Objective
To study effect of carcinoembryonic antigen (CEA) positive targeted lymphocytes on gastric cancer cells in vitro and in vivo.
Methods
The peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers. The recombinant vector anti-CEA-scFv-CD3ζ-pcDNA3.0 was transfected into the PBMCs by lipofectamine 2000, by this means, the CEA special lymphocytes were obtained. Meanwhile, the PBMCs transfected with empty plasmid pcDNA3.0 were used as control (empty vector lymphocytes). The different lymphocytes and gastric cancer cells (CEA positive KATOⅢ gastric cancer cells and CEA negative BGC-823 gastric cancer cells) were co-cultured, then the ability to identify the gastric cancer cells and it’s effect on apoptosis of gastric cancer cells were observed at 24 h or 36 h later respectively. The CEA special lymphocytes and empty vector lymphocytes were injected by the tail vein of nude mice bearing gastric cancer cells, then it’s effect on the tumor was observed.
Results
① The CEA special lymphocytes could strongly identify the KATOⅢ gastric cancer cells (identification rate was 72.3%), which could weakly identify the BGC-823 gastric cancer cells (identification rate was 7.8%). ② The apoptosis rate of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly higher than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (P=0.032), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (P=0.118). ③ The tumor volume of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly smaller than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (F=5.010, P<0.01) or the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells (F=4.982, P<0.01), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (F=1.210, P>0.05).
Conclusion
CEA special lymphocytes can promote cell apoptosis and inhabit tumor reproduction of CEA positive gastric cancer cells in vitro and in vivo.
Methods To explore the level of delirium knowledge of geriatric nurses in Sichuan province and analyze the factors, so as to provide the basis for systematic and targeted knowledge training on delirium and clinical management. Methods Using the self-designed “the Questionnaire of Elderly Delirium Knowledge”, geriatric nurses from 22 hospitals in Sichuan province were investigated through a convenient sampling method from September 2018 to February 2019. Results A total of 475 geriatric nurses were investigated. The average delirium knowledge score of the 475 geriatric nurses was 69.51±12.42. Multiple linear regression analysis showed that the main factors affecting the score of delirium-related knowledge were the education of nurses (P=0.037), technical title (P<0.001), years of working in the geriatric department (P=0.001), and the level of working hospital (P=0.001). Conclusions The level of delirium knowledge of geriatric nurses is low and can not meet the needs of clinical work. Nursing managers should carry out delirium knowledge training according to the different characteristics of nurses.
ObjectiveTo investigate the method for generating anchor chemric T lymphocytes that can target tumor associated glycoprotein-72 (TAG72) antigen and analyze their repressive effects on proliferation of TAG72 positive hepatocarcinoma cells. MethodsFirstly, peripheral blood mononuclear cells (PBMCs) from healthy volunteers were isolated. And then, CD8+ T cells were isolated from PBMCs via magnetic activated cell sorting (MACS). These lymphocytes were transfected with recombinant vector, anti-TAG72-scFv-CD28-pcDNA3, through Lipofectamine2000 to gernerate anchor chimeric TAG72-specific CD8+ T cells. SMMC7721 (TAG72 positive) hepatocarcinoma cells were co-cultured with chimeric T lymphocytes and their cell cycles were analyzed by flow cytometry (FCM). ResultsAnchor chmeric T lymphcytes targetting TAG72 recognized TAG72 positive SMM7721 cells and repressive effects on their proliferation were observed by flow cytometry. ConclusionAnchor chmeric T lymphcytes targetting TAG72 on tumor surface can specifically recognize TAG72 positive hepatocarcinoma cells and may exert repressive effect on their proliferation.
Objective To construct a green fluorescent protein expression plasmid pEGFP-C3-anti-TAG72 scFv-CD28, containing anti-TAG72 single chain variable fragment (scFv) fused into the transmembrane and intracellular domain of the signal-transducing chain of CD28 gene, and to transfect it into peripheral blood mononuclear cells． Methods Recombinant transmembrane and intracellular domain of CD28 cDNA and anti-TAG72 scFv cDNA fragment was subcloned into pEGFP-C3 vector. Recombinant clones were selected by Kanamyein， and then identified by PCR， enzyme digestion analysis and DNA sequencing. The recombinant plasmid was transfected into peripheral blood mononuclear cells by means of lipofection. The recombinant protein expression was confirmed by immunocytochemistry, laser scanning confocal microscope, PCR and Western blot analysis. Results The fused gene fragment of anti-TAG72 scFv-CD28 was successfully inserted into pEGFP-C3 plasmid, and it was confirmed by enzyme digestion and DNA sequencing. The fused anti-TAG72 scFv-CD28 gene and its protein was identified in peripheral blood mononuclear cells． Conclusion The eukaryotic expression plasmid pEGFP-C3-anti-TAG72 scFv-CD28 was successfully constructed and transiently expressed in peripheral blood mononuclear cells， which would lay a foundation for further studies on the role of it to activate tumor-associated antigen-specific T lymphocyte, for generating of modified T lymphocytes targeting gastrointestinal tumors.
