专题论著: 新型冠状病毒肺炎
目的 探讨新冠疫情影响下日间手术制度在乳腺外科住院模式中的应用情况。方法 2021年1月—12月广州市第一人民医院实施日间手术制度,对乳腺外科30张床位实施预约管理,依据患者病情进行日间手术预约,合理安排患者入院。结果 2021年1月—12月我院乳腺外科共入院2 787人/次,手术人数1 497人,其中日间手术720台,占比48.10%。结论 日间手术制度可提升了医院手术服务能力,增加手术量,缩短了平均住院日,为患者提供方便、高效的医疗服务,值得基层医院推广。
论著
目的 探究小分子化合物逆转素(reversine,Rev)对胆管结扎(BDL)诱导的大鼠胆汁淤积性肝损害、纤维化、上皮细胞-间充质转化以及胆管反应的影响。方法 雄性Lewis大鼠随机分成三组,每组各5只。按照如下处理:BDL组大鼠行2周的胆管结扎;BDL+Rev组行胆管结扎同时给予腹腔注射逆转素;对照采用假手术(Sham)。2周后获取血液和肝组织。血指标检测总白蛋白(TP)、总胆红素(TBIL)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ谷氨酰转肽酶(GGT)。H&E染色检测肝组织病理。Azan染色检测组织胶原蛋白。免疫组化检测肝组织α平滑肌肌动蛋白(α-SMA)、结蛋白(Desmin)、波形蛋白(Vimentin)、细胞角蛋白(CK7,CK19)、β-连环蛋白(β-Catenin)以及上皮细胞粘附分子(EpCAM)蛋白的表达情况。结果 胆管结扎导致肝脏合成的总白蛋白量下降,总胆红素(TBIL)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ谷氨酰转肽酶(GGT)水平明显上升,逆转素处理使下降的总白蛋白上升,使上升的总胆红素(TBIL)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、γ谷氨酰转肽酶(GGT)水平向正常水平回复。逆转素可以缓解胆汁淤积引起的肝纤维化,表现为下调BDL引起的胶原蛋白和α-SMA蛋白沉积。逆转素可以抑制胆汁淤积引起的上皮细胞-间充质转化表现为逆转素明显降低BDL导致的Desmin和Vimentin的表达。逆转素可以抑制胆汁淤积引起的胆管反应表现为明显减少CK7和CK19阳性胆管的表达含量。逆转素抑制胆汁淤积引起的胆管反应与调节β-Catenin和EpCAM的表达有关。结论 逆转素可以缓解胆汁淤积引起的大鼠肝损害,具有一定的保护作用。逆转素可以成为一种潜在治疗药物。
Objective To investigate the effect of reversine (REV) on bile duct ligation (BDL) -induced hepatic damage, fibrosis, epithelial-mesenchymal transformation, and ductular reaction in rats. Methods Male Lewis rats were randomly divided into three groups with 5 rats in each group. Bile duct ligation was performed in the BDL group for two weeks. BDL+ REV group was treated with bile duct ligation and intraperitoneal injection of reversine. The control group was Sham operation (Sham). Blood and liver tissue were obtained after 2 weeks. Blood indexes were determined for total albumin, total bilirubin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and γ-glutamyl transpeptidase. Hepatic histopathology was detected by H&E staining. Azan staining was used to detect tissue collagen deposition. Immunohistochemistry was used to detect the expression of α-SMA, desmin, vimentin, cytokeratin, β-catenin and epithelial cell adhesion molecule (EpCAM) protein. Results Bile duct ligation resulted in the decrease of total albumin synthesis in liver, and the increase of total bilirubin, glutamic-oxalacetic transaminase, alkaline phosphatase and γ-glutamyltranspeptidase. The levels of total bilirubin, aspartate transaminase, alkaline phosphatase and γ-glutamyltranspeptidase returned to the normal level with reversine treatment. Reversine could alleviate cholestasis-induced liver fibrosis by downregulating BDL-induced deposition of collagen and α-SMA protein. Reversine inhibited cholestasis-induced epithelial-mesenchymal transformation by significantly reducing BDL-induced desmin and vimentin expression. Reversine could inhibit cholestasis-induced ductular reaction by significantly reducing the expression of CK7 and CK19 positive biliary cells. Inhibition of cholestasis induced ductular reaction by reversine was associated with regulation of β-catenin and EpCAM expression. Conclusion Reversine can alleviate liver damage caused by cholestasis in rats and have a protective effect. Reversine may be a potential treatment that need further investigation.
