The Proctor's reporting guideline for implementation strategies represents a landmark framework in the field of implementation science, aiming to address the issue of inconsistent reporting in implementation research by standardizing the naming, definition, and operationalization of implementation strategies, thereby enhancing the credibility and utility of research findings. This paper provides an in-depth interpretation of the core connotations of this reporting guideline and illustrates its application in developing interview outlines and specifying implementation strategies, using a brief smoking cessation intervention project as a case study. Through this reporting guideline, abstract recommendations for implementation are systematically transformed into clear, multidimensional operational guides, significantly improving the transparency of strategy connotations and the replicability of actual execution. Meanwhile, the case study highlights the flexibility of the guideline, which allows researchers to adapt the content and format of strategies based on local resources and cultural contexts, thus enhancing practical adaptability while maintaining scientific rigor. However, the application of Proctor's reporting guideline still faces challenges, primarily manifested in the potential confusion surrounding the constructs of temporality and dose in practice, as well as the challenges that the inherent flexibility of the guideline may pose to the assessment of fidelity and effectiveness. Despite these limitations, the reporting guideline remains a vital tool for implementation research; future efforts should focus on optimizing its application—through refining operational guidelines, standardizing flexible adaptations, and involving stakeholders—to better guide implementation studies and continuously promote high-quality development in the field.

Objective: To analyze the association between dietary diversity and cognitive function among the elderly using group-based trajectory model, so as to provide the basis for formulating dietary intervention strategies to prevent cognitive impairment. Methods: Based on the Chinese Longitudinal Healthy Longevity Survey (CLHLS) project, demographic information, lifestyle behaviors, psychological well-being, and activities of daily living of elderly individuals aged ≥65 years from 2008 to 2018 were collected. Dietary diversity was assessed using a food frequency questionnaire, with a score of <7 points defined as low dietary diversity. Cognitive function was evaluated using the Chinese version of the Mini-Mental State Examination (MMSE). A group-based trajectory model was established based on cognitive function scores from 2008 to 2018 to analyze the trajectories of cognitive function change. The association between dietary diversity and cognitive function was analyzed using a multinomial logistic regression model. Results: A total of 1 613 individuals were collected, with a median age was 72.00 (interquartile range, 10.00) years. There were 810 males (50.22%) and 803 females (49.78%). The group-based trajectory model analysis categorized the participants into three groups: the low-level normal group, the high-level normal group, and the slow-then-rapid decline group, comprising 796 (49.35%), 585 (36.27%), and 232 (14.38%) individuals, respectively. Among these groups, the numbers of individuals with low dietary diversity were 497 (62.44%), 311 (53.16%), and 166 (71.55%), respectively, with a statistically significant difference (P<0.05). Multinomial logistic regression analysis showed that after adjusting for demographic information, lifestyle behaviors, psychological well-being, and activities of daily living, compared with the high-level normal group, low dietary diversity was statistically associated with cognitive function in the slow-then-rapid decline group (OR=1.622, 95%CI: 1.103-2.384). Conclusion Low dietary diversity may increase the risk of cognitive impairment among the elderly.

Objective To investigate the levels of vascular endothelial growth factor(VEGF)and vas-cular endothelial growth factor receptor 2(VEGFR2)in serum and urine of the children patients with primary nephrotic syndrome(PNS),and to conduct the correlation analysis on the experimental indexes of 24 h urine protein quantitation,blood albumin(ALB),total cholesterol(TC),blood urea nitrogen(BUN)and creatinine(Scr).Methods Thirty children patients with PNS without hormone treatment admitted and treated in this hospital from January 2020 to December 2023 were selected as the study subjects and divided into the hormone sensitive nephrotic syndrome(SSNS)group(n=12)and steroid dependent nephrotic syndrome(SDNS)group(n=18)according to the cortical hormone treatment effect.Twenty healthy children with matched age and sex visiting in the pediatric department during the same period served as the control group.The differ-ences of VEGF and VEGFR2 levels in blood and urine were compared among 3 groups and their correlation with the experimental indexes was analyzed.Results Compared with the control group,the blood VEGF lev-el,urine VEGF and BUN levels in the SDNS group were increased,blood VEGFR2 and urine VEGFR2,24 h urine protein quantitation and TC level in the SSNS group and SDNS group were increased,the ALB level was decreased,and the differences were statistically significant(P<0.05).The Pearson correlation analysis re-sults showed that blood VEGF level was positively correlated with the 24 h urine protein quantitation and TC level,and negatively correlated with the ALB level(P<0.05);the urine VEGF level was positively correlated with 24 urine protein quantitation and TC level,and negatively correlated with the Scr level(P<0.05);blood VEGF2 level was positively correlated with the 24 h urine protein quantitation and the TC level,and negative-ly correlated with the ALB level(P<0.05);the urine VEGF2 level was positively correlated with the 24 h u-rine protein quantitation and TC level,and negatively correlated with the ALB level(P<0.05).Conclusion The blood and urine VEGF and VEGFR2 may serve as the novel indicators for the early diagnosis,prediction and hormone sensitivity evaluation in the children patients with PNS.

