BACKGROUND:Currently,treatment method for knee osteoarthritis includes oral medicine,joint cavity drug injection,and physiotherapy,but the curative effect is limited.Existing studies have confirmed that silicon-based bioceramics can promote cartilage and subchondral bone repair and vascular regeneration.OBJECTIVE:To explore the effect of different concentrations of silicon-based bioceramics injected into the knee joint cavity in the treatment of knee osteoarthritis in rats.METHODS:Silicon-based bioceramics-calcium silicate was prepared.Twenty-five SD rats were randomly divided into five groups,with five rats in each group.The healthy group did not receive any intervention,and the modeling group,low-dose calcium silicate group,high-dose calcium silicate group,and saline group used anterior cruciate ligament transection to establish bilateral knee osteoarthritis models.Four weeks after modeling,0.05 mL of 50 and 100 mg/mL calcium silicate solution were injected into the knee joint cavity in the low-dose calcium silicate group and high-dose calcium silicate group,respectively,and 0.05 mL of saline was injected into the knee joint cavity in the saline group,once a week for 4 consecutive weeks.In the fifth week of administration,bilateral knee joint Micro-CT detection,knee joint cartilage hematoxylin-eosin staining,and modified Mankin score were performed.RESULTS AND CONCLUSION:(1)Micro-CT quantitative analysis showed that compared with the healthy group,the volume fraction and number of trabeculae of the medial tibial plateau in the modeling group decreased(P＜0.05),and the separation of trabeculae increased(P＜0.05).Compared with the modeling group,the volume fraction and number of trabeculae of the medial tibial plateau in the low-dose calcium silicate group and the saline group increased(P＜0.05),and the separation of trabeculae decreased(P＜0.05).(2)Hematoxylin-eosin staining showed that the cartilage surface of the healthy group and the low-dose calcium silicate group was relatively smooth and flat,the chondrocytes were evenly distributed,without clustered chondrocytes,the tide line was complete,and the staining was uniform;the cartilage surface of the high-dose calcium silicate group was slightly uneven,the middle and deep cells were disordered,with a small number of clustered chondrocytes,the tide line was discontinuous,and the staining was uneven;the cartilage surface of the saline group and the modeling group was obviously rough,the cells were disordered,with a large number of clustered chondrocytes,the tide line disappeared,and the staining was uneven.The modified Mankin score of the healthy group was lower than that of the high-dose calcium silicate group,the saline group,and the modeling group(P＜0.05).The modified Mankin score of the high-dose calcium silicate group and the low-dose calcium silicate group was lower than that of the saline group and the modeling group(P＜0.05).(3)The results show that calcium silicate knee joint injection has a certain effect in the treatment of knee osteoarthritis.Compared with 100 mg/mL calcium silicate solution,50 mg/mL calcium silicate solution can promote the recovery of subchondral bone and cartilage.

