1.Construction of a system for isolation and purification of NK cells from whole blood donations
Tengyu CAO ; Huayu LIN ; Xuanzhi ZHANG ; Cuimi DUAN ; Yi LIU ; Xiaonan XUE ; Liping SUN ; Yang YU
Chinese Journal of Blood Transfusion 2025;38(2):181-188
[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×108vs (6.49±2.16)×108, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.
2.Correlation Between Traditional Chinese Medicine Syndrome Type of Unstable Angina Pectoris and Iron Metabolism Indices
Jiye CHEN ; Min WU ; Changxin SUN ; Xiaonan ZHANG ; Zeping WANG ; Longtao LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):126-136
ObjectiveTo investigate the correlation between iron metabolism parameters and various syndrome types of unstable angina pectoris (UAP). MethodsA cross-sectional study was conducted from October 2021 to October 2023, encompassing 213 patients diagnosed with UAP at Xiyuan Hospital of Chinese Academy of Chinese Medical Sciences. Additionally, 30 healthy individuals were selected as control cases. Single-factor analysis was used to investigate the differences in clinical data among different Traditional Chinese Medicine (TCM) syndrome types of UAP and their correlation with iron metabolism indices. The study conducted a comparative analysis of the aforementioned clinical data among patients with and without heat-toxic and blood-stasis syndrome. Logistic regression was used to analyze the correlation between TCM syndrome types and related factors. The receiver operator characteristic (ROC) curve was employed to assess the predictive value of iron metabolism indices, along with their sensitivity and specificity. ResultsCompared to those in the control group, serum iron (SI) and serum ferritin (SF) levels were significantly increased in the UAP group (P<0.01), while transferrin (TRF) and total iron binding capacity (TIBC) levels were decreased (P<0.01). However, there was no significant difference in unsaturated iron binding capacity (UIBC). Multivariate binary Logistic regression analysis identified apolipoprotein A1 (ApoA1), homocysteine (HCY), high-sensitivity C-reactive protein (hs-CRP), and SF as independent influencing factors for the UAP patients (P<0.05, P<0.01). Additionally, statistically significant differences were observed in SI, SF, TRF, and TIBC among 213 patients with different TCM types (P<0.01). Patients with heat-toxic and blood-stasis syndrome had higher SI and SF values than those without the syndrome (P<0.01), while their TIBC and TRF values were lower (P<0.01). Multivariate binary logistic regression analysis showed that SI and LDL-C levels were closely associated with the differentiation of heat-toxic and blood-stasis syndrome. ConclusionUAP patients often experience iron metabolism disorders, and the heat-toxic and blood-stasis syndrome are significantly correlated with iron metabolism parameters. The SI and LDL-C levels have high specificity and sensitivity in diagnosing heat-toxic and blood-stasis syndrome.
3.Iodine nutritional status of population before and after system reform of salt industry in Gansu Province
Yugui DOU ; Yanling WANG ; Yongqin CAO ; Wei SUN ; Xiaoyan CHEN ; Jing ZHENG ; Xiaonan ZHU ; Xiulan FEI
Journal of Public Health and Preventive Medicine 2024;35(2):71-74
Objective To understand the impact of system reform of salt industry on iodine nutrition of people in Gansu Province, and to provide a scientific basis for policy adjustment of relevant departments. Methods The investigation period (2014-2021) was divided into two sub-periods: before system reform of salt industry (2014-2016) and after system reform of salt industry (2017-2021). Thirty counties were selected according to the method of “population proportional probability sampling (PPS)” in 2014. According to the iodine deficiency disease monitoring program of Gansu Province, from 2016 to 2021, children aged 8-10 years and pregnant women were taken as research objects to collect urine samples for urine iodine detection. Children in 2014 and 2018 were selected to measure thyroid volume. Results A total of 90 989 children urine iodine samples were investigated, and the median urinary iodine (MUI) of children was 194.70µg/L; 7 663 and 83,326 children's urinary iodine samples were investigated in the two periods, the MUI was 180.73 µg/L and 196.00 µg/L, respectively, and the difference was statistically significant (P<0.05). A total of 44 741 pregnant women's urinary iodine samples were investigated, and the MUI of pregnant women was 176.50 µg/L; 4 480 and 40 261 pregnant women's urinary iodine samples were investigated in the two periods, the MUI was 160.61 µg/L and 178.10 µg/L, respectively, and the difference was statistically significant (P<0.05). The thyroid volume of 1 555 children and 8 509 children was investigated in the two periods, the median thyroid volume was 2.70 mL and 2.55 mL , respectively, and the difference was statistically significant (P<0.05). The rates of goiter in children were 3.15% and 1.26%, respectively, and the difference was statistically significant (P<0.05). Conclusion The iodine nutrition of people in Gansu Province has not fluctuated significantly after the reform of salt industry system and has maintained an appropriate level. It is necessary to pay attention to the potential risk of insufficient iodine nutrition level and thyroid health of key populations such as children and pregnant women and strengthen health education of scientific iodine supplementation.
