Uncovering the risk factors of pulmonary hypertension and its mechanisms is crucial for the prevention and treatment of the disease.In the current study,we showed that experimental periodontitis,which was established by ligation of molars followed by orally smearing subgingival plaques from patients with periodontitis,exacerbated hypoxia-induced pulmonary hypertension in mice.Mechanistically,periodontitis dysregulated the pulmonary microbiota by promoting ectopic colonization and enrichment of oral bacteria in the lungs,contributing to pulmonary infiltration of interferon gamma positive(IFNγ+)T cells and aggravating the progression of pulmonary hypertension.In addition,we identified Prevotella zoogleoformans as the critical periodontitis-associated bacterium driving the exacerbation of pulmonary hypertension by periodontitis,and the exacerbation was potently ameliorated by both cervical lymph node excision and IFNγ neutralizing antibodies.Our study suggests a proof of concept that the combined prevention and treatment of periodontitis and pulmonary hypertension are necessary.

The proportion of overweight children within the obese population cannot be disregarded.Furthermore，obese children are susceptible to complications that may persist into adulthood.In order to prevent childhood obesity and manage its associated complications，it is essential to conduct appropriate assessments.In comparison to the body mass index（BMI）assessments，body composition analysis can measure the amount of adipose tissue，muscle tissue，body water，and inorganic salts.It can also assess body fat percentage，body muscle percentage，and muscle-fat ratio，which may be crucial in assessing body shape，basal metabolism，obesity and more.This enables accurate assessment and intervention.Bioelectric impedance analysis（BIA）is widely used in pediatrics as a method of analyzing body composition.However，its value in the study of obese children remains uncertain.This paper aims to summarize the value of BIA in the diagnosis and assessment of obese children and to assess its potential for screening for complications associated with childhood obesity，and reviews the performance of BIA in the assessment of obese children.

Objective:To investigate the impact of small extracellular vesicles(sEVs) derived from perirenal adipose cells on the biological behavior of renal tubular epithelial cells under diabetic conditions and the underlying molecular mechanisms.Methods:Primary perirenal adipose cells were extracted from db/m and db/db mice for in vitro culture. The culture supernatant was collected and sEVs(NDM-sEVs PRAT-Adipo, DM-sEVs PRAT-Adipo) were extracted by ultracentrifugation. The sEVs were incubated with human renal tubular epithelial cell line(HK-2) to observe changes in their proliferation, apoptosis, autophagy, and epithelial-mesenchymal transition(EMT) levels. The protein composition of sEVs was analyzed using mass spectrometry to explore the molecular mechanisms. Results:CCK8 results showed that the proliferation level of HK-2 cells after DM-sEVs PRAT-Adipo intervention did not change significantly compared with the two control groups(Ctrl group and NDM-sEVs PRAT-Adipo intervention group). Western Blot(WB) results indicated that there were no significant changes in apoptosis levels(Bcl-2, Cleaved-caspase 3, Caspase 3) and autophagy levels(p62, LC3BⅠ, LC3BⅡ) in the DM-sEVs PRAT-Adipo intervention group compared with the two control groups. WB and immunofluorescence results demonstrated that DM-sEVs PRAT-Adipo intervention upregulated the expression levels of mesenchymal cell marker proteins(Vimentin, α-SMA, Snail2) and downregulated the expression level of epithelial cell marker protein ZO-1 in HK-2 cells compared with the two control groups. Mass spectrometry analysis of sEVs revealed that the differential proteins between DM-sEVs PRAT-Adipo and NDM-sEVs PRAT-Adipo were enriched in EMT-related pathways. Among them, the enrichment of thrombospondin(THBS1) in DM-sEVs PRAT-Adipo might be involved in the regulation of EMT in HK-2 cells. Conclusion:Under diabetic conditions, sEVs secreted by PRAT-derived adipocytes promote the upregulation of EMT in renal tubular epithelial cells, a process that may be mediated by the enrichment of THBS1 in sEVs.

