1.Pathogenesis and Syndrome Differentiation of "Gaozhuo" of Oxidative Stress in Diabetic Kidney Disease
Yuman YIN ; Yunfeng YU ; Xiangning HUANG ; Jiawang HUANG ; Gang HU ; Juan HUANG ; Rong YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):226-234
Oxidative stress is a pivotal factor in the onset and progression of diabetic kidney disease (DKD), and it plays an essential role in the prevention and treatment of DKD. The "Gaozhuo" pathogenesis posits that DKD is characterized by the invasion of Gaozhuo and damage to the kidney collaterals, with the underlying cause being the insufficiency of spleen Qi and the internal formation of Gaozhuo, which provides valuable guidance on oxidative stress. The insufficiency of spleen Qi and the internal formation of Gaozhuo represent a dynamic, evolving process. Gaozhuo invades the kidney collaterals, impairs kidney Qi, and progressively leads to the congealing and stagnation of Gaozhuo and blood, ultimately resulting in the failure of both the spleen and kidneys. The damage caused by Gaozhuo to the kidney collaterals and kidney Qi is analogous to the organ and functional damage of the kidneys induced by excessive reactive oxygen species and oxidative stress. Damage to the kidney collaterals means organic injuries to the glomeruli, renal tubules, and renal interstitium, and the depletion of kidney Qi refers to damage to glomerular filtration and renal tubular reabsorption. The congealing and stagnation of Gaozhuo and blood in the kidney collaterals is similar to oxidative stress-induced thickening of the glomerular basement membrane and fibrosis. The interaction between spleen and kidney Qi deficiency and the congealing and stagnation of Gaozhuo and blood creates a vicious cycle that exacerbates the condition, ultimately evolving into the failure of both the spleen and kidneys. The failure of the spleen and kidneys is analogous to renal failure, and its extreme manifestation is end-stage renal disease and uremia. The treatment of oxidative stress in DKD with traditional Chinese medicine (TCM) is based on the principles of strengthening the spleen and tonifying the kidneys, and dispelling turbidity and removing blood stasis. According to the syndrome type, it is recommended to use methods such as strengthening the spleen and tonifying Qi while dispelling dampness and removing turbidity, strengthening the spleen and tonifying the kidneys while dispelling dampness and removing turbidity, strengthening the spleen and tonifying the kidneys while dispelling turbidity and removing blood stasis, or consolidating the spleen and kidneys while clearing away turbidity and blood stasis.
2.Shashen Maidong Tang Enhances Efficacy of Chemotherapy in Mouse Model of Lewis Lung Cancer by Modulating JAK2/STAT3 Signaling Pathway
Lin YU ; Yaoyao WANG ; Limin LIU ; Zuowei HU ; Yanping ZHOU ; Shang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):1-10
ObjectiveTo predict the mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer via network pharmacology and validate the prediction results in animal experiments. MethodsThe potential mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer was predicted by network pharmacology, liquid chromatography-mass spectrometry (LC-MS), and molecular docking methods. C57/BL6 mice were assigned into normal, model, cisplatin, and Shasheng Maidong Tang+cisplatin groups. In addition to the normal group, the remaining groups were injected subcutaneously with 0.2 mL of 1×107 cells·mL-1 Lewis lung cancer cells to establish the Lewis lung cancer model. The daily gavage dose of Shasheng Maidong Tang was 3.58 g·kg-1, and the concentration of cisplatin intraperitoneally injected on every other day was 2 mg·kg-1. Drugs were administered for 14 d. The changes in the tumor volume and the rate of tumor suppression were monitored, and the tumor histopathological changes were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay was employed to measure the interleukin (IL)-6 and interferon (IFN)-γ levels in peripheral blood. Real-time PCR was performed to quantify the mRNA levels of Janus kinase 2 (JAK2), signal transducer and activator of transcription 1 (STAT1), and signal transducer and activator of transcription 3 (STAT3) in the tumor tissue of mice. Western blot was employed to determine the protein levels of JAK2, STAT3, B-cell lymphoma-2 (Bcl-2), cysteinyl aspartate-specific proteinase-3 (Caspase-3), and Pim-1 proto1 (PIM1) in the tumor tissue. Immunohistochemistry was employed to detect the expression of Bcl-2 and PIM1 in the tumor tissue. ResultsNetwork