Sini Decoction (SNT) is a traditional formula recognized for its efficacy in warming the spleen and stomach and dispersing cold. However, elucidating the mechanism of action of SNT remains challenging due to its complex multiple components. This study utilized a synergistic approach combining two-dimensional fluorescence difference in gel electrophoresis (2D-DIGE)-based drug affinity responsive target stability (DARTS) with label-free quantitative proteomics techniques to identify the direct and indirect protein targets of SNT in myocardial infarction. The analysis identified 590 proteins, with 30 proteins showing significant upregulation and 51 proteins showing downregulation when comparing the SNT group with the model group. Through the integration of 2D-DIGE DARTS with proteomics data and pharmacological assessments, the findings indicate that protein disulfide-isomerase A3 (PDIA3) may serve as a potential protein target through which SNT provides protective effects on myocardial cells during myocardial infarction.
Myocardial Infarction/genetics* ; Proteomics/methods* ; Drugs, Chinese Herbal/chemistry* ; Animals ; Protein Disulfide-Isomerases/genetics* ; Male ; Two-Dimensional Difference Gel Electrophoresis/methods* ; Humans ; Rats ; Rats, Sprague-Dawley ; Electrophoresis, Gel, Two-Dimensional

Objective To investigate the prognostic value of peripheral blood neutrophil-lymphocyte ratio(NLR)and thioredoxin reductase(TrxR)in patients with ovarian cancer receiving immunotherapy.Methods A total of 109 patients with advanced ovarian cancer treated in the Tumor Hospital Affiliated to Nantong University from January 2021 to December 2021 were selected as the research objects.The levels of NLR and TrxR in peripheral blood before immunotherapy were detected,and the evaluation value of NLR and TrxR on short-term efficacy,progression-free survival(PFS)and overall survival(OS)in ovarian cancer pa-tients receiving immunotherapy was explored.Results The optimal cut-off values of TrxR and NLR were 4.97 U/mL and 2.49%,respectively.According to the optimal cut-off value of TrxR and NLR,the patients were divided into the high level of TrxR group(69 cases,≥4.97 U/mL)and the low level of TrxR group(40 cases,<4.97 U/mL),the high level of NLR group(72 cases,≥2.49%)and the low level of NLR group(37 cases,<2.49%).The objective response rate(ORR)of the high level of NLR group was lower than that of the low level NLR group(P<0.05),and the disease progression rate(DPR)was higher than that of the low NLR group(P<0.05).The high level of TrxR group had a significantly lower ORR and a significantly higher DPR than the low level of TrxR group(P<0.05).The median PFS and OS of the high level of NLR group were 15.0 months and 16.0 months,respectively.The median PFS and OS of the low level of NLR group were 19.0 months and 21.0 months,respectively.The median PFS and OS of the high level of TrxR group were 15.0 months and 17.0 months,respectively.The median PFS was 18.0 months and the median OS was 21.0 months in the low level of TrxR group.NLR and TrxR were the influencing factors of PFS and OS in pa-tients with ovarian cancer immunotherapy(P<0.05).Conclusion The levels of NLR and TrxR in peripheral blood can be used as important prognostic indicators for advanced ovarian cancer patients receiving immuno-therapy.The lower the levels of NLR and TrxR,the better the prognosis of ovarian cancer patients.

