OBJECTIVE: To investigate the anti-inflammatory effect of rutaecarpine (RUT) on monosodium urate crystal (MSU)-induced murine peritonitis in mice and further explored the underlying mechanism of RUT in lipopolysaccharide (LPS)/MSU-induced gout model in vitro. METHODS: In MSU-induced mice, 36 male C57BL/6 mice were randomly divided into 6 groups of 8 mice each group, including the control group, model group, RUT low-, medium-, and high-doses groups, and prednisone acetate group. The mice in each group were orally administered the corresponding drugs or vehicle once a day for 7 consecutive days. The gout inflammation model was established by intraperitoneal injection of MSU to evaluate the anti-gout inflammatory effects of RUT. Then the proinflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and the proportions of infiltrating neutrophils cytokines were detected by flow cytometry. In LPS/MSU-treated or untreated THP-1 macrophages, cell viability was observed by cell counting kit 8 and proinflammatory cytokines were measured by ELISA. The percentage of pyroptotic cells were detected by flow cytometry. Respectively, the mRNA and protein levels were measured by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot, the nuclear translocation of nuclear factor κB (NF-κB) p65 was observed by laser confocal imaging. Additionally, surface plasmon resonance (SPR) and molecular docking were applied to validate the binding ability of RUT components to tumor necrosis factor α (TNF-α) targets. RESULTS: RUT reduced the levels of infiltrating neutrophils and monocytes and decreased the levels of the proinflammatory cytokines interleukin 1β (IL-1β) and interleukin 6 (IL-6, all P<0.01). In vitro, RUT reduced the production of IL-1β, IL-6 and TNF-α. In addition, RT-PCR revealed the inhibitory effects of RUT on the mRNA levels of IL-1β, IL-6, cyclooxygenase-2 and TNF-α (P<0.05 or P<0.01). Mechanistically, RUT markedly reduced protein expressions of tumor necrosis factor receptor (TNFR), phospho-mitogen-activated protein kinase (p-MAPK), phospho-extracellular signal-regulated kinase, phospho-c-Jun N-terminal kinase, phospho-NF-κB, phospho-kinase α/β, NOD-like receptor thermal protein domain associated protein 3 (NLRPS), cleaved-cysteinyl aspartate specific proteinase-1 and cleaved-gasdermin D in macrophages (P<0.05 or P<0.01). Molecularly, SPR revealed that RUT bound to TNF-α with a calculated equilibrium dissociation constant of 31.7 µmol/L. Molecular docking further confirmed that RUT could interact directly with the TNF-α protein via hydrogen bonding, van der Waals interactions, and carbon-hydrogen bonding. CONCLUSION RUT alleviated MSU-induced peritonitis and inhibited the TNFR1-MAPK/NF-κB and NLRP3 inflammasome signaling pathway to attenuate gouty inflammation induced by LPS/MSU in THP-1 macrophages, suggesting that RUT could be a potential therapeutic candidate for gout.
Animals ; NF-kappa B/metabolism* ; Male ; Indole Alkaloids/therapeutic use* ; Signal Transduction/drug effects* ; Mice, Inbred C57BL ; Inflammation/complications* ; Uric Acid ; Quinazolines/therapeutic use* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Humans ; Gout/chemically induced* ; Inflammasomes/metabolism* ; Cytokines/metabolism* ; THP-1 Cells ; Mitogen-Activated Protein Kinases/metabolism* ; Mice ; Molecular Docking Simulation ; Lipopolysaccharides ; Quinazolinones

Objective To investigate the protective effect of verapamil on hyperuricemia nephropathy(HN)in mice through modulating TXNIP/NLRP3 inflammasome signaling pathway.Methods Thirty-two male C57BL/6J mice(8 weeks old,weighing 18～22 g)were randomly divided into a blank control group,a model group,an allopurinol group(10 mg/kg),and a verapamil group(40 mg/kg),with 8 animals in each group.Except for the control mice,the other mice were given 10%fructose water and adenine to establish a mouse model of HN.After successful establishment of model mice,the corresponding interventions were administered to the mice of the other 3 groups for 4 consecutive weeks.The levels of serum uric acid(UA),creatinine(Cr),urea(UREA),aspartate aminotransferase(AST)and alanine aminotransferase(ALT)were measured.HE staining was used to assess the alterations in renal morphology and the infiltration of inflammatory cells,while Masson's staining was employed to evaluate renal fibrosis.Moreover,ELISA was employed to measure the contents of IL-1β and IL-6 in kidney tissue,while serum levels of malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)were detected by colorimetric assay.Furthermore,immunohistochemical staining and Western blot analysis were conducted to examine the expression of TXNIP,NLRP3,IL-1β,MMP7,FN1,CD68,and MPO proteins in the kidney.Results Compared to the control group,HN mice exhibited increased serum UA,Cr,and UREA levels(P<0.05),renal pathological changes including renal tubular regeneration,interstitial or periglomerular fibrosis and prominent infiltration of inflammatory cells,and significantly increased renal contents of IL-1β and IL-6 and serum MDA level(P<0.05),while reduced serum SOD and GSH-Px contents(P<0.05),as well as up-regulation of kidney proteins TXNIP,NLRP3,IL-1β,CD68,MPO,FN1 and MMP7(P<0.01).Verapamil treatment notably reduced serum UA and Cr levels(P<0.01),improved kidney lesions to some extents,decreased collagen volume fraction(CVF)(P<0.01),and restored pro-inflammatory cytokines and oxidative stress markers(P<0.05)when compared with the levels in the model group.Further research found that the expression of kidney proteins TXNIP,NLRP3,IL-1β,CD68,MPO,FN1,and MMP7 was significantly down-regulated by verapamil treatment(P<0.05).Conclusion Verapamil exhibits a renal protective effect on HN mice through its anti-inflammatory,antioxidant,and antifibrotic properties,and its mechanism may be related to the inhibition of the TXNIP/NLRP3 inflammasome signaling pathway.

