Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO₂), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO₂ maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.

Intestinal barrier damage is a prominent feature of non-alcoholic fatty liver disease （NAFLD） and serves as a critical factor driving the progression from simple fatty liver to non-alcoholic steatohepatitis （NASH）， fibrosis， and cirrhosis. The "turbid pathogenic factors entering the blood" theory integrates classical traditional Chinese medicine （TCM） principles with contemporary disease evolution trends and research findings. It posits that endogenous turbid pathogenic factors within the body infiltrate the blood vessels， leading to impure and viscous blood quality， thereby triggering various diseases. Based on this theory， this article elucidated the pathogenic mechanism of NAFLD-associated intestinal barrier damage. It argued that in NAFLD， the liver loses its dredging function， and the spleen becomes obstructed and dysfunctional. Moreover， essential nutrients fail to be properly transformed， resulting in the internal generation of turbid pathogenic factors. This subsequently initiates a series of pathological changes， namely， "infiltration of phlegm-turbidity into the blood， eroding the intestinal mucosa"， "infiltration of glucose-turbidity into the blood， macerating and eroding the intestinal mucosa"， "infiltration of heat-turbidity into the blood， scorching and eroding the intestinal mucosa"， and "infiltration of stasis-turbidity into the blood， stagnating and eroding the intestinal mucosa"， ultimately causing intestinal barrier damage. Furthermore， guided by the "turbid pathogenic factors entering the blood" theory， this article explored TCM intervention strategies： employing medicinals targeting the liver meridian to address the root cause and reduce the generation and deposition of turbid pathogenic factors in the liver， administering blood-system medicinals to clear the blood and purge turbidity， thereby intercepting the progression of the disease mechanism， and applying tonifying medicinals to bolster healthy Qi and defend against turbid invasion， allowing the damaged intestinal mucosa to gradually heal. This article presented novel theoretical and medicinal perspectives for analyzing NAFLD-associated intestinal barrier damage based on the "turbid pathogenic factors entering the blood" theory， aiming to provide new entry points and broader horizons for related research and clinical practice.

Hepatocellular carcinoma （HCC）， the predominant subtype of primary liver cancer， ranks among the top in both incidence and mortality rates of malignant tumors in China. In its early stages， the disease may present with subtle or nonspecific symptoms， often leading to poor clinical prognosis and low patient survival rates， which makes it a significant public health concern. The pathogenesis is associated with multiple factors， including hepatitis virus infection， alcohol consumption， obesity， drug-induced liver injury， and immune disorders， which may interact synergistically to promote disease development. Currently， mainstream therapeutic approaches for HCC in modern medicine encompass surgical resection， liver transplantation， radiofrequency ablation， radiotherapy， and chemotherapy， but they all have certain limitations， such as large side effects and poor prognosis， imposing substantial psychological distress and financial strain on affected individuals. With a rich historical background in hepatic malignancy management， traditional Chinese medicine offers therapeutic benefits characterized by multi-targeted mechanisms， multi-level regulation， minimal adverse effects， and reduced likelihood of disease recurrence. It can not only enhance the curative effect， but also reduce the side effects of radiotherapy， chemotherapy， and surgery. Thus， it has attracted widespread attention. Extensive research has demonstrated that traditional Chinese medicine exhibits significant antitumor properties， along with notable anti-inflammatory and oxidative stress-reducing capabilities， and its mechanism may be related to the regulation of nuclear factor-kappa B （NF-κB） signaling pathway， which can affect multiple stages of hepatocarcinogenesis， such as cell proliferation， invasion， metastasis， and apoptosis. The mechanism of NF-κB signaling pathway in traditional Chinese medicine for HCC treatment has emerged as one of the pivotal research directions in current oncology studies. Based on the existing research foundation， a systematic literature review method was adopted to retrieve and analyze relevant Chinese and English literature in recent years. Integrating the molecular regulatory mechanisms of the NF-κB signaling pathway and its pivotal role in HCC pathogenesis and progression helped further explore the latest research advances in traditional Chinese medicine interventions targeting this pathway for HCC treatment. This approach may provide novel theoretical foundations and translational strategies for the prevention and management of HCC using traditional Chinese medicine.

