Tumor-infiltrating lymphocytes (TILs) are a heterogeneous subset of lymphocytes, mainly T cells, present in tumor parenchyma and stroma. After being digested and isolated from tumor tissue and then cultured in vitro for activation and multiplication, it can be infused back into the patient's body to kill tumor cells. TILs have the advantages of high diversity of TCR, excellent ability to infiltrate into tumor sites, and low toxicity and are considered promising for the treatment of malignant solid tumors. At present, TIL therapy has been tested as a second-line treatment in a variety of solid tumors and has achieved preliminary results. Although there is still no clinical cohort report on the application of TILs in biliary tract cancer (BTC), recent clinical reports on multiple cancers have provided information on the efficacy of TIL therapy in a small number of BTC patients, which preliminarily confirmed the safety and efficacy of TIL therapy. However, since BTC is generally considered an immunologically repulsive tumor in which most effector T cells are sequestered at the tumor edge, the antitumor effect of TILs in BTC remains difficult to predict. Combination therapy with different anti-tumor methods and the development of new techniques to modify cells to enhance the anti-tumor ability of TILs are possible directions for breakthrough in the future.

Objective:To compare the early effect of water sac blocking and Heartstring for proximal anastomosis of the calcific ascending aorta.Methods:The data of 400 consecutive patients undergoing elective off-pump coronary artery bypass grafting(OPCABG) in Beijing Anzhen Hospital from January 2022 to June 2022 were retrospectively analyzed. 46 patients with calcific ascending aorta including 40 males and 6 females, with the age ranged from 53 to 73 years and an average of(65.2±5.1) years, who were revealed by preoperative chest CT scan and intraoperative palpation. According to the method of proximal anastomosis, the patients were divided into 2 groups: water sac blocking group(n=19) and Heartstring group(n=27). The effect of preventing postoperative stroke was compared by counting the incidence of postoperative stroke. The efficacy of the 2 methods was compared by detecting the flow and pulsatility fraction of the saphenous vein trunk during surgery, observing the dynamic changes of the electrocardiogram and cTnI level within 48h after the surgery, and reviewing the coronary CTA 3 months after discharge.Results:There was no perioperative death, and all the patients were discharged 4-13 days postoperatively. No adverse events such as stroke and malignant ventricular arrhythmia occurred during perioperative period. 1 patient in each group developed low cardiac output syndrome postoperatively, and both improved after IABP placement. 1 patient in Heartstring group developed acute inferior myocardial infarction, which was improved after IABP placement. Coronary CTA 3 months after operation showed that there was no proximal anastomotic stenosis in both groups.Conclusion:There is no significant difference between the 2 proximal anastomosis methods in preventing stroke after OPCABG in patients with ascending aortic calcification. Compared with Heartstring, water sac blocking does not increase the risk of proximal anastomotic stenosis. In addition, water sac blocking does not require expensive consumables, which is especially suitable for patients with limited funds and can be generalized.

