Objective:To prepare pullulan-spermine (PS)-poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) conjugated with CD3 antibody, and to investigate their effects on T cell proliferation and cytokine secretion.Methods:Purulan polysaccharide was sperminized to synthesize PS, hydrophobically modified, and then grafted with PLGA to synthesize PS-PLGA. Infrared spectroscopy and nuclear magnetic resonance hydrogen spectrum were used to characterize the structure of PS-PLGA. PS-PLGA NPs were prepared by ultrasonic dialysis method and then coupled with CD3 antibody to prepare PS-PLGA-CD3 NPs. The morphological features of PS-PLGA-CD3 NPs were observed by the transmission electron microscope. The particle sizes, Zeta potential and dispersive coefficient of the NPs were measured using the dynamic laser particle size analyzer. The amount of coupled CD3 antibody on the surface of the NPs was determined using quantitative fluorescence analysis method. The effects of 1, 10, 50, 100, and 200 μg/ml PS-PLGA-CD3 NPs on T-cell proliferation were determined using cell counting kit-8 method. The effects of 1, 10, 50, 100, 200 μg/ml PS-PLGA-CD3 NPs on secretion of interferon-γ (IFN-γ), interleukin-2 (IL-2), and tumor necrosis factor-β (TNF-β) by T cell were determined by enzyme-linked immunosorbent assay. Comparisons were made using independent sample t-test or one-factor analysis of variance. Results:Pullulan and PS showed strong absorption at 2 939 cm ?1, and PS had a weaker absorption peak at 3 384 cm ?1 than pullulan. The proton peaks of spermine appeared at chemical shifts of 1.25 to 1.50, 1.63, and 2.25 to 2.75. The characteristic peaks of PLGA appeared at chemical shifts of 1.50, 3.40, and 4.80 to 5.30. Compared to pullulan, the characteristic peaks of both PS and PLGA appeared in the corresponding intervals for PS-PLGA. The morphology of PS-PLGA-CD3 NPs with spermine substitution at 9.7% was all regular and circular, with a mean particle size of (173.3±24.5) nm, a Zeta potential of (?12.78±3.68) mV, the dispersive coefficient of 0.254±0.101, and the CD3 antibody mass fraction of (52.1±9.4) μg/mg. The differences in cell survival were statistically significant for PS-PLGA-CD3 NPs and PS-PLGA NPs, respectively, after co-incubation with T cell after 24, 48, and 72 h at concentrations of 50, 100, and 200 μg/ml, respectively (all P<0.05). The results of the three concentration comparisons after 24 h of co-incubation were [(129.8±23.1)% vs (95.5±8.9)%, (137.5±22.7)% vs (95.1±15.8)%, and (142.3±25.6)% vs (93.2±9.2)%]; and the results after 48 h were [(145.9±23.7)% vs (95.8±10.6)%, (149.3±23.5)% vs (94.9±16.3)%, and (161.2±26.9)% vs (91.5±8.3)%]; and the results after 72 h were [(147.6±20.1)% vs (95.9±17.8)%, (152.4±22.3)% vs (92.7±16.5)%, and (167.7±25.4)% vs (90.8±17.4)%]. The differences in the levels of IFN-γ, IL-2 and TNF-β were statistically significant (all P<0.05 or 0.01) at 50, 100 and 200 μg/ml concentrations for PS-PLGA-CD3 NPs and PS-PLGA NPs, respectively. For IFN-γ, the results of the comparison of the three concentrations were [(35.7±3.1) ng/ml vs (16.4±6.9) ng/ml, (67.3±5.2) ng/ml vs (19.6±2.8) ng/ml, and (79.0±4.2) ng/ml vs (19.3±2.3) ng/ml]; and for IL-2, the results were [(43.5±8.2) ng/ml vs (12.6±1.9) ng/ml, (53.5±7.8) ng/ml vs (15.8±3.3) ng/ml, and (64.0±8.2) ng/ml vs (17.4±3.8) ng/ml]; and for TNF-β, the results were [(108.4±18.9) pg/ml vs (40.8±1.3) pg/ml, (152.3±28.3) pg/ml vs (56.4±3.7) pg/ml and (185.0±33.6) pg/ml vs (81.6±10.2) pg/ml]. Conclusions:PS-PLGA-CD3 NPs are successfully prepared, which have the function of effectively promoting T cell proliferation and cytokine sectetion.

