1.New advances in the treatment of neonatal diabetes mellitus with sulfonylureas
Xiaoyan HU ; Jinbo XIANG ; Xiaoxia ZHU ; Zheng LI ; Tingting CAO ; Ting DING ; Ziran XU ; Jingbo LI ; Youjun YANG
China Pharmacy 2026;37(9):1236-1240
Neonatal diabetes mellitus (NDM) is a rare monogenic disorder primarily caused by insufficient insulin secretion resulting from mutations in the KCNJ11 and ABCC8 genes. Sulfonylureas, represented by glibenclamide, have become the standard therapy for this type of NDM by precisely closing the mutated ATP-sensitive potassium channels in pancreatic β cells, thereby restoring insulin secretion. Clinical studies confirm that sulfonylureas enable over 90% of patients to successfully transition from insulin to oral treatment, achieving long-term stable glycemic control and improving neurological outcomes to a certain extent. In terms of safety, severe hypoglycemia induced by sulfonylureas is relatively rare and gastrointestinal reactions are mild; moreover, sulfonylureas show good long-term tolerability, and have no adverse effects on child growth and development. In the future, by further refining the full-chain management pathway of “rapid genetic diagnosis-early intervention-specialized dosage forms-long-term follow-up”, the clinical application of sulfonylureas is expected to provide NDM patients with an optimized treatment regimen and maximize their health benefits.
2.Expression and significance of tumor necrosis factor alpha,nuclear factor kappaB and ionized calcium binding adaptor molecule-1 in the hippocampus of mice with aortic dissection
Hong MA ; Xueling DING ; Qi WANG ; Hui LYU ; Albusm ASYA ; Xinyi CHENG ; Xiang MA
Chinese Journal of Tissue Engineering Research 2026;30(4):858-863
BACKGROUND:Hippocampal injury caused by aortic coarctation has been poorly studied,and combined detection of tumor necrosis factor α,nuclear factor κB and ionized calcium binding adaptor molecule-1 expression in aortic dissection has not been reported.OBJECTIVE:To observe histomorphologic changes in the hippocampus of a mouse model of aortic dissection and investigate the expression and significance of tumor necrosis factor alpha,nuclear factor kappaB and ionized calcium binding adaptor molecule-1 in the hippocampus of aortic dissection mice.METHODS:Sixteen healthy 3-week-old male C57BL/6 mice were randomly divided into two groups:control group and aortic dissection group,with eight mice in each group.In the aortic dissection group,mice were given β-aminopropionitrile monofumarate as drinking water for 4 weeks,and the angiotensin Ⅱ microinfiltration pump was then implanted to establish an animal model of aortic dissection.Mice in the control group were given normal diet and water.After the model was established,the maximum diameter of the ascending aorta was measured,hematoxylin-eosin staining and EVG staining were performed to evaluate the model formation rate,and the levels of inflammatory factors tumor necrosis factor α and interleukin 6 in serum were detected by enzyme-linked immunosorbent assay.The hippocampus was dissected and stained with hematoxylin-eosin to observe the pathological changes of the hippocampus in brain sections.The protein expression of tumor necrosis factor α,nuclear factor κB and ionized calcium binding adaptor molecule-1 was detected by western blot analysis.RESULTS AND CONCLUSION:(1)Compared with the control group,the maximum diameter of the ascending aorta in the aortic dissection group was significantly enlarged.(2)Hematoxylin-eosin staining of the aorta showed obvious thickening of the middle aorta and destruction and disorder of the aortic wall structure in mice.Neurons in the CA1 and CA3 regions of mice were sparsely arranged,reduced in size,and showed pyknosis with deeply stained nuclei.(3)Serum levels of inflammatory factors tumor necrosis factor α and interleukin 6 were increased in the aortic dissection group compared with the control group(P<0.01).(4)The expression levels of tumor necrosis factor α,nuclear factor κB,phosphorylated nuclear factor κB,and ionized calcium binding adaptor molecule-1 in the hippocampus were increased in the aortic dissection group compared with the control group(P<0.05).To conclude,microglial activation and increased expression of tumor necrosis factor α and nuclear factor κB may be involved in hippocampal neuron injury in aortic dissection mice.
