1.Xiaozheng Zhitong Paste Alleviates Bone Cancer Pain by Regulating PD-1/PD-L1-induced Osteoclast Formation
Lu SHANG ; Juanxia REN ; Guangda ZHENG ; Linghan MENG ; Lingyun WANG ; Changlin LI ; Dongtao LI ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):72-79
ObjectiveThis study aims to investigate the action mechanism by which Xiaozheng Zhitong paste (XZP) alleviates bone cancer pain (BCP) by regulating programmed death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) pathway-induced osteoclast formation. MethodsThirty female C57BL/6 mice were randomly allocated into the following groups (n=6 per group): normal control group, model group, low‑dose XZP group (31.5 g·kg-1), high‑dose XZP group (63 g·kg-1), and PD‑1 inhibitor (Niv) group. A bone cancer pain (BCP) model was established by injecting Lewis lung carcinoma cells. Mice in the normal control and model groups received topical application of a blank paste matrix at the wound site. Mice in the low‑ and high‑dose XZP groups were treated with XZP applied topically twice daily. Mice in the Niv group were topically administered the blank paste matrix and additionally received Niv via tail‑vein injection every two days. All interventions were continued for 21 days. During this period, behavioral tests were performed to assess mechanical, motor, and thermal nociceptive sensitivities. After 21 days, all mice were euthanized, and bone tissue from the operated side was collected for sectioning and preservation. Tartrate‑resistant acid phosphatase (TRAP) staining was used to evaluate osteoclast expression in the lesioned bone tissue. Immunohistochemistry was performed to detect the expression of Runt‑related transcription factor 2 (Runx2) in the lesioned bone tissue. Immunofluorescence was employed to assess the expression of PD‑1 and PD‑L1 in the lesioned bone tissue. ResultsCompared with the normal group, the model group showed significantly decreased limb mechanical withdrawal threshold, spontaneous paw flinching, and thermal withdrawal latency (P<0.01), increased number of osteoclasts in the lesioned bone tissue (P<0.01), and reduced expression of Runx2 (P<0.01). Compared with the model group, the BCP mice in the XZP low-dose group, XZP high-dose group, and Niv group exhibited increased limb mechanical withdrawal threshold, movement scores, and thermal withdrawal latency (P<0.01). The XZP low-dose group showed no significant changes in osteoclast number or Runx2 expression, while the XZP high-dose group and Niv group demonstrated significantly reduced osteoclast numbers (P<0.01) and significantly increased Runx2 expression (P<0.01). In the lesioned bone tissue of BCP mice, the XZP low-dose group showed no significant decrease in the percentage of PD-1 expression, but a decrease in the percentage of PD-L1 expression (P<0.05). In contrast, both the XZP high-dose group and the Niv group exhibited significant reductions in the percentages of PD-1 and PD-L1 expression (P<0.01). ConclusionXZP alleviates the pain of mice with BCP by blocking the PD-1/PD-L1 pathway to inhibit osteoclastogenesis.