Objective To evaluate the effect of day surgery with diagnosis-related groups (DRG) evaluation indicators. Methods The inpatients undergoing surgery in Beijing Tongren Hospital of Capital Medical University between March and September 2022 were enrolled in this study. The medical quality, medical efficiency, hospitalization cost, DRG insurance payment and other DRD-related indicators were retrospectively collected and compared between day surgery patients and non-day surgery patients, and the average length of hospital stay and hospitalization costs were compared between the two surgery modes within DRGs. Chi-square test was used for enumeration data, and t-test and Mann-Whitney U test were used for measurement data. Results A total of 29339 day surgery patients and 19019 non-day surgery patients were enrolled. In the day surgery group, the proportions of local patients (71.71% vs. 68.62%), routine discharge (99.93% vs. 99.78%), and class A incisions (99.92% vs. 99.55%) were better than those in the non-day surgery group (P<0.05), and the average length of hospital stay [(1.00±0.00) vs. (6.98±5.00) d] and the average hospitalization costs [(7306.62±4605.73) vs. (24913.97±24623.54) yuan] were lower than those in the non-day surgery group (P<0.05). The top 2 reduction of average length of hospital stay were in the CB49 group and CB39 group, decreasing by 87.45% and 86.24%, respectively. The top 2 reduction of hospitalization costs were in the DC19 group and CC15 group, decreasing by 84.15% and 73.61%, respectively. DRG payment balance of medical insurance in the day surgery group was higher than that in the non-day surgery group (22.95% vs. 5.98%). Conclusions Day surgery not only ensure the medical quality, but also shorten the length of hospital stay and reduce the burden of medical expenses. Day surgery can effectively improve the utilization efficiency of hospital bed resources, it is an effective measure to promote the high quality development of hospital and comply with DRG payment reform.
Objective To discuss left ventricular reconstruction methods and effectivness in patients with left ventricular aneurysm after myocardial infarction. Methods Between June 2003 and August 2008, 23 patients with left ventricular aneurysm after myocardial infarction were treated. Of them, 13 were male and 10 were female with an average age of 61.2 years (range, 47-74 years). According to New York Heart Association (NYHA) criteria for cardiac function, there were 3 cases of grade I, 6 cases of grade II, 10 cases of grade III, and 4 cases of grade IV. The coronary arteriongraphy showed singlevesseldisease in 2 cases, double-vessel disease in 5 cases, triple-vessel disease in 16 cases. The locations of ventricular aneurysm were the apex cordis in 18 cases, antetheca and parieslateral is in 4 cases, and interior wall in 1 case. The left ventricular ejection fraction was 36.52%± 12.15%, and left ventricular diastol ic final diameter was (62.30 ± 6.52 ) mm. Nine patients received standard l inear repair, 6 patients received standard l inear repair after endocardial ring shrinkage, and 8 patients received patch suture after endocardial ring shinkage. Results Two cases died perioperatively, and re-thoracotomy was performed to stop bleeding in 1 case. Incisions healed by first intention in the other patients without early compl ication. Twentyone patients were followed up 7-48 months (median, 19 months). At 6 months after surgery, the left ventricular ejection fraction 46.52% ± 9.41% were significanly improved when compared with that at preoperation (t=2.240, P=0.023); the left ventricular diastol ic final diameter (52.23 ± 5.11) mm were significantly decreased when compared with that at preoperation (t=2.170, P=0.035). The cardiac function according to NYHA criteria was at grade I in 8 cases and at grade II in 13 cases. One patient died of cerebral hemorrhage at 18 months after operation and the activities of daily l iving recoverd in the others. Conclusion Individual theraputic methods are used according to patients’ different conditions for left ventricular aneurysm after myocardial infarction.
Objective To investigate the effect of tumor associated glycoprotein-72 （TAG-72） redirected T lymphocytes on breast cancer cells. Methods Peripheral blood mononuclear cells （PBMCs）were isolated from healthy volunteers. The recombinant vector anti-TAG-72-scFv-CD3ζ-pcDNA 3.0 were transfected into PBMCs by lipofectamineTM2000 （transfection group）， PBMCs transfected with plasmid pcDNA 3.0 as control group. MCF-7 and Bcap37 cells were cocultivated with PBMCs of transfection group and control group， respectively， and antitumor response of G1 block was observed. Results G1 block rate of MCF-7 cells in transfection group was （82.3±6.9）%， which was significantly higher than that in control group 〔（43.4±3.9）%， P＜0.05〕. G1 block rate of Bcap37 cells in transfection group was （51.3±4.7）%， and not differed from that in control group 〔（45.6±2.5）%， P＞0.05〕. Conclusion TAG-72 redirected T lymphocytes can inhibit the cell proliferation of TAG-72 positive breast cancer cells， and it may provide valuable tools for the cellular immunotherapy.