ObjectiveTo analyze the correlation between 11 small molecule active components and 1 protein component of characteristic processed products with porcine cardiac blood and other products of Salviae Miltiorrhizae Radix et Rhizoma（SMRR） from Menghe medical school and anti-cerebral ischemic oxidative damage， and to identify its key component markers of characteristic processed products with porcine cardiac blood for anti-cerebral ischemic oxidative damage. MethodHigh performance liquid chromatography（HPLC） was established to simultaneously determine the contents of 11 active ingredients in SMRR and its processed products［processed with porcine cardiac blood， porcine blood， wine and transferrin（Tf） in porcine cardiac blood］， and the content of Tf in different processed products of SMRR was detected by enzyme-linked immunosorbent assay（ELISA）. Furthermore， A zebrafish ischemic stroke model was constructed to evaluate the effects of different processed products of SMRR on the behavioral trajectory of cerebral ischemic zebrafish， the neuronal damage of transgenic zebrafish Tg（elavl3：eGFP） brain， as well as the levels of malondialdehyde（MDA） and superoxide dismutase（SOD） in the brain tissues. The hippocampal neurons oxygen-glucose deprivation/reoxygenation（OGD/R）-induced ischemia-hypoxia model was constructed to evaluate the effects of different processed products of SMRR on oxidative damage of neuronal cells by taking lactate dehydrogenase（LDH）， reactive oxygen species（ROS）， MDA and SOD as indexes. Finally， principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and Spearman correlation analysis were used to analyze the 11 small molecule active components and 1 protein component with efficacy indicators， in order to screen the key components of the characteristic processed products with porcine cardiac blood for cerebral ischemic oxidative damage. ResultCompared with the raw products， the contents of water-soluble and fat-soluble components in processed products of SMRR increased to different degrees， while the content of salvianolic acid A decreased. Compared with the wine-processed products， the contents of salvianolic acid B， danshensu， rosmarinic acid and other components in the porcine cardiac blood-processed products， porcine blood-processed products， Tf-processed products were increased， while the content of salvianolic acid A was decreased. ELISA results showed that there was no significant difference in Tf content between the porcine cardiac blood-processed products， porcine blood-processed products， Tf-processed products. Pharmacological results showed that different processed products of SMRR could improve the behavioral deficits， brain neuronal injury and oxidative stress after ischemic stroke in zebrafish， and the effect of the porcine cardiac blood-processed products was most pronounced. PCA results showed that salvianolic acid B， salvianolic acid A， rosmarinic acid， lithospermic acid， danshensu， tanshinone Ⅱ_A， caffeic acid， cryptotanshinone and tanshinone Ⅰ were the main contributing components of SMRR and its processed products. And the results of correlation analysis showed that the contents of cryptotanshinone， rosmarinic acid， caffeic acid， dihydrotanshinone Ⅰ， salvianolic acid B， tanshinone Ⅱ_A and tanshinone Ⅰ were negatively correlated with MDA level in zebrafish brain tissue， while the contents of lithospermic acid， protocatechuic aldehyde， rosmarinic acid， dihydrotanshinone Ⅰ， salvianolic acid B and Tf were positively correlated with SOD level， and the contents of rosmarinic acid， caffeic acid， dihydrotanshinone Ⅰ， salvianolic acid B， tanshinone Ⅱ_A， tanshinone Ⅰ， danshensu， Tf were positively correlated with neuronal fluorescence intensity in the zebrafish brain. And the contents of lithospermic acid， protocatechuic aldehyde， rosmarinic acid， dihydrotanshinone Ⅰ， salvianolic acid B， tanshinone Ⅱ_A and Tf were negatively correlated with LDH， ROS and MDA levels and positively correlated with SOD level. ConclusionThere are differences in the anti-ischemic oxidative damage effects of SMRR and its different processed products， among which the porcine cardiac blood-processed products has the strongest effect on improving oxidative damage， which may be related to the content changes of salvianolic acid B， danshensu， rosmarinic acid and other components. This study can provide a basis for clarifying the quality markers of SMRR processed with porcine cardiac blood for cerebral ischemia and elucidating its processing mechanism.

