OBJECTIVE To construct an evaluation index system for the core competencies of ethics committee members of drug clinical trial institution， providing a basis for optimizing the training system for committee members， improving the quality of ethical review， and fully safeguarding the safety and rights of subjects. METHODS Using methods such as literature research and expert consultation， a preliminary core competency evaluation index system was constructed. The Delphi method was employed to revise and validate it， ultimately forming an evaluation index system for the core competencies of ethics committee members. Based on this system， a questionnaire survey was conducted among 90 ethics committee members from 29 drug clinical trial institutions nationwide， comparing their importance rating and self-assessment scores of the core competency indexes. RESULTS The evaluation system constructed included 4 primary indicators （ethics and professional knowledge， ethics review ability， communication and expression ability， moral integrity and work style） and 39 secondary indicators （familiarity with the content of clinical trial-related laws and regulations， ability to complete project ethics review and identify ethical defects in research protocols within a short period of time， ability to judge the scientific value of clinical research， etc.）. The results of questionnaire survey showed that the interviewed ethics committee members had significant capability gaps in dimensions such as regulatory knowledge， ethical norms， review efficiency， risk judgment， and problem analysis. The differences between the importance rating scores of corresponding secondary indicators and the self-assessment scores were all no less than 0.38. CONCLUSIONS This study has developed a quantifiable and stratified core competency assessment tool for ethics committee members. It can provide a scientific framework for committee member training， qualification certification， and standardized management of ethics committees.

A retrospective analysis was made on the clinical data and gene mutation characteristics of 2 children admitted to the Children′s Hospital of Zhejiang University School of Medicine for epilepsy without periventricular nodules caused by the ARF1 gene mutation from August 2023 to February 2024, and relevant literature was reviewed.Both patients presented with seizures and psychomotor retardation, and 1 of them was diagnosed with West syndrome.Whole exome sequencing confirmed that the 2 patients carried a missense mutation in the ARF1 gene (c.55C>A, p.R19S).Brain magnetic resonance imaging (MRI) of 2 patients revealed no obvious abnormalities.A summary analysis of 5 cases of ARF1 gene mutations reported in three foreign literatures showed that patients with ARF1 gene mutations usually presented with seizures, developmental delay, hypotonia, mental retardation, and motor stereotypies.MRI showed periventricular nodular heterotopia, corpus callosum dysplasia, subcortical white matter abnormalities, and delayed myelination.This study found for the first time that ARF1-related disorders can occur without significant brain structural malformations, indicating that there are inconsistencies in neuroimaging findings, adding valuable phenotypic information to this gene.The differences in imaging findings may be the result of genetic background or variation in ARF1-interacting proteins, or may be caused by altered regulatory mechanisms of protein activity.

Objective:To construct a prognostic risk model for hepatocellular carcinoma(HCC),and elucidate the immune characteristics and immunotherapy response in patients with different prognostic stratification.Methods:RNA-seq data of TCGA-LIHC and ICGC(LIRI-JP),and gene microarray data of GSE14520 and GSE54236 in hepatocellular carcinoma,as well as clinical informa-tion of the corresponding samples were downloaded.First,screening of differentially expressed genes in tumor and non-tumor tissue samples from TCGA-LIHC,GSE14520 and GSE54236.For the common differential genes,univariate cox regression analysis was per-formed using TCGA-LIHC data to obtain HCC prognosis-related genes.Five genes were randomly selected as a panel,and the optimal prognostic marker panel was screened among 10 000 panels using Lasso-cox regression analysis combined with a five-fold cross-valida-tion method.TCGA-LIHC data were used as training set to construct the prognostic risk model,and ICGC data were used as validation set to test the model performance.Tumor immune dysfunction and exclusion(TIDE)algorithm and Immunophenotypic score(IPS)were used to predict immunotherapy efficacy in patients in different prognostic groups.Results:Overall survival was significantly lon-ger in low-risk group of HCC patients compared with high-risk group.Tumor proliferation rate,Treg and Th2 cell chemotaxis,stromal remodeling,and pro-tumor cytokines were significantly increased in high-risk patients,while NK cells,Th1 cells,effector cells and endothelial cells were significantly increased in low-risk patients.Immune checkpoint analysis showed that PDCD1,CTLA4 and CD276 were up-regulated in high-risk patients,while PDCD1LG2 was upregulated in low-risk patients.TIDE score and IPS results predicted that patients in low-risk group had better efficacy to immunotherapy.Conclusion:This study constructed a prognostic risk model containing three genes,DNASE1L3,RDH16 and DLGAP5,which can effectively predict the prognosis of HCC patients and assist in clinical decision making for individualized immunotherapy.

