酸性微环境下大蒜素增强大鼠自然杀伤细胞的功能活性

Objective To investigate the role of allicin on rat nature killer (NK) cell activity in vitro under acidic microenvironment, and its possible mechanism. Methods CD3- NKR-P1+ NK cells isolated from the rat spleen were cultured in the complete RPMI 1640 medium ( pH 5. 6, pH 6. 5, or pH 7. 2 respectively), and treated with allicin at final concentration of 30 mg/L. Enzyme linked immunosorbent assay (ELISA) was used to determine supernatant interferon (IFN)-γ levels. The percentage of NK cells proliferation and apopotosis was analyzed by flow cytometry. NK cell cytotoxicity toward YAC-1 tumor cells was detected by LDH release assay. Results Proliferation and cytotoxicity of NK cells were significantly suppressed by acidic microenvironment in vitro. Under the cultured condition of acidic pH below 7. 2, allicin seemed to promote NK cells proliferation, which reached to highest level of 33% at pH 5. 6 cultured for 16 h. Correspondingly, at pH of 5. 6, allicin induced a marked increase of IFN-γ concentration in the supernatant from (50. 07 ± 0. 38) (cultured for 4 h) to (64. 59 ± 0. 09) ng/L ( cultured for 16 h). The cytotoxicity of NK cells toward YAC-1 tumor cells was also found strongest under the condition of pH 5. 6 cultured for 16 h. Conclusion Allicin favored to enhance the cytotoxicity of NK cells under the acidic cultured condition, which might be related to the increase of IFN-γ production.     

肾移植术后肝炎性肌纤维母细胞瘤一例

肾移植后发生肝炎性肌纤维母细胞瘤临床罕见。现将本院收治经手术及病理证实的1例报告如下。 患者男,30岁,因“反复右上腹不适半年,体检发现肝尾状叶占位病变1周”入院。患者近半年来感右上腹不适伴疼痛,以钝痛为主,可自行缓解,偶向右肩部放射。入院前1周前于当地医院B超检查发现肝尾状叶占位病变,     

酸性微环境下大蒜素增强大鼠自然杀伤细胞的功能活性

目的 观察体外酸性培养环境下大蒜素对自然杀伤(NK)细胞功能活性的影响,并探讨其机制.方法 以pH 5.6、pH 6.5和pH 7.2的完全RPMI 1640培养基对大鼠脾脏CD3-NKR-P1+NK细胞进行悬浮培养,并给予30 mg/L浓度的大蒜素进行处理.酶联免疫吸附试验(ELISA)检测细胞培养液中干扰素(IFN)-γ的分泌水平,流式细胞仪检测NK细胞的增殖和凋亡率,乳酸脱氢酶法检测NK细胞的功能活性.结果 酸性培养环境下大鼠脾脏NK细胞的增殖能力明显下降,NK细胞功能活性明显受阻.在pH 7.2、pH 6.5和pH 5.6三种不同的培养条件下,大蒜素对NK细胞增殖率的影响表现为随培养环境的pH降低反而升高,以pH 5.6、培养16 h时达最高33.3%.与之相对应,NK细胞分泌的IFN-γ达(64.59±0.09)ng/L,较培养4 h时升高28%,且对小鼠淋巴瘤Yac-1细胞的杀伤活性也达到最高水平.结论 大蒜素可通过提高NK细胞IFN-γ分泌水平明显改善酸性培养环境下NK细胞的功能活性.

