ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps （NETs） and oxygen-glucose deprivation （OGD）. MethodsFirst， dimethyl sulfoxide （DMSO） was used to induce the differentiation of the human promyelocytic leukemia cell line （HL-60） into neutrophil-like cells （dHL-60）， and cell differentiation was assessed through Giemsa staining and flow cytometry. Next， phorbol myristate （PMA） was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining， and NETs were extracted by low-speed centrifugation. Finally， the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group， a model （NETs + OGD） group， a traditional Chinese medicine （NETs + OGD + 5% Huayu Jiedu prescription-containing serum） group， and a NETosis inhibitor （100 mg·L^-1 DNase I + NETs + OGD） group. The cell counting kit-8 （CCK-8） assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum， as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the levels of myeloperoxidase-DNA complexes （MPO-DNA）， citrullinated histone H3 （CitH3）， and neutrophil elastase （NE）. Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay （TUNEL） staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group， cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli， distinct reniform and lobulated changes of nuclei， highly condensed chromatin， and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity （MFI） of CD11b in the model group was enhanced （P<0.01）. Compared with those in the normal control group， dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity， with chromatin released from the cytoplasm to form abundant extracellular， cloud-like fibrillar structures， and a marked increase in fluorescence intensity （P<0.01）. Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells， the optimal concentration for NETs was determined as 1 000 ng·L^-1， the optimal concentration of Huayu Jiedu formula-containing serum as 5%， and the optimal duration for OGD as 2 h. Compared with the normal control group， the model group showed elevated levels of MPO-DNA， CitH3， and NE （P<0.01） and upregulated protein levels of CitH3 and cleaved Caspase-3 （P<0.01）. Furthermore， TUNEL staining revealed an increased proportion of apoptotic cells （P<0.01） and flow cytometry showed an increased apoptosis rate （P<0.01） in the model group. Compared with the model group， the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA， CitH3， and NE （P<0.05，P<0.01） and downregulated protein levels of CitH3 and cleaved Caspase-3 （P<0.01）. In addition， TUNEL staining showed a decrease in the proportion of apoptotic cells （P<0.01）， and flow cytometry revealed a decrease in the apoptosis rate （P<0.01）. ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity

ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps （NETs） and oxygen-glucose deprivation （OGD）. MethodsFirst， dimethyl sulfoxide （DMSO） was used to induce the differentiation of the human promyelocytic leukemia cell line （HL-60） into neutrophil-like cells （dHL-60）， and cell differentiation was assessed through Giemsa staining and flow cytometry. Next， phorbol myristate （PMA） was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining， and NETs were extracted by low-speed centrifugation. Finally， the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group， a model （NETs + OGD） group， a traditional Chinese medicine （NETs + OGD + 5% Huayu Jiedu prescription-containing serum） group， and a NETosis inhibitor （100 mg·L^-1 DNase I + NETs + OGD） group. The cell counting kit-8 （CCK-8） assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum， as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the levels of myeloperoxidase-DNA complexes （MPO-DNA）， citrullinated histone H3 （CitH3）， and neutrophil elastase （NE）. Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay （TUNEL） staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group， cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli， distinct reniform and lobulated changes of nuclei， highly condensed chromatin， and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity （MFI） of CD11b in the model group was enhanced （P<0.01）. Compared with those in the normal control group， dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity， with chromatin released from the cytoplasm to form abundant extracellular， cloud-like fibrillar structures， and a marked increase in fluorescence intensity （P<0.01）. Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells， the optimal concentration for NETs was determined as 1 000 ng·L^-1， the optimal concentration of Huayu Jiedu formula-containing serum as 5%， and the optimal duration for OGD as 2 h. Compared with the normal control group， the model group showed elevated levels of MPO-DNA， CitH3， and NE （P<0.01） and upregulated protein levels of CitH3 and cleaved Caspase-3 （P<0.01）. Furthermore， TUNEL staining revealed an increased proportion of apoptotic cells （P<0.01） and flow cytometry showed an increased apoptosis rate （P<0.01） in the model group. Compared with the model group， the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA， CitH3， and NE （P<0.05，P<0.01） and downregulated protein levels of CitH3 and cleaved Caspase-3 （P<0.01）. In addition， TUNEL staining showed a decrease in the proportion of apoptotic cells （P<0.01）， and flow cytometry revealed a decrease in the apoptosis rate （P<0.01）. ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity

Objective To investigate the perioperative use of antimicrobial agents in class Ⅰ incisional ophthalmic surgeries in secondary first-class and above medical institutions in Shanghai City,and provide a basis for the rational use of antimicrobial agents in such surgeries.Methods Perioperative antimicrobial prophylaxis(PAP)in patients undergoing class Ⅰ incisional ophthalmic surgery reported by secondary first-class and above hospitals in Shanghai City from 2017 to 2022 was collected retrospectively.The prophylactic use rate,types,rationality of selection,and timing of use of antimicrobial agents were analyzed comprehensively.Results From 2017 to 2022,a total of 54 868 cases of class Ⅰ incisional ophthalmic surgeries were reported from secondary first-class and above hospitals in Shanghai City.The PAP rates in tertiary comprehensive hospitals,tertiary specialty hospitals,tertiary traditional Chinese medicine hospitals,and secondary comprehensive hospitals were 4.72％,1.79％,3.22％,and 6.63％,re-spectively.The overall PAP rate showed a yearly decreasing trend,from 6.39％in 2017 to 2.31％in 2021.Among different types of hospitals,the PAP rate in secondary comprehensive hospitals decreased most significantly,from 12.72％in 2017 to 0.53％in 2022.The main types of prophylactic antimicrobial use were first-,second-and third-generation cephalosporins,and quinolones.The rational selection rates of antimicrobial agents were highest in ter-tiary comprehensive hospitals(17.13％)and lowest in tertiary specialty hospitals(1.08％).The PAP rates in sys-temic veins 0.5-1 hour before surgery were highest in tertiary specialty hospitals(88.17％)and lowest in tertiary comprehensive hospitals(71.53％).The withdrawal rates of antimicrobial agents within 24 hours after surgery were highest in tertiary traditional Chinese medicine hospitals(80.87％)and lowest in tertiary specialty hospitals(13.26％).Conclusion The PAP for class Ⅰ incisional ophthalmic surgeries in secondary first-class and above medical institutions in Shanghai City still presents certain irrational phenomena in terms of antimicrobial types and administration routes.The administration rate 0.5-1 hour before surgery and withdrawal rate of antimicrobial agents within 24 hours after surgery need to be further improved.Enhanced management is necessary to promote more rational use of antimicrobial agents.

Objective:To compare the performance of next-generation sequencing (NGS) and Sanger sequencing in investigating somatic hypermutation (SHM) status of immunoglobulin heavy chain variable region (IGHV) genes. It specifically focuses on identifying key factors contributing to discrepancies between the two methods, particularly under complex clonal backgrounds, to inform optimized strategies for clinical application.Methods:This retrospective analysis included 53 samples, comprising 43 identified as non-monoclonal and 10 as monoclonal using Sanger sequencing. All samples were further analyzed using NGS to assess IGHV SHM. The two methods were used for systematic comparison. For discordant cases, in-depth attribution analysis was conducted, considering factors, including clonal abundance quantification, differences in primer design, and interpretation criteria.Results:Among the 53 patients who underwent both Sanger and NGS testing, 36 were male and 17 were female, with a median age of 64 years (range: 33–88). Diagnoses included chronic lymphocytic leukemia (CLL) in 35 (66.0% ), diffuse large B-cell lymphoma in 9 (17.0% ), follicular lymphoma in 3 (5.7% ), mantle cell lymphoma in 3 (5.7% ), and other types in 3 (5.7% ) cases. In the 43 cases with non-monoclonal profiles using Sanger sequencing, NGS revealed 23 cases as biclonal or polyclonal, 17 as monoclonal, and 3 with no detectable clonality. The primary discrepancies between the two methods involved variations in clonality assessment, IGHV gene rearrangement types, and mutation rates. Among the 10 cases identified as monoclonal using Sanger sequencing, NGS detected biclonality and markedly different IGHV rearrangement types in 2 and 4 cases, respectively. Minor differences were observed in SHM percentage between the two methods; however, these did not substantially affect the overall determination of mutational status.Conclusion:Compared with Sanger sequencing, NGS exhibits superior performance in assessing IGHV SHM status under complex clonal conditions. It provides greater sensitivity and accuracy in detecting subclonal components and quantifying clonal proportions, thereby providing a more precise molecular basis for diagnosing and prognostically assessing lymphoid malignancies, including CLL.

