1.Effect of Qingfei Shenshi Decoction (清肺渗湿汤) Combined with Western Medicine on Clinical Effectiveness and Immune Function for Patients with Bronchial Asthma of Heat Wheezing Syndrome
Ying SUN ; Haibo HU ; Na LIU ; Fengchan WANG ; Jinbao ZONG ; Ping HAN ; Peng LI ; Guojing ZHAO ; Haoran WANG ; Xuechao LU
Journal of Traditional Chinese Medicine 2026;67(1):38-44
ObjectiveTo observe the clinical effectiveness and safety of Qingfei Shenshi Decoction (清肺渗湿汤) combined with western medicine for patients with bronchial asthma of heat wheezing syndrome, and to explore its potential mechanism of action. MethodsEighty-six participants with bronchial asthma of heat wheezing syndrome were randomly divided into treatment group and control group, each group with 43 participants. The control group received conventional western medicine, and the treatment group was additionally administered Qingfei Shenshi Decoction orally on the basis of the control group, 1 dose per day. Both groups were treated for 14 days. The primary outcome measure was clinical effectiveness; secondary outcome measures included traditional Chinese medicine (TCM) syndrome score, asthma control test (ACT) score, pulmonary function indices such as forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), peak expiratory flow (PEF), serum inflammatory factor levels including interleukin-4 (IL-4), tumour necrosis factor-α (TNF-α), and high-sensitivity C-reactive protein (hs-CRP), and immune function indices including CD3+, CD4+, CD8+, CD4+/CD8+. All outcome measures were evaluated before and after treatment. Vital signs were monitored, and electrocardiography, blood routine, urine routine, liver function, and renal function tests were performed before and after treatment. Adverse events and reactions during the study were recorded. ResultsA total of 80 patients completed the trial with 40 in each group. The total clinical effective rate of the treatment group was 97.5% (39/40), which was significantly higher than that of the control group (85.0%, 34/40, P<0.05). After treatment, both groups showed decreased TCM syndrome scores, IL-4, TNF-α, hs-CRP, and CD8+ levels, as well as increased ACT scores, CD3+, CD4+, CD4+/CD8+, FEV1, FVC, and PEF levels (P<0.05 or P<0.01). Moreover, the improvements in these indices were more significant in the treatment group than in the control group (P<0.05 or P<0.01). No significant abnormalities in safety indicators were observed in either group, and no adverse events or reactions occurred. ConclusionQingfei Shenshi Decoction combined with conventional western medicine for patients with bronchial asthma of heat wheezing syndrome can effectively improve the clinical symptoms, pulmonary function, and clinical effectiveness, with good safety. Its mechanism may be related to reducing inflammatory factor levels and regulating T lymphocyte subsets to improve immune function.
2.Effect of Anmeidan in Ameliorating Neuronal Synaptic Structural and Functional Impairment in Aged Sleep Deprivation Model via EphA4/ephrinA3 Signaling Pathway
Junlu ZHANG ; Guangjing XIE ; Ping WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):36-45
ObjectiveTo investigate the effects of Anmeidan (AMD) on protein expression of the ephrin type-A receptor 4 (EphA4)/ephrinA3 signaling pathway and synaptic structural function in an aged sleep deprivation model. MethodsSeventy-two 18-month-old aged mice were randomly divided into a blank group, a model group, AMD high-, medium-, and low-dose groups (26.26, 13.13, 6.565 g·kg-1·d-1, respectively), and a melatonin group (1.3 mg·kg-1·d-1), with 12 mice in each group. Cognitive function was assessed using the novel object recognition test. Hematoxylin-eosin (HE) staining was used to observe cell number and morphology in hippocampal tissues, and Nissl staining was performed to examine cellular structure and quantify Nissl bodies. Transmission electron microscopy was used to observe synaptic ultrastructure, with emphasis on changes in synaptic morphology and structure. Western blot was employed to detect the expression levels of EphA4, ephrinA3, brain-derived neurotrophic factor (BDNF), glutamate aspartate transporter (GLAST), glutamate transporter-1 (GLT-1), growth-associated protein 43 (GAP43), postsynaptic density protein 95 (PSD95), and synaptophysin (SYN) in hippocampal tissues. Immunofluorescence double labeling was performed to co-stain EphA4 and ephrinA3 with glial fibrillary acidic protein (GFAP) and neuronal nuclei antigen (NeuN), respectively, to observe the colocalization of target proteins with neurons and astrocytes. ResultsCompared with the blank group, the model group exhibited increased exploration time of familiar objects (P<0.01), while exploration time of novel objects and the recognition index were decreased (P<0.01). The number of neurons in the CA1, CA3, and dentate gyrus (DG) regions of the hippocampus was reduced, Nissl bodies were decreased, and synaptic structures were damaged. Protein expression levels of BDNF, GLAST, GLT-1, GAP43, PSD95, and SYN in hippocampal tissues were decreased, whereas the expression levels of EphA4, ephrinA3, and GFAP were increased. Compared with the model group, the AMD low-, medium-, and high-dose groups and the melatonin group showed increased exploration time of novel objects and higher novel object recognition indices (P<0.01), along with significantly reduced exploration time of familiar objects (P<0.01). Neuronal damage in the CA1 and DG regions was ameliorated, the number of Nissl bodies in the CA1 region was increased, and organelle and synaptic structural damage was alleviated. Protein expression levels of BDNF, GLAST, GLT-1, GAP43, PSD95, and SYN were increased, and protein expression levels of EphA4, ephrinA3, and GFAP were decreased (P<0.05,P<0.01). ConclusionAMD can regulate protein expression of the EphA4/ephrinA3 signaling pathway in an aged sleep deprivation model, enhance synaptic protein expression, and improve neuronal synaptic damage.
3.Perioperative immune dynamics and clinical outcomes in patients undergoing on-pump cardiac surgery
Zhiyuan CHENG ; Xinyi LIAO ; Juan WU ; Ping YANG ; Tingting WANG ; Qinjuan WU ; Wentong MENG ; Zongcheng TANG ; Jiayi SUN ; Jia TAN ; Jing LIN ; Dan LUO ; Hao WANG ; Chaonan LIU ; Jiyue XIONG ; Liqin LING ; Jing ZHOU ; Lei DU
Chinese Journal of Blood Transfusion 2026;39(1):31-43
Objective: To characterize perioperative dynamic changes in immune-cell phenotypes and inflammatory cytokines in patients undergoing CPB (cardiopulmonary bypass) cardiac surgery, and to explore their associations with postoperative outcomes. Methods: In this prospective cohort study, 120 adult patients who underwent elective cardiac surgery under CPB at West China Hospital from May 2022 to March 2023 were enrolled. Perioperative immune-cell phenotypes and concentrations of 40 inflammation-related cytokines were measured. The primary outcomes were the sequential organ failure assessment (SOFA) score at 24 h after surgery and ΔSOFA (the peak SOFA score within 48 h after surgery minus the preoperative SOFA score). Secondary outcomes included major adverse cardiovascular events (MACE), acute kidney injury (AKI), respiratory failure, severe liver injury, and infection. Results: The mean age of enrolled patients was 57±10 years. Of these, 52% (62/120) were male and 90% (108/120) underwent valve surgery. During the rewarming to the end of CPB, neutrophil counts rapidly increased (7.39×10
/L vs preoperative 3.07×10
/L, P<0.001), with significant upregulation of CD11b (7.30×10
/L vs preoperative 3.05×10
/L, P<0.001) and CD54 (7.15×10
/L vs preoperative 2.99×10
/L, P<0.001). Lymphocyte counts increased at the end of CPB (1.75×10
/L vs preoperative 1.12×10
/L, P<0.001) but decreased significantly at 24 h after surgery (0.59×10
/L vs preoperative 1.12×10
/L, P<0.001). Plasma analysis showed that multiple pro-inflammatory cytokines increased during CPB and remained elevated up to 24 h after surgery; five chemokines and the anti-inflammatory cytokine IL-10 peaked at the end of CPB. The SOFA score increased from 1 (1, 2) preoperatively to 7 (5, 10) at 24 h after surgery, with a ΔSOFA of 6 (4, 8). Within 30 days after surgery, 48 patients (40.0%) developed AKI, 17 (14.2%) developed infection, 4 (3.3%) developed severe liver injury, 3 (2.5%) developed respiratory failure, and 3 (2.5%) experienced MACE. During the 2-year follow-up, 8 patients (6.7%) experienced MACE and 5 (4.2%) died. Conclusion: Multi-organ dysfunction is common after cardiac surgery under CPB (median ΔSOFA, 6), accompanied by perioperative activation of multiple immune-cell subsets and upregulation of pro-inflammatory, anti-inflammatory, and chemotactic mediators. This study provides data-driven evidence and research clues for further investigation of the associations between CPB-related immune perturbations and postoperative organ dysfunction and clinical outcomes.