Objective To investigate the expression of circular RNA ubiquitin-associated protein 2(circ-UBAP2)and RAS-related C3 botulinum toxin substrate 1(Rac1)in cervical cancer tissues and their relation-ship with radiotherapy efficacy and prognosis.Methods A total of 96 patients with cervical cancer admitted to the hospital from January 2019 to May 2021 were selected as the research subjects.Detect the expression of circ-UBAP2 and Rac1 in the cancer tissues and adjacent tissues of the patients.The clinical data of the patients were recorded.According to the radiotherapy efficacy,they were divided into patients with effective radiother-apy(n=62)and patients with ineffective radiotherapy(n=34),and the relationship between the expressions of circ-UBAP2 and Rac1 and the radiotherapy efficacy was analyzed.The survival conditions of the patients were followed up for 3 years and they were divided into the survival group(n=81)and the death group(n=15),and the clinical data of the two groups were compared.The relationship between the expressions of circ-UBAP2 and Rac1 and the survival prognosis of patients was analyzed by Kaplan-Meier.The risk factors influ-encing the prognosis of patients were analyzed by Cox regression.Results The circ-UBAP2 expression and the positive rates of Rac1 expression in cancer tissues were both higher than those in adjacent tissues,and the difference was statistically significant(P＜0.05).The effective rate of radiotherapy in patients with high ex-pression of circ-UBAP2 and Rac1 in cancer tissues was lower than that in patients with low expression of circ-UBAP2 and Rac1,and the difference was statistically significant(P＜0.05).The 3-year overall survival rate and average survival duration of patients with high expression of circ-UBAP2 and Rac1 in cancer tissues were lower than those of patients with low expression of circ-UBAP2 and Rac1,and the difference was statistically significant(P＜0.05).There were statistically significant differences in clinical stage,tumor differentiation degree,expressions of circ-UBAP2 and Rac1,and the proportion of lymph node metastasis between the two groups(P＜0.05).The results of Cox regression analysis showed that clinical stage Ⅱ B-Ⅲ,poorly differen-tiated tumor degree,high expression of circ-UBAP2 and Rac1,and lymph node metastasis were risk factors af-fecting the prognosis of patients(P＜0.05).Conclusion circ-UBAP2 and Rac1 are abnormally highly ex-pressed in cervical cancer tissues and are related to the radiotherapy efficacy and prognosis.

Objective To evaluate the imaging features of posterior circulation ischemic stroke caused by vertebral artery dissection(VAD)by high-resolution magnetic resonance imaging(HR-MRI).Methods A total of 56 patients with highly suspected VAD were selected.According to HR-MRI characteristics,these patients were divided into ischemic stroke group(n=23)and control group(n=33).The correlation between imaging features and risk factors in the two groups was analyzed.Results The proportions of intramural hematoma,tumor-like dilatation,and degree of vessel wall enhancement in the ischemic stroke group were significantly higher than those in the control group,and the difference was statistically significant(P<0.05).However,there was no statistical significance in the double-chamber sign and intimal valve sign(P=0.075).Correlation analysis showed that the effective lumen index was significantly negatively correlated with the incidence of posterior circulation ischemic stroke in patients with VAD(r=-0.721,P<0.05),and the area under the curve(AUC)of the effective lumen index reached 0.935.Conclusion The effective features of HR-MRI in the diagnosis of posterior circulation ischemic stroke caused by VAD include intramural hematoma,tumor-like dilatation,degree of vessel wall enhancement,and reduction of effective lumen index,which are helpful for the management and prediction of patients with VAD.

Objective:To investigate the effect of bone mesenchymal stem cells derived exosomes (BMSC-Exo) on improving hippocampal microangiogenesis, energy metabolism, and behaviors in depression mouse models.Methods:(1) Mouse bone marrow mesenchymal stem cells were isolated and cultured to extract BMSC-Exo; BMSC-Exo morphology was observed by transmission electron microscopy, BMSC-Exo particle diameter ranges were determined by Zetaview analyzer, and expressions of CD9 and CD63 in BMSC-Exo were detected by Western blotting. (2) Depression models were established in 2 mice by chronic unforeseeable mild stress (CUMS); 24 h after stereotaxic injection of phosphate buffer solution (PBS) or DiR labeled BMSC-Exo, BMSC-Exo uptake was detected by in vivo imaging system. (3) Thirty-six mice were randomly divided into control group, model group and BMSC-Exo group ( n=12); CUMS was used to establish depression models in the latter 2 groups; brain stereotaxic injection of 1 μL BMSC-Exo was given to mice in the BMSC-Exo group after modeling, and same amount of PBS was given to the control group; behaviors were observed by forced swimming test (FST), tail suspension test (TST) and open field test (OFT); hippocampal microvascular length and number were detected by alkaline phosphatase staining; energy metabolism in the hippocampus was detected by micro positron emission tomography/computed tomography (mPET/CT); glucose transporter 1 (GLUT1) expression in the hippocampus was detected by Western blotting. Results:(1) BMSC-Exo had a typical disk-like vesicle-like structure with particle size of (100.5±1.4) nm; Western blotting confirmed that CD9 and CD63 expressed in BMSC-Exo. (2) In vivo imaging showed no fluorescence in the brain and liver after PBS injection, but obvious local fluorescence after BMSC-Exo injection. (3) Compared with the control group, the model group and BMSC-Exo group had significantly longer rest time in FST and TST and shorter movement distance and time in the central region of OFT ( P<0.05); compared with the model group, BMSC-Exo group had significantly shorter rest time in FST and TST and longer movement distance and time in the central region of OFT ( P<0.05). Compared with the control group, the model group and BMSC-Exo group had significantly decreased standard uptake value (SUV) of regions of interest, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05); compared with the model group, the BMSC-Exo group had significantly higher SUV, microvascular length and number, and GLUT1 expression in the hippocampus ( P<0.05). Positive correlations were noted between hippocampal microvascular length and SUV and between microvascular number and SUV in the 3 groups ( r=0.540, P<0.001; r=0.600, P<0.001). Conclusion:BMSC-Exo could promote microangiogenesis energy metabolism in the hippocampus to improve depression-like behaviors in depression mouse models.

