ObjectiveTo explore the mechanism of ventricular remodeling mediated by the p38 mitogen-activated protein kinase （MAPK）/nuclear factor kappa B （NF-κB） signaling pathway in the rat model of chronic heart failure （CHF） with heart-blood stasis syndrome， as well as the intervention effect of Danhong injection. MethodsIn vivo experiment： SPF-grade male SD rats were assigned via the random number table method into 4 groups： Sham operation， model， captopril （8.8 mg·kg^-1）， and Danhong injection （6.0 mL·kg^-1）. The model of CHF with heart-blood stasis syndrome was established by abdominal aortic constriction， and the sham operation group only underwent laparotomy without constriction. All the groups were treated continuously for 15 days. The tongue color of rats was observed. Echocardiography， hemorheology， heart mass index （HMI）， and left ventricular mass index （LVMI） were measured. Hematoxylin-eosin （HE） staining and Masson staining were performed to observe the pathological and fibrotic changes of the myocardial tissue. Enzyme-linked immunosorbent assay （ELISA） was employed to quantify the levels of N-terminal pro-B-type natriuretic peptide （NT-proBNP）， interleukin-6 （IL-6）， angiotensin Ⅱ （AngⅡ）， tumor necrosis factor-α （TNF-α）， and Creactive protein （CRP） in the serum， as well as the levels of matrix metalloproteinase-2 （MMP-2） and matrix metalloproteinase-9 （MMP-9） in the myocardial tissue. Western blot was used to quantify the protein levels of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 in the myocardial tissue. In vitro experiment： H9C2 cardiomyocytes were treated with 1×10^-6 mol·L^-1 AngⅡ to establish a model of myocardial hypertrophy. H9C2 cardiomyocytes were allocated into normal， model， inhibitor + Danhong injection， Danhong injection （20 mL·L^-1）， and inhibitor （SB203580， 5 μmol·L^-1） groups. CCK-8 assay was used to detect the viability of H9C2 cardiomyocytes. Rhodamine-labeled phalloidin staining was used to reveal the area of cardiomyocytes. Real-time PCR was performed to determine the mRNA levels of atrial natriuretic peptide （ANP） and brain natriuretic peptide （BNP）. Western blot was used to assess the protein levels of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65. ResultsIn vivo experiment： Compared with the sham operation group， the model group showed purplish-dark tongue with decreased R， G， B values of the tongue surface （P<0.01）， increased whole blood viscosity （at low， medium， and high shear rates） （P<0.01）， decreased left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） （P<0.01）， increased left ventricular end-diastolic diameter （LVIDd）， left ventricular end-systolic diameter （LVIDs）， and left ventricular posterior wall thickness at end-diastole （LVPWd） （P<0.01）， raised LVMI and HMI （P<0.01）， and elevated levels of NT-proBNP， TNF-α， IL-6， and CRP in the serum and MMP-2 and MMP-9 in the myocardial tissue （P<0.01）. The HE and Masson staining of the myocardial tissue showed compensatory myocardial hypertrophy， fibrosis， and massive inflammatory cell infiltration in the model group. Additionally， the model group presented up-regulated protein levels of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 in the myocardial tissue （P<0.01）. Compared with the model group， each administration group showed increased R， G， B values of the tongue surface （P<0.05， P<0.01）， decreased whole blood viscosity （at low， medium， and high shear rates） （P<0.05， P<0.01）， increased LVEF and LVFS （P<0.01）， decreased LVIDd， LVIDs， and LVPWd （P<0.05， P<0.01）， declined LVMI and HMI （P<0.05， P<0.01）， and lowered levels of NT-proBNP， TNF-α， IL-6， and CRP in the serum and MMP-2 and MMP-9 in the myocardial tissue （P<0.01）. HE and Masson staining showed alleviated compensatory myocardial hypertrophy， reduced fibrosis， and decreased expression of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 in the myocardial tissue （P<0.01）. In vitro experiment： When the concentration of Danhong injection reached 20 mL·L^-1， the survival rate of H9C2 cardiomyocytes was the highest （P<0.01）. Compared with the normal group， the model group showed up-regulated mRNA levels of ANP and BNP （P<0.01）， increased relative cell surface area （P<0.01）， and raised protein levels of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 （P<0.01）. Compared with the model group， each administration group showed down-regulated mRNA levels of ANP and BNP （P<0.01）， reduced relative cell surface area （P<0.05， P<0.01）， and down-regulated protein levels of p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 （P<0.05， P<0.01）. ConclusionDanhong injection can regulate ventricular remodeling through the p38 MAPK/NF-κB pathway， thereby exerting a protective effect on the rat model of CHF with heart-blood stasis syndrome.

