ObjectiveTo investigate the potential mechanism of Yiqi Yangyin Huoxue Formula （益气养阴活血方， YYHF） in the treatment of membranous nephropathy （MN） based on the NOX4-mediated stromal cell-derived factor-1α（SDF-1α）/CXC chemokine receptor 4 （CXCR4） signaling pathway. MethodsA total of 42 male SD rats were randomly divided into blank group （n=6） and modeling group （n=36）. The modeling rats were given a single injection of sheep antirat Fx1A （6 ml/kg） serum into the tail vein to establish MN model. Thirty-six rats with successful modeling were further divided into model group （normal saline， 10 ml/kg）， benazepril group （10 mg·kg^-1·d^-1）， the medium-dose YYHF group （11.0 g·kg^-1·d^-1） and high-dose YYHF group （22.0 g·kg^-1·d^-1）， with 9 rats in each group. All groups received daily intragastric administration for 6 consecutive weeks. During the experimental period， the general condition of rats in each group was observed. After gavage administration finished， a fully automated biochemical analyzer was used to measure the 24-hour urine total protein （24 h-UTP） and serum biochemical parameters， including albumin （ALB）， aspartate aminotransferase （AST）， blood urea nitrogen （BUN）， serum creatinine （SCr）， total cholesterol （TC）， and triglyceride （TG）. Hematoxylin-eosin （HE） staining and periodic acid-silver metheramine （PASM） staining were used to observe the histopathological changes of renal tissues in each group of rats. The levels of superoxide dismutase （SOD） and malondialdehyde （MDA） in renal tissues were detected by ELISA. The protein expression levels of NOX4， SDF-1α， CXCR4， Nrf2， nuclear factor kappa B subunit p65 （NF-κB p65） and podocyte slit diaphragm protein nephrin in rat renal tissues were detected by Western Blotting. The mRNA expressions of NOX4， SDF-1α， CXCR4， Nrf2， NF-κB p65 and Nephrin in renal tissues were detected by real-time fluorescence quantitative polymerase chain reaction （RT-PCR）. ResultsCompared to the blank group， rats in the model group exhibited typical features of MN， including lethargy， reduced activity， dull and coarse fur， marked edema， and massive proteinuria. Compared to the model group， these abnormal conditions were improved to varying degrees in all treatment groups. The high-dose YYHF group showed a level of improvement comparable to that of the benazepril group， with overall recovery superior to that of the medium-dose YYHF group. Compared to the blank group， the model group showed increased levels of 24 h-UTP， BUN， SCr， TC and TG， decreased level of ALB， increased levels of MDA and protein and mRNA expressions of NOX4， SDF-1α， CXCR4 and NF-κB p65 in renal tissue， along with decreased levels of SOD and protein and mRNA expressions of Nrf2， Nephrin （P＜0.01）， accompanied by severe renal pathological damage. Compared to the model group， the benazepril and the medium- and high-dose YYHF groups showed significant improvements in the above indicators （P＜0.01）， along with alleviation of renal pathological injury. The 24 h-UTP level in the high-dose YYHF group was lower than that in the medium-dose group （P＜0.01）， while no statistically significant difference was observed among the other treatment groups （P＞0.01）. ConclusionYYHF may alleviate oxidative stress and inflammatory responses by down-regulating the expression of NOX4 and inhibiting the SDF-1α/CXCR4-Nrf2 signaling pathway， thereby improving podocyte damage and protecting renal function.

