BACKGROUND:We have successfully established an animal model of small ear pig in southern Yunnan province with internal tension relieving technique combined with autologous Achilles tendon for anterior cruciate ligament reconstruction,and verified the stability and reliability of the model.However,whether internal tension relieving technique can promote the ligamentalization process of autologous Achilles tendon graft has not been studied. OBJECTIVE:To investigate the differences in the process of ligamentalization between conventional reconstruction and internal reduction reconstruction of the anterior cruciate ligament by gross view,histology and electron microscopy. METHODS:Thirty adult female small ear pigs in southern Yunnan province were selected.Anterior cruciate ligament reconstruction was performed on the left knee joint with the ipsilateral knee Achilles tendon(n=30 in the normal group),and anterior cruciate ligament reconstruction was performed on the right knee joint with the ipsilateral knee Achilles tendon combined with the internal relaxation and enhancement system(n=30 in the relaxation group).The autogenous right forelimb was used as the control group;the anterior cruciate ligament was exposed but not severed or surgically treated.At 12,24,and 48 weeks after surgery,10 animals were sacrificed,respectively.The left and right knee joint specimens were taken for gross morphological observation to evaluate the graft morphology.MAS score was used to evaluate the excellent and good rate of the ligament at each time point.Hematoxylin-eosin staining was used to evaluate the degree of ligament graft vascularization.Collagen fibers and nuclear morphology were observed,and nuclear morphology was scored.Ultrastructural remodeling was evaluated by scanning electron microscopy and transmission electron microscopy. RESULTS AND CONCLUSION:(1)The ligament healing shape of the relaxation group was better at various time points after surgery,and the excellent and good rate of MAS score was higher(P<0.05).Moreover,the relaxation group could obtain higher ligament vascularization score(P<0.05).(2)The arrangement of collagen bundles and fiber bundles in the two groups gradually tended to be orderly,and the transverse fiber connections between collagen gradually increased and thickened,suggesting that the strength and shape degree of the grafts were gradually improved,but the ligament remodeling in the relaxation group was always faster than that in the normal group at various time points after surgery.(3)The diameter,distribution density,and arrangement degree of collagen fibers in the relaxation group were better than those in the normal group at all time points,especially in the comparison of collagen fiber diameter between and within the relaxation group(P<0.05).

Objective To investigate the impact of continuous nebulization therapy after lung transplantation on pulmonary function and related complications in lung transplant recipients. Methods A retrospective analysis was conducted on the general data of 71 recipients who underwent allogeneic lung transplantation at the Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University, from June 2013 to December 2024. Recipients were divided into observation group (those who continued nebulization therapy for more than 3 months after discharge) and control group (those who discontinued nebulization therapy on their own). The main observation indicators were pulmonary function indicators at 6 months after surgery, including forced expiratory volume in the first second as a percentage of predicted value (FEV₁% pred), forced vital capacity as a percentage of predicted value (FVC% pred), ratio of forced expiratory volume in the first second to forced vital capacity as a percentage of predicted value (FEV₁/FVC% pred), forced expiratory flow at 25%, 50% and 75% of forced vital capacity as a percentage of predicted value, and the percentage of predicted value of corrected carbon monoxide diffusion capacity measured by single-breath method, as well as the ratio of corrected carbon monoxide diffusion capacity to alveolar volume as a percentage of predicted value. Additionally, the annual incidence of postoperative pulmonary infections, survival rate and the rate of no severe airway complications were analyzed. Results At 6 months after lung transplantation, the FEV₁% pred and FVC% pred of the observation group were better than those of the control group [FEV₁% pred was 76% (60%, 91%) vs. 67% (62%, 78%), FVC% pred was (75 ± 13)% vs. (69 ± 11)%, both P<0.05]. The observation group had a lower annual incidence of pulmonary infections compared to the control group (P = 0.023), with a risk of 0.485 times that of the control group. There were no statistically significant differences between the two groups in median survival time and the rate of no severe airway complications (both P>0.05). Conclusions Continuous nebulization therapy after lung transplantation may effectively improve pulmonary function, reduce the annual incidence of pulmonary infections, and play a positive role in the long-term maintenance of pulmonary function.

