Background Chlorinated perfluoroalkyl ether sulfonate Cl-PFAES, trade name F-53B, a novel per- and polyfluoroalkyl substance (PFAS), has been shown to induce multi-organ toxicity in humans and cross the blood-brain barrier. However, its toxic effects and underlying mechanisms on neural stem cells (NSCs) remain unclear. Objective To investigate the impact of F-53B on NSCs proliferation and differentiation through oxidative stress and explore its potential molecular mechanisms in associations with mitochondrial function damage and the expression of autophagy-related gene (PINK1/Parkin). Methods Primary NSCs isolated from neonatal C57BL/6 mice were used as a model and exposed to F-53B at concentrations of 0, 33, or 100 μmol·L⁻¹ for 24 h. Cell viability was assessed using the cell counting kit-8 (CCK-8) assay, while proliferation was evaluated by the 5-ethynyl-2’-deoxyuridine (EdU) incorporation assay. Immunofluorescence staining was performed to observe differentiation phenotypes. Intracellular and mitochondrial reactive oxygen species (ROS) levels were quantified using dihydroethidium (DHE) and MitoSOX probes, respectively. Mitochondrial morphology was observed using MitoTracker Green. ATP level was measured with a commercial kit. Additionally, real-time quantitative polymerase chain reaction (qPCR) was conducted to quantify the expression of PINK1 and Parkin genes. Results Exposure to 100 μmol·L⁻¹ F-53B significantly reduced cell viability to 93.6% of the control group (P<0.01), and decreased the proportion of EdU⁺ cells (P<0.01), indicating proliferation inhibition. The differentiation analysis showed a reduction in neuronal generation, axonal shortening, and an increase in astrocytes. The 100 μmol·L⁻¹ F-53B exposure elevated intracellular ROS to 122% (P<0.01) and mitochondrial ROS (MitoROS) to 135% (P<0.001) of the control levels, leading to mitochondrial fragmentation. The ATP levels after the F-53B exposure decreased to 62.4% relative to the control group (P<0.001). Furthermore, the mRNA expression levels of PINK1 and Par after the F-53B exposure were notably reduced (P<0.05). Conclusion F-53B may induce oxidative stress, thereby disrupting mitochondrial morphology and function while inhibiting the PINK1/Parkin-mediated mitophagy pathway, ultimately leading to impaired neural stem cell proliferation and abnormal differentiation. This study provides new insights into the neurotoxicity mechanisms of F-53B.

BACKGROUND:The in vitro construction of intestinal smooth muscle layer,as an important component of the intestinal wall,has attracted much attention in the bionic construction of tissue-engineered intestinal canal. OBJECTIVE:To explore the effects of cyclic mechanical stretching on the growth activity of intestinal smooth muscle cells and the expression of functional genes within collagen strips. METHODS:The collagen band culture system of intestinal smooth muscle cells was constructed using a self-designed collagen strip stretching culture device with self-made rat tail collagen as a scaffold and primary rat intestinal smooth muscle cells as seed cells.EthD-1/Calcein-AM cell activity staining,magenta staining,cytoskeleton-Ki67 immunofluorescence staining were used to observe the growth activity and proliferation of the cells,and quantitative RT-PCR was used to detect the expression of desmin,α-sma,and vimentin functional genes. RESULTS AND CONCLUSION:The collagen band culture system of intestinal smooth muscle cells was successfully constructed,and intestinal smooth muscle cells in the band had good cell activity.The number of Ki67 positive cells increased and desmin,α-sma and vimentin were significantly overexpressed under cyclic stretching and dynamic culture conditions(P＜0.001).To conclude,mechanical stimulation is beneficial to maintain the growth phenotype of smooth muscle cells and promote their functional differentiation during three-dimensional culture in vitro.

Objective To investigate and analyze factors affecting depressive disorders in elderly stroke patients after rehabilitation treatment.Methods A total of 1 100 elderly stroke patients receiving rehabilitation treatment in the three hospitals(Beijing Electric Power Hospital of State Grid Corporation,Beijing Fengtai Hospital of Integrative Medicine and Beijing Daxing Hospital of Integrative Medicine)were selected as the study subjects.General data were collected,and patients were surveyed using the daily life ability scale(BI),family care scale and social support scale(SSRS),according to the Hamilton Depression Scale(HAMD)to determine whether patients developed depressive disorders.Multivariate Logistic regression analysis were used to analyze influencing factors of depressive disorders in elderly stroke patients after rehabilitation treatment.Results A total of 1 100 survey questionnaires were issued and 1 073 were retrieved,with an effective rate of 97.5%,and among them,386 cases(36.0%)were diagnosed with depressive disorder,and 687 cases(64.0%)without depressive disorder.There were significant differences in the incidence of depression between patients with different ages,complications,per capita family income,BI score,family caring degree and SSRS score(P＜0.05).Multivariate Logistic regression analysis showed that advanced age(OR=1.337,95%CI:1.014-1.763),complication(OR=3.214,95%CI:1.365-7.569),lower per capita monthly family income(OR=2.263,95%CI:1.137-4.503),lower BI score(OR=1.872,95%CI:1.158-3.026),average family caring degree(OR=2.639,95%CI:1.200-5.802)and lower SSRS score(OR=1.557,95%CI:1.044-2.322)were risk factors for depression in elderly stroke patients after rehabilitation treatment(P＜0.05).Conclusion More attention should be given to elderly stroke patients to improve family care and social support,thus reducing the risk of depressive disorders after rehabilitation.

