This paper presents a comprehensive study of Pulsatillae Radix,encompassing its pharmacognostic investigation,chemical constituents,and pharmacological effects.Pulsatillae Radix,first recorded in the Shen Nong's Herbal Classic,is known for its efficacy in cooling blood to stop dysentery,clearing heat,and detoxifying,and has been used to treat diseases such as enteritis caused by parasites,demonstrating potential as an antiviral candidate.Research has revealed that its major chemical constituents include triterpenoid saponins,triterpenic acids,and various other components,which collectively constitute the pharmacological foundation of Pulsatillae Radix.Significant pharmacological activities have been observed in antitumor,anti-inflammatory,antimicrobial,and antiviral aspects,providing a scientific basis for its broad application in the pharmaceutical field.

BACKGROUND: Several studies have demonstrated the occurrence of secondary tumors as a rare but significant complication of chimeric antigen receptor T (CAR-T) cell therapy, underscoring the need for a detailed investigation. Given the limited variety of secondary tumor types reported to date, a comprehensive characterization of the various secondary tumors arising after CAR-T therapy is essential to understand the associated risks and to define the role of the immune microenvironment in malignant transformation. This study aims to characterize the immune microenvironment of a newly identified secondary tumor post-CAR-T therapy, to clarify its pathogenesis and potential therapeutic targets. METHODS: In this study, the bone marrow (BM) samples were collected by aspiration from the primary and secondary tumors before and after CD19 CAR-T treatment. The CD45 + BM cells were enriched with human CD45 microbeads. The CD45 + cells were then sent for 10× genomics single-cell RNA sequencing (scRNA-seq) to identify cell populations. The Cell Ranger pipeline and CellChat were used for detailed analysis. RESULTS: In this study, a rare type of secondary chronic myelomonocytic leukemia (CMML) were reported in a patient with diffuse large B-cell lymphoma (DLBCL) who had previously received CD19 CAR-T therapy. The scRNA-seq analysis revealed increased inflammatory cytokines, chemokines, and an immunosuppressive state of monocytes/macrophages, which may impair cytotoxic activity in both T and natural killer (NK) cells in secondary CMML before treatment. In contrast, their cytotoxicity was restored in secondary CMML after treatment. CONCLUSIONS This finding delineates a previously unrecognized type of secondary tumor, CMML, after CAR-T therapy and provide a framework for defining the immune microenvironment of secondary tumor occurrence after CAR-T therapy. In addition, the results provide a rationale for targeting macrophages to improve treatment strategies for CMML treatment.
Humans ; Lymphoma, Large B-Cell, Diffuse/therapy* ; Tumor Microenvironment/genetics* ; Antigens, CD19/metabolism* ; Leukemia, Myelomonocytic, Chronic/genetics* ; Immunotherapy, Adoptive/adverse effects* ; Male ; Single-Cell Analysis/methods* ; Female ; Sequence Analysis, RNA/methods* ; Receptors, Chimeric Antigen ; Middle Aged

Objective:To explore the feasibility of joint screening of the two primary immunodeficiency diseases [severe combined immunodeficiency (SCID) and X-linked agammaglobulinemia(XLA)] and spinal muscular atrophy(SMA) in newborns by multiplex real-time quantitative PCR technology, and to provide evidence for early screening, diagnosis and treatment of children.Methods:Cross-sectional study. From July 2021 to January 2023, a total of 103 240 dry blood spots samples of newborns were collected which were delivered to Neonatal Disease Screening Center of Zhejiang by cold chain transportation. The concentrations of the T cell receptor excision ring (TREC), Kappa deletion of the recombinant excision loop (KREC), and exon 7 deletion of Survival Motor Neuron 1 (SMN1) gene in dry blood spots were simultaneously detected by multiplex real-time fluorescence quantitative PCR, taken ribonuclease P/MRP 30 000 subunits (RPP30) as an internal reference gene. The positive newborns were further diagnosed by other laboratory tests and gene sequencing was taken as gold standard. Children samples from 1 case of SCID, 3 cases of XLA and 2 cases of SMA were used for positive verification. The correlation between detected concentration of TREC/KREC and basic information in newborns were analyzed. The differences among groups for each factor were analyzed.Results:One case of SCID, 2 cases of XLA, 9 cases of SMA and 7 cases of other genetic diseases (4 cases of DiGeorge syndrome, 1 case of trisomy 21 syndrome, 1 case of Noonan syndrome and 1 case of super male syndrome) were identified by multiplex real-time fluorescence quantitative PCR. The positive predictive values of screening neonatal SCID, XLA and SMA were 2.44% (1/41), 2.78% (2/72) and 9/9 respectively. Taking the samples from clinically diagnosed 1 case of SCID, 3 cases of XLA and 2 cases of SMA as positive validation samples, which were all identified. The detected results of TREC/KREC correlated with time of blood collection, sex, weight, gestational age and delivery mode of newborns, whose r values were 0.162/0.187, 0.066/0.032, 0.045/0.042, ?0.015/?0.088 and 0.014/0.068 respectively (all P<0.05). Conclusions:Relying on current neonatal screening platform in Zhejiang, it is feasible to screen jointly two kinds of primary immunodeficiency diseases and spinal muscular atrophy in newborns by multiple real-time fluorescence quantitative PCR technology.

