1.Multicenter machine learning-based construction of a model for predicting potential organ donors and validation with decision curve analysis
Xu WANG ; Wenxiu LI ; Fenghua WANG ; Shuli WU ; Dong JIA ; Xin GE ; Zhihua SHAN ; Tongzuo LI
Organ Transplantation 2026;17(1):106-115
Objective To evaluate the predictive value of different machine learning models constructed in a multicenter environment for potential organ donors and verify their clinical application feasibility. Methods The study included 2 000 inpatients admitted to five domestic tertiary hospitals from January 2020 to December 2023, who met the criteria for potential organ donation assessment. They were randomly divided into a training set and an internal validation set (7∶3). Another 300 similar patients admitted to the First Affiliated Hospital of Harbin Medical University from January 2024 to April 2025 were included as an external validation set. The area under the curve (AUC), sensitivity, specificity, accuracy and F1-score of three models were compared, and the consistency of the potential organ donor determination process was tested. Multivariate logistic regression analysis was used to identify predictive factors of potential organ donors. Decision curve analysis (DCA) was employed to verify the resource efficiency of each model, and the threshold interval and intervention balance point were assessed. Results Apart from age, there were no significant differences in other basic characteristics among the centers (all P>0.05). The consistency of the potential organ donor determination process among researchers in each center was good [all 95% confidence interval (CI) lower limits >0]. In the internal validation set, the XGBoost model had the best predictive performance (AUC=0.92, 95% CI 0.89-0.94) and the best calibration (P=0.441, Brier score 0.099). In the external validation set, the XGBoost model also had the best predictive performance (AUC=0.91, 95% CI 0.88-0.94), outperforming logistic regression and random forest models. Multivariate logistic regression showed that mechanical ventilation had the greatest impact (odds ratio=2.06, 95% CI 1.54-2.76, P<0.001). DCA indicated that the XGBoost model had the highest net benefit in the threshold interval of 0.2-0.6. The “treat all” strategy only had a slight advantage at extremely low thresholds. The recommended threshold interval, which balances intervention costs and clinical benefits, considers ≥50% positive predictive value (PPV) and ≤50 referrals per 100 high-risk patients. Conclusions The XGBoost model established in a multicenter environment is accurate and well-calibrated in predicting potential organ donors. Combined with DCA, it may effectively guide the timing of clinical interventions and resource allocation, providing new ideas for the assessment and management of organ donation after brain death.
2.Effect and Mechanism of Liangyi Paste on Hepatic Lipid Deposition in Naturally Aged Mice with High-fat Diet via Cuproptosis/Oxidative Stress Pathway
Meiling ZHANG ; Yuanguang DONG ; Xiaofei SUN ; Jiaxin WANG ; Yu LIU ; Jingxuan ZHU ; Qun WANG ; Nan SONG ; Guoyuan SUI ; Lianqun JIA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):91-99
ObjectiveTaking the cuproptosis/oxidative stress pathway as the entry point, this study investigated the effect and mechanism of Liangyi Paste on hepatic lipid deposition in naturally aged mice fed with a high-fat diet. MethodsAfter adaptive feeding, 80 ten-week-old male C57BL/6 mice were used. Thirty of them were randomly divided into three groups (10 mice per group): The 12-month-old control group (12MCON), the 15-month-old control group (15MCON), and the 15-month-old group with a high-fat diet (15MHFD). The 12MCON and 15MCON groups were continuously fed a standard diet, while the 15MHFD group started receiving a high-fat diet at 12 months of age. Tissue samples were collected at the corresponding time points for each group. The remaining 50 mice were randomly divided into five groups (10 mice per group): the 20-month-old control group (20MCON), the model group, and the low-, medium-, and high-dose Liangyi Paste groups (2.91 , 5.82 , 11.64 g·kg-1·d-1, respectively). The 20MCON group was continuously fed a standard diet, while the other groups started receiving a high-fat diet at 15 months of age. At 18 months of age, the Liangyi Paste groups were administered the corresponding doses of Liangyi Paste by gavage, while the 20MCON and model groups were given an equal volume of saline by gavage. After 8 weeks of continuous gavage (when the mice reached 20 months of age), tissue samples were collected. Hepatic TG levels were measured using assay kits; liver histology and lipid deposition were observed via hematoxylin-eosin (HE) and oil red O staining; reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay (ELISA); Cu2+, superoxide dismutase (SOD), and malondialdehyde (MDA) levels were measured by colorimetry; mRNA and protein expression of genes related to cuproptosis and oxidative stress pathways were analyzed by Real-time polymerase chain reaction(Real-time PCR) and Wes automated protein expression system. ResultsCompared with 12MCON, the 15MCON group showed significantly