The present study aimed to explore whether hydrogen sulfide (H₂S) improved hypoxic pulmonary hypertension (HPH) in rats by inhibiting aerobic glycolysis-pyroptosis. Male Sprague-Dawley (SD) rats were randomly divided into normal group, normal+NaHS group, hypoxia group, and hypoxia+NaHS group, with 6 rats in each group. The control group rats were placed in a normoxic (21% O₂) environment and received daily intraperitoneal injections of an equal volume of normal saline. The normal+NaHS group rats were placed in a normoxic environment and intraperitoneally injected with 14 μmol/kg NaHS daily. The hypoxia group rats were placed in a hypoxia chamber, and the oxygen controller inside the chamber maintained the oxygen concentration at 9% to 10% by controlling the N₂ flow rate. An equal volume of normal saline was injected intraperitoneally every day. The hypoxia+NaHS group rats were also placed in an hypoxia chamber and intraperitoneally injected with 14 μmol/kg NaHS daily. After the completion of the four-week modeling, the mean pulmonary artery pressure (mPAP) of each group was measured using right heart catheterization technique, and the right ventricular hypertrophy index (RVHI) was weighed and calculated. HE staining was used to observe pathological changes in lung tissue, Masson staining was used to observe fibrosis of lung tissue, and Western blot was used to detect protein expression levels of hexokinase 2 (HK2), pyruvate dehydrogenase (PDH), pyruvate kinase isozyme type M2 (PKM2), nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), GSDMD-N-terminal domain (GSDMD-N), Caspase-1, interleukin-1β (IL-1β) and IL-18 in lung tissue. ELISA was used to detect contents of IL-1β and IL-18 in lung tissue. The results showed that, compared with the normal control group, there were no significant changes in all indexes in the normal+NaHS group, while the hypoxia group exhibited significantly increased mPAP and RVHI, thickened pulmonary vascular wall, narrowed lumen, increased collagen fibers, up-regulated expression levels of aerobic glycolysis-related proteins (HK2 and PKM2), up-regulated expression levels of pyroptosis-related proteins (NLRP3, GSDMD-N, Caspase-1, IL-1β, and IL-18), and increased contents of IL-1β and IL-18. These changes of the above indexes in the hypoxia group were significantly reversed by NaHS. These results suggest that H₂S can improve rat HPH by inhibiting aerobic glycolysis-pyroptosis.
Animals ; Rats, Sprague-Dawley ; Male ; Hypertension, Pulmonary/metabolism* ; Glycolysis/drug effects* ; Hydrogen Sulfide/therapeutic use* ; Hypoxia/complications* ; Rats ; Pyroptosis/drug effects*

The aim of this study was to investigate the contribution of lung zinc ions to pathogenesis of lung ischemia/reperfusion (I/R) injury in rats. Male Sprague Dawley (SD) rats were randomly divided into control group, lung I/R group (I/R group), lung I/R + low-dose zinc chloride group (LZnCl₂+I/R group), lung I/R + high-dose ZnCl₂ group (HZnCl₂+I/R group), lung I/R + medium-dose ZnCl₂ group (MZnCl₂+I/R group) and TPEN+MZnCl₂+I/R group (n = 8 in each group). Inductively coupled plasma mass spectrometry (ICP-MS) was used to measure the concentration of zinc ions in lung tissue. The degree of lung tissue injury was analyzed by observing HE staining, alveolar damage index, lung wet/dry weight ratio and lung tissue gross changes. TUNEL staining was used to detect cellular apoptosis in lung tissue. Western blot and RT-qPCR were used to determine the protein expression levels of caspase-3 and ZIP8, as well as the mRNA expression levels of zinc transporters (ZIP, ZNT) in lung tissue. The mitochondrial membrane potential (MMP) of lung tissue was detected by JC-1 MMP detection kit. The results showed that, compared with the control group, the lung tissue damage, lung wet/dry weight ratio and alveolar damage index were significantly increased in the I/R group. And in the lung tissue, the concentration of Zn²⁺ was markedly decreased, while the cleaved caspase-3/caspase-3 ratio and apoptotic levels were significantly increased. The expression levels of ZIP8 mRNA and protein were down-regulated significantly, while the mRNA expression of other zinc transporters remained unchanged. There was also a significant decrease in MMP. Compared with the I/R group, both MZnCl₂+I/R group and HZnCl₂+I/R group exhibited significantly reduced lung tissue injury, lung wet/dry weight ratio and alveolar damage index, increased Zn²⁺ concentration, decreased ratio of cleaved caspase-3/caspase-3 and apoptosis, and up-regulated expression levels of ZIP8 mRNA and protein. In addition, the MMP was significantly increased in the lung tissue. Zn²⁺ chelating agent TPEN reversed the above-mentioned protective effects of medium-dose ZnCl₂ on the lung tissue in the I/R group. The aforementioned results suggest that exogenous administration of ZnCl₂ can improve lung I/R injury in rats.
