Objective:To explore the distribution and clearance of 99mTc labeled diethylenetriamine pentaacetic acid(99mTc-DTPA)in different brain regions of adult rats after administration through brain extracellular space(ECS)pathway.Methods:After the injection of a volume of 2 μL and radioactive activity of about 3.7 MBq(100 μCi)of 99mTc-DTPA into the caudate nucleus and thalamus of SD rats through stereotactic positioning of rat brain,the single photon emission computed tomography/computed tomography(SPECT/CT)for small animals was used for imaging at different time points,and the dyna-mic distribution and clearance of the tracer in the whole body were observed continuously.The SD rats were injected with 99mTc-DTPA into thalamus and caudate nucleus respectively for biological distribution in vivo.They were put to death 4 h later.Their blood and urine were collected.The brain,cerebellum,heart,liver,spleen,lung,and kidney were taken and weighed by γ counter to measure its radioactivity.Results:SPECT/CT imaging results showed that after 99mTc-DTPA was administered through brain ECS,the radioactivity was concentrated in the brain,kidney and bladder.The tracer administered to the left caudate nucleus was preferentially drained to the right cerebellum,while the tracer administered to the right caudate nucleus was preferentially drained to the left cerebellum.There was a phenomenon of"con-tralateral cerebellar dominant drainage"in the caudate nucleus.The thalamic area preferentially drained to the ipsilateral cerebellum after administration.Four hours after administration via ECS,high radioac-tive uptake appeared in urine,cerebellum and brain,followed by blood and kidney.The radioactive up-take values of heart,liver,spleen and lung were low,which were mainly excreted through urinary sys-tem.Conclusion:Intracerebral ECS administration is a promising method of administration,but there are significant differences in distribution and clearance in different brain regions.This study further ex-pands the content and significance of"ECS regions",and also provides an important theoretical founda-tion for the treatment of encephalopathy and the research of new drugs through brain ECS in the future.

The study explored the therapeutic effect and mechanism of the compound composed of tangerine peel,plum,agrimony and hawthorn on Eimeria tenella in chicks.In the experiment,144 chicks were divided into six groups:the blank group,the model group,the diclazuril group,and the high-dose,medium-dose and low-dose traditional Chinese medicine compound groups.The 14-day-old chickens were infected,the body weight and mortality were recorded,and the samples were dis-sected at 21 days of age.The anticoccidial effect of Chenpi compound at different doses was evalua-ted by calculating anticoccidial index,feed conversion rate and detecting serum cytokine levels and related mRNA expression levels.The results showed that the anticoccidial index of the middle dose group was 172.86,and the anticoccidial index of the low dose group was 158.98.In the middle and low dose groups,the levels of IL-1β,IL-6 and MDA in serum decreased significantly,while the levels of IL-4,IL10,T-AOC,SOD,CAT and GSH-Px increased significantly,and the mRNA ex-pression levels of related inflammation and antioxidant pathways changed.Further studies found that the medium-dose compound can regulate the ChTLR15/ChMyD88/ChNF-κB/ChNLRP3/Caspase-1 inflammatory signaling pathway,activate the Nrf2/Keap1/HO-1 antioxidant signaling pathway,and enhance the body's anti-inflammatory and antioxidant capacity;at the same time,it can increase the expression of intestinal tight junction ZO-1 and Occludin,repair the damaged in-testinal barrier,and play an anticoccidial role.The results showed that the middle dose of tangerine peel compound had a good effect in the treatment of coccidiosis,and its mechanism involved the regulation of anti-inflammatory and antioxidant pathways.

Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.