Objective To explore the potential clinical value of the ratio of glycated albumin to albumin(GA/ALB)in the occurrence of heart failure(HF)among patients with coronary atherosclerotic heart disease(CHD).Methods A total of 337 CHD patients admitted to the Department of Cardiology in Shanxi Provincial People's Hospital from July 2023 to June 2024 were selected in this study.CHD patients were divided into HF group and non-HF group based on whether they progressed to HF.The clinical data and laboratory parame-ters of the two groups were compared.Restricted cubic spline curve was used to analyze the relationship be-tween GA/ALB levels and the risk of HF in CHD patients.Receiver operating characteristic curve was applied to evaluate the diagnostic efficacy of GA/ALB,GA,platelet to lymphocyte ratio(PLR),and monocyte to lym-phocyte ratio(MLR)in CHD patients with the occurrence of HF.Logistic regression was used to explore the relationship between serum GA/ALB levels and the risk of CHD patients occurrence of HF,and to analyze the degree of influence and stability of subgroup variables on results.Results There were statistically significant differences in GA/ALB,GA,PLR,MLR,and other indicators between the HF group and the non-HF group in CHD patients(P＜0.05).A non-linear relationship was observed between GA/ALB levels and the risk of HF in CHD patients.When the value of GA/ALB multiplied by 10 was less than 5.751,the risk of HF in CHD pa-tients increased with the increase of GA/ALB levels(P＜0.001).GA/ALB was an effective predictor for HF occurrence in CHD patients.Multivariable Logistic regression model showed that GA/ALB was an independ-ent risk factor for CHD patients with occurrence of HF.Subgroup analysis also confirmed the stability of GA/ALB in predicting the occurrence of HF in CHD patients.Conclusion GA/ALB is an independent risk factor for the occurrence of HF in CHD patients,and monitoring GA/ALB levels provides predictive value for the oc-currence of HF in these patients.

Hypoxia-related diseases,such as altitude sickness,acute lung injury,and heart failure,have emerged as significant public health challenges worldwide and are strongly associated with increased mortality risks.These conditions tend to cause functional impairment and structural alterations in multiple organs,primarily due to inadequate oxygen supply or the inability of tissues to utilize oxygen effectively.To elucidate the pathophysiological mechanisms underlying hypoxia-associated diseases and to explore potential therapeutic strategies,researchers have employed animal models to conduct extensive mechanistic studies and drug development trials.In this review,we provide a comprehensive overview of the methods used to construct animal models for various hypoxia-related diseases.We made a thorough evaluation of the strengths,weaknesses,limitations,and applicability of each approach.In particular,we focus on modeling strategies for key pathologies such as altitude sickness,acute lung injury,pulmonary hypertension,and heart failure,discussing their respective advantages and restrains.Furthermore,we examined the potential application of gene editing technologies in optimizing animal models,especially its role in enhancing the accuracy and reproducibility of the models.The integration of these advanced technologies holds great promise for developing animal models with improved specificity and simulation of key features of diseases,which will serve as a solid foundation for basic research,drug screening,and preclinical trials concerning hypoxia-related diseases.Herein,we present a systematic summary and future outlook of animal models for hypoxia-related diseases,ultimately providing theoretical support for developing clinical treatment strategies and improving patient outcomes.

Objective : To generate heterozygous TRAF2 knockout mice, the CRISPR/Cas9 technology was successfully employed. These mice were served as a valuable model to explore the pathological mechanisms underlying inflammatory and immune disorders mediated by abnormal TNF-α-TRAF2 signaling and to develop new therapeutic targets. Methods : A vector targeting the knockout of the TRAF2 gene was constructed. Lead RNA and Cas9 Mrna were introduced into the fertilized eggs of C57BL/6JGpt mice through microinjection to mediate the TRAF2 gene mutation in mice. The mouse tail protein was extracted and the genotype of the F0 generation was determined by PCR and Western blot. TRAF2+/- mice were successfully obtained. F0 generation mice were backcrossed with C57BL/6JGpt wild-type mice to obtain stable TRAF2+/- mice for propagation and subsequent experiments. The body weight of TRAF2+/- mice was detected; Western blot was used to detect the expression of TRAF2 in the spleen, liver and kidney tissues of TRAF2+/- mice. The development of spleen, liver and kidney tissues in TRAF2+/- mice was detected by HE staining. Results : PCR identification using specific primers demonstrated that TRAF2+/- mice exhibited a target band at 679 bp. Western blot analysis results indicated that, compared with the WT group, the expression of TRAF2 in the tail protein of TRAF2+/- mice was significantly reduced(P+/- mice had a lower body weight compared to their littermate WT mice(P+/- mice was decreased(P+/- mice and WT mice. Conclusion The successful construction of TRAF2+/- mice has provided an important animal model for exploring the role of TRAF2 in developmental regulation, revealing the mechanism of inflammatory immune diseases mediated by abnormal TNF-α-TRAF2 signaling, and screening related drug targets.

