Objective:To investigate the clinical value of modified acute aortic dissection risk score in the early diagnosis of acute aortic dissection (AAD).Methods:The general, clinical, and imaging data of 162 patients who complained of chest and back pain who received treatment between January 2019 and January 2021 in the Department of Emergency, The Second Hospital of Jiaxing, China were collected for this study. The included patients were divided into control (non-AAD, n = 120) and observation (AAD, n = 42) groups according to whether they were diagnosed with AAD. The indexes with statistical significance between the two groups were analyzed using multivariate logistic regression analysis. A score table was established according to the size of OR value. The modified AAD risk score was predicted using the receiver operating curve. Results:Multivariate logistic regression analysis showed that male sex, family history, sudden severe chest and back pain, bilateral blood pressure asymmetry, hypertension, abnormal ultrasound, and D-dimer level were independent risk factors for the diagnosis of AAD (statistical values = 7.84, 6.96, 7.04, 11.38, 7.12, 8.15, 15.07, 9.11, all P < 0.05). Taking the total score of 5 as the prediction standard, the specificity and sensitivity in the prediction of the occurrence of AAD were 84.94% and 95.43%, respectively. The area under the receiver operating curve regarding the modified AAD risk score was 0.909. Conclusion:The modified AAD risk score can be used to conveniently and quickly predict the occurrence of AAD and has a high predictive value. This study is highly innovative and scientific.

Objective: To construct the third generation chimeric antigen receptor based on a novel humanized anti-HER2 H1-2 scFv, and to investigate the specific cytotoxicity of H1-2 CAR modified T lymphocytes(CAR-T) against HER2⁺ tumor cells. Method: The expression cassette of the third generation CAR gene and anti-HER2 H1-2 scFv were constructed and cloned into lentivirus transfer plasmid, and then the third generation H1-2 CAR was transduced into human T lymphocytes using lentivirus.Enzyme linked immunosorbent assay was used to detect the expression of cytokines IL2, and LDH release assay was used to detect the cytotoxic effect of the H1-2 CAR-T.Finally, NOD/SCID mice and HER2⁺ breast cancer cell line SKBR3 were used to detect the anti-tumor effect of H1-2 CAR-T in vivo. Results: The third generation H1-2 CAR was successfully constructed.H1-2 CAR-T secreted high dose of IL2 after confrontation with HER2⁺ breast cancer cells.In vitro, the cytolytic rate of H1-2 CAR-T on high expression HER2⁺ tumor cells was significantly higher than that in low expression HER2 or non-expression HER2 tumor cells. At the efficacy to target ratio of 20, the cytolytic rate of H1-2 CAR-T against breast cancer cell SK-BR-3 could reach (90.1±2.8)%, while the cytolytic rate of H1-2 CAR-T against HER2^- breast cancer cell MDA-MB-231 was only (13.5±4.7)%. In the mouse xenograft tumor model, H1-2 CAR-T cells inhibited breast cancer growth in vivo.At the end of the experiments, the average tumor weight in the H1-2 CAR-T cell treatment group was (0.7±0.1) g, the non-transfected T cell therapeutic group was (1.2±0.2) g, and the PBS group was (1.2±0.2) g. There was significant difference between the H1-2 CAR-T therapeutic group and the non-transfected T cell therapeutic group (P<0.05). However, there was no significant difference between the non-transfected T cell therapeutic group and the PBS treatment group (P>0.05). Conclusion The HER2-sepcific H1-2 CAR-T cells specifically kill HER2 positive cells, and further studies on CAR-T cells for the treatment of HER2⁺ cancers are useful.

Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.

Objective To investigate the dynamic changes of symptom clusters among nasopharyngeal carcinoma patients receiving radiotherapy and provide evidence for intervention clinically.Methods The data were collected at three time points that were before radiotherapy,during the radiotherapy and after radiotherapy, by surveying 273 first visit of nasopharyngeal carcinoma patients from May 201 4 to January 201 5 with a self-designed general information questionnaire and the Chinese version of the M.D.Anderson symptom inventory and observing the changes of symptom clusters in our hospital.One-way analysis of variance and the exploratory factor analysis were applied to observe the incidence of symptoms and the changes of symptom clusters during radiotherapy.Results The statistics revealed that the incidence and intensity of all the symptoms at 3 time points were significantly increased(P <0.05),except the symptom of amnesia.There were 3 symptom clusters among 3 time points,named as gastrointestinal symptom cluster,sickness symptom cluster and somatic symptom cluster,but the composition and the variance were different at different time point.Gastrointestinal symptom cluster was made of nausea and vomiting,adding poor appetite only before radiotherapy.The gastrointestinal symptom cluster was most prominent before radiotherapy.Sickness symptom cluster consisted of drowsiness, numbness symptom before radiotherapy, while it consisted of shortness of breath, numbness, difficulty remembering symptoms in other time.Somatic symptom cluster mainly consisted of pain and fatigue,dry mouth symptom joining it during and after radiotherapy.Conclusions With the treatment of radiotherapy,the incidence and intensity of symptoms are significantly enhanced,and the constitution of symptom clusters is different at different stage.Therefore,it is important for clinical nurses to strengthen dynamic assessment of all symptoms,monitor the changes and composition of symptom clusters,thus establish corresponding management measures to improve patients′quality of life.