Objective:To establish auto verification rules for the routine coagulation assays,and to provide reference for clinical laboratories to improve the quality and efficiency of results verification.Methods:A total of 24,510 specimens of sodium citrate anticoagulation routine coagulation test from the laboratory departments of eight hospitals including the First Medical Center,Chinese PLA General Hospital during January to March 2020 were collected and randomly divided into a rule establishment group and a rule verification group,with 6,670 specimens in the rule establishment group,including 2,056 Delta checks,and 17,840 specimens in the rule validation group,including 3,210 Delta checks.The activities of prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(Fib),thrombin time(TT),D-dimer(DD)and/or antithrombin(AT)were detected by Stago STA R Max automatic coagulation analyzer and supporting reagents.Taking the manual verification results as the standard,the auto verification and manual false negative rate(invalid verification),false positive rate(invalid interception),pass rate,positive coincidence rate,negative coincidence rate,verification consistency rate and specimen turnaround time(TAT)of the two groups were calculated.Results:The auto verification rules and the application process were preliminarily established,including internal quality control,alarm information,auto verification scope,critical value and deviation value inspection.In the rule establishment group,the single item pass rate was 82.6%-92.4%,and the overall pass rate was 73.8%.The consistency rate between auto verification and manual verification was 98.2%,and the positive coincidence rate and negative coincidence rate were 24.4%and 73.8%,respectively.In the rule verification group,the single item pass rate was 86.4%-91.5%,and the overall review pass rate was 71.5%.By simulating the application of auto verification rules,the average TAT of two hospitals among the eight hospitals was shortened by 1.5 hours and 2.1 hours,respectively.Conclusion:The application of auto verification rules can reduce workload of manual verification,and significantly shorten the TAT,and improve the report efficiency of the laboratory.

Background: Evidence has revealed the involvement of microRNAs (miRNAs) in modulating osteogenic differentiation, implying the promise of miRNA-based therapies for treating osteoporosis. This study investigated whether miR-181a-5p influences osteogenic differentiation and bone formation and aimed to establish the mechanisms in depth. Methods: Clinical serum samples were obtained from osteoporosis patients, and MC3T3-E1 cells were treated with osteogenic induction medium (OIM) to induce osteogenic differentiation. miR-181a-5p-, Runt-related transcription factor 1 (Runx1)-, and/or allograft inflammatory factor-1 (AIF-1)-associated oligonucleotides or vectors were transfected into MC3T3-E1 cells to explore their function in relation to the number of calcified nodules, alkaline phosphatase (ALP) staining and activity, expression levels of osteogenesis-related proteins, and apoptosis. Luciferase activity, RNA immunoprecipitation, and chromatin immunoprecipitation assays were employed to validate the binding relationship between miR-181a-5p and Runx1, and the transcriptional regulatory relationship between Runx1 and AIF-1. Ovariectomy (OVX)-induced mice were injected with a miR-181a-5p antagonist for in vivo verification. Results: miR-181a-5p was highly expressed in the serum of osteoporosis patients. OIM treatment decreased miR-181a-5p and AIF-1 expression, but promoted Runx1 expression in MC3T-E1 cells. Meanwhile, upregulated miR-181a-5p suppressed OIM-induced increases in calcified nodules, ALP content, and osteogenesis-related protein expression. Mechanically, miR-181a-5p targeted Runx1, which acted as a transcription factor to negatively modulate AIF-1 expression. Downregulated Runx1 suppressed the miR-181a-5p inhibitor-mediated promotion of osteogenic differentiation, and downregulated AIF-1 reversed the miR-181a-5p mimic-induced inhibition of osteogenic differentiation. Tail vein injection of a miR-181a-5p antagonist induced bone formation in OVX-induced osteoporotic mice. Conclusion In conclusion, miR-181a-5p affects osteogenic differentiation and bone formation partially via the modulation of the Runx1/AIF-1 axis.