Objective:To compare the mortality of patients with septic shock in the intensive care unit of the emergency department of Beijing Chaoyang Hospital in 2012 and 2022, and analyze the risk factors affecting the prognosis of patients in each year.Methods:According to the diagnostic criteria of septic shock, 82 patients with septic shock admitted to the ICU of the emergency department of Beijing Chaoyang Hospital in 2012 and 52 patients with septic shock admitted to the ICU in 2022 were included. The clinical data of patients in each year and the related indicators that may affect the prognosis were compared. The risk factors of death in patients with septic shock in each year were screened by multivariate logistic regression analysis, and the predictive value of risk factors on death was evaluated by receiver operating characteristic (ROC) curve.Results:In 2012, 30 patients with septic shock died and 52 survived. The fatality rate was 36.59%. In 2022, 16 patients with septic shock died and 36 survived, with a fatality rate of 30.77%. There was significant difference in mortality between 2012 and 2022 (χ 2=6.805, P=0.009). In 2012 and 2022, the mortality of septic shock patients with different gender, age and Sequential Organ failure assessment (SOFA) score had statistical significance ( P<0.05). Multivariate logistic regression analysis showed that gender ( OR=1.554, P=0.037), lactic acid ( OR=1.062, P=0.035) and SOFA score ( OR=1.199, P=0.028) were the risk factors affecting the prognosis of patients in 2012, gender ( OR=1.234, P=0.028), total cholesterol ( OR=1.358, P=0.028) and SOFA score ( OR=1.388, P=0.034) were the risk factors affecting the prognosis of patients in 2022. ROC curve analysis results showed that SOFA score had higher sensitivity and specificity in predicting death of septic shock patients in 2012 (all P<0.05), and lactic acid, total cholesterol and SOFA score had higher sensitivity and specificity in predicting death of septic shock patients in 2022 (all P<0.05). Conclusions:The case fatality rate of septic shock patients in 2022 is lower than that in 2012, the morbidity and mortality rate of male patients are still higher than that of female patients, and the case fatality rate of patients increases with age. SOFA score was an independent risk factor for the prognosis of septic shock patients in 2012 and 2022.

Objective:To compare the differential regulatory effects of corticotropin releasing hormone(CRH)neu-rons in periaqueductal gray(PAG)and medial preoptic area(MPA)downstream of paraventricular nucleus(PVN)in mediating stress-related abnormal behaviors.Methods:The anterograde labeled virus AAV2/9-hSyn-DIO-hChR2-EYFP and AAV2/9-mCrh-SV40 NLS-Cre was injected into the PVN brain region of mouse,and the projection distribution of PVN axons in the PAG and MPA brain regions was observed after statistical analysis of its projection distribution through Image J.The optogenetic inhibitory virus mixture of AAV2/9-DIO-stGtACR2-EGFP and AAV2/9-mCrh-SV40 NLS-Cre targeting CRH neurons was injected into the PAG and MPA brain regions receiving the corresponding PVN projection,respectively,and the CRH neurons in the PAG and MPA brain regions were inhibited by optogenetic blue light 460 nm to observe the anxiety-related behaviors of mice under acute restraint stress.Results:Densely distributed CRHergic PVN axon terminals were observed in PAG and MPA brain regions.Optogenetic inhibition of CRH neurons in the PAG region of acute stress mice showed no significant change in social preference behavior.The eating latency decreased in the novelty-suppressed feeding test,and the escape latency increased under visual fear stimulation.Optogenetic inhibi-tion of CRH neurons in the MPA brain region showed no significant change in social preference,significantly decreased eating latency in the novelty-suppressed feeding test,and no significant change in escape latency under visual fear stim-ulation.Conclusion:CRH neurons in PAG and MPA brain regions downstream of PVN have differential regulation in a-cute stress-related anxiety behavior,but no difference in social behavior regulation,which provides theoretical support and basis for in-depth exploration of stress-related brain regions and cellular mechanisms.

