[Objective] To resolve the ambiguities of HLA genotyping generated by next generation sequencing (NGS) using nanopore sequencing technology. [Methods] A total of 38 samples with ambiguous HLA genotyping by NGS in our laboratory were collected, and HLA-A, -B, -C, -DRB1, -DRB3/4/5, -DQA1, -DQB1, -DPA1 and -DPB1 loci in these samples were amplified using primers in the same commercial NGS HLA genotyping kit, then subjected to third-generation library construction, and sequenced on the nanopore sequencer. The sequencing data were converted into Fastq files and analyzed by software, and the genotypes of 11 HLA loci were obtained. The ambiguities were counted directly. [Results] The high-resolution genotyping at the second domain of 11 HLA loci of 38 samples using the third generation sequencing (TGS) were consistent with the results of the NGS method at a rate of 100%. The genotypes for the HLA-A, -B, -C, -DRB3, -DRB4, -DQA1 and -DPA1 loci by TGS were all only one result, and the discrimination rate for ambiguities of the HLA-A, -B, -C, and -DQA1 loci (all caused by the difficulty in phasing due to the short NGS read length) was 100%. Among the HLA-DRB1, -DRB5, -DQB1 and -DPB1 loci, the discrimination rate of TGS for the ambiguities caused by non-amplification of exon 1 was 0% and by the short NGS read length was 100%. [Conclusion] Nanopore technology was used to identify the ambiguities of 11 HLA loci in this study, and the ambiguities caused by the short read length disadvantage of the NGS method could be solved effectively and the accuracy of HLA genotyping would be improved.

Cardiac fibrosis is characterized by an elevated amount of extracellular matrix (ECM) within the heart. However, the persistence of cardiac fibrosis ultimately diminishes contractility and precipitates cardiac dysfunction. Circular RNAs (circRNAs) are emerging as important regulators of cardiac fibrosis. Here, we elucidate the functional role of a specific circular RNA CELF1 in cardiac fibrosis and delineate a novel feedback loop mechanism. Functionally, circ-CELF1 was involved in enhancing fibrosis-related markers' expression and promoting the proliferation of cardiac fibroblasts (CFs), thereby exacerbating cardiac fibrosis. Mechanistically, circ-CELF1 reduced the ubiquitination-degradation rate of BRPF3, leading to an elevation of BRPF3 protein levels. Additionally, BRPF3 acted as a modular scaffold for the recruitment of histone acetyltransferase KAT7 to facilitate the induction of H3K14 acetylation within the promoters of the Celf1 gene. Thus, the transcription of Celf1 was dramatically activated, thereby inhibiting the subsequent response of their downstream target gene Smad7 expression to promote cardiac fibrosis. Moreover, Celf1 further promoted Celf1 pre-mRNA transcription and back-splicing, thereby establishing a feedback loop for circ-CELF1 production. Consequently, a novel feedback loop involving CELF1/circ-CELF1/BRPF3/KAT7 was established, suggesting that circ-CELF1 may serve as a potential novel therapeutic target for cardiac fibrosis.

Digital technologies have become an integral part of complete denture restoration. With advancement in computer-aided design and computer-aided manufacturing (CAD/CAM), tools such as intraoral scanning, facial scanning, 3D printing, and numerical control machining are reshaping the workflow of complete denture restoration. Unlike conventional methods that rely heavily on clinical experience and manual techniques, digital technologies offer greater precision, predictability, and efficacy. They also streamline the process by reducing the number of patient visits and improving overall comfort. Despite these improvements, the clinical application of digital complete denture restoration still faces challenges that require further standardization. The major issues include appropriate case selection, establishing consistent digital workflows, and evaluating long-term outcomes. To address these challenges and provide clinical guidance for practitioners, this expert consensus outlines the principles, advantages, and limitations of digital complete denture technology. The aim of this review was to offer practical recommendations on indications, clinical procedures and precautions, evaluation metrics, and outcome assessment to support digital restoration of complete denture in clinical practice.
Humans ; Denture, Complete ; Computer-Aided Design ; Denture Design/methods* ; Consensus ; Printing, Three-Dimensional

