Objective To establish and evaluate an integrated disease-syndrome rat model of chronic cerebral ischemia with Qi deficiency and blood stasis syndrome.Methods Thirty male Wistar rats were allocated randomly into three groups(n=10 per group):sham operation(sham),2-vessel occlusion(2-VO)group,and sleep deprivation combined with 2-VO(SD+2-VO)group.We comprehensively assessed Qi deficiency and blood stasis syndrome manifestations in the rats using a dual evaluation approach,combining exhaustive swimming tests with quantitative tongue chroma analysis.Cognitive function was evaluated using the Barnes maze,and cerebral blood flow was compared using laser speckle contrast imaging.The histopathology of the hippocampal cytoarchitecture and white matter were examined using hematoxylin-eosin(HE)and Luxol fast blue(LFB)staining,respectively,and ultrastructural alterations of neurons in the hippocampal CA1 region were observed by transmission electron microscopy(TEM).Protein expression levels of NeuN,vascular endothelial growth factor A(VEGFA)and CD31 were detected by Western Blot and immunofluorescence.Results Cerebral blood flow was significantly reduced in rats in the 2-VO group compared with the sham group,but they failed to recapitulate the key clinical hallmarks of Qi deficiency and blood stasis syndrome.In contrast,rats in the SD+2-VO group exhibited significantly reduced locomotor activity,exacerbated cerebral hypoperfusion,shortened swimming duration,and darkened tongue color compared with 2-VO rats.Rats in the SD+2-VO group demonstrated significantly impaired learning and memory abilities in the Barnes maze test.Consistent with these observations,HE staining,TEM,and LFB staining revealed substantial neuronal and white matter damage in the SD+2-VO group.NeuN expression was decreased and VEGFA and CD31 expression levels were increased in the 2-VO and SD+2-VO groups,as shown by Western Blot.Taken together,these findings indicated that the SD+2-VO model effectively recapitulated the clinical features of chronic cerebral ischemia with Qi deficiency and blood stasis pattern.Conclusions The combination of sleep deprivation and bilateral carotid artery occlusion successfully established a rat model of chronic cerebral ischemia with Qi deficiency and blood stasis syndrome.Compared with the 2-VO model,SD+2-VO model demonstrates more pronounced syndrome manifestations and better clinical relevance,thus providing a valuable animal model for traditional Chinese medicine research on chronic cerebral ischemia.

Objective:To design a novel model for experiments on in vitro irradiation with radioactive seeds using a treatment planning system (TPS) and 3D printing technology and to preliminarily validate the design scientific rigor of the model via experiments on isodose brachytherapy (BT) and external beam radiotherapy (EBRT) on glioma cells in mice. Methods:The TPS was employed to design the model′s shape and calculate the number and positions of radioactive seeds, and 3D printing technology was utilized to fabricate the experimental model. The GL261 cell line was selected for in vitro irradiation experiments, with the mice divided into the control, EBRT, and BT groups. Mice in the EBRT and BT groups were treated with EBRT and BT, respectively, at doses of 2, 4, and 6 Gy. Then, changes in their cell viability, proliferation, and the level of intracellular reactive oxygen species (ROS) were assessed. Results:The model for in vitro irradiation with radioactive seeds was successfully designed and fabricated. The single photon emission computed tomography (SPECT) verified a uniform radioactive distribution within the model, with no significant cold spots. The BT and EBRT groups displayed decreased cell viability with an increase in the radiation dose. Compared to the EBRT group, the BT group exhibited significantly reduced cell viability (51.33% vs. 22.00%, t = 10.94, P < 0.05) and clone counts (172.67 ± 13.11 vs. 53.67 ± 10.22, t = 8.73, P < 0.05), but a significantly increased level of ROS (102.52 ± 6.87 vs. 144.81 ± 6.01, t = -5.26, P < 0.05) at a dose of 6 Gy. Conclusions:An effective model of in vitro irradiation with radioactive seeds is designed based on TPS and 3D printing technology. This provides an experimental model tool and target for research on the BT and EBRT mechanisms.