increased hepatic TG, Cu2+, ROS, and MDA levels (P<0.01), decreased SOD (P<0.01), hepatocyte swelling, and disordered arrangement. The mRNA and protein levels of ferredoxin 1 (FDX1), dihydrolipoamide S-acetyltransferase (DLAT), heat shock protein 70 (HSP70), dihydrolipoamide dehydrogenase (DLD), pyruvate dehydrogenase E1 subunit-β (PDHB), nuclear factor erythroid 2-related factor 2 (Nrf2), and peroxisome proliferator-activated receptor γ (PPARγ) were significantly elevated (P<0.05, P<0.01). Compared with 15MCON group, the 15MHFD and 20MCON groups exhibited further increases in TG, Cu2+, ROS, and MDA (P<0.01), reduced SOD (P<0.01), and aggravated hepatocyte swelling and disorder. There were increased lipid droplets with mild vacuolization in the 15MHFD group, and no significant lipid deposition was observed in the 20MCON group. FDX1, DLAT, HSP70, DLD, PDHB, Nrf2, and PPARγ mRNA and protein levels were significantly increased (P<0.05, P<0.01). Compared with 20MCON group, the model group demonstrated markedly elevated TG, Cu2+, ROS, and MDA (P<0.01), reduced SOD (P<0.01), severe hepatic steatosis, and upregulated expression of FDX1, DLAT, HSP70, DLD, PDHB, Nrf2, and PPARγ mRNA and proteins (P<0.05, P<0.01). All abnormalities were significantly reversed after Liangyi Paste treatment. ConclusionLiangyi paste can ameliorate hepatic lipid deposition in naturally aged mice with a high-fat diet by modulating the cuproptosis/oxidative stress pathway.
3.Compact Fundus Imaging System Using Shack-Hartmann Wavefront Sensing for High-speed Auto-focus
Zhe-Kai LIN ; Long CHEN ; Geng-Yong ZHENG ; Jin-Tian HUANG ; Jia-Xin DONG ; Shang-Pan YANG ; Wen-Zheng DING ; Ding-An HAN ; Xue-Hua WANG ; Ya-Guang ZENG
Progress in Biochemistry and Biophysics 2026;53(4):1076-1086
ObjectiveThe widespread adoption of portable fundus cameras for primary care and community screening is hindered by limitations in current autofocus(AF) technologies. Image-based methods relying on sharpness evaluation require iterative searches, resulting in slow convergence, while projection-based techniques are susceptible to optical artifacts and calibration errors. To address these challenges, this study introduces a novel AF system based on direct wavefront sensing, designed to deliver simultaneous high speed, high precision, and operational robustness within the compact form factor essential for portable ophthalmic devices. MethodsOur approach fundamentally reimagines the AF process by directly measuring the ocular wavefront aberration. We developed a custom portable fundus camera integrating a miniaturized Shack-Hartmann wavefront sensor (SHWS) into the optical path. An 850 nm laser diode projects a point source onto the retina via oblique illumination to minimize corneal reflections. Light scattered from this spot carries the eye’s refractive error through the imaging optics and is directed to the SHWS, positioned at a plane optically conjugate to the primary color CMOS imaging sensor. A microlens array within the SHWS samples the incident wavefront, generating a pattern of focal spots on a CCD. Real-time centroid analysis of these spots provides a map of local wavefront slopes. These measurements are processed through a singular value decomposition (SVD) algorithm to fit a Zernike polynomial basis set, enabling real-time reconstruction of the wavefront phase. The defocus component (S) is extracted from the second-order Zernike coefficients, providing a direct, quantitative measure of the refractive error in diopters. This value serves as a precise error signal in a closed-loop control system, which commands a voice-coil actuated focusing lens to its null position in a single, deterministic step, eliminating the need for iterative search algorithms. ResultsComprehensive evaluation demonstrated the system’s high performance. Testing on a calibrated model eye (OEMI-7) established a highly linear relationship between the computed defocus S and the focusing lens position across a ±20 Diopter (D) compensation range, achievable within a 5 mm mechanical travel. The system achieved a focusing precision of 0.08 D, corresponding to an 18-fold improvement over a conventional projection spot-size method tested under identical conditions. The total focus acquisition time, encompassing wavefront measurement, computation, and lens actuation, averaged under 0.5 s. Clinical validation with 25 human volunteers (50 eyes, refractive range -15 D to +10 D) confirmed practical efficacy. The wavefront-sensing AF succeeded in 92% of attempts with a mean time of 0.5 s, substantially outperforming a projection-based benchmark which achieved only a 32% success rate with an average time of 4.25 s. The system provided instantaneous directional guidance and maintained stability during minor ocular movements. Objective assessment of image quality, via amplitude contrast of retinal vasculature, showed consistent and significant enhancement following AF correction across the entire tested diopter range. ConclusionThis work successfully implements and validates a direct wavefront-sensing autofocus paradigm for portable fundus cameras. By directly quantifying and compensating for the optical defocus aberration, this method bypasses the fundamental limitations of image-processing and projection-based techniques, enabling rapid, precise, and deterministic diopter compensation. The developed system delivers an exceptional combination of a wide operational range (±20 D), high accuracy (0.08 D), fast convergence (0.5 s), and a compact physical footprint. This technology provides a practical and high-performance focusing solution capable of enhancing the reliability, throughput, and diagnostic utility of portable retinal imaging in large-scale screening applications. Future efforts will be directed towards system cost optimization and performance adaptation for diverse ocular conditions.
