Microglia, the resident immune cells in the central nervous system (CNS), rapidly transition from a resting to an active state in the acute phase of ischemic brain injury. This active state mediates a pro-inflammatory response that can exacerbate the injury. Targeting the pro-inflammatory response of microglia in the semi-dark band during this acute phase may effectively reduce brain injury. Shionone (SH), an active ingredient extracted from the dried roots and rhizomes of the genus Aster (Asteraceae), has been reported to regulate the inflammatory response of macrophages in sepsis-induced acute lung injury. However, its function in post-stroke neuroinflammation, particularly microglia-mediated neuroinflammation, remains uninvestigated. This study found that SH significantly inhibited lipopolysaccharide (LPS)-induced elevation of inflammatory cytokines, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS), in microglia in vitro. Furthermore, the results demonstrated that SH alleviated infarct volume and improved behavioral performance in middle cerebral artery occlusion (MCAO) mice, which may be attributed to the inhibition of the microglial inflammatory response induced by SH treatment. Mechanistically, SH potently inhibited the phosphorylation of serine-threonine protein kinase B (AKT), mammalian target of rapamycin (mTOR), and signal transducer and activator of transcription 3 (STAT3). These findings suggest that SH may be a potential therapeutic agent for relieving ischemic stroke (IS) by alleviating microglia-associated neuroinflammation.
Animals ; Microglia/immunology* ; Mice ; Male ; Mice, Inbred C57BL ; Brain Ischemia/immunology* ; Neuroinflammatory Diseases/drug therapy* ; Neuroprotective Agents/administration & dosage* ; Interleukin-1beta/genetics* ; STAT3 Transcription Factor/genetics* ; TOR Serine-Threonine Kinases/genetics* ; Tumor Necrosis Factor-alpha/genetics* ; Proto-Oncogene Proteins c-akt/immunology* ; Nitric Oxide Synthase Type II/genetics* ; Lipopolysaccharides

Objective:To investigate the precise advantages of robot-assisted total knee arthroplasty(RATKA)in reconstruction of mechanical axis of lower limb,and its influence on early functional rehabilitation.Methods:A prospective cohort design was used.A total of eighty patients with osteoarthritis who underwent total knee arthroplasty(TKA)at the Department of Joint Surgery of The People's Hospital of Suzhou New District between June 2021 and August 2024 were enrolled.According to different surgical method,they were divided into robot-assisted group(n=40)and conventional surgery group(n=40).The deviation angles of mechanical axis were quantified by three-dimensional(3D)imaging under weight-bearing conditions of 3D modeling(EOS)of whole body skeleton at the postoperative 1st week,the 3rd month,and the 6th month.The recovery of function was assessed by using the Hospital for Special Surgery(HSS)knee joint score.Results:The postoperatively average deviation angle of mechanical axis in the robot-assisted group was(1.23±0.67)°,which was significantly lower than the(2.89±1.23)° of the conventional surgery group(t=7.560,P<0.05).The increased extent of angular deviation was smaller with the change of time in the robot-assisted group.The HSS scores at the 6th month after surgery indicated that the score of relieving pain was(28.45±2.34)points in the robot-assisted group,and that was(25.12±3.45)points in the conventional surgery group,and the difference of that between two groups was significant(t=5.12,P<0.05).The score of joint stability was(9.23±0.78)points in the robot-assisted group,and that was(7.89±1.23)points in the conventional surgery group,and the difference of that between two groups was significant(t=5.89,P<0.05).The advantages of the robot-assisted group was significant.The operative time of the robot-assisted group was(98.45±10.23)min,which was significantly longer than the(85.67±9.78)min of the conventional surgery group(t=5.89,P<0.05).However,intraoperative blood loss in the robot-assisted group was(150.34±25.67)mL,which was significantly better than the(220.45±30.12)ml in the conventional surgery group(t=11.23,P<0.05).The total incidence of complication was 5.0%(2/40)in the robot-assisted group,and that was 17.5%(7/40)in the conventional surgery group,and the difference of that between two groups was significant(x2=4.01,P<0.05).Conclusion:RATKA can enhance significantly the reconstructed precision of lower limb alignment through 3D dynamic navigation.The characteristic of its sub-millimeter osteotomy can optimize early functional rehabilitation and reduce risk of postoperative complication.

