ObjectiveTaking the cuproptosis/oxidative stress pathway as the entry point， this study investigated the effect and mechanism of Liangyi Paste on hepatic lipid deposition in naturally aged mice fed with a high-fat diet. MethodsAfter adaptive feeding， 80 ten-week-old male C57BL/6 mice were used. Thirty of them were randomly divided into three groups （10 mice per group）： The 12-month-old control group （12MCON）， the 15-month-old control group （15MCON）， and the 15-month-old group with a high-fat diet （15MHFD）. The 12MCON and 15MCON groups were continuously fed a standard diet， while the 15MHFD group started receiving a high-fat diet at 12 months of age. Tissue samples were collected at the corresponding time points for each group. The remaining 50 mice were randomly divided into five groups （10 mice per group）： the 20-month-old control group （20MCON）， the model group， and the low-， medium-， and high-dose Liangyi Paste groups （2.91 ， 5.82 ， 11.64 g·kg^-1·d^-1， respectively）. The 20MCON group was continuously fed a standard diet， while the other groups started receiving a high-fat diet at 15 months of age. At 18 months of age， the Liangyi Paste groups were administered the corresponding doses of Liangyi Paste by gavage， while the 20MCON and model groups were given an equal volume of saline by gavage. After 8 weeks of continuous gavage （when the mice reached 20 months of age）， tissue samples were collected. Hepatic TG levels were measured using assay kits； liver histology and lipid deposition were observed via hematoxylin-eosin （HE） and oil red O staining； reactive oxygen species （ROS） were detected by enzyme-linked immunosorbent assay （ELISA）； Cu²⁺， superoxide dismutase （SOD）， and malondialdehyde （MDA） levels were measured by colorimetry； mRNA and protein expression of genes related to cuproptosis and oxidative stress pathways were analyzed by Real-time polymerase chain reaction（Real-time PCR） and Wes automated protein expression system. ResultsCompared with 12MCON， the 15MCON group showed significantly increased hepatic TG， Cu²⁺， ROS， and MDA levels （P<0.01）， decreased SOD （P<0.01）， hepatocyte swelling， and disordered arrangement. The mRNA and protein levels of ferredoxin 1 （FDX1）， dihydrolipoamide S-acetyltransferase （DLAT）， heat shock protein 70 （HSP70）， dihydrolipoamide dehydrogenase （DLD）， pyruvate dehydrogenase E1 subunit-β （PDHB）， nuclear factor erythroid 2-related factor 2 （Nrf2）， and peroxisome proliferator-activated receptor γ （PPARγ） were significantly elevated （P<0.05， P<0.01）. Compared with 15MCON group， the 15MHFD and 20MCON groups exhibited further increases in TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， and aggravated hepatocyte swelling and disorder. There were increased lipid droplets with mild vacuolization in the 15MHFD group， and no significant lipid deposition was observed in the 20MCON group. FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and protein levels were significantly increased （P<0.05， P<0.01）. Compared with 20MCON group， the model group demonstrated markedly elevated TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， severe hepatic steatosis， and upregulated expression of FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and proteins （P<0.05， P<0.01）. All abnormalities were significantly reversed after Liangyi Paste treatment. ConclusionLiangyi paste can ameliorate hepatic lipid deposition in naturally aged mice with a high-fat diet by modulating the cuproptosis/oxidative stress pathway.

