Objective To explore the effects of Ginkgo biloba extract (GBE) on cardiac microvascular endothelial cells (CMECs) under oxygen and glucose deprivation/reperfusion (OGD/R) condition and its molecular mechanisms. Methods An OGD/R-induced injury model was established in CMECs. According to different intervention, CMECs were divided into four groups: normoxia blank control group (WT group), WT + GBE group, OGD/R group, and OGD/R + GBE group. Cell apoptosis was detected by flow cytometry technology in each group. The oxidative stress was examined by MitoSox staining. The migration abilities were measured by scratch assay. The expressions of PERK/eIF2α/CHOP, nuclear factor kappa B (NF-κB), and endothelial cell function markers were detected by Western blotting. Results Compared with the WT group, the endothelial cell apoptosis level in the OGD/R group significantly increased, with markedly aggravated cellular dysfunction. The expressions of p-NF-κB, vascular cell adhesion molecule-1 (VCAM-1), and intercellular cell adhesion molecule-1 (ICAM-1) were significantly upregulated (P＜0.05), and the activation of the CHOP signaling pathway was notably enhanced (P＜0.05). After intervention with GBE, endothelial cell apoptosis caused by OGD/R injury was significantly reduced, oxidative stress and inflammation levels were markedly downregulated, and the expression of p-NF-κB was considerably decreased (P＜0.05), while the CHOP signaling pathway was notably inhibited (P＜0.05). Furthermore, it was found that GBE could promote expression of SIRT6 to regulate the above molecules, thereby alleviating cardiac microvascular endothelial cell injury under OGD/R condition. On the contrary, when SIRT6 was knocked down, the protective effects were significantly reduced. Conclusions GBE improves endothelial cell dysfunction, endoplasmic reticulum stress, and endothelial cell apoptosis caused by OGD/R injury by promoting the expression of SIRT6 protein, thus regulating the NF-κB inflammatory pathway and CHOP signaling pathway.

Background::Cerebrovascular disease (CVD) ranks among the foremost factors responsible for mortality on a global scale. The mortality patterns of CVDs and temporal trends in China need to be well-illustrated and updated.Methods::We collected mortality data on patients with CVD from Chinese Center for Disease Control and Prevention’s Disease Surveillance Points (CDC-DSP) system. The mortality of CVD in 2020 was described by age, sex, residence, and region. The temporal trend from 2013 to 2019 was evaluated using joinpoint regression, and estimated rates of decline were extrapolated until 2030 using time series models.Results::In 2019, the age-standardized mortality in China (ASMRC) per 100,000 individuals was 113.2. The ASMRC for males (137.7/10 5) and rural areas (123.0/10 5) were both higher when stratified by gender and urban/rural residence. The central region had the highest mortality (126.5/10 5), the western region had a slightly lower mortality (123.5/10 5), and the eastern region had the lowest mortality (97.3/10 5). The age-specific mortality showed an accelerated upward trend from aged 55-59 years, with maximum mortality observed in individuals over 85 years of age. The age-standardized mortality of CVD decreased by 2.43% (95% confidence interval, 1.02-3.81%) annually from 2013 to 2019. Notably, the age-specific mortality of CVD increased from 2013 to 2019 for the age group of over 85 years. In 2020, both the absolute number of CVD cases and the crude mortality of CVD have increased compared to their values in 2019. The estimated total deaths due to CVD were estimated to reach 2.3 million in 2025 and 2.4 million in 2030. Conclusion::The heightened focus on the burden of CVD among males, rural areas, the central and western of China, and individuals aged 75 years and above has emerged as a pivotal determinant in further decreasing mortalities, consequently presenting novel challenges to strategies for disease prevention and control.