BACKGROUND:Currently,treatment method for knee osteoarthritis includes oral medicine,joint cavity drug injection,and physiotherapy,but the curative effect is limited.Existing studies have confirmed that silicon-based bioceramics can promote cartilage and subchondral bone repair and vascular regeneration.OBJECTIVE:To explore the effect of different concentrations of silicon-based bioceramics injected into the knee joint cavity in the treatment of knee osteoarthritis in rats.METHODS:Silicon-based bioceramics-calcium silicate was prepared.Twenty-five SD rats were randomly divided into five groups,with five rats in each group.The healthy group did not receive any intervention,and the modeling group,low-dose calcium silicate group,high-dose calcium silicate group,and saline group used anterior cruciate ligament transection to establish bilateral knee osteoarthritis models.Four weeks after modeling,0.05 mL of 50 and 100 mg/mL calcium silicate solution were injected into the knee joint cavity in the low-dose calcium silicate group and high-dose calcium silicate group,respectively,and 0.05 mL of saline was injected into the knee joint cavity in the saline group,once a week for 4 consecutive weeks.In the fifth week of administration,bilateral knee joint Micro-CT detection,knee joint cartilage hematoxylin-eosin staining,and modified Mankin score were performed.RESULTS AND CONCLUSION:(1)Micro-CT quantitative analysis showed that compared with the healthy group,the volume fraction and number of trabeculae of the medial tibial plateau in the modeling group decreased(P＜0.05),and the separation of trabeculae increased(P＜0.05).Compared with the modeling group,the volume fraction and number of trabeculae of the medial tibial plateau in the low-dose calcium silicate group and the saline group increased(P＜0.05),and the separation of trabeculae decreased(P＜0.05).(2)Hematoxylin-eosin staining showed that the cartilage surface of the healthy group and the low-dose calcium silicate group was relatively smooth and flat,the chondrocytes were evenly distributed,without clustered chondrocytes,the tide line was complete,and the staining was uniform;the cartilage surface of the high-dose calcium silicate group was slightly uneven,the middle and deep cells were disordered,with a small number of clustered chondrocytes,the tide line was discontinuous,and the staining was uneven;the cartilage surface of the saline group and the modeling group was obviously rough,the cells were disordered,with a large number of clustered chondrocytes,the tide line disappeared,and the staining was uneven.The modified Mankin score of the healthy group was lower than that of the high-dose calcium silicate group,the saline group,and the modeling group(P＜0.05).The modified Mankin score of the high-dose calcium silicate group and the low-dose calcium silicate group was lower than that of the saline group and the modeling group(P＜0.05).(3)The results show that calcium silicate knee joint injection has a certain effect in the treatment of knee osteoarthritis.Compared with 100 mg/mL calcium silicate solution,50 mg/mL calcium silicate solution can promote the recovery of subchondral bone and cartilage.

Health economics evidence plays an important role in linking clinical value evidence with health resource allocation decisions in the development of clinical practice guidelines. It can not only effectively balance clinical effectiveness and economic feasibility but also avoid forming "idealized" recommendations that are detached from the affordability of the healthcare system or the burden-bearing capacity of patients. To promote guideline developers to use health economics evidence more standardizedly and fully, this paper conducts an in-depth analysis of the current application status, existing challenges, access channels, and application processes of health economics evidence in current guidelines, and on this basis, puts forward considerations and suggestions for strengthening and standardizing the application of health economics evidence in China's clinical practice guidelines.

Viral pneumonia in the Lancang-Mekong River Basin exhibits distinct regional epidemiological characteristics and pathogenic mechanisms. Filthy dampness constitutes the core pathogenic factor of the disease here， and the core pathogenesis lies in the yang failing to be stored in the you （酉） position. Clinically， the fundamental therapeutic principle is established as boosting qi to expel filth and guiding yang qi to descend and converge into the you position. In the early stage， the pathogenesis is characterized by the pervasive diffusion of filthy dampness， yang qi floating externally and failing to be stored， and the lung failing to diffuse qi. The treatment focuses on invigorating yang qi to dispel filth and dredging the you pathways， primarily using modified Ganlu Xiaodu Decoction （甘露消毒丹）. In the progressive stage， the pathogenesis involves the tenacious stagnation of filthy dampness， yang constraint transforming into heat， and the rampant spread of heat-toxins. The therapeutic approach shifts to acrid medicinals for opening and bitter medicinals for promoting descent， rectifying healthy qi to expel filth， utilizing modified Dayuan Beverage （达原饮）. In the recovery stage， filthy dampness gradually recedes， accompanied by the depletion of qi and yin， and insufficient kidney yang. The treatment emphasizes supplementing qi and nourishing yin， accompanied by warming yang， with modified Yunv Decoction （玉女煎）.

Objective To establish a cell bank of extrahepatic cholangiocarcinoma (ECC)-derived organoids and investigate the key factors influencing the organoids generation. Methods The tumor samples from patients with portal cholangiocarcinoma (pCCA) and distal cholangiocarcinoma (dCCA) were used to isolate cells, and these cells were cultured using three-dimensional (3D) technique to establish ECC organoids. Histological characteristics of the organoids were evaluated and identified through hematoxylin-eosin (HE) and immunohistochemistry stainings. The success rates of organoids generation from different tumor types were compared. And clinical characteristics of patients between successful and failure culture groups were compared. Results The success rates of organoids establishment from pCCA and dCCA were all low, with 42.4% (14/33), 51.9% (14/27), respectively. The tumor was larger in successful group than that in failure group (P＜0.001); there was no statistical difference in tumor differentiation status, microvascular invasion, and perineural invasion between the two groups. Conclusions The successful rate of ECC-derived organoids establishment is low, and larger tumor has higher successful culture rate.