4.Evaluation of CARIFS Score and Negative Antigen Conversion Rate of Qingxuan Daozhi Formula in Treatment of Influenza in Children (Heat Accumulation in Lung and Stomach Syndrome):A Multi-center Randomized Controlled Clinical Study
Jing WANG ; Liqun WU ; Tiegang LIU ; Yongning CAO ; Jing QIU ; Jing LI ; Huaqing TAN ; Ying ZHANG ; Xulei GOU ; Jia WANG ; Jing LI ; Haipeng CHEN ; Xueying QIN ; Yuanshuo TIAN ; Yang WANG ; Chen BAI ; Zhendong WANG ; Qianqian LI ; He YU ; Xueyan MA ; Fei DONG ; Lin JIANG ; Yingqi XU ; Jianping LIU ; Xiaohong GU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):188-196
ObjectiveThis paper aims to observe the syndrome improvement and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children (heat accumulation in the lung and stomach syndrome). MethodsThrough a multi-center randomized controlled methodology design,confirmed influenza cases were collected from October 2022 to April 2023 in the pediatrics department of eight hospitals,such as Dongfang Hospital of Beijing University of Chinese Medicine. A total of 180 children with influenza and heat accumulation in the lung and stomach syndrome conforming to the standard were recruited through the clinic. The sick children meeting the inclusion criteria were randomly divided into groups by a block-randomized method. The children in the experimental group were treated with Qingxuan Daozhi formula for five days,and those in the control group were treated with Oseltamivir Phosphate Granules for five days. The primary efficacy indicator was the negative conversion rate of influenza antigen detection. Secondary efficacy indicators were the Canadian acute respiratory illness and flu scale (CARIFS) and the incidence of complications,severe cases, and critical cases. Follow-up observation was conducted on the day of enrollment,48 hours after medication,72 hours after medication, and (6+1) d after medication. ResultsOne hundred and eighty participants were randomly assigned to the experimental group (90 cases) or the control group (90 cases). All participants were followed up during the study. Comparison of influenza antigen detection results in the primary efficacy indicators showed that the average time of negative influenza antigen conversion in the experimental group was (5.29±1.25) d,and that in the control group was (5.40±1.68) d,without a statistically significant difference. After five days of intervention,52 cases in the experimental group and 51 cases in the control group converted to negative,without a statistically significant difference. CARIFS score results in the secondary efficacy indicators showed that during 72 hours after intervention,there were statistically significant differences between the experimental group and the control group in three dimensions, including headache,muscle soreness, and the need for extra care (P<0.05). On the (6+1) days after the intervention,the differences in both the experimental group and the control group were statistically significant in 10 dimensions, including sore throat,bad sleep,uncomfortable feeling,poor spirit and fatigue,crying more than usual,the need for extra care,symptom,function,influence on parents,and total score (P<0.05). The comparison results within the group in the dimensional scores of symptom, function, and influence on parents,as well as the CARIFS total score showed that with the delay of follow-up time,scores of both groups decreased significantly,with a statistically significant difference (P<0.01). Inter-group comparison results showed that the mean score of the experimental group was higher than that of the control group at the time of enrollment. With the progress of intervention,the score of the experimental group was significantly decreased compared with that of the control group. At the end of follow-up,the mean score of the experimental group was lower than that of the control group,with no statistically significant difference. In terms of the incidence of complications,severe cases, and critical cases, there were no complications,severe cases, and critical cases in the two groups,without a statistically significant difference. ConclusionThe symptom improvement effect and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children (heat accumulation in the lung and stomach syndrome) are not inferior to Oseltamivir Phosphate granules, and children's acceptance is better. It can be more widely used in clinical treatment of influenza in children (heat accumulation in the lung and stomach syndrome).