Objective:To explore the association between changes in brain structural network during the early stage of stroke recovery and the onset of post-stroke depression (PSD).Methods:A total of 87 acute ischemic stroke patients scheduled for discharge, who were admitted to the Yixing Hospital Affiliated to Jiangsu University from March 2020 to May 2021, were prospectively collected. During the same period, 34 healthy control subjects matched with the stroke patients were also collected. All participants underwent systematic magnetic resonance imaging scans and scale assessments, and were followed up longitudinally for 2 years. Based on the occurrence of depression during follow-up, the stroke patients were divided into PSD group and post-stroke non-depression (PSND) group. Graph theoretical analysis was used to analyze the topological characteristics of brain structural network. Analysis of variance was used to explore the differences in brain structural network attributes among groups. Logistic regression model was used to analyze the predictive power of differential brain network attributes for PSD. Linear regression analysis was conducted to investigate the relationship between the synergism of non-rich-club regions and changes in rich-club connectivity.Results:The rich-club connectivity and synergism of the non-rich-club regions were significantly lower in the PSD group than in the PSND group (rich-club connectivity, P<0.01; synergism of feeder/local, P<0.001). The regression model demonstrated that the synergism of non-rich-club regions had a good predictive power for the occurrence of PSD ( OR=1.195, 95%CI 1.073-1.471, P<0.001). Furthermore, linear regression analysis revealed a significant correlation between the synergism of non-rich-club regions and Δrich-club connectivity ( r=-0.691, P<0.001). Conclusion:The good synergism of non-rich-club regions during the early stage of stroke recovery promotes the repair of rich-club connectivity and inhibits the onset of PSD.