3.Expression and significance of tumor necrosis factor alpha,nuclear factor kappaB and ionized calcium binding adaptor molecule-1 in the hippocampus of mice with aortic dissection
Hong MA ; Xueling DING ; Qi WANG ; Hui LYU ; Albusm ASYA ; Xinyi CHENG ; Xiang MA
Chinese Journal of Tissue Engineering Research 2026;30(4):858-863
BACKGROUND:Hippocampal injury caused by aortic coarctation has been poorly studied,and combined detection of tumor necrosis factor α,nuclear factor κB and ionized calcium binding adaptor molecule-1 expression in aortic dissection has not been reported.OBJECTIVE:To observe histomorphologic changes in the hippocampus of a mouse model of aortic dissection and investigate the expression and significance of tumor necrosis factor alpha,nuclear factor kappaB and ionized calcium binding adaptor molecule-1 in the hippocampus of aortic dissection mice.METHODS:Sixteen healthy 3-week-old male C57BL/6 mice were randomly divided into two groups:control group and aortic dissection group,with eight mice in each group.In the aortic dissection group,mice were given β-aminopropionitrile monofumarate as drinking water for 4 weeks,and the angiotensin Ⅱ microinfiltration pump was then implanted to establish an animal model of aortic dissection.Mice in the control group were given normal diet and water.After the model was established,the maximum diameter of the ascending aorta was measured,hematoxylin-eosin staining and EVG staining were performed to evaluate the model formation rate,and the levels of inflammatory factors tumor necrosis factor α and interleukin 6 in serum were detected by enzyme-linked immunosorbent assay.The hippocampus was dissected and stained with hematoxylin-eosin to observe the pathological changes of the hippocampus in brain sections.The protein expression of tumor necrosis factor α,nuclear factor κB and ionized calcium binding adaptor molecule-1 was detected by western blot analysis.RESULTS AND CONCLUSION:(1)Compared with the control group,the maximum diameter of the ascending aorta in the aortic dissection group was significantly enlarged.(2)Hematoxylin-eosin staining of the aorta showed obvious thickening of the middle aorta and destruction and disorder of the aortic wall structure in mice.Neurons in the CA1 and CA3 regions of mice were sparsely arranged,reduced in size,and showed pyknosis with deeply stained nuclei.(3)Serum levels of inflammatory factors tumor necrosis factor α and interleukin 6 were increased in the aortic dissection group compared with the control group(P<0.01).(4)The expression levels of tumor necrosis factor α,nuclear factor κB,phosphorylated nuclear factor κB,and ionized calcium binding adaptor molecule-1 in the hippocampus were increased in the aortic dissection group compared with the control group(P<0.05).To conclude,microglial activation and increased expression of tumor necrosis factor α and nuclear factor κB may be involved in hippocampal neuron injury in aortic dissection mice.
4.Effects of SPBC1604.04 Gene Deletion on Mitotic Cell Dynamics in Schizosaccharomyces pombe
Jia-Ni XU ; Jia-Yi HE ; Lang-Lin ZHENG ; Shu-Rong HE ; Shuai MA ; Xiang DING ; Yi-Ling HOU
Progress in Biochemistry and Biophysics 2026;53(5):1471-1484
ObjectiveMitochondria are not only the central organelles responsible for cellular energy metabolism but also play essential roles in regulating cell cycle progression and cytoskeletal dynamics. In recent years, accumulating evidence has demonstrated that mitochondrial homeostasis is closely associated with mitotic progression and cytokinesis. Schizosaccharomyces pombe serves as a classical and well-established model organism. Because its cell cycle regulatory mechanisms are highly conserved throughout evolution, its genetic background is clearly defined, and experimental manipulation is efficient and convenient, it has been extensively applied in studies of cell growth, division, and reproductive mechanisms. The SPBC1604.04 gene encodes a previously uncharacterized mitochondrial carrier protein in Schizosaccharomyces pombe. This gene is located on chromosome II and spans 1 018 base pairs in length. It encodes a protein consisting of 238 amino acids with a predicted molecular mass of approximately 31.03 ku. Bioinformatic analysis predicts that this protein is responsible for the transport of thiamine pyrophosphate (TPP) into mitochondria. However, the effects of SPBC1604.04 gene deletion on mitotic cell dynamics under different temperature conditions have not been fully elucidated. MethodsThe SPBC1604.04 deletion strain of Schizosaccharomyces pombe was used as the experimental model. Fluorescent protein markers were constructed in the deletion background to label mitochondria, microtubules, actin, myosin, the nuclear envelope, and