5.Multidimensional analysis of concurrent proximal bronchiolar adenoma and lung carcinoma
Lu-Yao LI ; Gong-Ming DONG ; Yun-Peng ZHANG ; Ting-Ting WANG ; Fu-Quan JIA ; Guan-Jun ZHANG
Journal of Pathology and Translational Medicine 2026;60(3):356-363
Bronchiolar adenoma (BA) is a rare type of lung tumor characterized by bilayered epithelial cells having a continuous basal layer and a luminal layer. It resembles mucinous adenocarcinoma (MA) on frozen section, with difficulty in distinguishing the basal layer. Immunohistochemistry is the best choice for verifying the diagnosis. This study aimed to comprehensively characterize three cases of BA-combined carcinoma using clinical, histopathological, and genetic features. BA and carcinoma sections were subjected to next-generation sequencing, respectively. It was hypothesized that while different mutation forms matched different regions, BA and lung adenocarcinoma shared the same gene mutation when they co-occurred in the same location. BA with extensive carcinoma is extremely rare and presents diagnostic challenges due to its overlap with conditions such as MA. Because of its distinctive morphological characteristics, BA may be regarded as a low-grade malignancy, particularly during a confusing evaluation. A multifaceted examination of clinical, radiological, immunohistochemical, and genetic data is necessary for an accurate diagnosis.
6.Association study of SLC7A11 gene polymorphisms with helicobacter pylori infection and non-cardia gastric cancer
Linli WANG ; Fang GAO ; Wenjie DONG ; Ning CHU ; Yanbin JIA
Acta Universitatis Medicinalis Anhui 2026;61(6):1124-1129
ObjectiveTo explore the correlation between the polymorphisms of the solute carrier family7 member11 (SLC7A11) gene and the risk of non-cardia gastric cancer (NCGC) as well as the infection of Helicobacter pylori (H.pylori). MethodsA total of 435 patients with NCGC and 479 healthy individuals undergoing physical examinations were included. The polymorphisms of the SLC7A11 gene (rs7674870, rs11734488, rs12506138, rs10022036) were genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The influence of each locus on H. pylori infection and the occurrence of NCGC was analyzed. ResultsNo significant associations were found among SNP rs7674870, rs11734488, and rs12506138 and H. pylori infection (P>0.05). The CT genotype and CC genotype of rs12506138, when in a codominant model, reduced the risk of developing NCGC compared to the TT genotype; the A-C-C and A-T-C haplotypes constructed by SLC7A11 rs7674870, rs11734488, rs12506138 were associated with a reduced risk of NCGC; the A-T-T and G-C-C haplotypes were associated with an increased risk of NCGC. ConclusionSLC7A11 rs7674870 and rs11734488 are not pivotal genetic loci for H. pylori infection and the pathogenesis of NCGC. SLC7A11 rs12506138 acts as a protective genetic locus for NCGC and antagonizes its oncogenic progression. The haplotype composed of the above three loci participates in regulating the overall genetic susceptibility to NCGC.
