Objective To explore the construction of heart preservation model of empty beating donor based on extracorporeal membrane oxygenation (ECMO). Methods From January 2022 to August 2023, 20 Guangxi Bama miniature pigs weighing 25-30 kg were selected, half male and half female. Under general anesthesia and heparinization, a midline thoracotomy was performed. The pericardium was cut after freeing the anterior and posterior vena cavae, and a perfusion needle was inserted near the brachiocephalic artery in the ascending aorta, connected to a blood collection bag to collect 500-600 mL of blood. The anterior and posterior vena cavae were ligated, the aorta was blocked and perfused with HTK solution to stop the heart beating. The superior and inferior vena cavae were cut off, the right pulmonary vein was decompressed, the aorta and left and right pulmonary arteries and veins were cut off, and the whole heart was removed. An ECMO device was used to continuously perfuse a cardioprotective solution mainly composed of oxygenated warm blood, maintaining the isolated pig heart beating for 8 hours, monitoring (once/hour) ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, inflammatory factors, myocardial enzymes, myoglobin, and troponin levels. Myocardial tissue was taken for hematoxylin-eosin (HE) staining to observe myocardial cell damage and evaluate the quality of heart preservation. Results Among the 20 isolated beating pig hearts, 17 successfully resumed beating, 3 experienced ventricular fibrillation, resuscitated after intracardiac electrical defibrillation, and all 20 pig hearts successfully beat for 8 hours. There was no statistical difference in ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, and inflammatory factors at each time point (P>0.05). There were statistical increases in myocardial enzymes, myoglobin, and troponin levels (P<0.05). HE staining results suggested that there was no severe myocardial damage. Conclusion ECMO technology can be used for pig heart preservation with good results, and this study provides experimental evidence for improving heart preservation research in clinical heart transplantation.

Objective:To explore the clinical effect of treating complex talar neck fractures with double-column fixation using plates and cannulated screws.Methods:A retrospective analysis was performed on the data of 13 patients with complex talar neck fractures treated with double-column fixation using plates and cannulated screws at Tianjin Hospital, Tianjin University from June 2019 to November 2023. There were 9 males and 4 females, with an age of 42.1±17.7 years (range, 15-66 years). There were 5 cases on the left and 8 cases on the right. Four cases were caused by traffic accidents, 8 by falling from a height, and 1 by a heavy object injury. According to the Hawkins classification, there were 12 cases of type II and 1 case of type III talar neck fractures. All the fractures were comminuted, including 12 cases of talar neck combined with talar body, and 3 cases combined with subluxation of subtalar joint. The time from injury to surgery was 3.3±1.6 d (range, 1-6 d). All patients were treated with anteromedial combined anterolateral approach, plate and cannulated screw double-column fixation pattern. The healing time of fractures and the occurrence of complications were recorded. Postoperative anteroposterior and lateral X-ray images were taken to assess the quality of fracture reduction based on the presence or absence of step-offs and angulation after reduction of fractures at the neck or body of the talus. The functional outcome was evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale.Results:All 13 patients were followed up for 33.8±15.2 months (range, 12-53 months). All fractures healed, and the healing time was 17.2±2.8 weeks (range, 13-23 weeks). Postoperative X-ray evaluation showed that 10 cases had anatomical reduction and 3 cases had near-anatomical reduction. After operation, there was no loosening or breakage of implant, loss of fracture reduction, irritation of skin and soft tissue by internal fixation. The AOFAS score was 88.1±13.0 points (range, 48-100 points), with 9 excellent cases, 3 good cases, and 1 poor case. Superficial skin necrosis in one surgical incision healed after dressing exchange. At the 1-year follow-up after surgery, 1 case developed avascular necrosis of the talus without collapse. And at the last follow-up (postoperative 13 to 53 months), 5 cases developed post-traumatic arthritis.Conclusion:Plate and cannulated screw double-column fixation in the treatment of complex talar neck fractures can achieve satisfactory reduction and strong fixation effects, which is beneficial in reducing complications related to poor reduction.

