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.

Objective:To investigate the association of exposure to PM 2.5 and its constituents during pregnancy and fetal growth and to further identify critical windows of exposure for fetal growth. Methods:We included 4 089 mother-child pairs from the Jiangsu Birth Cohort Study between January 2016 and October 2019. Data of general characteristics, clinical information, daily average PM 2.5 exposure, and its constituents during pregnancy were collected. Fetal growth parameters, including head circumference (HC), abdominal circumference (AC), and femur length (FL), were measured by ultrasound after 20 weeks of gestation, and then estimated fetal weight (EFW) was calculated. Generalized linear mixed models were adopted to examine the associations of prenatal exposure to PM 2.5 and its constituents with fetal growth. Distributed lag nonlinear models were used to identify critical exposure windows for each outcome. Results:A 10 μg/m 3 increase in PM 2.5 exposure during pregnancy was associated with a decrease of 0.025 ( β=-0.025, 95% CI: -0.048- -0.001) in HC Z-score, 0.026 ( β=-0.026, 95% CI: -0.049- -0.003) in AC Z-score, and 0.028 ( β=-0.028, 95% CI:-0.052--0.004) in EFW Z-score, along with an increased risk of 8.5% ( RR=1.085, 95% CI: 1.010-1.165) and 13.5% ( RR=1.135, 95% CI: 1.016-1.268) for undergrowth of HC and EFW, respectively. Regarding PM 2.5 constituents, prenatal exposure to black carbon, organic matter, nitrate, sulfate (SO 42-) and ammonium consistently correlated with decreased HC Z-score. SO 42- exposure was also associated with decreased FL Z-scores. In addition, we found that gestational weeks 2-5 were critical windows for HC, weeks 4-13 and 19-40 for AC, weeks 4-13 and 23-37 for FL, and weeks 4-12 and 20-40 for EFW. Conclusions:Our findings demonstrated that exposure to PM 2.5 and its constituents during pregnancy could adversely affect fetal growth and the critical windows for different fetal growth parameters are not completely consistent.

Recent trends suggest that Chinese herbal medicine formulas(CHM formulas)are promising treatments for complex diseases.To characterize the precise syndromes,precise diseases and precise targets of the precise targets between complex diseases and CHM formulas,we developed an artificial intelligence-based quantitative predictive algorithm(DeepTCM).DeepTCM has gone through multilevel model cali-bration and validation against a comprehensive set of herb and disease data so that it accurately captures the complex cellular signaling,molecular and theoretical levels of traditional Chinese medicine(TCM).As an example,our model simulated the optimal CHM formulas for the treatment of coronary heart disease(CHD)with depression,and through model sensitivity analysis,we calculated the balanced scoring of the formulas.Furthermore,we constructed a biological knowledge graph representing interactions by associating herb-target and gene-disease interactions.Finally,we experimentally confirmed the thera-peutic effect and pharmacological mechanism of a novel model-predicted intervention in humans and mice.This novel multiscale model opened up a new avenue to combine"disease syndrome"and"macro micro"system modeling to facilitate translational research in CHM formulas.

Objective To investigate the expression of Bone morphogenetic protein 4(BMP4)in human endometrial stromal cells(hESCs)of patients with recurrent spontaneous abortion(RSA)and the regulation of Forkhead transcription factor 1(FOXO1)by BMP4.Methods The decidua tissues of RSA patients and Healthy control(HC)patients in People's Hospital of Wuhan University were collected,and the expression of BMP4 was analyzed by immunohistochemistry and RT-qPCR.hESC was used in vitro cell experiments,lentivirus was transfected to knock down the expression of BMP4 and establish the cell model of RSA.After decidualization,CCK-8 as-say,inverted microscope,and Western blot assay were used to detect the cell proliferation,morphology,and expression levels of decidual-ization marker genes.To verify whether BMP4 knockdown affects FOXO1 expression,a FOXO1 inhibitor(AS1842856)was used to per-form a response experiment,and the upstream and downstream relationship between FOXO1 and BMP4 was analyzed by CCK-8 experi-ment,inverted microscope,and Western blot.In animal experiments,intraperitoneal injection of LPS-induced abortion in pregnant mice,and the expression of BMP4 in decidua were detected to explore the relationship between BMP4 and abortion in mice.Results Compared with the HC group,the expression of BMP4 in the deciduous tissues of RSA patients was significantly decreased.After BMP4 was knocked down,the decidualization was significantly abnormal,which showed that the proliferation rate increased,the cell morphologi-cal transformation was not obvious,and the expression of decidualization marker genes was down-regulated.FOXO1 was the downstream regulatory molecule of BMP4,knocking down BMP4 could inhibit the expression of FOXO1,and BMP4 could regulate the decidualization of hESC through FOXO1.In animal models,the expression of BMP4 in decidua and placental weight of aborted pregnant mice were signif-icantly reduced.Conclusion The expression of BMP4 was significantly down-regulated in decidual tissues of RSA patients and aborted mice.In cell experiments,BMP4 can promote decidualization of hESC by regulating the expression of FOXO1.Abnormally low expression of BMP4 may be one of the reasons for decidualization defects in RSA patients.