OBJECTIVE To investigate the mechanism of cordycepin in delaying the transformation from acute kidney injury （AKI） to chronic kidney disease （CKD） （short for “AKI-CKD”）. METHODS Network pharmacology and bioinformatics were used to analyze and predict the signaling pathways of cordycepin in delaying AKI-CKD progression and its relationship with the ferroptosis pathway. Cell experiments were performed to verify the predicted results. Human renal tubular epithelial HK-2 cells were divided into blank group， cordycepin group， model group， and H 2 O 2 +cordycepin group. Except for the blank group and cordycepin group， all other groups were treated with 150 μmol/L H 2 O 2 for 72 h to induce persistent oxidative stress injury in cells. After 72 h of cordycepin （40 μmol/L） intervention， the protein and mRNA expression levels of glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4），nuclear factor-erythroid 2-related factor 2（Nrf2）， and heme oxygenase-1 （HO-1） in cells were detected. Furthermore， the Nrf2 inhibitor ML385 was added on the basis of H 2 O 2 +cordycepin to verify the role of the Nrf2/HO-1 pathway. RESULTS Network pharmacology and bioinformatics analysis showed that there were 42 overlapping genes related to AKI-CKD and ferroptosis that interact with cordycepin ferroptosis， among which 38 were annotated as ferroptosis-related genes. HO-1 and Nrf2 might be important targets for cordycepin to inhibit ferroptosis. The binding energies of cordycepin with Nrf2 and HO-1 proteins were －8.5 and －6.7 kcal/mol， respectively. Cell experiments showed that compared with the model group， the protein and mRNA expression levels of GPX4， Nrf2 and HO-1 were significantly increased （ P ＜0.05），while the protein and mRNA expression levels of ACSL4 were significantly decrease d （ P ＜0.05） in the H 2 O 2 +cordycepin group. After the addition of the Nrf2 inhibitor ML385， the effects of cordycepin on the above proteins and mRNA were significantly reversed （ P ＜0.05）. CONCLUSIONS Cordycepin can inhibit ferroptosis by activating the Nrf2/HO-1 pathway， reduce persistent oxidative stress injury of renal tubular epithelial cells， and delay the progression of AKI-CKD.

Objective To systematically evaluate the efficacy and safety of robot-assisted laparoscopic surgery (RAS) and conventional laparoscopic surgery (CLS) in hiatal hernia repair. Methods PubMed, Cochrane Library, CNKI, Web of Science, VIP, and Wanfang databases were searched to collect literature comparing the efficacy and safety of RAS and CLS for hiatal hernia repair published from their inception to November 7, 2023. The Newcastle-Ottawa Scale (NOS) was used to evaluate the quality of the included studies, and RevMan 5.4.1 software was used for meta-analysis. Results A total of 15 retrospective cohort studies with 18239 patients were finally included. The NOS scores of the included literature were all ≥7 points. Meta-analysis results showed that RAS was superior to CLS in terms of postoperative complications as the primary outcome [OR=0.56, 95%CI (0.42, 0.77), P<0.01]. There was no statistical difference between the two methods in terms of average operation time [MD=−0.74, 95%CI (−12.99, 11.51), P=0.91], average intraoperative blood loss [MD=−24.47, 95%CI (−54.80, 5.87), P=0.11], intraoperative complications [OR=0.76, 95%CI (0.29, 2.01), P=0.58], average postoperative hospital stay [MD=−0.24, 95%CI (−0.75, 0.27), P=0.36], postoperative GERD score [MD=−0.04, 95%CI (−0.41, 0.33), P=0.81], and 30-day readmission rate [OR=0.60, 95%CI (0.30, 1.20), P=0.15]. The cost of CLS surgery was less than that of RAS [SMD=1.59, 95%CI (1.16, 2.01), P<0.01]. Conclusion RAS has comparable efficacy and safety to CLS in hiatus hernia repair.

