Objective: To investigate the correlation between color parameters (L^*, a^*, b^*, Eab^*) and the contents of reducing sugars, total polysaccharides, total oligosaccharides, as well as four saccharides (fructose, glucose, sucrose, and kestose) during the nine cycles of steaming and sun-drying processing of Polygonatum kingianum, and to preliminarily explore the optimal processing duration.
Methods: The color changes were objectively evaluated using a colorimeter. The anthrone-sulfuric acid method was employed to determine total polysaccharides and oligosaccharides. The 3,5-dinitrosalicylic acid (DNS) colorimetric method was used to measure total reducing sugar content. High-performance liquid chromatography coupled with charged aerosol detection (HPLC-CAD) was applied for quantitative analysis of fructose, glucose, sucrose, and kestose. Multivariate statistical analysis was conducted to assess samples from different processing stages.
Results: Significant variations in color and component contents were observed across processing stages. The herbal pieces progressively darkened with increased processing cycles: brightness (L^*) and total color difference (Eab^*) initially decreased then stabilized, while a^* (red-green) and b^* (yellow-blue) values first increased then declined. Total polysaccharides and oligosaccharides showed overall decreasing trends, whereas reducing sugars initially increased before stabilizing. Fructose and glucose levels rose continuously, while sucrose and kestose decreased progressively, becoming undetectable after five cycles.
Conclusion: The chromatic alteration and saccharide composition of P. kingianum showed significant correlation with processing duration. Both total color difference (Eab^*) and reducing sugar content stabilized after four processing cycles (12 hours), suggesting that four cycles of steaming and sun-drying may represent the optimal processing duration.

Objective: To investigate the correlation between the color parameters (L^*, a^*, b^*, Eab^* values) of Polygoni Multiflori Radix Praeparata powder prepared by different processing techniques and the contents of 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucopyranoside, emodin, physcion, emodin-8-O-β-D-glucopyranoside, physcion-8-O-β-D-glucopyranoside.
Methods: The L^*, a^*, b^* and Eab^* values of Polygoni Multiflori Radix Praeparata powder prepared by different processing techniques were determined by spectrophotometer, and the contents of the five components were determined by high performance liquid chromatography. Secondly, SPSS 26.0 software and Simca 14.1 software were used to analyze the correlation.
Results: Through the hierarchical cluster analysis (HCA), it was found that the steamed samples and black bean steamed samples could be obviously divided into two categories: raw products and processed products. The processed products could be further divided into 2-8 h and 12-48 h. Pearson correlation analysis showed that the content of stilbene glycoside was significantly positively correlated with L^*, a^* and b^* values (P<0.01). The a^* value was significantly positively correlated with the content of emodin and physcion (P<0.01). Emodin-8-O-β-D-glucoside was positively correlated with L^* value and b^* value, while physcion-8-O-β-D-glucoside was negatively correlated with a^* value. Partial least squares discriminant analysis (PLS) showed that 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucoside (VIP=1.69) and emodin-8-O-β-D-glucoside (VIP=1.06) were the key variables affecting chromaticity characteristics (P<0.01).
Conclusion: The three processes of steaming, black bean steaming and black bean stewing are consistent in composition transformation and chromaticity variation, and stilbene glycoside can be used as a specific index component to characterize the processed color. Chromatic parameters can effectively reflect the processing progression and serve as quality monitoring indicators during production.