chromosomes. Live-cell imaging was performed using a TCS-SP8 laser scanning confocal microscope under normal temperature conditions (25℃) and heat stress conditions (37℃). Time-lapse microscopy was applied to dynamically monitor mitochondrial morphology and distribution, spindle assembly and elongation, chromosome segregation, as well as the formation and constriction of the actomyosin ring during cytokinesis. ImageJ software was used for quantitative measurements, including microtubule length during mitosis, spindle length at different mitotic stages, mitochondrial fluorescence intensity as an indicator of mitochondrial content, actomyosin ring length, nuclear envelope area, and chromosome segregation timing. Statistical analyses were conducted to compare phenotypic differences between the wild-type and SPBC1604.04 deletion strains at both temperature conditions. Through these analyses, we systematically investigated the impact of SPBC1604.04 deletion on mitotic cell dynamics in fission yeast under both normal physiological conditions and temperature stress. ResultsAt 25℃, compared with wild-type cells, the SPBC1604.04Δ strain exhibited a pronounced tendency toward mitochondrial fragmentation, accompanied by abnormal mitochondrial content and a significant reduction in mitochondrial fluorescence intensity. These observations suggest impaired mitochondrial homeostasis under normal growth conditions. In addition, the constriction time of actomyosin ring during cytokinesis was markedly prolonged, indicating that deletion of SPBC1604.04 affects the dynamics of the contractile machinery. However, no obvious defects were observed in spindle assembly, spindle elongation, or chromosome segregation. Under heat stress at 37℃, mitochondrial morphology in the SPBC1604.04Δ strain showed a tendency to recover toward a continuous tubular network structure. Mitochondrial content was restored, fluorescence intensity increased, and the constriction time of the actomyosin ring returned to levels comparable to those of wild-type cells. These results indicate that the mitotic defects observed at normal temperature are partially or fully alleviated under heat stress conditions. ConclusionThis study demonstrates that deletion of the SPBC1604.04 gene leads to abnormal mitochondrial content in Schizosaccharomyces pombe. The mitochondrial carrier protein SPBC1604.04 participates in regulating actomyosin ring constriction during mitosis but does not appear to be directly involved in the regulation of spindle dynamics or chromosome segregation. Our findings provide key experimental evidence for understanding the functional link between the SPBC1604.04 gene, mitochondrial homeostasis, and mitotic regulation.
5.Effects of SPBC1604.04 Gene Deletion on Mitotic Cell Dynamics in Schizosaccharomyces pombe
Jia-Ni XU ; Jia-Yi HE ; Lang-Lin ZHENG ; Shu-Rong HE ; Shuai MA ; Xiang DING ; Yi-Ling HOU
Progress in Biochemistry and Biophysics 2026;53(5):1471-1484
ObjectiveMitochondria are not only the central organelles responsible for cellular energy metabolism but also play essential roles in regulating cell cycle progression and cytoskeletal dynamics. In recent years, accumulating evidence has demonstrated that mitochondrial homeostasis is closely associated with mitotic progression and cytokinesis. Schizosaccharomyces pombe serves as a classical and well-established model organism. Because its cell cycle regulatory mechanisms are highly conserved throughout evolution, its genetic background is clearly defined, and experimental manipulation is efficient and convenient, it has been extensively applied in studies of cell growth, division, and reproductive mechanisms. The SPBC1604.04 gene encodes a previously uncharacterized mitochondrial carrier protein in Schizosaccharomyces pombe. This gene is located on chromosome II and spans 1 018 base pairs in length. It encodes a protein consisting of 238 amino acids with a predicted molecular mass of approximately 31.03 ku. Bioinformatic analysis predicts that this protein is responsible for the transport of thiamine pyrophosphate (TPP) into mitochondria. However, the effects of SPBC1604.04 gene deletion on mitotic cell dynamics under different temperature conditions have not been fully elucidated. MethodsThe SPBC1604.04 deletion strain of Schizosaccharomyces pombe was used as the experimental model. Fluorescent protein markers were constructed in the deletion background to label mitochondria, microtubules, actin, myosin, the nuclear envelope, and chromosomes. Live-cell imaging was performed using a TCS-SP8 laser scanning confocal microscope under normal temperature conditions (25℃) and heat stress conditions (37℃). Time-lapse microscopy was applied to dynamically monitor mitochondrial morphology and distribution, spindle