Objective:Explore the clinical diagnosis and treatment strategies of recurrent multiple chondritis in children with airway involvement as the main cause. Methods:From June 2021 to May 2023, five children with relapsing polychondritis were admitted to our department, all of whom met the Damiani criteria. Tracheotomy was performed in all five patients. Endoscopy indicated subglottic stenosis, with one case classified as grade Ⅲ and four cases as grade Ⅳ according to the Myer-Cotton classification. Enhanced CT scans of the head and neck showed fixed lumen narrowing and stenosis, including thickening of the soft tissue in the laryngeal cavity and partial absorption of the laryngotracheal cartilage anterior wall. In one patient, tracheal wall thickening and deformation with stenosis and calcification involved the posterior tracheal wall. One patient underwent "laryngeal tracheal reconstruction with hyoid graft, T-tube implantation, and bioabsorbable corticosteroid-eluting stent implantation", while three patients underwent "balloon dilatation, T-tube implantation, and bioabsorbable corticosteroid-eluting stent implantation" . Follow-up was performed every two months after the initial surgery, and three bioabsorbable corticosteroid-eluting stents were placed on the upper left, upper right, and lower T-tube, respectively, during this time. The entire follow-up period was six months. Results:All the 5 cases were in the outpatient department, of which 1 case had been extubation, 3 cases were still in the outpatient follow-up, and the remaining 1 case was still being treated in the rheumatology and immunology department due to poor control of the primary disease. Conclusion:In relapsing polychondritis children with airway involvement, tracheotomy could be used to quickly improve the symptoms of dyspnea in the disease progression. In the stable stage of the disease, the surgical method of 'Balloon dilatation + T-tube implantation + Bioabsorbable corticosteroid-eluting stents implantation' was adopted to reduce the secondary injury caused by surgical trauma as much as possible and improve the survival and quality of life of the children.
Humans ; Polychondritis, Relapsing/surgery* ; Child ; Male ; Female ; Tracheotomy ; Laryngostenosis ; Child, Preschool ; Trachea/surgery* ; Adolescent ; Stents

OBJECTIVE: To prepare decellularized nerve grafts from alpha-1, 3-galactosyltransferase (GGTA1) gene-edited pigs and explore their biocompatibility for xenotransplantation. METHODS: The sciatic nerves from wild-type pigs and GGTA1 gene-edited pigs were obtained and underwent decellularization. The alpha-galactosidase (α-gal) content in the sciatic nerves of GGTA1 gene-edited pigs was detected by using IB4 fluorescence staining and ELISA method to verify the knockout status of the GGTA1 gene, and using human sciatic nerve as a control. HE staining and scanning electron microscopy observation were used to observe the structure of the nerve samples. Immunofluorescence staining and DNA content determination were used to evaluate the degree of decellularization of the nerve samples. Fourteen nude mice were taken, and subcutaneous capsules were prepared on both sides of the spine. Decellularized nerve samples of wild-type pigs ( n=7) and GGTA1 gene-edited pigs ( n=7) were randomly implanted in the subcutaneous capsules. Blood was drawn at 1, 3, 5, and 7 days after implantation to detect neutrophil counting. RESULTS: IB4 fluorescence staining and ELISA detection showed that GGTA1 gene was successfully knocked out in the nerves of GGTA1 gene-edited pigs. HE staining showed that the structure of the decellularized nerve from GGTA1 gene-edited pigs was well preserved; the nerve basement membrane tube structure was visible under scanning electron microscopy; no cell nuclei was observed, and the extracellular matrix components was retained in the nerve grafts by immunofluorescence staining; and the DNA content was significantly reduced when compared with the normal nerves ( P<0.05). In vivo experiments showed that the number of neutrophils in the two groups were similar at 1, 3, and 7 days after implantation, with no significant difference ( P>0.05); only at 5 days, the number of neutrophils was significantly lower in the GGTA1 gene-edited pigs than in the wild-type pigs ( P<0.05). CONCLUSION The decellularized nerve grafts from GGTA1 gene-edited pigs have well-preserved nerve structure, complete decellularization, retain the natural nerve basement membrane tube structure and components, and low immune response after xenotransplantation through in vitro experiments.
Animals ; Transplantation, Heterologous ; Galactosyltransferases/genetics* ; Sciatic Nerve/immunology* ; Swine ; Tissue Engineering/methods* ; Humans ; Graft Rejection/prevention & control* ; Gene Editing ; Mice ; Mice, Nude ; Heterografts/immunology* ; Animals, Genetically Modified ; Tissue Scaffolds ; Decellularized Extracellular Matrix

