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*

Identification of disease-specific cell subtypes (DSCSs) has profound implications for understanding disease mechanisms, preoperative diagnosis, and precision therapy. However, achieving unified annotation of DSCSs in heterogeneous single-cell datasets remains a challenge. In this study, we developed the gPRINT algorithm (generalized approach for cell subtype identification with single cell's voicePRINT). Inspired by the principles of speech recognition in noisy environments, gPRINT transforms gene position and gene expression information into voiceprints based on ordered and clustered gene expression phenomena, obtaining unique "gene print" patterns for each cell. Then, we integrated neural networks to mitigate the impact of background noise on cell identity label mapping. We demonstrated the reproducibility of gPRINT across different donors, single-cell sequencing platforms, and disease subtypes, and its utility for automatic cell subtype annotation across datasets. Moreover, gPRINT achieved higher annotation accuracy of 98.37% when externally validated based on the same tissue, surpassing other algorithms. Furthermore, this approach has been applied to fibrosis-associated diseases in multiple tissues throughout the body, as well as to the annotation of fibroblast subtypes in a single tissue, tendon, where fibrosis is prevalent. We successfully achieved automatic prediction of tendinopathy-specific cell subtypes, key targets, and related drugs. In summary, gPRINT provides an automated and unified approach for identifying DSCSs across datasets, facilitating the elucidation of specific cell subtypes under different disease states and providing a powerful tool for exploring therapeutic targets in diseases.
Humans ; Algorithms ; Single-Cell Analysis ; Databases, Genetic ; Molecular Sequence Annotation

Objective To observe the value of intravascular ultrasound(IVUS)for assisting endovascular therapy for renal artery stenosis(RAS).Methods Thirty patients with RAS who underwent endovascular therapy were retrospectively analyzed.Parameters of renal artery and plaques in RAS segment measured with CT angiography(CTA)and IVUS before treatment were compared.Bland-Altman diagram was performed to evaluate the consistency of lumen cross-sectional stenosis rate and plaque eccentricity index between CTA and IVUS.The stent parameters measured with IVUS were recorded immediately after implantation of balloon-expandable covered stents.Results Before treatment,the minimum lumen diameter,lumen cross-sectional stenosis rate and stenotic segment length of IVUS were all larger,while maximum lumen diameter and lumen eccentricity index of IVUS were both smaller than those of CTA(all P＜0.05).No significant difference of plaque eccentricity index,plaque type nor stenosis distal remodeling was found between CTA and IVUS(all P＞0.05).The average difference between IVUS and CTA for evaluating lumen cross-sectional stenosis rate and plaque eccentricity index was-0.020(-0.096,0.050)and-0.020(-0.130,0.091),respectively.The consistency of IVUS and CTA for evaluating plaque eccentricity index was better than that of lumen cross-sectional stenosis rate.The stent symmetry,stent eccentricity index,stent expansion coefficient and stenosis coverage rate immediately after implantation measured with IVUS was(82.69±14.61)%,(1.54±9.16)%,(99.81±10.70)%and 100%,respectively.Among 30 cases,2 cases(2/30,6.67%)underwent postdilation since poor stent apposition.Conclusion IVUS could assist evaluating lumen and plaque parameters of stenotic renal arteries,guiding stent release and real-timely monitoring the effect of endovascular therapy.

【Objective:】 To analyze and explore the key points of the ethical review of real-world research in pediatric population, and to provide reference for ethical review of real-world research in pediatric population. 【Methods:】 According to the characteristics of real-world research and pediatric clinical trials, the review points of real-world research in pediatric population were analyzed and discussed in comparison with the principles and focus of ethical review in general clinical research. 【Results:】 The ethics committee should pay particular attention to the review of informed consent, privacy protection, risk benefit assessment, cost and compensation, and should also take into account the research design, data governance, research conflicts of interest, research registration and publication, etc., and conduct scientific and reasonable ethical review of real-world research in pediatric population. 【Conclusion:】 Clinical trials in pediatric population should have stricter and scientific ethical review, which can not only protect the interests of vulnerable groups of minors, but also standardize real-world research in pediatric population and promote the healthy development of pediatric clinical research, so as to better protect children and promote their health.