Objective:To investigate the effects of NOD-like receptor protein 3(NLRP3) inflammasome activation on the proliferation, migration and extracellular matrix desposition of activated pancreatic stellate cells(PSCs).Methods:The rat PSCs were isolated, cultured and identified, and were divided into control group or LPS group based on the pretreatment with LPS (10 μg/ml for 24 hours) or without. The expression of NLRP3 inflammasome associated molecules in PSCs culture medium was detected by ELISA. The PSCs with NLRP3 inhibition were constructed by shRNA carrying lentivirus infection and were divided into LPS+ negative control group and LPS+ lentivirus group based on whether the cells were treated with LPS and infected by lentivirus or not. The alteration in cell proliferation and migration were detected by CCK-8 kit and transwell chamber method. The expression of extracellular matrix α-SMA and collagen in PSCs was detected by immunofluorescence staining and the expression of TGF-β mRNA was analyzed by RT-qPCR.Results:The cytoplasm of PSCs which were cultured for 24 hours was rich in bright annular lipid droplets, and the cells expressed desmin. After 7 days of culture, the cell became larger in size, the lipid droplets basically disappeared, and the cells were activated and expressed α-SMA. The expression of caspase-1, IL-1β and IL-18 in the supernatant of PSCs culture medium in LPS group were significantly higher than those in control group (1.55±0.04 vs 0.65±0.03), (2.02±0.04 vs 1.05±0.05) and (1.70±0.05 vs 0.97±0.03), respectively. After inhibiting by lentivirus infection, the expression of NLRP3 in the lentivirus group (0.25±0.04) was significantly lower than that in negative control group (0.68±0.05). In control group, LPS group, LPS+ negative control group and LPS+ lentivirus group, the A490 values was 0.61±0.02, 1.15±0.06, 0.96±0.05, and 0.56±0.01, respectively; the migrating PSCs number was (64.12±4.58), (121.67±8.02), (111.67±4.67) and (69.67±8.08)/HF, respectively; the relative expression of α-SMA was 0.78±0.05, 4.12±0.04, 3.81±0.06 and 0.88±0.05, respectively; the relative expression of collagen was 0.65±0.03, 3.43±0.02, 2.67±0.02 and 0.48±0.03, respectively; and the expression of TGF-β mRNA was 0.22±0.03, 0.89±0.01, 0.86±0.03 and 0.43±0.02, respectively. The A490 value, the migrating cells number, the expression of α-SMA, collagen and the expression of TGF-β mRNA in LPS group and LPS+ negative control group was significantly higher than those in control group and LPS+ lentivirus group, and all the differences were statistically significant (all P value <0.05). Conclusions:NLRP3 inflammasome activation may accelerate the extracellular matrix deposition and pancreatic fibrogenesis by promoting PSCs proliferation and migration ability via regulating the biological functions.

Objective:To investigate the application value of CT and MRI imageomics based on machine learning method in the diagnosis of pancreatic cancer.Methods:The clinical data of 62 patients with surgically resected and pathologically confirmed pancreatic cancer, who underwent enhanced CT scan, MRI plain or enhanced scan in Shanghai General Hospital between January 2014 and December 2021 were collected. According to the chronological order of surgery, 49 patients from January 2014 to December 2020 were enrolled in the training set and 13 patients from January 2021 to December 2021 were enrolled in the validation set. 3D-slicer 4.8.1 software was used to draw the region of interest in each layer of CT and MRI images for cancerous and paracancerous tissue segment. Image features were extracted by Python and the optimal feature set from the training set data was obtained by using Lasso regression model. The machine learning decision tree model was constructed. The receiver operating characteristic curve(ROC) curve was drawn, and the area under the curve (AUC) was calculated to evaluate the value of these three kinds of imageomics models in the diagnosis of pancreatic cancer.Results:The 1 767 CT features and 1 674 MRI features were obtained from enhanced CT scan, MRI plain scan and enhanced MRI scan, respectively. For the differential diagnosis model of cancerous tissue and paracancerous tissue, the enhanced CT scan data model obtained the optimal feature set involving 6 features, the MRI plain scan model obtained the optimal feature set involving 16 features, and the enhanced MRI scan model obtained the optimal feature set involving 15 features. The diagnostic model based on enhanced CT scan had an AUC of 0.98 in the training set and 1 in the verification group. The AUC of the MRI plain scan and enhanced MRI scan models in both the training set and the validation set was 1. The specificity and sensitivity of machine learning decision tree model based on the three kinds of imageomics models in the diagnosis of cancerous tissue and paracancerous tissue were 100%. For the differential diagnosis model of splenic artery wrapping, the enhanced CT scan model didn′t obtain the optimal features and had no diagnostic efficacy. The MRI plain scan model and enhanced MRI scan model obtained the optimal feature set involving 5 and 4 features, respectively. The AUC of the MRI plain scan model in the training set and the validation set were 0.862 and 0.750, respectively, with diagnostic sensitivity of 93.8% and 50.0%, and specificity of 78.6% and 100%, respectively. The AUC of the enhanced MRI scan model in the training set and the validation set were 0.950 and 0.861, respectively, with diagnostic sensitivity of 90.0% and 93.6%, and specificity of 100% and 78.6%, respectively.Conclusions:Based on the radiomics of CT enhanced, MRI plain scan and enhanced MRI scan, the machine learning diagnostic model has an accuracy of more than 90% in differentiating pancreatic cancer from paracancerous tissue. For the differentiation of splenic artery wrapping in pancreatic cancer, the diagnostic model based on enhanced MRI scan haS the best diagnostic efficiency.

