富氢生理盐水对兔脊髓损伤神经细胞凋亡及凋亡相关蛋白表达的影响

目的探讨富氢生理盐水腹腔注射对家兔脊髓损伤神经细胞凋亡及凋亡相关蛋白表达的影响。方法选用30只雄性家兔,随机分为3组,即健康组、对照组和治疗组,每组10只。对照组采用对生理盐水对脊髓损伤家兔进行治疗,治疗组采用富氢生理盐水治疗对脊髓损伤家兔。采用Allen脊髓重物打击损伤法造模,并对家兔脊髓损伤后后肢行为功能评定。治疗结束后对比健康组、对照组和治疗组脊髓神经细胞凋亡情况,并检测治疗1 d、3 d、6 d、10 d后Bcl-2、Bax和caspase-3蛋白表达情况。结果 Basso分级评分结果显示,家兔脊髓损伤模型造模成功。造模后家兔脊髓神经细胞大量凋亡。与治疗前和对照组相比,给药富氢生理盐水后家兔神经细胞凋亡数明显减少。同时,对照组caspase-3蛋白表达高于健康组(P0.001)和治疗组(P0.001);与健康组相比,治疗组caspase-3蛋白表达水平较高(P0.01)。此外,治疗10 d后,治疗组Bax蛋白表达量低于对照组(t=2.263,P=0.035),但高于健康组(t=1.975,P=0.042);治疗组Bcl-2蛋白表达量高于对照组(t=0.463,P0.05),显著高于健康组(t=2.486,P=0.028);治疗组Bax/Bcl-2蛋白表达低于对照组,差异具有统计学意义(t=2.354,P=0.031)。结论腹腔注射富氢生理盐水显著降低了家兔脊髓损伤神经细胞凋亡及Bax/Bcl-2水平和caspase-3蛋白的表达。     

The prevalence of nonalcoholic fatty liver disease in Chinese adults screened by vibration controlled transient elastography and its diagnostic discrepancy compared with ultrasound

BackgroundThe prevalence of non-alcoholic fatty liver disease (NAFLD) in China as assessed using vibration-controlled transient elastography (VCTE) and its consistency with ultrasound is still unknown. We aimed to conduct a head-to-head comparison of consecutive measurements of NAFLD with ultrasound or VCTE to evaluate the discrepancy in the prevalence and distribution of NAFLD screened by two non-invasive techniques.MethodsWe collected VCTE and ultrasound examination data from 4,388 participants who underwent health check-ups at the Health Promotion Center of Jiangsu Province Hospital between January 2017 and December 2019. The major outcome was the presence of hepatic steatosis, which was defined as a median controlled attenuation parameter (CAP) ≥ 248 dB/m by VCTE or the definition of steatosis by ultrasound.ResultsAmong the 4,388 participants, 2,214 were diagnosed with NAFLD by VCTE (CAP ≥ 248 dB/m, 50.46%). Participants with severe steatosis (CAP ≥ 280 dB/m) were commonly male (77.94% vs. 50.38%, P < 0.001), were obese (45.09% vs. 1.79%, P < 0.001), had a worse metabolic profile, had elevated liver enzyme levels, and had advanced fibrosis. The prevalence of ultrasound-diagnosed NAFLD was 56.42%. After consistency analysis, VCTE and ultrasound showed moderate agreement regarding the diagnosis of NAFLD (κ = 0.475). We then compared the characteristics and clinical features of the four groups classified by the diagnosis results of the two techniques. NAFLD participants diagnosed by VCTE only were older, more obese, and had worse metabolic and biochemical profiles than NAFLD participants diagnosed by ultrasound only; in particular, the former had a higher proportion of abnormal alanine aminotransferase and aspartate aminotransferase levels and a higher proportion of advanced fibrosis than the latter.ConclusionsMore than half of Chinese adults were affected by NAFLD according to VCTE. Screening based on VCTE is more likely to identify NAFLD patients with severe clinical features than ultrasound. Therefore, VCTE is a more practical non-invasive tool for the screening and follow-up of NAFLD in China.     

