女性性高潮障碍多

资料显示，女性性高潮并非人人尽有，而能够获得性高潮的女性也不可能每次性生活都能达到性高潮，有多种因素会对其产生影响。国外学者统计新婚之时女性达到性高潮者为4％，几年内达到高潮者占50％。我国学者对200名育龄妇女进行调查，发现有50％从未有过性高潮。另一个资料显示，女性在性生活中每次都达到性高潮的仅占10％。     

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.     

大鼠原位肝移植术后胆道并发症的影响因素

目的 研究大鼠原位肝移植术后胆道并发症的各种影响因素,为建立更稳定的大鼠原位肝移植模型提供实验依据.方法 雄性SD大鼠87只,完全随机法分组,其中移植组48只、非移植组36只、假手术组3只,实验共分胆管-胆管重建移植组、胆管-十二指肠重建移植组、胆管-胆管重建+肝动脉结扎非移植组、胆管-胆管重建非移植组、胆管-十二指肠重建+肝动脉结扎非移植组、胆管-十二指肠重建非移植组和假手术组7组.各手术处理组以出现明显胆道并发症或移植后14 d作为观察时间终点.通过肝脏大体标本肉眼观察、病理学检测和血清学分析等判断胆道并发症的发生率及其严重程度并进行各组间比较.结果 移植组总胆道并发症发生率为69.6%,明显高于非移植组(25.0%)和假手术组(0%),差异有统计学意义(均P<0.05).在移植组和非移植组中,胆管-十二指肠重建组的胆道并发症发生率明显高于胆管-胆管重建组(90.9%比50.0%,38.9%比11.1%,均P<0.05).非移植组中,结扎肝动脉组的胆道并发症高于不结扎肝动脉组(38.9%比11.1%,P<0.05).结论 经典"二袖套法"大鼠原位肝移植模型尚不完善,仍需进一步改进.就大鼠原位肝移植模型而言,胆管-十二指肠重建方式并非理想的选择.     

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.     

单人裸眼建立大鼠原位肝移植模型

目的 探讨单人裸眼建立大鼠原位肝移植模型的学习曲线.方法 将作者不同阶段制作该模型的600例手术资料进行归纳总结,比较各阶段的供体时间、冷灌注时间、无肝期、手术成功率、1周存活率、长期存活率以及手术并发症的差异.结果 第Ⅰ阶段各手术操作时间均较后三阶段长,其差异有统计学意义(P＜0.01).手术成功率从第Ⅰ阶段的26.7％逐渐上升至第Ⅳ阶段的接近100.0％,1周存活率以及长期存活率均稳步提高.手术并发症主要为早期的血管并发症以及晚期的胆道并发症.第Ⅱ、Ⅲ、Ⅳ阶段的手术死亡率较第Ⅰ阶段大幅下降,手术并发症的发生率也逐步下降,建立模型的技术逐渐稳定.结论 初学者经过系统的持之以恒的训练学习,可逐渐掌握大鼠原位肝移植模型建立的技术要领.     

Decorin/fusin双基因修饰骨髓间充质干细胞趋化动力学及抗纤维化的研究

Objective To investigate in vitro chemotaxis dynamics and anti-fibrillar synthesis of dual-genes-modified mesenchymal stem cells ( MSCs). Methods A recombinant lentiviral-mediated chimera fusion genes was constructed using pLTR-CMV-DCN, pLTR-CMV-FIB-DCN, pLTR-CMV-FIB-Fusin plasmids and a pLVX-puro vector. Viral particles titer 1×107 cfu/ml obtained through HEK 293 cells packages were transfected into MSCs followed by puromycin selection. Transfection efficiency, gene expression were analyzed;chemotaxis,Transwell in vitro assay of fibrillar synthesis were assessed. Results Transfection efficiency 11%, after selection and purification 30%. Migrational ability and anti-fibrillar synthesis were shown under various conditions. Conclusion Dual-gene-modified mesenchymal stem cells could undergo a conformational structural change under specific microenvironment, affecting its genes expression, chemotaxis dynamics,and in vitro fibrillar synthesis.     

羟基荧光素二醋酸盐琥珀酰亚胺脂标记大鼠肝脏NK细胞体内转输示踪

目的 利用羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)标记大鼠肝脏NK细胞进行体内示踪,观察其在受鼠体内的存活.方法 经SD大鼠门静脉转输1×106个/ml CFSE体外标记新鲜分离的SD供鼠肝脏NK细胞,于输人后1、3、7、15 d分别切取肝脏和脾脏行冰冻切片免疫荧光观察,并收集门静脉血监测淋巴细胞和中性粒细胞的比例变化.结果 CFSE对大鼠肝脏NK细胞的标记率为98.63%;转输NK细胞后第1天受鼠肝脏内CFSE荧光最强,7 d时荧光变微弱,15 d时荧光基本消失;脾脏在第1、3天时可见较弱荧光,7 d时荧光消失.转输NK细胞后相同时间点肝脏内CFSE荧光均明显强于脾脏.转输肝脏NK细胞的受鼠门静脉血中性粒细胞比例第1天时最高,随时间延长逐渐下降,第15天时基本回复正常水平,淋巴细胞比例仅一过性降低.结论 转输的肝脏NK细胞在受体内的数量随时间延长而减少,在受体内的存活时间约为15 d.     

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。     

Vps4在肝癌及癌旁组织中的差异性表达及其临床意义

【摘要】 目的 液泡蛋白分选因子4（vacuolar protein sorting 4 Vps4）是调节细胞分裂、病毒芽胞形成的重要因素,本文检测其在肝细胞癌（HCC）患者的癌旁组织、癌组织及正常肝组织中差异性表达,分析该蛋白与肝癌临床病理资料之间的关系。方法 应用免疫组织化学法检测56对经手术切除并有明确病理诊断为肝细胞癌的癌组织、癌旁组织及11例正常肝组织标本中Vps4的表达。比较上述组织中Vps4的表达并分析该蛋白表达与临床病理资料之间的关系。结果=1）癌组织、癌旁组织、正常肝组织、癌栓、转移性淋巴结等组织的细胞质中均可表达Vps4;２）癌组织中Vps4平均表达量低于癌旁组织和正常肝组织,差异有统计学意义（Ｐ＜０．０５）;正常肝组织与癌旁组织中Vps4的平均表达量无明显差异（Ｐ＞０．０５）;３）Vps4的表达与ＨＢＶ－ＤＮＡ复制量有关,与年龄、性别、肿瘤直径、肝内多发病灶、肝外淋巴结转移、肝胞膜完整性、肝硬化、ＡＦＰ表达量、Ｃｈｉｌｄ－Ｐｕｇｈ分级、脉管癌栓、Edmonson分级、TNM分期等无关。结论 癌组织中Vps4表达明显低于正常肝组织及癌旁组织,且与ＨＢＶ－ＤＮＡ复制量有关,Vps4表达可能影响肝癌的发生和发展。     

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.