Objective:To investigate the role of WW domain containing E3 ubiquitin protein ligase 1 (WWP1) in enamel development of mice.Methods:Single-cell RNA sequencing data of incisor tissues of postnatal day 7 (P7) mice and mandibular first molar tooth germs of P3.5 mice were used to analyze the expression of Wwp1 in dental epithelial cells. Immunohistochemistry was performed to observe the distribution and expression levels of WWP1 in the epithelium of mouse incisors and mandibular first molar tooth germs. Wwp1 knockout (Wwp1 KO) mice were generated and collected with their control littermates at P1, P7, three mice per group, as well as at P14, P28, 2 months (2M), and 3M, six mice per group. The enamel volumes of molars and incisors were analyzed using micro-CT. Scanning electron microscopy was employed to examine the enamel cross-sections of Wwp1 KO and control mice. Energy dispersive spectroscopy (EDS) was used to analyze the calcium and phosphorus content of the enamel rod of incisors. Immunofluorescence was performed to detect the expression of amelogenin (AMELX) in the ameloblasts of Wwp1 KO and control mice. Additionally, LS-8 ameloblast-like epithelial cells were cultured, and Wwp1 siRNA or overexpression plasmids were transfected to knock down or overexpress WWP1. The protein levels of AMELX were then assessed by Western blotting.Results:Single-cell sequencing result showed a high Wwp1 mRNA expression level in the epithelial cells of mouse incisors and mandibular molar tooth germs. Immunohistochemistry revealed the expression of WWP1 in presecretory, secretory, transitional, and mature ameloblasts. Wwp1 KO mice exhibited enamel developmental defects. The enamel volumes of molars and incisors in Wwp1 KO mice [(0.155±0.016), (0.300±0.017) μm 3] were reduced by 23.95% ( P<0.001) and 28.31% ( P<0.001) compared with the control group [(0.203±0.062), (0.418±0.023) μm 3] respectively. Scanning electron microscopy showed disorganized enamel structures in Wwp1 KO incisors and molars. EDS results showed the weight percent of calcium in the enamel rod of incisors decreased in Wwp1 KO mice [(20.74±0.91)%] compared with the control group [(30.30±3.83)%] ( P<0.001), and the calcium-to-phosphorus ratio decreased in Wwp1 KO mice (1.93±0.01) compared with the control group (2.02±0.01) ( P<0.001). Immunofluorescence showed weaker AMELX expression in ameloblasts of mandibular first molar tooth germs from P1 and P7 Wwp1 KO mice compared with the control group ( P<0.001, P<0.001). In LS-8 cells, Wwp1 knocked-down led to a decrease of AMELX protein expression, while WWP1 overexpression resulted in an increased AMELX protein level. Conclusions:WWP1 promotes ameloblast differentiation and enamel matrix mineralization, playing a critical role in enamel formation.

Objective To investigate the application of transvaginal ultrasound(TVS),multimo-dal magnetic resonance imaging(MRI)and 18F-fluorodeoxyglucose positron emission computed tomo-graphy/computed tomography(18F-FDG PET/CT)in diagnosis of endometrial carcinoma(EC).Methods A total of 157 patients with endometrial cancer in our hospital were selected.The results of EC staging by multimodal MRI and transvaginal ultrasound and positive 18F-FDG PET/CT were recor-ded,and their results were compared with pathological results.Results The results of TVS showed 87 cases in stage Ⅰ,6 cases were misdiagnosed and 12 cases were missed diagnosis.In 55 cases of stage Ⅱ,12 cases were misdiagnosed and 8 cases were missed.In 15 cases of stage Ⅲ,2 cases were misdiagnosed without missed diagnosis.The results of multimodal MRI showed that 86 cases were mis-diagnosed in stage Ⅰ,3 cases were misdiagnosed and 10 cases were missed.In 58 cases of stage Ⅱ,10 cases were misdiagnosed and 3 cases missed diagnosis.There were 13 cases of stage Ⅲ with no missed diagnosis or misdiagnosis.The results of 18F-FDG PET/CT were positive in 145 cases and missed in 12 cases.Based on the golden standard of postoperative pathological staging,the accuracy of TVS di-agnosis of EC patients in stage Ⅰ was 87.10％(81/93),stage Ⅱ was 84.31％(43/51),and stage Ⅲ was 100％(13/13).Based on the golden standard of postoperative pathological staging,the accuracy of mul-timodal MRI in the diagnosis of EC patients was 89.25％(83/93)in stage Ⅰ,94.12％(48/51)in stageⅡ,and 100％(13/13)in stage Ⅲ.According to the pathological staging,145 EC positive cases were de-tected by 18F-FDG PET/CT,the accuracy rate was 92.36％(145/157).Conclusion 18F-FDG PET/CT has a high diagnostic accuracy in EC patients,TVS and multimodal MRI have a high value in the preoperative staging diagnosis of EC,and are worthy of clinical promotion.