Objective To investigate the perioperative use of antimicrobial agents in class Ⅰ incisional ophthalmic surgeries in secondary first-class and above medical institutions in Shanghai City,and provide a basis for the rational use of antimicrobial agents in such surgeries.Methods Perioperative antimicrobial prophylaxis(PAP)in patients undergoing class Ⅰ incisional ophthalmic surgery reported by secondary first-class and above hospitals in Shanghai City from 2017 to 2022 was collected retrospectively.The prophylactic use rate,types,rationality of selection,and timing of use of antimicrobial agents were analyzed comprehensively.Results From 2017 to 2022,a total of 54 868 cases of class Ⅰ incisional ophthalmic surgeries were reported from secondary first-class and above hospitals in Shanghai City.The PAP rates in tertiary comprehensive hospitals,tertiary specialty hospitals,tertiary traditional Chinese medicine hospitals,and secondary comprehensive hospitals were 4.72％,1.79％,3.22％,and 6.63％,re-spectively.The overall PAP rate showed a yearly decreasing trend,from 6.39％in 2017 to 2.31％in 2021.Among different types of hospitals,the PAP rate in secondary comprehensive hospitals decreased most significantly,from 12.72％in 2017 to 0.53％in 2022.The main types of prophylactic antimicrobial use were first-,second-and third-generation cephalosporins,and quinolones.The rational selection rates of antimicrobial agents were highest in ter-tiary comprehensive hospitals(17.13％)and lowest in tertiary specialty hospitals(1.08％).The PAP rates in sys-temic veins 0.5-1 hour before surgery were highest in tertiary specialty hospitals(88.17％)and lowest in tertiary comprehensive hospitals(71.53％).The withdrawal rates of antimicrobial agents within 24 hours after surgery were highest in tertiary traditional Chinese medicine hospitals(80.87％)and lowest in tertiary specialty hospitals(13.26％).Conclusion The PAP for class Ⅰ incisional ophthalmic surgeries in secondary first-class and above medical institutions in Shanghai City still presents certain irrational phenomena in terms of antimicrobial types and administration routes.The administration rate 0.5-1 hour before surgery and withdrawal rate of antimicrobial agents within 24 hours after surgery need to be further improved.Enhanced management is necessary to promote more rational use of antimicrobial agents.

Objective:To compare the performance of next-generation sequencing (NGS) and Sanger sequencing in investigating somatic hypermutation (SHM) status of immunoglobulin heavy chain variable region (IGHV) genes. It specifically focuses on identifying key factors contributing to discrepancies between the two methods, particularly under complex clonal backgrounds, to inform optimized strategies for clinical application.Methods:This retrospective analysis included 53 samples, comprising 43 identified as non-monoclonal and 10 as monoclonal using Sanger sequencing. All samples were further analyzed using NGS to assess IGHV SHM. The two methods were used for systematic comparison. For discordant cases, in-depth attribution analysis was conducted, considering factors, including clonal abundance quantification, differences in primer design, and interpretation criteria.Results:Among the 53 patients who underwent both Sanger and NGS testing, 36 were male and 17 were female, with a median age of 64 years (range: 33–88). Diagnoses included chronic lymphocytic leukemia (CLL) in 35 (66.0% ), diffuse large B-cell lymphoma in 9 (17.0% ), follicular lymphoma in 3 (5.7% ), mantle cell lymphoma in 3 (5.7% ), and other types in 3 (5.7% ) cases. In the 43 cases with non-monoclonal profiles using Sanger sequencing, NGS revealed 23 cases as biclonal or polyclonal, 17 as monoclonal, and 3 with no detectable clonality. The primary discrepancies between the two methods involved variations in clonality assessment, IGHV gene rearrangement types, and mutation rates. Among the 10 cases identified as monoclonal using Sanger sequencing, NGS detected biclonality and markedly different IGHV rearrangement types in 2 and 4 cases, respectively. Minor differences were observed in SHM percentage between the two methods; however, these did not substantially affect the overall determination of mutational status.Conclusion:Compared with Sanger sequencing, NGS exhibits superior performance in assessing IGHV SHM status under complex clonal conditions. It provides greater sensitivity and accuracy in detecting subclonal components and quantifying clonal proportions, thereby providing a more precise molecular basis for diagnosing and prognostically assessing lymphoid malignancies, including CLL.