pharmacological predictions indicated that Shasheng Maidong Tang might enhance the efficacy of chemotherapy for lung cancer by regulating nitrogen metabolism, AGE-RAGE signaling pathway, cancer pathway, and JAK/STAT signaling pathway. The experimental results demonstrated that tumor volume in the cisplatin group and Shasheng Maidong Tang+cisplatin group was reduced compared with the model group, with statistically distinct differences observed on days 14, 17, 20 post modeling (P<0.05). Notably, the Shasheng Maidong Tang+cisplatin therapy further decreased tumor volume compared with the cisplatin group, showing marked reductions on days 17 and 20 (P<0.05), consistent with trends visualized in tumor volume comparison charts. The Shasheng Maidong Tang+cisplatin group exhibited higher tumor inhibition rate than the cisplatin group (P<0.05). Histopathological analysis via HE staining revealed that the tumors in the model group displayed frequent nuclear mitosis, densely arranged cells, hyperchromatic nuclei, and no necrosis. Cisplatin treatment induced partial necrosis and vacuolization, while the Shasheng Maidong Tang+cisplatin group exhibited extensive necrotic regions, maximal vacuolization, disarranged tumor cells, and minimal mitotic activity. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed elevated level of IFN-γ (P<0.01) and declined level of IL-6 (P<0.01) in the peripheral blood. Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented elevated level of IFN-γ (P<0.01) and lowered level of IL-6 (P<0.01) in the peripheral blood. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin groups showed down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level STAT1 (P<0.01). Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level of STAT1 (P<0.01). Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), and STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). Compared with the cisplatin group, Shasheng Maidong Tang+cisplatin group presented down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). The Bcl-2 and PIM1 expression results obtained by immunohistochemistry were consistent with those of Western blot. ConclusionShasheng Maidong Tang may enhance the efficacy of chemotherapy in the mouse model of Lewis lung cancer by regulating the JAK2/STAT3 signaling pathway.
3.Effect of summer short-term clinical practice teaching on empathy ability in rehabilitation therapy students
Chinese Journal of Rehabilitation Theory and Practice 2026;32(1):117-124
ObjectiveTo explore an centralized clinical practice teaching model delivered via a summer short-term semester, and to examine its effectiveness on empathy and communication efficacy among undergraduate students majoring in rehabilitation therapy through a mixed-methods design. MethodsFrom June to July, 2025, forty second-year rehabilitation therapy undergraduates from Nanchang Medical College participated in a one-week immersive clinical practice during the summer short-term semester. An action research framework integrating a one-group pre-post experimental design and qualitative research methods were adopted. Quantitative data were collected using the Chinese-Adapted Jefferson Scale of Empathy-Medical Student Version (JSE-HP) to assess changes in empathy. Qualitative data were obtained through semi-structured focus group interviews and structured reflective journals to investigate students' experiences and transformations in empathic cognition, emotional integration and professional identity. ResultsAfter the teaching intervention, students demonstrated significant improvements in the total score of JSE-HP and all subdimensions (perspective taking, compassionate care and standing in the patient's shoes) (t < -3.69, P < 0.01). Qualitative analysis yielded three core themes: reconstruction of clinical reasoning paradigms, emotional-cognitive integration and elevation of professional identity. ConclusionSummer short-term semester clinical practice model, structured around “clinical immersion, narrative reflection, interprofessional collaboration and formative assessment”, effectively facilitates embodied cultivation of empathy. This model not only bridges the gap between theory and clinical practice, but also serves as an educational catalyst for students' transformation from technical performers to humanistic caregivers. It aligns with the core concepts of the World Health Organization rehabilitation competency framework, offering a replicable and scalable approach to advancing systematic reform in medical humanities education.
