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.
目的 调查安置永久性人工心脏起搏器的老年患者术前焦虑状况，为制订护理对策提供依据。 方法 2004年7月-2008年7月收治需安置永久性人工心脏起搏器的心脏病患者78例，术前采用焦虑自评量表（SAS）对其进行问卷调查，并采用自制一般情况问卷调查了解情况。 结果 植入永久性心脏起搏器的老年患者术前SAS得分高于国内常模（Plt;0.05）。根据自制调查问卷结果，差异有统计学意义（Plt;0.05）的项目:老年患者随年龄增长焦虑量表评分降低；丧偶者焦虑评分高于有配偶者；完全公费、部分公费、自费的焦虑量表评分依次增高；老年患者对起搏器知识了解程度越少，焦虑评分越高；无家人陪伴者比有家人陪伴者焦虑评分高。而性别、文化水平差异无统计学意义。 结论 植入永久性起搏器的老年患者术前多数存在焦虑情绪，且焦虑与年龄、婚姻、费用支付方式、有无家人陪伴、相关知识等有一定关系，医护人员应针对性地做好患者术前护理，帮助患者面对现实，以积极的心态接受手术治疗。
目的:研究替米沙坦对合并高血压的阵发性房颤患者转复为窦性心律后复发的影响。方法: 选择84例合并高血压的阵发性房颤患者,随机分成观察组及对照组,观察组降压药使用替米沙坦,对照组使用氨氯地平,两组均使用胺碘酮复律,随访2年,观察两组患者的疗效及超声心动图(左房内径)变化。结果:观察组治疗合并高血压的阵发性房颤复发率显著低于对照组。结论:替米沙坦联合抗心律失常药物,能有效防止房颤复发,维持窦性心律。
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.
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.
目的 了解成都市卧床老年人的抑郁发生情况及影响因素。 方法 对2009年12月－2011年2月卧床时间＞1个月的325例卧床老年人采用老年抑郁量表、焦虑自评量表、生活满意度指数A进行调查，并对影响的抑郁的相关因素进行统计分析。 结果 成都市卧床老年人抑郁的发生率为57.5%。不同病情、生活自理能力、焦虑情况、社会交往情况、生活的满意度和家庭功能的老年人，其抑郁评分差异有统计学意义（P＜0.05）。多重线性回归分析发现影响老年人抑郁的主要因素有病情、卧床分级、焦虑、社会活动、满意度、文化程度，其中对生活满意、有社会活动、文化程度高是保护因素，而焦虑、病情较重、卧床等级高是抑郁的危险因素。 结论 卧床老年人的抑郁发生率较高，应加强对卧床老年人，特别是病情重、焦虑、大部分或者全天卧床、低文化的卧床老人抑郁发生的关注，鼓励老年人增加社会活动、提高老年人对生活的满意度和增进他们的心理健康。
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.
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 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.