Animals ; Reperfusion Injury/pathology* ; Male ; Rats, Sprague-Dawley ; Rats ; Chlorides/administration & dosage* ; Lung/pathology* ; Zinc Compounds/administration & dosage* ; Apoptosis/drug effects* ; Caspase 3/metabolism* ; Cation Transport Proteins/metabolism*

Aconitum kusnezoffii is a perennial herbaceous medicinal plant of the family Ranunculaceae, with unique medicinal value. Damping off is one of the most important seedling diseases affecting A. kusnezoffii, occurring widely and often causing large-scale seedling death in the field. To clarify the species of the pathogen causing damping off in A. kusnezoffii and to formulate an effective control strategy, this study conducted pathogen identification, research on biological characteristics, and evaluation of fungicide inhibitory activity. Through morphological characteristics, cultural traits, and phylogenetic tree analysis, the pathogen causing damping off in A. kusnezoffii was identified as Rhizoctonia solani, belonging to the AG5 anastomosis group. The optimal temperature for mycelial growth of the pathogen was 25-30 ℃, with OA medium as the most suitable medium, pH 8 as the optimal pH, and sucrose and yeast as the best carbon and nitrogen sources, respectively. The effect of light on mycelial growth was not significant. In evaluating the inhibitory activity of 45 chemical fungicides, including 30% hymexazol, and 4 biogenic fungicides, including 0.3% eugenol, it was found that 30% thifluzamide and 50% fludioxonil had significantly better inhibitory effects on R. solani than other tested agents, with EC_(50) values of 0.129 6,0.220 6 μg·mL~(-1), respectively. Among the biogenic fungicides, 0.3% eugenol also showed an ideal inhibitory effect on the pathogen, with an EC_(50) of 1.668 9 μg·mL~(-1). To prevent the development of resistance in the pathogen and to reduce the use of chemical fungicides, it is recommended that the three fungicides above be used in rotation during production. These findings provide a theoretical basis for the accurate diagnosis and effective control strategy for R. solani causing damping off in A. kusnezoffii.