Objective:To investigate the value of ultra micro angiography（UMA）imaging in evaluating the inflammation activity of pediatric Crohn's disease（CD）patients.Methods:In this prospective study，71 pediatric CD patients confirmed by clinical symptoms，laboratory and ileocolonoscopical results who planed to accept infliximab injection treatments in Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from April to December 2024 were included. The CD patients were included and divided into 3 groups according to the pediatric Crohn's disease activity index（PCDAI）score：inactive CD group（ n=44），mild CD group（ n=17），and moderate-to-severe CD group（ n=10）. Before treatment，pediatric CD patients underwent B mode ultrasound（BMUS），the parameters such as bowel wall thickness（BWT），bowel wall stratification，mesenteric lymph nodes（MLNs），and existence of mesenteric fat hypertrophy（MFH）were recorded. Color Doppler flow imaging（CDFI）and UMA were used to evaluate the blood flow in the most thickened diseased bowel wall using Limberg scores. The diffierences of the above parameters among the 3 groups were compared. The diagnostic performance of UMA and CDFI in assessing the inflammatory activity of pediatric CD was evaluated. Results:There were statistically significant differences in BWT among the 3 groups（all P<0.05）. However，there were no statistically significant differences in MLNs among the 3 groups（all P>0.05）. Compared with the inactive CD，there were statistically significant differences in bowel wall stratification and MFH in both the mild and moderate-to-severe CD groups（all P<0.05）. When assessing bowel wall blood flow signals using CDFI，there was no statistically significant difference in blood flow signals in the bowel wall of the mild CD compared with the inactive CD（ P>0.05）. However，the blood flow signals in the bowel wall of the moderate-to-severe CD was significantly increased（ P<0.05）. There was also statistically significant difference in blood flow signals in the bowel wall between the mild and moderate-to-severe CD groups（ P<0.05）. When assessing bowel wall blood flow using UMA，there were statistically significant differences in blood flow signals in the bowel wall among all 3 groups（all P<0.05）. As the degree of inflammation increased，the blood flow signals in the bowel wall also increased significantly. ROC analysis revealed that diagnostic performance of the UMA-BMUS model was significantly better than that of the CDFI-BMUS model（AUC 0.945 vs. 0.873，sensitivity 88.9% vs. 74.1%，specificity 97.7% vs. 95.5%；all P<0.05）. Conclusions:By sensitively depicting the low-velocity and tiny blood flow signals of the inflamed bowel wall，UMA is an effective noninvasive imaging method to make semi-quantitative evaluation of the activity of pediatric Crohn's disease.