Polycystic ovary syndrome (PCOS) is the predominant cause of subfertility in reproductive-aged women; however, its pathophysiology remains unknown. Neurotensin (NTS) is a member of the gut-brain peptide family and is involved in ovulation; its relationship with PCOS is unclear. Here, we found that NTS expression in ovarian granulosa cells and follicular fluids was markedly decreased in patients with PCOS. In the in vitro culture of cumulus-oocyte complexes, the neurotensin receptor 1 (NTSR1) antagonist SR48692 blocked cumulus expansion and oocyte meiotic maturation by inhibiting metabolic cooperation and damaging the mitochondrial structure in oocytes and surrounding cumulus cells. Furthermore, the ERK1/2-early growth response 1 pathway was found to be a key downstream mediator of NTS/NTSR1 in the ovulatory process. Animal studies showed that in vivo injection of SR48692 in mice reduced ovulation efficiency and contributed to irregular estrus cycles and polycystic ovary morphology. By contrast, NTS partially ameliorated the ovarian abnormalities in mice with dehydroepiandrosterone-induced PCOS. Our findings highlighted the critical role of NTS reduction and consequent abnormal NTSR1 signaling in the ovulatory dysfunction of PCOS, suggesting a potential strategy for PCOS treatment.
Polycystic Ovary Syndrome/physiopathology* ; Female ; Animals ; Neurotensin/metabolism* ; Receptors, Neurotensin/antagonists & inhibitors* ; Mice ; Ovulation/drug effects* ; Humans ; Granulosa Cells/metabolism* ; Adult ; Oocytes/metabolism* ; MAP Kinase Signaling System ; Signal Transduction ; Follicular Fluid/metabolism* ; Disease Models, Animal ; Gonadotropin-Releasing Hormone/analogs & derivatives*

Abstract： ObjectiveThe study aims to explore the effects and regulatory roles of glucose transporter 1 （GLUT1） on the proliferation， migration， adhesion， and angiogenesis of human umbilical vein endothelial cells （HUVECs） under ischemia-hypoxic conditions. MethodsIn vitro experiments were conducted to subject HUVECs to an ischemia-hypoxic-mimicking environment （1% O₂， 5% CO₂， 94% N₂）. The biological characteristics of HUVECs under normoxic and ischemia-hypoxic conditions were compared by assessing cell viability， proliferation capacity， and examining the expression changes of GLUT1， HIF-1α， and VEGFA proteins under ischemia-hypoxia using Western blot technology. Further， GLUT1 was overexpressed using plasmid transfection and the proliferation， migration， adhesion， and angiogenic capabilities of HUVECs were evaluated through scratch assays， cell adhesion assays， and tube formation assays. Mitochondrial morphological changes were observed by transmission electron microscopy，and oxygen consumption rate （OCR） was detected by Seahorse metabolic analyzer to evaluate mitochondrial function. ResultsCompared with normoxic conditions， the ischemia-hypoxic environment significantly inhibited the proliferation， cell viability， migration， and adhesion capabilities of HUVECs and impaired their angiogenic potential. The expression levels of GLUT1， HIF-1α and VEGFA proteins were also markedly reduced. However， when GLUT1 expression was upregulated， the migration， adhesion， and angiogenic capabilities of HUVECs were significantly improved， and the protein expression levels of HIF-1α， VEGFA and VEGFR were increased. Transmission electron microscopy revealed that ischemic-hypoxia leads to mitochondrial swelling and matrix damage， while GLUT1 overexpression significantly alleviates mitochondrial morphology abnormalities. OCR results suggest that GLUT1 overexpression may enhance oxidative phosphorylation of endothelial cells in ischemic-hypoxic environments to improve energy metabolism. These results suggest that GLUT1 may influence the function and angiogenic potential of HUVECs by regulating glucose metabolism and energy supply. ConclusionsThis study reveals the significant regulatory role of GLUT1 in the function of HUVECs under ischemia-hypoxic conditions， potentially through modulating cellular energy metabolism and signal transduction pathways， thereby affecting cell proliferation， migration， adhesion， and angiogenesis. These findings provide a new perspective on the role of GLUT1 in cardiovascular diseases and may offer potential targets for the development of new therapeutic strategies.

