BACKGROUND: Electroacupuncture (EA) treatment is efficacious in patients with respiratory disorders, although the mechanisms of its action in lung-function protection are poorly understood. This study aimed to explore the neuroanatomical mechanisms of EA stimulation at the BL13 acupoint (Feishu, EA-BL13) improvement in asthma. METHODS: Allergic asthma was induced by intranasal 2.0% ovalbumin (OVA) instillation combined with intraperitoneal injection of the 10.0% OVA. The levels of interleukin (IL)-4 and IL-5 were detected by enzyme-linked immunosorbent assay. Hematoxylin and eosin and periodic acid-schiff stain were used to evaluate inflammatory cell infiltration and mucus secretion. Cellular oncogene fos induction in neurons after EA stimulation was detected by immunofluorescent staining. The messenger RNA expression levels of adrenergic receptors were quantified with real-time polymerase chain reaction. RESULTS: EA improved airway inflammation and mucus secretion mainly by activating somatosensory-sympathetic pathways ( P <0.001). Briefly, the intermediolateral (IML) nuclei of the spinal cord received signals from somatic EA stimulation and then delivered the information via the sympathetic trunk to the lung. Excited sympathetic nerve endings in lung tissue released large amounts of catecholamines that specifically activated the β2 adrenergic receptor (β2AR) on T cells ( P <0.01) and further decreased the levels of IL-4 and IL-5 ( P <0.001) through the cyclic adenosine monophosphate/protein kinase A signaling pathway. CONCLUSION This study provided a new explanation and clinical basis for the use of EA-BL13 as a treatment for allergic asthma in both the attack and remission stages and other respiratory disorders related to airway inflammation.
Electroacupuncture/methods* ; Animals ; Asthma/immunology* ; Rats ; Rats, Sprague-Dawley ; Male ; Inflammation/therapy* ; Interleukin-4/metabolism* ; Interleukin-5/metabolism*

Objective To evaluate the renoprotective effects of zinc(Zn)supplementation in diabetes kidney disease(DKD)and to explore its impact on the nuclear factor erythroid 2-related factor 2(Nrf2)signaling pathway.Methods A total of 12 male OVE26 mice(spontaneous type 1 diabetes mellitus mice)aged 3 months and weighing approximately 24-27 g were selected and randomly assigned to a diabetes mellitus(DM)group and a zinc-treated DM(DM/Zn)group(n=6 each).In addition,12 age-matched male FVB mice weighing approximately 27-30 g were selected and randomly assigned to a non-diabetic control(Ctrl)group and a zinc-treated(Zn)group(n=6 each).Mice in the DM/Zn and Zn groups were given zinc supplementation for 3 months,with each mouse receiving 5 mg/kg of zinc sulfate by gavage every other day.Mice in the DM and Ctrl groups were given the same volume of normal saline.At the end of the experiment,the albumin-to-creatinine ratio(ACR)in urine was used as an indicator to evaluate renal function.Sirius red staining was performed to assess renal fibrosis in each group of mice.Western blotting was performed to determine the expression of fibrotic growth factors,including connective tissue growth factor(CTGF)and transforming growth factor-β1(TGF-β1),in renal tissue,and the protein expression of Nrf2,an antioxidant substance,and the protein expression levels of its downstream targets,including NAD(P)H quinone dehydrogenase 1(NQO1),heme oxygenase 1(HO-1),superoxide dismutase(SOD)-1,SOD-2,and catalase(CAT).Results 1)Compared to the Ctrl group,the urinary protein secretion levels of mice in the DM group exhibited progressive increase.After 3 months of zinc supplementation treatment,the urinary protein secretion levels of mice in the DM/Zn group decreased Compared to that of mice in the DM group,and the difference was statistically significant(P<0.05).2)Compared to that in the Ctrl group,the collagen deposition in the renal tissues of mice in the DM group increased,and the difference was statistically significant(P<0.05),while no obvious change was observed in mice in the DM/Zn group.Compared to the Ctrl group,mice in the DM group exhibited increased expression levels of CTGF and TGF-β1 in the renal tissues,but the expression levels decreased after zinc supplementation treatment,with the differences being statistically significant(P<0.05).3)Compared to that of the Ctrl group,the expression level of Nrf2 in the renal tissues of mice in the Zn and DM groups increased,and the level of Nrf2 in the renal tissues of mice in the DM/Zn group showed a further increase,with the differences being statistically significant(P<0.05).4)Compared to those of the Ctrl group,the protein expression levels of Nrf2 downstream target genes,including NQO1 and HO-1,in the renal tissues of mice in the Zn group increased,and the levels of NQO1 and HO-1 in the renal tissues of mice in the DM/Zn group showed a further increase,with the differences being statistically significant(P<0.05).Compared to those of the mice in the Ctrl group,the protein expressions of Nrf2 downstream target genes,including SOD-1,SOD-2,and CAT of in the renal tissues of the mice in the Zn group increased,while the expression levels of SOD-1,SOD-2,and CAT in the renal tissues of the mice in the DM group decreased,with the differences being statistically significant(P<0.05).Zn supplementation could completely inhibit these changes(P<0.05).Conclusions Zn supplementation has therapeutic effects on DKD and mitigates T1DM-induced renal dysfunction and oxidative injury in mice,which may be associated with the activation of the Nrf2 antioxidant signaling pathway.