Objective To investigate the susceptibility and infection characteristics of dengue virus(DENV)in cells derived from diverse tissues of tree shrews and to provide a basis for expanding the repertoire of DENV-permissive cell models in this species.Methods DENV was inoculated at a multiplicity of infection(MOI)of 0.02 into tree shrew skin fibroblasts(TSFs),primary tree shrew renal epithelial cells(pTRECs),tree shrew aortic endothelial cells(TAECs),tree shrew aortic smooth muscle cells(TASMCs),tree shrew hepatocytes(THs),tree shrew corneal stromal cells(TCSCs),tree shrew brain microvascular endothelial cells(TBMECs),and tree shrew retinal microvascular endothelial cells(TRMECs).C6/36,Vero,A549,and BHK-21 cells(commonly used for DENV propagation)were used as positive controls.Over 6 days post-infection,cellular cytopathic effects were monitored at 12-hour intervals using an inverted microscope,viral RNA loads in cell lysates were quantified by real-time fluorescent quantitative PCR to generate proliferation curves,and viral titers were determined by plaque assay.Results Seven types of tree shrew cells,except TRMECs,were susceptible to DENV.Prolonged infection induced pronounced cytopathic effects,including cell rounding,detachment,necrosis,and lysis,across all susceptible cells.The viral RNA loads detected in lysates of pTRECs,TBMECs,TASMCs,TAECs and THs,approached those of positive controls(≥4×107 copies/μL).Infectious progeny viruses were produced by these five cell types,with three(TAECs,3.13×105 PFU/mL;THs,2.03×105 PFU/mL;pTRECs,1.58×105 PFU/mL)exhibiting titers comparable to C6/36(3.85×10 5 PFU/mL)and earlier viral harvests.Conclusion DENV exhibits broad susceptibility to tree shrew cells of multiple tissue origins,with proliferation rates surpassing those of conventional cell lines sourced from other species.TAECs,THs,and pTRECs are particularly suitable for large-scale DENV proliferation,suggesting their potential involvement in in vivo infection.

Objective:To explore the effect of bone morphogenetic with protein 2 (BMP2) intervention in iron metabolism related protein expression on rats osteoporosis (OP) by regulating hepcidin level.Methods:Thirty-six 8-week-old SPF grade female SD rats with a body weight of 200-220 g were divided into sham group (Removal of adipose tissue around ovaries), model group (ovariectomy was used to prepare postmenopausal OP model rats), adenovirus group (OP model rats injected with adenovirus into the femoral cavity), and overexpression BMP2 adenovirus group (OP model rats injected with BMP2 overexpressing adenovirus into the femoral cavity) using a random number table method, with 9 rats in each group. Rats were euthanized after 2 months of injection, and femoral tissue was taken for micro computed tomography (micro-CT) three-dimensional reconstruction to observe the microstructural characteristics of bone trabeculae, and calculate bone mineral density (BMD), trabecular number (Tb.N), trabecular thickness (Tb.Th), bone volume/total volume (BV/TV), and trabecular separation (Tb.Sp) values. Hematoxylin eosin staining, Masson staining, and tartrate-resistant acid phosphatase (TRAP) staining were used to observe the pathological damage, collagen content, and osteoclast count of femoral tissues in four group rats. Malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), ferritin, hepcidin levels in serum were detected. Real-time quantitative PCR was used to detect BMP2 mRNA expression in femoral tissue. Western blotting were used to detect BMP2, ALP, OCN, OPN, RANKL, TRAP, CTX-I, hepcidin, FPN1, TfR1, DMT1 protein expression in femoral tissue.Results:The differences of BMD, Tb.N, Tb.Th, BV/TV, and Tb.Sp in sham group, model group, adenovirus group, and BMP2 overexpressing adenovirus group were statistically significant ( P<0.05), among them, BMD, Tb.N, Tb.Th, and BV/TV in model group were lower than those in sham group, Tb.Sp was higher than that in sham group, BMD, Tb.N, Tb.Th, and BV/TV in BMP2 overexpressing adenovirus group were higher than those in model group and adenovirus group, Tb.Sp was lower than that in model group and adenovirus group ( P<0.05). Sham group had intact bone trabeculae, abundant collagen fibers, and fewer osteoclasts in femoral tissues; Model group and adenovirus group had sparse bone trabeculae, severe loss of collagen fibers, and more