Cardiac arrest (CA) is a critical condition in the field of cardiovascular medicine. Despite successful resuscitation, patients continue to have a high mortality rate, largely due to post CA syndrome (PCAS). However, the injury and pathophysiological mechanisms underlying PCAS remain unclear. Experimental animal models are valuable tools for exploring the etiology, pathogenesis, and potential interventions for CA and PCAS. Current CA animal models include electrical induction of ventricular fibrillation (VF), myocardial infarction, high potassium, asphyxia, and hemorrhagic shock. Although these models do not fully replicate the complexity of clinical CA, the mechanistic insights they provide remain highly relevant, including post-CA brain injury (PCABI), post-CA myocardial dysfunction (PAMD), systemic ischaemia/reperfusion injury (IRI), and the persistent precipitating pathology. Summarizing the methods of establishing CA models, the challenges encountered in the modeling process, and the mechanisms of PCAS can provide a foundation for developing standardized CA modeling protocols.
Animals ; Disease Models, Animal ; Post-Cardiac Arrest Syndrome/physiopathology* ; Heart Arrest/physiopathology* ; Humans ; Ventricular Fibrillation/complications*

OBJECTIVES: To verify whether hesperetin (Hes) alleviates doxorubicin (DOX)-induced cardiotoxicity by reducing inflammation via regulating the AMPK/NLRP3 pathway. METHODS: C57/bl6 mice and H9c2 cells treated with DOX to mimic cardiotoxicity were randomly divided into Sham (or control) group, DOX group, DOX+Hes group, DOX+Hes+compound C (CC, an AMPK inhibitor) group. Cardiac function and myocardial pathologies of the mice were evaluated, and the changes in H9c2 cell morphology and viability were assessed. Lactate dehydrogenase (LDH) activity in mouse myocardial tissues and H9c2 cells was measured using ELISA, and H9c2 cell apoptosis was detected with TUNEL staining. In both H9c2 cells and the myocardial tissues of the mice, cellular expression levels of TNF-α, IL-6 and IL-1β mRNAs and cleaved caspase-3, Bcl2, Bax, IL-1β, IL-18, p-AMPK, AMPK, p-mTOR, mTOR, NLRP3, ASC and caspase-1 proteins were detected using RT-PCR and Western blotting. RESULTS: DOX treatment caused cell swelling, decreased cell viability and increased LDH activity in H9c2 cells, resulting also in significantly increased cell apoptosis and cleaved caspase-3 expression and decreased Bcl2/Bax ratio. The DOX-treated mice showed obvious myocardial fiber swelling and inflammatory infiltration, decreased cardiac function and significantly increased myocardial LDH activity. In H9c2 cells, DOX treatment significantly increased the mRNA expressions of TNF-α, IL-6 and IL-1β and protein expressions of IL-1β and IL-18, lowered the expressions of p-AMPK and p-mTOR, and increased the expressions of NLRP3, ASC and caspase-1. Hes treatment obviously reduced these toxic effects of DOX in H9c2 cells, but its protective effects were blocked by application of compound C. CONCLUSIONS Hes reduces DOX-induced cardiotoxicity by inhibiting inflammation via regulating the AMPK/NLRP3 pathway.
Animals ; Doxorubicin/toxicity* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Mice, Inbred C57BL ; Mice ; Signal Transduction/drug effects* ; Cardiotoxicity ; AMP-Activated Protein Kinases/metabolism* ; Apoptosis/drug effects* ; Cell Line ; Myocytes, Cardiac/drug effects* ; Rats