Objective:To investigate the correlation between the density and volume of left atrial-epicardial adipose tissue(LA-EAT)and the occurrence and severity of atrial fibrillation(AF)in elderly patients.Methods:This was a retrospective case-control study.A total of 197 elderly patients with AF and 82 patients with sinus rhythm who were hospitalized in the Department of Cardiology and Geriatric Cardiology of the First Affiliated Hospital of Nanjing Medical University from June 2023 to July 2024.were selected.The AF group was further divided into paroxysmal AF(PAF)subgroup(111 cases)and persistent AF(PeAF)subgroup(86 cases). The density and volume of LA-EAT in all patients were meansured by coronary artery enhanced CT or left atrial enhanced CT.The differences in LA-EAT density, volume and related data among the 3 groups were compared, and their relationship with different severity of senile AF was analyzed.Logistic regression was used to analyze whether LA-EAT density and volume were risk factors for senile AF.The predictive value of LA-EAT density and volume for AF in the elderly was compared throuh restricted cubic spline analysis.Results:The average LA-EAT density in the sinus rhythm group, PAF group and PeAF group was -82.75(-87.15, -79.63), -81.70(-85.55, -75.85)and -80.45(-83.40, -76.10)HU, respectively; the standard deviation of density was 34.70(31.93, 36.28), 36.20(34.30, 37.65)and 35.45(33.63, 37.28)HU, respectively; the LA-EAT volume was 23 483.40(13 964.10, 31 645.53), 25 112.20(18 479.10, 36 917.50)and 37 836.50(25 933.80, 45 537.90)mm 3, respectively.The LA-EAT density and volume in the PAF group and PeAF group were higher than those in the sinus rhythm group(all P<0.05). The LA-EAT volume in the PeAF group was greater than that in the PAF group( P<0.001), but there was no significant difference in LA-EAT density between the two groups( P>0.05). Multivariate logistic regression analysis showed that LA-EAT density and volume, left atrial diameter(LAD)and body mass index(BMI)were the independent risk factors for AF in the elderly( P<0.05). Conclusions:LA-EAT density and volume are significantly correlated with occurrence of senile AF, and show a positive dose-response relationship.LA-EAT volume is significantly related to the severity of AF( P<0.05), while the density is not significantly related to it( P>0.05).

To develop a guideline terminology system and promote its standardization, thereby enhancing medical staff's accurate understanding and correct application of guidelines.

