Tissue factor pathway inhibitor (TFPI) is the main inhibitor of tissue factor-mediated coagulation.TFPI is expressed by endothelial and smooth muscle celis in the vasculature.Endothelium-derived TFPI has been reported to play a regulatory role in arterial thrombosis.However,the role of endogenous TFPI in vascular smooth muscle cells (VSMCs) in thrombosis and vascular disease development has yet to be elucidated.In this TFPIFlox mice crossbred with Sma-Cre mice were utilized to establish TFPI conditional knockout mice and to examine the effects of VSMC-directed TFPI deletion on development,hemostasis,and thrombosis.The mice with deleted TFPI in VSMCs (TFPIsma) reproduced viable offspring.Plasma TFPI concentration was reduced 7.2％ in the TFPIsma mice compared with TFPFlox littermate controls.Plasma TFPI concentration was also detected in the TFPITie2 (mice deleted TFPI in endothelial cells and cells of hematopoietic origin) mice.Plasma TFPI concentration of the TFPITie2 mice was 80.4％ lower (P ＜ 0.001) than that of the TFPIFlox mice.No difference in hemostatic measures (PT,APTT,and tail bleeding) was observed between TFPIsma and TFPIFlox mice.However,TFPIsma mice had increased ferric chloride-induced arterial thrombosis compared with TFPIFlox littermate controls.Taken together,these data indicated that endogenous TFPI from VSMCs inhibited ferric chloride-induced arterial thrombosis without causing hemostatic effects.

Objective To compare the errors of double-center and single-center setup, and to study the role of both on reducing the rotational setup errors for the patients with esophageal carcinoma depend on rigid registration errors between online kV-cone-beam computed tomography (kV-CBCT) images and plans for CT images. Methods 20 patients with middle esophageal carcinoma received image scanning before treatment every week by using double-center setup and CBCT, and single-center setup images of 20 patients were taken from the X volume image (XVI) system. Then the images of both setup types, registration errors of CT image and rotational setup errors were compared respectively. Every patient received kV-CBCT scanning analysis before treatment every week, and 6 times in total. 240 group of kV-CBCT images from all of the patients were off-line matched with plans for CT images to calculate the errors of X-axis, Y-axis, Z-axis. Then the data of linear errors and rotational setup errors from patients were collected, aiming at putting the error data into the patients treatment program and analyzing the significances. Results The standard registration of double-center setup was as follows: T (X) (0.28 ±0.19) cm, T (Y) (0.27 ±0.19) cm, T (Z) (0.33 ±0.12) cm, R (X) (0.40 ±0.19)° , R (Y) (0.30 ±0.18)° , R (Z) (0.30 ±0.19)° . The standard registration of single-center setup was as follows:T(X) (0.32±0.20) cm, T(Y) (0.29±0.25)cm, T(Z) (0.31±0.16) cm, R(X) (2.2±0.68)°, R(Y) (0.5±0.32)°, R(Z) (2.10±0.60)°. There were statistical differences between linear errors in T(X) and rotational setup errors in R(X), R(Y) or R(Z) (P< 0.05). Conclusion Double-center position can reduce the rotational setup errors, especially in X-axis, Y-axis errors, and may provide more help for the radiation oncology departments without on-board CBCT.

Objective Todeterminetherelationshipbetweenthetumorvolumeoftheperipherallungadenocarcinomawith maximum diameter≤3cmandlymphnodemetastasis(LNM).Methods TheMSCTmanifestationsof235subjectswhowerediagnosedasperipheral lungadenocarcinomawithmaximumdiameter≤3cm wereretrospectivelyanalyzed.Thesepatientsweregroupedaccordingtodifferent parametersincludingsmokinghistory,differentiation,tumorconsistencyandavailabilityoftumornecrosis.Tumorvolumeandratesof LNMamongthesegroupswerecompared.ROCanalysiswasusedtocalculatethecut-offvalueanddiagnosticaccuracy.Results (1) ThetumorvolumeofLNMgroupwaslargerthanthatofnoLNMgroup,cut-offvaluewas5.5cm3,andAUCwas0.76;(2)Therates ofLNMofthewell,moderate-well,moderate,moderate-poorandpoordifferentiationgroupswere0%,8.7%,17.7%,45.6%and46.7%respectively.Theratesofpuregroundglassopacity(p-GGO),mixedandsolidtumorwere0%,8.3%and29.3%respectively.The ratesofthetumorpresentandabsentofnecrosiswere47.8%,22.0%respectively(P＜0.05).Conclusion Usingthevolumeoftumor on MSCTtopredictLNMisanewnon-invasivewayofassessingLNM,withhighsensitivityandspecificity,whichcouldsupplymore imaginginformationforsurgeonstochoosethewayoflymphnodedissection.