osteoclasts; Bone trabeculae and collagen fibers in overexpression BMP2 adenovirus group were greater than those in model group and adenovirus group, osteoclasts were fewer than those in model group and adenovirus group. The differences of MDA, GSH, SOD, ferritin, and hepcidin in serum in four groups were statistically significant ( P<0.05), among them, MDA and ferritin in model group were higher than those in sham group, GSH, SOD, hepcidin were lower those in sham group, MDA and ferritin levels in overexpression BMP2 adenovirus group were lower those in model group and adenovirus group, GSH, SOD, hepcidin were higher than those in model group and adenovirus group ( P<0.05). BMP2 mRNA expression of femoral tissues in four groups were 1.00±0.10, 0.39±0.12, 0.39±0.08, and 3.46±0.41, respectively, BMP2 protein expression were 0.61±0.06, 0.17±0.06, 0.20±0.05, and 1.03±0.07, respectively, the differences were statistically significant ( P<0.05), among them, model group were lower those in sham group, overexpression BMP2 adenovirus group were higher than those in model group and adenovirus group ( P<0.05). The differences of ALP, OCN, OPN, RANKL, TRAP, CTX-I, hepcidin, FPN1, TfR1, and DMT1 expression of femoral tissues in sham group, model group, adenovirus group, and BMP2 overexpressing adenovirus group were statistically significant ( P<0.05), among them, ALP, OCN, OPN, and hepcidin in model group were lower than those in sham group, RANKL, TRAP, CTX-I, FPN1, TfR1, and DMT1 were higher than those in sham group, ALP, OCN, OPN, and hepcidin in BMP2 overexpressing adenovirus group were higher than those in model group and adenovirus group, RANKL, TRAP, CTX-I, FPN1, TfR1, and DMT1 were lower than those in model group and adenovirus group ( P<0.05). Conclusion:BMP2 intervention iron metabolism related protein expression promotes bone formation in OP rats by increasing hepcidin level.

Objective:To evaluate the effectiveness and safety of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with stage 4/M neuroblastoma (NB).Methods:This study was a prospective, single-arm, multicenter clinical trial conducted by Sun Yat-sen Memorial Hospital, Children′s Hospital of Nanjing Medical University, Children′s Hospital, Zhejiang University School of Medicine, the First Affiliated Hospital of Guangxi Medical University, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. From March, 2019 to August, 2023, 25 children with confirmed with stage 4/M NB and received allo-HSCT were enrolled. The patients received either unrelated cord blood transplantation (UCBT) or peripheral blood stem cell transplantation (PBSCT). Conditioning regimens for UCBT was fludarabine+busulfan+cyclophosphamide+topotecan, and for PBSCT was fludarabine+busulfan+melphalan+thiotepa+antithymocyte globulin, respectively. Until the last follow-up date of September, 2023, the overall survival (OS) rate and event free survival (EFS) rate were analyzed to evaluate efficacy. The engraftment rate and transplant-related complications were statistically assessed to evaluate safety. Survival analysis was performed using the Kaplan-Meier method.Results:Of the 25 patients, there were 15 males and 10 females. The age at transplantation was 5.7 (3.8, 7.3) years. The engraft rate was 100%, with recovery time of neutrophil as 15.7 (12.5, 17.0) d, and the recovery time of platelets as 33.5 (18.0, 48.0) d. Seventeen of the 25 children (68%) developed acute graft versus host disease (aGVHD), occurred at 18.0 (13.0, 22.5) d after transplantation, including 13 of grade Ⅲ-Ⅳ cases. The main sites of aGVHD were skin and intestinal tract. After treatment, 13 cases improved, 4 patients developed chronic graft-versus-host disease (cGVHD). After allo-HSCT, 14 children received maintenance therapy. Twenty of the 25 patients survived, the 2-year cumulative OS rate was (80±9)%, and 2-year EFS rate was (56±11)%. Nine cases (36%) relapsed, the time from allo-HSCT to disease relapse was 10.9 (5.5, 16.0) months. Five cases (20%) died. The hematopoietic stem cell transplantation associated mortality rate was 4% (1/25).The 2-year OS rate of patients who had partial remission prior to allo-HSCT was significant lower than those who had complete remission prior to allo-HSCT ((33±25)% vs. 100%, P=0.037). Conclusion:allo-HSCT is an effective treatment for patients with stage 4/M NB.