Objective To investigate the effects of inclined axial compressive force and flexion moment on the anterior and posterior shear stiffness of the lumbosacral segment.Methods Six fresh-frozen human cadaveric L5-S1 segments were tested under intact and two progressively impaired structural conditions:intact,a 4-mm bilateral facet joint gap,and anterior discectomy with nucleus pulposus removal plus circumferential release of the inner annular fibers(disc injury).A 300 N axial compressive force was applied either vertically downward or with a 10° or 20° anterior inclination through the disc's shear center.Anterior(0 N to 250 N)and posterior(-50 N to 0 N)shear tests were conducted using a material testing machine.These tests were repeated under a 5 N-m flexion moment.The relative motion between L5 and Si was measured using a three-dimensional motion capture system.Results In the intact state,the inclination of the axial compressive force did not significantly alter anterior or posterior shear stiffness.However,the application of a flexion moment increased anterior shear stiffness by 49.3％.Progressive structural damage resulted in incremental increases in anteroposterior shear translation and corresponding reductions in stiffness.Notably,under combined loading with axial compression and flexion moment,anterior stiffness decreased from 939 N/mm(intact)to 224 N/mm(disc injury),while posterior stiffness decreased from 572 N/mm to 217 N/mm.Within the low-load range,no significant differences in shear stiffness were observed across any structural conditions,regardless of axial force inclination or combined with a flexion moment.Conclusions This study supports the clinical view that retro-inclination of the pelvis serves as a compensatory mechanism to enhance segmental shear stability.However,this compensatory capacity gradually diminishes and ultimately fails as spinal degeneration progresses.

Objective:To compare the differential regulatory effects of corticotropin releasing hormone(CRH)neu-rons in periaqueductal gray(PAG)and medial preoptic area(MPA)downstream of paraventricular nucleus(PVN)in mediating stress-related abnormal behaviors.Methods:The anterograde labeled virus AAV2/9-hSyn-DIO-hChR2-EYFP and AAV2/9-mCrh-SV40 NLS-Cre was injected into the PVN brain region of mouse,and the projection distribution of PVN axons in the PAG and MPA brain regions was observed after statistical analysis of its projection distribution through Image J.The optogenetic inhibitory virus mixture of AAV2/9-DIO-stGtACR2-EGFP and AAV2/9-mCrh-SV40 NLS-Cre targeting CRH neurons was injected into the PAG and MPA brain regions receiving the corresponding PVN projection,respectively,and the CRH neurons in the PAG and MPA brain regions were inhibited by optogenetic blue light 460 nm to observe the anxiety-related behaviors of mice under acute restraint stress.Results:Densely distributed CRHergic PVN axon terminals were observed in PAG and MPA brain regions.Optogenetic inhibition of CRH neurons in the PAG region of acute stress mice showed no significant change in social preference behavior.The eating latency decreased in the novelty-suppressed feeding test,and the escape latency increased under visual fear stimulation.Optogenetic inhibi-tion of CRH neurons in the MPA brain region showed no significant change in social preference,significantly decreased eating latency in the novelty-suppressed feeding test,and no significant change in escape latency under visual fear stim-ulation.Conclusion:CRH neurons in PAG and MPA brain regions downstream of PVN have differential regulation in a-cute stress-related anxiety behavior,but no difference in social behavior regulation,which provides theoretical support and basis for in-depth exploration of stress-related brain regions and cellular mechanisms.

