Aim To investigate the effect of N6-methy-ladenosine(m6A)demethylase ALKBH5 on the prolif-eration and migration of cardiac fibroblasts(CFs)in-duced by high glucose.Methods Primary CFs were isolated from neonatal mouse hearts and identified u-sing optical and confocal microscopy.Cell activation was induced using a high-glucose medium(33 mmol·L-1 glucose).An ALKBH5 overexpression model was established by transfecting CFs with an ALKBH5 ex-pression vector in a high-glucose medium.The expres-sion of ALKBH5 in CFs was assessed through immuno-fluorescence staining,Western blot and RT-qPCR.Changes in m6A levels were evaluated using Dot blot a-nalysis.Additionally,Alterations in the expression of proliferating cell nuclear antigen(PCNA)and collagenⅠ,a pivotal fibrosis indicator,were measured using Western blot.The proliferation and migration ability of CFs were assessed through EdU staining and Transwell migration assay,respectively.Results Following treatment with high glucose,the expression of ALKBH5 in CFs notably decreased,while m6A level increased.This was accompanied by a significant increase in the expression of the proliferation marker PCNA and the fi-brosis marker collagen Ⅰ.Additionally,there was a sig-nificant improvement in the ability of proliferation and migration.Overexpression of ALKBH5 resulted in a significant decrease in the expressions of PCNA and collagen Ⅰ,leading to the inhibition of both proliferation and migration in CFs.Conclusion Overexpression of ALKBH5 suppresses the expression of PCNA and colla-gen Ⅰ,consequently reducing the proliferation and mi-gration of CFs,potentially through m6A methylation modification.

Aim To investigate the effect of N6-methy-ladenosine(m6A)demethylase ALKBH5 on the prolif-eration and migration of cardiac fibroblasts(CFs)in-duced by high glucose.Methods Primary CFs were isolated from neonatal mouse hearts and identified u-sing optical and confocal microscopy.Cell activation was induced using a high-glucose medium(33 mmol·L-1 glucose).An ALKBH5 overexpression model was established by transfecting CFs with an ALKBH5 ex-pression vector in a high-glucose medium.The expres-sion of ALKBH5 in CFs was assessed through immuno-fluorescence staining,Western blot and RT-qPCR.Changes in m6A levels were evaluated using Dot blot a-nalysis.Additionally,Alterations in the expression of proliferating cell nuclear antigen(PCNA)and collagenⅠ,a pivotal fibrosis indicator,were measured using Western blot.The proliferation and migration ability of CFs were assessed through EdU staining and Transwell migration assay,respectively.Results Following treatment with high glucose,the expression of ALKBH5 in CFs notably decreased,while m6A level increased.This was accompanied by a significant increase in the expression of the proliferation marker PCNA and the fi-brosis marker collagen Ⅰ.Additionally,there was a sig-nificant improvement in the ability of proliferation and migration.Overexpression of ALKBH5 resulted in a significant decrease in the expressions of PCNA and collagen Ⅰ,leading to the inhibition of both proliferation and migration in CFs.Conclusion Overexpression of ALKBH5 suppresses the expression of PCNA and colla-gen Ⅰ,consequently reducing the proliferation and mi-gration of CFs,potentially through m6A methylation modification.

Blau syndrome is a rare genetic disorder characterized by the a mix of granulomatous arthritis, uveitis, and dermatitis. Patients typically manifest multisystem involvement, including ocular, skin, and skeletal abnormalities. Blau syndrome is extremely rare, with a global incidence of less than one in a million among children. In this multidisciplinary consultation, we present a case of a 21-year-old young female patient having multisystemic involvement since early childhood. She was presented with multiple joint swelling, skin lesions, increased eye discharge, and accompanied by hypertension and arterial abnormalities, and received a diagnosis of uveitis. The patient had been receiving steroid treatment since the age of 6 and has tried various medications, with some improvement in joint swelling and ocular symptoms. Through this rare disease multidisciplinary consultation, we aim to provide guidance in the molecular diagnosis of the patient, multisystem assessment, and the selection and formulation of treatment plans. Additionally, we hope that by reporting this case, clinical physicians can gain a better understanding of the diagnosis and comprehensive treatment strategies for Blau syndrome, thereby improving the management and treatment of rare diseases.