ObjectiveTo observe the clinical efficacy of Chinese medicine combined with indirect moxibustion plaster on corona virus disease 2019 （COVID-19） patients during recovery period. MethodNinety patients of COVID-19 during the recovery period were randomly divided into a Chinese medicine group， an indirect moxibustion plaster group， and a combination group，with 30 cases in each group. According to the 10th edition of COVID-19 Diagnosis and Treatment Protocol，patients in the Chinese medicine group received oral Chinese medicine based on syndrome differentiation，one dose per day， twice a day. Patients in the indirect moxibustion plaster group were treated with indirect moxibustion plaster at Zusanli （ST 36）， Pishu （BL 20）， Dazhui （GV 14）， Feishu （BL 13）， Kongzui （LU 6）， and Tiantu （CV 22），once a day，40 min each time. Patients in the combination group were treated with Chinese medicine combined with indirect moxibustion plaster. Treatment lasted two weeks. Before and after treatment，the traditional Chinese medicine （TCM） symptom score，pulmonary computed tomography （CT） score，St. George's Respiratory Questionnaire （SGRQ） score，blood routine indexes ［white blood cell count （WBC），neutrophil count （NEUT），and lymphocyte count （LYM）］， and inflammatory indexes ［C-reactive protein （CRP），serum ferritin， and interleukin-6 （IL-6）］ were observed in the three groups. The clinical efficacy was evaluated. ResultAfter treatment，the scores of TCM symptoms，pulmonary CT， and SGRQ，CRP，IL-6，and ferritin in the three groups decreased（P<0.05），while WBC and LYM increased（P<0.05）， but there was no significant difference in NEUT. The above indexes in the combination group were better than those in the other two groups（P<0.05）. After treatment， the cured and markedly effective rate was 76.7% （23/30） in the combination group， 50.0% （15/30） in the Chinese medicine group， and 46.7% （14/30） in the indirect moxibustion plaster group. The cured and markedly effective rate of the combination group was significantly higher than that of the Chinese medicine group （χ²=4.593， P<0.05） and the indirect moxibustion plaster group （χ²=5.711， P<0.05）. The total effective rate was 96.7 % （29/30） in the combination group， 93.3% （28/30） in the Chinese medicine group， and 86.7% （26/30） in the indirect moxibustion plaster group. The total effective rate of the combination group was higher than that of the Chinese medicine group and the indirect moxibustion plaster group， but the differences were not statistically significant. ConclusionChinese medicine combined with indirect moxibustion plaster can effectively improve the clinical symptoms，promote pulmonary inflammation，blood routine indexes， and inflammatory indexes， and improve the quality of life of COVID-19 patients during the recovery period，which is more advantageous than Chinese medicine alone or indirect moxibustion plaster.

ObjectiveTo observe the effect of Danshen injection （DAN） on platelet （PLT）-induced metastasis of breast cancer cells in vitro. MethodThe 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide （MTT） assay was used to observe the effect of DAN on the growth of MDA-MB-231 cells in vitro. Oris™ migration assay was used to determine the effect of DAN （final mass concentrations 4， 8， 16 g·L^-1） on PLT （1.5×10¹⁰ cells/L）-induced migration of breast cancer cells in vitro. The effect of DAN on PLT-induced cell invasion was detected by Transwell assay. Immunofluorescence and Western blot were used to detect the effect of DAN on the protein expression associated with PLT-induced epithelial-mesenchymal transition （EMT）. In addition， enzyme-linked immune-sorbent assay （ELISA） was used to determine the effect of DAN （final mass concentrations 4， 8， 16， 32， 64 g·L^-1） on the secretion of transforming growth factor-β₁ （TGF-β₁）. Western blot was used to observe the effect of DAN on the expression of podoplanin （PDPN） protein in MDA-MB-231 cells induced by PLT. ResultCompared with the blank group， the DAN groups （32 and 64 g·L^-1） showed decreased A₅₇₀ （P<0.05， P<0.01）， and there was no significant difference in A₅₇₀ between DAN groups （4， 8， 16 g·L^-1）. Compared with the blank group， the PLT group showed increased cell migration and invasion， while DAN groups significantly inhibited PLT-induced cell migration and invasion. Compared with the blank group， the PLT group showed decreased expression of E-cadherin， while DAN could significantly reverse this effect of PLT. Compared with the blank group， the PLT group showed increased Slug and Snail protein expression （P<0.05， P<0.01）， while DAN significantly reversed Snail protein expression induced by PLT （P<0.05， P<0.01）. The content of TGF-β₁ in the PLT group increased （P<0.01）， while the secretion of TGF-β₁ induced by PLT decreased in the DAN groups （16， 32， and 64 g·L^-1） （P<0.05， P<0.01）， and the secretion of TGF-β₁ was not significantly affected in other DAN groups. PDPN protein expression in the PLT group increased （P<0.01）， while DAN could significantly inhibit PLT-induced PDPN expression （P<0.01）. ConclusionDAN can inhibit PLT-induced migration， invasion， and EMT of breast cancer cells. The mechanism may be related to the direct action between breast cancer cells and tumor cells by down-regulating PDPN expression and interfering with PLT and has nothing to do with the effect of TGF-β₁ secretion of PLT.

