OBJECTIVE: To identify the role of gene silencing or overexpression of Nemo-like kinase (NLK) during the process of neural differentiation of human mesenchymal stem cells (hBMSCs), and to explore the effect of NLK downregulation by transfection of small interfering RNA (siRNA) on promoting neuralized tissue engineered bone regeneration. METHODS: NLK-knockdown hBMSCs were established by transfection of siRNA (the experimental group was transfected with siRNA silencing the NLK gene, the control group was transfected with control siRNA and labeled as negative control group), and NLK-overexpression hBMSCs were established using lentivirus vector transfection technique (the experimental group was infected with lentivirus overexpressing the NLK gene, the control group was infected with an empty vector lentivirus and labeled as the empty vector group). After neurogenic induction, quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression of neural-related gene, and Western blot as well as immunofluorescence staining about several specific neural markers were used to evaluate the neural differentiation ability of hBMSCs.6-week-old male nude mice were divided into 4 groups: ① β-tricalcium phosphate (β-TCP) group, ② β-TCP+ osteogenic induced hBMSCs group, ③ β-TCP+ siRNA-negative control (siRNA-NC) transfection hBMSCs group, ④ β-TCP+ siRNA-NLK transfection hBMSCs group. Four weeks after the subcutaneous ectopic osteogenesis models were established, the osteogenesis and neurogenesis were detected by hematoxylin-eosin (HE) staining, Masson staining and tissue immunofluorescence assay. Statistical analysis was conducted by independent sample t test. RESULTS: After gene silencing of NLK by siRNA in hBMSCs, neural-related genes, including the class Ⅲ β-tubulin (TUBB3), microtubule association protein-2 (MAP2), soluble protein-100 (S100), nestin (NES), NG2 proteoglycan (NG2) and calcitonin gene-related peptide (CGRP), were increased significantly in NLK-knockdown hBMSCs compared with the negative control group(P < 0.05), and the expression levels of TUBB3 and MAP2 of the NLK silencing group were also increased. Oppositely, after NLK was overexpressed using lentivirus vector transfection technique, TUBB3, MAP2, S100 and NG2 were significantly decreased in NLK-overexpression hBMSCs compared with the empty vector group (P < 0.05), and the expression level of TUBB3 was also decreased. 4 weeks after the subcutaneous ectopic osteogenesis model was established, more mineralized tissues were formed in the β-TCP+ siRNA-NLK transfection hBMSCs group compared with the other three groups, and the expression of BMP2 and S100 was higher in the β-TCP+ siRNA-NLK transfection hBMSCs group than in the other groups. CONCLUSION Gene silencing of NLK by siRNA promoted the ability of neural differentiation of hBMSCs in vitro and promoted neuralized tissue engineered bone formation in subcutaneous ectopic osteogenic models in vivo in nude mice.
Bone Regeneration/genetics* ; Animals ; Mesenchymal Stem Cells/cytology* ; Humans ; RNA, Small Interfering/genetics* ; Tissue Engineering/methods* ; Cell Differentiation ; Mice, Nude ; Gene Silencing ; Mice ; Male ; Protein Serine-Threonine Kinases/genetics* ; Intracellular Signaling Peptides and Proteins/genetics* ; Transfection ; Cells, Cultured ; Lentivirus/genetics*

Objective A non-targeted metabolomics analysis of vitreous fluid from patients with proliferative diabetic retinopathy(PDR)is conducted to explore the"metabolic map"of PDR.This approach aims to deepen the understanding of the disease,identify potential biomarkers.Methods From 35 PDR patients and 30 fresh rhegmatogenous retinal de-tachment(RRD)patients,10 PDR patients with vitreoretinal abnormal adhesions were selected as the experimental group(PDR group),and 10 fresh RRD patients were chosen as the control group(RRD group).Using ultra-high-performance liq-uid chromatography-mass spectrometry non-targeted metabolomics technology,the metabolic profiles of vitreous fluid were analyzed to obtain metabolic spectra.One-dimensional and multidimensional statistical methods were used to analyze the differences in metabolites and metabolic pathways between the PDR and RRD groups.Results A total of 165 differential metabolites were identified in the vitreous humor samples of patients in the PDR and RRD groups,these differential metab-olites were significantly enriched in 21 metabolic pathways(P＜0.05),Among these pathways,those with at least 5 differ-ential metabolites include:methionine and cysteine metabolism;glycine,serine,and threonine metabolism;ascorbic acid and aldose metabolism;amino acid biosynthesis;and central carbon metabolism in cancer.Pyruvate,serine,D-2-phospho-glycerate,threonine,phosphoserine,and high serine are present in multiple metabolic pathways,the areas under the curve are 0.96,0.82,0.85,0.78,0.40,and 0.31,respectively.Conclusion There are 21 significantly different metabolic pathways between PDR and RRD patients.Pyruvate stands out in multiple pathways,potentially serving as a biomarker for PDR diagnosis.