Objective:To explore the anal function and postoperative complications of 2-stage Turnbull-Cutait pull-through coloanal anastomosis (TCA) for low rectal cancer.Methods:Patients undergoing radical rectal cancer resection from Feb 2023 to Nov 2024 in the First Hospital of Jilin University were divided into the TCA surgery group and the low anterior resection combined with prophylactic stoma (LAR) surgery group.Results:Among the 102 patients, there were 50 cases in the TCA group and 52 cases in the LAR group. In the single-arm analysis of the TCA group, the overall complication rate was 44%. The incidence rates of severe LARS at 1 month, 3 months, and 6 months after surgery were 97%, 77%, and 64% respectively. There was no significant difference in the complication rate within 30 days after surgery between the two groups,(44% vs. 38%, χ2=0.135, P>0.05). There was no significant difference in the incidence rate of severe LARS between the TCA group and the LAR group (77% vs. 69%, χ2=0.202, P>0.05), and there was not significant difference in the incidence rate of severe LARS between the two groups at the 6th month after surgery,(64% vs. 48%, χ2=1.132, P>0.05). Conclusion:In patients who underwent TCA surgery, the LARS symptoms gradually decreased over time. Compared with patients undergoing low anterior resection and stoma reversal, there were no significant differences in complications within 30 days after surgery and LARS symptoms within half a year.

Objective:To explore the value of serum soluble E-cadherin (sEC) and cyclooxygenase-2 (COX-2) levels in predicting postoperative recurrence and metastasis of breast cancer.Methods:198 female breast cancer patients who underwent surgical treatment in Huai’an Hospital Affiliated to Xuzhou Medical University (Huai’an Second People’s Hospital) From Mar. 2023 to Mar. 2025 were selected and followed up for 3 years. The recurrence and metastasis were counted. Patients with recurrence and metastasis were included in the poor prognosis group, and patients without recurrence and metastasis were included in the good prognosis group. The general data, preoperative serum sEC and COX-2 levels were compared between the two groups. The influencing factors of recurrence and metastasis of breast cancer after operation were analyzed. The dose-response relationship and predictive value of serum sEC and COX-2 levels with recurrence and metastasis were analyzed. Logistic regression equation (LR) model 1 was constructed according to conventional influencing factors, and LR model 2 was constructed according to conventional influencing factors combined with serum sEC and COX-2. The predictive efficacy and accuracy of the two models were analyzed by receiver operating characteristic curve (ROC) and calibration curve. According to the ratio of 7∶3, another 85 female breast cancer patients who underwent surgical treatment in Huai’an Hospital Affiliated to Xuzhou Medical University (Huai’an Second People’s Hospital) from May. 2023 to May. 2025 were selected for external validation (validation set) .Results:Among 198 breast cancer patients who underwent postoperative follow-up for 3 years, 3 cases were lost to follow-up, and the recurrence and metastasis rate among the 195 patients who completed the 3-year postoperative follow-up was 26.15% (51/195). In the good prognosis group, the proportion of TNM stage III was 20.14%, the proportion of lymph node metastasis was 11.81%, the preoperative serum carbohydrate antigen 153 (CA153) level was (54.19±10.84) U/mL, the carbohydrate antigen 125 (CA125) level was (46.03±10.27) U/mL, the sEC level was (1987.65±191.37) ng/mL, and the COX-2 level was (17.85±5.21) ng/mL. In the poor prognosis group, the proportion of TNM stage III was 