Fungicides, Industrial/pharmacology* ; Plant Diseases/microbiology* ; Rhizoctonia/growth & development* ; Aconitum/microbiology* ; Phylogeny ; Mycelium/growth & development*

In recent years, the study of single-cell transcriptome sequencing technology in the heterogeneity of astrocytes (astrocytes) after spinal cord injury (SCI) has provided new perspectives on post-traumatic nerve regeneration and repair. To provide a review on the research progress of single-cell sequencing technology in astrocytes after spinal cord injury (SCI), and to more comprehensively and deeply elaborate the application of single-cell sequencing technology in the field of astrocytes after SCI. Single-cell sequencing technology can analyse the transcriptomes of individual cells in a high-throughput manner, thus revealing fine differences in cell types and states. By using single-cell sequencing technology, the heterogeneity of astrocytes after SCI and their association with nerve regeneration and repair were revealed. In conclusion, the application of single-cell sequencing technology provides an important tool to reveal the heterogeneity of astrocytes after SCI, to further explore the mechanisms of astrocytes in SCI, and to develop intervention strategies targeting their regulatory mechanisms in order to improve the therapeutic efficacy of SCI. The discovery of changes in astrocyte transcriptome dynamics has improved researchers' understanding of spinal cord injury lesion progression and provided new insights into the treatment of spinal cord injury at different time points. To date, all of these findings need to be validated by more basic research and sufficient clinical trials. In the future, single-cell sequencing technology, through interdisciplinary collaboration with bioinformatics, computer science, tissue engineering, and clinical medicine, is expected to open a new window for the treatment of spinal cord injury.
Spinal Cord Injuries/metabolism* ; Astrocytes/cytology* ; Single-Cell Analysis/methods* ; Humans ; Animals ; Transcriptome ; Nerve Regeneration

Objective To develop a predictive model for postoperative mortality risk in patients with acute aortic dissection(AAD)using the Extreme Gradient Boosting(XGBoost)algorithm combined with Shapley Additive Explanation(SHAP),and to establish a prediction website to serve as a diagnostic and therapeutic support platform for clinicians and patients.Methods A retrospective cohort study design was adopted.Data from 782 AAD patients who underwent surgical treatment at the Fourth Hospital of Hebei Medical University from January 2013 to December 2023 were collected,including basic information and initial serum biomarker test results.Patients were randomly divided into training and test sets at a 7:3 ratio.An external validation set consisting of 313 AAD patients admitted to the Second Hospital of Hebei Medical University from January 2020 to December 2023 was also established for further model validation.Variables were screened using LASSO regression,and an XGBoost machine learning model was constructed and interpreted using SHAP.The predictive performance of the model was evaluated using receiver operating characteristic(ROC)curve analysis.Using the Shiny package,the XGBoost model was deployed to shinyapps.io to create a prediction website for postoperative mortality risk in AAD patients.One patient was selected by simple random sampling from the test set and the external validation set respectively for the prediction example on the Shiny webpage.Results The XGBoost model demonstrated high predictive performance for postoperative mortality in AAD patients,with area under the ROC curve(AUC)values of 0.928(95%CI 0.901-0.956)in the training set,0.919(95%CI 0.891-0.949)in the test set,and 0.941(95%CI 0.915-0.967)in the external validation set.SHAP values indicated the following order of variable importance in the model(from highest to lowest):"lactate dehydrogenase""blood chlorine""multiple organ injury""carbon dioxide combining power""prothrombin time""α-hydroxybutyric acid""creatine kinase isoenzyme""Stanford classification""combined use of bedside blood purification""gender""acute kidney injury""gastrointestinal bleeding""brain injury"and"shock".A risk prediction website for adverse postoperative outcomes in AAD patients was developed using XGBoost-SHAP method(https://dun-dunxiaolu.shinyapps.io/document/)and validated with examples.One randomly selected patient from each of the test and external validation sets was applied:the predicted mortality risk value for patient 1(who died postoperatively)was 0.9539,and that for patient 2(who survived postoperatively)was 0.0206.Conclusions The XGBoost-SHAP model demonstrates high accuracy in predicting postoperative mortality risk for AAD patients.The online prediction tool established based on this model enhances the identification efficiency of high-risk postoperative mortality patients.