4.Current Status and Prospects of Research on Traditional Chinese Medicine Prevention and Treatment for Gastric Precancerous Lesions
Haiyan BAI ; Tai ZHANG ; Ping WANG ; Lin LIU ; Weichao XU ; Yaxin TIAN ; Lanshuo HU ; Qian YANG ; Xudong TANG
Journal of Traditional Chinese Medicine 2026;67(4):410-415
Traditional Chinese medicine (TCM), through its multi-target and systematic regulatory effects, has demonstrated unique advantages in the treatment of gastric precancerous lesions (GPL). At present, TCM theoretical research on GPL is mainly reflected in three aspects, the integration of macroscopic syndrome differentiation, the inflammation-carcinoma transformation mechanism, as well as the systematization and scientization of theoretical inheritance from famous TCM practitioners. High-quality evidence-based research findings serve as the foundation for clinical practice guidelines on GPL, and TCM has gained international academic recognition in the field of GPL prevention and treatment. Research on TCM mechanisms has yielded a series of important outcomes in the aspects of signaling pathways, gene expression regulation, cellular epigenetics, histone modification, and intestinal microecology. It is proposed that future research on GPL should focus on four key directions, establishing multi-omics data, exploring targeted intervention strategies on key regulatory nodes, advancing the standardization process of integrated traditional Chinese and western medicine prevention and treatment technologies, and constructing stratified screening and intervention platforms. The in-depth integration of TCM microcosmic mechanism of action with its macroscopic syndrome differentiation and treatment system, coupled with interdisciplinary research, will provide valuable references for the clinical treatment and scientific research of GPL.
5.Exosomes Treat Ischemic Stroke by Regulation of Ferroptosis Through the NRF2/SLC7A11/GPX4 Pathway in Mice
Yingtao XU ; Mengmeng WANG ; Ping LIN ; Haitao CHI ; Yi WANG ; Ying BAI
Laboratory Animal and Comparative Medicine 2026;46(1):20-31
ObjectiveA middle cerebral artery occlusion (MCAO) mouse model is established by electrocoagulation of the middle cerebral artery. The study examines the mechanism by which exosomes (EXO) derived from human amniotic mesenchymal stem cells (hAMSCs) improve ischemic stroke and regulate neural ferroptosis-related injury. MethodsThirty-two SPF-grade male C57BL/6J mice aged 6 - 8 weeks were randomly divided into four groups (n=8 per group): sham group (Sham), model group (MCAO), MCAO plus normal saline group (MCAO+NaCl), and MCAO plus exosome group (MCAO+EXO). The mouse MCAO model was established by electrocoagulation of the middle cerebral artery. Mice in the Sham group underwent exposure of the middle cerebral artery without electrocoagulation. Twenty-four hours before MCAO induction, mice in the MCAO+EXO group received a tail vein injection of 100 μL of exosomes derived from the culture supernatant of hAMSCs at a concentration of 9.5×1011 particles/mL. Mice in the MCAO+NaCl group were injected with an equal volume of normal saline via the tail vein. Twenty-four hours after model establishment, neurological deficits were evaluated using the Longa neurological deficit scoring system. Cerebral infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin and eosin (HE) staining was performed to evaluate morphological changes of neurons in the ischemic brain regions. The contents of ferrous iron (Fe2+), malondialdehyde (MDA), total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) in the infarct core and peri-infarct regions were determined using microcolorimetric assays to evaluate differences among groups. The mRNA expression levels of ferroptosis-related factors, including nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in the infarct core and peri-infarct regions were measured by real-time quantitative PCR. Protein expression levels of NRF2, SLC7A11, and GPX4 in the infarct and peri-infarct regions of each group were analyzed by Western blotting. ResultsCompared with the MCAO group, the Longa neurological deficit score was significantly reduced in the MCAO+EXO group (P<0.01). Prominent cerebral infarction was observed in the MCAO group, whereas the infarct volume ratio was markedly decreased in