Objective:To analyze the clinical characteristics of acute ischemic stroke (AIS) patients with negative cerebral CT perfusion (CTP) and influencing factors for their prognoses.Methods:A retrospective analysis was performed; 448 patients with AIS admitted to Department of Neurology, Sixth Affiliated Hospital of Nantong University from January 2020 to June 2021 were enrolled. CTP images of these patients were processed by RAPID software, and they were divided into CTP-negative group and CTP-positive group according to cerebral infarction core and ischemic penumbra volumes. The clinical data were compared between patients from CTP-negative group and CTP-positive group and between patients from CTP-negative and CTP-positive subgroups accepted thrombolytic therapy. According to the prognoses 3 months after discharge, CTP negative patients were divided into poor prognosis group and good prognosis group. Independent influencing factors for poor prognosis in negative CTP patients were analyzed by univariate and multivariate Logistic regressions.Results:(1) In these 448 patients, 154 (34.4%) were with negative CTP and 294 (65.6%) were with positive CTP; compared with the CTP-positive group, the CTP-negative group had significantly younger age, significantly higher percentage of patients with diabetes, significantly lower percentage of patients with atrial fibrillation, statistically higher baseline systolic blood pressure, and significantly lower baseline National Institutes of Health Stroke Scale (NIHSS) scores, early neurological deterioration (END) incidence, modified Rankin scale (mRS) scores 3 months after discharge, and proportion of patients with poor prognosis ( P<0.05); significant differences in distributions of responsible circulations for the lesions and etiological classification (TOAST) were noted between the 2 groups ( P<0.05). Of the 448 patients, 270 received thrombolytic therapy, including 101 CTP-negative patients and 169 CTP-positive patients; compared with the CTP-positive subgroup, the CTP-negative subgroup had significantly younger age, significantly lower percentage of patients with atrial fibrillation, statistically higher baseline systolic blood pressure, and significantly lower baseline NIHSS scores, END incidence, mRS scores 3 months after discharge, and proportion of patients with poor prognosis ( P<0.05). (2) Of the 154 CTP negative patients, 31 had poor prognosis and 123 had good prognosis. Univariate Logistic regression analysis showed that baseline blood glucose, fasting blood glucose, glycosylated hemoglobin (HbA1C), baseline NIHSS scores and fibrinogen were the influencing factors for prognoses of CTP negative patients, with significant differences ( P<0.05). Multivariate Logistic regression analysis found that NIHSS ( OR=0.827, 95% CI: 0.743-0.920, P<0.001) and HbA1 C ( OR=0.763, 95% CI: 0.609-0.956, P=0.019) were independent influencing factors for poor prognosis of CTP-negative patients. Conclusion:AIS patients with negative CTP have milder neurological impairment, better prognosis, and higher safety of receiving intravenous thrombolysis than those with positive CTP; AIS patients with negative CTP enjoying high baseline NIHSS scores and HBA1c have poor prognosis.

