Objective:To develop a convolutional neural network model of whole slide imaging of cerebrospinal fluid cells for rapid and accurate identification and classification of tumor cells in cerebrospinal fluid.Methods:A total of 8 692 cerebrospinal fluid cytology smears from Huashan Hospital Affiliated to Fudan University from January 2nd, 2019, to December 27th, 2024. As randomly assigned, the training set included 4 941 benign and 1 745 malignant samples, while the validation set comprised of 1 368 benign and 638 malignant samples. Whole-slide digital images were acquired using a cytopathology scanner, cells (clusters) were annotated for classification, and a deep learning model was constructed via tiled image patches for cell detection and classification. Model performance was evaluated using accuracy, sensitivity, specificity, and other indicators. The classification efficiency of manual microscopy was compared.Results:The model achieved a mean precision of 96.75% for cerebrospinal fluid cell classification. For malignant tumor cells, the classification accuracy was 96.61% (mAP=98.36%, AUC=0.97). Subtype classification accuracies for epithelial/epithelioid tumors and small round cell tumors were 97.13% (AUC=0.98) and 95.58% (AUC=0.93), respectively. Compared with manual microscopy, which took (9.70±0.82) minutes for classifying 200 cells, (18.27±1.21) minutes for 500 cells, and often exceeded 60 minutes or infeasible for full slides, the AI model took (3.46±0.49) seconds for 200 cells, (6.76±0.82) seconds for 500 cells, and a median of 48.57 seconds for full slides ( P<0.001), representing an efficiency improvement of approximately 161-170 times, significantly enhancing diagnostic efficiency. Conclusion:This fully automated hierarchical deep learning model enables efficient and accurate tumor cell identification and classification in CSF, providing an effective auxiliary tool for the rapid diagnosis of meningeal carcinomatosis.

Objective:To develop a convolutional neural network model of whole slide imaging of cerebrospinal fluid cells for rapid and accurate identification and classification of tumor cells in cerebrospinal fluid.Methods:A total of 8 692 cerebrospinal fluid cytology smears from Huashan Hospital Affiliated to Fudan University from January 2nd, 2019, to December 27th, 2024. As randomly assigned, the training set included 4 941 benign and 1 745 malignant samples, while the validation set comprised of 1 368 benign and 638 malignant samples. Whole-slide digital images were acquired using a cytopathology scanner, cells (clusters) were annotated for classification, and a deep learning model was constructed via tiled image patches for cell detection and classification. Model performance was evaluated using accuracy, sensitivity, specificity, and other indicators. The classification efficiency of manual microscopy was compared.Results:The model achieved a mean precision of 96.75% for cerebrospinal fluid cell classification. For malignant tumor cells, the classification accuracy was 96.61% (mAP=98.36%, AUC=0.97). Subtype classification accuracies for epithelial/epithelioid tumors and small round cell tumors were 97.13% (AUC=0.98) and 95.58% (AUC=0.93), respectively. Compared with manual microscopy, which took (9.70±0.82) minutes for classifying 200 cells, (18.27±1.21) minutes for 500 cells, and often exceeded 60 minutes or infeasible for full slides, the AI model took (3.46±0.49) seconds for 200 cells, (6.76±0.82) seconds for 500 cells, and a median of 48.57 seconds for full slides ( P<0.001), representing an efficiency improvement of approximately 161-170 times, significantly enhancing diagnostic efficiency. Conclusion:This fully automated hierarchical deep learning model enables efficient and accurate tumor cell identification and classification in CSF, providing an effective auxiliary tool for the rapid diagnosis of meningeal carcinomatosis.