Objective:To investigate the efficacy and toxicity of blinatumomab in the first-line and second-line treatment of pediatric B-cell acute lymphoblastic leukemia (B-ALL).Methods:A multi-center retrospective cohort study was conducted to analyze clinical data from 323 pediatric B-ALL patients treated with blinatumomab across 14 hospitals in China from May 2021 to July 2023. Patients were divided into four groups based on the treatment phase and disease status when blinatumomab was used: relapsed/refractory group, post-consolidation minimal residual disease (MRD)-positive group, early MRD-positive group, and MRD-negative group. Blinatumomab for the relapsed/refractory group was considered as second-line treatment, while the other 3 groups as first-line treatment. The MRD negativity rate after treatment, the survival rates and the incidence of severe adverse events were compared across these groups. Patients who received blinatumomab for more than 7 days were included in the efficacy analysis. Survival analysis was performed using the Kaplan-Meier method, and Log-Rank test was used to compare the survival rates among groups.Results:Among the 323 patients, 191 (59.1%) were male, with the age of 6.2 (3.9, 10.5) years. There were 117 patients in the relapsed/refractory group, 62 cases in the post-consolidation MRD-positive group, 43 cases in the early MRD-positive group, and 101 cases in the MRD negative group. In the relapsed/refractory group, the complete remission rate and MRD negativity rate after one course of blinatumomab were 71.4% (35/49) and 81.5% (75/92) for the 49 children without complete remission and the 92 children with flow cytometry-positive MRD, respectively. In the post-consolidation MRD-positive group, the MRD negativity rates after one course of blinatumomab were 100.0% (27/27), 12/16 and 9/19 for patients with MRD positivity detected by flow cytometry, polymerase chain reaction and next-generation sequencing, respectively. In the early MRD-positive group, the MRD negativity rates were 96.7% (29/30) and 9/9 for flow cytometry and next-generation sequencing, respectively. The 2-year overall survival rate and event-free survival rate for the 319 children evaluable for efficacy were (90.6±1.7)% and (87.6±1.9)%, respectively, with the relapsed/refractory group showing significantly lower overall survival rates and event-free survival rate compared to the other groups ( χ2=21.40, 26.21,both P<0.001). Grade 3 or higher adverse events occurred in 128 cases (39.6%), with hematological toxicity observed in 101 cases, while cytokine release syndrome (CRS), infection, and neurotoxicity occurred in 11, 26 and 8 cases, respectively. In addition, there were statistically significant differences in the grade 3 or higher CRS among the four groups ( χ2=8.03, P<0.05). Conclusion:Blinatumomab can clear MRD more effectively and achieve superior survival outcomes when used as first-line treatment for pediatric B-ALL, with less CRS.

Objective:To explore the effect of bone morphogenetic with protein 2 (BMP2) intervention in iron metabolism related protein expression on rats osteoporosis (OP) by regulating hepcidin level.Methods:Thirty-six 8-week-old SPF grade female SD rats with a body weight of 200-220 g were divided into sham group (Removal of adipose tissue around ovaries), model group (ovariectomy was used to prepare postmenopausal OP model rats), adenovirus group (OP model rats injected with adenovirus into the femoral cavity), and overexpression BMP2 adenovirus group (OP model rats injected with BMP2 overexpressing adenovirus into the femoral cavity) using a random number table method, with 9 rats in each group. Rats were euthanized after 2 months of injection, and femoral tissue was taken for micro computed tomography (micro-CT) three-dimensional reconstruction to observe the microstructural characteristics of bone trabeculae, and calculate bone mineral density (BMD), trabecular number (Tb.N), trabecular thickness (Tb.Th), bone volume/total volume (BV/TV), and trabecular separation (Tb.Sp) values. Hematoxylin eosin staining, Masson staining, and tartrate-resistant acid phosphatase (TRAP) staining were used to observe the pathological damage, collagen content, and osteoclast count of femoral tissues in four group rats. Malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), ferritin, hepcidin levels in serum were detected. Real-time quantitative PCR was used to detect BMP2 mRNA expression in femoral tissue. Western blotting were used to detect BMP2, ALP, OCN, OPN, RANKL, TRAP, CTX-I, hepcidin, FPN1, TfR1, DMT1 protein expression in femoral tissue.Results:The differences of BMD, Tb.N, Tb.Th, BV/TV, and Tb.Sp in sham group, model group, adenovirus group, and BMP2 overexpressing adenovirus group were statistically