4.The preliminary application of cinematic rendering reconstruction technology in acute aortic dissection
He ZHANG ; Zhongxiao LIU ; Meng YU ; Miao YU ; Ziyou WANG ; Wenbei XU ; Xiaonan SUN ; Shenman QIU ; Lixiang XIE ; Yanchun ZHANG ; Yankai MENG ; Cunjie SUN ; Kai XU
Journal of Practical Radiology 2024;40(10):1620-1624
Objective To analyze the clinical application value of cinematic rendering(CR)reconstruction technology in acute aortic dissection(AAD),and to compare the imaging quality between CR and volume rendering(VR)reconstruction.Methods Patients with suspected A AD who underwent aortic computed tomography angiography(CTA)were analyzed retrospectively.All images were uploaded to Siemens Syngo.via post-processing workstation for VR and CR three-dimensional reconstruction,respectively.The optimized view angle,staining and transparency were selected and segmented by a radiologist to display the lesion to the full extent.All subjective evaluations of post-processing images were randomly evaluated on Siemens Syngo.via post-processing workstation by two radiologists.The two radiologists reached a consensus after consultation,and the results without consensus were evaluated by another senior radiologist.The 3-point scale was used in the subjective evaluation of post-processing images.The scores of rupture,endometrium,and true and false cavity were recorded.The diagnostic confidence was also recorded.Results A total of 21 ADD patients were enrolled,11 patients(52.3%)were Debakey Ⅲ type.The scores of rupture in CR and VR reconstruction were 2.952 points and 2.619 points,respectively,which had significant difference(P=0.016).For the endometrium of AAD,the score of all 21 patients in the CR reconstruction was 3 points,while only 7 patients(33.3%)in the VR reconstruction had 3 points,which showed significant difference between the both(P<0.001).For the true and false cavity of AAD,only 1 patient(4.8%)in the VR reconstruction was 3 points,while all 21 patients in the CR reconstruction had 3 points(P<0.001).The scores of CR reconstruction on the diagnostic confidence were significantly higher than those of VR reconstruction(P<0.001).Conclusion CR reconstruction can provide photorealistic anatomical post-processing images,and can improve the display and evaluation of AAD.
5.Research progress on the application of fused attention convolutional neural networks in dermatoscopic segmentation
Xiaonan SUN ; Kui LU ; Chen CHEN ; Jiangshan SUN ; Qiyue ZHU
Journal of Shenyang Medical College 2024;26(5):514-523
In automatic skin damage analysis,segmentation is a challenging and critical operation due to factors such as the shape and contrast of hair and skin lesions on the skin.Compared with traditional segmentation methods,deep learning seamlessly integrates feature extraction and task-specific decision-making,achieving segmentation tasks more accurately and efficiently,and effectively reducing the burden and cost of skin cancer screening.This article first introduces the background of dermoscopic segmentation and deep learning models,and introduces the application of deep learning in dermoscopic segmentation.Secondly,this article introduces the algorithm models of convolutional neural networks and attention mechanisms,reviews the application of fused attention convolutional neural networks in dermoscopic segmentation since Jan 2022,and summarizes the improvement strategies,the advantages and disadvantages of the model.The model is further analyzed based on commonly used datasets of dermoscopy and evaluation indicators of image segmentation.Finally,the application of fused attention convolutional neural network in dermoscopic segmentation is summarized and prospected.