Objective A non-targeted metabolomics analysis of vitreous fluid from patients with proliferative diabetic retinopathy(PDR)is conducted to explore the"metabolic map"of PDR.This approach aims to deepen the understanding of the disease,identify potential biomarkers.Methods From 35 PDR patients and 30 fresh rhegmatogenous retinal de-tachment(RRD)patients,10 PDR patients with vitreoretinal abnormal adhesions were selected as the experimental group(PDR group),and 10 fresh RRD patients were chosen as the control group(RRD group).Using ultra-high-performance liq-uid chromatography-mass spectrometry non-targeted metabolomics technology,the metabolic profiles of vitreous fluid were analyzed to obtain metabolic spectra.One-dimensional and multidimensional statistical methods were used to analyze the differences in metabolites and metabolic pathways between the PDR and RRD groups.Results A total of 165 differential metabolites were identified in the vitreous humor samples of patients in the PDR and RRD groups,these differential metab-olites were significantly enriched in 21 metabolic pathways(P＜0.05),Among these pathways,those with at least 5 differ-ential metabolites include:methionine and cysteine metabolism;glycine,serine,and threonine metabolism;ascorbic acid and aldose metabolism;amino acid biosynthesis;and central carbon metabolism in cancer.Pyruvate,serine,D-2-phospho-glycerate,threonine,phosphoserine,and high serine are present in multiple metabolic pathways,the areas under the curve are 0.96,0.82,0.85,0.78,0.40,and 0.31,respectively.Conclusion There are 21 significantly different metabolic pathways between PDR and RRD patients.Pyruvate stands out in multiple pathways,potentially serving as a biomarker for PDR diagnosis.

Objective:To quantitatively evaluate the accuracy of data obtained from liquid-interference surfaces using an intraoral 3D scanner(IOS)integrated with a compressed airflow system,so as to pro-vide clinical proof of accuracy for the application of the compressed airflow system-based scanning head in improving data quality on liquid-interference surfaces.Methods:The study selected a standard model as the scanning object,adhering to the"YY/T 1818-2022 Dental Science Intraoral Digital Impression Scanner"guidelines,a standard that defined parameters for intraoral scanning.To establish a baseline for accuracy,the ATOS Q 12M scanner,known for its high precision,was used to generate true reference values.These true values served as the benchmark for evaluating the IOS performance.Building on the design of an existing scanner,a new scanning head was developed to integrate with a compressed airflow system.This new design aimed to help the IOS capture high-precision data on sur-faces where liquid-interference,such as saliva,might otherwise degrade scanning accuracy.The tradi-tional scanning method,without airflow assistance,was employed as a control group for comparison.The study included five groups in total,one control group and four experimental groups,to investigate the effects of scanning lens obstruction,airflow presence,liquid media,and the use of the new scan-ning head on scanning process and accuracy.Each group underwent 15 scans,generating ample data for a robust statistical comparison.By evaluating trueness and precision in each group,the study as-sessed the impact of the compressed airflow system on the accuracy of IOS data collected from liquid-interference surfaces.Additionally,we selected Elite and Primescan scanners as references for numeri-cal accuracy values.Results:The scanning accuracy on liquid-interference surfaces was significantly reduced in terms of both trueness and precision[Trueness:18.5(6.5)vs.38.0(6.7),P＜0.05;Preci-sion:19.1(8.5)vs.31.7(15.0),P＜0.05].The use of the new scanning head assisted by the com-pressed airflow system significantly improved the scanning accuracy[Trueness:22.3(7.6)vs.38.0(6.7),P＜0.05;Precision:25.8(9.6)vs.31.7(15.0),P＜0.05].Conclusion:The scanning head based on the compressed airflow system can assist in improving the accuracy of data obtained from liquid-inter-ference surfaces by the IOS.

