Metabolic-associated fatty liver disease (MAFLD) and osteoporosis (OP) are two very common metabolic diseases. A growing body of experimental evidence supports a pathophysiological link between MAFLD and OP. MAFLD is often associated with the development of OP. Rutaecarpine (RUT) is one of the main active components of Chinese medicine Euodiae Fructus. Our previous studies have demonstrated that RUT has lipid-lowering, anti-inflammatory and anti-atherosclerotic effects, and can improve the OP of rats. However, whether RUT can improve both fatty liver and OP symptoms of MAFLD mice at the same time remains to be investigated. In this study, we used C57BL/6 mice fed a high-fat diet (HFD) for 4 months to construct a MAFLD model, and gave the mice a low dose (5 mg·kg^-1) and a high dose (15 mg·kg^-1) of RUT by gavage for 4 weeks. The effects of RUT on liver steatosis and bone metabolism were then evaluated at the end of the experiment [this experiment was approved by the Experimental Animal Ethics Committee of Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences (approval number: IMB-20190124D₃03)]. The results showed that RUT treatment significantly reduced hepatic steatosis and lipid accumulation, and significantly reduced bone loss and promoted bone formation. In summary, this study shows that RUT has an effect of improving fatty liver and OP in MAFLD mice.

Objective Mobile artificial lumbar complex(MALC)which suitable for reconstruction after subtotal lumbar resection in goats was developed,and to test stability of the complex and postoperative lumbar segmental motor function.Methods Eighteen male boer goats aged from 1 to 2 years old(weighted from 35 to 45 kg)were selected and divided into con-trol group,fusion group and non-fusion group,with 6 goats in each group.According to preoperative CT scans and MRI exami-nations of lumbar,the goat MALC was designed and performed by 3D printed for non-fusion group.Operation was performed on three groups respectively,and only vertebral body and disc were exposed in control group.In fusion group,L4 part of vertebral body and the upper and lower complete disc tissues were removed,and the lumbar spine bone plate fixation was performed with titanium mesh bone grafting.In non-fusion group,vertebral body and disc were removed in the same way,and MALC was im-planted.AP and lateral X-rays of lumbar vertebrae in goat were taken at 6 months after surgery,in order to understand whether the plant was dislocated,displaced and fractured.Biomechanical tests were performed on the specimens by mechanical instru-ment to measure range of motion(ROM)of L2,3,L,4,L4,5intervertebral space and the overall ROM of L2-5 lumbar vertebrae.Results MALC of lumbar vertebra was designed by 3D printing,and its component artificial vertebrae and upper and lower ar-tificial end plates were manufactured.The semi-spherical structure was fabricated by precision lathe using high-crosslinked polyethylene material,and the prosthesis was assembled.Postoperative AP and lateral X-rays of lumbar vertebra at 6 months showed the implant position of implant and MALC were good without displacement and dislocation.In vitro biomechanical test of lumbar vertebrae specimens:(1)There were no statistical significance in ROM of lumbar intervertebral space flexion and extension,lateral flexion and rotation on L.4 and L4,5,between non-fusion group and control group(P＞0.05),while ROM of fu-sion group was significantly reduced compared with the other two groups(P＜0.05).There were no significant difference in ROM of L2.3 intervertebral flexion and extension,lateral flexion and rotation between non-fusion group and control group(P＞0.05),while fusion group was significantly increased compared with the other two groups(P＜0.001).(2)There was no signifi-cant difference in overall lumbar ROM of L2-5(P＞0.05).Conclusion The individual MALC could restore intervertebral height of lumbar vertebra while maintaining the stability of lumbar vertebra and re-establishing motor function of lumbar space.