Objective:To evaluate the efficacy and safety of transdermal delivery of compound glycyrrhizin injection as an adjunctive treatment for erythematotelangiectatic rosacea (ETR).Methods:This was a randomized controlled trial conducted from March to October 2024. At Sichuan Provincial People′s Hospital, 60 patients with newly diagnosed ETR were prospectively enrolled and randomized by a random number table into study group [ n=30; 6 male and 24 female; aged 18-60 (38.9±9.8) years] and control group [ n=30; 4 male and 26 female; aged 18-60 (35.7±10.1) years]. The study group received transdermal delivery of compound glycyrrhizin injection by a medium-frequency drug-delivery therapeutic apparatus together with oral azithromycin and hydroxychloroquine sulfate, whereas the control group received oral azithromycin and hydroxychloroquine sulfate. The efficacy evaluations were conducted at baseline and at weeks 2, 4, 6, and 8 post-treatment. Outcome measures included percentage of erythema area, stratum corneum hydration, transepidermal water loss (TEWL), clinician′s erythema assessment (CEA), erythema and telangiectasia scores, dermatology life quality index (DLQI), and efficacy rate. Adverse reactions during treatment were also recorded. Results:Compared with baseline, both groups exhibited significant reductions in percentage of erythema area, TEWL, CEA, erythema and telangiectasia score, and DLQI, and significant increases in stratum corneum hydration at each post-treatment time point (all P<0.05). After 2, 4, 6, and 8 weeks of treatment, the percentage of erythema area, erythema and telangiectasia scores in the study group were all lower than those in the control group, while the stratum corneum hydration level in the study group was higher than that in the control group (all P<0.05). After 4, 6, and 8 weeks of treatment, TEWL, CEA, and DLQI in the study group were all lower than those in the control group (all P<0.05). After 4 weeks of treatment, the efficacy rate in the study group was 56.7% (17/30), which was higher than that of the control group at 23.3% (7/30, P=0.046). After 6 weeks of treatment, the efficacy rate in the study group was 83.3% (25/30), higher than that of the control group at 50.0% (15/30, P=0.020). After 8 weeks of treatment, the efficacy rate in the study group was 86.7% (26/30), higher than that of the control group at 66.7% (20/30, P<0.001). No severe adverse reactions were observed in either group. Conclusion:Transdermal delivery of compound glycyrrhizin injection as an adjunctive treatment for ETR demonstrates favorable efficacy and good safety.

Objective:To analyze the composition and diversity characteristics of facial microbial communities in patients with moderate acne.Methods:This prospective study enrolled 30 patients with moderate acne [12 males, 18 females; aged 21-30 (25.4±2.5) years] from the Department of Dermatology, Sichuan Provincial People′s Hospital from March to July 2021. Thirty healthy controls [13 males, 17 females; aged 24-29 (25.2±1.4) years] were included during the same period. Facial skin swabs were collected from both groups. Total DNA was extracted, followed by PCR amplification, library preparation, and PE250 sequencing. After splicing, filtering, denoising, and chimera removal, amplicon sequence variants (ASV) feature tables and representative sequences were generated to compare microbial community differences between the two groups.Results:A total of 60 samples were sequenced, yielding 2 021 342 valid sequences. The 16S rRNA gene sequences were clustered into 8 379 ASV, with 589 ASV shared between the two groups, while 6 445 ASV were uniquely identified in healthy controls. At the phylum level, both groups showed similar dominant phyla: Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. At the genus level, the acne group was predominantly colonized by Ralstonia (relative abundance 31.85%) and Staphylococcus (28.04%), while healthy controls exhibited more balanced distributions, primarily Staphylococcus (9.18%) and Enhydrobacter (7.37%). Alpha diversity analysis, Beta diversity analysis, and LefSe analysis revealed statistically significant differences in microbial communities between groups ( R2=0.157, P<0.001). The acne group showed lower microbial richness and evenness compared to healthy controls (both P<0.001). Conclusion:Patients with moderate acne exhibit microecological imbalance in facial microbial communities, characterized by reduced microbial richness and evenness.

Objective:To investigate the efficacy and safety of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for the treatment of BCR::ABL-negative chronic neutrophilic leukemia (CNL) and MDS/MPN with neutrophilia.Methods:This study retrospectively analyzed 12 cases of CNL and MDS/MPN with neutrophilia that underwent allo-HSCT from March 2017 to June 2024, comprising 7 males and 5 females with a median age of 48 ( IQR: 28, 59) years. The 2-year overall survival (OS), disease-free survival (DFS), cumulative incidence of relapse (CIR), and transplantation-related mortality (TRM) rates were analyzed. Complications were also assessed. Results:Of the 12 patients, 6 received matched sibling HSCT and 6 received haploidentical HSCT. All patients had successful engraftment, and the median times of neutrophil and platelet engraftment were 17 ( IQR: 11, 24) days and 15 ( IQR: 9, 28) days, respectively. Grade Ⅱ–Ⅳ acute graft versus host disease (GVHD) and chronic GVHD occurred in 2 and 4 cases, respectively. The 2-year OS, DFS, CIR, and TRM rates were (65.6 ± 16.4) %, (41.7 ± 16.6) %, (47.2 ±18.2) %, and (11.1 ± 11.4) %, respectively, after a median follow-up time of 637 ( IQR: 330, 943) days. One patient died from treatment-related complications due to respiratory failure caused by coronavirus disease 2019. Two patients died due to relapse. Conclusion:Allo-HSCT can be applied as a safe and effective approach to treat CNL and MDS/MPN with neutrophilia.