45.10%, the proportion of lymph node metastasis was 35.29%, the preoperative serum CA153 level was (65.87±12.23) U/mL, the CA125 level was (57.76±11.51) U/mL, the sEC level was (2 295.37±261.48) ng/mL, and the COX-2 level was (10.42±3.16) ng/mL. Compared with the good prognosis group, they were increased ( t/χ 2=12.00, 14.11, 6.39, 6.79, 8.92, 12.00, P<0.05). Multivariate Logistic regression analysis showed that TNM stage III ( OR=2.078, 95%CI =1.569-2.751, P<0.05), lymph node metastasis ( OR=2.435, 95 %CI=1.843-3.216, P<0.05), serum CA153 ( OR= 1.180,95 %CI=1.026-1.357, P<0.05), CA125 ( OR=1.206,95 %CI=1.033-1.408, P<0.05), sEC ( OR=1.011,95 %CI= 1.007-1.015, P<0.05), COX-2 ( OR=1.378,95 %CI=1.128-1.683, P<0.05) were independent risk factors for postoperative recurrence and metastasis of breast cancer. Restricted cubic spline (RCS) analysis showed that serum sEC and COX-2 levels were positively correlated with the risk of recurrence and metastasis of breast cancer ( P<0.05). ROC analysis showed that serum sEC (AUC=0.762,95 %CI=0.696-0.820, P<0.05) and COX-2 (AUC=0.757,95 %CI=0.691-0.815, P<0.05) could be used as biological indicators to predict postoperative recurrence and metastasis of breast cancer. The AUC of LR model 1 in predicting postoperative recurrence and metastasis of breast cancer was 0.862 (95 %CI=0.805-0.907, P<0.05). The AUC of LR model 2 in predicting postoperative recurrence and metastasis of breast cancer was 0.965 (95 %CI= 0.891-0.983, P<0.05), which was significantly larger than that of LR model 1 ( Z=2.015, P<0.05). The calibration curve analysis showed that the calibration degree of LR model 1 in predicting postoperative recurrence and metastasis of breast cancer was good, and the prediction results were in good agreement with the actual observation results. The calibration degree of LR model 2 was high, and the prediction results were in good agreement with the actual observation results. The results of external validation showed that the sensitivity of LR model 2 in predicting postoperative recurrence and metastasis of breast cancer was 82.61%, the specificity was 93.55%, the accuracy was 90.59%, and the Kappa value was 0.762 (95 %CI: 0.549-0.974) . Conclusions:Preoperative serum sEC and COX-2 levels are independent risk factors for postoperative recurrence and metastasis of breast cancer. They can be used as biological indicators to predict postoperative recurrence and metastasis of breast cancer. Combined with conventional indicators, they can significantly improve the predictive efficacy.

Monoclonal antibodies(mAbs)represent the most extensively utilized class of therapeutic proteins in clinical practice.While the majority of currently marketed mAbs are primarily employed for the treatment of non-infectious diseases,such as tumors and autoimmune disorders,the number of monoclonal antibody drugs approved globally for the prevention or treatment of infectious diseases has been progressively increasing since the onset of the COVID-19 pandemic.On March 25,2023,the World Health Organization(WHO)published the"Guidelines on the Non-Clinical and Clinical Evaluation of Monoclonal Antibodies and Related Products Intended for the Prevention or Treatment of Infectious Diseases".However,China has yet to release analogous guidelines.This article aims to elucidate the non-clinical evaluation aspects of monoclonal antibody drugs for infectious disease prevention and treatment as outlined in the WHO guidelines,with a focus on regulatory considerations,overarching principles of non-clinical evaluation,as well as pharmacodynamics,pharmacokinetics,and toxicology studies.It is intended to serve as a reference for the non-clinical research and evaluation of such novel drugs in China.