Objective To compare the size of target volume,amplitudes of movements in different directions,movement vector,dose to the diseased lung,whole lung volume,and setup error between free breathing fixation (method A) and four-dimensional computed tomography (4DCT)-guided abdominal balloon compression fixation (method B),and to demonstrate that the 4DCT-guided abdominal balloon compression fixation is effective in the treatment of non-small cell lung cancer (NSCLC).Methods A retrospective analysis was performed among 80 patients with NSCLC in our hospital.In those patients,40 received method A and 40 method B.The GTVfree and GTVpress were delineated on the maximum intensity projection (MIP) images of 10 respiratory phases using method A and method B,respectively.The PTVfree and PTVPress were obtained by expansion of the GTVfree and GTVpress,respectively.The paired t test was used to analyze the differences in the PTV,maximum amplitudes of movements in three dimensions,absolute value of the movement vector (|V|),and volume between method A and method B.The treatment planning system was used to compare the V5,V10,V20,and V30 of the diseased lung and the whole lung volume between method A and method B.All patients underwent cone-beam CT (CBCT) scans after positioning.Setup error was obtained by matching the CBCT images with the MIP images in the XVI system based on bone and grayscale values.Results The PTVfree and PTVpress were (283.2± 12.74) and (201.8± 12.99)cm3,respectively (P=0.002).The maximum amplitudes of movements in the right-left,superior-inferior,and anterior-posterior directions as well as thel V | value were (0.22±0.02),(1.85±0.08),(0.43±0.26),and (1.91±0.27) em,respectively,for method A,and (0.05±0.01),(0.41±0.03),(0.16±0.16),and (0.44±0.16) cm,respectively,for method B (P=0.120,0.001,0.070).The V5,V10,V20,and V30 for the diseased lung and total lung volume were (61.26± 4.27) ％,(44.52± 1.70) ％,(28.22± 3.13) ％,(18.26±5.17)％,and (3556±223.12) cm3,respectively,for method A,and (52.74±4.78)％,(38.76±4.92) ％,(23.71 ±4.03) ％,(15.54±3.43) ％,and (3376±311.65) cm3,respectively,for method B (P =0.001,0.003,0.004,0.021,0.004).There was no significant difference in setup error obtained by the XVI system between the two fixation methods (P＞0.05).Conclusions Without increasing setup error,abdominal balloon compression can effectively control the lung movement amplitude,reduce the planning target volume,and reduce the radiation dose to the lung in patients with NSCLC.

Objective:To analyze the cardiac dosimetry of lymph node irradiation in the internal breast region after left-sided breast cancer surgery and to assess its impact on patients' quality of life.Methods:The clinical data of 108 patients who underwent inverse intensity modulated radiotherapy (IMRT) after left-sided breast cancer surgery in Cancer Hospital of Nantong University from May 2019 to May 2021 were collected and retrospectively analyzed, and divided into a study group (with internal breast, 55 cases) and a control group (without internal breast, 53 cases) according to whether the postoperative radiotherapy included lymph node irradiation in the internal breast region. The dosimetric indexes of planned target area (PTV) , cardiac tolerance, serum myocardial injury markers and quality of life before and after radiotherapy were compared between the two groups.Results:In terms of PTV dosimetry, the conformality index (CI) of the study group and the control group were 0.73±0.07 and 0.75±0.08, the homogeneity index (HI) were 0.17±0.03 and 0.17±0.02, the D max were (55.69±1.02) Gy and (55.46±1.13) Gy, the D mean were (50.54±0.23) Gy and (50.48±0.21) Gy respectively, there were no statistically significant differences ( t=1.38, P=0.169; t<0.01, P>0.999; t=1.11, P=0.269; t=1.41, P=0.160) . In terms of cardiac receptivity, the D mean of the two groups were (5.93 ± 0.32) Gy, (5.64 ± 0.30) Gy, V 40 were (0.47 ± 0.10) %, (0.41 ± 0.11) %, and V 30 were (2.48 ± 0.51) %, (2.06 ± 0.49) % respectively, and there were statistically significant differences ( t=4.86, P<0.001; t=2.97, P=0.004; t=4.36, P<0.001) . The levels of serum troponin Ⅰ (cTnⅠ) before radiotherapy in the study group and the control group were (0.09±0.02) ng/ml and (0.09±0.01) ng/ml, creatine kinase isoenzyme MB (CK-MB) were (0.27±0.08) U/L and (0.25±0.08) U/L, myoglobin (MYo) were (3.84±1.02) μg/L and (3.69±0.97) μg/L, and brain natriuretic peptide (BNP) were (172.35±16.24) pg/ml and (169.81±15.93) pg/ml respectively, there were no statistically significant differences ( t<0.01, P>0.999; t=1.30, P=0.197; t=0.78, P=0.436; t=0.82, P=0.414) . One month after radiotherapy, the levels of serum cTnⅠ in the two groups were (0.09±0.03) ng/ml and (0.09±0.02) ng/ml, CK-MB were (0.29±0.09) U/L and (0.28±0.08) U/L, MYo were (4.06±1.08) μg/L and (4.01±1.03) μg/L, and BNP were (175.13±17.09) pg/ml, (172.47±16.28) pg/ml respectively, there were no statistically significant differences ( t<0.01, P>0.999; t=0.61, P=0.544; t=0.25, P=0.806; t=0.83, P=0.410) . The European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) scores before radiotherapy in the study and the control groups were (60.24±5.13) points and (61.19±5.46) points, (74.12±7.20) points and (75.35±7.88) points at 1 month after radiotherapy, (77.53±7.14) points and (78.95±7.08) points at 6 months after radiotherapy, and (75.02±6.93) points and (76.68±6.74) points at 1 year after radiotherapy respectively, there were no statistically significant differences ( t=0.93, P=0.353; t=0.85, P=0.399; t=1.04, P=0.302; t=1.26, P=0.210) . The EORTC QLQ-C30 scores at 1 month, 6 months, and 1 year after radiotherapy were higher than those before radiotherapy in the two groups, and there were statistically significant differences (all P<0.001) . Conclusion:IMRT containing lymph node irradiation in the internal breast region after left breast cancer surgery brings a certain degree of increased cardiac dose, but it is feasible to control it within a certain range and does not affect the patients' cardiac function or quality of life in the short term.