Aim To investigate the effect of quercetin(Que)on podocyte inflammatory injury and the under-lying mechanism.Methods MPC5 cells were divided into normal glucose group(NG),mannitol group(MA),high glucose group(HG)and high glucose+quercetin group(HG+Que).Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry.The expression of SIRT1,STAT3,apoptosis-related proteins(Bax,Bcl-2,caspase-3)and pyroptosis pro-tein GSDME was detected by Western blot.The ex-pression levels of inflammatory factors(IL-6,TNF-α,IL-18,IL-1β)in cell supernatants were detected by ELISA.Then small interfering RNA technology was used to knockdown SIRT1 expression.To further eval-uate the biological significance of SIRT1 in response to high glucose and Que treatment,negative control group(HG+si-NC+Que)and SIRT1 interference group(HG+si-SIRT1+Que)were added in the presence of high glucose and Que.Results Compared with the high glucose group,40 μmol·L-1 Que could alleviate the apoptosis of MPC5 cells induced by high glucose,decrease the expression of apoptosis related protein Bax and caspase-3,as well as increase the expression of anti-apoptotic protein Bcl-2;ELISA results showed that Que could decrease the expression of TNF-α,IL-6,IL-1 β and IL-18 induced by high glucose.Mechanical-ly,Que could alleviate the inhibitory effect of high glu-cose on the expression of SIRT1,and further decrease the activation of STAT3 and N-GSDME,and inhibit pyroptosis.Compared with the si-NC group,si-SIRT1 group could reverse the protective effect of Que on the high glucose induced inflammatory damage of podo-cytes,the expression of apoptotic proteins Bax and caspase-3 increased,while the expression of anti-apop-totic protein Bcl-2 decreased.At the same time,the levels of inflammatory cytokines TNF-α,IL-6,IL-1 βand IL-18 in supernatants increased,and the expres-sion of STAT3 and N-GSDME increased.Conclusion Que could inhibit pyroptosis and relieve the inflam-matory damage of podocytes through SIRT1/STAT3/GSDME pathway.

Objective:To standardize the bacterial endotoxin testing criteria for zoledronic acid injection and estab-lish a detection method using recombinant factor C(rFC).Methods:The gel-clot method(BET)was utilized to test 13 batches of zoledronic acid injection from national supervision and random inspection.Interference tests were conducted on zoledronic acid injections from three manufacturers at different concentrations(500,100,50,25 μg·mL-1)using rFC test kits from two manufacturers.Results:Detection was performed for the specification of 100 mL∶5 mg and other specifications according to＜0.50 EU per 1 mL and＜5.0 EU per 1 mg,respectively,and all results met the criteria.The recovery rate for 25 μg·mL-1 using rFC kits from both manufacturers ranged between 50％and 200％.Validation of rFC.Methods:Eight batches of zoledronic acid injection were validated at 25 μg·mL-1,five batches of zoledronic acid injection(100 mL∶5 mg)were validated at 0.5 EU·mL-1,and all recovery rates were between 50％and 200％.Conclusion:The bacterial endotoxin testing method for zoledronic acid injection can be established as follows:for large volume injection products with 100 mL or more,each 1 mL should contain less than 0.50 EU of endotoxin(following the Chinese Pharmacopoeia 2020,general chapter 1143).For other specifications,each 1 mg of zoledronic acid should contain less than 10.0 EU of endotoxin.The rFC test kit method for bacterial endotoxins involved diluting the sample with water for bacterial endotoxin testing to contain 25 μg of zoledronic acid per 1 mL,or performing 1∶1 dilution for large volume injections(100 mL∶5.0 mg specifi-cation),with recovery rate between 50％and 200％as per the kit instructions.

