AIM: To evaluate the diagnostic value of serum Th1-type cytokine levels and extraocular muscle-related parameters in thyroid-associated ophthalmopathy(TAO).METHODS: A retrospective study was conducted on 45 patients diagnosed with TAO in our hospital from January 2023 to December 2024, and 20 normal volunteers during the same period as controls. Venous blood samples of the patients were collected to detect the concentrations of Th1-type cytokines [interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α), interleukin-2(IL-2), and interleukin-12(IL-12)] in the serum. Additionally, the end diastolic velocity(Ved), velocity maximum(Vmax), resistance index(RI)of the central retinal artery, as well as the thickness and left-right diameter of the medial rectus muscle were measured. Logistic regression model was used to analyze the risk factors of TAO, and receiver operating characteristic curve(ROC)was adopted to evaluate the diagnostic efficacy of each index for the occurrence of TAO.RESULTS: The general information of the two groups was comparable. Compared with the normal control group, the serum concentrations of IFN-γ and TNF-α in TAO patients were significantly increased, Ved and Vmax were lower than those in the control group, and RI and the thickness of the medial rectus muscle were higher than those in the control group(all P<0.05). Logistic regression analysis showed that serum IFN-γ concentration, Ved, Vmax, and the thickness of the medial rectus muscle were all risk factors for TAO. ROC curve analysis indicated that the AUCs of serum IFN-γ concentration, Ved, Vmax, and the thickness of the medial rectus muscle for the diagnostic efficacy of TAO were 0.756, 0.769, 0.732, and 0.642, respectively. The combined detection of IFN-γ, Ved, Vmax, and the thickness of the medial rectus muscle had an AUC of 0.840 and a Youden index of 0.59, which was superior to the detection of a single indicator.CONCLUSION: The levels of serum Th1-type cytokines and extraocular muscle-related ultrasound indicators have certain value in the diagnosis of TAO. The combination of IFN-γ, Ved, Vmax, and the thickness of the medial rectus muscle has better diagnosis efficiency in TAO, which can provide a certain reference for the early diagnosis of TAO.

Objective To establish a sandwich enzyme-linked immunosorbent assay (ELISA) method for the quantitative detection of Chromogranin A (CHGA) in saliva, and to explore its preliminary application in the testing of saliva samples. Methods Recombinant human CHGA protein was used to immunize BALB/c mice, and monoclonal antibodies (mAbs) were prepared and screened using conventional hybridoma technology. A double-antibody sandwich ELISA detection method was constructed, and the matrix effect of saliva samples was optimized. This method was then applied to detect the concentration of CHGA in the saliva of stressed individuals. Results Twenty-one stable hybridoma cell lines secreting high affinity anti-human CHGA antibodies were obtained. A pair of detection antibodies with the best effect was selected, and the optimal coating concentration was determined to be 10 μg/mL, with the optimal dilution of detection antibodies being 1:32 000. The accuracy and reproducibility of this method were verified, with both intra-batch and inter-batch variation coefficients less than 15×, and the recovery rate between 80× and 120×. The matrix effect was further optimized to make it suitable for saliva sample detection. Saliva samples from individuals in different stress states were collected, and the CHGA levels were detected using the method established in this study, indicating its potential to reflect the intensity of stress. Conclusion A reliable saliva CHGA ELISA detection method has been successfully established, and its potential as a biomarker in stress-related research has been preliminarily explored.
Saliva/metabolism* ; Enzyme-Linked Immunosorbent Assay/methods* ; Humans ; Animals ; Mice, Inbred BALB C ; Mice ; Chromogranin A/immunology* ; Antibodies, Monoclonal/immunology* ; Female ; Male ; Reproducibility of Results ; Adult

The regulatory processes in developmental biology research are significantly influenced by long non-coding RNAs (lncRNAs). However, the dynamics of lncRNA expression during human tooth development remain poorly understood. In this research, we examined the lncRNAs present in the dental epithelium (DE) and dental mesenchyme (DM) at the late bud, cap, and early bell stages of human fetal tooth development through bulk RNA sequencing. Developmental regulators co-expressed with neighboring lncRNAs were significantly enriched in odontogenesis. Specific lncRNAs expressed in the DE and DM, such as PANCR, MIR205HG, DLX6-AS1, and DNM3OS, were identified through a combination of bulk RNA sequencing and single-cell analysis. Further subcluster analysis revealed lncRNAs specifically expressed in important regions of the tooth germ, such as the inner enamel epithelium and coronal dental papilla (CDP). Functionally, we demonstrated that CDP-specific DLX6-AS1 enhanced odontoblastic differentiation in human tooth germ mesenchymal cells and dental pulp stem cells. These findings suggest that lncRNAs could serve as valuable cell markers for tooth development and potential therapeutic targets for tooth regeneration.
Humans ; RNA, Long Noncoding/metabolism* ; Odontogenesis/genetics* ; Tooth Germ/embryology* ; Cell Differentiation ; Gene Expression Regulation, Developmental ; Mesoderm/metabolism* ; Tooth/embryology* ; Gene Expression Profiling ; Sequence Analysis, RNA ; Dental Pulp/cytology*