Objective: To explore the association between dietary inflammatory index (DII) and metabolic syndrome (MetS) and its components among children aged 6-14 years in Beijing, so as to provide a reference for preventing MetS. Methods: A cross sectional study was carried out in 2 086 records of 1 832 children from the 2017 and 2019 Nutrition and Health Surveillance in Primary and Secondary school students of Beijing (NHSPSB). Three day consecutive 24 hour dietary recalls combined with weighing household cooking oils and condiments were used to collect dietary intake and calculate DII. MetS was diagnosed according to "Definition and Suggestion on the Metabolic Syndrome of Chinese Children and Adolescent". The Generalized estimating equations (GEEs) models were used to analyze the association between DII and the presence of MetS and its components (abdominal obesity, high triglyceride, low high density lipoprotein cholesterol, hypertension, and hyperglycemia). Results: The mean DII score was (1.64±1.07) for the included children. No significant association was found between DII scores and the likelihood of MetS (per 1 point increment: OR =1.16, 95% CI =0.92-1.48, P >0.05). In terms of the components of MetS, DII scores were positively associated with the odds of high triglyceride (per 1 point increment: OR =1.17, 95% CI =1.01-1.36, P <0.05). There was no statistically significant difference in the association among different age groups ( P >0.05). No significant associations were observed between DII and other MetS components( P >0.05). Conclusion DII scores may not be correlated with the risk of MetS, but proinflammatory diet might increase the risk of high triglyceride. DII score in childhood should be emphasized to identify and prevent MetS as soon as possible.

Model informed precision dosing for warfarin is to provide individualized dosing by integrating information related to patient characteristics, disease status and pharmacokinetics /pharmacodynamics of warfarin, through mathematical modeling and simulation techniques based on the quantitative pharmacology. Compared with empirical dosing, it can improve the safety, effectiveness, economy, and adherence of pharmacotherapy of warfarin. This consensus report describes the commonly used modeling and simulation techniques for warfarin, their application in developing and adjusting dosing regimens, medication adherence and economy. Moreover, this consensus also elaborates the detailed procedures for the implementation in the warfarin pharmacy service pathway to facilitate the development and application of model informed precision dosing for warfarin.

Model informed precision dosing (MIPD) is a new concept to guide precision dosing for individual patient by modeling and simulation based on the available information about the individual patient, medications and the disease. Compared to the empirical dosing, MIPD could improve the efficacy, safety, economics and adherence of the pharmacotherapy according to the individual's pathophysiology, genotyping and disease progression. This consensus report provides a brief account of the concept, methodology and implementation of MIPD as well as clinical decision supporting systems for MIPD. The status and future advancing of MIPD was also discussed to facilitate the appropriate application and development of MIPD in China.

A splicing mutation in VPS4B can cause dentin dysplasia type I (DD-I), a hereditary autosomal-dominant disorder characterized by rootless teeth, the etiology of which is genetically heterogeneous. In our study, dental follicle cells (DFCs) were isolated and cultured from a patient with DD-I and compared with those from an age-matched, healthy control. In a previous study, this DD-I patient was confirmed to have a loss-of-function splicing mutation in VPS4B (IVS7 + 46C > G). The results from this study showed that the isolated DFCs were vimentin-positive and CK14-negative, indicating that the isolated cells were derived from the mesenchyme. DFCs harboring the VPS4B mutation had a significantly higher proliferation rate from day 3 to day 8 than control DFCs, indicating that VPS4B is involved in cell proliferation. The cells were then replenished with osteogenic medium to investigate how the VPS4B mutation affected osteogenic differentiation. Induction of osteogenesis, detected by alizarin red and alkaline phosphatase staining in vitro, was decreased in the DFCs from the DD-I patient compared to the control DFCs. Furthermore, we also found that the VPS4B mutation in the DD-I patient downregulated the expression of osteoblast-related genes, such as ALP, BSP, OCN, RUNX2, and their encoded proteins. These outcomes confirmed that the DD-I-associated VPS4B mutation could decrease the capacity of DFCs to differentiate during the mineralization process and may also impair physiological root formation and bone remodeling. This might provide valuable insights and implications for exploring the pathological mechanisms underlying DD-I root development.
ATPases Associated with Diverse Cellular Activities ; genetics ; Case-Control Studies ; Cell Differentiation ; genetics ; Cells, Cultured ; Dental Sac ; cytology ; Dentin Dysplasia ; genetics ; pathology ; physiopathology ; Endosomal Sorting Complexes Required for Transport ; genetics ; Humans ; Mutation ; genetics ; Osteogenesis ; genetics ; RNA Splicing ; genetics