Objective To investigate the predictive value of serum vasoactive intestinal peptide(VIP)and 5-hydroxytryptamine(5-HT)levels on the outcome of spinal cord electrical stimulation(SCS)in patients with postherpetic neuralgia(PHN).Methods A total of 96 PHN patients who received SCS treatment in Ziy-ang Central Hospital from January 2022 to December 2023 were selected.According to the disease outcomes of all PHN patients after 6 months of treatment,a good group(n=71)and a poor group(n=25)were set up.The clinical data of the two groups were collected and the serum VIP and 5-HT levels were detected in all pa-tients before treatment.The predictive value of serum VIP and 5-HT on disease outcome after SCS treatment in PHN patients was evaluated by receiver operating characteristic(ROC)curve,and the influencing factors of disease outcome after SCS treatment in PHN patients was explored by multivariate Logistic steppe gression a-nalysis.Results The levels of serum VIP and 5-HT in poor group were higher than those in good group(P＜0.05).The area under the curve(AUC)of serum VIP and 5-HT for predicting the disease outcome of PHN patients after SCS treatment were 0.829(95％CI:0.779-0.874)and 0.743(95％CI:0.693-0.793),respec-tively,and the AUC of combined prediction was 0.941(0.891-0.986).There were no significant differences in age,gender,body moss index,education,location of onset,hypertension and drinking history between the two groups(P＞0.05).The time of initial hospital admission in the poor group was longer than that in the good group,skin rash area in the poor group was larger than that in the good group,and diabetes mellitus and smoking history in the poor group were higher than those in the good group(P＜0.05).The time of admis-sion for initial treatment＞3 d(OR=2.188,95％CI:1.383-3.461),skin rash area＞10 cm2(OR=2.018,95％CI:1.283-3.173),diabetes mellitus(OR=2.264,95％CI:1.379-3.717),serum VIP level ≥41.78 ng/L(OR=3.022,95％CI:1.685-5.420),serum 5-HT level ≥99.27 ng/mL(OR=3.579,95％CI:1.885-6.793)were the influencing factors of disease outcome after SCS treatment in PHN patients(P＜0.05).Con-clusion The elevated levels of serum VIP and 5-HT before treatment are associated with poor outcomes after SCS in patients with PHN,and could be used as potential markers to predict the outcomes of SCS in patients with PHN.

Objective To explore the molecular mechanisms by which cytochrome P4502C regulates ferroptosis in hu-man retinal microvascular endothelial cells(hRMECs)under high-glucose conditions.Methods Human retinal microvas-cular endothelial cells(hRMECs)were cultured in vitro and divided into six groups:the NG group(cells cultured in ser-um-free DMEM medium containing 5.5 mmol·L-1 glucose);the NG+fenofibrate group(10 μmol·L-1 fenofibrate added to the NG group);the NG+rifampicin group(10 μmol·L-1 rifampicin added to the NG group);the HG group(cells cul-tured in serum-free DMEM medium containing 25.5 mmol·L-1 glucose);the HG+fenofibrate group(10 μmol·L-1 feno-fibrate added to the HG group);and the HG+rifampicin group(10 μmol·L-1 rifampicin added to the HG group).After 48 hours of culture,subsequent experiments were conducted.Cell viability was assessed using the CCK-8 assay.The levels of malondialdehyde(MDA)and iron content in hRMECs were measured using commercial assay kits.The relative fluores-cence intensity of Liperfluo was quantified by flow cytometry.The mRNA expression levels of glutathione peroxidase 4(GPX4),cytochrome P4502C,and acyl-CoA synthetase long-chain family member 4(ACSL4)in hRMECs were analyzed using real-time quantitative PCR.The protein expression levels of GPX4,cytochrome P4502C,and ACSL4 in hRMECs were evaluated using Western blot analysis.Results Optimal concentration analysis showed that the 10 μmnol·L-1 fenofibrate group had the highest cell viability compared to the HG group(P＜0.05),while the 10 μmol·L-1 rifampicin group had the lowest cell viability(P＜0.05).Thus,10 μmol·L-1 was selected as the optimal concentration for both fenofibrate and rif-ampicin for subsequent experiments.Cell viability in the NG,NG+fenofibrate,NG+rifampicin,HG,HG+fenofibrate,and HG+rifampicin groups was(100.00±7.85)％,(102.15±9.07)％,(69.97±4.22)％,(61.74±6.13)％,(83.64±7.58)％,and(56.96±5.34)％,respectively.Compared with the NG group,the HG group had significantly lower cell via-bility(P＜0.05).Compared with the HG group,the HG+fenofibrate group had significantly higher cell viability(P＜0.05).Compared with the NG group,the HG group had higher levels of MDA,iron content,and Liperfluo relative fluores-cence intensity(all P＜0.05).Compared with the HG group,the HG+fenofibrate group had lower levels of MDA,iron content,and Liperfluo relative fluorescence intensity(all P＜0.05);the HG+rifampicin group had higher levels of these parameters(all P＜0.05).Compared with the NG group,the HG group had higher mRNA and protein expression levels of cytochrome P4502C and ACSL4,but lower levels of GPX4 mRNA and protein(all P＜0.05).Compared with the HG group,the HG+fenofibrate group had lower mRNA and protein expression levels of cytochrome P4502C and ACSL4,but higher lev-els of GPX4 mRNA and protein(all P＜0.05);the HG+rifampicin group had higher mRNA and protein expression levels of cytochrome P4502C and ACSL4,but lower levels of GPX4 mRNA and protein(all P＜0.05).Conclusion Cytochrome P4502C can promote ferroptosis of hRMECs under high-glucose conditions by regulating the GPX4/ACSL4 signaling axis;in-hibition of cytochrome P4502C expression can exert a protective effect on DR;modulation of the expression levels or bal-ance of the key factors GPX4 and ACSL4 in this pathway may hold promise as a potential strategy to slow the progression of DR.