OBJECTIVE To investigate the role and mechanisms of the soluble guanylate cyclase(sGC)stimulator sGC003 in improving high altitude pulmonary edema(HAPE)in mice.METHODS Mice were randomly assigned to a normal control group,model group,model+dexamethasone 4 mg·kg-1 group(before modeling,intragastric administration of saline was performed once daily for 6 d,followed by intragastric administration of dexamethasone 4 mg·kg-1 on days 7 and 8),model+riociguat 10 mg·kg-1 group(before modeling,intragastric administration once a day for 7 d),and model+sGC003 5 and 10 mg·kg-1 groups(before modeling,intragastric administration once a day for 7 d).All groups except the normal control group received intratracheal instillation of lipopolysaccharide at a dose of 4 mg·kg-11 h after drug administration on day 7,followed by placement in a hypoxic environment to establish the HAPE model.After 24 h of modeling,the expiratory time,end-inspiratory pause,enhanced pause,and breathing frequency were measured,Lung tissue morphology was examined using HE staining,and lung tissue edema was assessed by determining the wet to dry weight ratio(W/D).The level of interleukin-1β(IL-1β)was determined using immunofluorescence staining.The phosphorylation level of vasodilator-stimulated phosphoprotein(VASP)in lung tissue was analyzed by Western blotting.Additionally,levels of sGC,hypoxia inducible factor-1α(HIF-1α),cyclic guanosine monophosphate(cGMP),IL-6,and IL-1βin serum were quantified using ELISA.RESULTS Compared with the normal control group,the model group had obvious pulmonary edema,and the lung W/D,IL-1β levels,expiratory time,end-inspiratory pause,enhanced pause,as well as serum levels of IL-1β,HIF-1α and IL-6 were significantly increased.Concurrently,the frequency of breathing and serum levels of sGC and cGMP were significantly decreased.Compared with model group,the expiratory time,end-inspiratory pause,enhanced pause,lung W/D and IL-1β levels,and serum levels of IL-1β,HIF-1α and IL-6 were significantly decreased in the model+sGC003 10 mg·kg-1 group;while the frequency of breathing,serum sGC and cGMP levels,phosphorylation level of VASP in lung tissues were significantly increased.CONCLUSION sGC003 can improve lung function,suppress pulmonary inflammation,and mitigate pulmonary edema in HAPE mice by activating the sGC/cGMP pathway.

OBJECTIVE To investigate the mechanism through which Chaixian Huashen decoction(CXHSD)ameliorates lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice.METHODS Component analysis:the components of CXHSD extract were analyzed via ultra-high performance liquid chromatography-high resolution mass spectrometry(UPLC-Q-Exactive HFX).Network pharma-cology analysis was conducted to predict the potential active components and underlying therapeutic targets of CXHSD for ALI treatment.① Animal experiment:mice were randomly divided into the normal control group,model(LPS)group,model+dexamethasone(DEX)4 mg·kg-1 group,model+CXHSD 10 g·kg-1 group,and model+CXHSD 20 g·kg-1 group.Except for the normal control group,ALI was induced in all the mice by intratracheal instillation of LPS.Model+CXHSD groups received daily intra-gastric administration of corresponding treatments for 7 consecutive days.The model+DEX group was administered saline intragastrically for the initial 5 d,followed by DEX for the next 2 d.ALI was induced by intratracheal instillation of LPS 5 mg·kg-1 1 h after the 6th administration of CXHSD/DEX.24 h after modeling,the severity of pulmonary edema was assessed using the wet to dry weight(W/D)ratio,and hematoxylin-eosin(HE)staining was used to evaluate histopathological damage.The levels of myeloperoxidase(MPO),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1β in lung tissue homogenates and serum were measured by enzyme-linked immunosorbent assay(ELISA).The total protein concentration in bronchoalveolar lavage fluid(BALF)was measured by bicinchoninic acid(BCA)assay.Immunohistochemistry and Western blotting were used to assess the expression levels of toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),zonula occludens-1(ZO-1)and occludin,as well as the phosphorylation level of nuclear factor-kappa B p65(NF-κB p65).② Cell experiment:RAW264.7 cells were divided into the cell control group,LPS 1 mg·L-1 group,LPS 1 mg·L-1+DEX 1 mg·L-1 group,and LPS 1 mg·L-1+CXHSD 50,100 and 200 mg·L-1 groups.After 24 h of culture,the nitric oxide(NO)content was measured with the nitrate reductase method,the levels of TNF-α,IL-1 β and IL-6 in the cell supernatants of each group were detected by ELISA.RESULTS Network pharmacology analysis indicated that CXHSD might alleviate ALI through the NF-κB pathway.