BACKGROUND: Regulatory dendritic cell (DCreg) subset exhibits a unique capacity for inducing immune tolerance among the variety subsets of dendritic cells (DCs) within gut-associated lymphoid tissues (GALTs). Fms-like tyrosine kinase 3 ligand (FLT3L) is involved in the differentiation of DCregs and the subsequent expansion of regulatory T-cells (Tregs) mediated by DCregs, though the precise mechanism remains poorly understood. This study aimed to explore the expansion mechanism of Treg induced by DCreg and the role of FLT3L in this process. METHODS: DCregs were distinguished from other DC subsets isolated from GALTs of BALB/c mice through a mixed lymphocyte reaction assay. The functions and mechanisms by which FLT3L promoted Treg expansion via DCregs were investigated in vitro through co-culture experiments involving DCregs and either CD4 + CD25 - T-cells or CD4 + CD25 + T-cells. Additionally, an in vivo experiment was conducted using a dextran sulfate sodium (DSS)-induced colitis model in mice. RESULTS: CD103 + CD11b + DC exhibited DCreg-like functionality and was identified as DCreg for subsequent investigation. Analysis of Foxp3 + Treg percentages within a co-culture system of CD4 + CD25 - T-cells and DCregs, with or without FLT3L, demonstrated the involvement of the FLT3/FLT3L axis in driving the differentiation of precursor T-cells into Foxp3 + Tregs induced by DCregs. Cell migration and co-culture assays revealed that the FLT3/FLT3L axis enhanced DCreg migration toward Tregs via the Rho pathway. Additionally, it was observed that DCregs could promote Treg proliferation through the Notch pathway, as inhibition of Notch signaling by DAPT (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester) suppressed Treg expansion within the co-culture system of DCregs and CD4 + T-cells or CD4 + CD25 + T-cells. Furthermore, the FLT3/FLT3L axis influenced JAG1 expression in DCregs, indirectly modulating Treg expansion. In vivo experiments further established that FLT3L promoted DCreg expansion and restored Treg balance in DSS-induced colitis models, thereby ameliorating colitis symptoms in mice. CONCLUSION The FLT3/FLT3L axis is integral to the maintenance of DCreg function in Treg expansion.
Animals ; T-Lymphocytes, Regulatory/immunology* ; Dendritic Cells/immunology* ; Mice ; Mice, Inbred BALB C ; Membrane Proteins/metabolism* ; Receptors, Notch/metabolism* ; Lymphoid Tissue/metabolism* ; Signal Transduction/physiology* ; Coculture Techniques ; Flow Cytometry