Objective:To investigate the survival rate and clinical failure reasons of onlay and occlusal veneer restorations retrospectively,and to put forward valuable suggestions for the selection of clinical in-dications.Methods:A total of 102 patients and 124 teeth treated by one of the authors from 2016 to 2019 were subjected to CAD/CAM lithium silicate reinforced glass-ceramic onlay or veneer restorations of premolars and molars,including 43 teeth with pulp vitality,81 endodontic treated teeth,and occlusal thickness of restoration was 1.5 mm.After four years of restoration,retrospective surveys were conducted to record the survival rate of restorations,the causes of restoration failure,and patient satisfaction rates,and the survival rate of restorations between vital teeth and endodontic treated teeth and among restored teeth was statistically analyzed by Chi-square test.Results:The survival rates of restorations on vital teeth and endodontic treated teeth were 95.5％and 90.0％,respectively,the average survival rate was 90.2％.The survival rates of vital teeth were higher than those of endodontic treated teeth without statis-tical difference.There was also no statistically significant difference among the tooth locations.The cau-ses of failure included the cracking of the restoration,the loss of the restoration,the fracture of the abut-ment teeth,secondary caries below the adjacent contact point,and food impaction caused by the loose-ning of the adjacent contact point.The overall patient satisfaction rate was 91.5％.Conclusion:The 4-year survival rate of glass-ceramic onlays and occlusal veneers is lower than that of the full crown restora-tion,and there are more complications than that of the single-crown restorations.The design of the resto-ration should be carefully selected based on the vitality of the abutment tooth and the remaining amount of tooth tissue.When there is too little tooth structure left,a post and crown should be selected for restora-tion.Adequate strength and thickness of the restoration should be ensured to prevent food impaction.Due to the small amount of abutment tooth preparation,it has the advantages of less stimulation of the pulp and periodontal tissue,and can be recommended as a trial restoration.

In fixed prosthodontics, clear exposure of the preparation margin is the prerequisite for obtaining accurate digital impressions and improving the marginal fit of restorations. To resolve the issues associated with the cord retraction technique, such as pain, acute injury, and prolonged procedural time, this study proposes a new technology for intraoral digital impression taking with pneumatic gingival retraction. The new scanning head blows a high-speed airflow that instantaneously separates the free gingiva, locally exposing the subgingival preparation margin. Combined with the farthest point preservation stitching algorithm based on the distance from the normal vector and high-speed laser scanning photography, it achieves global preparation edge data and gingival reconstruction, realizing painless, non-invasive, and efficient precise acquisition of the preparation margin. Using this new technique, a patient with a full porcelain crown restoration on a posterior tooth was treated. The digital impression revealed a clear margin of the preparation, and the crown made from this data has a good marginal fit.

To explore the differentially expressed genes (DEGs) related to biosynthesis of active ingredients in wolfberry fruits of different varieties of Lycium barbarum L. and reveal the molecular mechanism of the differences of active ingredients, we utilized Illumina NovaSeq 6000 high-throughput sequencing technology to conduct transcriptome sequencing on the fruits of 'Ningqi No.1' and 'Ningqi No.7' during the green fruit stage, color turning stage and maturity stage. Subsequently, we compared the profiles of related gene expression in the fruits of the two varieties at different development stages. The results showed that a total of 811 818 178 clean reads were obtained, resulting in 121.76 Gb of valid data. There were 2 827, 2 552 and 2 311 DEGs obtained during the green fruit stage, color turning stage and maturity stage of 'Ningqi No. 1' and 'Ningqi No. 7', respectively, among which 2 153, 2 050 and 1 825 genes were annotated in six databases, including gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and clusters of orthologous groups of proteins (KOG). In GO database, 1 307, 865 and 624 DEGs of green fruit stage, color turning stage and maturity stage were found to be enriched in biological processes, cell components and molecular functions, respectively. In the KEGG database, the DEGs at three developmental stages were mainly concentrated in metabolic pathways, biosynthesis of secondary metabolites and plant-pathogen interaction. In KOG database, 1 775, 1 751 and 1 541 DEGs were annotated at three developmental stages, respectively. Searching the annotated genes against the PubMed database revealed 18, 26 and 24 DEGs related to the synthesis of active ingredients were mined at the green fruit stage, color turning stage and maturity stage, respectively. These genes are involved in carotenoid, flavonoid, terpenoid, alkaloid, vitamin metabolic pathways, etc. Seven DEGs were verified by RT-qPCR, which showed consistent results with transcriptome sequencing. This study provides preliminary evidences for the differences in the content of active ingredients in different Lycium barbarum L. varieties from the transcriptional level. These evidences may facilitate further exploring the key genes for active ingredients biosynthesis in Lycium barbarum L. and analyzing their expression regulation mechanism.
Flavonoids/metabolism* ; Fruit/genetics* ; Gene Expression Profiling/methods* ; Gene Expression Regulation, Plant ; Lycium/metabolism* ; Metabolic Networks and Pathways ; Transcriptome