4.Differences in chemical components and quality analysis of Gardenia jasminoides before and after processing with ginger
Lihua TANG ; Yu WU ; Xuedi HUANG ; Xiaolian HU ; Yi TANG ; Zilong CHEN ; Xiaofan XIAO ; Xide YE
China Pharmacy 2026;37(2):168-173
OBJECTIVE To analyze the differences in chemical components of Gardenia jasminoides before and after processing with ginger, and to evaluate the quality differences among different producing areas. METHODS Ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry was used to analyze the compositional differences of G. jasminoides before and after processing with ginger. The water content, total ash, and ethanol-soluble extract content of ginger- processed G. jasminoides were determined according to the 2020 edition of Chinese Pharmacopoeia. High performance liquid chromatography was adopted to determine the contents of genipin gentiobioside, geniposide, crocin Ⅰ and crocin Ⅱ in ginger- processed G. jasminoides. RESULTS A total of 49 chemical components were identified from raw G. jasminoides and ginger- processed G. jasminoides, including 14 flavonoids, 15 iridoids, 10 organic acids, 2 alkaloids and 8 other compounds. Among them, 42 components were detected in raw G. jasminoides, 28 in ginger-processed G. jasminoides, and 21 components were common to both. After processing with ginger, raw G. jasminoides lost 21 components (including iridoids, flavonoids, alkaloids, and others), while 7 chemical components were added (including coumarins, organic acids, organic acid esters, and flavonoids). For the 15 batches of ginger-processed G. jasminoides, the water content ranged from 5.64% to 7.11%, total ash from 2.92% to 4.87%, and ethanol-soluble extract from 40.61% to 58.02%. The average contents of genipin gentiobioside, geniposide, crocin Ⅰ and crocin Ⅱ were 0.108 7, 0.542 2, 0.565 0, and 0.012 5 mg/g, respectively. CONCLUSIONS After processing with ginger, G. jasminoides loses 21 components, while 7 new components are added. Differences are observed in the water content, total ash, ethanol-soluble extract, and the contents of genipin gentiobioside, geniposide, crocin Ⅰ, and crocin Ⅱ of ginger-processed G. jasminoides from different producing areas. Notably, samples from Fujian exhibit high contents of genipin gentiobioside and ethanol-soluble extract, while samples from Jiangxi have a high content of crocin Ⅰ.
5.Efficacy and Application Characteristics of Cold Chinese Medicines Based on Chinese Pharmacopoeia (2020 Edition)
Lu YUE ; Yilong HU ; Jingying YANG ; Xiangxiang WU ; Mingsan MIAO ; Ming BAI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):241-248
ObjectiveTo provide a reference for the rational clinical use of cold Chinese medicines by sorting and analyzing their properties, flavors, meridian tropism, primary therapeutic indications, methods of administration, dosages, and precautions as recorded in the 2020 edition of Pharmacopoeia of the People's Republic of China (Chinese Pharmacopoeia). MethodsCold Chinese medicines for internal and external use included in the 2020 edition of Chinese Pharmacopoeia were entered one by one, and their efficacy, properties, flavors, meridian tropism, methods of administration, dosages, and usage precautions were statistically classified and summarized to guide clinical medication use. ResultsA total of 259 cold Chinese medicines for internal use were included and categorized into 18 efficacy groups, mainly comprising heat-clearing drugs, water-excreting and dampness-draining drugs, and phlegm-resolving, cough- and asthma-relieving drugs. Their predominant flavors were bitter, sweet, and pungent, and they primarily entered the liver, lung, and stomach meridians. The main methods of administration included decocting first, grinding into powder for oral use, or preparing into pills or powders, with most dosages ranging from 9 to 15 g. A total of 83 cold Chinese medicines for external use were included, involving 16 efficacy categories. Their main flavors were bitter, sweet, and pungent, primarily entering the liver, lung, and large intestine meridians. The main external application methods were grinding into powder for topical use or preparing decoctions for fumigation and washing, with most dosages ranging from 9 to 15 g. Whether for internal or external use, cold Chinese medicines should be used with caution or contraindicated in pregnant women. ConclusionThe cold Chinese medicines included in the 2020 edition of the Chinese Pharmacopoeia are mainly suitable for patients with carbuncles, swellings, and coughs. However, in clinical practice, it is necessary to strictly follow the principles of syndrome differentiation and treatment, pay attention to administration methods and dosages, and use cold medicines rationally and effectively to improve clinical efficacy.