Objective:To observe the presence or absence of necroptosis in PC12 cells after radiation injury, and to detect the expression of receptor-interacting protein 3(RIP3) and evaluate its regulatory effect on necroptosis.Methods:PC12 cells were treated with different doses of irradiation and their necroptosis was detected by lactate dehydrogenase (LDH) release at different time points. After pretreatment with necroptosis inhibitor Necrostatin-1(Nec-1), the changes of cell necroptosis were detected by LDH. The expression level of RIP3 after irradiation intervention was detected by Western blot (WB). After pretreatment with the RIP3-specific inhibitor GSK′872, the changes of cell necroptosis were detected by LDH. The best transfection sequence of RIP3 knockout was screened by WB. The cells were divided into the control group, irradiation group, solvent control group, no-load control group and pretreatment group. WB, immunofluorescence staining, MTT, LDH and Annex V-fluorescein Isothiocyanate/Propidium Iodide (AnnexV-FITC/PI) flow cytometry were used for detection and analysis.Results:After 4 Gy irradiation, the degree of cell necrosis was the highest after 3 hours of culture, and the expression level of RIP3 protein was up-regulated. The cell necrosis was decreased after Nec-1, GSK′872 and RIP3 gene knockdown pretreatment.Conclusions:The radiation injury of 4 Gy can induce the necroptosis of PC12 cells, and the most significant effect can be observed when cultured for 3 hours after irradiation. RIP3 is involved in the process of necroptosis of PC12 cells induced by radiation injury, and plays a pivotal positive regulatory role.

Objective To investigate the clinical characteristics of polyarteritis nodosa (PAN) patients with renal involvement. Methods PAN patients admitted to the department of rheumatology, department of pediatrics, department of nephrology, general internal medicine department and department of vascular surgery at Peking Union Medical College Hospital from June 2012 to August 2018 were enrolled in this study and were divided into two groups according to renal involvement or not. The clinical characteristics were analyzed. Results A total of 94 PAN patients were finally enrolled and 57 (60.64%) presented kidney manifestation. The mean age of onset was (37.76±17.40) years old and the interval from onset to diagnosis was 10 (0 to 240) months. Forty patients were misdiagnosed once or more times. In patients with renal involvement, 9 cases suffered from renal ischemia or infarction, 31 with microscopic haematuria, 26 with proteinuria, renal artery or its branch involved in 17 cases, renal vein thrombosis in 1 case, 4 cases with pyeloureterectasis, one case with renal fascia thickening, 33 cases with impaired renal function (serum creatinine>84 μmol/L) including creatinine>140 μmol/L in 10 patients. Renal artery branch stenosis was the most common presentation [9 cases (52.94%)] of renal vascular involvement, other abnormalities including nodular dilatation [4 cases (23.53%)], occlusion [3 cases (17.65%)]. There were significant differences (P<0.05) in the PAN patients with and without renal involvement in the following: age of onset [(33.72±16.13) years vs. (43.97±17.66)years, t2=2.901, P=0.005], weight loss(≥4kg since PAN onset) [25(43.86%) vs. 7(18.92%), χ2=6.216, P=0.013], elevation of diastolic blood pressure [22(38.60%) vs. 7 (18.92%), χ2=4.072, P=0.044], acromegaly gangrene [18(31.58%) vs. 21(56.76%), χ2=5.859, P=0.015], and gastrointestinal artery involvement [20(35.09%) vs. 6(1.22%), χ2=3.993, P=0.046]. Laboratory parameters and the application of glucocorticoid and cyclophosphamide therapies were similar in two groups (all P>0.05). Conclusion Young PAN patients are more likely to be associated with renal involvement, especially gastrointestinal arteries.