OBJECTIVE To explore the expression levels of microribonucleic acid-21(miR-21)and helper T cell(Th)1/Th2 cytokines in peripheral blood of the asthma children complicated with respiratory tract infections of vi-ruses and analyze the significance.METHODS A total of 90 asthma children with respiratory tract infections of vi-ruses(the infection group)and 43 asthma children without respiratory tract infections of viruses(the no infection group)who were treated in the 904th Hospital of the Joint Logistics Support Force of The People's Liberation Ar-my of China were enrolled in the study,and the clinical data were collected from the enrolled patients.The distri-bution of viruses from the children of the infection group was analyzed.The levels of peripheral blood miR-21,Th1 cytokines[interferon-γ(IFN-γ),interleukin(IL)-2]and Th2 cytokines[IL-4,IL-5]were compared between the two groups.The children of the infection group were divided into the mild group,the moderate group and the severe group according to the severity of disease.The levels of the above indexes were compared among the chil-dren with the various degree of illness condition.The association of the illness condition with miR-21,IFN-γ,IL-2,IL-4 and IL-5 was analyzed.RESULTS Respiratory syncytial virus was dominant among the respiratory tract vi-ruses in the infection group,accounting for 32.22％.The levels of IFN-γ and IL-2 of the infection group were low-er than those of the no infection group(P＜0.05),the levels of miR-21,IL-4 and IL-5 of the infection group were higher than those of the no infection group(P＜0.05).The levels of IFN-γ and IL-2 of the severe group were lower than those of the mild group and the moderate group(P＜0.05),while the levels of miR-21,IL-4 and IL-5 of the severe group were higher than those of the mild group and the moderate group(P＜0.05);the levels of IFN-γ and IL-2 of the moderate group were lower than those of the mild group(P＜0.05),and the levels of miR-21,IL-4 andIL-5 of the moderate group were higher than those of the mild group(P＜0.05).Spearman correlation analysis showed that the illness condition of the children was positively correlated with miR-21,IL-4 and IL-5(P＜0.05),which was negatively correlated with IFN-γ and IL-2(P＜0.05).CONCLUSIONS The respiratory tract infections of viruses may aggravate the abnormal rise of peripheral blood miR-21 and the imbalance of Th1/Th2.The above indexes are closely associated with the illness condition of the children.

Objective:To explore the value of serum soluble E-cadherin (sEC) and cyclooxygenase-2 (COX-2) levels in predicting postoperative recurrence and metastasis of breast cancer.Methods:198 female breast cancer patients who underwent surgical treatment in Huai’an Hospital Affiliated to Xuzhou Medical University (Huai’an Second People’s Hospital) From Mar. 2023 to Mar. 2025 were selected and followed up for 3 years. The recurrence and metastasis were counted. Patients with recurrence and metastasis were included in the poor prognosis group, and patients without recurrence and metastasis were included in the good prognosis group. The general data, preoperative serum sEC and COX-2 levels were compared between the two groups. The influencing factors of recurrence and metastasis of breast cancer after operation were analyzed. The dose-response relationship and predictive value of serum sEC and COX-2 levels with recurrence and metastasis were analyzed. Logistic regression equation (LR) model 1 was constructed according to conventional influencing factors, and LR model 2 was constructed according to conventional influencing factors combined with serum sEC and COX-2. The predictive efficacy and accuracy of the two models were analyzed by receiver operating characteristic curve (ROC) and calibration curve. According to the ratio of 7∶3, another 85 female breast cancer patients who underwent surgical treatment in Huai’an Hospital Affiliated to Xuzhou Medical University (Huai’an Second People’s Hospital) from May. 2023 to May. 2025 were selected for external validation (validation set) .Results:Among 198 breast cancer patients who underwent postoperative follow-up for 3 years, 3 cases were lost to follow-up, and the recurrence and metastasis rate among the 195 patients who completed the 3-year postoperative follow-up was 26.15% (51/195). In the good prognosis group, the proportion of TNM stage III was 20.14%, the proportion of lymph node metastasis was 