the MCAO+EXO group compared with the MCAO group (P<0.001). Histopathological analysis revealed that mice in the MCAO group exhibited obvious neuronal damage, including cytoplasmic vacuolar degeneration, nuclear pyknosis and fragmentation, unclear nuclear structure, and disorganized neuronal arrangement, compared with the Sham group. In contrast, neurons in the MCAO+EXO group showed relatively preserved morphology, with intact cellular structures and large, regular nuclei located centrally within the cells. Biochemical analysis demonstrated that Fe2+ and MDA levels in the infarct core and peri-infarct regions were significantly increased in the MCAO group compared with the Sham group (P<0.001). These levels were significantly reduced in the MCAO+EXO group compared with the MCAO group (P<0.01). In addition, total glutathione (total GSH), oxidized glutathione (GSSG), and reduced glutathione (GSH) levels were markedly decreased in the MCAO group relative to the Sham group (P<0.01). Compared with the MCAO group, the MCAO+EXO group exhibited significantly increased levels of total GSH and GSH (P<0.001), while no significant change was observed in GSSG levels (P>0.05). Furthermore, both mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2 (NRF2), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) were significantly downregulated in the MCAO group compared with the Sham group (P<0.01, P<0.001). In contrast, both mRNA and protein expression levels of NRF2, SLC7A11, and GPX4 were significantly upregulated in the MCAO+EXO group compared with the MCAO group (P<0.05). ConclusionIn the mouse MCAO model, tail vein injection of exosomes derived from hAMSCs can improve motor function, reduce infarct area, protect neuronal cell morphology, and reduce the degree of nerve injury. Exosomes may exert a protective effect by activating the NRF2/SLC7A11/GPX4 pathway and reducing ferroptosis in neuronal cells of MCAO model mice.
6.Clinical analysis of assisted reproductive technology assisted pregnancy outcome in female patients with thyroid cancer after surgery
Xiang YAO ; Wenjuan XU ; Jianye WANG ; Qun GAO ; Gang ZHAO ; Ping ZHOU
Acta Universitatis Medicinalis Anhui 2026;61(1):151-155
ObjectiveTo evaluate the pregnancy outcomes of assisted reproductive technology (ART) in women with a history of thyroid cancer who retained fertility intentions after completing cancer treatment. MethodsA retrospective analysis was performed on 61 patients with a history of thyroid cancer who underwent in vitro fertilization/intracytoplasmic sperm microinjection and embryo transfer (IVF/ICSI-ET). These patients were included as the case group. A total of 122 non-cancer patients who received ART during the same period were selected as the control group using 1∶2 matching based on age and oocyte retrieval time. Baseline characteristics, outcomes of the first ART cycle, and cumulative pregnancy outcomes were compared between the two groups. ResultsThere was no significant difference in the basic data, the total amount of gonadotropin (Gn) and the days of use between the case group and the control group (P>0.05). However, the case group had significantly fewer retrieved oocytes, mature oocytes (MII), lower fertilization and cleavage rates, and fewer transferable and high-quality embryos, as well as fewer embryos transferred during the first cycle (P < 0.05). However, there was no significant difference in the rate of first embryo implantation and first clinical pregnancy between the two groups (P>0.05). In the analysis of cumulative outcomes, the two groups did not show statistically significant differences in the cumulative pregnancy rate, clinical pregnancy rate per transfer cycle, the number of oocyte retrieval cycles required per live birth, the number of embryo transfer cycles required per live birth, and the number of embryos used for each live birth (P>0.05). However, the cumulative live birth rate was significantly lower in the case group compared to the control group (P=0.005). ConclusionAfter treatment for thyroid cancer, when ART is used to help pregnant women, the pregnancy outcome is comparable to that of women without tumors. Individualized reproductive management and timely fertility preservation strategies are recommended to optimize reproductive outcomes in this population.