Objective:To investigate the value of large-scale brain network research based on independent component analysis (ICA) in discovering the changes of intrinsic functional connections within and between resting-state networks (RSNs) in first-episode untreated adult patients with major depressive disorder (MDD).Methods:From January 2019 to June 2021, twenty-three patients with MDD (MDD group) and 30 healthy volunteers (HC group) matched with gender, age and years of education were selected. All participants underwent resting-state brain function imaging (rs-fMRI), and the MDD group completed the 17-item Hamilton depression scale(HAMD-17). The independent component analysis (ICA) method was used to analyze rs-fMRI data, and meaningful RSNs were obtained. SPM12 and Gift softwares were used to compare the strength of intrinsic functional connection within and between the RSNs of the MDD group and HC group, and the Pearson correlation analysis was conducted by IBM SPSS statistics 25.0 to evaluate the correlation between the functional connection strength and HAMD-17 scores in MDD group.Results:Compared with the HC group, intrinsic functional connection strength of medial prefrontal cortex (mPFC) (MNI: x, y, z=-6, 54, 25)in MDD group was significantly enhanced, while the intrinsic functional connection strength of the left angular gyrus (AG) (MNI: x, y, z=-48, -66, 21), the left precuneus (PCu) (MNI: x, y, z=-6, -63, 33), the left dorsolateral prefrontal cortex (dlPFC) (MNI: x, y, z=-36, 12, 51)and the right anterior insula (AI) (MNI: x, y, z=36, 21, 0)were significantly weakened. Compared with the HC group, functional connection strength between posterior default mode net work(pDMN) and anterior default mode network(aDMN) in MDD group was significantly weakened ( t=-2.206, P=0.032), and function connection strength between pDMN and left frontal parietal network(lFPN) was significantly strengthened ( t=2.318, P=0.025). In MDD group, intrinsic functional connection strength of mPFC and the functional connection strength of pDMN-lFPN were positively correlated with the HAMD-17 score ( r=0.524, P=0.010; r=0.441, P=0.035). Conclusion:Large-scale brain network study based on the ICA can find abnormal functional connections within and between RSNs in first-episode untreated adult patients with MDD, and provide objective imaging markers for the clinical diagnosis and treatment of MDD.

Chest trauma is one of the most common injuries. Venous thromboembolism (VTE) as a common complication of chest trauma seriously affects the quality of patients′ life and even leads to death. Although there are some consensus and guidelines on the prevention and treatment of VTE at home and abroad, the current literatures lack specificity considering the diagnosis, treatment and prevention of VTE in patients with chest trauma have their own characteristics, especially for those with blunt trauma. Accordingly, China Chest Injury Research Society and editorial board of Chinese Journal of Traumatology organized relevant domestic experts to jointly formulate the Chinese expert consensus on the diagnosis, treatment and prevention of chest trauma venous thromboembolism associated with chest trauma (2022 version). This consensus provides expert recommendations of different levels as academic guidance in terms of the characteristics, clinical manifestations, risk assessment, diagnosis, treatment, and prevention of chest trauma-related VTE, so as to offer a reference for clinical application.

ObjectiveTo determine the quality of life among HIV-infected individuals and risk factors in Jing’an District of Shanghai. MethodsThis is a cross-sectional study. Cluster sampling strategy was used to select HIV-infected individuals who were registered with the national HIV/AIDS case reporting system in eight community health service centers in Jing’an District of Shanghai. The short form 36 （SF-36） scale， self-rating anxiety scale （SAS）， social support scale （SSS） and specifically designed questionnaires were used to collect information. Logistic regression was used to determine the factors associated with quality of life. ResultsIn total， 365 HIV-infected individuals were included in the study. The total average score of quality of life was 79.5±10.4. The scores in general health， vitality and emotional well-being were lower than the norm， whereas the scores in other dimensions were higher than the norm. Multivariable logistic regression indicated that age of 45‒ years （vs age of 18‒ years： OR=0.23， 95%CI： 0.08‒0.68） was significantly associated with the quality of life. Furthermore， smoking （OR=2.18， 95%CI： 1.22‒3.90）， lower CD4⁺T cell counts （<350 cell·μL^-1， OR=2.89， 95%CI： 1.54‒5.40）， comorbidity （≥2 kinds， OR=3.40， 95%CI： 1.15‒10.04） and anxiety （OR=1.09， 95%CI： 1.05‒1.14） were risk factors associated with declining quality of life. ConclusionIt warrants more attention to those HIV-infected individuals with poor psychological status， smoking， low CD4⁺T cell counts and comorbidities. Targeted health care and social support could effectively improve the quality of life among HIV-infected individuals.