2.Xiaozheng Zhitong Paste Relieves Bone Cancer Pain in Mice by Alleviating Activation of Microglia in Spinal Cord and Damage to Neurons via Blocking PAR2/NF-κB/NLRP3 Pathway
Guangda ZHENG ; Linghan MENG ; Lu SHANG ; Juanxia REN ; Dongtao LI ; Haixiao LIU ; Lingyun WANG ; Changlin LI ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):91-100
ObjectiveTo investigate the effects and underlying mechanisms of Xiaozheng Zhitong Paste (XZP) on bone cancer pain (BCP). MethodsThirty female BALB/c mice were randomly divided into five groups: a Sham group, a BCP group, a BCP+low-dose XZP group, a BCP+high-dose XZP group, and a BCP+high-dose XZP + protease-activated receptor 2 (PAR2) agonist GB-110 group. BCP mice model was constructed by injecting Lewis lung carcinoma cells into the femoral cavity of the right leg, which was followed by being treated with XZP for 21 d. After 21 d, the mice were sacrificed. Nissl staining was used to evaluate the survival of spinal cord neurons. Immunofluorescence staining was conducted to localize ionized calcium-binding adapter molecule 1 (Iba1) and neuronal nuclear antigen (NeuN) in spinal cord tissue, thereby assessing microglial activation and neuronal survival. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), transforming growth factor-β (TGF-β), interleukin-4 (IL-4), and interleukin-10 (IL-10) in spinal cord tissue. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression levels associated with M1/M2 polarization of microglia. Western blot analysis was performed to examine the expression of proteins related to microglial polarization as well as those involved in the PAR2/nuclear factor kappa B (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway in the spinal cord. ResultsCompared with the Sham group, the spinal cord neurons were damaged, the number of Nissl-positive spinal cord neurons in the spinal cord tissue was significantly reduced (P<0.01), and the rate of NeuN-positive cells was significantly decreased (P<0.01). The spinal cord microglia were activated, the inflammatory level of the spinal cord tissue was enhanced, and Iba1 staining was significantly enhanced (P<0.01). The levels of IL-1β, TNF-α, IL-6, TGF-β, IL-4 and IL-10 were significantly increased (P<0.01). The mRNA expressions of IL-1β, TNF-α and inducible nitric oxide synthase (iNOS) were significantly increased (P<0.01), and the expression of PAR2, NLRP3, ASC and NF-κB p65 proteins in the spinal cord tissue of the BCP mice was significantly enhanced (P<0.01). Compared with the BCP group, high-dose XZP treatment significantly increased the number of Nissl-positive spinal cord neurons in the BCP mice (P<0.01), significantly enhanced the rate of NeuN-positive cells in the spinal cord tissue, and significantly weakened Iba1 staining (P<0.01). In addition, the levels of IL-1β, TNF-α, and IL-6 were significantly decreased, while the levels of TGF-β, IL-4, and IL-10 were significantly increased (P<0.05, P<0.01). The mRNA expression levels of IL-1β, TNF-α, and iNOS were decreased, whereas those of cluster of differentiation 206 (CD206), arginase-1 (Arg-1), and YM1/2 were significantly increased (P<0.05, P<0.01). Low-dose and high-dose XZP treatment significantly decreased the expression of PAR2, NLRP3, ASC, and NF-κB p65 proteins in the spinal cord tissue (P<0.05, P<0.01). These effects could all be significantly eliminated by the PAR2 agonist GB-110. ConclusionXZP can mitigate BCP in mice, which may be achieved through blocking the activated PAR2/NF-κB/NLRP3 pathway.
3.Xiaozheng Zhitong Paste Alleviates Bone Cancer Pain of Mice by Reducing Ferroptosis in Spinal Cord Tissue and Neuronal Damage via Regulating Nrf2/HO-1/GPX4/SLC7A11 Signaling Pathway
Juanxia REN ; Lu SHANG ; Guangda ZHENG ; Linghan MENG ; Lingyun WANG ; Changlin LI ; Dongtao LI ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):101-113