AIM:To establish a cell model of visceral hypersensitivity and nerve hyperplasia in irritable bowel syndrome(IBS)by conducting an in vitro co-culture of mouse P815 mast cells and N2a nerve cells and explore its possible mechanism.METHODS:Enzyme-linked immunosorbent assay(ELISA)with three replicates was used to confirm the C48/80-induced P815 degranulation.The length of neurites was observed under bright field microscopy to determine the number of differentiated neurons,thereby selecting the concentration of retinoic acid(RA)for stimulating the differentia-tion of N2a cells,with four replicates.A co-culture system of P815 and N2a cells was established using Transwell cham-bers with four replicates.The following groups were established:N2a cells cultured alone,N2a cells co-cultured with P815 cells,N2a cells co-cultured with P815 cells plus C48/80,and N2a cells plus RA group.After co-culturing,the num-ber of differentiated N2a cells was observed under bright field.The expression of nerve growth factor(NGF),tyrosine ki-nase receptor A(TRKA),growth-associated protein-43(GAP43),neuron-specific enolase(NSE),synapsin(SYN),and postsynaptic density protein-95(PSD-95)at both protein and gene levels in N2a cells was detected using Western blot and polymerase chain reaction(PCR),with four replicates.RESULTS:The best condition for N2a differentiation was stimulation with 10 μmol/L RA for 24 hours,whereas the best condition for degranulation was stimulation of P815 cells with 20 mg/L C48/80 for 24 hours.Compared with N2a cells cultured alone,the differentiation ratio of the N2a+P815+C48/80 and N2a+RA groups was significantly increased(P<0.01),and the protein and mRNA expressions of NGF,TRKA,GAP43,NSE,SYN,and PSD-95 were significantly increased(P<0.05).CONCLUSION:Our results revealed that mast cell degranulation enhances the level of nerve hyperplasia in enteric nerve cells and promotes changes in nerve structure and function.Synaptic remodeling regulated by abnormal expression of key proteins such as NGF,TRKA,and GAP43 is involved in the nerve hyperplasia induced by mast cell degranulation.

The study explored the therapeutic effect and mechanism of the compound composed of tangerine peel,plum,agrimony and hawthorn on Eimeria tenella in chicks.In the experiment,144 chicks were divided into six groups:the blank group,the model group,the diclazuril group,and the high-dose,medium-dose and low-dose traditional Chinese medicine compound groups.The 14-day-old chickens were infected,the body weight and mortality were recorded,and the samples were dis-sected at 21 days of age.The anticoccidial effect of Chenpi compound at different doses was evalua-ted by calculating anticoccidial index,feed conversion rate and detecting serum cytokine levels and related mRNA expression levels.The results showed that the anticoccidial index of the middle dose group was 172.86,and the anticoccidial index of the low dose group was 158.98.In the middle and low dose groups,the levels of IL-1β,IL-6 and MDA in serum decreased significantly,while the levels of IL-4,IL10,T-AOC,SOD,CAT and GSH-Px increased significantly,and the mRNA ex-pression levels of related inflammation and antioxidant pathways changed.Further studies found that the medium-dose compound can regulate the ChTLR15/ChMyD88/ChNF-κB/ChNLRP3/Caspase-1 inflammatory signaling pathway,activate the Nrf2/Keap1/HO-1 antioxidant signaling pathway,and enhance the body's anti-inflammatory and antioxidant capacity;at the same time,it can increase the expression of intestinal tight junction ZO-1 and Occludin,repair the damaged in-testinal barrier,and play an anticoccidial role.The results showed that the middle dose of tangerine peel compound had a good effect in the treatment of coccidiosis,and its mechanism involved the regulation of anti-inflammatory and antioxidant pathways.