7.The epidemiologic characteristics of parasitic and vector-borne diseases in Minhang and Jinshan districts, Shanghai
Dun-jia WANG ; Xiao-lu HE ; Tian-tian ZHANG ; Tang WANG ; Zhao-peng DONG ; Can-lei SONG ; Shi-ji TAO ; Zheng-xing HOU ; Yu-ping CHENG ; Yi-bin ZHOU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(3):173-179
Objective The aim of the study is to analyze the incidence patterns and epidemiological characteristics of parasitic and vector-borne diseases in the Minhang and Jinshan districts of Shanghai, and provide evidence-based information for optimizing prevention and control measures. Methods We collected information from epidemic reports and epidemiological investigation forms for cases of parasitic and vector-borne diseases from 2005 to 2024 in Shanghai's Minhang and Jinshan districts. Descriptive epidemiological method were adopted to conduct a comparative analysis of incidence trends, epidemic characteristics, and infection sources. Results A total of 229 cases in 7 categories were reported between 2005 and 2024, with an average annual incidence rate of 0.36/100 000. The average annual incidence rate in Minhang District was significantly higher than that in Jinshan District. The most prevalent diseases were malaria and dengue fever, followed by Japanese encephalitis. Monthly case reports were available, predominantly from June to October, with a peak in July-August. A statistically significant difference was observed between the two districts in case distribution during June-October (P<0.001), whereas no significant difference was found for July-August (P=0.331). The majority of cases were young men (111/229,48.47%), aged 20-39 years. Migrant workers/farmers constituted the largest occupational group (55/229, 24.02%). Registered residents of other provinces and cities (147/229,64.19%), and the main source of infection was overseas importation (112/229,48.91%). There were no statistically significant differences in gender and household-registration composition between the two districts (P>0.05). Significant differences existed in age, occupation, and source of infection (P<0.05). The imported diseases were malaria and dengue fever, with Africa and Southeast Asia being the primary source locations. Malaria was the predominant disease imported from other provinces and cities, with cases having been imported from abroad since 2012. Local infections were mainly Japanese encephalitis, followed by malaria (with the last local case reported in 2010). Cases between 2005 and 2009 were predominantly imported from other provinces and cities. The proportion of cases imported from abroad increased sharply after 2010, particularly in Minhang District. Local infections sometimes arise in Jinshan District, with Japanese encephalitis serving as the primary type. Conclusions Incidence rates of parasitic and vector-borne diseases in Minhang and Jinshan districts have generally remained low. To further reduce this disease burden, it will be necessary to strengthen joint measures for prevention and control, eliminate the environmental risk factors of vector organisms, enhance monitoring and precise control of the environment and inbound personnel to prevent the occurrence and transmission of parasitic and vector-borne diseases.
8.Role of Adipose Tissue Macrophages and Adipokine in The Pathogenesis of Obesity-related Immune Diseases
Zi-Xuan JIA ; Qing-Lu WANG ; Pan-Pan DONG ; Xiao-Tong YUAN
Progress in Biochemistry and Biophysics 2026;53(10):2537-2557
Adipose tissue macrophages (ATMs) are crucial immunomodulatory factors in the adipose tissue (AT) microenvironment, playing an irreplaceable role in maintaining the balance of the local immune system and regulating metabolic homeostasis. Under obese conditions, the excessive accumulation of lipids leads to abnormal expansion of adipose tissue, which further disrupts the homeostasis of the local microenvironment, including the imbalance of inflammatory factors, the occurrence of oxidative stress, and the damage of microcirculation. As an essential immune cell population in adipose tissue, ATMs are deeply involved in the occurrence and progression of metabolic disorders and multiple obesity-related diseases, and their functional abnormalities are closely related to the initiation and development of adipose tissue inflammation and systemic metabolic disorders. The interaction between ATMs and adipokines, including adiponectin, leptin, resistin and retinol-binding protein 4 (RBP4), acts as the core immunological and molecular mechanism linking adipose tissue inflammation to continuous disease deterioration, and this interaction is also a key research focus in the field of obesity-related diseases in recent years. This review discusses in detail the facilitating effect of the AT inflammatory microenvironment on the differentiation of peripheral monocytes into ATMs, elaborating on the specific molecular mechanisms by which various inflammatory mediators and abnormal metabolic products in the inflammatory microenvironment induce