ObjectiveThe paper aims to investigate the action mechanism by which the Xiaozheng Zhitong paste (XZP) relieves bone cancer pain (BCP). MethodsA model of mice with BCP was established by using Lewis tumor cells. The therapeutic effects of XZP, the ferroptosis inhibitor Ferrostatin-1 (Fer-1), and the nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor Brusatol (Bru) on BCP were examined. Mice were randomly divided into the Sham operation group, BCP group, BCP+XZP-L group, BCP+XZP-H group, BCP+Fer-1 group, and BCP+XZP-H+Bru group, with six mice in each group. Pain behavior tests were conducted on the mice to assess pain levels. Colorimetric assays were employed to measure ferroptosis-related factors in serum and spinal cord tissue including Fe, malondialdehyde (MDA), reactive oxygen species (ROS), and superoxide dismutase (SOD). Immunofluorescence staining was used to assess ROS production in spinal cord tissue. Transmission electron microscopy was used to observe the ultrastructure of mitochondria in lumbar spinal cord tissue. Quantitative real-time polymerase chain reaction (Real-time PCR) was employed to detect mRNA expression of Nrf2, heme oxygenase-1 (HO-1), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) in spinal cord neuron tissue. The protein expression of Nrf2, HO-1, GPX4, and SLC7A11 in spinal cord neurons was measured by Western blot. ResultsCompared with the Sham group, mice in the BCP group exhibited significantly reduced limb usage scores, mechanical foot withdrawal thresholds, and thermal foot withdrawal thresholds (P<0.01). Serum and lumbar spinal cord tissue levels of Fe, MDA, and reactive oxygen species (ROS) were significantly elevated (P<0.05), while superoxide dismutase (SOD) levels were significantly decreased (P<0.05). Lumbar spinal cord mitochondrial structural damage was observed, and mRNA and protein expression of Nrf2, HO-1, GPX4, and SLC7A11 were significantly downregulated (P<0.01). Compared with the BCP group, both low- and high-dose XZP groups improved the aforementioned pain behavioral indicators (P<0.05,P<0.01), reduced ferroptosis-related biomarkers including Fe, MDA, and ROS levels (P<0.05), increased SOD levels (P<0.05,P<0.01), alleviated mitochondrial damage, and upregulated Nrf2, HO-1, GPX4, SLC7A11 mRNA and protein expression (P<0.05,P<0.01). The high-dose XZP group exhibited comparable efficacy to Fer-1 in alleviating pain and inhibiting ferroptosis. Following Bru administration, XZP's effects on pain behavioral indicators, regulation of ferroptosis-related markers, mitochondrial structural protection, and activation of the Nrf2/HO-1/GPX4/SLC7A11 pathway were significantly reversed (P<0.05,P<0.01). ConclusionExternal application of XZP alleviates pain symptoms in BCP mice by activating the Nrf2/HO-1/GPX4/SLC7A11 pathway, thereby inhibiting ferroptosis and neuronal damage in spinal cord neurons.
4.Discussion on Treatment of Cancer Pain with Modified Wumeiwan Based on Jueyin Syndrome
Haixiao LIU ; Linghan MENG ; Guangda ZHENG ; Dongtao LI ; Lu SHANG ; Juanxia REN ; Changlin LI ; Lingyun WANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):124-128
Pain, as one of the most common symptoms in cancer patients, seriously affects the survival quality of patients. The three-step pain relief program currently used in clinical practice cannot completely relieve pain in cancer patients and is accompanied by many problems. From the perspective of Jueyin syndrome in traditional Chinese medicine (TCM), this paper believed that the core pathogenesis of cancer pain was declined healthy Qi and cold and heat in complexity, and used Wumeiwan as the main formula with modification according to syndrome for clearing the upper, warming the lower part of the body, and harmonizing the cold and heat. It can regulate the pathological environment of deficiency, cold, stasis, toxicity, and heat, and restore the physiological function of Yang transforming Qi while Yin constituting form, so as to prevent, relieve, and even eliminate cancer pain, having achieved good clinical efficacy. It can not only help cancer patients relieve pain, but also control tumor and eliminate tumor, achieving a dual benefit of pain relief and tumor suppression. It gives full play to the characteristics and advantages of syndrome differentiation and treatment in TCM, and expands the scope of ZHANG Zhongjing's treatment for Jueyin syndrome, which provides ideas for the clinical diagnosis and treatment of cancer pain from the perspective of deficiency-excess in complexity and cold and heat in complexity.