11.81%, the preoperative serum carbohydrate antigen 153 (CA153) level was (54.19±10.84) U/mL, the carbohydrate antigen 125 (CA125) level was (46.03±10.27) U/mL, the sEC level was (1987.65±191.37) ng/mL, and the COX-2 level was (17.85±5.21) ng/mL. In the poor prognosis group, the proportion of TNM stage III was 45.10%, the proportion of lymph node metastasis was 35.29%, the preoperative serum CA153 level was (65.87±12.23) U/mL, the CA125 level was (57.76±11.51) U/mL, the sEC level was (2 295.37±261.48) ng/mL, and the COX-2 level was (10.42±3.16) ng/mL. Compared with the good prognosis group, they were increased ( t/χ 2=12.00, 14.11, 6.39, 6.79, 8.92, 12.00, P<0.05). Multivariate Logistic regression analysis showed that TNM stage III ( OR=2.078, 95%CI =1.569-2.751, P<0.05), lymph node metastasis ( OR=2.435, 95 %CI=1.843-3.216, P<0.05), serum CA153 ( OR= 1.180,95 %CI=1.026-1.357, P<0.05), CA125 ( OR=1.206,95 %CI=1.033-1.408, P<0.05), sEC ( OR=1.011,95 %CI= 1.007-1.015, P<0.05), COX-2 ( OR=1.378,95 %CI=1.128-1.683, P<0.05) were independent risk factors for postoperative recurrence and metastasis of breast cancer. Restricted cubic spline (RCS) analysis showed that serum sEC and COX-2 levels were positively correlated with the risk of recurrence and metastasis of breast cancer ( P<0.05). ROC analysis showed that serum sEC (AUC=0.762,95 %CI=0.696-0.820, P<0.05) and COX-2 (AUC=0.757,95 %CI=0.691-0.815, P<0.05) could be used as biological indicators to predict postoperative recurrence and metastasis of breast cancer. The AUC of LR model 1 in predicting postoperative recurrence and metastasis of breast cancer was 0.862 (95 %CI=0.805-0.907, P<0.05). The AUC of LR model 2 in predicting postoperative recurrence and metastasis of breast cancer was 0.965 (95 %CI= 0.891-0.983, P<0.05), which was significantly larger than that of LR model 1 ( Z=2.015, P<0.05). The calibration curve analysis showed that the calibration degree of LR model 1 in predicting postoperative recurrence and metastasis of breast cancer was good, and the prediction results were in good agreement with the actual observation results. The calibration degree of LR model 2 was high, and the prediction results were in good agreement with the actual observation results. The results of external validation showed that the sensitivity of LR model 2 in predicting postoperative recurrence and metastasis of breast cancer was 82.61%, the specificity was 93.55%, the accuracy was 90.59%, and the Kappa value was 0.762 (95 %CI: 0.549-0.974) . Conclusions:Preoperative serum sEC and COX-2 levels are independent risk factors for postoperative recurrence and metastasis of breast cancer. They can be used as biological indicators to predict postoperative recurrence and metastasis of breast cancer. Combined with conventional indicators, they can significantly improve the predictive efficacy.

Objective To conduct a unique and pioneering molecular epidemiological investigation of a case of Eurasian avian-like H1N1 swine influenza identified in Yunnan Province in 2022(the fourth such case in the province)and to understand its genetic characteristics so as to reveal its potential impact on human health.Methods Real-time fluorescent quantitative PCR detection technology was used for the nucleic acid testing of the case's pharyngeal swab samples,close contacts,and environmental samples from the living area.Positive samples were subjected to virus isolation using MDCK cells.Cell cultures were authenticated using erythrocyte agglutination assay with guinea pig blood and real-time fluorescence quantitative RT-PCR.Whole genome sequencing was performed using the Illumina MiseqNext-generation sequencing platform,and a phylogenetic tree was constructed using MEGA 7.0 software to analyze the genetic molecular characteristics.Results The first G5 genotype Eurasian avian-like H1N1 swine influenza virus in Yunnan Province was successfully isolated,and the whole genome sequence of the virus was obtained.This virus possessed the molecular characteristics associated with increased adaptability,virulence,or transmissibility in mammals and had a nucleotide consistency of 99.2％～99.7％with a porcine strain isolated in Jiangsu province.These findings underscored the potential threat this virus poses to human health.Conclusion The study underscores the importance of further monitoring swine influenza in preventing new influenza virus subtypes that can infect humans.