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.Analysis on Quality Standard of Sennae Folium(Cassia angustifolia) Dispensing Granules Based on Standard Decoctions
Jinxin LI ; Xue DONG ; Shuai DUAN ; Guiyun CAO ; Jinghua ZHANG ; Yongfu LUAN ; Yongqiang LIN ; Xiaodi DONG ; Zhaoqing MENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):192-200
ObjectiveTo establish the quality standards for Sennae Folium(Cassia angustifolia) dispensing granules based on standard decoctions. MethodsHigh performance liquid chromatography(HPLC) specific chromatograms were established for 15 batches of Sennae Folium(C. angustifolia) standard decoctions and 10 of Sennae Folium(C. angustifolia) dispensing granules from different manufacturers, and the similarity evaluation, hierarchical cluster analysis(HCA) and principal component analysis(PCA) were performed. Linear calibration with two reference substances(LCTRS) and quantitative analysis of multi-components by single-marker(QAMS) were established for the common peaks in the specific chromatograms to determine the contents of main components in the decoction pieces, standard decoctions and dispensing granules, and to calculate their transfer rates from decoction pieces to standard decoctions and dispensing granules. ResultsThe similarities of specific chromatograms of 15 batches of Sennae Folium(C. angustifolia) standard decoctions and 10 batches of Sennae Folium(C. angustifolia) dispensing granules were all greater than 0.95, and a total of 8 characteristic peaks were calibrated, and five of them were identified, including kaempferol-3,7-O-diglucoside, apigenin-6,8-di-C-glucoside, quercetin-3-O-gentianoside, sennoside B and sennoside A. HCA and PCA results showed that there were certain differences in the composition of different batches of standard decoctions, but no clustering was observed in the production area. As the standard decoctions, the extract rate of 15 batches of samples was 26.54%-45.38%, the contents of kaempferol-3,7-O-diglucoside, apigenin-6,8-di-C-glucoside, quercetin-3-O-gentianoside, sennoside B and sennoside A were 12.16-19.26, 2.57-4.94, 3.27-5.11, 6.75-11.39, 4.69-7.79 mg·g-1, and their transfer rates from decoction pieces to standard decoctions were 45.41%-79.02%, 29.12%-55.07%, 40.52%-67.90%, 24.72%-49.12%, 27.54%-49.34%, respectively. The extract rates of Sennae Folium(C. angustifolia) dispensing granules(C8-C10) were 38.10%-39.50%, the transfer rates of the above five components from decoction pieces to dispensing granules were 72.85%-73.58%, 53.43%-53.94%, 40.19%-40.74%, 24.62%-25.00%, 28.65%-29.11%, respectively, which were generally consistent with the transfer rates from decoction pieces to standard decoctions. ConclusionCompared with the relative retention time method, LCTRS has higher prediction accuracy and is more suitable for chromatographic columns. The established quality control standard of Sennae Folium(C. angustifolia) dispensing granules based on standard decoction is reasonable and reliable, and all indicators of samples from different manufacturers are within the range specified based on the standard decoction, which can provide reference for the quality control and process research of this dispensing granules.
