This study intends to establish a colorectal cancer(CRC)organoid model,expand and isolate CRC-reactive tumor-infiltrating lymphocytes(TILs),screen tumor-specific T cell receptors(TCRs)and perform functional verification,in order to provide a technological platform and research foundation for the clinical transformation of individualized adoptive T-cell immunotherapy for colorectal cancer.An organoid model derived from colon cancer patient tissues was constructed using in vitro 3D culture techniques,which then subjected to HE staining and immunohistochemistry for detecting morphological characteristics and representative molecular expression.Subsequently,CRC organoids were co-cultured with TILs for sorting reactive TILs using flow cytometry,and the characteristics of reactive TCR clones was analyzed through single T cell receptor gene cloning technology.Furthermore,the function of TCRs was verified through cytotoxicity experiments.Morphological analysis and representative molecules(CK20 and CDX2)expression indicated that there is high similarity between colorectal cancer organoids and patient tumors.In the in vitro expanded and cultured TILs,colorectal cancer-reactive T cells with upregulated CD137 expression and increased IFN-γ secretion were screened out successfully,among which TCR2-T cells demonstrated superior tumor reactivity and in vitro tumor killing function.In conclusion,a platform for screening and function validation of reactive TCRs based on CRC-Org has been established,providing a technological platform for the translational application of individualized T-cell therapy for colorectal cancer.

Objective To investigate the clinical characteristics and risk factors of plastic bronchitis(PB)occurrence in children patients with refractory Mycoplasma pneumoniae pneumonia(RMPP).Methods A retrospective analysis was performed on the clinical data of 399 children patients with RMPP treated by fiberoptic bronchoscopy hospitalized in this hospital from January 2017 to December 2019.The pa-tients were divided into the PB group(n=142)and non-PB group(n=257)according to whether or not find-ing PB under fiberoptic bronchoscopy.The differences in clinical characteristics,laboratory detection indicators and imageological manifestations were compared between the two groups.The risk factors of PB occurrence in children RMPP were analyzed.Results Compared with the non-PB group,the incidence rates of hypoxemia and extrapul-monary complications,and the highest body temperature in the PB group were higher,number of fever days and hospitalization days was longer,the proportions of hormone and intravenous injection of immunoglobulin were higher,the levels of NEUT,CRP,IL-6,AST,ALT LDH CK and D-Dimer and incidence rates of pulmo-nary atelectasis and pleural effusion were higher,the levels of PLT and lymphocytes were lower,and the differences were statistically significant(P＜0.05).The receiver operating characteristic(ROC)curve analy-sis results showed that the highest body temperature,NEUT,PCT,IL-6,AST and LDH could serve as the predictive indicators for PB occurrence in RMPP(P＜0.05).The multivariate logistic regression analysis re-sults showed that the highest body temperature＞39.8 ℃,NEUT＞72.9％,IL-6＞26.65 pg/mL,AST＞49.5 U/L and pulmonary atelectasis were the risk factors of PB occurrence in RMPP.Conclusion Should pay at-tention to the risk factors of PB occurrence in children patients with RMPP and take necessary preventive measures to improve their prognosis.