① Com-pared with the normal control group,the W/D ratio was elevated,pathological injuries aggravated(such as alveolar wall thickening,inflammatory infiltration,and alveolar congestion),histopathological damage pronounced,MPO activity increased,and total protein concentrations in BALF raised in the model group,in which levels of TNF-α,IL-6 and IL-1 β in both lung tissue and serum became higher.Concur-rently,LPS increased the expressions of p-NF-κB p65,TLR4 and MyD88,but reduced the expressions of ZO-1 and occludin.Compared with the model group,model+CXHSD groups had their pulmonary edema and lung pathological injury ameliorated as evidenced by alleviated alveolar wall thickening,inflammatory infiltration and alveolar congestion.The levels of MPO,TNF-α,IL-1 β and IL-6 in both lung tissue and serum,and the total protein concentrations in BALF were significantly decreased in the model+CXHSD groups.Additionally,the expressions of TLR4,MyD88,and p-NF-κB p65 were significantly downregulated,while those of ZO-1 and occludin were prominently upregulated.② Compared with the cell control,the levels of TNF-α,IL-1 β,IL-6 and NO in the supernatant of RAW264.7 cells were signifi-cantly increased in the LPS group.Compared with the LPS group,in the supernatant of RAW264.7 cells treated with LPS+CXHSD at 100 mg·L-1,there was no significant difference in TNF-α levels.However,in the other groups treated with LPS+CXHSD,the levels of TNF-α,IL-1 β,IL-6,and the content of NO were significantly reduced.CONCLUSION CXHSD can alleviate LPS-induced ALI by inhibiting the TLR4/NF-κB pathway,attenuating inflammation,and preserving pulmonary barrier integrity.

Objective:To explore the clinical effect of treating complex talar neck fractures with double-column fixation using plates and cannulated screws.Methods:A retrospective analysis was performed on the data of 13 patients with complex talar neck fractures treated with double-column fixation using plates and cannulated screws at Tianjin Hospital, Tianjin University from June 2019 to November 2023. There were 9 males and 4 females, with an age of 42.1±17.7 years (range, 15-66 years). There were 5 cases on the left and 8 cases on the right. Four cases were caused by traffic accidents, 8 by falling from a height, and 1 by a heavy object injury. According to the Hawkins classification, there were 12 cases of type II and 1 case of type III talar neck fractures. All the fractures were comminuted, including 12 cases of talar neck combined with talar body, and 3 cases combined with subluxation of subtalar joint. The time from injury to surgery was 3.3±1.6 d (range, 1-6 d). All patients were treated with anteromedial combined anterolateral approach, plate and cannulated screw double-column fixation pattern. The healing time of fractures and the occurrence of complications were recorded. Postoperative anteroposterior and lateral X-ray images were taken to assess the quality of fracture reduction based on the presence or absence of step-offs and angulation after reduction of fractures at the neck or body of the talus. The functional outcome was evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale.Results:All 13 patients were followed up for 33.8±15.2 months (range, 12-53 months). All fractures healed, and the healing time was 17.2±2.8 weeks (range, 13-23 weeks). Postoperative X-ray evaluation showed that 10 cases had anatomical reduction and 3 cases had near-anatomical reduction. After operation, there was no loosening or breakage of implant, loss of fracture reduction, irritation of skin and soft tissue by internal fixation. The AOFAS score was 88.1±13.0 points (range, 48-100 points), with 9 excellent cases, 3 good cases, and 1 poor case. Superficial skin necrosis in one surgical incision healed after dressing exchange. At the 1-year follow-up after surgery, 1 case developed avascular necrosis of the talus without collapse. And at the last follow-up (postoperative 13 to 53 months), 5 cases developed post-traumatic arthritis.Conclusion:Plate and cannulated screw double-column fixation in the treatment of complex talar neck fractures can achieve satisfactory reduction and strong fixation effects, which is beneficial in reducing complications related to poor reduction.