BackgroundPatients after percutaneous coronary intervention （PCI） are prone to experience depression， which has been shown to significantly affect patients' quality of recovery in postoperative period. Resourcefulness plays an important role between stress and depression， yet there is insufficient research evidence on the predictive effect of intellectual resourcefulness levels on depressive symptoms among patients undergoing PCI. ObjectiveTo investigate the cross-lagged effect between level of resourcefulness and depressive symptoms among patients after PCI， so as to provide references for alleviating depressive symptoms in patients after PCI. MethodsA total of 363 patients who had undergone PCI in Tangshan Gongren Hospital from September to December 2019 were selected using random sampling technique. All participants were subjected to complete Chinese Resourcefulness Scale （CRS） and Chinese version of Cardiac Depression Scale （C-CDS） at both baseline and two years after PCI. A structural equation model was constructed to determine the cross-lagged effect between the level of resourcefulness and depressive symptoms in patients. ResultsMale patients scored higher on CRS （t=-19.871， P<0.01） and lower on C-CDS （t=25.557， P<0.01） after two years of PCI compared with female patients after PCI. Correlation analysis indicated that the baseline CRS score was positively correlated with two years after PCI CRS score （r=0.550， P<0.01）， C-CDS score of baseline was positively correlated with two years after PCI C-CDS score （r=0.524， P<0.01）， baseline CRS score was negatively correlated with C-CDS scores at both baseline （r=-0.717， P<0.01） and two years after PCI （r=-0.472， P<0.01）， and two years after PCI， CRS score was negatively correlation with C-CDS score （r=-0.618， P<0.01）. The cross-lagged analysis revealed that baseline CRS score significantly predicted CRS score of two years after PCI （β=0.382， P<0.01） and C-CDS score of two years after PCI （β=-0.200， P<0.01）. Baseline C-CDS score significantly predicted C-CDS score of two years after PCI （β=0.381， P<0.01） and CRS score of two years after PCI （β=-0.235， P<0.01）. There was a reciprocal relationship between baseline CRS score and baseline C-CDS score （β=-0.717， P<0.01）. ConclusionThe established cross-lagged model yields the presence of a reciprocal prediction of level of resourcefulness and depressive symptoms measured in patients at two time points. The higher the baseline level of resourcefulness， the lighter the depressive symptoms experienced by patients two years after PCI surgery.The more severe the baseline depressive symptoms， the lower the patients' level of resourcefulness two years after PCI surgery.［Funded by Project of Hebei Provincial Department of Science and Technology （number， 182777154）］

Objective:To evaluate the feasibility of 8E5 cells and CD19-CAR-Jurkat cells used as reference cells in the detection of lentiviral vector integration sites with different methods.Methods:Single clones of 8E5 cells and CD19-CAR-Jurkat cells were selected using limiting dilution method. Digital PCR was established to detect the copy number of HIV-1 in 8E5 cells and the copy number of CAR in CD19-CAR-Jurkat cells. High-throughput sequencing techniques (whole-genome resequencing, modified genome sequencing and probe hybridization capture) were used to detect integration sites in 8E5 cells and CD19-CAR-Jurkat cells, and optical genome mapping (OGM) technology was used for further confirmation.Results:Three clones of 8E5-D8 cells and six clones of CD19-CAR-Jurkat 2-6 cells were selected using the limiting dilution method. 8E5-D8 and CD19-CAR-Jurkat 2-6 were chosen as candidate cells based on their gene copy numbers detected by digital PCR and flow cytometry. These cells were then expanded and cryopreserved. Digital PCR showed that 8E5-D8 cells contained approximately 1 copy per cell, while CD19-CAR-Jurkat 2-6 cells contained approximately 13 copies per cell. High-throughput sequencing revealed one integration site in 8E5 cells and 13 integration sites in CD19-CAR-Jurkat cells, which matched the copy number detection results. All these integration sites were further confirmed at the submicroscopic level of chromosomes using OGM.Conclusions:Based on the insertion copy numbers and integration sites, 8E5-D8 cells and CD19-CAR-Jurkat 2-6 cells could be used as reference cells in further development of methods for detecting integration sites in CAR-T cell lentiviral vectors.

Objective: To facilitate using the CRISPR/Cas9 gene editing system in human liver and gallbladder cancer cells, we established Cas9 stably expressed human liver and gallbladder cancer cell lines, and validated the gene editing activity of Cas9. Methods: Human liver cancer cell lines (Huh7, PLC/PRF/5, HepG2, Hep3b, SK-HEP-1 and Li-7), human cholangiocarcinoma cells (RBE) and human gallbladder cancer cells (GBC-SD) were infected with 3 Cas9-expressing lentivirus vectors (pLv-EF1α-Cas9-Flag-Neo, pLv-EF1α-Cas9-Flag-Puro, Cas9m1.1), respectively, and Cas9 stably expressed colonies were screened and selected. We extracted the genomic DNA and protein, validated the stable expression of Cas9 by using genomic polymerase chain reaction (PCR) and western blot. Three of cell lines were further infected with Lv-EF1α-mCherry. Then mCherry positive cells were sorted by flow cytometry and infected with designed guide RNA (gRNA) vectors which targeted mCherry gene. Subsequently the gene editing activity of Cas9 was detected by genomic PCR, fluorescence microscopic observation and flow cytometry analysis. Results: One hundred Cas9-expressing human liver and gallbladder cancer cell lines were selected. Among them, 35 cell lines expressed Cas9-Neo, 25 expressed Cas9-puro, and 40 expressed mutant Cas9 (mCas9). We also established 3 cell lines with stable expression of mCherry (Huh7-mCas9-M, PLC/PRF/5-Cas9-M and SK-HEP-1-Cas9-M). The results of genomic PCR and sequencing showed that by lentiviral infection with 2 types of designed gRNA, the long fragment deletion of mCherry gene was found in these 3 cell lines. Moreover, mCherry^-EGFP⁺ cells infected with 2 types of gRNA were observed by fluorescence microscope. The results of flow cytometry showed that mCherry^-EGFP⁺ cells accounted from 0.3% to 93.6%. Conclusion We successfully establish 100 human liver and gallbladder cancer cell lines with stable expression of Cas9 protein and validate their activities of gene editing.