Objective:To establish a cell line stably expressing the transient receptor potential melastatin 2(TRPM2)channel for screening TRPM2 inhibitors based on PiggyBac transposition system.Methods:A plasmid PiggyBac-human TRPM2(pPB-hTRPM2)eukaryotic expression vector was constructed using PiggyBac transposition system.The plasmid and a helper plasmid were co-transfected into HEK293T cells to express TRPM2,which was identified by fluorescence and patch-clamp assays.The high throughput screening performance was assessed with the Z'factor.Calcium imaging and patch clamp techniques were employed to assess the initial activity of eleven compound molecules,confirming the inhibitory effects of the primary molecules on TRPM2.The protective effect of the screened compounds on damaged cells was validated using the oxygen-glucose deprivation/reperfusion(OGD/R)injury model and CCK-8 kit.The level of cellular reactive oxygen species(ROS)was detected by flow cytometry.The neuroprotective effects of the compounds were evaluated using a transient middle cerebral artery occlusion(tMCAO)mouse model.Results:The HEK293T cells transfected with pPB-hTRPM2-EGFP showed high TRPM2 expression.Puromycin-resistant cells,selected through screening,exhibited robust fluorescence.Whole-cell patch results revealed that induced cells displayed classical TRPM2 current characteristics comparable to the control group,showing no significant differences(P＞0.05).With a Z'factor of 0.5416 in calcium imaging,the model demonstrated suitability for high-throughput screening of TRPM2 inhibitors.Calcium imaging and electrophysiological experiments indicated that compound 6 significantly inhibited the TRPM2 channel.Further experiments showed that 1.0 μmol/L of compound 6 enhanced cell viability(P＜0.05)and reduced the level of ROS(P＜0.05)of SH-SY5Y under OGD/R injury.0.3 and 1.0 mg/kg of compound 6 reduced the cerebral infarction volume in tMCAO mice(both P＜0.05).Conclusions:A stable TRPM2 gene expressing cell line has been successfully established using PiggyBac gene editing in this study.TRPM2 channel inhibitors were screened through calcium imaging and patch clamp techniques,and an inhibitor compound 6 was identified.This compound can alleviate cell damage after OGD/R by reducing cellular ROS levels and has a protective effect against cerebral ischemia-reperfusion injury in mice.

This paper summarizes the current situation and necessity of the training of traditional Chinese medicine (TCM) nursing specialists in China, as well as the existing achievements and existing problems, and puts forward the prospects and suggestions for the training of TCM nursing team, so as to strengthen the construction of TCM nursing culture, build a high-quality TCM nursing talent team, and promote the high-quality development of TCM nursing.