OBJECTIVE To investigate the mechanism through which Chaixian Huashen decoction(CXHSD)ameliorates lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice.METHODS Component analysis:the components of CXHSD extract were analyzed via ultra-high performance liquid chromatography-high resolution mass spectrometry(UPLC-Q-Exactive HFX).Network pharma-cology analysis was conducted to predict the potential active components and underlying therapeutic targets of CXHSD for ALI treatment.① Animal experiment:mice were randomly divided into the normal control group,model(LPS)group,model+dexamethasone(DEX)4 mg·kg-1 group,model+CXHSD 10 g·kg-1 group,and model+CXHSD 20 g·kg-1 group.Except for the normal control group,ALI was induced in all the mice by intratracheal instillation of LPS.Model+CXHSD groups received daily intra-gastric administration of corresponding treatments for 7 consecutive days.The model+DEX group was administered saline intragastrically for the initial 5 d,followed by DEX for the next 2 d.ALI was induced by intratracheal instillation of LPS 5 mg·kg-1 1 h after the 6th administration of CXHSD/DEX.24 h after modeling,the severity of pulmonary edema was assessed using the wet to dry weight(W/D)ratio,and hematoxylin-eosin(HE)staining was used to evaluate histopathological damage.The levels of myeloperoxidase(MPO),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1β in lung tissue homogenates and serum were measured by enzyme-linked immunosorbent assay(ELISA).The total protein concentration in bronchoalveolar lavage fluid(BALF)was measured by bicinchoninic acid(BCA)assay.Immunohistochemistry and Western blotting were used to assess the expression levels of toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),zonula occludens-1(ZO-1)and occludin,as well as the phosphorylation level of nuclear factor-kappa B p65(NF-κB p65).② Cell experiment:RAW264.7 cells were divided into the cell control group,LPS 1 mg·L-1 group,LPS 1 mg·L-1+DEX 1 mg·L-1 group,and LPS 1 mg·L-1+CXHSD 50,100 and 200 mg·L-1 groups.After 24 h of culture,the nitric oxide(NO)content was measured with the nitrate reductase method,the levels of TNF-α,IL-1 β and IL-6 in the cell supernatants of each group were detected by ELISA.RESULTS Network pharmacology analysis indicated that CXHSD might alleviate ALI through the NF-κB pathway.① Com-pared with the normal control group,the W/D ratio was elevated,pathological injuries aggravated(such as alveolar wall thickening,inflammatory infiltration,and alveolar congestion),histopathological damage pronounced,MPO activity increased,and total protein concentrations in BALF raised in the model group,in which levels of TNF-α,IL-6 and IL-1 β in both lung tissue and serum became higher.Concur-rently,LPS increased the expressions of p-NF-κB p65,TLR4 and MyD88,but reduced the expressions of ZO-1 and occludin.Compared with the model group,model+CXHSD groups had their pulmonary edema and lung pathological injury ameliorated as evidenced by alleviated alveolar wall thickening,inflammatory infiltration and alveolar congestion.The levels of MPO,TNF-α,IL-1 β and IL-6 in both lung tissue and serum,and the total protein concentrations in BALF were significantly decreased in the model+CXHSD groups.Additionally,the expressions of TLR4,MyD88,and p-NF-κB p65 were significantly downregulated,while those of ZO-1 and occludin were prominently upregulated.② Compared with the cell control,the levels of TNF-α,IL-1 β,IL-6 and NO in the supernatant of RAW264.7 cells were signifi-cantly increased in the LPS group.Compared with the LPS group,in the supernatant of RAW264.7 cells treated with LPS+CXHSD at 100 mg·L-1,there was no significant difference in TNF-α levels.However,in the other groups treated with LPS+CXHSD,the levels of TNF-α,IL-1 β,IL-6,and the content of NO were significantly reduced.CONCLUSION CXHSD can alleviate LPS-induced ALI by inhibiting the TLR4/NF-κB pathway,attenuating inflammation,and preserving pulmonary barrier integrity.