Abstract：

Objective To investigate the role of allicin on rat nature killer (NK) cell activity in vitro under acidic microenvironment, and its possible mechanism. Methods CD3- NKR-P1+ NK cells isolated from the rat spleen were cultured in the complete RPMI 1640 medium ( pH 5. 6, pH 6. 5, or pH 7. 2 respectively), and treated with allicin at final concentration of 30 mg/L. Enzyme linked immunosorbent assay (ELISA) was used to determine supernatant interferon (IFN)-γ levels. The percentage of NK cells proliferation and apopotosis was analyzed by flow cytometry. NK cell cytotoxicity toward YAC-1 tumor cells was detected by LDH release assay. Results Proliferation and cytotoxicity of NK cells were significantly suppressed by acidic microenvironment in vitro. Under the cultured condition of acidic pH below 7. 2, allicin seemed to promote NK cells proliferation, which reached to highest level of 33% at pH 5. 6 cultured for 16 h. Correspondingly, at pH of 5. 6, allicin induced a marked increase of IFN-γ concentration in the supernatant from (50. 07 ± 0. 38) (cultured for 4 h) to (64. 59 ± 0. 09) ng/L ( cultured for 16 h). The cytotoxicity of NK cells toward YAC-1 tumor cells was also found strongest under the condition of pH 5. 6 cultured for 16 h. Conclusion Allicin favored to enhance the cytotoxicity of NK cells under the acidic cultured condition, which might be related to the increase of IFN-γ production.     

腹腔镜辅助联合肝脏离断和门静脉结扎的二步肝切除术治疗原发性肝癌的安全性和疗效评价

目的探讨腹腔镜辅助联合肝脏离断和门静脉结扎的二步肝切除术（associating liver partition and portal vein ligation for staged hepatectomy , ALPPS ）治疗原发性肝癌的安全性和疗效。方法回顾性分析2013年12月本院收治的一例巨块型肝癌患者以腹腔镜辅助ALPPS方法切除肿瘤的临床资料。第1步在腹腔镜下结扎门静脉右支并离断左肝内外叶;第2步在第1次术后第8天,评估剩余肝脏体积能满足机体要求后行右半肝的扩大切除术,同时对患者围术期的临床指标和随访资料进行分析。结果第1次手术出血50 ml,手术时长225分钟。第2次手术出血700 ml,手术时长335分钟。切除右肝三叶大小约24．0 cm×22．0 cm×15．0 cm,重量2．7 kg,肿瘤大小约18．0 cm×16．0 cm×14．0 cm,残余肝大小约15．0 cm×13．0 cm×8．0 cm。术后病理结果为原发性肝细胞癌。术后患者出现胆漏,经保守治疗后治愈。术后随访2个月,残余肝再生体积达约1200 ml,AFP降至150 ng／ml,未发现肿瘤复发转移。术后随访至32个月,再次评估残余肝再生体积达约1300 ml,AFP 16 ng／ml,未发现肿瘤复发转移,无疾病生存期达32个月。结论腹腔镜辅助ALPPS手术可作为残肝体积不足的一些较晚期肝癌患者的治疗选择。     

大蒜素对大鼠NK细胞肿瘤杀伤活性的影响

目的: 研究大蒜素对体外培养的大鼠NK细胞肿瘤杀伤活性的影响,并初步分析其可能机制。方法: 免疫磁珠分选法分离大鼠脾脏NK细胞,流式细胞仪检测不同浓度的大蒜素对NK细胞的增殖和凋亡的影响,ELISA检测大鼠脾脏NK细胞IFN-γ的分泌水平,乳酸脱氢酶(LDH)法检测大鼠脾脏NK细胞对小鼠淋巴瘤Yac-1细胞的杀伤活性。结果: 大蒜素对体外培养的NK细胞有明显的促进增殖和抑制NK细胞自然凋亡的作用,并能提高NK细胞分泌IFN-γ的水平和增强对Yac-1细胞的杀伤毒性,且在一定的浓度范围内呈剂量依赖性,其中30 mg/L是大蒜素的相对适宜浓度。结论: 大蒜素可能通过上调NK细胞分泌IFN-γ的水平,增强其体外肿瘤杀伤能力。     

