Background: and Purpose Bilirubin has potent antioxidant, anti-inflammatory, and neuroprotective effects. Herein, we investigated whether total bilirubin (TBIL) modifies the association between hypertension and cerebral small vessel disease (CSVD). Methods: Data were obtained from the PolyvasculaR Evaluation for Cognitive Impairment and vaScular Events study. TBIL and direct bilirubin (DBIL) levels were assayed using fasting venous blood samples. Indirect bilirubin (IBIL) was calculated by subtracting DBIL from TBIL. TBIL was stratified as ≤17 μmol/L and >17 μmol/L based on the biological relevance of Gilbert’s syndrome. Hypertension was defined as blood pressure ≥140/90 mm Hg, self-reported hypertension history, or current use of antihypertensive agents. White matter hyperintensity, lacunes, cerebral microbleeds, and enlarged perivascular spaces were evaluated using magnetic resonance imaging and used to rate CSVD burden according to the criteria proposed by Wardlaw et al. and Rothwell et al. Results: This study included 3,061 participants, with a mean age of 61.2±6.7 years and 46.5% males. After adjusting for confounders, hypertension was associated with increased odds of presence of CSVD (Wardlaw: odds ratio [OR]=1.86, 95% confidence interval [CI] 1.41–2.44, P<0.001; Rothwell: OR=1.84, 95% CI 1.43–2.38, P<0.001) and higher modified total CSVD burden (common OR: 1.85, 95% CI 1.45–2.36, P<0.001) in participants with TBIL ≤17 μmol/L but not in TBIL >17 μmol/L (P for interaction <0.05). Johnson–Neyman analyses showed cut-off concentrations of 22.3–22.4 μmol/L for effect modification by TBIL. IBIL contributed to effect modification, whereas DBIL did not. Conclusions Mildly elevated TBIL may modify the association between hypertension and CSVD.

Background: and Purpose This study aimed to investigate the association between residual inflammatory risk (RIR) and vulnerable plaques using high-resolution magnetic resonance imaging (HRMRI) in symptomatic intracranial atherosclerotic stenosis (ICAS). Methods: This retrospective study included 70%–99% symptomatic ICAS patients hospitalized from January 2016 to December 2022. Patients were classified into four groups based on high-sensitivity C-reactive protein (hs-CRP) and low-density lipoprotein cholesterol (LDL-C): residual cholesterol inflammatory risk (RCIR, hs-CRP ≥3 mg/L and LDL-C ≥2.6 mmol/L), RIR (hs-CRP ≥3 mg/L and LDL-C <2.6 mmol/L), residual cholesterol risk (RCR, hs-CRP <3 mg/L and LDL-C ≥2.6 mmol/L), and no residual risk (NRR, hs-CRP <3 mg/L and LDL-C <2.6 mmol/L). Vulnerable plaque features on HRMRI included positive remodeling, diffuse distribution, intraplaque hemorrhage, and strong enhancement. Results: Among 336 included patients, 21, 60, 58, and 197 were assigned to the RCIR, RIR, RCR, and NRR groups, respectively. Patients with RCIR (adjusted odds ratio [aOR], 3.606; 95% confidence interval [CI], 1.346–9.662; P=0.011) and RIR (aOR, 3.361; 95% CI, 1.774–6.368, P<0.001) had higher risks of strong enhancement than those with NRR. Additionally, patients with RCIR (aOR, 2.965; 95% CI, 1.060–8.297; P=0.038) were more likely to have intraplaque hemorrhage compared with those with NRR. In the sensitivity analysis, RCR (aOR, 2.595; 95% CI, 1.201–5.608; P=0.015) exhibited an additional correlation with an increased risk of intraplaque hemorrhage. Conclusion In patients with symptomatic ICAS, RIR is associated with a higher risk of intraplaque hemorrhage and strong enhancement, indicating an increased vulnerability to atherosclerotic plaques.