AIM:This study aims to explore the role of asprosin(ASP)in regulating hepatic lipid anabolic metabolism,as well as the effects of ASP and its downstream pathways on hepatic lipid anabolic metabolism under obesity and exercise intervention.METHODS:To explore the effect of ASP on liver lipid synthesis under physiological condi-tion,5-week-old male C57BL/6J were randomly divided into 4 groups:wild-type(WT)group,WT+ASP group,knockout(KO)group,and KO+ASP group.The mice in KO and KO+ASP groups were ASP gene heterozygous KO mice,and the re-combinant ASP protein was intraperitoneally injected into the mice in WT+ASP and KO+ASP groups for one month before sampling.To explore the role of ASP on liver lipid synthesis under obesity and exercise intervention,5-week-old male C57BL/6J mice were randomly divided into 3 groups:normal control(NC)group,high-fat diet(HFD)group,and HFD+exercise group.After 10 weeks of HFD feeding,the mice in HFD group received no intervention,while those in HFD+ex-ercise group underwent 8 weeks of treadmill exercise intervention.The mice in both groups were fed with HFD during the intervention period.After the interventions,liver tissues were collected from the mice.Western blot and RT-qPCR meth-ods were used to detect the expression levels of ASP,fibroblast growth factor 21(FGF21),nuclear factor E2-related factor 2(NRF2),stearoyl-coenzyme A desaturase 1(SCD1)and fatty acid synthase(FASN)in the mouse liver.ELISA was used to detect the triglycerol(TG)and cyclic adenosine monophosphate(cAMP)levels in the mouse liver,and HE stain-ing and oil red O staining were performed to observe the morphological changes of mouse liver tissues.RESULTS:(1)Compared with WT mice,KO mice exhibited significantly reduced body weight and liver weight,while liver index and he-patic TG levels were significantly increased.The mRNA and protein levels of cAMP,ASP,FGF21 and NRF2 in the liver were significantly decreased,whereas the mRNA and protein levels of SCD1 and FASN were significantly increased.In the WT+ASP group,liver index and hepatic TG levels were significantly reduced,but there were no statistical differences in body weight and liver weight.The mRNA and protein levels of SCD1 and FASN in the liver were significantly de-creased,while the cAMP level and the mRNA and protein levels of NRF2 were significantly increased.The FGF21 mRNA level decreased,but the protein level increased.(2)Compared with KO mice,KO+ASP mice had significantly reduced hepatic TG level,with a certain degree of reduction in liver weight and liver index,but without statistical significance.The cAMP level and the mRNA and protein levels of ASP,FGF21 and NRF2 in the liver were significantly increased,while the mRNA and protein levels of SCD1 and FASN were significantly decreased.(3)Compared with NC mice,HFD mice showed increased body weight,liver weight,and hepatic TG level.The cAMP level and the mRNA and protein levels of NRF2 in the liver were significantly decreased,while the mRNA and protein levels of SCD1 and FASN were significant-ly increased.The FGF21 mRNA level increased,but its protein level decreased.The ASP mRNA level decreased,but its protein level increased.(4)Compared with HFD mice,HFD+exercise mice had significantly reduced body weight,liver weight,and hepatic TG level.The mRNA and protein levels of SCD1 and FASN in the liver were significantly decreased,while the cAMP level and the mRNA and protein levels of NRF2 were significantly increased.The FGF21 mRNA level de-creased,but its protein level increased.The ASP mRNA level increased,but its protein level decreased.CONCLU-SION:(1)The ASP exerts an inhibitory effect on hepatic lipogenesis through the cAMP/FGF21/NRF2 pathway.(2)Un-der HFD feeding condition,the protein expression of FGF21 decreases,leading to our speculation that ASP may develop resistance.Aerobic exercise intervention can attenuate ASP resistance under high-fat condition and alleviate hepatic lipid accumulation caused by HFD.