6.Capability verification results of urinary iodine laboratories in Gansu Province from 2017 to 2021
Wei SUN ; Yugui DOU ; Yanling WANG ; Jing ZHENG ; Xiaonan ZHU ; Yongqin CAO ; Runping ZHANG
Chinese Journal of Endemiology 2024;43(6):498-501
Objective:To evaluate the testing capability of urinary iodine laboratories in Gansu Province and analyze the existing problems.Methods:Z-score method and uncertainty analysis were used to analyze the external quality control assessment results of urinary iodine laboratories in Gansu Province from 2017 to 2021 (data were collected from Gansu Center for Disease Prevention and Control).Results:From 2017 to 2021, the participation rate in the assessment of urinary iodine laboratories in the province was 100.0% (473/473), the feedback rate was 99.8% (472/473), and the pass rate was 91.9% (434/472). The pass rates for assessment from 2017 to 2021 were 82.7% (62/75), 93.9% (93/99), 94.9% (93/98), 92.0% (92/100), and 94.0% (94/100), respectively. The pass rates for provincial, municipal, and county assessments were 5/5, 98.6% (69/70), and 90.7% (360/397), respectively. The proportions of │Z│≤2, 2 <│Z│ < 3, and│Z│≥3 between laboratories in the province were 84.5% (399/472), 9.3% (44/472), and 6.2% (29/472), respectively. The proportions of│Z│≤2, 2 <│Z│ < 3, and│Z│≥3 within the laboratories were 88.6% (418/472), 9.1% (43/472), and 2.3% (11/472), respectively. There was a significant difference in the composition of │Z│ scores between laboratories annually (χ 2 = 24.60, P = 0.002), the proportion of│Z│≤2 increased from 66.7%(50/75) in 2017 to 90.0% (90/100) in 2021. The│Z│ scores between and within provincial laboratories were both ≤2. The proportion of │Z│≤2 between municipal and county-level laboratories was 91.4% (64/70) and 83.1% (330/397), respectively, the proportion of│Z│≤2 within laboratories was 92.9% (65/70) and 87.7% (348/397), respectively. There was no difference in the composition of│Z│ scores between and within provincial, municipal, and county-level laboratories( P < 0.05). The proportion of two concentration quality control blind sample results in the province that were both within the uncertainty range was 89.2% (421/472). From 2017 to 2021, they were 81.3% (61/75), 91.9% (91/99), 84.7% (83/98), 92.0% (92/100), and 94.0% (94/100), respectively, with statistically significant differences (χ 2 = 9.69, P = 0.021); provinces, cities, and counties were 5/5, 95.7% (67/70), and 87.9% (349/397), respectively, with statistically significant differences (χ 2 = 23.60, P = 0.023). Conclusions:Through continuous external quality control assessments of all established urinary iodine laboratories in Gansu Province, the overall testing capacity of urinary iodine laboratories at all levels has been continuously improved. However, in the future, it is still necessary to strengthen laboratory testing capabilities and improve the level of urine iodine detection.