Purpose To explore the clinicopathologic fea-tures of Epstein-Barr virus-positive inflammatory follicular den-dritic cell sarcoma(EBV+IFDCS).Methods The clinico-pathologic features of 9 cases of EBV+IFDCS were retrospective-ly analyzed and followed up.Results The age of 9 patients with EBV+IFDCS ranged from 22 to 78 years(mean 44.7 years).7 cases occurred in the liver and 2 in the spleen.Fi-brinoid degeneration and hyaline degeneration in the vessel walls(6/9),eosinophilic infiltration(3/9),and epithelioid granulo-mas(2/9)were seen in some cases.The tumor cells expressed CD21(7/9),CD23(8/9)and CD35(9/9),partially ex-pressed SMA(6/9)and D2-40(1/9).It was noteworthy that 2 cases from the spleen accompanied by high expression of IgG4 plasma cells(80-135/10 HPF),and in the liver(0-36/10 HPF).All cases were followed up for 3-84 months,with 6 pa-tients disease-free,2 patients underwent metastasis,1 patient lost of follow-up.Conclusion EBV+IFDCS is a rare low-grade malignant tumor.EBER in situ hybridization and immunohisto-chemical detection play important roles in the diagnosis and dif-ferential diagnosis of EBV+IFDCS.Surgical resection is the main therapeutic intervention for EBV+IFDCS,and patients re-quire long-term post-surgical follow-up.

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis

Objective:To analyze the basic characteristics and change trend of mortality in HIV-infected patients aged ≥60 years in China from 2013 to 2021.Methods:The data of HIV-infected patients aged ≥60 years at diagnosis were collected from China Information System for Disease Control and Prevention to calculate the mortality density. The trajectory model was fitted using the Proc traj process in software SAS 9.4 to explore trajectory of AIDS-related mortality density and non-AIDS-related mortality density under different combinations of region, gender and age.Results:Between 2013 and 2021, a total of 244 770 HIV-infected patients were reported with 40 079 AIDS-related deaths and 50 245 non-AIDS-related deaths. The AIDS-related mortality density was 6.32 per 100 person-years, and the non-AIDS-related mortality density was 7.92 per 100 person-years, both of which showed decreasing trends over the years, and the mortality density in men was higher than that in women. Two developmental trajectories could be categorized for different trends of AIDS-related mortality density: the lower mortality density group accounted for 80.95% and showed a slow decreasing trend; the higher mortality density group accounted for 19.05% and showed a three-curve developmental trend. There were three developmental trajectories of non-AIDS-related mortality density trends: the lower mortality density group accounted for 59.52% and the medium mortality density group accounted for 28.57%, with a flat overall trend in these two groups; the higher mortality density group accounted for 11.91% with a three-curve trend.Conclusions:The mortality in HIV-infected patients aged ≥60 years in China is still high. Further attention should be paid to the early detection, diagnosis and treatment of HIV infection to effectively reduce the density of AIDS-related deaths. At the same time, attention should be paid to non-AIDS-related deaths in the elderly, and comprehensive interventions should be taken. It is necessary to conduct targeted HIV/AIDS prevention and control based on actual situation in different areas and populations

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15％ of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5％?49.7％ of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3％?17.9％, which was significantly higher than that (6.9％?7.4％) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.

Objective:To study the effects of progesterone on the morphology, proliferation, and secretion of cytokines of human decidual stromal cells (DSCs) in late pregnancy, and to explore the mechanism of progesterone in preventing spontaneous preterm birth.Methods:Human decidual stromal cells in late pregnancy were cultured and treated with different concentrations of progesterone (in the experimental groups, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L progesterone was added to the culture medium respectively, and no progesterone was added to the culture medium of control group). The morphology of DSCs was observed under the microscope, the cell length/width ratio was measured, the proliferation was detected by methyl thiazolyl tetrazolium (MTT) method, and the expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected by RT-qPCR. Results:The length/width ratios of DSCs in progesterone 10 -4 mol/L (5.87±0.19) and 10 -5 mol/L (5.98±0.27) groups were lower than that in control group (6.42±0.19), the differences were statistically significant ( P<0.001, P=0.002). The length/width ratio in the 10 -4 mol/L group was lower than that in the 10 -6 mol/L group (6.28±0.32, P=0.005). The proliferation of DSCs in the 10 -5 mol/L (0.70±0.04) and 10 -4 mol/L (0.78±0.04) groups was higher than that in control group (0.59±0.05; P=0.027, P=0.002), and proliferation of DSCs in 10 -4 mol/L group was higher than that in 10 -6 mol/L group (0.61±0.01, P=0.004). The expression of TNF-α mRNA in each progesterone group was lower than that in control group (all P<0.001) and the expression of TNF-α mRNA in the 10 -5 mol/L and 10 -4 mol/L groups was lower than that in the 10 -6 mol/L group (all P<0.001) . The expressions of IL-6 mRNA decreased gradually in control group, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L groups, the differences were statistically significant (all P<0.001). Conclusion:Progesterone can make the decidual stromal cells wider, promote proliferation, and decrease the expressions of TNF-α and IL-6 mRNA, which may play an important role in the mechanism of progesterone preventing spontaneous preterm birth.