This study aimed to determine the drug resistance phenotype and genetic characteristics of Listeria monocytogenes ST5 LK100 isolated from a neonate,which provided a basis for the diagnosis and treatment of L.monocyto-genes infection and to enhance the understanding of the genomic characteristics of this strain.A suspected L.monocytogenes strain was isolated from the gastric juice sample of an infected neonate,and identified with a VITEK2 Compact automatic mi-crobial identification instrument and 16S RNA sequencing.Five drug sensitivity tests were conducted on the identified strain with the E-test method.Additionally,the whole genome of the strain was sequenced using a third-generation sequencing plat-form.The antibiotic resistance elements of the strain were identified by BlastN with the CARD antibiotic resistance gene data-base.The multilocus sequence typing(MLST),serotyping,and virulence genes of the strain was determined by Pasteur da-tabase,the virulence gene distribution was analyzed using the virulence analysis website.The prophages of the strain were predicted and annotate by PHASTER online website.The strain(LK100)isolated from the neonate was identified as L.monocytogenes.This strain was sensitive to penicillin,ampicil-lin,meropenem,erythromycin,and trimethoprim-sulfame-thoxazole antibiotics.The MLST type and serotype was ST5 and 1/2b-3b,respectively.The total length of the chromoso-mal genome of LK100 was 3 032 582 bp with a GC content of 37.91％,and it contained a complete circular plasmid with a se-quence length of 52 822 bp.The strain LK100 carried complete InlA protein,LIPI-1 pathogenicity island,SSI-1 stress survival island,and an LGI2 genomic island.The intrinsic antibiotic resistance genes were mainly located on the chromosome.Five prophage sequences were predicted in the LK100 genome.This study identified a strain of ST5 L.monocytogenes LK100 from an infected neonate and characterized its genome and antibiotic sensitivity,laying the foundation for further research on ST5 L.monocytogenes.

Pancreatic polypeptide(PP),a pancreatic hormone containing 36 amino acids,plays impor-tant roles in the diagnosis and evaluation of pancreatic function,injury and diseases.In this study,a phage nanobody library against PP was constructed to screen specific PP nanobodies,which would be used to evaluate whether they have binding activity with PP antigen.After PP antigen with high purity was prepared by prokaryotic expression system,it was used to immunize alpaca to construct the nanobody li-brary against PP with high storage capacity and high abundance,from which 8 strains of PP nanobodies were obtained by phage display.One of nanobody strain(PP-VHH)was selected to be expressed in a prokaryotic expression system,which was induced overnight by IPTG.After purification and identifica-tion,the antigen-antibody binding activity and PP level in serum were detected by indirect ELISA and Sandwich ELISA methods,respectively.The results showed that PP-VHH had binding activity with PP,which could be used to detect PP in chicken and human serum.The Sandwich ELISA methods with R2 of the fitting curve 0.9868 could be used to detect PP concentrations of 48-55 pg/mL in the serum of chick-ens,while the concentrations of PP in human serum varied significantly.In summary,PP-VHH screened from nanobody library against PP could detect PP in serum,which would supply the basis for evaluation of abnormal pancreatic function and diagnosis of relative disease.