Post-stroke depression (PSD) is a common emotional disorder after stroke, which can affect cognitive function of patients and have adverse effect on post-stroke recovery. Diffusion tensor imaging (DTI) can reveal the microstructure of white matter. At present, it has been applied in the study of the pathogenesis of various diseases. This article reviews the application of DTI technology in depression, stroke, and PSD, aiming to make early predictions of PSD from multiple perspectives to improve its outcomes.

OBJECTIVES: The intestinal microbial characteristics of patients with simple cerebral infarction (CI) and CI complicated with Type 2 diabetes mellitus (CI-T2DM) are still not clear. This study aims to analyze the differences in the variable characteristics of intestinal flora between patients simply with CI and CI-T2DM. METHODS: This study retrospectively collected the patients who were admitted to the Affiliated Hospital of Putian University from September 2021 to September 2022. The patients were divided into a CI group (n=12) and a CI-T2DM group (n=12). Simultaneously, 12 healthy people were selected as a control group. Total DNA was extracted from feces specimens. Illumina Novaseq sequencing platform was used for metagenomic sequencing. The Knead Data software, Kraken2 software, and Bracken software were applied for sequencing analysis. RESULTS: At phylum level, the average ratio of Firmicutes, Bacteroidetes, and Proteobacteria in the CI-T2DM group were 33.07%, 54.80%, and 7.00%, respectively. In the CI group, the ratios of each were 14.03%, 69.62%, and 11.13%, respectively, while in the control group, the ratios were 50.99%, 37.67%, and 5.24%, respectively. There was significant differences in the distribution of Firmicutes (F=6.130, P=0.011) among the 3 groups. At the family level, compared with the CI group, the relative abundance of Eubacteriaceae (t=8.062, P<0.001) in the CI-T2DM group was significantly increased, while Corynebacteriaceae (t=4.471, P<0.001), Methanobacteriaceae (t=3.406, P=0.003), and Pseudomonadaceae (t=2.352, P=0.028) were decreased significantly. At the genus level, compared with the CI group, there was a relative abundance of Cutibacterium (t=6.242, P<0.001), Eubacterium (t=8.448, P<0.001), and Blautia (t=3.442, P=0.002) in the CI-T2DM group which was significantly increased. In terms of Methanobrevibacter (t=3.466, P=0.002), Pyramidobacter (t=2.846, P=0.009) and Pseudomonas (t=2.352, P=0.028), their distributions were decreased significantly in the CI-T2DM group. At the species level, compared with the CI group, the relative abundance of Cutibacterium acnes (t=6.242, P<0.001) in the CI-T2DM group was significantly increased, while Pseudomonas aeruginosa (t=2.352, P=0.028) was decreased significantly. Still at the genus level, linear discriminant analysis effect size (LEfSe) analysis showed that the distributions of Pseudomonas and Blautia were determined to be the most significantly different between the CI-T2DM and the CI group. At the species level, the total number of operational taxonomic units (OTUs) in the 3 groups was 1 491. There were 169, 221, and 192 kinds of OTUs unique to the CI-T2DM, CI, and control group, respectively. CONCLUSIONS From phylum level to species level, the composition of intestinal flora in the patients with CI-T2DM is different from those in the patients simply with CI. The change in the proportion of Firmicutes, Bacteroidetes and Proteus compared with the healthy population is an important feature of intestinal flora imbalance in the patients with CI and with CI-T2DM. Attention should be paid to the differential distribution of Bacteroides monocytogenes and butyrate producing bacteria.