BACKGROUND:Ilizarov bone transport is very effective in the treatment of open large tibial bone defects,but there are still complications,among which the difficulty of junction healing is one of the difficult points in treatment. OBJECTIVE:To investigate the effect of Ilizarov bone transport combined with antibiotic bone cement on junction healing after operation of open large tibial bone defect. METHODS:Totally 51 patients with open large tibial bone defect(bone defect>4 cm)admitted to Binzhou Medical University Hospital from August 2010 to January 2022 were selected,of which 28 received Ilizarov bone transport alone(control group)and 23 received Ilizarov bone transport combined with antibiotic bone cement treatment(trial group).External fixation time,bone healing time,bone healing index,visual analog scale score during bone removal,bone defect limb function,junction healing and complications at the final follow-up were statistically compared between the two groups. RESULTS AND CONCLUSION:(1)All the 51 patients were followed up for a mean of(22.53±5.77)months.External fixation time,bone healing time,bone healing index,postoperative infection rate,and non-healing rate of junction were less in the trial group than those in the control group(P<0.05).There was no significant difference between the two groups in visual analog scale scores at 6 months after the second surgery and in the functional excellence and good rate of limb with bone defect at the final follow-up(P>0.05).(2)These findings indicate that compared with the Ilizarov bone transport alone,Ilizarov bone transport combined with antibiotic bone cement treatment can promote the healing of open tibial fracture junction and increase the rate of bone healing.

Objective:To analyze the effect and safety of vibrating mesh atomization inhalation of pulmonary surfactant (PS) combined with nasal continuous positive airway pressure (NCPAP) in the treatment of neonatal respiratory distress syndrome (NRDS).Methods:150 Children with NRDS in department of pediatrics of the Second Affiliated Hospital of Air Force Medical University from May 2021 to May 2024 were enrolled as study subjects for a prospective cohort study. According to the propensity score matching, children who adopted traditional intratracheal infusion of PS combined with NCPAP were selected as infusion group (75 cases), and children who received vibrating mesh atomization inhalation of PS combined with NCPAP treatment were included in atomization group (75 cases). The therapeutic effect, breathing conditions, blood gas changes [pH value, arterial partial pressure of oxygen (PaO 2), arterial partial pressure of carbon dioxide (PaCO 2)] and NCPAP parameters changes [fraction of inspired oxygen (FiO 2), oxygenation index (OI), positive end-expiratory pressure (PEEP)] before and after treatment and incidence rates of adverse reactions were compared between groups. Normally distributed quantitative data were expressed as xˉ± s, and t test was used for inter-group comparison. Counting data were expressed as case(%), and χ2 test was used for inter-group comparison. Comparison of repeated measurement indicators at different time points using repeated meausurement analysis of variance. Results:There was no statistical significance in the total effective rate of treatment between groups ( P>0.05). The NCPAP treatment time and oxygen therapy time were (4.69±0.61) d and (14.52±1.16) d in atomization group, significantly shorter than (5.08±0.80) d and (15.30±1.28) d in infusion group ( t=3.36, 3.91, P=0.001, <0.001). The success rate of one-time weaning in atomization group was 88.00% (66/75), significantly higher than 73.33% (55/75) in infusion group ( χ2=5.17, P=0.023). After 3 and 7 days of treatment, the pH [(3 d : 7.31±0.04, 7 d: 7.34±0.03) and (3 d: 7.35±0.03, 7 d: 7.38±0.02)], PaO 2 [(3 d: (58.55±6.51) mmHg (1 mmHg=0.133 kPa), 7 d: (68.19±7.58) mmHg ) and (3 d: (65.16±7.24) mmHg, 7 d: (75.57±8.40) mmHg)] in infusion group and atomization group increased obviously over time, and the pH and PaO 2 were significantly higher in atomization group than those in infusion group ( t=6.93, 8.68, 5.88, 5.65, all P<0.001). The PaCO 2 in infusion group and atomization group (3 d: (48.45±5.38 ) mmHg, 7 d: (43.64±4.85 ) mmHg) and (3 d: (45.41±5.02) mmHg, 7 d: (40.35±4.59) mmHg) decreased obviously over time at 3 and 7 days of treatment, and the PaCO 2 in atomization group was significantly lower than that in infusion group ( t=3.58, 4.27, both P<0.001). After 3 days of treatment, FiO 2 [(41.06±4.67)% and (38.52±4.21)%] and PEEP [(4.39±0.19) cmH 2O (1 cmH 2O=0.098 kPa) and (4.28±0.13) cmH 2O] in infusion group and atomization group decreased obviously, and the indicators in atomization group were significantly lower than those in infusion group ( t=3.50, 4.14, both P<0.001). After 3 days of treatment, oxygenation index (OI) in infusion group and atomization group [(278.35±30.48) mmHg and (296.67±32.96) mmHg] increased obviously, and the OI in atomization group was significantly higher than that in infusion group ( t=3.53, P<0.001). The incidence of adverse reactions in atomization group was 4.00% (3/75), significantly lower than 13.34% (10/75) in infusion group ( χ2=4.13, P=0.042). Conclusion:Both vibrating mesh atomization inhalation of PS or traditional infusion of PS combined with NCPAP have similar efficacy in the treatment of NRDS, but vibrating mesh atomization inhalation of PS combined with NCPAP can more effectively improve the breathing conditions, blood gas indicators and NCPAP parameters of children, and has higher safety.