significant ( P<0.05), among them, BMD, Tb.N, Tb.Th, and BV/TV in model group were lower than those in sham group, Tb.Sp was higher than that in sham group, BMD, Tb.N, Tb.Th, and BV/TV in BMP2 overexpressing adenovirus group were higher than those in model group and adenovirus group, Tb.Sp was lower than that in model group and adenovirus group ( P<0.05). Sham group had intact bone trabeculae, abundant collagen fibers, and fewer osteoclasts in femoral tissues; Model group and adenovirus group had sparse bone trabeculae, severe loss of collagen fibers, and more osteoclasts; Bone trabeculae and collagen fibers in overexpression BMP2 adenovirus group were greater than those in model group and adenovirus group, osteoclasts were fewer than those in model group and adenovirus group. The differences of MDA, GSH, SOD, ferritin, and hepcidin in serum in four groups were statistically significant ( P<0.05), among them, MDA and ferritin in model group were higher than those in sham group, GSH, SOD, hepcidin were lower those in sham group, MDA and ferritin levels in overexpression BMP2 adenovirus group were lower those in model group and adenovirus group, GSH, SOD, hepcidin were higher than those in model group and adenovirus group ( P<0.05). BMP2 mRNA expression of femoral tissues in four groups were 1.00±0.10, 0.39±0.12, 0.39±0.08, and 3.46±0.41, respectively, BMP2 protein expression were 0.61±0.06, 0.17±0.06, 0.20±0.05, and 1.03±0.07, respectively, the differences were statistically significant ( P<0.05), among them, model group were lower those in sham group, overexpression BMP2 adenovirus group were higher than those in model group and adenovirus group ( P<0.05). The differences of ALP, OCN, OPN, RANKL, TRAP, CTX-I, hepcidin, FPN1, TfR1, and DMT1 expression of femoral tissues in sham group, model group, adenovirus group, and BMP2 overexpressing adenovirus group were statistically significant ( P<0.05), among them, ALP, OCN, OPN, and hepcidin in model group were lower than those in sham group, RANKL, TRAP, CTX-I, FPN1, TfR1, and DMT1 were higher than those in sham group, ALP, OCN, OPN, and hepcidin in BMP2 overexpressing adenovirus group were higher than those in model group and adenovirus group, RANKL, TRAP, CTX-I, FPN1, TfR1, and DMT1 were lower than those in model group and adenovirus group ( P<0.05). Conclusion:BMP2 intervention iron metabolism related protein expression promotes bone formation in OP rats by increasing hepcidin level.

Objective To investigate the proportion of Th17 cells,Th1-like Th17 cells and FoxP3+Th17 cells in bone marrow of patients with myelodysplastic syndrome(MDS),the expression of interleukin-17A(IL-17A)in bone marrow supernatant and its clinical significance.Methods Forty MDS patients(MDS group)and 18 patients with nutritional anemia(control group)were selected.MDS patients were classified into the low blast(MDS-LB)group(19 cases)and the increased blast(MDS-IB)group(21 cases,including 11 cases of type IB1 and 10 cases of type IB2)based on morphological definition.The MDS patients were scored according to the revised International Prognostic Scoring System(IPSS-R),with 18 cases in the low-risk group(≤4.5)and 22 cases in the high-risk group(>4.5).Flow cytometry was used to detect the proportion of Th17 cells,Th1-like Th17 cells and FoxP3+Th17 cells in bone marrow of the MDS group and the control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of IL-17A in bone marrow supernatant of the above samples.Results The proportion of Th17 cells and the level of IL-17A were higher in patients of the MDS group than those in the control group(P＜0.05).According to the median expression level of IL-17A,the MDS group was divided into the low-expression group(＜13.71 ng/L,20 cases)and the high-expression group(≥13.71 ng/L,20 cases).Compared with the low-expression group,there were higher proportion of patients with blast cells＜5%and low-risk patients(P＜0.05)in the high-expression group.Compared with the IL-17A low-expression group,the IL-17A high-expression group had a higher proportion of patients with blast cells＜5%and relatively low-risk patients(P＜0.05).Compared with the low-risk patients,high-risk patients had a lower proportion of Th17 cells,IL-17A levels and Th1-like Th17 cells,and a higher proportion of FoxP3+Th17 cells(P＜0.05).Compared with the MDS-LB group,the MDS-IB group had a lower proportion of Th17 cells,IL-17A levels and Th1-like Th17 cells,and a higher proportion of FoxP3+Th17 cells(P＜0.05).Conclusion The proportion of Th17 cells and the level of IL-17A are significantly increased in MDS patients.The decreased proportion of Th1-like Th17 cells and the increased proportion of FoxP3+Th17 cells may be related to the increased proportion of blast cells and higher risk stratification in patients.