AIM:This study aims to explore the role of asprosin(ASP)in regulating hepatic lipid anabolic metabolism,as well as the effects of ASP and its downstream pathways on hepatic lipid anabolic metabolism under obesity and exercise intervention.METHODS:To explore the effect of ASP on liver lipid synthesis under physiological condi-tion,5-week-old male C57BL/6J were randomly divided into 4 groups:wild-type(WT)group,WT+ASP group,knockout(KO)group,and KO+ASP group.The mice in KO and KO+ASP groups were ASP gene heterozygous KO mice,and the re-combinant ASP protein was intraperitoneally injected into the mice in WT+ASP and KO+ASP groups for one month before sampling.To explore the role of ASP on liver lipid synthesis under obesity and exercise intervention,5-week-old male C57BL/6J mice were randomly divided into 3 groups:normal control(NC)group,high-fat diet(HFD)group,and HFD+exercise group.After 10 weeks of HFD feeding,the mice in HFD group received no intervention,while those in HFD+ex-ercise group underwent 8 weeks of treadmill exercise intervention.The mice in both groups were fed with HFD during the intervention period.After the interventions,liver tissues were collected from the mice.Western blot and RT-qPCR meth-ods were used to detect the expression levels of ASP,fibroblast growth factor 21(FGF21),nuclear factor E2-related factor 2(NRF2),stearoyl-coenzyme A desaturase 1(SCD1)and fatty acid synthase(FASN)in the mouse liver.ELISA was used to detect the triglycerol(TG)and cyclic adenosine monophosphate(cAMP)levels in the mouse liver,and HE stain-ing and oil red O staining were performed to observe the morphological changes of mouse liver tissues.RESULTS:(1)Compared with WT mice,KO mice exhibited significantly reduced body weight and liver weight,while liver index and he-patic TG levels were significantly increased.The mRNA and protein levels of cAMP,ASP,FGF21 and NRF2 in the liver were significantly decreased,whereas the mRNA and protein levels of SCD1 and FASN were significantly increased.In the WT+ASP group,liver index and hepatic TG levels were significantly reduced,but there were no statistical differences in body weight and liver weight.The mRNA and protein levels of SCD1 and FASN in the liver were significantly de-creased,while the cAMP level and the mRNA and protein levels of NRF2 were significantly increased.The FGF21 mRNA level decreased,but the protein level increased.(2)Compared with KO mice,KO+ASP mice had significantly reduced hepatic TG level,with a certain degree of reduction in liver weight and liver index,but without statistical significance.The cAMP level and the mRNA and protein levels of ASP,FGF21 and NRF2 in the liver were significantly increased,while the mRNA and protein levels of SCD1 and FASN were significantly decreased.(3)Compared with NC mice,HFD mice showed increased body weight,liver weight,and hepatic TG level.The cAMP level and the mRNA and protein levels of NRF2 in the liver were significantly decreased,while the mRNA and protein levels of SCD1 and FASN were significant-ly increased.The FGF21 mRNA level increased,but its protein level decreased.The ASP mRNA level decreased,but its protein level increased.(4)Compared with HFD mice,HFD+exercise mice had significantly reduced body weight,liver weight,and hepatic TG level.The mRNA and protein levels of SCD1 and FASN in the liver were significantly decreased,while the cAMP level and the mRNA and protein levels of NRF2 were significantly increased.The FGF21 mRNA level de-creased,but its protein level increased.The ASP mRNA level increased,but its protein level decreased.CONCLU-SION:(1)The ASP exerts an inhibitory effect on hepatic lipogenesis through the cAMP/FGF21/NRF2 pathway.(2)Un-der HFD feeding condition,the protein expression of FGF21 decreases,leading to our speculation that ASP may develop resistance.Aerobic exercise intervention can attenuate ASP resistance under high-fat condition and alleviate hepatic lipid accumulation caused by HFD.

Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannan-binding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP. Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays. Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05). Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.