ObjectiveTo compare the air disinfection effects of different human-machine coexistence disinfection methods in the high-risk areas of airborne diseases in medical and healthcare institutions, and to provide a reference for the prevention and control of airborne diseases in medical and healthcare institutions. MethodsField trials were conducted in the fever clinic, the infection disease department, and dental clinics of a tertiary hospital in Shanghai, respectively. The existing air disinfection methods (plasma air disinfection machine, circulating air ultraviolet disinfection machine or negative pressure ventilation system), upper-room 222 nm ultraviolet germicidal system, and the combination of the existing air disinfection methods and upper-room 222 nm ultraviolet germicidal system were all used in each location in the experiment group. The control group did not adopt specific air disinfection methods. Air sampling was conducted by the six-level sieve hole microbial sampler or the flat slab exposure method. The daily air sampling time was from 8:00 a.m. to 16:00 p.m., with one sample per hour, and a total of 9 samples were taken. The disinfection effects were compared by calculating the total number of airborne bacteria colonies and the sterilization rate for each disinfection method. ResultsThe total numbers of airborne bacteria colonies in the fever outpatient infusion room, the ward and nurse station of infection disease department of 222 nm group were lower than that in the control group (P=0.005, P<0.001, P<0.001). The total numbers of airborne bacteria colonies in the fever outpatient infusion room and the dental examination room of 222 nm group were lower than that in the control group or plasma air disinfection machine group (P=0.022, P=0.014). The total numbers of airborne bacteria colonies in the nucleic acid sampling room of plasma air disinfection machine group combined with 222 nm group were lower than that in plasma air disinfection machine group (P=0.019). The total numbers of airborne bacteria colonies in the CT examination room of fever clinic of the 222 nm group were lower than that in the circulating air ultraviolet disinfection machine group (P=0.002). The total numbers of airborne bacteria colonies in the CT examination room of 222 nm group combined with circulating air ultraviolet disinfection machine were lower than that of circulating air ultraviolet disinfection machine group and the control group (P=0.008, P<0.001). The air sterilization rate of upper-room 222 nm ultraviolet germicidal system ranged from 48.04% to 73.74%. The air sterilization rate of plasma air/circulating air ultraviolet disinfection machine combined with the upper-room 222 nm ultraviolet germicidal system ranged from 6.86% to 73.77%. ConclusionUpper-room 222 nm ultraviolet germicidal system could effectively reduce airborne colonies in the air and improve air hygiene quality in both clinic and ward environments with high airborne transmission risks.

The seat of human intelligence is the human cerebral cortex, which is responsible for our exceptional cognitive abilities. Identifying principles that lead to the development of the large-sized human cerebral cortex will shed light on what makes the human brain and species so special. The remarkable increase in the number of human cortical pyramidal neurons and the size of the human cerebral cortex is mainly because human cortical radial glial cells, primary neural stem cells in the cortex, generate cortical pyramidal neurons for more than 130 days, whereas the same process takes only about 7 days in mice. The molecular mechanisms underlying this difference are largely unknown. Here, we found that bone morphogenic protein 7 (BMP7) is expressed by increasing the number of cortical radial glial cells during mammalian evolution (mouse, ferret, monkey, and human). BMP7 expression in cortical radial glial cells promotes neurogenesis, inhibits gliogenesis, and thereby increases the length of the neurogenic period, whereas Sonic Hedgehog (SHH) signaling promotes cortical gliogenesis. We demonstrate that BMP7 signaling and SHH signaling mutually inhibit each other through regulation of GLI3 repressor formation. We propose that BMP7 drives the evolutionary expansion of the mammalian cortex by increasing the length of the neurogenic period.
Animals ; Mice ; Humans ; Ependymoglial Cells/metabolism* ; Hedgehog Proteins/metabolism* ; Ferrets/metabolism* ; Cerebral Cortex ; Neurogenesis ; Mammals/metabolism* ; Neuroglia/metabolism* ; Bone Morphogenetic Protein 7/metabolism*