临床诊疗
目的 从生物力学应力调控角度,探讨组织间质液压升高诱发肝纤维化的分子细胞机制。方法 人肝星状细胞随机分为3组,模型组:将其置于压力箱中施加恒定的高于大气压50 mmHg压力;实验组:加入ROCK抑制剂Y-27632(浓度10 μmol)置于与实验组相同条件;对照组:不加压置于相同培养箱中。应用RC-PCR检测其α-SMA、RhoA、ROCK mRNA的表达量,并应用免疫荧光染色分析其α-SMA、ROCK1、ROCK2蛋白表达情况。结果 间质液压变化对人肝星状细胞中α-SMAmRNA的表达量比对照组表达量增加,RhoA mRNA的表达先上升后下降,只对早期压力有变化,而 ROCK mRNA的表达量无明显变化。抑制剂组和压力组α-SMA 、ROCK1荧光强度较对照组增强,ROCK2荧光强度无明显变化,其中模型变化更显著。结论 细胞间质液压升高能通过RhoA/ROCK1信号通路引起肝星状细胞的活化。
临床诊疗
目的 探讨入院准备中心制度在我院乳腺外科住院预约统筹管理的实践情况。方法 2017年1月—12月广州市第一人民医院入院准备中心对乳腺外科30张床位实施病床集中预约管理,依据患者病情进行预约,合理安排患者入院。结果 2017年1至12月我院乳腺外科共预约入院1225人/次,成功办理预入院1096人/次,约占乳腺外科总入院人数的77.85%。预入院进行手术患者415人/次,其中3日内手术患者为285人/次,预入院三日内手术率为68.67%。结论 预入院制度对乳腺外科病床进行集中预约管理能有效保证床位充分使用,为患者提供方便、有效的医疗服务,值得基层医院推广。
临床诊疗
目的 探讨Reversine对人肝星状细胞系LX-2凋亡的影响。方法 设对照组和Reversine干预组,其中Reversine干预组分为7个浓度,分别为1,5,10,20,40,80,120 μg/mL,CCK-8法检测Reversine对LX-2增殖的影响,选取最佳浓度。将细胞重悬在加入5 μL FITC-Annexin V和5 μL PI,用流式细胞仪进行了凋亡率分析,免疫荧光检测凋亡蛋白bcl-2及caspase 3。结果 Reversine可促进LX-2细胞凋亡,随着Reversine浓度增加,LX-2的凋亡可呈剂量依赖关系,其中10 μg/mL为最佳浓度,LX-2细胞的bcl-2蛋白的表达显著下降而cleaved-caspase 3的表达显著上升。结论 Reversine可通过促进caspase-3蛋白活化、抑制bcl-2蛋白表达的方式诱导LX-2凋亡。
临床诊疗
目的 探讨腹腔镜手术与传统开腹手术对右半结肠癌患者免疫功能指标及临床疗效的影响。方法 选取80例右半结肠癌患者,随机分成两组,各40例分别行腹腔镜手术及传统开腹手术,分别比较两组患者手术前后免疫功能指标及相关临床疗效的差异。结果 腹腔镜组及开腹组术后免疫指标CD3+、CD4+、CD8+均出现不同程度下降,但腹腔镜组免疫指标CD3+、CD4+均优于开腹组(P<0.05);腹腔镜组术中出血量、术后胃肠排气功能恢复时间、住院时间均优于开腹组(P<0.05);两组手术时间及淋巴结清扫个数方面无显著差异(P>0.05)。结论 腹腔镜手术治疗右半结肠癌不仅对患者免疫功能损害较低,还较开腹术具有术中出血量少、术后恢复快等优点,值得临床推广。
论著
目的 构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型。方法 采用低浓度加量持续诱导法,诱导吉西他滨耐药乳腺癌细胞4T1耐药株,命名为4T1/Gem;CCK-8法测定4T1与4T1/Gem细胞的增殖抑制率,计算耐药指数; Western blot法检测细胞P-gp蛋白表达;B超引导下注射4T1/Gem细胞悬液诱导裸鼠肝脏成瘤;HE染色观察肿瘤组织病理情况,免疫组化法检测瘤组织ER、PR、HER2、Ki-67和P-gp蛋白的表达。结果 经过14个月的诱导成功建立4T1/Gem细胞株,可在含40 μg/mL的Gem培养液中稳定生长。4T1/Gem细胞耐药指数为4T1细胞的788.547倍。与亲代相比,4T1/Gem处于G1期和G2期的细胞增加,S期细胞减少;上调P-gp蛋白的表达。4T1/Gem细胞成功建立裸鼠乳腺癌肝转移模型,瘤组织中ER、PR、HER2蛋白阴性表达,Ki-67阳性10%和P-gp蛋白阳性表达。结论 成功构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型,为开发治疗乳腺癌肝转移化疗耐药的药物提供实验基础。
Objective To construct a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem) and establish a nude mouse model of breast cancer with hepatic metastatic. Methods A gemcitabine-resistant variant of the breast cancer 4T1 cell line was induced by gradually increasing the concentration of gemcitabine; this variant is referred to in this study as 4T1/Gem. The proliferation suppression rates of 4T1 and 4T1/Gem cells were determined by using the CCK-8 essay to evaluate the drug resistance indices of the cell lines. Western blot analysis was used to detect P-gp protein expression. Under ultrasonography, a 4T1/Gem cell suspension was injected into nude mice to induce liver tumors. H&E staining was used to observe tumor pathology, and immunohistochemistry was used to detect the expression of ER, PR, HER-2, Ki-67, and P-gp. Results After 14 months of induction, a 4T1/Gem cell line is established successfully. The cell line can grow stably in culture liquid containing 40 μg/ml gemcitabine. The drug resistance index of 4T1/Gem is 788.547. Compared with the 4T1 cell line, the 4T1/Gem cell line can upregulate P-gp protein expression and successfully establish a nude mouse model of breast cancer with hepatic metastatic. ER, PR, and HER-2 proteins exhibit negative expression in the tumor tissue. The positive expression of P-gp and 10% of Ki-67 proteins is also observed. Conclusion This study successfully constructs a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem)and establishes a nude mouse model of breast cancer with hepatic metastatic, thereby providing an experimental basis for developing and treating a drug-resistant variant of breast cancer.
