Objective: To identify an optimal back-flush solution for leukocyte-depleted filters that maximizes peripheral blood mononuclear cell (PBMC) recovery with high viability, long-term storage stability, and sterility of the harvested residues, thereby providing a clinically translatable strategy. Methods: Three sterile bag-packaged solutions—Saline, Solvent, and Hanks' balanced salt solution (HBSS)—were used to back-flush randomly assigned leukocyte-depleted filters. Nucleated cell recovery rate and viability of the harvested residues were compared. The optimal solution identified was applied to an expanded sample set. PBMC viability and yield were evaluated after 1h vs 48h storage of the residues. PBMCs isolated from the residues were cryopreserved in liquid nitrogen for 1 month, followed by post-thaw comparisons of viability and T-cell expansion capacity. Results: The Solvent group achieved the highest and most consistent nucleated cell recovery rate. Post-flush recovery rate from filters after 400 mL whole blood processing was (21.3±1.6)% for the Solvent group, significantly higher than Saline group (19.2±6.3)% and HBSS group (11.2±5.0)%, with residues from all groups maintaining viability >90%. No biologically significant difference in residue viability was observed between 48h vs 1h storage groups (93.3±2.3)% vs (95.7±1.8)%). PBMC recovery rates from residues showed no statistical difference between 48h vs 1h storage groups [(48.2%±9.5%)vs (40.41%±8.35%), P>0.05], with (17.7±2.6)×10 cells. After 1-month cryopreservation and 10-day expansion, PBMCs isolated from 48-hour-stored residues retained (91.2±3.2)% viability and achieved a (61.9±15.9)-fold expansion. Conclusion: The bag-packaged Solvent, as a back-flush solution, enables sterile acquisition of leukocyte-depleted filter residues through closed-system tubing connections. These residues maintained PBMC viability and recovery rates after 48h storage at 2℃-8℃, with post-cryopreservation (1-month liquid nitrogen) viability and expansion capacity remaining stable. This protocol complies with blood bank regulatory criteria, addresses the concerns about the infectious window period in cell therapy raw materials, and provides a clinically translatable strategy for PBMC-based applications.

BACKGROUND:Ca2+expression in astrocytes has been found to be closely related to cognitive function,and the Ca2+signaling pathway regulated by inositol 1,4,5-trisphosphate receptors(IP3R2)and ryanodine receptor(RYR)2 receptors has become a hot spot in the study of cognitive disorder-related diseases. OBJECTIVE:To investigate the expression of Ca2+signals mediated by IP3R2 and RYR2 in hippocampal astrocytes in animal models of delayed encephalopathy after acute carbon monoxide poisoning,and to explore the possible pathogenesis of delayed encephalopathy after acute carbon monoxide poisoning. METHODS:C57BL mice with qualified cognitive function were selected by Morris water maze experiment and randomly divided into control group and experimental group.An animal model of delayed encephalopathy after acute carbon monoxide poisoning was established by static carbon monoxide inhalation in the experimental group,and the same amount of air was inhaled in the control group.Behavioral and neuronal changes,astrocyte specific marker glial fibrillary acidic protein,IP3R2,RYR2 receptor and Ca2+concentration in astrocytes of the two groups were detected using Morris water maze,hematoxylin-eosin staining,western blot,immunofluorescence double labeling and Ca2+fluorescence probe at 21 days after modeling. RESULTS AND CONCLUSION:In the Morris water maze,the escape latency of the experimental group was significantly longer than that of the control group(P<0.05).Hematoxylin-eosin staining results showed that in the experimental group,the number of hippocampal pyramidal cells decreased,the cell structure was disordered,and the nucleus was broken and dissolved.Immunofluorescence results showed that IP3R2 and RYR2 were co-expressed with glial fibrillary acidic protein in the hippocampus,and the expressions of IP3R2,RYR2 and glial fibrillary acidic protein were up-regulated in the hippocampus of the experimental group(P<0.05).Western blot analysis showed that the expressions of IP3R2,RYR2,and glial fibrillary acidic protein in the hippocampus of the experimental group were increased(P<0.05).Ca2+concentration in hippocampal astrocytes increased significantly in the experimental group(P<0.05).To conclude,astrocytes may affect Ca2+signals by mediating IP3R2 and RYR2 receptors,then impair the cognitive function of mice with carbon monoxide poisoning,and eventually lead to delayed encephalopathy after acute carbon monoxide poisoning.

