CRISPR/Cas9-based therapeutics face significant challenges in penetrating the dense microenvironment of solid tumors, resulting in insufficient gene editing and compromised treatment efficacy. Current nanostrategies, which mainly focus on the paracellular pathway attempted to improve gene editing performance, whereas their efficiency remains uneven in the heterogenous extracellular matrix. Here, the nanoCRISPR system is prepared with self-cascading mechanisms for gene editing-mediated robust apoptosis and transcellular penetration. NanoCRISPR unlocks its self-cascade capability within the matrix metallopeptidase 2-enriched tumor microenvironment, initiating the transcellular penetration. By facilitating cellular uptake, nanoCRISPR triggers robust apoptosis in edited malignancies, promoting further transcellular penetration and amplifying gene editing in neighboring tumor cells. Benefiting from self-cascade between robust apoptosis and transcellular penetration, nanoCRISPR demonstrates continuous gene transfection/tumor killing performance (transfection/apoptosis efficiency: 1st round: 85%/84.2%; 2nd round: 48%/27%) and homogeneous penetration. In xenograft tumor-bearing mice, nanoCRISPR treatment achieves remarkable anti-tumor efficacy (∼83%) and significant survival benefits with minimal toxicity. This strategy presents a promising paradigm emphasizing transcellular penetration to enhance the effectiveness of CRISPR-based antitumor therapeutics.

Mesangial cell proliferation is an early pathological indicator of diabetic nephropathy (DN). Growing evidence highlights the pivotal role of paired-related homeobox 1 (Prrx1), a key regulator of cellular proliferation and tissue differentiation, in various disease pathogenesis. Notably, Prrx1 is highly expressed in mesangial cells under DN conditions. Both in vitro and in vivo studies have demonstrated that Prrx1 overexpression promotes mesangial cell proliferation and contributes to renal fibrosis in db/m mice. Conversely, Prrx1 knockdown markedly suppresses hyperglycemia-induced mesangial cell proliferation and mitigates renal fibrosis in db/db mice. Mechanistically, Prrx1 directly interacts with the Yes-associated protein 1 (YAP) promoter, leading to the upregulation of YAP expression. This upregulation promotes mesangial cell proliferation and exacerbates renal fibrosis. These findings emphasize the crucial role of Prrx1 upregulation in high glucose-induced mesangial cell proliferation, ultimately leading to renal fibrosis in DN. Therefore, targeting Prrx1 to downregulate its expression presents a promising therapeutic strategy for treating renal fibrosis associated with DN.

Objective:To evaluate the effect of coronary microvascular dysfunction (CMD) on left atrial and ventricular function in patients with ischemia and non-obstructive coronary artery disease (INOCA) under the drug stress of regadenoson by speckle tracking imaging and left ventricular pressure-strain ring ultrasound technology.Methods:A total of 43 patients with INOCA who were admitted to the Department of Cardiology of the First Bethune Hospital of Jilin University from May 2022 to October 2023 were prospectively enrolled, and drug stress tests were performed. The coronary flow velocity reserve (CFVR) values were obtained by transthoracic Doppler echocardiography, and the INOCA patients with CFVR<2.0 were assigned to the CMD group ( n=24), and those with CFVR≥2.0 were assigned to the contrast group (CON group, n=19), and 20 healthy people without chest pain matched by clinical data were selected as the negative group (NEG group). The differences in general clinical data, routine echocardiography before and after stress, left atrial strain, and myocardial work parameters were compared between the groups. The correlation analysis of intra-group parameters was performed, and then the ROC curve was used to evaluate the diagnostic value of ultrasound parameters for INOCA. Results:Compared with the CON group and the NEG group, the ratio of early diastoic velocity E peak of mitral value orifice to the late diastoic velocity A peak(E/A) decreased and the ratio of early diastoic velocity E peak of mitral valve orifice to the early diastoic velocity e′ of the mitral valve annulus(E/e′) increased in the CMD group, and the differences were statistically signnificant (all P<0.05).There were statistically significant differences in left atrial strain parameters including left atrial strain reservoir (LASr), left atrial conduit strain (LAScd), left atrial contraction strain (LASct) between CON group and CMD group before and after stress (all P<0.05).However, there were no statistically significant differences between CON group and CMD group in myocardial work parameters including global longitudinal strain (GLS), peak strain dispersion (PSD), global work index (GWI), global constructive work (GCW), global wasted work (GWW), global work efficiency (GWE) at rest (all P>0.05), and there were significant differences only after stress (all P<0.05). E/e′ was negatively correlated with LASr and LAScd in the CMD group and CON group ( rs=-0.36, r=-0.31; all P<0.05), GLS was positively correlated with GWI, GCW, GWE( r=0.81, 0.61, 0.37; all P<0.05). GLS was positively correlated with GWI, GCW and GWE at stress state( r=0.66, 0.51, 0.52; all P<0.05), and negatively correlated with GWW ( rs=-0.39, P<0.05). PSD was positively correlated with GWW ( rs=0.30, P<0.05), and negatively correlated with GWI, GCW and GWE ( r=-0.46, -0.40, -0.38; all P<0.05). Univariate regression analysis showed that left atrial strain and myocardial work had good predictive values for CMD, and the predictive values of rest LASr and stress GLS were higher, with AUC values of 0.927 and 0.882, respectively. Conclusions:In patients with INOCA and CMD, the left atrial strain capacity decreases at both rest and stress state, and the myocardial work capacity decreases only under the stress. The changes in parameters of left atrial strain and myocardial work provide new ultrasound parameters and predictors for clinical evaluation of CMD.