7.Intratumoral and peritumoral radiomics based on 18F-FDG PET-CT for predicting epidermal growth factor receptor mutation status in lung adenocarcinoma
Jianxiong GAO ; Xinyu GE ; Rong NIU ; Yunmei SHI ; Zhenxing JIANG ; Yan SUN ; Jinbao FENG ; Yuetao WANG ; Xiaonan SHAO
Chinese Journal of Radiology 2024;58(10):1042-1049
Objective:To investigate the value of intratumoral and peritumoral radiomics models based on 18F-FDG PET-CT in predicting epidermal growth factor receptor (EGFR) mutation status in lung adenocarcinoma and interpret peritumoral radiomics features. Methods:This study was a cross-sectional study. Patients with lung adenocarcinoma who underwent 18F-FDG PET-CT at the Third Affiliated Hospital of Soochow University between January 2018 and April 2022 were retrospectively collected and samplied into a training set (309 cases) and a test set (206 cases) in a 6∶4 ratio randomly. Radiomics features were extracted from the intratumoral and peritumoral regions of interest based on PET and CT images, respectively, and the optimal feature sets were selected. Radiomics models were established using the XGBoost algorithm, and radiomics scores (intratumoral CT label, peritumoral CT label, intratumoral PET label, peritumoral PET label) were calculated. Logistic regression analysis was used to construct a clinical model and a combined model (incorporating PET-CT intratumoral and peritumoral radiomics, clinical features, and CT semantic features). The predictive performance of the models was evaluated using receiver operating characteristic curves and the area under the curve (AUC). Unsupervised clustering, Spearman correlation analysis, and visualization methods were used for the interpretability of peritumoral radiomics features. Results:In both the training and test sets, the AUC value of CT peritumoral labels was greater than that of CT intratumoral labels for predicting EGFR mutation status in lung adenocarcinoma (training set: Z=3.84, P<0.001; test set: Z=1.99, P=0.046). In the test set, the AUC value of PET intratumoral labels (0.684) was slightly higher than that of PET peritumoral labels (0.672) for predicting EGFR mutation status, but the difference was not statistically significant ( P>0.05). The combined model had the highest AUC value for predicting EGFR mutation status of lung adenocarcinoma in both the training and test sets and was significantly better than the clinical model (training set: Z=6.52, P<0.001; test set: Z=2.31, P=0.021). Interpretability analysis revealed that CT peritumoral radiomics features were correlated with CT shape features, and there were significant differences in CT peritumoral features between different EGFR mutation statuses. Conclusions:The value of CT peritumoral labels is superior to that of CT intratumoral labels in predicting EGFR mutation status in lung adenocarcinoma. The predictive performance of the model can be improved by combining PET-CT intratumoral and peritumoral radiomics, clinical features, and CT semantic features.
8.Correlation between CT attenuation value of pulmonary artery thrombi and efficacy of interventional thrombolysis in patients with acute pulmonary embolism
Xiaonan SUN ; Zhongxiao LIU ; He ZHANG ; Xin TANG ; Shenman QIU ; Yankai MENG ; Lixiang XIE ; Shaodong LI ; Qingqiao ZHANG ; Kai XU
Chinese Journal of General Practitioners 2024;23(7):728-733
Objective:To analyze the relationship between CT attenuation value of pulmonary artery thrombi and the efficacy of interventional thrombolysis in patients with acute pulmonary embolism (APE).Methods:This was a single center cross-sectional study. The clinical and imaging data of 89 APE patients who underwent interventional thrombolysis in Affiliated Hospital of Xuzhou Medical University from January 2018 to December 2022, were retrospectively analyzed. All patients underwent CT pulmonary angiography (CTPA) before and after thrombolysis, the CT attenuation value of pulmonary artery thrombi and ratio of CT attenuation value of thrombi to left subscapularis muscle CT value were obtained; and the difference of Qanadli embolism index (ΔQ) before and after thrombolysis was calculated. According to the median ΔQ, patients were classified as good efficacy group (ΔQ>50%) and poor efficacy group (ΔQ≤50%). The clinical characteristics and quantitative parameters of CT were compared between the two groups, and the factors associated with efficacy of thrombolysis were analyzed with univariate and multivariate logistic regression. The correlation between CT attenuation value of pulmonary artery thrombi and ΔQ was analyzed by Spearman correlation analysis.Results:The CT attenuation value of thrombi and ratio of attenuation value of thrombi to left subscapularis muscle CT value showed significant difference between the two groups ( P<0.05). Multivariate analysis showed that compared with CT attenuation value of emboli≤53.47 HU, the value>53.47 HU might be associated with the good efficacy of thrombosysis ( OR=9.175, 95% CI: 0.937-89.846, P=0.057). There was a positive correlation between CT value of pulmonary artery thrombi and ΔQ ( r=0.365, P<0.001). Conclusion:The CT attenuation value of thrombi can predict the efficacy of interventional thrombolysis in APE patients, and patients with higher CT attenuation value would have a better treatment response.