Objective:To explore the diagnostic value of the toxicant and drug detection in clinical poisoning diseases and analyze the clinical characteristics of patients with positive poison test.Methods:This study was a multicenter retrospective cohort study. Sampling and clinical information data were collected between October 1, 2020 and September 30, 2022 from 41 tertiary hospitals in and around Jiangsu province. The clinical characteristics of patients with positive toxicology tests were analyzed, and the correlation between the drug sampling situation and the test results was analyzed..Results:A total of 895 patients with clinical diagnosis or suspected poisoning were enrolled in this study. Among them, 652 patients had positive results, accounting for 72.85%. Among all positive patients, 506 patients were exposed to a single poison and 147 patients were exposed to multiple poisons. The top three poisons were pesticide herbicides (202 cases, 30.98%), sedative and psychotropic drugs (151 cases, 23.16%), and pesticide insecticides (97 cases, 14.88%). Among 541 patients with clear exposure history, the positive rate was 78.19%, and among 354 patients with unclear exposure history, the positive rate was 64.69%. The top three poisons (drugs) of patients with unclear exposure history were sedative and psychotropic (82, 12.58%), herbicide (26, 3.99%), and rodenticide (22, 3.37%). Patients who admitted to hospital for unexplained consciousness disorder, abnormal blood coagulation function and multiple organ dysfunction were more likely to obtain positive poison test results.Conclusions:There is uncertainty in the exposure history of poisoning diseases, so it is necessary to improve the detection of toxic substances as soon as possible. Toxicant testing should be considered when patients have impaired consciousness, abnormal coagulation function and multiple organ dysfunction.

Objective:To explore the clinical characteristics of poisoned patients with poisons purchase online.Methods:A retrospective case-control study was conducted on poisoned patients purchased poisons online from 1st January 2021 to 31th May 2022 in the Emergency Department of the First Affiliated Hospital of Nanjing Medical University. The clinical data including sex, age, way of medical treatment, cause of poisoning, exposure routes, category of toxic drugs, gastric lavage, toxic detection and prognosis of patients were collected and compared with those patients obtained poisons at stores as the control group.Results:Totally 318 poisoned patients were included in this study, of which 44 (13.8%) were obtained poisons online. Compared with the patients obtained poisons at stores, the patients obtained poisons online were younger ( P<0.001), and had higher proportion of suicide intention ( P=0.006), more oral route exposure ( P=0.029), and more proportions of receiving gastric lavage before transfer to the hospital ( P=0.001). Pesticides and fertilizers with organic heterocycles were the main types of poisons in the online group, and there was no statistical difference in the distribution of poisons compared with the control group. Mixed drug poisoning was the leading cause in both online group (27.8%) and control group (38.8%) in drug overdose poisoned types, followed by dextromethorphan (16.7%) and estazolam (15.5%) in the online group. Conclusions:Young people are the main group getting poisons through the Internet. Health education should be strengthened for this group, and online shopping platforms should pay attention to the poisoning risk of potential overdose drugs or poisons transactions.

The clinical data of a case with late-onset isolated sulfite oxidase deficiency(ISOD)admitted in the Department of Neurology, Children′s Hospital, Zhejiang University School of Medicine in July 2021 were retrospectively analyzed.Fifteen previously published cases of late-onset ISOD were also reviewed.The patient was a girl, who was hospitalized because of " motor regression with mental retardation for 5 days" at 1 year old.The manifestations of the patient were extrapyramidal symptoms, regression of motor development and seizures.The level of urinary sulfites in the patient was increased.Magnetic resonance imaging (MRI) features were bilateral pallidus and substantia nigra.Gene sequencing suggested a pure missense mutation of the sulfite oxidase( SUOX) gene c. 650(exon5)G>A(p.Arg217Gln). In 16 cases of late-onset ISOD, the median age at onset and diagnosis was 10.5 months and 34.0 months, respectively.The common clinical manifestations were hypotonia (13 cases), seizures (10 cases), movement disorders (9 cases), and ectopia lentis (6 cases). The most common brain MRI feature was pallidus changes (11 cases), followed by lesions of substantia nigra (5 cases), and cerebral atrophy (4 cases). Fourteen cases of late-onset ISOD showed a positive urinary sulfite test.The missense mutation of the SUOX gene was found in 9 cases.It suggested that brain MRI involvement of bilateral pallidus, high excretion of urine sulfites and the missense mutation of the SUOX gene were important diagnostic clues for late-onset ISOD.