Objective:To investigate the influence of different test conditions on the particle size distribution of elemene emulsion injection measured by laser light scattering method.Methods:The method of particle size deter-mination of elemene emulsion injection was explored.The test conditions were as follows:shading rate 5％-10％,absorption rate 0.000 1-0.1,refractive index1.52-1.54,mixing speed 800-1 600 r·min-1,balance time 0.5-1 min.After the methodology verification,3 batches of elemene emulsion injection were taken and deter-mined on the Bettersize 2600 laser particle size distribution instrument(wet method).Results:Measurement results of 3 batches of samples:d(0.1),d(0.5),d(0.9),d(0.99)were 252,251,213 nm;354,351,316 nm;543,532,476 nm;979,953,887 nm.Conclusion:This test method can be used to determine the particle size distribution of elemene emulsion injection with good reproducibility.

Fibroblast growth factors (FGF) are a group of structurally related polypeptides which constitute an elaborate signaling system with their receptors. Evidence accumulated in the years suggests that the FGF family plays a key role in the repair of central nervous system injury. The main protective mechanisms include activating the expression of PI3K-Akt, peroxisome proliferator-activated receptor (PPARγ) and other signals; inhibiting NF-κB-mediated inflammatory response, oxidative stress and apoptosis; regulating neuronal differentiation and neuronal excitability as well as participating in protection of neurovascular units and nerve function repair. This paper comprehensively summarizes the latest research progress in FGF signaling related to diseases of the central nervous system such as cerebral infarction, cerebral hemorrhage, traumatic brain injury, Alzheimer's disease, Parkinson's disease, epilepsy and depression, aiming to provide scientific basis and reference for the development of innovative FGF drugs for the prevention and treatment of neurological diseases.
Humans ; Fibroblast Growth Factors ; Phosphatidylinositol 3-Kinases/metabolism* ; Central Nervous System/metabolism* ; Signal Transduction/physiology* ; Alzheimer Disease

Objective:To investigate the role and diagnostic value of miRNA-205 in chronic kidney disease (CKD) patients with vascular calcification.Methods:It was divided into in vitro cell experiment and retrospective cohort study. In vitro experiments were conducted by using rat thoracic aortic smooth muscle cells. Alizarin red staining and calcium content detection were used to detect the calcification of vascular smooth muscle cells (VSMCs). Alkaline phosphatase (ALP) test kit was used to measure ALP activity. Western blotting was used to detect the protein expression levels of osteogenic transcription factors runt-related transcription factor 2 (Runx2), α smooth muscle actin (α-SMA) and smooth muscle-22α (SM-22α) in VSMCs. qRT-PCR was used to detect miRNA-205 and Runx2 expression levels. The double luciferase reporter gene assay was used to verify the targeted relationship between miRNA-205 and Runx2. The non-dialysis patients with CKD 3-5 stage from June 2020 to January 2021 in the Department of Nephrology of Fourth Hospital, Hebei Medical University were selected. According to coronary artery calcium score (CACs), the patients were divided into non-calcification group (CACs=0), mild-moderate calcification group (0 400). Spearman correlation analysis was used to analyze the correlation between miRNA-205 and Runx2 and vascular calcification. Logistic regression model and receiver operating characteristic (ROC) curve analysis were used to analyze the ability of miRNA-205 to predict the vascular calcification in patients with CKD. Results:(1)Compared with the control group, calcium nodules were more, and the calcium content, ALP activity and Runx2 protein level were higher, and the expression levels of miRNA-205, α-SMA and SM-22α were significantly lower in high phosphorus group (all P<0.05). Overexpression of miRNA-205 significantly reduced the calcification of VSMCs and Runx2 protein level, and increased the protein levels of α-SMA and SM-22α (all P<0.05). miRNA-205-5p reduced the activity of luciferase in the wild-type Runx2-3'-end non-coding region plasmid. (2) Eighty CKD patients were enrolled, with age of (57.50±14.93) years old and 49 males (61.3%). The results of comparison of miRNA-205 and Runx2 expression levels in non-calcification group ( n=26), mild- moderate calcification group ( n=30) and severe calcification group ( n=24) showed that, the higher degree of calcification, the lower miRNA-205 expression level and the higher Runx2 mRNA expression level (all P<0.05). miRNA-205 was negatively correlated with CACs ( r=-0.50, P<0.01) and Runx2 was positively correlated with CACs ( r=0.55, P<0.01). Multivariate logistic regression analysis results suggested that miRNA-205 ( OR=0.451, 95% CI 0.122-0.873) was an independent influencing factor of vascular calcification in CKD patients. The area under the ROC curve of miRNA-205 and miRNA-205 combined with Runx2 for predicting vascular calcification were 0.796 (95% CI 0.697-0.859) and 0.924 (95% CI 0.866-0.982), respectively. Conclusions:miRNA-205 inhibits vascular calcification by targeting Runx2 to negatively regulate osteogenetic phenotype transformation of VSMCs and is expected to be an early diagnostic marker of vascular calcification in CKD patients.