Objective:To employ single-photon emission computed tomography (SPECT)/CT for isodose assignment in 125I brachytherapy, assess the correlation between photon counts and dose values, and develop a clinical γ-ray visualization model for 125I brachytherapy. Methods:125I radioactive seeds were filled into a self-made 3D printed stereotactic template to build a stereotactic model. The model was scanned by SPECT/CT for photon counts at 0.5, 1.0, 1.5, 2.0 cm from the outermost peripheral seeds, and the corresponding dose values were measured using the Treatment Planning System (TPS). The fitting curve for the photon counts and the dose values was plotted using SPSS 27.0 software. Results:The photon counts of γ rays at distances of 0.5, 1.0, 1.5, and 2.0 cm from the peripheral particles were 7 603.57±1 806.35, 4 018.26±1 315.72, 2 074.04±791.53, and 1 080.34±424.79, respectively, showing a significant difference ( F=743.72, P<0.01). The dose values (in Gy) in the TPS at distances of 0.5, 1.0, 1.5, and 2.0 cm from the peripheral particles were 208.05±37.57, 125.43±17.74, 86.76±17.67, and 61.55±14.39, respectively, which were significantly different ( F=930.46, P<0.01). The photon counts were linearly correlated with the dose values ( y=0.02 x+ 46.45, R2=81.2%, P<0.01). Conclusions:SPECT/CT-based γ-ray photon count detection can be used to assign doses for 125I brachytherapy, enabling the visualization of γ rays in 125I brachytherapy. This approach has a distinct advantage over TPS, laying the foundation for the establishment of an alternative system to TPS.

To introduce the general thinking, guidelines, work objectives and elaboration process of the general technical requirements adopted by volume Ⅳ of the Chinese Pharmacopoeia 2025 Edition, and to summarize and figure out the main characteristics on dosage forms, physico-chemical testing, microbial and biological testing, reference standards and guidelines The newly revised general chapters of pharmacopoeia give full play to the normative and guiding role of the Chinese Pharmacopoeia standard, track the frontier dynamics of international drug regulatory science and the elaboration of monographs, expand the application of state-of-the-art technologies, and steadily promote the harmonization and unification with the ICH guidelines； further enhance the overall capacity of TCM quality control, actively implement the 3 R principles on animal experiments, and practice the concept of environmental-friendly； replace and/or reduce the use of toxic and hazardous reagents, strengthen the requirements of drug safety control This paper aims to provide a full-view perspective for the comprehensive, correct understanding and accurate implementation of general technical requirements included in the Chinese Pharmacopoeia 2025 Edition.

To introduce the general thinking,guidelines,work objectives and elaboration process of the general technical requirements adopted by volume Ⅳ of the Chinese Pharmacopoeia 2025 Edition,and to summarize and figure out the main characteristics on dosage forms,physico-chemical testing,microbial and biological testing,ref-erence standards and guidelines.The newly revised general chapters of pharmacopoeia give full play to the norma-tive and guiding role of the Chinese Pharmacopoeia standard,track the frontier dynamics of international drug regu-latory science and the elaboration of monographs,expand the application of state-of-the-art technologies,and stead-ily promote the harmonization and unification with the ICH guidelines;further enhance the overall capacity of TCM quality control,actively implement the 3 R principles on animal experiments,and practice the concept of environ-mental-friendly;replace and/orreduce the use of toxic and hazardousreagents,strengthen the requirementsofdrug safety control.This paper aims to provide a full-view perspective for the comprehensive,correct understanding and accurate implementation of general technical requirements included in the Chinese Pharmacopoeia 2025 Edition.

Objective:To design a novel model for experiments on in vitro irradiation with radioactive seeds using a treatment planning system (TPS) and 3D printing technology and to preliminarily validate the design scientific rigor of the model via experiments on isodose brachytherapy (BT) and external beam radiotherapy (EBRT) on glioma cells in mice. Methods:The TPS was employed to design the model′s shape and calculate the number and positions of radioactive seeds, and 3D printing technology was utilized to fabricate the experimental model. The GL261 cell line was selected for in vitro irradiation experiments, with the mice divided into the control, EBRT, and BT groups. Mice in the EBRT and BT groups were treated with EBRT and BT, respectively, at doses of 2, 4, and 6 Gy. Then, changes in their cell viability, proliferation, and the level of intracellular reactive oxygen species (ROS) were assessed. Results:The model for in vitro irradiation with radioactive seeds was successfully designed and fabricated. The single photon emission computed tomography (SPECT) verified a uniform radioactive distribution within the model, with no significant cold spots. The BT and EBRT groups displayed decreased cell viability with an increase in the radiation dose. Compared to the EBRT group, the BT group exhibited significantly reduced cell viability (51.33% vs. 22.00%, t = 10.94, P < 0.05) and clone counts (172.67 ± 13.11 vs. 53.67 ± 10.22, t = 8.73, P < 0.05), but a significantly increased level of ROS (102.52 ± 6.87 vs. 144.81 ± 6.01, t = -5.26, P < 0.05) at a dose of 6 Gy. Conclusions:An effective model of in vitro irradiation with radioactive seeds is designed based on TPS and 3D printing technology. This provides an experimental model tool and target for research on the BT and EBRT mechanisms.