Objective To explore and verify the active components of Dachengqi decoction in regulating autophagy and its mechanism of protecting the intestinal mucosal barrier through network pharmacology and animal experiments.Methods The chemical components and autophagy-related target points of Dachengqi decoction were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database Analysis Platform(TCMSP)and GeneCards databases.The intersection of the drug target points and disease target points was taken and analyzed.The Cytoscape 3.10.2 software's Network Analyzer tool was used to analyze the drug components and target points,and the core target points were screened out to construct a traditional Chinese medicine compound regulatory network.The drug active component target point-disease network model and protein-protein interaction(PPI)network were visualized.Then,30 C57BL/6J mice were randomly divided into the Dachengqi decoction group,the intestinal infection group,and the control group,with 10 mice in each group.The intestinal infection group was given 200 μL/d of Klebsiella pneumoniae strain by gavage for 5 consecutive days,with a colony count of 109 CFU/mL,to create an intestinal infection model.The control group was given 200 μL/d of sterile normal saline by gavage.The Dachengqi decoction group(drug composition:Rhubarb 12 g,Aurantii Fructus 12 g,Magnolia Officinalis 24 g,Mirabilite 9 g,the drugs were dissolved in boiling distilled water to make a 1 kg/L solution)was given by gavage at a dose of 8 g·kg-1·d-1 for 3 consecutive days,and then given Klebsiella pneumoniae by gavage for 5 consecutive days on the 4th day.Detection indicators and methods:after the experiment,the mice were sacrificed and the terminal ileum tissues were collected.The tissues were stained with hematoxylin-eosin(HE),and the pathological changes of the intestinal mucosa were observed under a light microscope;immunofluorescence staining was used to observe the positive expressions of junction proteins ZO-1,Claudin-2,light chain 3-Ⅱ(LC3-Ⅱ),and Beclin-1 and the intestinal mucosal autophagy;the mRNA expression levels of autophagy genes were determined by polymerase chain reaction(PCR).Results The intersection of the obtained drug targets and disease targets yielded 111 potential autophagy-related targets for drug treatment of diseases.Key targets included β2-adrenergic receptor(ADRB2),heme oxygenase-1(HO-1),etc.,and the signaling pathways involved included AMP-activated protein kinase(AMPK)pathway,mammalian target of rapamycin(mTOR)pathway,etc.Animal experiments confirmed that the intestinal mucosal barrier function in the Dachengqi decoction group was better than that in the intestinal infection group,and the positive expression of microtubule-associated protein 1 lingt chain 3-Ⅱ(LC3-Ⅱ)and autophagy gene Beclin1 was significantly higher than that in the intestinal infection group.Transcriptome sequencing results showed that the key genes associated with autophagy and oxidative stress included ADRB2,HO-1,etc.The mRNA expression levels of ADRB2 and HO-1 in the Dachengqi decoction group were significantly higher than those in the intestinal infection group[HO-1 mRNA expression(FPKM):11.20±0.80 vs.6.63±0.53,ADRB2 mRNA expression(FPKM):6.98±0.54 vs.3.98±0.32,both P<0.01],verifying some of the predictions from network pharmacology.Conclusions Dachengqi decoction regulates autophagy through multiple components,multiple targets and multiple pathways,protecting the intestinal mucosal barrier function and reducing the translocation of intestinal microbiota.This lays a certain foundation for further in-depth research on the mechanism of reducing intestinal bacterial translocation by Dachengqi decoction.