5.Spatiotemporal distribution characteristics of and trends in disease burden of dengue fever in China from 2005 to 2024
Lianfang FENG ; Meng SHANG ; Jiarong REN ; Xiaoxu WANG ; Haoqiang JI ; Xinning HAO ; Jing LI ; Qiyong LIU
Chinese Journal of Schistosomiasis Control 2026;38(2):137-147
Objective To analyze the spatiotemporal distribution characteristics of and trends in the disease burden of dengue fever in China from 2005 to 2024, so as to provide insights into formulation of dengue fever control strategies. Methods Data pertaining to dengue fever cases in China from 2005 to 2024 were retrieved from the Infectious Disease Reporting Information System of Chinese Center for Disease Control and Prevention, and city population, gross domestic product (GDP), GDP per capita, and consumer price index in China were captured from the China Statistical Yearbook, National Bureau of Statistics of China, the China City Statistical Yearbook, and bureaus of statistics in each city. The disability-adjusted life years (DALYs), years of life lost (YLLs), and years lived with disability (YLDs) due to dengue fever were calculated in China from 2005 to 2024. The direct and indirect economic burdens of dengue fever were estimated to calculate the total economic burden. The trends in the disease burden of dengue fever were estimated in China from 2005 to 2024 using a Joinpoint regression model with the software Joinpoint 4.9.0.0, and the average annual percent change (AAPC) and its 95% confidence interval (CI) were calculated. In addition, the DALYs rate and economic burden of dengue fever in China were subjected to global and local spatial autocorrelation analyses using the software ArcGIS 10.8. Results The gross DALYs due to dengue fever were 5 558 person-years in China from 2005 to 2024, and the DALYs of dengue fever increased from 36 person-years in 2005 to 899 person-years in 2024, with an increase of 23.97 folds. The average annual DALYs rate of dengue fever was 0.02 person-years/105 in China during the 20-year study period from 2005 to 2024, and the DALYs rate peaked in 2014 (0.13 person-years/105) and reduced during the COVID-19 pandemic from 2020 to 2022. YLDs were the main contributor of DALYs due to dengue fever in China from 2005 to 2024, with a total of 5 354 person-years, accounting for 96.33% (5 354 person-years/5 558 person-years) of the gross DALYs. The gross DALYs of dengue fever were 2 982 person-years among men (53.66%) and 2 575 person-years among women (46.34%) in China from 2005 to 2024, and high DALYs of dengue fever were measured among residents at ages of 15 to 30 years (1 639 person-years), 30 to 45 years (1 857 person-years), and 45 to 60 years (1 204 person-years), respectively, accounting for 84.56% (4 700 person-years/5 558 person-years) of total DALYs due to dengue fever in China. The total economic burden of dengue fever was estimated to be 612 million Yuan in China from 2005 to 2024, with an average annual economic burden of 30.584 million Yuan. The economic burden of dengue fever increased from 196 000 Yuan in 2005 to 121 million Yuan in 2024 in China, with an increase of 616.35 folds, and the per capita economic burden increased from 3 322.21 Yuan in 2005 to 4 940.01 Yuan in 2024, with an increase of 48.70%. Dengue fever cases were reported in 274 cities (counties) across 31 provinces (autonomous regions, municipalities) in China from 2005 to 2024, with relatively higher DALYs in Guangdong Province and Yunnan Province. Spatial autocorrelation analysis revealed that the disease burden of dengue fever appeared positive aggregation in Chinese cities (counties) from 2005 to 2024 (global Moran’s I = 0.045, Z = 2.24, P < 0.05), with high-high clusters mainly concentrated in the Pearl River Delta region in Guangdong Province and Xishuangbanna Dai Autonomous Prefecture and Pu’er City in Yunnan Province, and the total economic burden (global Moran’s I = 0.032, Z = 9.55, P < 0.001), per capita economic burden (global Moran’s I = 0.208, Z = 27.34, P < 0.001), and the proportion of total economic burdens in GDP in 2024 (global Moran’s I = 0.017, Z = 5.91, P < 0.001) all presented positive aggregation, with relatively higher total economic burdens mainly concentrated in Guangdong Province and Yunnan Province. Joinpoint regression analysis showed that the gross DALYs rates of dengue fever appeared an overall tendency towards a rise in China from 2005 to 2024 (AAPC = 16.24%, P = 0.029), and the DALYs rate presented an overall tendency towards a rise among both men (AAPC = 14.75%, P = 0.028) and women (AAPC = 14.93%, P = 0.037) during the study period. The per capita direct economic burden appeared an overall tendency towards a rise among dengue fever patients in China from 2005 to 2024 (AAPC = 2.16%, P = 0.012); however, there was no significant difference in the trends in the per capita indirect economic burden (AAPC = 0.46%, P = 0.470). In addition, the DALYs rate of dengue fever appeared a tendency towards a rise in 84.67% (232/274) of cities (counties) in China from 2005 to 2024, and the per capita economic burden appeared a tendency towards a rise in 85.40% (234/274) of cities (counties), while the DALYs rate and per capita economic burden of dengue fever appeared a tendency towards a rise in 77.01% (211/274) of cities (counties). Conclusions The disease burden of dengue fever significantly increased in China from 2005 to 2024. It is recommended to reinforce integrated dengue fever control in high-risk areas and among high-risk populations, and to improve the surveillance of imported dengue fever cases and vector control.