Objective To investigate the short-term efficacy of celiac plexus block combined with the interstitial permanent implantation of 125I seeds in the treatment of advanced pancreatic cancer(inoperable locally advanced and metastatic pancreatic cancers).Methods A total of 100 patients with advanced pancreatic cancer were selected and treated with celiac plexus block combined with the interstitial permanent implantation of 125I seeds under CT guidance.Pain relief and changes in the tumor marker CA19-9 were assessed on the seventh day,the first and third months after surgery.In the third month after surgery,tumor size was assessed by CT.Results Among the 100 patients with advanced pancreatic cancer,complete response(CR)was found in 12 cases,partial response(PR)in 78 cases,stable disease in five cases,and progression of disease in five cases three months after surgery.The CA19-9 level and the sum of short and long tumor diameters were significantly decreased(both P<0.01).A total of 100 patients had severe pain before treatment(visual analogue scale(VAS)):7-10 points),59 patients reported pain disappearance(VAS:0 points),35 patients had mild pain(VAS:1-3 points),and six patients experienced moderate pain(VAS:4-6 points)in the third month after treatment.The pain relief rate was 100%.Conclusion Celiac plexus block combined with the interstitial permanent implantation of 125I seeds has good short-term efficacy and can effectively improve short-term pain in patients with advanced pancreatic cancer.

Objective:To explore the feasibility and efficacy for the dissection and ligation of the superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer.Methods:Eight cadaveric heads were selected, and the laryngopharynxes were harvested. The positions of the superior laryngeal arteries entering the larynxes were dissected and observed under endoscopic vision, and their anatomical characteristics were summarized. Twenty-nine patients (all were male, aged 39-74 years old) with hypopharyngeal cancer who underwent transoral endoscopic surgery at the Department of Otorhinolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were selected, and the patients were randomly divided into two groups by drawing lots, namely, the superior laryngeal artery was actively dissected and occluded during surgery in observation group ( n=15) or not in control group ( n=14). The differences in surgical time, bleeding volume, postoperative complications, and postoperative disease-free survival rate were compared between the two groups. Statistical analysis was conducted using SPSS 25.0 software. Results:The entry point of the superior laryngeal artery into the larynx was approximately at the level of the superior edge of the thyroid cartilage, and entered the larynx at the posterior one-third of the lateral wall of the pyriform fossa. The superior laryngeal artery might be determined through endoscopic exploration in all patients of observation group. The endoscopic surgery time [(40.00±7.56) minutes] and intraoperative bleeding volume [(24.00±8.28) ml] in the observation group were respectively less than those [(48.57±14.06) minutes and (42.86±15.41) ml] in the control group, and the differences were statistically significant ( t=-2.064, P=0.049; t=-4.064, P=0.001). There was no case with postoperative bleeding in the observation group, but with one case of postoperative bleeding in the control group. Total disease free survival rate was 86.2% and there was no significant difference in disease free survival rates between the two groups during a follow-up period of at least 36 months ( P=0.986). Conclusion:Dissection of the superior laryngeal artery during endoscopic surgery for hypopharyngeal cancer is feasible, and pre-management and occlusion of the superior laryngeal artery can effectively reduce intraoperative bleeding.

