1.Mechanisms of Dihuang Yinzi in Treating Advanced Parkinson's Disease Based on Gut Microbiota-SCFAs-inflammation Axis
Renzhi MA ; Yasi LIN ; Tingyue JIANG ; Hongmei ZHU ; Jiayuan LI ; Yu WANG ; Ge ZHANG ; Wenxin FAN ; Jinli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):11-21
ObjectiveTo observe the effects of Dihuang Yinzi (DY) on motor dysfunction in rats with advanced Parkinson's disease (PD) and to investigate the mechanisms by which DY improves advanced PD symptoms through the "gut microbiota-short-chain fatty acids (SCFAs)-inflammation-neuroprotection pathway". MethodsAn advanced PD rat model was induced by rotenone. Rats were divided into a normal group, model group, positive drug group (levodopa, 50 mg·kg-1), and DY low-, medium-, and high-dose groups (5.2, 10.4, 20.8 g·kg-1). After 7 days of administration, motor function was evaluated using the open-field, pole-climbing, and inclined plate tests. Hematoxylin-eosin (HE) staining was used to observe pathological changes in the substantia nigra and colon, and immunohistochemistry was performed to detect α-Synuclein (α-Syn) and tyrosine hydroxylase (TH) expression in the substantia nigra. Enzyme-linked immunosorbent assay (ELISA) was used to measure levels of dopamine (DA), 5-hydroxytryptamine (5-HT), 3,4-dihydroxyphenylacetic acid (DOPAC), Levodopa, homovanillic acid (HVA), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β). Western blot analysis was used to detect the expression of zonula occludens-1 (ZO-1) and occludin. Gut microbiota diversity was analyzed by 16S rRNA sequencing, and gas chromatography (GC) was used to determine the content of SCFAs in colonic contents. ResultsCompared with the normal group, the model group showed significantly decreased movement speed and distance in the open-field test, prolonged pole-climbing time, and reduced retention angle on the inclined plate (P<0.01), accompanied by increased α-Syn expression (P<0.01) and decreased TH expression (P<0.01) in the brain. Compared with the model group, all DY dose groups improved motor dysfunction in advanced PD rats to varying degrees (P<0.05, P<0.01) and alleviated pathological damage in the brain and colon. High-dose DY significantly reduced α-Syn aggregation in the substantia nigra (P<0.01) and increased TH expression (P<0.01). ELISA and Western blot results showed that, compared with the normal group, the model group exhibited decreased levels of DA, 5-HT, DOPAC, Levodopa, and HVA in the striatum (P<0.01), increased levels of TNF-α, IL-6, and IL-1β in the colon and striatum (P<0.01), and significantly reduced expression of ZO-1 (P<0.05) and occludin in the colon (P<0.01). Compared with the model group, all DY dose groups increased the levels of DA, 5-HT, DOPAC, Levodopa, and HVA in the striatum to varying degrees (P<0.05, P<0.01). In the high-dose DY group, the levels of TNF-α, IL-6, and IL-1β in the colon and striatum were reduced (P<0.01), while the expression of ZO-1 (P<0.05) and occludin in the intestine was increased. The 16S rRNA sequencing results indicated that the relative abundances of Actinobacteriota, Enterobacteriaceae, and Erysipelotrichaceae were increased in the model group, whereas the relative abundances of Bacteroidota, class Clostridia, Lachnospiraceae, and Akkermansia muciniphila were decreased. These changes were effectively reversed after high-dose DY intervention. GC analysis showed that the content of SCFAs in the colonic contents of rats in the model group was decreased (P<0.05, P<0.01), while after high-dose DY intervention, the levels of acetate, propionate, isobutyrate, and butyrate were significantly increased (P<0.05, P<0.01). ConclusionDY may exert therapeutic effects in advanced PD by regulating the gut microbiota-SCFAs-inflammation pathway.