Objective To investigate the effects of DNA demethylation drugs combined with histone deacetylase in-hibitors on fragile X mental retardation 1 neighbor protein (FMR1NB) expression and its promoter methylation in human oral cancer cells and try to find a strategy of weakening the heterogeneity of FMR1NB expression.Methods Human oral cancer cell lines Cal27 and SCC-9 were treated with decitabine (DAC) , an inhibitor of DNA meth-yltransferase, combined with trichostatin A (TSA) and valproic acid (VPA), inhibitors of histone deacetylase.Then reverse transcription-polymerase chain reaction (RT-PCR) , quantitative real-time PCR (qRT-PCR) and Western blot were used to detect the expression of FMR1 NB and pyrosequencing was used to detect the methylation of FMR1NB promoter.Results Compared with the blank control group, DAC and its combination with TSA and VPA significantly induced the expression of FMR1NB mRNA and protein in Cal27 and SCC-9 cells.Compared with DAC alone group, FMR1NB mRNA expression of each DAC-combined drug groups significantly increased, but FMR1NB protein did not significantly change in Cal27 cells; for SCC-9 cells, except for DAC+TSA group, the mRNA and protein levels of FMR1NB significantly increased in all other groups.In addition, there was no signifi-cant difference in the expression of FMR1 NB mRNA and protein between the three-combined drugs group and two-combined drugs groups.Further methylation assay showed that the methylation level of the overall FMR1NB promot-er and its each CpG site measured were reduced to varying degrees in all treatment groups except for three-combina-tion drug group of SCC-9.Conclusion DAC and its combination with TSA and VPA can enhance the expression of FMR1NB by mediating the demethylation of FMR1NB promoter, wherein the enhanced expression effect of the com-bination of the two drugs is stronger, suggesting that they have the potential to weaken the heterogeneity of FMR1NB expression and improve the immunotherapy effect of oral cancer.

Urine is produced from the urinary system, and urinary cell-free DNA (cfDNA) carries genomic DNA directly secreted from urinary system.Urine samples are non-invasive, unlimited in quantity and easy to obtain, making urinary cfDNA a promising biomarker for urologic diseases.This article reviews the progress of clinical application of urinary cfDNA in urologic diseases.

Objective:To evaluate the correlation between vascular hyperintensity of magnetic resonance fluid-attenuated inversion recovery (FLAIR) sequence(FVH) and related parameters of magnetic resonance perfusion weighted imaging (MR-PWI) in patients with middle cerebral artery stenosis cerebral infarction, and to explore the hemodynamic factors related to FVH and the effect of FVH on the short-term clinical prognosis of patients.Methods:A total of 116 patients with middle cerebral artery stenosis cerebral infarction in the Department of Neurology, Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University from January 2020 to December 2022 were collected.According to the diagnostic criteria of FVH, they were divided into FVH (+ ) group (78 cases) and FVH(-) group (38 cases). All patients underwent magnetic resonance(MR) and MR-PWI scans.Demographic and cerebrovascular risk factors were collected, clinical neurological function of patients was assessed by national institate of health stroke scale(NIHSS) upon admission and discharge, and cognitive function of patients was assessed by mini-mental state examination (MMSE). Short-term clinical outcome was assessed using modified Rankin scale(mRS) at the 90th day after discharge.The degree of middle cerebral artery stenosis, positive or negative FVH, FVH score, hypoperfusion volume and MR-PWI related parameters, including peak time (Tmax), mean transit time (MTT), cerebral blood volume (CBV) and cerebral blood flow (CBF), were evaluated in relation to clinical symptoms.SPSS 22.0 statistical software was used for t test, Chi-square test and Pearson correlation analysis. Results:There were significant differences in hypoperfusion volume, Tmax, MTT and CBF between FVH (+ ) group and FVH(-) group( t=1.989, 3.830, 5.223, 3.911, all P<0.05). In terms of short-term clinical outcome, the improvement rate of neurological function ((8.25±6.39)%, (12.22±6.08)%) and MMSE score(25.48±1.59), (26.31±1.26) in FVH (+ ) group were significantly lower than those in FVH(-) group, and the number of patients with progressive stroke during hospitalization in FVH(+ ) group was more than that of FVH(-) group(22(28.21%), 4(10.53%)) (all P<0.05). Pearson correlation analysis showed that FVH score was positively correlated with hypoperfusion volume ( r=0.786, P<0.01) and MTT ( r=0.692, P<0.01), and negatively correlated with CBF ( r=-0.568, P<0.01), but no significant correlation with the degree of arterial stenosis ( r=0.363, P>0.05). Conclusion:FVH is closely related to the Tmax, MTT and CBF values shown in MR-PWI, and the incidence of stroke in progression and short-term adverse prognosis are more likely in FVH(+ ) group, suggesting that FVH can be used as a convenient imaging indicator to reflect the hypoperfusion status of patients with middle cerebral artery stenosis cerebral infarction, and can provide an objective basis for further individualized treatment.