Objective To investigate the clinical characteristics of gastrointestinal involvement in polyarteritis nodosa (PAN),and to improve the understanding of the disease.Methods PAN patients hospitalized in Peking Union Medical College Hospital from March 2002 to September 2016 were enrolled in this study,and were divided into gastrointestinal involvement group and non-gastrointestinal involvement group according to clinical manifestations and imaging findings.Data on clinical features,treatments and outcome were recorded.t test,chi-square test were used for statistical analysis.Results A total of 117 patients with PAN were hospitalized in the past 14 years.The prevalence of gastrointestinal involvement was 38％(44 cases).There was no significant difference in age and sex between the two groups (P＞0.05).Abdominal pain (29 cases,66％) was the most frequent manifestation,then gastrointestinal bleeding (10 cases,23％),splenic infarction (3 cases,7％),gastrointestinal ulcers (2 cases,5％),intestinal obstruction or diarrhea (each 2 cases,5％),and vomiting (1 case,2％).Patients with gastrointestinal involvement had more frequent fatigue (27％ vs 11％;x2=5.156,P=0.023),increased diastolic pressure (55％ vs 34％;x2=4.647,P=0.031),renal (34％ vs 18％;x2=3.998,P=0.046) and cardiac (25％ vs 8％;x2=6.225,P=0.013) involvements.ESR in the gastrointestinal involvement group was significantly higher (75％ vs 56％;x2=4.190,P=0.041).The average follow-up time was 315.8 (20.3,441.3) days,the relapse rate was higher in the gastrointestinal involvement group (23％ vs 8％;x2=4.895,P=0.027).The incidence of death or the irreversible organ injury was higher in the gastrointestinal involvement group (27％ vs 11％,x2=5.156,P=0.023).Conclusion Gastrointestinal invol-vement in poly-arteritis nodosa is common and its condition is severe.The incidence of relapse and death or irreversible organ injury is high.

Objective: To investigate the clinical characteristics of polyarteritis nodosa (PAN) patients with renal involvement. Methods: PAN patients admitted to the department of rheumatology, department of pediatrics, department of nephrology, general internal medicine department and department of vascular surgery at Peking Union Medical College Hospital from June 2012 to August 2018 were enrolled in this study and were divided into two groups according to renal involvement or not. The clinical characteristics were analyzed. Results: A total of 94 PAN patients were finally enrolled and 57 (60.64%) presented kidney manifestation. The mean age of onset was (37.76±17.40) years old and the interval from onset to diagnosis was 10 (0 to 240) months. Forty patients were misdiagnosed once or more times. In patients with renal involvement, 9 cases suffered from renal ischemia or infarction, 31 with microscopic haematuria, 26 with proteinuria, renal artery or its branch involved in 17 cases, renal vein thrombosis in 1 case, 4 cases with pyeloureterectasis, one case with renal fascia thickening, 33 cases with impaired renal function (serum creatinine>84 μmol/L) including creatinine>140 μmol/L in 10 patients. Renal artery branch stenosis was the most common presentation [9 cases (52.94%)] of renal vascular involvement, other abnormalities including nodular dilatation [4 cases (23.53%)], occlusion [3 cases (17.65%)]. There were significant differences (P<0.05) in the PAN patients with and without renal involvement in the following: age of onset [(33.72±16.13) years vs. (43.97±17.66) years, t²=2.901, P=0.005], weight loss(≥4kg since PAN onset) [25(43.86%) vs. 7(18.92%), χ²=6.216, P=0.013], elevation of diastolic blood pressure [22(38.60%) vs. 7(18.92%), χ²=4.072, P=0.044], acromegaly gangrene [18(31.58%) vs. 21(56.76%), χ²=5.859, P=0.015], and gastrointestinal artery involvement [20(35.09%) vs. 6(1.22%), χ²=3.993, P=0.046]. Laboratory parameters and the application of glucocorticoid and cyclophosphamide therapies were similar in two groups (all P>0.05). Conclusion Young PAN patients are more likely to be associated with renal involvement, especially gastrointestinal arteries.

OBJECTIVETo analyze the clinical phenotype of a Chinese pedigree affected with hereditary dentinogenesis imperfecta and mutation of dentin sialophosphoprotein (DSPP) gene.

METHODSAffected members underwent intraoral photography, dental film and panoramic radiography. Genomic DNA was extracted from peripheral venous blood samples. Coding regions of the DSPP gene were subjected to PCR amplification and Sanger sequencing. Functional effect of the mutation was predicted with SIFT and PolyPhen-2. The tertiary structure of wild type and mutant proteins were predicted by Swiss-Port.

RESULTSA heterozygous c.50C to T (p.P17L) mutation was identified in exon 2 of the DSPP gene in the proband and her father. The same mutation was not found among 200 unrelated healthy controls. The Pro-17 residues and its surrounding positions in DSPP are highly conserved across various species. The mutation was predicted to be damaging to the structure of DSPP protein.

CONCLUSIONThe c.50C to T (p.P17L) mutation of the DSPP gene probably underlies the disease in this pedigree. Above finding has expanded the spectrum of DSPP gene mutations and provided a basis for genetic counseling and prenatal diagnosis for this family.