供肝NK细胞对肝移植免疫反应中NF-κB 活性和IL-15表达的影响

 目的：观察大鼠移植肝组织内和外周血白细胞介素15（IL-15）的表达水平及脾组织内核因子κB(NF-κB)的表达活性变化,探讨供肝NK细胞减轻肝移植术后急性排斥反应的机制。方法：Lewis大鼠为肝移植供体,BN大鼠为受体,改良“二袖套”法建立大鼠肝移植模型。应用［60Co］射线照射清除供体免疫细胞和经门静脉回输供肝NK细胞等技术,实验设立3组。A组: 急性排斥组;B组: 单纯［60Co］照射排斥组;C组: ［60Co］照射+供肝NK细胞门静脉回输排斥组。移植术后第1、3、7天收集受鼠外周血清,ELISA法检测IL-15分泌水平,同时切取移植肝行病理学检查,Western blotting法检测移植肝组织IL-15蛋白的表达,凝胶电泳迁移实验（EMSA）检测受鼠脾组织NF-κB表达活性。另外每组各设亚组观察移植后大鼠自然生存时间。结果：B组术后大鼠自然存活时间较A、C组明显缩短,术后发生急性排斥反应程度较A、C组严重;术后第3、7天,3组受鼠外周血IL-15水平均较第1天时显著升高;移植术后第3、7天,B组大鼠血清IL-15 浓度均显著高于A、C组;术后第3、7天,移植肝组织中B组IL-15表达也显著高于A、C组,受鼠脾组织中B组NF-κB表达活性亦显著高于A、C组。结论：IL-15可能作为肝移植急性排斥反应的监测指标,对急性排斥反应的早期诊断和移植物的预后估计具有临床指导意义;供肝NK细胞可能通过下调NF-κB的表达活性来调节IL-15的水平,进而减轻肝移植术后急性排斥反应。     

酸性微环境下大蒜素增强大鼠自然杀伤细胞的功能活性

Objective To investigate the role of allicin on rat nature killer (NK) cell activity in vitro under acidic microenvironment, and its possible mechanism. Methods CD3- NKR-P1+ NK cells isolated from the rat spleen were cultured in the complete RPMI 1640 medium ( pH 5. 6, pH 6. 5, or pH 7. 2 respectively), and treated with allicin at final concentration of 30 mg/L. Enzyme linked immunosorbent assay (ELISA) was used to determine supernatant interferon (IFN)-γ levels. The percentage of NK cells proliferation and apopotosis was analyzed by flow cytometry. NK cell cytotoxicity toward YAC-1 tumor cells was detected by LDH release assay. Results Proliferation and cytotoxicity of NK cells were significantly suppressed by acidic microenvironment in vitro. Under the cultured condition of acidic pH below 7. 2, allicin seemed to promote NK cells proliferation, which reached to highest level of 33% at pH 5. 6 cultured for 16 h. Correspondingly, at pH of 5. 6, allicin induced a marked increase of IFN-γ concentration in the supernatant from (50. 07 ± 0. 38) (cultured for 4 h) to (64. 59 ± 0. 09) ng/L ( cultured for 16 h). The cytotoxicity of NK cells toward YAC-1 tumor cells was also found strongest under the condition of pH 5. 6 cultured for 16 h. Conclusion Allicin favored to enhance the cytotoxicity of NK cells under the acidic cultured condition, which might be related to the increase of IFN-γ production.     