Alzheimer's disease(AD)is a neurodegenerative disease with age-related cognitive decline.Sphingosine-1-phosphate receptor 2(S1PR2)is involved in a variety of cellular processes and has been shown to play an important role in nervous system development.This study aimed to investigate the effects and possible mechanism of S1PR2 on Aβ25-35 induced cell model damage of AD.In this study,SH-SY5Y cells were induced by Aβ25-35 to construct a cell damage model,and the expression of S1PR2 in cells was interfered by targeting sequence.The protein and gene expression levels of S1PR2 were detected by Western blot and RT-PCR.It was found that the expression of S1PR2 was significantly increased at mR-NA and protein levels in Aβ25-35-induced SH-SY5Y cell model(P＜0.01),and its expression was signifi-cantly decreased after S1PR2 intervention(P＜0.001).The cell proliferation activity was detected by CCK8,and apoptosis was detected by flow cytometry.The results showed that the proliferation activity of Aβ25-35 induced SH-SY5Y cells was significantly increased,and apoptosis was decreased after S1PR2 in-tervention(P＜0.01).The protein levels of APP,Tau,p-Tau,and PSD95 in cells were detected by Western blot to analyze the effect of S1PR2 on the pathology of AD.It was found that after S1PR2 inter-vention,the expressions of APP,Tau,and p-Tau in the AD cell model were significantly decreased(P＜0.001),and the expression of synaptic protein PSD95 was increased(P＜0.001),which could signifi-cantly improve the pathological damage in Aβ25-35-induced SH-SY5Y cell model.In addition,ATP pro-duction was detected by the kit,and ROS content and mitochondrial membrane potential were detected by flow cytometry to analyze the mitochondrial function.Results found that ATP production and mitochondri-al membrane potential was significantly decreased,whereas the ROS content was increased in Aβ25 35 in-duced SH-SY5Y cells(P＜0.001).Intervention with S1PR2 significantly increased ATP production and mitochondrial membrane potential,but decreased ROS content(P＜0.001).Finally,the protein levels of the AKT/mTOR pathway were detected by Western blot.The results showed that S1PR2 significantly in-hibited the activation of the AKT/mTOR pathway induced by Aβ25-35 in SH-SY5Y cells.In conclusion,S1PR2 may be involved in the pathogenesis of Aβ25-35-induced SH-SY5Y cells by promoting mitochondrial function through the AKT/mTOR pathway.

Objective:To analyze the relationship between serum cystatin C(CysC),β2-microglobulin(β2-MG)and the efficacy of demethylation therapy in patients with acute myeloid leukemia(AML).Methods:A prospective cohort study was conducted on 98 AML patients admitted to the Affiliated Hospital of Inner Mongolia Medical University from February 2019 to January 2022.All patients were treated with decitabine(DAC)+HAG regimen,28 days as a course and treated for 3-4 courses.At the end of each course of treatment,the treatment effect of the patients was evaluated,and the patients who achieved complete remission(CR)transferred to consolidation therapy,while the patients who did not reach CR at the end of the course of treatment were considered as treatment failure.The examination items before treatment include routine blood parameters,serum CysC,and β2-MG,and general clinical data of the patients were collected.According to the statistical results,logistic regression model was used to analyze the relationship between serum CysC,β2-MG and the efficacy of demethylation therapy in AML patients.The ROC curves were drawn,and the predictive efficacy of serum CysC,β2-MG on demethylation therapy in AML patients was evaluated by the area under the curve(AUC).Results:Of the 98 AML patients enrolled in the study,5 cases were excluded during the treatment period,and 93 cases finally completed the chemotherapy courses.Among them,23 patients achieved CR after the initial induction chemotherapy(course 1-2),and 11 patients achieved CR after the re-induction chemotherapy(course 3-4).The success rate of demethylation therapy was 36.56％(34/93).Compared with the patients in treatment success group,patients in treatment failure group had a higher proportion of intermediate-and adverse-risk,lower levels of platelet(PLT)and hemoglobin(Hb),and higher expression levels of serum CysC and β2-MG,all of which were statistically significant(P＜0.05).Logistic regression analysis showed that high expression of serum CysC,β2-MG and adverse-risk were independent risk factors for failure of demethylation treatment in AML patients(OR＞1,P＜0.05).The ROC curves showed that the AUC values of serum CysC,β2-MG alone and combined in predicting the efficacy of demethylation therapy in AML patients were 0.788,0.785 and 0.834,respectively.Conclusion:The failure of demethylation therapy in AML patients is related to the high expression of serum CysC and β2-MG,and detection of serum CysC and β2-MG before treatment can predict the risk of demethylation therapy failure in AML patients.