7.Mechanism of Sancao Anshen Prescription in Regulating cAMP/PKA Signaling Pathway to Improve Sleep and Depressive States in Zebrafish with SD Models
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):180-190
ObjectiveBy establishing a light sleep deprivation (SD) model and using the traditional Chinese medicine (TCM) compound Sancao Anshen prescription for intervention, this study aims to observe one of the effects of Sancao Anshen prescription on the sleep and depressive states of zebrafish in the SD model and explore the mechanism of action of this drug in regulating the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling pathway. MethodsA total of 240 six-month-old wild-type AB zebrafish were randomly divided into a blank group, a model group, low-dose, medium-dose, and high-dose groups of Sancao Anshen prescription (0.28, 0.83, 2.48 g·L-1), and a melatonin group (0.2 g·L-1), with 40 fish in each group. Except for the blank group, all others were exposed to LED lights (150 lux) for three days to construct the sleep deprivation model, and were treated with the corresponding doses of Sancao Anshen decoction and melatonin solution for three days. 24 h movement behavior was used to detect diurnal movement trajectories. A T-shaped maze was employed to detect learning and memory functions, and a new tank experiment was conducted to detect depression-like behaviors in zebrafish. Hematoxylin-eosin (HE) staining was used to observe the morphology of hypothalamic neurons, and transmission electron microscopy was utilized to observe the ultrastructure of hypothalamic cells. Immunohistochemical (IHC) was used to measure the positive expression of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the hypothalamus, and enzyme-linked immunosorbent assay (ELISA) was employed to determine the content of cAMP, 5-hydroxytryptamine (5-HT), gamma-aminobutyric acid (GABA), and glutamic acid (Glu) in brain tissue. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure the expression of PKA, cAMP response element-binding (CREB), and brain-derived neurotrophic factor (BDNF) mRNAs and proteins and their phosphorylation levels (p-PKA, p-CREB) in brain tissue. ResultsCompared with those in the blank group, the resting time and resting count in the model group decreased significantly (P<0.01), while the gross motor time and gross motor count increased significantly (P<0.01). The latency time to enter the EC region in each administration group increased significantly (P<0.01). The exploration time towards the top and the number of times entering the top decreased significantly (P<0.01), and the incubation period of the first ascent increased significantly (P<0.01). The number of hypothalamic neurons decreased significantly, and the neurons exhibited irregular shapes, sparse arrangement, and nuclear condensation. Nuclear collapse, nuclear membrane rupture and dissolution, chromatin condensation, mitochondrial swelling and deformity, and plate-like cristae rupture or disappearance were observed. The positive expression of TNF-α and IL-1β in the hypothalamus was significantly decreased (P<0.01). The content of cAMP, GABA, and 5-HT in brain tissue was significantly downregulated, while the content of Glu was significantly upregulated (P<0.01). The mRNA of PKA, CREB, and BDNF was significantly downregulated (P<0.01), and the protein expression of p-PKA, p-PKA/PKA, p-CREB, p-CREB/CREB, and BDNF was significantly decreased (P<0.05, P<0.01). Compared with those in the model group, the resting time in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group increased (P<0.05, P<0.01), and the resting count in each administration group significantly increased (P<0.05, P<0.01). The gross motor time and gross motor count significantly decreased (P<0.05, P<0.01). The latency time to enter the EC region decreased (P<0.01), and the exploration time towards the top increased (P<0.01). The time for the first ascent in the high-dose group of Sancao Anshen prescription and the melatonin group was shortened (P<0.05, P<0.01), and the number of times entering the top increased (P<0.01). The morphology of neurons in each administration group improved, with the gap decreased, the nuclear membrane relatively intact, and mitochondrial swelling improved. The positive expression of IL-1β in each administration group significantly decreased (P<0.01). The positive expression of TNF-α in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group significantly decreased (P<0.01). The Glu content in the low-dose group of Sancao Anshen prescription decreased, while cAMP and GABA levels increased (P<0.05, P<0.01). There was no statistically significant difference in 5-HT content. The medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group all showed significant increases in cAMP, 5-HT, and GABA levels (P<0.01), and a significant decrease in Glu content (P<0.01). The mRNA of CREB in the low-dose group of Sancao Anshen prescription was significantly upregulated (P<0.01), while there was no statistically significant difference in the mRNA of PKA and BDNF. The mRNA of PKA, CREB, and BDNF in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group were all significantly upregulated (P<0.01). The protein expression of p-PKA, p-PKA/PKA, p-CREB, p-CREB/CREB, and BDNF in the high-dose group of Sancao Anshen prescription and the melatonin group increased significantly (P<0.05, P<0.01), and the protein expression of p-PKA, p-CREB, and BDNF in the medium-dose group of Sancao Anshen prescription increased (P<0.05). ConclusionThe improvement of sleep and depressive states in zebrafish with the SD model by Sancao Anshen prescription may be related to the inhibition of inflammatory factors such as TNF-α and IL-1β, the reduction in Glu, and the elevation in the content of neurotransmitters such as GABA and 5-HT via the cAMP/PKA signaling pathway.