assembly and elongation, chromosome segregation, as well as the formation and constriction of the actomyosin ring during cytokinesis. ImageJ software was used for quantitative measurements, including microtubule length during mitosis, spindle length at different mitotic stages, mitochondrial fluorescence intensity as an indicator of mitochondrial content, actomyosin ring length, nuclear envelope area, and chromosome segregation timing. Statistical analyses were conducted to compare phenotypic differences between the wild-type and SPBC1604.04 deletion strains at both temperature conditions. Through these analyses, we systematically investigated the impact of SPBC1604.04 deletion on mitotic cell dynamics in fission yeast under both normal physiological conditions and temperature stress. ResultsAt 25℃, compared with wild-type cells, the SPBC1604.04Δ strain exhibited a pronounced tendency toward mitochondrial fragmentation, accompanied by abnormal mitochondrial content and a significant reduction in mitochondrial fluorescence intensity. These observations suggest impaired mitochondrial homeostasis under normal growth conditions. In addition, the constriction time of actomyosin ring during cytokinesis was markedly prolonged, indicating that deletion of SPBC1604.04 affects the dynamics of the contractile machinery. However, no obvious defects were observed in spindle assembly, spindle elongation, or chromosome segregation. Under heat stress at 37℃, mitochondrial morphology in the SPBC1604.04Δ strain showed a tendency to recover toward a continuous tubular network structure. Mitochondrial content was restored, fluorescence intensity increased, and the constriction time of the actomyosin ring returned to levels comparable to those of wild-type cells. These results indicate that the mitotic defects observed at normal temperature are partially or fully alleviated under heat stress conditions. ConclusionThis study demonstrates that deletion of the SPBC1604.04 gene leads to abnormal mitochondrial content in Schizosaccharomyces pombe. The mitochondrial carrier protein SPBC1604.04 participates in regulating actomyosin ring constriction during mitosis but does not appear to be directly involved in the regulation of spindle dynamics or chromosome segregation. Our findings provide key experimental evidence for understanding the functional link between the SPBC1604.04 gene, mitochondrial homeostasis, and mitotic regulation.
6.Umbrella decision-making model for diagnosis and treatment of elderly lung cancer patients: Construction and practice
Lunxu LIU ; Jian ZHOU ; Xiang DING ; Nan CHEN ; Jianxin XUE ; Xuelei MA ; Ye WANG ; Weiya WANG ; Liqing PENG ; Xin YOU ; Minggang SU ; Xu CHENG ; Jiao WANG ; Ning GE ; Deying KANG ; Yuchen HUANG ; Jinghan WANG ; Yu TONG ; Yaoxi ZHANG ; Jirong YUE ; Hu LIAO
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(06):833-839
With the accelerating trend of population aging, the number of elderly patients with lung cancer continues to rise, and the disease burden is becoming increasingly heavy. The clinical management of these patients faces severe challenges due to their decreased physiological reserve, complex comorbidities, and significant individual heterogeneity. Consequently, under traditional diagnosis and treatment models, doctors often struggle to identify the individualized risks of elderly patients in a timely and comprehensive manner, which can easily lead to decision biases such as undertreatment or overtreatment. In view of this, this study advocates for the establishment of an umbrella decision-making model specifically tailored for elderly lung cancer patients. Grounded in a multidisciplinary team (MDT) platform, this model deeply integrates oncological indicators with the comprehensive geriatric assessment (CGA) system. By holistically considering multidimensional variables including tumor burden, organ function, frailty index, cognitive status, and social support, the model establishes an operational mechanism characterized by "single entry, precise stratification, and targeted selection". Accordingly, patients can be scientifically triaged into distinct intervention tiers, such as active surveillance, minimally invasive surgery, drug therapy, radiotherapy, and best supportive care, thereby achieving real-time alignment between treatment intensity and patient fitness. This article elaborates on the construction logic and key operational procedures of this novel decision-making framework, aiming to guide clinical practice beyond the limitations of a tumor-centric perspective toward a holistic, dynamic, whole-course management strategy. This transition seeks to ensure optimal quality of life and clinical net benefit for elderly patients alongside survival prolongation.