Objective:To investigate the influencing factors for lymph node metastasis and prognosis in stage T1 and T2 esophageal squamous cell carcinoma after radical surgery and construct nomogram prediction models.Methods:The retrospective cohort study was conducted. The clinico-pathological data of 672 patients with T1 and T2 esophageal squamous cell carcinoma who were admitted to the Affiliated Hospital of North Sichuan Medical College from January 2014 to December 2019 were collected. There were 464 males and 208 females, aged (65±8)years. All patients under-went radical esophagectomy+2 or 3 field lymph node dissection. Observation indicators: (1) lymph node dissection, metastasis and follow-up. (2) risk factors for lymph node metastasis of esophageal cancer after radical resection. (3) prognostic factors of esophageal cancer after radical resection. (4) construction and evaluation of the prediction models of lymph node metastasis and prognosis of esophageal cancer after radical resection. Follow-up was conducted using outpatient examination, telephone and internet consultations to detect survival of patients up to April 2021. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Kaplan-Meier method was used to calculate survival rate and draw survival curve. Log-Rank test was used for survival analysis. Logistic regression model was used for univariate and multivariate analyses of risk for lymph node metastasis, and COX regression model was used for univariate and multivariate analyses of prognosis. Based on the results of multi-variate analysis, the nomogram prediction models for lymph node metastasis and prognosis predic-tion were constructed. The prediction discrimination of the nomogram models were evaluated using the area under curve (AUC) of the receiver operating characteristic curve (ROC). The calibration curve was used to evaluate the prediction consistency of the models. Results:(1) Lymph node dissection, metastasis and follow-up. The number of lymph node dissected was 14±8 and the number of lymph node metastasis was 2(range, 1?19) in 672 patients. Of the 672 patients, there were 182 cases had lymph node metastasis, including 58 cases in T1 stage and 124 cases in T2 stage. All 672 patients were followed up for 38 (range, 1?85)months. The average overall survival time of 672 patients was 65 months, with the 1-, 3-, 5-year overall survival rate as 89.0%, 74.3%, 66.0%, respectively. The average overall survival time of 325 patients in T1 stage and 347 patients in T2 stage were 70 months and 61 months. The 1-, 3-, 5-year overall survival rate of 325 patients in T1 stage and 347 patients in T2 stage were 95.0%, 83.5%, 73.4% and 87.4%, 69.9%, 59.2%, respectively, showing a significant difference in survival between them ( χ2=14.51, P<0.05). (2) Risk factors for lymph node metastasis of esophageal cancer after radical resection. Results of univariate analysis showed that tumor location, tumor histological grade, tumor T staging were related factors affecting lymph node metastasis of esophageal cancer after radical resection ( odds ratio=1.40, 1.54, 2.56, 95% confidence interval as 1.07?1.85, 1.20?1.99, 1.79-3.67, P<0.05). Results of multivariate analysis showed that tumor location, tumor histological grade, tumor T staging were independent factors affecting lymph node metastasis ( odds ratio=1.42, 1.61, 2.63, 95% confidence interval as 1.07?1.89, 1.25?2.09, 1.82?3.78, P<0.05). (3) Prognostic factors of esophageal cancer after radical resection. Results of univariate analysis showed that preoperative comorbidities, postoperative complications, tumor histological grade (G3), tumor T staging, tumor N staging (N1 stage, N2 stage, N3 stage), tumor TNM staging (Ⅲ stage, Ⅳ stage) were related factors affecting prognosis of esophageal cancer after radical resection ( hazard ratio= 1.48, 1.64, 2.23, 1.85, 2.09, 4.48, 4.97, 3.54, 5.53, 95% confidence interval as 1.08?2.03, 1.20?2.23, 1.47?3.39, 1.34?2.54, 1.44?3.04, 2.89?6.95, 1.57?15.73, 2.48?5.05, 1.73?17.68, P<0.05). Results of multivariate analysis showed that preoperative comorbidities, G3 of tumor histological grade, T2 stage of tumor T staging, N1 stage, N2 stage, N3 stage of tumor N staging were independent risk factors affecting prognosis of esophageal cancer after radical resection ( hazard ratio=1.57, 1.89, 1.63, 1.71, 3.72, 3.90, 95% confidence interval as 1.14?2.16, 1.23?2.91, 1.17?2.26, 1.16?2.51, 2.37?5.83, 1.22?12.45, P<0.05). (4) Construction and evaluation of the prediction models of lymph node metastasis and prognosis of esophageal cancer after radical resection. Based on the results of multivariate analysis, tumor location, tumor histological grade, tumor T staging were applied to construct a nomo-gram model for lymph node metastasis prediction of esophageal cancer after radical resection, the score of tumor location, tumor histological grade, tumor T staging were 82, 100, 100, respectively, and the sum of the scores corresponding to the lymph node metastasis rate. Preoperative comor-bidity, tumor histological grade, tumor T staging, tumor N staging were applied to construct a nomo-gram model for 1-, 3-, 5-year overall survival rate prediction of esophageal cancer after radical resection, the score of preoperative comorbidity, tumor histological grade, tumor T staging, tumor N staging were 23, 38, 27, 100, respectively, and the sum of the scores corres-ponding to the 1-, 3-, 5-year overall survival rate. Results of ROC showed that the AUC of nomogram model for lymph node metastasis prediction after radical esophagectomy was 0.66 (95% confidence interval as 0.62?0.71, P<0.05). The AUC of nomogram model for 1-, 3-, 5-year overall survival rate prediction after radical esophagectomy were 0.73, 0.74, 0.71 (95% confidence intervals as 0.66?0.80, 0.68?0.79, 0.65?0.78, P<0.05). Results of calibration curve showed that the predicted lymph node metastasis rate and the predicted 1-, 3-, 5-year overall survival rate by nomogram models were consistent with the actual lymph node metastasis rate and 1-, 3-, 5-year overall survival rate. Conclusions:Tumor location, tumor histological grade, tumor T staging are independent factors affecting lymph node metastasis in T1 and T2 esophageal squamous cell carcinoma after radical surgery and nomogram model constructed by these indicators can predict the lymph node metas-tasis rate. Preoperative comor-bidities, G3 of tumor histological grade, T2 stage of tumor T staging, N1 stage, N2 stage, N3 stage of tumor N staging are independent risk factors affecting prognosis and nomogram model constructed by these indicators can predict the overall survival rate of patients after surgery.