6.Raman Enhancement-Nanozyme Dual Functional Nanocomposite Au@MnO2-based Surface Enhanced Raman Scattering Biosensor for Organophosphorus Pesticides
Xiao-Tong ZHANG ; Meng-Ren XUAN ; Fu-Gang XU ; Yue ZHANG
Chinese Journal of Analytical Chemistry 2025;53(10):1664-1673,中插1-中插4
Surface enhanced Raman scattering(SERS)is an important technique for detecting pesticide residues.However,many pesticides have small Raman scattering cross sections and low affinity for gold/silver enhancement substrates,resulting in low sensitivity for direct SERS detection.Indirect methods typically require complex surface modifications and have poor universality.Here,a dual functional composite Au@MnO2 combined with acetylcholinesterase(AChE)was proposed for SERS biosensing of pesticides.The Au@MnO2 with oxidase-like activity could oxidize 3,3′,5,5′-tetramethylbenzidine(TMB)into oxTMB with a large Raman scattering cross section,producing strong SERS signal using Au@MnO2 itself as enhancement substrate.In the presence of AChE,acetylthiocholine(ATCh)was catalytically hydrolyzed to produce thiocholine(TCh),and its reducibility could induce the decomposition of MnO2.Consequently,oxidase-like activity of Au@MnO2 was declined,less oxTMB was produced,and gold nanoparticles(AuNPs)detached from the MnO2 layer,resulting in a weakened SERS response.Organophosphorus pesticides(OPs),as inhibitors of AChE,prevented the production of TCh,thereby blocking the decomposition of MnO2 and leading to the recovery of Raman signals.Under optimized conditions,the Raman signal switching strategy was successfully used for detection of three OPs,i.e.,fenthion,chlorpyrifos,and methyl parathion,with detection limits of 4×10-11 mol/L,2×10-9 mol/L,and 3×10-9 mol/L,respectively.In actual spinach sample testing,the spiked recoveries of the three OPs ranged from 86.5%to 110.6%.This study provided a new strategy for detecting pesticide residues using SERS.
7.Relationship between serum ROCK2,SDC-1,Apelin-13 expression and renal function and prognosis in patients with diabetes nephropathy
Hongjuan LI ; Wen LI ; Shuang REN ; Yinping MENG ; Liu ZHANG
International Journal of Laboratory Medicine 2025;46(22):2732-2737,2742
Objective To investigate the relationship between serum Rho-associated coiled coil containing protein kinase 2(ROCK2),syndecan-1(SDC-1),Apelin-13 and renal function and prognosis in patients with diabetes nephropathy.Methods From November 2019 to November 2021,153 patients with diabetic nephrop-athy admitted to the hospital were selected as the study group,and 153 patients with simple diabetes were se-lected as the simple diabetes group.Another 153 healthy individuals who underwent physical examinations during the same period were selected as the healthy control group.The levels of serum ROCK2,SDC-1 and Apelin-13 were determined by enzyme-linked immunosorbent assay.Pearson correlation analysis was used to analyze the correlations between the levels of serum ROCK2,SDC-1,and Apelin-13 and the clinical data of pa-tients with urinary nephropathy.Univariate and multivariate Logistic regression analyses were conducted to analyze the influencing factors of poor prognosis in patients with diabetic nephropathy.The receiver operating characteristic(ROC)curve was drawn to analyze the predictive value of serum ROCK2,SDC-1 and Apelin-13 for poor prognosis of patients with diabetes nephropathy.Results Compared with the healthy control group,the levels of serum ROCK2 and SDC-1 in the simple diabetes group and the study group increased,the level of Apelin-13 decreased,the estimated glomerular filtration rate(eGFR)decreased,and the ratio of urinary albu-min to creatinine(UACR)increased,and the differences were statistically significant(P<0.05).The levels of serum ROCK2,SDC-1,Apelin-13 and UACR in the study group were higher than those in the simple diabe-tes group,while eGFR was lower than