Objective To explore the efficacy of precise surface positioning method in percutaneous screw fixation for Jones fracture.Methods Data of 46 patients with Jones fractures admitted to our hospital from September 2021 to December 2023 were retrospectively analyzed.According to the different treatment methods,the patients were divided into screw groups(24 patients fixed by percutaneous screws)and tension band groups(22 patients fixed by incision and reset tension band).The general conditions,follow-up time,surgical time,hospitalization time,fracture reduction quality,postoperative complete weight-bearing time,fracture healing time,and complications in the two groups were recorded and compared.The American Association of Foot and Ankle Surgery(AOFAS)system was used to assess the recovery of foot function.Results There was no statistically significant difference in the general data before the operation between the two groups of patients(P＞0.05).The follow-up time of the two groups was 6 to 24 months.Compared with the tension band group,the screw group had shorter operation time[(32.96±5.35)min vs.(47.27±9.42)min],shorter hospitalization time[(5.75±1.18)d vs.(7.68±2.05)d],earlier postoperative complete weight-bearing time[(6.47±0.73)w vs.(7.29±1.66)w],faster fracture healing time[(3.31±0.68)months vs.(4.29±1.29)months],higher AOFAS score[(89.13±2.62)vs.(79.59±4.49)](all P＜0.05).The tension band group of fracture reduction quality was better than the screw group(P＜0.05).The quality of fracture reduction during the operation in the tension band group was better than that in the screw group(P＜0.05).In terms of complications:no complications occurred in the screw group,one patient in the tension band group developed infection,and one patient had a loose Kirchner needle.Conclusion Precise surface positioning method in percutaneous screw fixation for Jones fracture is a safe and effective method with precise positioning,simple operation,small trauma,and rapid rehabilitation,which is worthy of clinical promotion,but the indications need to be strictly controlled according to the type of fracture.

Flow cytometer plays an indispensable role in the research of each field of life science and clinical medicine.This type of equipment has the characteristics of precision,high cost,precise operation,and high maintenance costs.Therefore,high requirements for technique and environmental are required for operators.Taking BD FACS Aria III flow cytometer as an example,the factors of working environment,the precautions of daily maintenance and care were analyzed.The common fault analysis and processing methods in start-up and shutdown for flow,instrument calibration,cell sorting,data acquisition and image display were described,so as to provide reference for the users of the similar equipment,and maximize the effectiveness of the instrument,and ensure the usage effect of equipment and the lifespan of instrument.

BACKGROUND:The blood-brain barrier is an important structure that protects the central nervous system,and the study of the effects of high glucose on the blood-brain barrier is important for the prevention of high glucose-induced damage to the central nervous system.OBJECTIVE:To investigate the potential effect of high glucose on the blood-brain barrier function of hCMEC/D3 human brain microvascular endothelial cells.METHODS:hCMEC/D3 cells were cultured in regular sugar medium(glucose concentration of 25 mmol/L)and high-sugar medium(glucose concentration of 55 mmol/L).The morphology of cells in each group was observed by light microscopy.CCK-8 assay was used to detect changes in cell viability.A monolayer blood-brain barrier model was established using hCMEC/D3 cell line with Transwell chamber device.Changes in cell transmembrane resistance were monitored daily.The permeability of cell monolayers was detected by phenol red permeability.Flow cytometry was used to detect the apoptosis rate of the cells.Western blot assay was used to detect the expression of Bcl-2,Bax,Caspase-3,ZO-1,Occludin,Claudin-1,and histone deacetylase 4.The levels of histone deacetylase in cell supernatant were detected by ELISA.The expression of histone deacetylase 4 in cells was detected by immunofluorescence.RESULTS AND CONCLUSION:(1)The cell viability of high sugar group was significantly lower than that of control group(P＜0.000 1).(2)The cells of the control group were in a good growth state,interwoven into a dense mesh,with interconnections between synapses.The cell growth of high glucose group was suppressed,and the connection of inter-cellular synapses was reduced.(3)Compared with the control group,the transmembrane resistance value of the high glucose group was reduced(P＜0.05);phenol-red permeability of the monolayer cell membrane was increased(P＜0.05);cell apoptosis rate was increased(P＜0.01);the expression of Bax protein was increased(P＜0.000 1);the expression of Caspase-3 protein had no significant change(P＞0.05);the expression of Bcl-2,ZO-1,Occludin,Claudin-1,and histone deacetylase 4 proteins was decreased(P＜0.01,P＜0.001,P＜0.01,P＜0.000 1,P＜0.01);the fluorescence expression of histone deacetylase 4 was decreased(P＜0.001)in the high glucose group.(4)The level of histone deacetylase 4 in the cell supernatant of the high glucose group was lower than that of the control group(P＜0.05).The results show that high glucose induces the increased apoptosis and enhances permeability of hCEMCE/D3 cells,and its mechanism may be related to the decreased expression level of histone deacetylase 4.