Objective To investigate the effects of inclined axial compressive force and flexion moment on the anterior and posterior shear stiffness of the lumbosacral segment.Methods Six fresh-frozen human cadaveric L5-S1 segments were tested under intact and two progressively impaired structural conditions:intact,a 4-mm bilateral facet joint gap,and anterior discectomy with nucleus pulposus removal plus circumferential release of the inner annular fibers(disc injury).A 300 N axial compressive force was applied either vertically downward or with a 10° or 20° anterior inclination through the disc's shear center.Anterior(0 N to 250 N)and posterior(-50 N to 0 N)shear tests were conducted using a material testing machine.These tests were repeated under a 5 N-m flexion moment.The relative motion between L5 and Si was measured using a three-dimensional motion capture system.Results In the intact state,the inclination of the axial compressive force did not significantly alter anterior or posterior shear stiffness.However,the application of a flexion moment increased anterior shear stiffness by 49.3％.Progressive structural damage resulted in incremental increases in anteroposterior shear translation and corresponding reductions in stiffness.Notably,under combined loading with axial compression and flexion moment,anterior stiffness decreased from 939 N/mm(intact)to 224 N/mm(disc injury),while posterior stiffness decreased from 572 N/mm to 217 N/mm.Within the low-load range,no significant differences in shear stiffness were observed across any structural conditions,regardless of axial force inclination or combined with a flexion moment.Conclusions This study supports the clinical view that retro-inclination of the pelvis serves as a compensatory mechanism to enhance segmental shear stability.However,this compensatory capacity gradually diminishes and ultimately fails as spinal degeneration progresses.

Objective:To compare the mortality of patients with septic shock in the intensive care unit of the emergency department of Beijing Chaoyang Hospital in 2012 and 2022, and analyze the risk factors affecting the prognosis of patients in each year.Methods:According to the diagnostic criteria of septic shock, 82 patients with septic shock admitted to the ICU of the emergency department of Beijing Chaoyang Hospital in 2012 and 52 patients with septic shock admitted to the ICU in 2022 were included. The clinical data of patients in each year and the related indicators that may affect the prognosis were compared. The risk factors of death in patients with septic shock in each year were screened by multivariate logistic regression analysis, and the predictive value of risk factors on death was evaluated by receiver operating characteristic (ROC) curve.Results:In 2012, 30 patients with septic shock died and 52 survived. The fatality rate was 36.59%. In 2022, 16 patients with septic shock died and 36 survived, with a fatality rate of 30.77%. There was significant difference in mortality between 2012 and 2022 (χ 2=6.805, P=0.009). In 2012 and 2022, the mortality of septic shock patients with different gender, age and Sequential Organ failure assessment (SOFA) score had statistical significance ( P<0.05). Multivariate logistic regression analysis showed that gender ( OR=1.554, P=0.037), lactic acid ( OR=1.062, P=0.035) and SOFA score ( OR=1.199, P=0.028) were the risk factors affecting the prognosis of patients in 2012, gender ( OR=1.234, P=0.028), total cholesterol ( OR=1.358, P=0.028) and SOFA score ( OR=1.388, P=0.034) were the risk factors affecting the prognosis of patients in 2022. ROC curve analysis results showed that SOFA score had higher sensitivity and specificity in predicting death of septic shock patients in 2012 (all P<0.05), and lactic acid, total cholesterol and SOFA score had higher sensitivity and specificity in predicting death of septic shock patients in 2022 (all P<0.05). Conclusions:The case fatality rate of septic shock patients in 2022 is lower than that in 2012, the morbidity and mortality rate of male patients are still higher than that of female patients, and the case fatality rate of patients increases with age. SOFA score was an independent risk factor for the prognosis of septic shock patients in 2012 and 2022.