Objective To explore the molecular mechanisms by which cytochrome P4502C regulates ferroptosis in hu-man retinal microvascular endothelial cells(hRMECs)under high-glucose conditions.Methods Human retinal microvas-cular endothelial cells(hRMECs)were cultured in vitro and divided into six groups:the NG group(cells cultured in ser-um-free DMEM medium containing 5.5 mmol·L-1 glucose);the NG+fenofibrate group(10 μmol·L-1 fenofibrate added to the NG group);the NG+rifampicin group(10 μmol·L-1 rifampicin added to the NG group);the HG group(cells cul-tured in serum-free DMEM medium containing 25.5 mmol·L-1 glucose);the HG+fenofibrate group(10 μmol·L-1 feno-fibrate added to the HG group);and the HG+rifampicin group(10 μmol·L-1 rifampicin added to the HG group).After 48 hours of culture,subsequent experiments were conducted.Cell viability was assessed using the CCK-8 assay.The levels of malondialdehyde(MDA)and iron content in hRMECs were measured using commercial assay kits.The relative fluores-cence intensity of Liperfluo was quantified by flow cytometry.The mRNA expression levels of glutathione peroxidase 4(GPX4),cytochrome P4502C,and acyl-CoA synthetase long-chain family member 4(ACSL4)in hRMECs were analyzed using real-time quantitative PCR.The protein expression levels of GPX4,cytochrome P4502C,and ACSL4 in hRMECs were evaluated using Western blot analysis.Results Optimal concentration analysis showed that the 10 μmnol·L-1 fenofibrate group had the highest cell viability compared to the HG group(P＜0.05),while the 10 μmol·L-1 rifampicin group had the lowest cell viability(P＜0.05).Thus,10 μmol·L-1 was selected as the optimal concentration for both fenofibrate and rif-ampicin for subsequent experiments.Cell viability in the NG,NG+fenofibrate,NG+rifampicin,HG,HG+fenofibrate,and HG+rifampicin groups was(100.00±7.85)％,(102.15±9.07)％,(69.97±4.22)％,(61.74±6.13)％,(83.64±7.58)％,and(56.96±5.34)％,respectively.Compared with the NG group,the HG group had significantly lower cell via-bility(P＜0.05).Compared with the HG group,the HG+fenofibrate group had significantly higher cell viability(P＜0.05).Compared with the NG group,the HG group had higher levels of MDA,iron content,and Liperfluo relative fluores-cence intensity(all P＜0.05).Compared with the HG group,the HG+fenofibrate group had lower levels of MDA,iron content,and Liperfluo relative fluorescence intensity(all P＜0.05);the HG+rifampicin group had higher levels of these parameters(all P＜0.05).Compared with the NG group,the HG group had higher mRNA and protein expression levels of cytochrome P4502C and ACSL4,but lower levels of GPX4 mRNA and protein(all P＜0.05).Compared with the HG group,the HG+fenofibrate group had lower mRNA and protein expression levels of cytochrome P4502C and ACSL4,but higher lev-els of GPX4 mRNA and protein(all P＜0.05);the HG+rifampicin group had higher mRNA and protein expression levels of cytochrome P4502C and ACSL4,but lower levels of GPX4 mRNA and protein(all P＜0.05).Conclusion Cytochrome P4502C can promote ferroptosis of hRMECs under high-glucose conditions by regulating the GPX4/ACSL4 signaling axis;in-hibition of cytochrome P4502C expression can exert a protective effect on DR;modulation of the expression levels or bal-ance of the key factors GPX4 and ACSL4 in this pathway may hold promise as a potential strategy to slow the progression of DR.