Objective:To investigate the clinicopathological features, molecular pathology characteristics, and prognosis of non-small cell lung carcinoma (NSCLC) exhibiting co-expression of p40 and thyroid transcription factor1 (TTF1).Methods:Clinical and pathological data of six NSCLC cases with co-expression of p40 and TTF1 diagnosed at the First People′s Hospital of Xiaoshan District, Hangzhou, China from January 2016 to December 2023 were collected. Relevant literature was also reviewed.Results:NSCLC with co-expression of p40 and TTF1 commonly occurred in male smokers and had been in stage Ⅲ-Ⅳ when diagnosis. Microscopic examination revealed that the tumor cells were arranged in solid nests and sheets with marked atypia and visible mitotic figures. There was no prominent evidence of keratinization or glandular formation. The tumor cells diffusely co-expressed p40 and TTF1, exhibiting a dual immunophenotype characteristic of both squamous cell carcinoma and adenocarcinoma. Molecular testing of four NSCLC co-expressing p40 and TTF1 revealed the presence of common EGFR mutations, as well as mutations of NRAS (mutation rate of 2.09%), EML4-ALK (mutation rate of 24.77%), and PIK3CA (exon 10 c.1658 G>C p.S553T, mutation rate of 4.32%). All six tumors were poorly differentiated, highly invasive, and associated with poor prognosis. Four of the six patients experienced widespread metastasis and died within 7 to 30 months after the diagnosis or initial treatment.Conclusions:NSCLC with co-expression of p40 and TTF1 exhibits distinct clinicopathological features, immunophenotypes, molecular alterations, and clinical outcomes, characterized by rapid progression and poor prognosis. Pathologists should be vigilant in recognizing this entity to avoid misdiagnosis and missed diagnosis.

Objective We aimed to investigate the relationship between microscopic pattern of blood stasis and renal pathological grade and related physical and chemical indexes in children with Henoch-Sch?nlein purpura nephritis(HSPN).Methods We conducted a retrospective analysis of 800 HSPN children from the medical records of the First Affiliated Hospital of Henan University of Chinese Medicine.Laboratory indicators(blood routine test,urine routine test,coagulation test,liver function)and renal pathological indicators of them were collected.According to the severity of renal pathological microscopic lesions,the microscopic pattern of blood stasis was divided into three types,including choroidal discord,dead blood coagulation and intracarenal disease accumulation.The classification of renal microscopic pattern of blood stasis and the correlation between laboratory indexes and renal pathological index were analyzed by Spearman grade correlation and binary Logistic regression analysis.Results(ⅰ)There was no statistical difference of the distribution of the renal microscopic pattern of blood stasis in the different traditional Chinese medicine patterns.(ⅱ)There were significant differences in the contents or the grade of albumin and fibrinogen in the HSPN children with different microscopic pattern of blood stasis(all P＜0.05).(ⅲ)The maximum area under the receiver operating characteristic(ROC)curve between fibrinogen and intracarenal disease accumulation was 0.594(95％CI from 0.540 to 0.633,P＜0.001);sensitivity was 0.447,specificity was 0.725;the best threshold on the ROC curve of 0.172 was 3.755 g/L.(ⅳ)There were positive correlations between the content of fibrinogen,ISKDC grade and Bohle A grade respectively with the scores of intracarenal disease accumulation type(r=0.176,r=0.315,r=0.656;all P＜0.001).(ⅴ)There were positive correlations between the content of fibrinogen,ISKDC grade and Bohle A grade respectively with the renal microscopic pattern of blood stasis(r=0.157,r=0.377,r=0.429;all P＜0.001).Conclusion The microscopic renal pattern of blood stasis can not only reflect the severity of renal blood stasis,but also reflect the severity and long-term prognosis of renal diseases.Albumin and urinary protein grade can reflect the early stage of the microscopic renal pattern of the blood stasis(choroidal discord).The content of fibrinogen increases with the aggravation of renal microscopic pattern of blood stasis,reflecting the end-stage of HSPN,which has the correlation with the formation and severity of related indexes.Fibrinogen can be used as a laboratory indicator to assist in the diagnosis of irreversible lesionsin the renal pathology of HSPN children.