ObjectiveTo observe the clinical efficacy of acupuncture combined with bloodletting and cupping in the treatment of chronic spontaneous urticaria（CSU） and to explore its potential mechanisms of action. MethodsSeventy CSU patients were randomly divided into loratadine group and acupuncture + bloodletting group， with 35 patients in each group. The loratadine group received oral loratadine tablets， 10 mg once daily in the evening. The acupuncture + bloodletting group received acupuncture at Zhongwan （CV 12）， Guanyuan （CV 4）， Tianshu （ST 25）， Zusanli （ST 36）， Sanyinjiao （SP 6）， Xuehai （SP 10）， Quchi （LI 11）， Hegu （LI 4）， Taichong （LR 3）， Baihui （GV 20）， and Shenting （GV 24）， once daily，along with bloodletting and cupping at Dazhui （GV 14） and Geshu （BL 17）， every other day. Both groups were treated for 4 weeks. The 7-day urticaria activity score（UAS7） was assessed before and after the treatment， and levels of serum immunoglobulin E （IgE）， interleukin-4 （IL-4）， interleukin-5 （IL-5）， eosinophil cationic protein （ECP）， plasma tissue factor （TF）， activated factor Ⅶ （FⅦa）， prothrombin fragment 1+2 （F1+2）， D-dimer （D-D） and complement component 5a （C5a） were detected. ResultsA total of 65 patients were included in the final analysis， 32 in the loratadine group and 33 in the acupuncture + bloodletting group. Before treatment， there was no significant difference in UAS7 score， serum IgE， IL-4， IL-5， ECP levels， or plasma TF， FⅦa， F1+2， D-D， C5a levels between groups （P＞ 0.05）. After treatment， both groups showed significant reductions in UAS7 score， serum IgE， IL-4， IL-5， and plasma TF， FⅦa， F1+2， D-D， and C5a levels compared to those before treatment （P＜0.01）. However， after treatment， there was no significant difference in UAS7 score and serum ECP， IgE， IL-4， IL-5 levels between groups （P＞0.05）. The acupuncture + bloodletting group showed lower plasma TF， FⅦa， F1+2， D-D and C5a levels compared to the loratadine group （P＜0.05 or P＜0.01）. ConclusionAcupuncture combined with bloodletting and cupping can effectively improve the skin symptoms of CSU patients and reduce the levels of inflammatory factors. The potential mechanism of action may involve the regulation of the coagulation-complement-mast cell activation axis， thereby inhibiting mast cell degranulation.

ObjectiveTo investigate the feasibility and safety of endoscopic retrograde cholangiopancreatography （ERCP） combined with oral cholangiopancreatography in the treatment of major duodenal papilla gallbladder polyps. MethodsA retrospective analysis was performed for the clinical data of eight patients with choledocholithiasis and gallbladder polyps who underwent ERCP and combined with oral cholangiopancreatography for major duodenal papilla cholecystectomy in Center of Digestive Endoscopy， Jilin People’s Hospital， from May 2022 to June 2024， and related data were collected， including the success rate of surgery， the technical success rate of gallbladder polyp removal， the superselective method of cystic duct， the time of operation， the time of gallbladder polyp removal， and surgical complications. ResultsBoth the success rate of surgery and the technical success rate of gallbladder polyp removal reached 100%， and of all eight patients， three patients used guide wire to enter the gallbladder under direct view， while five patients received oral cholangiopancreatography to directly enter the gallbladder. The time of operation was 51.88±12.34 minutes， and the time of gallbladder polyp removal was 23.13±10.94 minutes. The diameter of gallbladder polyp was 2‍ ‍—‍ ‍8 mm， and pathological examination showed inflammatory polyps in three patients， adenomatous polyps in one patient， and cholesterol polyps in four patients. There were no complications during or after surgery. The patients were followed up for 2‍ ‍—‍ ‍27 months after surgery， and no recurrence of gallbladder polyp was observed. ConclusionOral cholangiopancreatography is technically safe and feasible in endoscopic major duodenal papilla cholecystectomy.

Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.

