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 evaluate the clinical efficacy of modified anterior subacromial approach plate internal fixation for three- or four-part valgus impacted proximal humeral fractures.Methods:A retrospective analysis of 35 patients treated between November 2018 and November 2021 at Fuzhou Second General Hospital was performed, including 15 males and 20 females aged 61.7±7.8 years (range: 40 to 73 years). Patients were classified under the Neer system; 17 had 3-part fractures and 18 had 4-part fractures. The modified approach accessed the fracture site via the natural interval of the deltoid anterior bundle, facilitating fracture reduction and fixation using a plate. Operative time, incision length, intraoperative fluoroscopy time, follow-up duration, Constant-Murley score, fracture healing time, visual analogue scale (VAS) for pain, and humeral neck-shaft angle were assessed. Intraoperative and postoperative complications were also recorded.Results:All patients underwent successful surgery, with an average incision length of 8.1±0.3 cm (range, 7.6-9.0 cm) and intraoperative fluoroscopy time of 6.6±0.3 seconds (3-part fractures: 6.3±0.2 s, 4-part fractures: 6.8±0.2 s, t=6.350, P<0.001). Follow-up averaged 22.1±5.8 months (range, 14-31 months). Fracture healing occurred in 11.8±1.4 weeks (range, 10-15 weeks). At the final assessment, the VAS score was 1.6±0.7 (range, 1-3), the Constant-Murley score was 89.6±2.9 (range, 84-95), and the humeral neck-shaft angle was 133.4°±3.1° (range, 128°-138°; 3-part fractures: 133.6°±3.5°, 4-part fractures: 133.3°±2.8°, t=0.288, P=0.075). No complications such as avascular necrosis of the humeral head, varus collapse of the fracture site, or axillary nerve injury were recorded. Conclusion:The modified anterior subacromial approach plate internal fixation is a minimally invasive, safe, and effective treatment for valgus impacted three- and four-part proximal humeral fractures, demonstrated by excellent surgical outcomes and absence of major complications.

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.

Abstract OBJECTIVE To investigate the effect of infliximab through concentration and antibody monitoring on the clinical outcome of children with Crohn's disease after 54 weeks of treatment. METHODS A retrospective analysis was conducted with clinical data of pediatric patients aged 6-17 years who were diagnosed with Crohn's disease at Children's Hospital, Zhejiang University School of Medicine from August 2017 to March 2023. They were divided into a reactive and a proactive monitoring group according to the monitoring method. The mucosal healing rate, disease activity, and laboratory indicators were compared after 54 weeks of treatment. RESULTS There were 77 pediatric patients with Crohn's disease included, with 34 patients from the reactive therapeutic drug monitoring group and 43 from the proactive therapeutic drug monitoring group, including 48 males and 29 females. At 54 weeks, the mucosal healing rate in the proactive therapeutic drug monitoring group was higher than that in the reactive therapeutic drug monitoring group, which was 80%(24/30) and 46.43%(13/28), respectively. The two groups had a statistical difference(P=0.01). The total clinical remission rate at 54 weeks was 84.42%(65/77), while the clinical remission rates at 54 weeks were 76.47%(26/34) in the reactive therapeutic drug monitoring group and 90.70%(39/43) in the proactive therapeutic drug monitoring group, respectively. The two groups had no statistical difference. The improvement of hypersensitive C-reactive protein, erythrocyte sedimentation rate, and serum albumin level in the proactive monitoring group was greater than in the reactive monitoring group. There was no statistical difference in the production rate of antibodies to infliximab between the two groups. CONCLUSION Proactive therapeutic drug monitoring in detecting through concentration of infliximab and antibodies may improve the mucosal healing rate compared with reactive therapeutic drug monitoring after 54 weeks of infliximab treatment.

Objective:To explore the incidence and the risk factors of post-polypectomy bleeding and polyp recurrence after colonoscopic high-frequency electrocoagulation snare polypectomy.Methods:Clinical data of 1 826 children who underwent colonoscopic high-frequency electrocoagulation snare polypectomy in the Children′s Hospital, Zhejiang University School of Medicine from January 2009 to December 2020 was retrospectively analyzed. Demographic characteristics, endoscopic manifestations, pathological features, diagnosis, occurrence of post-polypectomy bleeding and polyp recurrence were collected. The associated risk factors were analyzed by Logistic regression.Results:A total of 1 826 children (1 191 males and 635 females) with 1 967 polypectomies were included. The age was 4.6 (3.2, 6.4) years at initial diagnosis. According to the initial colonoscopy, 1 611 children (88.2%) had solitary polyps, 1 707 children (93.5%) had pedicled polyps, 1 151 children (63.0%) had polyps involving the rectum, and 1 757 children (96.2%) had hamartomatous polyps. Polyposis syndromes were diagnosed in 73 children (4.0%). The post-polypectomy bleeding occurrence was 3.8% (75/1 967). Polyps recurred in 88 children (4.8%). Girls ( OR=2.01, 95% CI 1.26-3.23) and sessile polyps ( OR=2.28, 95% CI 1.15-4.49) were risk factors for post-polypectomy bleeding (both P<0.05). Multiple polyps ( OR=17.49, 95% CI 9.82-31.18), right-colon involvement ( OR=3.44, 95% CI 1.89-6.26) and non-hamartoma ( OR=2.51, 95% CI 1.04-6.07) were risk factors for polyp recurrence (all P<0.05). Conclusions:Colonoscopic high-frequency electrocoagulation snare polypectomy has low incidence of post-polypectomy bleeding and polyp recurrence. Female patients and sessile polyps have higher risk for post-polypectomy bleeding. Multiple polyps, right-colon involvement and non-hamartoma polyps increase the risk for polyp recurrence.