Objective:To systematically analyze and evaluate the predictive performance of 5 assessment scales (Munro Pressure Ulcer Risk Assessment Scale, Norton Scale, Braden Scale, Scott Triggers Scale and Waterlow Scale) for adult intraoperatively acquired pressure injury (IAPI), so as to provide reference for the selection of appropriate assessment scales.Methods:This study is a Meta-analysis. Research on the adult IAPI assessment scale was retrieved through computers in databases such as VIP, WanFang Data, China National Knowledge Infrastructure, China Biology Medicine disc, PubMed, Embase, Web of Science and Cochrane Library. The search time limit was from the establishment of the database to December 2, 2021.Two researchers independently screened the article, conducted quality evaluation and data extraction on the included article, and conducted descriptive analysis on the sensitivity and specificity of each scale. The researchers used RevMan 5.3 software to create a receiver operating characteristic (ROC) curve and calculate the area under the curve.Results:A total of 13 articles were included, with a total of 5 640 cases, and the incidence of IAPI was 8.09% (456/5 640). A Meta-analysis was conducted after merging 8 articles on the Braden scale. After merging, it was found that the sensitivity of the scale was 0.76 [95% confidence interval ( CI) (0.72, 0.81) ], the specificity was 0.53 [95% CI (0.51, 0.54) ], and the area under the ROC curve was 0.70. After a combined analysis of 7 articles on the Munro Pressure Ulcer Risk Assessment Scale, it was found that the sensitivity of the scale was 0.81 [95% CI (0.75, 0.86) ], the specificity was 0.79 [95% CI (0.77, 0.81) ], and the area under the ROC curve was 0.84. After a combined analysis of 4 articles on the Waterlow scale, it was found that the sensitivity of the scale was 0.78 [95% CI (0.69, 0.86) ], the specificity was 0.60 [95% CI (0.58, 0.62) ], and the area under the ROC curve was 0.72. After a combined analysis of 3 articles on the Norton scale, it was found that the sensitivity of the scale was 0.67 [95% CI (0.56, 0.77) ], the specificity was 0.69 [95% CI (0.67, 0.71) ], and the area under the ROC curve was 0.74. After a combined analysis of 3 articles on the Scott Triggers Scale, it was found that the sensitivity of the scale was 0.80 [95% CI (0.68, 0.89) ], the specificity was 0.53 [95% CI (0.50, 0.56) ], and the area under the ROC curve was 0.67. Conclusions:As a special scale for surgical patients, the Munro scale has superior sensitivity, specificity, and area under the ROC curve to other evaluation scales. It is recommended that Operating Room nurses prioritize the Munro scale for assessing the risk of IAPI in adult surgical patients.