Objective    To investigate the role of kinase insert domain containing receptor (KDR) positive cells in the formation of cardiospheric structure, myocardium and vessels. Methods    Twenty-four Wistar rats weighting 250 g were selected. Cardiosphere-derived cells were isolated by enzymatic digestion of rat hearts, and their immunological phenotypes were analyzed by using fluorescence-activated cell sorting (FACS). The cardiomyogenic and vasculogenic potential was diagnosed by immunohistochemistry. Results    KDR positive cells grew exponentially and formed cell clusters. It also could generate myocardial precursor cells (cardiac troponin T positive). And these cells can develop spontaneous contraction activity in vitro. Meanwhile, KDR positive cells formed many vessel-like structures through a budding process. Conclusion    KDR positive cells form cardiospheric structure in vitro culture, and exhibit differentiation potential towards the cardiac and vascular cells. Therefore, KDR positive cells may have a broad prospect of clinical application as cell donors.

Objective:To construct a novel biomimetic calcium phosphate (BioCaP) scaffold loaded with bone morphogenetic protein-2 (BMP-2),and to investigate its role in the osteogenesis of human adipose-derived stem cells (hASCs) in vitro and in vivo.Methods:The BioCaP scaffold coprecipitated with BMP-2 (BMP-2-BioCaP) was constructed in this study.Field emission scanning electron microscopy (SEM) was used to analyze the morphology of the surfaces.The release kinetics was measured to evaluate the slow-release characteristics in vitro.BMP-2-BioCaP was immersed in proliferation medium (PM) or osteogenic medium (OM),respectively.The supernatants were collected and used to culture hASCs in vitro.Cell numbers were determined using the cell-counting kit-8 (CCK-8) to assess the cell proliferation.After 7 and 14 days,alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP.After 14 and 21 days,the calcification deposition was determined by alizarin red S (ARS) staining and quantification.The expressions of the osteoblast-related genes were tested on day 4 and day 14.In the in vivo study,6 nude mice were used and implanted subcutaneously into the back of the nude mice for 4 groups:(1) BioCaP scaffold only,(2) BioCaP scaffold + hASCs,(3) BMP-2-BioCaP scaffold,(4) BMP-2-BioCaP scaffold + hASCs (test group).After 4 weeks of implantation,hematoxylin-eosin (HE) staining was performed to evaluate the in vivo osteogenesis of hASCs.Results:SEM observations showed that BioCaP and BMP-2-BioCaP scaffold were entirely composed of straight,plate-like and sharp-edged crystal units,and the length of the crystal units varied between 5 and 10 μm.Release kinetics analysis demonstrated that BMP-2 incorporated with BioCaP could be released at certain concentration and last for more than 21 days,and the accumulative protein release could reach 20％.CCK-8 assays showed that cell proliferation was not significantly affected by BMP-2BioCaP.ALP activity was higher by the induction of OM + BMP-2-BioCaP than of the other groups (P ＜0.01).More mineralization deposition and more expressions of osteoblast-related genes such as Runt-related transcription factor 2 (RUNX2),ALP,osteopontin (OPN) and osteocalcin (OC) were determined in the OM + BMP-2-BioCaP group at different time points (P ＜0.01).HE staining showed that,in the test group and BMP-2-BioCaP scaffold group,the extracellular matrix (ECM) with eosinophilic staining were observed around hASCs,and newly-formed bone-like tissues could be found in ECM around the scaffold materials.Moreover,compared with the BMP-2-BioCaP scaffold group,more bone-like tissues could be observed in ECM with typical structure of bone tissue in the test groups.No obvious positive results were found in the other groups.Conclusion:BMP-2-BioCaP scaffold could achieve slow-release of BMP-2 and promote the osteogenic differentiation of hASCs in vitro and in vivo.The novel tissue-engineered bone composed of hASCs and BMP-2-BioCaPis promising for the repair of bone defect.