Objective To analyze the impact of the interval time(IT)between prostate biopsy(PB)and radical prostatectomy(RP)on the perioperative safety and prognostic efficacy of prostate cancer patients.Methods A retrospective analysis was performed on 87 patients who underwent extraperitoneal laparoscopic RP at our hospitals during Jun.2022 to Nov.2024.The patients were divided into the IT ≤2 weeks group(n=42)and the IT ≥4 weeks group(n=45)according to the interval between PB and RP.Baseline data,perioperative indicators,postoperative inflammatory factors,postoperative pathological results,urinary continence,and complication rates were compared between the two groups.Results No statistically significant differences were observed between the two groups in baseline information,operation time,intraoperative blood loss,intraoperative transfusion rate,postoperative hospital stay,catheter removal time,inflammatory factors and complications(P＞0.05).Pathological results showed no significant differences in cancer tissue proportion,positive rate of lymph node,positive rate of surgical margins,and postoperative Gleason scores(P＞0.05).However,the IT ≤2 weeks group exhibited significantly fewer cases of perineural invasion(25 vs.36,59.52％vs.80.00％)and vascular invasion(5 vs.12,11.90％vs.26.67％)compared to the IT≥4 weeks group(P＜0.05).There were no significant differences in postoperative urinary control rate and prostate-specific antigen(PSA)level between the two groups(P＞0.05).Conclusion Radical prostatectomy performed ≤2 weeks after prostate biopsy demonstrates better safety and prognosis.

Objective To evaluate the application of rabbit liver cancer model in teaching interventional medicine for graduate students.Methods A total of 10 first-year master graduate students majoring in Radiological Imaging(Interventional Medicine).who were studying at Zhengzhou University of China,were enrolled in this study.The rabbit liver cancer model was used as the experimental teaching materials.The teaching contents included the establishment of rabbit liver cancer model,the interventional operation of rabbit liver cancer,the method of scientific research and teaching,the evaluation of the teaching effect,and the survey of student satisfaction.Results Under the guidance of teaching tutor,the success rate of VX2 rabbit liver cancer modeling performed by the 10 master graduate students majoring in interventional medicine was 100％,and the mean operational quality assessment score was(11.5±2.0)points.During the operation of interventional surgery,the success rate of femoral artery puncture was also 100％,and the mean score for each interventional operation was(11.8±2.3)points.The students'experimental designs were evaluated by the expert group,the results were as follows:2 cases were rated as excellent,7 cases were rated as good,and one case was rated as moderate.The degree of students'satisfaction with experimental teaching method was high,the specific scores of each item are as follows:the understanding of the rabbit liver cancer model was(4.80±0.40)points,the command of interventional technology was(4.60±0.49)points,and the quality and practicability of teaching materials was(4.90±0.30)points.Conclusion This teaching method of using rabbit liver cancer model experiment can improve the animal experiment ability,interventional operation ability and scientific research innovation ability of graduate students.Animal model teaching method is an innovation of teaching mode for graduate students majoring in interventional medicine.