Objective:To explore the effect of Toll-like receptor 9 (TLR9) signaling pathway activation on the transcriptome in the renal tubular cells.Methods:Mouse primary renal tubular epithelial cells were extracted and cultured. When the degree of cell fusion reached 80%, they were divided into two groups, which were added with 10 μL phosphate buffered saline (PBS, PBS control group) and TLR9 activator cytosine phosphate guanidine oligodeoxynucleotide (CpG-ODN) with a final concentration of 5 μmol/L (CpG-ODN treatment group). The RNA sequencing was performed on the Illumina platform after extraction. DEGseq software was used to analyze the differential expression of genes between the two groups. Goatools and KOBAS online software were used to analyze the differential genes involved signal pathways. Homer software was used to predict transcription factors.Results:Compared with the PBS control group, there were a total of 584 differentially expressed genes in the CpG-ODN treatment group, of which 102 were up-regulated and 482 were down-regulated. The most significantly enriched gene ontology (GO) terms of differentially expressed genes included response to interferon-β, defense response to virus and other inflammatory pathway. The most significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways included 2'-5'-oligoadenylate synthase activity, regulation of ribonuclease activity, negative regulation of virus life cycle, cellular response to interferon-βand defense response to protozoan. The results of transcription factor prediction showed that interferon regulatory factor 3 (IRF3) was the most significantly enriched transcription factor in the promoter sequence of differential genes; the most significant transcription factor downstream of TLR9 was IRF3, and other predicted transcription factors such as transcription factor 21 (TCF21), zinc finger protein 135 (ZNF135), and PR domain containing 4 (PRDM4) might be new candidates for TLR9 signaling pathway.Conclusion:CpG-ODN activates TLR9 signaling pathway, and primary renal tubular epithelial cells can directly respond to CpG-ODN stimulation and undergo transcriptome changes, which provides a basis for further research on the molecular mechanism of TLR9 pathway in sepsis induced acute kidney injury.

Objective To analyze the risk factors of in-hospital mortality in patients with intracerebral hemorrhage (ICH) in the intensive care unit.Methods Patients with intracerebral hemorrhage were retrospectively collected from January 2013 to January 2018 in the Department of Intensive Care Unit,Zhongnan Hospital of Wuhan University.Patients were excluded aged less than 18 years,pregnant women,the onset time of more than 7 days,the length of hospital stay of less than 48 hours,lack of renal function outcomes within 24 hours after admission,the glomerular filtration rate (eGFR) of lower than 15 mL/(min.1.73 m2),the history of chronic kidney disease,regular dialysis and renal transplantation,and incomplete data.Clinical data were collected from baseline characteristics,past history,and laboratory examination.The included patients were divided into the in-hospital non-survival group and the survival group.SPSS 20.0 software as used for statistical analysis,the binary Logistic regression analysis was performed to evaluate risk factors of in-hospital mortality with intracerebral hemorrhage,the prognosis was assessed by receiver operating characteristic (ROC) curve and survival curve (Kaplan-Meier).A P＜0.05 was considered statistically significant.Results In this single-center retrospective study,a total of 300 patients were enrolled,including 96 patients in the hospital nonsurvival group and 204 patients in the survival group.The incidence of in-hospital death in patients with intracerebral hemorrhage in ICU was 32％.Multivariate analysis demonstrated that the risk factors of inhospital mortality were lower GCS score (OR=0.629,95％CI:0.523-0.757,P＜0.01),higher APACHE Ⅱ score (OR=1.590,95％CI:1.369-1.847,P＜0.01),elevated leukocytes (OR=1.082,95％CI:1.028-1.139,P=0.002) and the incidence of acute kidney injury (AKI) (OR=6.978,95％CI:3.381-14.405,P＜0.01).The ROC curve demonstrated that the area under curve (AUC) of APACHE Ⅱ score was the largest with a sensitivity and specificity of 73.96％ and 75.98％,respectively,which can better predict the mortality of patients with cerebral hemorrhage.Kaplan-Meier survival curve showed that in-hospital survival rate of non-AKI patients were higher than that of AKI patients (P＜0.01).Conclusions Lower GCS score,higher APACHE Ⅱ score,elevated white blood cells and AKI are risk factors for predicting in-hospital mortality in patients with intracerebral hemorrhage in the ICU.Therefore,early identification and treatment should be adopted in these high-risk populations.

Objective: To evaluate the performance of Vitek MS, an automated mass spectrometry microbial identification system, in identification of clinical filamentous fungi isolates, which provide a new insight into filamentous fungi identification strategy. Methods: Twenty-five isolates of filamentous fungi genetically confirmed by molecular sequencing method were inoculated in different culture conditions to analyze the potential influences of culture conditions on the identification of filamentous fungi with Vitek MS. Further evaluation of the performance of Vitek MS in filamentous fungi identification was carried out with 207 clinical isolates of filamentous fungi and the comparison of the results with those of morphology and sequencing analysis. Results: The performance of Vitek MS in filamentous fungi identification was not significantly influenced by culture conditions. Vitek MS successfully identified 87.9% (182/207) of all tested filamentous fungi. Lacking of reference MS spectra in Vitek database was the main reason that the other isolates could not be identified. Vitek MS was superior to the traditional morphological method in identification of filamentous fungi especially non-Aspergillus molds at species level. Conclusion Vitek MS is an efficient approach for clinical filamentous fungi identification. Continuous enrichment of the species coverage in database will be of great importance for improving the performance of Vitek MS in identification of filamentous fungi.