Objective:To investigate the features of the brain structural network in patients with postpartum depression (PPD).Methods:This cross-sectional study included PPD patients who visited the mental health counseling clinic after delivery at the Jiangsu University Affiliated Yixing Hospital from June 2013 to September 2022 (PPD group). Matched non-PPD postpartum women based on age, years of education, and body mass index who came for postpartum follow-up (non-PPD postpartum group), and non-pregnant women who visited the hospital or underwent physical examinations during the same period (non-pregnant group) were also included. Demographic data and diffusion tensor imaging (DTI) data were collected for all three groups. The brain was partitioned into 90 regions using an anatomical template to construct the brain structural network. Network-based statistics (NBS) were applied to further screen and construct subnetworks. The efficacy of the subnetworks in identifying PPD was evaluated through multivariable logistics regression models and receiver operating characteristic curves. A comparison of the connectivity strength of white matter tracts and topological attributes of brain structural network parameters was conducted using independent samples t-tests, and the results were corrected using the false discovery rate (FDR) method. Results:(1) A total of 116 subjects were included, with 40 in the non-pregnant group, 40 in the non-PPD postpartum group, and 36 in the PPD group. PPD group had higher Edinburgh Postnatal Depression Scale (EPDS) scores than the non-pregnant and non-PPD postpartum groups [(18.0±4.1) scores vs. (2.5±1.2) and (6.1±2.1) scores, F=340.40; t=24.65,10.60 and 16.16 in pairwise comparison; all P<0.001]. (2) Compared to the non-pregnant group, there was a decrease in the connectivity strength of nine white matter tracts within the brain structural network of the postpartum group (including left dorsolateral superior frontal gyrus-left anterior cingulate and paracingulate gyrus, left dorsolateral superior frontal gyrus-right amygdala, left dorsolateral superior frontal gyrus-left insula, left insula-left lentiform nucleus, left insula-left hippocampus, left hippocampus-right amygdala, left hippocampus-left precuneus, left anterior cingulate and paracingulate gyrus-right amygdala, and right amygdala-right hippocampus) (all P<0.05, FDR corrected). No increased connection strengths were observed. There were no significant differences in the connection strengths of these nine tracts between the non-PPD and PPD groups. (3) A characteristic subnetwork for the maternal group was successfully constructed based on the nine tracts, which exhibited typical small-world properties (σ>1). Compared to the non-PPD maternal group, the characteristic path length in the PPD group was increased [(3.904±0.328) vs. (4.130±0.433), t=－2.58], and global efficiency was decreased [(0.361±0.036) vs. (0.331±0.053), t=2.91] (both P<0.05). Local property comparisons showed that the node efficiency values for the left dorsolateral superior frontal gyrus, left insula, left anterior cingulate and paracingulate gyrus, left hippocampus, right hippocampus, right amygdala, left precuneus and left putamen in the PPD group were significantly reduced [(0.273±0.023) vs. (0.267±0.030), t=0.98; (0.299±0.035) vs. (0.276±0.041), t=2.64; (0.265±0.019) vs. (0.258±0.025), t=1.38; (0.318±0.028) vs. (0.305±0.031), t=1.92; (0.312±0.027) vs. (0.302±0.031), t=1.50; (0.322±0.030) vs. (0.298±0.026), t=3.71; (0.356±0.040) vs. (0.338±0.056), t=1.62; (0.346±0.028) vs. (0.331±0.036), t=1.74; all P<0.05]. However, only the differences in node efficiency values for the left insula and right amygdala remained significant after FDR correction (corrected P=0.041 and 0.003). (4) Global efficiency, as well as node efficiency for the left insula and right amygdala, demonstrated good value for identifying PPD [areas under the curve (AUC) and their 95% CI were 0.827 (0.732-0.922), 0.741 (0.628-0.854), and 0.761 (0.653-0.867), respectively], with even better performance when combined [0.897 (0.828-0.969)]. (5) In the PPD group, global efficiency ( r=－0.43, P=0.008), node efficiency for the left insula ( r=－0.39, P=0.019), and node efficiency for the right amygdala ( r=－0.42, P=0.011) were all negatively correlated with EPDS scores. Conclusion:Aberrations in global efficiency, node efficiency for the left insula, and node efficiency for the right amygdala may serve as characteristic neuroimaging biomarkers for PPD.

Objective:To investigate the effects and significance of acute and chronic trauma on brain degree centrality (DC) in patients with post-traumatic stress disorder (PTSD) who lost their only child at resting state.Methods:Retrospectively, the study enrolled a total of 51 parents with PTSD, including 35 PTSD parents whose children was lost in emergencies (acute bereaved PTSD group) and 16 PTSD parents whose children was lost of chronic causes such as diseases (chronic bereaved PTSD group). Fifty local adults were also included as healthy controls (HC group). The clinical administered PTSD scale(CAPS) was used to evaluate the severity of the subjects' clinical symptoms.Resting-state functional magnetic resonance imaging(fMRI) data of all subjects were collected and DC values were calculated.SPSS 22.0 software was used for statistical analysis.Covariance analysis was performed among three groups, while post hoc was performed between any two groups.What's more, correlation analyses were utilized between abnormal brain regions and the scores of CAPS.Results:Significant group effects were found in multiple regions, including the right inferior temporal gyrus (MNI: x, y, z=66, -27, -21), right temporal pole (MNI: x, y, z=54, 15, -9), right orbital inferior frontal gyrus (MNI: x, y, z=42, 21, -15), bilateral medial superior frontal gyri (MNI: right x, y, z=6, 63, 12; left x, y, z=-3, 60, 18), left inferior parietal angular gyrus (MNI: x, y, z=-45, -36, 51) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51). Compared with HC group, the DC of two patient groups increased in the right inferior temporal gyrus (MNI: acute x, y, z=63, -27, -21; chronic x, y, z=63, -21, -27); the DC of acute bereaved PTSD group decreased in the right temporal pole (MNI: x, y, z=45, 21, -15) and the right orbital inferior frontal gyrus (MNI: x, y, z=48, 24, -12), while the DC of chronic bereaved PTSD group decreased in the left inferior parietal angular gyrus (MNI: x, y, z=-45, -36, 51) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51). Compared with chronic bereaved PTSD group, the DC of acute bereaved PTSD group increased in the left inferior parietal angular gyrus (MNI: x, y, z=-33, -39, 42) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51), while decreased in the right temporal pole (MNI: x, y, z=51, 12, -9), right orbital inferior frontal gyrus (MNI: x, y, z=42, 21, -15) and bilateral medial superior frontal gyri (MNI: left x, y, z=0, 57, 15; right x, y, z=3, 57, 15). In chronic bereaved PTSD group, the DC of the left postcentral gyrus was negatively correlated with C1 (avoid trauma-related thoughts, feelings) score in CAPS ( r=-0.606, P=0.028). In acute bereaved PTSD group, the DC of the left medial superior frontal gyrus was negatively correlated with D4 (high vigilance) score ( r=-0.416, P=0.020). Conclusion:There exist functional abnormalities of multiple brain regions in acute and chronic bereaved parents with PTSD.The high arousal symptoms of the former may be related with the abnormalities of prefrontal-amygdala neural circuit, while the latter show higher avoidance which may be associated with the dysfunction of somatosensory brain regions such as postcentral gyrus.