Objective To investigate the bacterial contamination of water specimens at different pre-discharge time of flexible endoscopic final rinse water before daily use,and provide reference basis for choosing the optimal pre-dis-charge time in clinical work.Methods From August to December 2023,the water produced by the purified water equipment of the digestive endoscopy centers of 4 tertiary first-class medical institutions in Jiangxi Province,as well as the final rinsing water at the terminal-point at different pre-discharge times(before pre-discharge,discharge of 1,3,5,and 10 minutes)were subjected to on-site sampling,the collected water specimens were performed microbio-logical cultures by pouring and filter membrane methods,unqualified water specimens were performed bacterial i-dentification.Results 48 specimens of water produced by purified water equipment were collected,with qualified rate of 100％.480 final water specimens were collected,with a qualified rate of 0 before pre-discharge.The quali-fied rates of pre-discharge at 1,3,5,and 10 minutes were 0,21.88％,55.21％,and 73.96％,respectively.Bac-terial colony counts in the final rinse water with different pre-discharge times in 4 medical institutions showed statis-tically significant differences(all P＜0.001).As the pre-discharge time increased,bacterial colony counts in the fi-nal rinse water of all four medical institutions showed a decreasing trend.The pairwise comparison of the overall bacterial colony count of the final rinse water at different pre-discharge time showed that there was no statistically significant difference in the bacterial colony count of the final rinse water at 5 and 10 minutes of pre-discharge(P＞0.05),while the pairwise comparison between other time points showed statistically significant differences(all P＜0.001).Environmental contaminated bacteria such as Micrococcus luteus,Cupriavidus paucula,and Sphin-gononas paucimobilis were detected from the unqualified water specimens.Conclusion With the extension of pre-discharge time,the degree of bacterial contamination in the final rinse water significantly decreases.All levels of medical institutions need to pre-discharge the final rinse water daily,and the pre-discharge time should last for at least 5 minutes.

Diagnosis-related groups (DRG) payment is an important component of deepening the reform of medical insurance payment methods. In June 2023, a tertiary hospital launched an intelligent DRG grouping audit system to enhance grouping accuracy. By establishing a multi departmental collaborative organizational structure, building a standardized knowledge base and a rule base covering five categories (diagnosis, fees, testing, nursing, and pathology), and integrating electronic medical records, medical orders, testing, and imaging data throughout the entire diagnosis and treatment process, the intelligent DRG grouping audit system with data collection, identification, extraction, comparison, and output modules was constructed to achieve intelligent audit. At the same time, it was formed a closed-loop management system for pre reporting quality control, in-process group entry control, and post data analysis and assessment, which would prevent the risk of differentiated behaviors such as high coding and high sets, and ensure the reasonable use of medical insurance funds. By January 2024, the system had covered 89 ADRG groups, and improved the efficiency and quality of DRG grouping audit. Compared with February to May 2023, the monthly average rejection rate of medical records on the first page decreased by 9.4% after the system was put into operation (June to December 2023), and core medical indicators such as the number of DRG groups, medical insurance settlement cases, and time consumption index continued to improve. The practical experience could provide reference and inspiration for other hospitals in China.

Objective:To develop an artificial intelligence (AI)-based platelet review strategy to identify abnormal platelet histograms with no significant difference between initial impedance platelet count (PLT-I) and PLT-F results.Methods:This study included 5 119 routine blood analysis in Nanfang Hospital of Southern Medical University and its Ganzhou branch from July 2023 and March 2024. Specimens exhibiting abnormal platelet histograms and an initial platelet count >40×10?/L underwent review using the fluorescent platelet count (PLT-F) channel. Consistency of the results was defined as a difference between impedance platelet count (PLT-I) and PLT-F less than ±20% of the PLT-F results. A deep learning model was developed using platelet and red blood cell histogram data from a training set of 3 807 specimens. The model′s diagnostic performance was evaluated on an independent external validation set ( n=805) using receiver operating characteristic (ROC) curve analysis. Changes in the number of reviewed samples and sample turnaround time were analyzed to assess its clinical utility. Results:The deep learning model based on platelet and red blood cell histograms achieved an area under the ROC curve (AUC) of 0.854 in the training set. At a cutoff value of 0.1, the sensitivity was 0.954 and specificity was 0.358. The model could reduce review by 16.80% (190/1 131). In the validation set, the AUC was 0.805, with a sensitivity of 0.955 and specificity of 0.307, corresponding to a reduction of 17.41% (47/270) in reviewed specimens.Conclusion:The platelet review prediction model developed based on deep learning technology can efficiently identify samples with consistent results before and after review, reducing unnecessary reviews and shortening specimen testing time, thereby improving the efficiency of platelet test.