Objective:To detect the expression of L-type amino acid transporter 1(LAT1)in non-Hodgkin's lymphoma(NHL)tissues,and analyze its effect on clinicopathological characteristics and prognosis of patients.Methods:A total of 92 NHL patients who were treated in our hospital from January 2017 to April 2019 were collected.The expression of LAT1 in NHL tissue was detected by immunohistochemistry and compared between patients with different pathological features(including sex,Ann Arbor stage,extranodal infiltration,Ki-67).The risk factors affecting mortality were analyzed using univariate and multivariate Cox proportional hazards regression.Receiver operating characteristic(ROC)curve was used to detect the predictive value of percentage of LAT1-positive cells in NHL tissue for patient mortality,and analyzing the effect of percentage of LAT1-positive cells on survival rate.Results:LAT1 was positively expressed in NHL tissue.The high expression rate of LAT1 in Ann Arbor stage Ⅲ and Ⅳ groups were higher than that in Ann Arbor stage Ⅰ group,that in extranodal infiltration group was higher than non-extranodal infiltration group,and that in Ki-67 positive expression group was higher than Ki-67 negative expression group(all P＜0.05).The remission rate after 3 courses of treatment in high-LAT1 expression group was 70.7％,which was lower than 91.2％in low-LAT1 expression group(P＜0.05).Ann Arbor stage Ⅲ and Ⅳ,extranodal invasion,Ki-67 positive expression and increased expression of LAT1(LAT1-positive cell percentage score ≥ 2)were risk factors for mortality.The cut-off value of percentage of LAT1-positive cells for predicting NHL death was 45.6％,and the area under the ROC curve was 0.905(95％CI:0.897-0.924).The 3-year survival rate of high-LAT1 level group(the percentage of LAT1-positive cells ≥ 45.6％)was 50.00％,which was lower than 78.26％of low-LAT1 level group(P＜0.05).Conclusion:The expression level of LAT1 in NHL tissue increases,which affects Ann Arbor stage and extranodal infiltration of patients.LAT1 is a risk factor for death.

Objective:To investigate the effect of feeder layer cells expressing interleukin(IL)-21 on the amplification of NK cells in vitro.Methods:The K562 cell line with IL-21 expression on its membrane was constructed by electroporation,and co-cultured with NK cells after inactivation.The proliferation of NK cells was observed.The killing function of the amplified NK cells in vitro was evaluated by the lactate dehydrogenase(LDH)and interferon-γ(IFN-y)release assay.A colorectal cancer xenograft model in NOD/SCID mice was established,and a blank control group,a NK cell group and an amplified NK cell group were set up to detect the tumor killing effect of amplified NK cells in vivo.Results:K562 cells expressing IL-21 on the membrane were successfully constructed by electroporation.After co-culturing with K562 cells expressing IL-21 on the membrane for 17 days,the NK cells increased to 700 times,which showed an enhanced amplification ability compared with control group(P＜0.001).In the tumor cell killing experiment in vitro,there was no significant difference in the killing activity on tumor cells between NK cells and amplified NK cells,and there was also no significant difference in mice in vivo.Conclusion:K562 cells expressing IL-21 on the membrane can significantly increase the amplification ability of NK cells in vitro,but do not affect the killing function of NK cells in vitro and in vivo.It can be used for the subsequent large-scale production of NK cells in vitro.