Objective:To investigate the role of WW domain containing E3 ubiquitin protein ligase 1 (WWP1) in enamel development of mice.Methods:Single-cell RNA sequencing data of incisor tissues of postnatal day 7 (P7) mice and mandibular first molar tooth germs of P3.5 mice were used to analyze the expression of Wwp1 in dental epithelial cells. Immunohistochemistry was performed to observe the distribution and expression levels of WWP1 in the epithelium of mouse incisors and mandibular first molar tooth germs. Wwp1 knockout (Wwp1 KO) mice were generated and collected with their control littermates at P1, P7, three mice per group, as well as at P14, P28, 2 months (2M), and 3M, six mice per group. The enamel volumes of molars and incisors were analyzed using micro-CT. Scanning electron microscopy was employed to examine the enamel cross-sections of Wwp1 KO and control mice. Energy dispersive spectroscopy (EDS) was used to analyze the calcium and phosphorus content of the enamel rod of incisors. Immunofluorescence was performed to detect the expression of amelogenin (AMELX) in the ameloblasts of Wwp1 KO and control mice. Additionally, LS-8 ameloblast-like epithelial cells were cultured, and Wwp1 siRNA or overexpression plasmids were transfected to knock down or overexpress WWP1. The protein levels of AMELX were then assessed by Western blotting.Results:Single-cell sequencing result showed a high Wwp1 mRNA expression level in the epithelial cells of mouse incisors and mandibular molar tooth germs. Immunohistochemistry revealed the expression of WWP1 in presecretory, secretory, transitional, and mature ameloblasts. Wwp1 KO mice exhibited enamel developmental defects. The enamel volumes of molars and incisors in Wwp1 KO mice [(0.155±0.016), (0.300±0.017) μm 3] were reduced by 23.95% ( P<0.001) and 28.31% ( P<0.001) compared with the control group [(0.203±0.062), (0.418±0.023) μm 3] respectively. Scanning electron microscopy showed disorganized enamel structures in Wwp1 KO incisors and molars. EDS results showed the weight percent of calcium in the enamel rod of incisors decreased in Wwp1 KO mice [(20.74±0.91)%] compared with the control group [(30.30±3.83)%] ( P<0.001), and the calcium-to-phosphorus ratio decreased in Wwp1 KO mice (1.93±0.01) compared with the control group (2.02±0.01) ( P<0.001). Immunofluorescence showed weaker AMELX expression in ameloblasts of mandibular first molar tooth germs from P1 and P7 Wwp1 KO mice compared with the control group ( P<0.001, P<0.001). In LS-8 cells, Wwp1 knocked-down led to a decrease of AMELX protein expression, while WWP1 overexpression resulted in an increased AMELX protein level. Conclusions:WWP1 promotes ameloblast differentiation and enamel matrix mineralization, playing a critical role in enamel formation.

Thyroid-associated ophthalmopathy(TAO)is a rare organ-specific autoimmune disease with an unclear pathogenesis. At present, the treatment still relies mainly on glucocorticoids and traditional immunosuppressants. However, some patients respond poorly to these drugs and experience treatment-related adverse reactions, highlighting the urgent need for novel drugs for TAO treatment. In recent years, with the deepening of research on the pathogenesis of TAO, a multitude of biologics targeting specific targets have emerged. Among them, teprotumumab, which targets the insulin-like growth factor-I receptor(IGF-IR), has been approved by the Food and Drug Administration for the treatment of TAO, and several other biologics are currently in clinical trials. This review provides the latest reference for the clinical prevention, treatment, and research of TAO by summarizing the current clinical research status of biologics targeting IGF-IR, neonatal Fc receptor(FcRn), thyroid-stimulating hormone receptor(TSHR), B cells, cytokines, and other biological agents in TAO and analyzing their impact on clinical treatment and future research trends.