the differentiation of peripheral monocytes into functional ATMs. It also focuses on the regulatory mechanisms of chemokine-mediated ATM polarization and recruitment, including the specific roles of key chemokines like monocyte chemoattractant protein‑1 (MCP-1) and C‑X‑C motif chemokine ligand 10 (CXCL10) in mediating the recruitment of ATMs to adipose tissue, as well as the molecular pathways that regulate the switch between M1 pro-inflammatory phenotype and M2 anti-inflammatory phenotype of ATMs. In addition, this review explores the specific process by which ATMs trigger AT inflammation by secreting various pro-inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β, which further amplify the local inflammatory response and disrupt the metabolic homeostasis of adipose tissue. Furthermore, it analyzes the specific binding patterns and molecular characteristics of adiponectin, leptin, resistin and RBP4 with ATM surface receptors, and clarifies how the activation of downstream immune signaling pathways, such as Janus kinase‑signal transducer and activator of transcription (JAK-STAT), nuclear factor‑κB (NF‑κB), and phosphatidylinositol 3‑kinase‑protein kinase B (PI3K-AKT), triggered by these binding processes, induces the occurrence and development of various obesity-related diseases, including insulin resistance, cancer, metabolic-associated steatohepatitis (MASH) and cardiovascular diseases. This paper highlights the unique and crucial role of the interaction between ATMs and adipokines in the occurrence and progression of obesity-related diseases, emphasizing that this interaction is a key link connecting local adipose tissue inflammation to systemic metabolic disorders. It further summarizes the potential therapeutic strategies targeting the ATM-adipokine axis, and comprehensively illustrates three major clinical application barriers, namely biological complexity, technical bottlenecks, and clinical translation obstacles, in the process of applying these therapeutic strategies. For each barrier, this review puts forward corresponding feasible solutions and clear research directions, which not only enrich the theoretical system of the ATM-adipokine axis in the field of obesity research, but also provide a solid theoretical basis and practical ideas for the clinical intervention, prevention and treatment of obesity and its related diseases.
9.Gantou Fumu Decoction Inhibits Hepatic Stellate Cell Activation by Regulating Nrf2/ARE Signaling Pathway
Wangsheng HE ; Yanping ZHU ; Shizhen LONG ; Mengmeng LI ; Wenming YANG ; Han WANG ; Meixia WANG ; Ting DONG ; Shupei JIA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(23):105-115
ObjectiveTo explore the mechanism through which Gantou Fumu decoction (GDFMT)-containing serum inhibits the activation of hepatic stellate cells. MethodsSerum pharmacology techniques were employed to prepare GDFMT-containing serum samples. Rat hepatic stellate cells (HSCT6) were randomly allocated into three groups: Control, penicillamine (90 mg·kg-1), and GDFMT (6.8 g·kg-1). Cell viability was assessed by the cell counting kti-8 (CCK-8) assay, intracellular ROS levels were determined by flow cytometry using the DCFH‑DA fluorescent probe and while extracellular collagen Ⅰ (ColⅠ) levels were quantified via enzyme linked immunosorbent assay (ELISA). Intracellular α-smooth muscle actin (α-SMA) expression was visualized through immunofluorescence staining. Real-time polymerase chain reaction(Real-time PCR) and Western blot were performed to evaluate alterations in the expression of nuclear factor-erythroid 2-related factor 2 (Nrf2), glutamate cysteine ligase, catalytic (GCLC), glutamate-cysteine ligase, modifier subunit (GCLM), Kelch-like ECH-associated protein 1 (Keap1), NAD(P)H:Quinone oxidoreductase 1 (NQO1), and heme oxygenase-1 (HO-1). Genetic knockdown of Nrf2 was achieved through small interfering (siRNA) transfection to validate pathway specificity. ResultsCCK-8 data indicated that under the selected experimental conditions, culturing HSCT6 cells with 10% (volume fraction) GDFMT-containing serum for 48 h demonstrated equivalent effect in inhibiting HSCT6 cell proliferation compared with culturing HSCT6 cells with 10% (volume fraction) penicillamine-containing serum for 36 h, with no statistically significant difference between the two groups. After treatment of HSCT6 cells with siRNA-mediated Nrf2 gene transfection technique, the inhibitory effect of GDFMT on the activation process of HSCT6 cells was significantly weakened. After HSCT6 cells were cultured with GDFMT-containing serum for 48 h, the flow cytometry with a fluorescence probe showed that the content of reactive oxygen species (ROS) in the cells was significantly reduced. After the expression level of the Nrf2 gene in HSCT6 cells was knocked down by siRNA transfection, the content of ROS increased (P<0.01), indicating that GDFMT significantly reduced the ROS level in activated HSCT6 cells, and this effect strictly depended on the integrity of the Nrf2 pathway. Quantitative