2.Herbal Textual Research on Bambusae Succus in Famous Classical Formulas
Yu SHI ; Feng ZHOU ; Yihan WANG ; Yanmeng LIU ; Ming YANG ; Zhiping CHEN ; Jiangshan ZHANG ; Conglong XU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):231-239
This article systematically reviews and examines the historical evolution of Bambusae Succus as a medicinal material, covering aspects such as nomenclature, origin, geographical distribution, harvesting and processing methods, quality assessment, therapeutic effects and indications, by consulting ancient herbal texts, medical compendia, and modern literature. The aim is to provide a reference for the development and utilization of famous classical formulas containing this herb. Research indicated that Bambusae Succus was first documented in the Shennong Bencaojing during the Han dynasty, with Zhuli being the standard name used throughout history, alongside aliases like Zhuzhi, Zhuyou and Huoquan. Historically, the primary source of Bambusae Succus has been Phyllostachys nigra var. henonis(Danzhu), although other species such as Pleioblastus amarus and Bambusa emeiensis have also been used medicinally. Ancient records predominantly noted its origin in Yizhou(present-day Chengdu and surrounding areas in Sichuan) and the Wuling region(between present-day Hunan, Guangdong, Guangxi and Jiangxi provinces), while contemporary sources are mainly from regions south of the Yangtze River and southwestern China. Traditionally, Bambusae Succus was harvested from bamboo that had grown for exactly one year, today, it can be collected year-round without strict age requirements. Ancient preparation methods included direct fire roasting or dry distillation, whereas modern industrial production employs dry distillation, reflux extraction, and percolation. In terms of quality evaluation, ancient texts considered a sweet taste to be superior, while today, clarity and transparency are prioritized. Historically, Bambusae Succus was characterized as sweet and cold nature, targeting the lung and stomach meridians, with uses evolving from clearing heat and resolving phlegm to nourishing Yin, moistening dryness, and relaxing tendons and unblocking meridians. Modern descriptions classify it as sweet, bitter, and cold in nature, affecting the heart, liver, and lung meridians, with functions including clearing heat, resolving phlegm, and facilitating orifices. It is indicated for conditions such as stroke with phlegm confusion, lung heat with phlegm congestion, convulsions, epilepsy, excessive phlegm in febrile diseases, high fever with thirst, irritability during pregnancy, and tetanus, with more clearly defined applications. Based on the results of the research, it is recommended that when developing and utilizing famous classical formulas containing Bambusae Succus, the one-year-old Phyllostachys nigra var. Henonis, which has been highly praised throughout history, should be selected as the source material. Industrial production should adopt the dry distillation method. Furthermore, in-depth research should be conducted on the modern technological characterization of the traditional quality control indicator of sweet taste, and reasonable modern quality control standards should be established.
3.Optimized monovision Q-value-customized FS-LASIK for myopia, astigmatism, and presbyopia
Linjuan YANG ; Qiang SHI ; Zhao LIU ; Yu ZHANG ; Shengjian MI
International Eye Science 2026;26(5):772-779