Objective:To explore the characteristics of key ferroptosis-related genes as therapeutic targets for sepsis based on bioinformatics analysis, and describe their immune characteristics.Methods:The transcriptome datasets GSE57065, GSE9960, GSE28750, and GSE137340 were downloaded from the Gene Expression Omnibus (GEO) database, immune-related gene (IRG) were obtained from ImmPort and InnateDB databases, and ferroptosis-related gene (FRG) were downloaded from the FerrDb database. The datasets GSE57065, GSE9960, and GSE28750 were integrated into an analysis dataset by the surrogate variable analysis (SVA) package and analyzed this analysis dataset by using the "limma" package to obtain differentially expressed gene (DEG), then the intersection set of DEG, FRG, and IRG were considered as ferroptosis and immune-related DEG (FImDEG). Gene ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using "ClusterProfiler" to understand the biological function of FImDEG. The key genes were screened by protein-protein interaction (PPI) network, least absolute shrinkage and selection operator (LASSO) regression algorithms, and support vector machine (SVM) analyses, and Logistic regression model was built based on above key genes. Receiver operator characteristics curve (ROC curve) was plotted to evaluate the diagnostic efficacy of the key genes alone or combinative. The degree of infiltration of 22 immune cells was assessed using the "CIBERSORT" package, and the correlation between the expressions of key genes and infiltration degree of immune cells was analyzed. Dataset GSE137340 was used to verify these key genes.Results:A dataset consisting of 146 sepsis samples and 61 healthy control samples was obtained by processing the database and removing batch effect. A total of 4?537 DEG were obtained, including 2?066 up-regulated genes and 2?471 down-regulated genes. 2?519 IRG and 855 FRG were obtained from the relevant database. Using the intersection of DEG, IRG and FRG, 34 FImDEG were obtained, including 20 up-regulated genes and 14 down-regulated genes. GO functional annotation showed that the biological functions of 34 FImDEG were mainly inhibition of transferase activity, regulation of DNA-binding transcription factor activity and cell response to stimulation. In terms of molecular function, it was mainly related to RNA polymerase Ⅱ-specific DNA-binding transcription factor binding and various protein ligase binding. Changes in cell composition occurred mainly in promyelocytic leukemia protein and chromatin silencing complexes. Enrichment analysis of KEGG pathway showed that the major pathways involved in 34 FImDEG included cell aging, expression of programmed death-ligand 1 (PD-L1) and programmed death-1 (PD-1) checkpoint pathways in cancer, interleukin-17 (IL-17) signaling pathway, lipid and atherosclerosis, and NOD-like receptor signaling pathway. Four key genes, including cytochrome b-245 β chain (CYBB), mitogen-activated protein kinase 14 (MAPK14), prostaglandin-endoperoxide synthase 2 (PTGS2) and V-relreticuloendotheliosis viral oncogene homology A (RELA), were screened through PPI network and LASSO and SVM machine learning. ROC curve analysis showed that the area under ROC curve (AUC) of the four key genes for diagnosing sepsis was all greater than 0.65, and the AUC of MAPK14 was 0.911. Logistic regression model was constructed based on four key genes, and the AUC was 0.956. Immunoinfiltration analysis showed that compared with healthy control samples, the infiltration degree of neutrophils and macrophages M0 was significantly increased in sepsis samples, while the infiltration degree of resting natural killer cell (NK cell), naive CD4 + T cell and CD8 + T cell was significantly lowered. Correlation analysis showed that the positive correlation between MAPK14 expression and the infiltration degree of neutrophils was the highest. Validation results in the GSE137340 dataset showed that compared with healthy control samples, the expressions of CYBB and MAPK14 in sepsis samples were significantly up-regulated, however, the expressions of PTGS2 and RELA were significantly down-regulated, similar to the expression trend in the above analysis dataset. Conclusion:Four key genes, including CYBB, MAPK14, PTGS2, and RELA, in the development of sepsis were identified through bioinformatics analysis, which play an important role in the immune process, and MAPK14 may be an important target for immune intervention.