Objective:To explore the current status of lipid-lowering therapy, the distribution of low-density lipoprotein cholesterol (LDL-C) levels and the risk assessment of cardiovascular events recurrence in patients with coronary heart disease (CHD) complicated by hypertension.Methods:This was a cross-sectional study. Patients with CHD combined with hypertension were hospitalized in the Department of Cardiology, General Hospital of Chinese People′s Liberation Army from August 5, 2008 to July 22, 2018 were included, and were divided into standard group and substandard group according to whether LDL-C reached the standard. Study data were obtained from inpatient coronary angiography records and electronic medical records database of Department of Cardiology, General Hospital of Chinese People′s Liberation Army, who used data from the first diagnosis of CHD. Clinical data of the selected patients were collected. Multivariate logistic regression model was used to analyze the associated factors of whether LDL-C reached the standard in CHD patients with hypertension.Results:A total of 18 800 patients were selected from 31 provinces/autonomous regions/municipalities directly under the central government in China, with Beijing accounting for the largest proportion (5 692 patients (30.28%)), followed by Hebei (3 621 patients (19.26%)), Henan (1 837 patients (9.77%)), and Shandong (1 618 patients (8.61%)). Among the selected patients, 1 493 had LDL-C<1.4 mmol/L (standard group), and 17 307 had LDL-C≥1.4 mmol/L (substandard group). Only 1 493 patients (7.94%) had LDL-C<1.4 mmol/L. There were 4 518 patients (24.03%), 4 366 patients (23.22%), 6 924 patients (36.83%) and 1 499 patients (7.97%) with LDL-C for 1.4-<2.0, 2.0-<2.5, 2.5-<3.8 and≥3.8 mmol/L levels, respectively. 17 855 patients (95.15%) were treated with statins, but only 1 334 patients (7.10%) were treated with statins and ezetimibe. Of the selected patients, 4 986 patients (26.52%) were at low risk, 6 515 patients (34.65%) were at intermediate risk, and 7 299 patients (38.82%) were at high risk. The combined lipid-lowering treatment rates of statin and ezetimibe in the middle-and high-risk patients were 7.43% (484/6 515) and 7.48% (546/7 299), respectively. The results of multivariate logistic regression analysis showed that increasing age, male, diabetes mellitus, stroke, and history of percutaneous coronary intervention (PCI) were positively associated with LDL-C standards in patients with CHD and hypertension, whereas obesity and acute myocardial infarction (AMI) were negatively associated with LDL-C standards (all P<0.01). Conclusions:The rate of achieving the standard LDL-C in patients with CHD combined with hypertension was low in China. Although the majority of patients had received moderate-intensity statin therapy, the proportion of statin-treated patients combined with ezetimibe was extremely low. The proportion of high-risk patients with recurrent cardiovascular events was higher in patients with CHD and hypertension in China, whereas the proportion of such patients receiving statin combined with ezetimibe lipid-lowering therapy was lower. This study also found that increasing age, male, diabetes mellitus, stroke, and history of PCI were positively associated with LDL-C standards, while obesity and AMI were negatively associated with LDL-C standards in patients with CHD and hypertension.

Objective: To explore the relationship between 24 hour movement behavior and physical fitness in college students, so as to provide a reference for physical fitness interventions for college students. Methods: A survey was conducted from September 23 to November 12, 2022, involving 342 college students selected from one comprehensive college among Shanghai City, Hubei Province and Zhejiang Province. Accelerometers were used to measure 24 hour movement behavior, and programs specified in the National Physical Fitness Standards for Students (2014 revision) were used to evaluate physical fitness. Compositional linear regression was used to analyze the association between 24 hour movement behavior and physical fitness. The effects of compositional isotemporal substitution of 24 hour movement behavior on physical fitness were also examined. Results: The geometric means for sleep, sedentary behavior, light physical activity (LPA), and moderate to vigorous physical activity (MVPA) were 515.89, 678.88 , 196.30, 48.92 min/d, respectively, and the comprehensive physical fitness score was (73.09±8.55). The proportion of participants who passed the fitness test was 72.51%. Overall, 20.76% had excellent or good scores, whereas 6.73% failed. Compositional linear regression indicated that physical fitness was significantly positively associated with MVPA ( β =2.55) and LPA ( β = 5.88 )( P <0.05), but no significantly associated with sleep ( β =-5.18) and sedentary behavior ( β =-3.24)( P >0.05). Isotemporal substitution indicated that reallocation of 15 minutes from sleep and sedentary behavior to MVPA resulted in a 0.71 and 0.64 point increase in physical fitness, respectively. Similarly, reallocating the same duration to LPA led to a 0.50 and 0.43 point increase in physical fitness, respectively. Dose response analysis revealed that the mutual substitution of LPA with sedentary behavior and sleep had symmetry, whereas the mutual substitution of MVPA with sedentary behavior and sleep had asymmetry. The effectiveness of replacing sleep or sedentary behavior with MVPA in improving physical fitness decreased with increasing allocation time, whereas the adverse effects of replacing MVPA with sleep or sedentary behavior increased with increasing allocation time. Conclusion In the context of 24 hour movement behavior, increasing the time spent on LPA and MVPA is beneficial for physical fitness among college students. It should be recommend that college students actively transition from sedentary behavior to engaging in physical activity, with a particular emphasis on incorporating MVPA.