Objective: To evaluate the application effect of smart classroom teaching mode in undergraduate teaching of endodontics. Methods: Through micro-lecture and massive open online course which were closely integrated with clinical practice and frontier advances, we build a new smart classroom teaching mode of endodontics relying on information technology such as the medical education cloud APP platform. The mode was applied to the undergraduate teaching of grade 2017 (110 students) and grade 2018 (107 students) in 2020 and 2021 respectively (experimental group). The theoretical examination was conducted for the grade 2016 (control group, 111 students applied traditional teaching methods) in 2019, and for two experimental grades in 2020 and 2021 respectively. A questionnaire survey was conducted for the 2018 undergraduates to investigate the experience of the smart classroom teaching mode, and the application effect of the smart classroom teaching mode was evaluated by comparing the offline theoretical test scores of grades 2016, 2017 and 2018. Results: The results of the questionnaire showed that students in grade 2018 recognized the overall form of smart classroom teaching mode, and 75.2% (79/105) of the students satisfied with the teaching process, considering that it could enhance learning interest and enthusiasm, improve self-learning ability, facilitate the understanding and memory of knowledge points, as well as increase the extension and expansion of professional knowledge. Thirty-seven point one percent (39/105) of the students thought that smart classroom teaching mode was not conducive to the interaction between teachers and students and couldn't improve learning efficiency. Comparing the final theoretical examination scores of students in three years, it was found that the average scores of 2021 (78.79±9.88) and 2020 (76.45±8.33) were significantly higher than that of 2019 (67.67±10.58) (t=6.77, P<0.001; t=8.51, P<0.001). The average score in 2021 was higher than that in 2020, although the difference was not significant (t=1.79, P=0.223). Conclusions: The application of smart classroom mode improved the teaching effect of endodontics, which is worthy of further promotion to provide a positive reference in improving the educating effects of oral medicine.
Humans ; Learning ; Endodontics ; Students ; Dental Care ; Surveys and Questionnaires

Tryptophan 2,3-dioxygnease 2 (TDO2) is specific for metabolizing tryptophan to kynurenine (KYN), which plays a critical role in mediating immune escape of cancer. Although accumulating evidence demonstrates that TDO2 overexpression is implicated in the development and progression of multiple cancers, its tumor-promoting role in esophageal squamous cell carcinoma (ESCC) remains unclear. Here, we observed that TDO2 was overexpressed in ESCC tissues and correlated significantly with lymph node metastasis, advanced clinical stage, and unfavorable prognosis. Functional experiments showed that TDO2 promoted tumor cell proliferation, migration, and colony formation, which could be prevented by inhibition of TDO2 and aryl hydrocarbon receptor (AHR). Further experimentation demonstrated that TDO2 could promote the tumor growth of KYSE150 tumor-bearing model, tumor burden of C57BL/6 mice with ESCC induced by 4-NQO, enhance the expression of phosphorylated AKT, with subsequent phosphorylation of GSK3

ObjectiveTo investigate the expression of diabetes related microRNA （miRNA） in plasma exosomes of pregnant women complicated with gestational diabetes mellitus （GDM） and to study their association with the pathogenesis of GDM. MethodsIn this study， diabetes related miRNAs microarray data sets were searched through Gene Expression Omnibus （GEO） database， and screened by GEO2R to determine the candidate miRNAs. Differentially expressed miRNAs were selected if the change is consistent in more than one microarray data sets. Singleton pregnant women within 10-16 gestational weeks （gws） were included and their plasma was obtained for further analysis. In 24~28 gws， oral glucose tolerance tests （OGTT） were performed to diagnose GDM or normal glucose tolerance （NGT）. Diabetes related miRNAs expression in plasmatic exosomes was compared between GDM and NGT groups by RT-PCR. Receiver operating characteristic （ROC） curve was applied to assess the predictive efficiency of miRNAs. Finally， Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis was used to reveal the function of miRNAs. ResultsFive diabetes related microarray datasets were downloaded from GEO database， including GSE94649， GSE21321， GSE98043， GSE148961 and GSE27645. Ten differentially expressed miRNAs were found， among which seven were up-regulated and three were down-regulated. RT-PCR results showed that， compared with NGT group， hsa-miR-188-5p， hsa-miR-663a， hsa-miR-135a-5p and hsa-miR-4707-5p in plasma exosomes of GDM pregnant women were up-regulated. The area under the ROC curve （AUC） of these four miRNAs for GDM prediction was 0.839 （95%CI：0.724~0.954）. The target genes of miR-135a-5p were involved in insulin signaling pathways. ConclusionsDifferentially expressed exosomal miRNAs in GDM plasma act as potential predictors of GDM. Among them， miR-135a-5p mainly participates in insulin signaling pathways.

ObjectiveTo investigate the expression of Ubiquitin specific protease 44（USP44）and its clinical significance in hepatocellular carcinoma（HCC）. MethodsImmunohistochemistry was used to detect the expression of USP44 protein in 161 cases of hepatocellular carcinoma. The correlation between the expression of USP44 protein and clinicopathological parameters，and its prognosis value were analyzed. ResultsImmunohistochemical analysis showed USP44 displayed elevated expression in 98 HCC cases out of 161 cases. Pearson′s chi-square test indicated the expression of USP44 in HCC was significantly correlated with clinical stage（χ²=14.44，P＜0.001） and tumor multiplicity（Unifocal or Multifocal）（χ² =8.04，P=0.005）. Kaplan-Meier analysis indicated patients with low USP44 expression correlated with poor overall survival（Log-rank χ²=20.77，P＜0.001）.The overall mean survival time was 59.6 months in low USP44 expression patients and 185.0 months in high USP44 expression patients. The Cox proportional hazard model analysis showed USP44 was an independent prognostic risk factor for HCC patients. ConclusionUSP44 is an independent prognostic risk factor for HCC patients， and the expression of USP44 protein could be used to screen patients with good prognosis for HCC. Our study may provide evidence for USP44 as a potential therapeutic target for HCC patients.