Objective:To discuss the regulatory role of complement alternative pathway in mouse colorectal cancer(CRC)liver metastasis model based on bioinformatics methods,and to clarify its mechanism through experimental verification.Methods:Using"CRC liver metastasis"as the keyword,the GSE81558 dataset was retrieved from Gene Expression Omnibus(GEO)database,including normal colon tissue samples,CRC tissue samples and CRC liver metastasis tissue samples.Bioinformatics methods were used to analyze and screen differentially expressed genes(DEGs).Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using R and Cytoscape software,and the results were visualized.Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)database was used to evaluate protein-protein interactions(PPIs)of DEGs and construct PPI network.Twelve C57BL/6 mice were injected with SL4 tumor cells into spleen,and the liver tissues were collected at 0,7 and 14 d.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of complement pathway-related genes in liver metastatic foci.The CRC liver metastasis mouse model was used to verify the complement signaling pathway.The mice were divided into control group,factor B knockout group(FB-/-)and C4 factor knockout group(C4-/-),and there were 6 mice in each group.The liver weights of the mice were measured;HE staining was used to detect the percentage of metastatic area in liver tissue in control group and FB-/-group;immunohistochemistry was used to detect macrophage infiltration in liver tissue in control group and FB-/-group,and the percentage of macrophage infiltration was calculated.Results:The distances between normal colon tissue samples and CRC tissue samples,as well as between CRC tissue samples and CRC liver metastasis tissue samples were far,indicating significant differences between samples,allowing subsequent analysis of DEGs.A total of 1 908 DEGs were screened in the dataset comparing normal colon tissue samples and CRC tissue samples,including 771 up-regulated DEGs and 1 137 down-regulated DEGs.Twenty-three up-regulated DEGs and 100 down-regulated DEGs were identified in the dataset comparing CRC and CRC liver metastasis.The GO functional enrichment analysis results showed that compared with normal colon tissue samples,DEGs in CRC samples were mainly enriched in biological processes(BP)related to cell cycle and mitosis,including mitotic cell cycle process,cell division,response to hormone,mitotic nuclear division and response to lipid.Compared with CRC samples,the DEGs in CRC liver metastasis samples were mainly enriched in coagulation-related BP,including platelet degranulation,blood coagulation regulation,acute-phase response,hemostasis regulation and coagulation regulation.The KEGG pathway enrichment analysis results showed that compared with normal colon tissue samples,the DEGs in CRC tissue samples were mainly enriched in cell cycle and p53 signaling pathways.Compared with CRC tissue samples,the DEGs in CRC liver metastasis tissue samples were mainly enriched in complement,coagulation cascade and metabolism-related signaling pathways.The Hub genes identified in PPI network were related to blood proteins.The RT-qPCR results showed that compared with 0 d group,the mRNA expression level of complement related genes complement 1q(C1q)in liver metastatic foci tissue sampres in 7 d group was significantly decreased(P<0.05),the mRNA expression levels of complement 3(C3),complement 5(C5),FB,and factor D(FD)were significantly increased(P<0.05 or P<0.01),the mRNA expression levels of complement pathway-related genes C1q,complement 2(C2),C3,complement fragment 3a receptor(C3aR),C5,complement fragment 5a receptor(C5aR),decay-accelerating factor(DAF),FB and FD in liver metastatic foci tissue sampres in 14 d group were significantly increased(P<0.05 or P<0.01).Compared with control group,the liver weight of the mice in FB-/-group was significantly decreased(P<0.01),while there was no significant difference was observed in C4-/-group(P>0.05).The HE staining results showed that compared with control group,the liver metastatic foci in FB-/-mice were significantly decreased,and the percentage of metastatic area was decreased(P<0.01).The immunohistochemistry results showed that compared with control group,the macrophage infiltration in liver metastatic foci of the mice in FB-/-group was reduced,and the percentage of macrophage infiltration was decreased(P<0.01).Conclusion:Complement cascade is associated with CRC liver metastasis,and the alternative complement pathway regulates CRC liver metastasis,suggesting this pathway may serve as a potential therapeutic target for CRC liver metastasis.

