Objective:To construct pancreatic cancer cell lines expressing luciferase and mesothelin (MSLN), and evaluate the feasibility of using them as target cells in analyzing the cytotoxicity activity of immune cells.Methods:Lentiviral vectors expressing luciferase and MSLN genes were constructed, and pancreatic cancer cell lines were infected after lentivirus packaging. Single-cell clones were obtained by limited dilution following antibiotic screening, and the stable expression of the target genes were verified. These cells were used as target cells to detect the cytotoxicity of immune cells by real-time cell analysis (RTCA) and luciferase activity. Besides, these luciferase-expressing cells were transplanted into B-NDG mice to establish the animal models of pancreatic cancer, and in vivo optical imaging technology was used to detect the expression of luciferase and monitor the tumor growth in mice. The cytotoxicity of chimeric antigen receptor T (CAR-T) cells was verified in these animal models. Results:Three pancreatic cancer cell lines, panc-1-luc, panc-1-luc-MSLN and capan-2-luc, that could stably express luciferase and MSLN genes were successfully constructed. The expression of the reporter gene in these cells were high, and positively correlated with the number of cells. There were 95.6% of panc-1-luc-MSLN cells expressing MSLN. MSLN-CAR-T cells had specific killing effect on MSLN-positive panc-1-luc-MSLN cells and capan-2-luc cells, with the minimum killing rates of (70.00±18.19)% and (57.00±5.29)%, respectively. But they had no cytotoxicity to MSLN-negative panc-1-luc cells. RTCA results showed that MSLN-CAR-T cells were able to lyse all three pancreatic cancer cell lines, and the minimum killing rates were (56.33±7.64)%, (93.00±2.65)% and (26.33±28.15)%, respectively. The killing of target cells by NK-92MI cells was not depended on MSLN expression. The cytotoxicity in the mice models of pancreatic cancer was consistent with the results in vitro. The in vivo and in vitro test results suggested that the expression of luciferase by target cells could reflect the cytotoxicity of immune cells. Conclusions:This study establishes three pancreatic cancer cell lines stably expressing luciferase, which can be used to evaluate the cytotoxicity of immunotherapy products targeting tumor cells in vitro and in vivo.

ObjectiveTo explore the typical behavior characteristics of children with autism in symbolic play and the value of Symbolic Play Test (SPT) in the early identification of autism. MethodsFrom November, 2021 to September, 2022, a total of 260 children with language problems were collected from Department of Children's Health Care of Binzhou Medical University Hospital. A total of 193 children with autism were as observation group and 67 normal children were as control group. All children played symbolic games. The typical behavioral characteristics of children with autism in SPT were explored, and a reliability and validity analysis based on the results of SPT was conducted. They were assessed with the adaptive and personal social scores of Gesell Development Scale, and the correlation of the scores of Gesell Development Scale and the score of SPT was analyzed. ResultsThe Cronbach's α coefficient of SPT of children with autism was 0.835 to 0.935, and the total score of SPT, the scores of surrogate object, fictional attribute and fictional object were positively correlated with each other (r > 0.607, P < 0.001). The SPT scores decreased in the observation group (t > 9.615, P < 0.001), and SPT score positively correlated with adaptability and personal-social development quotient (r > 0.609, P < 0.001). ConclusionTypical behavior of children with autism can be reflected in symbolic play, and SPT can provide clues for early identification of autism.

Animals ; Genetics, Population ; history ; History, 20th Century ; History, 21st Century ; Human Genetics ; history ; Humans

Objective To explore the hearing screening, and the change and outcome of hearing impairment of high risk infants. Meth-ods From March, 2015 to March, 2016, 336 high risk infants were screened with otoacoustic emissions (OAE), auditory brainstem response (ABR) and brainstem auditory evoked-potential (BAEP) 0-1, 3, 6, 12 months after born, respectively. Results Among the 336 high risk in-fants, 29 failed the examinations within the 1st month, 37 cases failed in the 3rd month, 27 cases recovered in the 9th month, and 7 cases re-covered in the 12th month, 3 cases were finally diagnosed as deafness (0.89%). Conclusion OAE, ABR combining with BAEP examination may obtain comprehensive diagnosis of hearing impairment for high risk infants, continuous listening comprehension monitoring can effec-tively dynamically observe the hearing impairment, changes and outcome of high risk infants.

