of genome sequencing and Western blotting confirmed the successful construction of Dmd gene point mutant mice (Dmd^Mu/+ mice). Dmd protein expression was not detected in skeletal muscle and myocardium of Dmd^Mu/+ mice, and it was significantly reduced compared to wild-type C57BL/6J mice (P<0.05). Compared with wild-type mice of the same background, Dmd^Mu/Y mice at 3 and 9 months of age showed significant weight loss (P<0.01) and decreased muscle tension (P<0.05). 9-month-old Dmd^Mu/Y mice exhibited significant pathological changes in skeletal muscle and myocardium, including widening of intermuscular space. Under normal condition, compared with wild-type mice, the proportion of neutrophils and monocytes in the peripheral blood of 3-month-old Dmd^Mu/Y mice was significantly lower than that of wild-type mice (P<0.01). After lipopolysaccharide stimulation, the proportion of neutrophils in peripheral blood of 3-month-old Dmd^Mu/Y mice remained significantly lower compared to that of wild-type mice (P<0.01). The proportion of neutrophils in peripheral blood of 9-month-old Dmd^Mu/Y mice significantly decreased after lipopolysaccharide induction (P<0.01), with a trend of change observed in monocytes between groups.Conclusion The successful construction of the Dmd gene mutant mouse model has confirmed the vital function of Dmd gene in maintaining normal muscle tissue morphology and muscle tone. It preliminarily indicated that Dmd gene deletion could significantly reduce the proportion of neutrophils in peripheral blood, offering a new perspective for the study of immune system alterations in Duchenne muscular dystrophy patients.

OBJECTIVE To explore in vitro dissolution and in vivo pharmacokinetics of Luteolin solid dispersion in Beagle dogs. METHODS The dissolution of Luteolin solid dispersion was investigated according to the second method （paddle method） of the “dissolution determination method” in the 2020 edition of Chinese Pharmacopoeia （Part Ⅳ）. UPLC-MS/MS method was established to determine the concentration of luteolin in the plasma of Beagle dogs. Twelve Beagle dogs were randomly divided into luteolin group and Luteolin solid dispersion group， with 6 dogs in each group. They were given relevant medicine orally at the dose of 10 mg/kg luteolin. Blood was collected before medication （0 h）， at 5， 10， 15， 30， 45 min and 1， 2， 4， 6， 8， 10， 12， 24， 48 h after administration. After protein precipitation with acetonitrile， the blood concentration of luteolin in Beagle dogs was determined by UPLC-MS/MS and the major pharmacokinetic parameters were calculated with non-compartmental models by using DAS 3.2.8 pharmacokinetic software. RESULTS The dissolutions of Luteolin solid dispersion in purified water and 0.1% sodium dodecyl sulfate solution was significantly higher than those of luteolin； the dissolution rate reached 95% in 0.1% sodium dodecyl sulfate solution for 120 minutes. The peak concentration （cmax） of luteolin in the Luteolin solid dispersion group of Beagle dogs was 5.62 times higher than the luteolin group， and the relative bioavailability was 348%. Compared with luteolin group， cmax and the area under the drug time curve of luteolin in the Luteolin solid dispersion group of Beagle dogs were significantly increased， while the apparent distribution volume was significantly reduced （P＜0.05）. CONCLUSIONS Luteolin solid dispersion can improve in vitro dissolution and bioavailability of luteolin in Beagle dogs.