OBJECTIVE To investigate the role and mechanisms of the soluble guanylate cyclase(sGC)stimulator sGC003 in improving high altitude pulmonary edema(HAPE)in mice.METHODS Mice were randomly assigned to a normal control group,model group,model+dexamethasone 4 mg·kg-1 group(before modeling,intragastric administration of saline was performed once daily for 6 d,followed by intragastric administration of dexamethasone 4 mg·kg-1 on days 7 and 8),model+riociguat 10 mg·kg-1 group(before modeling,intragastric administration once a day for 7 d),and model+sGC003 5 and 10 mg·kg-1 groups(before modeling,intragastric administration once a day for 7 d).All groups except the normal control group received intratracheal instillation of lipopolysaccharide at a dose of 4 mg·kg-11 h after drug administration on day 7,followed by placement in a hypoxic environment to establish the HAPE model.After 24 h of modeling,the expiratory time,end-inspiratory pause,enhanced pause,and breathing frequency were measured,Lung tissue morphology was examined using HE staining,and lung tissue edema was assessed by determining the wet to dry weight ratio(W/D).The level of interleukin-1β(IL-1β)was determined using immunofluorescence staining.The phosphorylation level of vasodilator-stimulated phosphoprotein(VASP)in lung tissue was analyzed by Western blotting.Additionally,levels of sGC,hypoxia inducible factor-1α(HIF-1α),cyclic guanosine monophosphate(cGMP),IL-6,and IL-1βin serum were quantified using ELISA.RESULTS Compared with the normal control group,the model group had obvious pulmonary edema,and the lung W/D,IL-1β levels,expiratory time,end-inspiratory pause,enhanced pause,as well as serum levels of IL-1β,HIF-1α and IL-6 were significantly increased.Concurrently,the frequency of breathing and serum levels of sGC and cGMP were significantly decreased.Compared with model group,the expiratory time,end-inspiratory pause,enhanced pause,lung W/D and IL-1β levels,and serum levels of IL-1β,HIF-1α and IL-6 were significantly decreased in the model+sGC003 10 mg·kg-1 group;while the frequency of breathing,serum sGC and cGMP levels,phosphorylation level of VASP in lung tissues were significantly increased.CONCLUSION sGC003 can improve lung function,suppress pulmonary inflammation,and mitigate pulmonary edema in HAPE mice by activating the sGC/cGMP pathway.

Objective:To compare the early therapeutic effects of internal fixation with percutaneous minimally invasive hollow nail assisted by electromagnetic navigation robot and guided by C-arm in the treatment of Tile type C pelvic fracture.Methods:A retrospective cohort study was conducted to analyze the clinical data of 32 patients with Tile type C pelvic fracture admitted to Weihai Central Hospital from January 2020 to March 2022, including 18 males and 14 females; aged 36-60 years [(44.1±3.9)years]. Among them, 17 patients were treated with internal fixation with percutaneous minimally invasive hollow nail assisted by electromagnetic navigation robot (electromagnetic navigation group), and 15 with internal fixation with percutaneous minimally invasive hollow nail guided by C-arm (C-arm guidance group). Operative time, intraoperative blood loss, sacroiliac screw placement time, pubic branch screw placement time, ambulation time and fracture healing time were compared between the two groups. Visual analog scale (VAS), Majeed function score and complication rate at 1 day, 6 months, 12 months after surgery and at the last follow-up were compared between the two groups.Results:All the patients were followed up for 12-24 months [(15.4±0.5)months]. The operative time and intraoperative blood loss in the electromagnetic navigation group were (42.0±2.5)minutes and (10.9±2.6)ml, shorter or less than (50.0±3.5)minutes and (14.9±3.1)ml in the C-arm guidance group (all P<0.01). The placement time of sacroiliac screw and pubic branch screw in the electromagnetic navigation group was (12.4±0.2)minutes and (10.1±0.3)minutes, shorter than (15.3±0.3)minutes and (13.2±0.3)minutes in the C-arm guidance group (all P<0.01). The ambulation time was (3.2±0.4)weeks in the electromagnetic navigation group, earlier than (3.5±0.4)weeks in the C-arm guidance group ( P<0.05). There was no significant difference in fracture healing time between the two groups ( P>0.05). VAS scores of the electromagnetic navigation group were (4.4±0.3)points and (1.1±0.1)points at 1 day and 6 months after surgery respectively, lower than those of the C-arm guidance group [(4.8±0.4)points and (1.2±0.3)points] ( P<0.05 or 0.01). Majeed function scores of the electromagnetic navigation group were (37.3±1.1)points and (88.5±1.4)points at 1 day and 6 months after surgery respectively, higher than those of the C-arm guidance group [(30.7±4.2)points and (82.6±1.8)points] (all P<0.01). There were no significant differences in VAS and Majeed scores at 12 months after surgery and at the last follow-up between the two groups (all P>0.05). There was no significant difference in the incidence of postoperative complications between the two groups ( P>0.05). Conclusion:Compared with C-arm guidance, electromagnetic navigation robot-assisted internal fixation with percutaneous minimally invasive hollow nail for Tile type C pelvic fracture can reduce operative time and intraoperative blood loss, shorten screw placement time and ambulation time, relieve pain and improve functional recovery at early stage.