Objective To understand the trends in availability and usage for four non-statin lipid-lowering drugs between 2020 and 2023 in China,and to explore the impact of centralized drug procurement and national drug price negotiation policy on the accessibility and utilization trends of these medications.Methods Data processing was performed on four non-statin lipid-lowering drugs(ezetimibe,hybutimibeh,evolocumab,alirocumab)from over 1 000 hospitals within the China Pharmaceutical Association network units from 2020 to 2023 was performed on.SPSS 27.0 was employed for descriptive statistical analysis and x2 tests regarding drug availability,sales revenue,defined daily dose system(DDDs),and cost per DDD(DDDc).Results The availability and DDDs of four cholesterol absorption inhibitors or PCSK9-mAbs increased year by year during the study period,with the most significant increase observed in the first year after centralized drug procurement or national drug price negotiation policy.After the national drug price negotiation policy,alirocumab had the highest availability among the three negotiated drugs.The sales volume of ezetimibe,which was included in the centralized drug procurement program,increased before the negotiation but decreased significantly afterwards,with its share of sales declining year by year.During the study period,the sales volumes and market shares of the three negotiated drugs all increased annually,with evolocumab showing notably higher sales values and percentages than both hybutimibe and alirocumab after the negotiation.The DDDc for all four drugs continued to decrease during the survey period,with the first-year post-centralized drug procurement or national drug price negotiation policy witnessing the largest drop.Conclusion Both of the centralized drug procurement and the national drug price negotiation policy can significantly increase the accessibility to non-statin lipid-lowering drugs and promote their clinical application,however,there is a mismatch between the growth in usage and accessibility for certain varieties.

OBJECTIVE To investigate the effects of the split formulas of Biyuan Heji(BYHJ)on paranasal sinus mucosal in-flammation in rats with acute rhinosinusitis(ARS)based on the pathogenesis of"wind-cold transforming into lung-heat".METHODS Unilateral nasal cavity occlusion combined with nasal dripping of Staphylococcus aureus were performed to establish a rat model of ARS.SD rats were randomly divided into blank,model,BYHJ(wind-cold removal+lung-heat removal),lung-heat removal,wind-cold removal,and positive drug groups,with 6 rats in each group.The rats were treated with the corresponding drugs for 7 d and then the samples were collected.HE staining was used to observe the pathological changes of rat paranasal sinus mucosa tissues,ELISA was employed to determine the levels of interleukin-1β(IL-1β),IL-6,IL-8,IL-9,IL-10,and IL-12 in serum,immunohistochemis-try(IHC)was adopted to measure the protein expression of tumor necrosis factor-α(TNF-α)and intercellular adhesion molecule(ICAM-1)in paranasal sinus mucosa tissues,and Western blot was used to detect the protein expression of phosphorylated p38 mito-gen-activated protein kinase(p38 MAPK),nuclear transcription factor-κB p50(NF-κB p50),and NF-κB p65 in paranasal sinus mucosa tissues.RESULTS The acute sinusitis rat inflammation model was successfully established.Compared with the model group,the water drinking,diet eating,and body weight of rats in the BYHJ group,wind-cold removal,lung-heat removal,and positive drug groups were significantly improved,the aggregation of inflammatory cells in the paranasal sinus mucosal tissue was reduced,and the levels of IL-1β,IL-6,IL-8,IL-9,and IL-12 in the serum were significantly reduced(P<0.01),IL-10 content significantly in-creased(P<0.01),the protein expression of TNF-α,ICAM-1,p38 MAPK,NF-κB p50,and NF-κB p65 in the paranasal sinus mucosa was significantly decreased(P<0.01).The comparison between various traditional Chinese medicine groups showed that the decrease of IL-1β,IL-6,IL-8,IL-9,IL-12,TNF-α,ICAM-1,p38 MAPK,NF-κB p50,and NF-κB p65 and the increase of IL-10 in the BYHJ group were better than those in the split formula groups(P<0.01),and the lung-heat removal group was better than the wind-cold removal group(P<0.01).CONCLUSION BYHJ and its split formulas can effectively inhibit the inflammatory response in rats with ARS.