Objective:To study the effects of progesterone on the morphology, proliferation, and secretion of cytokines of human decidual stromal cells (DSCs) in late pregnancy, and to explore the mechanism of progesterone in preventing spontaneous preterm birth.Methods:Human decidual stromal cells in late pregnancy were cultured and treated with different concentrations of progesterone (in the experimental groups, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L progesterone was added to the culture medium respectively, and no progesterone was added to the culture medium of control group). The morphology of DSCs was observed under the microscope, the cell length/width ratio was measured, the proliferation was detected by methyl thiazolyl tetrazolium (MTT) method, and the expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected by RT-qPCR. Results:The length/width ratios of DSCs in progesterone 10 -4 mol/L (5.87±0.19) and 10 -5 mol/L (5.98±0.27) groups were lower than that in control group (6.42±0.19), the differences were statistically significant ( P<0.001, P=0.002). The length/width ratio in the 10 -4 mol/L group was lower than that in the 10 -6 mol/L group (6.28±0.32, P=0.005). The proliferation of DSCs in the 10 -5 mol/L (0.70±0.04) and 10 -4 mol/L (0.78±0.04) groups was higher than that in control group (0.59±0.05; P=0.027, P=0.002), and proliferation of DSCs in 10 -4 mol/L group was higher than that in 10 -6 mol/L group (0.61±0.01, P=0.004). The expression of TNF-α mRNA in each progesterone group was lower than that in control group (all P<0.001) and the expression of TNF-α mRNA in the 10 -5 mol/L and 10 -4 mol/L groups was lower than that in the 10 -6 mol/L group (all P<0.001) . The expressions of IL-6 mRNA decreased gradually in control group, 10 -6 mol/L, 10 -5 mol/L and 10 -4 mol/L groups, the differences were statistically significant (all P<0.001). Conclusion:Progesterone can make the decidual stromal cells wider, promote proliferation, and decrease the expressions of TNF-α and IL-6 mRNA, which may play an important role in the mechanism of progesterone preventing spontaneous preterm birth.

The online version of the original article can be found at https://doi.org/10.1631/jzus.B2000190 Erratum to: J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2020 21(10):779-795 https://doi.org/10.1631/jzus.B2000190.

Objective:Analysis of the antibiotic resistance profiles and molecular typing of Salmonella typhimurium ( S. typhimurium) isolated in Wuxi city from 2011 to 2018. Methods:A total of 109 S. typhimurium isolates were detected from different types monitoring samples in Wuxi city from 2011 to 2018. Microbroth dilution method was used to test antimicrobial susceptibility of S. typhimurium for 17 antimicrobial agents. Pulsed field gel electrophoresis (PFGE) was used to conduct molecular typing. Statistical analysis was carried out using Chi-square test. Results:Tetracycline-resistance and ampicillin-resistance were most frequency in 109 S. typhimurium isolates, 69.72% (76/109) and 68.81% (75/109), respectively. For compound sulfamethoxazole, chloramphenicol, ciprofloxacin, ampicillin/sulbactam, azithromycin, nitrofurantoin, cefotaxime and aztreonam, the resistance rates were 23.85%(26/109), 22.02%(24/109), 11.93%(13/109), 4.59%(5/109), 3.67%(4/109), 3.67%(4/109), 0.92%(1/109), 0.92%(1/109), respectively. All isolates were susceptible to amikacin, ertapenem, imipenem, meropenem, ceftazidime and ceftazidime-avibactam. Azithromycin-resistance isolates were decreasing year by year gradually. The aztreonam and cefotaxime-resistant isolates were found since 2018, while chloramphenicol and compound sulfa-resistant isolates showed upward trend simultaneously. Conclusions:S. typhimurium in Wuxi city exhibited highly resistant to tetracycline and ampicillin. The significant variability existed between genotype and phenotype of S. typhimurium.