Objective:To employ single-photon emission computed tomography (SPECT)/CT for isodose assignment in 125I brachytherapy, assess the correlation between photon counts and dose values, and develop a clinical γ-ray visualization model for 125I brachytherapy. Methods:125I radioactive seeds were filled into a self-made 3D printed stereotactic template to build a stereotactic model. The model was scanned by SPECT/CT for photon counts at 0.5, 1.0, 1.5, 2.0 cm from the outermost peripheral seeds, and the corresponding dose values were measured using the Treatment Planning System (TPS). The fitting curve for the photon counts and the dose values was plotted using SPSS 27.0 software. Results:The photon counts of γ rays at distances of 0.5, 1.0, 1.5, and 2.0 cm from the peripheral particles were 7 603.57±1 806.35, 4 018.26±1 315.72, 2 074.04±791.53, and 1 080.34±424.79, respectively, showing a significant difference ( F=743.72, P<0.01). The dose values (in Gy) in the TPS at distances of 0.5, 1.0, 1.5, and 2.0 cm from the peripheral particles were 208.05±37.57, 125.43±17.74, 86.76±17.67, and 61.55±14.39, respectively, which were significantly different ( F=930.46, P<0.01). The photon counts were linearly correlated with the dose values ( y=0.02 x+ 46.45, R2=81.2%, P<0.01). Conclusions:SPECT/CT-based γ-ray photon count detection can be used to assign doses for 125I brachytherapy, enabling the visualization of γ rays in 125I brachytherapy. This approach has a distinct advantage over TPS, laying the foundation for the establishment of an alternative system to TPS.

Objective To establish and evaluate an integrated disease-syndrome rat model of chronic cerebral ischemia with Qi deficiency and blood stasis syndrome.Methods Thirty male Wistar rats were allocated randomly into three groups(n=10 per group):sham operation(sham),2-vessel occlusion(2-VO)group,and sleep deprivation combined with 2-VO(SD+2-VO)group.We comprehensively assessed Qi deficiency and blood stasis syndrome manifestations in the rats using a dual evaluation approach,combining exhaustive swimming tests with quantitative tongue chroma analysis.Cognitive function was evaluated using the Barnes maze,and cerebral blood flow was compared using laser speckle contrast imaging.The histopathology of the hippocampal cytoarchitecture and white matter were examined using hematoxylin-eosin(HE)and Luxol fast blue(LFB)staining,respectively,and ultrastructural alterations of neurons in the hippocampal CA1 region were observed by transmission electron microscopy(TEM).Protein expression levels of NeuN,vascular endothelial growth factor A(VEGFA)and CD31 were detected by Western Blot and immunofluorescence.Results Cerebral blood flow was significantly reduced in rats in the 2-VO group compared with the sham group,but they failed to recapitulate the key clinical hallmarks of Qi deficiency and blood stasis syndrome.In contrast,rats in the SD+2-VO group exhibited significantly reduced locomotor activity,exacerbated cerebral hypoperfusion,shortened swimming duration,and darkened tongue color compared with 2-VO rats.Rats in the SD+2-VO group demonstrated significantly impaired learning and memory abilities in the Barnes maze test.Consistent with these observations,HE staining,TEM,and LFB staining revealed substantial neuronal and white matter damage in the SD+2-VO group.NeuN expression was decreased and VEGFA and CD31 expression levels were increased in the 2-VO and SD+2-VO groups,as shown by Western Blot.Taken together,these findings indicated that the SD+2-VO model effectively recapitulated the clinical features of chronic cerebral ischemia with Qi deficiency and blood stasis pattern.Conclusions The combination of sleep deprivation and bilateral carotid artery occlusion successfully established a rat model of chronic cerebral ischemia with Qi deficiency and blood stasis syndrome.Compared with the 2-VO model,SD+2-VO model demonstrates more pronounced syndrome manifestations and better clinical relevance,thus providing a valuable animal model for traditional Chinese medicine research on chronic cerebral ischemia.

