Background Nano-alumina (nano-Al₂O₃) is a widely utilized nanomaterial. Its impacts on the environment and biological systems have garnered significant attention. Zebrafish serves as a common model organism in scientific research due to its high homology with the human genome and is extensively used in toxicity studies. Objective To investigate the developmental toxicity and neurotoxicity of nano-Al₂O₃ exposure in zebrafish and the corresponding mechanisms of action. Method Zebrafish embryos at 6 h post-fertilization (hpf) were randomly assigned to a control group and five dose groups exposed to nano-Al₂O₃ at concentrations of 200, 400, 600, 800, and 1000 μg·mL⁻¹, respectively. Thirty embryos were included in each group, and the culture medium was replaced every 24 h until 144 hpf. The hatching rates at 48 and 72 hpf and the cumulative malformation rates up to 144 hpf were calculated. At 144 hpf, a zebrafish behavior analyzer was used to record the movement trajectories of the zebrafish, and the total distance traveled and average speed were analyzed for each group. At 144 hpf, the development of dopaminergic neurons in transgenic zebrafish expressing vmat2: GFP, brain vessels in those expressing vegf: GFP, and central nervous system neurons in those expressing elavl3: EGFP were observed under a fluorescence microscope, and statistical analysis was conducted using Image Pro Plus. Real-time quantitative PCR was employed to detect the expression levels of neuron development-related genes (syn2α, gap43, dat), Lewy body formation-related gene (α-syn), and autophagy-related genes (pink1, parkin) at 144 hpf. Results Compared to the control group, the nano-Al₂O₃ exposed groups exhibited reduced hatching rates at 48 hpf and increased cumulative malformation rates (P<0.05), with phenomena such as delayed development, absence of the swim bladder, and body curvature. The autonomous behavioral tests revealed that the nano-Al₂O₃ exposed zebrafish showed a decrease in the total distance swum (P<0.001) and a significant reduction in average speed compared to the control group. The fluorescence observations indicated that the length of dopaminergic neurons in vmat2: GFP transgenic zebrafish was reduced in the nano-Al₂O₃ exposed groups (P<0.001). Additionally, vegf: GFP transgenic zebrafish exhibited a significant absence of brain vessels, while elavl3: EGFP transgenic zebrafish showed a weakened fluorescence intensity of central nervous system neurons (P<0.001) and a decreased length of these neurons (P<0.01). The real-time quantitative PCR analysis revealed that compared to the control group, the gene expression levels of α-syn, syn2α, dat, and gap43 were upregulated in the zebrafish exposed to nano-Al₂O₃ (except for the 400 μg·mL⁻¹ exposure group) (P<0.01), while the expression levels of parkin were downregulated in the 600 and 800 μg·mL⁻¹ nano-Al₂O₃ exposed groups, and the expression levels of pink1 were downregulated in all exposure groups except for the 200 μg·mL⁻¹ group (P<0.05). Conclusion Exposure to nano-Al₂O₃ exhibits developmental toxicity in zebrafish larvae and induces Parkinsonism-like symptoms in zebrafish. The preliminary speculation of the mechanism suggests that it may be related to nano-Al₂O₃-induced mitochondrial autophagy impairment.

Acute-on-chronic liver failure(ACLF)is a complex clinical syndrome of acute liver function deterioration on the basis of chronic liver diseases,characterized by hepatic and/or extra-hepatic organ failure and a high short-term mortality rate.At present,there is still a lack of effective treatment methods,and the mortality rate of ACLF reaches 50%-90%after comprehensive medical treatment.A simple,rapid,and accurate prognostic prediction model for ACLF can help clinicians accurately judge the prognosis of ACLF patients in the early stage,identify the patients with poor prognosis,and provide early interventions,which can improve patient prognosis to some extent and help to reduce mortality rates.With the continuous development of computer science and increasingly powerful data processing capabilities,artificial intelligence is gaining more attention and has been applied in various aspects of liver diseases including diagnosis,treatment,and prognostic prediction.With reference to the current status of research in China and globally,this article reviews the common prognostic models for ACLF and machine learning-based prognostic prediction models and summarizes the latest research advances,in order to provide new perspectives for the future development of prognostic prediction models for ACLF.