AIM: To evaluate safety, efficacy, visual quality, and patient satisfaction after optimized monovision Q-value-customized femtosecond laser-assisted in situ keratomileusis(FS-LASIK)for myopia, astigmatism, and presbyopia. METHODS: This prospective study enrolled patients suffering myopia, astigmatism, and presbyopia. At 1 y post operation, uncorrected distance, intermediate and near visual acuity(UDVA, UIVA, UNVA), refraction, corneal higher order aberrations(HOAs), Q value, objective visual quality, and near stereoacuity were evaluated. The impact of surgery on patients' living quality and their satisfaction were evaluated based on National Eye Institute Refractive Error Quality of Life Instrument(NEI RQL)scores at 1-year follow-up.RESULTS:The study enrolled 36 patients(72 eyes)including 12 males(33%)and 24 females(67%)with an average age of 46.03±3.60(range 40-53)y. One year postoperatively, 97% of patients achieved binocular UDVA of 1.0 or better, 89% achieved UIVA of 0.63 or better, and 97% achieved UNVA of 1.0 or better. The spherical equivalent refraction(SE)in dominant eye within ±1.00 D was 100%. SE in non-dominant eyes was uncorrected compared with the preoperative target correction, with a mean difference of -0.27±0.34 D before and after surgery(P<0.001). while in the non-dominant eyes, the anterior corneal Q-value within the 5 mm zone increased negatively compared with the preoperative value(-0.33±0.24 vs -0.21±0.09, P<0.001). In the non-dominant eye, spherical aberration and longitudinal spherical aberration were lower than preoperative values(both P<0.001). Besides, there were no significant changes in objective scatter index(OSI), modulation transfer function cutoff frequency(MTF cut off)and Strehl ratio(SR)in both dominant and non-dominant eyes(all P>0.05), and the near stereoacuity was better after surgery(P=0.007). Additionally, the patient satisfaction rate with the surgery was 94%, the satisfaction rates for UDVA and UNVA were both 98%. The 11% of patients required to wear low-diopter glasses when driving at night.CONCLUSION:Optimized monovision Q-value-customised FS-LASIK is a safe and effective procedure for correcting myopia, astigmatism, and presbyopia. It provides favorable subjective and objective visual quality and high patient satisfaction. The under correction in non-dominant eyes is observed, indicating the further optimization of nomogram is needed.
4.Exploring Biological Characteristics of Rat Model of Atrial Fibrillation with Phlegm-heat and Blood Stasis Pattern Based on Metabolomics
Ailin HOU ; Yuxuan LIU ; Wenxi YU ; Xing JI ; Chan WU ; Dazhuo SHI ; Ying ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):245-255
ObjectiveTo establish an animal model of atrial fibrillation(AF) that accurately reflects the phlegm-heat and blood stasis(TRYZ) pathogenesis in traditional Chinese medicine. MethodsForty SPF-grade SD rats were randomly assigned using a random number table to the following groups:the control group, the TRYZ+AF group,the AF group and the TRYZ group, with ten rats in each group. The TRYZ+AF and TRYZ groups underwent a high-fat diet combined with intraperitoneal lipopolysaccharide(LPS) injection to simulate the pathological alterations of TRYZ syndrome. Groups TRYZ+AF and AF were induced with acetylcholine-calcium chloride(Ach-CaCl2) via caudal vein injection to induce AF. The control group received no intervention and was maintained under normal conditions. The modeling period lasted 3 weeks. Electrocardiography was used to assess AF episodes and duration, echocardiography evaluated left atrial dimensions and cardiac function, fully automated biochemical analyzer measured the levels of total cholesterol(TC), triglycerides(TG), high-density lipoprotein cholesterol(HDL-C) and low-density lipoprotein cholesterol(LDL-C), hemoreometer analyzed the whole blood viscosity, plasma viscosity, and whole blood reduced viscosity, a coagulation analyzer assessed prothrombin time(PT), activated partial thromboplastin time(APTT), thrombin time(TT), and fibrinogen(FIB), enzyme-linked immunosorbent assay(ELISA) was used to determine the levels of C-reactive protein(CRP), interleukin(IL)-1β, IL-6, IL-17, tumour necrosis factor(TNF)-α, matrix metalloproteinase-9(MMP-9), galectin-3(Gal-3), Collagen Ⅰ, and α-smooth muscle actin(α-SMA). Hematoxylin-eosin(HE) staining and Masson's trichrome staining were used to analyze pathological changes in atrial myocardium, Western blot was employed to detect MMP-9, Collagen Ⅰ and α-SMA protein expression in myocardial tissue, real-time quantitative polymerase chain reaction(Real-time PCR) evaluated fibrous factor gene expression levels. Changes in the TRYZ syndrome were assessed via body weight, tongue color[red(R), green(G), and blue(B)], and rectal temperature. Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was employed to detect differential metabolites between the control group and the TRYZ+AF group. ResultsFollowing three weeks of sustained modeling, compared with the control group, rats in the TRYZ+AF and the TRYZ groups exhibited reduced body weight, dry faeces, elevated rectal temperature, dark red tongue, decreased RGB values on the tongue surface, and markedly elevated TC and LDL-C levels(P<0.05, P<0.01). The TRYZ+AF, TRYZ, and AF groups exhibited significantly decreased TT, APTT and PT, along with markedly elevated whole blood viscosity and FIB(P<0.05, P<0.01). Rats in the TRYZ+AF and AF groups exhibited AF rhythm, markedly decreased heart rate, prolonged RR intervals, enlarged left atrium, and significantly reduced ejection fraction and shortening fraction(P<0.05, P<0.01). Serum levels of CRP, IL-1β, IL-6, IL-17, TNF-α, MMP-9, Gal-3, Collagen Ⅰ, and α-SMA were elevated in rats from the TRYZ+AF, TRYZ, and AF groups compared to the control group, with the most pronounced increase observed in the TRYZ+AF group(P<0.05, P<0.01). Histopathology revealed that the collagen fiber deposition in the atrial of rats in the TRYZ+AF, TRYZ and AF groups was higher than that in the control group(P<0.05, P<0.01). Western blot and Real-time PCR results further demonstrated that the protein and mRNA expression levels of MMP-9, Collagen Ⅰ and α-SMA in the myocardial tissue of the TRYZ+AF group were higher than those in the other three groups(P<0.05, P<0.01). Metabolomic analysis revealed 173 differentially expressed metabolites in the TRYZ+AF group and the control group, primarily enriched in pathways such as glycerophospholipid metabolism and glycolysis/gluconeogenesis. ConclusionThis study successfully establishes a rat model of AF integrated with the TRYZ syndrome, demonstrating the pathological process where the interactions of phlegm, heat and stasis jointly trigger tremor, this provides a reliable experimental tool for in-depth research into the biological basis of this disease syndrome.
5.Research progress on the mechanisms of traditional Chinese medicine in treating functional constipation based on the gut microbiota-bile acid axis
Xiangrui KONG ; Qimeng ZHANG ; Yue ZOU ; Yong LIANG ; Yu SHI ; Yang ZHANG ; Hongxi ZHANG
China Pharmacy 2026;37(2):244-249
Functional constipation (FC) is a common functional disorder of the intestines, mainly characterized by reduced bowel movement frequency, difficulty in defecation, a sensation of incomplete evacuation, and hard stools, which severely affect patients’ quality of life. Research indicates that the pathogenesis of FC is closely related to gut microbiota dysbiosis and abnormal bile acid secretion. Bile acids, as endogenous natural laxatives, promote bowel movements by enhancing colonic secretion and regulating intestinal motility; meanwhile, gut microbiota influence colonic transit function by regulating the enteric nervous system, immune system, and their metabolic products. Based on an overview of the relationship between gut microbiota and bile acid metabolism, this article systematically reviews the current research status on the mechanisms of traditional Chinese medicine (TCM) in treating FC by regulating the balance of the gut microbiota-bile acid axis. It is found that single Chinese medicinal herbs (such as Atractylodes macrocephala), isolated compounds (such as Platycodon grandiflorum polysaccharides), herbal formulas (such as Shanger huang pill), acupuncture, and moxibustion can up-regulate the abundance of beneficial bacteria, reshape the microbial structure, correct bile acid metabolism, and activate the Takeda G-protein receptor 5/farnesoid X receptor pathway to treat FC.