Objective To explore the relationship between non-high density lipoprotein cholesterol(non-HDL-C)level and leptomeningeal collateral circulation in patients with acute middle cerebral artery occlusion.Methods A total of 85 patients with first-onset acute cerebral infarction with middle cerebral artery M1 segment occlusion were enrolled.According to the results of DSA,LMC circulation was assessed by American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology Collateral Circulation Assess-ment System.All patients were assigned to better LMC circulation group(score 2～4,n = 30)and worse LMC circulation group(score 0～1,n = 55),and the levels of non-HDL-C were compared between the two groups.Results The levels of LDL-C and non-HDL-C in worse LMC circulation group were significantly higher than those of the better LMC circulation group(P = 0.026,P = 0.010).non-HDL-C was an independent risk factor for the worse LMC circulation(OR = 3.019,95%CI:1.053～8.658,P = 0.04).LMC circulatory score of patients was negatively correlated with the levels of non-HDL-C level(r =-0.228,P = 0.036).The AUC of non-HDL-C predicted for the worse LMC circulation was 0.638(95%CI:0.521～0.755,P = 0.036).Conclusions non-HDL-C in patients with acute cerebral infarction was significantly related to worse LMC circulation,and was a risk factor for worse LMC circulation.It is suggested that the higher expression of non-HDL-C could be used to predict worse LMC circulation as a serological indicator.

Plant fungal pathogens secrete numerous proteins into the apoplast at the plant-fungus contact sites to facilitate colonization.However,only a few secretory proteins were functionally characterized in Magnaporthe oryzae,the fungal pathogen causing rice blast disease worldwide.Asparagine-linked glycosylation 3(Alg3)is an a-l,3-mannosyltransferase functioning in the N-glycan synthesis of N-glycosylated secretory proteins.Fungal pathogenicity and cell wall integrity are impaired in Aalg3 mutants,but the secreted proteins affected in Aalg3 mutants are largely unknown.In this study,we compared the secretomes of the wild-type strain and the Aalg3 mutant and identified 51 proteins that require Alg3 for proper secretion.These proteins were predicted to be involved in metabolic processes,interspecies interactions,cell wall organization,and response to chemicals.Nine proteins were selected for further validation.We found that these proteins were localized at the apoplastic region surrounding the fungal infection hyphae.Moreover,the N-glycosylation of these proteins was significantly changed in the Aalg3 mutant,leading to the decreased protein secretion and abnormal protein localization.Furthermore,we tested the biological functions of two genes,INV1(encoding invertase 1,a secreted invertase)and AMCase(encoding acid mammalian chinitase,a secreted chitinase).The fungal virulence was significantly reduced,and the cell wall integrity was altered in the Ainv1 and Aamcase mutant strains.Moreover,the N-glycosylation was essential for the function and secretion of AMCase.Taken together,our study provides new insight into the role of N-glycosylated secretory proteins in fungal virulence and cell wall integrity.