Objective:To construct a machine-learning model for predicting the progression of pancreatic cystic lesions (PCLs) based on clinical and CT features, and to evaluate its predictive performance in internal/external testing cohorts.Methods:Baseline clinical and radiological data of 200 PCLs in 177 patients undergoing abdominal thin slice enhanced CT examination at Peking Union Medical College Hospital from July 2014 to December 2022 were retrospectively collected. PCLs were divided into progressive and non-progressive groups according to whether the signs indicated for surgery by the guidelines of the European study group on PCLs were present during three-year follow-up. 200 PCLs were randomly divided into training (150 PCLs) and internal testing cohorts (50 PCLs) at the ratio of 1∶3. 15 PCLs in 14 patients at Jinling Affiliated Hospital of Medical School of Nanjing University from October 2011 to May 2020 were enrolled as external testing cohort. The clinical and CT radiological features were recorded. Multiple feature selection methods and machine-learning models were implemented and combined to identify the optimal machine-learning model based on the 10-fold cross-validation method. Receiver operating characteristics (ROC) curve was drawn and area under curve (AUC) was calculated. The model with the highest AUC was determined as the optimal model. The optimal model's predictive performance was evaluated on testing cohort by calculating AUC, sensitivity, specificity and accuracy. Permutation importance was used to assess the importance of optimal model features. Calibration curves of the optimal model were established to evaluate the model's clinical applicability by Hosmer-Lemeshow test.Results:In training and internal testing cohorts, the progressive and non-progressive groups were significantly different on history of pancreatitis, lesions size, main pancreatic duct diameter and dilation, thick cyst wall, presence of septation and thick septation (all P value <0.05) In internal testing cohort, the two groups were significantly different on gender, lesion calcification and pancreatic atrophy (all P value <0.05). In external testing cohort, the two groups were significantly different on lesions size and pancreatic duct dilation (both P<0.05). The support vector machine (SVM) model based on five features selected by F test (lesion size, thick cyst wall, history of pancreatitis, main pancreatic duct diameter and dilation) achieved the highest AUC of 0.899 during cross-validation. SVM model for predicting the progression of PCLs demonstrated an AUC of 0.909, sensitivity of 82.4%, specificity of 72.7%, and accuracy of 76.0% in the internal testing cohort, and 0.944, 100%, 77.8%, and 86.7% in the external testing cohort. Calibration curved showed that the predicted probability by the model was comparable to the real progression of PCLs. Hosmer-Lemeshow goodness-of-fit test affirmed the model's consistency with actual PCLs progression in testing cohorts. Conclusions:The SVM model based on clinical and CT features can help doctors predict the PCLs progression within three-year follow-up, thus achieving efficient patient management and rational allocation of medical resource.

Objective To demonstrate the changes in flexor digitorum and extensor digitorum tension in the affected hands with shear-wave elastography (SWE) before and after manual digitorum sensory stimulation (MDSS) in hemiplegic patients with stroke. Methods A total of 51 hemiplegic post-stroke inpatients in the Department of Rehabilitation Medicine in Second Hospital of Anhui Medical University from April to June, 2020, underwent MDSS completed by a researcher who used a bare thumb and index finger to squeeze each nail bed as well as the sides of each fingertip in the affected hand. The stimulation intensity was the minimum that could cause finger extension without obvious pain, and the interval between two stimulations was five to ten seconds. Muscular tension of the flexor digitorum superficialis, flexor digitorum profundus, flexor pollicis longus and extensor digitorum were assessed with modified Ashworth Scale (MAS) and shear-wave velocity (SWV) of SWE on the affected side before and immediately after MDSS. MAS score was -1 as low muscular tension. Results The MAS scores of all the muscles significantly reduced after MDSS (|Z| > 2.843, P < 0.001); while the changes of SWV were not significantly in all the muscles with initially MAS score of 0 or -1 (t < 1.052, P > 0.05), and it reduced in those muscles with initial MAS scores of one to three (t > 2.672, P < 0.05). The SWV were positively correlated with the MAS scores both before and after MDSS (r > 0.334, P < 0.05). Conclusion MDSS can effectively, immediately, and safely relieves muscle spasms of the flexor digitorum and facilitate active finger extension in the affected hand for hemiplegic patients with stroke. SWE is useful for quantitatively and objectively evaluating muscular tension in the affected hand for hemiplegic patients with stroke.

OBJECTIVE: To identify the causative variants in 13 Chinese pedigrees affected with oculocutaneous albinism (OCA) so as to provide genetic counseling and prenatal diagnosis to them. METHODS: Thirteen unrelated pedigrees with clinically diagnosed OCA were collected and classified based on the manifestation of skin and eyes. With informed consent obtained from the participants, peripheral blood samples were collected from the probands and their family members for the extraction of genomic DNA. Candidate variants were screened by targeted capture and next generation sequencing, and the results were validated by Sanger sequencing. Prenatal diagnosis was provided to the families upon their subsequent pregnancies. RESULTS: Causative variants were detected in all probands, including 10 with compound heterozygotes or homozygotes for TYR gene variants and 3 with compound heterozygotes for OCA2 gene variants. Among these, two variants [TYR: c.650G>C (p.Arg217Pro) and OCA2: c.516-2A>T] were unreported previously. The pathogenicity of the novel TYR: c.650G>C (p.Arg217Pro) variant was verified through bioinformatic analysis and prediction of three dimensional structure of the protein. Prenatal diagnosis was provided to 6 fetuses with a high risk for OCA. Four fetuses were found to be carriers, one did not carry the variants of the proband, and one was affected with OCA. CONCLUSION Identification of the pathogenic variants in the 13 probands, including 2 novel ones, has expanded the mutational spectrum of OCA and enabled genetic counseling and prenatal diagnosis for the families.
Albinism, Oculocutaneous/genetics* ; China ; Female ; Genetic Testing ; Humans ; Membrane Transport Proteins/genetics* ; Monophenol Monooxygenase/genetics* ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis

ObjectiveTo explore the effect of early individualized rehabilitation on patients with severe mechanical ventilation. MethodsA total of 36 patients on mechanical ventilation admitted to the ICU of the Second Affiliated Hospital of Anhui Medical University from March, 2019 to February, 2020 were randomly divided into control group (n = 18) and rehabilitation group (n = 18). All the patients completed a rehabilitation assessment within 24 hours of admission, including clinical assessment, state of consciousness and muscle strength assessment. The control group was treated with intensive care routine treatment, including symptomatic treatment of primary disease, nutritional support, placement of normal limbs, active and passive movement of limbs. The rehabilitation group received early individualized rehabilitation in addition., including active and passive limb movements, transfer training, physical factor therapy, and respiratory muscle training after the specific evaluation. The mechanical ventilation duration and ICU length of stay, the hospitalization cost, Richmond Agitation and Sedation Scale (RASS), acute Physiology and Chronic Health Evaluation Ⅱ (APACHEⅡ), and the content of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-8 were compared. ResultsThere was no significant difference in the weaning rate and hospitalization cost between two groups (P > 0.05). The mechanical ventilation duration and ICU length of stay were less in the rehabilitation group than in the control group (t > 2.067, P < 0.05). After treatment, the score of APACHEⅡ and the content of TNF-α and IL-6 decreased in the control group (t > 2.040, P < 0.05); the score of APACHEⅡ and the content of TNF-α, IL-6 and IL-8 decreased in the rehabilitation group (t > 4.141, P < 0.001); the content of TNF-α, IL-6 and IL-8 was less in the rehabilitation group than in the control group (t > 2.217, P < 0.05). The improvement of all the indexes was better in the rehabilitation group than in the control group (|Z| > 2.104, P < 0.05). ConclusionFor patients on mechanical ventilation, early individualized rehabilitation could improve the sedation, relieve the inflammatory reaction, accelerate the process of weaning, and reduce the length of stay with no extra cost.

ObjectiveTo investigate the changes of surface electromyography (sEMG) of the flexors and extensors of the affected fingers after manual digitorum sensory stimulation (MDSS) in the hemiplegic patients after stroke. MethodsFrom April to August, 2020, 50 stroke patients in Department of Rehabilitation Medicine, the Second Hospital of Anhui Medical University accepted MDSS on the nail beds and the third knuckles of affected fingers, until the fingers extended actively. The tension of affected flexor pollicis brevis, flexor digitorum superficialis and extensor digitorum were assessed with modified Ashworth Scale (MAS) before and immediately after stimulation; while the root mean square (RMS) of sEMG of bilateral flexor pollicis brevis, flexor digitorum superficialis and extensor digitorum were recorded. ResultsThe MAS score of all the muscles decreased after stimulation (|Z| > 2.699, P < 0.01), while the RMS of affected extensor digitorum increased (t = -2.069, P < 0.05). Compared with the unaffected ones, RMS of affected flexor pollicis brevis and extensor digitorum decreased before and after stimulation (t > 2.450, P < 0.05). ConclusionMDSS may immediately relieve the spasm of flexors of hemiplegic fingers after stroke, which associates with the promoting muscle strength of the extensors.