Objective To evaluate the agreement of four common HEp-2 indirect immunofluorescence assay(IFA)kits in the patients with antinuclear antibody(ANA)-associated rheumatic immune diseases(AARD)and the patients with non-autoimmune diseases(NAD).Methods The experiment in this study included two stages.In stage 1,the serum samples were randomly selected from 134 patients,and ANAs were detected by IFA at Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from Janu-ary to June 2023.All of the samples were tested using four kinds of HEp-2 IFA kits,and the consistency of qualitative results was eval-uated by statistical analysis.The kit exhibited highest positive rate was defined as Kit X.In the stageⅡ,a total of 554 serum samples(from 218 AARD and 336 NAD patients)with positive results detected by initial screening of reagent X were selected during the same period,and then the samples were tested by the other three HEp-2 IFA kits.The patterns and titers of ANA were recorded,and a semi-quantitative evaluation system was established.The reproducibility of different patterns of ANA and the consistency of the results among varying clinical characteristics,fluorescence reaction intensities and positive reaction sites in nucleus was statistically analyzed.Results There were no significant differences of qualitative results among the results from four kits(P＞0.05).The highest positive rate ap-peared in the kit m(45.86%)which was deemed as the initial screening kit X.Significant differences in the consistency of ANA pat-terns were observed.The reproducibility scores of centromeric pattern and granular pattern were higher than those of homogeneous pat-tern,dense fine speckled pattern,nuclear cytoplasmic mixed pattern and other mixed pattern with significant difference(P＜0.05).The reproducibility score of simple pattern was higher than that of mixed patterns(P＜0.05).In the nucleoplasmic region,the consistency score of the AARD group was higher than that of NAD group(P＜0.01).The consistency scores of each reaction site increased with the rise of the intensity of reaction.In the three reaction parts(nucleoplasm,nucleolus and equatorial plate),the scores between the weak and strong fluorescence reaction intensity groups showed significant differences(P＜0.001).The lowest consistency score occurred in cytoplasmic region.Conclusion The clinical interpretation for IFA ANA reports should be more cautious for the results showing weak fluorescence intensity,mixed patterns,and staining positive cytoplasmic sites.For the choice for reagents,the clinical laboratories should be also mindful of the impacts of fluorescent secondary antibodies of anti-human immunoglobulin on the test results.The develop-ment of standardized official guidelines for the manufacture of HEp-2 IFA kits should be crucial initiative for enhancing the consistency of ANA detection and promoting mutual recognition for the results between laboratories.

Objective:To explore the intervention effect of Shumu Peitu herb-partitioned moxibustion on clinical signs and symptoms and negative emotions of diarrhea-predominant irritable bowel syndrome (IBS-D) patients with liver-stagnation and spleen-deficiency pattern.Methods:A total of 72 patients with IBS-D of liver-stagnation and spleen-deficiency pattern treated in the Department of Gastroenterology of Nanjing Vniversity of Chinese Medicine from September 2021 to June 2022 were selected for randomized controlled trial. The patients were randomly divided into the observation group (2 cases dropped off, 34 cases in total) and control group (1 case dropped off, 35 cases in total) by random number table method. The patients in control group were treated with Tongxieyaofang (TXYF). The patients in observation group were treated with oral administration of TXYF and Shumu Peitu herb-partitioned moxibustion, and both groups were treated for 4 weeks. The clinical efficacy, Traditional Chinese Medicine (TCM) syndrome integral, IBS Quality of Life Questionnaire (IBS-QOL), IBS Symptom Severity Scale (IBS-SSS), Bristol Stool Form Scale and Hospital Anxiety and Depression Scale (HADS) were compared before and after treatment.Results:After treatment, the total effective rate of the observation group was 94.12%(32/34), which was higher than the 71.43%(25/35) in the control group, the difference was significant ( χ2 = 6.18, P<0.05). After treatment, the TCM syndrome integral in the observation group was (7.62 ± 4.08), which was lower than the (9.89 ± 4.71) in the control group, the difference was significant ( t = 2.14, P<0.05). After treatment of 3 days, the scores of quality of life in the five dimensions of dysthymia, behavior disorder, health worry, avoidance of eating and social function in the observation group were (82.44 ± 11.46), (80.25 ± 11.67), (76.23 ± 12.67), (59.80 ± 15.14) and (79.23 ± 11.59) points, which were different with the (73.57 ± 12.39), (72.35 ± 15.48), (69.76 ± 13.11), (50.00 ± 16.17) and (73.04 ± 13.11) points in the control group, the difference were significant ( t values were -3.09 - -2.08, all P<0.05). Three days after treatment, the score of IBS-SSS and Bristol fecal character in the observation group were (118.24 ± 40.64) and (5.09 ± 0.62) points, which were lower than the (146.86 ± 60.09) and (5.51 ± 0.66) points in the control group, the difference were significant ( t = 2.31 and 2.76, both P<0.05). After treatment, the score of HADS-A and HADS-D in the observation group were (6.26 ± 1.75) and (5.29 ± 1.47), which were different with the (7.26 ± 2.19) and (6.17 ± 2.11) in the control group, the difference were significant ( t = 2.08 and 2.00, both P<0.05). Conclusions:Shumu Peitu herb-partitioned moxibustion can effectively improve IBS-D patients with liver-stagnation and spleen-deficiency pattern, relieve clinical symptoms, reduce negative emotions, and improve quality of life.