胆囊切除术患者胆汁培养和药敏分析

目的 观察现阶段本中心胆囊切除术患者胆汁病原菌培养阳性率、菌株分布特征和对抗生素 的耐药性,以指导目前抗菌药物在胆道疾病中的合理应用。方法 选取2016年4月至2018年9月湛江市肿瘤医院普通外科诊断为胆囊结石伴急性胆囊炎、胆囊结石伴慢性胆囊炎、胆囊息肉伴慢性胆囊炎的患者 137例,行胆囊切除术,围手术期使用的抗生素主要为头孢孟多酯、头孢唑肟、头孢哌酮/舒巴坦+甲硝唑。 术后均留取胆汁标本,行胆汁细菌、厌氧菌和真菌培养,对病原菌培养和药敏结果进行分析。结果 137 例胆汁标本中62例培养出病原菌,阳性率45.3%,62株病原菌中革兰氏阴性菌22株（35.5%）,革兰氏阳性 菌37株（59.7%）,真菌 3株（4.8%）。排在第一位的菌株仍为大肠埃希菌。革兰阴性菌前2位为大肠埃希菌（10 株,16.2%）和肺炎克雷白杆菌（4株,6.5%）,革兰氏阳性菌前2位为粪肠球菌（9株,14.5%）和溶血葡萄球 菌（9株,14.5%）。药敏结果耐药性分析：革兰阴性菌对亚胺培南、美罗培南和哌拉西林/他唑巴坦的耐药 率最低（均为0）,其次为阿米卡星（4.7%）和头孢哌酮/舒巴坦（9.5%）。革兰氏阳性菌对万古霉素、利奈唑胺和阿米卡星耐药率最低（均为0）,其次为克林霉素（3.0%）、替考拉宁（3.1%）和左氧氟沙星（4.5%）。真菌对常见的抗真菌药两性霉素、氟康唑、伊曲康唑和伏立康唑未见有耐药。137例患者中有22例是胆囊息肉伴慢性胆囊炎,培养出病原菌7株,阳性率为31.8%。结论 本研究显示,胆囊切除术患者胆汁中的主要病原菌仍是大肠埃希菌,但胆汁培养中革兰氏阳性菌株占多数,可能原因为头孢类抗生素使用后革兰 氏阴性菌生长受到抑制以及革兰氏阳性菌感染近年来有上升趋势。胆囊结石伴急性胆囊炎患者术后可根 据临床经验选择抗生素治疗,而无手术条件患者行保守治疗首选哌拉西林/他唑巴坦或头孢哌酮/舒巴坦+ 甲硝唑,如果效果不佳可联用或改用克林霉素、万古霉素,或升级为亚胺培南、美罗培南治疗。胆囊息肉 伴慢性胆囊炎患者胆汁培养菌株阳性率较高,无症状胆囊切除术患者围手术期仍需使用抗生素。     

经皮胆囊穿刺引流术在高龄高危急性胆囊炎患者中的应用

目的探讨胆囊穿刺引流术治疗高龄高危型急性胆囊炎患者的临床疗效。方法选取56例惠有高危型急性胆囊炎的高龄患者，按照就诊顺序随机均分为对照组与研究组各28例，对照组采用腹腔镜下胆囊切除术进行治疗，研究组则采用胆囊穿刺引流术进行治疗，对两组患者的临床疗效进行对照比较。结果研究组的临床疗效明显优于对照组，比较差异具有统计学意义（P〈0．05）。结论采用胆囊穿刺引流术治疗高龄高危型急性胆囊炎，手术成功率较高，临床疗效显著，能够有效减少患者的病死率及并发症发生率，值得临床应用及推广。     

“三吻合器法”在远端胃癌根治术胃肠道重建毕Ⅱ式吻合中的应用

目的探讨在远端胃癌根治术消化道重建中应用"三吻合器法"毕Ⅱ式吻合的临床疗效。方法采用回顾性分析,选取广东省农垦中心医院普通外科2016年1月至2018年12月间收治的远端胃癌76例,按消化道重建方式分为研究组40例(三吻合器法改良毕Ⅱ式),对照组36例(传统毕Ⅱ式),对两组病人的临床疗效进行对比研究。结果研究组手术重建时间、手术重建出血量均少于对照组(P0.05),研究组残胃保有率和预后营养指数高于对照组(P0.05),两组术后病理切缘阴性率比较差异无统计学意义(P0.05)。两组术后均未发生吻合口瘘、吻合口狭窄和吻合口出血等并发症。术后6个月,两组延迟性排空障碍和残胃炎发生率比较差异无统计学意义(P0.05)。但倾倒综合征发生率研究组比对照组低(P0.05)。结论 "三吻合器法"毕Ⅱ式吻合在远端胃癌根治术消化道重建中具有创伤小、手术时间缩短等优点,而且可以提高残胃保有率,减少倾倒综合征的发生,改善术后营养状况,且不降低切缘阴性率,临床效果满意,值得推广。