that in the simple diabetes group,and the differences were statistically significant(P<0.05).Pearson correlation analysis indicated that the levels of serum ROCK2,SDC-1,and Apelin-13 in the study group were negatively correlated with eGFR(P<0.05),and positively correlated with renal disease stage,creatinine,24-hour proteinuria,and UACR index(P<0.05).Compared with the good prognosis group,the levels of serum ROCK2,DCC-1,Apelin-13 and UACR in the poor prognosis group in-creased(P<0.05),and eGFR decreased(P<0.05).Univariate and multivariate Logistic regression analyses indicated that the levels of serum ROCK2,SDC-1,and Apelin-13 were all influencing factors for the prognosis of patients(P<0.05).ROC curve analysis indicated that the area under the curve(AUC)for the combined prediction of poor prognosis in patients by serum ROCK2,DCC-1,and Apelin-13 levels was significantly grea-ter than that by ROCK2(Z=2.854,P=0.004)and DCC-1(Z=2.426,P=0.015)and Apelin-13(Z=2.172,P=0.030)were predicted separately.Conclusion The serum ROCK2,DCC-1 and Apelin-13 levels in patients with diabetic nephropathy increase and are related to renal function.The combination of the three has a relatively high predictive efficacy for the prognosis of patients.
8.Study on the effects and metabonomics of compound Kuijie Ankang Decoction for ulcerative colitis in mice
Cong REN ; Lili TANG ; Ruijuan LUO ; Fanyan MENG ; Yuping SHU ; Hongwu TAO ; Shuang LI ; Yizhen LI ; Chengcheng YUAN ; Yuedong LIU
International Journal of Traditional Chinese Medicine 2025;47(9):1255-1263
Objective:To explore the therapeutic effect of compound Kuijie Ankang Decoction on ulcerative colitis (UC) model mice by non targeted metabonomics; To explore its mechanism.Compound Kuijie Ankang.Methods:The mice were randomly divided into blank control group, model group, Kuijie Ankang Decoction group and sulfasalazine group, with 12 mice in each group. Except the blank control group, the other groups were given 1.5% DSS solution for free drinking to prepare UC model. After successful modeling, Kuijie Ankang Decoction group was intragastrically administered with compound Kuijie Ankang Decoction of 9.68 g/kg, sulfasalazine group was intragastrically administered with sulfasalazine capsule suspension of 320 mg/kg, model group and blank control group were intragastrically administered with equal volume of purified water, once a day, for 7 consecutive days. The body mass and disease activity index (DAI) score of mice were measured. ELISA was used to measure the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in the colon tissue of mice; the protein expressions of Claudin-1 and Zo-1 in colon tissue were detected by immunofluorescence method. HE staining was used to observe the pathological changes in the colon, and UHPLC-OE-MS technology was used to analyze the endogenous metabolite structure of mouse colon tissue, differential metabolites and related metabolic pathways were screened.Results:Compared with the model group, the colon length in Kuijie Ankang Decoction group and sulfasalazine group increased ( P<0.01), the DAI score decreased ( P<0.01), the levels of TNF-α, IL-1β and IL-6 in colon tissue decreased ( P<0.01), the level of IL-10 increased ( P<0.01), and the average optical density of Claudin-1 and Zo-1 protein increased ( P<0.01 or P<0.05). Metabolomics analysis identified 26 potential differential metabolites, including nicotinamide adenine dinucleotide, guanine, gamma aminobutyric acid, and thiamine, affecting 26 key metabolic pathways, including lysine biosynthesis, thiamine metabolism, cysteine and methionine metabolism. Conclusion:Kuaijie Ankang Decoction may improve metabolites such as Gamma aminobutyric acid and thiamine through metabolic pathways such as lysine biosynthesis to alleviate inflammatory reactions, thereby exerting therapeutic effects on ulcerative colitis in mice.