7.Cationic nanoemulsions on the tear film of orthokeratology wearers
Xiang SI ; Peiran ZHANG ; Lei DING ; Haixia HU ; Lei YIN ; Shuqin LI
International Eye Science 2026;26(8):1343-1347
AIM:To observe the effect of cationic nanoemulsion eye drops on the tear film and ocular surface morphology of orthokeratology wearers, and to provide a basis for the clinical selection of eye drops during orthokeratology wear. METHODS:A prospective study was conducted on patients with refractive errors who were fitted with orthokeratology lenses for the first time at Anhui Aier Eye Hospital,from February 2025 to October 2025. Patients were randomly assigned to the experimental group, which received a cationic nanoemulsion eye drops, or the control group, which received a 0.1% sodium hyaluronate eye drops. One drop of the assigned lubricant was instilled in each eye before lens insertion and before lens removal in both groups for 3 consecutive months. Uncorrected visual acuity, spherical equivalent, anterior corneal curvature, tear film break-up time, tear film lipid layer score, and corneal fluorescein staining score were compared between the two groups at baseline and at 1 wk, 1 and 3 mo after lens wear. RESULTS: A total of 64 eyes(all right eyes)from 64 patients with refractive errors were enrolled. Ultimately, 60 patients completed the final follow-up, including 30 in the experimental group(n=30; male/female: 14/16; age: 9.57±1.07 y)and the control group(n=30; male/female: 12/18; age: 9.43±1.01 y). No significant intergroup differences were found in uncorrected visual acuity, spherical equivalent, anterior corneal curvature, or corneal fluorescein staining scores either before or after lens wear(all P>0.05).The tear film break-up time was significantly longer and tear film lipid layer score was significantly better in the experimental group than in the control group at post-wear time point(all P<0.05). Uncorrected visual acuity, spherical equivalent, and anterior corneal curvature improved significantly from baseline at 1 wk, 1 and 3 mo after lens wear in both groups(all P<0.05). Corneal fluorescein staining scores increased significantly from baseline at 1 wk and 1 mo in the control group(both P<0.05), whereas no significant change was observed in the experimental group. No cases of severe corneal injury(score of 3)were found in either group. CONCLUSION: Cationic nanoemulsion eye drops have a similar effect to 0.1% sodium hyaluronate eye drops in improving the visual acuity and corneal morphology of orthokeratology wearers, but have more advantages in stabilizing the tear film and improving the state of the tear film lipid layer, and have good safety, so they can be used as an ideal choice of eye drops for orthokeratology wearers.
8.Application of contrast-enhanced ultrasound in the full-course management of precise locoregional diagnosis and treatment for hepatocellular carcinoma
Yan ZHOU ; Jianmin DING ; Xiang JING
Journal of Clinical Hepatology 2026;42(7):1513-1518
Hepatocellular carcinoma (HCC) is one of the major malignancies with the heaviest disease burden in China. Many patients have lost the opportunity for radical surgical resection at the time of diagnosis, with locoregional interventional therapy playing an increasingly important role in full-course HCC management. Contrast-enhanced ultrasound (CEUS) enables real-time dynamic assessment of tumor microvascular perfusion and has several practical advantages including bedside availability, no ionizing radiation, and repeatability, and its role has therefore expanded from an adjunctive diagnostic method to the full-course management of precise locoregional diagnosis and treatment for HCC. Before treatment, CEUS can assist in the qualitative diagnosis of lesions, lesion localization, delineation of the viable tumor, and planning of the puncture route, especially for lesions that are poorly visualized on conventional ultrasound. During and after treatment, CEUS can help to detect residual viable tumor, related complications, and local tumor progression, thereby providing a basis for additional ablation, transcatheter arterial chemoembolization, or adjustment of combined treatment strategies. In recent years, the CEUS LI-RADS treatment response assessment system has provided a standardized framework for evaluating response after locoregional therapy, and in addition, the development of unimodal and multimodal fusion imaging techniques has further promoted the standardized application of CEUS in precise locoregional therapy for HCC. As increasing attention is paid to the ablation margin, the goal of response evaluation for local ablation has shifted from “the achievement of imaging-defined complete ablation” to a new treatment paradigm of “no contrast-enhanced imaging, no ablation; no fusion imaging, no curative ablation”. This article reviews the technical advantages, clinical application value, treatment response assessment, fusion imaging strategies, and current limitations of CEUS in locoregional interventional diagnosis and treatment of HCC, in order to provide a reference for the individualized and precise management of HCC.