9.Establishment and evaluation of a quantitative TaqMan RT-PCR for the detection of SARS-CoV-2 Omicron variants
Yanhong SUN ; Wenlong LI ; Yaoyao CHEN ; Xiaoqing FU ; Xiaonan ZHAO ; Duo LI
Chinese Journal of Microbiology and Immunology 2023;43(5):375-380
Objective:To develop a rapid, simple and cost-effective quantitative TaqMan RT-PCR (RT-qPCR) that could be used as an alternative to sequencing for the detection of Omicron variants and to evaluate its performance.Methods:Primers and TaqMan probes targeting the conserved domains of SARS-CoV-2 ORF1ab and the high-frequency mutation sites in the S gene of Omicron variants were designed. Then a RT-qPCR for the detection of Omicron variants was established. The consistency of the method was verified using samples identified by whole-genome sequencing. The specificity and sensitivity of the method were also evaluated.Results:The established RT-qPCR could distinguish Omicron variants from early epidemic A strains and Alpha and Delta variants of SARS-CoV-2, and the results were consistent with those of whole-genome sequencing with a coincidence rate of 100% (28/28). There was no cross-reactivity with other six respiratory viruses or coxsackievirus group A16. For RNA standards, this method showed good linearity in the range of 10 9-10 3 copies/μl with a correlation coefficient ( R2) greater than 0.99 and detection sensitivity of 10 3 copies/μl. Conclusions:The RT-qPCR designed in this study for Omicron variant detection had good sensitivity and specificity and could be easily performed in laboratories, which would greatly facilitate the monitoring of Omicron variants.
10.A single-center, retrospective analysis of relapse and progression patterns of primary central nervous system lymphoma: can whole brain radiotherapy be replaced?.
Yue QIN ; Rongping LIU ; Xiaonan ZHANG ; Wan ZHANG ; Chen REN ; Dehua WU
Journal of Southern Medical University 2023;43(4):499-506
OBJECTIVE:
To analyze recurrence and progression patterns of primary central nervous system lymphoma (PCNSL) in patients without whole brain radiotherapy (WBRT) and assess the value of WBRT in PCNSL treatment.
METHODS:
This retrospective single-center study included 27 patients with PCNSL, who experienced recurrence/progression after achieving complete remission (CR), partial remission, or stable disease following initial treatments with chemotherapy but without WBRT. The patients were followed up regularly after the treatment for treatment efficacy assessment. By comparing the anatomical location of the lesions on magnetic resonance images (MRI) at the initial diagnosis and at recurrence/progression, we analyzed the patterns of relapse/progression in patients with different treatment responses and different initial status of the lesions.
RESULTS:
MRI data showed that in 16 (59.26%) of the 27 patients, recurrence/progression occurred in out-field area (outside the simulated clinical target volume [CTV]) but within the simulated WBRT target area in 16 (59.26%) patients, and within the CTV (in-field) in 11 (40.74%) patients. None of the patients had extracranial recurrence of the tumor. Of the 11 patients who achieved CR after the initial treatments, 9 (81.82%) had PCNSL recurrences in the out-field area but within WBRT target area; of the 13 patients with a single lesion at the initial treatment, 11 (84.62%) experienced PCNSL recurrence in the out-field area but within WBRT target area.
CONCLUSIONS
Systemic therapy combined with WBRT still remains the standard treatment for PCNSL patients, especially those who achieve CR after treatment or have a single initial lesion. Future prospective studies with larger sample sizes are needed to further explore the role of low-dose WBRT in PCNSL treatment.
Humans
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Lymphoma/radiotherapy*
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Central Nervous System Neoplasms/pathology*
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Retrospective Studies
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Prospective Studies
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Neoplasm Recurrence, Local/drug therapy*
;
Combined Modality Therapy
;
Brain/pathology*
;
Antineoplastic Combined Chemotherapy Protocols/therapeutic use*
;
Methotrexate


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