Objective:To investigate the clinicopathological features and differential diagnosis of primary mucosal CD30-positive T-cell lymphoproliferative disorders (pmCD30 +TLPD). Methods:Eight cases of pmCD30 +TLPD diagnosed from 2013 to 2023 at the Department of Pathology, Beijing Friendship Hospital Affiliated to Capital Medical University and Beijing Ludaopei Hospital were retrospectively collected. The immunophenotype, EBV infection status and T-cell receptor (TCR) clonability of tumor cells were examined. The clinicopathological features were analyzed and related literatures were reviewed. Results:There were 5 females and 3 males, aged 28 to 73 years, without B symptoms, lack of trauma and autoimmune diseases. Seven cases occurred in oral mucosa and one in anal canal mucosa. Submucosal nodules with ulcerations were presented in all cases except one, which only submucosal nodule. Morphologically, there was different distribution of allotypic lymphocytes in inflammatory background. Four cases showed “kidney-shaped”, “embryonic” and “horseshoe-shaped” cells, and one case resembled Hodgkin and Reed/Sternberg (HRS) cells. Allotypic lymphocytes expressed CD3 (7/8), CD4+/CD8-(7/8) and CD4-/CD8-(1/8). CD30 was uniformly strongly positive while ALK and CD56 were negative. In situ hybridization of EBER was negative in five cases (5/5). Clonal TCR gene rearrangement was positive in two cases. Four patients did not receive radiotherapy or chemotherapy. All the seven patients survived without disease except one died due to concurrent leukopenia.Conclusions:pmCD30 +TLPD had a broad morphological spectrum and could be easily confused with primary cutaneous CD30 +TLPD and systemic ALK-negative anaplastic large cell lymphoma involving mucosa, which may lead to misdiagnosis. Although the majority of the cases had a favorable prognosis, a few cases relapsed or progressed to lymphoma.

【Objective】 To understand factors associated with children′s attendance at nursery care institutions (NCIs) and the basic characteristics of the NCIs children were enrolled in, so as to provide scientific evidence for policymakers. 【Methods】 During November 2020 and January 2021, parents who visited the Department of Child Health Care in six selected maternal and children′s hospitals, as well as nearby NCIs in Zhejiang were invited to finish an online questionnaire. Information such as children′s sociodemographic characteristics, parents′ knowledge, attitude and practice regarding nursery and feeding, etc. were collected. A total of 1 756 questionnaires were collected. 【Results】 Compared to children who were not in NCIs, children enrolled in NCIs were older (94.4% of children ≥24 months vs. 30.1%, χ²=835.27), more likely to be from the local area (87.2% vs. 81.4%,χ²=12.25), more likely to have parents with a college degree (mother: 83.6% vs. 74.2%, χ²=35.29; father: 79.9% vs. 70.0%, χ²=27.01), had a higher prevalence of family annual income >200 000 CNY (49.5% vs. 28.2%, χ²=110.49), and were less likely to have their grandparents available to take care of them (16.7% vs. 26.8%, χ²=31.4) The difference all have great significant.(P<0.05). In a multivariate Logistic regression model, the older the child, the more likely they were to attend an NCI (for children aged 6 - 23 months, OR=6.70; for children aged 24 - 35 months, OR=134.03; and for children aged 36 - 42 months, OR=699.33; P<0.05). Family annual income was positively associated with children′s attendance at NCIs (for those earning 100 000 - 200 000 CNY/year, OR=1.63; for those earning 200 000 - 500 000 CNY/year, OR=2.96; and for those earning >500 000 CNY/year, OR=4.62, P<0.05). Conversely, the higher the level of grandparent involvement in daily care, the lower the attendance at NCIs (for children cared for by both parents and grandparents, OR=0.57; for those primarily cared for by grandparents, OR=0.26, P<0.05). For children who used to stay at NCIs, 82.8% stayed at institutions that only recruited children aged 0 - 3 years, 97.4% spent their whole day in NCIs, and 71.4% spent less than 3 000 CNY per month for NCI services. Additionally, over 95% of parents were satisfied with the food and care services in NCIs, as well as their children′s physical development in NCIs. However, 32.1% of NCIs were reported by parents as having no room for breastfeeding. 【Conclusions】 Children′s age, grandparent involvement in routine care, and family annual income are the main factors associated with children′s attendance at NCIs. There is a greater need for more affordable and community-based NCIs, particularly for children under 2 years old. Additionally, more attention should be paid to the quality surveillance, assessment and management of NCIs.