9.Preparation and performance evaluation of dissolving microneedles with a single matrix of Pterocarya hupehensis total flavonoids
Xingxing GAO ; Dan CHEN ; Yan XIA ; Lin DING ; Xingyue CHEN ; Yang XIANG ; Lunzhi LIU ; Lin YUAN
China Pharmacy 2026;37(16):2097-2104
OBJECTIVE To screen a single matrix, prepare dissolving microneedles of Pterocarya hupehensis total flavonoids(PHTF-MN), and characterize it while evaluating its analgesic effects.METHODS Using microneedle morphology and puncture rate as evaluation indicators, the matrix was screened from polyvinyl alcohol (PVA), polyvinylpyrrolidone K30 (PVP K30), PVP K90, and sodium hyaluronate with relative molecular weights of 15 000 and 60 000 (HA 1.5w and HA 6w ). PHTF-MN was prepared by the layer-by-layer casting method with the selected matrix, and the appearance and morphology, total flavonoid content, mechanical properties, puncture performance, dissolution properties, and drug release behavior of the obtained PHTF-MN were characterized. Using male BALB/c mice as test subjects and aspirin (ASA) as a positive control, the analgesic effects of the microneedles were evaluated using a formalin-induced pain model.RESULTS Blank microneedles prepared with 30% HA 1.5w exhibited optimal surface flatness and the membrane puncture rate (through 4 layers of parafilm membrane) reaching 78%. The PHTF-MNs (with PHTF contents of 10, 20, 30, 40, and 50 mg/g in the matrix) exhibited uniform arrays and intact tips, with RSDs for total flavonoid contents all below 7% ( n =8). The single-needle puncture forces were 1.46 and 1.07 N (with PHTF contents of 30 and 50 mg/g in the matrix, respectively). The skin penetration rate approached 100%, with visible puncture marks in the stratum corneum and drug residue (with a PHTF content of 50 mg/g in the matrix). After 30 min of fixation, the needles had almost completely dissolved, and the skin retention rates all exceeded 90% (with PHTF contents of 10, 30, and 50 mg/g in the matrix, respectively). The cumulative release within 8 h was (134.64±2.03) μg/cm 2 , and the fit to the first-order kinetic equation ( R 2 =0.950 1) was highest (with a PHTF content of 50 mg/g in the matrix). Low- and high-dose PHTF-MN (containing 85.26 and 145.03 μg of total flavonoids per patch, respectively; 1 patch/day) significantly shortened the cumulative time of phase Ⅱ pain response in mice ( P <0.05), with pain inhibition rates (43.67% and 59.78%) comparable to those of ASA (53.70%).CONCLUSIONS PHTF-MN using HA 1.5w as a single matrix was successfully prepared. This formulation features a simplified preparation process, uniform drug loading, favorable mechanical properties, resistance to fracture, and rapid dissolution. Its drug release behavior conforms to a first-order kinetic model, and its analgesic effect is comparable to that of ASA.