Thyroid-associated ophthalmopathy(TAO)is a multifactorial-mediated autoimmune orbital disease with the highest incidence of orbital disease in adults.Due to the complex clinical manifestations and prolonged course,TAO seriously affect the physical and mental health of patients.The pathogenesis of TAO has not been fully elucidated and the treatment lacks specificity.Therefore,in-depth research on the pathogenesis of TAO is to find effective treatments.In recent years,studies have suggested that there is gut microbiota disorder in TAO,and the risk factors of TAO can promote gut microbiota disorder.Disordered gut microbiota can participate in the occurrence and development of TAO via influencing T cell differentiation,mimicking autoantigens,and influencing host non-coding RNA expression.Modulating the gut microbiota also has therapeutic effects on TAO and is a promising therapeutic approach.

Objective:To re-evaluate the systematic reviews of the effectiveness of virtual reality technology in pain management in children.Methods:Using computers to search for systematic reviews of the effectiveness of virtual reality technology in pain management for pediatric patients in China National Knowledge Infrastructure (CNKI), Wanfang Database, VIP, China Biology Medicine disc, Cochrane Library, PubMed, Embase, CINAHL and Web of Science. The retrieval period was from the establishment of the databases to November 4, 2022. Literature screening and data extraction were conducted independently by two researchers, methodological quality evaluation was performed for the included studies using the A Measurement Tool to Assess Systematic Reviews 2 (AMSTAR 2) checklist, and the evidence quality rating was evaluated using Grades of Recommendations Assessment, Development and Evaluation (GRADE) system.Results:A total of 14 systematic reviews were included. AMSTAR 2 evaluation results showed that the methodological quality of 2 systematic reviews was low, and 12 were extremely low. GRADE was used to evaluate the quality of evidence of the outcome indicators, and the results showed that among the 26 outcome indicators included, 23.08% (6/26) were medium evidence, 76.92% (20/26) were low evidence, and the quality of no evidence was high.Conclusions:There is a lack of high-quality evidence to support the conclusion that virtual reality technology can relieve pain in children. The methodological quality and evidence quality of systematic reviews are low, and more standardized and rigorous studies need to be carried out in the future to further verify the application effect of virtual reality technology in pain management of children.

Objective: To analyze the clinical characteristics of SARS-CoV-2 Omicron variant infected children in convalescence in Tianjin. Methods: A total of 104 pediatric patients infected by SARS-CoV-2 Omicron variant Tianjin First Central Hospital (designated hospital for SARS-CoV-2 infection in Tianjin) for convalescent treatment from January 22^nd, 2022 to February 24^th were included for a retrospective study.Clinical data including clinical typing, SARS-CoV-2 IgG and IgM test and 2019-nCoV nucleic acid test were collected.The cases were divided into 2-dose group and zero-dose group based on the doses of inactivated SARS-CoV-2 vaccine. The children were divided into repositive group and negative group, according to the nucleic acid test during hospitalization. Chi-square test was used for the comparison between the groups. Results: The age of these 104 children was 10.0 (0.3, 14.0) years on admission, 53 children were males and 51 were females, 92 cases (88.5%) had mild symptoms, 12 cases (11.5%) had common symptoms.The age and SARS-CoV-2 IgG level of zero-dose group was lower (2.0 (0.3, 10.2) vs. 10.0 (3.2, 14.0) years, 10 (2, 17) vs. 193 (157, 215), χ²=-5.57, Z=-48.76,both P<0.001) than that of 2-dose group. The zero-dose group had a high rate of transmission among family members and a high level of SARS-CoV-2 IgM level (13/14 vs. 62.2% (56/90), 0.4 (0.2, 0.8) vs. 0.4 (0.2, 1.1),χ²=5.09, Z=-48.95, both P<0.05) than the 2-dose group. Repositive group had a high rate of underlying diseases and SARS-CoV-2 IgM level was higher (2/13) vs. 1.1% (1/91), (0.6 (0.2, 1.0) vs. 0.3 (0.2, 0.7), χ²=8.29, Z=2.70, both P<0.05) than negative group. The SARS-CoV-2 IgG level of repositive group was lower than that of negative group (160 (78, 197) vs. 213 (186, 231), χ²=-3.20, P<0.05). Conclusions: Children infected with SARS-CoV-2 Omicron variant in Tianjin were mainly transmitted by family members, and most of them had mild symptoms. Two-dose group had higher IgG levels and lower IgM levels than zero-dose group.The probability of SARS-CoV-2 nucleic acid test repositivity increased in children with underlying diseases and lower IgG levels.
Antibodies, Viral ; COVID-19/therapy* ; COVID-19 Vaccines ; Child ; Female ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Male ; Nucleic Acids ; Retrospective Studies ; SARS-CoV-2