Objective:To investigate the role of rheum tanguticum polysaccharide(RTP)in treating ulcerative colitis(UC)by improving intestinal flora.Methods:UC model was established,rats were divided into normal control group,model group and RTP treatment group.The integrity of colonic mucosa,submucosa and muscularis were observed by HE staining.Expressions of TNF-α,IL-6,IL-8 and IL-1β in serum were assessed by ELISA.16S rRNA sequencing was performed to analyze the composition and differences of intestinal flora in rats.Results:UC rats had symptoms such as loose stools,blood in the stool,anal filth,chills and mental malaise.RTP treatment could significantly improve the symptoms and reduce DAI scores(P<0.05).HE staining showed that the colonic struc-ture of UC rats was necrotic and inflammatory cells infiltrated submucosa.RTP treatment could significantly promote the repair and regeneration of mucosal epithelium.RTP could significantly inhibit inflammatory response and downregulate the expression of inflam-matory factors in UC rats(P<0.05).Composition and abundance of intestinal flora in UC rats were significantly different from that of normal control group.Staphylococcus was significantly increased,while Collinsella,Allobaculum,Chlamydia and Holdemanella were significantly decreased.RTP treatment could improve intestinal flora,reduce the abundance of Staphylococcus,and increase the abundance of Collinsella,Allobaculum,Chlamydia and Holdemanella.Conclusion:RTP can be used to treat UC in rats by inhibiting inflammation and improving intestinal flora.

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 explore the effect of scalp electroacupuncture (EA) on central pain sensitization in rats with knee osteoarthritis (KOA).Methods:Thirty-two 8-week-old female Sprague-Dawley rats were randomly divided into a blank control group, a model group, an electroacupuncture (EA) group and a sham EA group, each of 8. All of the rats except those in the control group had KOA induced through intra-articular monosodium iodoacetate injections in the right knee. Two weeks later the EA group rats began receiving daily head EA sessions 6 days/week for 2 weeks. The sham EA group received identical but non-therapeutic stimulation. The blank control and model groups received no EA intervention. Before the modelling and 3, 7, 14, 21 and 28 days later, all of the rats completed bipedal balance pain tests and mechanical allodynia evaluations. After the testing on day 28, all of the rats were euthanized for molecular analyses. Western blotting and immunohistochemistry were performed to examine protein expression of brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and cannabinoid receptor type 1 (CB1R) in both the periaqueductal gray (PAG) matter and spinal dorsal horns (SDHs). Serum levels of substance P (SP) and 5-hydroxytryptamine (5-HT) were also quantified using enzyme-linked immunosorbent assays.Results:Three days after successful modeling, the average weight-bearing capacity of the right hind limb in the model, sham EA and EA groups was significantly lower than that of the blank controls. It reached its lowest level on the 14th day after modeling. Concurrently, the pain responses in those three groups were significantly higher than among the controls, also peaking on the 14th day after modeling. After two weeks of electroacupuncture, the electroacupuncture group showed significant improvement in both right hind limb weight-bearing capacity and pain response compared to the model group. Meanwhile, the levels of BDNF and TrkB protein in the periaqueductal gray matter were significantly higher in the model group than among the blank controls, while the electroacupuncture group exhibited significantly reduced expression of BDNF and TrkB proteins compared to the model group, along with significantly increased CB1R protein expression. The model group showed significantly elevated expression of both BDNF and TrkB proteins in the spinal dorsal horn compared to the blank control group, while there were significant differences between the EA and model groups in the expression of BDNF, TrkB and CB1R proteins. Immunohistochemical analysis on day 28 revealed that the EA group had significantly fewer BDNF- and TrkB-positive cells in the PAG compared to the model group, with significantly more CB1R-positive cells. In the SDH, the model group exhibited significantly increased numbers of BDNF- and TrkB-positive cells compared to the blank control group, whereas significant differences were found between the EA and blank control groups in the numbers of BDNF-, TrkB- and CB1R-positive cells. Serum analysis on day 28 demonstrated that substance P and 5-hydroxytryptamine levels in the model, sham EA and EA groups were significantly higher than in the blank control group, on average. However, no significant differences were observed in serum SP and 5-HT levels between the EA and model groups.Conclusions:Scalp EA significantly alleviates central pain sensitization in KOA, at least in rats, potentially by suppressing BDNF and TrkB expression while upregulating CB1R expression in the PAG matter and the SDH.