Objective:To retrospectively analyze the treatment status of tyrosine kinase inhibitors (TKI) in newly diagnosed patients with chronic myeloid leukemia (CML) in China.Methods:Data of chronic phase (CP) and accelerated phase (AP) CML patients diagnosed from January 2006 to December 2022 from 77 centers, ≥18 years old, and receiving initial imatinib, nilotinib, dasatinib or flumatinib-therapy within 6 months after diagnosis in China with complete data were retrospectively interrogated. The choice of initial TKI, current TKI medications, treatment switch and reasons, treatment responses and outcomes as well as the variables associated with them were analyzed.Results:6 893 patients in CP ( n=6 453, 93.6%) or AP ( n=440, 6.4%) receiving initial imatinib ( n=4 906, 71.2%), nilotinib ( n=1 157, 16.8%), dasatinib ( n=298, 4.3%) or flumatinib ( n=532, 7.2%) -therapy. With the median follow-up of 43 ( IQR 22-75) months, 1 581 (22.9%) patients switched TKI due to resistance ( n=1 055, 15.3%), intolerance ( n=248, 3.6%), pursuit of better efficacy ( n=168, 2.4%), economic or other reasons ( n=110, 1.6%). The frequency of switching TKI in AP patients was significantly-higher than that in CP patients (44.1% vs 21.5%, P<0.001), and more AP patients switched TKI due to resistance than CP patients (75.3% vs 66.1%, P=0.011). Multi-variable analyses showed that male, lower HGB concentration and ELTS intermediate/high-risk cohort were associated with lower cytogenetic and molecular responses rate and poor outcomes in CP patients; higher WBC count and initial the second-generation TKI treatment, the higher response rates; Ph + ACA at diagnosis, poor PFS. However, Sokal intermediate/high-risk cohort was only significantly-associated with lower CCyR and MMR rates and the poor PFS. Lower HGB concentration and larger spleen size were significantly-associated with the lower cytogenetic and molecular response rates in AP patients; initial the second-generation TKI treatment, the higher treatment response rates; lower PLT count, higher blasts and Ph + ACA, poorer TFS; Ph + ACA, poorer OS. Conclusion:At present, the vast majority of newly-diagnosed CML-CP or AP patients could benefit from TKI treatment in the long term with the good treatment responses and survival outcomes.

OBJECTIVE: To explore the genetic basis for three Chinese patients with McCune-Albright syndrome (MAS). METHODS: Three children who had respectively presented at Shandong Provincial Hospital in April 2019 and Peking Union Medical College Hospital in August 2020 and May 2021 were selected as the research subjects. Peripheral blood samples of the probands and their family members were taken for the extraction of genomic DNA. Potential variants were screened by whole exome sequencing (WES), and candidate variants were validated by Sanger sequencing of the patients and their family members. RESULTS: The proband from family 1 was found to harbor a heterozygous c.601C>T (p.R201C) missense variant in exon 8 of the GNAS gene, whilst the probands from families 2 and 3 were both found to harbor a heterozygous c.602G>A (p.R201H) missense variant in exon 8 of the GNAS gene. Both variants were known to be pathogenic, and all probands were found to be mosaics for the corresponding variants but with various degrees. CONSLUSION WES can effectively diagnose MAS and other somatic genetic disorders. In this study, the combined WES and Sanger sequencing have verified the degree of mosaicisms of pathogenic variants in the three MAS patients, albeit no apparent correlation was found between the degree of mosaicisms and the phenotype of patients. Above finding has provided a basis for genetic counseling and prenatal diagnosis for the affected families.
Humans ; Mutation ; Fibrous Dysplasia, Polyostotic/genetics* ; East Asian People ; Exons ; Phenotype ; Pedigree