6.Comparison of sleep EEG power spectral density between depressive episode patients and schizophrenia patients with suicidal behavior
Jingwen LIU ; Yunfei ZHOU ; Jingchu HU ; Jiaoyan ZHOU ; Junwei YANG ; Jie LIANG ; Hong XU ; Yu CANG ; Shimeng MA
Sichuan Mental Health 2026;39(1):50-57
BackgroundPatients with depressive episode and schizophrenia have a high risk of suicide. The sleep electroencephalogram power spectral density characteristics of patients with depressive episode accompanied by suicidal behavior and those with schizophrenia may be different, but there is currently a lack of direct comparative studies on these two groups of patients. ObjectiveTo compare the sleep electroencephalogram power spectral density between depressive episode and schizophrenic patients with suicidal behavior, in order to provide references for exploring predictive indicators of suicidal behavior. MethodsFrom June 2018 to December 2020, 20 patients with depressive episode and 20 patients with schizophrenia who had committed suicide within the past month and were treated at the outpatient department of Shenzhen Kangning Hospital were selected. All of them met the diagnostic criteria for depressive episode or schizophrenia as defined in the International Classification of Diseases, tenth edition (ICD-10). Using a random sampling method, 20 volunteers with matching gender and age to the patient groups were selected from the Cuiping community in Shenzhen as the control group. The subjective sleep of the patients was evaluated using the Insomnia Severity Index (ISI), the Dysfunctional Belief and Attitude about Sleep (DBAS), the Disturbing Dreams and Nightmare Severity Index (DDNSI), and the Epworth Somnolence Scale (ESS). The objective sleep of the patients was assessed using polysomnography. The sleep electroencephalogram was filtered and the power spectral density of the brain wave was analyzed and processed for all the subjects. The subjective and objective sleep conditions of the two patient groups were compared, and the sleep electroencephalogram power spectral density of the patient groups and the control group were also compared. ResultsA comparison of subjective and objective sleep conditions between patients with depressive episode accompanied by suicidal behavior and patients with schizophrenia accompanied by suicidal behavior showed no statistically significant differences (P>0.05). Comparisons of sleep electroencephalogram power spectral density in the W stage (average power of α wave, total power of δ wave, average power of δ wave, average power of θ wave), N1 stage (average power of β wave, total power of α wave, total power of δ wave), N2 stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave), N3 stage (average power of α wave, average power of δ wave), and R stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave) between patients with depressive episode accompanied by suicidal behavior, patients with schizophrenia accompanied by suicidal behavior, and the control group showed statistically significant differences (P<0.05 or 0.01). The total power of δ wave in the W stage and the average power of β wave and δ wave in the N1 stage were higher in two patient groups were higher than those of the control group. The total power of α wave and the average power of α wave in the N2 stage were lower than those of the control group, while the average power of δ wave was higher than that of the control group. The average power of α wave in the N3 stage of both patient groups were lower than that of the control group, while the average power of δ wave was higher than that of the control group. The total power and average power of α wave in the R stage were lower than those of the control group, while the total power and average power of δ wave were higher than those of the control group. All the differences were statistically significant. Patients with depressive episode accompanied by suicidal behavior had higher average powers of α wave, δ wave, and θ wave in the W stage compared with the control group, while the total power of α wave in the N1 stage was lower in the former group. All these differences were statistically significant (P<0.05). ConclusionThe depressive episode patients accompanied by suicidal behavior have highly overlapping sleep electroencephalogram abnormal patterns with those of schizophrenia patients, mainly manifested as a general decrease in α wave power (N2, N3, R stage) and a general increase in δ wave power (W, N1, N2, N3, R stage) as well as β wave power in N1 stage. At the same time, patients with depressive episode accompanied by suicidal behavior also show specific changes, including an increase in the average power of α and θ waves during the wakefulness period (W stage), and a decrease in the total power of α wave in N1 stage. [Funded by Guangdong Province High-level Clinical Key Specialty (with supporting funds from Shenzhen City) (number, SZGSP013); Shenzhen Key Medical Discipline (number, SZXK041); Shenzhen Clinical Medicine Research Center Project (number, 20210617155253001)]