Objective:To evaluate the clinical survival rates and influence factors of different types of resin-bonded fixed partial dentures (RBFPD) used in anterior missing teeth restoration.Methods:Ninety-three RBFPD were delivered to 92 patients [92 patients,43 males and 49 females, average age (46.1±12.8) years] who visited Peking University School and Hospital of Stomatology from January 2006 to December 2021 for restoration of 1 or 2 anterior missing teeth. Altogether 32 cases of glass fiber reinforced RBFPD, 39 cases of glass-based ceramic RBFPD and 22 cases of porcelain-fused-to-metal RBFPD were retrospectively analyzed. The complete survival rate, functional survival rate, patients′ satisfaction and color matching of the restorations were recorded and evaluated every year since the replacement with RBFPD. The Kaplan-Meier survival curve method was used for survival analysis, and the Log-rank analysis was used to compare the effect of the number of missing teeth, position (maxillary or mandibular), cantilever or non-cantilever and gender on the survival rate of the restorations.Results:The overall survival time for the 93 RBFPD was 13.7 years (95% CI: 12.3-15.1 years). There was a decreasing trend in complete survival and functional survival for all three material RBFPD from year to year, but complete and functional survival rates exceeded 90% at year 5 and exceeded 80% at year 10. The complete survival rate of the glass-ceramic RBFPD was higher than the other two during the follow-up period, with a complete survival rate of 90% (35/39) at year 15. The porcelain-fused-to-metal RBFPD had a higher functional survival rate in years 1-8, but the complete and functional survival rates showed a substantial decrease after year 9. The single-factor Log-rank analysis showed that the success rate of porcelain-fused-to-metal RBFPD was significantly higher than that of glass fiber reinforced RBFPD (χ2=7.33, P=0.007), and the success rate of RBFPD with 1 missing tooth restored was significantly higher than that of RBFPD with 2 missing teeth restored (χ2=3.23, P=0.072). The differences in success rates between different restoration positions (maxillary and mandibular), cantilever or non-cantilever, and gender factors were not statistically significant (χ2=2.26, P=0.133; χ2=0.68, P=0.411; χ2=1.07, P=0.300). Conclusions:For the restoration of individual missing anterior teeth, both porcelain-fused-to-metal RBFPD and glass-based ceramic RBFPD achieve a high long-term clinical success rate, with glass-based ceramic RBFPD being more able to ensure long-term restorative result.

Autoimmune or infectious diseases often instigate the undesirable damages to tissues or organs to trigger immune-related diseases, which involve plenty of immune cells, pathogens and autoantibodies. Nanomedicine has a great potential in modulating immune system. Particularly, biomimetic nanomodulators can be designed for prevention, diagnosis and therapy to achieve a better targeted immunotherapy. With the development of materials science and bioengineering, a wide range of membrane-coated nanomodulators are available. Herein, we summarize recent advancements of bioinspired membrane-coated nanoplatform for systemic protection against immune-related diseases including autoimmune and infectious diseases. We also rethink the challenges or limitations in the progress of the therapeutic nanoplatform, and discuss the further application of the nanomodulators in the view of translational medicine for combating immune-related diseases.

OBJECTIVE：To systematically evaluate the effectiveness and safety of classical prescription versus antihistamines in the treatment of eczema. METHODS ：Retrieved from CBM and CNKI ，randomized controlled trials （RCTs）about classical prescription（trial group ）versus antihistamines （control group ）in the treatment of eczema were collected. The qualities of included literatures were evaluated by modified Jadad scale after literature screening and data extraction ，and Meta-analysis was performed by using Rev Man 5.3 software. RESULTS ：Totally 19 RCTs involving 1 829 patients were included. Meta-analysis showed that total response rate of patients with Shire Yunfu type [OR ＝5.67，95％CI（3.44，9.33），P＜0.000 01]，Pixu Shiyun type [OR ＝4.23， 95％ CI（1.89，9.46），P＝0.000 4]，Xuexu Fengzao type [OR ＝3.57，95％ CI（2.36，5.41），P＜0.000 01] in trial group were significantly higher than that of control group. There were no statistical differences in the incidence of ADR between patients with Shire Yunfu type [OR ＝0.72，95％CI（0.21，2.41），P＝0.59]，Pixu Shiyun type [OR ＝0.31，95％CI（0.01，13.57），P＝0.54]，Xuexu Fengzao type [OR＝1.08，95％ CI（0.18，6.59），P＝0.93] in trial group and control group. After treatment ，the levels of TNF-α [SMD＝－1.67，95％CI（－2.04，－1.31），P＜0.000 01]，IL-6 [SMD＝－1.53，95％CI（－1.88，－1.17），P＜0.000 01] and IL-8 [SMD＝－1.75，95％CI（－2.28，－1.21），P＜0.000 01] in patients with Shire Yunfu type in trial group were significantly lower than control group. There was no statistical significance in CD 4+ levels [SMD ＝1.51，95％CI（－0.37，3.38），P＝0.12] between Shire Yunfu type patients in trial group and control group. CONCLUSIONS ：Therapeutic efficacy of classical prescription in treatment of eczema is better than antihistamines，with similar safety . Modified Longdan xiegan decoction can significantly reduce the levels of inflammatory cytokines in patients with Shire Yunfu type.