Objective:To investigate the early clinical characteristics of elderly patients with severe burns and the risk factors on prognosis.Methods:This study was a retrospective case series study. Clinical data of 124 elderly patients with severe burns who met the inclusion criteria and were admitted to the 12 hospitals from January 2015 to December 2020 were collected, including 4 patients from the Fourth People's Hospital of Dalian, 5 patients from Fujian Medical University Union Hospital, 22 patients from Guangzhou Red Cross Hospital of Jinan University, 5 patients from Heilongjiang Provincial Hospital, 27 patients from the First Affiliated Hospital of Naval Medical University, 9 patients from the First Affiliated Hospital of Nanchang University, 10 patients from Affiliated Hospital of Nantong University, 9 patients from Tongren Hospital of Wuhan University & Wuhan Third Hospital, 12 patients from the 924 th Hospital of PLA, 6 patients from Zhangjiagang First People's Hospital, 4 patients from Taizhou Hospital of Zhejiang Province, and 11 patients from Zhengzhou First People's Hospital. The patients' overall clinical characteristics, such as gender, age, body mass index, total burn area, full-thickness burn area, inhalation injury, causative factors, whether combined with underlying medical diseases, and admission time after injury were recorded. According to the survival outcome within 28 days after injury, the patients were divided into survival group (89 cases) and death group (35 cases). The following data of patients were compared between the two groups, including the basic data and injuries (the same as the overall clinical characteristics ahead); the coagulation indexes within the first 24 hours of injury such as prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time, D-dimer, fibrinogen degradation product (FDP), international normalized ratio (INR), and fibrinogen; the blood routine indexes within the first 24 hours of injury such as white blood cell count, platelet count, neutrophil-to-lymphocyte ratio, monocyte count, red blood cell count, hemoglobin, and hematocrit; the organ function indexes within the first 24 hours of injury such as direct bilirubin, total bilirubin, urea, serum creatinine, aspartate aminotransferase, alanine aminotransferase, total protein, albumin, globulin, blood glucose, triglyceride, total cholesterol, alkaline phosphatase, creatine kinase, electrolyte indexes (potassium, sodium, chlorine, calcium, magnesium, and phosphorus in blood), uric acid, myoglobin, and brain natriuretic peptide; the infection and blood gas indexes within the first 24 hours of injury such as procalcitonin, C-reactive protein, pH value, oxygenation index, base excess, and lactate; treatment such as whether conducted with mechanical ventilation, whether conducted with continuous renal replacement therapy, whether conducted with anticoagulation therapy, whether applied with vasoactive drugs, and fluid resuscitation. The analysis was conducted to screen the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns. Results:Among 124 patients, there were 82 males and 42 females, aged 60-97 years, with body mass index of 23.44 (21.09, 25.95) kg/m 2, total burn area of 54.00% (42.00%, 75.00%) total body surface area (TBSA), and full-thickness burn area of 25.00% (10.00%, 40.00%) TBSA. The patients were mainly combined with moderate to severe inhalation injury and caused by flame burns. There were 43 cases with underlying medical diseases. The majority of patients were admitted to the hospital within 8 hours after injury. There were statistically significant differences between patients in the 2 groups in terms of age, total burn area, full-thickness burn area, and inhalation injury, and PT, APTT, D-dimer, FDP, INR, white blood cell count, platelet count, urea, serum creatinine, blood glucose, blood sodium, uric acid, myoglobin, and urine volume within the first 24 hours of injury (with Z values of 2.37, 5.49, 5.26, 5.97, 2.18, 1.95, 2.68, 2.68, 2.51, 2.82, 2.14, 3.40, 5.31, 3.41, 2.35, 3.81, 2.16, and -3.82, respectively, P<0.05); there were statistically significant differences between two groups of patients in whether conducted with mechanical ventilation and whether applied with vasoactive drugs (with χ2 values of 9.44 and 28.50, respectively, P<0.05). Age, total burn area, full-thickness burn area, serum creatinine within the first 24 hours of injury, and APTT within the first 24 hours of injury were the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns (with odds ratios of 1.17, 1.10, 1.10, 1.09, and 1.27, 95% confidence intervals of 1.03-1.40, 1.04-1.21, 1.05-1.19, 1.05-1.17, and 1.07-1.69, respectively, P<0.05). Conclusions:The elderly patients with severe burns had the injuries mainly from flame burns, often accompanied by moderate to severe inhalation injury and enhanced inflammatory response, elevated blood glucose levels, activated fibrinolysis, and impaired organ function in the early stage, which are associated with their prognosis. Age, total burn area, full-thickness burn area, and serum creatinine and APTT within the first 24 hours of injury are the independent risk factors for death within 28 days after injury in this population.