Objective:To perform harmonization based on the ComBat method for PET brain imaging scanned by different types of scanners from the same manufacturer and explored its effect on center effect.Methods:The three-dimensional (3D) Hoffman brain model was scanned by two different PET/CT instruments (Siemens Biograph64 TruePoint and Biograph128 mCT). Fourteen healthy subjects (8 males, 6 females, age: (57.7±9.5) years) underwent 18F-FDG PET/CT on Siemens Biograph64 TruePoint and 12 healthy subjects (9 males, 3 females, age: (55.8±10.5) years) underwent 18F-FDG PET/CT on Siemens Biograph128 mCT (all from Huashan Hospital, Fudan University; from November 2020 to March 2023). The whole brain was divided into 116 brain regions based on the anatomical automatic labeling (AAL) brain template. The ComBat method was applied to harmonized the PET data from brain model and healthy subjects. Mann-Whitney U test was performed on the radioactive counts and SUV ratios (SUVR) before and after homogenization acquired by both PET/CT instruments. Voxel-based statistical parametric mapping (SPM) independent-sample t test was also performed on data of healthy subjects. Results:In 3D Hoffman brain model, radioactivity counts (5 590.33(4 961.67, 6 102.95) vs 6 116.03(5 420.97, 6 660.66); z=-9.35, P<0.001) and SUVR (1.35(1.19, 1.47) vs 1.37(1.21, 1.49); z=-3.63, P<0.001) were significantly different between the two PET/CT scanners before harmonization and not after harmonization (radioactivity counts: 5 845.95(5 192.68, 6 378.63) vs 5 859.17(5 193.84, 6 380.52); SUVR: 1.35(1.20, 1.48) vs 1.36(1.20, 1.49); both z=-0.68, both P=0.498). In the healthy subjects, radioactive counts in 19 brain regions (12 422.78(11 181.60, 13 424.28)-18 166.40(15 882.80, 18 666.27); z values: from -3.24 to -2.06, all P<0.05) and SUVR in 40 brain regions (1.46(1.41, 1.52)-2.28(2.16, 2.36); z values: from -3.65 to -1.70, all P<0.05) were significantly different between the two scanners before harmonization, while after homogenization there were no statistical differences for all 116 brain regions (radioactivity counts: 9 243.55(8 502.38, 9 854.87)-20 419.60(19 931.51, 21 179.43); z values: from -0.72 to 0, all P>0.05; SUVR: 1.04(1.01, 1.09)-2.32(2.24, 2.40); z values: from -0.82 to 0, all P>0.05). SPM showed that significant differences of glucose metabolism in the cerebral cortex, basal ganglia, midbrain and cerebellum were found in healthy subjects between the two PET/CT scanners before homogenization, and brain regions with obvious differences reduced after homogenization. Conclusion:ComBat harmonization method is efficient at removing the center effect among different types of PET/CT scanners from the same manufacturer and may provide a simple and easy-to-implement homogenization for multicenter brain imaging studies.

AIM To study the chemical constituents from the branches and leaves of Toona ciliata Roem.var.pubescens(Franch.)Hand-Mazz.and their antitumor activities.METHODS The compounds were isolated and purified by silica gel,RP-18 reverse phase silica gel and semi-preparative HPLC,the structures of compounds were identified by physicochemical properties and spectral data.The antitumor activities were determined by MTT method.RESULTS Fifteen compounds were isolated and identified as toonaolide D(1),toonaciliatin E(2),bourjotinolone A(3),(21R,23R)-epoxy-21α-ethoxy-24S,25-dihydroxyapotirucalla-7-en-3-one(4),(Z)-toonasterone C(5),(E)-toonasterone(6),3-epi-dyscusin C(7),(Z)-aglawone(8),(E)-volkendousin(9),8(14),15-isopimaradiene-2α,3α,19-triol(10),(-)-loliolide(11),cyclohexenone(12),pubinernoid A(13),quercetin-3-O-(4″-methoxy)-α-L-rahmnopyranosyl(14),5-hydroxymethyl-2-furancarboxaldehyde(15).The IC50 values of compounds 3 and 4 on K562 cells were 54.2 and 47.3 μmol/L,respectively,and the IC50 values on HEL cells were 47.3 and 61.1 μmol/L,respectively.CONCLUTION Compounds 4,7,10 and 11 are isolated from Toona genus for the first time,and compounds 2,15 are first isolated from this plant.Compounds 3 and 4 show weak antitumor activities.

Multimorbidity management for the elderly is a big challenge for China′s healthcare system and the people centered integrated care (PCIC) is a new concept in health care reform, and the application of the PCIC in multimorbidity management can better improve the health servece for elderly patients. This article applies SWOT analysis to assess the internal strengths and weaknesses as well as external opportunities and threats of applying the PCIC to the geriatric management of multimorbidity; also proposes strategies of geriatric management for elderly patients with Chinese characteristics based on concept of PCIC. The proposed stretegies optimize the personalized assessment with the digital technology, promotes the management with close-knit healthcare system, and strengthen the role of primary medical institutions to improve the long-term management mechanism of multimorbidity.