Objective:To develop a prototype artificial intelligence immunofluorescence image recognition system for classification of antinuclear antibodies in order to meet the growing clinical requirements for an automatic readout and classification of immunof luorescence patterns for antinuclear antibody (ANA) images.Methods:Immunofluorescence images with positive results of ANA in Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from April 2020 to December 2021 were collected. Three senior technicians independently and in parallel interpreted the Immunofluorescence images to determine the ANA results. Then the images were labeled according to the ANA International Consensus on Fluorescence Patterns (ICAP) classification criteria. There were 7 labeled groups: Fine speckled, Coarse speckled, Homogeneous, nucleolar, Centromere, Nuclear dots and Nuclear envelope. Each group was randomly divided into training dataset and validation dataset at a ratio of 9∶1 by using random number table. On the deep learning framework PyTORCH 1.7, the convolutional neural network (CNN) training platform was constructed based on ResNet-34 image classification network, and the automatic ANA recognition system was established. After the model was established, the test set was set up separately, the judgment results of the model were output by ranking the prediction probability, with the results of the 2 senior technicians was taken as "golden standard". Parameters such as accuracy, precision, recall and F1-score were used as indicators to evaluate the performance of the model.Results:A total of 23138 immunofluorescence images were obtained after segmentation and annotation. A total of 7 models were trained, and the effects of different algorithms, image processing and enhancement methods on the model were compared. The ResNet-34 model with the highest accuracy andswas selected as the final model, with the classification accuracy of 93.31%, precision rate of 91%, and recall rate of 90% and F1-score of 91% in the test set. The overall coincidence rate between the model and manual interpretation was 90.05%, and the accuracy of recognition of nucleolus was the highest, with the coincidence rate reaching 100% in the test set.Conclusion:The current AI system developed based on deep learning of the ANA immunofluorescence images in the present study showed the ability to recognize ANA pattern, especially in the common, typical, simple pattern.

Objective To investigate the etiology of chronic renal failure (CRF) in middle-aged and elderly people, and to analyze related factors influencing the prognosis. Methods The clinical data of 456 middle-aged and elderly CRF patients treated in our hospital from January 2018 to January 2021 were collected. The etiology and related factors affecting prognosis were analyzed. Results The etiology of 456 middle-aged and elderly patients with CRF was as follows: 220 cases of primary glomerulonephritis (48.24%), 78 cases of diabetic nephropathy (17.14%), 65 cases of hypertensive nephropathy (14.25%), and 37 cases of pyelonephritis (8.11%), 19 cases of drug-induced renal injury (4.16%), 18 cases of obstructive nephropathy (3.95%), 12 cases of lupus nephritis (2.63%), and 7 cases of polycystic kidney disease (1.54%). The age of onset in patients with primary glomerulonephritis and lupus nephritis was lower than that in patients with other causes (P<0.05). The results showed that 94 of the 456 patients died in the hospital, and the mortality rate was 20.6%. Multivariate logistic regression analysis showed that age, severe infection, cerebrovascular disease, arrhythmia, hypertension stage III, left ventricular hypertrophy, and myocardial infarction were high-risk factors leading to poor prognosis (P<0.05). Conclusion The main causes of CRF in middle-aged and elderly patients are primary glomerulonephritis, diabetic nephropathy, and hypertensive nephropathy. Heart disease is the main factor leading to the poor prognosis of such patients. Efforts shall be put into disease prevention and control work for high-risk groups, so as to reduce the incidence and mortality of CRF.