Objective To develop a self-assessment scale for medication literacy in patients with coronary heart disease comorbidity diabetes and to test its reliability and validity.Methods According to medication literacy theory model,the initial scale was formed through literature review,the qualitative interview and expert inquiry.Cognitive interview was used to optimize the expression of item text.421 patients with coronary heart disease comorbidity diabetes in a tertiary hospital in Zhejiang province from November 2022 to April 2023 were selected to investigate the reliability and validity of the scale by convenience sampling.Results The self-assessment scale of drug literacy for coronary heart disease comorbidity diabetes mellitus included 23 items in 5 dimensions including acquisition,understanding,communication,evaluation and calculation.The total Cronbach's α coefficient of the scale was 0.911;the retest reliability was 0.948;the average content validity index was 0.997;the correlation coefficients between each dimension and total score of the scale and the calibration scale ranged from 0.485 to 0.926.The exploratory factor analysis was employed to extract 5 common factors,and the cumulative variance contribution rate was 73.753％.Confirmatory factor analysis showed that the scale factor structure was stable.Conclusion The scale has good reliability and validity,and it can be used as an effective tool to evaluate the self-rated medication literacy level of patients with coronary heart disease comorbidity diabetes.

Objective:To investigate the influencing factors of the outcome after endovascular therapy (EVT) in patients with large vessel occlusive stroke in anterior circulation.Methods:Patients with large vessel occlusive stroke in anterior circulation received EVT at the Qingpu Branch of Zhongshan Hospital Affiliated with Fudan University and the Fourth Affiliated Hospital of Nanjing Medical University from October 2022 to December 2023 were included retrospectively. The modified Rankin Scale was used for outcome evaluation at 90 days after onset, ≤2 points were defined as good outcome and >2 points were defined as poor outcome. Multivariate logistic regression analysis was used to identify independent influencing factors for poor outcome after EVT. Results:A total of 156 patients with large vessel occlusive stroke in anterior circulation received EVT were enrolled from two stroke centers, including 81 males (51.9%), aged 73.32±9.68 years. Sixty-nine patients (44.2%) had good outcome, 87 (55.8%) had poor outcome. Univariate analysis showed that there were significant differences in the onset-to-door time, door-to-puncture time, puncture-to-reperfusion time, baseline National Institutes of Health Stroke Scale (NIHSS) score, baseline Alberta Stroke Program Early CT Score (ASPECTS), core infarct volume, ischemic penumbra volume, modified Thrombolysis in Cerebral Infarction (mTICI), postprocedural intracranial hemorrhage, and symptomatic intracranial hemorrhage between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that the baseline NIHSS score (odds ratio [ OR] 1.107, 95% confidence interval [ CI] 1.010-1.213; P=0.029), door-to-puncture time ( OR 1.014, 95% CI 1.000-1.028; P=0.048), onset-to-door time ( OR 1.006, 95% CI 1.003-1.009; P<0.001), the core infarct volume ( OR 1.043, 95% CI 1.004-1.084; P=0.029), and mTICI <2b ( OR 10.117, 95% CI 1.896-53.983; P=0.007) were the independent risk factors for poor outcome, while baseline ASPECTS ( OR 0.976, 95% CI 0.961-0.991; P=0.002) and ischemic penumbra volume ( OR 0.359, 95% CI 0.203-0.634; P<0.001) were the independent protective factors for good outcome. Conclusion:For patients with large vessel occlusive stroke in anterior circulation, higher baseline NIHSS score, longer onset-to-door time and door-to-puncture time, larger core infarct volume, and mTICI <2b are associated with the poor outcome after EVT, while higher baseline ASPECTS and larger ischemic penumbra volume are associated with the good outcome.