Objective: Investigate osteogenic differentiation of canine periodontal ligament stem cells ( cPDLSCs) via over-expression ephrinB2 in cPDLSCs． Methods : cPDLSCs were isolated from the premolars and molars of Beagle．After transfected with EfnB2-GFP-Bsd and GFP-Bsd empty Vector，cPDLSCs were induced to osteogenic differentiation．Western blot was used to invest the expression of ephrinB2 protein．The effect of osteogenic differentiation of EfnB2-cPDLSCs and Vector-cPDLSCs were analyzed by RT-PCR , CCK-8，Alizarin-red S staining and ALP. Results: There was no significant difference in cell proliferation between EfnB2-cPDLSCs and Vector-cPDLSCs．While EfnB2-cPDLSCs displayed an enhanced ALP activity and more prominent mineralized nodules compared with Vector-cPDLSCs．The odonto-/ osteogenic genes in EfnB2-cPDLSCs were also highly enhanced． Conclusion The results of our study indicated that ephrinB2 gene-transfected cPDLSCs showed enhanced osteogenic differentiation.

Objective:To investigate the incidence rate and gene variation of methylmalonic academia (MMA) in Ji′nan city by analyzing biochemical and genetic screening results, and to explore the carrier frequency of MMA-related pathogenic genes in the population in Ji′nan.Methods:The children diagnosed with MMA by tandem mass spectrometry screening in Ji′nan Neonatal Disease Screening Centre from May 2011 to May 2022 were enrolled in this study.Their genetic test results were retrospectively analyzed and summarized.The dried heel blood tablets collected from 6 800 newborns were tested for neonatal gene screening. MMAA, MMAB, MMACHC and MMUT genes in 4 800 cases were detected by high-throughput sequencing+ target area capture technology.Ultra-multiplex polymerase chain reaction+ target gene locus capture technology was used to detect 174 target loci of 8 genes related to MMA in 2 000 cases.The hotspot mutation and related gene carrier rate of MMA were analyzed. Results:A total of 367 452 newborns were screened by tandem mass spectrometry, and 103 cases (56 males and 47 females) were diagnosed with MMA by screening.The estimated incidence of MMA was 1∶3 567.Among the 103 MMA cases, 76 were genetically diagnosed, and 4 gene variants of MMA ( MMAHC, MMUT, MMAA, MMADHC) were identified.A total of 6 800 neonates underwent neonatal genetic screening.Three of them were diagnosed with MMA.About 318 infants carried pathogenic variants of MMA, with a total carrier rate of 4.68%.Specifically, the carrier rates of MMACHC and MMUT gene variants were 3.09%(210/6 800) and 1.43% (97/6 800), respectively. Conclusions:MMA is the most common organic acid metabolism disorder in our country.The incidence and carrier rate of this disease are high in Jinan city.Neonatal genetic screening is an important supplement to neonatal biochemical screening.Carrier screening for MMA-related pathogenic genes is recommended for couples of childbearing age in Jinan.

Recent studies have found that the brain-gut-microbiome axis(BGMA)is closely related to the occurrence and development of Alzheimer's disease(AD). BGMA can affect AD in various aspects such as neuro-immune regulation and intestinal microflora, and is a potential new target for the treatment of AD.The "Sanjiao" acupuncture method is proposed by professor Han Jingxian, a famous Chinese medicine practitioner, based on his theory of "dysfunction of Qi activity of Sanjiao leads to aging" , and has been widely used in the treatment of AD and other age-related diseases in clinical practice.This article reviews the theory of "dysfunction of Qi activity of Sanjiao leads to aging" and the relationship between the "Sanjiao" acupuncture method and BGMA, with the hope that the "San Jiao" acupuncture method can become a new target for treatment of AD in the future.