Objective To investigate the biomechanical differences in plantar pressure,postural stability,and plantar Visual Analogue Scale(VAS)scores between normal feet and unilateral/bilat-eral pes cavus,reveal their unique neuromechanical compensation mechanisms,and construct a sta-bility early warning model based on the minimum center of pressure(COP)trajectory classification.Methods A total of 70 patients with pes cavus from December 2023 to October 2024 were selected as study subjects,including 33 patients in the unilateral pes cavus group and 37 patients in the bilat-eral pes cavus group.During the same period,32 normal feet were included as normal foot group.A flat-panel plantar pressure testing system was used to collect dynamic plantar pressure data and COP trajectories from three groups at a self-selected walking speed.There were no statistically significant differences in baseline data such as age,gender,and body mass index among the three groups(P＞0.05).One-way analysis of variance and the Wilcoxon rank-sum test were used to compare the differences in maximum pressure,contact area,VAS scores,and the 95％confidence ellipse area of the COP among the three groups in 10 plantar regions.Results Patients with pes cavus exhibited lower peak pressure in the MF region compared to normal feet,while higher peak pressure in the M2,M3,and MH regions.Patients with bilateral pes cavus showed lower peak pressure in the T1 region compared to normal feet,and patients with unilateral pes cavus had lower peak pressure in the LH region compared to the normal group(P＜0.05).The plantar contact area in patients with pes cavus was reduced in the T1,M2,M3,M4,MF,and MH regions compared to normal feet(P＜0.05).The 95％confidence ellipse area of the COP was larger in both the bilateral and uni-lateral pes cavus groups compared to the normal foot group(P＜0.001).Unilateral pes cavus pres-ented a specific lateral COP drift(amplitude of 3 to 4 cm),which is a biomechanical manifestation of compensatory eversion of the unaffected foot.Patients with bilateral pes cavus exhibited a"bimod-al oscillation"trajectory(amplitude of 6 to 8 cm),suggesting possible vestibular-spinal regulatory dysfunction and the poorest postural stability.In the pes cavus group,there was a significant in-crease in pressure in the M2,M3,and MH regions,with peak pressures exceeding 190 kPa in pa-tients with bilateral pes cavus,which was highly correlated with plantar pain and could serve as a pain early warning threshold.Conclusion Unilateral and bilateral pes cavus exhibit significantly different neuromechanical compensation patterns.The classification based on the"lateral drift"and"bimodal oscillation"characteristics of the COP trajectory can serve as a stability early warning indi-cator for assessing fall risk.Decompression interventions targeting the key pressure regions of M2,M3,and MH(such as customized orthotic insoles)are the core strategies for alleviating pain and optimizing dynamic gait stability.

Objective:To investigate the precise advantages of robot-assisted total knee arthroplasty(RATKA)in reconstruction of mechanical axis of lower limb,and its influence on early functional rehabilitation.Methods:A prospective cohort design was used.A total of eighty patients with osteoarthritis who underwent total knee arthroplasty(TKA)at the Department of Joint Surgery of The People's Hospital of Suzhou New District between June 2021 and August 2024 were enrolled.According to different surgical method,they were divided into robot-assisted group(n=40)and conventional surgery group(n=40).The deviation angles of mechanical axis were quantified by three-dimensional(3D)imaging under weight-bearing conditions of 3D modeling(EOS)of whole body skeleton at the postoperative 1st week,the 3rd month,and the 6th month.The recovery of function was assessed by using the Hospital for Special Surgery(HSS)knee joint score.Results:The postoperatively average deviation angle of mechanical axis in the robot-assisted group was(1.23±0.67)°,which was significantly lower than the(2.89±1.23)° of the conventional surgery group(t=7.560,P<0.05).The increased extent of angular deviation was smaller with the change of time in the robot-assisted group.The HSS scores at the 6th month after surgery indicated that the score of relieving pain was(28.45±2.34)points in the robot-assisted group,and that was(25.12±3.45)points in the conventional surgery group,and the difference of that between two groups was significant(t=5.12,P<0.05).The score of joint stability was(9.23±0.78)points in the robot-assisted group,and that was(7.89±1.23)points in the conventional surgery group,and the difference of that between two groups was significant(t=5.89,P<0.05).The advantages of the robot-assisted group was significant.The operative time of the robot-assisted group was(98.45±10.23)min,which was significantly longer than the(85.67±9.78)min of the conventional surgery group(t=5.89,P<0.05).However,intraoperative blood loss in the robot-assisted group was(150.34±25.67)mL,which was significantly better than the(220.45±30.12)ml in the conventional surgery group(t=11.23,P<0.05).The total incidence of complication was 5.0%(2/40)in the robot-assisted group,and that was 17.5%(7/40)in the conventional surgery group,and the difference of that between two groups was significant(x2=4.01,P<0.05).Conclusion:RATKA can enhance significantly the reconstructed precision of lower limb alignment through 3D dynamic navigation.The characteristic of its sub-millimeter osteotomy can optimize early functional rehabilitation and reduce risk of postoperative complication.