7.Effect of ultrasound-guided needle-knife release of the ligamentum flavum on the expression of integrin alpha5 and beta1 in degenerative rabbit lumbar intervertebral discs
Can CHEN ; Yu ZHAO ; Binhan HU ; Mengfan DU ; Junning LIU ; Susheng NIU ; Yan ZHANG
Chinese Journal of Tissue Engineering Research 2025;29(2):331-338
BACKGROUND:Needle-knife release of the ligamentum flavum can effectively improve symptoms in patients with lumbar degeneration,and ultrasound guidance can increase the precision of needle-knife release;however,the specific effects of needle-knife release of the ligamentum flavum on the degenerated intervertebral discs and the possible mechanisms remain to be clarified. OBJECTIVE:To investigate the effect of ultrasound-guided needle-knife release of the ligamentum flavum. METHODS:Twenty-four New Zealand rabbits were randomized into control(n=6)and model(n=18)groups.A rabbit model of lumbar disc degeneration model was established in the model group by cutting the supraspinous and interspinous ligaments of the L5/6 and L6/7 segments to maintain a standing posture and apply axial load to the lumbar spine.After successful modeling,the model rabbits were subdivided into a control group,a model group,an ultrasonic needle-knife group,and a sham needle-knife group according to a random number table method,with six animals in each group.The ultrasonic needle-knife group underwent ultrasound-guided needle-knife release of the right yellow ligament of L7/S1,once every week,for a total of four times.The needle-knife approach in the sham needle-knife group was the same as that in the ultrasound needle-knife group,but the ligamentum flavum was not released.At 30 days after the intervention,MRI was used to observe the changes in the signal intensity of the nucleus pulposus within the L7/S1 segment.Hematoxylin-eosin staining was used to observe the morphological changes of the L7/S1 segment.Immunohistochemical staining was used to detect the expression of type I and II collagen in the nucleus pulposus of the L7/S1 segment.RT-PCR and western blot were used to detect the expression of integrin α5 and β1,p38,and nuclear factor κB in the L7/S1 segment. RESULTS AND CONCLUSION:MRI findings indicated that the nucleus pulposus of the intervertebral disc of rabbits in the model group was gray-black in color,and the gray value of the nucleus pulposus was significantly lower than that of the control group(P<0.01).The brightness of the nucleus pulposus of the intervertebral disc of the rabbits in the ultrasonic needle-knife group was elevated compared with that of the model group,and the gray value of the nucleus pulposus was higher than that of the model group(P<0.01).Results from hematoxylin-eosin staining showed that in the model group,the shape of the nucleus pulposus was irregular,the number of nucleus pulposus cells was reduced,the extracellular matrix was compressed,the fibrous ring was ruptured,the structure and boundary of the end plate were unclear,and the chondrocytes were arranged disorderly.Compared with the model group,the ultrasonic needle-knife group showed an increase in the number of the nucleus pulposus,an improvement in the rupture of the fibrous ring,and more regular arrangement of cartilage endplate cells.Results from immunohistochemical staining showed an increase in positive expression of type I collagen(P<0.01)and a decrease in positive expression of type II collagen in the nucleus pulposus of the model group compared with the control group as well as a decrease in positive expression of type I collagen and an increase in positive expression of type II collagen in the nucleus pulposus of the ultrasonic needle-knife group compared with the model group(P<0.01).RT-PCR and western blot assays showed that the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the model group were increased compared with that in the control group(P<0.01);the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the ultrasonic needle-knife group was decreased compared with that in the model group(P<0.01).To conclude,ultrasound-guided needle-knife release of the ligamentum flavum can improve the degree of lumbar disc degeneration in rabbits,which may be related to the inhibition of p38 and nuclear factor-κB expression by modulating integrin α5 and β1 expression.