Objective:To investigate the causal correlation between depression and stress urinary incontinence(SUI)using Mendelian randomization(MR)analysis.Methods:We searched the FinnGen Consortium database for genome-wide association studies(GWAS)on depression and obtained 23 424 case samples and 192 220 control samples,with the GWAS data on SUI provided by the UK Biobank,including 4 340 case samples and 458 670 control samples.We investigated the correlation between depression and SUI based on the depression data collected from the Psychiatric Genomics Consortium(PGC).We employed inverse-variance weighting as the main method for the MR study,and performed sensitivity analysis to verify the accuracy and stability of the findings.Results:Analysis of the data from the UK Biobank and FinnGen Consortium showed that depression was significantly correlated with an increased risk of SUI(P=0.005),but not SUI with the risk of depression(P=0.927).And analysis of the PGC data verified the correlation of depression with the increased risk of SUI(P=0.043).Conclusion:Depression is associated with an increased risk of SUI,while SUI does not increase the risk of depression.

AIM To study the chemical constituents from the leaves of Citrus reticulata Blanco and their anti-inflammatory activities.METHODS The 85%ethanol extract from the leaves of C.reticulata was isolated and purified by silica gel,D101 macroporous resin,MCI,ODS and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The Griess method was used to determine their inhibitory activities on lipopolysaccharide-induced NO production in macrophages RAW 264.7 cells.The mice foot swelling inflammation model induced by carrageenan was established,and the levels of IL-1β,TNF-α were detected.RESULTS Twelve compounds were isolated and identified as nobiletin(1),tangeretin(2),5-demethylinoblitin(3),5,4'-dihydroxy-6,7,8,3'-tetramethoxy flavone(4),5-hydroxy-7,8,3',4'-trimethoxyflavanone(5),3,5,6,7,8,3',4'-heptamethoxyflavanone(6),hesperetin(7),5-hydroxy-6,7,3',4'-tetramethoxyphenone(8),β-balsam alcohol(9),stigmaster-5-en-3β-alcohol(10),p-hydroxybenzaldehyde(11),vanillin(12).Compounds 1,4,6,7,10 and 12 had strong inhibitory activites on NO release in LPS-induced RAW 264.7 cells,and the IC50 values were(25.21±2.10),(37.77±0.50),(38.19±1.58),(21.89±1.73),(43.81±1.18),(47.98±2.55),(41.23±1.11),(43.80±1.43)μmol/mL,respectively.Compounds 2-3 reduced IL-1β and TNF-α levels(P＜0.05,P＜0.01).CONCLUSION Compounds 6-7,9 are isolated from this plant for the first time.Compounds 1-4,8 exhibit strong in vitro anti-inflammatory activities,and compounds 2-3 exhibit significant in vivo anti-inflammatory activities.

Objective To explore the changes in gut microbiota and levels of short-chain fatty acids(SCFA)in infants with cow's milk protein allergy(CMPA),and to clarify their role in CMPA.Methods A total of 25 infants diagnosed with CMPA at Children's Hospital Affiliated to Zhengzhou University from August 2019 to August 2020 were enrolled as the CMPA group,and 25 healthy infants were selected as the control group.Fecal samples(200 mg)were collected from both groups and subjected to 16S rDNA high-throughput sequencing technology and liquid chromatography-mass spectrometry to analyze the changes in gut microbial composition and metabolites.Microbial diversity was analyzed in conjunction with metabolites.Results Compared to the control group,the CMPA group showed altered gut microbial structure and significantly increased α-diversity(P＜0.001).The abundance of Firmicutes,Clostridiales and Bacteroidetes was significantly decreased,while the abundance of Sphingomonadaceae,Clostridiaceae_l and Mycoplasmataceae was significantly increased in the CMPA group compared to the control group(P＜0.001).Metabolomic analysis revealed reduced levels of acetic acid,butyric acid,and isovaleric acid in the CMPA group compared to the control group,and the levels of the metabolites were positively correlated with the abundance of SCFA-producing bacteria such as Faecalibacterium and Roseburia(P＜0.05).Conclusions CMPA infants have alterations in gut microbial structure,increased microbial diversity,and decreased levels of SCFA,which may contribute to increased intestinal inflammation.[Chinese Journal of Contemporary Pediatrics,2024,26(3):236-243]

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.