This study was performed to explore the impact of glutamine(Gln)on the anti-tumor immune response of CD8+ T cells and its mechanism.TCGA database was used to analysis the relationship between tumor Gln metabolism and the quantity and functionality of infiltrating CD8+ T cells.CRISPR/Cas9 was employed to knock down GLS expression in mouse MC38 cells,and a mouse tumor model was established.Flow cytometry was conducted to assess tumor proliferation,apoptosis,and the quantity and functionality of tumor-infiltrating immune cells.Lymphocytes isolated from health individuals were treated with Gln-deficient media,complete media or media supplemented with GSH,RSL3 in vitro.Then the apoptosis,the expression levels of GPX4,Lipid-ROS,and effector function protein of CD8+ T cells were detected by flow cytometry.Furthermore,RNA-seq was performed to analyze the differential gene expression on the Gln-depleted CD8+ T cells.Data showed that tumor Gln metabolism was inversely associated with the quantity and functionality of tumor-infiltrating CD8+ T cells.Low expression of GLS in MC38 cells could inhibit C57BL/6 tumor growth,decrease Ki-67 expression,promote casepase-3 expression,increase the amount of tumor-infiltrating immune cells,suppress PD-1,TIM-3,and LAG-3 expression,and enhance CD137,CD107a,IFN-γ and TNF-α expression in tumor-infiltrating CD8+ T cells.RNA-seq results indicated an upregulation of ferroptosis genes TFRC,HMOX1,CYBB and SLC7A11 in CD8+ T cells following glutamine deficiency.Gln deficiency led to lower CD137,CD107a,IFN-γ,GSH,GPX4 expression,increased Lipid-ROS level,and caused cell death in CD8+ T cells.Supplementation of GSH upregulated GPX4 expression,downregulated Lipid-ROS level,and increased IFN-γ secretion in CD8+ T cells.In conclusion,Gln deficiency inhibits the effector function of CD8+ T cells by inducing ferroptosis,and promotes tumor growth.

Objective: To explore the correlation between different nutritional status and depressive symptoms among adolescents, so as to provide a theoretical basis for targeted improvement of nutritional status and depressive symptoms in adolescents. Methods: A total of 8 102 adolescents aged 12-17 years in Shanghai, Urumqi, Changsha and Kunming were selected by random cluster sampling. Depressive symptoms were assessed by the Center for Epidemiologic Studies Depression Scale(CES-D). Height and weight were measured. Kruskal Wallis H test, Chi square test and ordinal Logistic regression were used to analyze the relationship between nutritional status and depressive symptoms in adolescents. Results: The overall detection rate of depressive symptoms in adolescents was 25.6%, with girls (30.1%) higher than that of boys (21.1%) ( χ 2=87.60, P <0.01). There were statistically significant differences in the scores of depressive symptoms among girls with different nutritional status ( Z =8.34, P <0.05). The detectable rate of depressive symptoms increased with BMI among girls( χ 2 trend =6.04, P <0.05). After controlling for confounding factors, ordinal Logistic regression showed that the obese girls had higher risks in depressive symptoms detection compared with normal weight girls ( OR=1.45, 95%CI=1.10-1.91, P <0.05). Conclusion The nutritional status of girls is better than boys, but the prevalence of depressive symptoms is higher than boys. The risk of depressive symptoms in girls increases with BMI.