Objective: To determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability. Methods: A modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels. Results: Tu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment. Conclusion TF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

Objective To explore the application of sensory integration training and Beckman mouth muscle training in children with language delay.Methods A total of 120 children with language delay who were admitted to Nanjing Tongren Hospital and Southeast University Affiliated Zhongda Hospital from March 2018 to March 2023 were selected as research objects.They were divided into control group and observation group according to a random number table,with 60 cases in each group.The control group received Beckman mouth muscle training,and the observation group received sensory integration training and Beckman mouth muscle training.The Gesell Developmental Schedule(Gesell)scores,sign-significant relation(S-S)scores,and Beckman mouth motor assessment scale scores were compared between the two groups before and after training.Results The scores of major motor activity,individual social behavior,language,adaptive behavior,and gross and fine motor activity were increased after training in both groups,and these scores in the observation group were significantly higher than those in the control group(all P<0.05).The DQ values of language comprehension,language expression,and operational capacity were increased after training in both groups,and these values in the observation group were significantly higher than those in the control group(all P<0.05).The tongue,cheek and lip movement scores were increased after training in both groups,and these scores in the observation group were significantly higher than those in the control group(all P<0.05).Conclusion Sensory integration training and Beckman mouth muscle training can effectively improve language development and mouth motor function,and promote language comprehension,language expression,and operational capacity in children with language delay.

Objective To investigate the value of Baveno Ⅶ criteria versus Expanded Baveno Ⅶ criteria in screening for high-risk varices (HRV) in patients with compensated advanced chronic liver disease (cACLD). Methods A total of 146 patients with cACLD who were admitted to Beijing Shijitan Hospital, Capital Medical University, from January 2016 to December 2018 were enrolled, and according to the absence or presence of HRV based on gastroscopy, they were divided into HRV group with 68 patients and control group with 78 patients. Clinical data, liver stiffness measurement (LSM), and gastroscopy findings were analyzed, and different Baveno Ⅶ criteria were analyzed in terms of their sensitivity and specificity in the diagnosis of HRV. The Mann-Whitney U test and the McNemar test were used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data between groups. A univariate logistic regression analysis was performed for the variables used to predict HRV, and a multivariate analysis was performed for the variables with P < 0.1. The two sets of Baveno Ⅶ criteria were compared in terms of their sensitivity and specificity in the diagnosis of HRV. Results A total of 146 patients were enrolled in the study, among whom 68 (46.6%) were found to have HRV. The median age was 54 years (range 29-84 years), male patients accounted for 65.8%, and hepatitis B virus was the main etiology observed in 115 patients (78.8%). The univariate logistic regression analysis showed that LSM and platelet count (PLT) were associated with HRV (both P < 0.05). The multivariate analysis showed that based on Baveno Ⅶ criteria, LSM > 20 kPa or PLT < 150×10 9 /L was associated with HRV (both P < 0.05), and based on Expanded Baveno Ⅶ criteria, LSM > 25 kPa or PLT < 110×10 9 /L was associated with HRV (both P < 0.05). LSM and PLT had an area under the ROC curve of 0.797 (95% confidence interval [ CI ]: 0.723-0.859) and 0.789 (95% CI : 0.714-0.852), respectively, in the diagnosis of HRV. There were significant differences in the prevalence rates of esophageal and gastric varices and HRV between the patients who met Baveno Ⅶ criteria and those who did not meet such criteria ( χ 2 =23.14 and 23.14, both P < 0.001), as well as between the patients who met Expanded Baveno Ⅶ criteria and those who did not meet such criteria ( χ 2 =43.51 and 25.71, both P < 0.001). Although a higher proportion of patients were exempted from gastroscopy based on Expanded Baveno Ⅶ criteria (32.9% vs 13.7%), Baveno Ⅶ criteria had higher sensitivity (0.98 vs 0.88) and negative predictive value (0.95 vs 0.83) and could better avoid the missed diagnosis of HRV (1.0% vs 9.3%). Conclusion Baveno Ⅶ criteria are more suitable for the screening for HRV in cACLD patients in China.

Objective:To explore the needs of parents of hospitalized neonates with the challenges of implementing family-centered care during the Covid-19 pandemic.Methods:Using a method of phenomenological interviewing and Colaizzi′s method of data analysis, the information of 18 parents of admitted infants of Children′s Hospital of Fudan University from January 1 to 20, 2022 were collected and analyzed.Results:In the post-epidemic era, 5 themes of needs for parents of hospitalized neonates during family-centered care were identified: closeness to babies; emotional support; training about feeding; accommodation services; financial support.Conclusions:In the post-epidemic era, experiencing worry, anxiety, uncertainty, helplessness, loss and other negative psychological experience, the parents of hospitalized neonates have many unsatisfied needs. Hospital administrators need to focus on the needs of parents for family-centered nursing care, and actively explore effective coping strategies.