Breast cancer remains a leading cause of mortality in women worldwide.Triple-negative breast cancer(TNBC)is a particularly aggressive subtype characterized by rapid progression,poor prognosis,and lack of clear therapeutic targets.In the clinic,delineation of tumor heterogeneity and development of effective drugs continue to pose considerable challenges.Within the scope of our study,high hetero-geneity inherent to breast cancer was uncovered based on the landscape constructed from both tumor and healthy breast tissue samples.Notably,TNBC exhibited significant specificity regarding cell prolif-eration,differentiation,and disease progression.Significant associations between tumor grade,prog-nosis,and TNBC oncogenes were established via pseudotime trajectory analysis.Consequently,we further performed comprehensive characterization of the TNBC microenvironment.A crucial epithelial subcluster,E8,was identified as highly malignant and strongly associated with tumor cell proliferation in TNBC.Additionally,epithelial-mesenchymal transition(EMT)-associated fibroblast and M2 macrophage subclusters exerted an influence on E8 through cellular interactions,contributing to tumor growth.Characteristic genes in these three cluster cells could therefore serve as potential therapeutic targets for TNBC.The collective findings provided valuable insights that assisted in the screening of a series of therapeutic drugs,such as pelitinib.We further confirmed the anti-cancer effect of pelitinib in an orthotopic 4T1 tumor-bearing mouse model.Overall,our study sheds light on the unique characteristics of TNBC at single-cell resolution and the crucial cell types associated with tumor cell proliferation that may serve as potent tools in the development of effective anti-cancer drugs.

Objective To compare the efficacy of endoscopic bougie/balloon dilation(EBD),endoscopic radial incision(ERI),and ERI combined with esophageal stent placement(ESP)for the treatment of benign esophageal stenosis,and evaluate the feasibility and safety of ERI combined with ESP for the treatment of benign esophageal stenosis.Methods 48 Patients with benign esophageal stenosis from January 2019 to January 2023 were recruited,and divided into EBD group(n=24),ERI group(n=17)and ERI+ESP group(n=7).The differences in operating success,restenosis and complications among the three groups were compared.Results The number of previous endoscopic treatment in ERI+ESP group was more than that in EBD group and ERI group,and the differences were statistically significant(P＜0.05).Technical success was achieved in 23 cases and clinical remission in 23 cases in EBD group,technical success in 16 cases and clinical remission in 15 cases in ERI group,technical success in 7 cases and clinical remission in 7 cases in ERI+ESP group.There was no significant difference in technical success rate and clinical remission rate among the three groups(P＞0.05).After 3 months of follow-up,there were 15,9 and 1 cases of esophageal restenosis in the EBD group,ERI group and ERI+ESP group,respectively.There was no significant difference in the rate of esophageal restenosis among the 3 groups(P＞0.05).After 6 months of follow-up,there were 20 cases of esophageal restenosis in the EBD group,13 cases in the ERI group and 1 case in the ERI+ESP group.The rate of esophageal restenosis in the ERI+ESP group was significantly lower than that in the EBD group and the ERI group(P＜0.05).However,there was no statistically significant difference in the esophageal restenosis rate between the EBD group and the ERI group(P＞0.05).The time to the first postoperative restenosis was 74.00(48.75,159.00)days in the EBD group,84.00(54.50,195.00)days in the ERI group,and 250.00(206.00,289.00)days in the ERI+ESP group.The time to the first postoperative restenosis was longer in the ERI+ESP group than that in the EBD and ERI groups.The differences were statistically significant(P＜0.05),but there was no significant difference in restenosis time between EBD group and ERI group(P＞0.05).There were 5,5 and 3 cases of complications in the EBD group,ERI group and ERI+ESP group,respectively,and there was no significant difference in the incidence of complications among the three groups(P＞0.05).Conclusion ERI+ESP is comparable to EBD and ERI in terms of technical success and short-term clinical remission rate for the treatment of benign esophageal stenosis,and is superior to EBD and ERI in terms of long-term restenosis rate and restenosis time,with no influence on the occurrence of complications.

Atrial fibrillation (AF) is the most common arrhythmia in clinic, which can cause hemodynamic changes, heart failure and stroke, and seriously affect human life and health. As a self-promoting disease, the treatment of AF can become more and more difficult with the deterioration of the disease, and the early prediction and intervention of AF is the key to curbing the deterioration of the disease. Based on this, in this study, by controlling the dose of acetylcholine, we changed the AF vulnerability of five mongrel dogs and tried to assess it by analyzing the electrophysiology of atrial epicardium under different states of sinus rhythm. Here, indices from four aspects were proposed to study the atrial activation rule. They are the variability of atrial activation rhythm, the change of the earliest atrial activation, the change of atrial activation delay and the left-right atrial dyssynchrony. By using binary logistic regression analysis, multiple indices above were transformed into the AF inducibility, which were used to classify the signals during sinus rhythm. The sensitivity, specificity and accuracy of classification reached 85.7%, 95.8% and 91.7%, respectively. As the experimental results show, the proposed method has the ability to assess the AF vulnerability of atrium, which is of great clinical significance for the early prediction and intervention of AF.