Cell membrane affinity chromatography has been widely applied in membrane protein (MP)-targeted drug screening and interaction analysis. However, in current methods, the MP sources are derived from cell lines or recombinant protein expression, which are time-consuming for cell culture or purification, and also difficult to ensure the purity and consistent orientation of MPs in the chromatographic stationary phase. In this study, a novel in situ synthesis membrane protein affinity chromatography (iSMAC) method was developed utilizing cell-free protein expression (CFE) and covalent immobilized affinity chromatography, which achieved efficient in situ synthesis and unidirectional insertion of MPs into liposomes in the stationary phase. The advantages of iSMAC are: 1) There is no need to culture cells or prepare recombinant proteins; 2) Specific and purified MPs with stable and controllable content can be obtained within 2 h; 3) MPs maintain the transmembrane structure and a consistent orientation in the chromatographic stationary phase; 4) The flexible and personalized construction of cDNAs makes it possible to analyze drug binding sites. iSMAC was successfully applied to screen PDGFRβ inhibitors from Salvia miltiorrhiza and Schisandra chinensis. Micro columns prepared by in-situ synthesis maintain satisfactory analysis activity within 72 h. Two new PDGFRβ inhibitors, salvianolic acid B and gomisin D, were screened out with K _D values of 13.44 and 7.39 μmol/L, respectively. In vitro experiments confirmed that the two compounds decreased α-SMA and collagen Ӏ mRNA levels raised by TGF-β in HSC-T6 cells through regulating the phosphorylation of p38, AKT and ERK. In vivo, Sal B could also attenuate CCl₄-induced liver fibrosis by downregulating PDGFRβ downstream related protein levels. The iSMAC method can be applied to other general MPs, and provides a practical approach for the rapid preparation of MP-immobilized or other biological solid-phase materials.

P-glycoprotein (P-gp) highly expressed in cancer cells can lead to multidrug resistance (MDR) and the combination of anti-cancer drugs with P-gp inhibitor has been a promising strategy to reverse MDR in cancer treatment. In this study, we established a label-free and detergent-free system combining surface plasmon resonance (SPR) biosensor with styrene maleic acid (SMA) polymer membrane proteins (MPs) stabilization technology to screen potential P-gp inhibitors. First, P-gp was extracted from MCF-7/ADR cells using SMA polymer to form SMA liposomes (SMALPs). Following that, SMALPs were immobilized on an SPR biosensor chip to establish a P-gp inhibitor screening system, and the affinity between P-gp and small molecule ligand was determined. The methodological investigation proved that the screening system had good specificity and stability. Nine P-gp ligands were screened out from 50 natural products, and their affinity constants with P-gp were also determined. The in vitro cell verification experiments demonstrated that tetrandrine, fangchinoline, praeruptorin B, neobaicalein, and icariin could significantly increase the sensitivity of MCF-7/ADR cells to Adriamycin (Adr). Moreover, tetrandrine, praeruptorin B, and neobaicalein could reverse MDR in MCF-7/ADR cells by inhibiting the function of P-gp. This is the first time that SMALPs-based stabilization strategy was applied to SPR analysis system. SMA polymer can retain P-gp in the environment of natural lipid bilayer and thus maintain the correct conformation and physiological functions of P-gp. The developed system can quickly and accurately screen small molecule ligands of complex MPs and obtain affinity between complex MPs and small molecule ligands without protein purification.