Objective:To investigate the incidences of metachronous advanced adenoma (MAA) in patients with simultaneous multiple primary colorectal cancer (CRC) and patients with sporadic CRC.Methods:From January 1, 2008 to September 30, 2022, at Beijing Shijitan Hospital, Capital Medical University, CRC patients who underwent surgery and 3 years follow-up with endoscopy were enrolled. The patients completed colonoscopy at least 2 times during follow-up in 6 to 36 months after surgery, and the interval between the 2 times colonoscopies was over 6 months. Clinical data including age, gender, and tumor location, stage, pathological features, combined underlying diseases, preoperative carcinoembryonic antigen, hemoglobin and other laboratory results, baseline colonoscopy results, and detection of MAA were collected. According to age (±2 years old), gender, location of primary lesion and stage of tumor, patients with simultaneous CRC or sporadic CRC were matched at 1∶1 ratio by propensity score matching. The cumulative risks of MAA in patients with simultaneous multiple primary CRC and patients with sporadic CRC were calculated. Cox proportional hazard regression was used to analyze the influencing factors in the occurrence of MAA.Results:A total of 814 CRC patients were enrolled and matched. After paired matching, there were 36 cases of simultaneous multiple primary CRC (78 lesions) and 78 cases of sporadic CRC (78 lesions). The cumulative incidences of MAA at 1, 2 and 3 years of simultaneous CRC group were 11.1%(4/36), 22.2%(8/36) and 33.3%(12/36), respectively. The cumulative incidences of MAA at 1-, 2- and 3-year of sporadic CRC group were 3.8%(3/78), 12.8%(10/78) and 20.5%(16/78), respectively.Simultaneous CRC was correlated with an increase in the 3-year cumulative incidence of MAA ( HR=4.163, 95% confidence interval(95% CI) 1.032 to 4.721, P=0.047). Especially in left-sided CRC, the risk of MAA in simultaneous CRC increased ( HR=7.186, 95% CI 1.602 to 20.787, P=0.010). The results of multivariate cox-regression analysis indicated that detection of simultaneous advanced adenoma at baseline endoscopy was an independent risk factor of MAA ( HR=3.175, 95% CI 1.411 to 7.142, P=0.005). Conclusion:Colouoscopy follow-up should be strengthened in patients with simultaneous multiple primary CRC and simultaneous advanced adenomas.

Objective : To explore the possible mechanism of glycyrrhetinic acid on inhibiting malignant biological behaviors of melanoma by Wnt / β-catenin pathways． Methods: The melanoma cells B16-F10 were selected as the research objects．The concentration gradient tests (0，1，2，4 μmol /L) were conducted by MTT．The cells given cisplatin intervention was enrolled as positive controls．The cells invasion and migration were detected by Transwell chamber assay．The expression levels of Wnt / β-catenin pathway proteins，invasion and migration related proteins (MMP-2，MMP-9) in B16-F10 cells were detected by Wester blot．The xenograft models of nude mice were constructed，and they were divided into control group (without drugs treatment) and glycyrrhetinic acid group (40 mg / kg) ．The growth of tumor tissues，and expression levels of Wnt / β-catenin pathway proteins，invasion and migration related proteins were observed. Results : MTT results showed that glycyrrhetic acid could inhibit the proliferation of B16-F10 cells in a concentration-dependent manner．The inhibition effect of glycyrrhetic acid ( ≥2 μmol /L) was significant on the proliferation of B16-F10 cells (P ＜0. 05) ．The results of Transwell chamber assay showed that compared with control group，invasion and migration abilities of B16-F10 cells were significantly reduced after treatment with glycyrrhetinic acid (2，4 μmol /L) (P ＜0. 05) ．Wester blot results showed that compared with those without glycyrrhetinic acid treatment，expression levels of MMP-9 ，MMP-2，Wnt1 and β-catenin protein in B16- F10 cells significantly decreased after treatment with glycyrrhetinic acid (2，4 μmol /L) (P＜0. 05) ．The results of tumor-bearing assay showed that compared with control group ，weight and volume of tumors significantly decreased in glycyrrhetinic acid group，and expression levels of Wnt1 ，β-catenin，MMP-9 and MMP-2 proteins also significantly decreased (P＜0. 05) ． Conclusion Glycyrrhetinic acid can significantly inhibit the malignant biological behaviors of melanoma in vitro and vivo．And its mechanism may be related to inhibiting the activation of Wnt / β-catenin signaling pathways.