Objective:To understand the breastfeeding situation in the neonatal intensive care units (NICUs) in Qianxinan Prefecture, Guizhou Province, and to assess the knowledge, attitudes, and practices of doctors and nurses regarding breastfeeding, aiming to provide foundational data for improving breastfeeding quality.Methods:A questionnaire was developed to survey the knowledge, attitudes, and practices related to breastfeeding in NICUs. The questionnaire was divided into three dimensions: knowledge (seven items, total score of 7), attitudes (nine items, total score of 45), and practices (seven items, total score of 35). Lower scores indicated weaker recognition of breastfeeding. Additionally, five items were included to identify the most influential factors affecting breastfeeding. From November 25 to November 30, 2023, a survey was conducted among doctors and nurses with professional qualifications who had worked in the neonatal departments of nine hospitals in Qianxinan Prefecture for at least one year. Independent sample t-tests and Chi-square tests were used to compare the scores of doctors and nurses from different levels of hospitals and within the same level of hospitals across the three dimensions. Results:(1) Among the nine hospitals, three were tertiary grade A hospitals (referred to as "tertiary hospitals"), with 95.6% (43/45) of the doctors and 96.5% (110/114) of the nurses participating in the survey. Six were secondary grade A hospitals (referred to as "secondary hospitals"), with 95.0% (38/40) of the doctors and 97.6% (83/85) of the nurses participating. (2) All nine hospitals were baby-friendly hospitals and all had breastfeeding promotional materials. Six hospitals had NICUs that promoted breastfeeding, with an average NICU breastfeeding rate of 25.8% across the prefecture between year 2021 to 2023. (3) The proportion of doctors who had received breastfeeding training was higher than that of nurses within the same level of hospitals [tertiary hospitals: 69.8% (30/43) vs. 40.0% (44/110), χ 2=10.97, P=0.001; secondary hospitals: 47.4% (18/38) vs. 24.1% (20/83), χ 2=6.55, P=0.010], although the overall training rates were low. (4) In tertiary hospitals, doctors scored higher than nurses in the attitude dimension [(35.35±4.75) vs. (33.18±5.60) scores, t=－2.03, P=0.044] and also in the practice dimension [(26.98±3.00) vs. (25.60±3.75) scores, t=－2.17, P=0.032]. In secondary hospitals, the total knowledge dimension score of doctors was higher than that of nurses [(4.92±1.44) vs. (4.20±1.45) scores, t=－2.52, P=0.013]. In tertiary hospitals, the total scores for attitude and practice dimensions of doctors were higher than those of doctors in secondary hospitals, and the total scores for knowledge, attitude, and practice dimensions of nurses were higher than those of nurses in secondary hospitals (all P<0.05). (5) In the knowledge dimension, the lowest scoring item of doctors in the tertiary hospitals was "Breastfeeding is possible for maternal hepatitis B newborns after receiving vaccines and immunoglobulin"; the lowest scoring item of nurses in the tertiary hospital, and doctors and nurses in the secondary hospitals was "The duration of breastfeeding has a greater impact on neonatal outcomes". In the attitude dimension, the lowest scoring item for doctors and nurses in both tertiary and secondary hospitals was "You think the breastfeeding process is more troublesome than feeding preterm formula". In the practice dimension, the lowest scoring item of the doctors and nurses in the tertiary hospitals was "Your hospital had enough breastfeeding knowledge training", while for the doctors and nurses in the secondary hospitals were "You have more opportunities to participate in various breastfeeding-related training" and "Breast feeding should be started as soon as possible when the infant is stable after active treatment", respectively. (6) The most influential factors affecting breastfeeding were: lack of cooperation from parents (50.0%, 137/274), relative insufficient human resources for doctors and nurses (21.9%, 60/274), and the absence or poor implementation of breastfeeding management policies (18.3%, 50/274), etc. Conclusions:The breastfeeding rate in NICU of county-level hospitals is relatively low, and medical staff, especially nurses, have insufficient knowledge about breastfeeding. It is necessary to strengthen various breastfeeding training for medical staff to enhance their understanding of NICU breastfeeding.