Objective: To investigate the mechanism of Yishen Tongluo Formula in ameliorating renal pyroptosis in diabetic nephropathy mice by regulating the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. Methods: Sixty C57BL/6 male mice were randomly divided into control (10 mice) and intervention groups (50 mice) using random number table method. The diabetes nephropathy model was established by intraperitoneally injecting streptozotocin(50 mg/kg). After modeling, the intervention group was further divided into model, semaglutide (40 μg/kg), and high-, medium-, and low-dose Yishen Tongluo Formula groups (15.6, 7.8, and 3.9 g/kg, respectively) using random number table method. The high-, medium-, and low-dose Yishen Tongluo Formula groups were administered corresponding doses of medication by gavage, the semaglutide group received a subcutaneous injection of semaglutide injection, and the control group and model groups were administered distilled water by gavage for 12 consecutive weeks. Random blood glucose levels of mice in each group were monitored, and the 24-h urinary protein content was measured using biochemical method every 4 weeks; after treatment, the serum creatinine and urea nitrogen levels were measured using biochemical method. The weight of the kidneys was measured, and the renal index was calculated. Hematoxylin and eosin, periodic acid-Schiff, periodic Schiff-methenamine, and Masson staining were used to observe the pathological changes in renal tissue. An enzyme-linked immunosorbent assay was used to detect urinary β2-microglobulin (β2-MG), neutrophil gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1) levels. Western blotting and real-time fluorescence PCR were used to detect the relative protein and mRNA expression levels of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3), Caspase-1, gasdermin D (GSDMD), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in renal tissue. Immunohistochemistry was used to detect the proportion of protein staining area of the TLR4, MyD88, and NF-κB in renal tissue. Results: Compared with the control group, the random blood glucose, 24-h urinary protein, serum creatinine, urea nitrogen, and renal index of the model group increased, and the urine β2-MG, NGAL, and KIM-1 levels increased. The relative protein and mRNA expression levels of NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 in renal tissue increased, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas increased (P<0.05). Pathological changes such as glomerular hypertrophy were observed in the renal tissue of the model group. Compared with the model group, the Yishen Tongluo Formula high-dose group showed a decrease in random blood glucose after 12 weeks of treatment (P<0.05). The Yishen Tongluo Formula high- and medium-dose groups showed a decrease in 24-h urinary protein, creatinine, urea nitrogen, and renal index, as well as decreased β2-MG, NGAL, and KIM-1 levels. NLRP3, Caspase-1, GSDMD, IL-1 β, and IL-18 relative protein and mRNA expression levels were also reduced, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas was reduced (P<0.05). Pathological damage to renal tissue was ameliorated. Conclusion Yishen Tongluo Formula may exert protective renal effects by inhibiting renal pyroptosis and alleviating tubular interstitial injury in diabetic nephropathy mice by regulating the TLR4/MyD88/NF-κB signaling pathway.