8.Influencing factors for poor prognosis of drug-induced liver injury in Chinese population: a systematic review
Weimei WANG ; Lidan WANG ; Jia MENG ; Ze PING ; Xiaoyan ZHANG
China Pharmacy 2026;37(5):665-669
OBJECTIVE To systematically evaluate the influencing factors affecting the poor prognosis of drug-induced liver injury (DILI) in the Chinese population, and to provide evidence-based support for early identification and interventions of DILI. METHODS Retrieved from PubMed, Medline, Embase, the Cochrane Library, CNKI, Wanfang database, China biomedical medicine database (CBM) and VIP, clinical studies (case-control studies, cohort studies) related to influencing factors for poor prognosis of DILI were collected from inception to May 31, 2025. After literature screening, data extraction and quality evaluation of included studies, meta-analysis was carried out by using RevMan 5.4 software. RESULTS A total of 17 literature were included, involving 4 078 DILI patients, of whom 673 were in the poor prognosis group and 3 405 were in the favorable prognosis group. Meta-analysis showed that history of liver disease (OR=2.47, 95%CI was 1.61-3.78, P <0.001), alcohol drinking history (OR=1.77, 95%CI was 1.22-2.56, P =0.003), Chinese herbal medicine/Chinese patent medicine (OR=1.87, 95%CI was 1.30-2.70, P <0.001), non-hepatocellular injury type (OR=1.70, 95%CI was 1.37-2.10, P <0.001), international normalized ratio (INR) elevated (OR=2.51, 95%CI was 1.97-3.19, P <0.001), and alanine transamine (ALT) elevated (OR=1.27, 95%CI was 1.14-1.41, P <0.001) were risk factors of poor prognosis in DILI. Higher albumin (ALB) level (OR=0.47, 95%CI was 0.39-0.57, P <0.001), elevated prothrombin activity (PTA) (OR=0.88, 95%CI was 0.85-0.91, P <0.001) and more than 2 kinds of hepatoprotective drugs (OR=0.62, 95%CI was 0.41-0.95, P =0.030) were protective factors for poor prognosis of DILI. CONCLUSIONS Patients with alcohol drinking history, history of liver disease, elevated INR, elevated ALT, taking Chinese herbal medicine/Chinese patent medicine, and non-hepatocellular injury type of DILI have a greater risk of poor prognosis, and higher ALB level, higher PTA and more than 2 kinds of hepatoprotective drugs can reduce the risk of poor prognosis of DILI.
9.A Case Study of Using Assisted Reproductive Technology to Rescue Genetically Modified Mice with Reproductive Disorder Phenotypes
Qianqian WANG ; Sijue TAO ; Zhen WEI ; Huihui JIN ; Ping LIU ; Lie WANG
Laboratory Animal and Comparative Medicine 2025;45(1):79-86
ObjectiveThe utilization of assisted reproductive technology to rescue genetically modified mouse strains with reproductive disorders provides a reference for improving techniques to preserve valuable experimental mouse strains. MethodsIn vitro fertilization-embryo transfer (IVF-ET) technology was performed on 28 strains of infertile male mice aged 9-18 months. Several indicators such as sperm density and sperm motility in infertile male mice were assessed to select the most viable sperm for IVF-ET experiments. Fertility rate, abnormal egg rate, and birth rate were recorded after the birth of the pups. An optimized ovarian transplantation procedure was applied to 12 strains of infertile female mice aged 8-18 months. 6-week-old female mice with the same genetic background were selected as recipients. One intact ovary was removed from each recipient mouse, and the contralateral oviduct was ligated. An ovary from a donor mouse was isolated and transplanted orthotopically into the side where the ovary had been removed in the recipient mouse. Twenty-one days post-surgery, recipient mice were co-housed with 8-week-old wild type male mice of the same genetic background for breeding. Data such as the pregnancy rate and live birth rate of the recipients were recorded after the birth of the pups. ResultsIVF-ET successfully rescued 28 mouse strains, with the oldest male mice being 18 months old. The success rate of the first round of IVF-ET experiments was 89.29% (25/28). The average fertility rate of IVF in infertile male mice was (51.01±14.97)%, the abnormal egg rate was (9.03±5.28)%, and the birth rate of offspring mice was (18.60±7.03)%. 39 out of 40 ovarian transplant recipient mice survived, with a pregnancy rate of 33.33% (13/39) for ovarian transplant recipients, and a live birth rate of 17.95% (7/39). Four mouse strains were successfully rescued using optimized ovarian transplantation technology, with the oldest female mice being 18 months old. 8 strains were not rescued as they failed to produce offspring that survived to sexual maturity. ConclusionIVF-ET is an effective approach for rescuing mice with reproductive disorders caused by different reasons, especially for those beyond the optimal breeding age. Ovarian transplantation technology can also be used as an alternative for aged female mice. But its success rate is relatively lower than that of IVF-ET, and carries a higher experimental risk.