Objective To investigate the changes in cellular immune function and biochemical indicators of conventional submarine crew both before and after prolonged deployment at sea.Methods Sixtytwo submariners were recruited as the subjects of our study.Detections were made on T lymphocyte subpopulation and NK cell (CD3 + , CD4 + , CD8 + , CD4 +/CD8 + and NK cells) , heptorenal function as well as 13 biochemical indicators [blood glucose (Glu), aspartate amninotransferase (AST), alanine aminotransferase (ALT), AST/ALT, UREA, creatinine (CREA), potassium (K), sodium (Na) and chlorine (Cl), calcium (Ca), magnesium (Mg), phosphorus (P) and iron (Fe)].Results As compared with those before prolonged deployment at sea, CD3 + levels in the T lymphocyte subpopulation after prolonged deployment at sea (67.02％ ± 7.30％ and 68.91％ ± 8.15％ respectively) , CD4 + levels(33.59％ ± 5.61％ and 36.87％ ± 3.65 ％ respectively) , and CD4 +/CD8 + levels (1.22 ± 0.45, 1.46 ± 0.52 respectively) , were all significantly increased (P ＜-0.05).However, CD8 + levels were obviously decreased(27.32％ ± 8.21％ ,25.25％ ± 4.38％) (P ＜ 0.05).Blood glucose levels were obviously increased [(3.68 ± 0.85) mmol/L and (4.38 ± 1.15) mmol/L respectively] (P ＜ 0.05).Ca levels were also elevated [(2.48 ± 0.11) mmol/L and (2.54 ± 0.12) mmol/L respectively] (P ＜ 0.05).Na levels were respectively [(141.08 ± 5.75) mmol/L,(138.08 ± 2.86)mmol/L] , C1 levels were obviously decreased, which were respectively [(103.12 ± 3.09)mmol/L and(98.89 ±4.28)mmol/L] (P ＜0.01), and Fe levels were also obviously decreased, which were [(23.14 ± 6.75) μmol/L and (21.54 ± 7.34) μmol/L] respectively (P ＜ 0.05).Though biochemical indicators of heptorenal function were elevated to various extents, no statistical differences could be noticed (P ＞ 0.05).Conclusions The immune response of the conventional submariners during this prolonged deployment at sea was enhanced, the blood glucose and Ca levels were elevated, but Fe, Na and Cl levels were decreased, indicating that nutritional components should be adjusted in future submarine deployments at sea,and attention should be paid to the changes in blood glucose, so as to ensure the mental health of the submariners.

Objective To investigate the changes in cellular immune function and biochemical indicators of conventional submarine crew both before and after prolonged deployment at sea.Methods Sixtytwo submariners were recruited as the subjects of our study.Detections were made on T lymphocyte subpopulation and NK cell (CD3 + , CD4 + , CD8 + , CD4 +/CD8 + and NK cells) , heptorenal function as well as 13 biochemical indicators [blood glucose (Glu), aspartate amninotransferase (AST), alanine aminotransferase (ALT), AST/ALT, UREA, creatinine (CREA), potassium (K), sodium (Na) and chlorine (Cl), calcium (Ca), magnesium (Mg), phosphorus (P) and iron (Fe)].Results As compared with those before prolonged deployment at sea, CD3 + levels in the T lymphocyte subpopulation after prolonged deployment at sea (67.02％ ± 7.30％ and 68.91％ ± 8.15％ respectively) , CD4 + levels(33.59％ ± 5.61％ and 36.87％ ± 3.65 ％ respectively) , and CD4 +/CD8 + levels (1.22 ± 0.45, 1.46 ± 0.52 respectively) , were all significantly increased (P ＜-0.05).However, CD8 + levels were obviously decreased(27.32％ ± 8.21％ ,25.25％ ± 4.38％) (P ＜ 0.05).Blood glucose levels were obviously increased [(3.68 ± 0.85) mmol/L and (4.38 ± 1.15) mmol/L respectively] (P ＜ 0.05).Ca levels were also elevated [(2.48 ± 0.11) mmol/L and (2.54 ± 0.12) mmol/L respectively] (P ＜ 0.05).Na levels were respectively [(141.08 ± 5.75) mmol/L,(138.08 ± 2.86)mmol/L] , C1 levels were obviously decreased, which were respectively [(103.12 ± 3.09)mmol/L and(98.89 ±4.28)mmol/L] (P ＜0.01), and Fe levels were also obviously decreased, which were [(23.14 ± 6.75) μmol/L and (21.54 ± 7.34) μmol/L] respectively (P ＜ 0.05).Though biochemical indicators of heptorenal function were elevated to various extents, no statistical differences could be noticed (P ＞ 0.05).Conclusions The immune response of the conventional submariners during this prolonged deployment at sea was enhanced, the blood glucose and Ca levels were elevated, but Fe, Na and Cl levels were decreased, indicating that nutritional components should be adjusted in future submarine deployments at sea,and attention should be paid to the changes in blood glucose, so as to ensure the mental health of the submariners.