3.The Impairment Attention Capture by Topological Change in Children With Autism Spectrum Disorder
Hui-Lin XU ; Huan-Jun XI ; Tao DUAN ; Jing LI ; Dan-Dan LI ; Kai WANG ; Chun-Yan ZHU
Progress in Biochemistry and Biophysics 2025;52(1):223-232
ObjectiveAutism spectrum disorder (ASD) is a neurodevelopmental condition characterized by difficulties with communication and social interaction, restricted and repetitive behaviors. Previous studies have indicated that individuals with ASD exhibit early and lifelong attention deficits, which are closely related to the core symptoms of ASD. Basic visual attention processes may provide a critical foundation for their social communication and interaction abilities. Therefore, this study explores the behavior of children with ASD in capturing attention to changes in topological properties. MethodsOur study recruited twenty-seven ASD children diagnosed by professional clinicians according to DSM-5 and twenty-eight typically developing (TD) age-matched controls. In an attention capture task, we recorded the saccadic behaviors of children with ASD and TD in response to topological change (TC) and non-topological change (nTC) stimuli. Saccadic reaction time (SRT), visual search time (VS), and first fixation dwell time (FFDT) were used as indicators of attentional bias. Pearson correlation tests between the clinical assessment scales and attentional bias were conducted. ResultsThis study found that TD children had significantly faster SRT (P<0.05) and VS (P<0.05) for the TC stimuli compared to the nTC stimuli, while the children with ASD did not exhibit significant differences in either measure (P>0.05). Additionally, ASD children demonstrated significantly less attention towards the TC targets (measured by FFDT), in comparison to TD children (P<0.05). Furthermore, ASD children exhibited a significant negative linear correlation between their attentional bias (measured by VS) and their scores on the compulsive subscale (P<0.05). ConclusionThe results suggest that children with ASD have difficulty shifting their attention to objects with topological changes during change detection. This atypical attention may affect the child’s cognitive and behavioral development, thereby impacting their social communication and interaction. In sum, our findings indicate that difficulties in attentional capture by TC may be a key feature of ASD.
4.Quality Evaluation of Chuanxiong Rhizoma Dispensing Granules Based on HPLC Specific Chromatogram and Two Reference Substances for Determination of Multiple Components
Jinxin LI ; Xue DONG ; Shuai DUAN ; Guiyun CAO ; Jinghua ZHANG ; Yongfu LUAN ; Yongqiang LIN ; Xiaodi DONG ; Zhaoqing MENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):246-253
ObjectiveTo establish the specific chromatogram of Chuanxiong Rhizoma dispensing granules(CRdg), and to evaluate its quality by chemometrics and two reference substances for determination of multiple components(TRSDMC). MethodsHigh performance liquid chromatography(HPLC) specific chromatograms were established using 13 batches of CRdg from 7 manufacturers, and preliminary quality evaluation was performed by similarity evaluation and chemometrics analysis. Eight characteristic peaks in the specific chromatogram of CRdg were measured on 22 different types of C18 columns, and the actual retention times were recorded. Taking chlorogenic acid(peak 1) and senkyunolide A(peak 8) as double standard compounds, the retention times of the eight characteristic peaks were predicted by linear calibration using two reference substances(LCTRS), and the method was validated on three other columns of different brands. Taking chlorogenic acid as reference peak, the relative correction factor method(RCFM) was used to quantify cryptochlorogenic acid, caffeic acid, ferulic acid, senkyunolide I and senkyunolide A, and the results were compared with the external standard method(ESM). ResultsThe similarities of specific chromatograms of 13 batches of CRdg were all >0.90, and a total of 8 characteristic peaks were calibrated, and six of them were identified, including chlorogenic acid(peak 1), cryptochlorogenic acid(peak 2), caffeic acid(peak 3), ferulic acid(peak 5), senkyunolide I(peak 6) and senkyunolide A(peak 8). Through chemometric analysis, it was found that ferulic acid, chlorogenic acid, senkyunolide I and cryptochlorogenic acid were the main components causing quality difference in CRdg, and the accuracy of LCTRS in predicting the retention time of 8 characteristic peaks was superior to that of the relative retention time method(RRT). Further comparison of the results obtained from RCFM and ESM showed that there was no statistically significant difference between the two methods. ConclusionA quality evaluation method for CRdg based on HPLC specific chromatogram and TRSDMC is established, its qualitative accuracy is better than that of RRT, the quantitative accuracy is similar to that of ESM, and 4 quality-differentiated components among different manufacturers are found. This method is stable and reliable, and has reference value for the quality evaluation of other dispensing granules.