Abstract: Objective To conduct genotyping and molecular tracing analysis on Plasmodium vivax samples from a cluster of P. vivax malaria outbreak in order to provide a reference for case geographical origin determination. Methods Blood samples from 4 patients in a vivax malaria cluster in Longhui County, Hunan Province from June to July 2018 were collected for species identification by qPCR, and 9 microsatellite molecular markers were used to genotype the parasite strains from four samples. The population genetic STRUCTURE analysis was performed based on the VivaxGEN-MS microsatellite genotype database of P. vivax in the Asia Pacific Malaria Elimination Network, to determine the genetic subgroups and geographical origin of the strains. Results By qPCR, all 4 cases were identified as Plasmodium vivax infection, and 9 microsatellite loci of the 4 cases were successfully typed, and the four samples had different genetic haplotypes, among which case 1, case 3, and case 4 were infected by a single clonal strain, and case 2 was infected by a polyclonal strain. When all P. vivax samples were divided into 2 subpopulations (K=2) by STURCTURE analysis, 4 Hunan samples were classified into tropical genetic subpopulations (comprising strains from Ethiopia, Iran, Bhutan, Malaysia, Indonesia, and southern China). When the samples were divided into 4 subgroups by STURCTURE analysis (K=4), the 4 Hunan samples were classified as South Asian/Southeast Asian genetic subgroups (originating from Bhutan, Malaysia, Indonesia, and southern China). Conclusions The results of molecular tracing do not support that the 4 P. vivax strains in this outbreak originated from the population of central China. The technology of molecular tracing of P. vivax can provide objective evidence for determining the source of infection in malaria cases during the stage of malaria elimination and post-elimination.

Objective:To explore the influencing factors of hemodialysis (HD) initiation in non-diabetic kidney disease (NDKD) patients with predialysis arteriovenous fistula (AVF) creation.Methods:This was a single-center prospective cohort study. The NDKD patients undergoing predialysis AVF creation were enrolled at the First Affiliated Hospital of Xi'an Jiaotong University from 2015 to 2018. According to the estimated glomerular filtration rate (eGFR, the Chronic Kidney Disease Epidemiology Collaboration equation) and age, patients were divided into different subgroups, eGFR: group 1 [eGFR<10 ml·min -1·(1.73 m 2) -1], group 2 [ eGFR between 10 to 15 ml·min -1·(1.73 m 2) -1], and group 3 [eGFR > 15 ml·min -1·(1.73 m 2) -1]; age: age ≥65 years group and age <65 years group. The primary outcome was defined as the initiation of HD within 1 year after AVF surgery. The second outcome was the use of AVF access at the time of HD initiation. Cox proportional hazard regression was performed to identify which demographic and clinical factors were associated with the initiation of HD after AVF surgery. Logistic regression analysis was performed to investigate factors associated with AVF use at the initiation of HD. Results:A total of 220 patients were enrolled, with age of (48.1±16.2) years, of which 143(65.0%) were males. Overall, the clinical parameters of eGFR, cystatin C, serum albumin, 24h-Urine protein, serum phosphorus were as follows respectively, 7.7 (6.6,9.2) ml·min -1·(1.73 m 2) -1, (3.93±1.12) mg/L, (36.0±4.0) g/L, (2.22±1.36) g, (1.71±0.53) mmol/L. The proportion of patients initiating HD within 6 months ( Fisher=6.832, P=0.020) and the level of hemoglobin ( F=3.112, P=0.047) were higher in group 3 compared to the other two eGFR groups. While the median time interval between AVF creation and HD initiation ( H=6.295, P=0.043) was shorter in group 1. In age <65 years group, the level of serum albumin ( t=2.076, P=0.039), triglyceride ( t=1.995, P=0.048) were higher compared with age ≥65 years group; interestingly, the proportion of patients initiated HD within 3 months ( χ2=4.033, P=0.045) and 6 months ( χ2=5.012, P=0.025) were lower in age <65 years group. The median time interval between AVF creation and HD initiation among these patients was 84 (49,174) days. The patients initiating HD within 3 months, 6 months, and 1 year after AVF creation were 112 (50.9%), 152 (69.1%), and 202 (91.8%), respectively. Multivariate Cox regression analysis indicated that higher cystatin C level ( HR=1.283, 95% CI 1.121-1.469, P<0.001) was associated with earlier HD initiation within 1 year of AVF surgery in NDKD patients. AVF usage was accomplished in 64.3% of patients who initiated HD within 90 days, the ratio was 100.0% in those initiated HD between 91 to 180 days, and 88.0% in those ≥181 days after AVF surgery. No factor was independently associated with AVF use at HD initiation identified by multivariate logistic regression analyses in patients with NDKD. Conclusion:Serum cystatin C level is associated with HD initiation within 1 year of the predialysis AVF creation in NDKD patients.