Objective To explore the effects of long non-coding RNA(lncRNA)HEM2M overexpression on liver injury in mice with non-alcoholic fatty liver disease(NAFLD).Methods Wild-type C57BL/6(WT)mice and myeloid cell-specific HEM2M knock-in(MYKI)mice were fed normal(ND)or high-fat diet(HFD)for 12 weeks.After intraperitoneal glucose tolerance and insulin tolerance tests,the mice were euthanized for detection of liver function indicators in the serum and liver tissue.HE staining and F4/80 immunohistochemical staining were used to examine liver pathologies,and the levels of IL-6,IL-1β,and TNF-α in the liver tissues were determined with ELISA.The mRNA expressions of HEM2M and the markers of M1 macrophages(TNF-α,iNOS,and IL-6)and M2 macrophages(Arg-1,YM-1,and IL-10)were detected using qRT-PCR,and the protein expressions of P-AKT,T-AKT,NLRC4,caspase-1 and GSDMD were assayed using immunoblotting.Caspase-1 activity in the liver tissues was determined with colorimetric measurement and immunofluorescence assay.Results Compared with HFD-fed WT mice,MYKI mice with HFD feeding showed milder liver function damage(P<0.01),alleviated hepatic steatosis,and reduced liver macrophage infiltration,glucose tolerance impairment and insulin resistance(P<0.01).The levels of IL-6,IL-1β,and TNF-α and mRNA expressions of M1 type macrophage markers were significantly decreased(P<0.01)and those of M2 type markers increased(P<0.01)in the liver tissues of HFD-fed MYKI mice,which also showed reduced NLRC4 inflammasome activity,caspase-1 activation,and GSDMD-N protein expression compared with their WT counterparts(P<0.05).Conclusion Overexpression of HEM2M reduces the production of hepatic inflammatory factors,improves insulin resistance and inhibits hepatic NLRC4 inflammasome activation,which leads to reduced hepatic pyroptosis and liver injury in NAFLD mice.

Objective We investigated the effects of Xueguan Ruanhua Pills(XGRHW) on ferroptosis in ApoE-/- atherosclerotic mice through the nuclear factor E2 related factor 2 (Nrf2)/xCT/glutathione peroxidase 4 (GPX4) signaling pathway.Methods Ten male C57BL/6J mice in the normal group were fed normal chow. Additionally, 50 ApoE-/- mice were fed high-fat chow for 12 weeks, and were divided into the following five groups (10 mice per group): the model group, the XGRHW low-dose (2.34g/kg) group, the XGRHW high-dose (4.68 g/kg) group, the XGRHW high-dose combined with the Nrf2 inhibitor ML385 (0.03 g/kg) group, and the ferrostatin-1 (1 mg/kg) group. Drugs were administered for 6 weeks. The blood levels of four types of lipids were detected by an automatic lipid analyzer, lipid deposition in the aorta was observed by Oil Red O staining, histomorphological changes in the aortic sinus were observed by HE staining, the serum levels of Fe2+, MDA, GSH, and SOD were determined by colorimetric assays, and the expression levels of FTH1 and FTL in the aortic sinus were observed by immunofluorescence. The protein levels of Nrf2, xCT, and GPX4 in mouse aortic tissues were detected by Western blotting. The ultrastructural changes of aortic mitochondria were observed by transmission electron microscopy.Results Compared with the normal group, mice in the model group showed obvious lipid plaque deposition in the aorta, severely calcified lesions in the aortic sinus, elevated serum levels of TC, TG, LDL-C, Fe2+, and MDA, decreased levels of HDL-C, SOD, and GSH (P<0.01), and decreased protein expressions of aortic Nrf2, xCT, and GPX4 as well as the iron storage proteins FTH1 and FTL (P<0.01), and serve damage to mitochondrial structure and morphology. Compared with the model group, the relative aortic plaque area was decreased, calcified lesions in the aortic sinus were decreased, serum levels of TC, TG, LDL-C, Fe2+, and MDA were decreased, and HDL-C, SOD, and GSH levels were increased in the XGRHW low-dose and high-dose and ferrostatin-1 groups (P<0.05 or P<0.01), and Nrf2, xCT, GPX4, and the iron storage proteins FTH1 and FTL were upregulated in aortic tissues (P<0.05 or P<0.01), and mitochondrial structure approaching normal. In the XGRHW high-dose+ML385 group, compared with the XGRHW high-dose group, the levels of blood lipids and lipid peroxidation were increased and the protein levels of Nrf2, xCT, and GPX4 in aortic tissue and the iron storage proteins FTH1 and FTL were decreased (P<0.01), and mitochondrial structure was damaged indicating that ML385 could inhibit the therapeutic effect of the XGRHW in atherosclerotic mice.Conclusion The XGRHW can improve blood lipid levels and reduce the degree of arterial plaque lesions in atherosclerotic mice, and its mechanism of action may be related to activation of the Nrf2/xCT/GPX4 pathway to inhibit ferroptosis.