5.Quality Evaluation of Chuanxiong Rhizoma Dispensing Granules Based on HPLC Specific Chromatogram and Two Reference Substances for Determination of Multiple Components
Jinxin LI ; Xue DONG ; Shuai DUAN ; Guiyun CAO ; Jinghua ZHANG ; Yongfu LUAN ; Yongqiang LIN ; Xiaodi DONG ; Zhaoqing MENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):246-253
ObjectiveTo establish the specific chromatogram of Chuanxiong Rhizoma dispensing granules(CRdg), and to evaluate its quality by chemometrics and two reference substances for determination of multiple components(TRSDMC). MethodsHigh performance liquid chromatography(HPLC) specific chromatograms were established using 13 batches of CRdg from 7 manufacturers, and preliminary quality evaluation was performed by similarity evaluation and chemometrics analysis. Eight characteristic peaks in the specific chromatogram of CRdg were measured on 22 different types of C18 columns, and the actual retention times were recorded. Taking chlorogenic acid(peak 1) and senkyunolide A(peak 8) as double standard compounds, the retention times of the eight characteristic peaks were predicted by linear calibration using two reference substances(LCTRS), and the method was validated on three other columns of different brands. Taking chlorogenic acid as reference peak, the relative correction factor method(RCFM) was used to quantify cryptochlorogenic acid, caffeic acid, ferulic acid, senkyunolide I and senkyunolide A, and the results were compared with the external standard method(ESM). ResultsThe similarities of specific chromatograms of 13 batches of CRdg were all >0.90, and a total of 8 characteristic peaks were calibrated, and six of them were identified, including chlorogenic acid(peak 1), cryptochlorogenic acid(peak 2), caffeic acid(peak 3), ferulic acid(peak 5), senkyunolide I(peak 6) and senkyunolide A(peak 8). Through chemometric analysis, it was found that ferulic acid, chlorogenic acid, senkyunolide I and cryptochlorogenic acid were the main components causing quality difference in CRdg, and the accuracy of LCTRS in predicting the retention time of 8 characteristic peaks was superior to that of the relative retention time method(RRT). Further comparison of the results obtained from RCFM and ESM showed that there was no statistically significant difference between the two methods. ConclusionA quality evaluation method for CRdg based on HPLC specific chromatogram and TRSDMC is established, its qualitative accuracy is better than that of RRT, the quantitative accuracy is similar to that of ESM, and 4 quality-differentiated components among different manufacturers are found. This method is stable and reliable, and has reference value for the quality evaluation of other dispensing granules.
6.Comparative analysis of four risk assessment methods for benzene homologues exposure at key work sites in the automobile manufacturing industry
Lin CHEN ; Danping DUAN ; Zibo CEN ; Peixian CHEN ; Shaofang ZHU ; Hai ZHANG
China Occupational Medicine 2025;52(2):160-165
Objective To explore the applicability of four risk assessment methods in evaluating occupational health risks associated with low-level benzene homologues exposure in key work sites within the automobile manufacturing industry. Methods The work sites (paint mixing and spray painting) with exposure to benzene homologues among six automobile manufacturing enterprises in Guangdong Province were selected as the study subjects using the judgmental sampling method. Qualitative risk assessment, exposure index method, non-carcinogenic risk assessment, and the International Council on Mining and Metals risk rating method were independently applied to evaluate the occupational health risks of benzene homologues at these work sites. Accuracy, consistency, and correlation of the four methods were compared. Results The air levels of benzene, toluene, xylene, and ethylbenzene in work sites of paint mixing and spray painting across all six enterprises met national occupational health standards. The median ratios of occupational exposure limits for benzene and toluene in spray painting site were higher than those in paint mixing site (0.017 vs 0.010, P<0.05). Using this ratio as a reference for evaluating method accuracy, the non-carcinogenic risk assessment method could distinguish paint mixing site from spray painting site in terms of risk level (P<0.05), whereas the other three methods could not (both P>0.05). The result of consistency testing revealed that the Kappa coefficients between the four methods ranged from -0.13-0.26, indicating poor consistency. The result of Spearman correlation analysis showed that the non-carcinogenic risk assessment and exposure index methods had higher correlation with occupational exposure limits ratios (Spearman correlation coefficients were 0.501 and 0.656, respectively, all P<0.05). Conclusion Occupational health risk assessment can serve as a supplementary tool to evaluate the hazard level of key work sites in the automobile manufacturing industry. Non-carcinogenic risk assessment shows higher accuracy.