PD-1 and CTLA-4 antibodies offer great hope for cancer immunotherapy. However, many patients are incapable of responding to PD-1 and CTLA-4 blockade and show low response rates due to insufficient immune activation. The combination of checkpoint blockers has been proposed to increase the response rates. Besides, antibody drugs have disadvantages such as inclined to cause immune-related adverse events and infiltration problems. In this study, we developed a cyclic peptide C25 by using Ph.D.-C7C phage display technology targeting LAG-3. As a result, C25 showed a relative high affinity with human LAG-3 protein and could effectively interfere the binding between LAG-3 and HLA-DR (MHC-II). Additionally, C25 could significantly stimulate CD8 T cell activation in human PBMCs. The results also demonstrated that C25 could inhibit tumor growth of CT26, B16 and B16-OVA bearing mice, and the infiltration of CD8 T cells was significantly increased while FOXP3 Tregs significantly decreased in the tumor site. Furthermore, the secretion of IFN- by CD8 T cells in spleen, draining lymph nodes and especially in the tumors was promoted. Simultaneously, we exploited T cells depletion models to study the anti-tumor mechanisms for C25 peptide, and the results combined with MTT assay confirmed that C25 exerted anti-tumor effects CD8 T cells but not direct killing. In conclusion, cyclic peptide C25 provides a rationale for targeting the immune checkpoint, by blockade of LAG-3/HLA-DR interaction in order to enhance anti-tumor immunity, and C25 may provide an alternative for cancer immunotherapy besides antibody drugs.

Measurement of corpse temperature is mainly used for estimation of early postmortem interval, and rectal temperature is often used as a representative of body's core temperature in actual work because it is simple, quick and non-invasive. At present, the rectal temperature postmortem interval estimation method internationally accepted and widely used is HENSSGE's nomogram method, while many domestic scholars also deduced their own regression equations through a large number of case data. Estimation of postmortem interval based on rectal temperature still needs further study. The nomogram method needs to be optimized and extended, and quantification of its influencing factors needs to be dealt with more scientifically. There is still a lack of consensus on the probability and duration of the temperature plateau. There is no clear understanding of the probability and extent of the change in initial temperature caused by various causes. New methods and ideas enrich methodological research, but it still lacks systemicity and practicality. This article reviews the researches on estimation of postmortem interval based on rectal temperature in order to summarize the current situation of previous researches and seek new breakthrough points. Because the decline of body temperature can be easily influenced by many factors in vitro and vivo, and the influencing factors in different regions vary greatly, regionalization research and application may be a practical exploration to improve the accuracy of postmortem interval determination.
Autopsy ; Body Temperature ; Cadaver ; Humans ; Postmortem Changes ; Probability ; Temperature ; Time Factors

PURPOSE: Hepatocellular carcinoma (HCC) is an aggressive disease with high recurrence rate. However, current staging systems were lack of predictive capacity for HCC recurrence. We aimed to develop prognostic nomograms based on inflammation-related markers for HCC patients underwent hepatectomy. MATERIALS AND METHODS: We recruited 889 surgically treated patients from two medical centers. Independent prognostic factors were identified by cox regression analyses. Nomograms for recurrence-free survival (RFS) and overall survival (OS) were established, and validated internally and externally. The performance, discrimination, and calibration of nomograms were assessed, and compared with existed staging systems. RESULTS: Neutrophil to lymphocyte ratio (NLR) and gamma-glutamyl transpeptidase to platelet ratio (GPR) were the two inflammation-related factor that independently correlated with survival. NLR, GPR, international normalized ratio (INR), microvascular invasion, satellite lesions, tumour number, tumour diameter, and macrovascular invasion were used to construct nomogram for RFS while GPR, total bilirubin, INR, α-fetoprotein, microvascular invasion, satellite lesions, tumour diameter, and macrovascular invasion were for OS. In the training cohort, the C-index of nomogram was 0.701 (95% confidence interval [CI], 0.669 to 0.732) for RFS and 0.761 (95% CI, 0.728 to 0.795) for OS. These results received both internal and external validation with C-index of 0.701 (95% CI, 0.647 to 0.755) and 0.707 (95% CI, 0.657 to 0.756) for RFS, and 0.706 (95% CI, 0.640 to 0.772) and 0.708 (95% CI, 0.646 to 0.771) for OS, respectively. The nomograms showed superior accuracy to conventional staging systems (p<0.001). CONCLUSION: The nomograms based on inflammation-related markers are of high efficacy in predicting survival of HCC patients after hepatectomy, which will be valuable in guiding postoperative interventions and follow-ups.
Bilirubin ; Blood Platelets ; Calibration ; Carcinoma, Hepatocellular ; Cohort Studies ; Discrimination (Psychology) ; Follow-Up Studies ; gamma-Glutamyltransferase ; Hepatectomy ; Humans ; Inflammation ; International Normalized Ratio ; Lymphocytes ; Neutrophils ; Nomograms ; Recurrence