Objective To explore the therapeutic effect and mechanism of electroacupuncture on depressive model mice based on the phosphatase and tensin homolog-induced kinase 1(PINK1)/Parkin pathway.Methods Specific pathogen-free grade male C57BL/6J mice were used.For experiment 1,60 mice were randomly divided into blank,model,sham electroacupuncture,and electroacupuncture groups using the random number table method,with 15 rats per group.For experiment 2,30 mice were randomly divided into normal,cyclosporine A(CsA),and electroacupuncture+CsA groups using the same method,with 10 rats per group.The chronic restraint stress(CRS)was used to establish a depression model.After successful modeling,CRS was continued to maintain model stability.After modeling,1 h before daily CRS stimulation,the electroacupuncture and electroacupuncture+CsA groups received electroacupuncture interventions at the"Baihui"(GV20)and"Zusanli"(ST36)acupoints,using continuous wave stimulation at a frequency of 2 Hz and an intensity of 1 mA for 20 min,once daily for 7 consecutive days.Mice in the sham electroacupuncture group received superficial needling at non-meridian,non-acupoint locations under the axilla 1 h before CRS,with the electroacupuncture device connected but not powered on once a day for 7 consecutive days.Mice in the CsA and electroacupuncture+CsA groups received an intraperitoneal injection of CsA solution(0.2 mg/g)30 min before electroacupuncture intervention,once daily for 7 consecutive days.In experiment 1,depressive-like behavior was assessed using the open field,tail suspension,and sucrose preference tests.The spontaneous excitatory postsynaptic currents(sEPSC)and spontaneous inhibitory postsynaptic currents(sIPSC)parameters of hippocampal neurons were evaluated using brain slice patch clamp techniques.Western blotting was conducted to measure the expression levels of mitochondrial autophagy-related proteins PINK 1,phosphorylated PINK1(p-PINK1),Parkin,microtubule-associated protein 1A/1B-light chain 3-Ⅱ/Ⅰ(LC3-Ⅱ/Ⅰ),ubiquitin-binding protein(p62),and mitochondrial markers,including translocase of outer mitochondrial membrane 20(TOMM20),heat shock protein 60(HSP 60),and cytochrome c oxidase Ⅳ(COX Ⅳ).Immunofluorescence was used to detect PINK1 protein expression in the CA3 region of the hippocampus.Transmission electron microscopy was used to examine the ultrastructure of mitochondria in hippocampal neurons.On the basis of experiment 1,experiment 2 evaluated depressive-like behavior in mice using sucrose preference,open field,and tail suspension tests;Western blotting was used to detect the expressions of PINK1,p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,p62,TOMM20,HSP 60,and COX Ⅳ proteins of hippocampus in mice.The mitochondrial ultrastructure was observed in hippocampal neurons using transmission electron microscopy.Results In experiment 1,compared with the blank group,the model and sham electroacupuncture groups exhibited a decrease in sucrose consumption rate,a decrease in the time spent in the center area,a reduced proportion of distance moved in the center area,and an increase in immobility time of tail suspension(P＜0.05).The sEPSC and sIPSC in hippocampal neurons decreased in both amplitude and frequency(P＜0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COXⅣ expression levels in the hippocampus were reduced,whereas the p62 expression level was increased(P＜0.05).The average fluorescence intensity of PINK1 in the CA3 region of the hippocampus decreased(P＜0.05).The number of healthy mitochondria in hippocampal neurons was reduced,with numerous damaged mitochondrial structures observed.Compared to the model and sham electroacupuncture groups,the electroacupuncture group showed an increased in the time spent in the center area,a higher proportion of distance moved in the center area,and an elevated sucrose consumption rate.In contrast,the immobility time in the tail suspension test decreased(P＜0.05),whereas the amplitude and frequency of sEPSC and sIPSC in hippocampal neurons increased(P＜0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COXⅣ expression levels in the hippocampus increased,whereas the p62 expression level decreased(P＜0.05).The average fluorescence intensity of PINK1 in the CA3 region of the hippocampus increased(P＜0.05).Additionally,mitochondrial damage in hippocampal neurons was alleviated,and a notable presence of autophagosomes mitophagy lysosomes was observed.In experiment 2,compared with the normal group,the mice in the CsA and electroacupuncture+CsA groups showed a decrease in the time spent in the center area and the proportion of distance moved in the center area,a decrease in sucrose consumption rate,and an increase in the immobility time in the tail suspension test(P＜0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COX Ⅳ expression levels in the hippocampus decreased,whereas p62 expression increased(P＜0.05).Many damaged mitochondria were observed in hippocampal neurons.Conclusion Electroacupuncture may exert its antidepressant effects by promoting PINK1/Parkin pathway-mediated mitophagy to eliminate damaged mitochondria,thereby restoring the function of hippocampal neurons in depressive model mice.