Bacterial therapy has attracted increasing attention due to its special mechanism and abundant applications. With the flourishing development of synthetic biology, therapeutic genes have been introduced into engineering bacteria to improve their antitumor efficacy. However, it is difficult to spatiotemporally control the expression of these therapeutic genes at the tumor site in vivo, thereby considerably limiting the application of engineered bacteria in tumor treatment. To resolve this problem, we constructed a temperature-responsive bacterial strain capable of triggering the expression of exogenous genes in a laser-controllable way. Noxa, a pro-apoptotic protein, is chosen to test the expression of exogenous protein and its anti-tumor effect in engineered bacteria upon laser irradiation. Firstly, Noxa was fused to the C-terminus of the bacterial outer membrane protein cytolysin A (ClyA), and then the recombinant gene fragment ClyA-Noxa was inserted into the temperature-sensitive plasmid pBV220 and the recombinant plasmid was transformed into non-pathogenic Escherichia coli MG1655. Thus, we constructed the engineering strain (TRB@Noxa) that could express Noxa on the bacterial surface. TRB@Noxa could target and colonize the tumor tissue without causing notable host toxicity. The bacterial infection triggered thrombosis in the tumor tissue, resulting in the darkness of tumor sites. In a xenograft mouse tumor model, our strategy demonstrated precise tumor targeting and strong tumor inhibition. In conclusion, we successfully constructed a new engineering bacterial strain TRB@Noxa. TRB@Noxa combined with photothermal therapy could arrest tumor growth in the absence of photosensitizers, which represents an appealing method for antitumor therapy in the future.
Escherichia coli/radiation effects* ; Animals ; Humans ; Lasers ; Mice ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Neoplasms/therapy* ; Genetic Engineering ; Cell Line, Tumor ; Escherichia coli Proteins/genetics*

Cancer-associated thromboembolism(CAT)is one of the most common complications and the second direct cause of mortality in patients with malignant tumors,which seriously affect patients'life quality and prognosis.In recent years,with the deepening of mechanism researches of cancer and thrombosis,treatment strategies of CAT were also improved.The current status and progresses of clinical treatment of CAT were reviewed in this article.

Objective To investigate the underlying mechanism of microglia ferroptosis in smoke inhalation-induced(SII)brain injury.Methods Twenty SPF-grade male C57BL/6 mice were randomly divided into the control group,the SII group,the ferrostatin-1 group(Fer-1,2.5 mmol/kg)and the deferoxamine group(DFO,200 mg/kg),with 5 mice in each group.Mice in the Fer-1 group and the DFO group were intraperitoneally injected with Fer-1 and DFO 1,3 and 5 day after smoke exposure,respectively.The pathological changes of brain tissue were examined by HE staining and Prussian blue staining on the 6th day after smoke exposure.RT-qPCR was used to detect levels of inflammatory factors,brain tissue damage markers and ferroptosis markers.The contents of iron in mouse brain tissue were determined by double pyridine colorimetric assay.The contents of malondialdehyde(MDA)and lipid peroxide(LPO)in mouse brain tissue were determined by thiobarbituric acid(TBA)colorimetric assay.Superoxide dismutase(SOD)activity in mouse brain tissue was measured by xanthine oxidase assay kit.The contents of glutathione(GSH)in mouse brain tissue were determined by direct method of dithiodinitrobenzoic acid(DTNB)assay.BV2 cells were cultured in complete DMEM medium and divided into the control group,the erastin group(10 μmol/L),the Fer-1 group(erastinc stimulation combined with 5 mmol/L Fer-1 treatment)and the DFO group(erastinc stimulation combined with 50 mmol/L DFO treatment).After 24 h,cell viability was detected by CCK-8 assay,cell apoptosis was detected by Annexin V-FITC/PI flow cytometry,intracellular reactive oxygen species(ROS)production was detected by DCFDA staining,and mitochondrial membrane potential was detected by MitoTracker Red CMXRos staining.Results Compared with the control group,enhanced iron deposition and inflammation in brain tissue,elevated mRNA expression of inflammatory markers and damage markers in brain tissue,up-regulated ACSL4 and NCOA4 mRNA levels,down-regulated GPX4 and SLC7A11 mRNA levels,decreased GSH and SOD contents,and increased LPO and MDA contents were observed in brain tissue of the SII group(P＜0.05).The mRNA expression level of 7 member of solute carrier family 11(SLC7A11)was decreased in mice of the SII group.The contents of GSH and SOD were decreased,and the contents of LPO and MDA were increased(P＜0.05).Compared with the SII group,all the above parameters were reversed in the Fer-1 group and the DFO group,and the damage of mouse brain tissue was alleviated(P＜0.05).In BV2 cell experiments,compared with the control group,decreased survival rate of BV2 cells and increased apoptosis rate were found in the erastin group(P＜0.05),and increased intracellular ROS level and decreased mitochondrial membrane potential were also observed in the erastin-stimulated BV2 cells.The above parameters were opposite to those of the erastin group in the Fer-1 group and the DFO group(P＜0.05),and the oxidative damage of BV2 cells was alleviated.Conclusion The ferroptosis inhibitors Fer-1 and DFO can inhibit microglia ferroptosis and alleviate smoke inhalation-induced brain injury.