10.Effects of different exercise interventions on carboxylesterase 1 and inflammatory factors in skeletal muscle of type 2 diabetic rats
Shujuan HU ; Ping CHENG ; Xiao ZHANG ; Yiting DING ; Xuan LIU ; Rui PU ; Xianwang WANG
Chinese Journal of Tissue Engineering Research 2025;29(2):269-278
BACKGROUND:Carboxylesterase 1 and inflammatory factors play a crucial role in regulating lipid metabolism and glucose homeostasis.However,the effects of different exercise intensity interventions on carboxylesterase 1 and inflammatory factors in skeletal muscle of type 2 diabetic rats remain to be revealed. OBJECTIVE:To investigate the effects of different exercise intensity interventions on carboxylesterase 1 and inflammatory factors in skeletal muscle of type 2 diabetic rats. METHODS:Thirty-two 8-week-old male Sprague-Dawley rats were randomly divided into normal control group(n=12)and modeling group(n=20)after 1 week of adaptive feeding.Rat models of type 2 diabetes mellitus were prepared by high-fat diet and single injection of streptozotocin.After successful modeling,the rats were randomly divided into diabetic control group(n=6),moderate-intensity exercise group(n=6)and high-intensity intermittent exercise group(n=6).The latter two groups were subjected to treadmill training at corresponding intensities,once a day,50 minutes each,and 5 days per week.Exercise intervention in each group was carried out for 6 weeks.After the intervention,ELISA was used to detect blood glucose and blood lipids of rats.The morphological changes of skeletal muscle were observed by hematoxylin-eosin staining.The mRNA expression levels of carboxylesterase 1 and inflammatory cytokines were detected by real-time quantitative PCR.The protein expression levels of carboxylesterase 1 and inflammatory cytokines were detected by western blot and immunofluorescence. RESULTS AND CONCLUSION:Compared with the normal control group,fasting blood glucose,triglyceride,low-density lipoprotein cholesterol,insulin resistance index in the diabetic control group were significantly increased(P<0.01),insulin activity was decreased(P<0.05),and the mRNA and protein levels of carboxylesterase 1,never in mitosis gene A related kinase 7(NEK7)and interleukin 18 in skeletal muscle tissue were upregulated(P<0.05).Compared with the diabetic control group,fasting blood glucose,triglyceride,low-density lipoprotein cholesterol,and insulin resistance index in the moderate-intensity exercise group and high-intensity intermittent exercise group were down-regulated(P<0.05),and insulin activity was increased(P<0.05).Moreover,compared with the diabetic control group,the mRNA level of NEK7 and the protein levels of carboxylesterase 1,NEK7 and interleukin 18 in skeletal muscle were decreased in the moderate-intensity exercise group(P<0.05),while the mRNA levels of carboxylesterase 1,NEK7,NOD-like receptor heat protein domain associated protein 3 and interleukin 18 and the protein levels of carboxylesterase 1 and interleukin 18 in skeletal muscle were downregulated in the high-intensity intermittent exercise group(P<0.05).Hematoxylin-eosin staining showed that compared with the diabetic control group,the cavities of myofibers in the moderate-intensity exercise group became smaller,the number of internal cavities was reduced,and the cellular structure tended to be more intact;the myocytes of rats in the high-intensity intermittent exercise group were loosely arranged,with irregular tissue shape and increased cavities in myofibers.To conclude,both moderate-intensity exercise and high-intensity intermittent exercise can reduce blood glucose,lipid,insulin resistance and carboxylesterase 1 levels in type 2 diabetic rats.Moderate-intensity exercise can significantly reduce the expression level of NEK7 protein in skeletal muscle,while high-intensity intermittent exercise can significantly reduce the expression level of interleukin 18 protein in skeletal muscle.In addition,the level of carboxylesterase 1 is closely related to the levels of NEK7 and interleukin 18.

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