OBJECTIVE To investigate the effects of erianin （ERI） on the apoptosis of ovarian granulosa cells in rats with polycystic ovary syndrome （PCOS） and its mechanism. METHODS PCOS rat model was constructed by subcutaneous injection of dehydroepiandrosterone， and the successfully constructed rats were randomly divided into PCOS group， ERI low-dose， medium- dose and high-dose groups （10， 20， 40 mg/kg） and ERI high dose + verteporfin group （40 mg/kg ERI + 10 mg/kg verteporfin）， with 10 rats in each group. Another 10 normal rats were selected as the normal group. Rats in each administration group were given corresponding dose of ERI and/or intraperitoneal injection of vitiporfin， and rats in the PCOS group and normal group were orally administered an equal volume of 1% dimethyl sulfoxide， once a day， for 6 consecutive weeks. After administration， the body weight， fasting blood glucose （FPG）， serum levels of estradiol （E2）， testosterone （T）， follicle stimulating hormone （FSH） and luteinizing hormone （LH） were detected in each group； morphological changes in ovarian tissue were observed， and the apoptosis of ovarian tissue cells was analyzed. Apoptosis-associated proteins [B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， Caspase-3] and Hippo-YAP signaling pathway associated proteins [large tumor suppressor kinase 1 （LATS1）， phosphorylated LATS1 （p-LATS1） and Yes associated protein （YAP）， phosphorylated YAP （p-YAP）， transcriptional co-activator with PDZ binding motif （TAZ）] were detected in ovarian tissue. RESULTS Compared with PCOS group， the ovarian polycystic characteristics of the ERI low-dose， medium-dose，and high-dose groups were reduced， the number of atretic follicles was reduced， and the granulosa cell layer was thickened； the body mass， FPG， T， LH， LH/FSH， the number of cystic follicles， cell apoptosis index， protein expressions of Bax， Caspase-3， p-LATS1 and p-YAP were greatly decreased （P＜0.05）； the number of corpus luteum， protein expressions of E2， Bcl-2， LATS1， YAP and TAZ were greatly increased （P＜0.05）. Compared with ERI high-dose group， the above indexes in ERI high-dose + vitiporfin group were inhibited （P＜0.05）. CONCLUSIONS ERI can promote the proliferation of ovarian granulosa cells and improve the level of sex hormones in PCOS rats， and its mechanism of action may be related to the inhibition of the Hippo-YAP signaling pathway.

OBJECTIVE To investigate the effects of erianin （ERI） on the apoptosis of ovarian granulosa cells in rats with polycystic ovary syndrome （PCOS） and its mechanism. METHODS PCOS rat model was constructed by subcutaneous injection of dehydroepiandrosterone， and the successfully constructed rats were randomly divided into PCOS group， ERI low-dose， medium- dose and high-dose groups （10， 20， 40 mg/kg） and ERI high dose + verteporfin group （40 mg/kg ERI + 10 mg/kg verteporfin）， with 10 rats in each group. Another 10 normal rats were selected as the normal group. Rats in each administration group were given corresponding dose of ERI and/or intraperitoneal injection of vitiporfin， and rats in the PCOS group and normal group were orally administered an equal volume of 1% dimethyl sulfoxide， once a day， for 6 consecutive weeks. After administration， the body weight， fasting blood glucose （FPG）， serum levels of estradiol （E2）， testosterone （T）， follicle stimulating hormone （FSH） and luteinizing hormone （LH） were detected in each group； morphological changes in ovarian tissue were observed， and the apoptosis of ovarian tissue cells was analyzed. Apoptosis-associated proteins [B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， Caspase-3] and Hippo-YAP signaling pathway associated proteins [large tumor suppressor kinase 1 （LATS1）， phosphorylated LATS1 （p-LATS1） and Yes associated protein （YAP）， phosphorylated YAP （p-YAP）， transcriptional co-activator with PDZ binding motif （TAZ）] were detected in ovarian tissue. RESULTS Compared with PCOS group， the ovarian polycystic characteristics of the ERI low-dose， medium-dose，and high-dose groups were reduced， the number of atretic follicles was reduced， and the granulosa cell layer was thickened； the body mass， FPG， T， LH， LH/FSH， the number of cystic follicles， cell apoptosis index， protein expressions of Bax， Caspase-3， p-LATS1 and p-YAP were greatly decreased （P＜0.05）； the number of corpus luteum， protein expressions of E2， Bcl-2， LATS1， YAP and TAZ were greatly increased （P＜0.05）. Compared with ERI high-dose group， the above indexes in ERI high-dose + vitiporfin group were inhibited （P＜0.05）. CONCLUSIONS ERI can promote the proliferation of ovarian granulosa cells and improve the level of sex hormones in PCOS rats， and its mechanism of action may be related to the inhibition of the Hippo-YAP signaling pathway.