OBJECTIVES: To retrospectively analyze the variation and characteristics of phenylalanine hydroxylase (PAH) gene, and to observe the long-term treatment effect and follow-up of newborns with PAH deficiency. METHODS: Clinical data, treatment and follow-up results of 198 patients with PAH deficiency diagnosed by newborn screening in Jinan from 1996 to 2021 were collected. The genetic analysis of 55 patients with PAH deficiency diagnosed by newborn screening in Jinan and 213 patients referred from the surrounding areas of Jinan were summarized. Gene variations were checked by a customized Panel gene detection method. Blood phenylalanine-concentration and physical development indicators including height and weight were regularly monitored. Intellectual development was assessed using a neuropsychological development scale for patients aged 0-6 years and academic performance, and brain injury in patients was assessed using brain magnetic resonance imaging. RESULTS: c.728G>A, c.158G>A, c.721C>T, c.1068C>A, c.611A>G variations were common in PAH gene. The genotype of c.158G>A variation is compound heterozygous variation, with mainly a mild hyperpheny-lalaninemia. 168 patients with PAH deficiency who were followed-up regularly had normal physical development without dwarfism or malnutrition. Among the 33 preschool patients who underwent mental development assessment, 2 were mentally retarded and the initial treatment age was older than 6 months. Nine patients with an average age of (17.13±2.42) years completed brain magnetic resonance imaging, one case was normal, and 8 cases were abnormal. There were patchy or patchy hyperintense foci near the bilateral lateral ventricles on T₂WI, and the intellectual development was normal. Compared with the other eight patients, the blood phenylalanine concentration of the normal child was better and stably controlled within the ideal range. CONCLUSIONS c.728G>A, c.158G>A, c.721C>T, c.1068C>A, c.611A>G variations were common in PAH gene. After standardized treatment, most patients with PAH deficiency diagnosed by screening can obtain normal growth and intellectual development in adolescence, but there are different degrees of organic lesions in the cerebral white matter.
Child ; Child, Preschool ; Adolescent ; Humans ; Infant, Newborn ; Young Adult ; Adult ; Neonatal Screening ; Follow-Up Studies ; Retrospective Studies ; Phenylketonurias/genetics* ; Phenylalanine Hydroxylase/genetics* ; Phenylalanine/therapeutic use* ; Mutation

Although widely applied in treating hematopoietic malignancies, transplantation of hematopoietic stem/progenitor cells (HSPCs) is impeded by HSPC shortage. Whether circulating HSPCs (cHSPCs) in steady-state blood could be used as an alternative source remains largely elusive. Here we develop a three-dimensional culture system (3DCS) including arginine, glycine, aspartate, and a series of factors. Fourteen-day culture of peripheral blood mononuclear cells (PBMNCs) in 3DCS led to 125- and 70-fold increase of the frequency and number of CD34⁺ cells. Further, 3DCS-expanded cHSPCs exhibited the similar reconstitution rate compared to CD34⁺ HSPCs in bone marrow. Mechanistically, 3DCS fabricated an immunomodulatory niche, secreting cytokines as TNF to support cHSPC survival and proliferation. Finally, 3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization. Our 3DCS successfully expands rare cHSPCs, providing an alternative source for the HSPC therapy, particularly for the patients/donors who have failed in HSPC mobilization.
Antigens, CD34/metabolism* ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Leukocytes, Mononuclear/metabolism* ; Peptides/metabolism*

Objective:To investigate the current situation of postcholecystectomy syndrome (PCS) and analyze its influencing factors, so as to provide basis for the formulation of postoperative nursing measures.Methods:A total of 336 patients who underwent laparoscopic cholecystectomy in Department of General Surgery in Xuanwu Hospital, Capital Medical University from June 2019 to December 2020 were selected as research objects by the convenient sampling method. General information questionnaire, lifestyle questionnaire, Self-rating Anxiety Scale and Self-rating Depression Scale were used to conduct a questionnaire survey of the patients in 1 month and 3 months after the operation. Logistic regression analysis was used to explore the influencing factors of PCS occurrence.Results:After 3 months of follow-up, 4 patients were lost to follow-up, and data of 332 patients were finally collected. The incidence of PCS was 23.5% (78/332) in 1 month after surgery and 7.2% (24/332) in 3 months after surgery. The total incidence of PCS within 3 months was 30.7% (102/332) , and the main symptoms were diarrhea, abdominal pain and abdominal distension. Logistic regression analysis showed that meat diet and irregular meals were the risk factors for PCS in one month after surgery ( P<0.05) and marriage was the protective factor for PCS in one month after surgery ( P<0.05) . Anxiety was a risk factor for PCS in 3 months after surgery ( P<0.05) . Conclusions:Patients with cholecystectomy have a high incidence of PCS. Patients are advised to reduce the intake of high-fat food, pay attention to the combination of meat and vegetables in their diet and correct irregular eating habits within 1 month after surgery. Meanwhile, family support should be enhanced and emotional management should be strengthened in 3 months after surgery to reduce the incidence of PCS.