Objective:To investigate radiation-induced somatic mutations and variations and provide theoretical basis for clarifying radiation-induced genetic changes and long-term effects by whole-genome sequencing analysis of the genetic variations of the victim of the " 5.7" 192Ir radiation accident in Nanjing. Methods:Normal back skin tissue, irradiated bone and soft tissues, and peripheral blood were collected from the victim 2 047 days post-irradiation. DNA of these samples was extracted and sequenced with high-throughput genomics and analyzed by bioinformatics method. The genetic variations of between irradiated and normal tissues were compared.Results:Compared with normal back skin tissue, there are large amounts of genetic variations in the irradiated bone and soft tissues and peripheral blood, including base substitution (transition, transversion), small insertion, small deletion, copy number variation (gain, loss) and structure variation (large deletion, large duplication, inversion, intra-chromosomal translocation, inter-chromosomal translocation). There were 10 666 genetic variations in the irradiated bone and soft tissues and 11 233 genetic variations in peripheral blood, where thousands of genes were involved in. These variations occurred in the exons, introns, UTR′3, UTR′5, splicing sites, within 5 kb upstream of transcription initiation site, within 5 kb downstream of transcription termination site, ncRNA and intergenic region. All chromosomes had genetic variations.Conclusions:There were a large number of genetic variations in the irradiated tissues and blood of the victim at 2 047 days after irradiation, which may affect the body function and cause the long-term effects.

Objective:To investigate the role of concurrent chemoradiotherapy in the treatment of limited-stage small cell lung cancer (LS-SCLC) and the impact of the number of chemotherapy cycle during radiotherapy (RT) on clinical prognosis.Methods:Patients with LS-SCLC treated with definitive radiotherapy from May, 2008 to September, 2016 were included in the study. The primary endpoint was overall survival (OS), which was calculated from the start of treatment to the date of death or last follow-up. The effect of the number of concurrent chemotherapy cycle and other clinical factors on clinical efficacy was analyzed. Survival analysis was performed with Kaplan- Meier method, and multivariate analysis was performed with Cox regression model. Results:Three hundred and seventeen patients were eligible for the analysis. Among them, 129 patients received sequential chemoradiotherapy and 188 patients received concurrent chemoradiotherapy. Among patients receiving concurrent chemoradiotherapy, 86 patients received 1 cycle of concurrent chemotherapy and 102 cases of 2 cycles of concurrent chemotherapy. The median follow-up time was 22.47 months. Multivariate survival analysis showed that only clinical stage, timing of RT administration and prophylactic cranial irradiation (PCI) were the independent prognostic factor for OS. The median OS in patients who received 1 cycle and 2 cycles of concurrent chemotherapy during RT were 33.8 months and 30.4 months ( P=0.400). No matter in elder patients or in younger patients, in early RT group or in late RT group and application of PCI or not, the number of concurrent chemotherapy cycle exerted no significant impact on OS. The incidence of grade 3 or above adverse events was 20% in the 1-cycle concurrent chemotherapy group, and 13.7% in the 2-cycle concurrent chemotherapy group. Conclusions:Concurrent chemoradiotherapy is the standard treatment of LS-SCLC. Two cycles of concurrent chemotherapy during RT is not necessarily superior to 1 cycle of concurrent chemotherapy. The optimal number of concurrent chemotherapy cycle during RT need to be studied in a large prospective randomized clinical trial.