Objective:To explore the anxiety level, influencing factors among surrogate decision-makers of patients with acute ischemic stroke during thrombolysis decision-making, and their correlation with decision-making duration.Methods:Acute ischemic stroke patients and their surrogate decision-makers who visited the Emergency Department of the First Affiliated Hospital of Zhengzhou University from September 2019 to December 2021 were selected as the research subjects.Sociodemographic data and disease related data of patients and surrogate decision-makers were collected.Surrogate decision-makers were evaluated with the state-trait anxiety inventory, decision participation expectation scale, Wake Forest physician trust scale, and perceived social support scale.SPSS 26.0 software was used for data processing.Pearson correlation analysis, Spearman correlation analysis and ridge regression analysis were used for statistical analysis.Results:The score of state anxiety of decision-makers was (49.47±9.04), and 18.2% (70/383) of decision-makers had a decision duration exceeding 15 minutes.The score of state anxiety of decision-makers was positively correlated with decision duration ( r=0.189, P<0.001). The influencing factors of state anxiety level of decision-makers included sociodemographic factors (age of decision-makers and patients, relationship between payers and patients, whether decision-makers bear the current medical expenses, type of medical insurance for patients), psychological factors (trust level in physicians, perceived social support), factors related to patient disease (numbers of stroke relapses, National Institutes of Health stroke scale scores for patients), characteristics of the decision-making process (whether patients participate in the decision-making process, and the role of decision-makers in the decision-making process) (all P<0.05). Conclusion:Most surrogate decision-makers experience anxiety.Medical staff should pay attention to the emotions of decision-makers and adopt appropriate communication skills when communicating with informed consent for thrombolysis, alleviate the anxiety of surrogate decision-makers, so as so reduce the decision-making duration.

The cognitive impairment of diabetes mellitus type 2(T2DM)is closely related to neurovascular coupling(NVC)changes,but the exact mechanism remains unclear.Functional MRI(fMRI)technology were able to jointly analyze NVC changes of T2DM,providing new ideas for revealing the mechanism of cognitive dysfunction caused by T2DM.The progresses of fMRI for exploring NVC changes in T2DM were reviewed in this article.

Objective:To analyze the current status of internet-based patient education materials related to labor analgesia in China.Methods:Labor analgesia-related materials were retrieved and screened according to the search habits of Chinese search engine users. The coverage and accuracy of the content were evaluated by 3 anesthesiologists. The Chinese version of the Patient Education Materials Assessment Tool for Print Materials was used to subjectively assess comprehensibility and operability from a medical text perspective. The consistency of the evaluation results of the three anesthesiologists was analyzed using the intraclass correlation coefficient method. A machine learning method combined with ChatGPT-4.0 was employed to establish a Chinese readability classification model to objectively evaluate the readability difficulty of the included materials from a Chinese text perspective.Results:A total of 97 web pages were retrieved, with 21 valid materials included in the study. The coverage rate of contraindications for labor analgesia was only 62% (13/21), and the accuracy rate of materials exceeding 90% was 71% (15/21). Internet-based materials that were easy to understand accounted for 81% (17/21), while the constituent ratio of internet-based materials with instructional significance were only 5% (1/21). The intraclass correlation coefficient values of consistency evaluation for coverage rate, accuracy rate, and comprehensibility and operability were 0.975, 0.833, 0.758, and 0.773, respectively ( P<0.001). Internet-based materials suitable for compulsory education level were only 5% (1/21), while those suitable for high school and above education level accounted for 43% (9/21). Conclusions:There are numerous internet-based patient education materials related to labor analgesia in China, but the quality needs improvement. In the future, a collaborative model of " anesthesiology+ linguistics" should be developed to provide patients with more comprehensive, accurate, and pregnant-friendly patient education materials.