Objective To construct a model of Fat-1/APPPS1 transgenic mice and a cellular model of microglia and explore the improvement effect and underlying mechanism of n-3 polyunsaturated fatty acids(n-3 PUFAs)on the learning and memory abilities of APPPS1 mice by regulating microglial activation and polarization.Methods After the male mice with heterozygous Fat-1 genotype were mated with the female ones with heterozygous APPPS1 genotype,genetic identification was used to screen the male offspring with Fat-1/APPPS1 genotype.Then after the male wild-type(WT)mice and those with Fat-1,Fat-1/APPPS1,and APPPS1 genotypes were bred until 9 months old,their learning and memory abilities were evaluated with Morris water maze(MWM)test.In addition,gas chromatography-mass spectrometry(GC-MS)was performed to detect the concentration of PUFAs in the brain,and immunohistochemistry(IHC)was applied to detect the deposition of β-amyloid protein(Aβ)in the hippocampal regions.Moreover,immunofluorescence assay,qRT-PCR,and enzyme-linked immunosorbent assays(ELISA)were conducted to measure inflammation,and transcription and expression of Iba-1(indicating the microglial activation)and CD86 and CD206(indicating microglial polarization)in central nervous system(CNS).After pretreated with DHA+EPA(25 pmol/mL∶25 μmol/mL),microglial model of inflammatory injury was established in immortalized mouse BV2 cells induced by LPS(1 μg/mL).Afterwards,immunofluorescence assay,qRT-PCR and Western blotting were used to detect inflammation and microglial activation and polarization.Results Compared with the APPPS1 mice,endogenous n-3 PUFAs effectively improved the learning and memory disorders in Fat-1/APPPS1 ones(P＜0.05),remarkably alleviated Aβ deposition in the hippocampal regions(P＜0.05),evidently reduced CNS inflammation and microglial activation(P＜0.05)and transformed the activated microglia from M1 to M2(P＜0.05).Furthermore,BV2 cells with DHA+EPA pretreatment obviously resisted LPS-induced cellular inflammation and induced activated ones from M1 to M2(P＜0.05).Conclusion n-3 PUFAs inhibit the microglial activation,regulate the microglial polarization from M1 to M2,reduce CNS inflammation,and thus alleviate learning and memory disorders in APPPS1 mice.

OBJECTIVE To explore the clinical characteristic manifestation of Rosai-Dorfman disease(RDD)involving the nasal cavity and paranasal sinuses.METHODS The clinical data of 16 patients with RDD involving the nasal cavity and paranasal sinuses who received treatment in Department of Rhinology of Beijing Tongren Hospital Affiliated to Capital Medical University from December 2016 to December 2023,were retrospective analyzed.RESULTS The male to female ratio was 1:3,with an average age of 47.4 years and an average disease duration of 22.34 months.There were 14,3,5,6 and 1 patients who complained of nasal congestion,dry nose,decreased sense of smell,head and face pain,and bulging/diplopia,respectively.There were 9 patients who had external nasal swelling.There were 14 RDD patients who were involved at least the nasal septum,12 patients who were simply affected in the nasal cavity,and 3 cases who involved the paranasal sinuses.There was 1 case belonged to a mixed type of RDD.There were 8 of 10 cases who were misdiagnosed or missed due to intraoperative rapid freezing.Endoscopic examination showed bilateral bulging in 9 patients'nasal septum,and nodular or granular new growth on 7 patients'nasal floor and inferior turbinate surface.Sinus CT found 9 patients'the nasal septum area showed a circular uniform soft tissue shadow or symmetrical soft tissue thickening shadow,5 patients'nasal floor and inferior turbinate showed obvious soft tissue thickening shadow.CONCLUSION RDD involving the nasal cavity and paranasal sinuses has certain typical features in external nasal manifestations,endoscopic and imaging examinations.Comprehensive judgment can help improve the feasibility of clinical diagnosis of RDD.