6.Research progress on traditional Chinese medicine regulation of MAPK signaling pathway in intervening slow transit constipation
Xiangrui KONG ; Qimeng ZHANG ; Yue ZOU ; Yong LIANG ; Yu SHI ; Yang ZHANG ; Ke MENG ; Hongxi ZHANG
China Pharmacy 2026;37(11):1508-1514
low transit constipation (STC) is a common functional intestinal disorder caused by impaired colonic transit function, characterized by reduced bowel movement frequency, hard stools, and difficulty in defecation. The mitogen-activated protein kinase (MAPK) signaling pathway, which mainly includes extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 subtypes, plays a critical regulatory role in the occurrence and development of STC. This paper systematically reviews the multiple pathogenic mechanisms of the MAPK signaling pathway in STC and the research progress of traditional Chinese medicine (TCM) intervention.At the mechanistic level, the MAPK signaling pathway promotes the progression of STC through the following links:(1) Activation of p38 upregulates the expression of aquaporin 3 (AQP3)/AQP4 in the colon, leading to excessive reabsorption of water in the intestinal lumen; (2) It forms a positive feedback loop with nuclear factor-κB (NF-κB) to maintain low-grade intestinal inflammation, releases inflammatory factors such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), and inhibits smooth muscle contraction; (3) Overactivation of p38 downregulates the expression of occludin and mucin 2 while upregulates the expression of claudin-2, thereby disrupting the mucosal barrier; (4) The JNK/p38 signaling pathway activates the caspase cascade to induce apoptosis of intestinal epithelial cells, neurons, and interstitial cells of Cajal; (5) Abnormal ERK signaling and excessive activation of p38/JNK inhibit intestinal smooth muscle contraction and reduce 5-hydroxytryptamine secretion, ultimately resulting in impaired colonic transit function.At the intervention level, TCM compound formulas and single herbs have been proven to improve STC by regulating the MAPK signaling pathway. Their effects are syndrome type-dependent:yin-nourishing formulas (Zengye Chengqi Tang, Tongbian Tang) mainly regulate the ERK/AQP axis; yang-warming formulas (Jichuan Jian) target both ERK/JNK and anti-apoptosis; heat-clearing formulas (Sanren Tang) focus on p38/NF-κB anti-inflammation. A single drug can simultaneously cover multiple aspects including water metabolism, inflammation, barrier function, apoptosis, and intestinal motility.Current relevant studies still have limitations such as mechanisms mostly remaining at the correlational level and a lack of disease-syndrome integrated research models. Future studies should combine specific inhibitors or gene knockout to identify core targets, establish disease-syndrome integrated STC models, and use network pharmacology and molecular docking techniques to deeply analyze the fine mechanism of “component-target-phenotype”, so as to provide high-quality evidence for the precise regulation of the MAPK signaling pathway by TCM in the intervention of STC.
7.Expert consensus on the application of artificial intelligence in lung cancer screening, diagnosis, and treatment (2026 edition)
Wenzhao ZHONG ; Haibo WANG ; Yi HU ; Hao ZHANG ; Jigang DAI ; Junqiang FAN ; Guibin QIAO ; Fan YANG ; Jian HU ; Fengwei TAN ; Xuening YANG ; Qiang PU ; Zihao CHEN ; Hongxia TIAN ; Lunxu LIU ; Hecheng LI ; Xiaolong YAN ; Zongyang YU ; Zhenbin QIU ; Yihua SUN ; Jing HU ; Yuhang SHI ; Zhifei GUO ; Peng ZHANG ; Kezhong CHEN ; Shugeng GAO ; Yilong WU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(06):848-856
With the continuous deepening of the concept of precision diagnosis and treatment for lung cancer, how to achieve higher efficiency and accuracy in the screening, diagnosis, and treatment pathways in clinical practice has become an important issue that urgently needs to be overcome. The current clinical difficulty lies in the fact that despite continuous advancements in imaging and molecular diagnostic technologies, there are still limitations in manual efficiency and subjective experience when it comes to massive data analysis and multi-scale feature extraction. Artificial intelligence (AI), especially algorithm systems based on deep learning, is an innovative technology capable of deeply empowering medical big data. This method utilizes algorithms such as convolutional neural networks, combined with radiomics, pathomics, and multi-modal data fusion analysis, demonstrating immense potential in early precise detection and benign-malignant differentiation of pulmonary nodules, digital pathological subtype recognition and non-invasive prediction of driver genes, precise 3D surgical planning and automatic delineation of radiotherapy target volumes, as well as dynamic risk warning during follow-up. This innovative technology provides a brand-new solution for realizing intelligent and individualized lung cancer diagnosis and treatment models. This consensus, based on the latest evidence from evidence-based medicine and combined with the development trends in the AI field and real-world clinical needs, was ultimately formed by gathering the consensus opinions of multidisciplinary experts in radiology, pathology, thoracic surgery, and other fields. The main content covers the application specifications of AI in the three core scenarios of lung cancer screening, diagnosis, and treatment, the technical standards for data collection and algorithm validation, as well as the ethical and regulatory challenges faced at the current stage. It aims to clarify the applicable boundaries of AI as a clinical auxiliary decision support tool, providing scientific guidance and standardized exploration directions for peers currently engaged in or planning to carry out AI-assisted clinical diagnosis, treatment, and translation of lung cancer.