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori(Hp)infection in mice.Methods L.paracasei TK1501 was cultured for 32 h at 37℃in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×107 CFU/mL.The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption,cation exchange chromatography and HPLC,and its stability and antibacterial activity were assessed.The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection,which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL.Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments,and gastric tissues of the mice were collected for HE staining.Results L.paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid,base,and organic solvents.In the in vitro experiment,the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus,Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract.In the mouse models,treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice.Conclusion L.paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.

Objective To prepare a postbiotic using soybean fermentation product of Lactobacillus paracasei TK1501 and evaluate its inhibitory effect against Helicobacter pylori(Hp)infection in mice.Methods L.paracasei TK1501 was cultured for 32 h at 37℃in an anaerobic condition for solid substrate fermentation with a solid to water ratio of 1:1.5 in the substrate and an inoculation density of 5×107 CFU/mL.The postbiotic was isolated and purified using macroporous resin XAD-16N adsorption,cation exchange chromatography and HPLC,and its stability and antibacterial activity were assessed.The inhibitory effect of this postbiotic against Hp infection was evaluated in a mouse model with gastric mucosal Hp infection,which were treated with the postbiotic via gavage for 4 weeks at the dose of 0.02 or 0.1 mL.Serum levels of TNF-α and IL-1β of the mice were analyzed after the treatments,and gastric tissues of the mice were collected for HE staining.Results L.paracasei TK1501 postbiotic could be easily degraded by protease and had good thermal stability and tolerance to exposures to acid,base,and organic solvents.In the in vitro experiment,the postbiotic showed strong inhibitory effects in bacterial cultures of Staphylococcus aureus,Hp and other common pathogenic bacteria without obviously affecting the resident bacteria in the digestive tract.In the mouse models,treatment with the postbiotic at the dose of 0.1 mL significantly alleviated Hp infection and lowered the serum levels of TNF-α and IL-1β of the mice.Conclusion L.paracasei TK1501 postbiotic has strong inhibitory effects on Hp and Staphylococcus aureus but not on normal intestinal flora in mice.

OBJECTIVE To analyze the chemical components that were the absorbed in blood and liver tissue of rats after intragastric administration of aqueous extract from Abrus cantoniensis， and to speculate its possible metabolic pathways， providing reference for basic analysis of pharmacological substance in A. cantoniensis. METHODS Male SD rats were randomly divided into A. cantoniensis group （0.63 g/kg， calculated by crude drug） and blank group； they were given relevant drug solution/ultrapure water intragastrically. After a single dose， plasma and liver samples of rats in each group were collected. UPLC-Q-TOF/MS technology was used to identify chemical components that were absorbed in the blood and liver tissue of rats. RESULTS Totally， 30 chemical constituents were identified from the water extracts of A. cantoniensis， including alkaloids， flavonoids， organic acids， iridoids （such as L-abrine， schaftoside， isoshaftoside）. Ten prototype components and nine metabolites （such as decarboxylation and sulfation metabolites of protocatechuic acid， reduced sulfated metabolites of p-hydroxybenzoic acid） were identified from plasma samples； six prototype components and five metabolites （such as sulfated metabolites of p-hydroxybenzoic acid， decarboxylation and sulfation metabolites of p-hydroxybenzoic acid） were identified from liver samples. The main metabolic pathways included hydroxylation， demethylation， methylation， sulfation， glucuronidation， etc. CONCLUSIONS Alkaloids， flavonoids and organic acids are the main components of the aqueous extract from A. cantoniensis that are absorbed into the blood and liver， their metabolism mainly involves hydroxylation，demethylation， and sulfation.