Objective To investigate the effects of AIDS and tuberculosis(TB)co-infection on CD8+T lymphocytes.Methods Serum CD8+T cells and CD4+T cells were analyzed and compared between human immunodeficiency virus(HIV)alone patients(n=200),TB alone patients(n=200),and HIV/TB co-infection patients(n=200).Results CD8+T cells were significantly higher in HIV alone group compared to HIV/TB co-infection group and TB alone group(x2=128.779,P＜0.001).The level of CD8+T lymphocytes in HIV/TB co-infection group and HIV alone group was higher than normal level(ZHIV/TB=8.343,PHIV/TB＜0.001,ZHIV=7.988,PHIV＜0.001).The CD8+T cells in TB alone group decreased significantly compared with normal levels(ZTB=8.682,PTB＜0.001).The levels of CD4+T cells in the three groups of patients were all lower than normal(ZHIV=11.088,PHIV＜0.001,ZTB=5.562,PTB＜0.001,ZHIV/TB=12.077,PHIV/TB＜0.001).The stratified analysis of CD8+T cell levels by CD4+T cell counts,showed that when CD4+T cell count was≤100 cells/μL,the level of CD8+T cells in HIV alone group was higher than that in TB alone group.HIV alone group had higher CD8+T cells than HIV/TB co-infection group,and the level of CD8+T cells in HIV/TB co-infection group was significantly lower than normal(Z0-100=1.604,P0-100=0.109).When CD4+T cells ≥101 cells/μL,HIV/TB co-infection group had higher CD8+T cells than TB alone group.HIV alone group had higher CD8+T cells than TB alone group.When CD4+T cells＞500 cells/μL,CD8+T cell levels were within the normal range in all three groups of patients.Taking the HIV alone group as a reference,with the decrease of CD4+T cell count,the CD8+T cell count decreased more significantly in TB alone group and HIV/TB co-infection group than in HIV alone group(slope was 0.344 and 0.216,respectively,P＜0.001).Conclusions With the decline of CD4+T cells,both HIV/TB co-infection and TB alone are associated with decrease in CD8+T cells,and the decrease in the TB alone group is more prominent.

Objective To explore the predictive value of the fractional flow reserve derived from coronary computed tomography(FFRCT)in CAD.Methods Clinical data of 136 patients(309 ma-jor coronary vessels)with suspected CAD consecutively attending to Northern Jiangsu People's Hospital from January 2020 to February 2022 were collected and retrospectively analyzed.Coro-nary computed tomography angiography(CCTA)and coronary angiography(CAG)data and FFRCT were collected.The accuracy,positive predictive value,negative predictive value,sensitivity and specificity of CCTA and FFRCT for the diagnosis of stenosis in 309 major coronary arteries were compared;the correlation between FFRCT and CAG was analyzed using the consistency test.Imaging parameters of 156 vessels were analyzed by senior radiologists for smallest luminal diam-eter,stenotic plaque length,stenotic diameter rate,stenotic area rate,stenotic lumen area and plaque properties.The 98 vessels with ≥50％coronary stenosis were assigned into the coronary group,and the 58 vessels with＜50％ischaemic stenosis were into the non-coronary group.Logistic regression was used to analyze the relationship between the vessel parameters of 156 ves-sels and coronary heart disease(CHD);ROC curve was plotted to analyze the value of each pa-rameter and their combination for the diagnosis of CHD.Results The diagnostic efficacy of FFRCT was more accurate than that of CCTA for CAD(P＜0.01),and the Kappa agreement anal-ysis showed that the Kappa value of the diagnostic results of FFRCT and CAG was 0.620,indica-ting high agreement(P＜0.01).Lower smallest luminal diameter,lower FFRCT,longer stenotic plaque length,and higher diameter stenosis rate and area stenosis rate were observed in the coro-nary group than the non-coronary group(P＜0.01).Multivariate logistic regression analysis showed that smallest luminal diameter(OR=0.536,95％CI:0.335-0.858),stenotic plaque length(OR=1.109,95％CI:1.054-1.166),and FFRCT≤0.80(OR=0.204,95％CI:0.078-0.532)were independent influencing factors in predicting CAD(P＜0.01).ROC curve analysis revealed that the AUC value of combined smallest luminal diameter,stenotic plaque length,and FFRCT 0.80 together in diagnosis of CAD was 0.853(0.795-0.918).Conclusion FFRCT has a higher di-agnostic value for CAD than CCTA,and the combined diagnosis of FFRCT≤0.80,smallest luminal diameter,and stenotic plaque length can further improve the accuracy of the diagnosis of CAD.