Objective:To explore the role of retinoic acid-related orphan receptor α (RORα) in cognitive impairment induced by chronic alcohol consumption in mice.Methods:(1)The SPF grade RORαflox/flox transgenic mice aged 8 weeks were generated, and 22 transgenic mice were evenly divided into two groups by the method of matching body mass, which were the control group (Con group) and the alcohol group (EtOH group), with 11 mice in each group.(2)Emx1-Cre transgenic mice were used to selectively knock out the RORα gene in the forebrain of RORαflox/flox transgenic mice, producing conditional knockout mice (cKO mice) with the genotype RORαflox/flox-Emx1-Cre+ /+. Fourteen cKO mice were further split into two groups by the method of matching body mass, which were the conditional knockout group (cKO group) and the conditional knockout + alcohol group (cKO + EtOH group), with 7 mice in each group. A chronic alcohol use cognitive impairment model was developed in the EtOH group and cKO + EtOH group through the two-bottle free-choice method, while the Con group and cKO group were given two bottles of water for the same period. Cognitive abilities of mice in all groups were evaluated using behavioral novel object recognition test and Y-maze test.RORα mRNA and protein expression levels in the hippocampus of the Con group and EtOH group were assessed by RT-qPCR and Western blot, respectively.The GraphPad Prism 9.0 software was used for data analysis.One-way ANOVA was used for the comparison of multiple groups and Tukey test was used for further pairwise comparisons.Results:(1) Comparison between Con group and EtOH group: the relative levels of ROR α protein (0.63±0.04) and mRNA (0.78±0.03) in the hippocampus of mice in the EtOH group were significantly lower than those in the Con group((1.00±0.06), (1.00±0.05), both P<0.05). The duration of the EtOH group in the Y maze was significantly lower than that of the Con group ((212.30±32.05) s, (129.30±21.50) s, P<0.05), and the new object recognition index of the EtOH group was lower than that of the Con group ((14.73±25.49)% vs (-15.55±27.88)%, P=0.08). (2)Comparison between Con group and cKO group: the frequency and duration of entering the Y maze of mice in the cKO group ((7.43±2.30) times, (124.10±13.95) s) were lower than those in the Con group ((14.90±3.65) times, (212.30±32.05) s, both P<0.05). There was no statistically significant difference in the new object recognition index between the cKO group and the Con group ( P>0.05). (3) Comparison between the cKO+ EtOH group and the cKO group: the frequency ((2.71±1.11)times) and duration ((161.70±17.95) s) of entering the new heteroarm of Y maze in the KO+ EtOH group were lower than those in the cKO group ((7.43±2.30) times, (124.10±13.95) s, both P<0.05), and there was no statistically significant difference in the new object recognition index ( P>0.05). (4) Comparison between the cKO+ EtOH group and the EtOH group: the frequency of entering new heteroarm of the Y maze in the cKO+ EtOH group ((2.71±1.11)times) was significantly lower than that in the EtOH group (12.18±4.49) ( P<0.05), while there was no statistically significant difference in other behavioral results between the two groups ( P>0.05). Conclusion:Chronic alcohol consumption leads to cognitive impairment through the downregulation of RORα in the hippocampus of mice. Specific knockout of RORα in the forebrain exacerbates cognitive impairment induced by chronic alcohol use. RORα may represent a potential therapeutic target for cognitive impairment resulting from chronic alcohol consumption.

Objective:To investigate the effect of circular RNA bromodomain PHD finger transcription factor (circBPTF) targeting microRNA (miR)-224-3p on the damage of human retinal vascular endothelial cells (HRECs) induced by high glucose.Methods:HRECs were divided into control group and high glucose group, which were cultured with medium containing 5.5 mmol/L glucose and 30 mmol/L glucose for 48 hours, respectively.HRECs were transfected with siRNA negative control (si-NC), siRNA of circBPTF (si-circBPTF), miRNA mimic control (miR-NC), and miR-224-3p by Lipofectamine 2000, followed by cultured in 30 mmol/L glucose medium for 48 hours and were recorded as si-NC group, si-circBPTF group, miR-NC group and miR-224-3p group, respectively.HRECs were transfected with si-circBPTF and anti-miR-NC or si-circBPTF and anti-miR-224-3p by double transfection method, and then treated with 30 mmol/L glucose medium for 48 hours and were recorded as anti-miR-NC group and anti-miR-224-3p group, respectively.The expression of circBPTF and miR-224-3p was detected by real-time fluorescence quantitative PCR.Cell apoptosis was detected by flow cytometry.Reactive oxygen species (ROS) level was determined by 2′, 7′-dichlorodihydrofluorescein diacetate probe.Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected by colorimetric method.Expression of cleaved-caspase-3/caspase-3 and cleaved-caspase-9/caspase-9 proteins was detected by Western blot.The interaction between circBPTF and miR-224-3p was identified by the dual luciferase reporter method.Results:Compared with the control group, the apoptosis rate, cleaved-caspase-3/caspase-3 protein expression, cleaved-caspase-9/caspase-9 protein expression, ROS level, MDA content, and circBPTF relative expression were significantly increased and SOD activity and miR-224-3p expression were decreased in the high glucose group (all P<0.05).Compared with the si-NC group, circBPTF relative expression, ROS level, MDA content, cleaved-caspase-3/caspase-3 protein expression, cleaved-caspase-9/caspase-9 protein expression, and cell apoptosis rate were significantly reduced and SOD activity was significantly increased in the si-circBPTF group (all P<0.05).Compared with the miR-NC group, miR-224-3p relative expression and SOD activity were significantly enhanced and ROS level, MDA content, cleaved-caspase-3/caspase-3 protein expression, cleaved-caspase-9/caspase-9 protein expression and cell apoptosis rate were significantly reduced in the miR-224-3p group (all P<0.05).The relative luciferase activity of HRECs after co-transfection of wild type circBPTF and miR-224-3p mimic was 0.43±0.04, which was significantly decreased compared with 0.99±0.06 after co-transfection of wild type circBPTF and miR-NC ( t=23.297, P<0.05).Compared with anti-miR-NC group, the relative expression of miR-224-3p and SOD activity were significantly decreased and ROS level, MDA content, cell apoptosis rate, cleaved-caspase-3/caspase-3 protein expression, and cleaved-caspase-9/caspase-9 protein expression were significantly increased in the anti-miR-224-3p group (all P<0.05). Conclusions:Interfering with circBPTF can inhibit high glucose induced HRECs apoptosis and oxidative stress damage by targeting miR-224-3p.

