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.
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 study the prokaryotic expression of the anti-tumor associated glycoprotein-72 (TAG-72) singlechain fragment variable (scFv) antibody and its specific affinity to hepatocellular carcinoma cell lines and tissues. Methods The cDNA of anti-TAG-72 scFv antibody was inserted into pCANTAB5E to obtain phage vector anti-TAG-72 -scFv-pCANTAB5E. Isopropyl-β-D-thiogalactoside (IPTG) was used to induce the expression of anti-TAG-72 scFv antibody. SDS-PAGE and Western blot were used to identify the anti-TAG-72 scFv antibody. Human hepatocellular carcinoma cells were cultured, and TAG-72 was determined with the obtained scFv by immunohistochemistry in the cells and paraffin-embedded hepatocellular carcinoma tissues. Results SDS-PAGE and Western blot showed that the anti-TAG-72-scFv antibody was successfully expressed. Anti-TAG-72 scFv could bind to hepatocellular carcinoma cell lines SMMC7721, HepG2, and HHCC, but not to BEL7402, suggesting that SMMC7721, HepG2, and HHCC cells expressed TAG-72. For the 40 cases of hepatocellular carcinoma tissues, the positive rate of TAG-72 in stage Ⅰ and Ⅱ-Ⅲ was 23.08% (3/13) and 62.96%(17/27), respectively. While no TAG-72 expression was found in the 10 normal cases. TAG-72 expression was significantly different between hepatocellular carcinomas and normal tissues (Plt;0.05). Conclusions The prokaryotic expression of anti-TAG-72 scFv antibody is successfully achieved, and can be used to identify TAG-72 antigen. TAG-72 is highly expressed in hepatocellular carcinoma, but not in normal liver tissue, which may be suggested as cancer marker of hepatocellular carcinoma.
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.
Objective To evaluate the analgesic effect of intra-articular ropivacaine with lidocaine. Methods A double-blind randomized controlled trial was conducted. Ninety patients receiving selective knee arthroscopy were randomized into three groups of 30 patients. At the end of the operation, before the release of the tourniquet, an intra-articular injection was administered to each patient through arthroscope, in accordance with their random allocation: 0.9% normal saline (normal saline group); 100 mg ropivacaine (ropivacaine alone group) and 100 mg ropivacaine and 100 mg 2% lidocaine (ropivacaine with lidocaine group). Pain intensity was assessed after the operation using the 100-mm visual-analog scale (VAS), and the amount of supplemental analgesics used within the following 24 hours were recorded. Results The VAS scores of 2 hours postoperatively at rest, and 1, 2, 4, and 8 hours postoperatively at motion, were significantly higher in the normal saline group than in ropivacaine alone group (Plt;0.05). The VAS scores 0.5, 1 and 2 hours postoperatively at rest, and at the awaking moment, 0.5, 1, 4, 8, and 24 hours postoperatively at motion, were significantly higher in the normal saline group than in ropivacaine with lidocaine group (Plt;0.05). Conclusion Intra-articular ropivacaine can reduce a patient’s pain after operation. The combination of lidocaine with intra-articular ropivacaine can reduce the patient’s pain severity immediately after the operation and achieve an early analgesic effect.
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.