Objective:To compare the clinical efficacy of robot-assisted versus fluoroscopy-assisted sacroiliac screw internal fixation for posterior pelvic ring fractures.Methods:China National Knowledge Infrastructure (CNKI), Wanfang Data, Chinese Medical Journal Full-text Database, PubMed, Web of Science and ScienceDirect were searched for literature on robot-assisted versus fluoroscopy-assisted sacroiliac screw internal fixation for posterior pelvic ring fractures. The search time was from the establishment of each database to March 2023. Meta-analysis was performed on the included literature. The random-effects model was used when the heterogeneity between groups was large, and the fixed-effects model was used when the heterogeneity between groups was small.Results:A total of 15 studies were included in the meta-analysis, including 465 patients in the robot-assisted group and 396 patients in the fluoroscopy-assisted group. Meta-analysis showed that the number of fluoroscopies [ SMD=-3.12, 95% CI (-4.34, -1.89), P<0.001], the number of guide pin adjustments [ SMD=-3.75, 95% CI (-6.77, -0.72), P=0.015], intraoperative blood loss [ SMD=-0.83, 95% CI (-1.18, -0.49), P<0.001], and operative time [ SMD=-2.59, 95% CI (-4.11, -1.08), P<0.001] were smaller than those in the fluoroscopy-assisted group. The rate of excellent screw implantation [ OR=10.13, 95% CI (3.67,27.98), P<0.001] of the robot-assisted was larger than the fluoroscopy-assisted group. There was no significant difference in Majeed functional score [ SMD=0.28, 95% CI (-0.0003, 0.55), P=0.050] and fracture healing time [ SMD=-0.14, 95% CI (-0.46, 0.17), P=0.367] between the two groups. Conclusion:Robot-assisted percutaneous sacroiliac screw fixation for posterior pelvic ring fractures has the advantages of less fluoroscopy, less guide pin adjustment, less intraoperative blood loss, shorter operation time, and higher rate of excellent screw position. However, there is no difference in Majeed score and fracture healing time between robot-assisted percutaneous sacroiliac screw fixation and fluoroscopy-assisted percutaneous sacroiliac screw fixation.

OBJECTIVE: To explore the molecular mechanisms of Porphyromonas gingivalis infection-induced umbilical vein endothelial barrier dysfunction in vitro. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in vitro, and after the formation of the endothelial barrier, the cells were infected with P. gingivals at a multiplicity of infection (MOI). The transepithelial electrical resistance (TEER) of the cell barrier was measured, and FITC-dextran trans-endothelial permeability assay and bacterial translocation assay were performed to assess the endothelial barrier function. The expression levels of cell junction proteins including ZO-1, occludin and VE-cadherin in the cells were examined by qRT-PCR and Western blotting. RESULTS: In freshly seeded HUVECs, TEER increased until reaching the maximum on Day 5 (94 Ωcm²), suggesting the formation of the endothelial barrier. P. gingivals infection caused an increase of the permeability of the endothelial barrier as early as 0.5 h after bacterial inoculation, and the barrier function further exacerbated with time, as shown by significantly lowered TEER, increased permeability of FITC-dextran (40 000/70 000), and increased translocation of SYTO9-E. coli cross the barrier. MTT assay suggested that P. gingivals infection did not significantly affect the proliferation of HUVECs (P>0.05), but in P. gingivalsinfected cells, the expressions of ZO-1, occludin and VE-cadherin increased significantly at 24 and 48 h after bacterial inoculation (P < 0.05). CONCLUSION P. gingivals may disrupt the endothelial barrier function by down-regulating the expressions of the cell junction proteins (ZO-1, occludin, VE-cadherin) and increasing the permeability of the endothelial barrier.
Humans ; Cadherins/metabolism* ; Escherichia coli/metabolism* ; Human Umbilical Vein Endothelial Cells/metabolism* ; Occludin ; Porphyromonas gingivalis/metabolism* ; Umbilical Veins/metabolism*

To investigate the regulation of N6- methyladenosine ( m6A ) modification on L-type calcium channels in atrial myocytes under high hydrostatic pressure, mediated by methyltransferase-like protein 3 ( METTL3 ). Methods C57BL/6J mice were randomly assigned to the control group and the hypertension group ( treated with continuous administration of angiotensin for four weeks ). Masson staining was used to observe the fibrosis of mouse atrial tissue, while dot blot assay and Western blot were used to detect the levels of m6A, METTL3, and Cavi1 2 in the atrial tissue. A high hydrostatic pressure model was constructed using the HL-1 cell line cultured in vitro, and METTL3 was intervened to observe changes in m6A expression levels, METTL3 and Cavi1 2 levels in cells,and action potential duration ( APD ) and L-type calcium current ( I