论著
目的 探讨生理情况下高龄肝脏与低龄肝脏的区别,寻找可以区分高龄肝脏与低龄肝脏的指标。方法 取6周龄SD大鼠和9月龄以上退役SD大鼠各5只,采用超声弹性成像检测肝脏硬度、全自动生化检测仪检测血清学指标、H&E染色观察肝脏形态结构、Sirius Red染色及Masson染色检测胶原纤维的沉积、免疫组化SP法检测TGF-β1、p16INK4a、SMP-30蛋白的表达。结果 高龄组和低龄组之间血清学指标、胶原纤维沉积及TGF-β蛋白、p16INK4a蛋白的表达无差异;超声弹性成像检测低龄组Vs值为(1.21±0.09)m/s,高龄组为(1.32±0.05)m/s(P=0.033);SMP-30蛋白低龄组IOD值为138244.988±51286.257,高龄组为116240.170±35017.936(P=0.007)。结论 高龄大鼠与低龄大鼠肝脏的硬度及SMP-30蛋白的表达存在差异,随着年龄的增加肝脏硬度增大,SMP-30蛋白表达下降。肝脏硬度与SMP-30蛋白可作为区分高龄肝脏与低龄肝脏的指标。
Objective To investigate the differences between elderly and younger liver. Methods In accordance with the age of the SD rats into two groups: younger group (Group Y, 6 weeks, n=5) and elderly group (Group O, 40 weeks or more, n=5). Data were compared by using ultrasound elasticity imaging to detect liver stiffness, automatic biochemical detector to gauge serum indexes, H&E staining to observe the liver morphological structure, Sirius Red staining and Masson staining to assay the collagen fibers deposition, Immunohistochemistry to identify the expression of TGF-β1, p16INK4a and SMP-30 protein. Results Serum indexes, collagen deposition, TGF-β1 and p16INK4a protein expression were no statistically significant difference between two groups. The Vs value was (1.21±0.09) m/s in Group Y and (1.32±0.05) m/s in Group O (P=0.033). and the IOD value of SMP-30 protein between Group Y and Group O were 138244.988±51286.257 and 116240.170±35017.936 (P=0.007). Conclusion The degree of liver stiffnessnd and SMP-30 protein in elderly and younger liver are different.Increased the degree of liver stiffness and decreased the expression of SMP-30 protein in the elderly SD rats. Liver stiffness and SMP-30 protein could be used as indicators to distinguish between elderly and younger liver.
临床诊疗
目的 动态观察乳腺癌患者辅助内分泌治疗5年后的血脂及肝功能水平的变化,探求辅助内分泌治疗与高脂血症及脂肪肝发病率的关系。方法 56例乳腺癌患者实行辅助内分泌治疗,术后随访5年动态抽血测定其总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL)、高密度脂蛋白胆固醇(HDL)及谷草转氨酶(AST)、谷丙转氨酶(ALT)、直接胆红素(DBIL)、总胆红素(TBIL)等参数的变化,B超监测其肝脏变化。结果 经过2年内分泌治疗TG由(1.203±0.723)mmol/L上升至(1.701±1.271)mmol/L,5年内分泌治疗后TG降至(1.389±0.706)mmol/L。经过2年内分泌治疗LDL由(2.497±0.990)mmol/L上升至(2.950±0.984)mmol/L,5年内分泌治疗后LDL为(2.867±0.886)mmol/L。结论 辅助内分泌治疗2年会导致其TG和LDL的升高,5年随访仅发现LDL升高,辅助内分泌治疗会增加乳腺癌患者诱发心血管疾病的风险。