7.Advances in concurrent chemoradiotherapy plus immunotherapy forunresectable locally advanced non-small cell lung cancer
Lin DUAN ; Jiaxin DAI ; Yulong LIU
Chinese Journal of Radiological Health 2025;34(3):456-462
Locally advanced non-small cell lung cancer (NSCLC) is a highly heterogeneous disease. For patients with driver gene-negative, unresectable NSCLC, traditional treatment primarily involves concurrent chemoradiotherapy. With the advent of the immunotherapy era, integrated treatment strategies combining concurrent chemoradiotherapy with immunotherapy have significantly improved therapeutic outcomes and substantially extended survival periods. This article summarizes the interaction mechanisms and clinical research progress of radiotherapy, chemotherapy, and immune checkpoint inhibitors in the treatment of NSCLC. This review also provides perspectives on the development of therapies in the future and predictive biomarkers, serving as a reference for treatment options for patients with unresectable locally advanced NSCLC.
8.Research advances in stereotactic radiotherapy for brain metastases
Jiaxin DAI ; Lin DUAN ; Rencai FAN ; Yan LING ; Yulong LIU
Chinese Journal of Radiological Health 2025;34(3):463-470
Brain metastases are the most common intracranial tumors, and their incidence is increasing with the improvement of systemic treatments and survival rates. Optimal treatment usually requires a multidisciplinary approach, including radiotherapy, surgical resection, chemotherapy, targeted therapy, and immunotherapy. Stereotactic radiotherapy, compared to whole-brain radiotherapy, offers improved local control rates and reduced risk of neurocognitive impairment, and has become a new standard option for the treatment of brain metastases. Additionally, the widespread use of targeted and immune therapies in brain metastases has significantly improved the survival of some patients. This article reviews and integrates recent literature on the treatment of brain metastases and analyzes the role of stereotactic radiotherapy in comprehensive treatment, aiming to provide a reference for the selection of clinical treatment plans.
9.Clinical Application and Pharmacological Mechanism of Sishenwan in Treatment of Ulcerative Colitis: A Review
Keqiu YAN ; Xiaoyu ZHANG ; Sifeng JIA ; Yuyu DUAN ; Zixing QIAN ; Yifan CAI ; Junyi SHEN ; Wenjie XIAO ; Xinkun BAO ; Guangjun SUN ; Aizhen LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):261-270
Ulcerative colitis (UC), a chronic, non-specific inflammatory bowel disease with typical symptoms such as abdominal pain, diarrhea, and bloody stools, demonstrates a high relapse rate and difficulty in curing. Sishenwan, first recorded in Internal Medicine Abstract (Nei Ke Zhai Yao), are a classic prescription for treating diarrhea caused by deficiency of the spleen and kidney Yang. The core therapeutic principle of Sishenwan is warming and tonifying the spleen and kidney, and astringing the intestine and stopping diarrhea. In recent years, Sishenwan have demonstrated distinct advantages in the clinical treatment of UC. The pathogenesis of UC involves multiple factors, including immune dysregulation and gut microbiota imbalance. Although Western medicine is effective in the short term, its side effects, high relapse rate, and resistance associated with long-term use pose substantial challenges. Sishenwan have shown excellent clinical outcomes in the treatment of UC due to deficiency of the spleen and kidney Yang. Modern clinical studies indicate that Sishenwan, used alone or in combination with Western medicine or other Chinese medicine compound prescriptions, significantly improve the clinical efficacy in treating UC due to deficiency of the spleen and kidney Yang. Sishenwan effectively alleviate core symptoms such as mucus, pus, and blood in stools, and persistent abdominal pain, reduce Mayo scores and the relapse rate, and improve patients' quality of life. Research on the material basis reveals that Sishenwan contain multiple active ingredients such as psoralen, isopsoralen, and evodiamine. Mechanism studies