Objective:To assess the prognostic value of methylation of interferon regulatory factor 6 ( IRF6) gene promoter in patients diagnosed with Kidney renal clear cell carcinoma (KIRC). Methods:The primary lesions of fifty KIRC patients who were diagnosed at the First Affiliated Hospital of Nanjing Medical University from January 2016 to January 2020 were collected. The expression of IRF6 protein was determined with an immunohistochemical method. The correlation between the level of IRF6 expression and survival and/or metastasis status was analyzed. The mRNA and protein levels of the IRF6 in KIRC and normal renal tissues were compared by using bioinformatic tools. The difference in the methylation rate of the IRF6 gene promoter between tumor and adjacent tissues was analyzed by searching the online databases. Statistical analysis was carried out for the methylation status of the IRF6 gene promoter region to select those negatively correlated with the overall survival (OS) among the patients. In vitro experiments were conducted with cell lines to verify the correlation between the status of promoter methylation and transcription level of the IRF6 gene. Results:The mRNA and protein levels of the IRF6 gene in KIRC tissues were significantly lower than those of the normal controls, and this was more prominent in patients who had died or developed metastasis. The extent of IRF6 gene promoter methylation in the KIRC tissues was much higher compared with that of the adjacent normal renal tissues. There was a significant negative correlation between the methylation of the IRF6 gene promoter and mRNA level of the IRF6 ( R= -0.52). The higher methylation degree in the IRF6 gene promoter regions cg12034118 and cg16030177, the shorter the OS and worse prognosis in the patients. Only twenty CpG sites in cg12034118 were confirmed to be highly methylated in KIRC cell lines. The transcription level of the IRF6 gene was upregulated in a time- and dose-dependent manner after the treatment with demethylation reagent 5-azadeoxycytidine. Conclusion:The methylation of IRF6 gene promoter in the renal tissues of KIRC patients is closely correlated with the OS. Cg12034118 may provide a promising biomarker for laboratory detection, and its high methylation rate has certain reference value for the prognosis.

To assess the application value of white light endoscopic targeted biopsy in Tibetan gastric cancer risk population in Xizang region, a prospective study was conducted and a total of 666 Tibetan subjects were recruited who were assessed as medium or high risk by a novel gastric cancer screening scoring system at the Department of Gastroenterology, Lhasa People's Hospital from June 2022 to December 2023. Patients were randomly divided into the experimental group (339 cases, underwent white light endoscopic targeted biopsy) and the control group (327 cases, underwent white light endoscopic traditional biopsy) using a random number table method. In the experimental group, 254 (74.9%) cases underwent white light endoscopic targeted mucosal biopsies after evaluation of the targeted mucosal biopsy strategy. The detection rates of gastric mucosal atrophy [31.1% (79/254) VS 19.9% (65/327), χ2=9.66, P=0.002] and intestinal metaplasia [22.4% (57/254) VS 15.9% (52/327), χ2=4.01, P=0.045] were significantly higher in the experimental group compared with the control group. The detection rates of early gastric cancer [7.5% (19/254) VS 4.0% (13/327), χ2=3.37, P=0.066] and advanced gastric cancer [0.0% (0/254) VS 2.1% (7/327), P=0.617] were not significantly different between the two groups. The number of biopsy sites [3 (2, 4) sites VS 5 (5, 5) sites, Z=10.50, P<0.001], the number of biopsy attempts [4 (3, 5) times VS 5 (5, 5) times, Z=8.43, P<0.001], and the examination costs [1 444.05 (1 211.20, 1 775.98) yuan VS 1 529.47 (1 351.73, 1 777.48) yuan, Z=1.96, P=0.001] were significantly lower in the experimental group than those in the control group. Among the 85 subjects in the experimental group who did not undergo biopsy, 9 cases [2.7% (9/339)] developed mucosal atrophy and 13 cases [3.8% (13/339)] developed intestinal metaplasia during a 12-month follow-up period, with no case of early or advanced gastric cancer. These findings suggest that for the Tibetan population at gastric cancer risk in the Tibet region, white light endoscopic targeted biopsy can effectively increase the detection rates of gastric mucosal atrophy and intestinal metaplasia while reduce biopsy rates, without significantly increasing the missed diagnosis rates of early and advanced gastric cancer. This approach can be promoted for clinical application.