8.A Personalized Brain-computer Interface Paradigm and Decoding Method for The Objective Evaluation of Auditory Frequency Difference Limen
Sheng-Ye LI ; Xiao-Lin XIAO ; Shi-Hang YU ; Bei-Bei ZHANG ; Xing-Wei AN ; Min-Peng XU ; Dong MING
Progress in Biochemistry and Biophysics 2026;53(7):1927-1941
ObjectiveThe frequency difference limen (FDL) serves as a fundamental metric utilized for effectively quantifying the precise perceptual capabilities of the central auditory system. However, traditional measurement methods rely heavily on the active behavioral responses of subjects and are consequently highly susceptible to the negative influence of confounding subjective factors. Furthermore, existing research paradigms frequently employ uniform stimulus configurations that overlook critical individual perceptual differences. Based on brain-computer interface (BCI) technology, this comprehensive study aims to establish an objective and quantitative evaluation method for auditory frequency discrimination by systematically analyzing and decoding the specific neural responses elicited at the exact threshold state. MethodsWe designed a personalized rapid serial auditory presentation (RSAP) paradigm customized based on each individual’s precise FDL. A cohort of eleven healthy participants was recruited to evaluate the paradigm using pure-tone sequences at a baseline frequency of 4 000 Hz. This experimental paradigm simulates a realistic auditory perception environment through the continuous presentation of acoustic stimuli, thereby allowing for an in-depth investigation into the specific neural representations evoked by weak frequency deviations at the threshold state. Given that auditory stimulus-evoked response features exhibit complex and differentiated spatiotemporal distribution patterns across multiple frequency domains, this study further deeply integrates the cross-scale feature interaction module with the dynamic spatiotemporal attention allocation strategy, innovatively proposing the Multi-Scale Spatial-Temporal Dual Attention Network (MS-STAMNet). Specifically, the network constructs parallel processing branches with multiple receptive fields and introduces a dynamic adaptive weighting strategy to precisely localize core neural activity signals, further deeply integrating multi-scale information through cross-branch feature information interaction to achieve robust single-trial decoding of weak auditory evoked responses. ResultsThe comprehensive electrophysiological data analysis demonstrated that subtle auditory frequency deviation stimuli presented at the threshold level successfully elicited pronounced N2 and P3 event-related potential features, reflecting pre-attentive mismatch detection and subsequent cognitive evaluation, which were prominently distributed over the frontal, central, and temporal regions of the scalp. In the complex time-frequency domain, the extracted neural response characteristics exhibited distinct, statistically significant event-related synchronization within both the low-frequency δ and θ frequency bands, which was simultaneously accompanied by a widespread, prominent event-related desynchronization within the higher α band. A comparative analysis of model performance demonstrated that MS-STAMNet achieved an average unweighted average recall (UAR) of (69.67±6.12)% and area under the curve (AUC) of 0.761 8±0.07, significantly outperforming the established baseline models such as EEGNet and PLNet. Furthermore, a distinct dissociation phenomenon was verified between neural decoding and behavioral performance through regression analysis (R2=0.016, P=0.709), indicating that this model can effectively capture the implicit features of subtle frequency deviations, even when they fail to trigger explicit conscious responses. Additionally, attention weight visualization analysis further reveals the highly accurate focus of the network on key features concentrated over the bilateral temporal and fronto-parietal regions. ConclusionThis study systematically and comprehensively uncovers the multi-dimensional spatiotemporal evolutionary patterns of complex neural responses processing subtle acoustic variations under long-sequence threshold auditory stimulation. Concurrently, it verifies the efficacy and robustness of the proposed MS-STAMNet architecture in accurately deciphering weak, single-trial electroencephalogram signals amidst complex background noise. Ultimately, these neurophysiological and algorithmic findings lay a solid theoretical and methodological foundation for the objective and quantitative evaluation of individual auditory cognitive capabilities in clinical applications, transcending the fundamental limitations of traditional behavioral paradigms and providing robust technical support for future auditory research and related clinical assessments.