Humans ; Gastrointestinal Microbiome/genetics* ; Diabetes Mellitus, Type 2/complications* ; Retrospective Studies ; Bacteria/genetics* ; High-Throughput Nucleotide Sequencing

OBJECTIVES: To explore the mechanism of transforming growth factor-β1 (TGF-β1) induce renal fibrosis. METHODS: Renal fibroblast NRK-49F cells treated with and without TGF-β1 were subjected to RNA-seq analysis. DESeq2 was used for analysis. Differentially expressed genes were screened with the criteria of false discovery rate<0.05 and l o g 2 F C >1. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed for differentially expressed genes. Genes encoding transcription factors were further screened for differential expression genes. Then, the expression of these genes during renal fibrosis was verified using unilateral ureteral obstruction (UUO)-induced mouse renal fibrosis model and a public gene expression dataset (GSE104954). RESULTS: After TGF-β1 treatment for 6, 12 and 24 h, 552, 1209 and 1028 differentially expressed genes were identified, respectively. GO analysis indicated that these genes were significantly enriched in development, cell death, and cell migration. KEGG pathway analysis showed that in the early stage of TGF-β1 induction (TGF-β1 treatment for 6 h), the changes in Hippo, TGF-β and Wnt signaling pathways were observed, while in the late stage of TGF-β1 induction (TGF-β1 treatment for 24 h), the changes of extracellular matrix-receptor interaction, focal adhesion and adherens junction were mainly enriched. Among the 291 up-regulated differentially expressed genes treated with TGF-β1 for 6 h, 13 genes (Snai1, Irf8, Bhlhe40, Junb, Arid5a, Vdr, Lef1, Ahr, Foxo1, Myc, Tcf7, Foxc2, Glis1) encoded transcription factors. Validation in a cell model showed that TGF-β1 induced expression of 9 transcription factors (encoded by Snai1, Irf8, Bhlhe40, Junb, Arid5a, Vdr, Lef1, Myc, Tcf7), while the expression levels of the other 4 genes did not significantly change after TGF-β1 treatment. Validation results in UUO-induced mouse renal fibrosis model showed that Snai1, Irf8, Bhlhe40, Junb, Arid5a, Myc and Tcf7 were up-regulated after UUO, Vdr was down-regulated and there was no significant change in Lef1. Validation based on the GSE104954 dataset showed that IRF8 was significantly overexpressed in the renal tubulointerstitium of patients with diabetic nephropathy or IgA nephropathy, MYC was highly expressed in diabetic nephropathy, and the expressions of the other 7 genes were not significantly different compared with the control group. CONCLUSIONS TGF-β1 induces differentially expressed genes in renal fibroblasts, among which Irf8 and Myc were identified as potential targets of chronic kidney disease and renal fibrosis.
Mice ; Animals ; Humans ; Transforming Growth Factor beta1/metabolism* ; Diabetic Nephropathies/pathology* ; Transcriptome ; Signal Transduction ; Kidney ; Ureteral Obstruction/pathology* ; Fibrosis ; Interferon Regulatory Factors ; Transforming Growth Factor beta/metabolism* ; DNA-Binding Proteins/metabolism* ; Transcription Factors/metabolism*

Objective:To investigate the effect and mechanism of Acyl-CoA: lysocardiolipin acyltransferase 1 (ALCAT1) on hepatocyte steatosis and oxidative stress in fatty liver cell model.Methods:A fatty liver cell model was established and induced by free fatty acids (FFA). The expression of ALCAT1 in fatty liver cell model was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The empty siRNA plasmid and ALCAT1 siRNA plasmid were constructed. For the fatty liver cell model group, human normal hepatocytes (L-02 cells) were transfected with empty siRNA plasmid for 24 hours, and then cultured with FFA for 24 hours. For the ALCAT1 interfering group, L-02 cells were transfected with ALCAT1 siRNA plasmid for 24 hours, and then cultured with FFA for 24 hours. And L-02 cells cultured in common medium were used as as blank control group. Lipid droplet deposition and mitochondrial morphology were observed under transmission electron microscopy. The expression levels of autophagy-associated proteins (microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ and Beclin1) and key proteins of autophagy signal pathway (mammalian target of rapamycin (mTOR) and serine/threonine kinase (AKT)) were measured by Western blotting. The expression levels of oxidative stress products (malondialdehyde, 4-hydroxynonenal (4-HNE) and