Objective To investigate the clinicopathological characteristics and prognosis of well-differentiated rectal neuroendocrine tumor(RNET).Methods A retrospective analysis was conducted using the clinical data from 83 patients with well-differentiated RNET from August 2017 to December 2021,including clinical manifestations,endoscopy,endoscopic treatment,postoperative complications,postoperative pathology,follow-up and prognosis.Pathological results according to the 2019 World Health Organization(WHO)Classification of digestive system tumors,83 patients were divided into G1 stage group(72 cases)and G2 stage group(11 cases);Based on the number of tumors in the patient,83 patients were divided into two groups:single RNET group(77 cases)and multiple RNET group(6 cases),the expressions of chromogranin A(CgA),synapsin(Syn)and CD56 were compared among different groups.Results Based on pathological findings in the group,G1 stage group CgA positive rate was significantly higher than that of G2 stage group,the difference was statistically significant(χ2 = 4.23,P = 0.040);Based on the number of tumors,multiple RNET group CgA positive rate was significantly higher than that of single RNET group,the difference was statistically significant(χ2 = 5.74,P = 0.017).It was no significant difference in Syn and CD56 between the two groups(P＞0.050).Conclusion Well-differentiated RNET has no specific clinical manifestations.It is mostly isolated in G1 stage and single RNET.ESD is safe and has a good prognosis,the positive rate of CgA is higher in G1 stage patients,and the positive rate of CgA is higher in patients with multiple RNET.

Objective:To evaluate the teaching effectiveness of independent experimental design from students' active learning behavior, and further provide the basis for advancing the reform of functional experimental teaching and teaching quality.Methods:In June 2019, 186 undergraduates (5-year-programme and 8-year-programme) of Xiangya School of Medicine were included in the teaching research. Self-administered questionnaires were applied to characterize students' active learning behavior in independent experimental design education. Spearman rank correlation analysis and Logistic regression analysis were used in the study. SPSS 23.0 was used for descriptive analysis of the data.Results:During the independent experimental design, 85.0%(158/186) of the students thought it was necessary and important to conduct independent experimental design education; 72.6%(135/186) of the students tentatively raised new scientific questions; 97.8%(182/186) of the students actively searched literature; 77.4%(144/186) of the students participated in reply positively. The value of correlation coefficient of actively learning behavior "tentatively raising new science questions" and teaching effectiveness "improving the ability of scientific thinking" was 0.81. And only 42.5%(79/186) of the students agreed that students needed to summarize after reporting.Conclusion:Independent experimental design education is welcomed and widely accepted by students, which has effectively improved the capacity for scientific research and innovation spirit of students. Whether students' active learning behavior can be fully mobilized in the education practice is closely related to the teaching effect. And the cultivation of leadership and leading consciousness still need to be improved.

Objective:To analyze the serology and molecular typing of Listeriamonocytogenes isolated from patients in Henan, and to explore the epidemic situation of listeriosis, construct the molecular traceability database of patient isolates, so as to provide laboratory basis for listeriosis traceability. Methods:From January 2015 to July 2020, 71 positive Listeriamonocytogenes cases were monitored in 16 sentinel hospitals in Henan. Eighty samples of positive cases were collected for detection, and 71 positive strains were obtained for molecular typing. According to the Entry-exit Inspection and Quarantine Industry Standard of China (SN/T 2521-2010) and the instructions for the use of diagnostic serum of Listeriamonocytogenes, 80 strains of Listeriamonocytogenes were serotyped, and PFGE cluster analysis was performed according to the User Manual of National Foodborne Disease Molecular Traceability Network. Results:A total of 71 positive listeriosis cases were detected, of which 38 cases were perinatal cases and 33 cases were non-perinatal cases. Among the 80 positive cases of listeriosis, 58.75% (47/80) were from perinatal cases, 20.00% (16/80) were from non-perinatal cases with underlying diseases, the proportion of>1 month-≤5 years old,>5-≤60 years old and >60 years old was 7.50% (6/80), 12.50% (10/80) and 1.25% (1/80), respectively, in non-perinatal age group. There were 5 types of specimens, 73.75% (59/80) were blood, 15.00% (12/80) were cerebrospinal fluid, and 3.75% (3/80) were stool, intrauterine swab and sputum. 80 strains of Listeriamonocytogenes were classified into three serotypes, Type 1/2b, Type 1/2a and Type 4b accounted for 61.25% (49/80), 35.00% (28/80) and 3.75% (3/80) respectively. The 71 strains of Listeriamonocytogenes were digested by AscⅠ, and 58 bands were obtained. Each band type included 1-4 strains, and the similarity was 60.8%-100%. GX6A16HA0005, GX6A16HA0011, GX6A16HA0030, GX6A16HA0023, GX6A16HA0029 and GX6A16HA0054 were dominant bands, including 4, 4, 4, 3, 2 and 2 strains respectively. Four strains of GX6A16HA0005 from 2016, 2018 and 2020 were isolated. One strain from 2016 and one strain from 2018 were from Puyang City. Four strains of GX6A16HA0011 were isolated from samples of 2016, 2018 and 2020, including two strains of 2020 from Luoyang City. Four strains of GX6A16HA0030 were isolated from 2018 samples from Luoyang City, Shangqiu City and Zhengzhou City, respectively. Three strains of GX6A16HA0023 were isolated from 2017 and 2018 samples, of which one strain from 2017 and one strain from 2018 were from Luoyang City. Two strains of GX6A16HA0029 were isolated from 2018 samples, from Kaifeng City and Puyang City respectively. Two strains of GX6A16HA0054 were isolated from 2020 from Pingdingshan City and Anyang City, respectively. The PFGE patterns of 4 strains with different serotypes were the same. Conclusion:Listeriosis cases in Henan are mainly found in patients during the perinatal period, and in elderly, new-born, and low immunity population. The infection type is mainly invasive infection; the serotypes of epidemic strains are 1/2a, 1/2b and 4b, and the results of PFGE typing of strains are diverse. There is a consistent phenomenon of cross-year or different regions in the same year, different time zones in the same year and the same region; phenotyping and genotyping or different genotyping techniques should be combined in the traceability analysis.