Objective: To explore the correlation between non suicidal self injury(NSSI) behaviors and depressive symptoms in adolescents and to provide a theoretical basis for improvement of adolescent physical and mental health. Methods: A total of 8 102 adolescents aged 12-17 years were selected by random cluster sampling method in Shanghai, Urumqi, Changsha and Kunming, using the Center for Epidemiologic Studies Depression Scale (CES-D) and the Adolescent Non Suicidal Self Injurious Behavior Assessment Questionnaire. Depressive symptoms and the occurrence of NSSI behaviors were assessed. The relationship between NSSI behaviors and depressive symptoms was analyzed using Chi square test, one way analysis and Logistic regression analysis. Results: The overall detection rate of NSSI in adolescents, 24.6%, with a significantly lower detection rate in boys (23.1%) than in girls (26.2%) ( χ 2=10.97, P <0.05), and 24.7%, 24.6% in the middle school and high school segments, respectively, but the difference was not statistically significant ( χ 2=0.01, P >0.05). The overall detection rate of adolescent depressive symptoms was 49.7 % in the group with NSSI and 17.7% in the group without NSSI, with statistically significant differences ( χ 2=808.80, P < 0.01 ). The detection rates of adolescent depressive symptoms in the group with NSSI (male:44.0%, female:54.7%, middle school:49.6%, high school:49.8%) and the group without NSSI (male:14.2%, female:21.4%, middle school:14.3%, high school: 21.0 %) were statistically significant by sex and age groups ( χ 2=385.58, 412.44, 520.60, 313.78, P ＜0.01). Logistic regression models, after adjusting for gender and age, the risk for depressive symptoms was 2.65 times (95% CI =2.27-3.09) and 7.28 times (95% CI = 6.34- 8.37) more frequently in adolescents with episodic self injurious behavior and frequent self injurious behavior, respectively, compared to those without self injurious behavior. The association between NSSI status and depressive symptoms did not show sex differences ( P >0.05). Conclusion Adolescent NSSI behavior is positively associated with depressive symptoms, the attention should be paid to adolescent physical and mental health to reduce the occurrence of NSSI.

Objective: To analyze the correlation between physical activity and depressive symptoms in adolescents, so as to provide a reference for the early prevention of depression among adolescents. Methods: Using a random cluster sampling method, 8 102 adolescents from Shanghai, Urumqi, Changsha and Kunming were investigated via the Physical Activity Assessment Questionnaire of Chinese Children and Adolescents aged 7 to 18 and the Center for Epidemiologic Studies Depression Scale (CES-D) for physical activity and depressive symptoms. Multivariate Logistic regression analysis was used to analyze the correlation between physical activity and depressive symptoms in adolescents. Results: The proportion of adolescents who met moderate to vigorous physical activity (MVPA) standard was 37.0%, and the detection rate of depressive symptoms was 25.6%. The detection rate of depressive symptomsin compliant with MVPA standard group was significantly lower than that in non compliant group among different age and gender( χ 2=7.62-34.54, P <0.05). Adolescents with severe depressive symptoms had high light intensity physical activity(LPA) duration and low vigorous intensity physical activity(VPA) duration, while adolescents with mild depressive symptoms or asymptomatic adolescents had higher MVPA duration. There were differences in LPA and VPA among adolescents with different levels of depressive symptoms in the overall sample, and the differences were still significant among girls, middle school and statistically significant differences in LPA, moderate intensity physical activity(MPA), MVPA were also found among the high school subgroup. However, differences only in LPA were observed among boys with different levels of depression( H=46.48, 10.31, 25.02, 29.54, 30.25, 19.80, 22.20, 9.54, 8.62, 18.06, P <0.05). Multivariate Logistic regression model showed that gender and duration of MVPA were significantly related to risk of depressive symptoms( OR=1.56, 1.42, P <0.05). Conclusion Physical activity among adolescents is significantly related to depressive symptoms. MVPA may be an effective measure to reduce the degree of depressive symptoms.To improve the physical health and prevent depression among adolescents, it is important to choose appropriate physical activity types and effectively increase the duration of MVPA.