analysis by ELISA showed that the synthesis of ColⅠ in the HSCT6 cell culture system treated with GDFMT-containing serum for 48 h was lower than that in the control group (P<0.01). After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection, the ColⅠ concentration in the culture supernatant increased (P<0.01). The results of α-SMA detection by IF method showed that compared with the normal group, GDFMT-containing serum reduced the expression of α-SMA (P<0.01) and the protein level of α-SMA in HSCT6 cells was increased (P<0.01) after siRNA-mediated Nrf2 silencing, suggesting that Nrf2 signaling pathway may be involved in regulating the phenotypic transformation process of the cells. Real-time PCR and Western blot results showed that GDFMT-containing serum reduced the protein level of α-SMA (P<0.01) and enhanced the gene and protein levels of Nrf2 in the nucleus and GCLC, HO-1, Keap1, GCLM, and NQO1 in the cytoplasm of HSCT6 cells (P<0.05, P<0.01). After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection, the protein level of α-SMA was increased (P<0.01), and the levels of GCLC, GCLM, Keap1, NQO1, and HO-1 were decreased (P<0.05, P<0.01). These results indicated that GDFMT-containing serum successfully triggered the Nrf2 signaling pathway of HSCT6 cells. ConclusionThe anti-hepatic fibrosis effect of 10%GDFMT was equivalent to that of penicillamine. In addition, GDFMT can inhibit HSC activation and reduce oxidative stress damage by activating the Nrf2/antioxidant response element (ARE) pathway, as manifested by the up-regulated expression of Nrf2 and downstream antioxidant genes (GCLC, GCLM, HO-1, Keap1, and NQO1) and reduced secretion of ROS, α-SMA, and ColⅠ.
10.Development of an Analytical Software for Forensic Proteomic SAP Typing
Feng HU ; Meng-Jiao WANG ; Jia-Lei WU ; Dong-Sheng DING ; Zhi-Yuan YANG ; An-Quan JI ; Lei FENG ; Jian YE
Progress in Biochemistry and Biophysics 2025;52(9):2406-2416
ObjectiveThe proteome of biological evidence contains rich genetic information, namely single amino acid polymorphisms (SAPs) in protein sequences. However, due to the lack of efficient and convenient analysis tools, the application of SAP in public security still faces many challenges. This paper aims to meet the application requirements of SAP analysis for forensic biological evidence’s proteome data. MethodsThe software is divided into three modules. First, based on a built-in database of common non-synonymous single nucleotide polymorphisms (nsSNPs) and SAPs in East Asian populations, the software integrates and annotates newly identified exonic nsSNPs as SAPs, thereby constructing a customized SAP protein sequence database. It then utilizes a pre-installed search engine—either pFind or MaxQuant—to perform analysis and output SAP typing results, identifying both reference and variant types, along with their corresponding imputed nsSNPs. Finally, SAPTyper compares the proteome-based typing results with the individual’s exome-derived nsSNP profile and outputs the comparison report. ResultsSAPTyper accepts proteomic DDA mass spectrometry raw data (DDA acquisition mode) and exome sequencing results of nsSNPs as input and outputs the report of SAPs result. The pFind and Maxquant search engines were used to test the proteome data of 2 hair shafts of2 individuals, and both obtained SAP results. It was found that the results of the Maxquant search engine were slightly less than those of pFind. This result shows that SAPTyper can achieve SAP fingding function. Moreover, the pFind search engine was used to test the proteome data of 3 hair shafts from 1 European person and 1 African person in the literature. Among the sites fully matched by the literature method, sites detected by SAPTyper are also included; for semi-matching sites, that is, nsSNPs are heterozygous, both literature method and SAPTyper method had the risk of missing detection for one type of the allele. Comparing the analysis results of SAPTyper with the SAP test results reported in the literature, it was found that some imputed nsSNP sites identified by the literature method but not detected by SAPTyper had a MAF of less than 0.1% in East Asian populations, and therefore they were not included in the common nsSNP database of East Asian populations constructed by this software. Since the database construction of this software is based on the genetic variation information of East Asian populations, it is currently unable to effectively identify representative unique common variation sites in European or African populations, but it can still identify SAP sites shared by these populations and East Asian populations. ConclusionAn automated SAP analysis algorithm was developed for East Asian populations, and the software named SAPTyper was developed. This software provides a convenient and efficient analysis tool for the research and application of forensic proteomic SAP and has important application prospects in individual identification and phenotypic inference based on SAP.


Result Analysis
Print
Save
E-mail