Pancreatic neuroendocrine neoplasms (pNENs) are highly heterogeneous, and the management of pNENs patients can be intractable. To address this challenge, an expert committee was established on behalf of the Chinese Pancreatic Surgery Association, Chinese Society of Surgery, Chinese Medical Association, which consisted of surgical oncologists, gastroenterologists, medical oncologists, endocrinologists, radiologists, pathologists, and nuclear medicine specialists. By reviewing the important issues regarding the diagnosis and treatment of pNENs, the committee concluded evidence-based statements and recommendations in this article, in order to further improve the management of pNENs patients in China.

Pancreatic neuroendocrine neoplasms (pNENs) are highly heterogeneous, and the management of pNENs patients can be intractable. To address this challenge, an expert committee was established on behalf of the Group of Pancreatic Surgery, Chinese Society of Surgery, Chinese Medical Association, which consisted of surgical oncologists, gastroenterologists, medical oncologists, endocrinologists, radiologists, pathologists, and nuclear medicine specialists. By reviewing the important issues regarding the diagnosis and treatment of pNENs, the committee concluded evidence-based statements and recommendations in this article, in order to further improve the management of pNENs patients in China.

Pancreatic neuroendocrine neoplasms (pNENs) are highly heterogeneous, and the management of pNENs patients can be intractable. To address this challenge, an expert committee was established on behalf of the Group of Pancreatic Surgery, Chinese Society of Surgery, Chinese Medical Association, which consisted of surgical oncologists, gastroenterologists, medical oncologists, endocrinologists, radiologists, pathologists, and nuclear medicine specialists. By reviewing the important issues regarding the diagnosis and treatment of pNENs, the committee concluded evidence-based statements and recommendations in this article, in order to further improve the management of pNENs patients in China.

Objective:To evaluate the current status of the registered pediatric drug or vaccine clinical trials in China for the purpose of providing a reference for the development of pediatric clinical trials in China.Methods:We collected the data about registered pediatric clinical trials that were conducted from September 6, 2013(Mandatory registration start date) to September 6, 2019 (Cut-off date) at Chinadrugtrials.org.cn platform. The survey items included trial name and number, drug classification, sponsor′s information, current trial status, completion status, etc. The clinical trials were categorized by drug group (includes chemical medicine, traditional Chinese medicine and natural medicine, biological products) and by vaccine group.Results:During the six years 349 pediatric clinical trials were registered on the platform, including 162 pediatric drug trials and 187 vaccine trials. The numbers of chemical drugs and biological products registered in 2018 were 23 and 11, respectively, the highest in the history. The number of pediatric clinical trials of traditional Chinese medicine and natural medicine was 11 in 2014, but from 2015 to 2018 only 2 to 4 trials were registered each year. The overall completion rates of the registered drug and vaccine clinical trials were 22.8% (37/162) and 41.7%(78/187), respectively. Only 42 international multicenter pediatric clinical trial projects were registered on the platform. The numbers of drug and vaccine phase Ⅰ clinical trials were 4 and 46, respectively. Thirty-six pediatric endocrine system agent clinical trials were carried out, with the largest number of all the drug categories registered on the platform.Conclusions:In recent years the number of registered pediatric drug and vaccine clinical trials increased in China. However, the number is still very limited. It is urgent to further promote the development of pediatric clinical trials.