indicate that Sishenwan inhibit the inflammatory cascade reactions by regulating the signal network through multiple targets. Sishenwan regulate cellular immunity and restore intestinal immune homeostasis. At the microecological level, Sishenwan promote the intestinal barrier repair through the "microbiota-metabolism-immunity" axis. The current research still needs to be deepened in aspects such as the mining of specific biomarkers for syndromes and the exploration of the collaborative mechanism of traditional Chinese and Western medicine. In the future, a full-chain system covering syndrome differentiation, targeting, and monitoring needs to be constructed for promoting the paradigm transformation of Sishenwan into precision drugs. This review systematically explains the treatment mechanism of Sishenwan regarding the combination of disease and syndrome and its multi-target regulatory characteristics, providing a theoretical basis and transformation direction for the treatment of UC with integrated traditional Chinese and Western medicine.
10.Clinical Application and Pharmacological Mechanism of Sishenwan in Treatment of Ulcerative Colitis: A Review
Keqiu YAN ; Xiaoyu ZHANG ; Sifeng JIA ; Yuyu DUAN ; Zixing QIAN ; Yifan CAI ; Junyi SHEN ; Wenjie XIAO ; Xinkun BAO ; Guangjun SUN ; Aizhen LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):261-270
Ulcerative colitis (UC), a chronic, non-specific inflammatory bowel disease with typical symptoms such as abdominal pain, diarrhea, and bloody stools, demonstrates a high relapse rate and difficulty in curing. Sishenwan, first recorded in Internal Medicine Abstract (Nei Ke Zhai Yao), are a classic prescription for treating diarrhea caused by deficiency of the spleen and kidney Yang. The core therapeutic principle of Sishenwan is warming and tonifying the spleen and kidney, and astringing the intestine and stopping diarrhea. In recent years, Sishenwan have demonstrated distinct advantages in the clinical treatment of UC. The pathogenesis of UC involves multiple factors, including immune dysregulation and gut microbiota imbalance. Although Western medicine is effective in the short term, its side effects, high relapse rate, and resistance associated with long-term use pose substantial challenges. Sishenwan have shown excellent clinical outcomes in the treatment of UC due to deficiency of the spleen and kidney Yang. Modern clinical studies indicate that Sishenwan, used alone or in combination with Western medicine or other Chinese medicine compound prescriptions, significantly improve the clinical efficacy in treating UC due to deficiency of the spleen and kidney Yang. Sishenwan effectively alleviate core symptoms such as mucus, pus, and blood in stools, and persistent abdominal pain, reduce Mayo scores and the relapse rate, and improve patients' quality of life. Research on the material basis reveals that Sishenwan contain multiple active ingredients such as psoralen, isopsoralen, and evodiamine. Mechanism studies indicate that Sishenwan inhibit the inflammatory cascade reactions by regulating the signal network through multiple targets. Sishenwan regulate cellular immunity and restore intestinal immune homeostasis. At the microecological level, Sishenwan promote the intestinal barrier repair through the "microbiota-metabolism-immunity" axis. The current research still needs to be deepened in aspects such as the mining of specific biomarkers for syndromes and the exploration of the collaborative mechanism of traditional Chinese and Western medicine. In the future, a full-chain system covering syndrome differentiation, targeting, and monitoring needs to be constructed for promoting the paradigm transformation of Sishenwan into precision drugs. This review systematically explains the treatment mechanism of Sishenwan regarding the combination of disease and syndrome and its multi-target regulatory characteristics, providing a theoretical basis and transformation direction for the treatment of UC with integrated traditional Chinese and Western medicine.

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