In order to investigate the effect of SIRT2 on tight junctions and endoplasmic reticulum stress in liver tissues of cold-treated mice,10 each of 5-week-old male C57BL/6 mice and SIRT2 knockout mice were selected and randomly divided into the wild-type room-temperature control group(WT Control),the wild-type cold-treated group(WT Cold),the SIRT2 knockout+room-temperature control group(KO Control)and SIRT2 knockout+cold treatment group(KO Cold).Mice in the room-temperature control group were kept at a temperature of(24+2)℃,and the cold-treatment group was placed in a(4+2)℃ artificial climate chamber for 3 h of random stimu-lation per day for 3 weeks.H&E staining,Masson staining,and transmission electron microscopy were employed to examine the microscopic and ultrastructural changes in mouse liver;AST and ALT concentrations in mouse serum were detected by biochemical analyzers;Western blot analysis was used to detect the expressions of tight junction-related proteins(Claudin1,Occludin),endo-plasmic reticulum stress-related proteins(GRP78,CHOP,XBP1,p-eIF2α,eIF2α),and pro-inflam-matory cytokines(TNF-α,IL-1β,IL-6).The results showed that compared with WT Control,the liver lobular structure of WT Cold and KO Control mice was unclear,hepatic cord arrangement was disordered,cytoplasm was loose,white vacuoles appeared,a small amount of collagen deposi-tion and fibroplasia,mitochondria were slightly swollen in hepatocytes,and endoplasmic reticulum was unevenly distributed,while the serum concentrations of AST and ALT were increased(P＜0.050,P＜0.010),and the liver tissues showed decreased protein expression of Occludin and Clau-din1(P＜0.050,P＜0.010,P＜0.001),and increased protein expression of GRP78,CHOP,XBP1,p-eIF2α/eIF2α,TNF-α,IL-6,and IL-1β(P＜0.050,P＜0.010,P＜0.001);compared with the KO Control,KO Cold mice showed a large number of white vacuoles,a small number of balloon-like lesions,inflammatory cell infiltration,obvious collagen deposition and fibroplasia,mitochondrial swelling in hepatocytes,mitochondrial ridge reduction,endoplasmic reticulum thickening,and ser-um AST and ALT concentrations increased(P＜0.010),and in liver tissue,the protein expression of Occludin and Claudinl decreased(P＜0.010),while the protein expression of GRP78,CHOP,XBP1,p-eIF2α/eIF2α,TNF-α,IL-6 and IL-1β increased(P＜0.050,P＜0.010,P＜0.001).The re-sults showed that SIRT2 knockdown could aggravate the liver tissue tight junction damage caused by cold treatment,induce endoplasmic reticulum stress,and further promotes the inflammatory re-sponse.

Enamel demineralization, the formation of white spot lesions, is a common issue in clinical orthodontic treatment. The appearance of white spot lesions not only affects the texture and health of dental hard tissues but also impacts the health and aesthetics of teeth after orthodontic treatment. The prevention, diagnosis, and treatment of white spot lesions that occur throughout the orthodontic treatment process involve multiple dental specialties. This expert consensus will focus on providing guiding opinions on the management and prevention of white spot lesions during orthodontic treatment, advocating for proactive prevention, early detection, timely treatment, scientific follow-up, and multidisciplinary management of white spot lesions throughout the orthodontic process, thereby maintaining the dental health of patients during orthodontic treatment.
Humans ; Consensus ; Dental Caries/etiology* ; Dental Enamel/pathology* ; Tooth Demineralization/etiology* ; Tooth Remineralization