8.Yijingtang Reduces Ovarian Inflammatory Responses in Rat Model of Diminished Ovarian Reserve via TLR4/MyD88/NF-κB Signaling Pathway
Heng HU ; Jijun CHU ; Zhe LI ; Haijing CHU ; Jing YU ; Chengcheng LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):20-30
ObjectiveTo investigate the effect and mechanism of Yijingtang (YJT) in treating diminished ovarian reserve (DOR) in rats by regulating the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. MethodsFifty female SD rats with normal estrous cycles were randomly allocated into blank, model, low- and high-dose (12.579 and 25.158 g·kg-1, respectively) YJT, and dehydroepiandrosterone (7.487 5 mg·kg-1) groups, with 10 rats in each group. The rats in other groups except the blank group were administrated with the tripterygium glycosides tablet suspension (5 mg·kg-1) by gavage for 14 days for the modeling of DOR. The rats in the drug treatment groups were administrated with corresponding drugs by gavage from day 15 for 30 consecutive days, and those in the blank and model groups received equal volumes of distilled water. The vaginal exfoliated cell smears were observed to assess the changes in the estrous cycle. The wet weight of bilateral ovaries was weighed for calculation of the ovarian index. Hematoxylin-eosin staining was performed to observe the histopathological changes in the ovaries and the proportions of follicles at various levels were calculated. The serum levels of sex hormones [follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and anti-Müllerian hormone (AMH)] and inflammatory factors [tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), and interleukin-10 (IL-10)] were determined by enzyme-linked immunosorbent assay. Real-time quantitative polymerase chain reaction(Real-time PCR) was conducted to determine the mRNA levels of TLR4, MyD88, NF-κB profilin α (IκBα), NF-κB and inflammatory factors in the ovarian tissue. Western blot was employed to measure the protein levels of factors related to the TLR4/MyD88/NF-κB signaling pathway in the ovarian tissue. Immunofluorescence (IF) was used to detect the nuclear translocation of NF-κB p65 in the ovarian tissue. ResultsCompared with the blank group, the model group showed disturbed estrous cycles, increased inflammatory infiltration in the ovarian tissue, decreases in ovarian index and proportion of presinusoidal follicles, and an increase in the proportion of atretic follicles (P<0.05, P<0.01). In addition, the model group showed elevated serum levels of FSH, LH, TNF-α, and IL-1β, up-regulated mRNA levels of TLR4, MyD88, IκBα, NF-κB, TNF-α, and IL-1β and protein levels of TLR4, MyD88, p-IκBα, and p-NF-κB p65 (P<0.01), lowered serum levels of AMH, E2, and IL-10, down-regulated mRNA level of IL-10 (P<0.01), and massive nuclear translocation of NF-κB p65 in the ovarian tissue. Compared with the model group, dehydroepiandrosterone and low and high doses of YJT restored the disturbed estrous cycle, reduced inflammatory infiltration in the ovarian tissue, increased the ovarian index (P<0.01), and changed the follicular composition ratio (P<0.01). Furthermore, the drugs lowered the serum levels of FSH, LH, TNF-α, and IL-1β, down-regulated the mRNA levels of TLR4, MyD88, IκBα, NF-κB, TNF-α, and IL-1β and the protein levels of TLR4, MyD88, p-IκBα, and p-NF-κB p65 (P<0.05, P<0.01), raised the serum levels of AMH, E2, and IL-10, up-regulated the mRNA level of IL-10 (P<0.05, P<0.01), and reduced the nuclear translocation of NF-κB p65 in the ovarian tissue. ConclusionYJT may inhibit the release and expression of inflammatory factors by regulating the TLR4/MyD88/NF-κB signaling pathway to attenuate the inflammatory responses in the ovarian tissue, thereby improving the ovarian function in DOR rats.
9.Yijingtang Reduces Ovarian Inflammatory Responses in Rat Model of Diminished Ovarian Reserve via TLR4/MyD88/NF-κB Signaling Pathway
Heng HU ; Jijun CHU ; Zhe LI ; Haijing CHU ; Jing YU ; Chengcheng LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):20-30
ObjectiveTo investigate the effect and mechanism of Yijingtang (YJT) in treating diminished ovarian reserve (DOR) in rats by regulating the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. MethodsFifty female SD rats with normal estrous cycles were randomly allocated into blank, model, low- and high-dose (12.579 and 25.158 g·kg-1, respectively) YJT, and dehydroepiandrosterone (7.487 5 mg·kg-1) groups, with 10 rats in each group. The rats in other groups except the blank group were administrated with the tripterygium glycosides tablet suspension (5 mg·kg-1) by gavage for 14 days for the modeling of DOR. The rats in the drug treatment groups were administrated with corresponding drugs by gavage from day 15 for 30 consecutive days, and those in the blank and model groups received equal volumes of distilled water. The vaginal exfoliated cell smears were observed to assess the changes in the estrous cycle. The wet weight of