ObjectiveTo investigate the effect and mechanism of Yishen Tongluo prescription in protecting mice from oxidative stress injury in diabetic kidney disease （DKD） via the nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）/NAD（P）H quinone oxidoreductase 1 （NQO1） signaling pathway. MethodsSpecific pathogen-free （SPF） male C57BL/6 mice were assigned into a control group （n=10） and a modeling group （n=50）. The DKD model was established by intraperitoneal injection of streptozotocin. The mice in the modeling group were randomized into a model group， a semaglutide （40 μg·kg^-1） group， and high-， medium-， and low-dose （18.2， 9.1， 4.55 g·kg^-1， respectively） Yishen Tongluo prescription groups， with 10 mice in each group. The treatment lasted for 12 weeks. Blood glucose and 24-h urine protein levels were measured， and the kidney index （KI） was calculated. Serum levels of creatinine （SCr）， blood urea nitrogen （BUN）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST） were assessed. The pathological changes in the renal tissue were evaluated by hematoxylin-eosin， periodic acid-Schiff， periodic acid-silver methenamine， and Masson’s trichrome staining. Enzyme-linked immunosorbent assay kits were used to measure the levels of β2-microglobulin （β2-MG）， neutrophil gelatinase-associated lipocalin （NGAL）， kidney injury molecule-1 （KIM-1）， liver fatty acid-binding protein （L-FABP）， nitric oxide synthase （NOS）， glutathione （GSH）， total antioxidant capacity （T-AOC）， and 8-hydroxy-2'-deoxyguanosine （8-OHdG）. Immunohistochemical staining was performed to examine the expression of Kelch-like ECH-associated protein 1 （Keap1） and malondialdehyde （MDA）. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR） and Western blot were employed to determine the mRNA and protein levels， respectively， of factors in the Nrf2/HO-1/NQO1 signaling pathway. ResultsCompared with the control group， the DKD model group showed rises in blood glucose， 24-h urine protein， KI， SCr， BUN， and ALT levels， along with glomerular hypertrophy， renal tubular dilation， thickened basement membrane， mesangial expansion， and collagen deposition. Additionally， the model group showed elevated levels of β2-MG， NGAL， KIM-1， L-FABP， NOS， and 8-OHdG， lowered levels of GSH and T-AOC， up-regulated expression of MDA and Keap1， and down-regulated expression of Nrf2， HO-1， NQO1， and glutamate-cysteine ligase catalytic subunit （GCLC） （P<0.05）. Compared with the model group， the semaglutide group and the medium- and high-dose Yishen Tongluo prescription groups showed reductions in blood glucose， 24-h urine protein， KI， SCr， BUN， and ALT levels， along with alleviated pathological injuries in the renal tissue. In addition， the three groups showed lowered levels of β2-MG， NGAL， KIM-1， L-FABP， NOS， and 8-OHdG， elevated levels of GSH and T-AOC， down-regulated expression of MDA and Keap1， and up-regulated expression of Nrf2， HO-1， NQO1， and GCLC （P<0.05）. ConclusionYishen Tongluo prescription exerts renoprotective effects in the mouse model of DKD by modulating the Nrf2/HO-1/NQO1 signaling pathway， mitigating oxidative stress， and reducing renal tubular injuries.

Chronic pain, frequently comorbid with neuropsychiatric disorders, significantly impairs patients' quality of life and functional capacity. Accumulating evidence implicates the chemokine CCL2 and its receptor CCR2 as key players in chronic pain pathogenesis. This review examines the regulatory mechanisms of the CCL2/CCR2 axis in chronic pain processing at three hierarchical levels: (1) Peripheral Sensitization: CCL2/CCR2 modulates TRPV1, Nav1.8, and HCN2 channels to increase neuronal excitability and CGRP signaling and calcium-dependent exocytosis in peripheral nociceptors to transmit pain. (2) Spinal Cord Central Sensitization: CCL2/CCR2 contributes to NMDAR-dependent plasticity, glial activation, GABAergic disinhibition, and opioid receptor desensitization. (3) Supraspinal Central Networks: CCL2/CCR2 signaling axis mediates the comorbidity mechanisms of pain with anxiety and cognitive impairment within brain regions, including the ACC, CeA, NAc, and hippocampus, and it also increases pain sensitization through the descending facilitation system. Current CCL2/CCR2-targeted therapeutic strategies and their development status are discussed, highlighting novel avenues for chronic pain management.
Humans ; Chronic Pain/physiopathology* ; Animals ; Neuroglia/metabolism* ; Chemokine CCL2/metabolism* ; Receptors, CCR2/metabolism*