ObjectiveTo achieve high-dimensional prediction of class imbalanced of adverse drug reaction（ADR） of traditional Chinese medicine（TCM） and to classify and identify risk factors affecting the occurrence of ADR based on the post-marketing safety data of TCM monitored centrally in real world hospitals. MethodThe ensemble clustering resampling combined with regularized Group Lasso regression was used to perform high-dimensional balancing of ADR class-imbalanced data， and then to integrate the balanced datasets to achieve ADR prediction and the risk factor identification by category. ResultA practical example study of the proposed method on a monitoring data of TCM injection performed that the accuracy of the ADR prediction， the prediction sensitivity， the prediction specificity and the area under receiver operating characteristic curve（AUC） were all above 0.8 on the test set. Meanwhile， 40 risk factors affecting the occurrence of ADR were screened out from total 600 high-dimensional variables. And the effect of risk factors on the occurrence of ADR was identified by classification weighting. The important risk factors were classified as follows：past history， medication information， name of combined drugs， disease status， number of combined drugs and personal data. ConclusionIn the real world data of rare ADR with a large amount of clinical variables， this paper realized accurate ADR prediction on high-dimensional and class imbalanced condition， and classified and identified the key risk factors and their clinical significance of categories， so as to provide risk early warning for clinical rational drug use and combined drug use， as well as scientific basis for reevaluation of safety of post-marketing TCM.

Brain metastasis is a common and serious complication of breast cancer, which is commonly associated with poor survival and prognosis. In particular, the treatment of brain metastasis from triple-negative breast cancer (BM-TNBC) has to face the distinct therapeutic challenges from tumor heterogeneity, circulating tumor cells (CTCs), blood-brain barrier (BBB) and blood-tumor barrier (BTB), which is in unmet clinical needs. Herein, combining with the advantages of synthetic and natural targeting moieties, we develop a "Y-shaped" peptide pVAP-decorated platelet-hybrid liposome drug delivery system to address the all-stage targeted drug delivery for the whole progression of BM-TNBC. Inherited from the activated platelet, the hybrid liposomes still retain the native affinity toward CTCs. Further, the peptide-mediated targeting to breast cancer cells and transport across BBB/BTB are demonstrated in vitro and in vivo. The resultant delivery platform significantly improves the drug accumulation both in orthotopic breast tumors and brain metastatic lesions, and eventually exhibits an outperformance in the inhibition of BM-TNBC compared with the free drug. Overall, this work provides a promising prospect for the comprehensive treatment of BM-TNBC, which could be generalized to other cell types or used in imaging platforms in the future.

Objective: To study the expression of long non-coding RNA LINC01152 in glioma and its influence on the malignant biological behavior of glioma cells． Methods: LINC01152 expression in glioma was analyzed by LncSpA V2．0 and GEPIA database．qRT-PCR was applied to detect the expression of LINC01152 mRNA in 10 samples of human normal brain tissues，40 samples of glioma tissues and 5 glioma cell lines．GO and KEGG enrichment analysis of LINC01152 co-expressed genes were performed using the DAVID database to predict the related functions． The AnoLnc2，TargetScan，LinkedOmics and miRDB databases were used to predict the LINC01152 related miRNAs and target genes to construct a ceRNA regulatory network．LINC01152 expression was knocked down in glioma cell lines by small interfering RNA (siRNA) transfection．The CCK-8 test，scratch healing experiments，Transwell，flow cytometry and Western blot experiments were used to measure the influence of LINC01152 on the proliferation，migra- tion，invasion and apoptosis of glioma cells. Results : Database analysis showed that compared with other tumor types，LINC01152 was highly expressed in glioblastoma (GBM) and low grade glioma (LGG) ，and was higher than normal brain tissue．qRT-PCR showed that the expression of LINC01152 mRNA in glioma tissues was significantly higher than that in normal brain tissues (P＜0. 01)．The expression of LINC01152 was correlated with Ki-67 (P < 0. 05) ，but not with clinical parameters such as gender，age，tumor size，P53 protein，glial fibrillary acidic protein (GFAP) ，O-6-methylguanine-DNAmethyltransferase ( MGMT) and WHO grade of glioma patients． Functional enrichment analysis of co-expressed genes indicated that the LINC01152 was mainly involved in biological processes such as cell adhesion and synaptic signaling．LINC01152-miRNA-mRNA regulatory network was constructed according to predicted target genes．After down-regulation of LINC01152 expression，the proliferation，migration and invasion abilities of A172 and U87 cells decreased(P＜0. 01) ，while the apoptosis of glioma cells significantly increased (P＜0. 001) ． Conclusion LINC01152 is highly expressed in glioma，which can promote the malignant biological behavior of glioma cells by enhancing proliferation，migration as well as invasion and inhibition of apoptosis.