Laboratory testing is of great value in the management of autoimmune disease. The results can help confirm a diagnosis, estimate disease severity, aid in assessing treatment effect. But the current autoimmunity laboratory system, including testing standards, quality control and supervision, does not match the national conditions well. As a result, the test reports are not mutual-recognized among laboratories. In the current background of precision medicine, with the advances of technology and the application of deep learning and artificial intelligence in the clinical laboratory field, the autoimmune laboratory has ushered in a new development trend of integration, automation and intelligence.

Objective: To explore the clinical application and effect of sideburns aesthetic reconstruction with local flaps. Methods: Between June 2012 and June 2017, there were 12 cases of patients with Benign tumor which is in the sideburn , including 7 Male patients, 5 female patients, thier ages are from 37 to 68 years old.The tumor size ranged from 0.5 cm×1.0 cm to 1.5 cm×2.5 cm, and the defects, size ranged from 1.0 cm× 1.2 cm to 1.5 cm× 3.0 cm. Single leaf flaps, double leaves flaps or local flaps which are on the ipsilateral sideburn or in the hairy area behind the ear are designed for sideburns reconstruction according to the size and shape of the defect, The flaps size ranged from 1.5 cm× 2.5 cm to 1.8 cm× 3.5 cm. Results: All flaps with 12 cases of patients survived completely with no flap blood supply obstacles, and the incision healed well, all patients were followed up for 6-24 months with no recurrence of the tumor , and the incision scars are hidden in the sideburns, And the sideburn showed satisfactory effect of aesthetic reconstruction with good symmetric cosmetic effect. Conclusions The local flaps have less injury in the surgery area , simple surgical approach, hidden incision scars, reliable blood supply and high survival rate, which do not affect the sideburns symmetry.Aesthetic result can be achieved by local flaps in the reconstruction of defect after resection of benign tumor at sideburn.

Objective To prospectively summary the piercing-out position,direction,length and piercing-in position of perforator,and investigate the feasibility of preoperative design and optimization of the anterolateral thigh flap and its clinical application.Methods All 58 cases of anterolateral thigh flaps were designed and taken from the lateral thigh area from January,2014 to January,2016.Portable Doppler ultrasound was used before an operation to detect the piercing-out position (point P) of perforators.The direction and length (lower subcutaneous segment of perforators) of perforators after leaving piercing-out position were observed during the operation.And the piercing-in positions (point P') on superficial fascia and the dermis were observed.Based on this,we added line B (anterior superior spine-lateral femoral epicondyle) and line C (anterior superior spine-the middle point of superior border of patella) in the lateral and anterior side of original ilium-patella line in the thigh (line A),respectively.Results All perforators found in 58 cases before and during the operations were located on line A or between line A and line B.No perforators were found between line A and line C.Perforators walked toward the anterior medial side after leaving the muscle membrane.The perforator vascular subcutaneous segment (distance between point P and point P') was (2.02±0.23) cm.There was rectus muscle branch in the descending branch of lateral femoral circumflex artery,while no rectus muscle cutaneous branch was seen.20 cases were designed by one-line method,12 cases were designed by two-line method,while 26 cases were designed by three-line method.Conclusion Advanced three-line method is beneficial to detect of the perforators on the anterior thigh lateral region and to reduce the intraoperative injury perforator vessels at the puncture point.Clinical application of the anterior lateral thigh flap is simple and reliable.

Objective: To investigate the therapeutic effect of advanced gourd-shaped flap with frontal branch of superficial temporal artery for defects after resection of per-auricular tumor. Methods: From June 2012 to September 2016, 9 cases with pre-auricular tumor, including 7 cases of basal cell cancer and 2 cases of squamous cell carcinoma, received extended resection operation. The tumor size ranged from 0.5 cm×1.0 cm to 1.0 cm×2.0 cm.Intraoperative frozen specimen was used to make sure no invasion at margin and defect bottom. Based on the size and location of defects, the advanced gourd-shaped flap with frontal branch of superficial temporal arterywas designed to cover the defect. The wounds at donor sites were closed directly. The flap size ranged from 2.0 cm× 5.0 cm to 3.0 cm× 7.5 cm. Results: The flaps in all the 9 cases survived completely with primary healing and no complication, like facial paralysis or abnormal feeling. The patients were followed up for 6-36 months with inconspicuous scarandno tumor relapse. Satisfactory cosmetic and functional result can be achieved. The flaps had good match in color with the surrounding tissue. Conclusions The advanced gourd-shaped flap with frontal branch of superficial temporal arteryhas a reliable blood supply and flexible transverse method with satisfactory cosmetic result. It is one of the ideal method for reconstruction of pre-auricular soft tissue defects.