reactive oxygen species (ROS)) and inflammatory factors (interleukin-6(IL-6) and tumor necrosis factor (TNF)-α) were detected by enzyme-linked immunosorbent assay (ELISA) kits. Independent sample t test was used for statistical analysis. Results:The mRNA and protein expression levels of ALCAT1 of the fatty liver cell model group were both higher than that of negative control group (9.26±0.83 vs. 1.02±0.12, 0.35±0.02 vs. 0.17±0.01), and the differences were statistically significant ( t=9.82 and 6.83, both P<0.05). The results of electron microscopy indicated that the deposition of lipid droplets of the fatty liver cell model group and ALCAT1 interfering group were both higher than that of blank control group (17.67±3.52 and 7.67±0.33 vs. 4.33±0.33), the quantity of lipid droplets deposition of ALCAT1 interfering group was lower than that of fatty liver cell model group (7.67±0.33 vs. 17.67±3.52), and the differences were statistically significant ( t=3.76, 7.07 and 2.82, all P<0.05). The degree of mitochondria swelling of fatty liver cell model group was higher than that of blank control group and the degree of mitochondria swelling of ALCAT1 interfering group was lower than that of fatty liver cell model group. The results of Western blotting showed that the expression level of LC3-Ⅱof the fatty liver cell model group was higher than that of the blank control group (0.43±0.01 vs. 0.28±0.02), and the difference was statistically significant ( t=7.32, P<0.05). However there was no significant difference in the expression level of Beclin1 between fatty live cell model group and blank control group (0.93±0.05 vs. 0.98±0.05, P>0.05). The expression levels of LC3-Ⅱ and Beclin1 of the ALCAT1 interfering group were both higher than those of the fatty liver cell model group and blank control group (0.95±0.04 vs. 0.42±0.01 and 0.28±0.02, 2.07±0.06 vs. 0.93±0.05 and 0.98±0.05), and the differences were statistically significant ( t=13.30, 15.63, 14.05 and 13.02, all P<0.05). The expression levels of mTOR of the fatty liver cell model group and ALCAT1 interfering group were both lower than that of the blank control group (1.44±0.02 and 0.74±0.01 vs. 1.93±0.10), the expression level of mTOR of the ALCAT1 interfering group was lower than that of the fatty liver cell model group (0.74±0.01 vs. 1.44±0.02), and the differences were statistically significant ( t=4.83, 12.04 and 32.14, all P<0.05). The expression levels of phosphorylated AKT of the fatty liver cell model group and ALCAT1 interfering group were both lower than that of the blank control group (0.14±0.01 and 0.07±0.01 vs. 0.28±0.01), while the expression level of phosphorylated AKT of the ALCAT1 interfering group was lower than that of the fatty liver cell model group (0.07±0.01 vs. 0.14±0.01), and the differences were statistically significant ( t=8.59, 14.10 and 5.96, all P<0.05). The results of ELISA indicated that the expression levels of ROS, malondialdehyde, 4-HNE, IL-6 and TNF-α of the fatty liver cell model group and the ALCAT1 interfering group were all higher than those of the blank control group ((11.44±0.30) and (5.84±0.36) g/L vs. (1.72±0.38) g/L; (19.94±2.47) and (11.95±1.55) μmol/L vs. (1.47±0.18) μmol/L; (5.00±0.43) and (2.99±0.37) ng/L vs. (1.46±0.23) ng/L; (203.40±5.16) and (92.07±11.98) ng/L vs. (23.32±3.33) ng/L; (123.70±8.38) and (67.42±4.88) ng/L vs. (47.18±4.57) ng/L), and the differences were all statistically significant ( t=19.86, 7.86, 7.45, 6.74, 7.22, 3.49, 29.34, 5.53, 8.02 and 3.03, all P<0.05). While the expression levels of ROS, 4-HNE, IL-6 and TNF-α of the ALCAT1 interfering group were all lower than those of the fatty liver cell model group ((5.84±0.36) g/L vs. (11.44±0.30) g/L, (2.99±0.37) ng/L vs. (5.00±0.43) ng/L, (92.07±11.98) ng/L vs. (203.40±5.16) ng/L and (67.42±4.88) ng/L vs. (123.70±8.38) ng/L), and all the differences were statistically significant ( t=11.99, 3.51, 8.54 and 5.81, all P<0.05). There was no statistically significant difference in the expression of malondialdehyde between ALCAT1 interfering group and fatty liver cell model group ((11.95±1.55) μmol/L vs. (19.94±2.47) μmol/L, P>0.05). Conclusions:The expression of ALCAT1 is up-regulated in fatty liver cell model. Knockdown of ALCAT1 can inhibit the expression of mTOR pathway proteins, activate autophagy, alleviate hepatocyte steatosis, oxidative stress and inflammatory response.