9.Process optimization for recycling immune cells from discarded leukocyte filter and research on NK cell culture
Xuebing ZHOU ; Ruyi LI ; Hong YUAN ; Xuesong ZHANG ; Yu SHI ; Xu YANG ; Bairui DOU ; Lixin JIAO ; Yu HAN
Chinese Journal of Blood Transfusion 2026;39(7):846-852
Objective: To optimize the technology and process for recovering immune cells from discarded leukocyte filter through reverse flushing, and to investigate the effects of different reverse flushing solutions and physical conditions (temperature, soaking, and ultrasound) on the cell recovery rate. Additionally, to evaluate the feasibility and quality of isolating and culturing NK cells from the recovered cells. Methods: Normal saline, PBS, and PBS-EDTA were used as reverse flushing solutions to backwash the filter, and the white blood cell recovery rates were compared. The reverse flushing efficiency was optimized by adjusting the temperature of the reverse flushing solution, filter soaking treatment and ultrasonic treatment. The Ficoll separation method was employed to isolate peripheral blood mononuclear cells (PBMCs) from the elution solution and conduct in vitro expansion and culture. Lymphocyte subsets were identified and quantified using flow cytometry. Results: The physiological saline served as the backflush fluid was the most stable. The reverse temperature had a significant impact on the recovery rate of white blood cell subgroups. The recovery rates of lymphocytes and monocytes at 37℃ were significantly higher than those at 4℃ [(35.56±5.55)% vs (22.88±0.31)%, (32.42±4.82)% vs (8.00±1.6)%, t=5.581 and 5.758, respectively, both P<0.001)], and at 25℃ [(35.56±5.55)% vs (22.42±6.13)%, t=5.904, P<0.001; (32.4±4.82)% vs (21.11±1.23)%, t=2.629, P=0.030)]. The recovery rates of white blood cell subgroups after 37℃ immersion were higher than those after direct reverse flow at 37℃, but the difference was not statistically significant (P>0.05). The recovery rate of cells after ultrasonic immersion at 25℃ was further increased compared to that after immersion at 25℃, but the difference was not statistically significant (P>0.05). The purity of the NK cells obtained through sorting reached (94.2±4.5)%, and the viability was (94.8±0.9)%. After 15 days of culture, the cell count was expanded approximately 450 times, yielding more than 10
/mL highly active NK cells. Conclusion: By optimizing the backflushing process, immune cells can be efficiently recovered from discarded filters. The 37℃ backflow liquid has the best backflow effect; soaking and ultrasonic treatment can be used as auxiliary methods to improve recovery rates. The NK cells from this source have high purity, good activity, and strong expansion ability, and can be used as a reliable cell source for immunotherapy research, and in line with the circular economy concept of resource reuse.