Background: In suckling piglets, transmissible gastroenteritis virus (TGEV) causes lethal diarrhea accompanied by high infection and mortality rates, leading to considerable economic losses. This study explored methods of preventing or inhibiting their production.Bovine antimicrobial peptide-13 (APB-13) has antibacterial, antiviral, and immune functions. Objectives: This study analyzed the efficacy of APB-13 against TGEV through in vivo and in vitro experiments. Methods: The effects of APB-13 toxicity and virus inhibition rate on swine testicular (ST) cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT). The impact of APB-13 on virus replication was examined through the 50% tissue culture infective dose (TCID50 ). The mRNA and protein levels were investigated by real-time quantitative polymerase chain reaction and western blot (WB). Tissue sections were used to detect intestinal morphological development. Results: The safe and effective concentration range of APB-13 on ST cells ranged from 0 to 62.5 µg/mL, and the highest viral inhibitory rate of APB-13 was 74.1%. The log10 TCID50 of 62.5 µg/mL APB-13 was 3.63 lower than that of the virus control. The mRNA and protein expression at 62.5 µg/mL APB-13 was significantly lower than that of the virus control at 24 hpi. Piglets in the APB-13 group showed significantly lower viral shedding than that in the virus control group, and the pathological tissue sections of the jejunum morphology revealed significant differences between the groups. Conclusions APB-13 exhibited good antiviral effects on TGEV invivo and in vitro.

Objective: To investigate the value of CT wavelet texture analysis based on primary tumor of papillary thyroid carcinoma (PTC) in predicting central lymph node metastasis (CLNM). Methods: A retrospective analysis was performed to 250 patients (307 nodules) who pathologically confirm with PTC in the First Affiliated Hospital of Kunming Medical University from December 2013 to August 2019. Thyroid dual phase scanning was performed in all patients within two weeks before surgery. All patients underwent central or total cervical lymph node dissection, among which 160 cases (189 nodules) were classified as training sets, while 90 cases (118 nodules) were in the verification sets. Besides, all patients were divided into CLNM group and no CLNM group according to pathology. The DeepWise software were used to manually delineate PTC primary nodules on venous phase CT images, and 576 wavelet texture features were extracted. The differences of texture feature parameters between the two groups were compared. The top 10 wavelet texture features of the area under curve (AUC) value were manually selected as the best parameters. Multivariable logistic regression analysis was used to establish and verify the model, the optimal cutoff value was found by using receiver operating characteristic curve analysis. Results: Totally 124 features were statistical difference between the two groups. The top 10 characteristic parameters for manual diagnosis with AUC values ranged from 0.599 to 0.630 (P<0.05), multi-collinearity test and multi-logstic regression analysis showed that there was no collinear correlation between the above 10 features, and small-area low-gray emphasis was an independent predictor of risk factors. The AUC value, sensitivity, specificity, and accuracy of the predictive model for the diagnosis of CLNM in the training set were 0.693, 62.84%, 60.47%, 62.96% and validation set were 0.602, 64.95%, 33.33%, and 59.32%, respectively. Conclusion Wavelet texture analysis in CT venous phase may allow detection of CLNM of PTC.