Liver fibrosis can progress to liver cirrhosis and end-stage liver disease, which may lead to poor prognosis. In addition to pathological angiogenesis and hepatic sinusoid remodeling, hepatic lymphangiogenesis also plays an important role in the progression of liver fibrosis. This article briefly describes lymphatic vessel markers and their expression in the liver, introduces the role of lymphangiogenesis in liver fibrosis, and reviews the role of liver macrophages (Kupffer cells) and lymphatic endothelial cells in lymphangiogenesis. It is pointed out lymphangiogenesis may become a potential target for the intervention of liver fibrosis, which plays an important role in the early treatment and reversal of liver fibrosis and the prevention of liver cirrhosis and end-stage liver disease.

Objective: To evaluate the effectiveness of deep learning model trained on routine CT scans when identity the malignant and benign lung nodule on target CT scans dataset. Methods: This retrospective study enrolled 923 patients with lung nodules found by chest CT scan in Shanghai Chest Hospital from January 2016 to December 2018. A total of 969 nodules with pathological report were analyzed. The deep learning based pulmonary malignant prediction method in a fine-grained classification manner was used to make the prediction, and the AUC (the area under the curve), accuracy, sensitivity and specificity of routine CT scans and target CT scans were compared, and Delong test and IDI (Integrated Discrimination Improvement) were employed to provide statistical results. Furthermore, statistical methods were used to investigate the differences between the benign and malignant classification of nodules on routine CT and on target CT. Results: In the benign and malignant discrimination task, AUC, accuracy, sensitivity and specificity on the routine scans were 0.81, 82.0%, 86.0% and 56.6% respectively, while the AUC, accuracy, sensitivity and specificity on the target scans were 0.84, 85.0%, 88.8% and 60.5% respectively. The IDI was 0.056 (Z test, P<0.05), and there was statistically significant difference in ROC (Delong test, P=0.01). Conclusions The deep learning model trained on the data set of routine CT scans can achieve better diagnostic efficiency in target CT scans data.

OBJECTIVE To identify potential mutations in two Chinese families affected with primary localized cutaneous amyloidosis. METHODS Peripheral blood samples of the family were collected with informed consent. Genomic DNA was extracted with a phenol chloroform method. All of the 17 exons and their flanking splicing sites of the OSMR gene were amplified with PCR and subjected to Sanger sequencing. Suspected mutations were verified with PCR - restriction fragment length polymorphism and high-resolution melting assays. RESULTS A missense mutation (c.1538G>A) was found in exon 10 of the OSMR gene in all of the six patients from family 1. A missense mutation (c.2081C>T) was found in exon 14 of the OSMR gene in all of the four patients from family 2. The same mutations were not found among the healthy controls. CONCLUSION Two missense mutations (c.1538G>A and c.2081C>T) were detected in the OSMR gene in two Chinese families affected with primary localized cutaneous amyloidosis. Our findings have further confirmed the pathogenicity of such mutations.

Objective We aimed to analyze the relationship among all types of HPV and cervical cancer and predict potential high-risk HPV based bioinformatics methods.Provide theoretical basis for clinical prevention and early diagnosis for cervical cancer.Methods A total of 131 types HPV gene sequences from GenBank were downloaded.HPV phylogenetic tree were constructed use the MEGA software.Results Current global HPV can be divided into five categories according to complete gene molecular evolution analysis,high-risk types HPV are height together.Further to the mucous membrane affinity types HPV gene analysis shows that the evolution of the E6,E7 gene closer to the distribution of the whole genome evolution tree.We predict the HPV 26,34,53,66,67,97 have potential dangers with cervical cancer.Conclusion Building the HPV data based on the analysis of molecular evolution method of HPV affinity relationship with cervical cancer.Provide theoretical foundations for HPV clinical prevention and treatment of cervical cancer.