Objective To explore the efficacy of precise surface positioning method in percutaneous screw fixation for Jones fracture.Methods Data of 46 patients with Jones fractures admitted to our hospital from September 2021 to December 2023 were retrospectively analyzed.According to the different treatment methods,the patients were divided into screw groups(24 patients fixed by percutaneous screws)and tension band groups(22 patients fixed by incision and reset tension band).The general conditions,follow-up time,surgical time,hospitalization time,fracture reduction quality,postoperative complete weight-bearing time,fracture healing time,and complications in the two groups were recorded and compared.The American Association of Foot and Ankle Surgery(AOFAS)system was used to assess the recovery of foot function.Results There was no statistically significant difference in the general data before the operation between the two groups of patients(P＞0.05).The follow-up time of the two groups was 6 to 24 months.Compared with the tension band group,the screw group had shorter operation time[(32.96±5.35)min vs.(47.27±9.42)min],shorter hospitalization time[(5.75±1.18)d vs.(7.68±2.05)d],earlier postoperative complete weight-bearing time[(6.47±0.73)w vs.(7.29±1.66)w],faster fracture healing time[(3.31±0.68)months vs.(4.29±1.29)months],higher AOFAS score[(89.13±2.62)vs.(79.59±4.49)](all P＜0.05).The tension band group of fracture reduction quality was better than the screw group(P＜0.05).The quality of fracture reduction during the operation in the tension band group was better than that in the screw group(P＜0.05).In terms of complications:no complications occurred in the screw group,one patient in the tension band group developed infection,and one patient had a loose Kirchner needle.Conclusion Precise surface positioning method in percutaneous screw fixation for Jones fracture is a safe and effective method with precise positioning,simple operation,small trauma,and rapid rehabilitation,which is worthy of clinical promotion,but the indications need to be strictly controlled according to the type of fracture.

Objective To detect diatoms in tissues and water samples from drowning cases by gene array,and explore the application value of DNA microarray.Methods Diatoms in the lung,liver,kidney samples,and on-site water samples from 19 drowned corpses that appeared in the rivers of Zhongshan City between July 2021 and April 2022,were detected by gene arrays.Moreover,diatom detection was also performed on 28 samples from 7 corpses by microscope.Then the testing results were compared and analyzed.Results For those 28 samples,diatom types in both tissue and water samples by gene detection were more than those detected by microscopy,the detection was statistically significant(P<0.05).However,there was no statistically significant difference(P>0.05)in the detection of diatom types between tissue samples and water samples both using gene arrays.In terms of tissue samples,diatoms were easier to be detected in lung than liver or kidney,and there were also more types of diatoms detected in lung.Diatom detection by gene arrays showed that the diatom types showed a decreasing trend as the tissue was further away from the entrance of body,and the difference of diatom types increased among tissues.Conclusion The gene array can effectively detect the diatom types and distribution characteristics in drowned corpses,and has great potential in the research of the traceability analysis of corpses found in water and the causes of death.

The ribosomal protein S11(RPS11)from Enterococcus faecalis is was heterologously ex-pressed using the Pichia pastoris system.The RPS11 gene sequence was optimized to match the yeast codon preference,and the recombinant expression vector pPIC9K-RPS11 was constructed.Electroporation was used to transform the vector into the Pichia pastoris GS115 strain,and high-copy recombinant strains were selected through G418 resistance screening.Protein expression was induced with methanol,and the expression was verified by SDS-PAGE.The recombinant protein was applied in a mouse melanoma treatment model to evaluate its therapeutic effects.The results showed that the recombinant expression vector pPIC9K-RPS11 successfully expressed the target protein with an approximate molecular weight of 14 kDa in Pichia pastoris.The optimal fermenta-tion conditions were determined to be an induction temperature of 30 ℃,induction time of 72 h,and methanol concentration of 1％.Analysis using a mouse peritoneal macrophage trained immuni-ty model revealed that recombinant RPS11 possessed biological activity capable of inducing trained immunity.Additionally,therapeutic experiments in a mouse melanoma model demonstrated that recombinant RPS11 significantly inhibited tumor growth.These findings suggest that the recombi-nant RPS11 secreted by Pichia pastoris not only possesses biological activity in inducing trained immunity but also inhibits tumor cell growth in a mouse melanoma model,providing theoretical support for the heterologous expression and potential applications of recombinant RPS11.