Purpose To assess the feasibility of nuclear score combined with ultrasound features on indeterminate thyroid nodules in fine needle aspiration cytological diagnosis.Methods A consecutive cohort of 119 thyroid fine-needle aspiration(FNA)specimens with indeterminate diagnosis(TBSRTC cate-gory Ⅲ-Ⅴ)and available histopathologic follow-up was evalua-ted.Original sonographic images and cytological slides of each nodule were reviewed by two experienced sonographers and two experienced cytopathologists respectively,and evaluated jointly to obtain consensus.The identification of the optimal cut-off points of simplified nuclear score and ultrasound features score for the diagnosis of malignancy or low-risk neoplasm were evalu-ated using the receiver operating characteristic(ROC)curves and the assessment of the area under the ROC curve(AUC).The specificity,sensitivity,positive predictive value(PPV)and negative predictive value(NPV)of nuclear score and US score were evaluated from crosstabs based on Cut-off points and signif-icance were calculated.The diagnostic accuracy of simplified nuclear score combined with ultrasound features was estimated by Logistic regression and ROC curve analysis.Results Nucle-ar grooves,intra-nuclear inclusions and chromatin clearing were more common in malignancy/low-risk neoplasms(P = 0.001,0.013 and 0.001 respectively).A Cut-off point of≥2 for the simplified nuclear score was sensitive for defining malignancy/low-risk neoplasm,and the sensitivity was 99.1%.Nodule height greater than width,absent halo sign,irregular margin and absent intranodular vascularization were more common in malig-nancy/low-risk neoplasms(P = 0.003,0.007,0.003 and 0.004 respectively).When the Cut-off point was set as≥5,the PPV and specificity for defining malignancy/low-risk neoplasm were 95.9%and 78.6%respectively.ROC curve analysis showed that the diagnostic accuracy of nuclear score combined with US score(0.873,95%CI = 0.777-0.970)was higher than that of nuclear score alone(0.817,95%CI = 0.687-0.948,P =0.092)or US score alone(0.758,95%CI 0.637-0.879,P =0.047).But no significant difference was detec-ted between the combined score and the nuclear score.Conclu-sion The addition of simplified nuclear score and US score to FNA cytology can increase the diagnostic accuracy of cytology in indeterminate categories.

Objective:To investigate the dynamic change and clinical impact of DEK-NUP214 fusion gene in patients with acute myeloid leukemia (AML) receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Real-time quantitative polymerase chain reaction (RQ-PCR) and multicolor flow cytometry (FCM) were used to detect DEK-NUP214 gene expression and leukemia-associated immunophenotype (LAIP) in 15 newly diagnosed patients with positive DEK-NUP214 and receiving allo-HSCT from September 2012 to September 2017 at Peking University People′s Hospital. The clinical outcome was analyzed using Kaplan-Meier survival curves. The impact of DEK-NUP214 expression was analyzed by log-rank test.Results:The subjects were followed-up with a median period of 657 (62-2 212) days. The median DEK-NUP214 expression level at diagnosis was 488% (274%-1 692%). Thirteen patients achieved complete remission before allo-HSCT. Thirteen patients had a residual DEK-NUP214 expression of 0.38% (0.029%-738.9%) before allo-HSCT. After allo-HSCT, DEK-NUP214 expression in 9/13 patients remained positive, which dropped by around 500 folds (5.7-5 663.0 folds) within a month post-transplant. Five patients died and 2 patients relapsed. The 3-year cumulative incidence of relapse in patients with positive DEK-NUP214 before transplant was 17.5%±11.3% and the 3-year overall survival was 60.5%±13.8%. After allo-HSCT, DEK-NUP214-negative patients had a better outcome.Conclusion:Quantitative monitor of DEK-NUP214 fusion gene could be a sensitive indicator of MRD status after allo-HSCT.