Objective:To investigate the efficacy of radiofrequency ablation combined with ozone therapy under CT guidance in the treatment of lumbar disc herniation.Methods:A total of 93 patients with lumbar disc herniation who were admitted to The First Hospital of Jiaxing from January 2019 to May 2022 were included in this study. They were randomly divided into an observation group ( n = 47) and a control group ( n = 46). The control group was treated with radiofrequency ablation and the observation group was treated with radiofrequency ablation combined with ozone therapy. Efficacy was compared between the two groups at 3 months after surgery. The pain was compared between the two groups before and 7 days and 3 months after surgery. Inflammatory factors were compared between the two groups before and 7 days after surgery. The lumbar spine function was compared between the two groups before and 3 months after surgery. Results:At 3 months after surgery, the excellent and good rate in the observation group was significantly higher than that in the control group [89.36% (42/47) vs. 71.74% (33/47), χ2 = 4.63, P < 0.05). At 7 days and 3 months after surgery, Visual Analogue Scale scores in the observation group were (2.91 ± 0.54) points and (1.32 ± 0.31) points, respectively, which were significantly lower than (3.76 ± 0.62) points and (2.08 ± 0.47) points in the control group ( t = 7.06, 9.22, both P < 0.001). At 7 days after surgery, serum interleukin-1β, interleukin-6, and tumor necrosis factor-α in the observation group were (0.24 ± 0.05) μg/L, (18.49 ± 3.47) ng/L, and (97.94 ± 17.43) ng/L, respectively, which were significantly lower than (0.37 ± 0.09) μg/L, (24.31 ± 4.12) ng/L, and (148.87 ± 20.13) ng/L, respectively in the control group ( t = 8.63, 7.37, 13.05, all P < 0.05). At 3 months after surgery, the Japanese Orthopedic Association score in the observation group was significantly higher than that in the control group [(25.68 ± 2.28) points vs. (21.17 ± 3.24) points, t = -7.78, P < 0.001], and the Oswestry Disability Index in the observation group was significantly lower than that in the control group [(9.84 ± 1.43) points vs. (13.46 ± 2.18) points, t = 9.49, P < 0.001]. Conclusion:Radiofrequency ablation combined with ozone therapy under CT guidance is highly effective on lumbar disc herniation. The combined therapy can reduce pain and inflammatory reactions in patients and improve lumbar function.

Objective To investigate the syndrome differentiation characteristics of children with atopic dermatitis(AD)and the immune imbalance status in children with different syndrome types of AD.Methods A total of 159 AD children and 100 normal control children were enrolled.The peripheral blood eosinophil(Eo)count was measured by impedance method,total serum immunoglobulin E(IgE)by immunoturbidimetric assay,and interferon-gamma(IFN-γ),interleukin-4(IL-4),interleukin-5(IL-5)and interleukin-17(IL-17)were measured by multiple microspheres flow immunofluorescence assay.Results Among 159 AD children,syndrome of heart-fire and spleen-deficiency was most commom,accounting for 38.4%,followed by syndrome of blood-deficiency and wind-dryness(22.0%),syndrome of heat accumulation in heart and spleen(20.1%)and syndrome of spleen-deficiency and dampness-accumulation(19.5%).Compared with normal control group,there was no significant difference in serum IFN-γ level among different syndrome types of AD.The levels of peripheral blood Eo,serum total IgE,IL-4 and IL-17 in AD with heart-fire and spleen-deficiency syndrome were significantly increased(P<0.05).The levels of peripheral blood Eo,IL-4,IL-5 and IL-17 in AD with blood-deficiency and wind-dryness syndrome were significantly increased(P<0.05).The levels of IL-4,IL-5 and IL-17 in AD with heat accumulation in heart and spleen syndrome were significantly increased(P<0.05).The levels of peripheral blood Eo and serum IL-4 in AD with spleen-deficiency and dampness-accumulation syndrome were significantly increased(P<0.05).Conclusion Heart-fire and spleen-deficiency syndrome is the most common type in children with AD,however,the main type under 3 years old is heat accumulation in heart and spleen syndrome.Th2/Th17 immune imbalance are the main pathogenesis in heart-fire and spleen-deficiency syndrome,blood-deficiency and wind-dryness syndrome and heat accumulation in heart and spleen syndrome,and Th2 immune imbalance is the main pathogenesis of spleen-deficiency and dampness-accumulation syndrome.