Graves' ophthalmopathy is the most common clinical orbital disease, and T helper (Th) cells play an important role in the development of Graves' ophthalmopathy. Th17 cells are a major subpopulation of Th cells and abnormally highly expressed in patients with Graves' ophthalmopathy. Th17 cells and the related cytokines interleukin (IL)-17A, IL-21 and IL-23 are involved in regulating the inflammatory response, fibrosis and adipogenesis. Th17 cells are unstable and exhibit a degree of plasticity, and they can differentiate into IL-17A and interferon (IFN)-γ dual-producing Th17.1 cells, which exacerbate the pathogenicity of Th17 cells. In addition, Th17 cells and the relevant factors are strongly associated with disease activity and severity in Graves' ophthalmopathy.
Humans ; Cytokines ; Th17 Cells ; Graves Ophthalmopathy ; Adipogenesis

Objective:To explore the difference of high-risk factors between early-onset and late-onset pre-eclampsia, and to further understand high-risk factors of pre-eclampsia.Methods:Clinical data of pre-eclampsia pregnant women in 160 medical institutions in China in 2018 were retrospectively analyzed, including 8 031 cases of early-onset pre-eclampsia and 12 969 cases of late-onset pre-eclampsia. The proportion of high-risk factors, different body mass index (BMI) and age stratification between early-onset group and late-onset group were compared.Results:(1) Univariate analysis of high-risk factors: the proportions of high-risk factors in early-onset group and late-onset group were compared, and the differences were statistically significant (all P<0.05). Among them, the proportions of primipara and multiple pregnancy in early-onset group were lower than those in late-onset group, while the proportions of pregnant women with advanced age, irregular antenatal examination, obesity, family history of hypertension, pre-eclampsia, diabetes, kidney diseases, immune system diseases and assisted reproductive technology were higher than those in late-onset group. (2) Hierarchical analysis of BMI: the proportion of pregnant women with BMI≥24 kg/m 2 in early-onset group [48.2% (2 828/5 872) vs 45.5% (4 177/9 181), respectively; P=0.001] and the proportion of pregnant women with BMI ≥28 kg/m 2 in early-onset group [19.5% (1 143/5 872) vs 18.0% (1 656/9 181), respectively; P=0.028] were significantly higher than those in late-onset group. (3) Age stratification analysis: the proportion of pregnant women aged 35-39 years in the early-onset group [21.8% (1 748/8 023) vs 17.5% (2 110/12 068), respectively; P<0.01], the proportion of pregnant women 40-44 years old [6.8% (544/8 023) vs 5.4% (648/12 068), respectively; P<0.01], and the proportion of pregnant women ≥45 years old [0.7% (58/8 023) vs 0.5% (57/12 068), respectively; P=0.021] were significantly higher than those in the late-onset group. (4) Multivariate analysis: advanced age (≥35 years old), multiple pregnancy, irregular antenatal examination or transfer from other hospitals, family history of hypertension (including paternal, maternal and parental lines), previous history of pre-eclampsia, kidney diseases, immune system diseases (systemic lupus erythematosus, antiphospholipid antibody syndrome) and assisted reproductive technology pregnancy were the risk factors affecting the severity of pre-eclampsia (all P<0.05). Conclusion:Pregnant women with high risk factors such as age ≥35 years old, BMI ≥24 kg/m 2 before pregnancy, family history of hypertension, history of pre-eclampsia, chronic kidney diseases, immune diseases (mainly including systemic lupus erythematosus and antiphospholipid syndrome) and assisted reproductive technology are more likely to have early-onset pre-eclampsia.