10.Contamination status and dietary exposure risk assessment of arsenic, cadmium, lead and mercury in commercially available aquatic products in Shanghai
Rong SHI ; Dandan MAO ; Limei CHEN ; Yan ZHANG ; Huimin ZHANG ; Yu GAO ; Ying TIAN
Shanghai Journal of Preventive Medicine 2026;38(8):598-603
ObjectiveTo investigate the contamination status of arsenic (As), cadmium (Cd), lead (Pb), and mercury (Hg) in commercially available aquatic products in Shanghai and their distribution characteristics among different categories of aquatic products, and to assess the dietary exposure levels and non‑carcinogenic health risks of As, Cd, Pb, and Hg through aquatic product consumption among different age groups, so as to provide a scientific basis for food safety surveillance of aquatic products and dietary risk management for residents in Shanghai. MethodsFrom October to November 2021, a total of 90 aquatic product samples, including marine fish, freshwater fish, freshwater shrimp, marine crab, freshwater crab, and mollusks, were randomly collected from 16 districts of Shanghai. Sample pretreatment was performed according to GB 5009.268‒2016 (National Food Safety Standard—Determination of Multi‑elements in Foods). The concentrations of As, Cd, and Pb in aquatic products were determined by inductively coupled plasma mass spectrometry (ICP‑MS), and Hg was determined by direct mercury analyzer. Inorganic arsenic (iAs) was estimated as 3.5% of total As according to the method recommended by the European Food Safety Authority (EFSA). The contamination levels of As, Cd, Pb, and Hg in aquatic products were evaluated according to GB 2762‒2017 (National Food Safety Standard—Maximum Levels of Contaminants in Foods). A combined approach of point assessment and probabilistic assessment was applied to estimate the estimated daily intake (EDI) and non‑carcinogenic risk indicators—hazard quotient (HQ) and hazard index (HI)—for different age groups. ResultsThe detection rates of As and Hg were both 100.00% in all samples, while those of Cd and Pb were 74.44% and 25.56%, respectively. The arithmetic means (wet weight) of the four elements and estimated iAs were: As (1.432 9±3.736 5) mg·kg-1, iAs (0.050 2±0.130 8) mg·kg-1, Cd (0.082 5±0.197 8) mg·kg-1, Pb (0.006 1±0.005 5) mg·kg-1, and Hg (0.021 9±0.016 7) mg·kg-1. According to the relevant maximum residue limits, 2 out of 7 mollusk samples (28.57%) and 3 out of 20 marine fish samples (15.00%) exceeded the limit for iAs, and 2 out of 8 marine crab samples (25.00%) exceeded the limit for Cd. The estimated daily intake (EDI, μg·kg-1·d-1, by body weight) of iAs, Cd, Pb, and Hg via aquatic product consumption varied among different age groups, with the highest levels observed in children aged 3‒4 years, decreasing gradually with age to the lowest level at 12 ‒ <15 years, with a slight rebound thereafter. The point assessment results showed that the HQs of iAs, Cd, Pb, and Hg ranged from 0 to 0.25, and the cumulative HI ranged from 0.27 to 0.48, all below 1, indicating a relatively low overall risk. The probabilistic assessment results revealed that under the high‑exposure scenario (P97.5), the HQs of iAs (1.101‒1.281) and Cd (1.175‒1.366) exceeded 1 in children aged 3‒9 years, suggesting potential health risks, while the HQs of Pb and Hg were all below 1, indicating relatively lower risks. ConclusionHg and As are ubiquitously detected in commercially available aquatic products in Shanghai, while the detection rates of Cd and Pb are relatively low. The overall non-carcinogenic risk is at an acceptable level, but children aged 3‒9 years may have potential health risks from iAs and Cd under high-percentile exposure levels. It is recommended to strengthen the monitoring of toxic elements in key aquatic products such as marine fish, mollusks, and marine crabs, and to implement targeted dietary risk management measures for sensitive populations, including children.

Result Analysis
Print
Save
E-mail