9.Exon Sequencing of HNF1β in Chinese Patients with Early-Onset Diabetes
Siqian GONG ; Hong LIAN ; Yating LI ; Xiaoling CAI ; Wei LIU ; Yingying LUO ; Meng LI ; Si-min ZHANG ; Rui ZHANG ; Lingli ZHOU ; Yu ZHU ; Qian REN ; Xiuying ZHANG ; Jing CHEN ; Jing WU ; Xianghai ZHOU ; Xirui WANG ; Xueyao HAN ; Linong JI
Diabetes & Metabolism Journal 2025;49(2):321-330
Background:
Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients.
Methods:
Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines.
Results:
Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher.
Conclusion
MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.
10.Acute Inflammatory Pain Induces Sex-different Brain Alpha Activity in Anesthetized Rats Through Optically Pumped Magnetometer Magnetoencephalography
Meng-Meng MIAO ; Yu-Xuan REN ; Wen-Wei WU ; Yu ZHANG ; Chen PAN ; Xiang-Hong LIN ; Hui-Dan LIN ; Xiao-Wei CHEN
Progress in Biochemistry and Biophysics 2025;52(1):244-257
ObjectiveMagnetoencephalography (MEG), a non-invasive neuroimaging technique, meticulously captures the magnetic fields emanating from brain electrical activity. Compared with MEG based on superconducting quantum interference devices (SQUID), MEG based on optically pump magnetometer (OPM) has the advantages of higher sensitivity, better spatial resolution and lower cost. However, most of the current studies are clinical studies, and there is a lack of animal studies on MEG based on OPM technology. Pain, a multifaceted sensory and emotional phenomenon, induces intricate alterations in brain activity, exhibiting notable sex differences. Despite clinical revelations of pain-related neuronal activity through MEG, specific properties remain elusive, and comprehensive laboratory studies on pain-associated brain activity alterations are lacking. The aim of this study was to investigate the effects of inflammatory pain (induced by Complete Freund’s Adjuvant (CFA)) on brain activity in a rat model using the MEG technique, to analysis changes in brain activity during pain perception, and to explore sex differences in pain-related MEG signaling. MethodsThis study utilized adult male and female Sprague-Dawley rats. Inflammatory pain was induced via intraplantar injection of CFA (100 μl, 50% in saline) in the left hind paw, with control groups receiving saline. Pain behavior was assessed using von Frey filaments at baseline and 1 h post-injection. For MEG recording, anesthetized rats had an OPM positioned on their head within a magnetic shield, undergoing two 15-minute sessions: a 5-minute baseline followed by a 10-minute mechanical stimulation phase. Data analysis included artifact removal and time-frequency analysis of spontaneous brain activity using accumulated spectrograms, generating spectrograms focused on the 4-30 Hz frequency range. ResultsMEG recordings in anesthetized rats during resting states and hind paw mechanical stimulation were compared, before and after saline/CFA injections. Mechanical stimulation elevated alpha activity in both male and female rats pre- and post-saline/CFA injections. Saline/CFA injections augmented average power in both sexes compared to pre-injection states. Remarkably, female rats exhibited higher average spectral power 1 h after CFA injection than after saline injection during resting states. Furthermore, despite comparable pain thresholds measured by classical pain behavioral tests post-CFA treatment, female rats displayed higher average power than males in the resting state after CFA injection. ConclusionThese results imply an enhanced perception of inflammatory pain in female rats compared to their male counterparts. Our study exhibits sex differences in alpha activities following CFA injection, highlighting heightened brain alpha activity in female rats during acute inflammatory pain in the resting state. Our study provides a method for OPM-based MEG recordings to be used to study brain activity in anaesthetized animals. In addition, the findings of this study contribute to a deeper understanding of pain-related neural activity and pain sex differences.

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