The abuse and irrational use of antibiotics in human veterinary medicine has seriously endangered the ecological environment and human health.In this study,a fully automatic solid-phase disk extraction-stable isotope dilution-ultra-high performance liquid chromatography-mass spectrometry method for determination of 17 kinds of sulfonamides antibiotics(SAs)in water was established,which was then applied to determination of SAs in real samples including tap water,river water and seawater,respectively.Meanwhile,the residual characteristics were discussed and the ecological risks were assessed.With this method,1.0 L water sample with 0.5 g/L Na2EDTA(pH=3)was extracted and enriched by 3M SDB-XC disk,and eluted by 10 mL of mixture of methanol and acetone(1:1,V/V),and the pretreatment time of the sample was about 60 min per six samples.Under the optimized conditions,the linearity of the method for detection of 17 kinds of SAs ranged from 0.05 to 100 μg/L,with correlation coefficients(R2)>0.999.In addition,the detection limits(S/N=3)were as low as 0.012-0.052 ng/L,and the recoveries were in the range of 76%-110%,with relative standard deviations of 0.5%-9.6%(n=5).The results showed that no SAs was detected in tap water,while 3 and 9 kinds of SAs were detected in river water of Zhoushan,and seawater of Wenzhou Sea area in Zhejiang province,respectively.The total concentrations of the detected SAs were 0.875-21.826 ng/L and 1.024-20.768 ng/L in river water and seawater,respectively,and among which,sulfamethoxazole(SMX)was the predominant compound in river water and seawater,accounting for 81%and 74%of the total SAs,respectively.The ecological risk assessment showed that the risk quotients of the detected SAs in the river water and seawater in the study area for the three kinds of trophic organisms(algae,Daphnia and fish)were far less than 0.01,meaning that the ecological risk was low.

Purpose::Ischemia and hypoxia are the main factors limiting limb replantation and transplantation. Static cold storage (SCS), a common preservation method for tissues and organs, can only prolong limb ischemia time to 4 - 6 h. The normothermic machine perfusion (NMP) is a promising method for the preservation of tissues and organs, which can extend the preservation time in vitro by providing continuous oxygen and nutrients. This study aimed to evaluate the difference in the efficacy of the 2 limb preservation methods. Methods::The 6 forelimbs from beagle dogs were divided into 2 groups. In the SCS group ( n = 3), the limbs were preserved in a sterile refrigerator at 4 °C for 24 h, and in the NMP group ( n = 3), the perfusate prepared with autologous blood was used for the oxygenated machine perfusion at physiological temperature for 24 h, and the solution was changed every 6 h. The effects of limb storage were evaluated by weight gain, perfusate biochemical analysis, enzyme-linked immunosorbent assay, and histological analysis. All statistical analyses and graphs were performed using GraphPad Prism 9.0 one-way or two-way analysis of variance. The p value of less than 0.05 was considered to indicate statistical significance. Results::In the NMP group, the weight gained percentage was 11.72% ± 4.06%; the hypoxia-inducible factor-1α contents showed no significant changes; the shape of muscle fibers was normal; the gap between muscle fibers slightly increased, showing the intercellular distance of (30.19 ± 2.83) μm; and the vascular α-smooth muscle actin (α-SMA) contents were lower than those in the normal blood vessels. The creatine kinase level in the perfusate of the NMP group increased from the beginning of perfusion, decreased after each perfusate change, and remained stable at the end of perfusion showing a peak level of 4097.6 U/L. The lactate dehydrogenase level of the NMP group increased near the end of perfusion and reached the peak level of 374.4 U/L. In the SCS group, the percentage of weight gain was 0.18% ± 0.10%, and the contents of hypoxia-inducible factor-1α increased gradually and reached the maximum level of (164.85 ± 20.75) pg/mL at the end of the experiment. The muscle fibers lost their normal shape and the gap between muscle fibers increased, showing an intercellular distance of (41.66 ± 5.38) μm. The contents of vascular α-SMA were much lower in the SCS group as compared to normal blood vessels.Conclusions::NMP caused lesser muscle damage and contained more vascular α-SMA as compared to SCS. This study demonstrated that NMP of the amputated limb with perfusate solution based on autologous blood could maintain the physiological activities of the limb for at least 24 h.