重组葡激酶对急性坏死性胰腺炎大鼠心肌细胞凋亡调节基因表达的影响

目的观察SAP心肌功能损害时心肌细胞凋亡基因的表达及重组葡激酶对其的干预作用。方法63只SD大鼠随机分为假手术组、ANP6h、12h、18h组和葡激酶干预6h、12h、18h组（干预组），每组9只。采用5％的牛磺胆酸钠胰胆管逆行注射法建立ANP模型。观察心肌的病理变化，免疫组化法检测心肌Bel-2、Bax及N卜KB的表达。结果ANP组心肌片状坏死，NF-κB表达增加，为35．0～59．5；干预组心肌的病理变化减轻，NF-κB表达为10．5～17．0，较ANP组明显减少（P〈0．05）。ANP12h、18h组Bcl-2表达较假手术组显著减少（0．12～0．08υs0．31，P〈0．05），而Bax表达显著增加（1．48～2．83υs0．68）；干预12h、18h组Bcl-2表达较ANP组明显增加，达0．34～0．36（P〈O．05），而Bax表达明显减少，为1．06～1．10（P〈0．05）。结论ANP大鼠心肌NF-κB和Bax表达增加，Bcl-2表达减少；葡激酶干预后NF-κB及Bax的表达被抑制，Bcl-2的表达上调。     

脂氧素 A4预处理对糖尿病大鼠心肌缺血再灌注时Caspase －3、Bcl －2和 Bax 表达的影响

目的：观察脂氧素 A4预处理对链尿佐菌素诱导的糖尿病大鼠心肌缺血再灌注时 Caspase －3、Bcl －2和 Bax 表达的影响。方法雄性 SD 大鼠60只,随机分为 CS、CI／R、CLX、DS、DI／R、DLX 组共6组（n ＝10）。 CS、CI／R、CLX组采用腹腔注射链尿佐菌素55 mg／kg 制备糖尿病模型。 CI／R、DI／R组建立大鼠心肌缺血再灌注模型,结扎冠脉左前降支30 min,CS 组和 DS 组仅穿线不结扎,CI／R组和 DI／R组缺血前5 min 经股静脉注射2 mL／kg 生理盐水,CLX组和 DLX组缺血前5 min 经股静脉注射脂氧素 A42.5μg／kg。再灌注4 h 时处死大鼠,取心肌组织,采用 HE 染色法光镜下观察缺血心肌组织形态学改变,通过 RT －PCR 和 Western blot 法检测缺血心肌 Caspase －3、Bcl －2和 Bax的表达水平,并计算 Bcl －2与 Bax 表达的比值（Bcl －2／Bax）。结果CLX组和 DLX组心肌组织病理学损伤较 CI／R组和 DI／R组减轻,Caspase －3和 Bax 的表达量减少,Bcl －2的表达量增加,Bcl －2／Bax 比值升高（P ＜0.05）。结论脂氧素 A4预处理能够下调糖尿病大鼠心肌缺血再灌注时 Caspase －3和 Bax 的表达,上调 Bcl －2的表达,使 Bcl －2／Bax 比值升高。     

Effects of intracerebral hemorrhage and subsequent minimally invasive hematoma aspiration on expression of apoptosisrelated genes in rats

This study was to investigate the effects of intracerebral hemorrhage (ICH) and subsequent minimally invasive hematoma aspiration on the expression of apoptosis-related genes in rats. IV-collagenase was injected to the caudate nucleus of the rats to make ICH models. In the control group, 30 Sprague-Dawley (SD) rats were mock treated with saline instead of collagenase. Thirty SD rats with successful modeling were designated as the ICH group. Twenty-five SD rats with successful modeling and subsequent minimally invasive hematoma aspiration were designated as the therapy group. Expression of heat shock protein 70 (Hsp70), B-cell leukemia/lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) in the brain tissues was detected by immunohistochemical assays. The expression of Hsp70, Bcl-2 and Bax in the control group was very low, and significantly increased in the ICH group and the therapy group. At each indicated time point, Hsp70 expression in the therapy group was significantly lower than that of the ICH group, Bax expression in the therapy group was significantly lower than that of the ICH group and Bcl-2 expression in the therapy group was significantly higher than that of the ICH group. These results suggest that ICH led to increased expression of apoptosis-related genes in the brain tissues. Hematoma aspiration up-regulated ICH induced Bcl-2 expression while down-regulated ICH induced Hsp70 and Bax expression.     