Objective:The purpose of this study was to investigate the association of insomnia with social factors, vascular risk factors and neurological status in patients with cerebral infarction.Methods:217 patients with first onset cerebral infarction in Army Medical Center of PLA were enrolled in this study. According to the athens insomnia scale (AIS) score, the patients were divided into cerebral infarction insomnia group (total score >6 points) and cerebral infarction non insomnia group (total score ≤6 points). The correlation between the occurrence of insomnia in patients with cerebral infarction and humanistic factors, vascular risk factors and cerebral infarction characteristics was analyzed.Results:The incidence of insomnia in patients with cerebral infarction was 61.8%(134/217). There were significant differences in age, education level, monthly income, occupational status, depression, hypertension, diabetes mellitus, vascular control area, infarction site, National Institutes of Health Stroke Scale (NIHSS) score, activities of daily living [Barthel Index (BI) score] and aphasia between insomnia group and non insomnia group ( P<0.05); Multivariate logistic regression analysis showed that age, depression, NIHSS score, BI score and aphasia were independently associated with insomnia after cerebral infarction ( P<0.05). Conclusions:Age, depression, NIHSS score, BI score and aphasia were independent risk factors for insomnia in patients with cerebral infarction.

Objective: To evaluate the parabiotic tissue protection concept in the repairment of acute radiation-induced skin injury. Methods: Seven patients(3 males and 4 females) with acute radiation injury treated in the Department of Plastic Surgery of the Second Affiliated Hospital of Soochow University from February 2014 to January 2018. The ages of patients ranged from 45 to 76 years. The wound areas include perineum and buttock (n=3), chest(n=2), and thigh(n=2). In the early stage, subregional " sandwich" surgical dressing was used to protect the probiotic tissue. Two months later, the necrotic tissue was clearly demarcated, the debridement was underwent, and the parabiotic tissue was preserved as far as possible. Vacuum sealing drainage(VSD)was applied to cover and soak wound with normal saline to moisturize the wound and promote the benign transformation of ecological tissue. Ten days later, the granulation grown well, and the skin flaps and myocutaneous flaps with good blood supply were designed to repair the wounds. The VSD device was continued to be used, to drain effusion under flap and promote the growth of cystic cavity granulation, with the purpose to promote blood supply of the skin flap, perform the final biological cleaning effect on the parabiotic tissue of the wound surface, promote the benign transformation of parabiotic tissue, and reduce the further necrosis. Results: Seven patients with Ⅳ degree acute radiation-induced injury wounds were treated 6-10 weeks for surgery preparation, and 2-4 weeks for VSD-application after debridement. Except for part of flap was necrotized on 10th day after the first operation in one patient, all the other patients achieved satisfied outcome in a surgery. There was no further radiation-induced ulcer occurred during the 0.5-3 years of follow-up. Conclusions The concept of parabiotic tissue protection during preoperative, intraoperative and postoperative recovery phase can promote parabiotic tissue transformed to a good result after acute radiation injury, and reduce the size and depth of soft tissue necrosis, which can provide a good foundation for the secondary repair with flap and reduce complications.

Objective: To investigate the effect of down-regulation of miR-221 on cell proliferation and apoptosis of chronic myeloid leukemia (CML) K562 cells and its related regulatory mechanism. Methods: K562 cells were divided into control group, miRNAnegative control (miR-NC) group, miR-221 inhibitor group, miR-221 inhibitor+ negative control siRNA(NC siRNA) group and miR-221 inhibitor+SOCS3 siRNA group. The cells in the control group received no additional treatment. Cells in miR-NC group and miR-221 inhibitor group were transfected with miR-NC and miR-221 inhibitor, respectively. Cells in miR-221 inhibitor+NC siRNA group and miR-221 inhibitor+SOCS3 siRNA group were transfected with NC siRNA and SOCS3 siRNA, respectively, on the basis of successful transfection with miR-221 inhibitor. The transfection efficiency of miR-221 inhibitor was identified by qPCR. Cell viability in each group was measured by CCK-8 assay. Apoptosis in each group was detected by Annexin V-FITC/PI staining using a flow cytometry. The protein expressions of SOCS3, p-JAK1, p-JAK2, p-STAT3 and survivin in each group were detected by WB. Results: Compared with the control group, miR-221 expression was significantly down-regulated in miR-221 inhibitor group (P<0.01), cell viability was significantly reduced at 48 and 72 h after transfection (P<0.05 or P<0.01), the number of apoptotic cells was significantly increased (P< 0.01), the expression of SOCS3 was significantly increased (P<0.01) and the expression levels of p-JAK1, p-JAK2, p-STAT3 and survivin were significantly reduced (all P<0.01). Compared with miR-221 inhibitor group, cell viability was significantly increased at 24, 48 and 72 h after transfection (P<0.05 or P<0.01), the number of apoptotic cells was significantly decreased (P<0.01) and the expression levels of p-JAK1, p-JAK2, p-STAT3 and survivin were significantly increased in miR-221 inhibitor+SOCS3 siRNA group (all P< 0.01). Conclusion: Down-regulation of miR-221 inhibits proliferation and promotes apoptosis of K562 cells, the mechanism of which may be related with up-regulating SOCS3 expression to suppress JAK-STAT3 signaling pathway.