Objective:To explore the distribution and clearance of 99mTc labeled diethylenetriamine pentaacetic acid(99mTc-DTPA)in different brain regions of adult rats after administration through brain extracellular space(ECS)pathway.Methods:After the injection of a volume of 2 μL and radioactive activity of about 3.7 MBq(100 μCi)of 99mTc-DTPA into the caudate nucleus and thalamus of SD rats through stereotactic positioning of rat brain,the single photon emission computed tomography/computed tomography(SPECT/CT)for small animals was used for imaging at different time points,and the dyna-mic distribution and clearance of the tracer in the whole body were observed continuously.The SD rats were injected with 99mTc-DTPA into thalamus and caudate nucleus respectively for biological distribution in vivo.They were put to death 4 h later.Their blood and urine were collected.The brain,cerebellum,heart,liver,spleen,lung,and kidney were taken and weighed by γ counter to measure its radioactivity.Results:SPECT/CT imaging results showed that after 99mTc-DTPA was administered through brain ECS,the radioactivity was concentrated in the brain,kidney and bladder.The tracer administered to the left caudate nucleus was preferentially drained to the right cerebellum,while the tracer administered to the right caudate nucleus was preferentially drained to the left cerebellum.There was a phenomenon of"con-tralateral cerebellar dominant drainage"in the caudate nucleus.The thalamic area preferentially drained to the ipsilateral cerebellum after administration.Four hours after administration via ECS,high radioac-tive uptake appeared in urine,cerebellum and brain,followed by blood and kidney.The radioactive up-take values of heart,liver,spleen and lung were low,which were mainly excreted through urinary sys-tem.Conclusion:Intracerebral ECS administration is a promising method of administration,but there are significant differences in distribution and clearance in different brain regions.This study further ex-pands the content and significance of"ECS regions",and also provides an important theoretical founda-tion for the treatment of encephalopathy and the research of new drugs through brain ECS in the future.

AIM:To establish a cell model of visceral hypersensitivity and nerve hyperplasia in irritable bowel syndrome(IBS)by conducting an in vitro co-culture of mouse P815 mast cells and N2a nerve cells and explore its possible mechanism.METHODS:Enzyme-linked immunosorbent assay(ELISA)with three replicates was used to confirm the C48/80-induced P815 degranulation.The length of neurites was observed under bright field microscopy to determine the number of differentiated neurons,thereby selecting the concentration of retinoic acid(RA)for stimulating the differentia-tion of N2a cells,with four replicates.A co-culture system of P815 and N2a cells was established using Transwell cham-bers with four replicates.The following groups were established:N2a cells cultured alone,N2a cells co-cultured with P815 cells,N2a cells co-cultured with P815 cells plus C48/80,and N2a cells plus RA group.After co-culturing,the num-ber of differentiated N2a cells was observed under bright field.The expression of nerve growth factor(NGF),tyrosine ki-nase receptor A(TRKA),growth-associated protein-43(GAP43),neuron-specific enolase(NSE),synapsin(SYN),and postsynaptic density protein-95(PSD-95)at both protein and gene levels in N2a cells was detected using Western blot and polymerase chain reaction(PCR),with four replicates.RESULTS:The best condition for N2a differentiation was stimulation with 10 μmol/L RA for 24 hours,whereas the best condition for degranulation was stimulation of P815 cells with 20 mg/L C48/80 for 24 hours.Compared with N2a cells cultured alone,the differentiation ratio of the N2a+P815+C48/80 and N2a+RA groups was significantly increased(P<0.01),and the protein and mRNA expressions of NGF,TRKA,GAP43,NSE,SYN,and PSD-95 were significantly increased(P<0.05).CONCLUSION:Our results revealed that mast cell degranulation enhances the level of nerve hyperplasia in enteric nerve cells and promotes changes in nerve structure and function.Synaptic remodeling regulated by abnormal expression of key proteins such as NGF,TRKA,and GAP43 is involved in the nerve hyperplasia induced by mast cell degranulation.