Objective:To screen key genes that co-regulate immune and mitochondrial energy metabolism through bioinformatics methods and to investigate the relationship between the key genes and immune infiltration.Methods:A total of 671 glioma samples (the tumor group) and 5 non-tumor brain tissue samples (the control group) were collected from the Cancer Genome Atlas (TCGA) database on November 13, 2023. Through a comprehensive search of the GeneCards database and immune-related genes (IRG) and mitochondrial energy metabolism-related genes (MEMRG) in previous published literatures, 76 IRG and MEMRG (IR & MEMRG) were obtained by taking the intersection of IRG and MEMRG after merging and deduplicating. The limma package in R software was used to screen the differentially expressed genes (DEG) between the tumor group and the control group. Then, immune-related & mitochondrial energy metabolism-related differentially expressed genes (IR&MEMRDEG) were obtained by intersecting with IR & MEMRG. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted on the IR&MEMRDEG through the clusterProfiler package in R software. The STRINGv12.0 online database (https://cn.string-db.org/) was employed to construct a protein interaction network based on IR&MEMRDEG and to identify the top 5 key core genes. Single-sample gene-set enrichment analysis (ssGSEA) was used to determine the relative abundance of immune cell infiltration in all samples, and the immune cell infiltration matrices for both the tumor and the control groups were acquired. The expression differences in infiltration abundance of the immune cells in the tumor group and the control group were analyzing by using the ggplot2 package in R software. The heat map was drawn by utilizing the R software pheatmap package to show self-correlation of immune cells. The correlation between the top 5 key genes in the protein interaction network and immune cells was calculated by using the Spearman algorithm and the R software ggplot2 package.Results:A total of 3 623 DEGs were identified from the TCGA database in both groups, including 1 711 up-regulated genes and 1 912 down-regulated genes. After taking the intersection of DEG and IR&MEMRG, 11 IR&MEMRDEG were obtained including EIF4EBP1, TP53, IDH1, PRCKZ, CD200, GPI, PGM2, PKLR, AK2, ATP4A, and ALDH3B1. GO enrichment analysis results showed that 11 IR&MEMRDEG were mainly enriched in ADP metabolic process, ATP metabolic process, purine nucleoside diphosphate metabolic process, purine ribonucleoside diphosphate metabolic process, and ribonucleoside diphosphate metabolic process at the biological level; in the fibronectin-1 rich granule, secretory granule lumen, cytoplasmic vesiclelumen, vesiclelumen, and nuclear matrix at the cellular component level; in magnesium ion binding, potassium ion binding, and alkali metal ion binding at the molecular functional level. The KEGG enrichment analysis results showed that 11 IR&MEMRDEGs were mainly enriched in glycolysis/gluconeogenesis, carbon metabolism, insulin signaling pathway, pentose phosphate pathway, starch and sucrose metabolism signaling pathways. The protein interaction network analysis from the STRING database revealed that 5 highest scoring core proteins were identified, namely EIF4EBP1, TP53, IDH1, PRKCZ, and AK2.The immune infiltration abundances of 28 immune cells were calculated by using the ssGSEA algorithm. The infiltration abundance of 15 immune cells in the tumor group was higher than that in the control group, and the differences were statistically significant (all P < 0.05). The findings from the immune infiltration analysis indicated a positive correlation among 15 types of immune cells, in which there was a strongest correlation between effector memory CD8 + T cell and myeloid derived suppressor cells. EIF4EBP1, TP53, IDH1, and AK2 exhibited a positive correlation with a large number of immune cells (all P < 0.05), whereas PRKCZ demonstrated a negative correlation with more immune cells (all P < 0.05). Conclusions:PRKCZ, AK2, and EIF4EBP1 have the potential to be the new targets of immunotherapy for gliomas.