bilateral ovaries was weighed for calculation of the ovarian index. Hematoxylin-eosin staining was performed to observe the histopathological changes in the ovaries and the proportions of follicles at various levels were calculated. The serum levels of sex hormones [follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and anti-Müllerian hormone (AMH)] and inflammatory factors [tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), and interleukin-10 (IL-10)] were determined by enzyme-linked immunosorbent assay. Real-time quantitative polymerase chain reaction(Real-time PCR) was conducted to determine the mRNA levels of TLR4, MyD88, NF-κB profilin α (IκBα), NF-κB and inflammatory factors in the ovarian tissue. Western blot was employed to measure the protein levels of factors related to the TLR4/MyD88/NF-κB signaling pathway in the ovarian tissue. Immunofluorescence (IF) was used to detect the nuclear translocation of NF-κB p65 in the ovarian tissue. ResultsCompared with the blank group, the model group showed disturbed estrous cycles, increased inflammatory infiltration in the ovarian tissue, decreases in ovarian index and proportion of presinusoidal follicles, and an increase in the proportion of atretic follicles (P<0.05, P<0.01). In addition, the model group showed elevated serum levels of FSH, LH, TNF-α, and IL-1β, up-regulated mRNA levels of TLR4, MyD88, IκBα, NF-κB, TNF-α, and IL-1β and protein levels of TLR4, MyD88, p-IκBα, and p-NF-κB p65 (P<0.01), lowered serum levels of AMH, E2, and IL-10, down-regulated mRNA level of IL-10 (P<0.01), and massive nuclear translocation of NF-κB p65 in the ovarian tissue. Compared with the model group, dehydroepiandrosterone and low and high doses of YJT restored the disturbed estrous cycle, reduced inflammatory infiltration in the ovarian tissue, increased the ovarian index (P<0.01), and changed the follicular composition ratio (P<0.01). Furthermore, the drugs lowered the serum levels of FSH, LH, TNF-α, and IL-1β, down-regulated the mRNA levels of TLR4, MyD88, IκBα, NF-κB, TNF-α, and IL-1β and the protein levels of TLR4, MyD88, p-IκBα, and p-NF-κB p65 (P<0.05, P<0.01), raised the serum levels of AMH, E2, and IL-10, up-regulated the mRNA level of IL-10 (P<0.05, P<0.01), and reduced the nuclear translocation of NF-κB p65 in the ovarian tissue. ConclusionYJT may inhibit the release and expression of inflammatory factors by regulating the TLR4/MyD88/NF-κB signaling pathway to attenuate the inflammatory responses in the ovarian tissue, thereby improving the ovarian function in DOR rats.
10.Effect of CCNA2 on Prognosis of Colon Cancer by Regulating Immune Microenvironment of Tumor Cells
Peng YANG ; Ziyi QIU ; Lingling WANG ; Yuan HU ; Zhengzhen CHEN ; Meizhen ZHONG ; Feiyue YU ; Rongyuan QIU
Cancer Research on Prevention and Treatment 2025;52(4):305-312
Objective To investigate the relationship between cyclin A2 (CCNA2) and the prognosis of colon cancer, and its possible mechanism from the perspective of immune infiltration. Methods We downloaded the transcriptome data of colon cancer patients from The Cancer Genome Atlas database. Clinicopathological feature analysis and survival analysis were performed based on the expression levels of CCNA2. A total of 75 specimens of colon cancer and normal tissues were collected, and the expression level of CCNA2 was analyzed using immunohistochemical methods. Multivariate analysis was conducted to explore its relationship with clinicopathological features. Gene Set Enrichment Analysis (GSEA) was used to assess the potential molecular functions of CCNA2 in colon cancer. CIBERSORT algorithm was applied to calculate the correlation between CCNA2 and immune-cell infiltration in colon cancer. Results Database and immunohistochemical analyses indicated that CCNA2 was expressed at a significantly higher level in colon cancer tissues than normal tissues (P<0.001). The overall survival, disease-specific survival, and progression-free interval were all longer in the group with high CCNA2 expression than the group with low expression (all P<0.05). In tumor tissues, the expression level of CCNA2 decreased with increased pathological and TNM stages (P<0.05). The expression level of CCNA2 in normal tissues was consistently lower than that in colon cancer tissues across all clinical stages (all P<0.001). GSEA suggested that Wnt/β-catenin, KRAS, and other signaling pathways were enriched when CCNA2 was lowly expressed. CIBERSORT analysis revealed an increase in the infiltration of immune cells such as regulatory T cells and macrophages M0 when CCNA2 expression was low. Conclusion CCNA2 is highly expressed in colon cancer and closely associated with grade of pathology and TNM stage. It may recruit regulatory T cells through the KRAS and Wnt/β-catenin pathways, thereby reducing immune-cell infiltration and promoting colon cancer progression, leading to poor prognosis.

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