Benign gallbladder diseases are common diseases in general surgery, including gallstones, polypoid lesions, cholecystitis, etc. In China, the treatment of benign gallbladder diseases is still not standardized, which mainly shows in following aspects: (1)the tendency of cholecystectomy to all gallbladder diseases may result in the incidence of abdominal pain, distension, diarrhea and bile duct injury after surgery; (2)the attitude of "no-surgery" in which the standard cholecystectomy fails to implement in time for the benign gallbladder diseases with potential high-risk factors for gallbladder cancer, as a result, patients develop occurrence of gallbladder cancer. Especially, gallbladder sparing surgery is an unscientific surgery for benign gallbladder diseases, the gallbladder may become a high-risk factor for cancer after the operation. Implementation of standard cholecystectomy in time for benign gallbladder diseases with potential high risk gallbladder cancer can not only significantly reduce the incidence of gallbladder cancer, but also significantly improve the early diagnosis and the prognosis. Based on current literatures, the authors analyze the treatment status of benign gallbladder diseases, and investigate the disputes and problems in surgical indications, operation time and the selection of treatments of benign gallbladder diseases.

Objective: To investigate the short-term efficacy and adverse events of chemotherapy combined with androgen-deprivation therapy in high-volume metastatic hormone sensitive prostate cancer. Methods: From March 2015 to August 2017, 55 patients with high-volume metastatic hormone sensitive prostate cancer were enrolled at Department of Urology, Fudan University Shanghai Cancer Center receiving chemotherapy combined with androgen-deprivation therapy. The age was 65(8) years (M(Q_R)) (range: 46 to 79 years). Patients were enrolled in the study for continuous androgen-deprivation therapy (medical or surgical castration), combined with docetaxel 75 mg/m² intravenous injection on the first day, repeated every 21 days (6 cycles). Endpoints included overall survival, progression-free survival of prostate cancer, prostate specific antigen (PSA) response rate, and adverse events. Results: The follow-up time was 21.2(11.7) months. The PSA value before chemotherapy was 144.9(415.3) μg/L. The days in patients undergoing androgen deprivation therapy before chemotherapy was 14(23) days. Four patients (7.3%) presented 0 in Eastern Cooperative Oncology Group scoring system and 51 patients(92.7%) presented 1. Thirty-nine patients (70.9%) completed more than 6 cycles of combined chemotherapy, 17 patients (30.9%) showed PSA<0.2 μg/L at 6 months after treatment, and 14 patients (25.5%) showed PSA<0.2 μg/L at 12 months after treatment. Twenty-eight patients (50.9%) had grade 3 to 4 neutropenia and 1 patient (1.8%) developed infectious neutropenia and died. Nausea and vomit occurred in 16 patients (29.1%). Twelve patients (21.8%) underwent dose adjustment due to adverse events in blood system. Conclusions The short-term effect was confirmed in high-volume metastatic hormone sensitive prostate cancer using chemotherapy combined androgen-deprivation therapy, and the long-term effect remains to be seen. Myelosuppression during chemotherapy requires close attention, and taking timely examination is recommended.

Objective To evaluate the therapeutic effects of the accordion technique used in external fixation for long bone hypertrophic nonunion.Methods From August 2013 to December 2015,13 patients with hypertrophic nonunion were treated by the accordion technique (distraction,compression and re-distraction) used together with an Ilizarov ring external fixator or a monolateral external fixator.They were 10 males and 3 females with an average age of 44.6 years (from 18 to 63 years).The nonunion occurred in the middle femur in 5 patients,in the femoral supracondyle in one,in the tibial diaphysis in 4,and in the distal tibia in 3.The procedure was performed with a preassembled external fixator without open surgery or bone graft.The pathological sites were gradually distracted (0.25 mm/day) from the seventh day postoperatively.In case of deformity,correction was conducted to restore the alignment.At the same rate of distraction,compression was started after the equal length was achieved and next followed by distraction.The frequency of cycle depended upon the callus observed in the X-rays.Results The mean follow-up for the 13 patients was 19 months (from 10 to 38 months) after removal of the external fixator.All the cases of hypertrophic nonunion obtained bony union after a mean duration of 7.5 months (from 5 to 13 months).No nonunion or deformity recurred during the follow-up.The complications such as pin-site infection (in 4 cases) and mild Achilles's tendon contracture (in 2 cases) were prevented or treated by careful nursing and rehabilitation.Conclusion The accordion technique together with an external fixator is a reliable and effective treatment for hypertrophic nonunion.