Objective:To investigate the expressions of serum cytokines in patients with autoimmune retinopathy (AIR), and their association with disease diagnosis as well as clinical features.Methods:A prospective case-control study was conducted.A total of 90 eyes of 45 AIR patients were consecutively collected in Beijing Tongren Hospital from September 2018 to December 2023.Additionally, age-matched 43 controls (86 eyes) were enrolled, consisting of 21 patients (42 eyes) with retinitis pigmentosa as a disease control group and 22 healthy subjects (44 eyes) as a normal control group.The main clinical outcome measures included best-corrected visual acuity (BCVA), mean deviation (MD) of visual field, central retinal thickness and the maximal response amplitude and implicit time of full-field electroretinography (ff-ERG). A total of 21 serum cytokines were detected by Luminex multiple cytokines assay or ELISA.Differences in serum cytokine concentrations of among groups, the correlation between cytokine levels and clinical outcomes, and the contribution of cytokines to the diagnosis of AIR were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital (No.TRECKY2018-048). Written informed consent was obtained from each subject.Results:In AIR patients, the expression levels of Th1-type cytokines or receptors (including interferon-γ[IFN-γ], interleukin [IL]-8, C-X-C motif chemokine ligand [CXCL]9, and CXCL10) and Th17-type cytokines (including IL-17 and IL-6) were elevated.There were statistically significant overall differences in pro-inflammatory cytokines/chemokines (IL-6, IL-8, CXCL10, IL-10, IFN-γ, IL-17, C-X3-C motif chemokine ligand 1 [CX3CL]1, CXCL9) among the three groups ( H=10.823, 10.816, 9.633, 10.103, 23.670, 16.493, 9.050, 9.253; all P<0.05). Compared with the disease control group and the healthy control group, the IFN-γ level was significantly increased in the AIR group (both P=0.001). Compared with the healthy control group, the serum levels of IL-6, IL-8, CXCL10, IL-10, IL-17, and CXCL9 in the AIR group were significantly elevated (all P<0.05). The serum level of CX3CL1 was significantly higher in the AIR group than in the disease control group ( P=0.039). The serum level of IFN-γ was significantly higher in the AIR group than in the healthy control and disease control groups, but no association with AIR diagnosis was found ( OR=1.402, 95% CI: 0.710-2.870, P=0.245). Multilevel mixed-effects regression modeling analysis showed that the serum levels of IL-8 and CXCL9 in AIR patients were positively correlated with LogMAR BCVA ( β=0.028, P=0.033; β=0.023, P=0.003). The C-C motif chemokine ligand 11 level was positively correlated with implicit time of ERG a-wave ( β=13.950, P<0.001). The tumor necrosis factor-α level was positively correlated with MD value of visual field ( β=6.310, P=0.002). Granulocyte-macrophage colony-stimulating factor was negatively correlated with the amplitude of ERG a-wave and granulocyte colony-stimulating factor was negatively correlated with ERG b-wave amplitude ( β=-152.700, P<0.001; β=-14.790, P=0.003). Conclusions:Serum Th1 and Th17-type cytokines/chemokines may be involved in the pathogenesis of AIR and are related to disease severity.