Objective: To investigate the prevalence of scoliosis and identify the influencing factors among children and adolescents in Jiaxing City, Zhejiang Province, so as to provide insights into the management of scoliosis among children and adolescents. Methods: The fourth, fifth and sixth grade primary school students and the first, second and third grade junior high school students were selected using the stratified cluster sampling method in Jiaxing City in 2019. Students' demographic features, diet habits, physical activity and learning environments and habits were collected using questionnaire surveys. Scoliosis was screened using whole-spine X-ray scans in an upright position, and the factors affecting scoliosis were identified using multivariable logistic regression analysis. Results: Valid surveys were completed among 2 568 students, including 1 352 boys ( 52.65% ) and 1 216 girls ( 47.35% ), and there were 1 335 primary school students ( 51.99% ) and 1 233 junior high school students ( 48.01% ). A total of 93 students were detected with scoliosis, with a prevalence rate of 3.62%. Multivariable logistic regression analysis identified age ( OR=0.826, 95%CI: 0.710-0.960 ), gender ( OR=2.079, 95%CI: 1.343-3.221 ) and vigorous physical activity in the past 7 days ( OR=2.514, 95%CI: 1.248-5.063 ) as factors affecting scoliosis among primary and junior high school students. Conclusions The prevalence of scoliosis is high among primary and junior high school students in Jiaxing City. Age, gender and physical activity may affect the risk of scoliosis. Scoliosis screening is recommended to be included in routine healthy

Objective:To investigate the early effect of different antithrombotic therapy in patients with coronary endarterectomy(CE) combined with off-pump coronary artery bypass grafting(OPCABG).Methods:Between January and December 2021, 154 consecutive patients including 120 males and 34 females with the age ranged from 39 to 78 years and an average of(62.6±7.2) years who underwent CE+ OPCABG were evaluated retrospectively. According to the postoperative anticoagulant therapy, patients were divided into two groups: Aspirin+ low molecular weight heparin group(n=81, LMWH group) and Aspirin+ ticagrelor group(n=73, ticagrelor group). The data of both preoperative and postoperative hemoglobin level and blood transfusion after the surgery were collected. The dynamic changes of electrocardiogram and cTnI level were observed within 48 h after the surgery.Results:There was no perioperative death, and all the patients were discharged 5-13 days postoperatively. After the initiation of anticoagulant therapy, the lowest hemoglobin value in the LMWH group and ticagrelor group was(88.3±14.6)g/L vs.(89.5±11.6)g/L( P>0.05), blood transfusion was performed in 8 vs. 5 patients with hemoglobin below 70g/L( P>0.05), peak cTnI within 48 h of surgery was 850.55(410.63, 1 662.63)pg/ml vs. 1 184.60(667.50, 3 169.63)pg/ml( P<0.05), the number of patients with perioperative myocardial infraction within 48h after the surgery confirmed by electrocardiogram was 2(2.5%) vs.2(2.5%), P>0.05. Conclusion:There was no significant difference between the two anticoagulant treatments in preventing perioperative myocardial infarction after CE+ OPCABG surgery. LMWH did not increase the risk of postoperative bleeding compared with ticagrelor. In addition, aspirin+ LMWH reduced the levels of peak TnI within 48 h of surgery, which may be associated with better long-term postoperative outcomes, but further research is needed to confirm this.

The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)‍-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.
CRISPR-Associated Protein 9/metabolism* ; CRISPR-Cas Systems ; DNA ; Gene Editing/methods* ; Humans ; Streptococcus pyogenes/metabolism*