Objective This study was to develop a"Spleen Deficiency Certificate Scale for County Residents"and test its reliability.It was then developed as an objective tool for Chinese medicine evidence and symptoms for the prevention and control of chronic diseases among county residents.Methods The scale was compiled based on the team's previous foundation.The reliability of the scale was evaluated using internal consistency reliability and split-half reliability,while its validity was evaluated using structural validity,content validity,calibration validity,and discriminant validity.Results The study included 213 adults from Lanxi,of whom 155 were tested for intestinal flora.Seven scale entries were identified:Fatigue,fear of cold,bland mouth,loss of appetite,diarrhea,weak bowel movements,and tooth-marked tongue.In the reliability test,Cronbach's alpha coefficient was 0.828 and McDonald's ω coefficient was 0.825.The"stomach pain"and"bloating"entries did not meet the inclusion requirements and were recommended to be deleted.The Spearman-Brown coefficient was 0.839.The exploratory factor analysis of the two common factors explained 61.6%of the cumulative variance.The calibration validity indicated that the ratio of salivary amylase activity before and after acid stimulation was 0.826±0.253 in the group with spleen deficiency.Significant differences(P<0.05)in the genera Dialister,Shigella,Leuconostoc,Photobacterium,Trabulsiella,and Parvimonas between the spleen deficiency group and the non-spleen deficiency group.Conclusion The Spleen Deficiency Scale for County Residents demonstrates good reliability and validity.

To assess the application value of white light endoscopic targeted biopsy in Tibetan gastric cancer risk population in Xizang region, a prospective study was conducted and a total of 666 Tibetan subjects were recruited who were assessed as medium or high risk by a novel gastric cancer screening scoring system at the Department of Gastroenterology, Lhasa People's Hospital from June 2022 to December 2023. Patients were randomly divided into the experimental group (339 cases, underwent white light endoscopic targeted biopsy) and the control group (327 cases, underwent white light endoscopic traditional biopsy) using a random number table method. In the experimental group, 254 (74.9%) cases underwent white light endoscopic targeted mucosal biopsies after evaluation of the targeted mucosal biopsy strategy. The detection rates of gastric mucosal atrophy [31.1% (79/254) VS 19.9% (65/327), χ2=9.66, P=0.002] and intestinal metaplasia [22.4% (57/254) VS 15.9% (52/327), χ2=4.01, P=0.045] were significantly higher in the experimental group compared with the control group. The detection rates of early gastric cancer [7.5% (19/254) VS 4.0% (13/327), χ2=3.37, P=0.066] and advanced gastric cancer [0.0% (0/254) VS 2.1% (7/327), P=0.617] were not significantly different between the two groups. The number of biopsy sites [3 (2, 4) sites VS 5 (5, 5) sites, Z=10.50, P<0.001], the number of biopsy attempts [4 (3, 5) times VS 5 (5, 5) times, Z=8.43, P<0.001], and the examination costs [1 444.05 (1 211.20, 1 775.98) yuan VS 1 529.47 (1 351.73, 1 777.48) yuan, Z=1.96, P=0.001] were significantly lower in the experimental group than those in the control group. Among the 85 subjects in the experimental group who did not undergo biopsy, 9 cases [2.7% (9/339)] developed mucosal atrophy and 13 cases [3.8% (13/339)] developed intestinal metaplasia during a 12-month follow-up period, with no case of early or advanced gastric cancer. These findings suggest that for the Tibetan population at gastric cancer risk in the Tibet region, white light endoscopic targeted biopsy can effectively increase the detection rates of gastric mucosal atrophy and intestinal metaplasia while reduce biopsy rates, without significantly increasing the missed diagnosis rates of early and advanced gastric cancer. This approach can be promoted for clinical application.

Immunotherapy has revolutionized the landscape of cancer treatment. However, single immunotherapy only works well in a small subset of patients. Combined immunotherapy with antitumor synergism holds considerable potential to boost the therapeutic outcome. Nevertheless, the synergistic, additive or antagonistic antitumor effects of combined immunotherapies have been rarely explored. Herein, we established a novel combined cancer treatment modality by synergizing p21-activated kinase 4 (PAK4) silencing with immunogenic phototherapy in engineered extracellular vesicles (EVs) that were fabricated by coating M1 macrophage-derived EVs on the surface of the nano-complex cores assembled with siRNA against PAK4 and a photoactivatable polyethyleneimine. The engineered EVs induced potent PAK4 silencing and robust immunogenic phototherapy, thus contributing to effective antitumor effects in vitro and in vivo. Moreover, the antitumor synergism of the combined treatment was quantitatively determined by the CompuSyn method. The combination index (CI) and isobologram results confirmed that there was an antitumor synergism for the combined treatment. Furthermore, the dose reduction index (DRI) showed favorable dose reduction, revealing lower toxicity and higher biocompatibility of the engineered EVs. Collectively, the study presents a synergistically potentiated cancer treatment modality by combining PAK4 silencing with immunogenic phototherapy in engineered EVs, which is promising for boosting the therapeutic outcome of cancer immunotherapy.