10.Effect of Huangqin Qingre Chubi Capsules-containing Serum on CircRNA_0001543/NF-κB Expression in Co-cultured PBMCs and Human FLSs from Patients with Ankylosing Spondylitis
Yajun QI ; Jian LIU ; Qiao ZHOU ; Yuedi HU ; Xiang DING ; Chengzhi CONG ; Xu LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):87-95
ObjectiveThis study aims to explore the effects of Huangqin Qingre Chubi capsules-containing serum on the expression of CircRNA_0001543/nuclear factor-kappa B (NF-κB) in co-cultured peripheral blood mononuclear cells (PBMCs) and human fibroblast-like synoviocytes (FLSs) from patients with ankylosing spondylitis (AS). MethodsVenous blood was collected from patients with AS to isolate PBMCs. FLSs were co-cultured with AS patients' PBMCs, and FLSs were harvested after co-culture for subsequent experiments. The normal control group consisted of normal FLSs, while the model group comprised co-cultured AS PBMCs and FLSs to simulate AS pathology. The Huangqin Qingre Chubi capsules group involved adding Huangqin Qingre Chubi capsules-containing serum to the co-cultured cells(6.48 g·kg-1). To investigate the effect of HQC-containing serum on the viability of co-cultured cells, and the experiment was divided into the following groups based on the dilution concentration: blank group, 10% HQC group, 20% HQC group, and 30% HQC group.To study the influence of the optimal concentration of HQC-containing serum on cytokine and pathway indicators in each group, the experiment was divided into three groups: normal group, model group, and optimal concentration HQC-containing serum group.For the validation of the transfection efficiency of the CircRNA_0001543 interference plasmid, the experiment was divided into the following groups: blank group, si-NC group (with transfection reagent), si-circ_0001543-1 group (with transfection reagent and interference plasmid No. 1 targeting circ_0001543), si-circ_0001543-2 group (with transfection reagent and interference plasmid No. 2 targeting circ_0001543), and si-circ_0001543-3 group (with transfection reagent and interference plasmid No. 3 targeting circ_0001543).For the validation of the transfection efficiency of the CircRNA_0001543 overexpression plasmid, the experiment was divided into the following groups: blank group, OE-NC group (with transfection reagent), and OE-circ_0001543 group (with transfection reagent and overexpression plasmid targeting circ_0001543).To study the effects of CircRNA_0001543 interference/overexpression on cytokine and pathway indicators in each group, the experiment was divided into the following groups: si-NC group, si-CircRNA_0001543 group, OE-NC group, and OE-CircRNA_0001543 group. Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of interleukin-1β (IL-1β), IL-10, IL-37, and tumor necrosis factor-α (TNF-α). Real-time quantitative polymerase chain reaction (Real-time PCR) was utilized to measure the expression of CircRNA_0001543, IκBα, and NF-κB p65. ResultsAfter 48 hours, 30% Huangqin Qingre Chubi Capsules-containing serum significantly inhibited the proliferation of co-cultured PBMCs and FLSs, which was determined to be the optimal experimental drug-containing serum concentration. Compared with those in the normal group, the expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α in the model group were significantly increased (P<0.01), while the expressions of CircRNA_0001543 mRNA, IL-10, and IL-37 were significantly decreased (P<0.01). Compared with those in the model group, the expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α in the Huangqin Qingre Chubi Capsules-containing serum group were significantly decreased (P<0.05), and the expressions of CircRNA_0001543 mRNA, IL-10, and IL-37 were significantly increased (P<0.05), with the most prominent changes in the 30% drug-containing serum group (P<0.01). Compared with that in the si-NC group, the expression of CircRNA_0001543 was significantly reduced in the si-CircRNA_0001543 group (P<0.01). Compared with that in the OE-NC group, the expression of CircRNA_0001543 was significantly increased in the OE-CircRNA_0001543 group (P<0.01), indicating that the si-CircRNA_0001543 and OE-CircRNA_0001543 plasmids were successfully transfected. Based on the optimal drug-containing serum of Huangqin Qingre Chubi Capsules, si-CircRNA_0001543 transfection led to significantly increased expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α and decreased the expressions of IL-10 and IL-37 (P<0.01). In contrast, OE-CircRNA_0001543 transfection significantly decreased the expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α (P<0.01) and increased the expressions of IL-10 and IL-37 (P<0.01). ConclusionHuangqin Qingre Chubi capsules-containing serum can improve immune inflammation in AS by increasing the expression of CircRNA_0001543, regulating the NF-κB pathway, suppressing pro-inflammatory cytokines, and enhancing anti-inflammatory cytokine expression.

Result Analysis
Print
Save
E-mail