Objective:To investigate the relationship between lifestyle factors and stroke in the elderly.Methods:Data were obtained from a natural cohort of the Multimodal Intervention to Delay Dementia and Disability in Rural China(MIND-China)study.A total of 5 009 participants aged 60 years and above were enrolled, of whom 954(19.05%)had experienced a stroke.Logistic regression analyses were conducted to assess the association between five lifestyle factors(smoking, drinking, physical exercise, social activity and sleep quality)and the incidence of stroke.Results:In model 1, after adjusting for age, sex and education, only social activity( OR=0.783, 95% CI: 0.668-0.917)and good sleep quality( OR=0.731, 95% CI: 0.621-0.860)emerged as protective factors against stroke(both P<0.05).No statistically significant associations were found between other lifestyle factors and stroke.Analysis of a variety of healthy lifestyle practice combinations showed that having 3( OR=0.639, 95% CI: 0.433-0.944), 4( OR=0.620, 95% CI: 0.409-0.941)and 5( OR=0.397, 95% CI: 0.197-0.799)healthy lifestyle practices were protective factors against stroke(all P<0.05).In model 2, after adjusting for age, sex, education, body mass index, diabetes, hyperlipidemia, hypertension, and the APOE genotype, the results were consistent with those of Model 1. Conclusions:Lifestyle factors are significantly associated with the risk of stroke in the elderly population.The healthier lifestyle practices are adopted, the lower the risk of stroke will be.

Objective To investigate the predictive value of lactate/albumin(L/A)and C-reactive protein(CRP)for 90-day mortality risk in patients with sepsis.Methods Clinical data from 125 patients diagnosed with sepsis in the Comprehensive Intensive Care Unit,Jinhua People's Hospital from February 2021 to February 2024 retrospectively were collected.According to 90-day death or not of patients,they were divided into death group(n=61)and survival group(n=64).Independent risk factors for 90-day mortality in patients with sepsis were analyzed by using multivariate Logistic regression.Receiver operating characteristic(ROC)curve was drawn to analyze the predictive value of L/A,CRP,and L/A combined CRP for 90-day mortality in patients with sepsis.At the same time,subgroup analysis of patients was performed according to the best cut-off value,and Kaplan-Meier survival curves were drawn.Results Multivariate Logistic regression analysis showed that high CRP(OR=1.009,95％CI:1.003-1.015,P=0.006),high L/A(OR=1.449,95％CI:1.050-2.000,P=0.024),and concurrent acute kidney injury(OR=2.890,95％CI:1.198-6.975,P=0.018)were independent risk factors for 90-day mortality in patients with sepsis.Area under the curve(AUC)for predicting 90-day mortality in patients with sepsis by using CRP,L/A,and L/A combined with CRP were 0.645,0.612 and 0.708 respectively.Subgroup analysis showed that 90-day mortality of patients with sepsis in L/A＞1.41 group was significantly higher than that in L/A≤1.41 group(P=0.001),and 90-day mortality of patients with sepsis in CRP＞93.29mg/L group was significantly higher than that in CRP≤93.29mg/L group(P=0.001),which was consistent with the analysis results of Kaplan-Meier survival curve.Conclusion Both L/A and CRP have certain predictive value for 90-day prognosis of patients with sepsis,and their combined predictive effect is better.

Myeloid neoplasms (MNs) belong to a group of hematological malignancies characterized by the abnormal biological functions of hematopoietic stem progenitor cells. The abnormal immune and hematopoietic microenvironment of patients with MN interact with malignant clonal hematopoietic stem cells, promoting the occurrence and development of their diseases. MN large granular lymphocyte proliferation (MN-LGLP) is a special and rare clinical phenomenon in this type of disease. Currently, research on this disease in domestic and international cohorts is limited. This study analyzes the clinical and laboratory characteristics of this type of patient and explores the impact of LGLP on the clinical characteristics and survival of patients with MN. Patients with MN-LGLP are prone to neutropenia and splenomegaly. The presence of LGLP is not a risk factor affecting the survival of patients with MN-LGLP. STAG, ASXL1, and TET2 are the most common accompanying gene mutations in MN-LGLP, and patients with MN-LGLP and STAG2 mutations have poor prognoses.