半乳糖凝集素3表达抑制对人胃癌MGC-803细胞中Bcl-2和Bax表达的影响及其促凋亡作用

目的：探讨抑制半乳糖凝集素3（galectin-3）的表达对人胃癌MGC-803细胞凋亡的影响,为基因靶向治疗提供潜在靶点。方法：人胃癌MGC-803细胞分为siRNA干扰组、空白对照组和阴性对照组,siRNA干扰组MGC-803细胞转染靶向galectin-3的siRNA,空白对照组MGC-803细胞只加转染试剂,阴性对照组MGC-803细胞转染阴性对照的siRNA。采用流式细胞术和碘化丙啶（PI）染色法检测各组细胞凋亡率和细胞凋亡情况,Western blotting法检测各组细胞中galectin-3、Bcl-2和Bax蛋白表达情况。结果：与阴性对照组和空白对照组比较,siRNA干扰组细胞中galectin-3蛋白表达水平明显降低（P<0.05）。PI染色,siRNA干扰组凋亡细胞数量明显增加,凋亡细胞出现细胞核固缩、染色质浓集、新月形和凋亡小体等细胞凋亡特征,阴性对照组仅有少量散在的凋亡细胞。流式细胞术和Western blotting法检测,与阴性对照组和空白对照组比较,siRNA干扰组细胞凋亡率明显升高（P<0.05）,Bcl-2蛋白表达水平明显降低（P<0.05）,Bax蛋白表达水平未发生明显变化（P>0.05）。结论：抑制galectin-3表达可促进MGC-803细胞的凋亡,提示galectin-3对胃癌细胞发挥癌基因作用,具有成为靶向治疗靶点的可能性。     

臭牡丹提取物对H22荷瘤小鼠体内抑瘤作用及对相关蛋白表达的影响

目的 研究臭牡丹提取物对H22实体移植瘤的抑制作用及与B淋巴细胞瘤/白血病-2（Bcl-2）和Bcl-2相关X蛋白（Bax）表达的相关性。方法 皮下接种H22瘤株建立小鼠H22实体移植瘤模型。60只荷瘤鼠随机分为模型组、5-氟尿嘧啶（5-Fu）组和臭牡丹高、中、低剂量（2.18、1.09、0.55 g/kg）组,每组12只,各组分别给予生理盐水、5-Fu（25 mg/kg,1次/d）及不同浓度的臭牡丹提取物干预。连续给药14 d后处死,检测小鼠体质量变化、抑瘤率及脾脏和胸腺指数;Western blot法检测Bcl-2和Bax蛋白的表达水平。结果 与模型组比较,臭牡丹高、中剂量组小鼠的肿瘤质量均显著降低（P<0.01）,胸腺和脾脏指数显著增加（P<0.05）;同时Bax蛋白的表达水平显著上调,Bcl-2蛋白的表达显著降低（P<0.01）。结论 臭牡丹提取物能够抑制H22实体移植瘤生长,其机制可能与上调Bax/Bcl-2比值和免疫调节有关。     

量子点对口腔鳞癌Tca8113活细胞中bcl-2、bax的荧光标记观察

目的:探讨量子点对口腔鳞癌(OSCC)活细胞bcl-2、bax蛋白进行观察研究。方法:量子点QD605,QD545通过间接免疫荧光法,在激光共聚焦显微镜下观测人舌癌Tca8113活细胞内bcl-2、bax蛋白与量子点孵育0.5 h、1.0h、1.5 h和2.0 h的荧光成像。结果:量子点对OSCC活细胞中的bcl-2、bax蛋白的观察研究显示:在0.5 h～2.0 h时,QD605和QD545与bcl-2、bax蛋白结合,荧光从分布于胞浆边缘逐渐至胞浆。结论:量子点能对活细胞内蛋白进行观察。     

Melatonin protects against rotenone-induced cell injury via inhibition of Omi and Bax-mediated autophagy in Hela cells

Parkinson's disease is the second most common neurodegenerative disease, and environmental toxins such as rotenone play an important role in causing degeneration of dopaminergic neurons. Melatonin, a major secretory product of pineal, is recently reported to protect against rotenone-induced cell death in animal models. Yet, the mechanism involved in this protection needs to be elucidated. Here, we report that rotenone treatment (0-100 μM) decreased cell survival of Hela cells in a dose-dependent manner. At concentrations ranging from 0.1 to 100 μM, rotenone induced a dose-dependent increase in the expression of microtubule-associated protein 1 light chain 3 (LC3)-II, a protein associated with the autophagosomal membrane. Knockdown of Bax or Omi using shRNA inhibited 1 μM rotenone-induced autophagy. To determine whether melatonin would protect cells against rotenone-induced cell death and autophagy, we pretreated Hela cells with 250 μM melatonin for 24 hr in the presence of rotenone. Melatonin inhibited Bax expression and the release of the omi/HtrA2 into the cytoplasm induced by 1 μM rotenone. Melatonin 250 μM treatment also suppressed cell death induced by 0.1-100 μM rotenone and protected against the formation of LC3-II in cells exposed to 1 μM rotenone. This work demonstrates a novel role for melatonin as a neuroprotective agent against rotenone.